CA2067178C - Solid tumor treatment method and composition - Google Patents
Solid tumor treatment method and compositionInfo
- Publication number
- CA2067178C CA2067178C CA002067178A CA2067178A CA2067178C CA 2067178 C CA2067178 C CA 2067178C CA 002067178 A CA002067178 A CA 002067178A CA 2067178 A CA2067178 A CA 2067178A CA 2067178 C CA2067178 C CA 2067178C
- Authority
- CA
- Canada
- Prior art keywords
- tumor
- liposomes
- liposome
- peg
- lipid
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Expired - Lifetime
Links
Classifications
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K9/00—Medicinal preparations characterised by special physical form
- A61K9/10—Dispersions; Emulsions
- A61K9/127—Liposomes
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07F—ACYCLIC, CARBOCYCLIC OR HETEROCYCLIC COMPOUNDS CONTAINING ELEMENTS OTHER THAN CARBON, HYDROGEN, HALOGEN, OXYGEN, NITROGEN, SULFUR, SELENIUM OR TELLURIUM
- C07F9/00—Compounds containing elements of Groups 5 or 15 of the Periodic System
- C07F9/02—Phosphorus compounds
- C07F9/547—Heterocyclic compounds, e.g. containing phosphorus as a ring hetero atom
- C07F9/553—Heterocyclic compounds, e.g. containing phosphorus as a ring hetero atom having one nitrogen atom as the only ring hetero atom
- C07F9/5537—Heterocyclic compounds, e.g. containing phosphorus as a ring hetero atom having one nitrogen atom as the only ring hetero atom the heteroring containing the structure -C(=O)-N-C(=O)- (both carbon atoms belong to the heteroring)
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K47/00—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient
- A61K47/50—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates
- A61K47/69—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates the conjugate being characterised by physical or galenical forms, e.g. emulsion, particle, inclusion complex, stent or kit
- A61K47/6905—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates the conjugate being characterised by physical or galenical forms, e.g. emulsion, particle, inclusion complex, stent or kit the form being a colloid or an emulsion
- A61K47/6911—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates the conjugate being characterised by physical or galenical forms, e.g. emulsion, particle, inclusion complex, stent or kit the form being a colloid or an emulsion the form being a liposome
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K9/00—Medicinal preparations characterised by special physical form
- A61K9/10—Dispersions; Emulsions
- A61K9/127—Liposomes
- A61K9/1271—Non-conventional liposomes, e.g. PEGylated liposomes, liposomes coated with polymers
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P31/00—Antiinfectives, i.e. antibiotics, antiseptics, chemotherapeutics
- A61P31/04—Antibacterial agents
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P35/00—Antineoplastic agents
Abstract
A liposome composition for delivering a compound to a solid tumor via the bloodstream. The liposomes, which contain the agent in entrapped form, are composed of vesicle-forming lipids and between 1-20 mole percent of a vesicle-forming lipid deriva-tized with hydrophilic polymer, and have sizes in a selected size range between 0.07 and 0.12 microns. After intravenous adminis-tration, the liposomes are taken up by the tumor within 24-48 hours, for site-specific release of entrapped compound into the tu-mor. In one composition for use in treating a solid tumor, the compound is an anthracycline antibiotic drug which is entrapped in the liposomes at a concentration of greater than about 50 µg agent/µMole liposome lipid.
Description
WO 9INSS46 PCr/US9U/06211 ~ J~'~
.
.
SOLID TUMOR TREAT~3NT METHOD AND COMPOSITION
1. Fiela of the Invention The pre~ent invention relates to a liposome composi-tion and method, particularly for use in tumor diagnos-tics and/or theraF~Qtics.
.
.
SOLID TUMOR TREAT~3NT METHOD AND COMPOSITION
1. Fiela of the Invention The pre~ent invention relates to a liposome composi-tion and method, particularly for use in tumor diagnos-tics and/or theraF~Qtics.
2. References Allen, T.M., ~1~81) B~ ochem. Biophys . Acta 64û .
385397. Allen, T.M., and Ever~s~, J. (1983) J. Phar-macol. Exp. Therap. 226. 539-544.
-Altura, B.M. ~1980) Adv. Microc rc. 9 252-~94.
Alving, C.R. (1984) Biochem. Soc. Trans. 12.
342344.
Ashwell, G., and Morell, A.G. (1974) Adv. ~nzymo-logy 41, 99-128.
Czop, J.K. (1978) Proc. Natl. Acad. Sci. USA
75: 3831 .
Durocher, J.P., et al. ~1975) Blood 45:11.
Ellens, H., et al. (1981) Biochim. Biophys. Acta 674. 10-18.
Gabizon, A., Shiota, R. and Papahadjopoulcs, D.
(1989) J. Natl. Cancer Inst. 81, 1484-1488.
Gabizon, A., E~uberty, J., Straubinger, R.M;, Price, D. C. and Papahadjopoulos, D. (1988-1989) J. Liposome Re~h. 1, 123-135. ~
~ = ' WO 91~05546 PCr/US90/06211 _, 20`~`7 17$
Gregoriadis, G., and Ryman, B.E. (1972) Eur. J.
Biochem. 24, 485-491. =~
Gregoriadis, G., and Neerunjun, D. (lg74) Eur. J.
Biochem. 47, 179-185. =~
Gregoriadis, G., and Senior, J. (1980) F~BS Lett.
119, 43-46.
Greenberg, J.P., et al (1979) Blood 53:916.
Hakomori, S. 11981) Ann. Rev. Biochem. 50, 733-764.
Hong, K., Friend, D ., Glabe, C. and Papahad jopoulos (1984) Biochem. Biophys. Acta 732, 320-323 .
Hwan~- K.J., et al. (1980) Proc. Natl. Acad. Sci.
USA 77: 403d ~
Jain, K.~. '1989) J. Natl. Can. Inst. 81, 570-576.
Jonah, M.M., ~ al. (1975) Biochem. Biophys. Acta 401, 336-348.
Juliano, R.L., and ~:.amp, D. (1975) Biochem. ~io-phys. Res. Commun. 63. 651-659.
Karlsson, K.A. (198Z) In: Biological Membranes, Vol. 4, D. Chapman (ed.) Academic ~ress, .~.Y., pp. 1-74.
Kimelberg, H.K., et al . (1976) Cancer Res . 36, 2949-2 957 .
Kirby, C.J. and Gregoriadis (1984) Ir: Iiposome Technology, Vol. 3, G. Gregoriadis (ed. ) C RC Press, Boca Raton, FL., p. 19.
Lee, K.C., et al., J. Immunology 125:86 (1980).
10pez-8erestein, G., et al. (1984) Cancer Res. 4~, 375-378 .
Martin, F.J. (1990) In: Specialized Drug Delivery Systems - M~nllfactl~ring and Production Technology, P.
Tyle (ed. ) Marcel Dekker, New York, pp. 267-316.
Okada, N. ~1982) Nature 299:261.
Poste, G., et al., in "Liposome Technology" VoLume 3, page 1 (Gregoriadis, G., et~~al, eds . ), CRC Press, Boca Raton (1984);
WO 91~05546 PCI/US90/06211 .
20~7178 3 , ~ ~
Poznansky, M.J., and Juliano, R.L. ~1984) Pharmacol.
Rev. 36. 277-336.
Richardson, V.J., et aL. ~1979) Br. J. Cancer 40, 3543 .
5 Saba, T.~ 1970) Arch. In~ern. Med. 126.
1031-1052 .
Schaver, R. ~1982) Adv. Carbohydrate Chem. Biochem.
~0 131. ~ = ~
Scherphof, T., et al. ~1978) Biochim.Biophys. Acta 542, 2"6-307.
Senicr, J., and Gregoriadis, G. ~1982) FEBS Lett.
45, 109-114.
Senior, J., et al. (1985) Biochim. Biophys. Acta 839, 1-8.
Szoka, F., Jr., et al. (1978) Proc. Natl. Acad.
Sci . USA 75: 4194 .
Szoka, F., Jr., et al. (1980) Ann. Rev. Biophy~.
Bioeng . 9: 4 67 .
Weinstein, J.W., et al ., Pharn,a- The , 24: 207 ~1984).
Woodruff, J.J., et al. ~1969) J. Ex~. ~ed. 129:551.
3. Background of the Invention It would be desirable, for extravascular tumor 25 ~i ~gnnS~ c and therapy, to target an imaging or therapeu-tic c _ ~1 selectively to the tumor via the blood-stream. In diagnostics, such tar~eting could be used to provide a~ greater concentration of an imaging agent at the tumor site, as well as reduced background levels of 30 the agent in other parts of the body. Site-specific targeting would be useful in therapeutic treatment of tumors, to reduce toxic side effects and to increase the drug dose which can sa~ely be delivered to a tumor site.
WO 9t/05546 PCr/US90/06211 ?,~6~ 4 1iposomes have been~proposed as a drug carrier for intravenously (IV) adminlstered compounds, including both imaging and therapeutic compounds. However, the use of liposomes for site-specific targeting via the bloodstream 5 has been severely ~estricted by the rapid clearance of liposomes by cells of the reticuloendothelial system (RES). Typically, the RES will remove 80-95% of a dose of IY in~ected liposomes within one hour, effectively out-r:ompeting the selected target site for uptake of the lO liposomes.
A variety of factors which influence the rate of RES
uptake of liFosomes have been repQrted ~e.g., Gregoria-dis, 1974; Jon~h; Gregoriadis, 1972; Juliano; Allen, 1983; Kimelberg, 1976; Richardson; Lopez-Berestein;
Allen, 1981; Scherpho'; Gregoriadis, 1980; Hwang; Patel, 1983; Senior, 1985; Allen, 1983; Ellens; Senior, 1982;
Hwang; Ashwell; Hakomori; K~rlsson; Schauer; Durocher;
Greenberg; Woodruff; Czop; and C~da). Briefly, liposome size, charge, degree of lipid saturat~ ~ n, and surface moieties have all been implicated ln liposom~ ~learance by the RES. However, no single factor id~ntified to date has been effective to provide lonq blood halflife, and more particularly, a relatively high percentage of lipo-somes in the bloodstream 24 hours after in~ection.
In addition to a long blood halflife, effective drug delivery to a tumor site would also require that the liposomes be capable of penetrating the continuous enfio-thelial cell layer ana underlying basement membrane surrounding the vessels supplying blood to a tumor.
Although tumors may present a damaged~ leaky endothelium, it has generally been recognized that for liposomes to reach tumor cells in effectiYe amounts, ~ the liposomes would have to possess me~ h~n~ emC whlch facilitate their passage through the endothelial cell barriers and adja-~ ~, s; .
_ WO 91/05546 PCr/US90/06211 2067i78 cent basement membranes, particularly in view of the low blQod flow to tumors and hence limited exposure to circu-lating liposomes ~Weinstein). Higher than normal inter-stitlal pressures found within most tumors would also 5 tend to reduce the opportunity for extravasation of lipo-somes by creating a an outward transvascular movement of fluid from the tumor (Jain). As has been pointed out, it would be unlikely to design a liposome which would over-come these barriers to extravasation in tumors and, at 10 the sa~ time, evade RES recognltion and uptake (~oz-nanski ) .
In fact, ~tudies reported to date indicate that even where the permea~ility of blood vessels increases, extra-vasation of conven~ional liposomes through the vessels 15 does not increase sigr.ificantly (Poste). Based on these findings, it was concluded that although extravasation of liposomes from capillaries compromised by disease may be occurring on a limited scale beiow detection levels, its therapeutic potential would be minlmal ~L ~ste) .
385397. Allen, T.M., and Ever~s~, J. (1983) J. Phar-macol. Exp. Therap. 226. 539-544.
-Altura, B.M. ~1980) Adv. Microc rc. 9 252-~94.
Alving, C.R. (1984) Biochem. Soc. Trans. 12.
342344.
Ashwell, G., and Morell, A.G. (1974) Adv. ~nzymo-logy 41, 99-128.
Czop, J.K. (1978) Proc. Natl. Acad. Sci. USA
75: 3831 .
Durocher, J.P., et al. ~1975) Blood 45:11.
Ellens, H., et al. (1981) Biochim. Biophys. Acta 674. 10-18.
Gabizon, A., Shiota, R. and Papahadjopoulcs, D.
(1989) J. Natl. Cancer Inst. 81, 1484-1488.
Gabizon, A., E~uberty, J., Straubinger, R.M;, Price, D. C. and Papahadjopoulos, D. (1988-1989) J. Liposome Re~h. 1, 123-135. ~
~ = ' WO 91~05546 PCr/US90/06211 _, 20`~`7 17$
Gregoriadis, G., and Ryman, B.E. (1972) Eur. J.
Biochem. 24, 485-491. =~
Gregoriadis, G., and Neerunjun, D. (lg74) Eur. J.
Biochem. 47, 179-185. =~
Gregoriadis, G., and Senior, J. (1980) F~BS Lett.
119, 43-46.
Greenberg, J.P., et al (1979) Blood 53:916.
Hakomori, S. 11981) Ann. Rev. Biochem. 50, 733-764.
Hong, K., Friend, D ., Glabe, C. and Papahad jopoulos (1984) Biochem. Biophys. Acta 732, 320-323 .
Hwan~- K.J., et al. (1980) Proc. Natl. Acad. Sci.
USA 77: 403d ~
Jain, K.~. '1989) J. Natl. Can. Inst. 81, 570-576.
Jonah, M.M., ~ al. (1975) Biochem. Biophys. Acta 401, 336-348.
Juliano, R.L., and ~:.amp, D. (1975) Biochem. ~io-phys. Res. Commun. 63. 651-659.
Karlsson, K.A. (198Z) In: Biological Membranes, Vol. 4, D. Chapman (ed.) Academic ~ress, .~.Y., pp. 1-74.
Kimelberg, H.K., et al . (1976) Cancer Res . 36, 2949-2 957 .
Kirby, C.J. and Gregoriadis (1984) Ir: Iiposome Technology, Vol. 3, G. Gregoriadis (ed. ) C RC Press, Boca Raton, FL., p. 19.
Lee, K.C., et al., J. Immunology 125:86 (1980).
10pez-8erestein, G., et al. (1984) Cancer Res. 4~, 375-378 .
Martin, F.J. (1990) In: Specialized Drug Delivery Systems - M~nllfactl~ring and Production Technology, P.
Tyle (ed. ) Marcel Dekker, New York, pp. 267-316.
Okada, N. ~1982) Nature 299:261.
Poste, G., et al., in "Liposome Technology" VoLume 3, page 1 (Gregoriadis, G., et~~al, eds . ), CRC Press, Boca Raton (1984);
WO 91~05546 PCI/US90/06211 .
20~7178 3 , ~ ~
Poznansky, M.J., and Juliano, R.L. ~1984) Pharmacol.
Rev. 36. 277-336.
Richardson, V.J., et aL. ~1979) Br. J. Cancer 40, 3543 .
5 Saba, T.~ 1970) Arch. In~ern. Med. 126.
1031-1052 .
Schaver, R. ~1982) Adv. Carbohydrate Chem. Biochem.
~0 131. ~ = ~
Scherphof, T., et al. ~1978) Biochim.Biophys. Acta 542, 2"6-307.
Senicr, J., and Gregoriadis, G. ~1982) FEBS Lett.
45, 109-114.
Senior, J., et al. (1985) Biochim. Biophys. Acta 839, 1-8.
Szoka, F., Jr., et al. (1978) Proc. Natl. Acad.
Sci . USA 75: 4194 .
Szoka, F., Jr., et al. (1980) Ann. Rev. Biophy~.
Bioeng . 9: 4 67 .
Weinstein, J.W., et al ., Pharn,a- The , 24: 207 ~1984).
Woodruff, J.J., et al. ~1969) J. Ex~. ~ed. 129:551.
3. Background of the Invention It would be desirable, for extravascular tumor 25 ~i ~gnnS~ c and therapy, to target an imaging or therapeu-tic c _ ~1 selectively to the tumor via the blood-stream. In diagnostics, such tar~eting could be used to provide a~ greater concentration of an imaging agent at the tumor site, as well as reduced background levels of 30 the agent in other parts of the body. Site-specific targeting would be useful in therapeutic treatment of tumors, to reduce toxic side effects and to increase the drug dose which can sa~ely be delivered to a tumor site.
WO 9t/05546 PCr/US90/06211 ?,~6~ 4 1iposomes have been~proposed as a drug carrier for intravenously (IV) adminlstered compounds, including both imaging and therapeutic compounds. However, the use of liposomes for site-specific targeting via the bloodstream 5 has been severely ~estricted by the rapid clearance of liposomes by cells of the reticuloendothelial system (RES). Typically, the RES will remove 80-95% of a dose of IY in~ected liposomes within one hour, effectively out-r:ompeting the selected target site for uptake of the lO liposomes.
A variety of factors which influence the rate of RES
uptake of liFosomes have been repQrted ~e.g., Gregoria-dis, 1974; Jon~h; Gregoriadis, 1972; Juliano; Allen, 1983; Kimelberg, 1976; Richardson; Lopez-Berestein;
Allen, 1981; Scherpho'; Gregoriadis, 1980; Hwang; Patel, 1983; Senior, 1985; Allen, 1983; Ellens; Senior, 1982;
Hwang; Ashwell; Hakomori; K~rlsson; Schauer; Durocher;
Greenberg; Woodruff; Czop; and C~da). Briefly, liposome size, charge, degree of lipid saturat~ ~ n, and surface moieties have all been implicated ln liposom~ ~learance by the RES. However, no single factor id~ntified to date has been effective to provide lonq blood halflife, and more particularly, a relatively high percentage of lipo-somes in the bloodstream 24 hours after in~ection.
In addition to a long blood halflife, effective drug delivery to a tumor site would also require that the liposomes be capable of penetrating the continuous enfio-thelial cell layer ana underlying basement membrane surrounding the vessels supplying blood to a tumor.
Although tumors may present a damaged~ leaky endothelium, it has generally been recognized that for liposomes to reach tumor cells in effectiYe amounts, ~ the liposomes would have to possess me~ h~n~ emC whlch facilitate their passage through the endothelial cell barriers and adja-~ ~, s; .
_ WO 91/05546 PCr/US90/06211 2067i78 cent basement membranes, particularly in view of the low blQod flow to tumors and hence limited exposure to circu-lating liposomes ~Weinstein). Higher than normal inter-stitlal pressures found within most tumors would also 5 tend to reduce the opportunity for extravasation of lipo-somes by creating a an outward transvascular movement of fluid from the tumor (Jain). As has been pointed out, it would be unlikely to design a liposome which would over-come these barriers to extravasation in tumors and, at 10 the sa~ time, evade RES recognltion and uptake (~oz-nanski ) .
In fact, ~tudies reported to date indicate that even where the permea~ility of blood vessels increases, extra-vasation of conven~ional liposomes through the vessels 15 does not increase sigr.ificantly (Poste). Based on these findings, it was concluded that although extravasation of liposomes from capillaries compromised by disease may be occurring on a limited scale beiow detection levels, its therapeutic potential would be minlmal ~L ~ste) .
4. Summary of the Invention One general ob~ect of the invention is to provide a liposome composition and method which ix ef ~ective or tumor targeting, for lot ~1 i 7~ n~ an imaging or anti-tumor 25 agent selectively at therapeutic dose levels in systemic, extravascular tumors.
The lnvention includes, in one aspect, a liposome composition for use in localizing a compound in a solid tumor, as defined in Section IV below, via the blood-30 stream comprising: The liposomes forming the composition(i) are composed of vesicle-forming lipids and between l-20 mole percent of an vesicle-forming lipid derivatized with a hydrophilic polymer, and (ii) have an average size in a selected size range between about 0 . 07-0 .12 microns .
-The compound i5 con-ained in the liposomes in en,-apped form li.e., associated with the liposome membrane o~
encapsulated within the internal aqueous compartment of the liposome). In this context, vesicle-forming lipid is defiAed as any lipid that by itself or in comb nation with other lipids forms bilayer structures.
In a preferred embodiment, the hydrophilic polymer is polyethyleneglycol or poly lactic poly glycolic acid havir.~ a molecular weight between about 1, 000-5, OQ0 daltons, and is derivatized to a phospholipid.
For u~e in tumor treatment, the compound in one embodiment is im anthracycline antibiotic or plant alka--loid, at least about 80% of the ~ ?ou.,~ is in liposome-entrapped form, and the drug is present in the liposomes at a concentration of a~ least about 20 llg compound/umole liposome lipid in the case of the anthracycline antibio-tics and and 1 ug/umoles lipi:l in the case of the plant alkaloids .
In a related aspect, the invention inclu~os a com-position of liposomes characterized by:
(a) liposomes composed of vesicle-forming lipids and between 1-20 mole percent of an vesicle-fo~ina lipid derivatized with a hydrophilic polymer, ~b) a blood lifetime, as measured by the percent of a liposomal marker present in the blood 24 hours after IV
administration which is several times greater than that of liposomes in the absence of the derivatized lipids;
(c) an average liposome size in a selected size range between about 0.07-0.12 microns, and (d) the compound in liposome-entrapped form.
Also disclosed is a method of preparing an agent for loc~l;7?(tion in a solid tumor, when the agent is adminis-tered by IV injection. In this case, following IV ad~.i-nistration the agent is carried through the bloodstream A
WO 91/05546 PCr/US90/062tl 20~71~8 in liposome-entrapped form with little leakage of the drug durlng the first 48 hours post injection. By virtue of the low rate of RES uptake during this period, the liposomes have the opportunity to distribute to and enter 5 the tumor. Once within the interstitial spaces of the tumor, it is not necessary that the tumor cells actually internalize the liposomes. The entrapped agent is re-leased from the liposome in close proximity to the tumor calls over a period of days to weeks and is free to 10 further Penetrate into the tumor mass ~by a process of diffusion) and enter tumor cells directly - exerting its anti-proliferative activity. The method includes entrap-ping the agent in liposomes of the type characterized above. One liposome composition preferred for transpo=t-15 ing anthracycllne antibiotic or plant alkaloid anti-tumor agents to systemic solid ~ umors would contain high phase transition phospholipids and ~holesterol as this type of liposome does not tend to rel~ase these drugs while circ~ t ~ n~ through the bloodstream du i ~lg the f_rst 24-20 48 hours following administration.
In another aspect, the invention incl-~ldes a method for localizing a ~~ ~ou..d in a solid tumor ir a subject.
The method includes preparing a composition of liposo.nes ~i) composed of vesicle-forming lipids and between 1-20 25 mole percent of an vesicle-forming lipid derivatized with a hydrophilic polymer, (ii) having an average siz~ in a selected size range between about 0 . 07-0 .12 microns, and ~iii) c~nt~n~n~ the compound in liposome-entrapped form.
The compositi on is in jected IV in the sub~ect in an 30 amount sufficient to localize a therapeutically effective dose of the agent in the solid tumor.
These and other objects and features of the present invention will become more fully apparent when the fol-lowing detailed description of the invention is read in .
: -conjunc' ion with the accompanying drawings.
Brief Description of the Drawings Figure 1 illustrates a general reaction scheme for 5 derivatizing a vesicle-forming lipid amine with a polyal-kylether;
Figure 2 is a reaction scheme for preparing phospha-tidyleth~nolamine (PE) derivatized with polyethylene-qlycol via a cyanuric chloride linking agent;
Figure 3 illu$trates a reaction scheme for preparing phosphatidylethanolamine (PE) derivatized with polyethy-leneglycol by ~eans of a d;;mi~A~ole activating reagen~;
Figure 4 illustrates a reaction scheme for preparing phos~h~tidylethanolamine (PE) derivatized with polyethy-leneglycol by means of a trifluo,. ~hAn~o sulfonate reagent;
Figure 5 illustrates a v~sicle-forming lipid deriva-tized with polyethyleneglycol ~hrough a peptide (A), ester (8), and disulfide (C) linkag~;
Figure 6 illustrates a reaction sci;eme so- p-eparing phosphatidyle1-hAnol Am; n.o (PE) derivatiz~d with poly lactic acid or polyglycolic acid;
Figure 7 is a plot of liposome re.sidenc~ times in the blood, expressed in terms of percent injected dose as a function of hours after IV injection, for PEG-PE lipo-somes containing different amounts of ~hos~hAtidylglyce-rol;
Figure 8 is a plot slmilar to that of Figure 7, showing blood res; d~nce times of liposomes composed of pred: inAntly unsaturated phospholipid components;
Figure 9 is a plot similar to that of Figure 7, showing the blood residence times of PEG-coated liposomes (solid triangles) and conventional, uncoated liposomes ~ solid circles );
A
Figure 10 i8 a plot showing the kinetics of ., doxorubicin clearance from the blood of beagle dogs, for -drug administered IV in free form (open circles), in liposomes formulated with saturated phospholipids and llydLoy~llated phosphatidylinositol (HPI) (open squares), and in liposomes coated with PEG (open triangles);
Fiqures lL~ and llB are plots of the time course of doxorubicin uptake from the bloodstream by heart (solid ~l;i '-), muscle (solid circles), and tumor (solid triangles) for drug administered IV in free llA and PEG-1 ;ro~ 1 (llB) form;
Figure 12 is a plot of the time course of uptake of doxorubicin from the bluod~L~al,l by J-6456 tumor cells implanted interperitoneally (IP) in mice, as measured as total drug (filled ,i;; '-) as drug associated with tumor cells (solid circles) and liposome-associated form (solid triangles);
Figures 13A-13D are light mi~Loyl~plls showing localization of liposomes (small dark stained particles) in Kupfer cells in normal liver (13A), in the interstitial fluid of a C-26 colon carcinoma implanted in liver in the region of a capillary supplying the tumor cells (13B) and in the region of actively dividing C-26 tumor cells implanted in liver (13C) or subcutaneously (13D);
Figures 14A-14C are plots showing tumor size growth in days following subcutaneou6 implantation of a C-26 colon carcinoma, for mice treated with a saline control (open circles), doxorubicin at 6 mg/kg (filled circles), epirubicin at 6 mg/kg (open triangles), or PEG-liposome entrapped epirubicin at two do6es, 6 mg/kg (filled triangles) or 12 mg/kg (open squares) on days 1, 8 and 15 (14A); for mice treated with saline (solid line), 6 mg/kg epirubicin (closed circles), 6 mg/kg epirubicin plus empty liposomes, (open circles), or PEG liposome -9a- 206il 78 entrapped at two doses, 6 mg/kg (filled triangles) and 9 mg/kg topen squares) on days 3 and 10 (14B) or days 10 and 17 ( 14C);
Figure 15 i8 a plot 6howing percent survivors, in 5 days following interperitoneal implantation of a J-6456 lymphoma, for animals treated with doxorubicin in free form (c$osed circles) or PEG-liposomal form (solid triangles), or untreated animals (filled squares); and Figure 16 is a plot similar to that in Figure 14, 10 showing tumor size growth, in days following subcutaneous implantation of a C-26 colon carcinoma, for animals treated with a saline control (solid line), or animals treated with 10 mg/kg doxorubicin in free form (filled circles), or in conventional liposome~; (filled 15 triangles).
lO 2067 1 7~
Detailed Description of the Il~ven ion I. PreparatiOn of Derivatized Lipids Figure 1 shows a general reaction scheme f or prepa-5 ring a vesicle-forming lipid derivatized a biocompatible, hydrophilic polymer, as exemplified by polyethylene glycol (PEG), polylactic acid, and polyglycolic acid, all of which are readily water soluble, can be coupled to vesicle-forming lipids, and are tolerated in vivo without 10 toxic effects. The hydrophilic polymer which is em-ployed, e.g., PEG, is preferably capped by a methoxy,ethoxy or other unreactive group at one end or, alte-na-tively, has a chemical group that is more highly rea~~ive at one end than the other. The polymer is activated at A
WO 91/0~46 PCr/US90/06211 .
2~67178 one of its ends by reaction with a suitable activating agent, such as cyanuric acid, diimadozle, anhydride reagent, or the like, as described below. The activated compound is then reacted with a vesicle-~orming lipid, 5 such as a diacyl glycerol, including diacyl phosphogly-cerols, where the two hydrocarbon chains are typically between 14-22 carbon atoms in length and have Yarying degrees of saturation, to produce the derivatized lipid.
phosrll~tidylethanol-amine (PE) is an example of a phos-10 pholipid which is preferred for t~Lis purpose since itcontains a reactive amino group which is convenient for coupling to the activated polymers. Alternatively, the lipid group may be activated for reaction with the poly-mer, or the two groups may be ~oined in a concerted 15 coupling reaCtiOn, according to known coupling methods.
PEG capped at one end with a methoxy or ethoxy group can be obtained commercially in a variety of polymer sizes, e.g., 500-20,000 dalton molecular weights.
The vesicle-forming lipid is pr~rQrably one having 20 two hydrocarbon chains, typically acyl chai~s, and a polar head group. Included in this class are the phos-pholipids, such as rh~srh~tidylcholine (PC), PE, phos-rh~ acid (PA), phosphatidylinositol (PI), and sphin-gomyelin (SM), where the two hydrocarbon chains are 25 typically between about 14-22 carbon atoms in length, and have varying degrees of unsaturation. Also included in this class are the glycolipids, such as cerebrosides and g~n~l i os1 ~l~os .
Another vesicle-forming lipid which may be employed 30 is cholesterol and related sterols. In general, choles-terol may be less tightly anchored to a lipid bilayer membrane, particularly when derivatized with a high molecular weight polymers, such as polyalkylether, and therefore be less effectlve in promoting liDosome evasion 6 PCr/US90/06211 of the RES in the bloodstream.
More generally, and as defined herein, "vesicle-forming lipid" is intended to include any amphipathic lipid having hydrophobic and polar head group moieties, 5 and which (a) by itself can form spontaneously into bilayer vesicles in water, as exemplified by phospholi-pids, or (~) is stably incorporated into lipid bilayers in combination with phospholipids, with its hydrophobic moiet~ in contact with the interior, hydrophobic region 10 of the b_layer membrane, and its polar head group moiety oriented toward the e~terior, polar surface of the mem-brane. An example of a latter type of vesicle-forming lipid is cholesterol and cholesterol derivatives, such as cholesterol sulfate and cholesterol hemisuccinate.
According to one important feature of the invention, the vesicle-forming lipid may be a relatively fluid lipid, typically meaning tha' the lipid phase has a relatively low liquid to liq.~id-crystalline melting temperature, e.g., at or below room temperature, or 20 relatively rigid lipid, meaning tha. the lipid has a relatively high melting temperature, ~.g., up to 60C.
As a rule, the more rigid, i.e., saturated lipiîl.~, con-tribute to greater membrane rigidity in a lipid bilayer structure and also contribute to greater bilayer stabi-25 lity in serum. Other lipid components, such as choleste-rol, are also known to contribute to membrane rigidlty and stability in lipid bilayer structures. A long ch~in (e.g. C-18) saturated lipid plus cholesterol is one preferred composition for delivering anthracycline anti-30 biotic and plant alkaloids anti-tumor agents to solid tumors since these liposomes do not tend to release the drugs into the plasma as they circulate through the bloodstream and enter the tumor during the first 48 hours following injection. Phospholipids whose acyl chains WO 91/05546 PCr/US90/06211 .
13 ~
have a variety of degrees of saturation can be obtained commercially, or prepared according to puolished methods.
Figure 2 shows a reaction scheme ~or producing a PE-PEG lipid in which the PEG is derivatized to PE through a 5 cyanuric chloride group. Details of the reaction are provided in Example 1. ~riefly, methoxy-capped PEG is activated with~ cyanuric chloride ln the presence in sodium carbonate under conditions which produced the activated PEG compound shown in the figure. This mate-10 rial is ~urified to remove unreacted cyanuric acid. Theactivated PE5 compound is reacted with PE in the presence of triethyl amine to produce the desired PE-PEG compound shown in the figure. The yield is about 8-10% with respect to initial riuantities of PEG.
The method just described may be applied to a vari-ety of lipld amines, lnr11-Aing PE, cholesteryl amine, and glycolipids with sugar-amine g oups.
A second method of coupling a polyalkylether, such as capped PEG to a lipid amine is ' llustrated i~. Figure 20 3. Here the capped PEG is activated witn a ~arbonyl m~ 7Ole coupling reagent, to form ~he activated imidazole compound shown in Figure 3. RePction with a lipid amine, such as PE leads to PEG coupli~g to the lipid through an amide linkage, as illustrated in the 25 PEG-PE compound shown in the figure. Details of the reaction are given in Example 2.
A third reaction method for coupling a capped poly-alkylether to a lipid amine is shown in Figure 4. Here PEG is ~irst protected at its OH end by a trimethylsilane 30 group. The end-protection reaction is shown in the figure, and involves the reaction of trimethylsilylchlo-ride with PEG in the presence of triethylamine. The protected PEG is then reacted with the anhydride of trifluoromethyl sulfonate to form the PEG compound acti-= =, , .
WO 91/05546 PCr/US90/06211 =-G~ 14 vated with trifluoromethyl sulfonate. Reaction of the activated compound wlth a lipid amine, such as PE, in the presence of = trie~hylamine, gives the desired derivatized lipid product, such as the PEG-PE compound, in which the 5 lipid amine group is coupled to the polyether through the terminal methylene carbon in the polyether polymer. The trimethylsilyl protective group can be released by acid treatment, as indicated in the figure, or, alternatively, by reaction with a quaternary amine fluoride salt, such l0 as the fluoride salt of tetrabutylamine.
It will be appreciated that a variety of known coupling reactions, in addition to those ~ust described, are suitable for preparing vesicle-forming lipids deriva-tized with hydrophilic polymers such as PEG,. For e~am-15 ple, the sulfonate anhyd. ide coupling reagent illustratedin Figure 4 can be used to ~oin an activated polyalkyl-ether to the hydroxyl group of an amphipathic lipid, such as the 5'-OH of cholesterol. Other reactive lipid groups, such as an acid or ester lipid group may also be 20 used for coupling, according to known coupling methods.
For example, the acid group of phosphatidi- acid can be activated to form an active lipid anhydride, by reaction with a suitable anhydride, such as acetic ~nhydride, a~d the reactive lipid can then be ~oined to a protected 25 polyalkylamine by reaction in the presence of an isothio-cyanate reagent. - -In another embodiment, the derivatized lipid c~m-ponents are prepared to include a labile lipid-polymer linkage, such as a peptide, ester, or disulfide linkage, 30 which can be cleaved under selective physiological condi-tions, such as in the presence of peptidase or esterase enzymes or reducing agents such as glutathione present in the bloodstream. Figure 5 shows exemplary lipids which are linked through (A) peptide, (B), ester, and (C), disul'ide containing linkages. The pep~ide-linked com-pound can be prepared, for example, by first coupling ~a polyalkylether with the N-terminal amine of the t-i~ep-tide shown, e.g., via the reaction shown in Figure 3.
5 The peptide carboxyl g-oup can then be coupled to a lipid amine group through a carbodiimide coupling reAgent con-ventionat ly . The ester linked compound can be prep2red, for example, by coupling a lipid acid, such as phospha~i-dic ac~ d, to the terminal alcohol group of a polyalkyl-10 ether, u~ing alcohol via an anhydride coupling agent.Alternatively, a ~hort linkage fragment c~nt~;n~rlg an internal ester bond and suitable end groups, such as primary amine groups can be used to couple the polyalkyl-ether to the amphipathic lipid through amide or ca:bamate 15 linkages. Similarly, the linkage fragment may contain an internal disulfide linkage, for use in forming the com-pound shown at C in Figure 5. Polymers coupled to phos-pholipids via such reversible inkages are useful to provide high blood levels of liposom~s which cont~in them 20 for the first few hours post injection. After this period, plasma components cleave the :ev~rsible bonds releasing the polymers and the "unprotected" i ~osomes are rapidly taken up by the RES.
Figure 6 illustrates a method for derivatizlng 25 polylactic acid with PE. The polylactic acid is reacted, in the presence of PE, with dicyclohexylcarboimide (DCCI), as detailed in Example 4. Similarly, a vesicle-forming lipid derivatized with polyglycolic acid may be formed by reaction of polyglycolic acid or glycolic acid 30 with PE in the presence of a suitable coupling agent, such as DCCI, also as detailed in Example 4. The vesi-cle-forming lipids derivatized with either polylac.ic acid or polyglycolic acid form part of the inven~ion herein. Also forming part of the inventiOn are liposomes A
WO 91/05546 PCr/US90/06211 1~ ~
?,~6 containing these derlvatized Lipids, in a 1-20 moLe percent .
II. Preparation of Liposome Composition 5 A. ~ipid Components The lipid components used in forming the liposomes of the invention may be selected from a variety of vesi-cle-forming lipids, typically including phospholipids, sphing~lipids and sterols. As will be seen, one require-lO ment of the liposomes of the present invention is longblood circulation lifetime. It is therefore useful to establish a standardized measure of blood lifetime which can be used for evaluating the effect =of lipid components on blood halflife.
one method used for evAlu~t;n~ l;ros~ - circulation tlme in vivo measures the distribution of IV injected liposomes in t~e bloodstream and the primary organs of the RES at selected times after injection. In the stan-dardized model which is used herein, RE~S uptake is mea-20 sured by the ratio of total liposomes l~ the bloodstream to total liposomes in the liver and spleen, the principal organs of the RES. In practice, age and sex matched mice are in~ected IV through the tail vein with a radiolab~' sd liposome composition, and each time point i~ determined 25 by measuring total blood and combined liver and spleen radiolabel counts, as detailed in Example 5.
Since the liver and spleen account for nearly 100%
of the initial uptake of liposomes by the RES, the blood-/RES ratio ~ust described provides a good approximation 30 of the extent of uptake from the blood to the RE~ ln vivo. For example, a ratio of about l or greater indi-cates a pr~ m; nAn~`p of injected liposomes remaining in the bloodstream, and a ratio below about l, a predomi-nance of Iiposomes in the RES. For most of the lipid WO 91/05546 PCr/US90/06211 compositions of interest, blood/RES ratios were calcu-lated at 1,2, 3, 4, and 24 hours post injection.
The liposomes of the present invention include 1-20 mole percent of the vesicle-forming lipid derivatized 5 with a hydrophilic polymer, described in Section I.
According to one aspect of the invention, it has been discovered that blood circulation halflives in these liposomes is largely independent of the degree of satura-tion of the phospholipid components making up the lipo-l0 somes. That is, the phospholipid components may becomposed of pr,orl~ ;n~ntly of fluidic, relatively unsatu-rated, acyl chains, or of more saturated, rigidifying acyl chain components. This feature of the invention is seen in Example 6, which Pl~m~ nPC blood/RES ratios in 15 liposomes formed with PE~-PE, cholesterol, and PC having varying degrees of saturation (Table 4)~. As seen from the data in Table 5 in the example, high blood/RES ratios were achieved in subst~nt;~1 1y ail of the liposome for-m111 at ~ t~nCi, t n~lprpn~pnt of the extenL of lipid ur,satura-20 tion in the bulk PC phospholipid, an~ no systematictrend, as a function of degree of lipid SaLuratiOn, waS
observed .
Accordingly, the vesicle-forming lipids may ~e selected to achieve a selected degree of fluidity or 25 rigidity, to control the stability of the liposomes in serum and the rate of release of entrapped drug from the liposomes in the bloodstream andtor tumor. The vesicle-forming lipids may also be selected, in lipid saturation characteristiCs, to achieve desired liposome preparation 30 properties. It is generally the case, for example, that more fluidic lipids are easier to formulate and down-size by extrusion and homogenization methods than more rigid lipid compositions.
:
, ~==
. '8 20671 78 Similarly, it h2s been found that the percen_age o~
cholesterol in the liposomes may be va_ied over a wi~e range wi~chout si~nifican~ effect on observed blood~REs ratios. The studies presenced in Example 7A, with refer-ence to Table 6 therein, show virtually no change in blood/RES ratios in the range of cholesterol between 0-30 mole percent.
~ It has also been found, in studies conducted in support o~ the invention, that blood~RES ratios are also relatively unaffected by the presence of charged lipid components, such as phos~hatidylglycerol tPG). This can be seen from Figure 7, which plots percent loss of encap-sulated marker for PEG-PE liposomes cont~in;ns either 4.7 mole percent PG ~triangles) or 14 mole percent P~; (cir-cles). Virtually no difference in liposome retention in the bloodstream over a 24 hour period was observed. The option of including negative charge in the liposome without aggravating RES uptake provides a number o~
potential advantages. Liposomes su~pensions which con-tain negative charge tend to be less s~nsi.ive to aggre-gation in high ionic strength buffers and hence physical stability is enhanced. Also, negative cha~go p~i~sent in the liposome membrane can be used as a formulat on ~ol to effectively bind high amounts of cationic drugs.
The vesicle-forming lipid derivatized wit~ a hydro-philic polymer is present in an amount preferably between about 1-20 mole percent, on the basis of moles of deriva-tized lipid as 2 percentage of total moles of vesicle-forming lipids. It will be appreciated that a lower mole ratio, such as less than 1. O mole percent, may be d}J~L~Liate for a lipid derivative with a large molecular weight polymer, such as one havlng a molecular weight o~ lOO kilodaltons. As noted in Section I, the hydrophilic polymer in the derivatized lipid preferably has a molecular weight WO 91/OSS46 - - PCr/l~S90/06211 ~ 20~7178 between about 200-20, 000 daltons, and more pre~erably between about 500-5, 000 daitons. Example ~B, which Am; nPC the effect of very short ethoxy ether moieties on blood/RES ratLos indlcates that polyether moLeties of 5 greater than about 5 carbon ethers are required to achieve signiflcant PnhAnc~=mPn~ of blood/RES ratlos.
B. Preparing the Liposome Composltion The liposomes may be prepared by a variety of tech-10 nlques, such as those detalled in Szoka et al, 1980. Onemethod for preparlng drug-containlng liposomes is the reYerse phase e~-aporation method described by Szoka et al and ln U.S. Patent No. 4,235,871. The reverse phase evaporatlon veslcles (REVs) have typical average slzes 15 between about 2-4 microl~iC and are preri( ~ nAntly oligo-l: ~ ~ l 1 Ar~ that 15~ coDtain one or a few lipid bilayer shells. The method is detaile~ in Example 4A.
~ lUlt~ 11 Ar vesicles (~V9) can be formed by simple lipid-film hydration techniques. In this proce-20 dure, a mixture of liposome-forming lipids of t he type detailed above dissolved in a suitable organ~ c solvent is evaporated in a vessel to form a thin fllm, whlch ls then covered by an aqueous medium, as detalled ln Example AB.
The lipid film hydrates to form ~Vs, typlcally with 25 sizes between about 0.1 to 10 microns.
In accordance with one important aspect of the invention, the liposomes are prepared to have suhstan-tially h~ ouS sizes in a selected slze range between about 0 . 07 and 0 .12 mlcrons . In partlcular, lt has been 30 discovered that liposomes in this size range are r~adily able to extravasate into solid tumors, as discussed in Sectlon III below, and at the same tlme, are capable of carrying a substantlal drug load to a tumor (unlike small ~ li -llAr vesicles, whlch are severely restricted in WO 91/05546 PCr/US90/06211 drug-loading capaclty). ~
One effecti~ve sizing method for ÆVs and MLVs in-volves extruding an aqueQus suspension of the liposomes through a series o~ = polycarbonate membranes having a selected uniform pore si2e in the range of 0. 03 to 0 .2 micron, typically 0.05, 0.08, 0.l, or 0.2 microns. The pore size of the membrane corresponds roughly to the largest sizes of liposomes produced by extrusion through that membrane, particularly where the preparation is l 0 extruded two or more times through the same membrane .
This method of liposome sizing is used in preparing homogeneous-si2e REV and ~qLV compositions described in the examples below. A more recent method involves extru-sion through an asymmetric ceramic i~ilter. The method is detalled in U.S. patent No. 4,,73;7,323 for Liposome Extru-sion issued April 12, 1988. Homogenization methods are also useful for down-si2ing li, osomes to sizes of l00nm or less (~artin).
C. ComE~ound Loading In one embodiment, the composition of ~e inventlon is used ~or loc~11 71n~ an imaglng agent, slch as radio-isotopes lncluding '7Ga and ~11In, or parama~netLc co.~-pounds at the tumor site. In this application, where the r~A1 O1 ahe1 can be detected at relatively low concentra-tion, it is generally sufficient to encapsulate the imaging agent by passive loading, i.e., during liposome formation. This may be done, for example, by hydrating lipids wlth an aqueous solutlon of the agent to be encap-sulated. Typically radiolabeled agents are radioisGtopic metals in ~h~ ted form, such as '7Ga-desferal, and are re~ained in the liposomes substantially ~ in entrapped form. After liposome formation and sl2lng, non-encapsu-lated material may be removed by one of a varlety of __ ~, , WO 9l/05j46 PCr/US9O/06211 ~ ~67178 methods, such as by ion exchange or gel filtration chro-matography. The concentration of chelated metal which can be achieved by passive loading is limited by the cnnr~n~ration of the agent in the hydrating medium.
Active loading of radioimaging agents is also pos-sible by entrapping a high affinity, water soluble chela-ting agent ~such as EDTA or desferoxamine) within the aqueous compartment of liposomes, removing any unen-trapped rhf~l at I nrJ agent by dialysis or gel exclusion column chromatography and incubating the liposomes in the presence of t~.e metal radioisotope chelated to a lower affinity, lipid ~oluble chelating agent such as 8-hydr-oxyriuinoline. The metal radioisotope is carried into the liposome by the lipid soluble chelating agent. Once in the liposome, the radiDisotope is chelated by the en-trapped, water soluble rhr~l ~t; ng agent - effectively trapping the radioisotope in the liposome interior (Gabi-zon) .
~assive loading may also b~ employed .'or the ~ ra~h~ c anti-tumor compounds, such as the alkaloids vinblastine and vincristine, which are the-~peutically active at relatively low drug doses, e.g., abou~ 1-15 mg/m2. Here the drug is either dLssolved in the ariueous phase used to hydrate the lipid or included with the lipids in liposome formation process, depending on the solubility of the compound. After liposome formation and slzing, free ~unbound) drug can be removed, as above, for example, by ion exchange or gel exclusion chromatographic methods .
Where the a~ti-tumor compound includes a peptide or protein drug, such as intrr1~1lkin-2 (IL-2) or tissue necrosis factor (TNF), or where the liposomes are formu-lated to contain a peptide immunomodulator, such as muramyl di- or tri-peptide derivatives or a protein WO 91/05546 PCr/US90/06211 ~ 6~ 22 immunomodulator such as macrophage colony _stimulating ~actor (M-CSF), the liposomes are preferably prepared by the above reverse phase method or by rehydrating a freeze dried mlxture of ~ t~e prptein and a sus~ension of small 5 unilamellar vesicles with water ~Kirby). Both methods combine passive loading with relatively high encapsu-lation efficiency, e.g., up to 50% efficiency. Nonencap-sulated materl~al can be readily removed ~rom the liposome suspension, e.g., by dialysis, diafiltratlon or exclusion lO chromatography.
~ he conc~ntr~t~nn o~ hydrophobic drug which can be accommodated in the liposomes will depend on drug/lipid interactions in the membrane, but is generally limited to a drug c~n- ~ntration of less than about 20 ~g drug/mg 15 lipid. More specifically, for a variety o~ anthracycline antibiotics, such as doxorubicin and epirubicin, the highest concentration of encapsulated material which can be achieved by passive loading intD the aqueo~s compart-ment of the liposome is about 10-20 Ils~umoles li}-id ~due 20 to the low intrinsic water solubi' ity of these compounds). When 20-30 mole percent of an a.iionic phos-pholipid such as PG is included in the membra}~e the loading factor can be increased to about ~ g/umole lipid because the anthracyclines are positively charged 2~ and form an "ion pair" complex with the negatively charged PG at the membrane interface. However, such charged complexed anthracycline form111at1nnc have limited utility in the context of the present invention ~which requires that the drug be carried through the bloodstream 30 ~or the first 24-48 hours following IV administration in liposome entrapped form) because the drugs tend to be rapidly released from the liposome membrane when intro-duced into plasma.
.
-In accordance with another aspect o~ the inventionl it has been found essential, for delivery of an therape,~-tically ef~ective dose of a variety of amphipathic anti-tumor drugs to tumors, to load the liposomes to a high drug concentration by active drug loading methods. For exat~ple, for anthracycline antibiotic drugs, such as doxorub~ cin, epirubicin, daunorubicin, carcinomycin, N-acetyladriamYCin, rubidazone, S-;mi~n~ nr ycin, and N-acetyldaunomycin, a final concentration o~ liposome-entrapped drug of greater than about 25 ~g/umole lipid and preferably 50 ~lg/umole lipid is desired. In~ernal drug cnn~ntrations as high as 100-200 ug/umole lipid are contemplated.
one method for active loading of amphipathic drugs into liposomes is described in co-owned U. S .
Patent ~o. ~,192,549. In this method, liposome~ are prepared in the pre~ence o E
a relatively high concentration of ~ ion, such as 0.125 ~ n sulfate~. After sizil:g the liposomes to a desired size, the llposome suspension is treated to create an inside-to-outside ammonium ion g. a~'ient across the liposomal membranes. The gradient may be crea.ed by dialysis against a non-ammonium ron~in;ng .nedi~m, such as an isotonic glucose medium, or by gel filtra~ion, such as on a Sephadex~ G-50 column equilibrated with 0.15~ NaCl or RCl, effectively replacing ammonium ions in th~ exte-rior phase with sodium or potassium ions. Alternat~;ely, the l 1ros suspension may be diluted with a non-am-monium solution, thereby reducing the exterior-phase cnn~ntration of ammonium ions. The ~ rn concen~ra-tion inside the liposomes is preferably at least 10 times, and more preferably at- least 100 to 1000 times that in the external liposome phase.
~Tradema~k ~,,.
- - -WO 91/05546 PCr1US90106~11 6t~
The ammonium ion graaient across the liposomes in turn creates a pH gradient, as ammonia is released across the liposome membrane,_and protons are trapped in the internal aqueous phase, of the liposome. To load lipo-somes wlth the selected drug a suspenslon of the lipo-somes, e.g., about 20-200 mg/ml lipid, is mixed with an aqueous solution of the drug, and the mixture is allowed to equilibrate over_ an period of time, e.g., several hours, at temperatures ranging from room temperature to 6~C - depending on the phase transition temperature of the lipids used to form the liposome. In one typical method, a suspension of liposomes having a lipid con-centration of 50 umoles/ml is mixed with an equal volume of anthracycline drug at a concentration of about 5-~
I5 mg/ml. At the end of the incubation period, the suspen-sion is treated to remove _ree (unbound) drug. One preferred method of drug removal for anthracycline drugs is by passage over an ion exchange resin, such ~s Dowex 50 WX-4, which is capable of binding ti~e drug.
Although, as noted above, the plant a ' kaloids such as vincristine do not require high loading factors in liposomes due to their intrinsically high anti-tumor activity, and thus can be loaded by passiv2 ~ntrapment techniques, it also possible to load these drug by active methods. Since vincristine is amphipathic and a weak base, it and similar molecules can be loaded into lipo-somes using a pH gradient formed by entrapping ammonium sulfate as described above for the anthracycline antibio-tics .
The remote loading method just described is il ` us-trated in Example l0, which descrlbes the preparation of 0.1 micron ~LVs loaded with doxorubicin, to a final drug concentration of about 80-lO0 ~Lg/umoles Iipid. The lipo-. _ = ~
WO 91/05546 PCr/US90/06211 .
20671'~{8 somes show a very low rate of drug leakage when stored at III. Liposome Localization in Solid Tumors A. ~rton~1~d Bloodstream Halflife One of the requirements for liposome localization in a target tumor, in accordance wlth the inventlon, is an exten~ed liposome lifetime~ ln the bloodstream following IV lipo-~ome administration. one measure of liposome lifetime in the bloodstream in the blood/RES ratio deter-mined at a selected time after liposome administration, as discussed above. Blood/RES ratios for- a variety of liposome compositions are given in Table 3 of Example 5.
In the absence of PEG-derivatized lipids, blood/RES
ratios were 0 . 03 or less . In the presence of PEG-deriva-tized iipids, the blood/RES ratio ranged from 0.2, for low-molecular weight PEG, to between l . 7-4 for several of the formulations, one of which lacks cholesterol, and three of which lack an added charged phospholipid (e.g., PG).
The data presented in Table 5 in Exa~ple 6 show blood/RES ratios ~excluding two points with low percent recovery) between about 1.26 and 3.27, cor si;t~nt with the data given in Table 3. As noted in Section II above, the blood lifetime values are subst~nt; ~1 1y independent of degree of saturation of the liposome lipids, presence of cholesterol and presence of charged lipids.
The blood/RES values reported above can be compared with blood/RES values reported in co-owned U . S . Patent No. 4, 920, 016, whiCh used blood/RES mea~uL~ -nt methods identical to those used in generating the data presented in Tables 3 and 5. The best 24-hour blood/RES ratios which were reported in the above-noted patent was 0 . 9, for a formulation composed of GMI, saturated PC, and 26 2~67 1 78 cholestero~. The next best formulations gave 24-hour blood/RES values of about 0 . 5 . Thus, typical 24-ho~ur blood/RES ratios ob-ained in a number of the current formulations were more than twice as high as the best 5 formulations which have been reported to date. Furthe-, ability to achieve high blood/RES with GMI or HPI lipids was dependent on the presence of prednr~i nAntly saturated lipids and cholesterol in the liposomes.
Plasma phArm~cokinetics of a liposomal marker in the lO bloodstream can provide another measure of the ~nhAnCPd liposome lifetime which is achieved by the liposome formulationS of the present invention. Figure~ 7 and 8 discussed above show the slow loss of liposomal marker from the bloodstream over a 24 hour period in typical 15 PEG-liposome form~1At;ons, substAnt jA1~y ~n~iPpen~Pnt of whether the marker is a lipid or an encArsul ~ted water-soluble compound lFigure 8). I;l both plots, the amount of liposomal marker present 24 ~ours after liposome injection is greater than 10% of the ~riginally injected 2 0 material .
Figure 9 shows the kinetics of liposom~ loss from the blood stream for a typical PEG-liposom0 form~:lation and the same liposomes in the absence of a ~r ' -deri~-a-tized lipid. ~fter 24 hours, the percent marker remain-25 ing in the PEG-liposomes was greater than about 2096, whereas the conventional liposomes showed less than 5%
retention in the blood after 3 hours, and virtuallY no detectable marker at 24 hours.
The results seen in Figures 7-9 are consistent with 30 24 hour blood liposome values measured for a variety of liposome formulations, and reported in Tables 3 and 5-7 in Example 5-8 below. As seen in Table 3 in Exam?le 5, the percent dose rPr~-;n;n~ at 24 hours was less than 1%
for conventional liposomes, versus at least 5% for -he A
PCI/US90/062tl WO 91/05~46 , , .
2067~7~ -PEG-liposomes. In the best form~ f 1 ons, values between about 20-40~6 were obtained. Similarly in Table 5 from - Example 6, liposome levels in the blood after 24 - hours (again neglecting two entries with low recovery values) 5 were between 12 and about 25 percent of total dose given.
Similar results are reported in Tables 6 and 7 of Example 7.
The ability of the liposomes to retain an amphi-pathic anti-tumor drug in the bloodstream over the 24-48 perlod required to provide an opportunity for the lipo-some to reach and enter a systemic tumor has also been investigated. In the study reported in Example ll, the plasma ~h~rm~sk~n~otics of doxorubicin loaded in PEG-liposomes, doxorubicin ~riven in free form, and doxorubi-cin loaded into liposomes contalning hydrogenated phos-phatidylinositol ~iPI) was in~ested in beagle dogs. The ~IPI liposomes were formulated wi'h a pre~ ~ n;lnt1y satu-rated PC lipid and cholesterol, and represents one of the optimal fQr~lAtion5 descr$bed in the above co-owr.ed U.S.
patent. The kinetics of doxorubicin in the blood up to 72 hours after drug administration is shown ir Figure ll.
Both liposomal fo lat~ons showed single-rrLode exponen-tial loss of drug, in contrast to free drug ~ h ~ 'i shows a bi-exp~n~ont ~ ~1 pattern . However, the amount of drug retained in the blood stream at 72 hours was about 8-10 times greater ln the PEG-liposomes.
For both blood~RES ratios, and liposome retention time in the bloodstream, the data obtained from a model animal system can be reasonably extrapolated to humans and veterinary animals of interest. This is because uptake of liposomes by liver and spleen has been f ound to occur at similar rates ln several mammalian species, including mouse, rat, monkey, and human (Gregoriadis, 1974; Jonah: Kimelberg, 1976, Juliano, Richardson;
. _ , ~
~ope~-Beresteir.). This result likely reflects the fact that the biochemical factors which appear to be m~st important in liposome uptake by the RES -- including opsinization by serum lipoproteins, size-dependent uptake 5 effects, and cell shielding by surface moieties -- are common features of all mammalian species which have been t'~Aml nf~d.
B. Extravasation into Tumors Another required feature for high-activity liposome targeting to a solid tumor, in accordance with the inven-tion, is liposome extravasation into the tumor through the endothelial cell barrier and underlying basement membrane separating a capillary from the tumor cells 15 supplied by the capillary. This feature is optimized in liposomes having sizes between 0 . 07 and 0 .12 microns .
That liposome delivery to the tumor is required for selective drug targeting can be seen from the study reported in Example 12. Here mice were inoculated sub-20 cutaneously with the J-6456 lymphoma whic~ formed a solid tumor mass of about 1 cm3 after one-two we~ks~ The ani-mals were then injected either with free dcxorubicin or doxorubicin loaded into PE&-liposomes at a :lo-s~ of l~m~
drug per kg body weight. The tissue distribution (heart, 25 muscle, and tumor) of the drug was then assayed at 4, 241 and 48 hours after drug administration. Figure llA shows the results obtained for free drug. No selective drug A~ m~ on into the tumor occurred, and in fact, the highest initial drug levels were in the heart, where 30 greateSt toxicity would be produced.
By contrast, drug delivery in PEG-liposomes showed increasing drug acc~m--l Ation into the tumor between 4-24 hours, and high selective tumor levels between 24 and 48 hours. Drug uptake by both heart and muscle tissue was, A
by contrast, lower than with free drug. As seen from the data plotted in Figure llB, the tumor cont~ined 8 ti~eS
more drug compared with healthy muscle and 6 times the amount in heart at 24 hours post injection.
To confirm that the PEG-liposomes deliver more anti-tumor drug to a intraperitoneal tumor, groups of mice were injected IP with 10~ J-64S6 lymphoma cells. After five Idays the IP tumor had been established, and the animals were treated IV with lOmg/kg doxorubicin, either in free drug form or entrapped in PEG-cont~;n;n~ lipo-somes. Tlssu~ distribution of the drug is tabulated in Table 9, Example 12. As shown, the tumor/heart ratio was about 272 greater for liposome delivery than for free drug at 24 hours, and about 47 times greater at 48 hours.
To demonstrate that the results shown in Table 9 are due to the entry of intact liposomes into the extravas-cular region of a tumor, the tu~or tissue was separated into cellular and s~rernPt~nt 'intercellular fluid) fractions, and the presence of liposome-associ.~ted and free drug in both fractions was assayed. Figure 12 shows the total amount of drug (filled ~ mr nr~ anc the amount of drug present in tumor cells (solid circle~) and in the supernatant in liposome-associated form (~olid triangles) over a 48-hour post injection period. To assay liposome-associated drug, the super-25 natant was passed through an ion-exchange resin to remove free drug, and the drug L. ~ I n~ ng in the supernatant was assayed (solid triangles). As seen, most of the drug in the tumor is liposome-associated.
Further demonstration of liposome extravasation into 30 tumor cells was obtained by direct microscopic observa-tion of liposome distribution in normal liver tissue and in solid tumors, as ~3~t~ led in Example 14. Figure 13A
shows the distribution of liposomes (small, darkly stained bodies) in normal liver tissue 24 hours after IV
injection of P~G-liposomes. The liposomes are confined exclusively to the KuDfer cells and are not prese~t either in hepatocytes or in the intercellular fluid ~f the normal liver tissue.
Figure 13B shows a region of C-26 colon carcinoma implanted in the liver of mice, 24 hours after injection of PEG-liposomes. Concentrations of liposomes are clear-ly evident in the region of the capillary in the figure, on the tumor tissue side of the endothelial barrier and basement membrane. Liposomes are also abundant in the intercellular fluid of the tumor cells, further eviden-cing passage from the capillary lumen into the tumor.
The Figure 13C photomicrograph shows another region of the tumor, where an abundance of liposomes in the inter-cellular fluid is also evident. A similar finding was made with liposome extravasation into a region of C-26 colon carcinoma cells injected sl~hcl~t~n~ously, as seen in Figure 13D.
IV. Tumor Localization ~ethod As detailed above, the liposomes of th~ invention are ef fective to localize specifically in a ~olid tumor region by virtue of the extended lifetime of ' he lipo-somes in the bloodstream and a liposome size which allows both extravasation into tumors, a relatively high drug carrying capacity and minimal leakage of the entrapped drug during the time required for the liposomes to dis-tribute to and enter the tumor (the first 24-48 hours following injection). The liposomes thus provide an effective method for loc~l;7;ng a compound selectively to a solid tumor, by entrapping the compound in such lipo-somes and injecting the liposomes ~V into a subject. In this context a solid tumor is defined as one that grows n ~n~omlc-l sl~e outslde the bloodstre~m (ln ~on-WO 91/0~546 PCr/US90/06211 2o67l78 trast, for example, to blood-born tumors such as leuke-mias) and requireS the formation of small blood vessels - - and capillaries to supply nutrients, etc. to the growing tumor mass. In this case, for an IV injected liposome - 5 (and its entrapped anti-tumor drug) to reach the tumor site it must leave the bloodstream and enter the tumor.
In one: -~;r L, the method is used for tumor treatment by lor~1l7in~r an anti-tumor drug selectively in the tumor. The anti-tumor drug which may be used is any compound, including the ones listed below, which can be stably entrapped in liposomes at a suitable loading factor and administered at a therapeutically effective dose (indicated below in parentheses after each compound) . These include ; h I r~h ~ c anti-tumor com-pounds such as the p~ ant alkaloids vincristine ~1. 4 mg/m2), vinblastine ~4-18 mg/m2) and etoposide (35-100 mg/m2), and the anthracycline antibiotics including doxo-rubicin (60-75 mg/m2), epirubicin (60-120 mg/m2) and daunorubicin (25-~5 mg/m2). The water-~soluble anti-meta-bolites such as methotrexate 3 mg/m2), c~tosine arabino-side (100 mg/m2), and fluorouracil (10-lS m3/kg), the antibiotics such as bleomycin (10-20 units/m2), mitomycin (20 mg/m2), plicamycin (25-30 ug/m2) and dactinc li~cin ;15 ug/m2), and the alkylating agents includlng cyclophospha-mide (3-25 mg/kg), thiotepa (0 . 3-0 . 4 mg/Kg) and BCNU
(150-200 mg/m2) are also useful in this context. Æs noted above, the plant alkaloids exemplified by vincris-tine and the anthracycline antibiotics including doxoru-bicin, daunorubicin and epirubicin are preferably active-ly loaded into liposomes, to achieve drug/lipid ratios which are several times greater than can be achieved with passive loading. Also as noted above, the liposomes may contain encapsulated tumor-therapeutic peptides and protein drugs, such as IL-2, andtor TNF, and/or immano-modulators, such as M-CSF, which are present alone or; in ccmbination with anti-tumor drugs, such as an anthraa~y-cline antibiotic drug.
The ability to ef~ectively treat solid tumors, in 5 accordance with the present invention, has been shown in a variety of in vivo systems. The method reported in Example 15 compares the rate of tumor growth in animals with ir~planted subcutaneously with a C-26 colon carci-noma. Treatment was with epirubicin, either in free 10 form, or entrapped in PEG-liposomes, in accordance with the invention, with the results shown in Figures 14A-C.
As seen, and discussed more fully in Example 15, treat-ment with epirubicin loaded PEG-liposomes produced a marked supression of tumor growth and lead to long term 15 survivors among groups of animals inoculated with a normally lethal dose of tumor cells. Moreover, delayed treatment of animals wlth the epiribicin loaded PEG lipo-somes resulted in regression of est~hl ~ ched subcutaneous tumors, a result not seen with free drug treatmen..
Similar results were obtained for treatment of a lymphoma implanted interperitoneally in mice, ~a~s detailed in Example 16. Here the animals were treate.~ with doxo-rubicin in free form or entrapped in P~3G- :- 70som~s .
Percent survivors over a 100-day period following tumor impl~n~isn and drug treatment is shown in Figure 16.
The results are similar to those obtained above, showing marked increase in the median survival time and percent survivors with PEG-liposomes over free drug treatment.
Since reduced toxicity has been observed in model animal systems and in a ~~lnic~l setting in tumor t3:eat-ment by doxorubicin entrapped in conventional liposomes ~as reported, for example, in U.S. Patent No. 4,898,735), it is of interest to determine the degree of toxicity protection provided in the tumor treatment method of the present invention. In the study reported ln Example 17, animals were injected Iv wlth increasing doses of doxo~ru-bicin or epirubicin in free form or entrapped in conven-tional or ~EG-liposomes, The maximum tolerated dose 5 ~TD) for the various drug formulations is given in Tzble lO in the Example. For both drugs, entrapment in PEG-liposomes appro~ t~l y doubled the ~TD of the drug .
Similar protection was achieved with conventional lipo-somes .
~th''U~Th reduced toxicity may contribute to the increased efficacy o~ tumor treatment reported above, selective lorll;7~ti~n of the drug by liposomal extrava-sation is also important for improved drug efficacy.
This is demonstrated in the drug treatment method de-15 scribed in Example 18. E~ere conventional liposomes cnnt~;n~ng doxorubicin (which show little or no tumor uptake by extravasation when administered IV) were com-pared with free drug at the sam~ dose (lO m~tkg) to reduce reduce the rate of growth of a subcuat i.neously 20 implanted tumor. Figure 16 plots tumor s~.ze with time in days following tumor implantation for a sal~ne control ~solid line), free drug (filled circles) and -o~ventional liposomes ~filled triangles) . As seen conver ti~nal l_po-somes do not supress tumor growth to any greater ~xtent 25 than free drug at the same dose. This finding stands in stark contrzst to the results shown in Figures 14A-C and 15 where improved survival and tumor growth supression is seen compared to free drug when tumor-bearing animals are treated wlth anthracycllnes anti-tumor drugs entrapped in 30 ~EG l; ros s .
Thus, the tumor-treatment method allows both higher levels of drug to be administered, due to reduced drug toxicity in liposomes, and greater drug efficacy, due to selective liposome localization in the intercellular A
WO 91/05546 Pcr/us9o .r fluid of the tumor.
It willj be appreciated that the ability to locali2e a compcund selectively in a tumor, by liposome extravasa-tion, can also be exploited for improved targeting of an 5 imaging agent to a tumor, for tumor diagnosis. Here the imaging agent, typically a radioisotope in chelated form, or a paramagnetic molecule is entrapped in liposomes, which are then administered IV to the sub ject being PxAm; nec~ . After a selected period, typically 24-48 10 hours, the subject is then monitored, for example by gamma scintillation radiography in the case of radioiso-tope or by N~qR in the case of the paramagnetic agent, to detect regions of local uptake of the imaging agent.
The following examples illustrate methods of 15 preparing liposomes with enhAn~P~ circulation times, and for accPssing circulation times in vivo and in vitro.
The examples are ~ntPn~led to illustrate srPr~f~c liposome compositions and methods of the inv~ntion, but are in no way intended to limit the scope thereof.
Materials Cholesterol (Chol) was obtained from Sigma (St.
Louis, NO) . Sphingomyelin (SN), egg phosphati~y] chol ne (lecithin or PC), partially hydrogenated PC havins the 2~ composition IV40, IV30, IV20, IV10, and IV1, phosphati-dylglycerol (PG), phnsph~tldylethanolamine (PE), dipalmi-toyl-phosphatidyl glycerol (DPPG), dipalmitoyl PC (DPPCl, dioleyl PC (DOPC) and distearoyl PC ~DSPC) were obta' ned from Avanti Polar Lipids (~irm~ngh~m, AL) or Austin 30 Chemical Company ~Chicago, IL).
["sI]-tyraminyl-inulin was made according to pub-lished procedures. 67Gallium-8-hyd,oxy~luinolLne was sup-plied by NEN Neoscan ~Boston, NA). Doxorubicin E~Cl and Epirubicin HCL were obtained from Adria Laboratorles (Colu;n~us. OH) or Farmitalia Carlo Erba (Mil2n, Italy) .
Example 1 Pre~aration of PEG-PE Linked by Cyanuric C h l o -5 ride A. Preparation of activated P~;G
2-0-Methoxypolyethylene qlycol 1900-4, 6-dichlo-ro-l,3,5 triazine previously called activated PEG was prepared as described in J. Biol. Chem., 252:3~82 ~1977) l0 with the following mo~fir~ons.
Cyanuric chloride (5.5 g; 0.03 mol) was dissolved in 400 ml of anhydrous benzene cont~;n;n~ 10 g of anhydrous sodium c~rh~"Ate, and PEG-1900 ~19 g; 0.01 mol) was added and the mixture was stirred overnight at room tempera-15 ture. The solution was ~iltered, and 600 ml of petroleumether (ho~ t ~ ng range, 35-60O) was added slowly with stir-ring. The ~inely divided precipitate was collected on a filter and redissolved in 400 ml o~ benzene. T~.e preci-pitation and ~iltration process was repeated several 20 times until the petroleum ether was free of residual cyanuric chloride as l~t~ n--d by high pres~-~re liquid chromatography on a column ~250 x 3.2 mm) of S-m "~i~hro-orb~" ~E. ~erck), developed with hexane, and det~ed with an ultraviolet detector. Titration of activated 25 PEG-1900 with silver nitrate after overni~ht hydrolysis in aqueous buffer at pH 10 . 0, room temperature, gave a value of 1. 7 mol of chloride liberated/mol of PEG.
T~C analysis of the product was effected with T~C
reversed-phase plates obtained from Baker using methanol-30 water, 4:1; v/v, as developer and exposure tO iodinevapor for vis~ Ation. Under these conditions; the startinq methoxy polyglycol 1900 appeared at R~=0.54 to 0 . 60 . The activated PEG appeared at Rf=0 . 41. Unreacted cyanuric chloride appeared at Rf=0 . 88 and was removed.
~Trademark A
WO 91/05~46 PCr/US90/06211 .~
The actlvated PEG was analyzed for nltrogen and an appropriate correctlon was applied ln selecting the quantity of reactant to use in further synthetic steps.
Thus, when t~he product contained only 20% of the theore-5 tical amoui~t of nitrogen, the quantity of material usedin the next synthetic step was increased by 10 0 /2 0, or 5-fold. When the product c-~nt~ 1 n~ 50% of the theore-tical amount of nltrogen, only 100/S0 or a 2-fold in-crease was needed.
B. Preparation of N- (4-Chloro-polyglycol 1900~-1,3,5-triazinyl egg phosphatldylethanolamine.
In a s~ ed test tube, 0.74 ml of a 100 mg/ml (0.100 mmole) stock solution of egg phosphatidylethanol-15 amine ln chloroform was evaporated to dryness under astream of nitrogen ana was added to the residue of the activated PEG described in secti on A, in the amount to provide 205 mg (0.100 mmole). To 1:his mixture, 5 ml an-hydrous dimethyl forTn~m1 rlP was added. 27 microliters 20 (0.200 mmole) triethylamine was added to ~he mixture, and the air was displaced with nitrogen gas. The ~.ixture was heated overnight in a sand bath maintained at 110C.
The mixture was then evaporated to cryn~ss ~ er vacuum and a pasty mass of crystalline solid was ob-25 tained. This solid was dissolved in 5 ml of a mixture of4 volumes of acetone and 1 volume of acetic acid. The resulting mixture was placed at the top of a 21 mm X 2gO
mm chromatographic absorption column packed with silica gel (~erck E~ieselgel 60, 70-230 mesh) which had first 30 been moistened with a solvent composed of acetone ac~tic acid, 80/20; v~v.
The column chromatography was developed with the same solvent mixture, and separate 20 to 50 ml aliquots of effluent were collected. Each portion of effluént was . _ .
.
WO 9l/05546 PCr/US90/06211 ~ 20871,7-~
2ssayed by lLC on silica gel coated plates, using 2-buta-none/acetic acid/water; 40/25/5; v/v/v as developer and iodine vapor exposure for visualization. Fractions containing only material of R~=about 0.79 were combined - 5 and evaporated to dryness under vacuum. Drying to con-stant weight under high vacuum afforded 86 mg (31. 2 micromoles) of nearly colorless solid N- ~4-chloro-poly-glycol 1900)-1,3,5-triazinyl egg phosphatidylethanolamine Cont 1 l n; n~ phosphorous .
The solid compound was taken up in 24 ml of etha-nol/chloroform; 50/50 chloroform and centrifuged to remove insoluble- material. Evaporation of the clarified solution to dryness under vacuum afforded 21 mg (7 . 62 micromoles) of colorless solid.
Example 2 Preparation of C~rh~m~te and Amide Linked Hydrophilic Polymers with PE
A. Preparation of the imidazole r~rh~m te cf poly-20 ethylene glycol methyl ether 1900.
9.5 grams (5 mmoles) of polyethylene gly;ol methylether l900 obtained from Aldrich Chemical Cc. was dis-solved in 45 ml benzene which has been drie ~ ov~r mo' e-cular sleves. 0.89 grams ~5.5 mmoles) of pure carbonyl 25 ~; im~rl~7ole was added. The purity was checked by an infra-red spectrum. The air in the reaction vessel was displaced with nitrogen. Vessel was enclosed and heated in a sand bath at 75C for 16 hours.
The reaction mixture was cooled and the clear solu-30 tion formed at room temperature. The solution was ~ilu-ted to 50 . 0 ml with dry benzene and stored in the refri-gerator as a 100 micromole/ml stock solution of the imidazole carbamate of PEG ether 1900.
WO 91/05546 PClr/US90/06211 ~S ~ ~ ~ _ ~ 6¢1 38 B. Preparation of the phosphatidylethanolamine car-bamate of polyethylene glycol methyl ether l900.
lO . 0 ml ~lmmol) of the lO0 mmol/ml stock solution of the imidazole carbamate of polyethylene glycol methyl ether l900 was pipetted lnto a lO ml pear-shaped flask.
The solvent was removed under vacuum. 3.7 ml of a lO0 mg/ml solutlon of egg phosphatidyl ethanolamine in chlo-roform (0.5 mmol) was added. The solvent was evaporated under vacuum. 2 ml of l, l, 2, 2-tetrachloroethylene and 139 microllters (l.0 mmol) of triethylamine VI was added.
The vessel was closed and heated in a sand bath main-tained at 95C for 6 hours. At this time, thin-layer chromatography was performed with fractions of the above mixture to determine an extent of con~ugation on Sl02 coated TLC plates, using butanone/acetic acid/water;
40/5/5; v/v/v; was performed as developer. I2 vapor V; Sl~A 1 i 7At ~ on revealed that most of the free phosphatidyl ethanolamine of Rf=0 . 68, had reacted, and was replaced by a phosphorous-c~ tA~n1n~ lipid at R~sO. ,8 to 0.80.
The solvent from the l~ ~ning reaction mixture was evaporated under vacuum. The residue was take.l up in lO
ml methylene chloride and placed at the top oE a 21 mm x 270 mm chromatographic absorption column pac;-~d w th ~erck Rieselgel 60 (70-230 mesh silica gel), which has been first rinsed with methylene chloride. The mixture was passed through the column, in sequence, using the following solvents.
-.
Table 1 Volume % of Volume % Methanol ml Methylene Chloride With 2% Acetic Acid 5~00 100%
200 95% 5%
200 90% 10%
200 85% 15%
200 60% 40%
50 ml portions of effluent were collected and each portion was assayed by TLC on SiO2 - coated plates, using 12 vapor absorption for v; c~ ; 7at inn after developmen~
with chloroform/methanol/water/c~nrPntrated ammonium hydroxide; 130/70/8/0.5%; v/v/v/v. Most of the phos-phates were found in fractions 11, 12, 13 and 14.
These fractions were ' ine~l, evaporated to dryness under vacuum and dried in high vacuum to constant weight.
They yielded 669 mg of colorless wax of phosphatidyl 20 etha-nolamine r~rhA~ e of polyethylene glycol methyl ether. This represented 263 ~icromoles and a vield of 52.6% based on the rhosE~h~t;~yl ethanolamine.
An N~R spectrum of the product dissol~-ed in deutero--chloroform showed peaks corresron~l; ng to the s~ctrum for 25 egg PE, together with a strong singlet due to the methy-lene groups of the ethylene oxide chain at Delta = ~ . 4 ppm. The ratio of methylene protons from the etr~ lene oxide to the t~rm;n~l methyl protons of the PE acyl - groups was large enough to confirm a molecular weight of 30 about 2000 for the polyethylene oxide portion of the molecule of the desired product polyethylene ~ col conjugated phosphatidyethanolamine c;~rh~ te, M.W. 2, 654 .
C. Preparation of polylactic acid amide of phosphotl-dyletanolamine .
A
,a_ WO 91/05546 PCr/US90/06211 __ - 40 200 mg (0.1 mmoles) poly (lactic acid), m. wt. s 2, 000 (ICN, Cleveland, Ohio) was dissolved in 2.0 ml dimethyl sulfoxide by heating while stirring to dissolve the material completely. Then the solutlon was cooled imme-diately to 65 C ~ and poured onto a mixture of 75 mg (0.1 mmoles) of distearylphosphatidyl-ethanolamine (cal.
Biochem, La Jolla) and 41 mg (0.2 mmoles) dicyclohexyl-carbodiimide. Then 28 ml (0.2 mmoles) of triethylamine was added, the air swept out of-the tube with nitrogen gas, the tube capped, and heated at 65C for 48 hours.
After this time, the tube was cooled to room tempera-ture, and 6 ml of chloroform added. The chloroform solution was washed with three s~lr~r~sc1~e 6 ml volumes of water, centrifuged after each wash, and the phases sepa-rated with a Pasteur pipette. The I` ; n; ng chloroform phase was filtered with suction to remove suspended distearolyph~srhAt ~ ~ylethanolamine . The filtrate was dried under vacuum to obtain 212 mq of semi-crystalline solid .
This solid was dissolved in 15 ml o~ a mixture of 4 volumes ethanol with 1 volume water and passed through a 50 mm deep and 21 mm diameter bed of H' Dowex c o cation exchange resin, and washed with 100 ml of the salr.e ~o,l-vent .
The filtrate was evaporated to dryness to obtain 131 mg colorless wax.
291 mg of such wax was dissolved in 2.5 ml chloroform and transferred to the top of a 21 mm x 280 mm colu.,ln of sLlica gel wetted with chloroform. The chromatogram was developed by passing through the column, in sequence~ 100 ml each of:
100% chloroform, 0% (1% NH,OH in methanol);
90% chloroform, 10% (1% NE~OH in methanol);
85% chloroform, 15% (1% NH~OH in methanol), , WO 91/05546 PCr/US90/06Zl I
2~67178 80% chloroform, 2Q9~; (196 NH,OH in methanol);
70% chloroform, 30% (1% NH~OH in methanol);
Individual 25 ml portions of effluent were saved and assayed by TLC on SFOz-coated plates, using CHCl3, CH,OH, H70, con. NH~OH, 130, 70, 8, 0.5 v/v as developer and I2 vapor absorption for visualization.
The 275-325 ml portions of column effluent contained a single material, PO, +, of R~ = 0 . 89 .
When c~ ' in~d and evaporated to dryness, these afforded 319 mg colorless wax.
Phosphate analysis agrees with a molecular weight of possibly 115, 000 .
Apparently, the polymerization of the poly (lactic acld) occurred at a rat~ comparable to that at which lt reac~ed with phosphatidylethanolamine.
This side-reaction could probably be minimized by working with more dilute solutions of the reactants, D. Preparation of poly (glycolic acid) amide of DSPE
~ mixture of 266 mg. ~3.50 mmoles) glycolic acid, 745 mg (3.60 mmoles) dicyclohexyl carbodiimide, 75 mg. (0.10 mmoles) distearoyl phosphatidyl eth~n~ ml n~, ~? mi~-o-liters (0.23 mmoles triethyl amine, and 5.0 ml dry ~im-ethyl sulfoxide was heated at 75 C, under a nitrogen atmosphere, cooled to room temperature, then diluted with an equal volume of chloroform, and then washed with three successive equal volumes of water to remove dim~thyl sulfoxide. Centrifuge and separate phases wit~ a Pasteur pipette each time.
Filter the chloroform phase with suction to remove a small amount of suspended material and vacuum evaporate the filtrate to dryness to obtain 572 mg. pale amber wax.
WO 9I/05546 PCr/US90/06211 6'~ ~ 42 Re-dissolve this material in 2 . 5 ml chloroform and transfer to the top of a 21 mm X 270 mm column of silica gel (Merck Hieselgel 60I which has been wetted with chloroform .
Develop the ~ hromatogram by passing through the column, in se~uence, 100 ml each of: ~
100% chloroform, 0 % tl% NH,OH in methanol);
90% chloroform, 1095 (1% NHIOH in methanol);
85% chloroform, 15% (1% NH~OH in methanol);
80% chloroform, 20% (1% NH~OH in methanol);
70% chloroform, 30% (1% NH~OH in methanol) .
Collect individual 25 ml portions of effluent and assay each by TLC on Si) 2-coated plates, using CH Cl3, CH3 OH, H20, con-NE~OH; 130, 70, 8, 0.5 v/v as developer.
Almost all the PO4 + material will be in the 275-300 ml portion of effluent. Evaporation of this to dryness under vacuum, followe~ by high-vacuum drying, affords 281 mg of colorless wax.
ph~srh~t,~ analysis suggests a molecular w6ight of 924, 000 .
Manipulation of solvent volume during re~ction and molar ratios of glycolic acid and dicyclohexyl carbodi-imide would probably result in other sized molecul es .
Example 3 Preparation of Ethylene-Linked PEG-PE
A. Preparation of I-trimethylsilyloxy-polyethylene glycol is illustrated in the reaction scheme sho~ in Figure 3.
15.0 gm tlO mmoles) of polyethylene glycol) M.Wt. 1500, (Aldrich Chemical) was dissolved in 80 ml benzene . I . 40 ml (11 mmoles) of chlorotrimethyl silane (Aldrich Chemi-cal Co. ) and 1.53 ml (lmmoles) of triethylamine was added. The mixture was stirred at room temperature under ' _ an inert atmosphere for 5 hours.
The mixture was filtered with suction to separate crystals of triethylammonium chloride and the crystals were washed with 5 ml benzene. Filtrate and benzene wash 5 liquids were . ~; ne~l . This solution was evaporated to dryness under vacuum to provide 15 . 83 grams of colorless oil which solidified on standing.
TLC of the product on Si-C1, reversed-phase plates using a mlxture of 4 volumes of ethanol with 1 volume of 10 water as developer, and iodine vapor visualization, revealed that all the polyglycol 1500 (Rt=0 . 93) has been consumed, and was replaced by a material of R~=0 . 82 . An infra-red spectrum revealed absorption peaks characteris-tic only of polyglycols.
Yield of I-trimethylsilyoxypolyethylene glycol, M.W.
1500 was nearly quantitative.
B. Preparation of trifluoromethane sulfonyl ester of ltrimethylsilyloxy-polyethylene glycol.
15.74 grams (10 mmol) of the crystalline I-trLmethyl-20 silyloxy polyethylene glycol obtained abov~ w~s dissolvedin 40 ml anhydrous benzene ar,d cooled in ~ bath of crushed ice. 1.53 ml (11 mmol) triethylamine and 1.85 ml (11 mmol) of trif~ rome~h~n~c~l fonic anhydride qbtained from Aldrich Chemical Co. were added and the mixture was 25 stirred over night under an inert atmosphere until the reaction mixture changed to a brown color.
The solvent was then evaporated under reduced pressure and the residual syrupy paste was diluted to lOû O ml with methylene chloride. Because of the great reactivity 30 of trifluo~o~ h~nP sulfonic esters, no further purif, ca-tion of the trifluoromethane sulfonyl ester of I-tri-methylsilyloxy polyethylene glycol was done.
C. Preparation of N-1-trimethylsilyloxy polyethylene glycol 1500 PE.
_ _ WO 91~05~46 PCr/US90/06211 .
lO ml of the methylene chloride=stock sQlution of the trifluoromethane sulfonyl ester of ;-trimethylsilyloxy polyet};Lylene glycol was evaporated i:Q dryness under - vacuum to obtain about 1.2 grams of residue ~approxi-- 5 mately 0.7 mmoies). To this residue, 3.72 ml of a ch~o-r~:form solution containlng 372 mg (0.5 mmoles) egg PE was added. To the resultLng solution, 139 microliters ~1. 0 mmole) of triethylamine was added and the solvent was evaporated under vacuum. To the obtained residue, 5 ml dry dimethyl f~ m~fl~ and 70 microliters (0.50 mmoles) - triethylamine (VI) was added. Air from the reaction vessel was displaced with nitrogen. The vessel was closed and heated in a sand bath a 110C for 22 hours.
The solvent was evaporated under vacuum to obtain 1.58 grams of brownish colored oil.
A 21 X 260 mm chromatographic absorption column filled with Kieselgel 60 silica 70-230 mesh, was p-epared and rinsed with a solvent composed of 40 volumes of butanone, ~5 volumes acetic acid and 5 volumes of water. The crude product was dissolved in 3 ml of the san~ s?lvent and transferred to the top ~of the chromatograph~ column. The chromatogram was developed with the same solvent and sequential 30 ml portions of effluent were assayed eac~
- ~-by TLC.
The TLC assay system used silica gel coated glass plates, with solvent combination butanone/acetic acid/wa-ter; 40/25/5; v/v/v. Iodine vapor absorption served for ~ v; su~ l i 7ation . In this solvent system, the N-l -tri-methylsilyloxy polyethylene glycol 1500 PE appeared at R,=0.78. Unchanged PE appeared at R~=0.68.
- The desired N-l-trimethylsilyloxy polyethylene glycol 1500 PE was a chief constituent of the~ 170-300 ml por-tions of column effluent. WhFn evapo~ated to dryness .
WO 91/05546 PCr/US90/06211 2~6717~
under vacuum these portions afforded 1~1 mg of pale yellow oil of compound. ~
D. Preparation of N-polyethylene glycyl 1500: phospha-tidyl-ethanolamine acetic acid deprotection.
Once-chromatographed, ~E compound was dissolved in 2 ml of tetrahydrofuran. To this, 6 ml acetic acid and 2 ml water was added. The resulting solution was let to stand for 3 days at 23C. The solvent from the reaction mix-ture was evaporated under vacuum and dried to constant weight to obtaln 75 mg of pale yellow wax. TLC on Si-C18 reversed-phase plates, developed with a mixture of 4 volumes ethanol, 1 volume water, indicated that some free PE and some polyglycol-like material formed during the hydrolysis.
The residue was dissolved in 0 . 5 ml tetrahydrofuran and diluted with 3 ml of a solution of ethanol water: 80:20;
v:v. The mixture was applied to the top of a 10 ~m X 250 mm chromatographic absorption column packed with octade-cyl bonded phase siLica gel and column was deve] oped with ethanol water 80:20% by volume, collecting s~quential 20 ml portions of effluent. The effluent was assayed by reversed phase TLC. Fractions cnntA1n~ng only pro~ct of Rf=0 . 08 to 0 .15 were combined. This was typically the 20-100 ml portion of effluent. When evaporated to dry-- ness, under vacuum, these portions afforded 33 mg of colorless wax PEG-PE corresponding to a yield of only 3%, - based on the starting phosphatidyl ethAnol~mi nP
NMR analysis indicated that the product incorporated both PE residues and polyethylene glycol residues, but that in spite of the favorable-appearing el - Al analy-sis, the chain length of the polyglycol chain has been reduced to about three to four et~llene oxide residues.
WO 91/05546 PCrJUS90/06211 .
c~'~'~ `
The product prepared was used for a preparation of PEG-PE
liposomes. ~; ~
., E. ~ Preparation of N-Polyethylene glycol 1500 P.E. by 5 fluorlde deprotection.
500 mg of crude N-1-trimethylsilyloxy polyethylene gly~ol PE was dissolved in 5 ml tetrahydrofuran and 189 mg (0 . 600 millimoles) of tetrabutyl ammonium fluoride was added and agitated until dissolved. The reactants were 10 let to stand over night at ro~m temperature (200C).
The solvent was evaporated under reduced pressure and the residue was dissolved in lO ml chloroform, washed with two successive 10 ml portions of water, and centri-fuged to separate chloroform and water phases. The 15 chloroform phase was e~,aporated under vacuum to obtain 390 mg of oran-3_ b~ o..~l wax, which was det~rm; ne~ to be impure N-polyethylene glycol 1500 PE compound.
The wax was re-dissolved in 5 ml chloroform an~1 trans-ferred to the top of a 21 X 270 mm column of si I ica gel 20 moistened with chloroform. The column was de reloped by passing 100 ml of solvent through the column~ rhe Ta~le 2 solvents were used in se~lu~nce:
Table 2 Volume % Volume % Methanol Cnnt~; n; ng Chloroform 2% Conc. ~nmonium Hydroxide/methanol 100% 0%
95% 596 90% 10%
85% 15%
80% 20%
70% 30%
60% 40%
50% 50%
0% 100%
= . ~
_ _ WO 91/05546 PCr/US
~ 206~178-Separated 50 ml fractions of column effluent were saved. ~he fractions of the column were separated by TLC
on Si-Cl8 reversed-phase plates. TLC plates were deve-loped with 4 volumes of ethanol mixed with l volume of water. visll~]; 7at~ on was done by exposure to iodine vapor .
, Only those fractions containing an iodine-absorbing lipid of R~ about 0.20 were combined and evaporated to dryness under vacuum and dried in high vacuum to constant weight. In this way 94 mg of waxy crystalline solid was obtained of ~.W. 2226. The proton N~ spectrum of this material dissolved in deuterochloroform showed the ex-pected peaks due to the phosphatidyl ethanolamine portion of the molecule, together with a few methylene protons attributable to polyethylene glycol. (Delta = 3.7).
Example 4 Preparation of REVs and MLVs A. Sized REVs A total of 15 llmoles of the selected lipid components, in the mole ratios indicated in the examples below, were dissolved in chloroform and dried as a thin film by rotary evaporation. Thls lipid f~ lm w~s ~ic-solved in l ml of diethyl ether washed with distil ed water. To this lipid solution was added 0.34 ml of an aqueous buffer solution c~ntA;n;ng 5 mM Tris, l00 mM
NaCl, 0.l mM EDTA, pH 7.4, and the mixture was emulsified by sonication for l minute, rr~nt~n;ng the temperature of the solution at or below room temperature. Where the liposomes were prepared to contain encapsulated ['25I]
tyraminyl-inulin, such was included in the phosphate buffer at a concentration of about 4 uCi/ml buffer.
The ether solvent was remoued under reduced pres-sure at room temperature, and the resulting gel was taken _ . ,~
WO 91/0~546 PCr/US90/06211 S
~,Q,6~
up in 0.1 ml of the above buffer, and shaken vigorously.
The res~ulting REV suspension had particle sizes, as determlned by microscopic examinatlon, of between about 0.1 to Z0 microns, and was composed pre~i~ ini~ntly of 5 relatively large ~greater than 1 micron) vesicles having one or only a few bilayer lamellae.
The liposomes were extruded twice through a poly-carbonate filter (Szoka, 1978), having a selected pore size of 0.4 microns or 0.2 mlcrons. Liposomes extruded through the 0.4 micron filter averaged 0.17+ (0.05) micron diameters, and through the 0.2 micron filter, 0.16 (0.05) micron diameters. Non-encapsulated [l'sI~ tyr-aminyl-inulin was removed by passing the extruded lipo-somes through Sephadex G-50 (Pharmacia).
B . Sized MLVs MUl~ r vesicle (MLV) liposomes were pre-pared according to standard procedures by dissolving a mixture of lipids in an organic solvent containing prima-20 rily CEICl~ and drying the lipids as a thin film by rota-tion under reduced pressure. In some cases a ra~ioactive label for the lipid phase was added to the lipic solution before drying. The lipid film was hydrated by a~.diticn of the desired aqueous phase and 3 mm glass beads fol-25 lowed by agitation with a vortex and shaking above thephase transition temperature of the phospholipid com-ponent for at least l hour. In some cases a radioactive label for the aqueous phase was included in the buffer.
In some cases the hydrated lipid was repeatedly frozen 30 and thawed three times to provide for ease of the follow-ing extrusion step.
The size of the liposome samples was controlled by extrusion through defined pore polycarbonate filters using pressurized nitrogen gas. In one procedure, the .
WO9l/05546 PCr/U, 0 ~
20~717~8.
liposomes were P~tr~ od one time through a filter with pores of 0 . 4 ~m and then ten times through a filter with pores of 0.1 ~m. In another procedure, the liposomes were extruded three times through a filter with 0.2 ~m 5 pores followed by repeated extrusion with 0 . 05 llm pores until the mean diameter of the particles was oelow 100 nm as det~rm; n~ri by DLS . Unencapsulated aqueous components were removed by passing the extruded sample through a gel permeation column separating the liposomes in the void 10 volume from the small molecules in the included volume.
C. Loading 67Ga Into DF-Cr~rt~n;n~ Liposomes The protocol for preparation of Ga67-DF labeled 15 liposomes as adaE~ted from known procedures ~Gabizon, 1989). Briefly, liposomes were prepared with the ion rhf~l ~tor desferal mesylate encapsulated in the internal aqueous phase to bind irreversibly Ga transported through the bilayer by llyd~ohy~luinoline (oxine~.
D. Dynamic Light Scattering Liposome particle size distribution measurements were obtained by DLS using a NICOMP Model 200 ~ h -Brookhaven Instruments 8I-2030AT autocorrelator attached.
25 The instruments were operated according to the manufac-turer' s instructions . The NICOMP results were expressed as the mean diameter and standard deviation of a Gaussian distribution of vesicles by relative volume.
Example 5 Liposome Blood Lifetime Mea~uL, --ts A. Measuring Blood Circulation Time and Blood/-RES Ratios In, vivo studies of liposomes were performed in two different animal models: Swiss-Webster mice at 25g each and l~hnr~tnry ratg at 200-300g each. The 8tudies in mice involved tail vein ini ection of liposome samples at 1 I~M
5 rhn~rhnliriA/mouse followed by animal 5~rrif;r~ after a de~ined time and tissue removal for label quantitation by gamma counting. The weight and percent of the injected dose in each tissue were A~t~rmin~A The studies in rat8 involved establishment of a chronic catheter in a femoral vein for 10 removal of blood samples at defined times after injection of liposome samples in a catheter in the other ~emoral artery at 3-4 ~lM rhns~hnl iriA/rat. The percent of the injected. dose L~ ; n i ns in the blood at several time points up to 24 hours waS A~t~rminPrl, B. Time Course of Liposome ~t~ntinn in the Bl.~.d~LI
PEG-PE composed of methoxy PEG, le~m~l~r weight 1900 and l-palmitoyl-2-oleyl-PE (POPE) was prepared as in Example 2.
The PEG-POPE lipid was combined with and partially llydL~Lated egg PC (PHEPC) in a lipid:lipid mole ratio of about 0.1:2, and the lipid mixture was hydrated and extruded through a 0.1 micron polyr~rhnn~te membrane, as described in Example 4, to produce MLV ' s with averAge size about o .1 micron . The MLV
lipids included a small amount of r~i; nl ;Ihal ~d lipid marker l~C-cholesteryl oleate, and the ~nr~rslll ~tl~A marker 3H-in-ulin.
The liposome composition was injected and the percent initial in~ected dose in mice was Af~t~rm~nl~d as described in Example 4, at 1, 2, 3, :, and 24 after injection.
Both lipid and encap~ulated marker3 3howed greater than 10~ of original injected do~e after 24 hour3.
C. 24 ~Iour Blood Liposome Levels Studies to determine percent injected dose in the 10 blood, and blood/Rl:S ratios of a liposomal marker, 24 hours after intravenous liposome in~ection, were carried out as described above. Llposome fo7~ ti ons having the compositions shown at the left in Table 3 below were prepared as described above. Unless otherwise noted, the 15 lipid-derivatized PEG was PEG-l900, and the liposome size was 0 .1 micron . The percent dose ~ a - ~ n; ng in the blood 24 hours after intravenous administration, and 24-hour blood/F~ES ratios which were measured are shown in the center and right columns in the table, respectively.
Table 3 LiDid ~ s;t~nn~ 24 ~ours Arter IV Dose ~ n~ected Do-e in Bloo~ B/P~E:
PG:PC:Cho_ ( 75:9.25:5) ~. n.ol Pt: Chol (.0:5) ~ 3 P-G-DSPE:-C:Chol 2 .
30 P_G--DSPE: 'C:Chol (250 nm) I.n ~.~
P~.G""-D'PE:PC:Chol 2 .. 0 ~
P Gu -DS~'- :PC:Chol .
P G-)S~'F: 'C (0.75:9.25) 2 .C ~-~
P G-75-~E:PG:PC:Chol 4 ,.o 4.1) (~.7 i- .25:7:5) PEG-DS E:NaCholSO,:PC:Chol 25.0 2.5 (~7.7 :0.7S:9.25:4.25) ~'All fn l~tic~rq contain 33% rhnlpetprol and 7.5~ ch~rsed component and were 100 nm mean diameter except as noted. PEG-DSPE consisted Or PEG ,.c: excep~ as noted.
A
~ 52 206;7~78 As seen, percent dose Ll ;nin~ in the blood 24 hours after injection ranged between 5-40% for liposomeg r~mtA;n;n~
PEG-derivatized lipids. By co~trast, in both liposome 5 f~ t~nq lacking PEG-derivatized lipids, less than 1~ of liposome marker remained after 24 hours. Also as seen in Table 3, blood-RES ratios increased from 0.01-0.03 in cortrol lipo60mes to at least 0 . 2, and as high as 4 . 0 in liposomes ~mnt:~in;n~ PEG-derivatiZed liposomes.
C. Blood li~etime mea~uL~ s with polylactic acid derivatized PE.
Studies to ~t~rm;n~o percent injected dose in the blood at several times a~ter intravenous liposome injection were carried out as A~qcr;h~ above. MLV liposome f~ lnt;~r~
having the - 't;,n Polylactic Acid-PB:~SPC:Chol at either 2: 3 . 5 :1 or 1: 3 . 5 :1 weight % were prepared .
These data indicate that the ~ r~n~ ~ o~ the polylactic acid-coated liposomes is severalfold slower than similar t; l~nq without polylactic acid derivatized PE.
D. Blood lifetime meaYuL~ tq with polyglycolic acid Derivatized PE.
Studies to ~t.~rm; n~ percent injected dose ln the blood at several times a~ter intravenous liposome injection were carried out as described above. MI,V liposome fnrm ~ t; ~n having the composition Polyglycolic Acid-PE:~SPC:Chol at 2: 3 . 5 :1 weight % were ~repared.
' 53 ' 2067 t 78 These d~ita indicate that the clearance of the polyglycolic acid-coated liposomes is severalfold slower than similar formulations without polyglycolic acid deri~ratized PE.
r le 6 Rf~ect of Phnspho~ ;pid ~-yl-O'hA;n Sat~ration on Bloo~/TR~ ~Atios ;n PEG-pE T,;posr~m~
PEG-PE composed of methoxy PEG, molecular weight 1900 and distearylPE (DSPE) was prepared as in Example 2. The PEG-PE
lipids were f~ 1 A~ with selected lipids from among sphingomyelin (SM~, fully llydL~y~lated soy PC (PC), cholesterol (Chol), partially hydrogenated soy PC (PHSPC), and partially 1lydL~ ted PC lipids identiied as PC IVlr IV10, IV20, IV30, and IV40 in Table 4. The lipid components were mixed in the molar ratios shown at the left iII Table 5, and used to form MLV' g sized to 0 1 micron as described in Example 4 .
Table 4 Pb,i.~l. Trlm~ition Egg PC Te=p~r~ture Rang~ Mol~ 9i ~:ltty Acid Co Ip.
'. 18:0 ~ 18 .'i ~li~L 20:1-4 22:0 22:1-ll~tive ~0 12 30 15 0 3 0 s 20IV 40 <0 14 32 4 0 3 0 4 IV 30 c20-3~ 20 39 0 1 2 3 4 ' 54 ' 206 7 1 78 ~a~
bl~ RES B/RES 9~ R~TnA;~;n~
PEG-PE:SM:PC:Chol 0.2:1:1:1 19.23 6.58 2.92 49.23 5 PEG- PE: PE~SPC: Chol 0.15:1.85:1 20.54 7.17 2.86 55.14 PEG- PE: PC IV1: Chol 0.15:1.85:1 17.24 13.71 1.26 60.44 PEG-PE:PC IVl:ChOl (two animal~) 10 0.15:1.85:1 19.16 10.07 1.90 61.87 PEG - PE: PC IVl 0: Chol ( two animal _ ) 0.15:1.85:1 12.19 7.31 1.67 40.73 PEG-PE:PC IV10:Chol 0.15:1.85:1 2.4 3.5 0.69 12.85 15 PEG-PE:PC IV20:Chol 0.15:1.85:1 24.56 7.52 3.27 62.75 PEG- PE: PC IV2 0: Chol 0.15:1.85:1 5.2 5.7 0.91 22.1 PEG-PE:PC IV40:Chol 20 0.15:1.85:1 19.44 8.87 2.19 53.88 PEG- PE: PC IV: Chol 0.15:1.85:0.5 20.3 8.8 2.31 45.5 PEG-PE:EPC:Chol 0.15:1.85:1 15.3 9.6 1.59 45.9 24 hours after injection, the percent material injected (as measured by percent of l~C-cholesteryl oleate) L~ in;n~
the blood and in the liver (L) and spleen (S) were fl~ rmin~
and these values are shown in the two data columns at the lef t in Table 5. The blood and L+S (RES) values were used to 3 0 calculate a blood/RES value for each composition. The column at the right in Table 5 shows total amount of radloactivity recovered The two low total recovery values in the table indicate anomalous clearance behavior.
The results from the table ~ ~ ~te that the blood/RES
ratios are largely independent of the fluidity, or degree of saturation of the phospholipid components forming the , '55 2067l78 liposomes. In particular, there was no systematic change in blood/RES ratio observed among liposomes rnntA;n;nr largely saturated PC ~ tA (e.g., IV1 and IV10 PC's), largely unsaturated PC components (IV40), and intermediate-saturation components (e.g., IV20) .
In addition, a comparison of blood/RES ratios obtained using the relatively saturated PEG-DSPE compound and the relatively unsaturated PEG-POPE compound (Example 5) indicates that the degree of saturation of the derivatized lipid is itself not critical to the ability of the liposomes to evade uptake by the RES.
~Am~le 7 Rffect of rhnlesterol Anrl ~thnl~yl~ted l~hnlestProl nn Bloo~/R~ RAt;ns ;n PEG-PE Li~os~ ~
15 A. Efect of added cholesterol PEG-PE composed oi methoxy PEG, molecular weight 1900 and DSPE was prepared as described in Example 2. The PEG-PE lipids were formulated with selected lipids ~rom among qrh;, yclin (SM), fully hydrogenated soy PC (PC), and cholesterol (Chol), as indicated in the column at the left in Table 5 below. The three f~ lAt;nn~ shown in the table contain about 30, 15, and 0 mole percent cholesterol. Both REV's (0.3 micron slze) and MLV's (0.1 micron size) were prepared, substantially as in Example 4, with encapsulated tritium-labeled inulin.
The percent encapsulated inulin L~ ;n;nr in the blood 2 and 24 hours after administration, given at the right in Table 6 below, show no mea6urable effect of cholesterol, in the range 0-30 mole percent.
' 56 20671 78 ~a~
Iniect:ed Dose H-Inuli~ In Bl~
~ ~B~ ~ 24 HR.
'H Aclueous Label I~C - J.ipid ~abel (~eakage ) 1) SM:PC:Chol:PEG-DSPE
1: 1: 1: 0.2 _ _ _ _ _ _ _ _ _ _ 100 nm MLV 19 5 48 24 300 nm REV 23 15 67 20 2 ) SM: PC: Chol: PEG-DSPE
1: 1: o.s: 0.2 _ _ _ _ _ _ _ _ 300 NM rev 23 l5 71 17 3 ) SM: PC: PEG-DSPE
1: 1: 0.2 _ _ _ _ _ 100 nm MLV 19 6 58 24 300 nm REV 32 23 76 43 B. Effect of ethoxylated cholesterol Methoxy-ethyoxy-cholesterol was prepared by coupling methoxy ethanol to cholesterol via the trifluorosulfonate coupling method described in Section I. PEG-PE composed 25 of methoxy PEG, molecular weight 1900 and DSPE was prepared as described in Example 2. The PEG-PE lipids were formulated with selected lipid~ from among distearylPC ~DSPC), partially hydrogenated soy PC
(PHSPC), cholesterol, and ethoxylated cholesterol, as 3 o indicated at the right in Table 7 . The data ~3how that (a) ethoxylated cholesterol, in combination with PEG-PE, gives about the same degree of ~nhAnl t of lipo~ome lifetime in the blood as PEG-PE alone By itself, the ethoxylated cholesterol provides a moderate degree of f~nhA- of liposome lifetime, but substantially less than that provides by PEG-PE
~able 7 t;nn ~ rntected r~n~3e rn RlnnS
I~C-Chol-Oleate 2 HR . 2 4 HR .
10 HSPC:Chol:PEG-DSPE 55 9 1.85: 1: 0.15 HSPC:Chol:PEG-DSPE:PEGs-Chol 57 9 1.85: 0.85: 0.15: 0.15 HSPC: Chol: HPC: PEG5 - Chol 15 2 15 1.85: 0.85: 0.15: 0.15 HSPC: Chol :HPG 4 1.85: 1: 0.15 F le 8 Effect of t'hArged T~ id Cc onent~ on Blood/RT~ pAtios in PEG-PE L;po~h5n~fl PEG-PE composed of methoxy PEG, molecular weight 1900 and DSPE was prepared as de3cribed in Example 2.
The PEG-PE lipids were formulated with lipids selected from among egg PG (PG), partially hydrogenated egg PC
(PHEPC), and cholesterol (Chol), as indicated in the Figure 7 The two formulations shown in the figure c-~ntA;n,~ about 4.7 mole percent (triangles) or 14 mole percent (circles) PG The lipids were prepared as MLV's, sized to 0.1 micron as in Example 4.
The percent of injected liposome dose present 0 25, 1, 2, 4, and 24 hours after injection are plotted for both formulations in Figure 7. As seen, the percent PG
WO 91/05546 PCr/lJS90/06211 ~= 58 in the compo~aition had little or no effect on liposome retention in the bloodstream. The rate of loss of encap-sulated marker seen is also similar to that observed for similarly prepared liposomes containing no PG.
Example 9 Plasma Kinetics of PEG-Coated and Uncoated I.iposomes PEG-PE composed of methoxy PEG, molecular weight l900 and distearylPE (DSPE) was prepared as in Example 2.
lO The PEG-PE lipids were formulated with PHEPC, and choles-terol, in a mole ratio of 0 .15 ~ 5: l . A second lipid mixture cr~nt~ i ned the same lipids, but without PEG-PE .
Liposomes were prepared from the two lipid mixtures as described in Example 5, by lipid hydration in the pre-15 sence of desferal mesylate, followed by sizing to 0 . lmicron, and removal of non-entrapped desferal by gel filtration with subser~uent loading of '7Ga-oxine into the liposomes. The unencapsulated 67Ga was removed during passage through a Sephadex G-50 gel exclusion cloumn.
20 Both compositions ~-~r,nt~lned lO umoles/ml in 0.15 M NaCl, 0 . 5 mM des f eral .
The two liposome compositions ~0 . 4 ml) were in~ected IV in animals, as described in Example 6. At time 0.25, l, 3 or 5 and 24 hours after in jection, blood samp' es 25 were removed and assayed for amount inulin rr-~n~n~J in the blood, expressed as a percentage of the amount mea-sured; ~ t~1y after injection. The results are shown in Figure 9. As seen, the PEG-coated liposomes have a blood halflife of about ll hours, and nearly 3096 o: the 30 injected material is pre5ent in the blood after 24 hours.
By contrast, 1nroat~cl liposomes showed a halflife in the blood of less than l hour. At 24 hours, the amount of in~ected material w~s und~tectab1e.
. _ WO 91/05546 PCr/US90/06211 208"7`178 Example 10 PreparatiOn of Doxorubicin Liposomes Vesicle-forming lipids containing PEG-PE, PG, PHEPC, and cholesterol, in a mole ratio of 0 . 3: 0 . 3: 1. 4: 1 were 5 dissolved in chloroform to a final lipid concentration of 25 llmol phospholipid/ml. Alpha-tocopherol (c~-TC~ in free base form was added in chloroform:methanol (2:1) solution to a final mole ratio of 0 . 5% . The lipid solution was dried to a thin lipid film, then hydrated with a warm (60C~ solution of 125 mM ammonium sulfate containing 1 mM des~eral. Hydration was carried out with 1 ml of aqueous solution per 5011mole phospholipid. The lipid material was hydrated with 10 freeze/thaw cycles, using liquid nitrogen and a warm water bath.
Liposome sizing wa3 performed by extrusion through two Nuclepore polycarbonate membranes, 3 cycles through 0.2 microns filters, and ten cycles through 0.05 micron filters. The final liposome size was 100 nm. The sized liposomes were then dialyzed against 50-100 volumes of 596 20 glucose three times during a 24 hour period. A fourth cycle was carried out= against 5% glucose titered to pH
6.5-7.0 for 1 hour.
A solution of doxorubicin, 10 mg/ml in 0 . 9% NaCl, and 1 mM desferal, was prepared and mixed with an eqL~al 25 volume of the dialyzed liposome preparation. The con-centration of drug in the mixture was about 5 mg/ml drug 50 umoles/ml phospholipid. The mixture was ; ncl~hated for 1 hours at 60C in a water bath with shaking. Untrapped drug was removed by passage through a Dowex 50 WX ~'esin 30 packed in a small column. The column was centrifuged in a bench top centrifuge for 5 minutes to completely e] ute the liposome suspension. Sterilization of the mixture was by passage through a 0 . q5 micron membrane, and the liposomes were stored at 5C.
WO 91/0~546 PCr/US90/06211 Example 11 Plasma Kinetics of Free and Liposomal Doxorubicin PEG-PE composed of methoxy PEG, molecular weight 5 1900 and distearylPE ~DSPE) was prepared as in Example 2.
The PEG-PE lipids were formulated with hydrogenated soy bean PC (HSPC) and cholesterol, in a mole ratio of 0.15:1.85:1 ~PEG-Dox). A second lipid mixture c~ntA;n~1 hydrogenated phosphatldylinositol ~HPI), HSPC choleste-10 rol, in a mole ratio of 1:10:5 (HPI-Dox). Each Iipid f,~rT~ t;~n was used in preparing sized MLVs containlng an ammonium ion gradient, as in Example 10.
The liposomes were loaded with doxorubicin, by mixing with an equal volume of a doxorubicin solution, 10 15 mg/ml plus 1 m~ desferal, as in Example 15. The two compositions are indicated in Figure 11 and Table 7 below as PEG-DOX and HPI-DOX liposomes, respectively. A doxo-rubicin HCl solution ~the Tn~rket~d product, Free Dox) was obtained from the hospital pharmacy. Free DOX, PEG-Dox 20 and HPI-Dox were diluted to the same ron~ntration ~1.8 mg/ml) using unbuffered 5% glucose on the dzy of in~ec-tion. Dogs were randomized into three groups ~2 females, 1 male) and weighed. An 18 gauge Venflon IV cathetc~- was inserted in a superficial limb vein in each animal. Th~:
25 drug and liposome suspensions were injected by quic!c bolus ~15 seconds). Four ml bllod samples were before in~ection and at 5, 10, 15, 30, 45 min, 1, 2, 4, 6, 8, 10, 12, 24, 48 and 72 hours post in~ection. In the lipo-some grOups blood was also drawn after 96, 120, 144; and 30 168 hours. Plasma was separated from the formed elements of the whole blood by centrifugation and doxorubi cin con~ ~ntrations assayed by standard fluorescence tech-niques. ~he amount of doxorubicin L~ ;n;ng in the blood was expressed as a percentage of peak concentration of ~ = . . .
61 2067 ~ 78 labeled drug, measured immediately after injection. The results are plotted in Figure 10, which shows that both the PEG-DOX and HPI-DOX compositions give linear logarithmic plots (single-mode exponential), and free drug give a bimodel 5 exponential curve, as indicated in Table 8 below. The halflives of the two liposome formulations rl,~t.~rm;n~l from these curves are indicated in Table 8.
Also shown in Table 8 is the area under the curve (AUC) determined by integrating the plasma kinetic curve over the 72 10 hour test period. The AUC results indicate that the total availability of drug from PEG-DOX liposomes, for the 72 hours period following injection, was nearly twice that of HPI-DOX
liposomes. This is consistent with the approximately twofold greater halflife of the PEG-DOX liposomes. The ~'CL" entry in 15 Table 9 indicates ...
Table 8 F~ee ~OX HPI-l~QX PEG-DQX
Kinetic Pattern Bi-exp. Mono-exp. Mono-exp.
Peak Conc 20(mg/1) 0 . 4-2 .2 ~ . 3 -6 . 0 4 . 5-5 . 0 AUC
(mg/1) 7.1-10.0 73.9-97.5 132.9-329.9 tl/2 hr 1.9-3.3 11.1-12 0 19.6-45.5 CL (mg/hr) 0.6-0.9 1.1-1.6 1.3-2.2 F le 1~
Ti~suf~ Distrihl-t;nn o~ Dn~nnlhir;n ASl1hcu~nf~o11q T
PEG-liposomes loaded with doxorubicin were prepared as in Example 11 (PEG-DOX liposomes) . Free drug used was clinic 3 0 material obtained from the hospital pharmacy .
Two groups of twelve mice were in~ected subcutaneously with 1o6 ~-6456 tumor cells. After 14 days the '62 ~ 717~
tumors had grown to about 1 cm3 in size in the subr--tAnPo--~
space and the animal3 were in~ected IV (tail vein) with 10 mg/kg doxorubicin as free drug (group 1) or encapsulated in PEG
liposomes (group 2). At 4, 24, and 48 hours after drug injection, four animals in each group were sacrificed, and sections of tumor, heart, and mu6cle ti3sue were excised. Each tissue was weighed, then homogenized and extracted for determination of doxorubicin concentration using a standard florescence assay procedure (Gabizon, 1989). The total drug measured in each homogenate was expressed as ,ug drug per gram tissue .
The data for drug distribution in heart, muscle, and liver are plotted in Figures llA and llB for free and liposome-associated doxorubicin, respectively. In Figure llA it is seen that all three tissue types take up about the same amount of drug/g tissue, although initially the drug is taken up preferentially in the heart. By contrast, when entrapped in PEG-liposomes, the drug shows a strong selective 1rrAl;7At;on in the tumor, with reduced levels in heart and muscle tissue.
,~ Asci~P~ --Two groups of 15 mice were in~ected interperitoneally with 106 ;r-6456 lymphoma cells. The tumor was allowed to grow for one-two weeks at which time 5 ml of ascites fluid had accumulated. The mice were then injected IV with 10 mg/kg doxorubicin either in free drug form (group i) or entrapped in PEG liposomes as described in Example 11 (group 2) . Ascites fluid was withdrawn 3~rom threa animals in each group at 1, 4, 15, 24 and 48 hours post treatment. The ascites tumor was further fractionated into cellular and fluid components by centrifugation (15 min. 5000 rpm). Free and liposome-bound drug in the supernatant was flf~t~rn-;nl~fl by passing the fluid through a Dowex ~X resin, a3 above, to remove free drug. The 63 2067 ~ 78 doxorubicia concentrationS in the ascites fluid, tumo-cells, superna~ant, and resin-treated supe-natant we-e then determined, and from these values, ~g doxorubic~n/-gram tissue was calculated. The vAlues for total doxoribicin ~n~Pntration in the acites fluid (solid rl; ~1~), in the ~upernatant in liposome-a~sociated 5 fonn, (that i~, after renoval of free drug from the supernatant) (solid triangles), and in i~olated tumor cells (solid circles) are plotted in ~igure 12. As seen, the total doxorubicin in the ascites fluid in-creased steadily up to about 24 hours, then dropped slightly over the next 24 hours. ~qost of the doxorubicin in the tumor is in liposome-entrapped form, demonstrating that liposomes are able to extravasate into solid tumors in intact form.
In a similar experiment two groups of twelve mice were implanted IP with ' he J-6456 lymphoma and the tumor was allowed to establish as described above. Once the ascites tumor had reached about 5 ml, one group of ani-mals was in~ected wlth 10 mg/kg free doxorubicin and the other group with 10 mg/kg doxorubicin entrapped in P~:G
liposomes. At 4, 24 and 48 hours post treatment ascites fluid and blood samples were withdrawn from f~ur animals in each group and the animals were sacrificed. Sections of liver and heart tissue were excised from ~ach animal, homogenized and drug cnncPntration assayed as described 2g above. Plasma was separated from whole blood by centri-fugation and drug concPntration assayed as stated above.
DoxorubiCin concPntration in the ascites ~luid wa3 also measured. The results are presented in Table 9. Plasma and ascites fluid levels are expressed as llg doxoruL-icin per ml and liver and heart tissue values as llg doxoru-bicin per gram tissue. The standard deviations for each measurement is shown in parentheses. As shown, there is considerably more doxorubicin in plasma for the group receiving the drug in PE(~ liposome entrapped form at all time points Ascites tumor levels are also higher in the liposome group, particularly at the longer time points (24 and 48 hours); ~i These data confirm the selective delivery of the drùg to the tumor by the PEG liposomes.
Table 9 Plasma llg/ml ~SD) Hours Free PEG-DOX
4 0.9 (0.0~ 232.4 (95.7) 24 0.0 118.3 (6.7) 48 0.0 84.2 (20.3) Ascites Tumor (tumor & fluid) 4 0.3 (0.1) 3.8 (2.0) 24 0.1 (0.1) 23.0 (8.9) 48 0.4 (0.3) 29.1 (2.0) Liver llg/grams (SD) 4 8 .1 ( 1. 4 ) undetectable 24 6.2 (4.8) 9.8 (5.9) 48 6.1 ~3.6) 10.2 (0.1) Heart 45.7 ~3.4) 2-4 ~0.9) 24 2.5 ~0.3) 2.1 ~0.4) 48 1.5 (0.6) 2.3 (0.1) Tumor/Heart 40 . 0052 0 . 63 24 0 . 04 _ 10 . 9 48 0 . 266 12 . 6 Example 13 Tumcr Uptake of PEG T ~ ros~ ~ Compared with Conventional 40 T.~ no_ ~ .
Two groups of 6 mice were injected subcutaneously with 105-10' C-26 colon carcinoma cells and the tumor was allowed to grow in the s~hct~t~nPc-us space until it reached a size of 45 about 1 cm' (about two weeks following in~ection). Each ~ ~.
WO 91/05546 PCr~u.,, '~
.
6~67178 group of anlmals was then in jected with 0 . 5 mg of: either conventional liposomes (100 nm DSPC/Chol, l:1) or PEG lipo-somes ~100 nm DSPC/Chol/PEG-DSPE, 10:3:1) which had been loaded with radioactive gallium as described in Example 4.
5 Three mice from each group were s~r;f;ced at 2, 24 and 48 hours post treatment, the tumors excised and weighed and the amount of r~ O~Ct;Vity qn~n~lf~erl using a gamma counter.
The result9 are presented in the following table and are expre9sed as the percent of the injected dose per gram 10 tissue.
Table lD
PEG ~ONVENTIONAL RATIO IN
lS TUMOR~
Blood Liver Tumor Blood Liver Tumor 2hr 38.2 7.2 3.8 34.1 11.0 3.7 1.0 2024 hr 15.1 14.6 4.2 7.6 21.6 3.9 1.1 48 hr 5.5 13.8 3.5 1.2 25.0 1.7 2.1 25 AE S ~ as amount or PEG T'~ divided by amount of con-ventiona` liposomes l o~ in the tumor Example 14 Liposome Extravasation into Intact Tumors:
Direct Microscopic V~ C~ 7aT j t~T~
PEG-PE composed oi methoxy PEG, molecu~ ar weight 1900 and distearylPE (DSPE) was prepared as in Example 2.
The PEG-PE lipids were formulated with HSPC, and choles-terol, in a mole ratio of 0.15:1.85:1. PEG-liposomes were prepared to contain colloidal gold particles (H~g).
The resulting MLVs were sized by eXtrusion, as a'~ove, to an average 0.1 micron size. Non-entrapped material was removed by gel filtration. The final crlnc~ntration of liposomes in the suspension was about 10 ~Imol/ml.
-- _ _ = . . ~
. .
2~67 1 78 In a first study, a normal mouse was injected I~7 with 0 . 4 ml of the above liposome formulation . Twen~y four hours after injection, the animal was sacrificed, and sections of the liver removed fixed in a standard water-soluble plastic resin. Thick sections were cut with a microtome and the sections stained with a solution of silver nitrate according to instructions provided with the nIntense 2" System kit supplied by Jannsen Life Sciences, Inc. (Kingsbridge, Piscataway, N.J. ) . The sections were further stained with eosin and hemotoxylin.
Figure 13A is a photomicrograph of a typically liver section, showing smaller, irregularly shaped Kupfer cells, such as cells 20, among larger, more regular shaped hepatocytes, sucn as hepatocyes 22. The Kupfer cells show large cnnrPn~ations of intact lip~somes, seen as small, darkly stained bodies, such at 24 in Figure 13A. The hepatocyte~ are largely free of liposomes, as would be expected.
In a second study, a C-26 colon carcinoma (about 10' was implanted in a mouse liver. Fourteen days post implantation, the animal was in~ected IV with 0.5 mg of the above liposomes. Twenty four hours later, the al imal was sacrificed, and the liver was perfusec, embeded, sectioned, and stained as above . The sect ions wer~
p~m; nPd for a capillary-fed tumor region . One exemplary region is seen in Eigure 13B, which shows a capillary 26 feeding a region of carcinoma cells, - such as cells 28.
These cells have characteristic staining patterns, and often include darkly stained nuclii in various stages of mitosis. The capillary in the figure is lined b~- an endothelial barrier 30, and just below that, a basement membrane 32.
A
2~67 1 78 It can be seen in Figure 13B that liposomes, such as liposomes 34, are heavily c~ncPntrated in the tumor re-gion, ad~acent the capillary on the tumor side of the endothelial barrier and basement membrane, and many lipo-5 somes are also dispersed throughout the intercellularfluid surrounding the tumor cells.
Figure 13C shows another region of the liver tumor from the above animal. Liposomes are seen throughout the intercellular fluid bathing the carcinoma cells.
In a third study, C26 colon carcinoma cells were injected s~hc~tAn~ously into an animal, and allowed to grow in the animal for 28 days. Thereafter, the animal was in~ected IV with 0.5 mg of the above liposomes.
Twenty four hours later, the animal was sacrificed, and 15 the tumor mass was exc.~sed. After --;nn, tumor mass was secti~ne~ on a microtome and stained as above.
Figure 13D shows a region of the tumor cells, including a cell 36 in the center of the figure which is ln late stage mitosis. Small, darkly stained liposomes are seen 20 throughout the intercP~ l Ar fluid.
Example 15 Tumor Treatment Method Vesicle-forming lipids cnntAin;n~ PEG-PE, PG, PHEPC, 25 and cholesterol and ~-TC in a mole ratio of 0 . 3: 0 . 3 1.4: 1: 0.2 were dissolved in chloroform to a final lipid crnrPntration of 25 ~mol rhosrhol;pirl~ml. The lipid mix-ture was dried into a thin film under reduced pressure.
The film was hydrated with a sol~t; on of .125M amm~nium 30 sulfate to form MLVs. The MLV suspension was frozen in a dry ice acetone bath and thawed three times and size~ to 80-100 nm. An Amm~n;~-m ion gradient was created substan-tially as desc_ibed in Example 10. The liposomes were loaded with epirubicin, and free ~unbound drug~ removed A
.
-6a also as ~escribed in Example 10 for doxorubicin. Thè
final concentration of entrapped drug was about 50-100 llg drug/~mol lipid. Epirubicin HCl and doxorubicin HCL, the commercial products, were obtained from the hospital 5 pharmacy.
About 10' cells C-26 colon carcinoma cells were iniected subcutaneously into three groups of 35 mice.
The groups were subdivided into 5 7-animal subgroups.
For the tumor suppression experiment shown in Figure 10 14A each subgro~p was injected IV with 0.5 ml of eithe_ saline vehicle control ~open circles), 6 mgtkg epirubicin (open triangles), 6 mg/kg doxorubicin (filled circles), or the drug-loaded liposomes (PEG-DOX liposomes~ at two doses, 6mg/kg (filled triangles) and 12 mg/kg (open 15 s~uares) on days 1, 8 and 15 following tumor cell implan-tation. Each group was followed for 28 days. Tumor size was measured for each animal on days 5,7,12,14,17,21,24 and 28. The growth of the tumor in each ~ubyLuu~ (ex-pressed as the mean tumor size of the individual animals) 2 0 at each time point is plotted in Figure 14A .
With reference to this figure, neither ~ree doxoru-bicin nor free epirubicin at 6 mg/kg si~n;firAntly sup-pressed tumor growth compared with the sali~e control.
In contrast, PEG liposome entrapped epirubicin hoth doses 25 si~n; f; ~Ant ly suppresses tumor growth. With -espect to survival of the animals at 120 days followlng tumor lmplAntat; c-n, none of the animals in the saline, epiru-bicin or doxorubicin groups survived whereas 5 out of the seven and seven out of seven survived in the 6 ~g/kg 30 liposome epirubicin and 12 mg/kg 1 ipoc~ - epirubicin groups, respectively.
- ~ The results of delayed treatment experiments using the same tumor model are presented in Figure 14B and 14C.
The same number of animals were inoculated with the same _ _ _ _ _ _ _ _ . _ .. _ _ . .... . ..
number of tumor cells as described above. The treatme~,~
groups in Figures 14B and 14C consisted of sa~_ne (solid line), 6 mg/kg epiru~icin (filled triangles), 6 mg/kg free epirubicin plus empty PEG liposomes (open circles) 5 and two doses of epirubicin entrapped in PEG liposomes, 6 mg~kg (filled triangles) and 9 mg/kg (open squares~. In contrast to the results presented in Figure 14A, only two treatments were given in these experiments: days 3 and 10 for the results plotted in Figure 14B; and days 10 and 10 17 for the results plotted in FLgure 14C. Importantly, in the case of the PEG liposome entrapped drug, both delayed treatment schedules at both dose levels result in tumor regreSSiOn whereas the free drug and free drug plus empty liposome treatment ~roups show only a mo~est retar-~5 dation in the rate of t~mor growth.
Example 16 Tumor Treatment Method PEG-DOX liposomes were prepared as in Example 15 20 except that doxorubicin was loaded in the liposomes to a final level of 60-80 ug/umoles total lipid. ~ doxorubi-cin HCl solution to be used as the free drug control was obtained from a hospital pharmacy. A total of 30 mice were in~ected IP with l0' J-6456 lymphoma cells. The 25 animals were divided into three l0-animal group~;, each of which was in~ected IV with 0 . 4 ml of either saline vehi-cle, 10 mg/kg doxorubicin solution or the doxorubicin-loaded liposomes at l0 mg/kg. Each group was rollowed ~or l00 days for number of surviving animals. The per-30 cent survivors for each treatment group is plotted inFigure 15.
As can be seen, free drug ~filled circles) provided little improvement in survival over the saline group (filled squares). In the animals treated with doxorubi-WO 91/05546 PCr/VS90/06211 ~ ,6~
~ 70 cin loaded PÉG-liposomes (filled triangles), however, about 50% of the animals survived over 40 days, 20% over 70 days, and 1096 survived until the experiment was ter-minated at 10 0 days .
Example 17 Reduced Toxicity of PEG-Liposomes Solutions of free doxorubicin HCl, epirubicin HCl were obtained as above. PEG-liposome formulations con-10 taining either doxorubicin or epirubicin, at a drugconcentration of 70-90 ug compound/umole liposome lipid, were prepared as described in EXample 16. Conventional liposomes (no PEG-derivatized lipid) were loaded with doxorubicin to a drug concentration of 40 ug/umole lipid 15 using standard t~ hn~ 5.
Each of the five f~ t t ons was administered to 35 mice, at a dose between 10 and 40 mg drug/kg body weight, in 5 mg/kG in~ - c, with five receiving each dosage.
The maximum tolerated dose given in Table 11 below is 20 highest dose which did not cause death or dramatic weight loss in the injected animals within 14 days. As seen from the data, both DOX-liposomes and PEG-DOX liposomes more than doubled the tolerated dose of doxorubicin over the drug in free form, with the PEG-DOX liposomes giving 25 a slightly higher tolerated dose. A similar result was obtained for doses of tolerated epirubicin in free and -lipl~so~al ~
Table 1 1 Maximum Tolerated Dose of DXN
(mg~Kg in mice) S DX~ 10-12 DoX-Lip 25-30 PEG-DXN-Lip 25-35 ~;P I 10 P~G-EPI 20 Example 18 Tumor Treatment ~qethod Conventional doxorubicin liposomes (L-DOX) were pre-pared according to publlshed methods. Briefly, a mixture of eggPG, Egg, PC, cholesterol and a-TC in a mole ratio of 0.3: 1.4: 1: 0.2 was made in chlorsform. The solvent was removed under reduced presssure and the dry lipld film hydrated with a solution of 155 mN NaCl rnnt~;n~n~ 2-5 mg doxorubicin HCl. The resulting ~5LV preparation was down-sized by extrusion through a series of polycarbonate membranes to a final size of about 250 nm. The free (~nentrapped) drug was remoYed by passing the suspension over a bed of Dowex resin. The final doxorubicin con-centration was about 40 per umole lipid.
Three groups of 7 mice were inoculated subcutane~us-ly with 10' - 10' C-26 colon carcinoma cells as detailed in Example 15. The animals were divided into ~hree, 7-ani~al treatment groups, one of which receivd 0.5 ml of saline vehicle as a control. The other two groups were treated with doxorubicin either as a free drug solution or in the form of L-DOX liposom~es at a dose of 10 mg,'kg.
The tret .~ s were given on days 8, 15 and 22 after tumor cell inoc~ t; nn . Tumor size was measured on the days tre~rmPnts were given and day 2B. As shown in Figure 16, the free druy (filled circles) suppressed tumor growth to a modest extent compared with the saline control (;o~id line). The tumor in the L-Dox-treated group (filled triangles ) grew slightly faster than the ~ree-dru~-treated group and slightly more slowly than in the untreate~ group . These results l nrl~ ~Ate that the 5 anti-tumor activity o~ the L-DOX preparation is about the same, and certainly no better than the same- dose of free drug. This stands in marked contrast to the results presented in Example 15 (and Figures 14A-C) which ~how that at comparable doses epirubicin entrapped in PEG-lO liposomes has dramatically better anti-tumor activity than ree dn~g 1D tbis Jame tumo:: model.
.
!
'~ .
A~
The lnvention includes, in one aspect, a liposome composition for use in localizing a compound in a solid tumor, as defined in Section IV below, via the blood-30 stream comprising: The liposomes forming the composition(i) are composed of vesicle-forming lipids and between l-20 mole percent of an vesicle-forming lipid derivatized with a hydrophilic polymer, and (ii) have an average size in a selected size range between about 0 . 07-0 .12 microns .
-The compound i5 con-ained in the liposomes in en,-apped form li.e., associated with the liposome membrane o~
encapsulated within the internal aqueous compartment of the liposome). In this context, vesicle-forming lipid is defiAed as any lipid that by itself or in comb nation with other lipids forms bilayer structures.
In a preferred embodiment, the hydrophilic polymer is polyethyleneglycol or poly lactic poly glycolic acid havir.~ a molecular weight between about 1, 000-5, OQ0 daltons, and is derivatized to a phospholipid.
For u~e in tumor treatment, the compound in one embodiment is im anthracycline antibiotic or plant alka--loid, at least about 80% of the ~ ?ou.,~ is in liposome-entrapped form, and the drug is present in the liposomes at a concentration of a~ least about 20 llg compound/umole liposome lipid in the case of the anthracycline antibio-tics and and 1 ug/umoles lipi:l in the case of the plant alkaloids .
In a related aspect, the invention inclu~os a com-position of liposomes characterized by:
(a) liposomes composed of vesicle-forming lipids and between 1-20 mole percent of an vesicle-fo~ina lipid derivatized with a hydrophilic polymer, ~b) a blood lifetime, as measured by the percent of a liposomal marker present in the blood 24 hours after IV
administration which is several times greater than that of liposomes in the absence of the derivatized lipids;
(c) an average liposome size in a selected size range between about 0.07-0.12 microns, and (d) the compound in liposome-entrapped form.
Also disclosed is a method of preparing an agent for loc~l;7?(tion in a solid tumor, when the agent is adminis-tered by IV injection. In this case, following IV ad~.i-nistration the agent is carried through the bloodstream A
WO 91/05546 PCr/US90/062tl 20~71~8 in liposome-entrapped form with little leakage of the drug durlng the first 48 hours post injection. By virtue of the low rate of RES uptake during this period, the liposomes have the opportunity to distribute to and enter 5 the tumor. Once within the interstitial spaces of the tumor, it is not necessary that the tumor cells actually internalize the liposomes. The entrapped agent is re-leased from the liposome in close proximity to the tumor calls over a period of days to weeks and is free to 10 further Penetrate into the tumor mass ~by a process of diffusion) and enter tumor cells directly - exerting its anti-proliferative activity. The method includes entrap-ping the agent in liposomes of the type characterized above. One liposome composition preferred for transpo=t-15 ing anthracycllne antibiotic or plant alkaloid anti-tumor agents to systemic solid ~ umors would contain high phase transition phospholipids and ~holesterol as this type of liposome does not tend to rel~ase these drugs while circ~ t ~ n~ through the bloodstream du i ~lg the f_rst 24-20 48 hours following administration.
In another aspect, the invention incl-~ldes a method for localizing a ~~ ~ou..d in a solid tumor ir a subject.
The method includes preparing a composition of liposo.nes ~i) composed of vesicle-forming lipids and between 1-20 25 mole percent of an vesicle-forming lipid derivatized with a hydrophilic polymer, (ii) having an average siz~ in a selected size range between about 0 . 07-0 .12 microns, and ~iii) c~nt~n~n~ the compound in liposome-entrapped form.
The compositi on is in jected IV in the sub~ect in an 30 amount sufficient to localize a therapeutically effective dose of the agent in the solid tumor.
These and other objects and features of the present invention will become more fully apparent when the fol-lowing detailed description of the invention is read in .
: -conjunc' ion with the accompanying drawings.
Brief Description of the Drawings Figure 1 illustrates a general reaction scheme for 5 derivatizing a vesicle-forming lipid amine with a polyal-kylether;
Figure 2 is a reaction scheme for preparing phospha-tidyleth~nolamine (PE) derivatized with polyethylene-qlycol via a cyanuric chloride linking agent;
Figure 3 illu$trates a reaction scheme for preparing phosphatidylethanolamine (PE) derivatized with polyethy-leneglycol by ~eans of a d;;mi~A~ole activating reagen~;
Figure 4 illustrates a reaction scheme for preparing phos~h~tidylethanolamine (PE) derivatized with polyethy-leneglycol by means of a trifluo,. ~hAn~o sulfonate reagent;
Figure 5 illustrates a v~sicle-forming lipid deriva-tized with polyethyleneglycol ~hrough a peptide (A), ester (8), and disulfide (C) linkag~;
Figure 6 illustrates a reaction sci;eme so- p-eparing phosphatidyle1-hAnol Am; n.o (PE) derivatiz~d with poly lactic acid or polyglycolic acid;
Figure 7 is a plot of liposome re.sidenc~ times in the blood, expressed in terms of percent injected dose as a function of hours after IV injection, for PEG-PE lipo-somes containing different amounts of ~hos~hAtidylglyce-rol;
Figure 8 is a plot slmilar to that of Figure 7, showing blood res; d~nce times of liposomes composed of pred: inAntly unsaturated phospholipid components;
Figure 9 is a plot similar to that of Figure 7, showing the blood residence times of PEG-coated liposomes (solid triangles) and conventional, uncoated liposomes ~ solid circles );
A
Figure 10 i8 a plot showing the kinetics of ., doxorubicin clearance from the blood of beagle dogs, for -drug administered IV in free form (open circles), in liposomes formulated with saturated phospholipids and llydLoy~llated phosphatidylinositol (HPI) (open squares), and in liposomes coated with PEG (open triangles);
Fiqures lL~ and llB are plots of the time course of doxorubicin uptake from the bloodstream by heart (solid ~l;i '-), muscle (solid circles), and tumor (solid triangles) for drug administered IV in free llA and PEG-1 ;ro~ 1 (llB) form;
Figure 12 is a plot of the time course of uptake of doxorubicin from the bluod~L~al,l by J-6456 tumor cells implanted interperitoneally (IP) in mice, as measured as total drug (filled ,i;; '-) as drug associated with tumor cells (solid circles) and liposome-associated form (solid triangles);
Figures 13A-13D are light mi~Loyl~plls showing localization of liposomes (small dark stained particles) in Kupfer cells in normal liver (13A), in the interstitial fluid of a C-26 colon carcinoma implanted in liver in the region of a capillary supplying the tumor cells (13B) and in the region of actively dividing C-26 tumor cells implanted in liver (13C) or subcutaneously (13D);
Figures 14A-14C are plots showing tumor size growth in days following subcutaneou6 implantation of a C-26 colon carcinoma, for mice treated with a saline control (open circles), doxorubicin at 6 mg/kg (filled circles), epirubicin at 6 mg/kg (open triangles), or PEG-liposome entrapped epirubicin at two do6es, 6 mg/kg (filled triangles) or 12 mg/kg (open squares) on days 1, 8 and 15 (14A); for mice treated with saline (solid line), 6 mg/kg epirubicin (closed circles), 6 mg/kg epirubicin plus empty liposomes, (open circles), or PEG liposome -9a- 206il 78 entrapped at two doses, 6 mg/kg (filled triangles) and 9 mg/kg topen squares) on days 3 and 10 (14B) or days 10 and 17 ( 14C);
Figure 15 i8 a plot 6howing percent survivors, in 5 days following interperitoneal implantation of a J-6456 lymphoma, for animals treated with doxorubicin in free form (c$osed circles) or PEG-liposomal form (solid triangles), or untreated animals (filled squares); and Figure 16 is a plot similar to that in Figure 14, 10 showing tumor size growth, in days following subcutaneous implantation of a C-26 colon carcinoma, for animals treated with a saline control (solid line), or animals treated with 10 mg/kg doxorubicin in free form (filled circles), or in conventional liposome~; (filled 15 triangles).
lO 2067 1 7~
Detailed Description of the Il~ven ion I. PreparatiOn of Derivatized Lipids Figure 1 shows a general reaction scheme f or prepa-5 ring a vesicle-forming lipid derivatized a biocompatible, hydrophilic polymer, as exemplified by polyethylene glycol (PEG), polylactic acid, and polyglycolic acid, all of which are readily water soluble, can be coupled to vesicle-forming lipids, and are tolerated in vivo without 10 toxic effects. The hydrophilic polymer which is em-ployed, e.g., PEG, is preferably capped by a methoxy,ethoxy or other unreactive group at one end or, alte-na-tively, has a chemical group that is more highly rea~~ive at one end than the other. The polymer is activated at A
WO 91/0~46 PCr/US90/06211 .
2~67178 one of its ends by reaction with a suitable activating agent, such as cyanuric acid, diimadozle, anhydride reagent, or the like, as described below. The activated compound is then reacted with a vesicle-~orming lipid, 5 such as a diacyl glycerol, including diacyl phosphogly-cerols, where the two hydrocarbon chains are typically between 14-22 carbon atoms in length and have Yarying degrees of saturation, to produce the derivatized lipid.
phosrll~tidylethanol-amine (PE) is an example of a phos-10 pholipid which is preferred for t~Lis purpose since itcontains a reactive amino group which is convenient for coupling to the activated polymers. Alternatively, the lipid group may be activated for reaction with the poly-mer, or the two groups may be ~oined in a concerted 15 coupling reaCtiOn, according to known coupling methods.
PEG capped at one end with a methoxy or ethoxy group can be obtained commercially in a variety of polymer sizes, e.g., 500-20,000 dalton molecular weights.
The vesicle-forming lipid is pr~rQrably one having 20 two hydrocarbon chains, typically acyl chai~s, and a polar head group. Included in this class are the phos-pholipids, such as rh~srh~tidylcholine (PC), PE, phos-rh~ acid (PA), phosphatidylinositol (PI), and sphin-gomyelin (SM), where the two hydrocarbon chains are 25 typically between about 14-22 carbon atoms in length, and have varying degrees of unsaturation. Also included in this class are the glycolipids, such as cerebrosides and g~n~l i os1 ~l~os .
Another vesicle-forming lipid which may be employed 30 is cholesterol and related sterols. In general, choles-terol may be less tightly anchored to a lipid bilayer membrane, particularly when derivatized with a high molecular weight polymers, such as polyalkylether, and therefore be less effectlve in promoting liDosome evasion 6 PCr/US90/06211 of the RES in the bloodstream.
More generally, and as defined herein, "vesicle-forming lipid" is intended to include any amphipathic lipid having hydrophobic and polar head group moieties, 5 and which (a) by itself can form spontaneously into bilayer vesicles in water, as exemplified by phospholi-pids, or (~) is stably incorporated into lipid bilayers in combination with phospholipids, with its hydrophobic moiet~ in contact with the interior, hydrophobic region 10 of the b_layer membrane, and its polar head group moiety oriented toward the e~terior, polar surface of the mem-brane. An example of a latter type of vesicle-forming lipid is cholesterol and cholesterol derivatives, such as cholesterol sulfate and cholesterol hemisuccinate.
According to one important feature of the invention, the vesicle-forming lipid may be a relatively fluid lipid, typically meaning tha' the lipid phase has a relatively low liquid to liq.~id-crystalline melting temperature, e.g., at or below room temperature, or 20 relatively rigid lipid, meaning tha. the lipid has a relatively high melting temperature, ~.g., up to 60C.
As a rule, the more rigid, i.e., saturated lipiîl.~, con-tribute to greater membrane rigidity in a lipid bilayer structure and also contribute to greater bilayer stabi-25 lity in serum. Other lipid components, such as choleste-rol, are also known to contribute to membrane rigidlty and stability in lipid bilayer structures. A long ch~in (e.g. C-18) saturated lipid plus cholesterol is one preferred composition for delivering anthracycline anti-30 biotic and plant alkaloids anti-tumor agents to solid tumors since these liposomes do not tend to release the drugs into the plasma as they circulate through the bloodstream and enter the tumor during the first 48 hours following injection. Phospholipids whose acyl chains WO 91/05546 PCr/US90/06211 .
13 ~
have a variety of degrees of saturation can be obtained commercially, or prepared according to puolished methods.
Figure 2 shows a reaction scheme ~or producing a PE-PEG lipid in which the PEG is derivatized to PE through a 5 cyanuric chloride group. Details of the reaction are provided in Example 1. ~riefly, methoxy-capped PEG is activated with~ cyanuric chloride ln the presence in sodium carbonate under conditions which produced the activated PEG compound shown in the figure. This mate-10 rial is ~urified to remove unreacted cyanuric acid. Theactivated PE5 compound is reacted with PE in the presence of triethyl amine to produce the desired PE-PEG compound shown in the figure. The yield is about 8-10% with respect to initial riuantities of PEG.
The method just described may be applied to a vari-ety of lipld amines, lnr11-Aing PE, cholesteryl amine, and glycolipids with sugar-amine g oups.
A second method of coupling a polyalkylether, such as capped PEG to a lipid amine is ' llustrated i~. Figure 20 3. Here the capped PEG is activated witn a ~arbonyl m~ 7Ole coupling reagent, to form ~he activated imidazole compound shown in Figure 3. RePction with a lipid amine, such as PE leads to PEG coupli~g to the lipid through an amide linkage, as illustrated in the 25 PEG-PE compound shown in the figure. Details of the reaction are given in Example 2.
A third reaction method for coupling a capped poly-alkylether to a lipid amine is shown in Figure 4. Here PEG is ~irst protected at its OH end by a trimethylsilane 30 group. The end-protection reaction is shown in the figure, and involves the reaction of trimethylsilylchlo-ride with PEG in the presence of triethylamine. The protected PEG is then reacted with the anhydride of trifluoromethyl sulfonate to form the PEG compound acti-= =, , .
WO 91/05546 PCr/US90/06211 =-G~ 14 vated with trifluoromethyl sulfonate. Reaction of the activated compound wlth a lipid amine, such as PE, in the presence of = trie~hylamine, gives the desired derivatized lipid product, such as the PEG-PE compound, in which the 5 lipid amine group is coupled to the polyether through the terminal methylene carbon in the polyether polymer. The trimethylsilyl protective group can be released by acid treatment, as indicated in the figure, or, alternatively, by reaction with a quaternary amine fluoride salt, such l0 as the fluoride salt of tetrabutylamine.
It will be appreciated that a variety of known coupling reactions, in addition to those ~ust described, are suitable for preparing vesicle-forming lipids deriva-tized with hydrophilic polymers such as PEG,. For e~am-15 ple, the sulfonate anhyd. ide coupling reagent illustratedin Figure 4 can be used to ~oin an activated polyalkyl-ether to the hydroxyl group of an amphipathic lipid, such as the 5'-OH of cholesterol. Other reactive lipid groups, such as an acid or ester lipid group may also be 20 used for coupling, according to known coupling methods.
For example, the acid group of phosphatidi- acid can be activated to form an active lipid anhydride, by reaction with a suitable anhydride, such as acetic ~nhydride, a~d the reactive lipid can then be ~oined to a protected 25 polyalkylamine by reaction in the presence of an isothio-cyanate reagent. - -In another embodiment, the derivatized lipid c~m-ponents are prepared to include a labile lipid-polymer linkage, such as a peptide, ester, or disulfide linkage, 30 which can be cleaved under selective physiological condi-tions, such as in the presence of peptidase or esterase enzymes or reducing agents such as glutathione present in the bloodstream. Figure 5 shows exemplary lipids which are linked through (A) peptide, (B), ester, and (C), disul'ide containing linkages. The pep~ide-linked com-pound can be prepared, for example, by first coupling ~a polyalkylether with the N-terminal amine of the t-i~ep-tide shown, e.g., via the reaction shown in Figure 3.
5 The peptide carboxyl g-oup can then be coupled to a lipid amine group through a carbodiimide coupling reAgent con-ventionat ly . The ester linked compound can be prep2red, for example, by coupling a lipid acid, such as phospha~i-dic ac~ d, to the terminal alcohol group of a polyalkyl-10 ether, u~ing alcohol via an anhydride coupling agent.Alternatively, a ~hort linkage fragment c~nt~;n~rlg an internal ester bond and suitable end groups, such as primary amine groups can be used to couple the polyalkyl-ether to the amphipathic lipid through amide or ca:bamate 15 linkages. Similarly, the linkage fragment may contain an internal disulfide linkage, for use in forming the com-pound shown at C in Figure 5. Polymers coupled to phos-pholipids via such reversible inkages are useful to provide high blood levels of liposom~s which cont~in them 20 for the first few hours post injection. After this period, plasma components cleave the :ev~rsible bonds releasing the polymers and the "unprotected" i ~osomes are rapidly taken up by the RES.
Figure 6 illustrates a method for derivatizlng 25 polylactic acid with PE. The polylactic acid is reacted, in the presence of PE, with dicyclohexylcarboimide (DCCI), as detailed in Example 4. Similarly, a vesicle-forming lipid derivatized with polyglycolic acid may be formed by reaction of polyglycolic acid or glycolic acid 30 with PE in the presence of a suitable coupling agent, such as DCCI, also as detailed in Example 4. The vesi-cle-forming lipids derivatized with either polylac.ic acid or polyglycolic acid form part of the inven~ion herein. Also forming part of the inventiOn are liposomes A
WO 91/05546 PCr/US90/06211 1~ ~
?,~6 containing these derlvatized Lipids, in a 1-20 moLe percent .
II. Preparation of Liposome Composition 5 A. ~ipid Components The lipid components used in forming the liposomes of the invention may be selected from a variety of vesi-cle-forming lipids, typically including phospholipids, sphing~lipids and sterols. As will be seen, one require-lO ment of the liposomes of the present invention is longblood circulation lifetime. It is therefore useful to establish a standardized measure of blood lifetime which can be used for evaluating the effect =of lipid components on blood halflife.
one method used for evAlu~t;n~ l;ros~ - circulation tlme in vivo measures the distribution of IV injected liposomes in t~e bloodstream and the primary organs of the RES at selected times after injection. In the stan-dardized model which is used herein, RE~S uptake is mea-20 sured by the ratio of total liposomes l~ the bloodstream to total liposomes in the liver and spleen, the principal organs of the RES. In practice, age and sex matched mice are in~ected IV through the tail vein with a radiolab~' sd liposome composition, and each time point i~ determined 25 by measuring total blood and combined liver and spleen radiolabel counts, as detailed in Example 5.
Since the liver and spleen account for nearly 100%
of the initial uptake of liposomes by the RES, the blood-/RES ratio ~ust described provides a good approximation 30 of the extent of uptake from the blood to the RE~ ln vivo. For example, a ratio of about l or greater indi-cates a pr~ m; nAn~`p of injected liposomes remaining in the bloodstream, and a ratio below about l, a predomi-nance of Iiposomes in the RES. For most of the lipid WO 91/05546 PCr/US90/06211 compositions of interest, blood/RES ratios were calcu-lated at 1,2, 3, 4, and 24 hours post injection.
The liposomes of the present invention include 1-20 mole percent of the vesicle-forming lipid derivatized 5 with a hydrophilic polymer, described in Section I.
According to one aspect of the invention, it has been discovered that blood circulation halflives in these liposomes is largely independent of the degree of satura-tion of the phospholipid components making up the lipo-l0 somes. That is, the phospholipid components may becomposed of pr,orl~ ;n~ntly of fluidic, relatively unsatu-rated, acyl chains, or of more saturated, rigidifying acyl chain components. This feature of the invention is seen in Example 6, which Pl~m~ nPC blood/RES ratios in 15 liposomes formed with PE~-PE, cholesterol, and PC having varying degrees of saturation (Table 4)~. As seen from the data in Table 5 in the example, high blood/RES ratios were achieved in subst~nt;~1 1y ail of the liposome for-m111 at ~ t~nCi, t n~lprpn~pnt of the extenL of lipid ur,satura-20 tion in the bulk PC phospholipid, an~ no systematictrend, as a function of degree of lipid SaLuratiOn, waS
observed .
Accordingly, the vesicle-forming lipids may ~e selected to achieve a selected degree of fluidity or 25 rigidity, to control the stability of the liposomes in serum and the rate of release of entrapped drug from the liposomes in the bloodstream andtor tumor. The vesicle-forming lipids may also be selected, in lipid saturation characteristiCs, to achieve desired liposome preparation 30 properties. It is generally the case, for example, that more fluidic lipids are easier to formulate and down-size by extrusion and homogenization methods than more rigid lipid compositions.
:
, ~==
. '8 20671 78 Similarly, it h2s been found that the percen_age o~
cholesterol in the liposomes may be va_ied over a wi~e range wi~chout si~nifican~ effect on observed blood~REs ratios. The studies presenced in Example 7A, with refer-ence to Table 6 therein, show virtually no change in blood/RES ratios in the range of cholesterol between 0-30 mole percent.
~ It has also been found, in studies conducted in support o~ the invention, that blood~RES ratios are also relatively unaffected by the presence of charged lipid components, such as phos~hatidylglycerol tPG). This can be seen from Figure 7, which plots percent loss of encap-sulated marker for PEG-PE liposomes cont~in;ns either 4.7 mole percent PG ~triangles) or 14 mole percent P~; (cir-cles). Virtually no difference in liposome retention in the bloodstream over a 24 hour period was observed. The option of including negative charge in the liposome without aggravating RES uptake provides a number o~
potential advantages. Liposomes su~pensions which con-tain negative charge tend to be less s~nsi.ive to aggre-gation in high ionic strength buffers and hence physical stability is enhanced. Also, negative cha~go p~i~sent in the liposome membrane can be used as a formulat on ~ol to effectively bind high amounts of cationic drugs.
The vesicle-forming lipid derivatized wit~ a hydro-philic polymer is present in an amount preferably between about 1-20 mole percent, on the basis of moles of deriva-tized lipid as 2 percentage of total moles of vesicle-forming lipids. It will be appreciated that a lower mole ratio, such as less than 1. O mole percent, may be d}J~L~Liate for a lipid derivative with a large molecular weight polymer, such as one havlng a molecular weight o~ lOO kilodaltons. As noted in Section I, the hydrophilic polymer in the derivatized lipid preferably has a molecular weight WO 91/OSS46 - - PCr/l~S90/06211 ~ 20~7178 between about 200-20, 000 daltons, and more pre~erably between about 500-5, 000 daitons. Example ~B, which Am; nPC the effect of very short ethoxy ether moieties on blood/RES ratLos indlcates that polyether moLeties of 5 greater than about 5 carbon ethers are required to achieve signiflcant PnhAnc~=mPn~ of blood/RES ratlos.
B. Preparing the Liposome Composltion The liposomes may be prepared by a variety of tech-10 nlques, such as those detalled in Szoka et al, 1980. Onemethod for preparlng drug-containlng liposomes is the reYerse phase e~-aporation method described by Szoka et al and ln U.S. Patent No. 4,235,871. The reverse phase evaporatlon veslcles (REVs) have typical average slzes 15 between about 2-4 microl~iC and are preri( ~ nAntly oligo-l: ~ ~ l 1 Ar~ that 15~ coDtain one or a few lipid bilayer shells. The method is detaile~ in Example 4A.
~ lUlt~ 11 Ar vesicles (~V9) can be formed by simple lipid-film hydration techniques. In this proce-20 dure, a mixture of liposome-forming lipids of t he type detailed above dissolved in a suitable organ~ c solvent is evaporated in a vessel to form a thin fllm, whlch ls then covered by an aqueous medium, as detalled ln Example AB.
The lipid film hydrates to form ~Vs, typlcally with 25 sizes between about 0.1 to 10 microns.
In accordance with one important aspect of the invention, the liposomes are prepared to have suhstan-tially h~ ouS sizes in a selected slze range between about 0 . 07 and 0 .12 mlcrons . In partlcular, lt has been 30 discovered that liposomes in this size range are r~adily able to extravasate into solid tumors, as discussed in Sectlon III below, and at the same tlme, are capable of carrying a substantlal drug load to a tumor (unlike small ~ li -llAr vesicles, whlch are severely restricted in WO 91/05546 PCr/US90/06211 drug-loading capaclty). ~
One effecti~ve sizing method for ÆVs and MLVs in-volves extruding an aqueQus suspension of the liposomes through a series o~ = polycarbonate membranes having a selected uniform pore si2e in the range of 0. 03 to 0 .2 micron, typically 0.05, 0.08, 0.l, or 0.2 microns. The pore size of the membrane corresponds roughly to the largest sizes of liposomes produced by extrusion through that membrane, particularly where the preparation is l 0 extruded two or more times through the same membrane .
This method of liposome sizing is used in preparing homogeneous-si2e REV and ~qLV compositions described in the examples below. A more recent method involves extru-sion through an asymmetric ceramic i~ilter. The method is detalled in U.S. patent No. 4,,73;7,323 for Liposome Extru-sion issued April 12, 1988. Homogenization methods are also useful for down-si2ing li, osomes to sizes of l00nm or less (~artin).
C. ComE~ound Loading In one embodiment, the composition of ~e inventlon is used ~or loc~11 71n~ an imaglng agent, slch as radio-isotopes lncluding '7Ga and ~11In, or parama~netLc co.~-pounds at the tumor site. In this application, where the r~A1 O1 ahe1 can be detected at relatively low concentra-tion, it is generally sufficient to encapsulate the imaging agent by passive loading, i.e., during liposome formation. This may be done, for example, by hydrating lipids wlth an aqueous solutlon of the agent to be encap-sulated. Typically radiolabeled agents are radioisGtopic metals in ~h~ ted form, such as '7Ga-desferal, and are re~ained in the liposomes substantially ~ in entrapped form. After liposome formation and sl2lng, non-encapsu-lated material may be removed by one of a varlety of __ ~, , WO 9l/05j46 PCr/US9O/06211 ~ ~67178 methods, such as by ion exchange or gel filtration chro-matography. The concentration of chelated metal which can be achieved by passive loading is limited by the cnnr~n~ration of the agent in the hydrating medium.
Active loading of radioimaging agents is also pos-sible by entrapping a high affinity, water soluble chela-ting agent ~such as EDTA or desferoxamine) within the aqueous compartment of liposomes, removing any unen-trapped rhf~l at I nrJ agent by dialysis or gel exclusion column chromatography and incubating the liposomes in the presence of t~.e metal radioisotope chelated to a lower affinity, lipid ~oluble chelating agent such as 8-hydr-oxyriuinoline. The metal radioisotope is carried into the liposome by the lipid soluble chelating agent. Once in the liposome, the radiDisotope is chelated by the en-trapped, water soluble rhr~l ~t; ng agent - effectively trapping the radioisotope in the liposome interior (Gabi-zon) .
~assive loading may also b~ employed .'or the ~ ra~h~ c anti-tumor compounds, such as the alkaloids vinblastine and vincristine, which are the-~peutically active at relatively low drug doses, e.g., abou~ 1-15 mg/m2. Here the drug is either dLssolved in the ariueous phase used to hydrate the lipid or included with the lipids in liposome formation process, depending on the solubility of the compound. After liposome formation and slzing, free ~unbound) drug can be removed, as above, for example, by ion exchange or gel exclusion chromatographic methods .
Where the a~ti-tumor compound includes a peptide or protein drug, such as intrr1~1lkin-2 (IL-2) or tissue necrosis factor (TNF), or where the liposomes are formu-lated to contain a peptide immunomodulator, such as muramyl di- or tri-peptide derivatives or a protein WO 91/05546 PCr/US90/06211 ~ 6~ 22 immunomodulator such as macrophage colony _stimulating ~actor (M-CSF), the liposomes are preferably prepared by the above reverse phase method or by rehydrating a freeze dried mlxture of ~ t~e prptein and a sus~ension of small 5 unilamellar vesicles with water ~Kirby). Both methods combine passive loading with relatively high encapsu-lation efficiency, e.g., up to 50% efficiency. Nonencap-sulated materl~al can be readily removed ~rom the liposome suspension, e.g., by dialysis, diafiltratlon or exclusion lO chromatography.
~ he conc~ntr~t~nn o~ hydrophobic drug which can be accommodated in the liposomes will depend on drug/lipid interactions in the membrane, but is generally limited to a drug c~n- ~ntration of less than about 20 ~g drug/mg 15 lipid. More specifically, for a variety o~ anthracycline antibiotics, such as doxorubicin and epirubicin, the highest concentration of encapsulated material which can be achieved by passive loading intD the aqueo~s compart-ment of the liposome is about 10-20 Ils~umoles li}-id ~due 20 to the low intrinsic water solubi' ity of these compounds). When 20-30 mole percent of an a.iionic phos-pholipid such as PG is included in the membra}~e the loading factor can be increased to about ~ g/umole lipid because the anthracyclines are positively charged 2~ and form an "ion pair" complex with the negatively charged PG at the membrane interface. However, such charged complexed anthracycline form111at1nnc have limited utility in the context of the present invention ~which requires that the drug be carried through the bloodstream 30 ~or the first 24-48 hours following IV administration in liposome entrapped form) because the drugs tend to be rapidly released from the liposome membrane when intro-duced into plasma.
.
-In accordance with another aspect o~ the inventionl it has been found essential, for delivery of an therape,~-tically ef~ective dose of a variety of amphipathic anti-tumor drugs to tumors, to load the liposomes to a high drug concentration by active drug loading methods. For exat~ple, for anthracycline antibiotic drugs, such as doxorub~ cin, epirubicin, daunorubicin, carcinomycin, N-acetyladriamYCin, rubidazone, S-;mi~n~ nr ycin, and N-acetyldaunomycin, a final concentration o~ liposome-entrapped drug of greater than about 25 ~g/umole lipid and preferably 50 ~lg/umole lipid is desired. In~ernal drug cnn~ntrations as high as 100-200 ug/umole lipid are contemplated.
one method for active loading of amphipathic drugs into liposomes is described in co-owned U. S .
Patent ~o. ~,192,549. In this method, liposome~ are prepared in the pre~ence o E
a relatively high concentration of ~ ion, such as 0.125 ~ n sulfate~. After sizil:g the liposomes to a desired size, the llposome suspension is treated to create an inside-to-outside ammonium ion g. a~'ient across the liposomal membranes. The gradient may be crea.ed by dialysis against a non-ammonium ron~in;ng .nedi~m, such as an isotonic glucose medium, or by gel filtra~ion, such as on a Sephadex~ G-50 column equilibrated with 0.15~ NaCl or RCl, effectively replacing ammonium ions in th~ exte-rior phase with sodium or potassium ions. Alternat~;ely, the l 1ros suspension may be diluted with a non-am-monium solution, thereby reducing the exterior-phase cnn~ntration of ammonium ions. The ~ rn concen~ra-tion inside the liposomes is preferably at least 10 times, and more preferably at- least 100 to 1000 times that in the external liposome phase.
~Tradema~k ~,,.
- - -WO 91/05546 PCr1US90106~11 6t~
The ammonium ion graaient across the liposomes in turn creates a pH gradient, as ammonia is released across the liposome membrane,_and protons are trapped in the internal aqueous phase, of the liposome. To load lipo-somes wlth the selected drug a suspenslon of the lipo-somes, e.g., about 20-200 mg/ml lipid, is mixed with an aqueous solution of the drug, and the mixture is allowed to equilibrate over_ an period of time, e.g., several hours, at temperatures ranging from room temperature to 6~C - depending on the phase transition temperature of the lipids used to form the liposome. In one typical method, a suspension of liposomes having a lipid con-centration of 50 umoles/ml is mixed with an equal volume of anthracycline drug at a concentration of about 5-~
I5 mg/ml. At the end of the incubation period, the suspen-sion is treated to remove _ree (unbound) drug. One preferred method of drug removal for anthracycline drugs is by passage over an ion exchange resin, such ~s Dowex 50 WX-4, which is capable of binding ti~e drug.
Although, as noted above, the plant a ' kaloids such as vincristine do not require high loading factors in liposomes due to their intrinsically high anti-tumor activity, and thus can be loaded by passiv2 ~ntrapment techniques, it also possible to load these drug by active methods. Since vincristine is amphipathic and a weak base, it and similar molecules can be loaded into lipo-somes using a pH gradient formed by entrapping ammonium sulfate as described above for the anthracycline antibio-tics .
The remote loading method just described is il ` us-trated in Example l0, which descrlbes the preparation of 0.1 micron ~LVs loaded with doxorubicin, to a final drug concentration of about 80-lO0 ~Lg/umoles Iipid. The lipo-. _ = ~
WO 91/05546 PCr/US90/06211 .
20671'~{8 somes show a very low rate of drug leakage when stored at III. Liposome Localization in Solid Tumors A. ~rton~1~d Bloodstream Halflife One of the requirements for liposome localization in a target tumor, in accordance wlth the inventlon, is an exten~ed liposome lifetime~ ln the bloodstream following IV lipo-~ome administration. one measure of liposome lifetime in the bloodstream in the blood/RES ratio deter-mined at a selected time after liposome administration, as discussed above. Blood/RES ratios for- a variety of liposome compositions are given in Table 3 of Example 5.
In the absence of PEG-derivatized lipids, blood/RES
ratios were 0 . 03 or less . In the presence of PEG-deriva-tized iipids, the blood/RES ratio ranged from 0.2, for low-molecular weight PEG, to between l . 7-4 for several of the formulations, one of which lacks cholesterol, and three of which lack an added charged phospholipid (e.g., PG).
The data presented in Table 5 in Exa~ple 6 show blood/RES ratios ~excluding two points with low percent recovery) between about 1.26 and 3.27, cor si;t~nt with the data given in Table 3. As noted in Section II above, the blood lifetime values are subst~nt; ~1 1y independent of degree of saturation of the liposome lipids, presence of cholesterol and presence of charged lipids.
The blood/RES values reported above can be compared with blood/RES values reported in co-owned U . S . Patent No. 4, 920, 016, whiCh used blood/RES mea~uL~ -nt methods identical to those used in generating the data presented in Tables 3 and 5. The best 24-hour blood/RES ratios which were reported in the above-noted patent was 0 . 9, for a formulation composed of GMI, saturated PC, and 26 2~67 1 78 cholestero~. The next best formulations gave 24-hour blood/RES values of about 0 . 5 . Thus, typical 24-ho~ur blood/RES ratios ob-ained in a number of the current formulations were more than twice as high as the best 5 formulations which have been reported to date. Furthe-, ability to achieve high blood/RES with GMI or HPI lipids was dependent on the presence of prednr~i nAntly saturated lipids and cholesterol in the liposomes.
Plasma phArm~cokinetics of a liposomal marker in the lO bloodstream can provide another measure of the ~nhAnCPd liposome lifetime which is achieved by the liposome formulationS of the present invention. Figure~ 7 and 8 discussed above show the slow loss of liposomal marker from the bloodstream over a 24 hour period in typical 15 PEG-liposome form~1At;ons, substAnt jA1~y ~n~iPpen~Pnt of whether the marker is a lipid or an encArsul ~ted water-soluble compound lFigure 8). I;l both plots, the amount of liposomal marker present 24 ~ours after liposome injection is greater than 10% of the ~riginally injected 2 0 material .
Figure 9 shows the kinetics of liposom~ loss from the blood stream for a typical PEG-liposom0 form~:lation and the same liposomes in the absence of a ~r ' -deri~-a-tized lipid. ~fter 24 hours, the percent marker remain-25 ing in the PEG-liposomes was greater than about 2096, whereas the conventional liposomes showed less than 5%
retention in the blood after 3 hours, and virtuallY no detectable marker at 24 hours.
The results seen in Figures 7-9 are consistent with 30 24 hour blood liposome values measured for a variety of liposome formulations, and reported in Tables 3 and 5-7 in Example 5-8 below. As seen in Table 3 in Exam?le 5, the percent dose rPr~-;n;n~ at 24 hours was less than 1%
for conventional liposomes, versus at least 5% for -he A
PCI/US90/062tl WO 91/05~46 , , .
2067~7~ -PEG-liposomes. In the best form~ f 1 ons, values between about 20-40~6 were obtained. Similarly in Table 5 from - Example 6, liposome levels in the blood after 24 - hours (again neglecting two entries with low recovery values) 5 were between 12 and about 25 percent of total dose given.
Similar results are reported in Tables 6 and 7 of Example 7.
The ability of the liposomes to retain an amphi-pathic anti-tumor drug in the bloodstream over the 24-48 perlod required to provide an opportunity for the lipo-some to reach and enter a systemic tumor has also been investigated. In the study reported in Example ll, the plasma ~h~rm~sk~n~otics of doxorubicin loaded in PEG-liposomes, doxorubicin ~riven in free form, and doxorubi-cin loaded into liposomes contalning hydrogenated phos-phatidylinositol ~iPI) was in~ested in beagle dogs. The ~IPI liposomes were formulated wi'h a pre~ ~ n;lnt1y satu-rated PC lipid and cholesterol, and represents one of the optimal fQr~lAtion5 descr$bed in the above co-owr.ed U.S.
patent. The kinetics of doxorubicin in the blood up to 72 hours after drug administration is shown ir Figure ll.
Both liposomal fo lat~ons showed single-rrLode exponen-tial loss of drug, in contrast to free drug ~ h ~ 'i shows a bi-exp~n~ont ~ ~1 pattern . However, the amount of drug retained in the blood stream at 72 hours was about 8-10 times greater ln the PEG-liposomes.
For both blood~RES ratios, and liposome retention time in the bloodstream, the data obtained from a model animal system can be reasonably extrapolated to humans and veterinary animals of interest. This is because uptake of liposomes by liver and spleen has been f ound to occur at similar rates ln several mammalian species, including mouse, rat, monkey, and human (Gregoriadis, 1974; Jonah: Kimelberg, 1976, Juliano, Richardson;
. _ , ~
~ope~-Beresteir.). This result likely reflects the fact that the biochemical factors which appear to be m~st important in liposome uptake by the RES -- including opsinization by serum lipoproteins, size-dependent uptake 5 effects, and cell shielding by surface moieties -- are common features of all mammalian species which have been t'~Aml nf~d.
B. Extravasation into Tumors Another required feature for high-activity liposome targeting to a solid tumor, in accordance with the inven-tion, is liposome extravasation into the tumor through the endothelial cell barrier and underlying basement membrane separating a capillary from the tumor cells 15 supplied by the capillary. This feature is optimized in liposomes having sizes between 0 . 07 and 0 .12 microns .
That liposome delivery to the tumor is required for selective drug targeting can be seen from the study reported in Example 12. Here mice were inoculated sub-20 cutaneously with the J-6456 lymphoma whic~ formed a solid tumor mass of about 1 cm3 after one-two we~ks~ The ani-mals were then injected either with free dcxorubicin or doxorubicin loaded into PE&-liposomes at a :lo-s~ of l~m~
drug per kg body weight. The tissue distribution (heart, 25 muscle, and tumor) of the drug was then assayed at 4, 241 and 48 hours after drug administration. Figure llA shows the results obtained for free drug. No selective drug A~ m~ on into the tumor occurred, and in fact, the highest initial drug levels were in the heart, where 30 greateSt toxicity would be produced.
By contrast, drug delivery in PEG-liposomes showed increasing drug acc~m--l Ation into the tumor between 4-24 hours, and high selective tumor levels between 24 and 48 hours. Drug uptake by both heart and muscle tissue was, A
by contrast, lower than with free drug. As seen from the data plotted in Figure llB, the tumor cont~ined 8 ti~eS
more drug compared with healthy muscle and 6 times the amount in heart at 24 hours post injection.
To confirm that the PEG-liposomes deliver more anti-tumor drug to a intraperitoneal tumor, groups of mice were injected IP with 10~ J-64S6 lymphoma cells. After five Idays the IP tumor had been established, and the animals were treated IV with lOmg/kg doxorubicin, either in free drug form or entrapped in PEG-cont~;n;n~ lipo-somes. Tlssu~ distribution of the drug is tabulated in Table 9, Example 12. As shown, the tumor/heart ratio was about 272 greater for liposome delivery than for free drug at 24 hours, and about 47 times greater at 48 hours.
To demonstrate that the results shown in Table 9 are due to the entry of intact liposomes into the extravas-cular region of a tumor, the tu~or tissue was separated into cellular and s~rernPt~nt 'intercellular fluid) fractions, and the presence of liposome-associ.~ted and free drug in both fractions was assayed. Figure 12 shows the total amount of drug (filled ~ mr nr~ anc the amount of drug present in tumor cells (solid circle~) and in the supernatant in liposome-associated form (~olid triangles) over a 48-hour post injection period. To assay liposome-associated drug, the super-25 natant was passed through an ion-exchange resin to remove free drug, and the drug L. ~ I n~ ng in the supernatant was assayed (solid triangles). As seen, most of the drug in the tumor is liposome-associated.
Further demonstration of liposome extravasation into 30 tumor cells was obtained by direct microscopic observa-tion of liposome distribution in normal liver tissue and in solid tumors, as ~3~t~ led in Example 14. Figure 13A
shows the distribution of liposomes (small, darkly stained bodies) in normal liver tissue 24 hours after IV
injection of P~G-liposomes. The liposomes are confined exclusively to the KuDfer cells and are not prese~t either in hepatocytes or in the intercellular fluid ~f the normal liver tissue.
Figure 13B shows a region of C-26 colon carcinoma implanted in the liver of mice, 24 hours after injection of PEG-liposomes. Concentrations of liposomes are clear-ly evident in the region of the capillary in the figure, on the tumor tissue side of the endothelial barrier and basement membrane. Liposomes are also abundant in the intercellular fluid of the tumor cells, further eviden-cing passage from the capillary lumen into the tumor.
The Figure 13C photomicrograph shows another region of the tumor, where an abundance of liposomes in the inter-cellular fluid is also evident. A similar finding was made with liposome extravasation into a region of C-26 colon carcinoma cells injected sl~hcl~t~n~ously, as seen in Figure 13D.
IV. Tumor Localization ~ethod As detailed above, the liposomes of th~ invention are ef fective to localize specifically in a ~olid tumor region by virtue of the extended lifetime of ' he lipo-somes in the bloodstream and a liposome size which allows both extravasation into tumors, a relatively high drug carrying capacity and minimal leakage of the entrapped drug during the time required for the liposomes to dis-tribute to and enter the tumor (the first 24-48 hours following injection). The liposomes thus provide an effective method for loc~l;7;ng a compound selectively to a solid tumor, by entrapping the compound in such lipo-somes and injecting the liposomes ~V into a subject. In this context a solid tumor is defined as one that grows n ~n~omlc-l sl~e outslde the bloodstre~m (ln ~on-WO 91/0~546 PCr/US90/06211 2o67l78 trast, for example, to blood-born tumors such as leuke-mias) and requireS the formation of small blood vessels - - and capillaries to supply nutrients, etc. to the growing tumor mass. In this case, for an IV injected liposome - 5 (and its entrapped anti-tumor drug) to reach the tumor site it must leave the bloodstream and enter the tumor.
In one: -~;r L, the method is used for tumor treatment by lor~1l7in~r an anti-tumor drug selectively in the tumor. The anti-tumor drug which may be used is any compound, including the ones listed below, which can be stably entrapped in liposomes at a suitable loading factor and administered at a therapeutically effective dose (indicated below in parentheses after each compound) . These include ; h I r~h ~ c anti-tumor com-pounds such as the p~ ant alkaloids vincristine ~1. 4 mg/m2), vinblastine ~4-18 mg/m2) and etoposide (35-100 mg/m2), and the anthracycline antibiotics including doxo-rubicin (60-75 mg/m2), epirubicin (60-120 mg/m2) and daunorubicin (25-~5 mg/m2). The water-~soluble anti-meta-bolites such as methotrexate 3 mg/m2), c~tosine arabino-side (100 mg/m2), and fluorouracil (10-lS m3/kg), the antibiotics such as bleomycin (10-20 units/m2), mitomycin (20 mg/m2), plicamycin (25-30 ug/m2) and dactinc li~cin ;15 ug/m2), and the alkylating agents includlng cyclophospha-mide (3-25 mg/kg), thiotepa (0 . 3-0 . 4 mg/Kg) and BCNU
(150-200 mg/m2) are also useful in this context. Æs noted above, the plant alkaloids exemplified by vincris-tine and the anthracycline antibiotics including doxoru-bicin, daunorubicin and epirubicin are preferably active-ly loaded into liposomes, to achieve drug/lipid ratios which are several times greater than can be achieved with passive loading. Also as noted above, the liposomes may contain encapsulated tumor-therapeutic peptides and protein drugs, such as IL-2, andtor TNF, and/or immano-modulators, such as M-CSF, which are present alone or; in ccmbination with anti-tumor drugs, such as an anthraa~y-cline antibiotic drug.
The ability to ef~ectively treat solid tumors, in 5 accordance with the present invention, has been shown in a variety of in vivo systems. The method reported in Example 15 compares the rate of tumor growth in animals with ir~planted subcutaneously with a C-26 colon carci-noma. Treatment was with epirubicin, either in free 10 form, or entrapped in PEG-liposomes, in accordance with the invention, with the results shown in Figures 14A-C.
As seen, and discussed more fully in Example 15, treat-ment with epirubicin loaded PEG-liposomes produced a marked supression of tumor growth and lead to long term 15 survivors among groups of animals inoculated with a normally lethal dose of tumor cells. Moreover, delayed treatment of animals wlth the epiribicin loaded PEG lipo-somes resulted in regression of est~hl ~ ched subcutaneous tumors, a result not seen with free drug treatmen..
Similar results were obtained for treatment of a lymphoma implanted interperitoneally in mice, ~a~s detailed in Example 16. Here the animals were treate.~ with doxo-rubicin in free form or entrapped in P~3G- :- 70som~s .
Percent survivors over a 100-day period following tumor impl~n~isn and drug treatment is shown in Figure 16.
The results are similar to those obtained above, showing marked increase in the median survival time and percent survivors with PEG-liposomes over free drug treatment.
Since reduced toxicity has been observed in model animal systems and in a ~~lnic~l setting in tumor t3:eat-ment by doxorubicin entrapped in conventional liposomes ~as reported, for example, in U.S. Patent No. 4,898,735), it is of interest to determine the degree of toxicity protection provided in the tumor treatment method of the present invention. In the study reported ln Example 17, animals were injected Iv wlth increasing doses of doxo~ru-bicin or epirubicin in free form or entrapped in conven-tional or ~EG-liposomes, The maximum tolerated dose 5 ~TD) for the various drug formulations is given in Tzble lO in the Example. For both drugs, entrapment in PEG-liposomes appro~ t~l y doubled the ~TD of the drug .
Similar protection was achieved with conventional lipo-somes .
~th''U~Th reduced toxicity may contribute to the increased efficacy o~ tumor treatment reported above, selective lorll;7~ti~n of the drug by liposomal extrava-sation is also important for improved drug efficacy.
This is demonstrated in the drug treatment method de-15 scribed in Example 18. E~ere conventional liposomes cnnt~;n~ng doxorubicin (which show little or no tumor uptake by extravasation when administered IV) were com-pared with free drug at the sam~ dose (lO m~tkg) to reduce reduce the rate of growth of a subcuat i.neously 20 implanted tumor. Figure 16 plots tumor s~.ze with time in days following tumor implantation for a sal~ne control ~solid line), free drug (filled circles) and -o~ventional liposomes ~filled triangles) . As seen conver ti~nal l_po-somes do not supress tumor growth to any greater ~xtent 25 than free drug at the same dose. This finding stands in stark contrzst to the results shown in Figures 14A-C and 15 where improved survival and tumor growth supression is seen compared to free drug when tumor-bearing animals are treated wlth anthracycllnes anti-tumor drugs entrapped in 30 ~EG l; ros s .
Thus, the tumor-treatment method allows both higher levels of drug to be administered, due to reduced drug toxicity in liposomes, and greater drug efficacy, due to selective liposome localization in the intercellular A
WO 91/05546 Pcr/us9o .r fluid of the tumor.
It willj be appreciated that the ability to locali2e a compcund selectively in a tumor, by liposome extravasa-tion, can also be exploited for improved targeting of an 5 imaging agent to a tumor, for tumor diagnosis. Here the imaging agent, typically a radioisotope in chelated form, or a paramagnetic molecule is entrapped in liposomes, which are then administered IV to the sub ject being PxAm; nec~ . After a selected period, typically 24-48 10 hours, the subject is then monitored, for example by gamma scintillation radiography in the case of radioiso-tope or by N~qR in the case of the paramagnetic agent, to detect regions of local uptake of the imaging agent.
The following examples illustrate methods of 15 preparing liposomes with enhAn~P~ circulation times, and for accPssing circulation times in vivo and in vitro.
The examples are ~ntPn~led to illustrate srPr~f~c liposome compositions and methods of the inv~ntion, but are in no way intended to limit the scope thereof.
Materials Cholesterol (Chol) was obtained from Sigma (St.
Louis, NO) . Sphingomyelin (SN), egg phosphati~y] chol ne (lecithin or PC), partially hydrogenated PC havins the 2~ composition IV40, IV30, IV20, IV10, and IV1, phosphati-dylglycerol (PG), phnsph~tldylethanolamine (PE), dipalmi-toyl-phosphatidyl glycerol (DPPG), dipalmitoyl PC (DPPCl, dioleyl PC (DOPC) and distearoyl PC ~DSPC) were obta' ned from Avanti Polar Lipids (~irm~ngh~m, AL) or Austin 30 Chemical Company ~Chicago, IL).
["sI]-tyraminyl-inulin was made according to pub-lished procedures. 67Gallium-8-hyd,oxy~luinolLne was sup-plied by NEN Neoscan ~Boston, NA). Doxorubicin E~Cl and Epirubicin HCL were obtained from Adria Laboratorles (Colu;n~us. OH) or Farmitalia Carlo Erba (Mil2n, Italy) .
Example 1 Pre~aration of PEG-PE Linked by Cyanuric C h l o -5 ride A. Preparation of activated P~;G
2-0-Methoxypolyethylene qlycol 1900-4, 6-dichlo-ro-l,3,5 triazine previously called activated PEG was prepared as described in J. Biol. Chem., 252:3~82 ~1977) l0 with the following mo~fir~ons.
Cyanuric chloride (5.5 g; 0.03 mol) was dissolved in 400 ml of anhydrous benzene cont~;n;n~ 10 g of anhydrous sodium c~rh~"Ate, and PEG-1900 ~19 g; 0.01 mol) was added and the mixture was stirred overnight at room tempera-15 ture. The solution was ~iltered, and 600 ml of petroleumether (ho~ t ~ ng range, 35-60O) was added slowly with stir-ring. The ~inely divided precipitate was collected on a filter and redissolved in 400 ml o~ benzene. T~.e preci-pitation and ~iltration process was repeated several 20 times until the petroleum ether was free of residual cyanuric chloride as l~t~ n--d by high pres~-~re liquid chromatography on a column ~250 x 3.2 mm) of S-m "~i~hro-orb~" ~E. ~erck), developed with hexane, and det~ed with an ultraviolet detector. Titration of activated 25 PEG-1900 with silver nitrate after overni~ht hydrolysis in aqueous buffer at pH 10 . 0, room temperature, gave a value of 1. 7 mol of chloride liberated/mol of PEG.
T~C analysis of the product was effected with T~C
reversed-phase plates obtained from Baker using methanol-30 water, 4:1; v/v, as developer and exposure tO iodinevapor for vis~ Ation. Under these conditions; the startinq methoxy polyglycol 1900 appeared at R~=0.54 to 0 . 60 . The activated PEG appeared at Rf=0 . 41. Unreacted cyanuric chloride appeared at Rf=0 . 88 and was removed.
~Trademark A
WO 91/05~46 PCr/US90/06211 .~
The actlvated PEG was analyzed for nltrogen and an appropriate correctlon was applied ln selecting the quantity of reactant to use in further synthetic steps.
Thus, when t~he product contained only 20% of the theore-5 tical amoui~t of nitrogen, the quantity of material usedin the next synthetic step was increased by 10 0 /2 0, or 5-fold. When the product c-~nt~ 1 n~ 50% of the theore-tical amount of nltrogen, only 100/S0 or a 2-fold in-crease was needed.
B. Preparation of N- (4-Chloro-polyglycol 1900~-1,3,5-triazinyl egg phosphatldylethanolamine.
In a s~ ed test tube, 0.74 ml of a 100 mg/ml (0.100 mmole) stock solution of egg phosphatidylethanol-15 amine ln chloroform was evaporated to dryness under astream of nitrogen ana was added to the residue of the activated PEG described in secti on A, in the amount to provide 205 mg (0.100 mmole). To 1:his mixture, 5 ml an-hydrous dimethyl forTn~m1 rlP was added. 27 microliters 20 (0.200 mmole) triethylamine was added to ~he mixture, and the air was displaced with nitrogen gas. The ~.ixture was heated overnight in a sand bath maintained at 110C.
The mixture was then evaporated to cryn~ss ~ er vacuum and a pasty mass of crystalline solid was ob-25 tained. This solid was dissolved in 5 ml of a mixture of4 volumes of acetone and 1 volume of acetic acid. The resulting mixture was placed at the top of a 21 mm X 2gO
mm chromatographic absorption column packed with silica gel (~erck E~ieselgel 60, 70-230 mesh) which had first 30 been moistened with a solvent composed of acetone ac~tic acid, 80/20; v~v.
The column chromatography was developed with the same solvent mixture, and separate 20 to 50 ml aliquots of effluent were collected. Each portion of effluént was . _ .
.
WO 9l/05546 PCr/US90/06211 ~ 20871,7-~
2ssayed by lLC on silica gel coated plates, using 2-buta-none/acetic acid/water; 40/25/5; v/v/v as developer and iodine vapor exposure for visualization. Fractions containing only material of R~=about 0.79 were combined - 5 and evaporated to dryness under vacuum. Drying to con-stant weight under high vacuum afforded 86 mg (31. 2 micromoles) of nearly colorless solid N- ~4-chloro-poly-glycol 1900)-1,3,5-triazinyl egg phosphatidylethanolamine Cont 1 l n; n~ phosphorous .
The solid compound was taken up in 24 ml of etha-nol/chloroform; 50/50 chloroform and centrifuged to remove insoluble- material. Evaporation of the clarified solution to dryness under vacuum afforded 21 mg (7 . 62 micromoles) of colorless solid.
Example 2 Preparation of C~rh~m~te and Amide Linked Hydrophilic Polymers with PE
A. Preparation of the imidazole r~rh~m te cf poly-20 ethylene glycol methyl ether 1900.
9.5 grams (5 mmoles) of polyethylene gly;ol methylether l900 obtained from Aldrich Chemical Cc. was dis-solved in 45 ml benzene which has been drie ~ ov~r mo' e-cular sleves. 0.89 grams ~5.5 mmoles) of pure carbonyl 25 ~; im~rl~7ole was added. The purity was checked by an infra-red spectrum. The air in the reaction vessel was displaced with nitrogen. Vessel was enclosed and heated in a sand bath at 75C for 16 hours.
The reaction mixture was cooled and the clear solu-30 tion formed at room temperature. The solution was ~ilu-ted to 50 . 0 ml with dry benzene and stored in the refri-gerator as a 100 micromole/ml stock solution of the imidazole carbamate of PEG ether 1900.
WO 91/05546 PClr/US90/06211 ~S ~ ~ ~ _ ~ 6¢1 38 B. Preparation of the phosphatidylethanolamine car-bamate of polyethylene glycol methyl ether l900.
lO . 0 ml ~lmmol) of the lO0 mmol/ml stock solution of the imidazole carbamate of polyethylene glycol methyl ether l900 was pipetted lnto a lO ml pear-shaped flask.
The solvent was removed under vacuum. 3.7 ml of a lO0 mg/ml solutlon of egg phosphatidyl ethanolamine in chlo-roform (0.5 mmol) was added. The solvent was evaporated under vacuum. 2 ml of l, l, 2, 2-tetrachloroethylene and 139 microllters (l.0 mmol) of triethylamine VI was added.
The vessel was closed and heated in a sand bath main-tained at 95C for 6 hours. At this time, thin-layer chromatography was performed with fractions of the above mixture to determine an extent of con~ugation on Sl02 coated TLC plates, using butanone/acetic acid/water;
40/5/5; v/v/v; was performed as developer. I2 vapor V; Sl~A 1 i 7At ~ on revealed that most of the free phosphatidyl ethanolamine of Rf=0 . 68, had reacted, and was replaced by a phosphorous-c~ tA~n1n~ lipid at R~sO. ,8 to 0.80.
The solvent from the l~ ~ning reaction mixture was evaporated under vacuum. The residue was take.l up in lO
ml methylene chloride and placed at the top oE a 21 mm x 270 mm chromatographic absorption column pac;-~d w th ~erck Rieselgel 60 (70-230 mesh silica gel), which has been first rinsed with methylene chloride. The mixture was passed through the column, in sequence, using the following solvents.
-.
Table 1 Volume % of Volume % Methanol ml Methylene Chloride With 2% Acetic Acid 5~00 100%
200 95% 5%
200 90% 10%
200 85% 15%
200 60% 40%
50 ml portions of effluent were collected and each portion was assayed by TLC on SiO2 - coated plates, using 12 vapor absorption for v; c~ ; 7at inn after developmen~
with chloroform/methanol/water/c~nrPntrated ammonium hydroxide; 130/70/8/0.5%; v/v/v/v. Most of the phos-phates were found in fractions 11, 12, 13 and 14.
These fractions were ' ine~l, evaporated to dryness under vacuum and dried in high vacuum to constant weight.
They yielded 669 mg of colorless wax of phosphatidyl 20 etha-nolamine r~rhA~ e of polyethylene glycol methyl ether. This represented 263 ~icromoles and a vield of 52.6% based on the rhosE~h~t;~yl ethanolamine.
An N~R spectrum of the product dissol~-ed in deutero--chloroform showed peaks corresron~l; ng to the s~ctrum for 25 egg PE, together with a strong singlet due to the methy-lene groups of the ethylene oxide chain at Delta = ~ . 4 ppm. The ratio of methylene protons from the etr~ lene oxide to the t~rm;n~l methyl protons of the PE acyl - groups was large enough to confirm a molecular weight of 30 about 2000 for the polyethylene oxide portion of the molecule of the desired product polyethylene ~ col conjugated phosphatidyethanolamine c;~rh~ te, M.W. 2, 654 .
C. Preparation of polylactic acid amide of phosphotl-dyletanolamine .
A
,a_ WO 91/05546 PCr/US90/06211 __ - 40 200 mg (0.1 mmoles) poly (lactic acid), m. wt. s 2, 000 (ICN, Cleveland, Ohio) was dissolved in 2.0 ml dimethyl sulfoxide by heating while stirring to dissolve the material completely. Then the solutlon was cooled imme-diately to 65 C ~ and poured onto a mixture of 75 mg (0.1 mmoles) of distearylphosphatidyl-ethanolamine (cal.
Biochem, La Jolla) and 41 mg (0.2 mmoles) dicyclohexyl-carbodiimide. Then 28 ml (0.2 mmoles) of triethylamine was added, the air swept out of-the tube with nitrogen gas, the tube capped, and heated at 65C for 48 hours.
After this time, the tube was cooled to room tempera-ture, and 6 ml of chloroform added. The chloroform solution was washed with three s~lr~r~sc1~e 6 ml volumes of water, centrifuged after each wash, and the phases sepa-rated with a Pasteur pipette. The I` ; n; ng chloroform phase was filtered with suction to remove suspended distearolyph~srhAt ~ ~ylethanolamine . The filtrate was dried under vacuum to obtain 212 mq of semi-crystalline solid .
This solid was dissolved in 15 ml o~ a mixture of 4 volumes ethanol with 1 volume water and passed through a 50 mm deep and 21 mm diameter bed of H' Dowex c o cation exchange resin, and washed with 100 ml of the salr.e ~o,l-vent .
The filtrate was evaporated to dryness to obtain 131 mg colorless wax.
291 mg of such wax was dissolved in 2.5 ml chloroform and transferred to the top of a 21 mm x 280 mm colu.,ln of sLlica gel wetted with chloroform. The chromatogram was developed by passing through the column, in sequence~ 100 ml each of:
100% chloroform, 0% (1% NH,OH in methanol);
90% chloroform, 10% (1% NE~OH in methanol);
85% chloroform, 15% (1% NH~OH in methanol), , WO 91/05546 PCr/US90/06Zl I
2~67178 80% chloroform, 2Q9~; (196 NH,OH in methanol);
70% chloroform, 30% (1% NH~OH in methanol);
Individual 25 ml portions of effluent were saved and assayed by TLC on SFOz-coated plates, using CHCl3, CH,OH, H70, con. NH~OH, 130, 70, 8, 0.5 v/v as developer and I2 vapor absorption for visualization.
The 275-325 ml portions of column effluent contained a single material, PO, +, of R~ = 0 . 89 .
When c~ ' in~d and evaporated to dryness, these afforded 319 mg colorless wax.
Phosphate analysis agrees with a molecular weight of possibly 115, 000 .
Apparently, the polymerization of the poly (lactic acld) occurred at a rat~ comparable to that at which lt reac~ed with phosphatidylethanolamine.
This side-reaction could probably be minimized by working with more dilute solutions of the reactants, D. Preparation of poly (glycolic acid) amide of DSPE
~ mixture of 266 mg. ~3.50 mmoles) glycolic acid, 745 mg (3.60 mmoles) dicyclohexyl carbodiimide, 75 mg. (0.10 mmoles) distearoyl phosphatidyl eth~n~ ml n~, ~? mi~-o-liters (0.23 mmoles triethyl amine, and 5.0 ml dry ~im-ethyl sulfoxide was heated at 75 C, under a nitrogen atmosphere, cooled to room temperature, then diluted with an equal volume of chloroform, and then washed with three successive equal volumes of water to remove dim~thyl sulfoxide. Centrifuge and separate phases wit~ a Pasteur pipette each time.
Filter the chloroform phase with suction to remove a small amount of suspended material and vacuum evaporate the filtrate to dryness to obtain 572 mg. pale amber wax.
WO 9I/05546 PCr/US90/06211 6'~ ~ 42 Re-dissolve this material in 2 . 5 ml chloroform and transfer to the top of a 21 mm X 270 mm column of silica gel (Merck Hieselgel 60I which has been wetted with chloroform .
Develop the ~ hromatogram by passing through the column, in se~uence, 100 ml each of: ~
100% chloroform, 0 % tl% NH,OH in methanol);
90% chloroform, 1095 (1% NHIOH in methanol);
85% chloroform, 15% (1% NH~OH in methanol);
80% chloroform, 20% (1% NH~OH in methanol);
70% chloroform, 30% (1% NH~OH in methanol) .
Collect individual 25 ml portions of effluent and assay each by TLC on Si) 2-coated plates, using CH Cl3, CH3 OH, H20, con-NE~OH; 130, 70, 8, 0.5 v/v as developer.
Almost all the PO4 + material will be in the 275-300 ml portion of effluent. Evaporation of this to dryness under vacuum, followe~ by high-vacuum drying, affords 281 mg of colorless wax.
ph~srh~t,~ analysis suggests a molecular w6ight of 924, 000 .
Manipulation of solvent volume during re~ction and molar ratios of glycolic acid and dicyclohexyl carbodi-imide would probably result in other sized molecul es .
Example 3 Preparation of Ethylene-Linked PEG-PE
A. Preparation of I-trimethylsilyloxy-polyethylene glycol is illustrated in the reaction scheme sho~ in Figure 3.
15.0 gm tlO mmoles) of polyethylene glycol) M.Wt. 1500, (Aldrich Chemical) was dissolved in 80 ml benzene . I . 40 ml (11 mmoles) of chlorotrimethyl silane (Aldrich Chemi-cal Co. ) and 1.53 ml (lmmoles) of triethylamine was added. The mixture was stirred at room temperature under ' _ an inert atmosphere for 5 hours.
The mixture was filtered with suction to separate crystals of triethylammonium chloride and the crystals were washed with 5 ml benzene. Filtrate and benzene wash 5 liquids were . ~; ne~l . This solution was evaporated to dryness under vacuum to provide 15 . 83 grams of colorless oil which solidified on standing.
TLC of the product on Si-C1, reversed-phase plates using a mlxture of 4 volumes of ethanol with 1 volume of 10 water as developer, and iodine vapor visualization, revealed that all the polyglycol 1500 (Rt=0 . 93) has been consumed, and was replaced by a material of R~=0 . 82 . An infra-red spectrum revealed absorption peaks characteris-tic only of polyglycols.
Yield of I-trimethylsilyoxypolyethylene glycol, M.W.
1500 was nearly quantitative.
B. Preparation of trifluoromethane sulfonyl ester of ltrimethylsilyloxy-polyethylene glycol.
15.74 grams (10 mmol) of the crystalline I-trLmethyl-20 silyloxy polyethylene glycol obtained abov~ w~s dissolvedin 40 ml anhydrous benzene ar,d cooled in ~ bath of crushed ice. 1.53 ml (11 mmol) triethylamine and 1.85 ml (11 mmol) of trif~ rome~h~n~c~l fonic anhydride qbtained from Aldrich Chemical Co. were added and the mixture was 25 stirred over night under an inert atmosphere until the reaction mixture changed to a brown color.
The solvent was then evaporated under reduced pressure and the residual syrupy paste was diluted to lOû O ml with methylene chloride. Because of the great reactivity 30 of trifluo~o~ h~nP sulfonic esters, no further purif, ca-tion of the trifluoromethane sulfonyl ester of I-tri-methylsilyloxy polyethylene glycol was done.
C. Preparation of N-1-trimethylsilyloxy polyethylene glycol 1500 PE.
_ _ WO 91~05~46 PCr/US90/06211 .
lO ml of the methylene chloride=stock sQlution of the trifluoromethane sulfonyl ester of ;-trimethylsilyloxy polyet};Lylene glycol was evaporated i:Q dryness under - vacuum to obtain about 1.2 grams of residue ~approxi-- 5 mately 0.7 mmoies). To this residue, 3.72 ml of a ch~o-r~:form solution containlng 372 mg (0.5 mmoles) egg PE was added. To the resultLng solution, 139 microliters ~1. 0 mmole) of triethylamine was added and the solvent was evaporated under vacuum. To the obtained residue, 5 ml dry dimethyl f~ m~fl~ and 70 microliters (0.50 mmoles) - triethylamine (VI) was added. Air from the reaction vessel was displaced with nitrogen. The vessel was closed and heated in a sand bath a 110C for 22 hours.
The solvent was evaporated under vacuum to obtain 1.58 grams of brownish colored oil.
A 21 X 260 mm chromatographic absorption column filled with Kieselgel 60 silica 70-230 mesh, was p-epared and rinsed with a solvent composed of 40 volumes of butanone, ~5 volumes acetic acid and 5 volumes of water. The crude product was dissolved in 3 ml of the san~ s?lvent and transferred to the top ~of the chromatograph~ column. The chromatogram was developed with the same solvent and sequential 30 ml portions of effluent were assayed eac~
- ~-by TLC.
The TLC assay system used silica gel coated glass plates, with solvent combination butanone/acetic acid/wa-ter; 40/25/5; v/v/v. Iodine vapor absorption served for ~ v; su~ l i 7ation . In this solvent system, the N-l -tri-methylsilyloxy polyethylene glycol 1500 PE appeared at R,=0.78. Unchanged PE appeared at R~=0.68.
- The desired N-l-trimethylsilyloxy polyethylene glycol 1500 PE was a chief constituent of the~ 170-300 ml por-tions of column effluent. WhFn evapo~ated to dryness .
WO 91/05546 PCr/US90/06211 2~6717~
under vacuum these portions afforded 1~1 mg of pale yellow oil of compound. ~
D. Preparation of N-polyethylene glycyl 1500: phospha-tidyl-ethanolamine acetic acid deprotection.
Once-chromatographed, ~E compound was dissolved in 2 ml of tetrahydrofuran. To this, 6 ml acetic acid and 2 ml water was added. The resulting solution was let to stand for 3 days at 23C. The solvent from the reaction mix-ture was evaporated under vacuum and dried to constant weight to obtaln 75 mg of pale yellow wax. TLC on Si-C18 reversed-phase plates, developed with a mixture of 4 volumes ethanol, 1 volume water, indicated that some free PE and some polyglycol-like material formed during the hydrolysis.
The residue was dissolved in 0 . 5 ml tetrahydrofuran and diluted with 3 ml of a solution of ethanol water: 80:20;
v:v. The mixture was applied to the top of a 10 ~m X 250 mm chromatographic absorption column packed with octade-cyl bonded phase siLica gel and column was deve] oped with ethanol water 80:20% by volume, collecting s~quential 20 ml portions of effluent. The effluent was assayed by reversed phase TLC. Fractions cnntA1n~ng only pro~ct of Rf=0 . 08 to 0 .15 were combined. This was typically the 20-100 ml portion of effluent. When evaporated to dry-- ness, under vacuum, these portions afforded 33 mg of colorless wax PEG-PE corresponding to a yield of only 3%, - based on the starting phosphatidyl ethAnol~mi nP
NMR analysis indicated that the product incorporated both PE residues and polyethylene glycol residues, but that in spite of the favorable-appearing el - Al analy-sis, the chain length of the polyglycol chain has been reduced to about three to four et~llene oxide residues.
WO 91/05546 PCrJUS90/06211 .
c~'~'~ `
The product prepared was used for a preparation of PEG-PE
liposomes. ~; ~
., E. ~ Preparation of N-Polyethylene glycol 1500 P.E. by 5 fluorlde deprotection.
500 mg of crude N-1-trimethylsilyloxy polyethylene gly~ol PE was dissolved in 5 ml tetrahydrofuran and 189 mg (0 . 600 millimoles) of tetrabutyl ammonium fluoride was added and agitated until dissolved. The reactants were 10 let to stand over night at ro~m temperature (200C).
The solvent was evaporated under reduced pressure and the residue was dissolved in lO ml chloroform, washed with two successive 10 ml portions of water, and centri-fuged to separate chloroform and water phases. The 15 chloroform phase was e~,aporated under vacuum to obtain 390 mg of oran-3_ b~ o..~l wax, which was det~rm; ne~ to be impure N-polyethylene glycol 1500 PE compound.
The wax was re-dissolved in 5 ml chloroform an~1 trans-ferred to the top of a 21 X 270 mm column of si I ica gel 20 moistened with chloroform. The column was de reloped by passing 100 ml of solvent through the column~ rhe Ta~le 2 solvents were used in se~lu~nce:
Table 2 Volume % Volume % Methanol Cnnt~; n; ng Chloroform 2% Conc. ~nmonium Hydroxide/methanol 100% 0%
95% 596 90% 10%
85% 15%
80% 20%
70% 30%
60% 40%
50% 50%
0% 100%
= . ~
_ _ WO 91/05546 PCr/US
~ 206~178-Separated 50 ml fractions of column effluent were saved. ~he fractions of the column were separated by TLC
on Si-Cl8 reversed-phase plates. TLC plates were deve-loped with 4 volumes of ethanol mixed with l volume of water. visll~]; 7at~ on was done by exposure to iodine vapor .
, Only those fractions containing an iodine-absorbing lipid of R~ about 0.20 were combined and evaporated to dryness under vacuum and dried in high vacuum to constant weight. In this way 94 mg of waxy crystalline solid was obtained of ~.W. 2226. The proton N~ spectrum of this material dissolved in deuterochloroform showed the ex-pected peaks due to the phosphatidyl ethanolamine portion of the molecule, together with a few methylene protons attributable to polyethylene glycol. (Delta = 3.7).
Example 4 Preparation of REVs and MLVs A. Sized REVs A total of 15 llmoles of the selected lipid components, in the mole ratios indicated in the examples below, were dissolved in chloroform and dried as a thin film by rotary evaporation. Thls lipid f~ lm w~s ~ic-solved in l ml of diethyl ether washed with distil ed water. To this lipid solution was added 0.34 ml of an aqueous buffer solution c~ntA;n;ng 5 mM Tris, l00 mM
NaCl, 0.l mM EDTA, pH 7.4, and the mixture was emulsified by sonication for l minute, rr~nt~n;ng the temperature of the solution at or below room temperature. Where the liposomes were prepared to contain encapsulated ['25I]
tyraminyl-inulin, such was included in the phosphate buffer at a concentration of about 4 uCi/ml buffer.
The ether solvent was remoued under reduced pres-sure at room temperature, and the resulting gel was taken _ . ,~
WO 91/0~546 PCr/US90/06211 S
~,Q,6~
up in 0.1 ml of the above buffer, and shaken vigorously.
The res~ulting REV suspension had particle sizes, as determlned by microscopic examinatlon, of between about 0.1 to Z0 microns, and was composed pre~i~ ini~ntly of 5 relatively large ~greater than 1 micron) vesicles having one or only a few bilayer lamellae.
The liposomes were extruded twice through a poly-carbonate filter (Szoka, 1978), having a selected pore size of 0.4 microns or 0.2 mlcrons. Liposomes extruded through the 0.4 micron filter averaged 0.17+ (0.05) micron diameters, and through the 0.2 micron filter, 0.16 (0.05) micron diameters. Non-encapsulated [l'sI~ tyr-aminyl-inulin was removed by passing the extruded lipo-somes through Sephadex G-50 (Pharmacia).
B . Sized MLVs MUl~ r vesicle (MLV) liposomes were pre-pared according to standard procedures by dissolving a mixture of lipids in an organic solvent containing prima-20 rily CEICl~ and drying the lipids as a thin film by rota-tion under reduced pressure. In some cases a ra~ioactive label for the lipid phase was added to the lipic solution before drying. The lipid film was hydrated by a~.diticn of the desired aqueous phase and 3 mm glass beads fol-25 lowed by agitation with a vortex and shaking above thephase transition temperature of the phospholipid com-ponent for at least l hour. In some cases a radioactive label for the aqueous phase was included in the buffer.
In some cases the hydrated lipid was repeatedly frozen 30 and thawed three times to provide for ease of the follow-ing extrusion step.
The size of the liposome samples was controlled by extrusion through defined pore polycarbonate filters using pressurized nitrogen gas. In one procedure, the .
WO9l/05546 PCr/U, 0 ~
20~717~8.
liposomes were P~tr~ od one time through a filter with pores of 0 . 4 ~m and then ten times through a filter with pores of 0.1 ~m. In another procedure, the liposomes were extruded three times through a filter with 0.2 ~m 5 pores followed by repeated extrusion with 0 . 05 llm pores until the mean diameter of the particles was oelow 100 nm as det~rm; n~ri by DLS . Unencapsulated aqueous components were removed by passing the extruded sample through a gel permeation column separating the liposomes in the void 10 volume from the small molecules in the included volume.
C. Loading 67Ga Into DF-Cr~rt~n;n~ Liposomes The protocol for preparation of Ga67-DF labeled 15 liposomes as adaE~ted from known procedures ~Gabizon, 1989). Briefly, liposomes were prepared with the ion rhf~l ~tor desferal mesylate encapsulated in the internal aqueous phase to bind irreversibly Ga transported through the bilayer by llyd~ohy~luinoline (oxine~.
D. Dynamic Light Scattering Liposome particle size distribution measurements were obtained by DLS using a NICOMP Model 200 ~ h -Brookhaven Instruments 8I-2030AT autocorrelator attached.
25 The instruments were operated according to the manufac-turer' s instructions . The NICOMP results were expressed as the mean diameter and standard deviation of a Gaussian distribution of vesicles by relative volume.
Example 5 Liposome Blood Lifetime Mea~uL, --ts A. Measuring Blood Circulation Time and Blood/-RES Ratios In, vivo studies of liposomes were performed in two different animal models: Swiss-Webster mice at 25g each and l~hnr~tnry ratg at 200-300g each. The 8tudies in mice involved tail vein ini ection of liposome samples at 1 I~M
5 rhn~rhnliriA/mouse followed by animal 5~rrif;r~ after a de~ined time and tissue removal for label quantitation by gamma counting. The weight and percent of the injected dose in each tissue were A~t~rmin~A The studies in rat8 involved establishment of a chronic catheter in a femoral vein for 10 removal of blood samples at defined times after injection of liposome samples in a catheter in the other ~emoral artery at 3-4 ~lM rhns~hnl iriA/rat. The percent of the injected. dose L~ ; n i ns in the blood at several time points up to 24 hours waS A~t~rminPrl, B. Time Course of Liposome ~t~ntinn in the Bl.~.d~LI
PEG-PE composed of methoxy PEG, le~m~l~r weight 1900 and l-palmitoyl-2-oleyl-PE (POPE) was prepared as in Example 2.
The PEG-POPE lipid was combined with and partially llydL~Lated egg PC (PHEPC) in a lipid:lipid mole ratio of about 0.1:2, and the lipid mixture was hydrated and extruded through a 0.1 micron polyr~rhnn~te membrane, as described in Example 4, to produce MLV ' s with averAge size about o .1 micron . The MLV
lipids included a small amount of r~i; nl ;Ihal ~d lipid marker l~C-cholesteryl oleate, and the ~nr~rslll ~tl~A marker 3H-in-ulin.
The liposome composition was injected and the percent initial in~ected dose in mice was Af~t~rm~nl~d as described in Example 4, at 1, 2, 3, :, and 24 after injection.
Both lipid and encap~ulated marker3 3howed greater than 10~ of original injected do~e after 24 hour3.
C. 24 ~Iour Blood Liposome Levels Studies to determine percent injected dose in the 10 blood, and blood/Rl:S ratios of a liposomal marker, 24 hours after intravenous liposome in~ection, were carried out as described above. Llposome fo7~ ti ons having the compositions shown at the left in Table 3 below were prepared as described above. Unless otherwise noted, the 15 lipid-derivatized PEG was PEG-l900, and the liposome size was 0 .1 micron . The percent dose ~ a - ~ n; ng in the blood 24 hours after intravenous administration, and 24-hour blood/F~ES ratios which were measured are shown in the center and right columns in the table, respectively.
Table 3 LiDid ~ s;t~nn~ 24 ~ours Arter IV Dose ~ n~ected Do-e in Bloo~ B/P~E:
PG:PC:Cho_ ( 75:9.25:5) ~. n.ol Pt: Chol (.0:5) ~ 3 P-G-DSPE:-C:Chol 2 .
30 P_G--DSPE: 'C:Chol (250 nm) I.n ~.~
P~.G""-D'PE:PC:Chol 2 .. 0 ~
P Gu -DS~'- :PC:Chol .
P G-)S~'F: 'C (0.75:9.25) 2 .C ~-~
P G-75-~E:PG:PC:Chol 4 ,.o 4.1) (~.7 i- .25:7:5) PEG-DS E:NaCholSO,:PC:Chol 25.0 2.5 (~7.7 :0.7S:9.25:4.25) ~'All fn l~tic~rq contain 33% rhnlpetprol and 7.5~ ch~rsed component and were 100 nm mean diameter except as noted. PEG-DSPE consisted Or PEG ,.c: excep~ as noted.
A
~ 52 206;7~78 As seen, percent dose Ll ;nin~ in the blood 24 hours after injection ranged between 5-40% for liposomeg r~mtA;n;n~
PEG-derivatized lipids. By co~trast, in both liposome 5 f~ t~nq lacking PEG-derivatized lipids, less than 1~ of liposome marker remained after 24 hours. Also as seen in Table 3, blood-RES ratios increased from 0.01-0.03 in cortrol lipo60mes to at least 0 . 2, and as high as 4 . 0 in liposomes ~mnt:~in;n~ PEG-derivatiZed liposomes.
C. Blood li~etime mea~uL~ s with polylactic acid derivatized PE.
Studies to ~t~rm;n~o percent injected dose in the blood at several times a~ter intravenous liposome injection were carried out as A~qcr;h~ above. MLV liposome f~ lnt;~r~
having the - 't;,n Polylactic Acid-PB:~SPC:Chol at either 2: 3 . 5 :1 or 1: 3 . 5 :1 weight % were prepared .
These data indicate that the ~ r~n~ ~ o~ the polylactic acid-coated liposomes is severalfold slower than similar t; l~nq without polylactic acid derivatized PE.
D. Blood lifetime meaYuL~ tq with polyglycolic acid Derivatized PE.
Studies to ~t.~rm; n~ percent injected dose ln the blood at several times a~ter intravenous liposome injection were carried out as described above. MI,V liposome fnrm ~ t; ~n having the composition Polyglycolic Acid-PE:~SPC:Chol at 2: 3 . 5 :1 weight % were ~repared.
' 53 ' 2067 t 78 These d~ita indicate that the clearance of the polyglycolic acid-coated liposomes is severalfold slower than similar formulations without polyglycolic acid deri~ratized PE.
r le 6 Rf~ect of Phnspho~ ;pid ~-yl-O'hA;n Sat~ration on Bloo~/TR~ ~Atios ;n PEG-pE T,;posr~m~
PEG-PE composed of methoxy PEG, molecular weight 1900 and distearylPE (DSPE) was prepared as in Example 2. The PEG-PE
lipids were f~ 1 A~ with selected lipids from among sphingomyelin (SM~, fully llydL~y~lated soy PC (PC), cholesterol (Chol), partially hydrogenated soy PC (PHSPC), and partially 1lydL~ ted PC lipids identiied as PC IVlr IV10, IV20, IV30, and IV40 in Table 4. The lipid components were mixed in the molar ratios shown at the left iII Table 5, and used to form MLV' g sized to 0 1 micron as described in Example 4 .
Table 4 Pb,i.~l. Trlm~ition Egg PC Te=p~r~ture Rang~ Mol~ 9i ~:ltty Acid Co Ip.
'. 18:0 ~ 18 .'i ~li~L 20:1-4 22:0 22:1-ll~tive ~0 12 30 15 0 3 0 s 20IV 40 <0 14 32 4 0 3 0 4 IV 30 c20-3~ 20 39 0 1 2 3 4 ' 54 ' 206 7 1 78 ~a~
bl~ RES B/RES 9~ R~TnA;~;n~
PEG-PE:SM:PC:Chol 0.2:1:1:1 19.23 6.58 2.92 49.23 5 PEG- PE: PE~SPC: Chol 0.15:1.85:1 20.54 7.17 2.86 55.14 PEG- PE: PC IV1: Chol 0.15:1.85:1 17.24 13.71 1.26 60.44 PEG-PE:PC IVl:ChOl (two animal~) 10 0.15:1.85:1 19.16 10.07 1.90 61.87 PEG - PE: PC IVl 0: Chol ( two animal _ ) 0.15:1.85:1 12.19 7.31 1.67 40.73 PEG-PE:PC IV10:Chol 0.15:1.85:1 2.4 3.5 0.69 12.85 15 PEG-PE:PC IV20:Chol 0.15:1.85:1 24.56 7.52 3.27 62.75 PEG- PE: PC IV2 0: Chol 0.15:1.85:1 5.2 5.7 0.91 22.1 PEG-PE:PC IV40:Chol 20 0.15:1.85:1 19.44 8.87 2.19 53.88 PEG- PE: PC IV: Chol 0.15:1.85:0.5 20.3 8.8 2.31 45.5 PEG-PE:EPC:Chol 0.15:1.85:1 15.3 9.6 1.59 45.9 24 hours after injection, the percent material injected (as measured by percent of l~C-cholesteryl oleate) L~ in;n~
the blood and in the liver (L) and spleen (S) were fl~ rmin~
and these values are shown in the two data columns at the lef t in Table 5. The blood and L+S (RES) values were used to 3 0 calculate a blood/RES value for each composition. The column at the right in Table 5 shows total amount of radloactivity recovered The two low total recovery values in the table indicate anomalous clearance behavior.
The results from the table ~ ~ ~te that the blood/RES
ratios are largely independent of the fluidity, or degree of saturation of the phospholipid components forming the , '55 2067l78 liposomes. In particular, there was no systematic change in blood/RES ratio observed among liposomes rnntA;n;nr largely saturated PC ~ tA (e.g., IV1 and IV10 PC's), largely unsaturated PC components (IV40), and intermediate-saturation components (e.g., IV20) .
In addition, a comparison of blood/RES ratios obtained using the relatively saturated PEG-DSPE compound and the relatively unsaturated PEG-POPE compound (Example 5) indicates that the degree of saturation of the derivatized lipid is itself not critical to the ability of the liposomes to evade uptake by the RES.
~Am~le 7 Rffect of rhnlesterol Anrl ~thnl~yl~ted l~hnlestProl nn Bloo~/R~ RAt;ns ;n PEG-PE Li~os~ ~
15 A. Efect of added cholesterol PEG-PE composed oi methoxy PEG, molecular weight 1900 and DSPE was prepared as described in Example 2. The PEG-PE lipids were formulated with selected lipids ~rom among qrh;, yclin (SM), fully hydrogenated soy PC (PC), and cholesterol (Chol), as indicated in the column at the left in Table 5 below. The three f~ lAt;nn~ shown in the table contain about 30, 15, and 0 mole percent cholesterol. Both REV's (0.3 micron slze) and MLV's (0.1 micron size) were prepared, substantially as in Example 4, with encapsulated tritium-labeled inulin.
The percent encapsulated inulin L~ ;n;nr in the blood 2 and 24 hours after administration, given at the right in Table 6 below, show no mea6urable effect of cholesterol, in the range 0-30 mole percent.
' 56 20671 78 ~a~
Iniect:ed Dose H-Inuli~ In Bl~
~ ~B~ ~ 24 HR.
'H Aclueous Label I~C - J.ipid ~abel (~eakage ) 1) SM:PC:Chol:PEG-DSPE
1: 1: 1: 0.2 _ _ _ _ _ _ _ _ _ _ 100 nm MLV 19 5 48 24 300 nm REV 23 15 67 20 2 ) SM: PC: Chol: PEG-DSPE
1: 1: o.s: 0.2 _ _ _ _ _ _ _ _ 300 NM rev 23 l5 71 17 3 ) SM: PC: PEG-DSPE
1: 1: 0.2 _ _ _ _ _ 100 nm MLV 19 6 58 24 300 nm REV 32 23 76 43 B. Effect of ethoxylated cholesterol Methoxy-ethyoxy-cholesterol was prepared by coupling methoxy ethanol to cholesterol via the trifluorosulfonate coupling method described in Section I. PEG-PE composed 25 of methoxy PEG, molecular weight 1900 and DSPE was prepared as described in Example 2. The PEG-PE lipids were formulated with selected lipid~ from among distearylPC ~DSPC), partially hydrogenated soy PC
(PHSPC), cholesterol, and ethoxylated cholesterol, as 3 o indicated at the right in Table 7 . The data ~3how that (a) ethoxylated cholesterol, in combination with PEG-PE, gives about the same degree of ~nhAnl t of lipo~ome lifetime in the blood as PEG-PE alone By itself, the ethoxylated cholesterol provides a moderate degree of f~nhA- of liposome lifetime, but substantially less than that provides by PEG-PE
~able 7 t;nn ~ rntected r~n~3e rn RlnnS
I~C-Chol-Oleate 2 HR . 2 4 HR .
10 HSPC:Chol:PEG-DSPE 55 9 1.85: 1: 0.15 HSPC:Chol:PEG-DSPE:PEGs-Chol 57 9 1.85: 0.85: 0.15: 0.15 HSPC: Chol: HPC: PEG5 - Chol 15 2 15 1.85: 0.85: 0.15: 0.15 HSPC: Chol :HPG 4 1.85: 1: 0.15 F le 8 Effect of t'hArged T~ id Cc onent~ on Blood/RT~ pAtios in PEG-PE L;po~h5n~fl PEG-PE composed of methoxy PEG, molecular weight 1900 and DSPE was prepared as de3cribed in Example 2.
The PEG-PE lipids were formulated with lipids selected from among egg PG (PG), partially hydrogenated egg PC
(PHEPC), and cholesterol (Chol), as indicated in the Figure 7 The two formulations shown in the figure c-~ntA;n,~ about 4.7 mole percent (triangles) or 14 mole percent (circles) PG The lipids were prepared as MLV's, sized to 0.1 micron as in Example 4.
The percent of injected liposome dose present 0 25, 1, 2, 4, and 24 hours after injection are plotted for both formulations in Figure 7. As seen, the percent PG
WO 91/05546 PCr/lJS90/06211 ~= 58 in the compo~aition had little or no effect on liposome retention in the bloodstream. The rate of loss of encap-sulated marker seen is also similar to that observed for similarly prepared liposomes containing no PG.
Example 9 Plasma Kinetics of PEG-Coated and Uncoated I.iposomes PEG-PE composed of methoxy PEG, molecular weight l900 and distearylPE (DSPE) was prepared as in Example 2.
lO The PEG-PE lipids were formulated with PHEPC, and choles-terol, in a mole ratio of 0 .15 ~ 5: l . A second lipid mixture cr~nt~ i ned the same lipids, but without PEG-PE .
Liposomes were prepared from the two lipid mixtures as described in Example 5, by lipid hydration in the pre-15 sence of desferal mesylate, followed by sizing to 0 . lmicron, and removal of non-entrapped desferal by gel filtration with subser~uent loading of '7Ga-oxine into the liposomes. The unencapsulated 67Ga was removed during passage through a Sephadex G-50 gel exclusion cloumn.
20 Both compositions ~-~r,nt~lned lO umoles/ml in 0.15 M NaCl, 0 . 5 mM des f eral .
The two liposome compositions ~0 . 4 ml) were in~ected IV in animals, as described in Example 6. At time 0.25, l, 3 or 5 and 24 hours after in jection, blood samp' es 25 were removed and assayed for amount inulin rr-~n~n~J in the blood, expressed as a percentage of the amount mea-sured; ~ t~1y after injection. The results are shown in Figure 9. As seen, the PEG-coated liposomes have a blood halflife of about ll hours, and nearly 3096 o: the 30 injected material is pre5ent in the blood after 24 hours.
By contrast, 1nroat~cl liposomes showed a halflife in the blood of less than l hour. At 24 hours, the amount of in~ected material w~s und~tectab1e.
. _ WO 91/05546 PCr/US90/06211 208"7`178 Example 10 PreparatiOn of Doxorubicin Liposomes Vesicle-forming lipids containing PEG-PE, PG, PHEPC, and cholesterol, in a mole ratio of 0 . 3: 0 . 3: 1. 4: 1 were 5 dissolved in chloroform to a final lipid concentration of 25 llmol phospholipid/ml. Alpha-tocopherol (c~-TC~ in free base form was added in chloroform:methanol (2:1) solution to a final mole ratio of 0 . 5% . The lipid solution was dried to a thin lipid film, then hydrated with a warm (60C~ solution of 125 mM ammonium sulfate containing 1 mM des~eral. Hydration was carried out with 1 ml of aqueous solution per 5011mole phospholipid. The lipid material was hydrated with 10 freeze/thaw cycles, using liquid nitrogen and a warm water bath.
Liposome sizing wa3 performed by extrusion through two Nuclepore polycarbonate membranes, 3 cycles through 0.2 microns filters, and ten cycles through 0.05 micron filters. The final liposome size was 100 nm. The sized liposomes were then dialyzed against 50-100 volumes of 596 20 glucose three times during a 24 hour period. A fourth cycle was carried out= against 5% glucose titered to pH
6.5-7.0 for 1 hour.
A solution of doxorubicin, 10 mg/ml in 0 . 9% NaCl, and 1 mM desferal, was prepared and mixed with an eqL~al 25 volume of the dialyzed liposome preparation. The con-centration of drug in the mixture was about 5 mg/ml drug 50 umoles/ml phospholipid. The mixture was ; ncl~hated for 1 hours at 60C in a water bath with shaking. Untrapped drug was removed by passage through a Dowex 50 WX ~'esin 30 packed in a small column. The column was centrifuged in a bench top centrifuge for 5 minutes to completely e] ute the liposome suspension. Sterilization of the mixture was by passage through a 0 . q5 micron membrane, and the liposomes were stored at 5C.
WO 91/0~546 PCr/US90/06211 Example 11 Plasma Kinetics of Free and Liposomal Doxorubicin PEG-PE composed of methoxy PEG, molecular weight 5 1900 and distearylPE ~DSPE) was prepared as in Example 2.
The PEG-PE lipids were formulated with hydrogenated soy bean PC (HSPC) and cholesterol, in a mole ratio of 0.15:1.85:1 ~PEG-Dox). A second lipid mixture c~ntA;n~1 hydrogenated phosphatldylinositol ~HPI), HSPC choleste-10 rol, in a mole ratio of 1:10:5 (HPI-Dox). Each Iipid f,~rT~ t;~n was used in preparing sized MLVs containlng an ammonium ion gradient, as in Example 10.
The liposomes were loaded with doxorubicin, by mixing with an equal volume of a doxorubicin solution, 10 15 mg/ml plus 1 m~ desferal, as in Example 15. The two compositions are indicated in Figure 11 and Table 7 below as PEG-DOX and HPI-DOX liposomes, respectively. A doxo-rubicin HCl solution ~the Tn~rket~d product, Free Dox) was obtained from the hospital pharmacy. Free DOX, PEG-Dox 20 and HPI-Dox were diluted to the same ron~ntration ~1.8 mg/ml) using unbuffered 5% glucose on the dzy of in~ec-tion. Dogs were randomized into three groups ~2 females, 1 male) and weighed. An 18 gauge Venflon IV cathetc~- was inserted in a superficial limb vein in each animal. Th~:
25 drug and liposome suspensions were injected by quic!c bolus ~15 seconds). Four ml bllod samples were before in~ection and at 5, 10, 15, 30, 45 min, 1, 2, 4, 6, 8, 10, 12, 24, 48 and 72 hours post in~ection. In the lipo-some grOups blood was also drawn after 96, 120, 144; and 30 168 hours. Plasma was separated from the formed elements of the whole blood by centrifugation and doxorubi cin con~ ~ntrations assayed by standard fluorescence tech-niques. ~he amount of doxorubicin L~ ;n;ng in the blood was expressed as a percentage of peak concentration of ~ = . . .
61 2067 ~ 78 labeled drug, measured immediately after injection. The results are plotted in Figure 10, which shows that both the PEG-DOX and HPI-DOX compositions give linear logarithmic plots (single-mode exponential), and free drug give a bimodel 5 exponential curve, as indicated in Table 8 below. The halflives of the two liposome formulations rl,~t.~rm;n~l from these curves are indicated in Table 8.
Also shown in Table 8 is the area under the curve (AUC) determined by integrating the plasma kinetic curve over the 72 10 hour test period. The AUC results indicate that the total availability of drug from PEG-DOX liposomes, for the 72 hours period following injection, was nearly twice that of HPI-DOX
liposomes. This is consistent with the approximately twofold greater halflife of the PEG-DOX liposomes. The ~'CL" entry in 15 Table 9 indicates ...
Table 8 F~ee ~OX HPI-l~QX PEG-DQX
Kinetic Pattern Bi-exp. Mono-exp. Mono-exp.
Peak Conc 20(mg/1) 0 . 4-2 .2 ~ . 3 -6 . 0 4 . 5-5 . 0 AUC
(mg/1) 7.1-10.0 73.9-97.5 132.9-329.9 tl/2 hr 1.9-3.3 11.1-12 0 19.6-45.5 CL (mg/hr) 0.6-0.9 1.1-1.6 1.3-2.2 F le 1~
Ti~suf~ Distrihl-t;nn o~ Dn~nnlhir;n ASl1hcu~nf~o11q T
PEG-liposomes loaded with doxorubicin were prepared as in Example 11 (PEG-DOX liposomes) . Free drug used was clinic 3 0 material obtained from the hospital pharmacy .
Two groups of twelve mice were in~ected subcutaneously with 1o6 ~-6456 tumor cells. After 14 days the '62 ~ 717~
tumors had grown to about 1 cm3 in size in the subr--tAnPo--~
space and the animal3 were in~ected IV (tail vein) with 10 mg/kg doxorubicin as free drug (group 1) or encapsulated in PEG
liposomes (group 2). At 4, 24, and 48 hours after drug injection, four animals in each group were sacrificed, and sections of tumor, heart, and mu6cle ti3sue were excised. Each tissue was weighed, then homogenized and extracted for determination of doxorubicin concentration using a standard florescence assay procedure (Gabizon, 1989). The total drug measured in each homogenate was expressed as ,ug drug per gram tissue .
The data for drug distribution in heart, muscle, and liver are plotted in Figures llA and llB for free and liposome-associated doxorubicin, respectively. In Figure llA it is seen that all three tissue types take up about the same amount of drug/g tissue, although initially the drug is taken up preferentially in the heart. By contrast, when entrapped in PEG-liposomes, the drug shows a strong selective 1rrAl;7At;on in the tumor, with reduced levels in heart and muscle tissue.
,~ Asci~P~ --Two groups of 15 mice were in~ected interperitoneally with 106 ;r-6456 lymphoma cells. The tumor was allowed to grow for one-two weeks at which time 5 ml of ascites fluid had accumulated. The mice were then injected IV with 10 mg/kg doxorubicin either in free drug form (group i) or entrapped in PEG liposomes as described in Example 11 (group 2) . Ascites fluid was withdrawn 3~rom threa animals in each group at 1, 4, 15, 24 and 48 hours post treatment. The ascites tumor was further fractionated into cellular and fluid components by centrifugation (15 min. 5000 rpm). Free and liposome-bound drug in the supernatant was flf~t~rn-;nl~fl by passing the fluid through a Dowex ~X resin, a3 above, to remove free drug. The 63 2067 ~ 78 doxorubicia concentrationS in the ascites fluid, tumo-cells, superna~ant, and resin-treated supe-natant we-e then determined, and from these values, ~g doxorubic~n/-gram tissue was calculated. The vAlues for total doxoribicin ~n~Pntration in the acites fluid (solid rl; ~1~), in the ~upernatant in liposome-a~sociated 5 fonn, (that i~, after renoval of free drug from the supernatant) (solid triangles), and in i~olated tumor cells (solid circles) are plotted in ~igure 12. As seen, the total doxorubicin in the ascites fluid in-creased steadily up to about 24 hours, then dropped slightly over the next 24 hours. ~qost of the doxorubicin in the tumor is in liposome-entrapped form, demonstrating that liposomes are able to extravasate into solid tumors in intact form.
In a similar experiment two groups of twelve mice were implanted IP with ' he J-6456 lymphoma and the tumor was allowed to establish as described above. Once the ascites tumor had reached about 5 ml, one group of ani-mals was in~ected wlth 10 mg/kg free doxorubicin and the other group with 10 mg/kg doxorubicin entrapped in P~:G
liposomes. At 4, 24 and 48 hours post treatment ascites fluid and blood samples were withdrawn from f~ur animals in each group and the animals were sacrificed. Sections of liver and heart tissue were excised from ~ach animal, homogenized and drug cnncPntration assayed as described 2g above. Plasma was separated from whole blood by centri-fugation and drug concPntration assayed as stated above.
DoxorubiCin concPntration in the ascites ~luid wa3 also measured. The results are presented in Table 9. Plasma and ascites fluid levels are expressed as llg doxoruL-icin per ml and liver and heart tissue values as llg doxoru-bicin per gram tissue. The standard deviations for each measurement is shown in parentheses. As shown, there is considerably more doxorubicin in plasma for the group receiving the drug in PE(~ liposome entrapped form at all time points Ascites tumor levels are also higher in the liposome group, particularly at the longer time points (24 and 48 hours); ~i These data confirm the selective delivery of the drùg to the tumor by the PEG liposomes.
Table 9 Plasma llg/ml ~SD) Hours Free PEG-DOX
4 0.9 (0.0~ 232.4 (95.7) 24 0.0 118.3 (6.7) 48 0.0 84.2 (20.3) Ascites Tumor (tumor & fluid) 4 0.3 (0.1) 3.8 (2.0) 24 0.1 (0.1) 23.0 (8.9) 48 0.4 (0.3) 29.1 (2.0) Liver llg/grams (SD) 4 8 .1 ( 1. 4 ) undetectable 24 6.2 (4.8) 9.8 (5.9) 48 6.1 ~3.6) 10.2 (0.1) Heart 45.7 ~3.4) 2-4 ~0.9) 24 2.5 ~0.3) 2.1 ~0.4) 48 1.5 (0.6) 2.3 (0.1) Tumor/Heart 40 . 0052 0 . 63 24 0 . 04 _ 10 . 9 48 0 . 266 12 . 6 Example 13 Tumcr Uptake of PEG T ~ ros~ ~ Compared with Conventional 40 T.~ no_ ~ .
Two groups of 6 mice were injected subcutaneously with 105-10' C-26 colon carcinoma cells and the tumor was allowed to grow in the s~hct~t~nPc-us space until it reached a size of 45 about 1 cm' (about two weeks following in~ection). Each ~ ~.
WO 91/05546 PCr~u.,, '~
.
6~67178 group of anlmals was then in jected with 0 . 5 mg of: either conventional liposomes (100 nm DSPC/Chol, l:1) or PEG lipo-somes ~100 nm DSPC/Chol/PEG-DSPE, 10:3:1) which had been loaded with radioactive gallium as described in Example 4.
5 Three mice from each group were s~r;f;ced at 2, 24 and 48 hours post treatment, the tumors excised and weighed and the amount of r~ O~Ct;Vity qn~n~lf~erl using a gamma counter.
The result9 are presented in the following table and are expre9sed as the percent of the injected dose per gram 10 tissue.
Table lD
PEG ~ONVENTIONAL RATIO IN
lS TUMOR~
Blood Liver Tumor Blood Liver Tumor 2hr 38.2 7.2 3.8 34.1 11.0 3.7 1.0 2024 hr 15.1 14.6 4.2 7.6 21.6 3.9 1.1 48 hr 5.5 13.8 3.5 1.2 25.0 1.7 2.1 25 AE S ~ as amount or PEG T'~ divided by amount of con-ventiona` liposomes l o~ in the tumor Example 14 Liposome Extravasation into Intact Tumors:
Direct Microscopic V~ C~ 7aT j t~T~
PEG-PE composed oi methoxy PEG, molecu~ ar weight 1900 and distearylPE (DSPE) was prepared as in Example 2.
The PEG-PE lipids were formulated with HSPC, and choles-terol, in a mole ratio of 0.15:1.85:1. PEG-liposomes were prepared to contain colloidal gold particles (H~g).
The resulting MLVs were sized by eXtrusion, as a'~ove, to an average 0.1 micron size. Non-entrapped material was removed by gel filtration. The final crlnc~ntration of liposomes in the suspension was about 10 ~Imol/ml.
-- _ _ = . . ~
. .
2~67 1 78 In a first study, a normal mouse was injected I~7 with 0 . 4 ml of the above liposome formulation . Twen~y four hours after injection, the animal was sacrificed, and sections of the liver removed fixed in a standard water-soluble plastic resin. Thick sections were cut with a microtome and the sections stained with a solution of silver nitrate according to instructions provided with the nIntense 2" System kit supplied by Jannsen Life Sciences, Inc. (Kingsbridge, Piscataway, N.J. ) . The sections were further stained with eosin and hemotoxylin.
Figure 13A is a photomicrograph of a typically liver section, showing smaller, irregularly shaped Kupfer cells, such as cells 20, among larger, more regular shaped hepatocytes, sucn as hepatocyes 22. The Kupfer cells show large cnnrPn~ations of intact lip~somes, seen as small, darkly stained bodies, such at 24 in Figure 13A. The hepatocyte~ are largely free of liposomes, as would be expected.
In a second study, a C-26 colon carcinoma (about 10' was implanted in a mouse liver. Fourteen days post implantation, the animal was in~ected IV with 0.5 mg of the above liposomes. Twenty four hours later, the al imal was sacrificed, and the liver was perfusec, embeded, sectioned, and stained as above . The sect ions wer~
p~m; nPd for a capillary-fed tumor region . One exemplary region is seen in Eigure 13B, which shows a capillary 26 feeding a region of carcinoma cells, - such as cells 28.
These cells have characteristic staining patterns, and often include darkly stained nuclii in various stages of mitosis. The capillary in the figure is lined b~- an endothelial barrier 30, and just below that, a basement membrane 32.
A
2~67 1 78 It can be seen in Figure 13B that liposomes, such as liposomes 34, are heavily c~ncPntrated in the tumor re-gion, ad~acent the capillary on the tumor side of the endothelial barrier and basement membrane, and many lipo-5 somes are also dispersed throughout the intercellularfluid surrounding the tumor cells.
Figure 13C shows another region of the liver tumor from the above animal. Liposomes are seen throughout the intercellular fluid bathing the carcinoma cells.
In a third study, C26 colon carcinoma cells were injected s~hc~tAn~ously into an animal, and allowed to grow in the animal for 28 days. Thereafter, the animal was in~ected IV with 0.5 mg of the above liposomes.
Twenty four hours later, the animal was sacrificed, and 15 the tumor mass was exc.~sed. After --;nn, tumor mass was secti~ne~ on a microtome and stained as above.
Figure 13D shows a region of the tumor cells, including a cell 36 in the center of the figure which is ln late stage mitosis. Small, darkly stained liposomes are seen 20 throughout the intercP~ l Ar fluid.
Example 15 Tumor Treatment Method Vesicle-forming lipids cnntAin;n~ PEG-PE, PG, PHEPC, 25 and cholesterol and ~-TC in a mole ratio of 0 . 3: 0 . 3 1.4: 1: 0.2 were dissolved in chloroform to a final lipid crnrPntration of 25 ~mol rhosrhol;pirl~ml. The lipid mix-ture was dried into a thin film under reduced pressure.
The film was hydrated with a sol~t; on of .125M amm~nium 30 sulfate to form MLVs. The MLV suspension was frozen in a dry ice acetone bath and thawed three times and size~ to 80-100 nm. An Amm~n;~-m ion gradient was created substan-tially as desc_ibed in Example 10. The liposomes were loaded with epirubicin, and free ~unbound drug~ removed A
.
-6a also as ~escribed in Example 10 for doxorubicin. Thè
final concentration of entrapped drug was about 50-100 llg drug/~mol lipid. Epirubicin HCl and doxorubicin HCL, the commercial products, were obtained from the hospital 5 pharmacy.
About 10' cells C-26 colon carcinoma cells were iniected subcutaneously into three groups of 35 mice.
The groups were subdivided into 5 7-animal subgroups.
For the tumor suppression experiment shown in Figure 10 14A each subgro~p was injected IV with 0.5 ml of eithe_ saline vehicle control ~open circles), 6 mgtkg epirubicin (open triangles), 6 mg/kg doxorubicin (filled circles), or the drug-loaded liposomes (PEG-DOX liposomes~ at two doses, 6mg/kg (filled triangles) and 12 mg/kg (open 15 s~uares) on days 1, 8 and 15 following tumor cell implan-tation. Each group was followed for 28 days. Tumor size was measured for each animal on days 5,7,12,14,17,21,24 and 28. The growth of the tumor in each ~ubyLuu~ (ex-pressed as the mean tumor size of the individual animals) 2 0 at each time point is plotted in Figure 14A .
With reference to this figure, neither ~ree doxoru-bicin nor free epirubicin at 6 mg/kg si~n;firAntly sup-pressed tumor growth compared with the sali~e control.
In contrast, PEG liposome entrapped epirubicin hoth doses 25 si~n; f; ~Ant ly suppresses tumor growth. With -espect to survival of the animals at 120 days followlng tumor lmplAntat; c-n, none of the animals in the saline, epiru-bicin or doxorubicin groups survived whereas 5 out of the seven and seven out of seven survived in the 6 ~g/kg 30 liposome epirubicin and 12 mg/kg 1 ipoc~ - epirubicin groups, respectively.
- ~ The results of delayed treatment experiments using the same tumor model are presented in Figure 14B and 14C.
The same number of animals were inoculated with the same _ _ _ _ _ _ _ _ . _ .. _ _ . .... . ..
number of tumor cells as described above. The treatme~,~
groups in Figures 14B and 14C consisted of sa~_ne (solid line), 6 mg/kg epiru~icin (filled triangles), 6 mg/kg free epirubicin plus empty PEG liposomes (open circles) 5 and two doses of epirubicin entrapped in PEG liposomes, 6 mg~kg (filled triangles) and 9 mg/kg (open squares~. In contrast to the results presented in Figure 14A, only two treatments were given in these experiments: days 3 and 10 for the results plotted in Figure 14B; and days 10 and 10 17 for the results plotted in FLgure 14C. Importantly, in the case of the PEG liposome entrapped drug, both delayed treatment schedules at both dose levels result in tumor regreSSiOn whereas the free drug and free drug plus empty liposome treatment ~roups show only a mo~est retar-~5 dation in the rate of t~mor growth.
Example 16 Tumor Treatment Method PEG-DOX liposomes were prepared as in Example 15 20 except that doxorubicin was loaded in the liposomes to a final level of 60-80 ug/umoles total lipid. ~ doxorubi-cin HCl solution to be used as the free drug control was obtained from a hospital pharmacy. A total of 30 mice were in~ected IP with l0' J-6456 lymphoma cells. The 25 animals were divided into three l0-animal group~;, each of which was in~ected IV with 0 . 4 ml of either saline vehi-cle, 10 mg/kg doxorubicin solution or the doxorubicin-loaded liposomes at l0 mg/kg. Each group was rollowed ~or l00 days for number of surviving animals. The per-30 cent survivors for each treatment group is plotted inFigure 15.
As can be seen, free drug ~filled circles) provided little improvement in survival over the saline group (filled squares). In the animals treated with doxorubi-WO 91/05546 PCr/VS90/06211 ~ ,6~
~ 70 cin loaded PÉG-liposomes (filled triangles), however, about 50% of the animals survived over 40 days, 20% over 70 days, and 1096 survived until the experiment was ter-minated at 10 0 days .
Example 17 Reduced Toxicity of PEG-Liposomes Solutions of free doxorubicin HCl, epirubicin HCl were obtained as above. PEG-liposome formulations con-10 taining either doxorubicin or epirubicin, at a drugconcentration of 70-90 ug compound/umole liposome lipid, were prepared as described in EXample 16. Conventional liposomes (no PEG-derivatized lipid) were loaded with doxorubicin to a drug concentration of 40 ug/umole lipid 15 using standard t~ hn~ 5.
Each of the five f~ t t ons was administered to 35 mice, at a dose between 10 and 40 mg drug/kg body weight, in 5 mg/kG in~ - c, with five receiving each dosage.
The maximum tolerated dose given in Table 11 below is 20 highest dose which did not cause death or dramatic weight loss in the injected animals within 14 days. As seen from the data, both DOX-liposomes and PEG-DOX liposomes more than doubled the tolerated dose of doxorubicin over the drug in free form, with the PEG-DOX liposomes giving 25 a slightly higher tolerated dose. A similar result was obtained for doses of tolerated epirubicin in free and -lipl~so~al ~
Table 1 1 Maximum Tolerated Dose of DXN
(mg~Kg in mice) S DX~ 10-12 DoX-Lip 25-30 PEG-DXN-Lip 25-35 ~;P I 10 P~G-EPI 20 Example 18 Tumor Treatment ~qethod Conventional doxorubicin liposomes (L-DOX) were pre-pared according to publlshed methods. Briefly, a mixture of eggPG, Egg, PC, cholesterol and a-TC in a mole ratio of 0.3: 1.4: 1: 0.2 was made in chlorsform. The solvent was removed under reduced presssure and the dry lipld film hydrated with a solution of 155 mN NaCl rnnt~;n~n~ 2-5 mg doxorubicin HCl. The resulting ~5LV preparation was down-sized by extrusion through a series of polycarbonate membranes to a final size of about 250 nm. The free (~nentrapped) drug was remoYed by passing the suspension over a bed of Dowex resin. The final doxorubicin con-centration was about 40 per umole lipid.
Three groups of 7 mice were inoculated subcutane~us-ly with 10' - 10' C-26 colon carcinoma cells as detailed in Example 15. The animals were divided into ~hree, 7-ani~al treatment groups, one of which receivd 0.5 ml of saline vehicle as a control. The other two groups were treated with doxorubicin either as a free drug solution or in the form of L-DOX liposom~es at a dose of 10 mg,'kg.
The tret .~ s were given on days 8, 15 and 22 after tumor cell inoc~ t; nn . Tumor size was measured on the days tre~rmPnts were given and day 2B. As shown in Figure 16, the free druy (filled circles) suppressed tumor growth to a modest extent compared with the saline control (;o~id line). The tumor in the L-Dox-treated group (filled triangles ) grew slightly faster than the ~ree-dru~-treated group and slightly more slowly than in the untreate~ group . These results l nrl~ ~Ate that the 5 anti-tumor activity o~ the L-DOX preparation is about the same, and certainly no better than the same- dose of free drug. This stands in marked contrast to the results presented in Example 15 (and Figures 14A-C) which ~how that at comparable doses epirubicin entrapped in PEG-lO liposomes has dramatically better anti-tumor activity than ree dn~g 1D tbis Jame tumo:: model.
.
!
'~ .
A~
Claims (8)
PROPERTY OR PRIVILEGE IS CLAIMED ARE DEFINED AS FOLLOWS:
1. A liposome composition for use in localizing a tumor-imaging agent or an anti-tumor agent in a solid tumor via the bloodstream, the composition comprising, liposomes composed of vesicle-forming lipids and be-tween 1-20 mole percent of an amphipathic vesicle-form-ing lipid derivatized with a hydrophilic polymer selected from polyethyleneglycol, polylactic acid, polyglycolic acid and polylactic acid/polyglycolic acid copolymers having a molecular weight between 1,000-5,000 daltons and having a mean liposome size of between about 0.07-0.12 µm, and a tumor-imaging agent or an anti-tumor agent in liposome-entrapped form.
2. The liposome composition according to claim 1, wherein the hydrophilic polymer is polyethyleneglyol having a molecular weight of about 1,000 to 5,000 dal-tons.
3. The liposome composition according to claim 1 or claim 2,. wherein at least about 80% of the anti-tumor agent is in liposome-entrapped form.
4. The liposome composition according to claim 3, wherein the anti-tumor agent is an anthracycline antibi-otic.
5. The liposome composition according to claim 4, wherein the anthracycline is doxorubicin, epirubicin, and daunorubicin and pharmacologically acceptable salts and acids thereof.
6. The liposome composition according to claim 4 or 5, wherein the concentration of anti-tumor agent which is entrapped in the liposomes is greater than 50µg agent/µmole liposome lipid.
7. The use of a liposome composition comprising liposomes composed of vesicle-forming lipids and between 1-20 mole percent of an amphipathic vesicle-forming lipid derivatized with a hydrophilic polymer selected and hav-ing a mean liposome size of between about 0.07-0.12 µm, and a tumor-imaging agent or an anti-tumor agent in lipo-some-entrapped form, in the manufacture of a medicament for the localization of said agent in a solid tumor.
8. A method of preparing a tumor-imaging agent or an anti-tumor agent for localization in a solid tumor via the bloodstream, comprising entrapping the tumor-imaging agent or anti-tumor agent in liposomes to form a liposom-al composition according to any one of claims 1 to 6.
Applications Claiming Priority (2)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
US07/425,224 US5013556A (en) | 1989-10-20 | 1989-10-20 | Liposomes with enhanced circulation time |
US425,224 | 1989-10-20 |
Publications (2)
Publication Number | Publication Date |
---|---|
CA2067178A1 CA2067178A1 (en) | 1991-04-21 |
CA2067178C true CA2067178C (en) | 1997-03-25 |
Family
ID=23685679
Family Applications (2)
Application Number | Title | Priority Date | Filing Date |
---|---|---|---|
CA002067178A Expired - Lifetime CA2067178C (en) | 1989-10-20 | 1990-10-19 | Solid tumor treatment method and composition |
CA002067133A Expired - Lifetime CA2067133C (en) | 1989-10-20 | 1990-10-19 | Liposome microreservoir composition and method |
Family Applications After (1)
Application Number | Title | Priority Date | Filing Date |
---|---|---|---|
CA002067133A Expired - Lifetime CA2067133C (en) | 1989-10-20 | 1990-10-19 | Liposome microreservoir composition and method |
Country Status (18)
Country | Link |
---|---|
US (2) | US5013556A (en) |
EP (2) | EP0496813B1 (en) |
JP (4) | JP2667051B2 (en) |
KR (2) | KR920703013A (en) |
AT (2) | ATE115401T1 (en) |
AU (2) | AU642679B2 (en) |
CA (2) | CA2067178C (en) |
DE (3) | DE19675048I2 (en) |
DK (1) | DK0496835T3 (en) |
ES (1) | ES2071976T3 (en) |
FI (2) | FI105151B (en) |
GR (1) | GR3017060T3 (en) |
HK (1) | HK14097A (en) |
IL (2) | IL96069A (en) |
LU (1) | LU88854I2 (en) |
NL (1) | NL960031I2 (en) |
NO (3) | NO304637B1 (en) |
WO (2) | WO1991005546A1 (en) |
Families Citing this family (1596)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
US5456663A (en) * | 1984-05-25 | 1995-10-10 | Lemelson; Jerome H. | Drugs and methods for treating diseases |
US5882330A (en) * | 1984-05-25 | 1999-03-16 | Lemelson; Jerome H. | Drugs and methods for treating diseases |
US5925375A (en) * | 1987-05-22 | 1999-07-20 | The Liposome Company, Inc. | Therapeutic use of multilamellar liposomal prostaglandin formulations |
JPH0720857B2 (en) * | 1988-08-11 | 1995-03-08 | テルモ株式会社 | Liposome and its manufacturing method |
GB8824593D0 (en) * | 1988-10-20 | 1988-11-23 | Royal Free Hosp School Med | Liposomes |
US6132763A (en) * | 1988-10-20 | 2000-10-17 | Polymasc Pharmaceuticals Plc | Liposomes |
US5153000A (en) * | 1988-11-22 | 1992-10-06 | Kao Corporation | Phosphate, liposome comprising the phosphate as membrane constituent, and cosmetic and liposome preparation comprising the liposome |
US5620689A (en) * | 1989-10-20 | 1997-04-15 | Sequus Pharmaceuuticals, Inc. | Liposomes for treatment of B-cell and T-cell disorders |
US5527528A (en) * | 1989-10-20 | 1996-06-18 | Sequus Pharmaceuticals, Inc. | Solid-tumor treatment method |
US5013556A (en) * | 1989-10-20 | 1991-05-07 | Liposome Technology, Inc. | Liposomes with enhanced circulation time |
US5843473A (en) * | 1989-10-20 | 1998-12-01 | Sequus Pharmaceuticals, Inc. | Method of treatment of infected tissues |
US5356633A (en) * | 1989-10-20 | 1994-10-18 | Liposome Technology, Inc. | Method of treatment of inflamed tissues |
US20010051183A1 (en) * | 1989-10-20 | 2001-12-13 | Alza Corporation | Liposomes with enhanced circulation time and method of treatment |
US5469854A (en) * | 1989-12-22 | 1995-11-28 | Imarx Pharmaceutical Corp. | Methods of preparing gas-filled liposomes |
US6001335A (en) * | 1989-12-22 | 1999-12-14 | Imarx Pharmaceutical Corp. | Contrasting agents for ultrasonic imaging and methods for preparing the same |
US6088613A (en) * | 1989-12-22 | 2000-07-11 | Imarx Pharmaceutical Corp. | Method of magnetic resonance focused surgical and therapeutic ultrasound |
US5542935A (en) * | 1989-12-22 | 1996-08-06 | Imarx Pharmaceutical Corp. | Therapeutic delivery systems related applications |
US5585112A (en) * | 1989-12-22 | 1996-12-17 | Imarx Pharmaceutical Corp. | Method of preparing gas and gaseous precursor-filled microspheres |
US5580575A (en) * | 1989-12-22 | 1996-12-03 | Imarx Pharmaceutical Corp. | Therapeutic drug delivery systems |
US6146657A (en) | 1989-12-22 | 2000-11-14 | Imarx Pharmaceutical Corp. | Gas-filled lipid spheres for use in diagnostic and therapeutic applications |
US5705187A (en) * | 1989-12-22 | 1998-01-06 | Imarx Pharmaceutical Corp. | Compositions of lipids and stabilizing materials |
US5773024A (en) * | 1989-12-22 | 1998-06-30 | Imarx Pharmaceutical Corp. | Container with multi-phase composition for use in diagnostic and therapeutic applications |
US5922304A (en) | 1989-12-22 | 1999-07-13 | Imarx Pharmaceutical Corp. | Gaseous precursor filled microspheres as magnetic resonance imaging contrast agents |
US5352435A (en) * | 1989-12-22 | 1994-10-04 | Unger Evan C | Ionophore containing liposomes for ultrasound imaging |
US5305757A (en) * | 1989-12-22 | 1994-04-26 | Unger Evan C | Gas filled liposomes and their use as ultrasonic contrast agents |
US6551576B1 (en) | 1989-12-22 | 2003-04-22 | Bristol-Myers Squibb Medical Imaging, Inc. | Container with multi-phase composition for use in diagnostic and therapeutic applications |
US5776429A (en) * | 1989-12-22 | 1998-07-07 | Imarx Pharmaceutical Corp. | Method of preparing gas-filled microspheres using a lyophilized lipids |
US5441746A (en) * | 1989-12-22 | 1995-08-15 | Molecular Bioquest, Inc. | Electromagnetic wave absorbing, surface modified magnetic particles for use in medical applications, and their method of production |
US5656211A (en) * | 1989-12-22 | 1997-08-12 | Imarx Pharmaceutical Corp. | Apparatus and method for making gas-filled vesicles of optimal size |
US5733572A (en) * | 1989-12-22 | 1998-03-31 | Imarx Pharmaceutical Corp. | Gas and gaseous precursor filled microspheres as topical and subcutaneous delivery vehicles |
US5882678A (en) * | 1990-01-12 | 1999-03-16 | The Liposome Co, Inc. | Interdigitation-fusion liposomes containing arachidonic acid metabolites |
US5162361A (en) * | 1990-04-10 | 1992-11-10 | The United States Of America As Represented By The Secretary, Department Of Health And Human Services | Method of treating diseases associated with elevated levels of interleukin 1 |
US20060084797A1 (en) * | 1990-06-11 | 2006-04-20 | Gilead Sciences, Inc. | High affinity TGFbeta nucleic acid ligands and inhibitors |
US6465188B1 (en) * | 1990-06-11 | 2002-10-15 | Gilead Sciences, Inc. | Nucleic acid ligand complexes |
US20030113369A1 (en) * | 1991-01-16 | 2003-06-19 | Martin Francis J. | Liposomes with enhanced circulation time and method of treatment |
US5205290A (en) | 1991-04-05 | 1993-04-27 | Unger Evan C | Low density microspheres and their use as contrast agents for computed tomography |
US5874062A (en) * | 1991-04-05 | 1999-02-23 | Imarx Pharmaceutical Corp. | Methods of computed tomography using perfluorocarbon gaseous filled microspheres as contrast agents |
JP3220180B2 (en) * | 1991-05-23 | 2001-10-22 | 三菱化学株式会社 | Drug-containing protein-bound liposomes |
GB9111611D0 (en) * | 1991-05-30 | 1991-07-24 | Sandoz Ltd | Liposomes |
EP1236473A3 (en) | 1992-04-03 | 2003-01-15 | The Regents Of The University Of California | Self-assembling polynucleotide delivery system |
ZA933926B (en) * | 1992-06-17 | 1994-01-03 | Amgen Inc | Polyoxymethylene-oxyethylene copolymers in conjuction with blomolecules |
WO1994003501A1 (en) * | 1992-08-05 | 1994-02-17 | Meito Sangyo Kabushiki Kaisha | Small-diameter composite composed of water-soluble carboxypolysaccharide and magnetic iron oxide |
NL9201722A (en) * | 1992-10-05 | 1994-05-02 | Stichting Tech Wetenschapp | Pharmaceutical composition for the site-bound release of a clot dissolving protein and method for its preparation. |
DE4236237A1 (en) * | 1992-10-27 | 1994-04-28 | Behringwerke Ag | Prodrugs, their preparation and use as medicines |
US6764693B1 (en) * | 1992-12-11 | 2004-07-20 | Amaox, Ltd. | Free radical quenching composition and a method to increase intracellular and/or extracellular antioxidants |
JP3351476B2 (en) * | 1993-01-22 | 2002-11-25 | 三菱化学株式会社 | Phospholipid derivatives and liposomes containing the same |
US5395619A (en) * | 1993-03-03 | 1995-03-07 | Liposome Technology, Inc. | Lipid-polymer conjugates and liposomes |
US6326353B1 (en) * | 1993-03-23 | 2001-12-04 | Sequus Pharmaceuticals, Inc. | Enhanced circulation effector composition and method |
WO1994021281A1 (en) * | 1993-03-23 | 1994-09-29 | Liposome Technology, Inc. | Polymer-polypeptide composition and method |
US6180134B1 (en) * | 1993-03-23 | 2001-01-30 | Sequus Pharmaceuticals, Inc. | Enhanced ciruclation effector composition and method |
AU6785094A (en) * | 1993-05-07 | 1994-12-12 | Sequus Pharmaceuticals, Inc. | Subcutaneous liposome delivery method |
US5681811A (en) * | 1993-05-10 | 1997-10-28 | Protein Delivery, Inc. | Conjugation-stabilized therapeutic agent compositions, delivery and diagnostic formulations comprising same, and method of making and using the same |
US20050181976A1 (en) * | 1993-05-10 | 2005-08-18 | Ekwuribe Nnochiri N. | Amphiphilic oligomers |
US5359030A (en) * | 1993-05-10 | 1994-10-25 | Protein Delivery, Inc. | Conjugation-stabilized polypeptide compositions, therapeutic delivery and diagnostic formulations comprising same, and method of making and using the same |
US6191105B1 (en) | 1993-05-10 | 2001-02-20 | Protein Delivery, Inc. | Hydrophilic and lipophilic balanced microemulsion formulations of free-form and/or conjugation-stabilized therapeutic agents such as insulin |
EP1118326A3 (en) * | 1993-05-21 | 2002-07-31 | The Liposome Company, Inc. | Reduction of liposome-induced adverse physiological reactions |
US5744155A (en) * | 1993-08-13 | 1998-04-28 | Friedman; Doron | Bioadhesive emulsion preparations for enhanced drug delivery |
US5514670A (en) * | 1993-08-13 | 1996-05-07 | Pharmos Corporation | Submicron emulsions for delivery of peptides |
ES2146668T3 (en) * | 1993-10-25 | 2000-08-16 | Liposome Co Inc | LIPOSOMIC DEFENSINS. |
US5415869A (en) * | 1993-11-12 | 1995-05-16 | The Research Foundation Of State University Of New York | Taxol formulation |
US5468499A (en) * | 1993-11-15 | 1995-11-21 | Ohio State University | Liposomes containing the salt of phosphoramide mustard and related compounds |
US7083572B2 (en) * | 1993-11-30 | 2006-08-01 | Bristol-Myers Squibb Medical Imaging, Inc. | Therapeutic delivery systems |
JPH09506866A (en) * | 1993-12-14 | 1997-07-08 | ジョーンズ ホプキンス ユニバーシティー スクール オブ メディシン | Controlled release of pharmaceutically active substances for immunotherapy |
US6773719B2 (en) * | 1994-03-04 | 2004-08-10 | Esperion Luv Development, Inc. | Liposomal compositions, and methods of using liposomal compositions to treat dislipidemias |
US6312719B1 (en) * | 1994-03-04 | 2001-11-06 | The University Of British Columbia | Liposome compositions and methods for the treatment of atherosclerosis |
US20010055581A1 (en) * | 1994-03-18 | 2001-12-27 | Lawrence Tamarkin | Composition and method for delivery of biologically-active factors |
US5736121A (en) * | 1994-05-23 | 1998-04-07 | Imarx Pharmaceutical Corp. | Stabilized homogenous suspensions as computed tomography contrast agents |
DE4423131A1 (en) * | 1994-07-01 | 1996-01-04 | Bayer Ag | New hIL-4 mutant proteins as antagonists or partial agonists of human interleukin 4 |
US5626862A (en) | 1994-08-02 | 1997-05-06 | Massachusetts Institute Of Technology | Controlled local delivery of chemotherapeutic agents for treating solid tumors |
US6132764A (en) | 1994-08-05 | 2000-10-17 | Targesome, Inc. | Targeted polymerized liposome diagnostic and treatment agents |
US5512294A (en) * | 1994-08-05 | 1996-04-30 | Li; King C. | Targeted polymerized liposome contrast agents |
US6001968A (en) | 1994-08-17 | 1999-12-14 | The Rockefeller University | OB polypeptides, modified forms and compositions |
US6124448A (en) * | 1994-08-17 | 2000-09-26 | The Rockfeller University | Nucleic acid primers and probes for the mammalian OB gene |
US6124439A (en) * | 1994-08-17 | 2000-09-26 | The Rockefeller University | OB polypeptide antibodies and method of making |
US6350730B1 (en) | 1994-08-17 | 2002-02-26 | The Rockefeller University | OB polypeptides and modified forms as modulators of body weight |
US6471956B1 (en) | 1994-08-17 | 2002-10-29 | The Rockefeller University | Ob polypeptides, modified forms and compositions thereto |
US6309853B1 (en) | 1994-08-17 | 2001-10-30 | The Rockfeller University | Modulators of body weight, corresponding nucleic acids and proteins, and diagnostic and therapeutic uses thereof |
US6048837A (en) * | 1994-08-17 | 2000-04-11 | The Rockefeller University | OB polypeptides as modulators of body weight |
US5885613A (en) * | 1994-09-30 | 1999-03-23 | The University Of British Columbia | Bilayer stabilizing components and their use in forming programmable fusogenic liposomes |
US5820873A (en) * | 1994-09-30 | 1998-10-13 | The University Of British Columbia | Polyethylene glycol modified ceramide lipids and liposome uses thereof |
US5785992A (en) * | 1994-09-30 | 1998-07-28 | Inex Pharmaceuticals Corp. | Compositions for the introduction of polyanionic materials into cells |
JPH10507450A (en) * | 1994-10-14 | 1998-07-21 | ザ リポソーム カンパニー、インコーポレーテッド | Ether lipid liposomes and their therapeutic use |
CN1080575C (en) * | 1994-10-14 | 2002-03-13 | 蛋白质传送股份有限公司 | Conjugation-stabilized polypeptide compositions, therapeutic delivery and diagnostic formulations comprising same, and method of making and using the same |
US6214388B1 (en) * | 1994-11-09 | 2001-04-10 | The Regents Of The University Of California | Immunoliposomes that optimize internalization into target cells |
US6743779B1 (en) | 1994-11-29 | 2004-06-01 | Imarx Pharmaceutical Corp. | Methods for delivering compounds into a cell |
US5827531A (en) * | 1994-12-02 | 1998-10-27 | The United States Of America As Represented By The Administrator Of The National Aeronautics And Space Administration | Microcapsules and methods for making |
US5795587A (en) * | 1995-01-23 | 1998-08-18 | University Of Pittsburgh | Stable lipid-comprising drug delivery complexes and methods for their production |
US6008202A (en) * | 1995-01-23 | 1999-12-28 | University Of Pittsburgh | Stable lipid-comprising drug delivery complexes and methods for their production |
US5830430A (en) * | 1995-02-21 | 1998-11-03 | Imarx Pharmaceutical Corp. | Cationic lipids and the use thereof |
IL112834A (en) * | 1995-03-01 | 2000-12-06 | Yeda Res & Dev | Pharmaceutical compositions for controlled release of soluble receptors |
US5658588A (en) * | 1995-03-31 | 1997-08-19 | University Of Cincinnati | Fibrinogen-coated liposomes |
GB9509016D0 (en) * | 1995-05-03 | 1995-06-21 | Royal Free Hosp School Med | Tissue entrapment |
US8071737B2 (en) * | 1995-05-04 | 2011-12-06 | Glead Sciences, Inc. | Nucleic acid ligand complexes |
US5997898A (en) * | 1995-06-06 | 1999-12-07 | Imarx Pharmaceutical Corp. | Stabilized compositions of fluorinated amphiphiles for methods of therapeutic delivery |
US6420549B1 (en) | 1995-06-06 | 2002-07-16 | Isis Pharmaceuticals, Inc. | Oligonucleotide analogs having modified dimers |
US6673364B1 (en) | 1995-06-07 | 2004-01-06 | The University Of British Columbia | Liposome having an exchangeable component |
US6521211B1 (en) | 1995-06-07 | 2003-02-18 | Bristol-Myers Squibb Medical Imaging, Inc. | Methods of imaging and treatment with targeted compositions |
EP0832271B8 (en) * | 1995-06-07 | 2005-03-02 | INEX Pharmaceuticals Corp. | Lipid-nucleic acid particles prepared via a hydrophobic lipid-nucleic acid complex intermediate and use for gene transfer |
US6139819A (en) | 1995-06-07 | 2000-10-31 | Imarx Pharmaceutical Corp. | Targeted contrast agents for diagnostic and therapeutic use |
US6231834B1 (en) | 1995-06-07 | 2001-05-15 | Imarx Pharmaceutical Corp. | Methods for ultrasound imaging involving the use of a contrast agent and multiple images and processing of same |
US6033645A (en) * | 1996-06-19 | 2000-03-07 | Unger; Evan C. | Methods for diagnostic imaging by regulating the administration rate of a contrast agent |
US5981501A (en) * | 1995-06-07 | 1999-11-09 | Inex Pharmaceuticals Corp. | Methods for encapsulating plasmids in lipid bilayers |
US7422902B1 (en) | 1995-06-07 | 2008-09-09 | The University Of British Columbia | Lipid-nucleic acid particles prepared via a hydrophobic lipid-nucleic acid complex intermediate and use for gene transfer |
JPH11511128A (en) * | 1995-08-01 | 1999-09-28 | ノバルティス・アクチエンゲゼルシャフト | Liposome oligonucleotide composition |
US6107332A (en) | 1995-09-12 | 2000-08-22 | The Liposome Company, Inc. | Hydrolysis-promoting hydrophobic taxane derivatives |
US6051600A (en) * | 1995-09-12 | 2000-04-18 | Mayhew; Eric | Liposomal hydrolysis-promoting hydrophobic taxane derivatives |
US5834025A (en) | 1995-09-29 | 1998-11-10 | Nanosystems L.L.C. | Reduction of intravenously administered nanoparticulate-formulation-induced adverse physiological reactions |
CA2231547A1 (en) * | 1995-10-11 | 1997-04-17 | Kevin Jon Williams | Liposomal compositions and methods of using them |
US5858397A (en) * | 1995-10-11 | 1999-01-12 | University Of British Columbia | Liposomal formulations of mitoxantrone |
US20030040467A1 (en) * | 1998-06-15 | 2003-02-27 | Mary Ann Pelleymounter | Ig/ob fusions and uses thereof. |
US6936439B2 (en) * | 1995-11-22 | 2005-08-30 | Amgen Inc. | OB fusion protein compositions and methods |
US5626654A (en) | 1995-12-05 | 1997-05-06 | Xerox Corporation | Ink compositions containing liposomes |
US5817856A (en) * | 1995-12-11 | 1998-10-06 | Yissum Research Development Company Of The Hebrew University Of Jerusalem | Radiation-protective phospholipid and method |
US20050025742A1 (en) * | 1996-01-08 | 2005-02-03 | Canji, Inc. | Methods and compositions for interferon therapy |
US20040014709A1 (en) * | 1996-01-08 | 2004-01-22 | Canji, Inc. | Methods and compositions for interferon therapy |
US6392069B2 (en) | 1996-01-08 | 2002-05-21 | Canji, Inc. | Compositions for enhancing delivery of nucleic acids to cells |
US7002027B1 (en) | 1996-01-08 | 2006-02-21 | Canji, Inc. | Compositions and methods for therapeutic use |
US5789244A (en) * | 1996-01-08 | 1998-08-04 | Canji, Inc. | Compositions and methods for the treatment of cancer using recombinant viral vector delivery systems |
WO1997025425A1 (en) | 1996-01-08 | 1997-07-17 | Genentech, Inc. | Wsx receptor and ligands |
US6096336A (en) * | 1996-01-30 | 2000-08-01 | The Stehlin Foundation For Cancer Research | Liposomal prodrugs comprising derivatives of camptothecin and methods of treating cancer using these prodrugs |
US5908624A (en) * | 1996-06-27 | 1999-06-01 | Albany Medical College | Antigenic modulation of cells |
US5942246A (en) * | 1996-02-16 | 1999-08-24 | The Liposome Company, Inc. | Etherlipid containing multiple lipid liposomes |
US5965159A (en) * | 1996-02-16 | 1999-10-12 | The Liposome Company, Inc. | Etherlipid-containing multiple lipid liposomes |
USRE39042E1 (en) * | 1996-02-16 | 2006-03-28 | The Liposome Company, Inc. | Etherlipid-containing multiple lipid liposomes |
US6007839A (en) * | 1996-02-16 | 1999-12-28 | The Liposome Company, Inc. | Preparation of pharmaceutical compositions containing etherlipid-containing multiple lipid liposomes |
US6667053B1 (en) | 1996-02-16 | 2003-12-23 | Elan Pharmaceuticals, Inc. | D and L etherlipid stereoisomers and liposomes |
WO1997035560A1 (en) | 1996-03-28 | 1997-10-02 | The Board Of Trustees Of The University Of Illinois | Materials and methods for making improved echogenic liposome compositions |
JP2001507207A (en) | 1996-05-01 | 2001-06-05 | イマアーレクス・フアーマシユーチカル・コーポレーシヨン | Methods for delivering compounds to cells |
US20050042647A1 (en) * | 1996-06-06 | 2005-02-24 | Baker Brenda F. | Phosphorous-linked oligomeric compounds and their use in gene modulation |
US5898031A (en) | 1996-06-06 | 1999-04-27 | Isis Pharmaceuticals, Inc. | Oligoribonucleotides for cleaving RNA |
US7812149B2 (en) | 1996-06-06 | 2010-10-12 | Isis Pharmaceuticals, Inc. | 2′-Fluoro substituted oligomeric compounds and compositions for use in gene modulations |
US9096636B2 (en) | 1996-06-06 | 2015-08-04 | Isis Pharmaceuticals, Inc. | Chimeric oligomeric compounds and their use in gene modulation |
US20070275921A1 (en) * | 1996-06-06 | 2007-11-29 | Isis Pharmaceuticals, Inc. | Oligomeric Compounds That Facilitate Risc Loading |
US5919480A (en) | 1996-06-24 | 1999-07-06 | Yissum Research Development Company Of The Hebrew University Of Jerusalem | Liposomal influenza vaccine composition and method |
US8007784B1 (en) | 1996-06-27 | 2011-08-30 | Albany Medical College | Antigenic modulation of cells |
US7368129B1 (en) | 1996-08-14 | 2008-05-06 | Nutrimed Biotech | Amphiphilic materials and liposome formulations thereof |
US6284267B1 (en) | 1996-08-14 | 2001-09-04 | Nutrimed Biotech | Amphiphilic materials and liposome formulations thereof |
US5851984A (en) * | 1996-08-16 | 1998-12-22 | Genentech, Inc. | Method of enhancing proliferation or differentiation of hematopoietic stem cells using Wnt polypeptides |
US6159462A (en) * | 1996-08-16 | 2000-12-12 | Genentech, Inc. | Uses of Wnt polypeptides |
ES2208946T3 (en) * | 1996-08-23 | 2004-06-16 | Sequus Pharmaceuticals, Inc. | LIPOSOMES CONTAINING A CISPLATIN COMPOUND. |
US6271208B1 (en) | 1996-08-26 | 2001-08-07 | Transgene S.A. | Process of making cationic lipid-nucleic acid complexes |
US6414139B1 (en) | 1996-09-03 | 2002-07-02 | Imarx Therapeutics, Inc. | Silicon amphiphilic compounds and the use thereof |
CA2263568C (en) * | 1996-09-11 | 2008-12-02 | Imarx Pharmaceutical Corp. | Methods for diagnostic imaging using a contrast agent and a renal vasodilator |
US5846517A (en) * | 1996-09-11 | 1998-12-08 | Imarx Pharmaceutical Corp. | Methods for diagnostic imaging using a renal contrast agent and a vasodilator |
US6056973A (en) * | 1996-10-11 | 2000-05-02 | Sequus Pharmaceuticals, Inc. | Therapeutic liposome composition and method of preparation |
EP0932390A1 (en) | 1996-10-11 | 1999-08-04 | Sequus Pharmaceuticals, Inc. | Therapeutic liposome composition and method |
US6224903B1 (en) | 1996-10-11 | 2001-05-01 | Sequus Pharmaceuticals, Inc. | Polymer-lipid conjugate for fusion of target membranes |
TW520297B (en) * | 1996-10-11 | 2003-02-11 | Sequus Pharm Inc | Fusogenic liposome composition and method |
US6339069B1 (en) * | 1996-10-15 | 2002-01-15 | Elan Pharmaceuticalstechnologies, Inc. | Peptide-lipid conjugates, liposomes and lipsomal drug delivery |
US6087325A (en) * | 1996-10-15 | 2000-07-11 | The Liposome Company, Inc. | Peptide-lipid conjugates |
US6261537B1 (en) * | 1996-10-28 | 2001-07-17 | Nycomed Imaging As | Diagnostic/therapeutic agents having microbubbles coupled to one or more vectors |
BR9712683A (en) * | 1996-10-28 | 1999-10-19 | Nyomed Imaging A S | Diagnostic and / or therapeutically active targetable agent, combined formulation, process for preparing and using it, combined formulation, and processes for generating enhanced images of a human or non-human animal body and for in vitro targeting investigation by an agent. |
US20070036722A1 (en) * | 1996-10-28 | 2007-02-15 | Pal Rongved | Separation processes |
US6331289B1 (en) * | 1996-10-28 | 2001-12-18 | Nycomed Imaging As | Targeted diagnostic/therapeutic agents having more than one different vectors |
US5827533A (en) * | 1997-02-06 | 1998-10-27 | Duke University | Liposomes containing active agents aggregated with lipid surfactants |
WO1998035828A1 (en) * | 1997-02-14 | 1998-08-20 | The Regents Of The University Of California | Lamellar gels and methods for making and regulating |
US6090800A (en) | 1997-05-06 | 2000-07-18 | Imarx Pharmaceutical Corp. | Lipid soluble steroid prodrugs |
US6537246B1 (en) | 1997-06-18 | 2003-03-25 | Imarx Therapeutics, Inc. | Oxygen delivery agents and uses for the same |
US6143276A (en) * | 1997-03-21 | 2000-11-07 | Imarx Pharmaceutical Corp. | Methods for delivering bioactive agents to regions of elevated temperatures |
US6120751A (en) | 1997-03-21 | 2000-09-19 | Imarx Pharmaceutical Corp. | Charged lipids and uses for the same |
HUP0001256A3 (en) | 1997-04-03 | 2002-12-28 | Univ Johns Hopkins Med | Biodegradable terephthalate polyester-poly(phosphate) polymers, compositions, method for making the same and using them |
EP2301580B1 (en) | 1997-04-07 | 2012-01-18 | Genentech, Inc. | Container holding anti-VEGF antibodies |
ES2236634T3 (en) | 1997-04-07 | 2005-07-16 | Genentech, Inc. | ANTI-VEGF ANTIBODIES. |
WO1998044910A1 (en) * | 1997-04-09 | 1998-10-15 | Philipp Lang | NEW TECHNIQUE TO MONITOR DRUG DELIVERY NONINVASIVELY $i(IN VIVO) |
US20020039594A1 (en) * | 1997-05-13 | 2002-04-04 | Evan C. Unger | Solid porous matrices and methods of making and using the same |
US6416740B1 (en) | 1997-05-13 | 2002-07-09 | Bristol-Myers Squibb Medical Imaging, Inc. | Acoustically active drug delivery systems |
US6287591B1 (en) * | 1997-05-14 | 2001-09-11 | Inex Pharmaceuticals Corp. | Charged therapeutic agents encapsulated in lipid particles containing four lipid components |
DE19724796A1 (en) * | 1997-06-06 | 1998-12-10 | Max Delbrueck Centrum | Antitumor therapy agents |
US6663899B2 (en) | 1997-06-13 | 2003-12-16 | Genentech, Inc. | Controlled release microencapsulated NGF formulation |
US6113947A (en) * | 1997-06-13 | 2000-09-05 | Genentech, Inc. | Controlled release microencapsulated NGF formulation |
US6217886B1 (en) | 1997-07-14 | 2001-04-17 | The Board Of Trustees Of The University Of Illinois | Materials and methods for making improved micelle compositions |
US6548047B1 (en) | 1997-09-15 | 2003-04-15 | Bristol-Myers Squibb Medical Imaging, Inc. | Thermal preactivation of gaseous precursor filled compositions |
EP2198854B1 (en) | 1997-09-18 | 2011-11-30 | Pacira Pharmaceuticals, Inc. | Sustained-release liposomal anesthetic compositions |
US6734171B1 (en) | 1997-10-10 | 2004-05-11 | Inex Pharmaceuticals Corp. | Methods for encapsulating nucleic acids in lipid bilayers |
US6083923A (en) * | 1997-10-31 | 2000-07-04 | Isis Pharmaceuticals Inc. | Liposomal oligonucleotide compositions for modulating RAS gene expression |
US7229841B2 (en) | 2001-04-30 | 2007-06-12 | Cytimmune Sciences, Inc. | Colloidal metal compositions and methods |
JP4575592B2 (en) | 1997-11-14 | 2010-11-04 | パシラ ファーマシューティカルズ インコーポレーテッド | Production of multivesicular liposomes |
US7192589B2 (en) | 1998-09-16 | 2007-03-20 | Genentech, Inc. | Treatment of inflammatory disorders with STIgMA immunoadhesins |
JP3497133B2 (en) | 1997-11-21 | 2004-02-16 | ジェネンテック・インコーポレーテッド | A-33 related antigens and their pharmacological uses |
IL136343A0 (en) * | 1997-12-04 | 2001-05-20 | Yissum Res Dev Co | Combined chemo-immunotherapy with liposomal drugs and cytokines |
US6787132B1 (en) * | 1997-12-04 | 2004-09-07 | Yissum Research Development Company Of The Hebrew University Of Jerusalem | Combined chemo-immunotherapy with liposomal drugs and cytokines |
EP1947119A3 (en) | 1997-12-12 | 2012-12-19 | Genentech, Inc. | Treatment of cancer with anti-erb2 antibodies in combination with a chemotherapeutic agent |
US6123923A (en) | 1997-12-18 | 2000-09-26 | Imarx Pharmaceutical Corp. | Optoacoustic contrast agents and methods for their use |
US20010003580A1 (en) | 1998-01-14 | 2001-06-14 | Poh K. Hui | Preparation of a lipid blend and a phospholipid suspension containing the lipid blend |
DE69941228D1 (en) | 1998-02-04 | 2009-09-17 | Genentech Inc | Use of heregulin as epithelial cell growth factor |
AU753196B2 (en) * | 1998-02-09 | 2002-10-10 | Bracco Research S.A. | Targeted delivery of biologically active media |
US6942859B2 (en) | 1998-03-13 | 2005-09-13 | University Of Southern California | Red blood cells covalently bound with polymers |
US6312685B1 (en) | 1998-03-13 | 2001-11-06 | Timothy C. Fisher | Red blood cells covalently bound with two different polyethylene glycol derivatives |
EP1064382B1 (en) | 1998-03-17 | 2008-08-20 | Genentech, Inc. | Polypeptides homologous to vegf and bmp1 |
US6433012B1 (en) * | 1998-03-25 | 2002-08-13 | Large Scale Biology Corp. | Method for inhibiting inflammatory disease |
AU3109199A (en) * | 1998-03-25 | 1999-10-18 | Large Scale Biology Corporation | Benzoates derivatives for inhibiting angiogenesis |
US6858706B2 (en) * | 1998-04-07 | 2005-02-22 | St. Jude Children's Research Hospital | Polypeptide comprising the amino acid of an N-terminal choline binding protein a truncate, vaccine derived therefrom and uses thereof |
ATE283034T1 (en) * | 1998-04-27 | 2004-12-15 | Opperbas Holding Bv | PHARMACEUTICAL COMPOSITIONS CONTAINING FACTOR VIII AND NEUTRAL LIPOSOMES |
US6986902B1 (en) * | 1998-04-28 | 2006-01-17 | Inex Pharmaceuticals Corporation | Polyanionic polymers which enhance fusogenicity |
US6448224B1 (en) | 1998-05-06 | 2002-09-10 | St. Jude Children's Research Hospital | Antibiotics and methods of using the same |
US6331407B1 (en) | 1998-05-06 | 2001-12-18 | St. Jude Children's Research Hospital | Antibiotics and methods of using the same |
US6169078B1 (en) * | 1998-05-12 | 2001-01-02 | University Of Florida | Materials and methods for the intracellular delivery of substances |
PT1076703E (en) | 1998-05-15 | 2007-10-10 | Genentech Inc | Therapeutic uses of il-17 homologous polypeptides |
EP3112468A1 (en) | 1998-05-15 | 2017-01-04 | Genentech, Inc. | Il-17 homologous polypeptides and therapeutic uses thereof |
EP1865061A3 (en) | 1998-05-15 | 2007-12-19 | Genentech, Inc. | IL-17 homologous polypeptides and therapeutic uses thereof |
CA2328498A1 (en) | 1998-06-12 | 1999-12-16 | Genentech, Inc. | Monoclonal antibodies, cross-reactive antibodies and method for producing the same____________________________________ |
US6726925B1 (en) * | 1998-06-18 | 2004-04-27 | Duke University | Temperature-sensitive liposomal formulation |
US6200598B1 (en) * | 1998-06-18 | 2001-03-13 | Duke University | Temperature-sensitive liposomal formulation |
US20040229363A1 (en) * | 1998-06-24 | 2004-11-18 | Ed Nolan | High efficiency transfection based on low electric field strength, long pulse length |
US20030022854A1 (en) * | 1998-06-25 | 2003-01-30 | Dow Steven W. | Vaccines using nucleic acid-lipid complexes |
US6693086B1 (en) * | 1998-06-25 | 2004-02-17 | National Jewish Medical And Research Center | Systemic immune activation method using nucleic acid-lipid complexes |
US20040247662A1 (en) * | 1998-06-25 | 2004-12-09 | Dow Steven W. | Systemic immune activation method using nucleic acid-lipid complexes |
US6277413B1 (en) | 1998-07-17 | 2001-08-21 | Skyepharma, Inc. | Biodegradable compositions for the controlled release of encapsulated substances |
US20020172678A1 (en) | 2000-06-23 | 2002-11-21 | Napoleone Ferrara | EG-VEGF nucleic acids and polypeptides and methods of use |
US6703381B1 (en) | 1998-08-14 | 2004-03-09 | Nobex Corporation | Methods for delivery therapeutic compounds across the blood-brain barrier |
US6355268B1 (en) * | 1998-09-16 | 2002-03-12 | Alza Corporation | Liposome-entrapped topoisomerase inhibitors |
US6077709A (en) | 1998-09-29 | 2000-06-20 | Isis Pharmaceuticals Inc. | Antisense modulation of Survivin expression |
US6153212A (en) | 1998-10-02 | 2000-11-28 | Guilford Pharmaceuticals Inc. | Biodegradable terephthalate polyester-poly (phosphonate) compositions, articles, and methods of using the same |
US6419709B1 (en) | 1998-10-02 | 2002-07-16 | Guilford Pharmaceuticals, Inc. | Biodegradable terephthalate polyester-poly(Phosphite) compositions, articles, and methods of using the same |
EP1124961B9 (en) * | 1998-10-23 | 2010-07-21 | Kirin-Amgen Inc. | Thrombopoietic compounds |
US6660843B1 (en) * | 1998-10-23 | 2003-12-09 | Amgen Inc. | Modified peptides as therapeutic agents |
EP1950300A3 (en) | 1998-11-18 | 2011-03-23 | Genentech, Inc. | Antibody variants with higher binding affinity compared to parent antibodies |
AU1740900A (en) * | 1998-11-20 | 2000-06-13 | Genentech Inc. | Method of inhibiting angiogenesis |
US6773911B1 (en) * | 1998-11-23 | 2004-08-10 | Amgen Canada Inc. | Apoptosis-inducing factor |
EP2075335A3 (en) | 1998-12-22 | 2009-09-30 | Genentech, Inc. | Methods and compositions for inhibiting neoplastic cell growth |
IL143596A0 (en) | 1998-12-22 | 2002-04-21 | Genentech Inc | Vascular endothelial cell growth factor antagonists and uses thereof |
US6350464B1 (en) | 1999-01-11 | 2002-02-26 | Guilford Pharmaceuticals, Inc. | Methods for treating ovarian cancer, poly (phosphoester) compositions, and biodegradable articles for same |
US6818227B1 (en) * | 1999-02-08 | 2004-11-16 | Alza Corporation | Liposome composition and method for administration of a radiosensitizer |
US6537585B1 (en) | 1999-03-26 | 2003-03-25 | Guilford Pharmaceuticals, Inc. | Methods and compositions for treating solid tumors |
US6379698B1 (en) | 1999-04-06 | 2002-04-30 | Isis Pharmaceuticals, Inc. | Fusogenic lipids and vesicles |
US7098192B2 (en) | 1999-04-08 | 2006-08-29 | Isis Pharmaceuticals, Inc. | Antisense oligonucleotide modulation of STAT3 expression |
US7112337B2 (en) | 1999-04-23 | 2006-09-26 | Alza Corporation | Liposome composition for delivery of nucleic acid |
JP2002543134A (en) * | 1999-04-29 | 2002-12-17 | アルザ・コーポレーション | Liposomal compositions for improving drug retention |
CA2270600A1 (en) | 1999-05-03 | 2000-11-03 | Infectio Recherche Inc. | Method and formulations for the treatment of diseases, particularly those caused by human immunodeficiency virus and leishmania |
EP1645292A1 (en) | 1999-05-07 | 2006-04-12 | Genentech, Inc. | Treatment of autoimmune diseases with antagonists which bind to B cell surface markers |
WO2000067760A1 (en) * | 1999-05-11 | 2000-11-16 | Sankyo Company, Limited | Liposome preparation of fat-soluble antitumor drug |
US20040229779A1 (en) * | 1999-05-14 | 2004-11-18 | Ramesh Kekuda | Therapeutic polypeptides, nucleic acids encoding same, and methods of use |
US6974684B2 (en) * | 2001-08-08 | 2005-12-13 | Curagen Corporation | Therapeutic polypeptides, nucleic acids encoding same, and methods of use |
US20040023874A1 (en) * | 2002-03-15 | 2004-02-05 | Burgess Catherine E. | Therapeutic polypeptides, nucleic acids encoding same, and methods of use |
US20040067490A1 (en) * | 2001-09-07 | 2004-04-08 | Mei Zhong | Therapeutic polypeptides, nucleic acids encoding same, and methods of use |
AU5152700A (en) | 1999-06-15 | 2001-01-02 | Genentech Inc. | Secreted and transmembrane polypeptides and nucleic acids encoding the same |
US7169889B1 (en) | 1999-06-19 | 2007-01-30 | Biocon Limited | Insulin prodrugs hydrolyzable in vivo to yield peglylated insulin |
US6309633B1 (en) | 1999-06-19 | 2001-10-30 | Nobex Corporation | Amphiphilic drug-oligomer conjugates with hydroyzable lipophile components and methods for making and using the same |
CA2376849C (en) * | 1999-06-24 | 2008-10-14 | Kyowa Hakko Kogyo Co., Ltd. | Method of inhibiting leakage of drug encapsulated in liposomes |
AU784045B2 (en) | 1999-06-25 | 2006-01-19 | Genentech Inc. | Humanized anti-ErbB2 antibodies and treatment with anti-ErbB2 antibodies |
US6352996B1 (en) | 1999-08-03 | 2002-03-05 | The Stehlin Foundation For Cancer Research | Liposomal prodrugs comprising derivatives of camptothecin and methods of treating cancer using these prodrugs |
GB9918670D0 (en) | 1999-08-06 | 1999-10-13 | Celltech Therapeutics Ltd | Biological product |
KR20110008112A (en) | 1999-08-27 | 2011-01-25 | 제넨테크, 인크. | Dosages for treatment with anti-erbb2 antibodies |
KR100510795B1 (en) * | 1999-08-31 | 2005-08-30 | Compositions and Methods for the Treatment of Tumor | |
ATE362374T1 (en) * | 1999-09-03 | 2007-06-15 | Amgen Inc | METHODS AND COMPOSITIONS FOR PREVENTING OR TREATING CANCER AND CANCER ASSOCIATED BONE LOSS |
AU7868400A (en) | 1999-10-08 | 2001-04-23 | Alza Corporation | Neutral-cationic lipid for nucleic acid and drug delivery |
IT1315253B1 (en) * | 1999-10-22 | 2003-02-03 | Novuspharma Spa | LIPOSOMIAL PREPARATION OF 6,9-BIS- (2-AMINOXYL) AMINO | BENZOG | ISOCHINOLIN-5,10-DIONE DIMALEATO |
US7067111B1 (en) | 1999-10-25 | 2006-06-27 | Board Of Regents, University Of Texas System | Ethylenedicysteine (EC)-drug conjugates, compositions and methods for tissue specific disease imaging |
US6511676B1 (en) * | 1999-11-05 | 2003-01-28 | Teni Boulikas | Therapy for human cancers using cisplatin and other drugs or genes encapsulated into liposomes |
AU784634B2 (en) | 1999-11-30 | 2006-05-18 | Mayo Foundation For Medical Education And Research | B7-H1, a novel immunoregulatory molecule |
JP2004522404A (en) | 1999-12-01 | 2004-07-29 | ジェネンテック・インコーポレーテッド | Secreted and transmembrane polypeptides and nucleic acids encoding them |
US6593308B2 (en) * | 1999-12-03 | 2003-07-15 | The Regents Of The University Of California | Targeted drug delivery with a hyaluronan ligand |
CA2393595A1 (en) * | 1999-12-10 | 2001-06-14 | Gigi Chiu | Lipid carrier compositions with protected surface reactive functions |
EP2163625B1 (en) | 1999-12-23 | 2014-05-21 | Genentech, Inc. | IL-17 and IL-17R homologous polypeptides and therapeutic uses thereof |
JP2003520210A (en) * | 2000-01-05 | 2003-07-02 | イマレックス セラピューティクス, インコーポレイテッド | Pharmaceutical formulations for delivery of drugs with low water solubility |
US20040009229A1 (en) * | 2000-01-05 | 2004-01-15 | Unger Evan Charles | Stabilized nanoparticle formulations of camptotheca derivatives |
EP1246917B1 (en) | 2000-01-13 | 2009-03-04 | Genentech, Inc. | Human stra6 polypeptides |
US6261840B1 (en) | 2000-01-18 | 2001-07-17 | Isis Pharmaceuticals, Inc. | Antisense modulation of PTP1B expression |
US20020055479A1 (en) | 2000-01-18 | 2002-05-09 | Cowsert Lex M. | Antisense modulation of PTP1B expression |
US20010033861A1 (en) * | 2000-01-28 | 2001-10-25 | Lasic Danilo D. | Liposomes containing an entrapped compound in supersaturated solution |
US20030176385A1 (en) * | 2000-02-15 | 2003-09-18 | Jingfang Ju | Antisense modulation of protein expression |
JP4922824B2 (en) * | 2000-04-03 | 2012-04-25 | 参天製薬株式会社 | Delivery substance and drug delivery system using the same |
JP2003531588A (en) | 2000-04-11 | 2003-10-28 | ジェネンテック・インコーポレーテッド | Multivalent antibodies and their uses |
EP1272160B1 (en) * | 2000-04-12 | 2007-01-17 | Liplasome Pharma A/S | Lipid-based drug delivery systems for topical application |
MXPA02010324A (en) | 2000-04-21 | 2003-04-25 | Amgen Inc | Apo ai aii peptide derivatives. |
US6667300B2 (en) | 2000-04-25 | 2003-12-23 | Icos Corporation | Inhibitors of human phosphatidylinositol 3-kinase delta |
US7030219B2 (en) | 2000-04-28 | 2006-04-18 | Johns Hopkins University | B7-DC, Dendritic cell co-stimulatory molecules |
US6677136B2 (en) | 2000-05-03 | 2004-01-13 | Amgen Inc. | Glucagon antagonists |
AU2001258106A1 (en) * | 2000-05-11 | 2001-11-20 | Celator Technologies Inc. | Lipid carrier compositions for improved drug retention |
US6680172B1 (en) | 2000-05-16 | 2004-01-20 | Regents Of The University Of Michigan | Treatments and markers for cancers of the central nervous system |
AU6531101A (en) | 2000-06-02 | 2001-12-17 | Genentech Inc | Secreted and transmembrane polypeptides and nucleic acids encoding the same |
CN1292798C (en) * | 2000-06-02 | 2007-01-03 | 德克萨斯州立大学董事会 | Ethylenedicysteine (EC)-drug conjugates |
US20030072794A1 (en) * | 2000-06-09 | 2003-04-17 | Teni Boulikas | Encapsulation of plasmid DNA (lipogenes™) and therapeutic agents with nuclear localization signal/fusogenic peptide conjugates into targeted liposome complexes |
JP2004512262A (en) | 2000-06-20 | 2004-04-22 | アイデック ファーマスーティカルズ コーポレイション | Non-radioactive anti-CD20 antibody / radiolabeled anti-CD22 antibody combination |
CA2648048A1 (en) | 2000-06-23 | 2002-01-03 | Genentech, Inc. | Compositions and methods for the diagnosis and treatment of disorders involving angiogenesis |
EP2042597B1 (en) | 2000-06-23 | 2014-05-07 | Genentech, Inc. | Compositions and methods for the diagnosis and treatment of disorders involving angiogenesis |
US20040023259A1 (en) * | 2000-07-26 | 2004-02-05 | Luca Rastelli | Therapeutic polypeptides, nucleic acids encoding same, and methods of use |
ATE412009T1 (en) | 2000-08-24 | 2008-11-15 | Genentech Inc | METHOD FOR INHIBITING IL-22 INDUCED PAP1 |
EP1944317A3 (en) | 2000-09-01 | 2008-09-17 | Genentech, Inc. | Secreted and transmembrane polypeptides and nucleic acids encoding the same |
US20030133972A1 (en) * | 2000-10-11 | 2003-07-17 | Targesome, Inc. | Targeted multivalent macromolecules |
CA2429625C (en) | 2000-10-18 | 2012-05-01 | Robert Sackstein | Hematopoietic cell e-selectin/l-selectin ligand polypeptides and methods of use thereof |
US7045283B2 (en) | 2000-10-18 | 2006-05-16 | The Regents Of The University Of California | Methods of high-throughput screening for internalizing antibodies |
WO2002036073A2 (en) * | 2000-11-02 | 2002-05-10 | Smithkline Beecham Corporation | Receptor antagonist-lipid conjugates and delivery vehicles containing same |
DE10056136A1 (en) * | 2000-11-07 | 2002-05-16 | Nemod New Modalities | Inhibiting leukocyte or tumor cell adhesion to vascular endothelial cells e.g. for combating inflammation or metastasis, using e.g. pregnancy proteins or selectin binding liposomes containing calcium-binding compound |
US8110218B2 (en) * | 2000-11-30 | 2012-02-07 | The Research Foundation Of State University Of New York | Compositions and methods for less immunogenic protein-lipid complexes |
WO2002043665A2 (en) * | 2000-11-30 | 2002-06-06 | The Research Foundation Of State University Of New York | Ahf associated dispersion system and method for preparation |
US20020127635A1 (en) * | 2000-11-30 | 2002-09-12 | Balasubramanian Sathyamangalam V. | Method of complexing a protein by the use of a dispersed system and proteins thereof |
US7625584B2 (en) * | 2000-11-30 | 2009-12-01 | The Research Foundation Of State University Of New York | Method of complexing a protein by the use of a dispersed system and proteins thereof |
WO2002059145A1 (en) * | 2000-12-18 | 2002-08-01 | Cubist Pharmaceuticals, Inc. | Methods for preparing purified lipopeptides |
US20060014674A1 (en) | 2000-12-18 | 2006-01-19 | Dennis Keith | Methods for preparing purified lipopeptides |
ES2691680T3 (en) * | 2000-12-18 | 2018-11-28 | Cubist Pharmaceuticals Llc | Daptomycin in crystalline form and its preparation |
US20040077604A1 (en) | 2001-12-19 | 2004-04-22 | Lenard Lichtenberger | Method and compositions employing formulations of lecithin oils and nsaids for protecting the gastrointestinal tract and providingenhanced therapeutic activity |
US6964849B2 (en) | 2001-01-11 | 2005-11-15 | Curagen Corporation | Proteins and nucleic acids encoding same |
US20040043928A1 (en) * | 2001-08-02 | 2004-03-04 | Ramesh Kekuda | Therapeutic polypeptides, nucleic acids encoding same, and methods of use |
US20020159996A1 (en) | 2001-01-31 | 2002-10-31 | Kandasamy Hariharan | Use of CD23 antagonists for the treatment of neoplastic disorders |
AR035779A1 (en) | 2001-02-06 | 2004-07-14 | Genetics Inst Llc | FUSION POLYPEPTIDES DERIVED FROM GLICOPROTEIN IB PLATE ALFA AND METHODS OF USE OF THE SAME |
US7060675B2 (en) | 2001-02-15 | 2006-06-13 | Nobex Corporation | Methods of treating diabetes mellitus |
US6867183B2 (en) * | 2001-02-15 | 2005-03-15 | Nobex Corporation | Pharmaceutical compositions of insulin drug-oligomer conjugates and methods of treating diseases therewith |
US7087726B2 (en) | 2001-02-22 | 2006-08-08 | Genentech, Inc. | Anti-interferon-α antibodies |
AU2002245629A1 (en) * | 2001-03-08 | 2002-09-24 | Targesome, Inc. | Stabilized therapeutic and imaging agents |
EP1385479A4 (en) * | 2001-03-26 | 2006-12-06 | Alza Corp | Liposome composition for improved intracellular delivery of a therapeutic agent |
US20040126900A1 (en) * | 2001-04-13 | 2004-07-01 | Barry Stephen E | High affinity peptide- containing nanoparticles |
US20030203865A1 (en) * | 2001-04-30 | 2003-10-30 | Pierrot Harvie | Lipid-comprising drug delivery complexes and methods for their production |
US20030077829A1 (en) * | 2001-04-30 | 2003-04-24 | Protiva Biotherapeutics Inc.. | Lipid-based formulations |
GB0111279D0 (en) * | 2001-05-10 | 2001-06-27 | Nycomed Imaging As | Radiolabelled liposomes |
AU2002342669C1 (en) | 2001-05-11 | 2010-10-07 | Amgen, Inc. | Peptides and related molecules that bind to TALL-1 |
JP2004528384A (en) * | 2001-05-15 | 2004-09-16 | トランジェーヌ、ソシエテ、アノニム | Complex for introducing target substances into cells |
WO2002094194A2 (en) * | 2001-05-22 | 2002-11-28 | Duke University | Compositions and methods for inhibiting metastasis |
WO2002094376A2 (en) * | 2001-05-22 | 2002-11-28 | Duke University | Compositions and methods for promoting or inhibiting ndpk |
CN1684708A (en) | 2001-05-30 | 2005-10-19 | 基因技术股份有限公司 | Anti-NGF antibodies for the treatment of various disorders |
JP2004532252A (en) * | 2001-05-31 | 2004-10-21 | スカイファーマ インコーポレーテッド | Encapsulation of nanosuspension in liposomes and microspheres |
US6828297B2 (en) * | 2001-06-04 | 2004-12-07 | Nobex Corporation | Mixtures of insulin drug-oligomer conjugates comprising polyalkylene glycol, uses thereof, and methods of making same |
US7713932B2 (en) * | 2001-06-04 | 2010-05-11 | Biocon Limited | Calcitonin drug-oligomer conjugates, and uses thereof |
US6858580B2 (en) * | 2001-06-04 | 2005-02-22 | Nobex Corporation | Mixtures of drug-oligomer conjugates comprising polyalkylene glycol, uses thereof, and methods of making same |
US6828305B2 (en) * | 2001-06-04 | 2004-12-07 | Nobex Corporation | Mixtures of growth hormone drug-oligomer conjugates comprising polyalkylene glycol, uses thereof, and methods of making same |
US6835802B2 (en) * | 2001-06-04 | 2004-12-28 | Nobex Corporation | Methods of synthesizing substantially monodispersed mixtures of polymers having polyethylene glycol moieties |
US6713452B2 (en) * | 2001-06-04 | 2004-03-30 | Nobex Corporation | Mixtures of calcitonin drug-oligomer conjugates comprising polyalkylene glycol, uses thereof, and methods of making same |
US20070160576A1 (en) | 2001-06-05 | 2007-07-12 | Genentech, Inc. | IL-17A/F heterologous polypeptides and therapeutic uses thereof |
EP1399215A4 (en) * | 2001-06-08 | 2004-12-22 | Utah Ventures Ii L P | Tissue-specific endothelial membrane proteins |
DK2000545T3 (en) | 2001-06-20 | 2011-11-28 | Genentech Inc | Compositions and methods for diagnosis and treatment of lung tumor |
US20050107595A1 (en) * | 2001-06-20 | 2005-05-19 | Genentech, Inc. | Compositions and methods for the diagnosis and treatment of tumor |
US7803915B2 (en) * | 2001-06-20 | 2010-09-28 | Genentech, Inc. | Antibody compositions for the diagnosis and treatment of tumor |
CA2451643C (en) | 2001-06-21 | 2012-11-13 | Isis Pharmaceuticals, Inc. | Antisense modulation of superoxide dismutase 1, soluble expression |
CN1827766B (en) | 2001-06-28 | 2010-08-25 | 徐荣祥 | In vitro cell cultivation method |
US20030087274A1 (en) * | 2001-07-05 | 2003-05-08 | Anderson David W. | Therapeutic polypeptides, nucleic acids encoding same, and methods of use |
US20040029790A1 (en) * | 2001-07-05 | 2004-02-12 | Meera Patturajan | Novel human proteins, polynucleotides encoding them and methods of using the same |
AU2002354957A1 (en) * | 2001-07-19 | 2003-03-03 | Guilford Pharmaceuticals, Inc. | Biocompatible polymer containing composition for treatment of prostate cancers |
WO2003007915A2 (en) * | 2001-07-19 | 2003-01-30 | Guilford Pharmaceuticals, Inc. | Compositions for treatment of head and neck cancers, and methods of making and using the same |
US7425545B2 (en) | 2001-07-25 | 2008-09-16 | Isis Pharmaceuticals, Inc. | Modulation of C-reactive protein expression |
US6964950B2 (en) | 2001-07-25 | 2005-11-15 | Isis Pharmaceuticals, Inc. | Antisense modulation of C-reactive protein expression |
IL160016A0 (en) * | 2001-07-29 | 2004-06-20 | Yissum Res Dev Co | Osteogenic growth oligopeptides as stimulants of hematopoiesis |
US20030096772A1 (en) | 2001-07-30 | 2003-05-22 | Crooke Rosanne M. | Antisense modulation of acyl CoA cholesterol acyltransferase-2 expression |
US7407943B2 (en) | 2001-08-01 | 2008-08-05 | Isis Pharmaceuticals, Inc. | Antisense modulation of apolipoprotein B expression |
US20040030096A1 (en) * | 2001-08-02 | 2004-02-12 | Linda Gorman | Novel human proteins, polynucleotides encoding them and methods of using the same |
US7227014B2 (en) | 2001-08-07 | 2007-06-05 | Isis Pharmaceuticals, Inc. | Antisense modulation of apolipoprotein (a) expression |
US20030059375A1 (en) * | 2001-08-20 | 2003-03-27 | Transave, Inc. | Method for treating lung cancers |
US20030211040A1 (en) | 2001-08-31 | 2003-11-13 | Paul Greengard | Phosphodiesterase activity and regulation of phosphodiesterase 1B-mediated signaling in brain |
US20040235068A1 (en) * | 2001-09-05 | 2004-11-25 | Levinson Arthur D. | Methods for the identification of polypeptide antigens associated with disorders involving aberrant cell proliferation and compositions useful for the treatment of such disorders |
US6913903B2 (en) | 2001-09-07 | 2005-07-05 | Nobex Corporation | Methods of synthesizing insulin polypeptide-oligomer conjugates, and proinsulin polypeptide-oligomer conjugates and methods of synthesizing same |
US7166571B2 (en) * | 2001-09-07 | 2007-01-23 | Biocon Limited | Insulin polypeptide-oligomer conjugates, proinsulin polypeptide-oligomer conjugates and methods of synthesizing same |
US7312192B2 (en) * | 2001-09-07 | 2007-12-25 | Biocon Limited | Insulin polypeptide-oligomer conjugates, proinsulin polypeptide-oligomer conjugates and methods of synthesizing same |
US7196059B2 (en) * | 2001-09-07 | 2007-03-27 | Biocon Limited | Pharmaceutical compositions of insulin drug-oligomer conjugates and methods of treating diseases therewith |
US7547673B2 (en) * | 2001-09-13 | 2009-06-16 | The Johns Hopkins University | Therapeutics for cancer using 3-bromopyruvate and other selective inhibitors of ATP production |
CA2460120A1 (en) | 2001-09-18 | 2003-03-27 | Genentech, Inc. | Compositions and methods for the diagnosis and treatment of tumor |
US7981863B2 (en) | 2001-09-19 | 2011-07-19 | Neuronova Ab | Treatment of Parkinson's disease with PDGF |
DE10148065A1 (en) * | 2001-09-28 | 2003-04-17 | Max Planck Gesellschaft | (Ester) -lysolecithins in liposomes |
CN1635873A (en) * | 2001-09-28 | 2005-07-06 | 埃斯佩里安医疗公司 | Methods and apparatus for extrusion of vesicles at high pressure |
ATE516364T1 (en) | 2001-10-09 | 2011-07-15 | Isis Pharmaceuticals Inc | ANTISENSE MODULATION OF EXPRESSION OF THE INSULIN-LIKE GROWTH FACTOR BINDING PROTEY S 5 |
US6750019B2 (en) | 2001-10-09 | 2004-06-15 | Isis Pharmaceuticals, Inc. | Antisense modulation of insulin-like growth factor binding protein 5 expression |
US20030199442A1 (en) * | 2001-10-09 | 2003-10-23 | Alsobrook John P. | Therapeutic polypeptides, nucleic acids encoding same, and methods of use |
US7332474B2 (en) * | 2001-10-11 | 2008-02-19 | Amgen Inc. | Peptides and related compounds having thrombopoietic activity |
AU2002368202B2 (en) | 2001-11-02 | 2008-06-05 | Insert Therapeutics, Inc | Methods and compositions for therapeutic use of RNA interference |
CA2472055A1 (en) * | 2001-11-07 | 2003-05-15 | Inex Pharmaceuticals Corporation | Improved mucosal vaccines and methods for using the same |
US20040219201A1 (en) * | 2001-12-06 | 2004-11-04 | Yechezkel Barenholz | Tempamine compositions and methods of use |
US6965025B2 (en) | 2001-12-10 | 2005-11-15 | Isis Pharmaceuticals, Inc. | Antisense modulation of connective tissue growth factor expression |
JP2005525095A (en) | 2002-01-02 | 2005-08-25 | ジェネンテック・インコーポレーテッド | Compositions and methods for tumor diagnosis and treatment |
JP4508645B2 (en) | 2002-01-04 | 2010-07-21 | ザ ロックフェラー ユニバーシティー | Compositions and methods for the prevention and treatment of amyloid-beta peptide related diseases |
US20040052928A1 (en) * | 2002-09-06 | 2004-03-18 | Ehud Gazit | Peptides and methods using same for diagnosing and treating amyloid-associated diseases |
US7781396B2 (en) * | 2002-01-31 | 2010-08-24 | Tel Aviv University Future Technology Development L.P. | Peptides directed for diagnosis and treatment of amyloid-associated disease |
AU2003215254A1 (en) * | 2002-02-13 | 2003-09-04 | Immunology Laboratories, Inc. | Compositions and methods for treatment of microbial infections |
JP2005535290A (en) | 2002-02-22 | 2005-11-24 | ジェネンテック・インコーポレーテッド | Compositions and methods for the treatment of immune related diseases |
US20100311954A1 (en) * | 2002-03-01 | 2010-12-09 | Xencor, Inc. | Optimized Proteins that Target Ep-CAM |
US20090042291A1 (en) * | 2002-03-01 | 2009-02-12 | Xencor, Inc. | Optimized Fc variants |
US20040132101A1 (en) | 2002-09-27 | 2004-07-08 | Xencor | Optimized Fc variants and methods for their generation |
US20030180712A1 (en) | 2002-03-20 | 2003-09-25 | Biostratum Ab | Inhibition of the beta3 subunit of L-type Ca2+ channels |
US20040009216A1 (en) * | 2002-04-05 | 2004-01-15 | Rodrigueza Wendi V. | Compositions and methods for dosing liposomes of certain sizes to treat or prevent disease |
AU2003230874A1 (en) | 2002-04-16 | 2003-11-03 | Genentech, Inc. | Compositions and methods for the diagnosis and treatment of tumor |
EP1517923A2 (en) * | 2002-04-22 | 2005-03-30 | Recopharma AB | Compositions and methods for inhibiting microbial adhesion |
US20040009944A1 (en) * | 2002-05-10 | 2004-01-15 | Inex Pharmaceuticals Corporation | Methylated immunostimulatory oligonucleotides and methods of using the same |
US7199107B2 (en) | 2002-05-23 | 2007-04-03 | Isis Pharmaceuticals, Inc. | Antisense modulation of kinesin-like 1 expression |
EP2305710A3 (en) | 2002-06-03 | 2013-05-29 | Genentech, Inc. | Synthetic antibody phage libraries |
US7601688B2 (en) * | 2002-06-13 | 2009-10-13 | Biocon Limited | Methods of reducing hypoglycemic episodes in the treatment of diabetes mellitus |
AU2003245160B2 (en) | 2002-06-28 | 2009-09-24 | Arbutus Biopharma Corporation | Method and apparatus for producing liposomes |
WO2004004635A2 (en) * | 2002-07-02 | 2004-01-15 | Board Of Regents, The University Of Texas System | Radiolabeled compounds and liposomes and their methods of making and using the same |
US20040247661A1 (en) * | 2002-07-03 | 2004-12-09 | Dov Michaeli | Liposomal vaccine |
US20050169979A1 (en) * | 2002-07-03 | 2005-08-04 | Dov Michaeli | Liposomal vaccine |
CN1665487A (en) * | 2002-07-03 | 2005-09-07 | 埃弗顿有限公司 | Liposomal vaccine |
CA2489588A1 (en) | 2002-07-08 | 2004-01-15 | Genentech, Inc. | Compositions and methods for the treatment of immune related diseases |
CN101711866A (en) | 2002-07-15 | 2010-05-26 | 健泰科生物技术公司 | Method for identifying tumors that are responsive to treatment with anti-ErbB2 antibodies |
US7622118B2 (en) * | 2002-07-15 | 2009-11-24 | Board Of Regents, The University Of Texas System | Cancer treatment methods using selected antibodies to aminophospholipids |
US7678386B2 (en) * | 2002-07-15 | 2010-03-16 | Board Of Regents The University Of Texas | Liposomes coated with selected antibodies that bind to aminophospholipids |
US7714109B2 (en) * | 2002-07-15 | 2010-05-11 | Board Of Regents, The University Of Texas System | Combinations and kits for cancer treatment using selected antibodies to aminophospholipids |
US7572448B2 (en) * | 2002-07-15 | 2009-08-11 | Board Of Regents, The University Of Texas System | Combined cancer treatment methods using selected antibodies to aminophospholipids |
US7625563B2 (en) * | 2002-07-15 | 2009-12-01 | Board Of Regents, The University Of Texas System | Cancer treatment methods using selected immunoconjugates for binding to aminophospholipids |
US7615223B2 (en) * | 2002-07-15 | 2009-11-10 | Board Of Regents, The University Of Texas System | Selected immunoconjugates for binding to aminophospholipids |
EP2263697A3 (en) * | 2002-07-15 | 2011-01-19 | Board of Regents, The University of Texas System | Duramycin peptide binding to anionic phospholipids and aminophospholipids conjugates and their use in treating viral infections |
CA2492654A1 (en) * | 2002-07-16 | 2004-01-22 | University Of South Florida | Human immunosuppressive protein |
US20040161423A1 (en) * | 2002-07-18 | 2004-08-19 | Sanjeev Kumar (Mendiratta) | Polymer modified anti-angiogenic serpins |
US20040101553A1 (en) * | 2002-08-02 | 2004-05-27 | Transave, Inc. | Platinum aggregates and process for producing the same |
US9186322B2 (en) * | 2002-08-02 | 2015-11-17 | Insmed Incorporated | Platinum aggregates and process for producing the same |
ES2381617T5 (en) | 2002-08-14 | 2016-02-24 | Macrogenics, Inc. | Specific antibodies against FcgammaRIIB and its procedures for use |
EP1393720A1 (en) * | 2002-08-27 | 2004-03-03 | Universiteit Utrecht | Vesicle-encapsulated corticosteroids for treatment of cancer |
PL376536A1 (en) | 2002-08-28 | 2006-01-09 | Immunex Corporation | Compositions and methods for treating cardiovascular disease |
US20070231379A1 (en) * | 2002-08-29 | 2007-10-04 | Slater James L | Liposome-entrapped topoisomerase inhibitors |
AR036316A1 (en) * | 2002-08-29 | 2004-08-25 | Monte Verde S A | A PHARMACEUTICAL COMPOSITION OF SMALL SIZE LIPOSOMES AND PREPARATION METHOD |
US20080214437A1 (en) * | 2002-09-06 | 2008-09-04 | Mohapatra Shyam S | Methods and compositions for reducing activity of the atrial natriuretic peptide receptor and for treatment of diseases |
US7655772B2 (en) | 2002-09-06 | 2010-02-02 | University Of South Florida | Materials and methods for treatment of allergic diseases |
CA2497334A1 (en) | 2002-09-11 | 2004-03-25 | Genentech, Inc. | Novel compositions and methods for the treatment of immune related diseases |
AU2003270439B2 (en) | 2002-09-11 | 2009-09-24 | Genentech, Inc. | Novel composition and methods for the treatment of immune related diseases |
US20050196382A1 (en) * | 2002-09-13 | 2005-09-08 | Replicor, Inc. | Antiviral oligonucleotides targeting viral families |
BR0314236A (en) * | 2002-09-13 | 2005-08-09 | Replicor Inc | Oligonucleotide formulation, pharmaceutical composition, kit, antiviral compound, preparation of oligonucleotide and methods for selection of an antiviral oligonucleotide for use as an antiviral agent, for prophylaxis or treatment of a viral infection in a patient, for prophylactic treatment of cancer caused by oncoviruses. for identifying a compound that alters the binding of an oligonucleotide to at least one viral component, for purifying oligonucleotide binding to at least one viral component and for enriching oligonucleotides from an oligonucleotide cluster |
EP2444409A2 (en) | 2002-09-16 | 2012-04-25 | Genentech, Inc. | Compositions and methods for the treatment of immune related diseases |
US6919426B2 (en) | 2002-09-19 | 2005-07-19 | Amgen Inc. | Peptides and related molecules that modulate nerve growth factor activity |
EP1585482A4 (en) | 2002-09-25 | 2009-09-09 | Genentech Inc | Nouvelles compositions et methodes de traitement du psoriasis |
EP2272958A1 (en) | 2002-09-26 | 2011-01-12 | ISIS Pharmaceuticals, Inc. | Modulation of forkhead box O1A expression |
EP2298805A3 (en) | 2002-09-27 | 2011-04-13 | Xencor, Inc. | Optimized Fc variants and methods for their generation |
US8129330B2 (en) * | 2002-09-30 | 2012-03-06 | Mountain View Pharmaceuticals, Inc. | Polymer conjugates with decreased antigenicity, methods of preparation and uses thereof |
US20040062748A1 (en) * | 2002-09-30 | 2004-04-01 | Mountain View Pharmaceuticals, Inc. | Polymer conjugates with decreased antigenicity, methods of preparation and uses thereof |
US20060147450A1 (en) * | 2002-10-04 | 2006-07-06 | Shelton David L | Methods for treating cardiac arrhythmia and preventing death due to cardiac arrhythmia using ngf antagonists |
US7432351B1 (en) | 2002-10-04 | 2008-10-07 | Mayo Foundation For Medical Education And Research | B7-H1 variants |
US7255860B2 (en) | 2002-10-08 | 2007-08-14 | Rinat Neuroscience Corp. | Methods for treating post-surgical pain by administering an anti-nerve growth factor antagonist antibody |
UA80447C2 (en) | 2002-10-08 | 2007-09-25 | Methods for treating pain by administering nerve growth factor antagonist and opioid analgesic | |
WO2004032870A2 (en) * | 2002-10-08 | 2004-04-22 | Rinat Neuroscience Corp. | Methods for treating post-surgical pain by admisnistering a nerve growth factor antagonist and compositions containing the same |
US20040146512A1 (en) * | 2002-10-09 | 2004-07-29 | Arnon Rosenthal | Methods of treating Alzheimer's disease using antibodies directed against amyloid beta peptide and compositions thereof |
AU2003304203A1 (en) * | 2002-10-29 | 2005-01-04 | Pharmacia Corporation | Differentially expressed genes involved in cancer, the polypeptides encoded thereby, and methods of using the same |
CA2503330A1 (en) | 2002-10-29 | 2004-05-13 | Genentech, Inc. | Compositions and methods for the treatment of immune related diseases |
WO2004044138A2 (en) | 2002-11-05 | 2004-05-27 | Isis Pharmaceuticals, Inc. | Chimeric oligomeric compounds and their use in gene modulation |
CA2504929C (en) | 2002-11-05 | 2014-07-22 | Charles Allerson | Compositions comprising alternating 2'-modified nucleosides for use in gene modulation |
US20040093198A1 (en) * | 2002-11-08 | 2004-05-13 | Carbon Design Systems | Hardware simulation with access restrictions |
EP1581169A4 (en) | 2002-11-08 | 2008-09-17 | Genentech Inc | Compositions and methods for the treatment of natural killer cell related diseases |
WO2004043396A2 (en) * | 2002-11-09 | 2004-05-27 | Nobex Corporation | Modified carbamate-containing prodrugs and methods of synthesizing same |
DK1569695T3 (en) | 2002-11-13 | 2013-08-05 | Genzyme Corp | ANTISENSE MODULATION OF APOLIPOPROTEIN-B EXPRESSION |
JP4986109B2 (en) | 2002-11-13 | 2012-07-25 | ジェンザイム・コーポレーション | Antisense regulation of apolipoprotein B expression |
US20050158375A1 (en) * | 2002-11-15 | 2005-07-21 | Toshikiro Kimura | Pharmaceutical composition containing liposomes for treating cancer |
US7144999B2 (en) | 2002-11-23 | 2006-12-05 | Isis Pharmaceuticals, Inc. | Modulation of hypoxia-inducible factor 1 alpha expression |
US7648962B2 (en) * | 2002-11-26 | 2010-01-19 | Biocon Limited | Natriuretic compounds, conjugates, and uses thereof |
EP2179742A1 (en) | 2002-11-26 | 2010-04-28 | Genentech, Inc. | Compositions and methods for the treatment of immune related diseases |
EP1569683B1 (en) | 2002-11-26 | 2010-03-03 | Biocon Limited | Modified natriuretic compounds, conjugates, and uses thereof |
US7491699B2 (en) | 2002-12-09 | 2009-02-17 | Ramot At Tel Aviv University Ltd. | Peptide nanostructures and methods of generating and using the same |
EP1587484A4 (en) * | 2002-12-19 | 2008-11-12 | Alza Corp | Method of treating angiogenic tissue growth |
US9498530B2 (en) | 2002-12-24 | 2016-11-22 | Rinat Neuroscience Corp. | Methods for treating osteoarthritis pain by administering a nerve growth factor antagonist and compositions containing the same |
PT1575517E (en) | 2002-12-24 | 2012-05-28 | Rinat Neuroscience Corp | Anti-ngf antibodies and methods using same |
US7569364B2 (en) * | 2002-12-24 | 2009-08-04 | Pfizer Inc. | Anti-NGF antibodies and methods using same |
PT1435231E (en) | 2002-12-31 | 2010-04-08 | Zydus Bsv Pharma Private Ltd | Non-pegylated long-circulating liposomes |
US8980310B2 (en) * | 2002-12-31 | 2015-03-17 | Bharat Serums and Vaccines, Ltd. | Non-pegylated long-circulating liposomes |
ATE426575T1 (en) | 2003-01-07 | 2009-04-15 | Univ Ramot | PEPTIDE ANOSTRUCTURES CONTAINING FOREIGN MATERIAL AND METHOD FOR PRODUCING THE SAME |
WO2004063216A1 (en) * | 2003-01-10 | 2004-07-29 | Yamanouchi Pharmaceutical Co., Ltd. | Conjugate for retention in blood and cancer tissue-specific drug delivery |
US7169892B2 (en) | 2003-01-10 | 2007-01-30 | Astellas Pharma Inc. | Lipid-peptide-polymer conjugates for long blood circulation and tumor specific drug delivery systems |
DK1597366T3 (en) | 2003-02-11 | 2013-02-25 | Antisense Therapeutics Ltd | Modulation of expression of insulin-like growth factor receptor I |
US7655231B2 (en) * | 2003-02-19 | 2010-02-02 | Pfizer Inc. | Methods for treating pain by administering a nerve growth factor antagonist and an NSAID |
US7803781B2 (en) | 2003-02-28 | 2010-09-28 | Isis Pharmaceuticals, Inc. | Modulation of growth hormone receptor expression and insulin-like growth factor expression |
US7968115B2 (en) | 2004-03-05 | 2011-06-28 | Board Of Regents, The University Of Texas System | Liposomal curcumin for treatment of cancer |
WO2004087097A2 (en) * | 2003-03-31 | 2004-10-14 | Yissum Research Development Company Of The Hebrew University Of Jerusalem | Stable liposomes or micelles comprising a sphinolipid and a peg-lipopolymer |
US20040185559A1 (en) | 2003-03-21 | 2004-09-23 | Isis Pharmaceuticals Inc. | Modulation of diacylglycerol acyltransferase 1 expression |
JP2007524604A (en) | 2003-03-31 | 2007-08-30 | アルザ・コーポレーシヨン | Lipid particles having an asymmetric lipid coating and method for producing the same |
PT2335725T (en) | 2003-04-04 | 2017-01-06 | Novartis Ag | High concentration antibody and protein formulations |
ZA200507805B (en) | 2003-04-09 | 2006-12-27 | Genentech Inc | Therapy of autoimmune disease in a patient with an inadequate response to a TNF-alpha inhibitor |
US20070167359A1 (en) * | 2003-04-15 | 2007-07-19 | Moshe Baru | Pharmaceutical composition comprising proteins and/or polypeptides and colloidal particles |
US7598227B2 (en) | 2003-04-16 | 2009-10-06 | Isis Pharmaceuticals Inc. | Modulation of apolipoprotein C-III expression |
WO2004094457A2 (en) * | 2003-04-16 | 2004-11-04 | Arizona Board Of Regents, Acting For And On Behalf Of, Arizona State University | Stable rgd peptidomimetic composition |
EP1620112A4 (en) * | 2003-04-17 | 2007-04-25 | Univ Columbia | Desmoglein 4 is a novel gene involved in hair growth |
US7399853B2 (en) | 2003-04-28 | 2008-07-15 | Isis Pharmaceuticals | Modulation of glucagon receptor expression |
BRPI0411166A (en) | 2003-05-12 | 2006-07-18 | Affymax Inc | compound comprising a peptide moiety, a spacer moiety and a water soluble polymer moiety and a pharmaceutical composition comprising such a compound |
AU2004238869B2 (en) | 2003-05-12 | 2009-06-25 | Affymax, Inc. | Novel poly(ethylene glycol) modified compounds and uses thereof |
JP2007525466A (en) | 2003-05-30 | 2007-09-06 | ジェネンテック・インコーポレーテッド | Treatment with anti-VEGF antibody |
CA2526080A1 (en) * | 2003-05-30 | 2005-01-06 | Genentech, Inc. | Polypeptides that bind an anti-tissue factor antibody and uses thereof |
AU2004253455B2 (en) | 2003-06-03 | 2011-03-03 | Eli Lilly And Company | Modulation of survivin expression |
JP2007526447A (en) | 2003-06-06 | 2007-09-13 | ジェネンテック・インコーポレーテッド | Regulation of interaction between HGF β chain and C-MET |
AU2004247737B2 (en) | 2003-06-11 | 2009-04-23 | Wyeth | Platelet glycoprotein IB alpha variant fusion polypeptides and methods of use thereof |
DE602004028837D1 (en) | 2003-07-03 | 2010-10-07 | Univ New Jersey Med | GENES AS DIAGNOSTIC TOOLS FOR AUTISM |
US7939058B2 (en) * | 2003-07-03 | 2011-05-10 | University Of Southern California | Uses of IL-12 in hematopoiesis |
PL1641822T3 (en) | 2003-07-08 | 2013-10-31 | Genentech Inc | Il-17 a/f heterologous polypeptides and therapeutic uses thereof |
JP4951338B2 (en) * | 2003-07-16 | 2012-06-13 | プロチバ バイオセラピューティクス インコーポレイティッド | Interfering RNA encapsulated in lipid |
CN100431525C (en) * | 2003-07-17 | 2008-11-12 | 台湾东洋药品工业股份有限公司 | Production method of liposome suspended liquid and products thereof |
US20050058699A1 (en) * | 2003-07-31 | 2005-03-17 | The Board Of Regents Of The University Of Texas System | Sterile preparations of phospholipids and anti-inflammatory pharmaceuticals and methods for making and using same |
CA2533701A1 (en) | 2003-07-31 | 2005-02-17 | Isis Pharmaceuticals, Inc. | Oligomeric compounds and compositions for use in modulation of small non-coding rnas |
WO2005019258A2 (en) | 2003-08-11 | 2005-03-03 | Genentech, Inc. | Compositions and methods for the treatment of immune related diseases |
WO2005016348A1 (en) * | 2003-08-14 | 2005-02-24 | Icos Corporation | Method of inhibiting immune responses stimulated by an endogenous factor |
WO2005016349A1 (en) * | 2003-08-14 | 2005-02-24 | Icos Corporation | Methods of inhibiting leukocyte accumulation |
US7825235B2 (en) | 2003-08-18 | 2010-11-02 | Isis Pharmaceuticals, Inc. | Modulation of diacylglycerol acyltransferase 2 expression |
ATE422880T1 (en) * | 2003-08-26 | 2009-03-15 | Smithkline Beecham Corp | HETEROFUNCTIONAL COPOLYMERS OF GLYCEROL AND POLYETHYLENE GLYCOL, THEIR CONJUGATES AND COMPOSITIONS |
US8986731B2 (en) * | 2003-08-26 | 2015-03-24 | Biolitec Pharma Marketing Ltd | Pegylated liposomal formulations of hydrophobic photosensitizers for photodynamic therapy |
US11324698B2 (en) | 2003-08-28 | 2022-05-10 | Vgsk Technologies, Inc. | Sterically stabilized carrier for aerosol therapeutics, compositions and methods for treating the respiratory tract of a mammal |
US20060115523A1 (en) * | 2004-12-01 | 2006-06-01 | Konduri Kameswari S | Sterically stabilized liposome and triamcinolone composition for treating the respiratory tract of a mammal |
US8846079B1 (en) | 2004-12-01 | 2014-09-30 | Vgsk Technologies, Inc. | Sterically stabilized carrier for aerosol therapeutics, compositions and methods for treating the respiratory tract of a mammal |
CN1867580B (en) * | 2003-09-03 | 2010-09-29 | 协和发酵麒麟株式会社 | Compound modified with glycerol derivative |
US20050233435A1 (en) * | 2003-09-04 | 2005-10-20 | Nyu Medical Center | Plasmodium axenic liver stages as a noninfectious whole organism malaria vaccine |
US20070123480A1 (en) * | 2003-09-11 | 2007-05-31 | Replicor Inc. | Oligonucleotides targeting prion diseases |
AU2004272646B2 (en) * | 2003-09-15 | 2011-11-24 | Arbutus Biopharma Corporation | Polyethyleneglycol-modified lipid compounds and uses thereof |
SG146682A1 (en) | 2003-09-18 | 2008-10-30 | Isis Pharmaceuticals Inc | Modulation of eif4e expression |
EP1663199B1 (en) * | 2003-09-25 | 2013-04-03 | Tel Aviv University Future Technology Development L.P. | Compositions and methods using same for treating amyloid-associated diseases |
US8101720B2 (en) * | 2004-10-21 | 2012-01-24 | Xencor, Inc. | Immunoglobulin insertions, deletions and substitutions |
WO2005031362A2 (en) * | 2003-10-02 | 2005-04-07 | Ramot At Tel Aviv University Ltd. | Novel antibacterial agents and methods of identifying and utilizing same |
US7662929B2 (en) | 2003-10-10 | 2010-02-16 | Alchemia Oncology Pty Limited | Antibody that specifically binds hyaluronan synthase |
WO2005034979A2 (en) * | 2003-10-11 | 2005-04-21 | Inex Pharmaceuticals Corporation | Methods and compositions for enhancing innate immunity and antibody dependent cellular cytotoxicity |
TW200517114A (en) | 2003-10-15 | 2005-06-01 | Combinatorx Inc | Methods and reagents for the treatment of immunoinflammatory disorders |
CA2542804A1 (en) * | 2003-10-24 | 2005-05-06 | Alza Corporation | Preparation of lipid particles |
US7960350B2 (en) * | 2003-10-24 | 2011-06-14 | Ader Enterprises, Inc. | Composition and method for the treatment of eye disease |
WO2005040163A1 (en) * | 2003-10-28 | 2005-05-06 | Dr. Reddy's Laboratories Ltd | Heterocyclic compounds that block the effects of advanced glycation end products (age) |
US20050191653A1 (en) | 2003-11-03 | 2005-09-01 | Freier Susan M. | Modulation of SGLT2 expression |
US20050129753A1 (en) * | 2003-11-14 | 2005-06-16 | Gabizon Alberto A. | Method for drug loading in liposomes |
WO2005046637A2 (en) * | 2003-11-14 | 2005-05-26 | Het Nederlands Kanker Instituut | Pharmaceutical formulations employing short-chain sphingolipids and their use |
CA2747871C (en) | 2003-11-17 | 2018-04-10 | Genentech, Inc. | Compositions and methods for the treatment of tumor of hematopoietic origin |
CN1914226B (en) * | 2003-11-25 | 2012-02-01 | 达纳-法伯癌症研究院有限公司 | Antibodies against SARS-COV and methods of use thereof |
US20050142133A1 (en) * | 2003-12-03 | 2005-06-30 | Xencor, Inc. | Optimized proteins that target the epidermal growth factor receptor |
US9050378B2 (en) * | 2003-12-10 | 2015-06-09 | Board Of Regents, The University Of Texas System | N2S2 chelate-targeting ligand conjugates |
US7312320B2 (en) | 2003-12-10 | 2007-12-25 | Novimmune Sa | Neutralizing antibodies and methods of use thereof |
CA2548282A1 (en) | 2003-12-11 | 2005-06-30 | Genentech, Inc. | Methods and compositions for inhibiting c-met dimerization and activation |
CA2551097A1 (en) | 2003-12-23 | 2005-07-14 | Rinat Neuroscience Corp. | Agonist anti-trkc antibodies and methods using same |
PT2311873T (en) | 2004-01-07 | 2018-11-20 | Novartis Vaccines & Diagnostics Inc | M-csf-specific monoclonal antibody and uses thereof |
JP2007520481A (en) * | 2004-01-15 | 2007-07-26 | アルザ・コーポレーシヨン | Liposome composition for delivering therapeutic agents |
WO2005071080A2 (en) | 2004-01-20 | 2005-08-04 | Isis Pharmaceuticals, Inc. | Modulation of glucocorticoid receptor expression |
US7468431B2 (en) | 2004-01-22 | 2008-12-23 | Isis Pharmaceuticals, Inc. | Modulation of eIF4E-BP2 expression |
CA2559853A1 (en) * | 2004-02-17 | 2005-10-13 | University Of South Florida | Materials and methods for treatment of inflammatory and cell proliferation disorders |
US20050181035A1 (en) * | 2004-02-17 | 2005-08-18 | Dow Steven W. | Systemic immune activation method using non CpG nucleic acids |
ATE452147T1 (en) | 2004-02-19 | 2010-01-15 | Genentech Inc | ANTIBODIES WITH CORRECTED CDR |
US8784881B2 (en) | 2004-03-05 | 2014-07-22 | Board Of Regents, The University Of Texas System | Liposomal curcumin for treatment of diseases |
US8569474B2 (en) | 2004-03-09 | 2013-10-29 | Isis Pharmaceuticals, Inc. | Double stranded constructs comprising one or more short strands hybridized to a longer strand |
EP2365077B1 (en) | 2004-03-12 | 2013-05-08 | Alnylam Pharmaceuticals, Inc. | iRNA agents targeting VEGF |
EP1579850A3 (en) * | 2004-03-15 | 2009-12-16 | Nipro Corporation | A pharmaceutical composition containing liposomes for treating a cancer |
EP2700720A3 (en) | 2004-03-15 | 2015-01-28 | Isis Pharmaceuticals, Inc. | Compositions and methods for optimizing cleavage of RNA by RNASE H |
US20070065522A1 (en) * | 2004-03-18 | 2007-03-22 | Transave, Inc. | Administration of high potency platinum compound formulations by inhalation |
US20050249822A1 (en) * | 2004-03-18 | 2005-11-10 | Transave, Inc. | Administration of cisplatin by inhalation |
CA2561221C (en) * | 2004-03-26 | 2016-09-20 | Curis, Inc. | Rna interference modulators of hedgehog signaling and uses thereof |
EP1731172B1 (en) * | 2004-03-26 | 2013-06-05 | Terumo Kabushiki Kaisha | Liposome preparation |
US20050244869A1 (en) * | 2004-04-05 | 2005-11-03 | Brown-Driver Vickie L | Modulation of transthyretin expression |
US7794713B2 (en) | 2004-04-07 | 2010-09-14 | Lpath, Inc. | Compositions and methods for the treatment and prevention of hyperproliferative diseases |
CA2562024C (en) | 2004-04-07 | 2014-05-27 | Rinat Neuroscience Corp. | Methods for treating bone cancer pain by administering a nerve growth factor antagonist |
US20060002930A1 (en) * | 2004-04-16 | 2006-01-05 | Genentech, Inc. | Treatment of disorders |
US20150017671A1 (en) | 2004-04-16 | 2015-01-15 | Yaping Shou | Methods for detecting lp-pla2 activity and inhibition of lp-pla2 activity |
KR101462819B1 (en) | 2004-05-03 | 2014-11-21 | 헤르메스 바이오사이언스, 인코포레이티드 | Liposomes useful for drug delivery |
US8658203B2 (en) | 2004-05-03 | 2014-02-25 | Merrimack Pharmaceuticals, Inc. | Liposomes useful for drug delivery to the brain |
RS55551B1 (en) | 2004-05-13 | 2017-05-31 | Icos Corp | Quinazolinones as inhibitors of human phosphatidylinositol 3-kinase delta |
WO2005120461A2 (en) * | 2004-05-17 | 2005-12-22 | Inex Pharmaceuticals Corporation | Liposomal formulations comprising dihydrosphingomyelin and methods of use thereof |
EP1789453A2 (en) * | 2004-05-18 | 2007-05-30 | Genentech, Inc. | M13 virus major coat protein variants for c-terminal and bi-terminal display of a heterologous protein |
US20050260260A1 (en) * | 2004-05-19 | 2005-11-24 | Edward Kisak | Liposome compositions for the delivery of macromolecules |
WO2005112957A1 (en) * | 2004-05-21 | 2005-12-01 | Transave, Inc. | Treatment of lung diseases and pre-lung disease conditions |
WO2005115383A2 (en) * | 2004-05-25 | 2005-12-08 | Metabolex, Inc. | Substituted triazoles as modulators of ppar and methods of their preparation |
US20060014785A1 (en) * | 2004-05-25 | 2006-01-19 | Metabolex, Inc. | Bicyclic, substituted triazoles as modulators of ppar and methods of their preparation |
JP2008500338A (en) * | 2004-05-25 | 2008-01-10 | イコス・コーポレイション | Method for treating and / or preventing abnormal proliferation of hematopoietic cells |
JP5266492B2 (en) * | 2004-05-28 | 2013-08-21 | ヒューマン バイオモレキュラル リサーチ インスティテュート | Metabolic stable analgesics, pain pharmacotherapy and synthesis of other substances |
AU2005252663B2 (en) * | 2004-06-03 | 2011-07-07 | Isis Pharmaceuticals, Inc. | Double strand compositions comprising differentially modified strands for use in gene modulation |
JP2008501335A (en) * | 2004-06-03 | 2008-01-24 | アイシス ファーマシューティカルズ、インク. | Chimeric gapped oligomer composition |
US8394947B2 (en) | 2004-06-03 | 2013-03-12 | Isis Pharmaceuticals, Inc. | Positionally modified siRNA constructs |
US20090048192A1 (en) * | 2004-06-03 | 2009-02-19 | Isis Pharmaceuticals, Inc. | Double Strand Compositions Comprising Differentially Modified Strands for Use in Gene Modulation |
WO2005117978A2 (en) | 2004-06-04 | 2005-12-15 | Genentech, Inc. | Method for treating multiple sclerosis |
ATE537263T1 (en) | 2004-06-07 | 2011-12-15 | Protiva Biotherapeutics Inc | CATIONIC LIPIDS AND METHODS OF USE |
EP1766035B1 (en) * | 2004-06-07 | 2011-12-07 | Protiva Biotherapeutics Inc. | Lipid encapsulated interfering rna |
EP1771435A4 (en) * | 2004-06-19 | 2008-02-13 | Human Biomolecular Res Inst | Modulators of central nervous system neurotransmitters |
ES2660765T3 (en) | 2004-06-21 | 2018-03-26 | The Board Of Trustees Of The Leland Stanford Junior University | Genes and routes expressed differentially in bipolar disorder and / or major depressive disorder |
WO2006014253A2 (en) * | 2004-07-02 | 2006-02-09 | Genentech, Inc. | Factor viia variants |
JP2008504827A (en) | 2004-07-02 | 2008-02-21 | プロチバ バイオセラピューティクス インコーポレイティッド | Immunostimulatory siRNA molecules and methods of use thereof |
US8143380B2 (en) * | 2004-07-08 | 2012-03-27 | Amgen Inc. | Therapeutic peptides |
WO2006006172A2 (en) * | 2004-07-15 | 2006-01-19 | Ramot At Tel Aviv University Ltd. | Use of anti-amyloid agents for treating and typing pathogen infections |
ES2530340T3 (en) | 2004-07-15 | 2015-03-02 | Xencor Inc | Optimized Fc variants |
WO2006007712A1 (en) * | 2004-07-19 | 2006-01-26 | Protiva Biotherapeutics, Inc. | Methods comprising polyethylene glycol-lipid conjugates for delivery of therapeutic agents |
RU2393168C2 (en) * | 2004-07-19 | 2010-06-27 | Биокон Лимитед | Insulin-oligomer conjugates, preparations and use thereof |
CN101044164A (en) | 2004-07-20 | 2007-09-26 | 健泰科生物技术公司 | Inhibitors of angiopoietin-like 4 protein, combinations, and their use |
WO2006014928A1 (en) | 2004-07-26 | 2006-02-09 | Genentech, Inc. | Methods and compositions for modulating hepatocyte growth factor activation |
AP2007003890A0 (en) * | 2004-07-30 | 2007-02-28 | Rinat Neuroscience Corp | Antibodies directed against amy-loid-beta peptide and methods using same |
US9132116B2 (en) * | 2004-08-02 | 2015-09-15 | Willowcroft Pharm Inc. | Mast cell stabilizers to prevent or treat laminitis |
EP1781310B1 (en) | 2004-08-02 | 2015-10-14 | Ramot at Tel Aviv University Ltd. | Articles of peptide nanostructures and method of forming the same |
EP1778144B1 (en) | 2004-08-17 | 2011-01-19 | Tyco Healthcare Group LP | Anti-adhesion barrier |
JP2006056807A (en) * | 2004-08-18 | 2006-03-02 | Konica Minolta Medical & Graphic Inc | Preparation for use in photodynamic therapy |
ATE539745T1 (en) | 2004-08-19 | 2012-01-15 | Univ Tel Aviv Future Tech Dev | COMPOSITIONS FOR TREATING AMYLOID-ASSOCIATED DISEASES |
JP4715133B2 (en) * | 2004-08-26 | 2011-07-06 | コニカミノルタエムジー株式会社 | Anti-tumor liposome preparation and production method thereof |
JP2006069929A (en) * | 2004-08-31 | 2006-03-16 | Konica Minolta Medical & Graphic Inc | Preparation for treating mycosis and method for producing the same |
US7884086B2 (en) | 2004-09-08 | 2011-02-08 | Isis Pharmaceuticals, Inc. | Conjugates for use in hepatocyte free uptake assays |
WO2006027780A2 (en) | 2004-09-08 | 2006-03-16 | Ramot At Tel Aviv University Ltd. | Peptide nanostructures containing end-capping modified peptides and methods of generating and using the same |
WO2006027785A1 (en) * | 2004-09-09 | 2006-03-16 | Yissum Research Development Company Of The Hebrew University Of Jerusalem | Liposomal formulations comprising an amphipathic weak base like tempamine for treatment of neurodegenerative conditions |
CN101043875B (en) * | 2004-09-09 | 2014-05-07 | 耶路撒冷希伯来大学伊萨姆研发公司 | Liposomal compositions of glucocorticoid and glucocorticoid derivatives |
US6998028B1 (en) * | 2004-09-24 | 2006-02-14 | Superpower, Inc. | Methods for forming superconducting conductors |
BRPI0516011A (en) * | 2004-09-24 | 2008-08-19 | Amgen Inc | modified fc molecules |
US8435948B2 (en) | 2004-09-29 | 2013-05-07 | Mount Sinai School Of Medicine Of New York University | Methods for inhibiting osteoclastic bone resorption and bone loss comprising administration of an anti-FSH or anti-FSHR antibody |
EP1812060A2 (en) | 2004-10-05 | 2007-08-01 | Genentech, Inc. | Method for treating vasculitis |
ATE514776T1 (en) | 2004-10-05 | 2011-07-15 | California Inst Of Techn | APTAMER-REGULATED NUCLEIC ACIDS AND USES THEREOF |
JP5303146B2 (en) | 2004-10-06 | 2013-10-02 | メイヨ・ファウンデーション・フォー・メディカル・エデュケーション・アンド・リサーチ | B7-H1 and methods for diagnosis, prognosis and treatment of cancer |
EP1809246B1 (en) * | 2004-10-08 | 2008-07-16 | Alza Corporation | Method of insertion of a lipid-linked moiety into a pre-formed lipid assembly using microwaves |
TW200612993A (en) * | 2004-10-08 | 2006-05-01 | Alza Corp | Lipopolymer conjugates |
JO3000B1 (en) | 2004-10-20 | 2016-09-05 | Genentech Inc | Antibody Formulations. |
SI1827500T1 (en) * | 2004-10-26 | 2009-10-31 | Pharma Mar Sa | Pegylated liposomal doxorubicin in combination with ecteinescidin 743 |
JP2008518951A (en) * | 2004-10-28 | 2008-06-05 | アルザ コーポレイション | Lyophilized liposome formulations and methods |
DK1658848T3 (en) | 2004-10-29 | 2007-11-26 | Pharma Mar Sa | Formulations comprising ecteinascidin and a disaccharide |
AU2005304914B2 (en) | 2004-11-05 | 2012-02-16 | Tekmira Pharmaceuticals Corporation | Compositions and methods for stabilizing liposomal camptothecin formulations |
TW200618820A (en) * | 2004-11-05 | 2006-06-16 | Alza Corp | Liposome formulations of boronic acid compounds |
US20060246124A1 (en) * | 2004-11-08 | 2006-11-02 | Pilkiewicz Frank G | Methods of treating cancer with lipid-based platinum compound formulations administered intraperitoneally |
AU2005310189A1 (en) * | 2004-11-11 | 2006-06-08 | Affymax, Inc. | Novel peptides that bind to the erythropoietin receptor |
EP1817004A2 (en) * | 2004-11-15 | 2007-08-15 | Yissum Research Development Company Of The Hebrew University Of Jerusalem | Combination therapy associating preferably a ceramide with a cytotoxic drug |
WO2006053430A1 (en) * | 2004-11-17 | 2006-05-26 | Protiva Biotherapeutics, Inc. | Sirna silencing of apolipoprotein b |
WO2006054589A1 (en) | 2004-11-18 | 2006-05-26 | Terumo Kabushiki Kaisha | Medicinal composition, medicinal preparation, and combination preparation |
AU2005316384B2 (en) * | 2004-12-14 | 2012-02-09 | Alnylam Pharmaceuticals, Inc. | RNAi modulation of MLL-AF4 and uses thereof |
ES2343746T3 (en) | 2005-01-07 | 2010-08-09 | Diadexus, Inc. | OVR110 ANTIBODY COMPOSITIONS AND METHODS OF USE. |
US20080207505A1 (en) * | 2005-01-12 | 2008-08-28 | James Kenneth D | Bna Conjugates and Methods of Use |
RU2438705C2 (en) | 2005-01-21 | 2012-01-10 | Дженентек, Инк. | Introduction of fixed doses of her-antibodies |
JP5221126B2 (en) * | 2005-01-28 | 2013-06-26 | 協和発酵キリン株式会社 | Method for producing fine particles surface-modified with water-soluble substances |
CA2596079A1 (en) | 2005-01-31 | 2006-08-10 | Vaxinnate Corporation | Novel polypeptide ligands for toll-like receptor 2 (tlr2) |
US8029783B2 (en) | 2005-02-02 | 2011-10-04 | Genentech, Inc. | DR5 antibodies and articles of manufacture containing same |
CA2596506C (en) | 2005-02-09 | 2021-04-06 | Avi Biopharma, Inc. | Antisense composition and method for treating muscle atrophy |
CA2597924C (en) | 2005-02-15 | 2018-10-02 | Duke University | Anti-cd19 antibodies and uses in oncology |
EP1885356A2 (en) * | 2005-02-17 | 2008-02-13 | Icos Corporation | Phosphoinositide 3-kinase inhibitors for inhibiting leukocyte accumulation |
WO2006089141A2 (en) | 2005-02-18 | 2006-08-24 | Dana-Farber Cancer Institute | Antibodies against cxcr4 and methods of use thereof |
CN103027893B (en) * | 2005-02-18 | 2014-11-12 | 国立大学法人德岛大学 | Polyoxyalkylene chain-containing lipid derivative and lipid film structure containing such derivative |
NZ590431A (en) | 2005-02-23 | 2012-08-31 | Genentech Inc | Extending time to disease progression or survival in cancer patients using a HER dimerization inhibitor |
TW200714289A (en) * | 2005-02-28 | 2007-04-16 | Genentech Inc | Treatment of bone disorders |
WO2006096861A2 (en) * | 2005-03-08 | 2006-09-14 | Genentech, Inc. | METHODS FOR IDENTIFYING TUMORS RESPONSIVE TO TREATMENT WITH HER DIMERIZATION INHIBITORS (HDIs) |
JP2006248978A (en) * | 2005-03-10 | 2006-09-21 | Mebiopharm Co Ltd | New liposome preparation |
RU2007137489A (en) | 2005-03-10 | 2009-04-20 | Дженентек, Инк. (Us) | METHODS AND COMPOSITIONS FOR MODULATION OF VESSEL INTEGRITY |
WO2006099445A2 (en) * | 2005-03-14 | 2006-09-21 | Massachusetts Institute Of Technology | Nanocells for diagnosis and treatment of diseases and disorders |
US8288438B2 (en) | 2005-03-21 | 2012-10-16 | Metabolex, Inc. | Methods for avoiding edema in the treatment or prevention of PPARγ-responsive diseases, including cancer |
CN1840193B (en) * | 2005-03-29 | 2010-05-12 | 中国科学院生物物理研究所 | Nanometer capsule of anthracene nucleus anticancer antibiotic with polyethylene glycol-phospholipid |
ATE541928T1 (en) | 2005-03-31 | 2012-02-15 | Calando Pharmaceuticals Inc | RIBONUCLEOTIDE REDUCTASE SUBUNITY 2 INHIBITORS AND USES THEREOF |
MY148086A (en) * | 2005-04-29 | 2013-02-28 | Rinat Neuroscience Corp | Antibodies directed against amyloid-beta peptide and methods using same |
AU2006244445B2 (en) | 2005-05-05 | 2013-04-18 | Duke University | Anti-CD19 antibody therapy for autoimmune disease |
BRPI0611069A2 (en) | 2005-05-06 | 2010-11-09 | Zymogenetics Inc | monoclonal antibody, humanized antibody, antibody fragment, use of an antagonist, and hybridoma |
EP1899364B1 (en) | 2005-05-17 | 2020-02-19 | University of Connecticut | Compositions and methods for immunomodulation in an organism |
US7919461B2 (en) * | 2005-06-03 | 2011-04-05 | Affymax, Inc. | Erythropoietin receptor peptide formulations and uses |
US8324159B2 (en) * | 2005-06-03 | 2012-12-04 | Affymax, Inc. | Erythropoietin receptor peptide formulations and uses |
US7550433B2 (en) | 2005-06-03 | 2009-06-23 | Affymax, Inc. | Erythropoietin receptor peptide formulations and uses |
CA2610709A1 (en) | 2005-06-06 | 2006-12-14 | Genentech, Inc. | Transgenic models for different genes and their use for gene characterization |
TW200726485A (en) * | 2005-07-01 | 2007-07-16 | Alza Corp | Liposomal delivery vehicle for hydrophobic drugs |
CA2652434A1 (en) | 2005-07-08 | 2007-01-18 | Xencor, Inc. | Optimized proteins that target ep-cam |
CA2615615A1 (en) | 2005-07-22 | 2007-02-01 | Y's Therapeutics Co., Ltd. | Anti-cd26 antibodies and methods of use thereof |
US20080233118A1 (en) * | 2005-07-28 | 2008-09-25 | Novartis Ag | Uses Of Antibody To M-Csf |
JP5457671B2 (en) * | 2005-07-28 | 2014-04-02 | ノバルティス アーゲー | M-CSF specific monoclonal antibody and use thereof |
US20070055200A1 (en) | 2005-08-10 | 2007-03-08 | Gilbert Scott J | Needle-free jet injection drug delivery device |
US8008453B2 (en) | 2005-08-12 | 2011-08-30 | Amgen Inc. | Modified Fc molecules |
WO2007021423A2 (en) | 2005-08-15 | 2007-02-22 | Genentech, Inc. | Gene disruptions, compositions and methods relating thereto |
US20070054873A1 (en) * | 2005-08-26 | 2007-03-08 | Protiva Biotherapeutics, Inc. | Glucocorticoid modulation of nucleic acid-mediated immune stimulation |
US7700567B2 (en) | 2005-09-29 | 2010-04-20 | Supergen, Inc. | Oligonucleotide analogues incorporating 5-aza-cytosine therein |
EP1973928A2 (en) * | 2005-10-11 | 2008-10-01 | Ramot at Tel-Aviv University Ltd. | Self-assembled fmoc-ff hydrogels |
US7875602B2 (en) * | 2005-10-21 | 2011-01-25 | Sutter West Bay Hospitals | Camptothecin derivatives as chemoradiosensitizing agents |
AR056142A1 (en) | 2005-10-21 | 2007-09-19 | Amgen Inc | METHODS TO GENERATE THE MONOVALENT IGG ANTIBODY |
EP1961065A4 (en) | 2005-10-31 | 2009-11-11 | Oncomed Pharm Inc | Compositions and methods for diagnosing and treating cancer |
GB0522082D0 (en) | 2005-10-31 | 2005-12-07 | Pharma Mar Sa | Formulations |
EP2202239A1 (en) | 2005-11-01 | 2010-06-30 | Alnylam Pharmaceuticals Inc. | RNAI inhibition of influenza virus replication |
WO2007051303A1 (en) * | 2005-11-02 | 2007-05-10 | Protiva Biotherapeutics, Inc. | MODIFIED siRNA MOLECULES AND USES THEREOF |
US7879212B2 (en) * | 2005-11-03 | 2011-02-01 | Ramot At Tel-Aviv University Ltd. | Peptide nanostructure-coated electrodes |
DE102005053066A1 (en) | 2005-11-04 | 2007-05-10 | Basf Ag | Use of copolymers as solubilizers for sparingly water-soluble compounds |
US20070190181A1 (en) * | 2005-11-08 | 2007-08-16 | Pilkiewicz Frank G | Methods of treating cancer with lipid-based platinum compound forumulations administered intravenously |
US20070190180A1 (en) * | 2005-11-08 | 2007-08-16 | Pilkiewicz Frank G | Methods of treating cancer with high potency lipid-based platinum compound formulations administered intravenously |
US20070190182A1 (en) * | 2005-11-08 | 2007-08-16 | Pilkiewicz Frank G | Methods of treating cancer with high potency lipid-based platinum compound formulations administered intraperitoneally |
US9107824B2 (en) | 2005-11-08 | 2015-08-18 | Insmed Incorporated | Methods of treating cancer with high potency lipid-based platinum compound formulations administered intraperitoneally |
JP5366554B2 (en) | 2005-11-12 | 2013-12-11 | ザ ボード オブ トラスティーズ オブ ザ レランド スタンフォード ジュニア ユニバーシティー | FGF2-related methods for diagnosing and treating depression |
ME00419B (en) | 2005-11-14 | 2011-10-10 | Rinat Neuroscience Corp | Antagonist antibodies directed against calcitonin gene-related peptide and methods using same |
TW200736277A (en) | 2005-11-14 | 2007-10-01 | Amgen Inc | RANKL antibody-PTH/PTHrP chimeric molecules |
MY149159A (en) | 2005-11-15 | 2013-07-31 | Hoffmann La Roche | Method for treating joint damage |
AP2809A (en) | 2005-11-18 | 2013-12-31 | Glenmark Pharmaceuticals Sa | Anti-Alpha2 Integrin antibodies and their uses |
JP2009516514A (en) | 2005-11-21 | 2009-04-23 | ジェネンテック・インコーポレーテッド | Novel gene disruptions, compositions and methods related thereto |
CA2630602A1 (en) | 2005-11-21 | 2007-05-31 | Isis Pharmaceuticals, Inc. | Modulation of eif4e-bp2 expression |
WO2007064658A2 (en) * | 2005-11-30 | 2007-06-07 | Transave, Inc. | Safe and effective methods of administering therapeutic agents |
CA2631961A1 (en) | 2005-12-02 | 2007-11-08 | Genentech, Inc. | Compositions and methods for the treatment of diseases and disorders associated with cytokine signaling relating to antibodies that bind to il-22 |
EP1979379B1 (en) * | 2005-12-02 | 2013-09-18 | Dana-Farber Cancer Institute | Carbonic anhydrase ix (g250) antibodies and methods of use thereof |
US20090155345A1 (en) * | 2005-12-08 | 2009-06-18 | Ben Gurion University Of The Negev Research And Development Authority | Methods for affecting liposome composition ultrasound irradiation |
CN101325976A (en) * | 2005-12-09 | 2008-12-17 | 巴斯夫欧洲公司 | Use of polyvinyl lactam-polyalkylene block copolymers as solubilisers for poorly water-soluble compounds |
US9149543B2 (en) | 2005-12-15 | 2015-10-06 | The Trustees Of The University Of Pennsylvania | Methods and models for rapid, widespread delivery of genetic material to the CNS using non-viral, cationic lipid-mediated vectors |
JP5580535B2 (en) | 2006-01-05 | 2014-08-27 | ノバルティス アーゲー | Methods for preventing and treating cancer metastasis and bone loss associated with cancer metastasis |
WO2007082144A2 (en) * | 2006-01-05 | 2007-07-19 | Mayo Foundation For Medical Education And Research | B7-h1 and survivin in cancer |
US20090215084A1 (en) * | 2006-01-05 | 2009-08-27 | Mayo Foundation For Medical Education And Research | B7-h1 and b7-h4 in cancer |
SI2161038T1 (en) | 2006-01-26 | 2014-05-30 | Isis Pharmaceuticals, Inc. | Compositions and their uses directed to Huntingtin |
EP2781524A1 (en) | 2006-01-26 | 2014-09-24 | Recopharma AB | Compositions and methods for inhibiting viral adhesion |
WO2007130725A2 (en) * | 2006-02-06 | 2007-11-15 | University Of Pittsburgh Of The Commonwealth System Of Higher Education | Use of hmgb1 for protection against ischemia reperfusion injury |
ZA200807714B (en) | 2006-02-17 | 2010-01-27 | Genentech Inc | Gene disruptions, compositions and methods relating thereto |
EP2650306A1 (en) | 2006-03-06 | 2013-10-16 | Aeres Biomedical Limited | Humanized Anti-CD22 antibodies and their use in treatment of oncology, transplantation and autoimmune disease |
US20070218116A1 (en) * | 2006-03-14 | 2007-09-20 | Schwendener Reto A | Compositions and methods for the treatment of tumors and tumor metastases |
US9119782B2 (en) * | 2006-03-20 | 2015-09-01 | Mary P. McCourt | Drug delivery means |
EP2366716A3 (en) | 2006-03-21 | 2011-11-16 | Genentech, Inc. | Combinatorial therapy involving alpha5beta1 antagonists |
MX2008012013A (en) | 2006-03-23 | 2008-10-03 | Novartis Ag | Anti-tumor cell antigen antibody therapeutics. |
AU2007233109B2 (en) | 2006-03-31 | 2010-10-14 | Alnylam Pharmaceuticals, Inc. | Compositions and methods for inhibiting expression of Eg5 gene |
EP2614839A3 (en) | 2006-04-05 | 2015-01-28 | Genentech, Inc. | Method for using BOC/CDO to modulate hedgehog signaling |
US8758723B2 (en) | 2006-04-19 | 2014-06-24 | The Board Of Regents Of The University Of Texas System | Compositions and methods for cellular imaging and therapy |
US20090288176A1 (en) | 2006-04-19 | 2009-11-19 | Genentech, Inc. | Novel Gene Disruptions, Compositions and Methods Relating Thereto |
WO2007124361A2 (en) * | 2006-04-20 | 2007-11-01 | Mayo Foundation For Medical Education And Research | Soluble b7-h1 |
WO2007121522A1 (en) | 2006-04-21 | 2007-11-01 | Minister for Primary Industries For And On Behalf Of The State Of New South Wales | Pestivirus species |
EP2051585A4 (en) * | 2006-04-28 | 2010-06-02 | Univ South Florida | Materials and methods for reducing inflammation by inhibition of the atrial natriuretic peptide receptor |
EP2013222B1 (en) | 2006-04-28 | 2013-02-13 | Alnylam Pharmaceuticals Inc. | Compositions and methods for inhibiting expression of a gene from the jc virus |
WO2007127428A2 (en) * | 2006-04-28 | 2007-11-08 | University Of Florida Research Foundation, Inc. | Double-stranded/self-complementary vectors with a truncated cba promoter and methods of gene delivery |
US8697120B2 (en) * | 2006-05-01 | 2014-04-15 | Johns Hopkins University | Method and use of nano-scale devices for reduction of tissue injury in ischemic and reperfusion injury |
CN101437943A (en) * | 2006-05-03 | 2009-05-20 | 波罗的科技发展有限公司 | Antisense agents combining strongly bound base - modified oligonucleotide and artificial nuclease |
US20070264322A1 (en) * | 2006-05-10 | 2007-11-15 | Huang Ken S | Method for making liposomes conjugated with temperature-sensitive ligands |
EP2023813A4 (en) * | 2006-05-15 | 2013-03-13 | Dmitri B Kirpotin | Magnetic microparticles comprising organic substances |
EP2522678A1 (en) | 2006-05-15 | 2012-11-14 | Sea Lane Biotechnologies, LLC | Neutralizing antibodies to influenza viruses |
BRPI0712034A2 (en) | 2006-05-19 | 2012-01-10 | Alnylam Pharmaceuticals Inc | aha rnai modulation and therapeutic uses thereof |
WO2007137163A2 (en) * | 2006-05-19 | 2007-11-29 | Georgia Tech Research Corporation | Abc transporter ligand |
EP2192200B1 (en) | 2006-05-22 | 2012-10-24 | Alnylam Pharmaceuticals, Inc. | Compositions and methods for inhibiting expression of IKK-B gene |
WO2007137301A2 (en) * | 2006-05-23 | 2007-11-29 | Isis Pharmaceuticals, Inc. | Modulation of chrebp expression |
US8598333B2 (en) * | 2006-05-26 | 2013-12-03 | Alnylam Pharmaceuticals, Inc. | SiRNA silencing of genes expressed in cancer |
US7862812B2 (en) | 2006-05-31 | 2011-01-04 | Lpath, Inc. | Methods for decreasing immune response and treating immune conditions |
US7915399B2 (en) * | 2006-06-09 | 2011-03-29 | Protiva Biotherapeutics, Inc. | Modified siRNA molecules and uses thereof |
US7981425B2 (en) | 2006-06-19 | 2011-07-19 | Amgen Inc. | Thrombopoietic compounds |
KR101376634B1 (en) | 2006-06-19 | 2014-03-27 | 더 존스 홉킨스 유니버시티 | Tumor-specific delivery of therapeutic agents via liposomase |
US20080096900A1 (en) | 2006-06-26 | 2008-04-24 | Amgen Inc. | Methods for treating atherosclerosis |
HUE030269T2 (en) | 2006-06-26 | 2017-04-28 | Macrogenics Inc | Fc riib-specific antibodies and methods of use thereof |
JP5072275B2 (en) * | 2006-07-03 | 2012-11-14 | テルモ株式会社 | Method for separating closed vesicles, method for producing preparation and evaluation method |
WO2008054561A2 (en) | 2006-07-11 | 2008-05-08 | University Of Medicine And Dentistry Of New Jersey | Proteins, nucleic acids encoding the same and associated methods of use |
US8324158B2 (en) * | 2006-07-14 | 2012-12-04 | Georgia Tech Research Corporation | Methods for inhibiting CLC-2 channel with GATX2 |
US8343539B2 (en) | 2006-07-14 | 2013-01-01 | Regents Of The University Of Minnesota | Compounds that bind α5β1 integrin and methods of use |
WO2008011473A2 (en) | 2006-07-19 | 2008-01-24 | Isis Pharmaceuticals, Inc. | Compositions and their uses directed to hbxip |
MX2009000696A (en) | 2006-07-19 | 2009-01-30 | Univ Pennsylvania | Wsx-1/p28 as a target for anti-inflammatory responses. |
JP4936312B2 (en) * | 2006-07-20 | 2012-05-23 | 株式会社島津製作所 | Novel amphiphile, drug delivery system and molecular imaging system using the same |
WO2008013918A2 (en) * | 2006-07-26 | 2008-01-31 | Myelin Repair Foundation, Inc. | Cell cycle regulation and differentiation |
EP2420252A1 (en) | 2006-08-04 | 2012-02-22 | Novartis AG | EPHB3-specific antibody and uses thereof |
AU2007284651B2 (en) | 2006-08-09 | 2014-03-20 | Institute For Systems Biology | Organ-specific proteins and methods of their use |
ES2402591T3 (en) | 2006-08-14 | 2013-05-07 | Xencor Inc. | Optimized antibodies that target CD19 |
NZ596834A (en) | 2006-08-18 | 2013-06-28 | Novartis Ag | Prlr-specific antibody and uses thereof |
JP2010501172A (en) | 2006-08-25 | 2010-01-21 | オンコセラピー・サイエンス株式会社 | Prognostic markers and therapeutic targets for lung cancer |
SG170032A1 (en) | 2006-08-28 | 2011-04-29 | Kyowa Hakko Kirin Co Ltd | Antagonistic human light-specific human monoclonal antibodies |
MY163119A (en) | 2006-08-29 | 2017-08-15 | Genentech Inc | Use of tenecteplase for treating acute ischemic stroke |
EP2759549B1 (en) | 2006-09-01 | 2015-08-19 | ZymoGenetics, Inc. | IL-31 monoclonal antibodies and methods of use |
CA2600220C (en) * | 2006-09-07 | 2014-12-09 | Canadian Blood Services | Surface cross-linked lipidic particles, methods of production and uses therefor |
EP3357932A1 (en) | 2006-09-29 | 2018-08-08 | OncoMed Pharmaceuticals, Inc. | Compositions and methods for diagnosing and treating cancer |
EP2099467B1 (en) | 2006-10-03 | 2017-05-10 | University Of Medicine And Dentistry Of New Jersey | Atap peptides, nucleic acids encoding the same and associated methods of use |
US10925977B2 (en) | 2006-10-05 | 2021-02-23 | Ceil>Point, LLC | Efficient synthesis of chelators for nuclear imaging and radiotherapy: compositions and applications |
EP2081602A2 (en) | 2006-10-25 | 2009-07-29 | Amgen Inc. | Toxin peptide therapeutic agents |
WO2008055072A2 (en) | 2006-10-27 | 2008-05-08 | Lpath, Inc. | Compositions and methods for treating ocular diseases and conditions |
ZA200902860B (en) | 2006-10-27 | 2010-07-28 | Lpath Inc | Compositions and methods for binding sphingosine-1-phosphate |
WO2008058291A2 (en) * | 2006-11-09 | 2008-05-15 | California Institute Of Technology | Modular aptamer-regulated ribozymes |
CA2669185A1 (en) * | 2006-11-10 | 2008-05-15 | Dimerix Bioscience Pty Ltd | Thyrotropin releasing hormone receptor-orexin receptor hetero-dimers/-oligomers |
EP2441768A1 (en) | 2006-11-13 | 2012-04-18 | Eli Lilly & Co. | Thienopyrimidinones for treatment of inflammatory disorders and cancers |
JP5391073B2 (en) | 2006-11-27 | 2014-01-15 | ディアデクサス インコーポレーテッド | Ovr110 antibody compositions and methods of use |
WO2008070780A1 (en) | 2006-12-07 | 2008-06-12 | Novartis Ag | Antagonist antibodies against ephb3 |
EP1932517A3 (en) * | 2006-12-11 | 2008-07-16 | Universiteit Utrecht Holding B.V. | Liposomes containing a polyphenol derivative such as caffeic acid and a method of post-loading thereof |
US20100203110A1 (en) * | 2006-12-18 | 2010-08-12 | The Johns Hopkins University | Therapeutics for Cancer Using 3-Bromopyruvate and Other Selective Inhibitors of ATP Production |
US8278415B2 (en) | 2006-12-21 | 2012-10-02 | Centocor, Inc. | Dimeric high affinity EGFR constructs and uses thereof |
WO2008079982A2 (en) | 2006-12-21 | 2008-07-03 | Centocor, Inc. | Liposome composition for targeting egfr receptor |
WO2008079973A2 (en) * | 2006-12-21 | 2008-07-03 | Centocor, Inc. | Egfr binding peptides and uses thereof |
EP2097448A4 (en) | 2006-12-22 | 2010-07-21 | Univ Utah Res Found | Method of detecting ocular diseases and pathologic conditions and treatment of same |
WO2008083169A2 (en) * | 2006-12-26 | 2008-07-10 | The Johns Hopkins University | Compositions and methods for the treatment of immunologic disorders |
US7989173B2 (en) | 2006-12-27 | 2011-08-02 | The Johns Hopkins University | Detection and diagnosis of inflammatory disorders |
US20090142342A1 (en) * | 2006-12-27 | 2009-06-04 | Johns Hopkins University | B7-h4 receptor agonist compositions and methods for treating inflammation and auto-immune diseases |
CA2673659A1 (en) * | 2006-12-27 | 2008-07-10 | The Johns Hopkins University | Methods of detecting and diagnosing inflamatory responses and disorders by determining the level of soluble b7-h4 |
US7638541B2 (en) | 2006-12-28 | 2009-12-29 | Metabolex Inc. | 5-ethyl-2-{4-[4-(4-tetrazol-1-yl-phenoxymethyl)-thiazol-2-yl]-piperidin-1-yl}-pyrimidine |
EP2118118B1 (en) * | 2007-01-19 | 2017-09-27 | Exiqon A/S | Mediated cellular delivery of lna oligonucleotides |
WO2008091655A2 (en) * | 2007-01-23 | 2008-07-31 | The Regents Of The University Of California | Methods, compositions and device for directed and controlled heating and release of agents |
WO2008094945A2 (en) | 2007-01-29 | 2008-08-07 | Isis Pharmaceuticals, Inc. | Compounds and methods for modulating protein expression |
WO2008094275A1 (en) | 2007-01-30 | 2008-08-07 | New York University | Peptides for treatment of conditions associated with nitric oxide |
CN101663407B (en) | 2007-02-22 | 2017-08-08 | 健泰科生物技术公司 | method for detecting inflammatory bowel disease |
NZ578824A (en) | 2007-03-02 | 2012-03-30 | Genentech Inc | Predicting response to a her dimerisation inhibitor based on low her3 expression |
CA2681302C (en) * | 2007-03-19 | 2013-07-23 | Dhiraj Khattar | Proliposomal and liposomal compositions of poorly water-soluble compounds |
JP4510842B2 (en) * | 2007-03-26 | 2010-07-28 | キヤノン株式会社 | Polyhydroxyalkanoate-coated liposome |
JP5350360B2 (en) | 2007-03-29 | 2013-11-27 | アルナイラム ファーマシューティカルズ, インコーポレイテッド | Compositions and methods for inhibiting the expression of genes from Ebola |
AU2008246442B2 (en) | 2007-05-04 | 2014-07-03 | Technophage, Investigacao E Desenvolvimento Em Biotecnologia, Sa | Engineered rabbit antibody variable domains and uses thereof |
WO2008137915A2 (en) | 2007-05-07 | 2008-11-13 | Medimmune, Llc | Anti-icos antibodies and their use in treatment of oncology, transplantation and autoimmune disease |
US20090163698A1 (en) * | 2007-05-11 | 2009-06-25 | John Joseph Grigsby | Method for Preparing Antibody Conjugates |
WO2008141274A1 (en) * | 2007-05-11 | 2008-11-20 | Centocor, Inc. | Anti-alpha v immunoliposome composition, methods and uses |
WO2008141276A1 (en) * | 2007-05-11 | 2008-11-20 | Centocor, Inc. | Anti-alpha-v immunoliposome composition, methods and uses |
US8263081B2 (en) | 2007-05-14 | 2012-09-11 | The University Of Chicago | Antibody-light fusion products for cancer therapeutics |
PT2173381E (en) | 2007-05-14 | 2013-12-13 | Novimmune Sa | Fc receptor-binding polypeptides with modified effector functions |
EP2738257A1 (en) | 2007-05-22 | 2014-06-04 | Amgen Inc. | Compositions and methods for producing bioactive fusion proteins |
WO2008148023A2 (en) * | 2007-05-23 | 2008-12-04 | Medical College Of Georgia Research Institute, Inc. | Compositions and methods for treating neurological disorders |
EP3486653A1 (en) | 2007-05-24 | 2019-05-22 | The United States Government as represented by The Department of Veterans Affairs | Treatment of skeletal muscle disorder using ent2 |
DK2164992T3 (en) * | 2007-05-30 | 2016-08-15 | Lpath Inc | COMPOSITIONS AND METHODS FOR BONDING OF LYTHOPHOSPHATIC ACID |
AU2008260498B2 (en) | 2007-05-30 | 2012-11-29 | Xencor, Inc. | Methods and compositions for inhibiting CD32b expressing cells |
US9163091B2 (en) * | 2007-05-30 | 2015-10-20 | Lpath, Inc. | Compositions and methods for binding lysophosphatidic acid |
AR066984A1 (en) | 2007-06-15 | 2009-09-23 | Novartis Ag | INHIBITION OF THE EXPRESSION OF THE ALFA SUBUNITY OF THE SODIUM EPITELIAL CHANNEL (ENAC) THROUGH ARNI (INTERFERENCE RNA) |
TW200916094A (en) * | 2007-06-27 | 2009-04-16 | Poniard Pharmaceuticals Inc | Stabilized picoplatin dosage form |
EP2170946A2 (en) | 2007-07-13 | 2010-04-07 | The Johns Hopkins University | B7-dc variants |
WO2009011855A2 (en) * | 2007-07-16 | 2009-01-22 | California Institute Of Technology | Selection of nucleic acid-based sensor domains within nucleic acid switch platform |
AR067543A1 (en) | 2007-07-16 | 2009-10-14 | Genentech Inc | ANTI-CD79B ANTIBODIES AND HUMANIZED IMMUNOCATE PLAYERS AND METHODS OF USE |
JP5469600B2 (en) | 2007-07-16 | 2014-04-16 | ジェネンテック, インコーポレイテッド | Anti-CD79b antibody and immunoconjugate and method of use thereof |
US8183381B2 (en) | 2007-07-19 | 2012-05-22 | Metabolex Inc. | N-linked heterocyclic receptor agonists for the treatment of diabetes and metabolic disorders |
US20100204425A1 (en) * | 2007-07-26 | 2010-08-12 | Basf Se | Process for preparing copolymers obtained by graft polymerization in solution and based on polyethers in solid form |
NZ597885A (en) | 2007-07-26 | 2013-08-30 | Amgen Inc | Modified lecithin-cholesterol acyltransferase enzymes |
US7666973B2 (en) | 2007-07-30 | 2010-02-23 | Tyco Healthcare Group Lp | Carbonate copolymers |
US8461303B2 (en) | 2007-08-02 | 2013-06-11 | Gilead Biologics, Inc. | LOX and LOXL2 inhibitors and uses thereof |
JPWO2009020094A1 (en) * | 2007-08-09 | 2010-11-04 | 第一三共株式会社 | Antibodies modified with hydrophobic molecules |
US20090048423A1 (en) * | 2007-08-15 | 2009-02-19 | Tyco Healthcare Group Lp | Phospholipid Copolymers |
US8268958B2 (en) | 2007-08-15 | 2012-09-18 | Tyco Healthcare Group Ip | Phospholipid copolymers |
US8367815B2 (en) * | 2007-08-28 | 2013-02-05 | California Institute Of Technology | Modular polynucleotides for ligand-controlled regulatory systems |
US20120165387A1 (en) | 2007-08-28 | 2012-06-28 | Smolke Christina D | General composition framework for ligand-controlled RNA regulatory systems |
EP2197890A1 (en) * | 2007-09-07 | 2010-06-23 | Gencia Corporation | Mitochondrial compositions and uses thereof |
US8865667B2 (en) * | 2007-09-12 | 2014-10-21 | California Institute Of Technology | Higher-order cellular information processing devices |
US20110014118A1 (en) * | 2007-09-21 | 2011-01-20 | Lawrence Tamarkin | Nanotherapeutic colloidal metal compositions and methods |
KR20100123674A (en) * | 2007-09-21 | 2010-11-24 | 싸이티뮨 사이언스, 인크. | Nanotherapeutic colloidal metal compositions and methods |
US9498493B2 (en) | 2007-09-27 | 2016-11-22 | Immunovaccine Technologies Inc. | Use of liposomes in a carrier comprising a continuous hydrophobic phase for delivery of polynucleotides in vivo |
CN101878229A (en) * | 2007-09-28 | 2010-11-03 | 巴塞尔大学医院 | Immunoliposomes for treatment of cancer |
JP2010539978A (en) | 2007-10-02 | 2010-12-24 | アムジェン インコーポレイテッド | Increased erythropoietin using nucleic acids that can hybridize to micro-RNA and its precursors |
US20100209452A1 (en) * | 2007-10-03 | 2010-08-19 | Immunovaccine Technologies, Inc | Compositions comprising an antigen, an amphipathic compound and a hydrophobic carrier, and uses thereof |
KR101234540B1 (en) * | 2007-10-16 | 2013-02-19 | 바이오콘 리미티드 | An orally administerable solid pharmaceutical composition and a process thereof |
US8361465B2 (en) * | 2007-10-26 | 2013-01-29 | Lpath, Inc. | Use of anti-sphingosine-1-phosphate antibodies in combination with chemotherapeutic agents |
JP2011502502A (en) * | 2007-11-05 | 2011-01-27 | バルティック テクロノジー デヴェロプメント,リミテッド | Use of oligonucleotides containing modified bases in nucleic acid hybridization |
SI2219452T1 (en) | 2007-11-05 | 2016-03-31 | Medimmune, Llc | Methods of treating scleroderma |
ES2662845T3 (en) | 2007-11-07 | 2018-04-10 | Genentech, Inc. | IL-22 for use in the treatment of microbial disorders |
CN102089324B (en) | 2007-11-12 | 2014-04-16 | 特罗科隆科学有限公司 | Compositions and methods for the therapy and diagnosis of influenza |
US20110033476A1 (en) * | 2007-11-12 | 2011-02-10 | Theraclone Sciences Inc. | Compositions and methods for the therapy and diagnosis of influenza |
US8598165B2 (en) | 2007-11-26 | 2013-12-03 | University Of Kansas | Morpholines as selective inhibitors of cytochrome P450 2A13 |
MX2010006148A (en) | 2007-12-06 | 2011-02-23 | Dana Farber Cancer Inst Inc | Antibodies against influenza virus and methods of use thereof. |
ES2570853T3 (en) | 2007-12-07 | 2016-05-20 | Zymogenetics Inc | Humanized antibody molecules specific for IL-31 |
EP2848688A1 (en) | 2007-12-10 | 2015-03-18 | Alnylam Pharmaceuticals, Inc. | Compositions and methods for inhibiting expression of factor VII gene |
US9029524B2 (en) * | 2007-12-10 | 2015-05-12 | California Institute Of Technology | Signal activated RNA interference |
CN102216329A (en) | 2007-12-17 | 2011-10-12 | 辉瑞有限公司 | Treatment of interstitial cystitis |
US8962806B2 (en) | 2007-12-28 | 2015-02-24 | Dana-Farber Cancer Institute, Inc. | Humanized monoclonal antibodies and methods of use |
EP3100718B1 (en) | 2008-01-02 | 2019-11-27 | Arbutus Biopharma Corporation | Improved compositions and methods for the delivery of nucleic acids |
US20090181094A1 (en) * | 2008-01-15 | 2009-07-16 | Eric Yueh-Lang Sheu | Molecular Cage for Sustained Release Control of Pharmaceutical and Cosmetic Agents |
AR070141A1 (en) * | 2008-01-23 | 2010-03-17 | Glenmark Pharmaceuticals Sa | SPECIFIC HUMANIZED ANTIBODIES FOR VON WILLEBRAND FACTOR |
TWI472339B (en) | 2008-01-30 | 2015-02-11 | Genentech Inc | Composition comprising antibody that binds to domain ii of her2 and acidic variants thereof |
PE20091318A1 (en) | 2008-01-31 | 2009-09-16 | Genentech Inc | ANTI-CD79B ANTIBODIES AND IMMUNOCONJUGATES AND METHODS OF USE OF THE SAME |
US20110020325A1 (en) * | 2008-02-29 | 2011-01-27 | Kwon Eugene D | Methods for reducing granulomatous inflammation |
KR101397407B1 (en) | 2008-03-05 | 2014-06-19 | 알닐람 파마슈티칼스 인코포레이티드 | Compositions and methods for inhibiting expression of Eg5 and VEGF genes |
JP5544309B2 (en) | 2008-03-10 | 2014-07-09 | セラクローン サイエンシーズ, インコーポレイテッド | Compositions and methods for treatment and diagnosis of cytomegalovirus infection |
EP2105145A1 (en) * | 2008-03-27 | 2009-09-30 | ETH Zürich | Method for muscle-specific delivery lipid-conjugated oligonucleotides |
CN102046656A (en) | 2008-03-28 | 2011-05-04 | 航道生物技术有限责任公司 | Neutralizing molecules to viral antigens |
US20090270404A1 (en) * | 2008-03-31 | 2009-10-29 | Metabolex, Inc. | Oxymethylene aryl compounds and uses thereof |
EP2283133A2 (en) | 2008-04-04 | 2011-02-16 | Calando Pharmaceuticals, Inc. | Compositions and use of epas1 inhibitors |
CN104655854A (en) | 2008-04-09 | 2015-05-27 | 健泰科生物技术公司 | Novel compositions and methods for the treatment of immune related diseases |
JP2011516078A (en) | 2008-04-10 | 2011-05-26 | セル・シグナリング・テクノロジー・インコーポレイテツド | Compositions and methods for detecting EGFR mutations in cancer |
NZ588583A (en) | 2008-04-15 | 2012-08-31 | Protiva Biotherapeutics Inc | Novel lipid formulations for nucleic acid delivery |
US8324366B2 (en) | 2008-04-29 | 2012-12-04 | Alnylam Pharmaceuticals, Inc. | Compositions and methods for delivering RNAI using lipoproteins |
CA2722668A1 (en) * | 2008-04-29 | 2009-11-05 | Wyeth Llc | Methods for treating inflammation |
AU2009245358A1 (en) * | 2008-05-09 | 2009-11-12 | Recopharma Ab | Compositions and methods for inhibiting shiga toxin and shiga-like toxin |
US20090280134A1 (en) * | 2008-05-09 | 2009-11-12 | Recopharma Ab | Compositions and methods for inhibiting toxin a from clostridium difficile |
SI2279004T1 (en) | 2008-05-16 | 2015-05-29 | F. Hoffmann-La Roche Ag | Use of biomarkers for assessing treatment of gastrointestinal inflammatory disorders with beta7integrin antagonists |
US8093018B2 (en) | 2008-05-20 | 2012-01-10 | Otsuka Pharmaceutical Co., Ltd. | Antibody identifying an antigen-bound antibody and an antigen-unbound antibody, and method for preparing the same |
CN102036658A (en) * | 2008-05-23 | 2011-04-27 | 香港大学 | Combination therapy for the treatment of influenza |
EP2296696B1 (en) * | 2008-06-05 | 2014-08-27 | ImmunoVaccine Technologies Inc. | Compositions comprising liposomes, an antigen, a polynucleotide and a carrier comprising a continuous phase of a hydrophobic substance |
WO2009148121A1 (en) | 2008-06-05 | 2009-12-10 | 株式会社 島津製作所 | Novel molecular assembly, molecular probe for molecular imaging and molecular probe for drug delivery system using the same, and molecular imaging system and drug delivery system |
US8173621B2 (en) * | 2008-06-11 | 2012-05-08 | Gilead Pharmasset Llc | Nucleoside cyclicphosphates |
WO2009150623A1 (en) | 2008-06-13 | 2009-12-17 | Pfizer Inc | Treatment of chronic prostatitis |
US20110104074A1 (en) * | 2008-06-18 | 2011-05-05 | University Of Louisville Research Foundation, Inc. | Methods for targeted cancer treatment and detection |
US20100003315A1 (en) | 2008-07-02 | 2010-01-07 | Willeford Kenneth L | Method and Composition for the Treatment of Skin Conditions |
AU2009269095B2 (en) | 2008-07-08 | 2016-05-19 | Oncomed Pharmaceuticals, Inc. | Notch-binding agents and antagonists and methods of use thereof |
CN102149749B (en) | 2008-07-10 | 2014-06-25 | 塞瑞纳治疗公司 | Polyoxazolines with inert terminating groups, polyoxazolines prepared from protected initiating groups and related compounds |
US20100008900A1 (en) * | 2008-07-14 | 2010-01-14 | The University Of Hong Kong | Annexin ii compositions for treating or monitoring inflammation or immune-mediated disorders |
WO2010008582A2 (en) | 2008-07-18 | 2010-01-21 | Rxi Pharmaceuticals Corporation | Phagocytic cell drug delivery system |
EP2323667A4 (en) * | 2008-08-07 | 2012-07-25 | Isis Pharmaceuticals Inc | Modulation of transthyretin expression for the treatment of cns related disorders |
WO2010019702A2 (en) | 2008-08-12 | 2010-02-18 | Oncomed Pharmaceuticals, Inc. | Ddr1-binding agents and methods of use thereof |
CN102186820B (en) | 2008-08-15 | 2013-08-28 | 乔治城大学 | Fluorescent regulators of rassf1a expression and human cancer cell proliferation |
PL2331092T3 (en) | 2008-08-21 | 2014-08-29 | Univ Johns Hopkins | Methods and compositions for administration of 3-halopyruvate and related compounds for the treatment of cancer |
MX2011001135A (en) * | 2008-08-22 | 2011-03-21 | Sanofi Aventis | [4-(5-aminomethyl-2-fluoro-phenyl)-piperidin-1-yl]-[7-fluoro-1-( 2-methoxy-ethyl)-4-trifluoromethoxy-1h-indol-3-yl]-methan one as an inhibitor of mast cell tryptase. |
MX2011002252A (en) | 2008-08-25 | 2011-06-24 | Amplimmune Inc | Compositions of pd-1 antagonists and methods of use. |
CN104740610A (en) * | 2008-08-25 | 2015-07-01 | 安普利穆尼股份有限公司 | PD-1 Antagonists and Methods for Treating Infectious Disease |
EP2328928A2 (en) | 2008-08-25 | 2011-06-08 | Dana-Farber Cancer Institute, Inc. | Conserved influenza hemagglutinin epitope and antibodies thereto |
JP5420668B2 (en) | 2008-08-25 | 2014-02-19 | エクスカリアード・ファーマシューティカルズ,インコーポレイテッド | Antisense oligonucleotides for connective tissue growth factor and uses thereof |
EP2331690B1 (en) | 2008-09-02 | 2016-01-13 | Alnylam Pharmaceuticals Inc. | Compositions and methods for inhibiting expression of mutant egfr gene |
EP2334702A2 (en) | 2008-09-10 | 2011-06-22 | Genentech, Inc. | Methods for inhibiting ocular angiogenesis |
TWI445716B (en) | 2008-09-12 | 2014-07-21 | Rinat Neuroscience Corp | Pcsk9 antagonists |
AR073295A1 (en) * | 2008-09-16 | 2010-10-28 | Genentech Inc | METHODS TO TREAT PROGRESSIVE MULTIPLE SCLEROSIS. MANUFACTURING ARTICLE. |
CA2753338A1 (en) | 2008-09-22 | 2010-03-25 | Rxi Pharmaceuticals Corporation | Neutral nanotransporters |
EP3109321B1 (en) | 2008-09-25 | 2019-05-01 | Alnylam Pharmaceuticals, Inc. | Lipid formulated compositions and methods for inhibiting expression of serum amyloid a gene |
CA2740000C (en) | 2008-10-09 | 2017-12-12 | Tekmira Pharmaceuticals Corporation | Improved amino lipids and methods for the delivery of nucleic acids |
AU2009302217A1 (en) | 2008-10-09 | 2010-04-15 | Northeastern University | Multifunctional self-assembling polymeric nanosystems |
EP3848461A1 (en) | 2008-10-20 | 2021-07-14 | Alnylam Pharmaceuticals, Inc. | Compositions and methods for inhibiting expression of transthyretin |
AU2009307656B2 (en) | 2008-10-21 | 2015-07-02 | Cymabay Therapeutics, Inc. | Aryl GPR120 receptor agonists and uses thereof |
EP2342217B1 (en) * | 2008-10-21 | 2015-12-23 | International Vaccine Institute | Novel shigella frotein antigens and methods |
US8871202B2 (en) | 2008-10-24 | 2014-10-28 | Lpath, Inc. | Prevention and treatment of pain using antibodies to sphingosine-1-phosphate |
CA2742899A1 (en) * | 2008-11-06 | 2010-05-14 | Glenmark Pharmaceuticals, S.A. | Treatment with anti-alpha2 integrin antibodies |
AU2009313201B2 (en) | 2008-11-10 | 2016-06-16 | Arbutus Biopharma Corporation | Novel lipids and compositions for the delivery of therapeutics |
CN102271683B (en) | 2008-11-13 | 2014-07-09 | 吉里德卡利斯托加公司 | Therapies for hematologic malignancies |
US9492449B2 (en) | 2008-11-13 | 2016-11-15 | Gilead Calistoga Llc | Therapies for hematologic malignancies |
SI2361085T2 (en) | 2008-11-22 | 2018-11-30 | F. Hoffmann-La Roche Ag | Use of anti-vegf antibody in combination with chemotherapy for treating breast cancer |
WO2010068414A2 (en) * | 2008-11-25 | 2010-06-17 | Bowen Richard L | Methods for treating obesity related disease |
US20110160222A1 (en) * | 2008-11-26 | 2011-06-30 | Metabolex, Inc. | Modulators of glucose homeostasis for the treatment of diabetes and metabolic disorders |
KR101829469B1 (en) | 2008-12-04 | 2018-03-30 | 큐알엔에이, 인크. | Treatment of erythropoietin (epo) related diseases by inhibition of natural antisense transcript to epo |
RU2746478C2 (en) | 2008-12-04 | 2021-04-14 | КьюРНА, Инк. | Treatment of tumors of diseases related to the genom-suppressor by therapy of natural transcript inhibition in anti-significant orientation regarding this gene |
WO2010065662A2 (en) | 2008-12-04 | 2010-06-10 | Curna, Inc. | Treatment of sirtuin 1 (sirt1) related diseases by inhibition of natural antisense transcript to sirtuin 1 |
WO2010068680A1 (en) | 2008-12-10 | 2010-06-17 | Mount Sinai School Of Medicine Of New York University | Thelper cell type 17 lineage-specific adjuvants, compositions and methods |
JP5855462B2 (en) | 2008-12-10 | 2016-02-09 | アルナイラム ファーマシューティカルズ, インコーポレイテッドAlnylam Pharmaceuticals, Inc. | DsRNA compositions targeting GNAQ and methods for inhibiting expression |
AR074776A1 (en) | 2008-12-18 | 2011-02-09 | Sanofi Aventis | METHOD TO TREAT MACULAR DEGENERATION; MODULATING THE PATIENT'S IMMUNE SYSTEM |
AR074760A1 (en) | 2008-12-18 | 2011-02-09 | Metabolex Inc | GPR120 RECEIVER AGONISTS AND USES OF THE SAME IN MEDICINES FOR THE TREATMENT OF DIABETES AND METABOLIC SYNDROME. |
EP2381964B1 (en) | 2008-12-22 | 2014-06-25 | Creabilis S.a. | Synthesis of polymer conjugates of indolocarbazole compounds |
WO2010075249A2 (en) | 2008-12-22 | 2010-07-01 | Genentech, Inc. | A method for treating rheumatoid arthritis with b-cell antagonists |
SG172361A1 (en) | 2008-12-23 | 2011-07-28 | Pharmasset Inc | Nucleoside analogs |
EP2376088B1 (en) * | 2008-12-23 | 2017-02-22 | Gilead Pharmasset LLC | 6-O-Substituted-2-amino-purine nucleoside phosphoramidates |
US8716263B2 (en) | 2008-12-23 | 2014-05-06 | Gilead Pharmasset Llc | Synthesis of purine nucleosides |
WO2010078536A1 (en) | 2009-01-05 | 2010-07-08 | Rxi Pharmaceuticals Corporation | Inhibition of pcsk9 through rnai |
CA3036963A1 (en) | 2009-01-29 | 2010-08-05 | Arbutus Biopharma Corporation | Lipid formulations comprising cationic lipid and a targeting lipid comprising n-acetyl galactosamine for delivery of nucleic acid |
WO2010086828A2 (en) | 2009-02-02 | 2010-08-05 | Rinat Neuroscience Corporation | Agonist anti-trkb monoclonal antibodies |
WO2010090762A1 (en) | 2009-02-04 | 2010-08-12 | Rxi Pharmaceuticals Corporation | Rna duplexes with single stranded phosphorothioate nucleotide regions for additional functionality |
KR101682735B1 (en) | 2009-02-12 | 2016-12-06 | 큐알엔에이, 인크. | Treatment of brain derived neurotrophic factor (bdnf) related diseases by inhibition of natural antisense transcript to bdnf |
ES2658626T3 (en) | 2009-02-12 | 2018-03-12 | Curna, Inc. | Treatment of diseases related to glial cell-derived neurotrophic factor (GDNF) by inhibition of natural antisense transcript to GDNF |
US8329882B2 (en) | 2009-02-18 | 2012-12-11 | California Institute Of Technology | Genetic control of mammalian cells with synthetic RNA regulatory systems |
US20120041051A1 (en) | 2009-02-26 | 2012-02-16 | Kevin Fitzgerald | Compositions And Methods For Inhibiting Expression Of MIG-12 Gene |
EP2403863B1 (en) | 2009-03-02 | 2013-08-28 | Alnylam Pharmaceuticals Inc. | Nucleic acid chemical modifications |
JP6250263B2 (en) | 2009-03-04 | 2017-12-20 | クルナ・インコーポレーテッド | Treatment of SIRT1-related diseases by suppression of natural antisense transcripts against sirtuin 1 (SIRT1) |
EP2228059A1 (en) | 2009-03-12 | 2010-09-15 | Universitätsspital Basel | Chemotherapeutic composition for the treatment of cancer |
EP2406376A1 (en) | 2009-03-12 | 2012-01-18 | Alnylam Pharmaceuticals, Inc. | LIPID FORMULATED COMPOSITIONS AND METHODS FOR INHIBITING EXPRESSION OF Eg5 AND VEGF GENES |
ES2656290T3 (en) | 2009-03-16 | 2018-02-26 | Curna, Inc. | Treatment of diseases related to nuclear factor (derived from erythroid 2) similar to 2 (NRF2) by inhibition of natural antisense transcript to NRF2 |
EP2408920B1 (en) | 2009-03-17 | 2017-03-08 | CuRNA, Inc. | Treatment of delta-like 1 homolog (dlk1) related diseases by inhibition of natural antisense transcript to dlk1 |
SI3260136T1 (en) | 2009-03-17 | 2021-05-31 | Theraclone Sciences, Inc. | Human immunodeficiency virus (hiv) -neutralizing antibodies |
MX2011009955A (en) | 2009-03-24 | 2011-11-18 | Gilead Calistoga Llc | Atropisomers of2-purinyl-3-tolyl-quinazolinone derivatives and methods of use. |
BRPI1006270B1 (en) | 2009-03-25 | 2022-08-16 | Genentech, Inc | ANTI-A5SS1 ANTIBODY, IMMUNOCONJUGATE, PHARMACEUTICAL COMPOSITION, IN VITRO OR EX VIVO METHOD TO DETECT A5SS1 PROTEIN, USE OF AN ANTIBODY AND KIT TO DETECT A5SS1 PROTEIN |
US8466260B2 (en) | 2009-04-01 | 2013-06-18 | Genentech, Inc. | Anti-FcRH5 antibodies and immunoconjugates and methods of use |
SG174891A1 (en) | 2009-04-01 | 2011-11-28 | Genentech Inc | Treatment of insulin-resistant disorders |
US9145555B2 (en) | 2009-04-02 | 2015-09-29 | California Institute Of Technology | Integrated—ligand-responsive microRNAs |
KR20120005523A (en) * | 2009-04-20 | 2012-01-16 | 길리아드 칼리스토가 엘엘씨 | Methods of treatment for solid tumors |
EP3524275A1 (en) | 2009-04-22 | 2019-08-14 | Massachusetts Institute Of Technology | Innate immune supression enables repeated delivery of long rna molecules |
US8609101B2 (en) | 2009-04-23 | 2013-12-17 | Theraclone Sciences, Inc. | Granulocyte-macrophage colony-stimulating factor (GM-CSF) neutralizing antibodies |
ES2708124T3 (en) | 2009-04-27 | 2019-04-08 | Oncomed Pharm Inc | Procedure for preparing heteromultimeric molecules |
EP2248903A1 (en) | 2009-04-29 | 2010-11-10 | Universitat Autònoma De Barcelona | Methods and reagents for efficient and targeted gene transfer to monocytes and macrophages |
US8524784B2 (en) | 2009-04-30 | 2013-09-03 | Intezyne Technologies, Incorporated | Polymer micelles containing anthracylines for the treatment of cancer |
WO2010127271A1 (en) | 2009-04-30 | 2010-11-04 | Intezyne Technologies, Incorporated | Polymer micelles containing anthracylines for the treatment of cancer |
EP2424987B1 (en) | 2009-05-01 | 2017-11-15 | CuRNA, Inc. | Treatment of hemoglobin (hbf/hbg) related diseases by inhibition of natural antisense transcript to hbf/hbg |
KR20220150411A (en) | 2009-05-05 | 2022-11-10 | 알닐람 파마슈티칼스 인코포레이티드 | Lipid compositions |
CN102458437B (en) | 2009-05-05 | 2015-06-10 | 诺维莫尼公司 | Anti-il-17f antibodies and methods of use thereof |
CA2760706C (en) | 2009-05-05 | 2019-08-06 | Alnylam Pharmaceuticals, Inc. | Methods of delivering oligonucleotides to immune cells |
JP5883782B2 (en) | 2009-05-06 | 2016-03-15 | クルナ・インコーポレーテッド | Treatment of lipid transport metabolism gene-related diseases by suppression of natural antisense transcripts on lipid transport metabolism genes |
WO2010129746A2 (en) | 2009-05-06 | 2010-11-11 | Curna, Inc. | Treatment of tristetraproline (ttp) related diseases by inhibition of natural antisense transcript to ttp |
DK2432881T3 (en) | 2009-05-18 | 2018-02-26 | Curna Inc | TREATMENT OF REPROGRAMMING FACTOR-RELATED DISEASES BY INHIBITING NATURAL ANTISENSE TRANSCRIPTS TO A REPROGRAMMING FACTOR |
CA2762302A1 (en) | 2009-05-20 | 2010-11-25 | Theraclone Sciences, Inc. | Compositions and methods for the therapy and diagnosis of influenza |
TWI598358B (en) | 2009-05-20 | 2017-09-11 | 基利法瑪席特有限責任公司 | Nucleoside phosphoramidates |
EP2432499A2 (en) | 2009-05-20 | 2012-03-28 | Schering Corporation | Modulation of pilr receptors to treat microbial infections |
KR101703695B1 (en) | 2009-05-22 | 2017-02-08 | 큐알엔에이, 인크. | Treatment of transcription factor e3 (tfe3) and insulin receptor substrate 2 (irs2) related diseases by inhibition of natural antisense transcript to tfe3 |
CN103221541B (en) | 2009-05-28 | 2017-03-01 | 库尔纳公司 | Antiviral gene relevant disease is treated by the natural antisense transcript suppressing antiviral gene |
PL3431076T3 (en) | 2009-06-10 | 2022-01-31 | Arbutus Biopharma Corporation | Improved lipid formulation |
US20120100206A1 (en) | 2009-06-11 | 2012-04-26 | Yissum Research Development Company Of The Hebrew University Of Jerusalem, Ltd. | Targeted liposomes comprising n-containing bisphosphonates and uses thereof |
WO2010148050A2 (en) | 2009-06-16 | 2010-12-23 | Curna, Inc. | Treatment of collagen gene related diseases by inhibition of natural antisense transcript to a collagen gene |
JP6128846B2 (en) | 2009-06-16 | 2017-05-17 | クルナ・インコーポレーテッド | Treatment of PON1 gene-related diseases by suppression of natural antisense transcripts against paraoxonase (PON1) |
WO2010146511A1 (en) | 2009-06-17 | 2010-12-23 | Pfizer Limited | Treatment of overactive bladder |
US8859515B2 (en) | 2009-06-24 | 2014-10-14 | Curna, Inc. | Treatment of tumor necrosis factor receptor 2 (TNFR2) related diseases by inhibition of natural antisense transcript to TNFR2 |
WO2010151808A1 (en) | 2009-06-26 | 2010-12-29 | Sea Lane Biotechnologies, Llc | Expression of surrogate light chains |
EP2446037B1 (en) | 2009-06-26 | 2016-04-20 | CuRNA, Inc. | Treatment of down syndrome gene related diseases by inhibition of natural antisense transcript to a down syndrome gene |
EP2450055B1 (en) | 2009-06-30 | 2018-01-03 | Obshestvo S OgranichennoyOtvetstvennostiu"OncoMax" | Method for suppressing renal tumor growth by blocking fibroblast growth factor receptor |
CA2767127A1 (en) | 2009-07-01 | 2011-01-06 | Protiva Biotherapeutics, Inc. | Novel lipid formulations for delivery of therapeutic agents to solid tumors |
WO2011000106A1 (en) | 2009-07-01 | 2011-01-06 | Protiva Biotherapeutics, Inc. | Improved cationic lipids and methods for the delivery of therapeutic agents |
US9018187B2 (en) | 2009-07-01 | 2015-04-28 | Protiva Biotherapeutics, Inc. | Cationic lipids and methods for the delivery of therapeutic agents |
SG177580A1 (en) | 2009-07-07 | 2012-03-29 | Genentech Inc | Diagnosis and treatment of autoimmune demyelinating diseases |
AU2010272957B2 (en) * | 2009-07-17 | 2016-03-03 | Rigshospitalet | Loading technique for preparing radionuclide and ionophore containing liposomes in which the ionophore is 2-hydroxyquionoline (carbostyril) or structurally related 2-hydroxyquinolines |
CA2768843A1 (en) | 2009-07-21 | 2011-01-27 | Gilead Calistoga Llc | Treatment of liver disorders with pi3k inhibitors |
US9937128B2 (en) | 2009-08-03 | 2018-04-10 | The University Of North Carolina At Chapel Hill | Liposomes comprising a calcium phosphate-containing precipitate |
CA2769665A1 (en) | 2009-08-05 | 2011-02-10 | Opko Curna, Llc | Treatment of insulin gene (ins) related diseases by inhibition of natural antisense transcript to an insulin gene (ins) |
US9029338B2 (en) | 2009-08-14 | 2015-05-12 | Alnylam Pharmaceuticals, Inc. | Lipid formulated compositions and methods for inhibiting expression of a gene from the ebola virus |
WO2011022264A1 (en) | 2009-08-15 | 2011-02-24 | Genentech, Inc. | Anti-angiogenesis therapy for the treatment of previously treated breast cancer |
FR2955324A1 (en) | 2010-01-15 | 2011-07-22 | Sanofi Aventis | DISUBSTITUTED 4- (5-AMINOMETHYL-PHENYL) -PIPERIDIN-1-YL] -1H-INDOL-3-YL] -METHANONES |
CN102482671B (en) | 2009-08-25 | 2017-12-01 | 库尔纳公司 | IQGAP relevant diseases are treated by suppressing the natural antisense transcript of ' gtpase activating protein containing IQ die bodys ' (IQGAP) |
EP2473521A2 (en) | 2009-08-31 | 2012-07-11 | Amplimmune, Inc. | B7-h4 fusion proteins and methods of use thereof |
US20110059111A1 (en) | 2009-09-01 | 2011-03-10 | Los Angeles Biomedical Research Institute At Harbor-Ucla Medical Center | Mammalian receptors as targets for antibody and active vaccination therapy against mold infections |
CA2772715C (en) | 2009-09-02 | 2019-03-26 | Genentech, Inc. | Mutant smoothened and methods of using the same |
CA2775840C (en) | 2009-10-01 | 2018-02-06 | Metabolex, Inc. | Substituted tetrazol-1-yl-phenoxymethyl-thiazol-2-yl-piperidinyl-pyrimidine salts |
CA2780741C (en) | 2009-10-12 | 2023-04-04 | Smith Holdings, Llc | Methods and compositions for modulating gene expression using oligonucleotide based drugs administered in vivo or in vitro |
EP2488643A4 (en) * | 2009-10-15 | 2013-07-03 | Hoffmann La Roche | Chimeric fibroblast growth factors with altered receptor specificity |
PT2488204E (en) | 2009-10-16 | 2016-06-09 | Oncomed Pharm Inc | Therapeutic combination and use of dll4 antagonist antibodies and anti-hypertensive agents |
WO2011049625A1 (en) | 2009-10-20 | 2011-04-28 | Mansour Samadpour | Method for aflatoxin screening of products |
WO2011050194A1 (en) | 2009-10-22 | 2011-04-28 | Genentech, Inc. | Methods and compositions for modulating hepsin activation of macrophage-stimulating protein |
WO2011053468A1 (en) | 2009-10-30 | 2011-05-05 | Sanofi-Aventis | Amino-benzoic acid derivatives for use in the treatment of dihydrogenase-related disorders |
SG10201500895XA (en) | 2009-11-05 | 2015-04-29 | Rhizen Pharmaceuticals Sa | Chromen-4-one Derivatives As Kinase Modulators |
WO2011056234A1 (en) | 2009-11-06 | 2011-05-12 | Fibrogen, Inc. | Treatment for radiation-induced disorders |
JP6174320B2 (en) | 2009-11-17 | 2017-08-02 | エムユーエスシー ファウンデーション フォー リサーチ ディベロップメント | Human monoclonal antibody against human nucleolin |
TWI507524B (en) | 2009-11-30 | 2015-11-11 | Genentech Inc | Compositions and methods for the diagnosis and treatment of tumor |
WO2011071957A1 (en) | 2009-12-07 | 2011-06-16 | Sea Lane Biotechnologies, Llc | Conjugates comprising an antibody surrogate scaffold with improved pharmacokinetic properties |
EP3296398A1 (en) | 2009-12-07 | 2018-03-21 | Arbutus Biopharma Corporation | Compositions for nucleic acid delivery |
AU2010329847A1 (en) | 2009-12-11 | 2012-07-26 | Genecode As | Methods of facilitating neural cell survival using GDNF family ligand (GFL) mimetics or RET signaling pathway activators |
WO2011081904A1 (en) | 2009-12-14 | 2011-07-07 | The United States Of America, As Represented By The Secretary, Department Of Health And Human Services | Delivery of transthyretin across the blood-brain barrier as a treatment for alzheimer's disease |
WO2011084455A2 (en) | 2009-12-16 | 2011-07-14 | Opko Curna, Llc. | Treatment of membrane bound transcription factor peptidase, site 1 (mbtps1) related diseases by inhibition of natural antisense transcript to mbtps1 |
WO2011084357A1 (en) | 2009-12-17 | 2011-07-14 | Schering Corporation | Modulation of pilr to treat immune disorders |
CA2784568A1 (en) | 2009-12-18 | 2011-06-23 | Martin A. Maier | Lipid particles for delivery of nucleic acids |
KR101891352B1 (en) | 2009-12-23 | 2018-08-24 | 큐알엔에이, 인크. | Treatment of hepatocyte growth factor (hgf) related diseases by inhibition of natural antisense transcript to hgf |
RU2012131280A (en) | 2009-12-23 | 2014-02-10 | Санофи | [4- (5-AMINOMETHYL-2-fluorophenyl) piperidin-1-yl] - (1H-pyrrolopyridinyl) methanones and their synthesis |
RU2619185C2 (en) | 2009-12-23 | 2017-05-12 | Курна, Инк. | Treatment of diseases associated with uncoupling proteins 2 (ucp2), by inhibiting of natural antisense transcript to ucp2 |
EP2516468B1 (en) | 2009-12-23 | 2016-03-02 | Synimmune GmbH | Anti-flt3 antibodies and methods of using the same |
BR112012014860A2 (en) | 2009-12-23 | 2016-03-29 | Sanofi Sa | [4- [4- (5-Aminomethyl-2-fluoro-phenyl) -piperidin-1-yl] - (1h-pyrrolo-pyridin-yl) -methanones prodrugs and their synthesis |
EP2519542B1 (en) | 2009-12-28 | 2018-10-10 | OncoTherapy Science, Inc. | Anti-cdh3 antibodies and uses thereof |
EP2519633B1 (en) | 2009-12-29 | 2017-10-25 | CuRNA, Inc. | Treatment of nuclear respiratory factor 1 (nrf1) related diseases by inhibition of natural antisense transcript to nrf1 |
JP5982288B2 (en) | 2009-12-29 | 2016-08-31 | カッパーアールエヌエー,インコーポレイテッド | Treatment of tumor protein 63-related diseases by inhibition of natural antisense transcripts against tumor protein 63 (p63) |
WO2011082187A1 (en) | 2009-12-30 | 2011-07-07 | Genentech, Inc. | Methods for modulating a pdgf-aa mediated biological response |
NO2521784T3 (en) | 2010-01-04 | 2018-05-05 | ||
EP2521785B1 (en) | 2010-01-06 | 2022-03-09 | CuRNA, Inc. | Inhibition of natural antisense transcript to a pancreatic developmental gene for use in a treatment of pancreatic developmental gene related diseases |
US8961929B2 (en) | 2010-01-08 | 2015-02-24 | Fujifilm Corporation | Targeting agent for tumor site |
ES2664866T3 (en) | 2010-01-11 | 2018-04-23 | Curna, Inc. | Treatment of diseases related to sex hormone binding globulin (shbg) by inhibition of the natural antisense transcript to shbg |
TWI535445B (en) | 2010-01-12 | 2016-06-01 | 安可美德藥物股份有限公司 | Wnt antagonists and methods of treatment and screening |
WO2011088215A2 (en) | 2010-01-13 | 2011-07-21 | Oncomed Pharmaceuticals, Inc. | Notch1 binding agents and methods of use thereof |
US8883739B2 (en) | 2010-01-19 | 2014-11-11 | The Trustees Of Columbia University In The City Of New York | Osteocalcin as a treatment for male reproductive disorders |
WO2011089211A1 (en) | 2010-01-22 | 2011-07-28 | Synimmune Gmbh | Anti-cd133 antibodies and methods of using the same |
CA2786568A1 (en) | 2010-01-25 | 2011-07-28 | Curna, Inc. | Treatment of rnase h1 related diseases by inhibition of natural antisense transcript to rnase h1 |
US8198337B2 (en) * | 2010-01-27 | 2012-06-12 | Momentive Performance Materials Inc. | Demulsifier compositions and methods for separating emulsions using the same |
US20130052259A1 (en) | 2010-02-01 | 2013-02-28 | Yechezkel Barenholz | Liposomes comprising amphipathic drugs and method for their preparation |
JP5976548B2 (en) | 2010-02-22 | 2016-08-23 | カッパーアールエヌエー,インコーポレイテッド | Treatment of pyrroline-5-carboxylate reductase 1 (PYCR1) related diseases by inhibition of natural antisense transcripts against PYCR1 |
CA2790866C (en) | 2010-02-23 | 2019-02-12 | Sanofi | Anti-alpha2 integrin antibodies and their uses |
SG183335A1 (en) | 2010-02-23 | 2012-09-27 | Genentech Inc | Compositions and methods for the diagnosis and treatment of tumor |
RU2012140447A (en) | 2010-02-23 | 2014-03-27 | Дженентек, Инк. | ANTIANGIOGENIC THERAPY FOR TREATMENT OF OVARIAN CANCER |
SA114360064B1 (en) | 2010-02-24 | 2016-01-05 | رينات نيوروساينس كوربوريشن | Antagonist anti-il-7 receptor antibodies and methods |
US8889193B2 (en) | 2010-02-25 | 2014-11-18 | The Johns Hopkins University | Sustained delivery of therapeutic agents to an eye compartment |
CN105218674A (en) | 2010-03-11 | 2016-01-06 | 瑞纳神经科学公司 | The antibody combined in pH dependence antigen |
CA3027749A1 (en) | 2010-03-22 | 2011-09-29 | Junyan Ji | Compositions and methods useful for stabilizing protein-containing formulations |
EP2550000A4 (en) | 2010-03-24 | 2014-03-26 | Advirna Inc | Reduced size self-delivering rnai compounds |
AU2011230619C1 (en) | 2010-03-25 | 2016-06-23 | Oregon Health & Science University | CMV glycoproteins and recombinant vectors |
WO2011123468A1 (en) | 2010-03-29 | 2011-10-06 | Alnylam Pharmaceuticals, Inc. | Sirna therapy for transthyretin (ttr) related ocular amyloidosis |
EP2552957A4 (en) | 2010-03-29 | 2013-11-20 | Zymeworks Inc | Antibodies with enhanced or suppressed effector function |
ES2807217T3 (en) | 2010-03-31 | 2021-02-22 | Boehringer Ingelheim Int | Anti-CD40 antibodies |
PL2552930T3 (en) | 2010-03-31 | 2016-02-29 | Gilead Pharmasset Llc | Crystalline (s)-isopropyl 2-(((s)-(((2r,3r,4r,5r)-5-(2,4-dioxo-3,4-dihydropyrimidin-1-(2h)-yl)-4-fluoro-3-hydroxy-4-methyltetrahydrofuran-2-yl)methoxy)(phenoxy)phosphoryl)amino)propanoate |
PL3290428T3 (en) | 2010-03-31 | 2022-02-07 | Gilead Pharmasset Llc | Tablet comprising crystalline (s)-isopropyl 2-(((s)-(((2r,3r,4r,5r)-5-(2,4-dioxo-3,4-dihydropyrimidin-1 (2h)-yl)-4-fluoro-3-hydroxy-4-methyltetrahydrofuran-2-yl)methoxy)(phenoxy)phosphoryl)amino)propanoate |
UY33310A (en) | 2010-03-31 | 2011-10-31 | Pharmasset Inc | ESTEREOSELECTIVE SYNTHESIS OF ASSETS CONTAINING PHOSPHORUS |
US9102938B2 (en) | 2010-04-01 | 2015-08-11 | Alnylam Pharmaceuticals, Inc. | 2′ and 5′ modified monomers and oligonucleotides |
WO2011125015A2 (en) | 2010-04-05 | 2011-10-13 | Bar-Ilan University | Protease-activatable pore-forming polypeptides |
WO2011127337A2 (en) | 2010-04-09 | 2011-10-13 | Opko Curna Llc | Treatment of fibroblast growth factor 21 (fgf21) related diseases by inhibition of natural antisense transcript to fgf21 |
WO2011133871A2 (en) | 2010-04-22 | 2011-10-27 | Alnylam Pharmaceuticals, Inc. | 5'-end derivatives |
WO2011133876A2 (en) | 2010-04-22 | 2011-10-27 | Alnylam Pharmaceuticals, Inc. | Oligonucleotides comprising acyclic and abasic nucleosides and analogs |
WO2011133868A2 (en) | 2010-04-22 | 2011-10-27 | Alnylam Pharmaceuticals, Inc. | Conformationally restricted dinucleotide monomers and oligonucleotides |
US9096660B2 (en) | 2010-04-26 | 2015-08-04 | Abraxis Bioscience, Llc | SPARC binding antibodies and uses thereof |
WO2011135905A1 (en) * | 2010-04-28 | 2011-11-03 | 国立大学法人北海道大学 | Lipid membrane structure |
SI2563920T1 (en) | 2010-04-29 | 2017-05-31 | Ionis Pharmaceuticals, Inc. | Modulation of transthyretin expression |
WO2011139911A2 (en) | 2010-04-29 | 2011-11-10 | Isis Pharmaceuticals, Inc. | Lipid formulated single stranded rna |
CA2797856C (en) | 2010-04-30 | 2018-09-18 | Alexion Pharmaceuticals, Inc. | Anti-c5a antibodies and methods for using the antibodies |
WO2011139387A1 (en) | 2010-05-03 | 2011-11-10 | Opko Curna, Llc | Treatment of sirtuin (sirt) related diseases by inhibition of natural antisense transcript to a sirtuin (sirt) |
SG185027A1 (en) | 2010-05-03 | 2012-11-29 | Genentech Inc | Compositions and methods for the diagnosis and treatment of tumor |
CA2794864A1 (en) | 2010-05-03 | 2011-11-10 | Genentech, Inc. | Compositions and methods useful for reducing the viscosity of protein-containing formulations |
JP5866106B2 (en) | 2010-05-12 | 2016-02-17 | コロンビア ユニヴァーシティ | Method for producing enteroendocrine cells that produce and secrete insulin |
TWI586356B (en) | 2010-05-14 | 2017-06-11 | 可娜公司 | Treatment of par4 related diseases by inhibition of natural antisense transcript to par4 |
DK2571878T3 (en) | 2010-05-17 | 2019-02-11 | Indian Incozen Therapeutics Pvt Ltd | Hitherto unknown 3,5-DISUBSTITUTED-3H-IMIDAZO [4,5-B] PYRIDINE AND 3,5- DISUBSTITUTED -3H- [1,2,3] TRIAZOL [4,5-B] PYRIDINE COMPOUNDS AS MODULATORS OF PROTEIN CHINES |
CA2800348C (en) | 2010-05-18 | 2020-07-21 | Lena A. Basile | Il-12 formulations for enhancing hematopoiesis |
WO2011146568A1 (en) | 2010-05-19 | 2011-11-24 | Genentech, Inc. | Predicting response to a her inhibitor |
RU2585229C2 (en) | 2010-05-26 | 2016-05-27 | Курна, Инк. | Treatment of diseases associated with atonal homolog 1 (aton1) by inhibiting natural antisense transcript of gene aton1 |
CA3102008A1 (en) | 2010-06-02 | 2011-12-08 | Alnylam Pharmaceuticals, Inc. | Compositions and methods directed to treating liver fibrosis |
EP2575882B1 (en) | 2010-06-02 | 2017-11-01 | Dana-Farber Cancer Institute, Inc. | Humanized monoclonal antibodies and methods of use |
WO2011153243A2 (en) | 2010-06-02 | 2011-12-08 | Genentech, Inc. | Anti-angiogenesis therapy for treating gastric cancer |
KR20190039347A (en) | 2010-06-03 | 2019-04-10 | 알닐람 파마슈티칼스 인코포레이티드 | Biodegradable lipids for the delivery of active agents |
US8299117B2 (en) | 2010-06-16 | 2012-10-30 | Metabolex Inc. | GPR120 receptor agonists and uses thereof |
JP5746334B2 (en) | 2010-06-16 | 2015-07-08 | シマベイ セラピューティクス, インコーポレーテッド | GPR120 receptor agonist and use thereof |
CN103153283B (en) * | 2010-06-19 | 2017-05-17 | 健康科学西部大学 | Novel formulation of pegylated-liposome encapsulated glycopeptide antibiotics |
JP5847813B2 (en) | 2010-06-23 | 2016-01-27 | シマベイ セラピューティクス, インコーポレーテッド | Composition of 5-ethyl-2- {4- [4- (4-tetrazol-1-yl-phenoxymethyl) -thiazol-2-yl] -piperidin-1-yl} -pyrimidine |
EP2585045B1 (en) | 2010-06-24 | 2019-08-21 | F.Hoffmann-La Roche Ag | Compositions and methods containing alkylgycosides for stabilizing protein-containing formulations |
CN102309448B (en) * | 2010-06-29 | 2014-07-09 | 中国人民解放军军事医学科学院毒物药物研究所 | Pulmonary delivery ciprofloxacin pharmaceutical composition and preparation method thereof |
US9006417B2 (en) | 2010-06-30 | 2015-04-14 | Protiva Biotherapeutics, Inc. | Non-liposomal systems for nucleic acid delivery |
US20130171241A1 (en) | 2010-07-06 | 2013-07-04 | Novartis Ag | Liposomes with lipids having an advantageous pka-value for rna delivery |
HUE047796T2 (en) | 2010-07-06 | 2020-05-28 | Glaxosmithkline Biologicals Sa | Delivery of rna to trigger multiple immune pathways |
US10487332B2 (en) | 2010-07-06 | 2019-11-26 | Glaxosmithkline Biologicals Sa | Immunisation of large mammals with low doses of RNA |
ES2600892T3 (en) | 2010-07-06 | 2017-02-13 | Glaxosmithkline Biologicals Sa | Virion-like administration particles for self-replicating RNA molecules |
CA2804185C (en) | 2010-07-12 | 2017-03-21 | Covx Technologies Ireland Limited | Multifunctional antibody conjugates |
RU2611190C2 (en) | 2010-07-14 | 2017-02-21 | Курна, Инк. | Treatment of diseases related with gene dlg by inhibition of natural antisense transcript of dlg gene |
AU2011280969A1 (en) | 2010-07-23 | 2013-02-07 | Trustees Of Boston University | Anti-Despr inhibitors as therapeutics for inhibition of pathological angiogenesis and tumor cell invasiveness and for molecular imaging and targeted delivery |
US20130202652A1 (en) | 2010-07-30 | 2013-08-08 | Alnylam Pharmaceuticals, Inc. | Methods and compositions for delivery of active agents |
US20130323269A1 (en) | 2010-07-30 | 2013-12-05 | Muthiah Manoharan | Methods and compositions for delivery of active agents |
WO2012019093A1 (en) | 2010-08-05 | 2012-02-09 | Human Biomolecular Research Institute | Synthetic compounds and methods to decrease nicotine self-administration |
EP2420250A1 (en) | 2010-08-13 | 2012-02-22 | Universitätsklinikum Münster | Anti-Syndecan-4 antibodies |
CA2807664A1 (en) | 2010-08-12 | 2012-02-16 | Theraclone Sciences, Inc. | Anti-hemagglutinin antibody compositions and methods of use thereof |
US9254265B2 (en) | 2010-08-31 | 2016-02-09 | Novartis Ag | Small liposomes for delivery of immunogen encoding RNA |
SI3970742T1 (en) | 2010-08-31 | 2022-08-31 | Glaxosmithkline Biologicals S.A. | Pegylated liposomes for delivery of immunogen-encoding rna |
EP3556396B1 (en) | 2010-08-31 | 2022-04-20 | Theraclone Sciences, Inc. | Human immunodeficiency virus (hiv)-neutralizing antibodies |
US8551479B2 (en) | 2010-09-10 | 2013-10-08 | Oncomed Pharmaceuticals, Inc. | Methods for treating melanoma |
JP5974012B2 (en) | 2010-10-05 | 2016-08-23 | ジェネンテック, インコーポレイテッド | Mutant smoothened and method of using the same |
ES2640755T3 (en) | 2010-10-06 | 2017-11-06 | Curna, Inc. | Treatment of diseases related to Sialidase 4 (neu4) by inhibition of the natural antisense transcript to the neu4 gene |
MX363307B (en) | 2010-10-11 | 2019-03-20 | Novartis Ag Star | Antigen delivery platforms. |
WO2012054723A2 (en) | 2010-10-22 | 2012-04-26 | Opko Curna Llc | Treatment of alpha-l-iduronidase (idua) related diseases by inhibition of natural antisense transcript to idua |
EP3214442A1 (en) | 2010-10-25 | 2017-09-06 | F. Hoffmann-La Roche AG | Treatment of gastrointestinal inflammation and psoriasis and asthmainflammation and psoriasis a |
RU2611195C2 (en) | 2010-10-27 | 2017-02-21 | Курна, Инк. | Treatment of interferon-related developmental regulator 1 (ifrd1) associated diseases by inhibition of natural antisense transcript to ifrd1 |
WO2012061448A1 (en) | 2010-11-04 | 2012-05-10 | Boehringer Ingelheim International Gmbh | Anti-il-23 antibodies |
US20140134181A1 (en) | 2010-11-05 | 2014-05-15 | Kenneth E. Lipson | Treatment Method For Lung Remodeling Diseases |
CN110123830A (en) | 2010-11-09 | 2019-08-16 | 阿尔尼拉姆医药品有限公司 | Composition and method for inhibiting the lipid of the expression of Eg5 and VEGF gene to prepare |
US9518114B2 (en) | 2010-11-12 | 2016-12-13 | Purdue Research Foundation | Treating bladder tumor cells using fibronectin attachment protein as a target |
EP2640831A1 (en) | 2010-11-17 | 2013-09-25 | Sea Lane Biotechnologies,llc. | Influenza virus neutralizing agents that mimic the binding site of an influenza neutralizing antibody |
WO2012068531A2 (en) | 2010-11-18 | 2012-05-24 | The General Hospital Corporation | Novel compositions and uses of anti-hypertension agents for cancer therapy |
CA2818824A1 (en) | 2010-11-23 | 2012-05-31 | Joseph Collard | Treatment of nanog related diseases by inhibition of natural antisense transcript to nanog |
JP6069215B2 (en) | 2010-11-30 | 2017-02-01 | ギリアド ファーマセット エルエルシー | Compound |
WO2012079046A2 (en) | 2010-12-10 | 2012-06-14 | Alnylam Pharmaceuticals, Inc. | Compositions and methods for inhibiting expression of klf-1 and bcl11a genes |
EP2649182A4 (en) | 2010-12-10 | 2015-05-06 | Alnylam Pharmaceuticals Inc | Compositions and methods for increasing erythropoietin (epo) production |
RU2013132610A (en) | 2010-12-14 | 2015-01-20 | Текникл Юнивёсити Оф Денмарк | METHOD FOR PRODUCING NANOPARTICLE COMPOSITION CONTAINING METAL COMPONENTS AND THE COMPOSITION OBTAINED THIS METHOD |
EP2468259A1 (en) * | 2010-12-23 | 2012-06-27 | Traslational Cancer Drugs Pharma, S.L. | Pharmaceutical compositions of pyridinium and quinolinium derivatives |
AU2011353698B2 (en) | 2011-01-05 | 2017-05-11 | Livon Laboratories | Methods of making liposomes, liposome compositions made by the methods, and methods of using the same |
AU2012207606B2 (en) | 2011-01-11 | 2017-02-23 | Alnylam Pharmaceuticals, Inc. | Pegylated lipids and their use for drug delivery |
EP2663304B1 (en) | 2011-01-11 | 2019-11-20 | Dimerix Bioscience Pty Ltd | Combination therapy |
KR101697396B1 (en) | 2011-02-02 | 2017-01-17 | 엑스칼리아드 파마슈티컬즈, 인코포레이티드 | Method of treating keloids or hypertrophic scars using antisense compounds targeting connective tissue growth factor (ctgf) |
RU2440142C1 (en) | 2011-02-07 | 2012-01-20 | Общество С Ограниченной Ответственностью "Онкомакс" | Antibody, stopping or retarding tumour growth (versions), method of suppressing tumour growth, method of diagnosing malignant lesions |
WO2012109495A1 (en) | 2011-02-09 | 2012-08-16 | Metabolic Solutions Development Company, Llc | Cellular targets of thiazolidinediones |
SG192727A1 (en) | 2011-02-14 | 2013-09-30 | Theraclone Sciences Inc | Compositions and methods for the therapy and diagnosis of influenza |
PL226015B1 (en) * | 2011-03-03 | 2017-06-30 | Wrocławskie Centrum Badań Eit + Spółka Z Ograniczoną | Liposome preparation containing anticancer active substance, process for the preparation thereof and pharmaceutical compositions containing thereof |
CN108220297A (en) | 2011-03-09 | 2018-06-29 | 细胞信号科技公司 | For generating the method for monoclonal antibody and reagent |
KR20140012131A (en) | 2011-03-15 | 2014-01-29 | 테라클론 사이언시스, 아이엔씨. | Compositions and methods for the therapy and diagnosis of influenza |
TW201300407A (en) | 2011-03-16 | 2013-01-01 | Amgen Inc | Potent and selective inhibitors of Nav1.3 and Nav1.7 |
US10184942B2 (en) | 2011-03-17 | 2019-01-22 | University Of South Florida | Natriuretic peptide receptor as a biomarker for diagnosis and prognosis of cancer |
US9181308B2 (en) | 2011-03-28 | 2015-11-10 | St. Jude Children's Research Hospital | Methods and compositions employing immunogenic fusion proteins |
SG193923A1 (en) | 2011-03-29 | 2013-11-29 | Alnylam Pharmaceuticals Inc | Compositions and methods for inhibiting expression of tmprss6 gene |
MX2013011130A (en) | 2011-03-31 | 2013-10-30 | Genentech Inc | Methods of administering beta7 integrin antagonists. |
EP2508176A1 (en) | 2011-04-08 | 2012-10-10 | Lipotarg Gmbh | Novel combination treatment of cancer |
WO2012142458A1 (en) | 2011-04-13 | 2012-10-18 | Isis Pharmaceuticals, Inc. | Antisense modulation of ptp1b expression |
EP3403672A1 (en) | 2011-04-20 | 2018-11-21 | Medlmmune, LLC | Antibodies and other molecules that bind b7-h1 and pd-1 |
KR101589135B1 (en) | 2011-04-20 | 2016-01-29 | 주식회사 셀앤바이오 | Humanized anti-EMAPII antibodies and uses thereof |
LT2705029T (en) | 2011-05-04 | 2019-02-11 | Rhizen Pharmaceuticals S.A. | Novel compounds as modulators of protein kinases |
EP2710042A2 (en) | 2011-05-16 | 2014-03-26 | Fabion Pharmaceuticals, Inc. | Multi-specific fab fusion proteins and methods of use |
KR101970634B1 (en) | 2011-06-02 | 2019-04-19 | 프레지던트 앤드 펠로우즈 오브 하바드 칼리지 | Methods and uses for ex vivo tissue culture systems |
ES2653247T3 (en) | 2011-06-09 | 2018-02-06 | Curna, Inc. | Treatment of frataxin-related diseases (FXN) by inhibiting the natural antisense transcript to the FXN gene |
LT2691530T (en) | 2011-06-10 | 2018-08-10 | Oregon Health & Science University | Cmv glycoproteins and recombinant vectors |
CA2839437A1 (en) | 2011-06-16 | 2012-12-20 | Isis Pharmaceuticals, Inc. | Antisense modulation of fibroblast growth factor receptor 4 expression |
EP3388068A1 (en) | 2011-06-21 | 2018-10-17 | Alnylam Pharmaceuticals, Inc. | Composition and methods for inhibition of expression of protein c (proc) genes |
EP3564393A1 (en) | 2011-06-21 | 2019-11-06 | Alnylam Pharmaceuticals, Inc. | Assays and methods for determining activity of a therapeutic agent in a subject |
CA3191066A1 (en) | 2011-06-21 | 2012-12-27 | Alnylam Pharmaceuticals Inc. | Compositions and methods for inhibition of expression of apolipoprotein c-iii (apoc3) genes |
EP2537532A1 (en) | 2011-06-22 | 2012-12-26 | J. Stefan Institute | Cathepsin-binding compounds bound to a nanodevice and their diagnostic and therapeutic use |
EP4134433A1 (en) | 2011-06-23 | 2023-02-15 | Alnylam Pharmaceuticals, Inc. | Serpina1 sirnas: compositions of matter and methods of treatment |
EP2724156B1 (en) | 2011-06-27 | 2017-08-16 | The Jackson Laboratory | Methods and compositions for treatment of cancer and autoimmune disease |
WO2013003652A1 (en) | 2011-06-28 | 2013-01-03 | Sea Lane Biotechnologies, Llc | Multispecific stacked variable domain binding proteins |
CA2840989A1 (en) | 2011-07-06 | 2013-01-10 | Novartis Ag | Immunogenic combination compositions and uses thereof |
HUE051570T2 (en) | 2011-07-13 | 2021-03-01 | Yissum Res Dev Co Of Hebrew Univ Jerusalem Ltd | Liposomes co-encapsulating a bisphosphonate and an amphipathic agent |
US20140161821A1 (en) | 2011-07-14 | 2014-06-12 | Pfizer Inc. | Treatment with anti-pcsk9 antibodies |
WO2013015821A1 (en) | 2011-07-22 | 2013-01-31 | The Research Foundation Of State University Of New York | Antibodies to the b12-transcobalamin receptor |
US9120858B2 (en) | 2011-07-22 | 2015-09-01 | The Research Foundation Of State University Of New York | Antibodies to the B12-transcobalamin receptor |
US20130022551A1 (en) | 2011-07-22 | 2013-01-24 | Trustees Of Boston University | DEspR ANTAGONISTS AND AGONISTS AS THERAPEUTICS |
WO2013019857A2 (en) | 2011-08-01 | 2013-02-07 | Alnylam Pharmaceuticals, Inc. | Method for improving the success rate of hematopoietic stem cell transplants |
EP2756094B1 (en) | 2011-08-15 | 2017-12-27 | Medlmmune, LLC | Anti-b7-h4 antibodies and their uses |
FR2979239A1 (en) * | 2011-08-25 | 2013-03-01 | Trophos | LIPOSOME COMPRISING AT LEAST ONE CHOLESTEROL DERIVATIVE |
US8822651B2 (en) | 2011-08-30 | 2014-09-02 | Theraclone Sciences, Inc. | Human rhinovirus (HRV) antibodies |
PT2751279T (en) | 2011-08-31 | 2017-12-29 | St Jude Children`S Res Hospital | Methods and compositions to detect the level of lysosomal exocytosis activity and methods of use |
MY170725A (en) | 2011-09-09 | 2019-08-27 | Univ Osaka | Dengue-virus serotype neutralizing antibodies |
AU2012216792A1 (en) | 2011-09-12 | 2013-03-28 | International Aids Vaccine Initiative | Immunoselection of recombinant vesicular stomatitis virus expressing HIV-1 proteins by broadly neutralizing antibodies |
WO2013043580A2 (en) | 2011-09-19 | 2013-03-28 | Gencia Corporation | Modified creatine compounds |
CN108410868A (en) | 2011-09-20 | 2018-08-17 | Ionis制药公司 | The antisense of GCGR expression is adjusted |
EP2758438A1 (en) | 2011-09-23 | 2014-07-30 | Amgen Research (Munich) GmbH | Bispecific binding molecules for 5t4 and cd3 |
CA2849409A1 (en) | 2011-09-23 | 2013-03-28 | Technophage, Investigacao E Desenvolvimento Em Biotecnologia, Sa | Anti-tumor necrosis factor-alpha agents and uses thereof |
US8858941B2 (en) | 2011-09-23 | 2014-10-14 | Oncomed Pharmaceuticals, Inc. | VEGF/DLL4 binding agents and uses thereof |
EP2760477B1 (en) | 2011-09-27 | 2018-08-08 | Alnylam Pharmaceuticals, Inc. | Di-aliphatic substituted pegylated lipids |
KR102180667B1 (en) | 2011-09-29 | 2020-11-20 | 피엘엑스 옵코 인코포레이티드 | Ph dependent carriers for targeted release of pharmaceuticals along the gastrointestinal tract, compositions therefrom, and making and using same |
US20130085139A1 (en) | 2011-10-04 | 2013-04-04 | Royal Holloway And Bedford New College | Oligomers |
WO2013049941A1 (en) | 2011-10-06 | 2013-04-11 | Immunovaccine Technologies Inc. | Liposome compositions comprising an adjuvant that activates or increases the activity of tlr2 and uses thereof |
AU2012322618A1 (en) | 2011-10-14 | 2014-05-29 | Genentech, Inc. | Anti-HtrA1 antibodies and methods of use |
US8883989B2 (en) | 2011-10-18 | 2014-11-11 | Regents Of The University Of Minnesota | Fractalkine binding polynucleotides and methods of use |
KR20140084232A (en) | 2011-10-25 | 2014-07-04 | 아이시스 파마수티컬즈 인코포레이티드 | Antisense modulation of gccr expression |
EP2586461A1 (en) | 2011-10-27 | 2013-05-01 | Christopher L. Parks | Viral particles derived from an enveloped virus |
CA2850034A1 (en) | 2011-10-28 | 2013-05-02 | Genentech, Inc. | Therapeutic combinations and methods of treating melanoma |
LT3091029T (en) | 2011-10-31 | 2023-02-27 | F. Hoffmann-La Roche Ag | Anti-il13 antibody formulations |
US10980798B2 (en) | 2011-11-03 | 2021-04-20 | Taiwan Liposome Company, Ltd. | Pharmaceutical compositions of hydrophobic camptothecin derivatives |
US10391056B2 (en) * | 2011-11-03 | 2019-08-27 | Taiwan Lipsome Company, LTD. | Pharmaceutical compositions of hydrophobic camptothecin derivatives |
US8871908B2 (en) | 2011-11-11 | 2014-10-28 | Rinat Neuroscience Corp. | Antibodies specific for Trop-2 and their uses |
TWI679212B (en) | 2011-11-15 | 2019-12-11 | 美商安進股份有限公司 | Binding molecules for e3 of bcma and cd3 |
ES2755732T3 (en) | 2011-11-16 | 2020-04-23 | Boehringer Ingelheim Int | Anti IL-36R antibodies |
CN104284680A (en) | 2011-12-15 | 2015-01-14 | 芝加哥大学 | Methods and compositions for cancer therapy using mutant light molecules with increased affinity to receptors |
WO2013093693A1 (en) | 2011-12-22 | 2013-06-27 | Rinat Neuroscience Corp. | Staphylococcus aureus specific antibodies and uses thereof |
EP2794659A1 (en) | 2011-12-22 | 2014-10-29 | Rinat Neuroscience Corp. | Human growth hormone receptor antagonist antibodies and methods of use thereof |
WO2013093809A1 (en) | 2011-12-23 | 2013-06-27 | Pfizer Inc. | Engineered antibody constant regions for site-specific conjugation and methods and uses therefor |
WO2013101771A2 (en) | 2011-12-30 | 2013-07-04 | Genentech, Inc. | Compositions and method for treating autoimmune diseases |
AR090410A1 (en) | 2012-01-09 | 2014-11-12 | Covx Technologies Ireland Ltd | MUTANT ANTIBODIES AND CONJUGATION OF THE SAME |
US9636381B2 (en) | 2012-01-18 | 2017-05-02 | Neumedicines, Inc. | Methods for radiation protection by administering IL-12 |
AU2013209512B2 (en) | 2012-01-20 | 2017-08-03 | I2 Pharmaceuticals, Inc. | Surrobody cojugates |
US20150004219A1 (en) | 2012-02-02 | 2015-01-01 | Yissum Research Development Company Of The Hebrew University Of Jerusalem, Ltd. | Stable liposomes for drug delivery |
AU2013217114B2 (en) | 2012-02-06 | 2017-03-30 | Inhibrx, Inc. | CD47 antibodies and methods of use thereof |
MX2014009827A (en) * | 2012-02-17 | 2014-09-22 | Celsion Corp | Thermosensitive nanoparticle formulations and method of making the same. |
US9138492B2 (en) * | 2012-02-23 | 2015-09-22 | Canon Kabushiki Kaisha | Particle containing hydrophobic dye having cyanine structure, and contrast agent containing the particle |
CA2864177C (en) | 2012-03-01 | 2019-11-26 | Amgen Research (Munich) Gmbh | Prolonged half-life albumin-binding protein fused bispecific antibodies |
AR090253A1 (en) | 2012-03-05 | 2014-10-29 | Gilead Calistoga Llc | Polymorphic forms of (S) -2- (1- (9H-PURIN-6-ILAMINO) PROPIL) -5-FLUOR-3-PHENYLQUINAZOLIN-4 (3H) -ONA |
CA2867262C (en) | 2012-03-15 | 2021-03-16 | Curna, Inc. | Treatment of brain derived neurotrophic factor (bdnf) related diseases by inhibition of natural antisense transcript to bdnf |
US9453076B2 (en) | 2012-03-29 | 2016-09-27 | Novimmune S.A. | Anti-TLR4 antibodies and uses thereof |
AP3908A (en) | 2012-03-30 | 2016-11-24 | Rhizen Pharmaceuticals Sa | Novel 3,5-disubstituted-3H-imidazo[4,5-B]pyridine and 3,5-disubstituted-3H-[1,2,3]triazolo [4,5-B] pyridine compounds as modulators of C-met protein kinases |
WO2013151649A1 (en) | 2012-04-04 | 2013-10-10 | Sialix Inc | Glycan-interacting compounds |
US9133461B2 (en) | 2012-04-10 | 2015-09-15 | Alnylam Pharmaceuticals, Inc. | Compositions and methods for inhibiting expression of the ALAS1 gene |
KR20150023287A (en) | 2012-04-24 | 2015-03-05 | 더 유니버시티 오브 마이애미 | Perforin 2 defense against invasive and multidrug resistant pathogens |
CN104507497B (en) | 2012-05-03 | 2018-10-19 | 勃林格殷格翰国际有限公司 | Anti-il-23 p 19 antibodies |
CA2871751C (en) | 2012-05-04 | 2021-08-24 | Dana-Farber Cancer Institute, Inc. | Affinity matured anti-ccr4 humanized monoclonal antibodies and methods of use |
JP6392209B2 (en) | 2012-05-04 | 2018-09-19 | ザ・ジョンズ・ホプキンス・ユニバーシティー | Lipid-based drug carriers for rapid permeation through the mucus lining |
US9717724B2 (en) | 2012-06-13 | 2017-08-01 | Ipsen Biopharm Ltd. | Methods for treating pancreatic cancer using combination therapies |
AU2013202947B2 (en) | 2012-06-13 | 2016-06-02 | Ipsen Biopharm Ltd. | Methods for treating pancreatic cancer using combination therapies comprising liposomal irinotecan |
EP2861256B1 (en) | 2012-06-15 | 2019-10-23 | The Brigham and Women's Hospital, Inc. | Compositions for treating cancer and methods for making the same |
WO2013192546A1 (en) | 2012-06-22 | 2013-12-27 | Cytomx Therapeutics, Inc. | Activatable antibodies having non-binding steric moieties and mehtods of using the same |
ES2631608T3 (en) | 2012-06-27 | 2017-09-01 | International Aids Vaccine Initiative | Env-glycoprotein variant of HIV-1 |
JP2015524410A (en) * | 2012-07-18 | 2015-08-24 | オニキス セラピューティクス, インク.Onyx Therapeutics, Inc. | Liposome composition of epoxy ketone-based proteasome inhibitor |
US8603470B1 (en) | 2012-08-07 | 2013-12-10 | National Cheng Kung University | Use of IL-20 antagonists for treating liver diseases |
CN103565745A (en) | 2012-08-10 | 2014-02-12 | 德克萨斯州大学系统董事会 | Neuroprotective liposome compositions and methods for treatment of stroke |
SG11201408340SA (en) | 2012-08-14 | 2015-01-29 | Univ Nanyang Tech | Angiopoietin-like 4 antibody and a method of its use in cancer treatment |
US11291644B2 (en) | 2012-09-04 | 2022-04-05 | Eleison Pharmaceuticals, Llc | Preventing pulmonary recurrence of cancer with lipid-complexed cisplatin |
WO2014041179A1 (en) | 2012-09-17 | 2014-03-20 | Chemedest Ltd. | Treatment of peripheral neuropathy using gfr(alpha)3 type receptor agonists |
CN107892719B (en) | 2012-10-04 | 2022-01-14 | 达纳-法伯癌症研究所公司 | Human monoclonal anti-PD-L1 antibodies and methods of use |
ES2738305T3 (en) | 2012-10-05 | 2020-01-21 | Hoffmann La Roche | Procedures for diagnosing and treating inflammatory bowel diseases |
US9266959B2 (en) | 2012-10-23 | 2016-02-23 | Oncomed Pharmaceuticals, Inc. | Methods of treating neuroendocrine tumors using frizzled-binding agents |
JP6371294B2 (en) | 2012-10-31 | 2018-08-08 | オンコメッド ファーマシューティカルズ インコーポレイテッド | Methods and monitoring of treatment with DLL4 antagonists |
US11059910B2 (en) | 2012-12-03 | 2021-07-13 | Novimmune Sa | Anti-CD47 antibodies and methods of use thereof |
WO2014092858A1 (en) | 2012-12-12 | 2014-06-19 | Temple University - Of The Commonwealth System Of Higher Education | Compositions and methods for treatment of cancer |
EP2934303B1 (en) | 2012-12-19 | 2019-09-04 | The Research Foundation for the State University of New York | Compositions and method for light triggered release of materials from nanovesicles |
AU2013361275B2 (en) | 2012-12-19 | 2016-11-24 | Amplimmune, Inc. | Anti-human B7-H4 antibodies and their uses |
US20160067347A1 (en) | 2012-12-20 | 2016-03-10 | Amgen Inc. | Apj receptor agonists and uses thereof |
GB201223053D0 (en) | 2012-12-20 | 2013-02-06 | Medical Res Council | Receptor |
EP2935332B1 (en) | 2012-12-21 | 2021-11-10 | MedImmune, LLC | Anti-h7cr antibodies |
US9566325B2 (en) | 2013-01-07 | 2017-02-14 | Biomedical Research Models, Inc. | Therapeutic vaccines for treating herpes simplex virus type-2 infections |
ES2728936T3 (en) | 2013-01-25 | 2019-10-29 | Amgen Inc | Antibodies directed against CDH19 for melanoma |
JO3519B1 (en) | 2013-01-25 | 2020-07-05 | Amgen Inc | Antibody constructs for CDH19 and CD3 |
US10568975B2 (en) | 2013-02-05 | 2020-02-25 | The Johns Hopkins University | Nanoparticles for magnetic resonance imaging tracking and methods of making and using thereof |
CA2900468A1 (en) | 2013-02-06 | 2014-08-14 | Inhibrx Llc | Non-platelet depleting and non-red blood cell depleting cd47 antibodies and methods of use thereof |
US9534041B2 (en) | 2013-02-12 | 2017-01-03 | The United States Of America, As Represented By The Secretary, Department Of Health And Human Services | Monoclonal antibodies that neutralize a norovirus |
TW201446792A (en) | 2013-03-12 | 2014-12-16 | Amgen Inc | Potent and selective inhibitors of Nav1.7 |
CA2905108C (en) | 2013-03-14 | 2021-12-07 | Julie HUGHES | Cholestosome vesicles for incorporation of molecules into chylomicrons |
US9302005B2 (en) | 2013-03-14 | 2016-04-05 | Mayo Foundation For Medical Education And Research | Methods and materials for treating cancer |
AR095374A1 (en) | 2013-03-15 | 2015-10-14 | Amgen Res (Munich) Gmbh | UNION MOLECULES FOR BCMA AND CD3 |
WO2014145654A1 (en) | 2013-03-15 | 2014-09-18 | The Trustees Of The University Of Pennsylvania | Method for the site-specific covalent cross-linking of antibodies to surfaces |
BR112015023664A2 (en) | 2013-03-15 | 2017-10-24 | Gencia Corp | compositions and methods for treating conditions affecting the epidermis |
US20140283157A1 (en) | 2013-03-15 | 2014-09-18 | Diadexus, Inc. | Lipoprotein-associated phospholipase a2 antibody compositions and methods of use |
MX367668B (en) | 2013-03-15 | 2019-08-30 | Dana Farber Cancer Inst Inc | Flavivirus neutralizing antibodies and methods of use thereof. |
WO2014140368A1 (en) | 2013-03-15 | 2014-09-18 | Amgen Research (Munich) Gmbh | Antibody constructs for influenza m2 and cd3 |
CA2906624A1 (en) | 2013-03-15 | 2014-09-25 | Dyax Corp. | Anti-plasma kallikrein antibodies |
UY35484A (en) | 2013-03-15 | 2014-10-31 | Amgen Res Munich Gmbh | Single chain binding molecule comprising N-end ABP |
US10052364B2 (en) | 2013-03-15 | 2018-08-21 | The Trustees Of Columbia University In The City Of New York | Osteocalcin as a treatment for cognitive disorders |
CA2907175C (en) * | 2013-03-15 | 2022-11-08 | M. Alphabet 3, L.L.C. | Methods and compositions for enhancing oxygen levels in tissues |
EP3708184A1 (en) | 2013-03-27 | 2020-09-16 | The General Hospital Corporation | Methods and agents for treating alzheimer s disease |
MX2015011765A (en) | 2013-03-27 | 2016-01-15 | Genentech Inc | Use of biomarkers for assessing treatment of gastrointestinal inflammatory disorders with beta7 integrin antagonists. |
US20160067196A1 (en) | 2013-04-12 | 2016-03-10 | Icahn School Of Medicine At Mount Sinai | Method for treating post-traumatic stress disorder |
WO2014179760A1 (en) | 2013-05-03 | 2014-11-06 | The Regents Of The University Of California | Cyclic di-nucleotide induction of type i interferon |
HUE052232T2 (en) | 2013-05-06 | 2021-04-28 | Scholar Rock Inc | Compositions and methods for growth factor modulation |
EP2994167B1 (en) | 2013-05-06 | 2020-05-06 | Alnylam Pharmaceuticals, Inc. | Dosages and methods for delivering lipid formulated nucleic acid molecules |
WO2014181229A2 (en) | 2013-05-07 | 2014-11-13 | Rinat Neuroscience Corp. | Anti-glucagon receptor antibodies and methods of use thereof |
WO2014183052A1 (en) | 2013-05-09 | 2014-11-13 | The United States Of America, As Represented By The Secretary, Depart Of Health And Human Services | Single-domain vhh antibodies directed to norovirus gi.1 and gii.4 and their use |
BR112015029395A2 (en) | 2013-05-24 | 2017-09-19 | Medimmune Llc | ANTI-B7-H5 ANTIBODIES AND THEIR USES |
KR20160015286A (en) | 2013-05-31 | 2016-02-12 | 제넨테크, 인크. | Anti-wall teichoic antibodies and conjugates |
MY177774A (en) | 2013-05-31 | 2020-09-23 | Genentech Inc | Anti-wall teichoic antibodies and conjugates |
NZ714765A (en) | 2013-06-06 | 2021-12-24 | Pf Medicament | Anti-c10orf54 antibodies and uses thereof |
EP3007704B1 (en) | 2013-06-13 | 2021-01-06 | Antisense Therapeutics Ltd | Combination therapy for acromegaly |
US10208125B2 (en) | 2013-07-15 | 2019-02-19 | University of Pittsburgh—of the Commonwealth System of Higher Education | Anti-mucin 1 binding agents and uses thereof |
EP3024851B1 (en) | 2013-07-25 | 2018-05-09 | CytomX Therapeutics, Inc. | Multispecific antibodies, multispecific activatable antibodies and methods of using the same |
WO2015017807A1 (en) * | 2013-08-01 | 2015-02-05 | University Of Georgia Research Foundation, Inc. | Liposomal formulations for the treatment of bacterial infections |
CA2919790C (en) | 2013-08-02 | 2018-06-19 | Pfizer Inc. | Anti-cxcr4 antibodies and antibody-drug conjugates |
AU2014306867B2 (en) | 2013-08-12 | 2017-10-26 | Genentech, Inc. | Compositions and method for treating complement-associated conditions |
WO2015023851A1 (en) | 2013-08-14 | 2015-02-19 | The Governing Council Of The University Of Toronto | Antibodies against frizzled proteins and methods of use thereof |
EP3038600B1 (en) * | 2013-08-27 | 2020-06-03 | Northeastern University | Nanoparticle drug delivery system and method of treating cancer and neurotrauma |
PL3041958T4 (en) | 2013-09-04 | 2020-11-02 | Cold Spring Harbor Laboratory | Reducing nonsense-mediated mrna decay |
US20150065381A1 (en) | 2013-09-05 | 2015-03-05 | International Aids Vaccine Initiative | Methods of identifying novel hiv-1 immunogens |
US11236338B2 (en) | 2013-09-05 | 2022-02-01 | Sarepta Therapeutics, Inc. | Antisense-induced exon2 inclusion in acid alpha-glucosidase |
AR097648A1 (en) | 2013-09-13 | 2016-04-06 | Amgen Inc | COMBINATION OF EPIGENETIC FACTORS AND BIESPECTIVE COMPOUNDS THAT HAVE LIKE DIANA CD33 AND CD3 IN THE TREATMENT OF MYELOID LEUKEMIA |
SI3046537T1 (en) * | 2013-09-16 | 2022-01-31 | Glycomine, Inc. | Pharmaceutical preparation of carbohydrates for therapeutic use |
DE102013015334A1 (en) * | 2013-09-17 | 2015-03-19 | Fresenius Medical Care Deutschland Gmbh | Magnesium-liposome complexes |
EP3047023B1 (en) | 2013-09-19 | 2019-09-04 | The United States Of America, As Represented By The Secretary, Department Of Health And Human Services | Compositions and methods for inhibiting jc virus (jcv) |
CA2925106C (en) | 2013-09-25 | 2023-11-14 | Cytomx Therapeutics, Inc. | Matrix metalloproteinase substrates and other cleavable moieties and methods of use thereof |
WO2015050663A1 (en) | 2013-10-01 | 2015-04-09 | Mayo Foundation For Medical Education And Research | Methods for treating cancer in patients with elevated levels of bim |
EP3052626A1 (en) | 2013-10-02 | 2016-08-10 | Alnylam Pharmaceuticals, Inc. | Compositions and methods for inhibiting expression of the lect2 gene |
TW202310853A (en) | 2013-10-04 | 2023-03-16 | 美國西奈山伊坎醫學院 | Compositions and methods for inhibiting expression of the alas1 gene |
EP2873423B1 (en) | 2013-10-07 | 2017-05-31 | International Aids Vaccine Initiative | Soluble hiv-1 envelope glycoprotein trimers |
WO2015068781A1 (en) | 2013-11-06 | 2015-05-14 | 国立大学法人大阪大学 | Antibody having broad neutralizing activity in group 1 influenza a virus |
WO2015087187A1 (en) | 2013-12-10 | 2015-06-18 | Rinat Neuroscience Corp. | Anti-sclerostin antibodies |
US20160333063A1 (en) | 2013-12-13 | 2016-11-17 | The General Hospital Corporation | Soluble high molecular weight (hmw) tau species and applications thereof |
AU2014364414A1 (en) | 2013-12-20 | 2016-06-30 | Gilead Calistoga Llc | Polymorphic forms of a hydrochloride salt of (S) -2-(1-(9H-purin-6-ylamino) propyl) -5-fluoro-3-phenylquinazolin-4 (3H) -one |
WO2015095601A1 (en) | 2013-12-20 | 2015-06-25 | Gilead Calistoga Llc | Process methods for phosphatidylinositol 3-kinase inhibitors |
CN115054698A (en) | 2014-01-14 | 2022-09-16 | 约翰斯·霍普金斯大学 | Cyclodextrin compositions encapsulating selective ATP inhibitors and uses thereof |
WO2015116902A1 (en) | 2014-01-31 | 2015-08-06 | Genentech, Inc. | G-protein coupled receptors in hedgehog signaling |
US9840543B2 (en) | 2014-01-31 | 2017-12-12 | Boehringer Ingelheim International Gmbh | Anti-BAFF antibodies |
IL294782B2 (en) | 2014-01-31 | 2023-10-01 | Cytomx Therapeutics Inc | Matriptase and u-plasminogen activator polypeptide substrates and other cleanable moieties, compositions comprising same and uses thereof |
AU2015214264B2 (en) | 2014-02-04 | 2018-12-20 | Curis, Inc. | Mutant Smoothened and methods of using the same |
EP3110974A4 (en) | 2014-02-24 | 2018-01-24 | Celgene Corporation | Methods of using an activator of cereblon for neural cell expansion and the treatment of central nervous system disorders |
GB201403775D0 (en) | 2014-03-04 | 2014-04-16 | Kymab Ltd | Antibodies, uses & methods |
NZ631007A (en) | 2014-03-07 | 2015-10-30 | Alexion Pharma Inc | Anti-c5 antibodies having improved pharmacokinetics |
WO2015143194A2 (en) | 2014-03-19 | 2015-09-24 | Dana-Farber Cancer Institute, Inc. | Immunogenetic restriction on elicitation of antibodies |
FI3119431T3 (en) | 2014-03-21 | 2024-03-20 | Teva Pharmaceuticals Int Gmbh | Antagonist antibodies directed against calcitonin gene-related peptide and methods using same |
EP3122377A4 (en) | 2014-03-27 | 2018-03-14 | F.Hoffmann-La Roche Ag | Methods for diagnosing and treating inflammatory bowel disease |
CN106470681A (en) | 2014-04-03 | 2017-03-01 | 茵维特丝肿瘤学私营有限责任公司 | Supramolecular assembly medicine |
EP3134111B1 (en) | 2014-04-25 | 2022-06-08 | Dana-Farber Cancer Institute, Inc. | Middle east respiratory syndrome coronavirus neutralizing antibodies and methods of use thereof |
US10308697B2 (en) | 2014-04-30 | 2019-06-04 | President And Fellows Of Harvard College | Fusion proteins for treating cancer and related methods |
EP3711780A3 (en) | 2014-04-30 | 2020-12-09 | Pfizer Inc | Anti-ptk7 antibody-drug conjugates |
WO2015171918A2 (en) | 2014-05-07 | 2015-11-12 | Louisiana State University And Agricultural And Mechanical College | Compositions and uses for treatment thereof |
JP6868394B2 (en) | 2014-05-16 | 2021-05-12 | ファイザー・インク | Bispecific antibody |
US10302653B2 (en) | 2014-05-22 | 2019-05-28 | Mayo Foundation For Medical Education And Research | Distinguishing antagonistic and agonistic anti B7-H1 antibodies |
EA028614B1 (en) | 2014-05-22 | 2017-12-29 | Общество С Ограниченной Ответственностью "Русские Фармацевтические Технологии" | Selective inhibitors interfering with fibroblast growth factor receptor and frs2 interaction for the prevention and treatment of cancer |
US10584171B2 (en) | 2014-05-30 | 2020-03-10 | Henlix Biotech Co., Ltd. | Anti-epidermal growth factor receptor (EGFR) antibodies |
ES2950789T3 (en) | 2014-06-10 | 2023-10-13 | Amgen Inc | Apelin polypeptides |
EP3155016A1 (en) | 2014-06-11 | 2017-04-19 | Gilead Sciences, Inc. | Methods for treating cardiovascular diseases |
US11021467B2 (en) | 2014-06-13 | 2021-06-01 | Gilead Sciences, Inc. | Phosphatidylinositol 3-kinase inhibitors |
TWI695011B (en) | 2014-06-18 | 2020-06-01 | 美商梅爾莎納醫療公司 | Monoclonal antibodies against her2 epitope and methods of use thereof |
EP3157945A4 (en) | 2014-06-20 | 2017-11-22 | F. Hoffmann-La Roche AG | Chagasin-based scaffold compositions, methods, and uses |
US10487314B2 (en) | 2014-06-26 | 2019-11-26 | The Trustees Of Columbia University In The City Of New York | Inhibition of serotonin expression in gut enteroendocrine cells results in conversion to insulin-positive cells |
EP3171896A4 (en) | 2014-07-23 | 2018-03-21 | Mayo Foundation for Medical Education and Research | Targeting dna-pkcs and b7-h1 to treat cancer |
AU2015292406B2 (en) | 2014-07-25 | 2021-03-11 | Cytomx Therapeutics, Inc | Anti-CD3 antibodies, activatable anti-CD3 antibodies, multispecific anti-CD3 antibodies, multispecific activatable anti-CD3 antibodies, and methods of using the same |
AU2015294834B2 (en) | 2014-07-31 | 2021-04-29 | Amgen Research (Munich) Gmbh | Optimized cross-species specific bispecific single chain antibody constructs |
UY36245A (en) | 2014-07-31 | 2016-01-29 | Amgen Res Munich Gmbh | ANTIBODY CONSTRUCTS FOR CDH19 AND CD3 |
JP2017523980A (en) | 2014-08-06 | 2017-08-24 | ライナット ニューロサイエンス コーポレイション | Method for lowering LDL-cholesterol |
WO2016020799A1 (en) | 2014-08-06 | 2016-02-11 | Rinat Neuroscience Corp. | Methods for reducing ldl-cholesterol |
WO2016033424A1 (en) | 2014-08-29 | 2016-03-03 | Genzyme Corporation | Methods for the prevention and treatment of major adverse cardiovascular events using compounds that modulate apolipoprotein b |
WO2016034968A1 (en) | 2014-09-02 | 2016-03-10 | Pfizer Inc. | Therapeutic antibody |
EP3197492A1 (en) | 2014-09-23 | 2017-08-02 | Pfizer Inc | Treatment with anti-pcsk9 antibodies |
EP3201232A1 (en) | 2014-10-03 | 2017-08-09 | Dana-Farber Cancer Institute, Inc. | Glucocorticoid-induced tumor necrosis factor receptor (gitr) antibodies and methods of use thereof |
GB201417589D0 (en) | 2014-10-06 | 2014-11-19 | Cantab Biopharmaceuticals Patents Ltd | Pharmaceutical Formulations |
CA2962949C (en) | 2014-10-06 | 2024-03-05 | Dana-Farber Cancer Institute, Inc. | Humanized cc chemokine receptor 4 (ccr4) antibodies and methods of use thereof |
MX2017005258A (en) | 2014-10-31 | 2017-07-26 | Oncomed Pharm Inc | Combination therapy for treatment of disease. |
US9879087B2 (en) | 2014-11-12 | 2018-01-30 | Siamab Therapeutics, Inc. | Glycan-interacting compounds and methods of use |
ES2941897T3 (en) | 2014-11-12 | 2023-05-26 | Seagen Inc | Compounds that interact with glycans and procedures for use |
WO2016077566A1 (en) | 2014-11-12 | 2016-05-19 | Research Institute At Nationwide Children's Hospital | Modulation of alternative mdm2 splicing |
JP7175608B2 (en) | 2014-11-19 | 2022-11-21 | ザ トラスティーズ オブ コロンビア ユニバーシティ イン ザ シティ オブ ニューヨーク | Osteocalcin as a treatment for age-related frailty |
KR20170086542A (en) | 2014-12-03 | 2017-07-26 | 제넨테크, 인크. | Anti-staphylococcus aureus antibody rifamycin conjugates and uses thereof |
BR112017011325A2 (en) | 2014-12-03 | 2020-07-21 | Genentech, Inc. | "antibody conjugate compound" antibiotic, composition, method of treating an infection, method of killing staph aureus, process for producing the conjugate, kit for treating an infection, antibiotic-ligand and ligand-drug intermediates |
WO2016090170A1 (en) | 2014-12-05 | 2016-06-09 | The United States Of America, As Represented By The Secretary, Department Of Health And Human Services | A potent anti-influenza a neuraminidase subtype n1 antibody |
TWI595006B (en) | 2014-12-09 | 2017-08-11 | 禮納特神經系統科學公司 | Anti-pd-1 antibodies and methods of use thereof |
CN108064167A (en) | 2014-12-11 | 2018-05-22 | 皮埃尔法布雷医药公司 | Anti- C10ORF54 antibody and application thereof |
TW201702218A (en) | 2014-12-12 | 2017-01-16 | 美國杰克森實驗室 | Compositions and methods relating to the treatment of cancer, autoimmune disease, and neurodegenerative disease |
SG11201704869VA (en) | 2014-12-15 | 2017-07-28 | Univ Johns Hopkins | Sunitinib formulations and methods for use thereof in treatment of ocular disorders |
WO2016100716A1 (en) | 2014-12-18 | 2016-06-23 | Vasant Jadhav | Reversirtm compounds |
CA2974369A1 (en) | 2015-01-20 | 2016-07-28 | The Children's Medical Center Corporation | Anti-net compounds for treating and preventing fibrosis and for facilitating wound healing |
MA41374A (en) | 2015-01-20 | 2017-11-28 | Cytomx Therapeutics Inc | MATRIX METALLOPROTEASE CLIVABLE AND SERINE PROTEASE CLIVABLE SUBSTRATES AND METHODS OF USE THEREOF |
EP3250184A1 (en) | 2015-01-27 | 2017-12-06 | The Johns Hopkins University | Hypotonic hydrogel formulations for enhanced transport of active agents at mucosal surfaces |
CN107407677B (en) | 2015-01-28 | 2020-07-17 | 豪夫迈·罗氏有限公司 | Gene expression markers and treatment of multiple sclerosis |
CN107428818A (en) | 2015-01-29 | 2017-12-01 | 密西根州立大学校董会 | Hide polypeptide and application thereof |
CA2976912A1 (en) * | 2015-02-17 | 2016-08-25 | Mallinckrodt Llc | Modified docetaxel liposome formulations and uses thereof |
US10550173B2 (en) | 2015-02-19 | 2020-02-04 | Compugen, Ltd. | PVRIG polypeptides and methods of treatment |
EP3258951B1 (en) | 2015-02-19 | 2020-01-29 | Compugen Ltd. | Anti-pvrig antibodies and methods of use |
CN107257693A (en) | 2015-02-26 | 2017-10-17 | 豪夫迈·罗氏有限公司 | Treat the integrin beta 7 antagonists and method of Crohn diseases |
ES2937020T3 (en) | 2015-03-03 | 2023-03-23 | Kymab Ltd | Antibodies, uses and methods |
US10736845B2 (en) * | 2015-03-03 | 2020-08-11 | Cureport Inc. | Dual loaded liposomal pharmaceutical formulations |
US9895313B2 (en) | 2015-03-03 | 2018-02-20 | Cureport, Inc. | Combination liposomal pharmaceutical formulations |
EP3268392A2 (en) | 2015-03-13 | 2018-01-17 | CytomX Therapeutics, Inc. | Anti-pdl1 antibodies, activatable anti-pdl1 antibodies, and methods of use thereof |
AU2016233143B2 (en) | 2015-03-17 | 2021-03-25 | Alberto Gabizon | Methods for the treatment of bladder cancer |
MA41795A (en) | 2015-03-18 | 2018-01-23 | Sarepta Therapeutics Inc | EXCLUSION OF AN EXON INDUCED BY ANTISENSE COMPOUNDS IN MYOSTATIN |
US10174292B2 (en) | 2015-03-20 | 2019-01-08 | International Aids Vaccine Initiative | Soluble HIV-1 envelope glycoprotein trimers |
EP3072901A1 (en) | 2015-03-23 | 2016-09-28 | International Aids Vaccine Initiative | Soluble hiv-1 envelope glycoprotein trimers |
US9758575B2 (en) | 2015-04-06 | 2017-09-12 | Yung Shin Pharmaceutical Industrial Co. Ltd. | Antibodies which specifically bind to canine vascular endothelial growth factor and uses thereof |
WO2016164835A1 (en) | 2015-04-08 | 2016-10-13 | Dana-Farber Cancer Institute, Inc. | Humanized influenza monoclonal antibodies and methods of use thereof |
CA3219684A1 (en) | 2015-04-13 | 2016-10-13 | Pfizer Inc. | Anti-bcma antibodies, anti-cd3 antibodies and bi-specific antibodies binding to bcma and cd3 |
TWI772258B (en) | 2015-04-17 | 2022-08-01 | 德商安美基研究(慕尼黑)公司 | Bispecific antibody constructs for cdh3 and cd3 |
CN107849134B (en) | 2015-05-01 | 2022-05-03 | 达纳-法伯癌症研究所公司 | Methods of mediating cytokine expression with anti-CCR 4 antibodies |
IL310251A (en) | 2015-05-04 | 2024-03-01 | Cytomx Therapeutics Inc | Anti-cd71 antibodies, activatable anti-cd71 antibodies, compositions comprising same and uses thereof |
IL292798A (en) | 2015-05-04 | 2022-07-01 | Cytomx Therapeutics Inc | Anti-cd166 antibodies, activatable anti-cd166 antibodies, compositions comprising same and uses thereof |
US10233244B2 (en) | 2015-05-04 | 2019-03-19 | Cytomx Therapeutics, Inc. | Anti-ITGA3 antibodies, activatable anti-ITGA3 antibodies, and methods of use thereof |
EP3294334B1 (en) | 2015-05-11 | 2020-07-08 | The Johns Hopkins University | Autoimmune antibodies for use in inhibiting cancer cell growth |
EP3294887A1 (en) | 2015-05-14 | 2018-03-21 | CNIC Fundación Centro Nacional de Investigaciones Cardiovasculares Carlos III | Mirna compositions for the treatment of mature b-cell neoplasms |
US11318131B2 (en) | 2015-05-18 | 2022-05-03 | Ipsen Biopharm Ltd. | Nanoliposomal irinotecan for use in treating small cell lung cancer |
WO2016196381A1 (en) | 2015-05-29 | 2016-12-08 | Genentech, Inc. | Pd-l1 promoter methylation in cancer |
EP3302497A4 (en) | 2015-06-01 | 2019-01-16 | Sarepta Therapeutics, Inc. | Antisense-induced exon exclusion in type vii collagen |
US11426398B2 (en) | 2015-06-10 | 2022-08-30 | Elysium Health, Inc. | Nicotinamide riboside and pterostilbene compositions and methods for treatment of skin disorders |
US10485764B2 (en) | 2015-07-02 | 2019-11-26 | Otsuka Pharmaceutical Co., Ltd. | Lyophilized pharmaceutical compositions |
CA2991980A1 (en) | 2015-07-13 | 2017-01-19 | Compugen Ltd. | Hide1 compositions and methods |
JP7014706B2 (en) | 2015-07-13 | 2022-02-01 | サイトメックス セラピューティクス インコーポレイテッド | Anti-PD-1 antibody, activating anti-PD-1 antibody, and how to use it |
CA2989970A1 (en) | 2015-07-17 | 2017-01-26 | Alnylam Pharmaceuticals, Inc. | Multi-targeted single entity conjugates |
BR112018001202A2 (en) | 2015-07-21 | 2018-09-25 | Dyax Corp | monoclonal antibody, nucleic acid, vector, host cell, pharmaceutical composition and method |
EP3792279A3 (en) | 2015-07-29 | 2021-07-07 | Allergan, Inc. | Heavy chain only antibodies to ang-2 |
TWI793062B (en) | 2015-07-31 | 2023-02-21 | 德商安美基研究(慕尼黑)公司 | Antibody constructs for dll3 and cd3 |
TWI796283B (en) | 2015-07-31 | 2023-03-21 | 德商安美基研究(慕尼黑)公司 | Antibody constructs for msln and cd3 |
TWI829617B (en) | 2015-07-31 | 2024-01-21 | 德商安美基研究(慕尼黑)公司 | Antibody constructs for flt3 and cd3 |
TWI717375B (en) | 2015-07-31 | 2021-02-01 | 德商安美基研究(慕尼黑)公司 | Antibody constructs for cd70 and cd3 |
TWI744242B (en) | 2015-07-31 | 2021-11-01 | 德商安美基研究(慕尼黑)公司 | Antibody constructs for egfrviii and cd3 |
JP2018523673A (en) | 2015-08-14 | 2018-08-23 | アラーガン、インコーポレイテッドAllergan,Incorporated | Heavy chain only antibody against PDGF |
ES2848118T3 (en) | 2015-08-20 | 2021-08-05 | Ipsen Biopharm Ltd | Combination therapy using liposomal irinotecan and a PARP inhibitor for the treatment of cancer |
BR112018002941B1 (en) | 2015-08-21 | 2023-12-05 | Ipsen Biopharm Ltd | USE OF LIPOSOMAL IRINOTECAN, OXALIPLATIN, LEUCOVORIN AND 5-FLUOROURACIL IN FIRST-LINE TREATMENT OF METASTATIC ADENOCARCINOMA OF THE PANCREAS |
MX2018002447A (en) | 2015-09-01 | 2018-06-15 | Boehringer Ingelheim Int | Use of anti-cd40 antibodies for treatment of lupus nephritis. |
WO2017048843A1 (en) | 2015-09-14 | 2017-03-23 | Alnylam Pharmaceuticals, Inc. | Compositions and methods for inhibiting expression of the alas1 gene |
KR20230155021A (en) | 2015-09-15 | 2023-11-09 | 스칼러 락, 인크. | Anti-pro/latent-myostatin antibodies and uses thereof |
TWI799366B (en) | 2015-09-15 | 2023-04-21 | 美商建南德克公司 | Cystine knot scaffold platform |
JP6967003B2 (en) | 2015-09-23 | 2021-11-17 | メレオ バイオファーマ 5 インコーポレイテッド | Methods and compositions for the treatment of cancer |
WO2017058881A1 (en) | 2015-09-28 | 2017-04-06 | The Trustees Of Columbia University In The City Of New York | Use of pentoxifylline with immune checkpoint-blockade therapies for the treatment of melanoma |
WO2017055966A1 (en) | 2015-10-01 | 2017-04-06 | Pfizer Inc. | Low viscosity antibody compositions |
EP3359572A2 (en) | 2015-10-06 | 2018-08-15 | H. Hoffnabb-La Roche Ag | Method for treating multiple sclerosis |
MA45819A (en) | 2015-10-09 | 2018-08-15 | Sarepta Therapeutics Inc | COMPOSITIONS AND METHODS FOR TREATING DUCHENNE MUSCLE DYSTROPHY AND RELATED DISORDERS |
GB201518170D0 (en) | 2015-10-14 | 2015-11-25 | Cantab Biopharmaceuticals Patents Ltd | Colloidal particles for subcutaneous administration with intravenous administration of therapeutic agent |
GB201518171D0 (en) | 2015-10-14 | 2015-11-25 | Cantab Biopharmaceuticals Patents Ltd | Colloidal particles for topical administration with therapeutic agent |
GB201518172D0 (en) | 2015-10-14 | 2015-11-25 | Cantab Biopharmaceuticals Patents Ltd | Colloidal particles for use in medicine |
MA42991A (en) | 2015-10-16 | 2018-08-22 | Ipsen Biopharm Ltd | STABILIZATION OF PHARMACEUTICAL COMPOSITIONS OF CAMPTOTHECIN |
WO2017066714A1 (en) | 2015-10-16 | 2017-04-20 | Compugen Ltd. | Anti-vsig1 antibodies and drug conjugates |
WO2017075037A1 (en) | 2015-10-27 | 2017-05-04 | Scholar Rock, Inc. | Primed growth factors and uses thereof |
CA3001362C (en) | 2015-10-30 | 2020-10-13 | Genentech, Inc. | Anti-htra1 antibodies and methods of use thereof |
WO2017075045A2 (en) | 2015-10-30 | 2017-05-04 | Mayo Foundation For Medical Education And Research | Antibodies to b7-h1 |
AU2016349954B2 (en) | 2015-11-05 | 2022-08-25 | Antisense Therapeutics Ltd | Mobilizing leukemia cells |
WO2017079563A1 (en) | 2015-11-06 | 2017-05-11 | The Johns Hopkins University | Methods of treating liver fibrosis by administering 3-bromopyruvate |
US11116726B2 (en) | 2015-11-10 | 2021-09-14 | Childrens Research Institute, Childrens National Medical Center | Echinomycin formulation, method of making and method of use thereof |
SG11201803213XA (en) | 2015-11-12 | 2018-05-30 | Siamab Therapeutics Inc | Glycan-interacting compounds and methods of use |
CN108367079B (en) | 2015-11-12 | 2022-11-22 | 灰色视觉公司 | Aggregated microparticles for therapy |
TWI812873B (en) | 2015-11-30 | 2023-08-21 | 美商輝瑞股份有限公司 | Antibodies and antibody fragments for site-specific conjugation |
TWI726942B (en) | 2015-11-30 | 2021-05-11 | 美商輝瑞股份有限公司 | Site specific her2 antibody drug conjugates |
EP3383908A1 (en) | 2015-12-02 | 2018-10-10 | Stsciences, Inc. | Antibodies specific to glycosylated btla (b- and t- lymphocyte attenuator) |
KR20180086246A (en) | 2015-12-02 | 2018-07-30 | 주식회사 에스티큐브앤컴퍼니 | Antibodies and molecules that immunospecifically bind to BTN1A1 and their therapeutic uses |
US20190307857A1 (en) | 2015-12-09 | 2019-10-10 | Modernatx, Inc. | MODIFIED mRNA ENCODING A URIDINE DIPHOPSPHATE GLUCURONOSYL TRANSFERASE AND USES THEREOF |
US10548844B2 (en) | 2015-12-14 | 2020-02-04 | Massachusetts Institute Of Technology | pH-responsive mucoadhesive polymeric encapsulated microorganisms |
WO2017106630A1 (en) | 2015-12-18 | 2017-06-22 | The General Hospital Corporation | Polyacetal polymers, conjugates, particles and uses thereof |
JPWO2017110772A1 (en) | 2015-12-21 | 2018-09-13 | 富士フイルム株式会社 | Liposomes and liposome compositions |
EP4310503A3 (en) | 2015-12-30 | 2024-03-20 | Momenta Pharmaceuticals, Inc. | Methods related to biologics |
CN115814077A (en) | 2016-01-08 | 2023-03-21 | 供石公司 | Anti-pro/latent myostatin antibodies and methods of use thereof |
EP3405579A1 (en) | 2016-01-22 | 2018-11-28 | Modernatx, Inc. | Messenger ribonucleic acids for the production of intracellular binding polypeptides and methods of use thereof |
JOP20170017B1 (en) | 2016-01-25 | 2021-08-17 | Amgen Res Munich Gmbh | Pharmaceutical composition comprising bispecific antibody constructs |
LT3411402T (en) | 2016-02-03 | 2022-01-25 | Amgen Research (Munich) Gmbh | Bcma and cd3 bispecific t cell engaging antibody constructs |
EA201891753A1 (en) | 2016-02-03 | 2019-01-31 | Эмджен Рисерч (Мюник) Гмбх | BISPECIFIC CONSTRUCTIONS OF ANTIBODIES TO PSMA AND CD3, INVOLVING T-CELLS |
EA039859B1 (en) | 2016-02-03 | 2022-03-21 | Эмджен Рисерч (Мюник) Гмбх | Bispecific antibody constructs binding egfrviii and cd3 |
CN109071625A (en) | 2016-02-04 | 2018-12-21 | 柯瑞斯公司 | Smooth mutant and its application method |
US11357849B2 (en) | 2016-03-07 | 2022-06-14 | Musc Foundation For Research Development | Anti-nucleolin antibodies |
AU2017229232B2 (en) * | 2016-03-07 | 2022-12-15 | Memorial Sloan Kettering Cancer Center | Bone marrow-, reticuloendothelial system-, and/or lymph node-targeted radiolabeled liposomes and methods of their diagnostic and therapeutic use |
AU2017230103A1 (en) | 2016-03-11 | 2018-09-06 | Scholar Rock, Inc. | TGFβ1-binding immunoglobulins and use thereof |
WO2017157305A1 (en) | 2016-03-15 | 2017-09-21 | Generon (Shanghai) Corporation Ltd. | Multispecific fab fusion proteins and use thereof |
EP3445405A4 (en) | 2016-04-18 | 2019-12-18 | Sarepta Therapeutics, Inc. | Antisense oligomers and methods of using the same for treating diseases associated with the acid alpha-glucosidase gene |
WO2017181379A1 (en) | 2016-04-21 | 2017-10-26 | Versitech Limited | Compositions and methods for lightening skin and reducing hyperpigmentation |
CA3026880A1 (en) | 2016-06-08 | 2017-12-14 | Paul Foster | Treatment of igg4-related diseases with anti-cd19 antibodies crossbinding to cd32b |
ITUA20164630A1 (en) | 2016-06-23 | 2017-12-23 | Paolo Blasi | PHARMACOLOGICAL ADIUVANTS FOR TUMOR THERMAL WELDING |
WO2018004517A1 (en) | 2016-06-27 | 2018-01-04 | Alexion Pharmaceuticals, Inc. | Methods for treating hypophosphatasia in children and adolescents |
WO2018005657A1 (en) | 2016-06-28 | 2018-01-04 | Verily Life Sciences Llc | Serial filtration to generate small cholesterol-containing liposomes |
IL296127A (en) | 2016-07-22 | 2022-11-01 | Dana Farber Cancer Inst Inc | Glucocorticoid-induced tumor necrosis factor receptor (gitr) antibodies and methods of use thereof |
WO2018026890A1 (en) | 2016-08-03 | 2018-02-08 | Cymabay Therapeutics | Oxymethylene aryl compounds for treating inflammatory gastrointestinal diseases or gastrointestinal conditions |
TWI754659B (en) | 2016-08-08 | 2022-02-11 | 台灣微脂體股份有限公司 | Delivery vehicle, method and kit for preparing a liposomal composition containing mild acidic agent |
CA3035081A1 (en) | 2016-09-02 | 2018-03-08 | Dana-Farber Cancer Institute, Inc. | Composition and methods of treating b cell disorders |
MX2019002696A (en) | 2016-09-06 | 2019-09-27 | Dana Farber Cancer Inst Inc | Methods of treating or preventing zika virus infection. |
EP3509616A1 (en) | 2016-09-09 | 2019-07-17 | H. Hoffnabb-La Roche Ag | Selective peptide inhibitors of frizzled |
WO2018049248A1 (en) | 2016-09-09 | 2018-03-15 | Icellhealth Consulting Llc | Oncolytic virus equipped with bispecific engager molecules |
WO2018053180A2 (en) | 2016-09-14 | 2018-03-22 | The Trustees Of The University Of Pennsylvania | Proximity-based sortase-mediated protein purification and ligation |
CN109923126B (en) | 2016-09-16 | 2022-06-03 | 上海复宏汉霖生物技术股份有限公司 | anti-PD-1 antibodies |
EP3515488A1 (en) | 2016-09-23 | 2019-07-31 | Teva Pharmaceuticals International GmbH | Treating cluster headache |
PE20191148A1 (en) | 2016-09-23 | 2019-09-02 | Teva Pharmaceuticals Int Gmbh | TREATMENT OF REFRACTORY MIGRANA |
SG11201903674YA (en) | 2016-10-26 | 2019-05-30 | Modernatx Inc | Messenger ribonucleic acids for enhancing immune responses and methods of use thereof |
EA201990979A1 (en) | 2016-11-02 | 2019-09-30 | Ипсен Биофарм Лтд. | METHODS FOR TREATING GASTROINTESTINAL CANCER USING COMBINATION TYPES OF THERAPY CONTAINING LIPOSOMAL IRINOTECAN AND OXALALPLATIN |
US11779604B2 (en) | 2016-11-03 | 2023-10-10 | Kymab Limited | Antibodies, combinations comprising antibodies, biomarkers, uses and methods |
WO2018083535A1 (en) | 2016-11-04 | 2018-05-11 | Novimmune Sa | Anti-cd19 antibodies and methods of use thereof |
WO2018087720A1 (en) | 2016-11-14 | 2018-05-17 | Novartis Ag | Compositions, methods, and therapeutic uses related to fusogenic protein minion |
EP3541847A4 (en) | 2016-11-17 | 2020-07-08 | Seattle Genetics, Inc. | Glycan-interacting compounds and methods of use |
WO2018106738A1 (en) | 2016-12-05 | 2018-06-14 | Massachusetts Institute Of Technology | Brush-arm star polymers, conjugates and particles, and uses thereof |
PL3555064T3 (en) | 2016-12-16 | 2023-03-06 | Pfizer Inc. | Glp-1 receptor agonists and uses thereof |
US10772926B2 (en) | 2016-12-16 | 2020-09-15 | Nutragen Health Innovations, Inc. | Natural drugs for the treatment of inflammation and melanoma |
CR20190350A (en) | 2017-01-06 | 2019-11-15 | Scholar Rock Inc | ISOFORM-SPECIFIC, CONTEXT-PERMISSIVE TGFß1 INHIBITORS AND USE THEREOF |
ES2944357T3 (en) | 2017-01-06 | 2023-06-20 | Scholar Rock Inc | Treatment of metabolic diseases by inhibiting myostatin activation |
WO2018129395A1 (en) | 2017-01-06 | 2018-07-12 | Scholar Rock, Inc. | Methods for treating metabolic diseases by inhibiting myostatin activation |
WO2018144775A1 (en) | 2017-02-01 | 2018-08-09 | Modernatx, Inc. | Immunomodulatory therapeutic mrna compositions encoding activating oncogene mutation peptides |
JOP20190189A1 (en) | 2017-02-02 | 2019-08-01 | Amgen Res Munich Gmbh | Low ph pharmaceutical composition comprising t cell engaging antibody constructs |
WO2018152496A1 (en) | 2017-02-17 | 2018-08-23 | The Usa, As Represented By The Secretary, Dept. Of Health And Human Services | Compositions and methods for the diagnosis and treatment of zika virus infection |
PE20191487A1 (en) | 2017-03-03 | 2019-10-18 | Rinat Neuroscience Corp | ANTI-GITR ANTIBODIES AND METHODS OF USE OF THEM |
MA47812A (en) | 2017-03-03 | 2021-04-14 | Seagen Inc | COMPOUNDS INTERACTING WITH GLYCAN AND METHODS OF USE |
CA3055574A1 (en) | 2017-03-09 | 2018-09-13 | Cytomx Therapeutics, Inc. | Cd147 antibodies, activatable cd147 antibodies, and methods of making and use thereof |
WO2018183173A1 (en) | 2017-03-27 | 2018-10-04 | Boehringer Ingelheim International Gmbh | Anti il-36r antibodies combination therapy |
WO2018191153A1 (en) | 2017-04-09 | 2018-10-18 | The Cleveland Clinic Foundation | Cancer treatment by malat1 inhibition |
CA3059542A1 (en) | 2017-04-12 | 2018-10-18 | Pfizer Inc. | Antibodies having conditional affinity and methods of use thereof |
CA3059938A1 (en) | 2017-04-14 | 2018-10-18 | Kodiak Sciences Inc. | Complement factor d antagonist antibodies and conjugates thereof |
TW201902927A (en) | 2017-04-21 | 2019-01-16 | 美商梅利特公司 | Methods and antibodies for diabetes related applications |
EP3615569A1 (en) | 2017-04-25 | 2020-03-04 | The U.S.A. As Represented By The Secretary, Department Of Health And Human Services | Antibodies and methods for the diagnosis and treatment of epstein barr virus infection |
US11318190B2 (en) | 2017-05-05 | 2022-05-03 | United States Government As Represented By The Department Of Veterans Affairs | Methods and compositions for treating liver disease |
UY37726A (en) | 2017-05-05 | 2018-11-30 | Amgen Inc | PHARMACEUTICAL COMPOSITION THAT INCLUDES BISPECTIFIC ANTIBODY CONSTRUCTIONS FOR IMPROVED STORAGE AND ADMINISTRATION |
RU2019139817A (en) | 2017-05-10 | 2021-06-10 | Грейбуг Вижн, Инк. | DELAYED RELEASE MICROPARTICLES AND THEIR SUSPENSIONS FOR DRUG THERAPY |
US10994025B2 (en) | 2017-05-12 | 2021-05-04 | Massachusetts Institute Of Technology | Argonaute protein-double stranded RNA complexes and uses related thereto |
SG11201909751TA (en) | 2017-05-12 | 2019-11-28 | Augusta University Research Institute Inc | Human alpha fetoprotein-specific t cell receptors and uses thereof |
JP7220161B2 (en) | 2017-05-26 | 2023-02-09 | ノビミューン エスアー | Anti-CD47x anti-mesothelin antibodies and methods of using same |
CA3065301A1 (en) | 2017-05-31 | 2018-12-06 | Stcube & Co., Inc. | Antibodies and molecules that immunospecifically bind to btn1a1 and the therapeutic uses thereof |
KR20200024158A (en) | 2017-05-31 | 2020-03-06 | 주식회사 에스티큐브앤컴퍼니 | How to treat cancer using antibodies and molecules that immunospecifically bind to BTN1A1 |
AU2018278327B2 (en) | 2017-06-01 | 2023-03-16 | Cytomx Therapeutics, Inc. | Activatable anti-pdl1 antibodies and methods of use thereof |
JP2020522562A (en) | 2017-06-06 | 2020-07-30 | ストキューブ アンド シーオー., インコーポレイテッド | Methods of treating cancer with antibodies and molecules that bind to BTN1A1 or BTN1A1 ligand |
GB201710838D0 (en) | 2017-07-05 | 2017-08-16 | Ucl Business Plc | Bispecific antibodies |
CN110832077A (en) | 2017-07-06 | 2020-02-21 | 箭头药业股份有限公司 | RNAi agents for inhibiting α -ENaC expression and methods of use |
CA3069179A1 (en) | 2017-07-13 | 2019-01-17 | Massachusetts Institute Of Technology | Targeting the hdac2-sp3 complex to enhance synaptic function |
KR20200027971A (en) | 2017-07-14 | 2020-03-13 | 싸이톰스 테라퓨틱스, 인크. | Anti-CD166 antibodies and uses thereof |
WO2019018629A1 (en) | 2017-07-19 | 2019-01-24 | The Usa, As Represented By The Secretary, Dept. Of Health And Human Services | Antibodies and methods for the diagnosis and treatment of hepatitis b virus infection |
JP2020530554A (en) | 2017-07-20 | 2020-10-22 | シートムエックス セラピューティクス,インコーポレイテッド | Methods and Uses for Qualitative and / or Quantitative Analysis of Activating Antibody Properties |
MA49634A (en) | 2017-07-21 | 2020-05-27 | Modernatx Inc | MODIFIED RNA CODING FOR A PROPIONYL-COA-CARBOXYLASE AND ASSOCIATED USES |
WO2019016784A1 (en) | 2017-07-21 | 2019-01-24 | Universidade De Coimbra | Anti-nucleolin antibody |
MA49684A (en) | 2017-07-24 | 2020-06-03 | Modernatx Inc | GLUCOSE-6-PHOSPHATASE MODIFIED RNA AND ASSOCIATED USES |
PT3658184T (en) | 2017-07-27 | 2023-11-29 | Alexion Pharma Inc | High concentration anti-c5 antibody formulations |
EP3658583A1 (en) | 2017-07-28 | 2020-06-03 | Scholar Rock, Inc. | Ltbp complex-specific inhibitors of tgf-beta 1 and uses thereof |
BR112020002280A2 (en) | 2017-08-03 | 2020-07-28 | Otsuka Pharmaceutical Co., Ltd. | pharmacological compound and purification methods |
US11135187B2 (en) | 2017-08-22 | 2021-10-05 | National Institutes Of Health (Nih) | Compositions and methods for treating diabetic retinopathy |
KR20200058406A (en) | 2017-08-30 | 2020-05-27 | 싸이톰스 테라퓨틱스, 인크. | Activatable anti-CD166 antibodies and methods of use |
EP3679070A1 (en) | 2017-09-07 | 2020-07-15 | Augusta University Research Institute, Inc. | Antibodies to programmed cell death protein 1 |
US11364303B2 (en) | 2017-09-29 | 2022-06-21 | Pfizer Inc. | Cysteine engineered antibody drug conjugates |
EP3694890A4 (en) | 2017-10-12 | 2021-11-03 | Immunowake Inc. | Vegfr-antibody light chain fusion protein |
KR20200064096A (en) | 2017-10-14 | 2020-06-05 | 싸이톰스 테라퓨틱스, 인크. | Antibodies, activatable antibodies, bispecific antibodies and bispecific activatable antibodies and methods of using them |
WO2019077496A1 (en) | 2017-10-17 | 2019-04-25 | Rhizen Pharmaceuticals Sa | Crac channel modulators for treating esophageal cancer |
US11555189B2 (en) | 2017-10-18 | 2023-01-17 | Sarepta Therapeutics, Inc. | Antisense oligomer compounds |
WO2019082124A1 (en) | 2017-10-26 | 2019-05-02 | Rhizen Pharmaceuticals Sa | Composition and method for treating diffuse large b-cell lymphoma |
BR112020008127A2 (en) | 2017-10-27 | 2020-10-13 | Pfizer Inc. | antibodies and antibody-drug conjugates specific for cd123 and their uses |
JP2021501160A (en) | 2017-10-30 | 2021-01-14 | ルヒゼン ファーマスティカルズ エスエー | Calcium Release Activated Calcium Channel Modulator for the Treatment of Hematological and Solid Cancers |
WO2019086626A1 (en) | 2017-11-03 | 2019-05-09 | Centro Nacional De Investigaciones Cardiovasculares Carlos Iii (F.S.P.) | MIRNAs AND COMBINATIONS THEREOF FOR USE IN THE TREATMENT OF HUMAN B CELL NEOPLASIAS |
EP3720557A1 (en) | 2017-12-06 | 2020-10-14 | Rhizen Pharmaceuticals S.A. | Composition and method for treating peripheral t-cell lymphoma and cutaneous t-cell lymphoma |
US11946094B2 (en) | 2017-12-10 | 2024-04-02 | Augusta University Research Institute, Inc. | Combination therapies and methods of use thereof |
BR112020011627A2 (en) | 2017-12-11 | 2020-11-17 | Amgen Inc. | continuous manufacturing process for bispecific antibody products |
TW201940518A (en) | 2017-12-29 | 2019-10-16 | 美商安進公司 | Bispecific antibody construct directed to MUC17 and CD3 |
WO2019165101A1 (en) | 2018-02-22 | 2019-08-29 | Verily Life Sciences Llc | Combining orthogonal chemistries for preparation of multiplexed nanoparticles |
MX2020008455A (en) | 2018-02-28 | 2021-10-26 | Pfizer | Il-15 variants and uses thereof. |
WO2019173771A1 (en) | 2018-03-09 | 2019-09-12 | Cytomx Therapeutics, Inc. | Activatable cd147 antibodies and methods of making and use thereof |
AU2019235523A1 (en) | 2018-03-14 | 2020-10-29 | Novimmune Sa | Anti-CD3 epsilon antibodies and methods of use thereof |
US11485782B2 (en) | 2018-03-14 | 2022-11-01 | Beijing Xuanyi Pharmasciences Co., Ltd. | Anti-claudin 18.2 antibodies |
WO2019183320A1 (en) | 2018-03-21 | 2019-09-26 | Colorado State University Research Foundation | Cancer vaccine compositions and methods of use thereof |
US10722528B2 (en) | 2018-03-28 | 2020-07-28 | Augusta University Research Institute, Inc. | Compositions and methods for inhibiting metastasis |
JP2021520781A (en) | 2018-04-06 | 2021-08-26 | チルドレンズ メディカル センター コーポレーションChildren’S Medical Center Corporation | Compositions and Methods for Somatic Cell Reprogramming and Imprinting Modulation |
CN112334485A (en) | 2018-04-06 | 2021-02-05 | 百进生物科技公司 | Anti-tetraspanin 33agents and compositions thereof and methods of making and using |
US20210115156A1 (en) | 2018-04-13 | 2021-04-22 | INSERM (Institut National de la Santé et de la Recherche Médicale) | Fc-engineered anti-human ige antibodies and methods of use |
EP3788071A1 (en) | 2018-05-02 | 2021-03-10 | The United States Of America, As Represented By The Secretary, Department of Health and Human Services | Antibodies and methods for the diagnosis, prevention, and treatment of epstein barr virus infection |
SG11202010934SA (en) | 2018-05-23 | 2020-12-30 | Pfizer | Antibodies specific for gucy2c and uses thereof |
KR102602329B1 (en) | 2018-05-23 | 2023-11-16 | 화이자 인코포레이티드 | Antibodies specific for CD3 and their uses |
US20210196823A1 (en) | 2018-06-08 | 2021-07-01 | Pfizer Inc. | Methods of Treating Metabolic Disease |
CA3045644C (en) | 2018-06-13 | 2024-01-16 | Pfizer Inc. | Glp-1 receptor agonists and uses thereof |
CN112566637B (en) | 2018-06-15 | 2023-11-14 | 辉瑞公司 | GLP-1 receptor agonists and uses thereof |
US20210347842A1 (en) | 2018-06-19 | 2021-11-11 | Eli Lilly And Company | Compositions and methods of use of il-10 agents in conjunction with chimeric antigen receptor cell therapy |
BR112020025824A2 (en) | 2018-06-28 | 2021-03-23 | Crispr Therapeutics Ag | compositions and methods for genome editing by inserting donor polynucleotides |
KR20210027436A (en) | 2018-06-29 | 2021-03-10 | 베링거 인겔하임 인터내셔날 게엠베하 | Anti-CD40 antibodies for use in treating autoimmune diseases |
TWI709562B (en) | 2018-07-06 | 2020-11-11 | 美商輝瑞股份有限公司 | Manufacturing process and intermediates for a pyrrolo〔2,3-d〕pyrimidine compound and use thereof |
US20210340238A1 (en) | 2018-07-11 | 2021-11-04 | Scholar Rock, Inc. | TGFß1 INHIBITORS AND USE THEREOF |
EP3677278B1 (en) | 2018-07-11 | 2021-11-10 | Scholar Rock, Inc. | Isoform selective tgfbeta1 inhibitors and use thereof |
EP3820508A1 (en) | 2018-07-11 | 2021-05-19 | Scholar Rock, Inc. | High-affinity, isoform-selective tgf?1 inhibitors and use thereof |
AU2019306165A1 (en) | 2018-07-20 | 2021-02-25 | Pierre Fabre Medicament | Receptor for vista |
EP3837367A1 (en) | 2018-08-16 | 2021-06-23 | Alnylam Pharmaceuticals, Inc. | Compositions and methods for inhibiting expression of the lect2 gene |
EP3849515A1 (en) * | 2018-09-11 | 2021-07-21 | Memorial Sloan Kettering Cancer Center | Bone marrow-, reticuloendothelial system-, and/or lymph node-targeted radiolabeled liposomes and methods of their diagnostic and therapeutic use |
CN113365697A (en) | 2018-09-25 | 2021-09-07 | 百进生物科技公司 | anti-TLR9 agents and compositions and methods of making and using the same |
SG11202103025PA (en) | 2018-09-27 | 2021-04-29 | Phosphogam Inc | Methods and compositions for the expansion and use of allogeneic gamma/delta-t cells |
CA3114567A1 (en) | 2018-09-28 | 2020-04-02 | Lyvgen Biopharma Co., Ltd. | Anti-cd40 binding molecules having engineered fc domains and therapeutic uses thereof |
KR20210074341A (en) | 2018-10-10 | 2021-06-21 | 베링거 인겔하임 인터내셔날 게엠베하 | Methods for membrane gas transfer in high-density bioreactor cultures |
MX2021003976A (en) | 2018-10-11 | 2021-05-27 | Amgen Inc | Downstream processing of bispecific antibody constructs. |
WO2020079652A1 (en) | 2018-10-17 | 2020-04-23 | Insilico Medicine Hong Kong Limited | Kinase inhibitors |
BR112021007765A2 (en) | 2018-10-23 | 2021-08-03 | Scholar Rock, Inc. | selective rgmc inhibitors and their use |
CN112955252B (en) * | 2018-10-26 | 2023-11-21 | 康涅狄格大学 | Continuous processing system and method for internal and external modification of nanoparticles |
US20220023439A1 (en) | 2018-11-02 | 2022-01-27 | Cytomx Therapeutics, Inc. | Activatable anti-cd166 antibodies and methods of use thereof |
KR20210106447A (en) | 2018-11-22 | 2021-08-30 | 치루 레고르 테라퓨틱스 인코포레이티드 | GLP-1R agonists and uses thereof |
WO2020113135A1 (en) | 2018-11-29 | 2020-06-04 | Flagship Pioneering Innovations V, Inc. | Methods of modulating rna |
CN113271956A (en) | 2018-12-06 | 2021-08-17 | 西托姆克斯治疗公司 | Matrix metalloprotease cleavable and serine or cysteine protease cleavable substrates and methods of use thereof |
CA3177829A1 (en) | 2018-12-12 | 2020-06-18 | Kite Pharma, Inc. | Chimeric antigen and t cell receptors and methods of use |
KR20210131335A (en) | 2019-01-22 | 2021-11-02 | 메사추세츠 인스티튜트 오브 테크놀로지 | Human blood brain barrier in vitro |
JP2020117502A (en) | 2019-01-28 | 2020-08-06 | ファイザー・インク | Method of treating signs and symptoms of osteoarthritis |
WO2020160291A2 (en) | 2019-01-30 | 2020-08-06 | Scholar Rock, Inc. | LTBP COMPLEX-SPECIFIC INHIBITORS OF TGFβ AND USES THEREOF |
CA3130034C (en) | 2019-02-15 | 2023-12-19 | Pfizer Inc. | Crystalline pyrimidinyl-3,8-diazabicyclo[3.2.1]octanylmethanone compound and use thereof |
WO2020170103A1 (en) | 2019-02-18 | 2020-08-27 | Pfizer Inc. | Method of treatment of chronic low back pain |
US20220275059A1 (en) | 2019-02-20 | 2022-09-01 | Harbour Antibodies Bv | Antibodies |
US11739078B2 (en) | 2019-02-22 | 2023-08-29 | Insilico Medicine Ip Limited | Methods of inhibiting kinases |
WO2020176672A1 (en) | 2019-02-26 | 2020-09-03 | Cytomx Therapeutics, Inc. | Combined therapies of activatable immune checkpoint inhibitors and conjugated activatable antibodies |
US20200283796A1 (en) | 2019-03-05 | 2020-09-10 | Massachusetts Institute Of Technology | Dna launched rna replicon system (drep) and uses thereof |
WO2020185293A1 (en) | 2019-03-08 | 2020-09-17 | Massachusetts Institute Of Technology | Synthetic oncolytic lnp-replicon rna and uses for cancer immunotherapy |
SG11202110626SA (en) | 2019-03-27 | 2021-10-28 | Umc Utrecht Holding Bv | Engineered iga antibodies and methods of use |
CA3133652A1 (en) | 2019-04-01 | 2020-10-08 | Genentech, Inc. | Compositions and methods for stabilizing protein-containing formulations |
WO2020201991A1 (en) | 2019-04-02 | 2020-10-08 | Array Biopharma Inc. | Protein tyrosine phosphatase inhibitors |
TW202115086A (en) | 2019-06-28 | 2021-04-16 | 美商輝瑞大藥廠 | Bckdk inhibitors |
CA3144848C (en) | 2019-06-28 | 2023-11-21 | Pfizer Inc. | 5-(thiophen-2-yl)-1h-tetrazole derivatives as bckdk inhibitors useful for treating various diseases |
LT6699B (en) | 2019-07-15 | 2020-02-10 | UAB "Valentis" | Method for production of proliposomes by using up to 5% ethanol and the use thereof for encapsulation of lipophilic substances |
CN112300279A (en) | 2019-07-26 | 2021-02-02 | 上海复宏汉霖生物技术股份有限公司 | Methods and compositions directed to anti-CD 73 antibodies and variants |
US10758329B1 (en) | 2019-08-20 | 2020-09-01 | Raymond L. Wright, III | Hydrating mouth guard |
JP2022546570A (en) | 2019-09-03 | 2022-11-04 | アルナイラム ファーマシューティカルズ, インコーポレイテッド | Compositions and methods for inhibiting expression of the LECT2 gene |
CN114650844A (en) | 2019-09-23 | 2022-06-21 | 西托姆克斯治疗公司 | anti-CD 47 antibodies, activatable anti-CD 47 antibodies, and methods of use thereof |
WO2021062323A1 (en) | 2019-09-26 | 2021-04-01 | Stcube & Co. | Antibodies specific to glycosylated ctla-4 and methods of use thereof |
US11492622B2 (en) | 2019-09-26 | 2022-11-08 | Massachusetts Institute Of Technology | MicroRNA-based logic gates and uses thereof |
US11834659B2 (en) | 2019-09-26 | 2023-12-05 | Massachusetts Institute Of Technology | Trans-activated functional RNA by strand displacement and uses thereof |
WO2021067526A1 (en) | 2019-10-02 | 2021-04-08 | Alexion Pharmaceuticals, Inc. | Complement inhibitors for treating drug-induced complement-mediated response |
TWI771766B (en) | 2019-10-04 | 2022-07-21 | 美商輝瑞股份有限公司 | Diacylglycerol acyltransferase 2 inhibitor |
EP4038189A1 (en) | 2019-10-04 | 2022-08-10 | Alnylam Pharmaceuticals, Inc. | Compositions and methods for silencing ugt1a1 gene expression |
WO2021071830A1 (en) | 2019-10-07 | 2021-04-15 | University Of Virginia Patent Foundation | Modulating lymphatic vessels in neurological disease |
CN114829404A (en) | 2019-10-09 | 2022-07-29 | 斯特库比公司 | Antibodies specific for glycosylated LAG3 and methods of use thereof |
AU2020364071A1 (en) | 2019-10-10 | 2022-05-26 | Kodiak Sciences Inc. | Methods of treating an eye disorder |
EP4041773A1 (en) | 2019-10-11 | 2022-08-17 | Beth Israel Deaconess Medical Center, Inc. | Anti-tn antibodies and uses thereof |
EP4048251A1 (en) | 2019-10-21 | 2022-08-31 | Rhizen Pharmaceuticals AG | Compositions comprising a dhodh inhibitor for the treatment of acute myeloid leukemia |
US20230040920A1 (en) | 2019-11-01 | 2023-02-09 | Alnylam Pharmaceuticals, Inc. | Compositions and methods for silencing dnajb1-prkaca fusion gene expression |
CN115175893A (en) | 2019-12-10 | 2022-10-11 | 辉瑞公司 | Solid forms of 2- ((4- ((S) -2- (5-chloropyridin-2-yl) -2-methylbenzo [ d ] [1,3] dioxol-4-yl) piperidin-1-yl) methyl) -1- (((S) -oxetan-2-yl) methyl) -1H-benzo [ d ] imidazole-6-carboxylic acid, 1, 3-dihydroxy-2- (hydroxymethyl) propan-2-amine salt |
WO2021142427A1 (en) | 2020-01-11 | 2021-07-15 | Scholar Rock, Inc. | TGFβ INHIBITORS AND USE THEREOF |
US20230050148A1 (en) | 2020-01-11 | 2023-02-16 | Scholar Rock, Inc. | Tgf-beta inhibitors and use thereof |
CN115315273A (en) | 2020-01-14 | 2022-11-08 | 辛德凯因股份有限公司 | IL-2 orthologs and methods of use thereof |
CN115427447A (en) | 2020-01-17 | 2022-12-02 | 百进生物科技公司 | anti-TLR 7 agents and compositions and methods of making and using the same |
US20230067811A1 (en) | 2020-01-24 | 2023-03-02 | University Of Virginia Patent Foundation | Modulating lymphatic vessels in neurological disease |
WO2021163066A1 (en) | 2020-02-10 | 2021-08-19 | Alnylam Pharmaceuticals, Inc. | Compositions and methods for silencing vegf-a expression |
JP2022058085A (en) | 2020-02-24 | 2022-04-11 | ファイザー・インク | Combination of inhibitors of diacylglycerol acyltransferase 2 and inhibitors of acetyl-coa carboxylase |
CA3170563A1 (en) | 2020-03-06 | 2021-09-10 | Raymond P. Goodrich | Production of vaccines comprising inactivated sars-cov-2 viral particles |
CA3174908A1 (en) | 2020-03-09 | 2021-09-16 | Pfizer Inc. | Fusion proteins and uses thereof |
GB202003632D0 (en) | 2020-03-12 | 2020-04-29 | Harbour Antibodies Bv | SARS-Cov-2 (SARS2, COVID-19) antibodies |
WO2021188096A1 (en) | 2020-03-17 | 2021-09-23 | Dst Pharma, Inc. | Methods and compositions for treating viral infections |
CN115666523A (en) | 2020-03-26 | 2023-01-31 | Plx 奥普科有限公司 | Drug carriers capable of pH dependent reconstitution and methods of making and using the same |
WO2021191812A1 (en) | 2020-03-27 | 2021-09-30 | Pfizer Inc. | Treatment of type 2 diabetes or obesity or overweight with 2-[(4-{6-[(4-cyano-2-fluorobenzyl)oxy]pyridin-2-yl} piperidin-1-yl)methyl]-1-[(2s)-oxetan-2-ylmethyl]-1h-benzimidazole-6-carboxylic acid or a pharmaceutically salt thereof |
WO2021202443A2 (en) | 2020-03-30 | 2021-10-07 | Alnylam Pharmaceucticals, Inc. | Compositions and methods for silencing dnajc15 gene expression |
BR112022020145A2 (en) | 2020-04-06 | 2023-01-03 | Alnylam Pharmaceuticals Inc | COMPOSITIONS AND METHODS FOR SILENCING THE MYOC EXPRESSION |
TW202204617A (en) | 2020-04-07 | 2022-02-01 | 美商艾爾妮蘭製藥公司 | Compositions and methods for silencing scn9a expression |
CA3179566A1 (en) | 2020-04-08 | 2021-10-14 | Pfizer Inc. | Crystalline forms of 3-cyano-1-[4-[6-(1-methyl-1h-pyrazol-4-yl)pyrazolo[1,5-a]pyrazin-4-yl]-1h-pyrazol-1-yl]cyclobutaneacetonitrile, and use thereof |
BR112022021450A2 (en) | 2020-04-24 | 2022-12-27 | Millennium Pharm Inc | CD19 OR LEADING FRAGMENT, METHOD OF TREATMENT OF A CANCER, PHARMACEUTICAL COMPOSITION, NUCLEIC ACID, VECTOR AND, ISOLATED CELL |
US20230181750A1 (en) | 2020-05-06 | 2023-06-15 | Crispr Therapeutics Ag | Mask peptides and masked anti-ptk7 antibodies comprising such |
US20210355463A1 (en) | 2020-05-15 | 2021-11-18 | Crispr Therapeutics Ag | Messenger rna encoding cas9 for use in genome-editing systems |
WO2021237097A1 (en) | 2020-05-21 | 2021-11-25 | Alnylam Pharmaceuticals, Inc. | Compositions and methods for inhibiting marc1 gene expression |
EP4157832A1 (en) | 2020-05-27 | 2023-04-05 | Qilu Regor Therapeutics Inc. | Salt and crystal forms of glp-1r agonists and uses thereof |
MX2022015706A (en) | 2020-06-09 | 2023-01-24 | Pfizer | Spiro compounds as melanocortin 4 receptor antagonists and uses thereof. |
BR112022025667A2 (en) | 2020-06-26 | 2023-03-07 | Pfizer | METHODS OF TREATMENT OF INFLAMMATORY BOWEL DISEASE WITH TL1A ANTIBODIES |
WO2022006562A1 (en) | 2020-07-03 | 2022-01-06 | Dana-Farber Cancer Institute, Inc. | Multispecific coronavirus antibodies |
BR112023000701A2 (en) | 2020-07-17 | 2023-02-07 | Pfizer | THERAPEUTIC ANTIBODIES AND THEIR USES |
CN116710079A (en) | 2020-07-24 | 2023-09-05 | 斯特兰德生物科技公司 | Lipid nanoparticles comprising modified nucleotides |
US20220041595A1 (en) | 2020-07-28 | 2022-02-10 | Jazz Pharmaceuticals Ireland Limited | Chiral synthesis of fused bicyclic raf inhibitors |
EP4188552A2 (en) | 2020-07-28 | 2023-06-07 | Jazz Pharmaceuticals Ireland Limited | Fused bicyclic raf inhibitors and methods for use thereof |
EP4192863A2 (en) | 2020-08-05 | 2023-06-14 | Synthekine, Inc. | Il2rg binding molecules and methods of use |
AU2021322238A1 (en) | 2020-08-05 | 2023-03-23 | Synthekine, Inc. | Compositions and methods related to IL27 receptor binding |
EP4192489A2 (en) | 2020-08-05 | 2023-06-14 | Synthekine, Inc. | Il2rb binding molecules and methods of use |
KR20230061394A (en) | 2020-08-05 | 2023-05-08 | 신테카인, 인크. | IL10Ra Binding Molecules and Methods of Use |
AR123156A1 (en) | 2020-08-06 | 2022-11-02 | Qilu Regor Therapeutics Inc | GLP-1R AGONISTS AND THEIR USES |
JP2023538012A (en) | 2020-08-14 | 2023-09-06 | カイト ファーマ インコーポレイテッド | Improving immune cell function |
US20230365675A1 (en) | 2020-09-14 | 2023-11-16 | Vor Biopharma Inc. | Single domain antibodies against cd33 |
US20220089759A1 (en) | 2020-09-21 | 2022-03-24 | Boehringer Ingelheim International Gmbh | Use of anti-cd40 antibodies for treatment of inflammatory conditions |
JP2023544200A (en) | 2020-10-06 | 2023-10-20 | ゼンコー・インコーポレイテッド | Biomarkers, methods, and compositions for treating autoimmune diseases including systemic lupus erythematosus (SLE) |
JP2023546125A (en) | 2020-10-14 | 2023-11-01 | キル・レガー・セラピューティクス・インコーポレーテッド | Crystal forms of GLP-1R agonists and their uses |
WO2022093641A1 (en) | 2020-10-30 | 2022-05-05 | BioLegend, Inc. | Anti-nkg2a agents and compositions and methods for making and using the same |
WO2022093640A1 (en) | 2020-10-30 | 2022-05-05 | BioLegend, Inc. | Anti-nkg2c agents and compositions and methods for making and using the same |
US11058637B1 (en) * | 2020-11-25 | 2021-07-13 | King Abdulaziz University | Surface-modified emulsomes for intranasal delivery of drugs |
CA3200234A1 (en) | 2020-11-25 | 2022-06-02 | Daryl C. Drummond | Lipid nanoparticles for delivery of nucleic acids, and related methods of use |
AU2021400221A1 (en) | 2020-12-18 | 2023-07-13 | Momenta Pharmaceuticals, Inc. | Antibodies against integrin alpha 11 beta 1 |
TW202241931A (en) | 2021-01-08 | 2022-11-01 | 美商斯特蘭德治療股份有限公司 | Expression constructs and uses thereof |
US11278494B1 (en) | 2021-01-22 | 2022-03-22 | Pacira Pharmaceuticals, Inc. | Manufacturing of bupivacaine multivesicular liposomes |
US11033495B1 (en) | 2021-01-22 | 2021-06-15 | Pacira Pharmaceuticals, Inc. | Manufacturing of bupivacaine multivesicular liposomes |
US11357727B1 (en) | 2021-01-22 | 2022-06-14 | Pacira Pharmaceuticals, Inc. | Manufacturing of bupivacaine multivesicular liposomes |
WO2022170008A2 (en) | 2021-02-05 | 2022-08-11 | Boehringer Ingelheim International Gmbh | Anti-il1rap antibodies |
JP2024508818A (en) | 2021-02-24 | 2024-02-28 | インシリコ メディシン アイピー リミテッド | Analogs for the treatment of diseases |
WO2022192439A1 (en) | 2021-03-11 | 2022-09-15 | Kite Pharma, Inc. | Improving immune cell function |
WO2022195504A1 (en) | 2021-03-19 | 2022-09-22 | Pfizer Inc. | Method of treating osteoarthritis pain with an anti ngf antibody |
IL305827A (en) | 2021-03-22 | 2023-11-01 | Novimmune Sa | Bispecific antibodies targeting cd47 and pd-l1 and methods of use thereof |
EP4313309A1 (en) | 2021-03-22 | 2024-02-07 | Novimmune S.A. | Bispecific antibodies targeting cd47 and pd-l1 and methods of use thereof |
WO2022204581A2 (en) | 2021-03-26 | 2022-09-29 | Scholar Rock, Inc. | Tgf-beta inhibitors and use thereof |
EP4320149A1 (en) | 2021-04-09 | 2024-02-14 | Takeda Pharmaceutical Company Limited | Antibodies targeting complement factor d and uses therof |
EP4326873A1 (en) | 2021-04-22 | 2024-02-28 | Dana-Farber Cancer Institute, Inc. | Compositions and methods for treating cancer |
US20230018888A1 (en) | 2021-04-26 | 2023-01-19 | Millennium Pharmaceuticals, Inc. | Anti-adgre2 antibodies and uses thereof |
WO2022232044A2 (en) | 2021-04-26 | 2022-11-03 | Millennium Pharmaceuticals, Inc. | Anti-clec12a antibodies and uses thereof |
WO2022235940A1 (en) | 2021-05-06 | 2022-11-10 | Dana-Farber Cancer Institute, Inc. | Antibodies against alk and methods of use thereof |
US20220372114A1 (en) | 2021-05-17 | 2022-11-24 | Curia Ip Holdings, Llc | Sars-cov-2 spike protein antibodies |
US20230115257A1 (en) | 2021-05-17 | 2023-04-13 | Curia Ip Holdings, Llc | Sars-cov-2 spike protein antibodies |
WO2022256723A2 (en) | 2021-06-03 | 2022-12-08 | Scholar Rock, Inc. | Tgf-beta inhibitors and therapeutic use thereof |
WO2022271867A1 (en) | 2021-06-23 | 2022-12-29 | Scholar Rock, Inc. | A myostatin pathway inhibitor in combination with a glp-1 pathway activator for use in treating metabolic disorders |
WO2023283403A2 (en) | 2021-07-09 | 2023-01-12 | Alnylam Pharmaceuticals, Inc. | Bis-rnai compounds for cns delivery |
TW202306985A (en) | 2021-07-12 | 2023-02-16 | 美商建南德克公司 | Structures for reducing antibody-lipase binding |
WO2023288277A1 (en) | 2021-07-14 | 2023-01-19 | Scholar Rock, Inc. | Ltbp complex-specific inhibitors of tgfb1 and uses thereof |
WO2023007374A1 (en) | 2021-07-27 | 2023-02-02 | Pfizer Inc. | Method of treatment of cancer pain with tanezumab |
GB202111758D0 (en) | 2021-08-17 | 2021-09-29 | Cantab Biopharmaceuticals Patents Ltd | Modified colloidal particles for use in the treatment of haemophilia A |
GB202111759D0 (en) | 2021-08-17 | 2021-09-29 | Cantab Biopharmaceuticals Patents Ltd | Modified colloidal particles |
GB202111757D0 (en) | 2021-08-17 | 2021-09-29 | Cantab Biopharmaceuticals Patents Ltd | Modified colloidal particles for use in the treatment of haemophilia A |
CA3230347A1 (en) | 2021-08-31 | 2023-03-09 | Pfizer Inc. | Solid forms of 2-[(4-{6-[(4-cyano-2-fluorobenzyl)oxy]pyridin-2-yl}piperidin-1-yl)methyl]-1-[(2s)-oxetan-2-ylmethyl]-1h-benzimidazole-6-carboxylic acid, 1,3-dihydroxy-2-(hydroxymethyl)propan-2-amine salt |
WO2023034901A1 (en) | 2021-09-01 | 2023-03-09 | The Broad Institute, Inc. | Tumor avatar vaccine compositions and uses thereof |
WO2023036982A1 (en) | 2021-09-10 | 2023-03-16 | Harbour Antibodies Bv | Anti-sars2-s antibodies |
GB202112935D0 (en) | 2021-09-10 | 2021-10-27 | Harbour Antibodies Bv | Sars-cov-2 (sars2, covid-19) heavy chain only antibodies |
TW202330612A (en) | 2021-10-20 | 2023-08-01 | 日商武田藥品工業股份有限公司 | Compositions targeting bcma and methods of use thereof |
WO2023077148A1 (en) | 2021-11-01 | 2023-05-04 | Tome Biosciences, Inc. | Single construct platform for simultaneous delivery of gene editing machinery and nucleic acid cargo |
WO2023081471A1 (en) | 2021-11-05 | 2023-05-11 | Dana-Farber Cancer Institute, Inc. | Human broadly crossreactive influenza monoclonal antibodies and methods of use thereof |
WO2023097024A1 (en) | 2021-11-24 | 2023-06-01 | Dana-Farber Cancer Institute, Inc. | Antibodies against ctla-4 and methods of use thereof |
WO2023100061A1 (en) | 2021-12-01 | 2023-06-08 | Pfizer Inc. | 3-phenyl-1-benzothiophene-2-carboxylic acid derivatives as branched-chain alpha keto acid dehydrogenase kinase inhibitors for the treatment of diabetes, kidney diseases, nash and heart failure |
WO2023105387A1 (en) | 2021-12-06 | 2023-06-15 | Pfizer Inc. | Melanocortin 4 receptor antagonists and uses thereof |
WO2023105371A1 (en) | 2021-12-08 | 2023-06-15 | Array Biopharma Inc. | Crystalline form of n-(2-chloro-3-((5-chloro-3-methyl-4-oxo-3,4-dihydroquinazolin-6-yl)amino)-4-fluorophenyl)-3-fluoroazetidine-1-sulfonamide |
WO2023114543A2 (en) | 2021-12-17 | 2023-06-22 | Dana-Farber Cancer Institute, Inc. | Platform for antibody discovery |
WO2023111817A1 (en) | 2021-12-17 | 2023-06-22 | Pfizer Inc. | Crystalline forms of [(1r,5s,6r)-3-{2-[(2s)-2-methylazetidin-1-yl]-6-(trifluoromethyl) pyrimidin-4-yl}-3-azabicyclo[3.1.0]hex-6-yl]acetic acid |
WO2023114544A1 (en) | 2021-12-17 | 2023-06-22 | Dana-Farber Cancer Institute, Inc. | Antibodies and uses thereof |
WO2023122764A1 (en) | 2021-12-22 | 2023-06-29 | Tome Biosciences, Inc. | Co-delivery of a gene editor construct and a donor template |
WO2023178357A1 (en) | 2022-03-18 | 2023-09-21 | Evolveimmune Therapeutics, Inc. | Bispecific antibody fusion molecules and methods of use thereof |
WO2023201238A1 (en) | 2022-04-11 | 2023-10-19 | Vor Biopharma Inc. | Binding agents and methods of use thereof |
WO2023205744A1 (en) | 2022-04-20 | 2023-10-26 | Tome Biosciences, Inc. | Programmable gene insertion compositions |
WO2023212618A1 (en) | 2022-04-26 | 2023-11-02 | Strand Therapeutics Inc. | Lipid nanoparticles comprising venezuelan equine encephalitis (vee) replicon and uses thereof |
WO2023215831A1 (en) | 2022-05-04 | 2023-11-09 | Tome Biosciences, Inc. | Guide rna compositions for programmable gene insertion |
WO2023220641A2 (en) | 2022-05-11 | 2023-11-16 | Juno Therapeutics, Inc. | Methods and uses related to t cell therapy and production of same |
WO2023225670A2 (en) | 2022-05-20 | 2023-11-23 | Tome Biosciences, Inc. | Ex vivo programmable gene insertion |
WO2023228023A1 (en) | 2022-05-23 | 2023-11-30 | Pfizer Inc. | Treatment of type 2 diabetes or weight management control with 2-((4-((s)-2-(5-chloropyridin-2-yl)-2-methylbenzo[d][1,3]dioxol-4-yl)piperidin-1-yl)methyl)-1-(((s)-oxetan-2-yl)methyl)-1h-benzo[d]imidazole-6-carboxylic acid or a pharmaceutically salt thereof |
WO2023235852A1 (en) | 2022-06-03 | 2023-12-07 | Zenas Biopharma, Inc. | Methods and compositions for treating igg4- related diseases |
WO2024020051A1 (en) | 2022-07-19 | 2024-01-25 | BioLegend, Inc. | Anti-cd157 antibodies, antigen-binding fragments thereof and compositions and methods for making and using the same |
WO2024030858A1 (en) | 2022-08-01 | 2024-02-08 | Cytomx Therapeutics, Inc. | Protease-cleavable substrates and methods of use thereof |
WO2024030850A1 (en) | 2022-08-01 | 2024-02-08 | Cytomx Therapeutics, Inc. | Protease-cleavable substrates and methods of use thereof |
WO2024030845A1 (en) | 2022-08-01 | 2024-02-08 | Cytomx Therapeutics, Inc. | Protease-cleavable moieties and methods of use thereof |
WO2024030843A1 (en) | 2022-08-01 | 2024-02-08 | Cytomx Therapeutics, Inc. | Protease-cleavable moieties and methods of use thereof |
WO2024030847A1 (en) | 2022-08-01 | 2024-02-08 | Cytomx Therapeutics, Inc. | Protease-cleavable moieties and methods of use thereof |
WO2024030829A1 (en) | 2022-08-01 | 2024-02-08 | The United States Of America, As Represented By The Secretary, Department Of Health And Human Services | Monoclonal antibodies that bind to the underside of influenza viral neuraminidase |
WO2024039672A2 (en) | 2022-08-15 | 2024-02-22 | Dana-Farber Cancer Institute, Inc. | Antibodies against msln and methods of use thereof |
WO2024039670A1 (en) | 2022-08-15 | 2024-02-22 | Dana-Farber Cancer Institute, Inc. | Antibodies against cldn4 and methods of use thereof |
WO2024040114A2 (en) | 2022-08-18 | 2024-02-22 | BioLegend, Inc. | Anti-axl antibodies, antigen-binding fragments thereof and methods for making and using the same |
WO2024059165A1 (en) | 2022-09-15 | 2024-03-21 | Alnylam Pharmaceuticals, Inc. | 17b-hydroxysteroid dehydrogenase type 13 (hsd17b13) irna compositions and methods of use thereof |
Family Cites Families (17)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
US3993754A (en) * | 1974-10-09 | 1976-11-23 | The United States Of America As Represented By The United States Energy Research And Development Administration | Liposome-encapsulated actinomycin for cancer chemotherapy |
US4426330A (en) * | 1981-07-20 | 1984-01-17 | Lipid Specialties, Inc. | Synthetic phospholipid compounds |
US4534899A (en) * | 1981-07-20 | 1985-08-13 | Lipid Specialties, Inc. | Synthetic phospholipid compounds |
US4501728A (en) * | 1983-01-06 | 1985-02-26 | Technology Unlimited, Inc. | Masking of liposomes from RES recognition |
JPS60100516A (en) * | 1983-11-04 | 1985-06-04 | Takeda Chem Ind Ltd | Preparation of sustained release microcapsule |
US4885172A (en) * | 1985-06-26 | 1989-12-05 | The Liposome Company, Inc. | Composition for targeting, storing and loading of liposomes |
IE58981B1 (en) * | 1985-10-15 | 1993-12-15 | Vestar Inc | Anthracycline antineoplastic agents encapsulated in phospholipid micellular particles |
US4797285A (en) * | 1985-12-06 | 1989-01-10 | Yissum Research And Development Company Of The Hebrew University Of Jerusalem | Lipsome/anthraquinone drug composition and method |
GB8601100D0 (en) * | 1986-01-17 | 1986-02-19 | Cosmas Damian Ltd | Drug delivery system |
US4920016A (en) * | 1986-12-24 | 1990-04-24 | Linear Technology, Inc. | Liposomes with enhanced circulation time |
US4837028A (en) * | 1986-12-24 | 1989-06-06 | Liposome Technology, Inc. | Liposomes with enhanced circulation time |
US4863739A (en) * | 1987-05-19 | 1989-09-05 | Board Of Regents, The University Of Texas System | Liposome compositions of anthracycline derivatives |
JPH0696636B2 (en) * | 1988-03-30 | 1994-11-30 | 富士写真フイルム株式会社 | 2,4-bis (o-methoxypolyethylene glycol) -6-cholesteryl-S-triazine compound |
JPH0720857B2 (en) * | 1988-08-11 | 1995-03-08 | テルモ株式会社 | Liposome and its manufacturing method |
AU616040B2 (en) * | 1988-08-11 | 1991-10-17 | Terumo Kabushiki Kaisha | Agents for inhibiting adsorption of proteins on the liposome surface |
GB8824593D0 (en) * | 1988-10-20 | 1988-11-23 | Royal Free Hosp School Med | Liposomes |
US5013556A (en) * | 1989-10-20 | 1991-05-07 | Liposome Technology, Inc. | Liposomes with enhanced circulation time |
-
1989
- 1989-10-20 US US07/425,224 patent/US5013556A/en not_active Expired - Lifetime
-
1990
- 1990-10-19 AU AU66374/90A patent/AU642679B2/en not_active Expired
- 1990-10-19 JP JP3501034A patent/JP2667051B2/en not_active Expired - Lifetime
- 1990-10-19 CA CA002067178A patent/CA2067178C/en not_active Expired - Lifetime
- 1990-10-19 DE DE19675048C patent/DE19675048I2/en active Active
- 1990-10-19 KR KR1019920700919A patent/KR920703013A/en not_active Application Discontinuation
- 1990-10-19 DK DK91900513.2T patent/DK0496835T3/en active
- 1990-10-19 ES ES91900513T patent/ES2071976T3/en not_active Expired - Lifetime
- 1990-10-19 CA CA002067133A patent/CA2067133C/en not_active Expired - Lifetime
- 1990-10-19 DE DE69015207T patent/DE69015207T2/en not_active Revoked
- 1990-10-19 KR KR1019920700918A patent/KR0134982B1/en not_active IP Right Cessation
- 1990-10-19 EP EP90916409A patent/EP0496813B1/en not_active Revoked
- 1990-10-19 JP JP51523890A patent/JP3571335B2/en not_active Expired - Fee Related
- 1990-10-19 AT AT90916409T patent/ATE115401T1/en not_active IP Right Cessation
- 1990-10-19 AT AT91900513T patent/ATE122229T1/en active
- 1990-10-19 AU AU68982/91A patent/AU654120B2/en not_active Expired
- 1990-10-19 WO PCT/US1990/006211 patent/WO1991005546A1/en active IP Right Grant
- 1990-10-19 DE DE69019366T patent/DE69019366T2/en not_active Expired - Lifetime
- 1990-10-19 EP EP91900513A patent/EP0496835B1/en not_active Expired - Lifetime
- 1990-10-19 WO PCT/US1990/006034 patent/WO1991005545A1/en not_active Application Discontinuation
- 1990-10-21 IL IL9606990A patent/IL96069A/en not_active IP Right Cessation
- 1990-10-21 IL IL9607090A patent/IL96070A/en not_active IP Right Cessation
-
1991
- 1991-01-15 US US07/642,321 patent/US5213804A/en not_active Expired - Lifetime
-
1992
- 1992-03-27 NO NO920996A patent/NO304637B1/en not_active IP Right Cessation
- 1992-03-27 NO NO92921213A patent/NO921213L/en unknown
- 1992-04-21 FI FI921764A patent/FI105151B/en active Protection Beyond IP Right Term
- 1992-04-21 FI FI921763A patent/FI921763A0/en not_active Application Discontinuation
-
1995
- 1995-08-09 GR GR950402186T patent/GR3017060T3/en unknown
-
1996
- 1996-12-13 LU LU88854C patent/LU88854I2/en unknown
- 1996-12-18 NL NL960031C patent/NL960031I2/en unknown
-
1997
- 1997-02-05 HK HK14097A patent/HK14097A/en not_active IP Right Cessation
- 1997-03-17 JP JP9063661A patent/JP2889549B2/en not_active Expired - Lifetime
-
1999
- 1999-02-23 NO NO1999003C patent/NO1999003I1/en unknown
-
2001
- 2001-01-11 JP JP2001004291A patent/JP3921050B2/en not_active Expired - Fee Related
Also Published As
Similar Documents
Publication | Publication Date | Title |
---|---|---|
CA2067178C (en) | Solid tumor treatment method and composition | |
US6027726A (en) | Glycosylated protein-liposome conjugates and methods for their preparation | |
JP4555569B2 (en) | Lipid carrier composition having enhanced blood stability | |
JP5117648B2 (en) | Cationic PEG lipids and methods of use. | |
US7108863B2 (en) | Liposome composition for improved intracellular delivery of a therapeutic agent | |
DE60122304T2 (en) | LIPIDEN BASED SYSTEM FOR TARGETED ADMINISTRATION OF DIAGNOSTIC ACTIVE SUBSTANCES | |
AU2002305094A1 (en) | Liposome composition for improved intracellular delivery of a therapeutic agent | |
JP2001510451A (en) | Ion carrier carrying weakly basic drug-liposome in the middle | |
CA2540621A1 (en) | Enhanced drug delivery | |
US5780054A (en) | Methods for increasing the circulation half-life of protein-based therapeutics | |
AU780194B2 (en) | Method of regulating leakage of drug encapsulated in liposomes | |
EP1731172A1 (en) | Liposome preparation | |
US20030113369A1 (en) | Liposomes with enhanced circulation time and method of treatment | |
US20010051183A1 (en) | Liposomes with enhanced circulation time and method of treatment | |
WO2003022250A2 (en) | Unilamellar vesicles stabilized with short chain hydrophilic polymers | |
KR100768265B1 (en) | Heparin coated liposomes to prolong circulation time in bloodstream and preparation method thereof | |
Nallamothu | Development and evaluation of a tumor vasculature targeted liposome delivery system for a novel anti-vascular agent, combretastatin A4 |
Legal Events
Date | Code | Title | Description |
---|---|---|---|
EEER | Examination request | ||
MKEX | Expiry |