CA2597596A1 - Liquid formulations of rapamycin for intraocular delivery - Google Patents

Abstract

Diseases and conditions associated with tissues of the body, including but not limited to tissues in the eye, can be effectively treated, prevented, inhibited, onset delayed, or regression caused by administering therapeutic agents to those tissues. Described herein are liquid formulations which deliver a variety of therapeutic agents, including but not limited to rapamycin, to a subject for an extended period of time; liquid formulations which form a non-dispersed mass when placed in an aqueous medium of a subject;
non- dispersed mass-forming liquid formulations which form a gel or gel-like substance in an aqueous medium; liquid formulations, comprising a therapeutic agent and a plurality of polymers; and methods for delivering therapeutic agents to a subject for an extended period of time using the liquid formulations. The liquid formulation may be placed in an aqueous medium of a subject, including but not limited to via intraocular or periocular administration, or placement proximate to a site of a disease or condition to be treated in a subject. A method may be used to administer rapamycin to treat or prevent angiogenesis, choroidal neovascularization, or age-related macular degeneration, or wet age-related macular degeneration in a subject. The liquid formulations may comprise rapamycin or other therapeutic agents.

Description

LIQUID FORMULATIONS FOR TREATMENT OF DISEASES OR
CONDITIONS

Inventors: Sreenivasu MUDUMBA, Philippe JM DOR, Thierry NIVAGGIOLI, David A. WEBER, Sidiq FAROOQ
FIELD
[0001] Described herein are liquid formulations for treatment, prevention, inhibition, delaying onset of, or causing regression of a disease or condition by delivery of therapeutic agents to a subject, including but not limited to a human subject, including but not limited to the treatment of age-related macular degeneration ("AMD") by delivery of a liquid formulation comprising a therapeutic agent, including but not limited to rapamycin (sirolimus), to the eye of a subject, including but not limited to a human subject.
Nonlimiting exainples of liquid formulations include solutions, suspensions, and in situ gelling formulations.

CROSS-REFERENCES TO RELATED APPLICATIONS

[0002] The present application is related to and claims priority from U.S.
Provisional Patent Application Serial No. 60/664,040 titled "Liquid Formulations For Treatment Of Diseases Or Conditions," filed Mar. 21, 2005, U.S. Provisional Patent Application Serial No. 60/664,306 titled "In Situ Gelling Formulations And Liquid Formulations For Treatment of Diseases Or Conditions," filed Mar. 21, 2005, U.S. Provisional Patent Application Serial No. 60/651,790, titled "Formulations For Ocular Treatment,"
filed Feb.
9, 2005, each of which is incorporated herein by reference in its entirety for all purposes.

BACKGROUND

[0003] The retina of the eye contains the cones and rods that detect light. In the center of the retina is the macula lutea, which is about 1/3 to 1/2 cm in diameter.
The macula provides detailed vision, particularly in the center (the fovea), because the cones are higher in density. Blood vessels, ganglion cells, inner nuclear layer and cells, and the ---- --- -plexiform layers are all displaced to one side (rather than resting above the cones), thereby allowing light a more direct path to the cones.

100041 Under the retina are the choroid, comprising a collection of blood vessels embedded within a fibrous tissue, and the deeply pigmented epithelium, which overlays the choroid layer. The choroidal blood vessels provide nutrition to the retina (particularly its visual cells).
[0005] There are a variety of retinal disorders for which there is currently no treatment or for which the current treatment is not optimal. Retinal disorders such as uveitis (an inflammation of the uveal tract: iris, ciliary body, and choroid), central retinal vein occlusive diseases (CRVO), branch retinal venous occlusion (BRVO), macular degeneration, macular edema, proliferative diabetic retinopathy, and retinal detachment generally are all retinal disorders that are difficult to treat with conventional therapies.
[0006] Age-related macular degeneration (AMD) is the major cause of severe visual loss in the United States for individuals over the age of 60. AMD occurs in either an atrophic or less commonly an exudative form. The atrophic form of AMD is also called "dry AMD," and the exudative form of AMD is also called "wet AMD."

[0007] In exudative AMD, blood vessels grow from the choriocapillaris through defects in Bruch's membrane, and in some cases the underlying retinal pigment epithelium.
Organization of serous or hemorrhagic exudates escaping from these vessels results in fibrovascular scarring of the macular region with attendant degeneration of the neuroretina, detachment and tears of the retinal pigment epithelium, vitreous hemorrhage and permanent loss of central vision. This process is responsible for more than 80% of cases of significant visual loss in subjects with AMD. Current or forthcoming treatments include laser photocoagulation, photodynamic therapy, treatment with VEGF
antibody fragments, treatment with pegylated aptamers, and treatment with certain small molecule agents.
[00081 Several studies have recently described the use of laser photocoagulation in the treatment of initial or recurrent neovascular lesions associated with AMD
(Macular Photocoagulation Study Groups (1991) in Arch. Ophtlaal. 109:1220; AYCIa.
Ophthal.
109:1232; Arch. Ophtlzal. 109:1242). Unfortunately, AMD subjects with subfoveal lesions subjected to laser treatment experienced a rather precipitous reduction in visual acuity (mean 3 lines) at 3 months follow-up. Moreover, at two years post-treatment treated eyes had only marginally better visual acuity than their untreated counterparts (means of 20/320 and 20/400, respectively). Another drawback of the procedure is that vision after surgery is imtnediately worse.

[0009] Photodynamic therapy (PDT) is a fonn of phototherapy, a term encompassing all treatments that use light to produce a beneficial reaction in a subject.
Optimally, PDT
destroys unwanted tissue wllile sparing normal tissue. Typically, a compound called a photosensitizer is administered to the subject. Usually, the photosensitizer alone has little or no effect on the stibject. When light, often from a laser, is directed onto a tissue containing the photosensitizer, the photosensitizer is activated and begins destroying targeted tissue. Because the light provided to the stibject is confined to a particularly targeted area, PDT can be used to selectively target abnormal tissue, thus sparing surrounding healthy tissue. PDT is currently used to treat retinal diseases such as AMD.
PDT is ctirrently the mainstay of treatment for subfoveal choroidal neovascularization in subjects with AMD (Photodynamic Therapy for Subfoveal Choroidal Neovascularization in Age Related Macular Degeneration with Verteporfin (TAP Study Group) Arch Ophtlaalnaol. 1999 117:1329-1345.
[0010] Choroidal neovascularization (CNV) has proven to be recalcitrant to treatnlent in most cases. Conventional laser treatment can ablate CNV and help to preserve vision in selected cases not involving the center of the retina, but this is limited to only about 10% of the cases. Unfortunately, even with successful conventional laser photocoagulation, the neovascularization recurs in about 50-70% of eyes (50%
over 3 years and >60% at 5 years). (Macular Photocoagulation Study Group, Arch.
Ophthalinol.
204:694-701 (1986)). In addition, many subjects who develop CNV are not good candidates for laser therapy because the CNV is too large for laser treatment, or the location cannot be determined so that the physician cannot accurately aim the laser.
Photodynamic therapy, although utilized in up to 50% of new cases of subfoveal CNV has only marginal benefits over natural history, and generally delays progression of visual loss rather than improving vision which is already decreased secondary to the subfoveal lesion.
PDT is neither preventive or definitive. Several PDT treatments are usually required per subject and additionally, certain subtypes of CNV fare less well than others.

[0011] Thus, there remains a long-felt need for methods, compositions, and formulations that may be tised to optimally prevent or significantly inhibit choroidal neovascularization and to prevent and treat wet AMD.
[0012] hi addition to AMD, choroidal neovascularization is associated with such retinal disorders as presumed ocular histoplasmosis syndrome, myopic degeneration, angioid streaks, idiopathic central serous chorioretinopathy, inflammatory conditions of the retina and or choroid, and ocular trauma. Angiogenic damage associated with neovascularization occurs in a wide range of disorders including diabetic retinopathy, venous occlusions, siclcle cell retinopathy, retinopathy of prematurity, retinal detachment, ocular ischemia and trauma.
[0013] Uveitis is another retiiial disorder that has proven difficult to treat using existing therapies. Uveitis is a geiieral tenn that indicates an inflammation of any component of the uveal tract. The uveal tract of the eye consists of the iris, ciliary body, and choroid.
Inflainmation of the overlying retina, called retinitis, or of the optic nerve, called optic neuritis, may occur with or without accompanying uveitis.
[0014] Uveitis is most commonly classified anatomically as anterior, intermediate, posterior, or diffuse. Posterior uveitis signifies any of a number of forms of retinitis, choroiditis, or optic neuritis. Diffuse uveitis implies inflammation involving all parts of the eye, including anterior, intermediate, and posterior structures.
[0015] The symptoms and signs of uveitis may be subtle, and vary considerably depending on the site and severity of the inflammation. Regarding posterior uveitis, the most common symptoms include the presence of floaters and decreased vision.
Cells in the vitreous humor, white or yellow-white lesions in the retina and/or underlying choroid, exudative retinal detachments, retinal vasculitis, and optic nerve edema may also be present in a subject suffering from posterior uveitis.
[0016] Ocular complications of uveitis may produce profound and irreversible loss of vision, especially when unrecognized or treated improperly. The most frequent complications of posterior uveitis include retinal detachment;
neovascularization of the retina, optic nerve, or iris; and cystoid macular edema.
[0017] Macular edema (ME) can occur if the swelling, leaking, and hard exudates noted in background diabetic retinopathy (BDR) occur within the macula, the central 5% of the retina most critical to vision. Background diabetic retinopathy (BDR) typically consists of retinal microaneurisms that result from changes in the retinal microcirculation. These microaneurisms are usually the earliest visible change in retinopathy seen on exam with an ophthalmoscope as scattered red spots in the retina where tiny, weakened blood vessels have ballooned out. The ocular findings in background diabetic retinopathy progress to cotton wool spots, intraretinal hemorrhages, leakage of fluid from the retinal capillaries, and retinal exudates. The increased vascular permeability is also related to elevated levels of local growth factors such as vascular endothelial growth factor. The macula is rich in cones, the nerve endings that detect color and upon which daytime vision depends. When increased retinal capillary permeability effects the macula, blurring occurs in the middle or just to the side of the central visual field, rather like looking through cellophane.

Visual loss may progress over a period of months, and can be very annoying because of the inability to focus clearly. ME is a commoi7 cause of severe visual impairment.
[0018] There have been many attempts to treat CNV and its related diseases and conditions, as well as other conditions such as macular edema and chronic inflammation, with pharmaceuticals. For example, use of rapamycin to inhibit CNV and wet AMD
has been described in U.S. Application No. 10/665,203, which is incorporated herein by reference in its entirety. The use of rapamycin to treat inflammatory diseases of the eye has been described in US patent number 5,387,589, titled Method of Treating Ocular Inflammation, with inventor Prassad Kulkarni, assigned to University of Louisville Research Foundation, the contents of which is incorporated herein in its entirety.

[0019] Particularly for chronic diseases, including those described herein, there is a great need for long acting methods for delivering therapeutic agents to the eye, such as to the posterior segment to treat CNV in such diseases as AMD, macular edema, proliferative retinopathies, and chronic inflammation. Formulations with extended delivery of therapeutic agent are more comfortable and convenient for a subject, due to a diminished frequency of ocular injections of the therapeutic agent.
[0020] Direct delivery of therapeutic agents to the eye rather than systemic administration may be advantageous because the therapeutic agent concentration at the site of action is increased relative to the therapeutic agent concentration in a subject's circulatory system. Additionally, therapeutic agents may have undesirable side effects when delivered systemically to treat posterior segment disease. Thus, localized drug delivery may promote efficacy while decreasing side effects and systemic toxicity.

SUMMARY
[0021] The methods, coinpositions, and liquid formulations described herein allow delivery of a therapeutic agent to a subject, including but not limited to a human subject or to the eye of a subject. Described herein are methods, compositions, and liquid formulations for delivering a variety of therapeutic agents for extended periods of time - - --- - - -- - -----which can be used for the treatment, prevention, inhibition, delaying onset of, or causing regression of a number of conditions or diseases, including but not limited to diseases or conditions of the eye. The liquid formulations include, without limitation, solutions, suspensions, and in situ gelling formulations.
[0022] Described herein are methods, compositions and liquid formulations for administering to a htiman subject an amount of rapainycin effective to treat, prevent, inhibit, delay onset of, or cause regression of wet AMD.
[0023] As described in further detail in the Detailed Description section, the methods, compositions and liquid formulations may also be used for delivery to a subject, including but not limited to a human subject or to the eye of a human subject of therapeutically effective amounts of rapamycin for the treatment, prevention, inhibition, delaying of the onset of, or causing the regression of wet AMD. In some variations, the methods, compositions, and liquid formulations are used to treat wet AMD. In some variations, the metllods, compositions, and liquid formulations are used to prevent wet AMD.
In some variations, the methods and formulations described herein are used to prevent the transition from dry AMD to wet AMD. The methods, compositions and liquid forinulations may also be used for delivery to a subject, including but not limited to a human subject or to the eye of a subject of therapeutically effective amounts of rapamycin for the treatment, prevention, inhibition, delaying of the onset of, or causing the regression of CNV. In some variations, the methods, compositions and liquid formulations are used to treat CNV. The metlzods, compositions and liquid formulations may also be used for delivery to a subject, including but not limited to a human subject or to the eye of a subject. of therapeutically effective amounts of rapamycin for the treatment, prevention, inhibition, delaying of the onset of, or causing the regression of angiogenesis in the eye. In some variations, the methods, compositions and liquid formulations are used to treat angiogenesis. Other diseases and conditions that may be treated, prevented, inhibited, have onset delayed, or caused to regress using rapamycin are described in the Diseases and Conditions section of the Detailed DescYiption.

[00241 As described in further detail in the Detailed Descf iption, the methods, compositions and liquid formulations may also be used for delivery to a subject, including but not limited to a human subject or to the eye of a subject of therapeutically effective amounts of therapeutic agents other than rapamycin for the treatment, prevention, inhibition, delaying of the onset of, or causing the regression of wet AMD. In some variations, the methods, compositions and liquid formulations are used to treat wet AMD.
Therapeutic agents that may be used are described in detail in the TlaeYapeutic Agents section. Such therapeutic agents include but are not limited to immunophilin binding compounds. Immunophilin binding compounds that may be used include but are not limited to the limus family of compounds described further in the Therapeutic Agents section herein, including rapainycin, SDZ-RAD, tacrolimus, everolimus, pimecrolimus, CCI-779, AP23841, ABT-578, derivatives, analogs, prodrugs, salts and esters thereof.
The methods, compositions and liquid formulations may also be used for delivery to a subject, including but not limited to a human subject or to the eye of a subject of therapeutically effective amounts of therapeutic agents for the treatment, prevention, inhibition, delaying of the onset of, or causing the regression of CNV. In some variations, the methods, compositions and liquid formulations are used to treat CNV. The methods, compositions and liquid formulations may also be used for deliveiy to a subject, including but not limited to a human subject or to the eye of a subject of therapeutically effective amounts of therapeutic agents for the treatment, prevention, inhibition, delaying of the onset of, or causing the regression of angiogenesis in the eye. In some variations, the methods, compositions and liquid formulations are used to treat angiogenesis.
Otlier diseases and conditions that may be treated, prevented, inhibited, have onset delayed, or caused to regress using therapeutic agents other than rapamycin are described in the Diseases and Conditions section of the Detailed Description.

[0025] One liquid formulation described herein comprises a solution that includes a tlierapeutic agent dissolved in a solvent. Generally, any solvent that has the desired effect may be used in which the therapeutic agent dissolves and which can be administered to a subject, including but not limited to a human subject or an eye of a subject.
Generally, any concentration of therapeutic agent that has the desired effect can be used. The formulation in some variations is a solution which is unsaturated, a saturated or a supersaturated solution. The solvent may be a pure solvent or may be a mixture of liquid solvent coinponents. In some variations the solution formed is an in situ gelling formulation. Solvents and types of solutions that may be used are well known to those versed in such drug delivery technologies.
[0026] The liquid formulations described herein may form a non-dispersed mass when placed into a rabbit eye, including but not limited to the vitreous of a rabbit eye. In some variations the non-dispersed mass comprises a gel. In some variations, the liquid formulation comprises a therapeutic agent and a plurality of polymers. In some variations one of the polyiners is polyacrylate or polymethacrylate. In some variations one of the polymers is polyvinylpyrrolidone.

[0027] In some variations, the non-dispersed mass comprises a depot. In some variations, the non-dispersed mass consists of a depot.

[0028] For liquid formulations which form a non-dispersed mass, the non-dispersed mass may generally be any geometry or shape. The non-dispersed mass-forming liquid formulations may, for instance, appear as a compact spherical mass when placed in the vitreous. In some variations the liquid fonnulations described herein form a milky or whitish colored semi-contiguous or semi-solid non-dispersed mass relative to the medium in which it is placed, when placed in the vitreous.
[0029] The liquid forinulations may generally be administered in any volume that has the desired effect. In one method a volume of a liquid formulation is administered to the vitreous and the liquid formulation is less than one half the voluine of the vitreous.
[0030] Routes of administration that may be used to administer a liquid formulation include but are not limited to (1) placement of the liquid formulation by placeinent, including by injection, into a mediuin, including but not limited to an aqueous medium in the body, including but not limited to intraocular or periocular injection; or (2) oral administration of the liquid formulation. The liquid formulation may be administered systemically, including but not limited to the following delivery routes:
rectal, vaginal, infusion, intramuscular, intraperitoneal, intraarterial, intrathecal, intrabronchial, intracisternal, cutaneous, subcutaneous, intradermal, transdermal, intravenous, intracervical, intraabdominal, intracranial, intrapulmonary, intrathoracic, intratracheal, nasal, buccal, sublingual, oral, parenteral, or nebulised or aerosolized using aerosol propellants. In some variations, the liquid formulation is administered subconjunctivally.
In some variations, the liquid formulation is administered intravitreally.

[0031] The liquid formulations described herein may be delivered to any medium of a subject, including but not limited to a human subject, including but not limited to an aqueous medium of a subject.

[0032] One liquid formulation described herein comprises a liquid formulation of rapamycin or other therapeutic agent. The liquid formulations may comprise a solution, suspension, an in situ gelling formulation, or an emulsion. The droplets in the emulsion may generally be of any size, including but not limited to up to about 5,000 nm.

[0033] In some formulations described herein, the liquid formulations may comprise a therapeutic agent including but not limited to rapamycin, and one or more solubilizing agents or solvents. In some variations, the soh.ibilizing agent or solvent is glycerin, DMSO, DMA, N-methylp}nTolidone, ethanol, benzyl alcohol, isopropyl alcohol, polyethylene glycol of various molecular weights, including but not limited to and PEG 400, or propylene glycol or a mixture of one or more thereof.

[0034] In some formulations described herein, the liquid formulation includes hyah.ironic acid.

[0035] The liquid formulations described herein may deliver a therapeutic agent or agents for an extended period of time. One nonlimiting example of such an extended release delivery system is a liquid formulation that delivers a therapeutic agent or agents to a subject, including but not limited to a human subject or to the eye of a subject in an amount sufficient to maintain an amount effective to treat, prevent, inhibit, delay onset of, or cause regression of a disease or condition in a subject for an extended period of time.
In some variations, the liquid formulation is used to treat a disease or condition in a subject, including but not limited to a human subject. In some variations, the liquid formulation delivers the therapeutic agent for at least about one, about two, about three, about six, about nine, or about twelve months.

[0036] The liquid formulations described herein may deliver rapamycin or other therapeutic agents for an extended period of time. One nonlimiting example of such an extended release delivery system is a liquid formulation that delivers rapamycin to a subject, including but not limited to a human subject or to the eye of a subject in an amount sufficient to maintain an amount effective to treat, prevent, inhibit, delay onset of, or cause regression of wet age-related macular degeneration for an extended period of time. In some variations, the liquid formulation is used to treat wet age-related macular degeneration for an extended period of time. In some variations, the liquid formulation is used to prevent wet age-related macular degeneration for an extended period of time. In some variations, the liquid formulation is used to prevent transition of dry AMD to wet AMD for an extended period of time. In one nonlimiting example, the liquid formulation delivers the rapamycin to the vitreous, sclera, retina, choroid, macula, or other tissues of a subject, including but not limited to a human subject in an amount sufficient to treat, prevent, inhibit, delay onset of, or cause regression of wet age-related macular degeneration for at least about three, about six, about nine, or about twelve months. In some variations, the level of rapainycin is sufficient to treat AMD. In some variations, the level of rapamycin is sufficient to prevent onset of wet AMD.

[0037] Other extended periods of release are described in the Detailed Description.
BRIEF DESCRIPTION OF THE DRAWINGS
[0038] FIGURES lA-1C schematically depicts formation of a non-dispersed mass, after injection of a liquid formulation into the vitreous of an eye, as it is believed to occur in some variations.

[0039] FIGURE 2 depicts the level of rapamycin in the vitreous (ng/ml), retina choroid (ng/mg), and sclera (ng/mg) of rabbit eyes at 20, 40, 67, and 90 days after subconjunctival injection of a 1.256% solution of rapamycin in water, ethanol, and F127 (Lutrol).

[0040] FIGURE 3 depicts the level of rapamyciil in the vitreous (ng/ml), retina choroid (ng/mg), and sclera (ng/ing) of rabbit eyes at 14, 35, 62, and 85 days after subconjunctival injection of a 5% solution of rapamycin in PEG 400 and ethanol. The level of rapamycin present in the vitreous (ng/ml) is also shown at 2 days after injection.

[0041] FIGURE 4 depicts the level of rapamycin in the vitreous (ng/ml), retina choroid (ng/mg), and sclera (ng/mg) of rabbit eyes at 14, 35, 62, and 90 days after intravitreal injection of a 5% solution of rapamycin in PEG 400 and ethanol. The level of rapamycin present in the vitreous (nglml) is also shown at 2 days after injection.

[0042] FIGURE 5 depicts images of rabbit eyes 8 days after intravitreal injection of 10 gl (Fig. 4A), 20 gl (Fig. 4B), and 40 l (Fig. 4C) of a 6% rapamycin suspension in PEG400.

[0043] FIGURE 6 depicts the level of rapamycin in the vitreous (ng/ml), retina choroid tissues (ng/mg), and sclera (ng/mg) of rabbit eyes at 7, 32, 45, and 90 days after subconjunctival injection of a 4.2% solution of rapamycin in ethanol, PVP K90, PEG 400, and Eudragit RL 100.

[0044] FIGURE 7 depicts the level of rapamycin in the vitreous (ng/ml), retina choroid tissues (ng/mg), and sclera (ng/mg) of rabbit eyes at 14, 42, 63, and 91 days after subconjunctival injection of a 3% suspension of rapamycin in PEG 400.

[0045] FIGURE 8 depicts the level of rapamycin in the vitreous (ng/ml), retina choroid tissues (ng/mg) and sclera (ng/mg) of rabbit eyes_at_ 14, 42,63,_ and 91 days after ,.__ intravitreal injection of a 3% suspension of rapamycin in PEG 400, and in the vitreous at 63 and 91 days after injection.

[0046] FIGURE 9 depicts the level of rapamycin in the vitreous (ng/ml), retina choroid tissues (ng/mg), and sclera (ng/mg) of rabbit eyes at 14, 42, 63, and 91 days after stibconjunctival injection of a 2% soltition of rapainycin in ethanol and PEG
400.
[0047] FIGURE 10 depicts the level of rapamycin in the retina choroid tissues (ng/mg) and sclera (ng/mg) of rabbit eyes at 14, 42, 63, and 91 days after intravitreal injection of a 2% solution of rapamycin in ethanol and PEG 400.
[0048] FIGURE 11 depicts the level of rapamycin in the vitreous (ng/ml) of rabbit eyes at 63 and 91 days after intravitreal injection of a 2% solution of rapamycin in ethanol and PEG 400.
[0049] FIGURE 12 depicts the level of rapamycin in the vitreous (ng/ml) of rabbit eyes at 5, 30, 60, 90, and 120 days after subconjunctival injection of 20 l, 40 l, and 60 l doses of a 2% solution of rapamycin in ethanol and PEG 400.
[00501 FIGURE 13 depicts the level of rapamycin in the retina choroid tissues (ng/mg) of rabbit eyes at 5, 30, 60, 90, and 120 days after subconjunctival injection of 20 1, 40 l, and 60 l doses of a 2% solution of rapainycin in ethanol and PEG 400.

[0051] FIGURE 14 depicts the level of rapamycin in the vitreous (ng/ml) of rabbit eyes at 5, 30, 60, 90, and 120 days after intravitreal injection of 20 l and 40 .l doses of a 2%
solution of rapamycin in ethanol and PEG 400 and of a 100 l dose of a 0.4 %
rapamycin solution in ethanol and PEG 400.
[0052] FIGURE 15 depicts the level of rapamycin in the retina choroid tissues (ng/mg) of rabbit eyes at 5, 30, 60, 90, and 120 days after intravitreal injection of 20 l and 40 l doses of a 2% solution of rapamycin in ethanol and PEG 400 and of a 100 l dose of a 0.4 % rapamycin solution in ethanol and PEG 400.
[0053] FIGURE 16 depicts the level of rapamycin in the vitreous (ng/ml) of rabbit eyes at 5 and 14 days after subconjunctival injection of a single 10 l dose, a single 60 l dose, two 30 l doses, and three 30 l doses of a 2% solution of rapamycin in ethanol and PEG
400.
[0054] FIGURE 17 depicts the level of rapamycin in the retina choroid tissues (ng/nig) of rabbit eyes at 5 and 14 days after subconjunctival injection of a single 10 l dose, a single 60 l dose, two 30 l doses, and three 30 l doses of a 2% solution of rapamycin in __ ---_-_------ ethanol and PEG 400-.

[0055] FIGURE 18 depicts the level of rapamycin in the vitreous (ng/ml) of rabbit eyes at 5, 14, and 30 days after subconjunctival injection of a single 10 }.il dose, a single 30 Ft1 dose, and three 30 l doses of a 3% suspension of rapamycin in PEG 400.

[0056] FIGURE 19 depicts the level of rapamycin in the retina choroid tissues (ng/mg) of rabbit eyes at 5, 14, and 30 days after subconjunctival injection of a single 101.d dose, a single 30 l dose, and three 30 l doses of a 3% suspension of rapamycin in PEG 400.
[0057] FIGURE 20 depicts the level of rapamycin in the retina choroid tissues (ng/zng) of rabbit eyes at 5, 30, and 90 days after intravitreal injection of 10 l of a 0.2% solcttion of rapamycin in ethanol and PEG 400, of 10 1 of a 0.6% solution of rapamycin in ethanol and PEG 400, and of 10 p.1 of a 2% solution of rapamycin in ethanol and PEG
400.

[0058] FIGURE 21 depicts the level of rapamycin in the vitreous (ng/ml) of rabbit eyes at 5, 30, and 90 days after intravitreal injection of 10 l of a 0.2%
soh.ition of rapamycin in ethanol and PEG 400, of 10 l of a 0.6% solution of rapamycin in etlianol and PEG
400, and of 10 l of a 2% solution of rapamycin in ethanol and PEG 400.

[0059] FIGURE 22 depicts the level of rapamycin in the aqueous humor (ng/ml) of rabbit eyes, the cornea (ng/mg), and the retina choroid tissues (ng/mg) at 1, 4, 7, 11, 14, 21, 28, 35, 54, and 56 days after subconjunctival injection of 40 l of a 2%
solution of rapamycin in ethanol and PEG 400.

DETAILED DESCRIPTION
[0060] Described herein are compositions, liquid fonnulations and methods relating to delivery of therapeutic agents to a subject, including but not limited to a human subject or to the eye of a subject. These compositions, liquid formulations, and methods may be used for the treatment, prevention, inhibition, delaying onset of, or causing regression of diseases and conditions of the eye including but not limited to diseases or conditions of the posterior segn2ent, including but not limited to choroidal neovascularization; macular degeneration; age-related macular degeneration, including wet AMD and dry AMD;
retinal angiogenesis; chronic uveitis; and other retinoproliferative conditions. In some variations, the compositions, liquid formulations, and methods are used for the treatment of the aforementioned diseases or conditions of the eye.

[00611 Herein are described (1) the therapeutic agents that may be delivered to a _subject, includingbut -not--Iimited to a human subject-or an-eye of a subject-using-the compositions,liquid formulations, and methods described herein, (2) the diseases and conditions that may be treated, prevented, inliibited, onset delayed, or regression caused by delivery of the therapeutic agents, (3) liquid formulations that may be used to deliver the therapeutic agents, (4) routes of administration for deliveiy of the liquid formulations, (5) extended delivery of therapeutic agents including but not limited to rapamycin, and (6) description of the treatment of CNV and wet AMD by delivery of rapamycin to a subject, including but not limited to a human subject or to the eye of a subject for an extended period of time using the described compositions and liquid formulations.

[0062] The term "about," as used herein, refers to the level of accuracy that is obtained when the methods described herein, such as the methods in the examples, are used.
However, by "about" a certain amount of a component of a formulation is meant 90-110%
of the amount stated.

Therapeutic Agefats [0063] Most generally, any compounds and coinpositions currently known or yet to be discovered that are useful in treating, preventing, inhibiting, delaying the onset of, or causing the regression of the diseases and conditions described herein may be therapeutic agents for use in the compositions, liquid formulations, and methods described herein.
[0064] Therapeutic agents that may be used include compounds that act by binding members of the iinmunophilin family of cellular proteins. Such compounds are known as "immunophilin binding compounds." hnmunophilin binding compounds include but are not limited to the "limus" family of compounds. Examples of limus compounds that may be used include but are not limited to cyclophilins and FK506-binding proteins (FKBPs), including sirolimus (rapamycin) and its water soluble analog SDZ-RAD
(Novartis), TAFA-93 (Isotechnika), tacrolimus, everolimus, RAD-001 (Novartis), pimecrolimus, temsirolimus, CCI-779 (Wyeth), AP23841 (Ariad), AP23573 (Ariad), and ABT-578 (Abbott Laboratories). Limus compound analogs and derivatives that may be used include but are not limited to the compounds described in U.S. Patents 5,527,907;

6,376,517; and 6,329,386 and U.S. Patent Application No. 09/950,307, each of which is incorporated herein by reference in their entirety. Therapeutic agents also include analogs, prodrugs, salts and esters of limus compounds.
[0065] The terms rapamycin, rapa, and sirolimus are used interchangeably herein.
__ _----[0066] Other rapamycin derivatives that may be used include, without limitation, 7-epi-rapamycin, 7-thiomethyl-rapamycin, 7-epi-trimethoxyphenyl-rapamycin, 7-epi-thiomethyl-rapamycin, 7-deinethoxy-rapamycin, 32-demethoxy-rapamycin, 2-desmethyl-rapaniycin, mono- and di-ester derivatives of rapamycin, 27-oximes of rapamycin; 42-oxo analog of rapamycin; bicyclic rapamycins; rapamycin dimers; silyl ethers of rapamycin;
rapamycin arylsulfonates and sulfamates, mono-esters and di-esters at positions 31 and 42, 30-demethoxy rapamycin, and other derivatives described in Vezina et al., "Rapamycin (AY-22,989), A New Antifiingal Antibiotic. I. Taxonoiny Of The Producing Streptomycete And Isolation Of The Active Principle" J. Antibiot. (Tokyo) 28:721-726 (1975); Sehgal et al., "Rapamycin (AY-22,989), A New Antiftingal Antibiotic.
II.
Ferinentation, Isolation And Characterization" J. Antibiot. (Tokyo) 28:727-732 (1975);
Sehgal et al., "Demethoxyrapamycin (AY-24,668), A New Antifungal Antibiotic"
J.
Antibiot. (Tokyo) 36:351-354 (1983); atid Paiva et al., "Incorporation Of Acetate, Propionate, And Methionine Into Rapamycin By Streptomycetes hygroscopicus" J
Nat Prod 54:167-177 (1991), WO 92/05179, EP 467606, Caufield et al., "Hydrogenated Rapamycin Derivatives" U.S. Pat. No. 5,023,262; Kao et al., "Bicyclic Rapamycins" U.S.
Pat. No. 5,120,725; Kao et al., "Rapamycin Dimers" U.S. Pat. No. 5,120,727;
Failli et al., "Silyl Ethers Of Rapamycin" U.S. Pat. No. 5,120,842; Failli et al., "Rapamycin Sulfonates And 42-(N-carboalkoxy) Sulfalnates Useftil As Immunosuppressive Agents"
U.S. Pat. No. 5,177,203; Nicolaou et al., "Total Synthesis Of Rapamycin" J.
Am. Chem.
Soc. 115: 4419-4420 (1993); Romo et al, "Total Synthesis Of (-) Rapamycin Using An Evans-Tishchenko Fragment Coupling" J. Am. Chem. Soc. 115:7906-7907 (1993);
and Hayward et al, "Total Synthesis Of Rapamycin Via A Novel Titanium-Mediated Aldol Macrocyclization Reaction" J. Am. Chem. Soc., 115:9345-9346 (1993), each of which is incorporated herein by reference in its entirety. , [0067] The limus family of compounds may be used in the compositions, liquid formulations and methods for the treatment, prevention, inhibition, delaying the onset of, or causing the regression of angiogenesis-mediated diseases and conditions of the eye, including choroidal neovascularization. The limus family of compounds may be used to prevent, treat, inhibit, delay the onset of, or cause regression of AMD, including wet AMD. Rapamycin and rapamycin derivatives and analogs may be used to prevent, treat, inhibit, delay the onset of, or cause regression of angiogenesis-mediated diseases and conditions of the eye, including choroidal neovascularization. Rapamycin may be used to prevent, treat, inhibit, delay the onset of, or cause regression of AMD, including wet AMD. In some variations, a member of the limus family of compounds or rapamycin is used to treat wet AMD or angiogenesis-mediated diseases and conditions of the eye including choroidal neovascularization.
[0068] Other therapeutic agents that may be used include those disclosed in the following patents and publications, the contents of each of which is incorporated herein by reference in its eiitirety: PCT publication WO 2004/027027, published April 1, 2004, titled Method of inhibiting choroidal neovascularization, assigned to Tnistees of the University of Pennsylvania; US patent number 5,387,589, issued Febniary 7, 1995, titled Method of Treating Ocular Inflammation, with inventor Prassad Kullcarni, assigned to University of Louisville Research Foundation; US patent number 6,376,517, issued April 23, 2003, titled Pipecolic acid derivatives for vision and memory disorders, assigned to GPI NIL Holdings, Inc; PCT publication WO 2004/028477, published April 8, 2004, titled Method subretinal adininistration of therapeutics including steroids:
method for localizing pharmadynamic action at the choroid and retina; and related methods for treatment and or prevention of retinal diseases, assigned to Innorx, Inc; US
patent number 6,416,777, issued July 9, 2002, titled Ophthalmic drn.ig delivery device, assigned to Alcon Universal Ltd; US patent number 6,713,081, issued March 30, 2004, titled Ocular therapeutic agent delivery device and methods for making and using such devices, assigned to Department of Health and Human Services; US patent number 5,100,899, issued Mar. 31, 1992, titled Methods of inhibiting transplant rejection in mammals using rapamycin and derivatives and prodrugs thereof .
[0069] Other therapeutic agents that may be used include pyrrolidine, dithiocarbamate (NFicB inhibitor); squalamine; TPN 470 analogue and fumagillin; PKC (protein kinase C) inhibitors; Tie-1 and Tie-2 kinase inhibitors; inhibitors of VEGF receptor kinase;
proteosome inhibitors such as VelcadeTM (bortezomib, for injection;
ranibuzumab (LucentisTM) and other antibodies directed to the same target; pegaptanib (MacugenTM);
vitronectin receptor antagonists, such as cyclic peptide antagonists of vitronectin receptor-type integrins; a-v/(3-3 integrin antagonists; a-v/(3-l integrin antagonists;
thiazolidinediones such as rosiglitazone or troglitazone; interferon, including y-interferon or interferon targeted to CNV by use of dextran and metal coordination;
pigment epithelium derived factor (PEDF); endostatin; angiostatin; tumistatin;
canstatin;
anecortave acetate; acetonide; triamcinolone; tetrathiomolybdate; RNA
silencin_or RNA
interference (RNAi) of angiogenic factors, including ribozymes that target VEGF
expression; AccutaneTM (13-cis retinoic acid); ACE inhibitors, including but not limited to quinopril, captopril, and perindozril; inhibitors of mTOR (mammalian target of rapamycin); 3-aminothalidomide; pentoxifylline; 2-methoxyestradiol;
colchicines; AMG-1470; cyclooxygenase inhibitors such as nepafenac, rofecoxib, diclofenac, rofecoxib, NS398, celecoxib, vioxx, and (E)-2-alkyl-2(4-methanesulfonylphenyl)-1-phenylethene; t-RNA synthase modulator; metalloprotease 13 inhibitor; acetylcholinesterase iilllibitor;
potassium channel blockers; endorepellin; purine analog of 6-thioguanine;
cyclic peroxide ANO-2; (recombinant) arginine deiminase; epigallocatechin-3-gallate;
cerivastatin;
analogues of suramin; VEGF trap molecules; apoptosis inhibiting agents;
VisudyneTM, snET2 and other photo sensitizers, which may be used with photodynamic therapy (PDT);
inhibitors of hepatocyte growth factor (antibodies to the growth factor or its receptors, small molecular inhibitors of the c-met tyrosine kinase, tn.incated versions of HGF e.g.
NK4).

[0070) Other therapeutic agents that may be used include anti-inflammatory agents, including, but not limited to nonsteroidal anti-inflammatory agents and steroidal anti-inflammatory agents. In some variations, active agents that may be used in the liquid formulations are ace-inhibitors, endogenous cytokines, agents that influence basement membrane, agents that influence the growth of endothelial cells, adrenergic agonists or blockers, cholinergic agonists or blockers, aldose reductase inhibitors, analgesics, anesthetics, antiallergics, antibacterials, antihypertensives, pressors, antiprotozoal agents, antiviral agents, antifungal agents, anti-infective agents, antitumor agents, antimetabolites, and antiangiogenic agents.

(0071] Steroidal therapeutic agents that may be used include but are not limited to 21-acetoxypregnenolone, alclometasone, algestone, amcinonide, beclomethasone, betamethasone, budesonide, chloroprednisone, clobetasol, clobetasone, clocortolone, cloprednol, corticosterone, cortisone, cortivazol, deflazacort, desonide, desoximetasone, dexamethasone, diflorasone, diflucortolone, difluprednate, enoxolone, fluazacort, flucloronide, flumethasone, flunisolide, fluocinolone acetonide, fluocinonide, fluocortin butyl, fluocortolone, fluorometholone, fluperolone acetate, fluprednidene acetate, fluprednisolone, flurandrenolide, fluticasone propionate, formocortal, halcinonide, halobetasol propionate, halometasone, halopredone acetate, hydrocortamate, hydrocortisone, loteprednol etabonate, mazipredone, medrysone, meprednisone, methylprednisolone, mometasone furoate, paramethasone, prednicarbate, prednisolone, prednisolone 25-diethylamino-acetate, prednisolone sodium phosphate, prednisone, prednival, prednylidene, rimexolone, tixocortol, triamcinolone, triaincinolone acetonide, triamcinolone benetonide, triaincinolone hexacetonide, and any of their derivatives.
[0072] In some variations, cortisone, dexamethasone, fluocinolone, hydrocortisone, methylprednisolone, prednisolone, prednisone, and triamcinolone, or their derivatives, may be used. The liquid fonnulation may include a combination of two or more steroidal therapeutic agents.

[0073] In one nonlimiting example, the steroidal tllerapeutic agents may constitute from about 0.05% to about 50% by weight of the liqtiid formulation. In another nonlimiting example, the steroid constitutes from about 0.05% to about 10%, between about 10% to about 20%; between about 30% to about 40%; or between about 40% to about 50%
by weight of the liquid formulation.

(0074] Other nonlimiting examples of therapeutic agents that may be used include but are not limited to anaesthetics, analgesics, cell transport/mobility impending agents such as colcliicines, vincristine, cytochalasin B and related compounds; carbonic anhydrase iuhibitors such as acetazolamide, methazolamide, dichlorphenamide, diamox and neuroprotectants such as nimodipine and related compounds; antibiotics such as tetracycline, chlortetracycline, bacitracin, neomycin, polymyxin, gramicidin, cephalexin, oxytetracycline, chloramphenicol, rifampicin, ciprofloxacin, aminosides, gentamycin, erythromycin and penicillin, quinolone, ceftazidime, vancomycine iinipeneme;
antifungals such as amphotericin B, fluconazole, ketoconazole and miconazole;
antibacterials such as sulfonamides, sulfadiazine, sulfacetamide, sulfamethizole and sulfisoxazole, nitrofurazone and sodium propionate; antivirals, such as idoxuridine, trifluorothymidine, trifluorouridine, acyclovir, ganciclovir, cidofovir, interferon, DDI, AZT, foscamet, vidarabine, irbavirin, protease inhibitors and anti-cytomegalovirus agents;
antiallergenics such as sodium cromoglycate, antazoline, methapyriline, chlorpheniramine, cetirizine, pyrilamine and prophenpyridamine; synthetic gluocorticoids and mineralocorticoids and more generally hormones forms derivating from the cholesterol metabolism (DHEA, progesterone, estrogens); non-steroidal anti-inflammatories such as salicylate, indomethacin, ibuprofen, diclofenac, flurbiprofen, piroxicam and COX2 inhibitors; antineoplastics such as carmustine, cisplatin, fluorouracil; adriamycin, asparaginase, azacitidine, azathioprine, bleomycin, busulfan, __-.--------carboplatin, carmustine, chlorambucil, cyclophosphamide, cyclosporine, cytarabine, dacarbazine, dactinomycin, daunorubicin, doxorubicin, estramustine, etoposide, etretinate, filgrastin, floxuridine, fludarabine, fluorouracil, florxymesterone, flutamide, goserelin, hydroxyurea, ifosfamide, lettprolide, levamisole, limustine, nitrogen mustard, melphalan, mercaptopurine, methotrexate, mitomycin, mitotane, pentostatin, pipobroznan, plicamycin, procarbazine, sargramostin, streptozocin, tamoxifen, taxol, teniposide, thioguanine, uracil mustard, vinblastine, vincristine and vindesiile;
immunological drugs such as vaccines and immune stiinulants; insulin, calcitonin, parathyroid hormone and peptide and vasopressin hypothalamus releasing factor; beta adrenergic blockers such as timolol, levobunolol and betaxolol; cytokines, interleulcines and growth factors epidermal growth factor, fibroblast growth factor, platelet derived growth factor, transforining growth factor beta, ciliary neurotrophic growth factor, glial derived neurotrophic factor, NGF, EPO, PLGF, brain nerve growth factor (BNGF), vascular endothelial growth factor (VEGF) and monoclonal antibodies or fragments thereof directed against such growth factors; anti-inflammatories such as hydrocortisone, dexamethasone, fluocinolone, prednisone, prednisolone, methylprednisolone, fluorometholone, betamethasone and triamcinolone; decongestants such as phenylephrine, naphazoline and tetrahydrazoline;
miotics asid anti-cholinesterases such as pilocarpine, carbachol, di-isopropyl fluorophosphate, phospholine iodine and demecarium bromide; mydriatics such as atropine sulphate, cyclopentolate, homatropine, scopolamine, tropicamide, eucatropine;
sympatllomimetics such as epinephrine and vasoconstrictors and vasodilators, anticlotting agents such as heparin, antifibrinogen, fibrinolysin, anticlotting activase, antidiabetic agents include acetohexamide, chlorpropamide, glipizide, glyburide, tolazamide, tolbutamide, insulin and aldose reductase inhibitors, hormones, peptides, nucleic acids, saccharides, lipids, glycolipids, glycoproteins and other macromolecules include endocrine horinones such as pituitary, insulin, insulin-related growth factor, thyroid, growth hormones; heat shock proteins; immunological response modifiers such as muramyl dipeptide, cyclosporins, interferons (including alpha-, beta- and gamma-interferons), interleukin-2, cytokines, FK506 (an epoxy-pyrido-oxaazcyclotricosine-tetrone, also known as Tacrolimus), tumor necrosis factor, pentostatin, thymopentin, transforming factor beta2, erythropoetin; antineogenesis proteins (e.g. anti VEGF, interferons), antibodies (monoclonal, polyclonal, humanized, etc.) or antibodies fragments, oligoaptamers, aptamers and gene fragments (oligonucleotides, plasmids, ribozymes, small interference RNA (SiRNA), nucleic acid fragments, peptides), immunomodulators such as endoxan, thalidomide, tamoxifene; antithrombolytic and vasodilator agents such as rtPA, urokinase, plasmin; nitric oxide donors, nucleic acids, dexamethasone, cyclosporin A, azathioprine, brequinar, gusperimus, 6-mercaptoptuine, mizoribine, rapamycin, tacrolimus (FK-506), folic acid analogs (e.g., denopterin, edatrexate, methotrexate, piritrexim, pteropterin, Tomudex@, trimetrexate), purine analogs (e.g., cladribine, fludarabine, 6-mercaptopurine, thiamiprine, thiaguanine), pyrimidine analogs (e.g., ancitabine, azacitidine, 6-azauridine, carmofiir, cytarabine, doxifluridine, ernitefitr, enocitabine, floxuridine, fluorouracil, gemcitabine, tegafur) fluocinolone, triaminolone, anecortave acetate, fluorometholone, medrysone, and prednislone. In some variations the immunosuppressive agent is dexamethasone.
In some variations the iminunosuppressive agent is cyclosporin A.
[0075] In some variations the formulation comprises a combination of one or more therapeutic agents.
[0076] Other nonlimiting examples of therapeutic agents that may be used in the formulations described herein include antibacterial antibiotics, aminoglycosides (e.g., amilcacin, apramycin, arbekacin, bambermycins, butirosin, dibekacin, dihydrostreptomycin, fortimicin(s), gentamicin, isepamicin, kanamycin, micronomicin, neomycin, neomycin undecylenate, netilmicin, paromomycin, ribostamycin, sisomicin, spectinomycin, streptomycin, tobramycin, trospectomycin), amphenicols (e.g., azidamfenicol, chloramphenicol, florfenicol, thiamphenicol), ansamycins (e.g., rifamide, rifampin, rifamycin sv, rifapentine, rifaximin), P-lactams (e.g., carbacephems (e.g., loracarbef), carbapenems (e.g., biapenem, imipenem, meropenem, panipenein), cephalosporins (e.g., cefaclor, cefadroxil, cefamandole, cefatrizine, cefazedone, cefazolin, cefcapene pivoxil, cefclidin, cefdinir, cefditoren, cefepime, cefetamet, cefixime, cefinenoxime, cefodizime, cefonicid, cefoperazone, ceforanide, cefotaxime, cefotiam, cefozopran, cefpimizole, cefpiramide, cefpirome, cefpodoxime proxetil, cefprozil, cefroxadine, cefsulodin, ceftazidime, cefteram, ceftezole, ceftibuten, ceftizoxime, ceftriaxone, cefuroxime, cefuzonam, cephacetrile sodium, cephalexin, cephaloglycin, cephaloridine, cephalosporin, cephalothin, cephapirin sodium, cephradine, pivcefalexin), cephamycins (e.g., cefbuperazone, cefinetazole, cefininox, cefotetan, cefoxitin), monobactams (e.g., aztreonam, carumonam, tigemonam), oxacephems, flomoxef, moxalactam), penicillins (e.g., amdinocillin, amdinocillin pivoxil, amoxicillin, ampicillin, -apalcillin, aspoxicillin, azidocillin, azlocillin, bacainpicillin, benzylpenicillinic acid, benzylpenicillin sodium, carbenicillin, carindacillin, clometocillin, cloxacillin, cyclacillin, dicloxacillin, epicillin, fenbenicillin, floxacillin, hetacillin, lenampicillin, metampicillin, methicillin sodium, mezlocillin, nafcillin sodium, oxacillin, penamecillin, penethamate hydriodide, penicillin g benethamine, penicillin g benzathine, penicillin g benzhydiylamine, penicillin g calcium, penicillin g hydrabamine, penicillin g potassium, penicillin g procaine, penicillin n, penicillin o, penicillin v, penicillin v benzathine, penicillin v liydrabamine, penimepicycline, phenethicillin potassium, piperacillin, pivampicillin, propicillin, quinacillin, sulbenicillin, sultamicillin, talampicillin, temocillin, ticarcillin), ritipenem, lincosamides (e.g., clindamycin, lincomycin), macrolides (e.g., azithromycin, carbomycin, clarithromycin, dirithromycin, erythromycin, erythromycin acistrate, erythromycin estolate, erythromycin glucoheptonate, erythromycin lactobionate, erythromycin propionate, erythromycin stearate, josamycin, leucomycins, midecamycins, miokainycin, oleandomycin, primycin, rok-itamycin, rosaramicin, roxithromycin, spiramycin, troleandomycin), polypeptides (e.g., amphomycin, bacitracin, capreomycin, colistin, enduracidin, enviomycin, fusafungine, gramicidin s, gramicidin(s), mikamycin, polymyxin, pristinamycin, ristocetin, teicoplanin, thiostrepton, tuberactinolnycin, tyrocidine, tyrothricin, vancomycin, viomycin, virginiamycin, zinc bacitracin), tetracyclines (e.g., apicycline, chlortetracycline, clomocycline, demeclocycline, doxycycline, guamecycline, lymecycline, meclocycline, methacycline, minocycline, oxytetracycline, penimepicycline, pipacycline, rolitetracycline, sancycline, tetracycline), and others (e.g., cycloserine, mupirocin, tuberin); synthetic antibacterials, 2.4-Diaminopyrimidines (e.g., brodimoprim, tetroxoprim, trimethoprim), nitrofurans (e.g., furaltadone, furazolium chloride, nifuradene, nifuratel, nifurfoline, nifurpirinol, nifurprazine, nifurtoinol, nitrofurantoin), quinolones and analogs (e.g., cinoxacin, ciprofloxacin, clinafloxacin, difloxacin, enoxacin, fleroxacin, flumequine, grepafloxacin, lomefloxacin, miloxacin, nadifloxacin, nalidixic acid, norfloxacin, ofloxacin, oxolinic acid, pazufloxacin, pefloxacin, pipemidic acid, piromidic acid, rosoxacin, rufloxacin, sparfloxacin, temafloxacin, tosufloxacin, trovafloxacin), sulfonamides (e.g., acetyl sulfamethoxypyrazine, benzylsulfamide, chloramine-b, chloramine-t, dichloramine t, n2-fornnylsulfisomidine, n4-6-d-glucosylsulfanilamide, mafenide, 4'-(methylsulfamoyl)sulfanilanilide, noprylsulfamide, phthalylsulfacetamide, phthalylsulfathiazole, salazosulfadimidine, succinylsulfathiazole, sulfabenzamide, sulfacetamide, sulfachlorpyridazine, sulfachrysoidine, sulfacytine, sulfadiazine, sulfadicramide, sulfadimethoxine, sulfadoxine, sulfaethidole, sulfaguanidine, sulfagua.nol, sulfalene, sulfaloxic acid, sulfameraziiie, sulfaineter, sulfamethazine, sulfamethizole, sulfamethomidine, sulfamethoxazole, sulfamethoxypyridazine, sulfametrole, sulfamidochrysoidine, sulfamoxole, sulfanilamide, 4-sulfanilamidosalicylic acid, n4-sulfanilylsulfanilamide, sulfanilylurea, n-sulfanilyl-3,4-xylamide, sulfanitran, sulfaperine, sulfaphenazole, sulfaproxyline, stilfapyrazine, sulfapyridine, sulfasomizole, sulfasymazine, sulfathiazole, sulfathiourea, sulfatolamide, sulfisomidine, sulfisoxazole) sulfones (e.g., acedapsone, acediasulfone, acetosulfone sodium, dapsone, diathymosulfone, glucosulfone sodium, solasulfone, succisulfone, sulfanilic acid, p-sulfanilylbenzylamine, sulfoxone sodium, thiazolsulfone), and others (e.g., clofoctol, hexedine, methenamine, methenamine anhydromethylene-citrate, methenamine hippurate, methenainine mandelate, methenamine sulfosalicylate, nitroxoline, taurolidine, xibomol), antifungal antibiotics, polyenes (e.g., amphotericin b, candicidin, dennostatin, filipin, fungichromin, hachimycin, hamycin, lucensomycin, mepartricin, natamycin, nystatin, pecilocin, perimycin), azasertne, griseofulvin, oligomycins, neomycin undecylenate, pyrrolnitrin, siccanin, tubercidin, viridin, synthetic antifungals, allylamines (e.g., butenafine, naftifine, terbinafine), imidazoles (e.g., bifonazole, butoconazole, chlordantoin, chlormidazole, cloconazole, clotrimazole, econazole, enilconazole, fenticonazole, flutrimazole, isoconazole, ketoconazole, lanoconazole, miconazole, omoconazole, oxiconazole nitrate, sertaconazole, sulconazole, tioconazole), thiocarbainates (e.g., tolciclate, tolindate, tolnaftate), triazoles (e.g., fluconazole, itraconazole, saperconazole, terconazole), acrisorcin, amorolfine, biphenamine, bromosalicylchloranilide, buclosamide, calcium propionate, chlorphenesin, ciclopirox, cloxyquin, coparaffinate, diamthazole dihydrochloride, exalarnide, flucytosine, halethazole, hexetidine, loflucarban, nifuratel, potassium iodide, propionic acid, pyrithione, salicylanilide, sodiun-i propionate, sulbentine, tenonitrozole, triacetin, ujothion, undecylenic acid, zinc propionate, antineoplastics, antibiotics and analogs (e.g., aclacinomycins, actinomycin fl, anthramycin, azaserine, bleoinycins, cactinomycin, carubicin, carzinophilin, chromomycins, dactinomycin, daunorubicin, 6-diazo-5-oxo-L-norleucine, doxorubicin, epinibicin, idarubicin, menogaril, mitomycins, mycophenolic acid, nogalamycin, olivomycines, peplomycin, pirarubicin, plicamycin, porfiromycin, puromycin, streptonigrin, streptozocin, tubercidin, zinostatin, zorubicin), antimetabolites (e.g. folic acid analogs (e.g., denopterin, edatrexate, methotrexate, piritrexim, pteropterin, Tomudex , trimetrexate), purine analogs (e.g., cladribine, fludarabine, 6-mercaptopurine, thiamiprine, thioguai-une), pyrimidine analogs (e.g., ancitabine, azacitidine, 6-azauridine, carmofur, cytarabine, doxifluridine, emitefur, enocitabine, floxuridine, fluorouracil, gemcitabine, tagafur), antiinflammatory agents, steroidal antiinflaminatory agents, acetoxypregnenolone, alclometasone, algestone, amcinonide, beclomethasone, betamethasone, budesonide, chloroprednisone, clobetasol, clobetasone, clocortolone, cloprednol, corticosterone, cortisone, cortivazol, deflazacort, desonide, desoximetasone, dexamethasone, diflorasone, diflucortolone, difluprednate, enoxolone, fluazacort, flucloronide, flumethasone, flunisolide, fluocinolone acetonide, fluocinonide, fluocortin butyl, fluocortolone, fluorometholone, fluperolone acetate, fluprednidene acetate, fluprednisolone, flurandrenolide, fluticasone propionate, formocortal, halcinonide, halobetasol propionate, halometasone, halopredone acetate, llydrocortamate, hydrocortisone, loteprednol etabonate, mazipredone, medrysone, meprednisone, methylprednisolone, mometasone fiiroate, paramethasone, prednicarbate, prednisolone, prednisolone 25-diethylamino-acetate, prednisolone sodium phosphate, prednisone, prednival, prednylidene, rimexolone, tixocortol, triamcinolone, triamcinolone acetonide, triamcinolone benetonide, and triamcinolone hexacetonide, non-steroidal antiinflammatory agents, aminoarylcarboxylic acid derivatives (e.g., enfenamic acid, etofenamate, flufenamic acid, isonixin, meclofenamic acid, mefenamic acid, niflumic acid, talniflumate, terofenamate, tolfenamic acid), arylacetic acid derivatives (e.g., aceclofenac, acemetacin, alclofenac, amfenac, amtolmetin guacil, bromfenac, bufexamac, cinmetacin, clopirac, diclofenac sodium, etodolac, felbinac, fenclozic acid, fentiazac, glucametacin, ibufenac, indomethacin, isofezolac, isoxepac, lonazolac, metiazinic acid, mofezolac, oxametacine, pirazolac, proglumetacin, sulindac, tiaramide, tolmetin, tropesin, zomepirac), arylbutyric acid derivatives (e.g., bumadizon, butibufen, fenbufen, xenbucin), arylcarboxylic acids (e.g., clidanac, ketorolac, tinoridine), arylpropionic acid derivatives (e.g., alminoprofen, benoxaprofen, bermoprofen, bucloxic acid, carprofen, fenoprofen, flunoxaprofen, flurbiprofen, ibuprofen, ibuproxam, indoprofen, ketoprofen, loxoprofen, naproxen, oxaprozin, piketoprolen, pirprofen, pranoprofen, protizinic acid, suprofen, tiaprofenic acid, ximoprofen, zaltoprofen), pyrazoles (e.g., difenamizole, epirizole), pyrazolones (e.g., apazone, benzpiperylon, feprazone, mofebutazone, morazone, oxyphenbutazone, phenylbutazone, pipebuzone, propyphenazone, ramifenazone, suxibuzone, thiazolinobutazone), salicylic acid derivatives (e.g., acetaminosalol, aspirin, benorylate, bromosaligenin, calcium acetylsalicylate, diflunisal, etersalate, fendosal, gentisic acid, glycol salicylate, imidazole salicylate, lysine acetylsalicylate, mesalamine, morpholine salicylate, 1-naphtllyl salicylate, olsalazine, parsalmide, phenyl acetylsalicylate, phenyl salicylate, salacetamide, salicylamide o-acetic acid, salicylsulfuric acid, salsalate, sulfasalazine), thiazinecarboxamides (e.g., ampiroxicam, droxicam, isoxicam, lomoxicam, piroxicam, teiioxicam), E-acetamidocaproic acid, s-adenosylmethionine, 3-amino-4-hydroxybutyric acid, amixetrine, bendazac, benzydamine, a-bisabolol, bucolome, difenpirainide, ditazol, emorfazone, fepradinol, guaiazulene, nabumetone, nimesulide, oxaceprol, paranyline, perisoxal, proquazone, superoxide dismutase, tenidap, and zileuton.

[0077] The therapeutic agents may also be used in combination with other therapeutic agents and therapies, including but not limited to agents and therapies usefiil for the treatment, prevention, inhibition, delaying onset of, or causing regression of angiogenesis or neovascularization, particularly CNV. In some variations the additional agent or therapy is used to treat regression of angiogenesis or neovascularization, particularly CNV. Non-limiting examples of such additional agents and therapies include pyrrolidine, dithiocarbamate (NFxB inhibitor); squalamine; TPN 470 analogue and fumagillin;
PKC
(protein kinase C) inhibitors; Tie-1 and Tie-2 kinase inhibitors; inhibitors of VEGF
receptor kinase; proteosome inhibitors such as VelcadeTM (bortezomib, for injection;
ranibuzumab (LucentisTM) and other antibodies directed to the same target;
pegaptanib (MacugenTM); vitronectin receptor antagonists, such as cyclic peptide antagonists of vitronectin receptor-type integrins; a-v/(3-3 integrin antagonists; a-v/(3-1 integrin antagonists; thiazolidinediones such as rosiglitazone or troglitazone;
interferon, including y-interferon or interferon targeted to CNV by use of dextran and metal coordination;
pigment epithelium derived factor (PEDF); endostatin; angiostatin; tumistatin;
canstatin;
anecortave acetate; acetonide; triamcinolone; tetrathiomolybdate; RNA
silencing or RNA
interference (RNAi) of angiogenic factors, including ribozymes that target VEGF
expression; AccutaneTM (13-cis retinoic acid); ACE inhibitors, including but not limited to quinopril, captopril, and perindozril; inhibitors of mTOR (mammalian target of rapamycin); 3-aminothalidomide; pentoxifylline; 2-methoxyestradiol;
colchicines; AMG-1470; cyclooxygenase inhibitors such as nepafenac, rofecoxib, diclofenac, rofecoxib, NS398, celecoxib, vioxx, and (E)-2-alkyl-2(4-meth_anesulfony_lphenyl)-1-pheny_lethene;t7 RNA synthase modulator; metalloprotease 13 inhibitor; acetylcholinesterase inhibitor;
potassium channel blockers; endorepellin; purine analog of 6-thioguanine;
cyclic peroxide ANO-2; (recombinant) arginine deiminase; epigallocatechin-3-gallate;
cerivastatin;
analogues of suramin; VEGF trap molecules; inhibitors of hepatocyte growth factor (antibodies to the growth factor or its receptors, small molecular inhibitors of the c-met tyrosine kinase, truncated versions of HGF e.g. NK-4); apoptosis inhibiting agents;
VisudyneTM, snET2 and otlier photo sensitizers with photodynamic therapy (PDT); and laser photocoagulation.

Diseases artd conditiosas that may be treated, pf-everated, iftliibited, ortset delayed, or regressiorz caused [0078] Herein are described diseases and conditions that may be treated, prevented, inhibited, onset delayed, or regression caused using the therapeutic agents and the fonnulations, liquid formutations, and methods described herein. In some variations, the diseases or conditions are treated using the therapeutic agents and the formulations, liquid formulations, and methods described herein. Unless the context indicates otherwise, it is envisioned that the subjects on whom all of the methods of treatment may be performed include, but are not limited to, human subjects.

[0079] Generally, any diseases or condition of the eye susceptible to treatment, prevention, inhibition, delaying the onset of, or causing the regression of using the therapeutic agents and the formulations, liquid formulations and methods described herein may be treated, prevented, inhibited, onset delayed, or regression caused treated or prevented. Examples of diseases or conditions of the eye include, but are not limited to, diseases or conditions associated with neovascularization including retinal and/or choroidal neovascularization.
[0080] Diseases or conditions associated with retinal and/or choroidal neovascularization that can be treated, prevented inhibited, have onset delayed, or be caused to regress using the formulations, liquid formulations, and methods described herein include, but are not limited to, diabetic retinopathy, macular degeneration, wet and dry AMD, retinopathy of prematurity (retrolental fibroplasia), infections causing a retinitis or choroiditis, presumed ocular histoplasmosis, myopic degeneration, angioid streak-s, and ocular trauma. Other non-limiting examples of diseases and conditions of the eye that may be treated, prevented inhibited, have onset delayed, or be caused to regress using the - - - -----_ -------formulations, liquid formulations, and methods described herein include, but are not limited to, pseudoxanthoma elasticum, vein occlusion, artery occlusion, carotid obstructive disease, Sickle Cell anemia, Eales disease, myopia, chronic retinal detachlnent, hyperviscosity syndromes, toxoplasmosis, trauma, polypoidal choroidal vasculopathy, post-laser complications, complications of idiopathic central serous chorioretinopathy, complications of choroidal inflammatory conditions, nibeosis, diseases associated with rubeosis (neovascularization of the angle), neovascular glaucoma, uveitis and chronic uveitis, macular edema, proliferative retinopathies and diseases or conditions caused by the abnormal proliferation of fibrovascular or fibrous tissue, including all forms of proliferative vitreoretinopathy (including post-operative proliferative vitreoretinopathy), whether or not associated with diabetes.

[0081] In some variations, the formulations and phannaceutical formulations described herein are used to prevent or delay onset of a disease or condition of the eye where the subject, including but not limited to a human subject, is at heiglztened risk of developing the disease or condition of the eye. A subject with a heightened risk of developing a disease or condition is a subject with one or more indications that the disease or condition is likely to develop in the particular subject. In some variations the subject with a heightened risk of developing wet AMD is a subject with dry AMD in at least one eye. In some variations the subject with a heightened risk of developing wet AMD in a fellow eye is a subject with wet AMD in the other eye. In some variations, the formulations and pharmaceutical fornnulations described herein are used to prevent or delay onset of CNV
in a subject at heightened risk of developing CNV, including but not limited to prevention or delaying onset of CNV in the fellow eye of a subject, including but not limited to a human subject with AMD in one eye. In some variations, the formulations and pharmaceutical formulations described herein are used to prevent or delay onset of CNV
in the fellow eye of a subject with wet AMD in one eye. In some variations, the formulations and pharmaceutical formulations comprise a limus compound, including but not limited to rapamycin. In some variations the formulations and pharmaceutical fonnulations are adininistered periocularly, including without limitation subconjunctivally, to a human subject with vision of 20/40 or better. In some variations, the formulations and pharmaceutical formulations are administered periocularly, including without limitation subconjunctivally, to the eye of a huinan subject where the eye to which the formulation is administered has vision of 20/40 or better.

[0082] In some variations, the formulations and pharmaceutical formulations described herein are used to treat, prevent, or delay onset of AMD. In some variations, the formulations and phannaceutical formulations described herein are used to treat, prevent, or delay onset of dry AMD. In some variations, subjects including but not limited to huinan subjects with non-central geographic atrophy are administered a formulation or pharmaceutical formulations described herein to treat, prevent, or delay onset of central geographic atrophy. In some variations, the formulations and pharmaceutical formulations comprise a limus compound, including but not limited to rapamycin. In some variations the formulations and pharmaceutical fonnulations are administered periocularly, including without limitation subconjunctivally, to a huinan subject with vision of 20/40 or better. In some variations, the formulations and pharmaceutical formulations described herein are administered and the subject, including but not limited to a human subject is also treated with a second therapy for treating the disease or disorder. In some variations, the formulations and pharmaceutical formulations described herein are used to treat, prevent, or delay onset of wet or dry AMD and the subject, including but not limited to a human subject is also treated with laser therapy such as photodynamic laser therapy, either before, during, or after treatment with the fonnulations or pharmaceutical formulations described herein.

[0083] In some variations, the formulations and pharmaceutical formulations described herein are used to treat one or more of uveitis, allergic conjunctivitis, macular edema, glaucoma, or dry eye.

[0084] In some variations, a formulations or pharmaceutical formulation comprises a limus compound such as rapamycin, and is administered to treat, prevent, or delay onset of dry eye. In some variations, a formulations or pharmaceutical form.ulation comprises a limus compound such as rapamycin, and is administered to treat, prevent, or delay onset of allergic conjunctivitis.

[0085] In some variations, the formulations and pharmaceutical formulations described herein are used to treat glaucoma. In some variations, the formulations and pharmaceutical formulations described herein for treating glaucoma comprise a limus compound such as rapamycin, and are used as a surgical adjuvant to prevent, reduce or delay surgical complications. In some variations, the formulations and pharmaceutical formulations described herein for treating glaucoma comprise a limus compound such as rapamycin, and are used to improve or prolong surgical implant success. In some variations, the formulations and pharmaceutical formulations described herein for treating glaucoma comprise a limus compound such as rapamycin, and are used to improve or prolong success of an argon laser trabeculectomy or other glaucoma-related surgery. In some variations, the formulations and pharmaceutical formulations described herein have a neuroprotective effect and are used to treat glaucoma.

[00861 In some variations, the formtilations and pharmaceutical formulations described herein are used to treat retinitis pigmentosa. In some variations, the formulations and pharmaceutical formulations described herein for treating glaucoma comprise a limus compound such as rapamycin, and are used to treat, prevent, or delay onset of retinitis pigmentosa. In some variations, the formulations and phannaceutical formulations described herein have a neuroprotective effect and are used to treat retinitis pigmentosa.
[00871 hi some variations, the formulations and pharmaceutical formulations described herein are used to treat one or more of central retinal vein occlusive diseases (CRVO), branch retinal venous occlusion (BRVO), retinal vascular diseases and conditions, macular edema, diabetic macular edema, iris neovascularization, diabetic retinopathy, or corneal graft rejection. In some variations, a formulations or pharmaceutical formulation comprises a limus compound such as rapamycin, and is administered to treat, prevent, or delay onset of one or more of these diseases or conditions. In some variations the formulations and pharmaceutical formulations are administered subconjunctivally to an eye with vision of 20/40 or better.
[00881 When used to treat, prevent, inhibit, delay the onset of, or cause regressions of uveitis, the formulations and pharmaceutical formulations described herein may be administered by a variety of routes as is known in the art, including but not limited to by ocular or oral administration. Other routes of administration are lct-iown and are routine in the art. In some variations, the formulations described herein comprise rapamycin and are used to treat uveitis.
[00891 One disease that may be treated, prevented, inhibited, have onset delayed, or be caused to regress using the formulation, liquid formulations and methods described herein is the wet form of AMD. In some variations wet AMD is treated using the formulations, liquid formulations and methods described herein. The wet form of AMD is characterized by blood vessels growing from their normal location in the choroid into an undesirable position under the retina. Leakage and bleeding from these new blood vessels results in vision loss and possibly blindness.
__---[0090) The formulations, liquid formulations, and methods described herein may also be used to prevent or slow the transition from the dry form of AMD (wherein the retinal pigment epithelium or RPE degenerates and leads to photoreceptor cell death and the formation of yellow deposits called drusen under the retina) to the wet form of AMD.
[00911 "Macular degeneration" is characterized by the excessive buildup of fibrous deposits in the macula and retina and the atrophy of the retinal pigment epithelilun. As used hereiil, an eye "afflicted" with macular degeneration is understood to mean that the eye exhibits at least one detectable physical characteristic associated with the disease of macular degeneration. The administration of rapamycin appears to limit and regress angiogenesis, such as choroidal neovascularization in age-related macular degeneration (AMD), which may occur without treatment. As used herein, the term "angiogenesis"
means the generation of new blood vessels ("neovascularization") into a tissue or organ.
An "angiogenesis-mediated disease or condition" of the eye or retina is one in which new blood vessels are generated in a pathogenic manner in the eye or retina, resulting in dimunition or loss of vision or other problem, e.g., choroidal neovascularization associated with AMD.

[00921 The formulations and liquid formulations described herein, including but not limited to rapamycin-containing formulations and liquid formulations, may also be used to treat, prevent, inhibit, delay the onset of, or cause regression of various immune-related diseases and conditions, including but not limited to organ transplant rejection in a host, graft vs. host disease, autoimmune diseases, diseases of inflammation, hyperproliferative vascular disorders, solid tumors, and fungal infections. In some variations, the formulations and liquid formulations described herein, including but not limited to rapamycin-containing formulations and liquid formulations, are used to treat various immune-related diseases and conditions, including but not limited to organ transplant rejection in a host, graft vs. host disease, autoimmune diseases, diseases of inflammation, hyperproliferative vascular disorders, solid tumors, and fungal infections.
The formulations and liquid formulations described herein, including but not limited to rapamycin-containing formulations and liquid fonnulations, may be used as immunosuppressants. The formulations and liquid formulations described herein, including but not limited to rapamycin-containing formulations and liquid formulations, may be used to treat, prevent, inhibit, or delay the onset of rejection of transplanted organs or tissues including but not limited to transplanted heart, liver, kidney, spleen, lung, small bowel, pancreas, and bone marrow. In some variations, the formulations and liquid formulations described herein are used to treat the onset of rejection of transplanted organs or tissues including but not liinited to transplanted heart, liver, kidney, spleen, lung, small bowel, pancreas, and bone marrow. When used to treat, prevent, inhibit, delay the onset of, or cause regressions of immune-related diseases, including but not limited to transplant rejection, the formulations and liquid formulations described herein may be administered by a variety of routes as is known in the art, including but not limited to by oral administration.
(0093] Systemic administration may be achieved by oral administration of the liquid formulation. Other systemic routes of administration are known and are routine in the art.
Some examples thereof are listed in the Detailed Description section.

[0094] As used herein, to "inhibit" a disease or condition by administration of a therapeutic agent means that the progress of at least one detectable physical characteristic or symptom of the disease or condition is slowed or stopped following administration of the therapeutic agent as compared to the progress of the disease or condition without administration of the therapeutic agent.
[0095] As used herein, to "prevent" a disease or condition by administration of a therapeutic agent means that the detectable physical characteristics or symptom of the disease or condition do not develop following administration of the therapeutic agent.
[0096] As used herein, to "delay onset of' a disease or condition by administration of a therapeutic agent means that at least one detectable physical characteristic or symptom of the disease or condition develops later in time following adininistration of the therapeutic agent as compared to the progress of the disease or condition without administration of the therapeutic agent.
[0097] As used herein, to "treat" a disease or condition by administration of a therapeutic agent means that the progress of at least one detectable physical characteristic or symptom of the disease or condition is slowed, stopped, or reversed following administration of the therapeutic agent as compared to the progress of the disease or condition without administration of the therapeutic agent.

[0098] As used herein, to "cause regression of' a disease or condition by administration of a therapeutic agent means that the progress of at least one detectable physical characteristic or symptom of the disease or condition is reversed to some extent following administration of the therapeutic agent.
[0099] A subject, including but not limited to a human subject, having a predisposition for or in need of prevention may be identified by the skilled practitioner by established methods and criteria in the field given the teachings herein. The skilled practitioner may also readily diagnose individuals as in need of inhibition or treatment based upon established criteria in the field for identifying angiogenesis and/or neovascularization given the teachings herein.

[00100] As used herein, a"subject" is generally any animal that may benefit from administration of the therapeutic agents described herein. In some variations the therapeutic agents are administered to a mammalian subject. In some variations the therapeutic agents are administered to a human subject. In some variations the therapeutic agents may be administered to a veterinary animal subject. In some variations the therapeutic agents may be administered to a model experimental animal subject.

[0100] Other diseases and conditions that may be treated, prevented, inhibited, have the onset delayed, or be caused to regress using the methods described herein include those disclosed in the following patents and publications, the contents of each of which is incorporated herein in its entirety: PCT publication WO 2004/027027, published April 1, 2004, titled Method of inhibiting choroidal neovascularization, assigned to Trustees of the University of Pemisylvania; US patent number 5,387,589, issued February 7, 1995, titled Method of Treating Ocular Inflammation, with inventor Prassad Kulkarni, assigned to University of Louisville Research Foundation; US patent number 6,376,517, issued April 23, 2003, titled Pipecolic acid derivatives for vision and memory disorders, assigned to GPI NIL Holdings, Inc; PCT publication WO 2004/028477, published Apri18, 2004, titled Method subretinal adininistration of therapeutics including steroids:
method for localizing pharmadynamic action at the choroid and retina; and related methods for treatinent and or prevention of retinal diseases, assigned to hmorx, Inc; US
patent number 6,416,777, issued July 9, 2002, titled Ophthalmic drug delivery device, assigned to Alcon Universal Ltd; US patent number 6,713,081, issued March 30, 2004, titled Ocular therapeutic agent delivery device and methods for making and using such devices, assigned to Department of Health and Human Services; and U.S. Pat. No.
5,536,729, issued July 16, 1996, titled Rapamycin Formulations for Oral Administration, assigned to American Home Products Corp., and U.S. Pat. App. Nos. 60/503,840 and 10/945,682.
Liquid forfnulatious [0101] The liquid formulations described herein contain a therapeutic agent and may generally be any liquid formulation, including but not limited to solutions, suspensions, and emulsions. In some variations the liquid foi7nulations form a non-dispersed mass relative to a surrounding medium when placed in the vitreous of a rabbit eye.

[0102] When a certain volume of a liquid foimulation is administered, it is understood that there is some iinprecision in the accuracy of various devices that may be used to administer the liquid formulation. Where a certain volume is specified, it is understood that this is the target volume. However, certain devices such as insulin syringes are inaccurate to greater than 10%, and sometimes iilaccurate to greater than 20%
or more.
Hamilton HPLC type syringes are generally considered precise to within 10%, and are recommended for volumes below 10 l are to be injected.

[0103] In some variations, a volume of a liquid formulation described herein is adininistered to the vitreous of a rabbit eye or a subject's, including but not limiting a human subject's eye that is less than about 500 l, less than about 400 l, less than about 300 l, less than about 200 ,ul, less than about 100 ,ul, less than about 90 l, less than about 80 ,ul, less than about 70 l, less than about 60 ,ul, less than about 50 l, less than about 40 ,ul, less than about 30 l, less than about 20 l, less than about 10 ,ul, less than about 5 l, less than about 3 l, or less than about 1 l. In some variations, a volume of a liquid formulation described herein is administered to the vitreous of a rabbit eye or subject's, including but not limited to a human subject's eye that is less than about 20 l.
In some variations, a volume of a liquid formulation described herein is administered to the vitreous that is less than about 10 l. In some variations, a volume of a liquid formulation described herein is administered to the vitreous of a rabbit eye or a subject's, including but not limited to a human subject's eye that is between about 0.l l and about 200 l, between about 50 l and about 200 l, between about 50 l and about 150 l, between about 0.l l and about 100 l, between about 0.1 1 and about 50,u1, between about l l and about 40 l, between about l l and about 30 l, between about 1 l and about 20 l, between about l l and about 10 l, or between about l l and about 5 l. In some variations, a volume of a liquid formulation described herein is administered to the vitreous of a rabbit eye or a subject's, including but not limited to a human subject's eye that is between about 1 1 and about 10 l. In some variations, a volume of a liquid formulation described herein is administered to the vitreous of a rabbit eye or a subject's, including but not limited to a human subject's eye that is between about l l and about 5 _ -----l. In some variations, a volume of a liquid formulation described herein is administered to the vitreous of a rabbit eye or a subject's eye that is between about 1 l and about 5 l.

In some variations, a volume of a liquid formulation described herein is administered to the vitreous of a rabbit eye or a subject's, including but not limited to a human subject's eye that is between about 0.1 l and about 200 l.
[0104] In some variations, a total voh.ime of a liquid formulation described herein is subconj unctivally administered to a rabbit eye or a subj ect's, including but not limited to a human subject's eye that is less than about 1000 l, less than about 900 l, less than about 800 1, less than about 700 ,u,l, less than about 600 l, less than about 500 l, less than about 400 l, less than about 300 l, less than about 200 l, less than about 100 l, less than about 90 1, less than about 80 l, less than about 70 1, less than about 60 ,ul, less than about 50 l, less than about 40 l, less than about 30 l, less than about 20 l, less than about 10 l, less than about 5 l, less than about 3 l, or less than about 1 l. In some variations, a voh.ime of a liquid formulation described herein is subconjunctivally administered to a rabbit eye or a subject's, including but not limited to a huinan subject's eye that is less than about 20 l. In some variations, a volume of a liquid formulation described herein is subconjunctivally administered to a rabbit eye or a subject's, including but not limited to a human subject's eye that is less than about 10 l. In some variations, a volume of a liquid formulation described herein is subconjunctivally administered to a rabbit eye or a subject's, including but not limited to a humaii subject's eye that is between about O.l l and about 200 l, between about 50 l and about 200 l, between about 200 I and about 300 1, between about 300 1 and about 400 l, between about 400 l and about 500 l, between about 600 l and about 700 l, between about 700 l and about 800 l, between about 800 l and about 900 l, between about 900 l and about 1000 ft1, between about 50 l and about 150 l, between about 0.l l and about 100 l, between about 0.1 1 and about 50 1, between about l l and about 40 1, between about l l and about 30 1, between about l l and about 20 l, between about l l and about 10 l, or between about l l and about 5 1. In some variations, a volume of a liquid formulation described herein is subconjunctivally administered to a rabbit eye or a subject's, including but not limited to a human subject's eye that is between about 1 1 and about 10 l. In some variations, a volume of a liquid formulation described herein is subconjunctivally administered to a rabbit eye or a subject's, including but not limited to a human subject's eye that is between about l l and about 5 l. In some variations, a volume of a liquid formulation described herein is administered to subconjunctivally administered to a rabbit eye or a subject's, including but not limited to a human subject's eye that is between about l i and about 5 l. In some variations, a volume of a liquid formulation described herein is administered to subconjunctivally administered to a rabbit eye or a subject's, including but not limited to a human subject's eye that is between about 0.1 1 and about 200 l.
[0105] In some variations the liquid formulations described herein are administered in multiple subconjunctival locations within a period of time, including without limitation witllin an hour of one another. Without being bound by theory, it is thought that such multiple administrations, such as multiple injections, allow for a greater total dose to be administered subconjunctivally than a single dose due to a potentially limited ability of the local ocular tissues to absorb larger volumes.
[0106] One liquid formulation described herein is an in situ gelling formulation. In situ gelling formulations, as described herein, coinprise a therapeutic agent and a plurality of polymers which give a formulation that forms a gel or a gel-like substance when placed in an aqueous medium, including but not limited to an aqueous medium of the eye.
[0107] In some variations of the liquid formulations described herein, the therapeutic agent is a solution or suspension of rapamycin in a liquid medium. Liquid media include but are not limited to solvents, including but not limited to those in the Solubilizatiot2 of Tlzerapeutic Agents section.
[0108] The liquid formulations described herein may comprise a solubilizing agent component. In some variations the solubilizing agent component is a surfactant. Note that there is some overlap between components that may be solvents and solubilizing agents, and therefore the same component may in some systems be used as either a solvent or a solubilizing agent. A liquid formulation that comprises a therapeutic agent and a component that may be considered either a solvent or a solubilizing agent or surfactant will be considered a solvent if it is playing the role of a solvent; if the conlponent is not playing the role of the solvent, the component may be considered a solubilizing agent or surfactant.
[0109] Liquid formulations may optionally further comprise stabilizers, excipients, gelling agents, adjuvants, antioxidants, and/or other components as described herein.
[0110] In some variations all coinponents in the liquid formulation, other than the therapeutic agent, are liquid at room temperature.
_ ----- - --- --- -[0111] In some variations, the liquid formulation comprises a release modifying agent.
Tn some variations, the release modifying agent is a film-forming polymer component.

The film-forming polyiner component may comprise one or more film-forming polymers.
Any film-forming polymer may be used in the excipient component. In some variations, the film-forming polymer component comprises a water insoluble film fonning polymer.
In some variations, the release modifying agent component comprises an acrylic polymer, including but not limited to polymethacrylate, including but not limited to Eudragit RL.
[0112] Described herein are compositions and liquid forinulations for delivery of the therapeutic agents described in the Tlaerapeutic Agents section. Delivery of therapeutic agents using the compositions and liquid fornnulations described herein may be used to treat, prevent, inhibit, delay the onset of, or cause the regression of the diseases and conditions described in the Diseases and Conditions section. The compositions and liquid formulations described herein may comprise any of the therapeutic agents described in the Tlzerapeutic Agents section, including but not limited to rapamycin.
The compositions and liquid formulations described herein may comprise one or more than one therapeutic agent. Other compositions and liquid formulations in addition to those explicitly described herein may be used.

[0113] When the therapeutic agent is rapamycin, the compositions and liquid formulations may be used to maintain an amount of rapamycin in the vitreous effective to treat wet AMD. In one nonlimiting example, it is believed that a liquid formulation delivering rapamycin to maintain a concentration of rapamycin of about 10 pg/ml to about 2 g/ml in the vitreous over a period of time may be used for the treatment of wet AMD.
When the rapamycin is in a liquid formulation that forms a non-dispersed mass, the stated concentration of rapamycin represents the amount that is effectively treating the disease or condition of the eye, and not merely present in the form of the non-dispersed mass. In another nonlimiting example, it is believed that a delivery system delivering rapamycin to maintain a concentration of rapamycin of about 0.01 pg/mg to about 10 ng/mg in the retina choroid tissues over a period of time may be used for treatment of wet AMD. Other therapeutically effective amounts of therapeutic agent are also possible, and can be readily determined by one of skill in the art given the teachings herein.

[0114] When the therapeutic agent is rapamycin, the compositions and liquid formulations described herein may be used to deliver a dose of rapamycin to a subject, including but not limited to a human subject or to the eye of a subject. In one nonlimiting example, it is believed that a liquid formulation containing a dose of about 20 g to about 4 mg may be used for the treatment of wet AMD.

[0115] In some variations the therapeutic agent in the liquid formulation comprises between about 0.01 to about 30% of the total weight of the composition;
between about 0.05 to about 15%; between about 0.1 to about 10%; between abottt 1 to about 5%; or between about 5 to about 15%; between about 8 to about 10%; between about 0.01 to about 1%; between abotit 0.05 to about 5%; between abottt 0.1 to about 0.2%;
between about 0.2 to about 0.3%; between about 0.3 to about 0.4%; between about 0.4 to about 0.5%; between about 0.5 to about 0.6%; between about 0.6 to about 0.7%;
between about 0.7 to about 1%; between about 1 to about 5%; between about 5 to about 10%;
between about 15 to about 30%, between about 20 to about 30%; or between about 25 to about 30%.

[0116] Those of skill in the art, based on the teachings herein can determine what amount or concentration of a given therapeutic agent is equivalent to an amount or concentration of rapamycin by, for example, administering the therapeutic agent at various amounts or concentrations to a disease model system, such as an in vivo or in vivo model system, and comparing the results in the model system relative to the results of various amounts or concentrations of rapamycin. Those of skill in the art, based on the teachings herein can also determine what amount or concentration of a given therapeutic agent is equivalent to an amount or concentration of rapamycin by reviewing the scientific literature for experiments performed comparing rapainycin to other therapeutic agents. It is understood that even the same therapeutic agent may have a different equivalent level of rapamycin when, for example, a different disease or disorder is being evaluated, or a different type of formulation is used. Nonlimiting examples of scientific references with comparative studies of rapamycin and other therapeutic agents on ocular disease are Ohia et al., Effects of steroids and immunosuppressive drugs on endotoxin-uveitis in rabbits, J.
Ocul. Pharmacol. 8(4):295-307 (1992); Kulkami, Steroidal and nonsteroidal drugs in endotoxin-induced uveitis, J. Ocul. Pharmacol. 10(1):329-34 (1994); Hafizi et al., Diffet=ential effects of rapamycin, cyclosporine A, and FK506 on human coronary artery snaooth nauscle cell proliferation and signaling, Vascul Pharmacol. 41(4-5):167-76 (2004); and US 2005/0187241.

[0117] For example, in a model for wet AMD, if a therapeutic agent is found to be approximately 10-fold less potent or efficacious than rapamycin in the treatment of wet AMD, a concentration of 10 ng/ml of the therapeutic agent would be equivalent to a 1 ng/ml concentration of rapamycin. Or if a therapeutic agent is found to be approximately 10-fold less potent or efficacious than rapainycin in the treatment of wet AMD, a 10-fold amount of the therapeutic agent would be administered relative to the amount of rapamycin.

[0118] The solvent component may comprise, for instance, between about 0.01 to about 99.9 % of the total weight of the composition; between about 0.1 to abotit 99%; between about 25 to about 55%; between about 30 to about 50%; or between about 35 to about 45%; between about 0.1 to about 10%; between about 10 to about 20%; between about 20 to about 30%; between about 30 to about 40%; between about 40 to about 45%;
between about 40 to abotit 45%; between about 45 to about 50%; between about 50 to about 60%;
between about 50 to about 70%; between about 70 to about 80%; between about 80 to about 90%; or between about 90 to about 100%.

[0119] The solubilizing agent component may comprise, for instance, between about 0.01 to about 30 % of the total weight of the composition; between about 0.1 to about 20%; between about 2.5 to about 15%; between about 10 to about 15 %; or between about to about 10%; between about 8 to about 12%; between about 10 to about 20%;
between about 20 to about 30%.

[0120] In some variations, the liquid formulations described herein have a viscosity of between 40% and 120% centipoise. In some variations the liquid formulations described herein have a viscosity of between 60% and 80% centipoise.

[0121] In some variations the liquid formtilations described herein comprise a therapeutic agent and a solvent component. The solvent component may comprise a single solvent or a combination of solvents. The therapeutic agent component may comprise a single therapeutic agent or a combination of therapeutic agents. In some variations, the solvent is glycerin, dimethylsulfoxide, N-methylpyrrolidone, dimethyl acetamide (DMA), dimethyl formamide, glycerol formal, ethoxy diglycol, triethylene glycol dimethyl ether, triacetin, diacetin, corn oil, acetyl triethyl citrate (ATC), ethyl lactate, polyglycolated capryl glyceride, Tbutyrolactone, dimethyl isosorbide, benzyl alcohol, ethanol, isopropyl alcohol, polyethylene glycol of various molecular weights, including but not limited to PEG 300 and PEG 400, or propylene glycol, or a mixture of one or more thereof.

[0122] In some variations the liquid fonnulations described herein are solutions, and comprise a therapeutic agent and a solvent component. In some variations the solvent component comprises ethanol. In some variations the solvent component comprises ethanol and a polyethylene glycol, including but not limited to a liquid polyethylene glycol, including but not limited to one or more of PEG 300 or PEG 400.

[0123] In some variations the liquid formulations described herein contain no greater than about 250 ttl of polyethylene glycol. In some variations the liquid formulations described herein contain no greater than about 250 l, no greater than about 200 l, no greater than about 150 [Cl, no greater than about 125 l, no greater than about 100 l, no greater than about 75 l, no greater than about 50 l, no greater than about 25 l, no greater than about 20 l, no greater than about 15 l, no greater than about 10 l, no greater than about 7.5 l, no greater than about 5 l, no greater than about 2.5 l, no greater than about 1.0 .l, or no greater than about 0.5 l of polyethylene glycol.
Formulations containing polyethylene glycol may contain, for example, PEG 300 or PEG
400.
[0124] In some variations, the liquid formulations described herein are suspensions, and comprise a therapeutic agent and a diluent component. In some variations, the diluent component comprises one or more components listed herein as solvents or solubilizing agents, wherein the resulting mixture is a suspension.
[0125] In some variations the liquid formulation is partly a solution and partly a suspension.
[0126] In some variations the liquid formulation is an in situ gelling formulation, and comprises a therapeutic agent and a polyiner component, wherein the polymer component may comprise a plurality of polymers. In some variations, the liquid formulation comprises a polymethacrylate polymer. In some variations, the liquid formulation comprises a polyvinylpyrrolidone polymer.
[0127] Some variations of liquid formulations include a therapeutic agent or agents such as but not limited to rapamycin between about 0.01 % and about 20% by weight of the total, a solvent between about 5% and about 15% by weight of the total, a solubilizing agent including but not limited to a surfactant between about 5% and about 15%
by weight of the total, with water as the primary remaining component. In some variations the formulations fttrther comprise stabilizing agents, excipients, adjuvants, or antioxidants, between about 0 and about 40% by weight of the total.

[0128] In some variations, a liquid formulation comprises up to about 5%
therapeutic -- - - ---agent, including but not limited to rapamycin, per weight of the total; and up to about 99.9% of a solvent component, by weight of the total. In some variations the liquid formulation coinprises up to about 5% therapeutic agent, including but not limited to rapamycin, per weight of the total; and up to about 99.9% of a diluent component.

[0129] In some variations, a liquid formulation may comprise up to about 5%
therapeutic agent, including but not limited to rapamycin, per weight of the total; up to about 10% solvent by weight of the total; and up to about 85% of a solubilizing component, by weight of the total. In some variations the solubilizing component is an aqueous solution of a surfactant.
[0130] A plurality of polymers coinponent may comprise, for instance, between about 0.01 to about 30 % of the total weight of the composition; between about 0.1 to about 20%; between about 2.5 to about 15%; between about 10 to about 15%; between about 3 to about 5%; between about 5 to about 10%; between about 8 to about 12%;
between about 10 to about 20%; or between about 20 to about 30%.
[0131] Some variations of liquid formulations includes a therapeutic agent or agents such as but not limited to rapamycin between about 0.01% a.nd about 20% by weight of the total, a solvent component between about 60% and about 98% by weight of the total, and a plurality of polymers, whose combined percentage is between about 0.1 %
and about 15% by weight of the total. In some variations the formulations further comprise stabilizing agents, excipients, adjuvants, or antioxidants, between about 0 and about 40%
by weight of the total.
[0132] In some variations, a liquid fonnulation may comprise about 4%
therapeutic agent, including but not limited to rapamycin, per weight of the total; about 91% solvent by weight of the total; and about 5% polymeric coiuponent, per weight of the total.
[0133] Some examples and variations of liquid formulations described herein were prepared and are listed in Table 1. Depending on their type, the listed formulations are denoted one or more of solutions ("S"), suspensions ("SP"), emulsions ("E") or in situ gelling ("ISG"). Median particle size is listed for some of the suspensions.
As described herein, some liquid formulations form a non-dispersed mass after, for example, injection into an aqueous environment such as the vitreous of an eye. For those formulations injected into the vitreous of a rabbit eye, the right-hand column of Table 1 indicates whether or not a non-dispersed mass (NDM) formed after a specified volume was injected into the vitreous of the rabbit eye.
[0134] The following references, each of which is incorporated herein by reference in its entirety, show one or more formulations, including but not limited to rapamycin formulations, and which describe use of rapamycin at various doses and other therapeutic ageilts for treating various diseases or conditions: US 60/651,790, filed 2/9/2005, titled FORMULATIONS FOR OCULAR TREATMENT, attorney docket number 57796-30002.00; US 60/664,040, filed 2/9/2005, attorney docket number 57796-30004.00, titled LIQUID FORMULATIONS FOR TREATMENT OF DISEASES OR CONDITIONS; US
60/664,119, filed 3/21/2005, attorney docket number 57796-30005.00, titled DRUG
DELIVERY SYSTEMS FOR TREATMENT OF DISEASES OR CONDITIONS; US
60/664,306, filed 3/21/2005, attorney docket number 57796-30006.00 titled IN
SITU
GELLING FORMULATIONS AND LIQUID FORMULATIONS FOR TREATMENT
OF DISEASES OR CONDITIONS; US ,/ , filed 2/9/2006, titled FORMULATIONS
FOR OCULAR TREATMENT, attorney docket number 57796-20002.00; /_, filed 2/9/2006, attorney docket number 57796-20004.00, titled LIQUID FORMULATIONS
FOR TREATMENT OF DISEASES OR CONDITIONS; US 2005/0187241, and US
2005/0064010.

Liquid forszzulatiou.s wlziclz fornz a non-dispersed nzass [0135] One class of liquid formulations described herein forms a non-dispersed mass when placed in an aqueous medium. As used herein, a "non-dispersed mass"
refers to the structure formed or shape assumed when the liquid formulation is placed into an environment, relative to the environment in which it is placed. Generally, a non-dispersed mass of a liquid formulation is anything other than a homogeneous distribution of the liquid formulation in the surrounding medium. The non-dispersed mass may, for instance, be indicated by visually inspecting the administered liquid formulation and characterizing its appearance relative to the surrounding medium.
[01361 In some variations, the aqueous medium is water. In some variations, the water is deionized, distilled, sterile, or tap water, including but not limited to tap water available at the place of business of MacuSight in Union City, California.

[0137] In some variations, the aqueous medium is an aqueous mediuiu of a subject. In some variations the aqueous medium is an aqueous medium of the eye of a subject, including but not limited to the vitreous of an eye of a subject. In some variations the subject is a human subject. In some variations the subject is a rabbit. _ [0138] In some variations the liquid formulation forms a non-dispersed mass when exposed to a certain temperature or range of temperatures, including but not limited to about room temperature, about ambient temperature, about 30 C, about 37 C, or about the temperature of the aqueous medium of the subject.
[0139] In some variations the liquid fornnulation forms a non-dispersed mass when exposed to a certain pH or range of pH, including but not liinited to a pH
between about 6 and about 8.
[0140] In some variations, the non-dispersed mass comprises a gel or gel-like substance.
[0141] In some variations, the non-dispersed mass comprises a polymer niatrix.
In some variations, the non-dispersed mass comprises a polymer matrix in which a therapeutic agent is dispersed.
[0142] The liquid formulations described lierein may generally be of any geometry or shape after administration to a subject or the eye of a subject, including but not limited to a human subject. In some variations, the non-dispersed mass is between about 0.1 and about 5 mm. In some variations, the non-dispersed mass is between about 1 and about 3 mm. The non-dispersed mass-forming liquid formulations may, for instance, appear as a compact spherical mass when administered to the vitreous. In some -instances, the liquid fonnulation may appear as a non-dispersed mass relative to the surrounding medium, wherein the non-dispersed mass is less clearly defined and the geometry is more amorphous than spherical.
[0143] The non-dispersed mass-forming liquid fonnulations described herein may form a non-dispersed mass immediately upon placement in the medium or the non-dispersed mass may form some period of time after placement of the liquid formulation.
In some variations the non-dispersed mass forms over the course of about 1, about 2, about 3, about 4, about 5, about 6, or about 7 days. In some variations the non-dispersed mass forms over the course of about 1 week, about 2 weeks, or about 3 weeks.

[0144) In some variations, the liquid formulations described herein that fonn a non-dispersed mass appear as a milky or whitish colored semi-contiguous or semi-solid non-dispersed mass relative to the medium in which it is placed.
(0145] One liquid formulation described herein forms a non-dispersed mass which has the form of a solid depot when the formulation is injected into any or all of water, the vitreous of a rabbit eye, or between the sclera and the conjunctiva of a rabbit eye. One liquid formulation described herein forms a non-dispersed mass which has the form of a semi-solid when the formulation is injected into any or all of water, the vitreous of a rabbit eye, or between the sclera and the conjunctiva of a rabbit eye. One liquid formulation described herein forms a non-dispersed mass which has the form of a polyineric matrix when the formulation is injected into any or all of water, the vitreous of a rabbit eye, or between the sclera and the conjunctiva of a rabbit eye. One liquid formulation described herein forms a non-dispersed mass wliich has the form of a gel, a hydrogel, or a gel-like substance when the formulation is injected into any or all of water, the vitreous of a rabbit eye, or between the sclera and the conjunctiva of a rabbit eye.
[0146] In some variations described herein the liquid formulatioi-i forms a non-dispersed mass relative to a surrounding medium where the surrounding medium is aqueous.
An "aqueous medium" or "aqueous environment" is one that contains at least about 50%
water. Exainples of aqueous media include but are not limited to water, the vitreous, extracellular fluid, conjunctiva, sclera, between the sclera and the conjunctiva, aqueous humor, gastric fluid, and any tissue or body fluid coinprised of at least about 50% of water. Aqueous media include but are not limited to gel structures, including but not limited to those of the conjunctiva and sclera.

[0147] In some variations, the liquid formulations described herein form a non-dispersed mass when a test volume of the liquid formulation is placed in the vitreous of a rabbit eye. In some variations the test volume administered to a rabbit eye, and the test volume is equal to the volume of the liquid formulation administered to a subject's, including but not limited to a human subject's eye.

[0148] In some variations, the test volume administered to a rabbit eye is equal to the volume administered to the subject's eye multiplied by a scale factor, and the scale factor is equal to the average volume of a rabbit eye divided by the average volume of a subject eye. The "average volume" of an eye, as used herein, refers to the average volume of an eye of a member of similar age of the species under consideration generally, as opposed to the average volume of any particular individual's eye.

[0149] hi some variations, the test volume administered to the rabbit eye is between about 10 Al and about 50 l. In some variations, the test volume administered to the rabbit eye is between about 1 l and about 30 l. In some variations, the test volume administered to the rabbit eye is between about 50 l and about 100 .l. In some variations, the test volume administered to the rabbit eye is between about 25 Al and about 75 l. In some variations, the test volume administered to the rabbit eye is about 30 l.

[0150] In some variations, the liquid foimulation that forms a non-dispersed mass when placed in the inedium may comprises a therapeutic agent or agents with a concentration of between about 0.01 % and about 10% by weight of the total, and a solvent between about % and about 99 % by weight of the total. In some variations the formulation fiirther comprises a solubilizing agent including but not limited to a surfactant. In some variations the liquid formulation further comprises a stabilizing agent, excipient, adjuvant, or antioxidant, etc., between about 0 and abotit 40% by weight of the total.
In some variations, the therapeutic agent is about 5% by weight of the total, and the solvent component is about 95% by weight of the total.

[0151] Whether a liquid formulation exhibits a non-dispersed mass relative to a surrounding medium when present in a subject, including but not limited to a human subject or the eye of a subject may be determined by, for instance, mixing a therapeutic agent with a solvent, administering it to the vitreous of an eye of a subject, including but not limited to a human subject, and comparing the liquid formulation to the surrounding medium.

[0152] One liquid formulation that may be used for treating, preventing, inhibiting, delaying the onset of, or causing the regression of the diseases and conditions of a subject, including but not limited to a human subject, is a liquid formulation that forms a non-dispersed mass when placed into the vitreous of a rabbit eye. When used for treating, preventing, inhibiting, delaying the onset of, or causing the regression of the disease or condition of the subject, the liquid formulation is administered to the subject. The liquid formulation may or may not form a non-dispersed mass in the subject. One liquid formulation described herein forms a non-dispersed mass when administered to a subject and forms a non-dispersed mass when administered to a rabbit eye.

[0153] Without being bound by theory, it is believed that the low soltibility of rapamycin in the vitreous contributes to the formation of a non-dispersed mass by some rapamycin-containing liquid formulations described herein. The vitreous is a clear gel composed almost entirely of water (up to 99%). Without being bound by theory, it is believed that as rapamycin in an injected formulation contacts the vitreous, the rapamycin precipitates.

[0154] Without being bound by theory, factors believed to affect the formation of and geometry of a non-dispersed mass include the concentration of rapamycin in the formulation, the viscosity of the formulation, ethanol content of the formulation, and the volume of injection. It is believed that maintaining a higher local concentration of rapamycin after injection of the formulation favors formation of a non-dispersed mass, as opposed to a lower local concentration of rapamycin after injection of the forinulation. As volume is increased for a given dose, foimation of a non-dispersed mass may become less favorable. Formation of a non-dispersed mass may become more favorable as rapamycin concentration is increased and/or as viscosity is increased. Ethanol content affects both the solubility of the rapamycin in the formulation and the viscosity of the formulation.
[0155] In one comparison, 100 EL1 of a solution of 0.4 % rapamycin, 4.0%
ethanol, and 95.6% PEG 400 (a 400 g dose) did not forxn a non-dispersed mass after injection into a rabbit eye. In contrast, 20 l of a solution of 2.00 % rapamycin, 4.0%
ethanol, and 94%
PEG 400 (also a 400 g dose) formed a compact spherical non-dispersed mass after injection into a rabbit eye.

[0156] Without being bound by theory, in the latter example, it is hypothesized that formation of the non-dispersed mass occurred as depicted in Figures lA-1C and described as follows. Upon injection, due to its viscosity the liquid formulation formed a spherical globule 100 within the vitreous 110. Ethanol then diffused out of this globule, resulting in localized precipitation 120 of the rapamycin within the globule. Eventually, the polyetllylene glycol also diffused out of the globule to leave a solid, compact non-dispersed mass of rapamycin 130.

[0157] In some variations, the non-dispersed masses described herein consists of at least about 20%, at least about 30%, at least about 40%, at least about 50%, at least about 60%, at least about 70%, at least about 80%, at least about 90%, or at least about 95% by volume of therapeutic agent when injected into the vitreous of a rabbit eye.

[0158] In some variations, upon formation a non-dispersed mass comprising rapamycin, for example, delivers the drug continuously at approximately a constant rate for an extended period of time. Without being bound by theory, it is believed that delivery of rapamycin from a non-dispersed mass in the vitreous depends on dissolution of the rapamycin in the vitreous, which depends in turn on clearance of the drug from the vitreous to other tissues. Without being bound by theory, this release process is believed to maintain a steady-state concentration of rapamycin in the vitreous.

[0159] In some variations, formation of a non-dispersed mass_reduces the toxicity of the injected liquid formulation compared to an equivalent dose that did not form a non-dispersed mass. In variations in which a liquid formulation injected into the vitreous does not form a non-dispersed mass, the drug (e.g., rapamycin) appears to disperse in the vitreous body. In some variations this may interfere with vision.

[0160] In some variations, liquid formulations that are suspensions form a non-dispersed mass upon injection into the vitreous. Formation of a non-dispersed mass from an injected suspension may become more favorable as the suspension particle size increases.

[0161] In some variations, it is believed that the liquid formulations will form a visually observable non-dispersed mass when injected into the eye of a subject, including but not limited to a human subject.

[0162] In some variations, liquid formulations are believed to form non-dispersed masses when injected subconjunctivally. In some variations it is believed that when subconjunctivally administered the liquid formulation forms a depot in the scleral tissue.
That is, it is believed that the therapeutic agent is absorbed into the sclera proximate to the injection site and forms a local concentration of drug in the selera.

In situ Gelliizg Fornaulatious [0163] Described herein are non-dispersed mass-forming liquid formulations which form a gel or gel-like substance wlien placed in an aqueous medium. In some variations, the non-dispersed mass comprises a gel; in some variations the gel is a hydrogel.

[0164] An "in situ gelling formulation," as used herein, refers to a liquid formulation which forins a gel-like non-dispersed mass when the liquid formulation is placed in an aqueous medium, including but not limited to aqueous media that are water, the vitreous of a rabbit eye, and between the sclera and the conjunctiva of a rabbit eye.
In some variations, an in situ gelling formulation forms a gel-like non-dispersed mass when placed in tap water.

[0165] In some variations, the in situ gelling formulation is a suspension prior to placement in an aqueous medium, and forms a gel in situ upon placement in an aqueous medium. In some variations, the in situ gelling formulation is a solution prior to placement in an aqueous medium, and forms a gel in situ upon placement in an aqueous medium. In some variations, the in situ gelling formulation is an emulsion prior to placement in an aqueous medium, and forms a gel in situ upon placement in an aqueous mediuin. In some variations a gel-like non-dispersed mass forms after placement of the in situ gelling formulation into an aqueous medium, including but not limited to any or all of water, the vitreous, or between the sclera and the conjunctiva of an eye. In some variations, the in situ gel is formed of a polyiner matrix. In some variations a therapeutic agent is dispersed in the polyrner matrix.

[0166] Described herein are in situ gelling formulations which may be used for treating, preventing, inhibiting, delaying the onset of, or causing the regression of the diseases and conditions of a subject including but not limited to a human subject. When used for treating, preventing, inhibiting, delaying the onset of, or causing the regression of the disease or condition of the subject, the in situ gelling formulation is administered to the subject. One liquid formulation described herein comprises an in situ gelling formulation which forms a non-dispersed mass when administered to a subject and forms a non-dispersed mass when administered to a rabbit eye.

[0167] In some variations, the in situ gelling formulation comprises one or more polymers. Described herein are various types of polymers, including polymers which are solvents, polymers which are solubilizing agents, polymers which are release modifying agents, polymers which are stabilizing agents, etc. In some variations, any combination of polymers is used wherein the polymers when combined with the therapeutic agent form any or all of a non-dispersed mass, a gel, a hydrogel, or polymeric matrix when placed in an aqueous medium, including but not limited to any or all of water, the vitreous, or between the sclera and the conjunctiva.

[0168] In some variations, the in situ gelling formulation delivers extended release of therapeutic agents to a subject when administered to the subject.
[0169] In some variations, the liquid formulation comprises a therapeutic agent and a plurality of polymers, wherein one of the polymers is a polymethacrylate.
Polymethacrylates are known by various names and are available in various preparations, including but not limited to polyineric methacrylates, methacrylic acid-ethyl acrylate copolymer (1:1), methacrylic acid-ethyl acrylate copolymer (1:1) dispersion 30 per cent, methacrylic acid-methyl methacrylate copolymer (1:1), methacrylic acid-metliyl methacrylate copolymer (1:2), acidum methacrylicum et ethylis acrylas polymerisatunl 1:1, acidum methacrylicum et ethylis acrylas polymerisatum 1:1 dispersio 30 per centum, acidum methacrylicum et methylis methacrylas polymerisatum 1:1, acidum methacrylicum et methylis methacrylas polymerisatum 1:2, USPNF: ammonio inethacrylate copolymer, methacrylic acid copolymer, methacrylic acid copolymer dispersion.
[0170] In some variations, one of the polymers is polyvinylpyrrolidone.
Polyvinylpyrrolidone is known by various names and is available in various preparations, including but not limited to povidone, povidonum, kollidon; plasdone; poly[1-(2-oxo-1-pyrrolidinyl)ethylene]; polyvidone; PVP; 1-vinyl-2- pyrrolidinone polymer, and Ethenyl-2-pyrrolidinone homopolymer.
[01711 One liquid formulation described herein comprises a therapeutic agent and a solvent coinponent. The solvent component may comprise a single solvent or a combination of solvents.
[01721 In some variations, the solvent is glycerin, dimethylsulfoxide, N-methylpyrrolidone, ethanol, isopropyl alcohol, polyethylene glycol of various molecular weights, including but not limited to PEG 300 and PEG 400, or propylene glycol, or a mixture of one or more thereof.
[0173] In some variations, the solvent is polyethylene glycol. Polyethylene glycol is known by various names and is available in various preparations, including but not limited to macrogels, macrogel 400, macrogel 1500, macroge14000, macrogel 6000, macrogel 20000, macrogola, breox PEG; carbowax; carbowax sentry; Hodag PEG;
Lipo;
Lipoxol; Lutrol E; PEG; Pluriol E; polyoxyethylene glycol, and cx Hydro-w-hydroxy-poly(oxy-1,2-ethanediyl) .

C'otnpositions and liquid forinulations for delivery of tlzerapeutic agents [0174] The compositions and liquid formulations described herein may be used to deliver amounts of the therapeutic agents effective for treating, preventing, inhibiting, delaying on set of, or causing the regression of the diseases and conditions described in the Diseases and Conditions section. In some variations the compositions and liquid formulations described herein deliver one or more therapeutic agents over an extended period of time.
[0175] An "effective amount," which is also referred to herein as a "therapeutically effective amount," of a therapeutic agent for administration as described herein is that amount of the therapeutic agent that provides the therapeutic effect sought when administered to the subject, including but not limited to a human subject. The achieving of different therapeutic effects may require different effective amounts of therapeutic agent. For example, the therapeutically effective amount of a therapeutic agent used for preventing a disease or condition may be different from the therapeutically effective amount used for treating, ii-iliibiting, delaying the onset of, or causing the regression of the disease or condition. h-i addition, the therapeutically effective amount may depend on the age, weight, and other health conditions of the subject as is well know to those versed in the disease or condition being addressed. Thus, the therapeutically effective amount may not be the saine in every subject to which the therapeutic agent is administered.

[0176] An effective ainount of a therapeutic agent for treating, preventing, inhibiting, delaying the onset of, or causing the regression of a specific disease or condition is also referred to herein as the amount of therapeutic agent effective to treat, prevent, inhibit, delay the onset of, or cause the regression of the disease or condition.

[0177] To determine whether a level of therapeutic agent is a "therapeutically effective amount" to treat, prevent, inhibit, delay on set of, or cause the regression of the diseases and conditions described in the Diseases and Coixditioyxs section, liquid formulations may be administered in animal models for the diseases or conditions of interest, and the effects may be observed. In addition, dose ranging huinan clinical trials may be conducted to determine the therapeutically effective amount of a therapeutic agent.

[0178] Generally, the therapeutic agent may be formulated in any composition or liquid formulation capable of delivery of a therapeutically effective amount of the therapeutic agent to a subject or to the eye of a subject for the required delivery period. Compositions include liquid formulations.

Solubilizatiofz of therapeutic ageuts [0179] One composition or liquid formulation that may be used is a composition or liquid formulation in which the therapeutic agent is dissolved in a solvent component.
Generally, any solvent which has the desired effect may be used in which the therapeutic agent dissolves. In some variations the solvent is aqueous. In some variations the solvent is non-aqueous. An "aqueous solvent" is a solvent that contains at least about 50% water.
[0180] Generally, any concentration of solubilized therapeutic agent that has the desired effect can be used. The solvent component may be a single solvent or may be a mixture of solvents. The solvent component may be a single solvent or may be a mixture of solvents. Solvents and types of solutions are well known to those versed in such drug delivery technologies. See for example, Remington: The Science and Practice of Pharmacy, Twentieth Edition, Lippincott Williams & Wilkins; 20th edition (December 15, 2000); Ansel's Pharmaceutical Dosage Forms and Dnig Delivery Systems, Eighth Edition, Lippincott Williams & Wilkins (August 2004); Handbook Of Pharmaceutical Excipients 2003, American Pharmaceutical Association, Washington, DC, USA and Pharmaceutical Press, London, UK; and Strickley, solubilizing Excipients in Oral and Injectable Fomiulations, Pharmaceutical Research, Vol. 21, No. 2, February 2004.
[0181) As noted previously, some solvents may also serve as solubilizing agents.
[0182] Solvents that may be used include but are not limited to DMSO, ethanol, methanol, isopropyl alcohol; castor oil, propylene glycol, glycerin, polysorbate 80, benzyl alcohol, dimethyl acetainide (DMA), dimethyl formamide (DMF), triacetin, diacetin, corn oil, acetyl triethyl citrate (ATC), ethyl lactate, glycerol formal, ethoxy diglycol (Transcutol, Gattefosse), tryethylene glycol dimethyl ether (Triglylne), dimethyl isosorbide (DMI), y-butyrolactone, N-Methyl-2-pyrrolidinone (NMP), polyethylene glycol of various molecular weights, including but not limited to PEG 300 and PEG
400, and polyglycolated capryl glyceride (Labrasol, Gattefosse), combinations of any one or more of the foregoing, or analogs or derivatives of any one or more of the foregoing.

[0183] In some variations, the solvent is a polyethylene glycol. Polyethylene glycol is known by various names and is available in various preparations, including but not limited to macrogels, macrogel 400, macrogel 1500, macroge14000, macroge16000, macrogel 20000, macrogola, breox PEG; carbowax; carbowax sentry; Hodag PEG;
Lipo;
Lipoxol; Lutrol E; PEG; Pluriol E; polyoxyethylene glycol, and cx Hydro-w-hydroxy-poly(oxy-1,2-ethanediyl).
[01841 In some variations the polyethylene glycol is a liquid PEG, and is one or more of PEG 300 or PEG 400.
[0185] Other solvents include an amount of a C6-C24 fatty acid sufficient to solubilize a therapeutic agent.
[0186] Phospholipid solvents may also be used, such as lecithin, phosphatidylcholine, or a mixture of various diglycerides of stearic, palmitic, and oleic acids, linked to the choline ester of phosphoric acid; hydrogenated soy phosphatidylcholine (HSPC), distearoylphosphatidylglycerol (DSPG), L-a-dimyristoylphosphatidylcholine (DMPC), L-tx-dimyristoylphosphatidylglycerol (DMPG).
[0187] Further examples of solvents include, for example, components such as alcohols, propylene glycol, polyethylene glycol of various molecular weights, propylene glycol esters, propylene glycol esterified with fatty acids such as oleic, stearic, palmic, capric, linoleic, etc; medium chain mono-, di-, or triglycerides, long chain fatty acids, naturally occurring oils, and a mixture thereof. The oily components for the solvent system include commercially available oils as well as naturally occurring oils. The oils may fitrther be vegetable oils or mineral oils. The oils can be characterized as non-surface active oils, which typically have no liydrophile lipophile balance value.
Commercially available substances comprising medium chain triglycerides include, but are not limited to, Captex 100, Captex 300, Captex 355, Miglyol 810, Miglyo1812, Miglyo1818, Miglyol 829, and Dynacerin 660. Propylene glycol ester compositions that are commercially available encompass Captex 200 and Miglyol 840, and the like. The commercial product, Capmul MCM, comprises one of many possible medium chain mixtures comprising monoglycerides and diglycerides.

[0188] Other solvents include naturally occurring oils such as peppermint oil, and seed oils. Exemplary natural oils include oleic acid, castor oil, safflower seed oil, soybean oil, olive oil, sunflower seed oil, sesame oil, and peanut oil. Soy fatty acids may also be used.
Examples of fully saturated non-aqueous solvents include, but are not limited to, esters of medium to long chain fatty acids (such as fatty acid triglycerides with a chain length of about C6 to about C24). Hydrogenated soybean oil and other vegetable oils may also be used. Mixtures of fatty acids may be split from the natural oil (for exainple coconut oil, palm kernel oil, babassu oil, or the like) and refined. In some embodiments, medium chain (about C8 to about C12) triglycerides, such as caprilyic/capric triglycerides derived from coconut oil or palm seed oil, may be used. Medium chain mono- and diglycerides may also be used. Other fully saturated non-aqueous solvents include, but are not limited to, saturated coconut oil (which typically includes a mixture of lauric, myristic, palmitic, capric and caproic acids), including those sold under the MiglyolTM trademark from Huls and bearing trade designations 810, 812, 829 and 840). Also noted are the NeoBeeTM
products sold by Drew Chemicals. Non-aqueous solvents include isopropyl myristate.
Examples of synthetic oils include triglycerides and propylene glycol diesters of saturated or unsaturated fatty acids having 6 to 24 carbon atoms such as, for example hexanoic acid, octanoic (caprylic), nonanoic (pelargonic), decanoic (capric), undecanoic, lauric, tridecanoic, tetradecanoic (myristic), pentadecanoic, hexadecanoic (palmitic), heptadecanoic, octadecanoic (stearic), nonadecanoic, heptadecanoic, eicosanoic, heneicosanoic, docosanoic and lignoceric acids, and the like. Examples of unsaturated carboxylic acids include oleic, linoleic and linolenic acids, and the like.
The non-aqueous solvent can comprise the mono-, di- and triglyceryl esters of fatty acids or mixed glycerides and/or propylene glycol mono- or diesters wherein at least one molecule of glycerol has been esterified with fatty acids of varying carbon atom lengtll.
A non-limiting example of a "non-oil" usefitl as a solvent is polyethylene glycol.
[0189] Exemplary vegetable oils include cottonseed oil, corn oil, sesame oil, soybean oil, olive oil, fractionated coconut oil, peanut oil, sunflower oil, safflower oil, almond oil, avocado oil, palm oil, palm kernel oil, babassu oil, beechnut oil, linseed oil, rape oil and the like. Mono-, di-, and triglycerides of vegetable oils, including but not limited to corn, may also be used.
[01901 Polyvinyl pyrrolidone (PVP), cross-linked or not, may also be used as a solvent.
Further solvents include but are not limited to C6-C24 fatty acids, oleic acid, Imwitor 742, Capmul, F68, F68 (Lutrol), PLURONICS including but not limited to PLURONICS
F108, F127, and F68, Poloxamers, Jeffamines), Tetronics, F127; cyclodextrins such as a-cyclodextrin, 0-cyclodextrin, hydroxypropyl- 0-cyclodextrin, sulfobutylether-cyclodextrin (Captisol); CMC, polysorbitan 20, Cavitron, polyethylene glycol of various molecular weights including but not limited to PEG 300 and PEG 400.

[0191] Beeswax and d-a-tocopherol (Vitainin E) may also be used as solvents.
[0192] Solvents for use in the liquid formulations can be determined by a variety of methods known in the art, including but not limited to (1) theoretically estimating their soh.ibility parameter values and choosing the ones that match with the therapeutic agent, using standard equations in the field; and (2) experimentally determining the saturation solubility of therapeutic agent in the solvents, and choosing the ones that exhibit the desired solubility.

Solubilization of rapanzycin [0193] Where the therapeutic agent is rapamycin, solvents that may be used for making solutions or suspensions of rapamycin include but are not limited to any solvent described herein, including but not limited to any one or more of DMSO, glycerin, ethanol, methanol, isopropyl alcohol; castor oil, propylene glycol, polyvinylpropylene, glycerin, polysorbate 80, benzyl alcohol, dimethyl acetamide (DMA), dimethyl formamide (DMF), --__ glycerol formal, ethoxy diglycol (Transcutol, Gattefosse), tryethylene glycol dimethyl ether (Triglyme), dimethyl isosorbide (DMI), y-butyrolactone, N-Methyl-2-pyrrolidinone (NMP), polyethylene glycol of various molecular weights, including but not limited to PEG 300 and PEG 400, and polyglycolated capryl glyceride (Labrasol, Gattefosse).
[0194] Fur-ther solvents include but are not limited to C6-C24 fatty acids, oleic acid, Imwitor 742, Capmul, F68, F68 (Lutrol), PLURONICS including but not limited to PLURONICS F108, F127, and F68, Poloxamers, Jeffamines), Tetronics, F127, beta-cyclodextrin, CMC, polysorbitan 20, Cavitron, softigen 767, captisol, and sesame oil.
[0195] Other methods that may be used to dissolve rapamycin are described in Solubilization of Rapamycin, P. Simamora et al. Int'/ J. Pliarma 213 (2001) 25-29, the contents of which is incorporated herein in its entirety.

[0196] As a nonlimiting example, rapamycin can be dissolved in 5% DMSO or methanol in a balanced salt solution. The rapamycin solution can be unsaturated, a saturated or a supersaturated solution of rapamycin. The rapamycin solution can be in contact with solid rapamycin. In one nonlimiting example, rapamycin can be dissolved in a concentration of up to about 400 mg/ml. Rapamycin can also, for example, be dissolved in propylene glycol esterified with fatty acids such as oleic, stearic, palmic, capric, linoleic, etc.

[0197] Many other solvents are possible. Those of ordinary skill in the art will find it routine to identify solvents for rapamycin given the teachings herein.

Solubilizing Agents [0198] Generally, any solubilizing agent or combination of solubilizing agents may be used in the liquid formulations described herein.

[0199] In some variations, the solubilizing agent is a surfactant or combination of surfactants. Many surfactants are possible. Combinations of surfactants, including combinations of various types of surfactants, may also be used. For instance, surfactants which are nonionic, anionic (i.e. soaps, sulfonates), cationic (i.e. CTAB), zwitterionic, polymeric or amphoteric may be used.

[0200] Surfactants that can be used may be determined by mixing a therapeutic agent of interest with a putative solvent and a putative surfactant, and observing the characteristics of the formulation after exposure to a medium.

[0201] _ Examples of surfactants include but are not limited to fatty acid esters or amides or ether analogues, or hydrophilic derivatives thereof; monoesters or diesters, or hydrophilic derivatives thereof; or mixtures thereof; monoglycerides or diglycerides, or liydrophilic derivatives thereof; or mixtures thereof; niixtures having enriched mono-or/and diglycerides, or hydrophilic derivatives thereof; surfactants with a partially derivatized with a hydrophilic moiety; monoesters or diesters or multiple-esters of other alcohols, polyols, saccharides or oligosaccharides or polysaccharides, oxyallcylene oligomers or polymers or block polymers, or hydrophilic derivatives thereof, or the amide analogues thereof; fatty acid derivatives of amines, polyamines, polyiinines, aminoalcohols, aminosugars, hydroxyallcylamines, hydroxypolyimines, peptides, polypeptides, or the ether analogues thereof.

10202] Hydrophilic Lipophilic Balance ("HLB") is an expression of the relative simultaneous attraction of a surfactant for water and oil (or for the two phases of the emulsion system being considered).

[0203] Surfactants are characterized according to the balance between the hydrophilic and lipophilic portions of their molecules. The hydrophilic-lipophilic balance (HLB) number indicates the polarity of the molecule in an arbitrazy railge of 1- 40, with the most commonly used emulsifiers having a value between 1- 20. The HLB increases with increasing hydropliilicity.

[0204] Surfactants that may be used include but are not limited to those with an HLB
greater than 10, 11, 12, 13 or 14. Examples of surfactants include polyoxyethylene products of hydrogenated vegetable oils, polyethoxylated castor oils or polyethoxylated hydrogenated castor oil, polyoxyethylene-sorbitan-fatty acid esters, polyoxyethylene castor oil derivatives and the like, for example, Nil&ol HCO-50, Nikkol HCO-35, Niklcol HCO-40, Nikkol HCO-60 (from Nikko Chemicals Co. Ltd.); Cremophor (from BASF) such as Cretnophor RH40, Cremophor RH60, Cremophor EL, TWEENs (from ICI
Chemicals) e.g., TWEEN 20, TWEEN 21, TWEEN 40, TWEEN 60, TWEEN 80, TWEEN 81, Cremophor RH 410, Cremophor RH 455 and the like.

(0205] The surfactant component may be selected from compounds having at least one ether formed from at least about 1 to 100 ethylene oxide units arid at least one fatty alcohol chain having from at least about 12 to 22 carbon atoms; compounds having at least one ester formed from at least about 1 to 100 ethylene oxide units and at least one fatty acid chain having from at least about 12 to 22 carbon atoms; compounds having at least one ether, ester or amide formed from at least about 1 to 100 ethylene oxide units and at least one vitamin or vitamin derivative; and combinations thereof consisting of no more than two surfactants.

[0206] Other examples of surfactants include Lumulse GRH-40, TGPS, Polysorbate-(TWEEN-80), Polysorbate-20 (TWEEN-20), polyoxyethylene (20) sorbitan mono-oleate), glyceiyl glycol esters, polyetliylene glycol esters, polyglycolyzed glycerides, and the like, or mixtures thereof; polyetllylene sorbitan fatty acid esters, polyoxyethylene glycerol esters, such as Tagat TO, Tagat L, Tagat I, tagat 12 and Tagat 0(coininercially available from Goldschmidt Chemical Co., Essen, Germany); ethylene glycol esters, such as glycol stearate and distearate; propylene glycol esters, such as propylene glycol myristate;
glyceryl esters of fatty acids, such as glyceryl stearates and monostearates;
sorbitan esters, such as spans and TWEENs; polyglyceryl esters, such as polyglyceryl 4-oleate;
fatty alcohol ethoxylates, such as Brij type emulsifiers; ethoxylated propoxylated block copolymers, such as poloxamers; polyethylene glycol esters of fatty acids, such as PEG
3001inoleic glycerides or Labrafil 2125 CS, PEG 300 oleic glycerides or Labrafil M 1944 CS, PEG 400 caprylic/capric glycerides or Labrasol, and PEG 300 caprylic/capric glycerides or Softigen 767; cremophors, such as Cremophor E, polyoxyl 35 castor oil or Cremophor EL, Cremophor EL-P, Cremophor RH 40P, polyoxyl 40 hydrogenated castor oil, Cremophor RH40; polyoxyl 60 hydrogenated castor oil or Cremophor RH 60, glycerol monocaprylate/caprate, such as Campmul CM 10; polyoxyethylated fatty acids (PEG-stearates, PED-laurates, Brij ), polyoxylated glycerides of fatty acid, polyoxylated glycerol fatty acid esters i.e. Solutol HS-15; PEG-ethers (Miij(D), sorbitan derivatives (TWEENs), sorbitan monooleate or Span 20, aromatic compounds (Tritons ), PEG-glycerides (PECEOLTM), PEG-PPG (polypropylene glycol) copolymers (PLURONICS
including but not limited to PLURONICS F108, F 127, and F68, Poloxamers, Jeffamines), Tetronics, Polyglycerines, PEG-tocopherols, PEG-LICOL 6-oleate; propylene glycol derivatives, sugar and polysaccharide alkyl and acyl derivatives (octylsucrose, sucrose stearate, laurolydextran etc.) and/or a mixture thereof; surfactants based on an oleate or laureate ester of a polyalcohol copolymerized with ethylene oxide; Labrasol Gelucire 44/14; polyoxytheylene stearates; saturated polyglycolyzed glycerides; or poloxamers; all of which are commercially available. Polyoxyethylene sorbitan fatty acid esters can include polysorbates, for example, polysorbate 20, polysorbate 40, polysorbate 60, and polysorbate 80. Polyoxyethylene stearates can include polyoxyl 6 stearate, polyoxyl 8 stearate, polyoxyl 12 stearate and polyoxyl 20 stearate. Saturated polyglycolyzed glycerides are, for example, GELUCIRE 44/14 or GELUCIRETM 50/13 (Gattefosse, Westwood, N.J., U.S.A.). Poloxainers used herein include poloxamer 124 and poloxamer 188.

[0207] Surfactants include d-cx tocopheryl polyethylene glycol 1000 succinate (TPGS), polyoxyl 8 stearate (PEG 400 monostearate), polyoxy140 stearate (PEG 1750 monostearate) and peppennint oil.

[0208] In some variations, surfactants having an HLB lower than 10 are used.
Such surfactants may optionally be used in combination with other surfactants as co-surfactants.
Examples of some surfactants, mixtures, and other equivalent compositions having an HLB less than or equal to 10 are propylene glycols, glyceryl fatty acids, glyceryl fatty acid esters, polyethylene glycol esters, glyceryl glycol esters, polyglycolyzed glycerides and polyoxyethyl steryl ethers. Propylene glycol esters or partial esters form the composition of commercial products, such as Lauroglycol FCC, which contains propylene glycol laureate. The commercially available excipient Maisine 35-1 comprises long chain fatty acids, for example glyceryl linoleate. Products, such as Acconon E, which comprise polyoxyethylene stearyl ethers, may also be used. Labrafil M 1944 CS is one example of a surfactant wherein the composition contains a mixture of glyceryl glycol esters and polyethylene glycol esters.

Solubiliziug ageitts for rapamyciiz [0209] Many solubilizing agents may be used for rapamycin, including but not limited to those in the solubilizing agents section above.

[0210] In some variations the solubilizing agent is a surfactant. Nonlimiting examples of surfactants that may be used for rapamycin include but are not limited to surfactants with an HLB greater than 10, 11, 12, 13 or 14. One nonlimiting example is Cremophor EL. In some variations, the surfactant may be a polymeric surfactant including but not limited to PLURONICS F108, F127, and F68, and Tetronics. As noted herein, some solvents may also serve as surfactants. Those of ordinary skill in the art will find it routine to identify which solubilizing agents and surfactants may be used for rapamycin given the teachings herein.

Viscosity Modifyiug Ageuts [0211] The liquid fonnulations described herein may be administered with or further comprise a viscosity modifying agent.

[0212] One exemplary viscosity modifying agent that may be used is hyaluronic acid.
Hyaluronic acid is a glycosaminoglycan. It is made of a repetitive sequence of glucuronic acid and glucosamine. Hyaluronic acid is present in many tissues and organs of the body, and contributes to the viscosity and consistency of such tissues and organs.
Hyaluronic acid is present in the eye, inch.iding the vitreous of the eye, and along with collagen contributes to the viscosity thereof. The liquid formulations described herein may further comprise or be administered with hyaluronic acid.

[0213] Other nonlimiting examples of viscosity modifying agents include polyalkylene oxides, glycerol, carboxymethyl cellulose, sodium alginate, chitosan, dextran, dextran sulfate and collagen. These viscosity modifying agents can be chemically modified.
[0214] Otller viscosity modifying agents that may be used include but are not limited to carrageenan, cellulose gel, colloidal silicon dioxide, gelatin, propylene carbonate, carbonic acid, alginic acid, agar, carboxyvinyl polymers or carbomers and polyacrylamides, acacia, ester gum, guar gum, gum arabic, ghatti, gum karaya, tragacanth, terra, pectin, tamarind seed, larch arabinogalactan, alginates, locust bean, xanthan gum, starch, veegum, tragacanth, polyvinyl alcohol, gellan gum, hydrocolloid blends, and povidone. Other viscosity modifying agents known in the art can also be used, including but not limited to sodium carboxymethyl cellulose, algin, carageenans, galactomannans, hydropropyl methyl cellulose, hydroxypropyl cellulose, polyethylene glycol, polyvinylpyrrolidone, sodium carboxymethyl chitin, sodium carboxymethyl dextran, sodium carboxymethyl starch, xanthan gum, and zein.

Other conzpoiients of liquid formulations [0215] The formulations described herein may further comprise various other components such as stabilizers, for example. Stabilizers that may be used in the formulations described herein include but are not limited to agents that will (1) improve the compatibility of excipients with the encapsulating materials such as gelatin, (2) improve the stability (e.g. prevent crystal growth of a therapeutic agent such as rapamycin) of a therapeutic agent such as rapamycin and/or rapamycin derivatives, and/or (3) improve formulation stability. Note that there is overlap between components that are stabilizers and those that are solvents, solubilizing agents or surfactants, and the same component can carry out more than one role.

[0216] Stabilizers may be selected fiom fatty acids, fatty alcohols, alcohols, long chain fatty acid esters, long chain ethers, hydrophilic derivatives of fatty acids, polyvinylpyiTolidones, polyvinylethers, polyvinyl alcohols, hydrocarbons, hydrophobic polymers, moisture-absorbing polymers, and combinations thereof. Amide analogues of the above stabilizers can also be used. The chosen stabilizer may change the hydrophobicity of the formulation (e.g. oleic acid, waxes), or improve the mixing of various components in the fonnulation (e.g. ethanol), control the moisture level in the formula (e.g. PVP), control the mobility of the phase (substances with melting points higher than room temperature such as long chain fatty acids, alcohols, esters, ethers, amides etc. or mixtures thereof; waxes), and/or improve the compatibility of the formula with encapsulating materials (e.g. oleic acid or wax). Some of these stabilizers may be used as solvents/co-solvents (e.g. ethanol). Stabilizers may be present in sufficient amount to inhibit the therapeutic agent's (such as rapamycin's) crystallization.
[0217] Examples of stabilizers include, but are not limited to, saturated, monoenoic, polyenoic, branched, ring-containing, acetylenic, dicarboxylic and fiinctional-group-containing fatty acids such as oleic acid, caprylic acid, capric acid, caproic acid, lauric acid, myristic acid, palmitic acid, stearic acid, behenic acid, linoleic acid, linolenic acid, eicosapentaenoic acid (EPA), DHA; fatty alcohols such as stearyl alcohol, cetyl alcohol, ceteryl alcohol; other alcohols such as ethanol, isopropyl alcohol, butanol;
long chain fatty acid esters, ethers or amides such as glyceryl stearate, cetyl stearate, oleyl ethers, stearyl ethers, cetyl ethers, oleyl amides, stearyl amides; hydrophilic derivatives of fatty acids such as polyglyceryl fatty acids, polyethylene glycol fatty acid esters;
polyvinylpyrrolidones, polyvinylalcohols (PVAs), waxes, docosahexaenoic acid and de-hydroabietic acid etc.
[0218] The forrnulations described may further contain a gelling agent that alters the texture of the final formulation through formation of a gel.
[0219] The therapeutic agents for use as described herein, such as rapamycin, may be subjected to conventional pharmaceutical operations, such as sterilization and compositions containing the therapeutic agent may also contain conventional adjuvants, such as preservatives, stabilizers, wetting agents, emulsifiers, buffers etc.
The therapeutic agents may also be formulated with pharmaceutically acceptable excipients for clinical use to produce a pharmaceutical composition. Formulations for ocular administration may be presented as a solution, suspension, particles of solid material, a discrete mass of solid material, incorporated within a polymer matrix, liquid formulations or in any other form for ocular administration. The therapeutic agents may be used to prepare a medicament to treat, prevent, iidiibit, delay onset, or cause regression of any of the conditions described herein. In some variations, the therapeutic agents may be used to prepare a medicament to treat any of the conditions described herein.

[0220] A coniposition containing a therapeutic agent such as rapainycin may contain one or more adjuvants appropriate for the indicated route of administration.
Adjuvants with which the therapeutic agent may be admixed with include but are not limited to lactose, sucrose, starch powder, cellulose esters of alkanoic acids, stearic acid, talc, magnesium stearate, magnesium oxide, sodium and calcium salts of phosphoric and sulphuric acids, acacia, gelatin, sodium alginate, polyvinylpyrrolidine, and/or polyvinyl alcohol. When a solubilized formulation is required the therapeutic agent may be in a solvent including but not limited to polyethylene glycol of various iuolecular weights, propylene glycol, carboxymethyl cellulose colloidal solutions, methanol, ethanol, DMSO, corn oil, peanut oil, cottonseed oil, sesame oil, tragacanth gum, and/or various buffers.
Other adjuvants and modes of administration are well known in the phanuaceutical art and may be used in the practice of the methods, compositions and liquid formulations described herein. The carrier or diluent may include time delay material, such as glyceryl monostearate or glyceryl distearate alone or with a wax, or other materials well known in the art. The formulations for use as described herein may also include gel formulations, erodible and non-erodible polymers, microspheres, and liposomes.

[0221] Other adjuvants and excipients that may be used include but are not limited to C$-Clo fatty acid esters such as softigen 767, polysorbate 80, PLURONICS, Tetronics, Miglyol, and Transcutol.

[0222] Additives and diluents normally utilized in the pharmaceutical arts can optionally be added to the pharmaceutical composition and the liquid formulation. These include thickening, granulating, dispersing, flavoring, sweetening, coloring, and stabilizing agents, including pH stabilizers, other excipients, anti-oxidants (e.g., tocopherol, BHA, BHT, TBHQ, tocopherol acetate, ascorbyl palmitate, ascorbic acid propyl gallate, and the like), preservatives (e.g., parabens), and the like.
Exemplary preservatives include, but are not limited to, benzylalcohol, ethylalcohol, benzalkonium chloride, phenol, chlorobutanol, and the like. Some useful antioxidants provide oxygen or peroxide inhibiting agents for the formulation and include, but are not limited to, butylated hydroxytoluene, butylhydroxyanisole, propyl gallate, ascorbic acid palmitate, cY
tocopherol, and the like. Thickening agents, such as lecithin, hydroxypropylcellulose, aluminum stearate, and the like, may improve the texture of the formulation.

[0223] In some variations, the therapeutic agent is rapamycin, and the rapamycin is formulated as rapamune in solid or liquid form. In some variations, the rapamune is formulated as an oral dosage.
[0224] Tn addition, a viscous polymer may be added to the suspension, assisting the localization and ease of placement and handling. In some uses of the liquid formulation, a pocket in the sclera may be surgically formed to receive aii injection of the liquid formulations. The hydrogel structure of the sclera can act as a rate-controlling membrane.
Particles of therapeutic agent substance for foiming a suspension can be produced by known methods including but not limited to via ball milling, for example by using ceramic beads. For example, a Cole Parmer ball mill such as Labmill 8000 may be used with 0.8 mm YTZ ceramic beads available from Tosoh or Norstone Inc.

[0225] The formulations may conveniently be presented in unit dosage form and may be prepared by conventional pharmaceutical techniques. Such techniques include the step of bringing into association the therapeutic agent and the pharmaceutical carrier(s) or excipient(s). The formulations may be prepared by uniformly and intimately bringing into associate the active ingredient with liquid carriers or finely divided solid carriers or both, and then, if necessary, shaping the product.
[0226] In some variations, the formulations described herein are provided in one or more unit dose forms, wherein the unit dose form contains an amount of a liquid formulation described herein that is effective to treat or prevent the disease or condition for which it is being administered. In some variations, the formulations described herein are provided in one or more unit dose forms, wherein the unit dose form contains an amount of a liquid rapamycin formulation described herein that is effective to treat or prevent the disease or condition for which it is being administered.

[0227] In some embodiments, the unit dose form is prepared in the concentration at which it will be administered. In some variations, the unit dose form is diluted prior to administration to a subject. In some variations, a liquid formulation described herein is diluted in an aqueous medium prior to administration to a subject. In some variations the aqueous medium is an isotonic medium. In some variations, a liquid formulation described herein is diluted in an non-aqueous medium prior to administration to a subject.

[0228] In a ftirther aspect, provided herein are kits comprising one or more unit dose forms as described herein. In some embodiments, the kit comprises one or more of packaging and instnictions for use to treat one or more diseases or conditions. In some embodiments, the lcit comprises a diluent which is not in physical contact with the formulation or phannaceutical formulation. In some embodiments, the kit comprises any of one or more unit dose forms described herein in one or more sealed vessels.
In some embodiments, the kit comprises any of one or more sterile unit dose forms.
[0229] In some variations, the unit dose form is in a container, including but not limited to a sterile sealed container. In some variations the container is a vial, ampule, or low volume applicator, inch.iding but not limited to a syringe. In some variations, a low-volume applicator is pre-filled with rapamycin for treatment of an ophthalmic disease or condition, including but not limited to a limus compound for treatment of age-related macular degeneration. Described herein is a pre-filled low-volume applicator pre-filled with a formulation comprising a therapeutic agent, including but not limited to rapamycin.
In some variations a low-volume applicator is pre-filled with a solution comprising a therapeutic agent, including but not limited to rapamycin and a polyethylene glycol, and optionally further comprises one or more additional components including but not limited to ethanol. In some variations a pre-filled low-volume applicator is pre-filled with a solution comprising about 2% rapamycin, about 94% PEG-400, about 4% ethanol.
[0230] Described herein are kits comprising one or more containers. In some variations a kit comprises one or more low-volume applicators is pre-filled with a formulation described herein comprising a therapeutic agent, including but not limited to formulations comprising rapamycin, formulations comprising rapamycin and a polyethylene glycol, and optionally further comprises one or more additional components including but not limited to ethanol, and formulations in liquid form comprising about 2% rapamycin, about 94%
PEG-400, about 4% ethanol. In some variations the kit coinprises one or more containers, including but not limited to pre-filled low-voluine applicators, with instructions for its use. In a further variation a kit comprises one or more low-voltime applicators pre-filled with rapamycin, with instnictions for its use in treating a disease or condition of the eye.
In some variations, the containers described herein are in a secondary packaging.

Routes ofAdyrziizistration [0231] The compositions, metliods, and liquid formulations described herein deliver one or more therapeutic agents to a subject, including but not limited to a human subject.
10232] In some variations, the compositions, methods, and liquid foirnulations described herein deliver one or more therapeutic agents to an aqueous medium of a human subj ect.
[0233] In some variations, the compositions, methods, and liquid formulations described herein deliver one or more therapeutic agents to an aqueous medium in or proximal to an area where a disease or conditioil is to be treated, prevented, inhibited, onset delayed, or regression caused.
[0234] In some variations, the compositions, methods, and liquid formulations described herein deliver one or more therapeutic agents to an eye of a subject, including the macula and the retina choroid tissues, in an amount and for a duration effective to treat, prevent, inhibit, delay the onset of, or cause the regression of the diseases and conditions described in the Diseases and Conditions section.
[0235] "Retina choroid" and "retina choroid tissues," as used herein, are synonymous and refer to the combined retina and choroid tissues of the eye.
[0236] As a non-limiting example, the compositions, liquid formulations, and methods described in herein may be administered to the vitreous, aqueous humor, sclera, conjunctiva, between the sclera and conjunctiva, the retina choroid tissues, macula, or other area in or proximate to the eye of a subject, either by direct administration to these tissues or by periocular routes, in amounts and for a duration effective to treat, prevent, inhibit, delay the onset of, or cause the regression of CNV and wet AMD. The effective amounts and durations may be different for each of treating, preventing, inhibiting, delaying the onset of, or causing the regression of CNV and wet AMD, and for each of the different sites of delivery.
[0237] Intravitreal administration is more invasive than some other types of ocular procedures. Because of the potential risks of adverse effects, intravitreal administration may not be optimal for treatment of relatively healthy eyes. By contrast, periocular administration, such as subconjunctival administration, is much less invasive than intravitreal administration. When a therapeutic agent is delivered by a periocular route, it may be possible to treat patients with healthier eyes than could be treated using intravitreal administration. In some variations, subconjunctival injection is used to prevent or delay onset of a disease or condition of the eye, where the eye of the subject has visual acuity of 20/40 or better.

[0238] "Subconjunctival" placement or injection, as used herein, refers to placement or injection between the sclera and conjunctiva. Subconjunctival is sometimes refeiTed to herein as "sub-conj" adininistration.

[0239] Routes of adininistration that may be used to administer a liquid formulation include but are not limited to placement of the liquid formulation, for example by injection, into an aqueous medium in the subject, including but not limited to placement, including but not limited to by injection, into the eye of a subject, including but not limited to a human subject. The liquid formulation may be administered systemically, including but not limited to the following delivery routes: rectal, vaginal, infusion, intrainuscular, intraperitoneal, intraarterial, intrathecal, intrabronchial, intracisternal, cutaneous, subcutaneous, intradermal, transderinal, intravenous, intracervical, intraabdominal, intracranial, intraocular, intrapulmonary, intrathoracic, intratracheal, nasal, buccal, sublingual, oral, parenteral, or nebulised or aerosolized using aerosol propellants.

[0240] Compositions and liquid formulations comprising therapeutic agent can be administered directly to the eye using a variety of procedures, including but not limited to procedures in which (1) the therapeutic agent is administered by injection using a syringe and hypodermic needle, (2) a specially designed device is used to inject the therapeutic agent, (3) prior to injection of the therapeutic agent, a pocket is surgically formed within the sclera to serve as a receptacle for the therapeutic agent or therapeutic agent composition. For example, in one administration procedure a surgeon forms a pocket within the sclera of the eye followed by injection of a solution or liquid formulation comprising the therapeutic agent into the pocket.

[0241] Other administration procedures include, but are not limited to procedures in which (1) a formulation of the therapeutic agent is injected through a specially designed curved cannula to place the therapeutic agent directly against a portion of the eye, (2) a compressed form of the therapeutic agent is placed directly against a portion of the eye, (3) the therapeutic agent is inserted into the sclera by a specially designed injector or inserter, (4) the liquid formulation comprising the therapeutic agent is incorporated within a polymer, (5) a surgeon makes a small conjunctival incision through which to pass a suture and any therapeutic agent delivery structure so as to secure the structure adjacent to the sclera, (6) a needle is used for injection directly into the vitreous of an eye, or into any other site described.

[0242] The liquid formtilations described herein may be used directly, for example, by injection, as an elixir, for topical administration including but not limited to via eye drops, or in hard or soft gelatin or starch capsules. The capsules may be banded to prevent leakage.
Delivery by itajectioya [0243] One method that may be used to deliver the compositions and liquid fonnulations described herein is delivery by injection. In this method compositions and liquid formulations may be injected into a subject, including but not limited to a human subject, or into a position in or proximate to an eye of the subject for delivery to a subject or to the eye of a subject. Injection includes but is not limited to intraocular and periocular injection. Nonlimiting examples of positions that are in or proximate to an eye of a subject are as follows.

[0244] Injection of therapeutic agent into the vitreous may provide a high local concentration of therapeutic agent in the vitreous and retina. Further, it has been found that in the vitreous the clearance half-lives of drugs increases with molecular weight.
[0245] Intracameral injection, or injection into the anterior chamber of they eye, may also be used. In one example, up to about 100 l may be injected intracamerally.

[0246] Periocular routes of delivery may deliver therapeutic agent to the retina without some of the risks of intravitreal delivery. Periocular routes include but are not limited to subconjunctival, subtenon, retrobulbar, peribulbar and posterior juxtascleral delivery. A
"periocular" route of administration means placement near or around the eye.
For a description of exemplary periocular routes for retinal drug delivery, see Pey-ioculan routes for t-etinal drug deliveTy, Raghava et al. (2004), Expert Opin. Drug Deliv.
1(1):99-114, which is incorporated herein by reference in its entirety.

[0247] In some variations the liquid formulations described herein are administered intraocularly. Intraocular administration includes placement or injection within the eye, including in the vitreous.

[0248] Subconjunctival injection may be by injection of therapeutic agent underneath the conjunctiva, or between the sclera and conjunctiva. In one example, up to about 500 gl may be injected subconjunctivally. As one nonlimiting example, a needle of up to about 25 to about 30 gauge and about 30 mm long may be used. Local pressure to the subconjunctival site of therapeutic agent administration may elevate delivery of the therapeutic agent to the posterior segment by reducing local choroidal blood flow.

[0249] Subtenon injection may be by injection of therapeutic agent into the tenon's capsule around the upper portion of the eye and into the "belly" of the superior rectus muscle. In one example, up to about 4 ml may be injected subtenon. As one nonlimiting exainple, a blunt-tipped cannula about 2.5 cm long may be used.
[0250] Retrobulbar injection refers to injection into the conical compartment of the four rectus muscles and their intermuscular septa, behind the globe of the eye. In one example, up to about 5 ml may be injected retrobulbarly. As one nonlimiting example, a blunt needle of about 25 - or about 27-gauge may be used.
[02511 Peribulbar injection maybe at a location external to the confines of the four rectus muscles and their intramuscular septa, i.e., outside of the muscle cone. A volume of, for example, up to about 10 ml maybe injected peribulbarly. As one nonlimiting example, a bh.int-tipped cannula about 1.25 inches long and about 25-gauge may be used.
[0252] Posterior juxtascleral delivery refers to placement of a therapeutic agent near and above the macula, in direct contact with the outer surface of,the sclera, and without puncturing the eyeball. In one example, up to about 500 ml may be injected posterior juxtasclerally. As one nonlimiting example, a blunt-tipped curved cannula, specially designed at 56 , is used to place the therapeutic agent in an incision in the sclera.

[0253] In some variations the liquid formulations described herein are injected intraocularly. Intraocular injection includes injection within the eye.
[0254] Sites to which the compositions and liquid formulations may be administered include but are not limited to the vitreous, aqueous humor, sclera, conjunctiva, between the sclera and conjunctiva, the retina choroid tissues, macula, or other area in or proximate to the eye of a subject. Methods that may be used for placement of the compositions and liquid formulations include but are not limited to injection.

[0255] In one method that may be used, the therapeutic agent is dissolved in an solvent or solvent mixture and then injected into or proximate to the vitreous, aqueous humor, sclera, conjunctiva, between the sclera and conjunctiva, the retina choroid tissues, macula, other area in or proximate to the eye of a subject, or other medium of a subject, according to any of the procedures mentioned above. In one such method that may be used, the therapeutic agent is rapamycin in a liquid formulation.

[0256] When the therapeutic agent is rapamycin, the compositions and liquid formulations may be used to deliver or maintain an ainount of rapamycin in tissues of the eye, including without limitation retina, choroid, or the vitreous, which amount is effective to treat AMD. In one nonlimiting exainple, it is believed that a liquid formulation delivering rapamycin in an amount capable of providing a concentration of rapamycin of about 0.1 pg/mi to about 2 g/ml in the vitreous may be used for treatment of wet AMD. In some nonlimiting examples, it is believed that a liquid formulation delivering a concentration of rapainycin of about 0.1 pg/mg to about 1 .g/mg in the retina choroid tissues may be used for treatment of wet AMD. Other effective concentrations are readily ascertainable by those of skill in the art based on the teachings described herein.

Metlzod ofpreparing liquid formulations [0257] One nonlimiting method that may be used for preparing the liquid formulations described herein, including but not limited to liquid formulations comprising rapamycin, is by mixing a solvent and a therapeutic agent together at room temperature or at slightly elevated temperature until a solution or suspension is obtained, with optional use of a sonicator, and then cooling the formulation. Other components including but not limited to those described above may then be mixed with the formulation. Other preparation methods that may be used are described herein including in the examples, and those of skill in the art will be able to select other preparation methods based on the teachings herein.

Extended delivery of tlaet=apeutic ageuts [0258] Described herein are compositions and liquid formulations showing in vivo delivery or clearance profiles with one or more of the following characteristics. The delivery or clearance profiles are for clearance of the therapeutic agent in vivo after injection of the composition or liquid formulations subconjunctivally or into the vitreous of a rabbit eye. In some variations, the delivery or clearance profiles are for clearance of rapamycin in vivo after injection of the composition or liquid formulations subc_onjunctivally_ or into_the vitreous of a rabbit_eye.~'he volume_of tlLe rabbit vitreous_is__ approximately 30-40% of the volume of the human vitreous. The amount of therapeutic agent is measured using techniques as described in Example 2, but without limitation to the fortnulation and therapeutic agent described in Example 2.

[0259] In some variations, the therapeutic agents with the in vivo delivery or clearance profiles described herein include but are not limited to those described in the T17erapeutic Agents section. In some variations the therapeutic agent is rapamycin. In some variations, the liquid formulations described herein are used to deliver therapeutic agents in a concentration equivalent to rapainycin. The liquid formulations described herein may comprise any therapeutic agent including but not limited to those in the Tlaerapeutic Agents section, in a concentration equivalent to rapamycin including but not limited to those concentrations described herein including in the examples.

[0260] "Average percentage in vivo" level means that an average concentration of therapeutic agent is obtained across multiple rabbit eyes for a given timepoint, and the average concentration of therapeutic agent at one timepoint is divided by the average concentration of therapeutic agent at another timepoint. In some variations of the average percentage in vivo levels, the therapeutic agent is rapamycin.

[0261] The average concentration of a therapeutic agent in the tissue of a rabbit eye at a given time after administration of a formulation containing the therapeutic agent may be measured according to the following method. Where volumes below 10 l are to be injected, a Hamilton syringe is used.

[0262] The liquid formulations are stored at a temperature of 2-8 C prior to use.

[0263] The experimental animals are specific pathogen free (SPF) New Zealand White rabbits. A mixed population of about 50% male, about 50% female is used. The rabbits are at least 12 weelcs of age, usually at least 14 weeks of age, at the time of dosing. The rabbits each weigh at least 2.2 kg, usually at least 2.5 kg, at the time of dosing. Prior to the study, the animals are quarantined for at least one week and examined for general health parameters. Any unhealthy animals are not used in the study. At least 6 eyes are measured and averaged for a given timepoint.

[0264] Housing and sanitation are performed according to standard procedures used in the industry. The animals are provided approximately 150 grams of Teklad Certified Hi-Fiber Rabbit Diet daily, and are provided tap water ad libitum. No contaminants are -_knowsito -existsn the _water._and_no_additional-analysis-outside--that-provided by--the-local ------water district is performed. Environmental Conditions are monitored.

[0265] Each animal undergoes a pre-treatment ophthahnic exainination (slit lamp and ophthalmoscopy), performed by a board certified veterinary ophthalmologist.
Ocular findings are scored according to the McDonald and Shadduck scoring system as described in Dermatoxicology, F. N. Marzulli and H.I. Maibach, 1977 "Eye Irritation,"
T.O.
McDonald and J.A. Shadduck (pages 579-582). Observations are recorded using a standardized data collection sheet. Acceptance criteria for placeinent on study are as follows: scores of <_1 for conjunctival congestion and swelling; scores of 0 for all other observation variables.

[0266] Gentamicin ophthalmic drops are placed into both eyes of each animal twice daily on the day prior to dosing, on the day of dosing (Day 1), and on the day after dosing (Day 2). Dosing is performed in two phases, the first including one set of animals and the second including the other animals. Animals are randomized separately into masked treatment groups prior to each phase of dosing according to modified Latin squares.
Animals are fasted at least 8 hours prior to injection. The start time of the fast and time of inj ection are recorded.

[0267] Animals are weighed and anesthetized with an intravenous injection of a ketamine/xylazine cocktail (87 mg/mL ketamine, 13 mg/mL xylazine) at a volume of 0.1-0.2 mL/kg. Both eyes of each animal are prepared for injection as follows:
approximately five minutes prior to injection, eyes are moistened with an ophthalmic Betadine solution.
After five minutes, the Betadine is washed out of the eyes with sterile saline.
Proparacaine hydrochloride 0.5% (1-2 drops) is delivered to each eye. For eyes to be intravitreally injected, 1% Tropicamide (1 drop) is delivered to each eye.

[0268] On Day 1, both eyes of each animal receive an injection of test or control article.
Animals in selected groups are dosed a second time on Day 90 1. Dosing is subconjunctival or intravitreal. Actual treatments, injection locations, and dose volumes are masked and revealed at the end of the study.

[0269] Subconjunctival injections are given using an insulin syringe and 30 gauge x 1/2-inch needle. The bulbar conjunctiva in the dorsotemporal quadrant is elevated using forceps. Test article is injected into the subconjunctival space.

[0270] Intravitreal injections are given using an Insulin syringe and 30 gauge x 1/2-inch __needle.___For-each inj.ection, the needle is-intr-oduced thr-ough-the-ventr-al-nasal quadr-ant--of---------the eye, approximately 2-3 mm posterior to the limbus, with the bevel of the needle directed downward and posteriorly to avoid the lens. Test article is injected in a single bolus in the vitreous near the retina.

[02711 Animals are observed for mortalityhnorbidity twice daily. Aii animal determined to be moribund is euthanized with an intravenous injection of commercial euthanasia solution. Both eyes are removed and stored frozen at -70 C for possible future evaluation. If an animal is found dead prior to onset of rigor mortis, both eyes are removed and stored frozen at -70 C for possible fitture evaluation. Animals found after the onset of rigor mortis are not necropsied.

[0272] Animals are weighed at randomization, on Day 1 prior to dosing, and prior to euthanasia.

[0273] Ophthalmic observations (slit lamp and indirect ophthalmoscopy) are performed on all animals on Days 5 1, 30 :L 1, 60 1, 90 1, and at later dates in some variations.
Observations are performed by a board certified veterinary ophthalmologist.
For animals to be dosed on Day 90 1, opllthalmic observations are performed prior to dosing.
Ocular findings are scored according to the McDonald and Shadduck scoring system as described in Dermatoxicology, F.N. Marzulli and H.I. Maibach, 1977 "Eye Irritation", T.O. McDonald and J.A. Shadduck (pages 579-582), and observations are recorded using a standardized data collection sheet.

[0274] Whole blood samples (1-3 mL per sample) are collected from each animal prior to necropsy in vacutainer tubes containing EDTA. Each tube is filled at least 2/3 full and thoroughly mixed for at least 30 seconds. Tubes are stored frozen until shipped on dry ice.

[0275] Animals are euthanized with an intravenous injection of commercial euthanasia solution. Euthanasia is performed according to standard procedures used in the industry.
[0276] For treatment groups dosed intravitreally or subconjunctivally with placebo, all eyes from each of these groups are placed into Davidsons solution for approximately 24 hours. Following the 24-hour period, the eyes are transferred to 70% ethanol;
these globes are submitted for masked histopathological evaluation by a board certified veterinary pathologist. The time that eyes are placed into Davidsons and the time of removal are recorded.

__---_--[0277]- -For-treatment_groups-dosedintravitreal-l-y-or-subconjunctival-ly-w-ith test article, --some eyes from each of these groups are frozen at -70 C and submitted for pharmacokinetic analysis. The remaining eyes from each of these groups are placed into Davidsons solution for approximately 24 hours. Following the 24-hour period, the eyes are transferred to 70% ethanol; these globes are submitted for masked histopathological evaluation by a board certified veterinary pathologist. The time that eyes are placed into Davidsons and the time of removal are recorded.

[0278] Frozen sainples submitted for phannacokinetic analysis are dissected with disposable instruments. One set of instniments is used per eye, and then discarded. The samples are thawed at room temperature for 1 to 2 minutes to ensure that the frost around the tissue has been removed. The sclera is dissected into 4 quadrants, and the vitreous is removed. If a non-dispersed mass (NDM) is clearly visible within the vitreous, the vitreous is separated into two sections. The section with the NDM is approximately two-thirds of the vitreous. The section without the NDM is the portion of the vitreous that is the most distant from the NDM. The aqueous humor, lens, iris, and cornea are separated.
The retina choroid tissue is removed using a forceps and collected for analysis. The conjunctiva is separated from the sclera.

[0279] The various tissue types are collected into separate individual pre-weighed vials which are then capped and weighed. The vials of tissue are stored at -80 C
until analyzed.

[0280] The sirolimus content of the retina choroid, sclera, vitreous humor, and whole anti-coagulated blood is determined by high-pressure liquid chromatography/tandem mass spectroscopy (HPLC/MS/MS) using 32-O-desmethoxyrapamycin as an internal standard.
Where an NDM was observed in the vitreous, the section of the vitreous containing the NDM and the section of the vitreous not containing the NDM are analyzed separately.
[0281] The average concentration of a therapeutic agent over a period of time means for representative timepoints over the period of time the average concentration at each time point. For example, if the time period is 30 days, the average concentration may be measured at 5 day intervals: for the average concentration at day 5, the average of a number of measurements of concentration at day 5 would be calculated; for the average concentration at day 10, the average of a number of measurements of the concentration at day 10 would be calculated, etc.

[0282] In some variations, the liquid formulations described herein may have in vivo _--_ -____delivery_to the--vitreous_profiles_with-the.following-described-characteristics,-wher-e-the- --- ---------delivery profiles are for delivery of therapeutic agent in vivo after injection of the liquid foiTnulation between the sclera and the conjunctiva of a rabbit eye. One nonlimiting variation of in vivo deliveiy to the vitreous profiles is shown in Fig. 2.

[0283] At day 40 after injection, the average percentage in vivo vitreal level may be between about 70% and about 100%, and more usually between about 80% and about 90%, relative to the level present at day 20 after injection. At day 40 after injection, the average percentage in vivo vitreal level may be greater than about 70%, and more usually greater than about 80%, relative to the level present at day 20 after injection.

[0284] At day 67 after injection, the average percentage in vivo vitreal level may be between about 75% and about 115%, and more usually between about 85% and about 105%, relative to the level present at day 20 after injection. At day 67 after injection, the average percentage in vivo vitreal level may be greater than about 75%, and more usually greater than about 85%, relative to the level present at day 20 after injection.

[0285] At day 90 after injection, the average percentage in vivo vitreal level may be between about 20% and about 50%, and more usually between about 30% and about 40%, relative to the level present at day 20 after injection. At day 90 after injection, the average percentage in vivo vitreal level may be greater than about 20%, and more usually greater than about 30%, relative to the level present at day 20 after injection.

[0286] In some variations, the average percentage in vivo vitreal level has the following characteristics relative to the level present at day 20 after injection: at 40 days after injection it is less than about 100%; at 67 days after injection it is less than about 115%;
and 90 days after injection it is less than about 50%.

[0287] In some variations, the liquid formulation when injected between the sclera and conjunctiva of a rabbit eye delivers therapeutic agent giving an average concentration of therapeutic agent in the vitreous of the rabbit eye of at least about 0.01 ng/mL for at least about 30, at least about 60, or at least about 90 days after administration of the liquid formulation to the rabbit eyes. In some variations, the liquid formulation when injected between the sclera and conjunctiva of a rabbit eye delivers therapeutic agent giving an average concentration of therapeutic agent in the vitreous of the rabbit eye of at least -about 0.1 ng/inL for at least about 30, at least about 60, or at least about 90 days after administration of the liquid formulation to the rabbit eyes. In some variations, the liquid ___ ----- _._...formulation wheninj.ectedbetween..thesclera_and conjunctiva_ofa_rabb-it_ey_e_delivers_ _- __ ___ therapeutic agent giving an average concentration of therapeutic agent in the vitreous of the rabbit eye of at least about 1 ng/mL for at least about 30, at least about 60, or at least about 90 days after administration of the liquid formulation to the rabbit eyes.

[0288] In some variations, the liquid formulations described herein may have in vivo delivery to the retina choroid profiles with the following described characteristics, where the delivery profiles are for delivery of therapeutic agent in vivo after injection of the liquid formulation between the sclera and the conjunctiva of a rabbit eye.

[0289] At day 40 after injection, the average percentage in vivo retina choroid level may be between about 350% and about 410%, and more usually between about 360% and about 400%, relative to the level present at day 20 after injection. At day 40 after inj ection, the average percentage in vivo retina choroid level may be greater than about 350%, and more usually greater than about 360%, relative to the level present at day 20 after injection.

[0290] At day 67 after injection, the average percentage in vivo retina choroid level may be between about 125% and about 165%, and more usually between about 135% and about 155%, relative to the level present at day 20 after injection. At day 67 after injection, the average percentage in vivo retina choroid level may be greater than about 125%, and more usually greater than about 135%, relative to the level present at day 20 after injection.
[0291] At day 90 after injection, the average percentage in vivo retina choroid level may be between about 10% and about 50%, and more usually between about 20% and about 40%, relative to the level present at day 20 after injection. At day 90 after injection, the average percentage in vivo retina choroid level may be greater than about 10%, and more usually greater than about 20%, relative to the level present at day 20 after injection.
[0292] In some variations, the average percentage in vivo retina choroid level has the following characteristics relative to the level present at day 20 after injection: at 40 days after injection it is less than about 410%; at 67 days after injection it is less than about 165%; and 90 days after injection it is less than about 50%.

[0293] In some variations, the liquid formulation when injected between the sclera and conjunctiva of a rabbit eye delivers therapeutic agent giving an average concentration of therapeutic agent in the retina choroid tissues of the rabbit eye of at least about 0.001 ----ng/mg-for-at-least about 30,- at .least about_60,_or-at-least-about 90--days_ after_.administration_ of the liquid formulation to the rabbit eyes. In some variations, the liquid formulation when injected between the sclera and conjunctiva of a rabbit eye delivers therapeutic agent giving an average concentration of therapeutic agent in the retina choroid tissues of the rabbit eye of at least about 0.01 ng/ing for at least about 30, at least about 60, or at least about 90 days after administration of the liquid formulation to the rabbit eyes.

[0294] In some variations, the level of therapeutic agent present in the retina choroid first increases, then peaks and decreases. The pealc may, for instance, occur at about day 40 after injection.

[0295] In some variations, the liquid formulations described herein may have in vivo clearance from the sclera profiles with the following described characteristics, where the clearance profiles are for clearance of therapeutic agent in vivo after injection of the liquid formulation between the sclera and the conjunctiva of a rabbit eye. Where injection is between the sclera and the conjunctiva, the scleral level is thought to include the injected liquid fonnulation.

[0296] At day 40 after injection, the average percentage in vivo scleral level may be between about 150% and about 230%, and more usually between about 170% and about 210%, relative to the level present at day 20 after injection. At day 40 after injection, the average percentage in vivo scleral level maybe greater than about 150%, and more usually greater than about 170%, relative to the level present at day 20 after injection.
[0297] At day 67 after injection, the average percentage in vivo scleral level may be between about 30% and about 70%, and more usually between about 40% and about 60%, relative to the level present at day 20 after injection. At day 67 after injection, the average percentage in vivo scleral level maybe greater than about 30%, and more usually greater than about 40%, relative to the level present at day 20 after injection.

[0298] At day 90 after injection, the average percentage in vivo scleral level maybe between about 110% and about 160%, and more usually between about 125% and about 145%, relative to the level present at day 20 after injection. At day 90 after injection, the average percentage in vivo scleral level may be greater than about 110%, and more usually greater than about 125%, relative to the level present at day 20 after injection.
[0299] In some variations, the average percentage in vivo scieral level has the following characteristics relative to the level present at day 20 after injection: at 40 days after injection it is less than about 230%; at 67 days after injection it is less than about 70%;
and 90 days after injection it is less than about 160%0._ [0300] In some variations, the level of therapeutic agent present in the sclera first increases, then peaks and decreases. The pealc may, for instance, occur at about day 40 after injection.

[0301] In some variations, the liqtiid formulations described herein may have in vivo delivery to the vitreous profiles with the following described characteristics, where the delivery profiles are for delivery of therapetitic agent in vivo after injection of the liquid formulation between the sclera and the conjunctiva of a rabbit eye.

[0302] At day 14 after injection, the average percentage in vivo vitreal level may be between about 1350% and about 1650%, and more usually between about 1450% and abottt 1550%, relative to the level present at day 2 after injection. At day 14 after injection, the average percentage in vivo vitreal level may be greater than about 1350%, and more usually greater thal about 1450%, relative to the level present at day 2 after injection.

[0303] At day 35 after injection, the average percentage in vivo vitreal level may be between about 200% and about 300%, and more usually between about 225% and about 275%, relative to the level present at day 2 after injection. At day 35 after injection, the average percentage in vivo vitreal level may be greater than about 200%, and more usually greater than about 225%, relative to the level present at day 2 after injection.
[0304] At day 62 after injection, the average percentage in vivo vitreal level may be between about 100% and about 160%, and more usually between abotit 115% and about 145%, relative to the level present at day 2 after injection. At day 62 after injection, the average percentage in vivo vitreal level may be greater than about 100%, and more usually greater than about 115%, relative to the level present at day 2 after injection.
[0305] At day 85 after injection, the average percentage in vivo vitreal level may be between about 5% and about 30%, and more usually between about 10% and about 25%, relative to the level present at day 2 after injection. At day 85 after injection, the average percentage in vivo vitreal level may be greater than about 5%, and more usually greater than about 10%, relative to the level present at day 2 after injection.

[0306] In some variations, the average percentage in vivo vitreal level has the following characteristics relative to the level present at day 2 after injection: at 14 days after injection it is less than about 1600%; at 35 days afterlnjection it is less than about_300o;__ - at 62 days after injection it is less than about 160% and 85 days after injection it is less than about 30%.

[0307] In some variations, the liquid formulation when injected between the sclera and conjunctiva of a rabbit eye delivers therapeutic agent giving an average concentration of therapeutic agent in the vitreous of the rabbit eye of at least about 0.01 ng/inL for at least about 30, at least about 60, or at least about 85 days after administration of the liquid formulation to the rabbit eyes. In some variations, the liquid formulation when injected between the sclera and conjunctiva of a rabbit eye delivers therapeutic agent giving an average concentration of therapeutic agent in the vitreous of the rabbit eye of at least about 0.1 ng/mL for at least about 30, at least about 60, or at least about 85 days after administration of the liquid formulation to the rabbit eyes. In some variations, the liquid formulation when injected between the sclera and conjunctiva of a rabbit eye delivers therapeutic agent giving an average concentration of therapeutic agent in the vitreous of the rabbit eye of at least about 1 ng/mL for at least about 30, or at least about 60 days after administration of the liquid formulation to the rabbit eyes.

[0308] In some variations, the level of therapeutic agent present in the vitreous first increases, then peaks and decreases. The peak may, for instance, occur at about day 14 after injection.

[0309] In some variations, the liquid formulations described herein may have in vivo delivery to the retina choroid profiles with the following described characteristics, where the delivery profiles are for delivery of therapeutic agent in vivo after injection of the liquid formulation between the sclera and the conjunctiva of a rabbit eye.

[0310] At day 35 after injection, the average percentage in vivo retina choroid level may be between about 320% and about 400%, and more usually between about 340% and about 380%, relative to the level present at day 14 after injection. At day 35 after injection, the average percentage in vivo retina choroid level may be greater than about 320%, and more usually greater than about 340%, relative to the level present at day 14 after injection.

[0311] At day 62 after injection, the average percentage in vivo retina choroid level may be between about 3% and about 25%, and more usually between about 6% and about 20%, relative to the level present at day 14 after injection. At day 62 after injection, the average percentage in vivo retina choroid level may be greater than about 3%, and more __usuallygr.eater_than_ab.o.ut_6%o,_relative.to_the_level_presentat_da_y__14aft er_inj.ection.________ [0312] At day 85 after injection, the average percentage in vivo retina choroid level may be between about 0.1% and about 6%, and more usually between about 0.5% and about 4%, relative to the level present at day 14 after injection. At day 85 after injection, the average percentage in vivo retina choroid level may be greater than about 0.1 %, and more usually greater than about 0.5%, relative to the level present at day 14 after injection.
[0313] In some variations, the average percentage in vivo retina choroid level has the following characteristics relative to the level present at day 14 after injection: at 35 days after injection it is less than about 400%; at 62 days after injection it is less than about 25%; and 85 days after injection it is less than about 6%.

(0314] In some variations, the liquid formulation when injected between the sclera and conjunctiva of a rabbit eye delivers therapeutic agent giving an average concentration of therapeutic agent in the retina choroid tissues of the rabbit eye of at least about 0.001 ng/mg for at least about 30, at least about 60, or at least about 85 days after administration of the liquid formulation to the rabbit eyes. In some variations, the liquid formulation when injected between the sclera and conjunctiva of a rabbit eye delivers therapeutic agent giving an average concentration of therapeutic agent in the retina choroid tissues of the rabbit eye of at least about 0.01 ng/mg for at least about 30, at least about 60, or at least about 85 days after administration of the liquid fonnulation to the rabbit eyes.

[0315] In some variations, the liquid formulations described herein may have in vivo clearance from the sclera profiles with the following described characteristics, where the clearance profiles are for clearance of therapeutic agent in vivo after injection of the liquid formulation between the sclera and the conjunctiva of a rabbit eye. For injection between the sclera and conjunctiva, the scleral level is thought to include the injected liquid formulation.
[0316] At day 35 after injection, the average percentage in vivo scleral level may be between about 0.1% and about 0.7%, and more usually between about 0.2% and about 0.6%, relative to the level present at day 14 after injection. At day 35 after injection, the average percentage in vivo scleral level may be greater than about 0.1 %, and more usually greater than about 0.2%, relative to the level present at day 14 after injection.

(0317] At day 62 after injection, the average percentage in vivo scleral level may be between about 0.05% and about 0.35%, and more usually between about 0.07% and about 0.3%, relative to the level present at day 14 after injection. At day 62 after injection, the average percentage in vivo scleral level may be greater than about 0.05%, -and-more_ usually greater than about 0.07%, relative to the level present at day 14 after injection.

[0318] At day 85 after injection, the average percentage in vivo scleral level may be between about 0.1% and about 0.9%, and more usually between about 0.3% and about 0.7%, relative to the level present at day 14 after injection. At day 85 after injection, the average percentage in vivo scleral level may be greater than about 0.1 %, and more usually greater than about 0.3%, relative to the level present at day 14 after injection.

[0319] In some variations, the average percentage in vivo scleral level has the following characteristics relative to the level present at day 14 after injection: at 35 days after injection it is less than about 0.7%; at 62 days after injection it is less than about 0.35%;
and 85 days after injection it is less tha.n about 0.9%.

[0320] In some variations, the liquid formulations described herein may have in vivo clearance from the vitreous profiles with the following described characteristics, where the clearance profiles are for clearance of therapeutic agent in vivo after injection of the liquid formulation into the vitreous of a rabbit eye. Where injection is into the vitreous, the measured vitreous level is thought to include the injected formulation.

[0321] At day 35 after injection, the average percentage in vivo vitreal level maybe between about 1% and about 40%, and more usually between about 1% and about 10%, relative to the level present at day 14 after injection. At day 35 after injection, the average percentage in vivo vitreal level may be greater than about 1% relative to the level present at day 14 after injection.

[0322] At day 62 after injection, the average percentage in vivo vitreal level may be between about 1% and about 40%, and more usually between about 5% and about 25%, relative to the level present at day 14 after injection. At day 62 after injection, the average percentage in vivo vitreal level may be greater than about 1% relative to the level present at day 14 after injection, and more usually greater than about 5% relative to the level present at day 14 after injection.

[0323] At day 90 after injection, the average percentage in vivo vitreal level may be between about 1% and about 40%, and more usually between about 10% and about 30%, relative to the level present at day 14 after injection. At day 90 after injection, the average percentage in vivo vitreal level may be greater than about 1% relative to the level present at day 14 after injection, and more usually greater than about 10% relative to the level present at day 14 after injection.

[0324] In some variations, the level of therapeutic agent present in the vitreous first increases, then peaks and decreases. The pealc may, for instance, occur at about day 14 after injection.

[0325] In some variations, the liquid fonnulations described herein may have in vivo delivery to the retina choroid profiles with the following described characteristics, where the delivery profiles are for delivery of therapeutic agent in vivo after injection of the liquid formulation into the vitreous of a rabbit eye.

[0326] At day 35 after injection, the average percentage in vivo retina choroid level may be between about 3400% and about 5 100%, and more usually between about 3750%
and about 4750%, relative to the level present at day 14 after injection. At day 35 after injection, the average percentage in vivo retina choroid level may be greater than about 3400%, and more usually greater than about 3750%, relative to the level present at day 14 after injection.

[0327] At day 62 after injection, the average percentage in vivo retina choroid level may be between about 0.1 % and about 5%, and more usually between about 1% and about 3%, relative to the level present at day 14 after injection. At day 62 after injection, the average percentage in vivo retina choroid level may be greater than about 0.1 %, and more usually greater than about 1%, relative to the level present at day 14 after injection.

[0328] At day 90 after injection, the average percentage in vivo retina choroid level may be between about 10% and about 50%, and more usually between about 20% and about 40%, relative to the level present at day 14 after injection. At day 90 after injection, the average percentage in vivo retina choroid level may be greater than about 10%, and more usually greater than about 20%, relative to the level present at day 14 after injection.
[0329] In some variations, the average percentage in vivo retina choroid level has the following characteristics relative to the level present at day 14 after injection: at 35 days after injection it is less than about 5 100%; at 62 days after injection it is less than about 5%; and 90 days after injection it is less than about 50%.

[0330] In some variations, the liquid formulations described herein may have in vivo delivery to the sclera profiles with the following described characteristics, where the delivery profiles are for delivery of therapeutic agent in vivo after injection of the liquid formulation into the vitreous of a rabbit eye.
------------_-----[0331] At day 35 after injection, the average percentage in vivo scleral level may be between about 1700% and about 2600%, and more usually between about 1900% and about 2400%, relative to the level present at day 14 after injection. At day 35 after injection, the average percentage in vivo scleral level may be greater than about 1700%, and more usually greater than about 1900%, relative to the level present at day 14 after injection.
[0332] At day 62 after injection, the average percentage in vivo scleral level may be between about 120% and about 180%, and more usually between about 140% and about 160%, relative to the level present at day 14 after injection. At day 62 after injection, the average percentage in vivo scleral level may be greater than about 120%, and more usually greater than about 140%, relative to the level present at day 14 after injection.
[0333] At day 90 after injection, the average percentage in vivo scleral level may be between about 95% and about 155%, and more usually between about 115% and about 135%, relative to the level present at day 14 after injection. At day 90 after injection, the average percentage in vivo scleral level may be greater than about 95%, and more usually greater than about 115%, relative to the level present at day 14 after injection.

[0334] In some variations, the average percentage in vivo scleral level has the following characteristics relative to the level present at day 14 after injection: at 35 days after injection it is less than about 2600%; at 62 days after injection it is less than about 180%;
and 90 days after injection it is less than about 155%.

[0335] In some variations, the liquid fonnulation when injected into the vitreous of a rabbit eye delivers therapeutic agent giving an average concentration of therapeutic agent in the sclera of the rabbit eye of at least about 0.001 ng/mg for at least about 30, at least about 60, or at least about 90 days after administration of the liquid fonnulation to the rabbit eyes. In some variations, the liquid formulation when injected into the vitreous of a rabbit eye delivers therapeutic agent giving an average concentration of therapeutic agent in the sclera of the rabbit eye of at least about 0.01 ng/mg for at least about 30, at least about 60, or at least about 90 days after administration of the liquid formulation to the rabbit eyes. In some variations, the liquid formulation when injected into the vitreous of a rabbit eye delivers therapeutic agent giving an average concentration of therapeutic agent in the sclera of the rabbit eye of at least about 0.1 ng/mg for at least about 30, at least about 60, or at least about 90 days after administration of the liquid fonnulation to the [0336] In some variations, the level of therapeutic agent present in the vitreous first increases, then peaks and decreases. The peak may, for instance, occur at about day 35 after injection.

[0337] In some variations, in situ gelling liquid formulations described herein may have in vivo deliveiy to the vitreous profiles with the following described characteristics, where the delivery profiles are for delivery of therapeutic agent in vivo after injection of the liquid formulation between the sclera and the conjunctiva of a rabbit eye.

[0338] At day 32 after injection, the average percentage in vivo vitreal level may be between about 25% and about 85%, and more usually between about 45% and about 65%, relative to the level present at day 7 after injection. At day 40 after injection, the average percentage in vivo vitreal level may be greater than about 25%, and more usually greater than about 45%, relative to the level present at day 7 after injection.

[0339] At day 45 after injection, the average percentage in vivo vitreal level may be between about 2% and about 50%, and more usually between about 8% and about 20%, relative to the level present at day 7 after injection. At day 67 after injection, the average percentage in vivo vitreal level may be greater than about 2%, and more usually greater than about 5%, relative to the level present at day 7 after injection.

[0340] At day 90 after injection, the average percentage in vivo vitreal level may be between about 40% and about 100%, and more usually between about 60% and about 80%, relative to the level present at day 7 after injection. At day 90 after injection, the average percentage in vivo vitreal level may be greater than about 40%, and more usually greater than about 60%, relative to the level present at day 7 after injection.
[0341] In some variations, the average percentage in vivo vitreal level has the following characteristics relative to the level present at day 7 after injection: at 32 days after injection it is less than about 80%; at 45 days after injection it is less than about 30%; and 90 days after injection it is less than about 100%.

[0342] In some variations, the liquid formulation when injected between the sclera and conjunctiva of a rabbit eye delivers therapeutic agent giving an average concentration of therapeutic agent in the vitreous of the rabbit eye of at least about 0.1 pg/mL for at least about 30, at least about 60, or at least about 90 days after administration of the liquid -----formulation-to-the-rabbit-eye. In some-var-iations, the-liquid-formulation when injected-------------between the sclera and conjunctiva of a rabbit eye delivers therapeutic agent giving an average concentration of therapeutic agent in the vitreous of the rabbit eye of at least about 0.01 ng/mL for at least about 30, at least about 60, or at least about 90 days after administration of the liquid formulation to the rabbit eye. In some variations, the liquid foimulation when injected between the sclera and conjunctiva of a rabbit eye delivers therapeutic agent giving an average concentration of therapeutic agent in the vitreous of the rabbit eye of at least about 0.1 ng/mL for at least about 30, at least about 60, or at least about 90 days after administration of the liquid formulation to the rabbit eye. In some variations, the liquid formulation when injected between the sclera and conjunctiva of a rabbit eye delivers therapeutic agent giving an average concentration of therapeutic agent in the vitreous of the rabbit eye of at least about 1 ng/inL for at least about 30, at least about 60, or at least about 90 days after administration of the liquid forinulation to the rabbit eye. In some variations, the liquid formulation when injected between the sclera and conjunctiva of a rabbit eye delivers therapeutic agent giving an average concentration of therapeutic agent in the vitreous of the rabbit eye of at least about 10 ng/mL for at least about 30, at least about 60, or at least about 90 days after administration of the liquid formulation to the rabbit eye.

[0343] In some variations, the liquid formulation when injected between the sclera and conjunctiva of a rabbit eye delivers therapeutic agent giving an average concentration of therapeutic agent in the vitreous of the rabbit eye of at least 0.001 ng/mL
for at least 30, at least 60, at least 90, or at least 120 days after administration of the liquid formulation to the rabbit eyes. In some variations, the liquid formulation when injected between the sclera and conjunctiva of a rabbit eye delivers therapeutic agent giving an average concentration of therapeutic agent in the vitreous of the rabbit eye of at least 0.01 ng/mL
for at least 30, at least 60, at least 90, or at least 120 days after administration of the liquid formulation to the rabbit eyes. In some variations, the liquid formulation when injected between the sclera and conjunctiva of a rabbit eye delivers therapeutic agent giving an average concentration of therapeutic agent in the vitreous of the rabbit eye of at least 0.1 ng/mL for at least 30, at least 60, at least 90, or at least 120 days after administration of the liquid formulation to the rabbit eyes. In some variations, the liquid formulation when injected between the sclera and conjunctiva of a rabbit eye delivers therapeutic agent giving an average concentration of therapeutic agent in the vitreous of the rabbit eye of at least 0.5 ng/mL for at least 30, at least 60, at least 90, or at least 120 days after administration of the liquid formulation to the rabbit eyes.

[0344] In some variations, the liquid formulation when injected between the sclera and conjunctiva of a rabbit eye delivers therapeutic agent giving an average concentration of therapeutic agent in the vitreous of the rabbit eye of between 0.001 ng/mL and 10.0 ng/mL
for at least 30, at least 60, at least 90, or at least 120 days after administration of the liquid formulation to the rabbit eyes. In some variations, the liquid formulation when injected between the sclera and conjunctiva of a rabbit eye delivers therapeutic agent giving an average concentration of therapeutic agent in the vitreous of the rabbit eye of between 0.01 ng/mL and 10 ng/mL for at least 30, at least 60, at least 90, or at least 120 days after administration of the liquid formulation to the rabbit eyes. In some variations, the liquid forinulation when injected between the sclera and conjunctiva of a rabbit eye delivers therapeutic agent giving an average concentration of therapeutic agent in the vitreous of the rabbit eye of between 0.1 ng/inL and 10 ng/mL for at least 30, at least 60, at least 90, or at least 120 days after administration of the liquid formulation to the rabbit eyes.
[03451 In some variations, the liquid formulation when injected between the sclera and conjunctiva of a rabbit eye delivers therapeutic agent giving an average concentration of therapeutic agent in the vitreous of the rabbit eye of between 0.5 ng/mL and 10.0 ng/mL
for at least 30, at least 60, at least 90, or at least 120 days after administration of the liquid formulation to the rabbit eyes.
[0346] In some variations, the liquid formulation when injected between the sclera and conjunctiva of a rabbit eye delivers therapeutic agent giving a ratio of a maximum average concentration of therapeutic agent in the vitreous of a rabbit eye to a minimum average concentration of therapeutic agent in the vitreous of a rabbit eye less than 100 for days 30 to at least 60, at least 90, or at least 120 days after administration of the liquid formulation to the rabbit eyes. In some variations, the liquid formulation when injected between the sclera and conjunctiva of a rabbit eye delivers therapeutic agent giving a ratio of a maximum average concentration of therapeutic agent in the vitreous of a rabbit eye to a minimum average concentration of therapeutic agent in the vitreous of a rabbit eye less than 50 for days 30 to at least 60, at least 90, or at least 120 days after administration of the liquid formulation to the rabbit eyes. In some variations, the liquid formulation when injected between the sclera and conjunctiva of a rabbit eye delivers therapeutic agent giving a ratio of a maximum average concentration of therapeutic agent in the vitreousof a rabbit eye to a minimum average concentration of therapeutic agent in the vitreous of a rabbit eye less than 10 for days 30 to at least 60, at least 90, or at least 120 days after administration of the liquid forinulation to the rabbit eyes. In some variations, the liquid fonnulation when injected between the sclera and conjunctiva of a rabbit eye delivers therapeutic agent giving a ratio of a maximum average concentration of therapeutic agent in the vitreous of a rabbit eye to a minimum average concentration of therapeutic agent in the vitreous of a rabbit eye less than 5 for days 30 to at least 60, at least 90, or at least 120 days after administration of the liquid formulation to the rabbit eyes.

[0347) "Approximately constant," as used herein, means that the average level does not vary by more than one order of magnitude over the extended period of time, i.e., the difference between the maximum and minimum is less than a 10-fold difference for measurements of the average concentration at times in the relevant period of time.

[0348] In some variations, the liquid forinulation when injected between the sclera and conjunctiva of a rabbit eye delivers therapeutic agent giving an average concentration of therapeutic agent in the vitreous of a rabbit eye that is approximately constant at a value greater than 0.00 1 ng/mL for days 30 to at least 60, at least 90, or at least 120 days after administration of the solution to the rabbit eyes. In some variations, the liquid formulation when injected between the sclera and conjunctiva of a rabbit eye delivers therapeutic agent giving an average concentration of therapeutic agent in the vitreous of a rabbit eye that is approximately constant at a value greater than 0.01 ng/mL
for days 30 to at least 60, at least 90, or at least 120 days after administration of the liquid formulation to the rabbit eyes. In some variations, the liquid formulation when injected between the sclera and conjunctiva of a rabbit eye delivers therapeutic agent giving an average concentration of therapeutic agent in the vitreous of a rabbit eye that is approximately constant at a value greater than 0.1 ng/mL for days 30 to at least 60, at least 90, or at least 120 days after administration of the liquid formulation to the rabbit eyes. In some variations, the liquid formulation when injected between the sclera and conjunctiva of a rabbit eye delivers therapeutic agent giving an average concentration of therapeutic agent in the vitreous of a rabbit eye that is approximately constant at a value of 1.0 ng/mL for days 30 to at least 60, at least 90, or at least 120 days after administration of the liquid formulation to the rabbit eyes.

[0349] In some variations, the liquid fonnulation when injected between the sclera and conjunctiva of a rabbit eye delivers therapeutic agent giving an average concentration of_ -__--- therapeutic agent in the retina choroid tissues of the rabbit eye of at least 0.001 ng/mg for at least 30, at least 60, at least 90, or at least 120 days after adniinistration of the liquid fonnulation to the rabbit eyes. In some variations, the liquid formulation when injected between the sclera and conjunctiva of a rabbit eye delivers therapeutic agent giving an average concentration of therapeutic agent in the retina choroid tissues of the rabbit eye of at least 0.005 ng/mg for at least 30, at least 60, at least 90, or at least 120 days after administration of the liquid fonnulation to the rabbit eyes. In some variations, the liquid formulation when injected between the sclera and conjunctiva of a rabbit eye delivers therapeutic agent giving an average concentration of therapeutic agent in the retina choroid tissues of the rabbit eye of at least 0.01 ng/mg for at least 30, at least 60, at least 90, or at least 120 days after administration of the liquid formulation to the rabbit eyes.
[0350] In some variations, the liquid formulation when injected between the sclera and conjunctiva of a rabbit eye delivers therapeutic agent giving an average concentration of therapeutic agent in the retina choroid tissues of the rabbit eye of between 0.001 ng/ing and 1.0 ng/mg for at least 30, at least 60, at least 90, or at least 120 days after administration of the liquid formulation to the rabbit eyes. In some variations, the liquid formulation when injected between the sclera and conjunctiva of a rabbit eye delivers therapeutic agent giving an average concentration of therapeutic agent in the retina choroid tissues of the rabbit eye of between 0.001 ng/mg and 0.50 ng/mg for at least 30, at least 60, at least 90, or at least 120 days after administration of the liquid formulation to the rabbit eyes. In some variations, the liquid formulation when injected between the sclera and conjunctiva of a rabbit eye delivers therapeutic agent giving an average concentration of therapeutic agent in the retina choroid tissues of the rabbit eye of between 0.001 ng/mg and 0.15 ng/mg for at least 30, at least 60, at least 90, or at least 120 days after administration of the liquid formulation to the rabbit eyes. In some variations, the liquid forinulation when injected between the sclera and conjunctiva of a rabbit eye delivers therapeutic agent giving an average concentration of therapeutic agent in the retina choroid tissues of the rabbit eye of between 0.001 ng/mg and 0.1 ng/mg for at least 30, at least 60, at least 90, or at least 120 days after administration of the liquid formulation to the rabbit eyes.

[0351] In some variations, the liquid formulation when injected between the sclera and conjunctiva of a rabbit eye delivers therapeutic agent giving an average concentration of therapeutic agent in the Tetina_choroid_tissues of the rabbit eye ofbetween_0.0-05_ng/mg__.
and 1.0 ng/mg for at least 30, at least 60, at least 90, or at least 120 days after administration of the liquid formulation to the rabbit eyes. In some variations, the liquid fonnulation when injected between the sclera and conjunctiva of a rabbit eye delivers therapeutic agent giving an average concentration of therapeutic agent in the retina choroid tissues of the rabbit eye of between 0.005 ng/hng and 0.50 ng/mg for at least 30, at least 60, at least 90, or at least 120 days after administration of the liquid formulation to the rabbit eyes. In some variations, the liquid formulation when injected between the sclera and conjuilctiva of a rabbit eye delivers therapeutic agent giving an average concentration of therapeutic agent in the retina choroid tissues of the rabbit eye of between 0.005 ng/mg and 0.15 ng/ing for at least 30, at least 60, at least 90, or at least 120 days after administration of the liquid fonnulation to the rabbit eyes. In some variations, the liquid formulation when injected between the sclera and conjunctiva of a rabbit eye delivers tlierapeutic agent giving an average concentration of therapeutic agent in the retina choroid tissues of the rabbit eye of between 0.005 ng/mg and 0.1 ng/mg for at least 30, at least 60, at least 90, or at least 120 days after administration of the liquid formulation to the rabbit eyes.

[0352] In some variations, the liquid fonnulation when injected between the sclera and conjunctiva of a rabbit eye delivers therapeutic agent giving an average concentration of therapeutic agent in the retina choroid tissues of the rabbit eye of between 0.01 ng/mg and 1.0 ng/mg for at least 30, at least 60, at least 90, or at least 120 days after administration of the liquid fonnulation to the rabbit eyes. In some variations, the liquid fonnulation when injected between the sclera and conjunctiva of a rabbit eye delivers therapeutic agent giving an average concentration of therapeutic agent in the retina choroid tissues of the rabbit eye of between 0.01 ng/mg and 0.50 ng/mg for at least 30, at least 60, at least 90, or at least 120 days after administration of the liquid formulation to the rabbit eyes. In some variations, the liquid formulation when injected between the sclera and conjunctiva of a rabbit eye delivers therapeutic agent giving an average concentration of therapeutic agent in the retina choroid tissues of the rabbit eye of between 0.01 ng/mg and 0.15 ng/mg for at least 30, at least 60, at least 90, or at least 120 days after administration of the liquid formulation to the rabbit eyes. In some variations, the liquid formulation when injected between the sclera and conjunctiva of a rabbit eye delivers therapeutic agent giving an average concentration of therapeutic agent in the retina choroid tissues of the rabbit eye of between 0.01 ng/mg and 0.1 ng/mg for at least 30, at least 60, at least 90, or at least 120_ _ - -----days after administration of the liquid formulation to the rabbit eyes.

[0353] In some variations, the liquid formulation when injected between the sclera and conjunctiva of a rabbit eye delivers tllerapeutic agent giving a ratio of a maximum average concentration of therapeutic agent in the retina choroid tissues of a rabbit eye to a minimum average concentration of therapeutic agent in the retina choroid tissues of a rabbit eye less than 100 for days 30 to at least 60, at least 90, or at least 120 days after administration of the liquid formulation to the rabbit eyes. In some variations, the liquid formulation when injected between the sclera and conjunctiva of a rabbit eye delivers therapeutic agent giving a ratio of a maximum average concentration of therapeutic agent in the retina choroid tissues of a rabbit eye to a minimum average concentration of therapeutic agent in the retina choroid tissues of a rabbit eye less than 50 for days 30 to at least 60, at least 90, or at least 120 days after administration of the liquid formulation to the rabbit eyes. In some variations, the liquid formulation when injected between the sclera and conjunctiva of a rabbit eye delivers therapeutic agent giving a ratio of a maximum average concentration of therapeutic agent in the retina choroid tissues of a rabbit eye to a minimum average concentration of therapeutic agent in the retina choroid tissues of a rabbit eye less than 10 for days 30 to at least 60, at least 90, or at least 120 days after administration of the liquid formulation to the rabbit eyes. In some variations, the liquid formulation when injected between the sclera and conjunctiva of a rabbit eye delivers therapeutic agent giving a ratio of a maximuin average concentration of therapeutic agent in the retina choroid tissues of a rabbit eye to a minimtun average concentration of therapeutic agent in the retina choroid tissues of a rabbit eye less than 5 for days 30 to at least 60, at least 90, or at least 120 days after administration of the liquid formulation to the rabbit eyes.

[0354] In some variations, the liquid forniulation when injected between the sclera and conjunctiva of a rabbit eye delivers therapeutic agent giving an average concentration of therapeutic agent in the retina choroid tissues of a rabbit eye that is approximately constant at a value greater than 0.001 ng/mg for days 30 to at least 60, at least 90, or at least 120 days after administration of the liquid formulation to the rabbit eyes. In some variations, the liquid formulation when injected between the sclera and conjunctiva of a rabbit eye delivers therapeutic agent giving an average concentration of therapeutic agent in the retina choroid tissues of a rabbit eye that is approximately constant at a value.
-------greater than 0.005 ng/mg for days 30 to at least 60, at least 90, or at least 120 days after administration of the liquid formulation to the rabbit eyes. In some variations, the liquid formulation when injected between the sclera and conjunctiva of a rabbit eye delivers therapeutic agent giving an average concentration of therapeutic agent in the retina choroid tissues of a rabbit eye that is approximately constant at a value greater than 0.01 ng/ing for days 30 to at least 60, at least 90, or at least 120 days after administration of the liquid formulation to the rabbit eyes.
[0355] In some variations, the liquid formulation when injected into the vitreous of a rabbit eye delivers therapeutic agent giving an average concentration of therapeutic agent in the vitreous of the rabbit eye of at least 100 ng/mL for at least 30, at least 60, at least 90, or at least 120 days after administration of the liquid fornnulation to the rabbit eyes. In some variations, the liquid formulation when injected into the vitreous of a rabbit eye delivers therapeutic agent giving an average concentration of therapeutic agent in the vitreous of the rabbit eye of at least 1000 ng/mL for at least 30, at least 60, at least 90, or at least 120 days after administration of the liquid formulation to the rabbit eyes. In some variations, the liquid formulation when injected into the vitreous of a rabbit eye delivers therapeutic agent giving an average concentration of therapeutic agent in the vitreous of the rabbit eye of at least 10,000 ng/mL for at least 30, at least 60, at least 90, or at least 120 days after administration of the liquid formulation to the rabbit eyes.
[0356] In some variations, the liquid formulation when injected into the vitreous of a rabbit eye delivers therapeutic agent giving an average concentration of therapeutic agent in the vitreous of the rabbit eye between 100 ng/mL and 100,000 ng/mL for day 30 to at least 60, at least 90, or at least 120 days after administration of the liquid formulation to the rabbit eyes. In some variations, the liquid formulation when injected into the vitreous of a rabbit eye delivers therapeutic agent giving an average concentration of therapeutic agent in the vitreous of the rabbit eye between 100 ng/mL and 50,000 ng/mL for day 30 to at least 60, at least 90, or at least 120 days after administration of the liquid formulation to the rabbit eyes.
[0357] In some variations, the liquid formulation when injected into the vitreous of a rabbit eye delivers therapeutic agent giving an average concentration of therapeutic agent in the vitreous of the rabbit eye between 1000 ng/mL and 100,000 ng/mL for day 30 to at least 60, at least 90, or at least 120 days after administration of the liquid formulation to the rabbit eyes. In some variations, the liquid formulation when injected into the vitreous of a rabbit eye delivers therapeutic agent giving an average concentration of therapeutic agent in the vitreous of the rabbit eye between 1000 ng/mL and 50,000 ng/mL
for day 30 to at least 60, at least 90, or at least 120 days after administration of the liquid formulation to the rabbit eyes.

[03581 In some variations, the liquid formulation when injected into the vitreous of a rabbit eye delivers therapeutic agent giving a ratio of a maximum average concentration of therapeutic agent in the vitreous of the rabbit eye to a minimum average concentration of therapeutic agent in the vitreous of the rabbit eye less than 100 for days 30 to at least 60, at least 90, or at least 120 days after administration of the liquid formulation to the rabbit eyes. In some variations, the liquid formulation when injected into the vitreous of a rabbit eye delivers therapeutic agent giving a ratio of a maximum average concentration of therapeutic agent in the vitreous of the rabbit eye to a minimum average concentration of therapeutic agent in the vitreous of the rabbit eye less than 50 for days 30 to at least 60, at least 90, or at least 120 days after administration of the liquid formulation to the rabbit eyes. In some variations, the liquid formulation when injected into the vitreous of a rabbit eye delivers therapeutic agent giving a ratio of a inaxiinum average concentration of therapeutic agent in the vitreous of the rabbit eye to a ininimum average concentration of therapeutic agent in the vitreous of the rabbit eye less than 10 for days 30 to at least 60, at least 90, or at least 120 days after administration of the liquid formulation to the rabbit eyes.
(0359] In some variations, the liquid formulation when injected into the vitreous of a rabbit eye delivers therapeutic agent giving an average concentration of therapeutic agent in the vitreous of the rabbit eye that is approximately constant at a value greater than 100 ng/mL for days 30 to at least 60, at least 90, or at least 120 days after administration of the liquid formulation to the rabbit eyes. In some variations, the liquid formulation when injected into the vitreous of a rabbit eye delivers therapeutic agent giving an average concentration of therapeutic agent in the vitreous of the rabbit eye that is approximately constant at a value greater than 1000 ng/mL for days 30 to at least 60, at least 90, or at least 120 days after administration of the liquid formulation to the rabbit eyes. In some variations, the liquid formulation when injected into the vitreous of a rabbit eye delivers therapeutic agent giving an average concentration of therapeutic agent in the vitreous of the rabbit eye that is approximately constant at a value greater than 10,000 ng/mL for days 30_to at_least_60, at_1e.ast-90,_or atleast120_days_afteradminis_trationoftheliquid___ formulation to the rabbit eyes.

[0360] In soine variations, the liquid formulation when injected into the vitreous of a rabbit eye delivers therapeutic agent giving an average concentration of therapeutic agent in the retina choroid tissues of the rabbit eye of at least 0.001 ng/mg for at least 30, at least 60, at least 90, or at least 120 days after administration of the liquid formulation to the rabbit eyes. Tn some variations, the liquid formulation wlZen injected into the vitreous of a rabbit eye delivers therapeutic agent giving an average concentration of therapeutic agent in the retina choroid tissues of the rabbit eye of at least 0.01 ng/mg for at least 30, at least 60, at least 90, or at least 120 days after administration of the liquid formulation to the rabbit eyes. In some variations, the liquid fonnulation when injected into the vitreous of a rabbit eye delivers therapeutic agent giving an average concentration of therapeutic agent in the retina choroid tissues of the rabbit eye of at least 0.05 ng/mg for at least 30, at least 60, at least 90, or at least 120 days after administration of the liquid formulation to the rabbit eyes. In some variations, the liquid formulation when injected into tlie vitreous of a rabbit eye delivers therapeutic agent giving an average concentration of therapeutic agent in the retina choroid tissues of the rabbit eye of at least 0.10 ng/mg for at least 30, at least 60, at least 90, or at least 120 days after administration of the liquid formulation to the rabbit eyes.

[0361] In some variations, the liquid formulation when injected into the vitreous of a rabbit eye delivers therapeutic agent giving an average concentration of therapeutic agent in the retina choroid tissues of the rabbit eye between 0.001 ng/mg and 10.00 ng/mg for at least 30, at least 60, at least 90, or at least 120 days after admiziistration of the liquid formulation to the rabbit eyes. In some variations, the liquid formulation when injected into the vitreous of a rabbit eye delivers therapeutic agent giving an average concentration of therapeutic agent in the retina choroid tissues of the rabbit eye between 0.001 ng/mg and 5.00 ng/mg for at least 30, at least 60, at least 90, or at least 120 days after administration of the liquid formulation to the rabbit eyes. 1n some variations, the liquid formulation when injected into the vitreous of a rabbit eye delivers therapeutic agent giving an average concentration of therapeutic agent in the retina choroid tissues of the rabbit eye between 0.001 ng/mg and 1.00 ng/mg for at least 30, at least 60, at least 90, or at least 120 days after administration of the liquid formulation to the rabbit eyes.

[0362] In some variations, the liquid formulation when injected into the vitreous of a-rabbit eye delivers therapeutic agent giving an average concentration of therapeutic agent in the retina choroid tissues of the rabbit eye between 0.01 ng/mg and 10.00 ng/mg for at least 30, at least 60, at least 90, or at least 120 days after administration of the liquid foimulation to the rabbit eyes. In some variations, the liquid formulation when injected into the vitreous of a rabbit eye delivers therapeutic agent giving an average concentration of therapeutic agent in the retina choroid tissues of the rabbit eye between 0.01 ng/mg and 5.00 ng/mg for at least 30, at least 60, at least 90, or at least 120 days after administration of the liquid formulation to the rabbit eyes. In some variations, the liquid formulation when injected into the vitreous of a rabbit eye delivers therapeutic agent giving an average concentration of therapeutic agent in the retina choroid tissues of the rabbit eye between 0.01 ng/mg and 1.00 ng/mg for at least 30, at least 60, at least 90, or at least 120 days after administration of the liqtiid formulation to the rabbit eyes.

[0363] In some variations, the liquid fonnulation when injected into the vitreous of a rabbit eye delivers therapeutic agent giving an average concentration of therapeutic agent in the retina choroid tissues of the rabbit eye between 0.05 ng/mg and 10.00 ng/mg for at least 30, at least 60, at least 90, or at least 120 days after administration of the liquid formulation to the rabbit eyes. In some variations, the liquid fonnulation when injected into the vitreous of a rabbit eye delivers therapeutic agent giving an average concentration of therapeutic agent in the retina choroid tissues of the rabbit eye between 0.05 ng/mg and 5.00 ng/mg for at least 30, at least 60, at least 90, or at least 120 days after administration of the liquid formulation to the rabbit eyes. In some variations, the liquid formulation when injected into the vitreous of a rabbit eye delivers therapeutic agent giving an average concentration of therapeutic agent in the retina choroid tissues of the rabbit eye between 0.05 ng/mg and 1.00 ng/mg for at least 30, at least 60, at least 90, or at least 120 days after administration of the liquid formulation to the rabbit eyes.

[0364] In some variations, the liquid formulation when injected into the vitreous of a rabbit eye delivers therapeutic agent giving an average concentration of therapeutic agent in the retina choroid tissues of the rabbit eye between 0.10 ng/mg and 10.00 ng/mg for at least 30, at least 60, at least 90, or at least 120 days after administration of the liquid formulation to the rabbit eyes. In some variations, the liquid formulation when injected into the vitreous of a rabbit eye delivers therapeutic agent giving an average concentration of therapeutic agent in the retina choroid tissues of the rabbit eye between 0.10 ng/mg and 5.00 ng/mg_for_at_least_30, at least 60,_at_least 90,_or_atleast 120 days afteradministration of the liquid fonnulation to the rabbit eyes. In some variations, the liquid fonnulation when injected into the vitreous of a rabbit eye delivers therapetttic agent giving an average concentration of therapeutic agent in the retina choroid tissues of the rabbit eye between 0.10 ng/mg and 1.00 ng/mg for at least 30, at least 60, at least 90, or at least 120 days after administration of the liquid formulation to the rabbit eyes.

[0365] In some variations, the liquid formulation when injected into the vitreous of a rabbit eye delivers therapeutic agent giving a ratio of a maximum average concentration of therapeutic agent in the retina choroid tissues of the rabbit eye to a minimum average concentration of therapeutic agent in the retina choroid tissues of the rabbit eye less than 100 for days 30 to at least 60, at least 90, or at least 120 days after administration of the liquid formulation to the rabbit eyes. In some variations, the liquid formulation when injected into the vitreous of a rabbit eye delivers therapeutic agent giving a ratio of a maximum average concentration of therapeutic agent in the retina choroid tissues of the rabbit eye to a minimum average concentration of therapeutic agent in the retina choroid tissues of the rabbit eye less than 50 for days 30 to at least 60, at least 90, or at least 120 days after administration of the liquid formulation to the rabbit eyes.

[0366] In some variations, in situ gelling liquid formulations described herein may have in vivo delivery to the retina choroid tissue profiles with the following described characteristics, where the delivery profiles are for delivery of therapeutic agent in vivo after injection of the liquid formulation between the sclera and the conjunctiva of a rabbit eye.

[0367] At day 32 after injection, the percentage in vivo vitreal level may be between about 20% and about 80%, and more usually between about 40% and about 60%, relative to the level present at day 7 after injection. At day 40 after injection, the percentage in vivo vitreal level may be greater than about 20%, and more usually greater than about 40%, relative to the level present at day 7 after injection.

[0368] At day 45 after injection, the percentage in vivo vitreal level may be between about 15% and about 55%, and more usually between about 25% and about 45%, relative to the level present at day 7 after injection. At day 67 after injection, the percentage in vivo vitreal level may be greater thaii about 15%, and more usually greater than about 25%, relative to the level present at day 7 after injection.

[03691 At day 90 after injection, the percentage in vivo vitreal level may be between _about_60%_and_about_100%0, and_more_us.uallybetwe.en_about_70%o._and_about9_0.%0, relative to the level present at day 7 after injection. At day 90 after injection, the percentage in vivo vitreal level may be greater than about 60%, and more usually greater than about 70%, relative to the level present at day 7 after injection.

[0370] In some variations, the percentage in vivo vitreal level has the following characteristics relative to the level present at day 7 after injection: at 32 days after injection it is less than about 80%; at 45 days after injection it is less than about 60%; and 90 days after injection it is less than about 100%.

[0371] In some variations, the liquid formulation wllen injected between the sclera and conjunctiva of a rabbit eye delivers therapeutic agent giving an average concentration of therapeutic agent in the retina choroid tissues of the rabbit eye of at least about 0.1 pg/mg for at least about 30, at least about 60, or at least about 90 days after administration of the liquid formulation to the rabbit eye. In some variations, the liquid formulation when injected between the sclera and conjunctiva of a rabbit eye delivers therapeutic agent giving an average concentration of therapeutic agent in the vitreous of the rabbit eye of at least about 0.01 ng/mg for at least about 30, at least about 60, or at least about 90 days after administration of the liquid formulation to the rabbit eye. In some variations, the liquid formulation when injected between the sclera and conjunctiva of a rabbit eye delivers therapeutic agent giving an average concentration of therapeutic agent in the vitreous of the rabbit eye of at least about 0.1 ng/mg for at least about 30, at least about 60, or at least about 90 days after administration of the liquid formulation to the rabbit eye. In some variations, the liquid formulation when injected between the sclera and conjunctiva of a rabbit eye delivers therapeutic agent giving an average concentration of therapeutic agent in the vitreous of the rabbit eye of at least about 1 ng/mL
for at least about 30, at least about 60, or at least about 90 days after administration of the liquid formulation to the rabbit eye.

[0372] In some variations, the ratio of the base ten logarithms of the average levels of a therapeutic agent in two or more of the retina choroid tissues, the sclera, and the vitreous is approximately constant over an extended period of time after placement of the in situ gelling formulation in or proximate to the eye. In some variations, the ratio of the base ten logarithms of the average levels of a therapeutic agent in two or more of the retina choroid tissues, the sclera, and the vitreous is approximately constant over an extended period_oftime after_placement of the_in situ gellingformulatiorLbetween the_s_clera_and the_ conjunctiva of an eye. In some variations, the ratio of the base ten logarithms of the average levels of a therapeutic agent in the vitreous and the sclera is approximately constant over an extended period of time after placement of the in situ gelling formulation between the sclera and the conjunctiva of an eye.

[0373] In some variations, the ratio of the base ten logarithms of the average levels of a therapeutic agent in the vitreous and the retina choroid tissues is approximately constant over an extended period of time. Put another way, as the level of therapeutic agent in the vitreous rises, the level of therapeutic agent in the retina choroid tissues rises to a similar degree when considered on the logarithmic scale, and vice versa.

[0374] In some variations, the ratio of the base ten logarithms of the average levels of a therapeutic agent in the vitreous versus the retina choroid tissues is approximately constant over an extended period of time of about 7, about 30, about 60, or about 90 days.
In some variations, the ratio of the average level of therapeutic agent in the vitreous relative to the level of therapeutic agent in the retina choroid tissues after placement of the in situ gelling formulation between the sclera and the conjunctiva of an eye is constant at about 37:1 at day 7, about 40:1 at day 32, about 10:1 at day 45, and about 34:1 at day 90.
[0375] In some variations, the ratio of the average level of therapeutic agent in the vitreous relative to the level of therapeutic agent in the retina choroid tissues is constant at about 40:1 over a period of about 7, about 32, about 45, or about 90 days.

[0376] In some variations, the average level of the therapeutic agent in any or all of the retina choroid tissues, the sclera, and the vitreous is approximately constant over an extended period of time after placement of the in situ gelling formulation in or proximate to the eye.

[0377] In some variations, after placement of an in situ gelling formulation between the sclera and the conjunctiva, the average level of therapeutic agent in the vitreous is approximately constant at about 8.1 ng/ml. In some variations, after placement of an in situ gelling formulation between the sclera and the conjunctiva, the average level of therapeutic agent in the retina choroid tissues is approximately constant at about 0.25 ng/mg. In some variations, after placement of an in situ gelling formulation between the sclera and the conjunctiva, the average level of therapeutic agent in the sclera is approximately constant at about 1930 ng/mg.

[0378] In some variations, the in situ gelling formulation when injected between the __ _sclera and conjunctiva_of a..rabbit _e.y_esnaintains__an.a_verage_level of therapeutic_agent in the vitreous that is approximately constant at about 0.1 pg/mL for at least about 30, at least about 60, or at least about 90 days after administration of the liquid formulation to the rabbit eye. In some variations, the in situ gelling formulation when injected between the sclera and conjunctiva of a rabbit eye maintains an average level of therapeutic agent in the vitreous that is approximately constant at about 0.001 ng/mL for at least about 30, at least about 60, or at least about 90 days after administration of the liquid formulation to the rabbit eye. In some variations, the in situ gelling formulation when injected between the sclera and conjunctiva of a rabbit eye maintains an average level of therapeutic agent in the vitreous that is approximately constant at about 0.01 ng/mL for at least about 30, at least about 60, or at least about 90 days after administration of the liquid formulation to the rabbit eye. In some variations, the in situ gelling formulation when injected between the sclera and conjunctiva of a rabbit eye maintains an average level of therapeutic agent in the vitreous that is approximately constant at about 0.1 ng/mL for at least about 30, at least about 60, or at least about 90 days after administration of the liquid formulation to the rabbit eye. In some variations, the in situ gelling formulation when injected between the sclera and conjunctiva of a rabbit eye maintains an average level of therapeutic agent in the vitreous that is approximately constant at about 1 ng/mL for at least about 30, at least about 60, or at least about 90 days after administration of the liquid formulation to the rabbit eye. In some variations, the in situ gelling forrnulation when injected between the sciera and conjunctiva of a rabbit eye maintains an average level of therapeutic agent in the vitreous that is approximately constant at about 10 ng/mL for at least about 30, at least about 60, or at least about 90 days after administration of the liquid formulation to the rabbit eye. In some variations, the in situ gelling formulation when injected between the sclera and conjunctiva of a rabbit eye maintains an average level of therapeutic agent in the vitreous that is approximately constant at about 100 ng/mL for at least about 30, at least about 60, or at least about 90 days after administration of the liquid formulation to the rabbit eye.

[03791 In some variations, the in situ gelling formulation when injected between the sclera and conjunctiva of a rabbit eye maintains an average level of therapeutic agent in the retina choroid tissues that is approximately constant at about 0.1 pg/mg for at least about 30, at least about 60, or at least about 90 days after administration of the liquid formulation to the rabbit eye. In some variations, the in situ gelling formulation when injected between the sclera and conjunctiva of a rabbit eye maintains an average level therapeutic agent in the retina choroid tissues that is approximately constant at about 0.001 ng/mg for at least about 30, at least about 60, or at least about 90 days after administration of the liquid formulation to the rabbit eye. In some variations, the in situ gelling formulation when injected between the sclera aild conjunctiva of a rabbit eye maintains an average level of therapeutic agent in the retina choroid tissues that is approximately constant at about 0.01 ng/mg for at least about 30, at least about 60, or at least about 90 days after administration of the liquid formulation to the rabbit eye. In some variations, the in situ gelling formulation when injected between the sclera and conjunctiva of a rabbit eye maintains an average level of therapeutic agent in the retina choroid tissues that is approximately constant at about 0.1 ng/mg for at least about 30, at least about 60, or at least about 90 days after administration of the liquid formulation to the rabbit eye. In some variations, the in situ gelling formulation wlien injected between the sclera and conjunctiva of a rabbit eye maintains an average level of therapeutic agent in the retina choroid tissues that is approximately constant at about 1 ng/mg for at least about 30, at least about 60, or at least about 90 days after administration of the liquid formulation to the rabbit eye. In some variations, the in situ gelling formulation when injected between the sclera and conjunctiva of a rabbit eye maintains an average level of therapeutic agent in the retina choroid tissues that is approximately constant at about 10 ng/mg for at least about 30, at least about 60, or at least about 90 days after administration of the liquid formulation to the rabbit eye.

[0380] In some variations, the in situ gelling formulation when injected between the sclera and conjunctiva of a rabbit eye maintains an average level of therapeutic agent in the sclera that is approximately constant at about 0.1 pg/mg for at least about 30, at least about 60, or at least about 90 days after administration of the liquid formulation to the rabbit eye. In some variations, the in situ gelling formulation when injected between the sclera and conjunctiva of a rabbit eye maintains an average level of therapeutic agent in the sclera that is approximately constant at about 0.001 ng/mg for at least about 30, at least about 60, or at least about 90 days after administration of the liquid formulation to the rabbit eye. In some variations, the in situ gelling formulation when injected between the sclera and conjunctiva of a rabbit eye maintains an average level of therapeutic agent in the sclera that is approximately constant at about 0.01 ng/mg for at least about 30, at least about 60, or at least about 90 days after administration of the liquid formulation to the rabbit eye. In some variations, the in situ gelling formulation when injected between - - - - - ------the sclera and conjunctiva of a rabbit eye maintains an average level of therapeutic agent in the sclera that is approximately constant at about 0.1 ng/mg for at least about 30, at least about 60, or at least about 90 days after administration of the liquid formulation to the rabbit eye. In some variations, the in situ gelling foiTnulation when injected between the sclera and conjunctiva of a rabbit eye maintains an average level of therapeutic agent in the sclera that is approximately constant at about 1 ng/mg for at least about 30, at least about 60, or at least about 90 days after administration of the liquid formulation to the rabbit eye. In some variations, the in situ gelling fonnulation when injected between the sclera and conjunctiva of a rabbit eye maintains an average level of therapeutic agent in the sclera that is approximately constant at about 10 ng/mg for at least about 30, at least about 60, or at least about 90 days after administration of the liquid formulation to the rabbit eye. In some variations, the in situ gelling forn.iulation when injected between the sclera and conjunctiva of a rabbit eye maintains an average level of therapeutic agent in the sclera that is approximately constant at about 100 ng/mg for at least about 30, at least about 60, or at least about 90 days after administration of the liquid formulation to the rabbit eye. In some variations, the in situ gelling formulation when injected between the sciera and coiijunctiva of a rabbit eye maintains an average level of therapeutic agent in the sclera that is approximately constant at about 1 g/mg for at least about 30, at least about 60, or at least about 90 days after administration of the liquid formulation to the rabbit eye. In some variations, the in situ gelling formulation when injected between the sclera and conjunctiva of a rabbit eye maintains an average level of therapeutic agent in the sclera that is approximately constant at about 10 g/mg for at least about 30, at least about 60, or at least about 90 days after administration of the liquid formulation to the rabbit eye.

[0381] For treatment, prevention, inhibition, delaying the onset of, or causing the regression of certain diseases or conditions, it may be desirable to maintain delivery of a therapeutically effective amount of the therapeutic agent for an extended period of time.
Depending on the disease or condition being treated, prevented, inhibited, having onset delayed, or being caused to regress this extended period of time may be at least about 1 week, at least about 2 weeks, at least about 3 weeks, at least about 1 month, at least about 3 months, at least about 6 months, at least about 9 months, or at least about 1 year.
Generally, however, any extended period of delivery may be possible. A
therapeutically effective amount of agent may be delivered for an extended period by a liquid formulatiou or composition that maintains for the extended period a concentration of agent in a subject or an eye of a subject sufficient to deliver a therapeutically effective amount of agent for the extended time.

[03821 Delivery of a therapeutically effective amount of the therapeutic agent for an extended period may be achieved via placement of one composition or liquid formulation or may be achieved by application of two or more doses of composition or liquid formulations. As a non-limiting example of such multiple applications, maintenance of the therapeutic amount of rapamycin for 3 months for treatment, prevention, inhibition, delay of onset, or cause of regression of wet AMD may be achieved by application of one liquid formulation or composition delivering a therapeutic amount for 3 months or by sequential application of a plurality of liquid formulations or compositions.
The optimal dosage regime will depend on the therapeutic amount of the therapeutic agent needing to be delivered, and the period over which it need be delivered. Those versed in such extended therapeutic agent delivery dosing will understand how to identify dosing regimes that may be used based on the teachings provided herein.

[0383] When using certain therapeutic agents or for the treatment, prevention, inhibition, delaying the onset of, or causing the regression of certain diseases, it may be desirable for delivery of the therapeutic agent not to commence immediately upon placement of the liquid fonnulation or composition into the eye region, but for delivery to commence after some delay. For example, but in no way limiting, such delayed release may be useful where the therapeutic agent inhibits or delays wound healing and delayed release is desirable to allow healing of any wounds occurring upon placement of the liquid formulation or composition. Depending on the therapeutic agent being delivered and/or the diseases and conditions being treated, prevented, inhibited, onset delayed, and regression caused this period of delay before delivery of the therapeutic agent commences may be about 1 hour, about 6 hours, about 12 hours, about 18 hours, about 1 day, about 2 days, about 3 days, about 4 days, about 5 days, about 6 days, about 7 days, about 8 days, about 9 days, about 10 days, about 11 days, about 12 days, about 13 days, about 14 days, about 21 days, about 28 days, about 35 days, or about 42 days. Other delay periods may be possible. Delayed release formulations that may be used are known to people versed in the technology.

Intvavitreal and subconjunctival delivery of s=apanzycin for treatsnent, ps=evention, inhibition, delay of onset, or cause of regression of AMD

[0384] In one method described herein, a liquid formulation comprising rapamycin is delivered subconjunctivally or to the vitreous of an eye to prevent, treat, inhibit, delay onset of, or cause regression of angiogenesis in the eye, including but not limited to treating CNV as observed, for example, in AMD. In some va.riations, the liquid formulation is used to treat angiogenesis in the eye, including but not limited to treating CNV as obseived, for example, in AMD. Rapamycin has been shown to inliibit CNV
in rat and mice models, as described in U.S. Application No. 10/665,203, which is incorporated herein by reference in its entirety. Rapamycin has been observed to inhibit MatrigelTM and laser-induced CNV when administered systemically and subretinally.
Also, periocular injection of rapamycin inhibits laser-induced CNV.

[0385) Other therapeutic agents that may be delivered to the eye, particularly the vitreous of an eye, for treatment, prevention, inhibition, delaying onset, or causing regression of angiogenesis in the eye (such as CNV) are members of the liinus family of compounds other than rapamycin including but not limited to everolimus and tacrolimus (FK-506).

[0386] As described herein, the dosage of the therapeutic agent will depend on the condition being addressed, whether the condition is to be treated, prevented, inhibited, have onset delayed, or be caused to regress, the particular therapeutic agent, and other clinical factors such as weight and condition of the subject and the route of administration of the therapeutic agent. It is to be understood that the methods, liquid formulations, and compositions described herein have application for both human and veterinary use, as well as uses in other possible animals. As described herein, tissue concentrations of therapeutic agents expressed in units of mass per volume generally refer to tissues that are primarily aqueous such as the vitreous, for example. Tissue concentrations of therapeutic agents expressed in unit of mass per mass generally refer to other tissues such as the sclera or retina choroid tissues, for example.

[03871 One concentration of rapamycin that may be used in the methods described herein is one that provides about 0.01 pg/ml or pg/mg or more of rapamycin at the tissue level. Another concentration that may be used is one that provides about 0.1 pg/ml or ng/mg or more at the tissue level. Another concentration that may be used is one that provides about 1 pg/ml or ng/mg or more at the tissue level. Another concentration_that may be used is one that provides about 0.01 ng/ml or ng/mg or more at the tissue level.
Another concentration that may be used is one that provides about 0.1 ng/ml or ng/mg or more at the tissue level. Another concentration that may be used is one that provides about 0.5 ng/inl or ng/mg or more at the tissue level. Another concentration that may be used is one that provides about 1 ng/ml or more at the tissue level. Another concentration that may be used is one that provides about 2 ng/ml or more at the tissue level. Another concentration that may be used is one that provides about 3 ng/nil or more at the tissue level. Another concentration that may be used is one that provides about 5 ng/ml or more at the tissue level. Another concentration that may be used is one that provides about 10 ng/ml or more at the tissue level. Another concentration that may be used is one that provides about 15 nghnl or more at the tissue level. Another concentration that may be used is one that provides about 20 ng/ml or more at the tissue level. Another concentration that may be used is one that provides about 30 ng/ml or more at the tissue level. Another concentration that may be used is one that provides about 50 ng/ml or more at the tissue level. One of ordinary skill in the art would know how to arrive at an appropriate concentration depending on the route arid duration of administration utilized, given the teachings herein.

[0388] Generally, the amount of rapamycin administered in a liquid formulation is an amount sufficient to treat, prevent, inhibit, delay the onset, or cause regression of the disease or condition of the eye for the required amount of time. In some variations the amount of rapamycin administered in the liquid formulation is an amount sufficient to treat the disease or condition of the eye for the required amount of time.

[0389] In some variations, a total amount of rapamycin less than about 5 mg is administered subconjunctivally. In some variations, a total amount of rapamycin less than about 5.0 mg is administered subconjunctivally. In some variations, a total amount of rapamycin less than about 4.5 mg is administered subconjunctivally. In some variations, a total amount of rapamycin less than about 4.0 mg is administered subconjunctivally. In some variations, a total amount of rapamycin less than about 3.5 ing is administered subconjunctivally. In some variations, a total anlount of rapainycin less than about 3.0 mg is administered subconjunctivally. In some variations, a total amount of rapamycin less than about 2.5 mg is administered subconjunctivally. In some variations, a total amount of rapamycin less than about 2 mg is administered subconjunctivally. In some variations, a total amount of rapamycin less than about 1.2 mg is administered subconjunctivally._In some variations, a total amount of rapamycin less than about 1.0 mg is administered subconjunctivally. In some variations, a total amount of rapamycin less than about 0.8 mg is administered subconjunctivally. In some variations, a total amount of rapamycin less than about 0.6 mg is administered sttbconjunctivally. In some variations, a total ainount of rapamycin less than about 0.4 mg is administered subconjunctivally. In some variations, a volume of a formulation is administered that contains an amount of rapamycin described herein.

[0390] In some variations, a liquid formulation containing a concentration of rapamycin by weight of the total of between abotit 0.5% and about 6% is subconjunctivally administered to a human subject by administering between about 0.1 l and about 200 l of a liquid formulation described herein. In some variations, a liquid formulation containing a concentration of rapamycin by weight of the total of between about 0.5% and about 4% is subconjunctivally administered to a human subject by administering between about 1 l and about 50 ,ul of a liquid formulation described herein. In some variations, a liquid formulation containing a concentration of rapamycin by weight of the total of between about 1.5% and about 3.5% is subconjunctivally administered to a huinan subject by administering between about 1 l and about 15 l of a liquid formulation described herein. In some variations, a liquid formulation containing a concentration of rapamycin by weight of the total of about 2% is subconjunctivally administered to a human subject by administering between about 1 l and about 15 l of a liquid formulation described herein.

[0391] In some variations, a liquid formulation containing an amount of rapamycin of between about 0.2 g and about 4 mg is subconjunctivally administered to a human subject by administering between about 0.1 l and about 200 .l of a liquid formulation described herein. In some variations, a liquid formulation containing an amount of rapamycin of between about 20 g and about 2 mg is subconjunctivally administered to a human subject by administering between about 1 l and about 100 ,ul of a liquid formulation described herein. In some variations, a liquid formulation containing an amount of rapamycin of between about 20 g and about 1 mg is subconjunctivally administered to a htunan subject by administering between about 1 l and about 50 l of a liquid formulation described herein.In some variations, a liquid formulation containing an amount of rapamycin of between about 20 g and about 500 g is subconjunctivally administered to a human subj ect by__administering_ between about 1 l_and about 25_Al_ of a_ liquid formulation described herein. In some variations, a liquid formulation containing an amount of rapamycin of between about 20 g and about 300 g is subconjunctivally administered to a human subject by administering between about I l and about 15 l of a liquid fozmulation described herein.
[0392] Iti some variations, a total amount of rapamycin less than about 200 g is administered intravitreally. In some variations, a total amount of rapamycin less than about 200 ~tg is administered intravitreally. In some variations, a total ainount of rapaiiaycin less tlzan about 300 g is administered intravitreally. In some variations, a total amount of rapamycin less than about 400 g is administered intravitreally. In some variations, a total amount of rapamycin less than about 500 ~tg is administered intravitreally. In some variations, a total amount of rapamycin less than about 600 g is administered intravitreally. In some variations, a total amount of rapamycin less than about 800 ttg is administered intravitreally. In some variations, a total amount of rapamycin less than about 1 mg is adininistered intravitreally. In some variations, a total amount of rapamycin less than about 2 mg is administered intravitreally. In some variations, a total amount of rapamycin less than about 2.5 mg is adininistered intravitreally. In some variations, a total amount of rapamycin less than about 3 mg is administered intravitreally. In some variations, a total amount of rapaniycin less than about 3.5 mg is administered intravitreally. In some variations, a total amount of rapamycin less than about 4 mg is administered intravitreally. In some variations, a volume of a fonnulation is administered that contains an amount of rapamycin described herein.
[03931 In some variations, a liquid formulation containing a concentration of rapamycin by weight of the total of between about 0.5% and about 6% is intravitreally administered to a human subject by administering between about 0.1 l and about 200 ,ul of a liquid fomlulation described herein. In some variations, a liquid formulation containing a concentration of rapamycin by weight of the total of between about 0.5% and about 4% is intravitreally administered to a human subject by administering between about I l and about 50 l of a liquid formulation described herein. In some variations, a liquid formulation containing a concentration of rapamycin by weight of the total of between about 1.5% and about 3.5% is intravitreally administered to a human subject by administering between about 1,ul and about 15 l of a liquid formulation described herein. In some variations, a liquid formulation containing a concentration of rapalnycin- __---by weight of the total of about 2% is intravitreally administered to a human subject by administering between about 1,cd and about 15 l of a liquid formulation described herein.

[0394] In some variations, a liquid foimulation containing an aznount of rapanlycin of between about 0.2 g and about 4 mg is intravitreally administered to a human subject by administering between about 0.1 l and about 200 l of a liquid formulation described herein. In some variations, a liquid formulation containing an atnount of rapamycin of between about 20 g and about 2 ing is intravitreally administered to a human subject by administering between about 1 l and about 100 l of a liquid formulation described herein. In some variations, a liquid formulation containing an amount of rapamycin of between about 20 .g and about 1 mg is intravitreally administered to a huinan subject by administering between about 1 l and about 50 l of a liquid formulation described hereiii. In some variations, a liquid formulation containing an amount of rapamycin of between about 20 g and about 500 g is intravitreally administered to a human subject by administering between about 1 l and about 25 l of a liquid formulation described herein. In some variations, a liquid formulation containing an amount of rapamycin of between about 20 g and about 300 g is intravitreally administered to a human subject by administering between about 1 l and about 15 l of a liquid formulation described herein.

[0395] In some variations a liquid formulation as described herein containing an amount of rapamycin of between about 1 gg and about 5 mg is adniinistered to a human subject for treatment of wet AMD. In some variations a liquid formulation as described herein containing an amount of rapamycin of between about 20 gg and about 4 mg is administered to a human subject for treatment of wet AMD. In some variations a liquid formulation as described herein containing an amount of rapamycin of between about 20 gg and about 1.2 mg is administered to a human subject for treatment of wet AMD. In some variations an amount of rapamycin of between about 10 g and about 0.5 mg is administered to a human subject for treatment of wet AMD. In some variations an amount of rapamycin of between about 10 gg and 90 g is administered to a human subject for treatment of wet AMD. In some variations an amount of rapamycin of between about 60 g and about 120 .g is administered to a human subject for treatment of wet AMD. In some variations an amount of rapamycin of between about 100 g_and about 400 g is administered to a human subject for treatment of wet AMD. In some variations an amount ofrapamycin of between about 400 g and about 1 mg is administered to a huma.n subject for treatment of wet AMD. In some variations an amount of rapamycin of between about 1 mg and about 5 mg is administered to a human subject for treatment of wet AMD. In some variations, an amount of rapamycin of between about 3 mg and about 7 mg is administered to a human subject for treatment of wet AMD. In some variations, an amount of rapamycin of between about 5 mg and about mg is administered to a human subject for treatment of wet AMD.

[0396] In some variations a liquid formulation as described herein containing an amount of rapamycin of between about 1 gg and about 5 mg is administered to a human subject for prevention of wet AMD. In some variations a liquid formulation as described herein containing an amount of rapamycin of between about 20 gg and about 4 mg is administered to a human subject for prevention of wet AMD. In some variations a liquid formulation as described herein containing an amount of rapamycin of between about 20 g and about 1.2 mg is administered to a human subject for prevention of wet AMD. In some variations an amount of rapamycin of between about 10 g and about 0.5 mg is administered to a human subject for prevention of wet AMD. In some variations an amount of rapamycin of between about 10 gg and 90 g is administered to a human subject for prevention of wet AMD. In some variations an amount of rapamycin of between about 60 g and about 120 g is administered to a human subject for prevention of wet AMD. In some variations an amount of rapamycin of between about 100 g and about 400 g is administered to a human subject for prevention of wet AMD. In some variations an amount of rapamycin of between about 400 gg and about 1 mg is administered to a human subject for prevention of wet AMD. In some variations an amount of rapamycin of between about 1 mg and about 5 mg is administered to a human subject for prevention of wet AMD. In some variations, an amount of rapamycin of between about 3 mg and about 7 mg is administered to a huinan subject for prevention of wet AMD. In some variations, an amount of rapamycin of between about 5 mg and about 10 mg is administered to a human subject for prevention of wet AMD.

[0397] In some variations a liquid formulation as described herein containing an amount of rapamycin of between about 1 .g and about 5 mg is administered to a human subject for treatment of dry AMD. In some variations a liquid formulation as described herein containing an amount of rapamycin of between about 20 gg and about 4mg_is _ ---administered to a human subject for treatment of dry AMD. In some variations a liquid formulation as described herein containing an amount of rapamycin of between about 20 gg and about 1.2 mg is administered to a human subject for treatment of dry AMD. In some variations an amount of rapamycin of between about 10 gg and about 0.5 mg is administered to a human subject for treatment of dry AMD. In some variations an amount of rapamycin of between about 10 gg and 90 gg is administered to a human subject for treatment of dry AMD. In some variations an amount of rapamycin of between about 60 gg and about 120 .g is administered to a human subject for treatment of dry AMD. In some variations an amount of rapamycin of between about 100 gg and about 400 g is administered to a human subject for treatment of dry AMD. In some variations an amount of rapamycin of between about 400 g and about 1 mg is administered to a human subject for treatment of dry AMD. In some variations an amount of rapamycin of between about 1 mg and about 5 mg is administered to a human subject for treatment of dry AMD. In some variations, an amount of rapamycin of between about 3 mg and about 7 mg is administered to a human subject for treatment of dry AMD. In some variations, an amount of rapamycin of between about 5 mg and about 10 mg is administered to a huinan subject for treatment of dry AMD.

[0398] In some variations, a liquid formulation as described herein containing an amount of rapamycin of between about 1 g and about 5 mg is administered to a human subject for treatment of angiogenesis, including but not limited to choroidal neovascularization. In some variations, an amount of rapamycin of between about 20 gg and about 4 mg is administered to the human subject; between about 20 g and about 1.2 mg; between about 10 g and about 0.5 mg is adnlinistered to a human subject for treatment of wet AMD, between about 10 g and 90 g, between about 60 g and 120 g is administered to the human subject; between about 100 g and 400 g, between about 400 gg and 1 mg is administered to the human subject; in some variations, an amount of rapamycin of between about 1 mg and 5 mg is administered to the human subject ; in some variations, an amount of rapamycin of between about 3 mg and 7 mg is administered to the human subject; in some variations, an amount of rapamycin of between about 5 mg and 10 mg is administered to the human subject for treatment of angiogenesis, including but not limited to choroidal neovascularization.

[0399] In one method, a liquid formulation as described herein contains an amount of a therapeutic agent equivalent to an amount of rapamycin.

[0400] In one method, a liquid formulation as described herein containing an amount of a therapeutic agent equivalent to an amount of rapamycin of between about 1 g and about 5 mg is administered to a human subject for treatment of wet AMD. In some variations, an amount of a therapeutic agent equivalent to an amount of rapamycin of between about 1 g and about 5 mg is administered to the human subject;
between about 20 g and about 1.2 mg; between about 10 g and about 0.5 mg is administered to a liuman subject for treatment of wet AMD, between about 10 g and 90 g, between about 60 g and 120 g is administered to the human subject; between about 100 g and 400 g, between about 400 g and 1 mg is administered to the human subject is administered to the human subject; in some variations, an amount of a therapeutic agent equivalent to an amount of rapamycin of between about 1 ing and 5 mg is administered to the human subject; in some variations, an amount of a therapeutic agent equivalent to an amount of rapamycin of between about 3 mg and 7 mg is administered to the human subject;
in some variations, an amount of a therapeutic agent equivalent to an amount of rapamycin of between about 5 mg and 10 ing is administered to the human subject.

[0401] In some variations, a liquid formulation as described herein containing an amount of a therapeutic agent equivalent to an amount of rapamycin of between about 1 g and about 5 mg is administered to a human subject for treatment of dry AMD.
In some variations, an amount of a therapeutic agent equivalent to an amount of rapamycin of between about 20 g and about 4 mg is administered to the human subject;
between about 20 gg and about 1.2 mg; between about 10 g and about 0.5 mg is administered to a human subject for treatment of wet AMD, between about 10 g and 90 g, between about 60 gg and 120 g is administered to the huinan subject; between about 100 g and 400 g, between about 400 g and 1 mg is administered to the human subject; in some variations, an amount of a therapeutic agent equivalent to an amount of rapamycin of between about 400 g and 1 mg is administered to the human subject; in some variations, an amount of a therapeutic agent equivalent to an amount of rapamycin of between about 1 mg and 5 mg is administered to the human subject; in some variations, an amount of a therapeutic agent equivalent to an amount of rapamycin of between about 3 mg and 7 mg is administered to the human subject; in some variations, an amount of a therapeutic agent equivalent to an amount of rapamycin of between about 5 mg and 10 mg is administered to the human subject to treat dry AMD.

[0402] In some variations, a liquid formulation as described herein containing_an amount of a therapeutic agent equivalent to an amount of rapamycin of between about 1 g and about 5 mg is administered to a human subject for prevention of wet AMD.
In some variations, an amount of a therapeutic agent equivalent to an amount of rapamycin of between about 20 g and about 4 mg is administered to the huinan subject;
between about 20 g and about 1.2 mg; between about 10 g and about 0.5 mg is administered to a human subject for prevention of wet AMD, between about 10 g and 90 g, between about 60 g and 120 g is administered to the human subject; between about 100 g and 400 g, between about 400 g and 1 ing is administered to the human subject;
in some variations, an amount of a therapeutic agent equivalent to an amount of rapamycin of between about 400 g and 1 mg is administered to the human subject; in some variations, an amount of a therapeutic agent equivalent to an amount of rapamycin of between about 1 ing and 5 mg is administered to the human subject; in some variations, an amount of a therapeutic agent equivalent to an amount of rapamycin of between about 3 mg and 7 mg is administered to the human subject; in some variations, an amount of a tlierapeutic agent equivalent to an amount of rapamycin of between about 5 mg and 10 mg is adininistered to the human subject to prevent wet AMD.

[0403] In some variations, any one or more of the formulations described herein are administered intravitreally every 3 or more months, every 6 or inore months, every 9 or more months, or every 12 or more months, or longer, to treat one or more of choroidal neovascularization, wet AMD, dry AMD, to prevent wet AMD, or to prevent progression of dry AMD to wet AMD. In some variations, any one or more of the formulations described herein are administered subconjunctivally every 3 or more months, every 6 or more months, every 9 or more months, or every 12 or more months, or longer, to treat one or more of choroidal neovascularization, wet AMD, dry AMD, or to prevent wet AMD.
[0404] In some variations, any one or more of the rapamycin formulations described herein are administered intravitreally every 3 or more months, every 6 or more months, every 9 or more months, or every 12 or more months, or longer, to treat one or more of choroidal neovascularization, wet AMD, dry AMD, to prevent wet AMD, or to prevent progression of dry AMD to wet AMD. In some variations, any one or more of the rapamycin formulations described herein are administered subconjunctivally every 3 or more months, every 6 or more months, every 9 or more months, or every 12 or more months, or longer, to treat one or more of choroidal neovascularization, wet AMD, dry AMD, or to prevent wet AMD. In some variations, the effect of the rapamy_cin persists beyond the period during which it is present in the ocular tissues.

[0405] Delivery of the therapeutic agents described herein may, for example, be delivered at a dosage range between about 1 ng/day and about 100 g/day, or at dosages higher or lower than this range, depending on the route and duration of administration. In some variations of liquid formulation or composition used in the methods described herein, the therapelrtic agents are delivered at a dosage range of between about 0.1 g/day and about 10 g/day. In some variations of liquid forinulation or composition used in the methods described herein, the therapeutic agents are delivered at a dosage range of between about 1 g/day and about 5 g/day. Dosages of various therapeutic agents for treatment, prevention, inhibition, delay of onset, or cause of regression of various diseases and conditions described herein can be refined by the use of clinical trials.

[0406] The liquid formulations, including but not limited to solutions, suspensions, emulsions and situ gelling formulations, and coinpositions described herein maybe used for delivery to the eye, as one nonlimiting example by ocular or periocular administration, of therapeutically effective amounts of rapainycin for extended periods of time to treat, prevent, inhibit, delay the onset of, or cause regression of CNV, and thus may be used to treat, prevent, inhibit, delay the onset of, or cause regression of wet AMD, or transition of dry AMD to wet AMD. It is believed that by changing certain characteristics of the liquid formulations described herein, including but not limited to the components of the liquid formulations, the location in the eye to which the liquid formulation is delivered, including without limitation subconjunctival or intravitreal placement, the liquid formulations may be used to deliver therapeutically effective amounts of rapamycin to the eye for a variety of extended time periods including delivery of therapeutic amounts for greater than about 1 week, for greater than about 2 weeks, for greater than about 3 weeks, for greater than about 1 month, for greater than about 3 months, for greater than about 6 months, for greater than about 9 months, for greater than about 1 year.

[0407] When a therapeutically effective amount of rapamycin is administered to a subject suffering from wet AMD, the rapamycin may treat, inhibit, or cause regression of the wet AMD. Different therapeutically effective amounts may be required for treatment, inhibition or causing regression. A subject suffering from wet AMD may have CNV
lesions, and it is believed that administration of a therapeutically effective amount of rapamycin may have a variety of effects, including but not limited to causing regression of the CNV lesions, stabilizing the CNV lesion, and preventing progression of an active CNV lesion.

[0408] When a therapeutically effective amount of rapamycin is administered to a subject suffering from dry AMD, it is believed that the rapamycin may prevent or slow the progression of dry AMD to wet AMD.

EXAMPLES
[0409] Unless the context indicates otherwise, the error bars in the charts show one standard deviation. Where ethanol is used, it is 200 proof ethanol from Gold Shield Distributors, Hayward, CA. Where rapamycin is used, it is from LC
laboratories, Woburn, MA, or Chunghwa Chemical Synthesis & Biotech Co., LTD (CCSB), Taipei Hsien, Taiwan, ROC. Where PEG 400 is used, it is from The Dow Chemical Company, New Milford, CT. Some of the graphs use the expression "uL" or "ug" to refer to L
or ,ug, respectively. Where a volume of 10 L or less is administered, Hamilton HPLC
syringes were used.

Exanzple 1- Prepaf-ation and Chaf-acteNization of a Rapanaycin-Containing Solution [0410] 1.256% rapamycin (percentage of the total weight) was dissolved in 9.676%
ethanol (percentage of the total weight). An aqueous solution of 15% F127 (Lutrol) in sterile water was slowly added under continuous agitation. The final concentration was approximately 78.57% sterile water (percentage of the total weight) and approximately 10.50% F127 (Lutrol) (percentage of the total weight). This solution is listed as formulation #32 in Table 1. The solution was placed at 2 C until use.

Exan2ple 2 - Subconjunctival If jection of a Rapainycin-Containing Solution [0411] 50 gl of the solution described in Example 1 was injected between the sclera and the conjunctiva of the eye of New Zealand white rabbits.

[0412] Fig. 2 depicts the average concentration of rapamycin present in the vitreous (ng/ml), retina choroid (ng/mg), and sclera (ng/mg) on a logarithmic scale at 20, 40, 67, and 90 days after injection.

[0413] The analysis was by liquid chromatography mass spectroscopy (LCMS) using an internal standard.

[0414] At each timepoint, the average concentration of rapamy_cin was calculate_d_by __-_ adding the concentrations of rapamycin obtained for each eye from each rabbit, and dividing the total by the number of eyes analyzed. In this experiment, each timepoint represents the average of either two eyes of each of two rabbits (four eyes at that timepoint) or the average of two eyes of one rabbits (two eyes at that timepoint).

[04151 The full vitreous was homogenized and analyzed. The average concentration of the vitreous was calculated by dividing the mass of rapamycin measured by the volume of vitreous analyzed. The sample did not include the site of administration;
tlnts, this measurement indicated the level of z-apamycin delivered to the vitreous via the solution.
[0416] The average level of rapamycin in the vitreous at 20, 40, 67, and 90 days after subconjunctival injection was about 4.425, 3.800, 4.100, and 1.500 ng/ml, respectively.
[0417] The full retina choroid was homogenized and analyzed. The average concentration of the retina choroid was calculated by dividing the mass of rapamycin measured by the mass of retina choroid analyzed. The sample did not inch.ide the site of administration; thus, this measurement indicated the level of rapa.inycin delivered to the retina choroid via the solution.

[0418] The average level of rapamycin in the retina choroid at 20, 40, 67, and 90 days after subconjunctival injection was about 0.055, 0.209, 0.080, and 0.017 ng/mg, respectively.

[04191 The sclera was analyzed in the same way as the retina choroid. The scleral sample included the site of injection; tlius, this measurement indicated clearance of rapamycin from the sclera.

[0420] The average level of rapamycin in the sclera at 20, 40, 67, and 90 days after subconjunctival injection was about 0.141, 0.271, 0.067, and 0.192 ng/mg, respectively.
Exanaple 3- Pt-epaNatioya and Characterization of a Rapanaycin-Containing Solution [0421] 5.233% rapamycin (per weight of the total of the formulation after all components were added) was dissolved in 0.4177 g of EtOH; the quantity of EtOH
was reduced by forced evaporation (heat) to 0.1296g (6.344%, w/w). PEG 400 was added under continuous agitation. Final concentrations as a percentage of the total weight were approximately: rapamycin 5.233%, ethanol 6.344%, and PEG 400 88.424%. When contacted with the vitreous, the formulation formed a non-dispersed mass relative to the surrounding medium. This solution is listed as formulation #34 in Table 1.

_--__--Exanaple 4 - Subconjunctival Injection of a Rapafnycin-Contai.ning Solution [0422] 25 l of the solution described in Example 3 were injected between the sclera and the conjunctiva of the eye of New Zealand white rabbits.

[0423] Fig. 3 depicts the level of rapamycin present in the vitreous (ng/ml), retina choroid (ng/mg), and sclera (ng/mg) on a logarithmic scale at 14, 35, 62, and 85 days after injection. The level of rapamycin present in the vitreous (ng/ml) is also shown at 2 days after injection.

[0424] The vitreous was homogenized and analyzed as described in Exainple 2, except on day 2 a single eye of each of three rabbits was analyzed; at day 14 two eyes from each of two rabbits were analyzed; at day 35 two eyes froin a single rabbit were analyzed; at day 62 two eyes from a single rabbit were analyzed; and at day 85 one eye from a single rabbit plus two eyes from a second rabbit were analyzed.

[0425] The vitreous sample did not include the site of administration; thus, this measurement indicated the level of rapamycin delivered to the vitreous via the solution.
The average level of rapamycin in the vitreous at 2, 14, 35, 62, and 85 days after subconjunctival injection was about 3.57, 53.65, 9.00, 4.700, and 0.600 ng/ml, respectively.

[0426] The retina choroid was homogenized and analyzed as described in Example 2, with the samples taken on the days as described for the vitreous above. No day 2 analysis was done. The retina choroid sample did not include the site of administration; thus, this measurement indicated the level of rapamycin delivered to the retina choroid via the soh.ition. The average level of rapamycin in the retina choroid at 14, 35, 62, and 85 days after subconjunctival injection was about 0.4815, 1.725, 0.057, and 0.009 ng/mg, respectively.

[0427] The scleral sample was analyzed as described in Example 2, and the samples were taken on the days as described for the retina choroid as above. The scleral sample included the site of injection; thus, this measurement indicated clearance of rapa.mycin from the sclera. The average level of rapamycin in the sclera at 14, 35, 62, and 85 days after subconjunctival injection was about 34.5815, 0.135, 0.042, and 0.163666667 ng/mg, respectively.

Exanaple 5 - Intravitreal Injection of a Rapanaycin-Containing Solution -------------[0428] 25 gl of the solution described in Example 3 was injected into the vitreous of the eye of New Zealand white rabbits. Fig. 4 depicts the level of rapamycin present in the vitreous (ng/ml), retina choroid (ng/mg), and sclera (ng/mg) on a logarithinic scale at 14, 35, 62, and 90 days after injection. The level of rapamycin present in the vitreous (ng/ml) is also shown at 2 days after injection.

[0429] The vitreous was homogenized and analyzed as described in Example 2, except on day 2 approximately 1 l of a single eye of each of three rabbits was analyzed; at day 14 two eyes from eacli of two rabbits were analyzed; at day 35 two eyes from a single rabbit were analyzed; at day 62 two eyes from a single rabbit were analyzed;
and at day 90 two eyes from each of two rabbits were analyzed.

[0430] Excepting the day 2 sample, the vitreous samples included the site of administration. An effort was made to avoid the administered solution where possible.
However, the accuracy of the measured levels of rapamycin was potentially affected by sampling errors due to inadvertent inclusion of the administered solution.

[0431] The average level of rapamycin in the vitreous at 2, 14, 35, 62, and 90 days after intravitreal injection was about 11.4, 136538, 2850.3, 21820.35, and 27142.75 ng/ml, respectively.

[0432] The retina choroid was homogenized and analyzed as described in Example 2, with the samples talcen on the days described for the vitreous above. No day 2 analysis was done. The retina choroid sample did not include the site of administration; thus, this measurement indicated the level of rapamycin delivered to the retina choroid via the solution. The average level of rapamycin in the retina choroid at 14, 35, 62, and 90 days after intravitreal injection was about 5.78975, 244.485, 0.105, and 1.782 ng/mg, respectively.

[0433] The scleral sample was analyzed as described in Example 2, and the samples were taken on the days as described for the retina choroid above. The scleral sample did not include the site of injection; thus, this measurement indicated level of rapamycin delivered to the sclera. The average level of rapamycin in the sclera at 14, 35, 62, and 90 days after intravitreal injection was about 0.5695, 12.34, 0.8505, and 0.71175 ng/mg, respectively.

Exanaple 6- Prepaf-ation and Chaf acterization of a Rapanaycin-Corataining Suspension [0434) 6% rapamycin (percentage of the total weight) was dispersed in_94%-PEG400_ _(percentage of the total weight). This suspension is listed as formulation #55 in Table 1.

Exattaple 7- Intravitt=eal It jection of a Rapamycin-Containing Suspension [0435] The solution prepared in Example 6 was injected intravitreally into the eyes of New Zealand white rabbits. Fig. 5 depicts images of rabbit eyes after intravitreal injection of 10 l (Fig. 5A), 20 l (Fig. 5B), and 40 l (Fig. 5C) of a 6% rapamycin suspension in PEG400. This resulted in an injected dose of about 0.6, about 1.2, and about 2.4 mg. The images were focused on the administered suspension. These images show that the suspension forms a non-dispersed mass relative to the surrounding vitreal medium.
Example 8 - Preparation and ChaYactet=ization of a Rapamycin-Containing In Situ Gelling Fornzulation [0436] A liquid formulation of 4.2% rapamycin (obtained from LC laboratories in Woburn, MA, and Chunghwa Chemical Synthesis & BioTech. Co, Ltd in Taiwan), 4.3%
ethanol (obtained from Gold Shield Chemical in Hayward, CA), 2.2% PVP K90 (obtained from BASF), 87.1 % PEG 400 (obtained from DOW Chemical), and 2.2% Eudragit RL
100 (obtained from Rohm Pharma Polymers), where all percentages are by weight of the total.

[0437] Eudragit RL 100 was dissolved in ethanol. Sonication and heat maybe required at this step. Ethanol - Eudragit was added to PEG 400. PVP was slowly added to the Eudragit-Ethanol-PEG solution, and a uniformly mixed solution was obtained.
Vigorous mixing may be required for this step.

[0438] Rapamycin was added to and dissolved in the Eudragit-ethanol-PEG-PVP
mix.
Heat and sonication may be used. The formulation was mixed thoroughly (using a vortex or mixer) to achieve uniformity. This formulation is listed as #37 in Table 1.

[0439] When placed in deionized water or tap water, the liquid formulation formed a non-dispersed mass. The non-dispersed mass appeared as a gel-like substance.

Exatnple 9 -Subconjunctival Injection of a Rapatnycin-Containing Non-Dispersed Mass-Forming Fot mulation [0440] 50 1 of the solution described in Example 8 was injected between the sclera and the conjunctiva of the eye of New Zealand white rabbits.

[0441] Fig. 6 depicts the average concentration of rapamy-cin present in_the-vitreo-us _ - _-(ng/ml), retina choroid tissues (ng/mg), and sclera (ng/mg) on a logarithmic scale at 7, 32, 45, and 90 days after injection of the in situ gelling formulation.

[0442] The analysis was by LCMS (liquid chromatography - mass spectroscopy).
[0443] Where more than a single eye was analyzed, the average concentration of rapamycin was calculated by adding the concentrations of rapamycin obtained for each eye from each rabbit, and dividing the total by the number of eyes analyzed.
In this experiment, the vitreous day 7 and the sclera day 7, 32, and 45 timepoints represent a single eye, as opposed to an average level. The remaining day 7, 32, and 45 timepoints represent the average of two eyes of one rabbit, and the day 90 timepoint represents the average of two eyes of each of two rabbits (four eyes total).

[0444] The full vitreous was homogenized and analyzed. The average concentration of the vitreous was calculated by dividing the mass of rapamycin measured by the vohime of vitreous analyzed. The sample did not include the site of administration;
thus, this measurement indicated the level of rapamycin delivered to the vitreous via the in situ gelling formulation.

[0445] The average level of rapamycin in the vitreous at 7, 32, 45, and 90 days after subconjunctival injection was about 13.9, about 7.4, about 1.35, and about 9.9 ng/ml, respectively.

[0446] The full retina choroid tissues were homogenized and analyzed. The average concentration of the retina choroid tissues was calculated by dividing the mass of rapamycin measured by the mass of retina choroid tissues analyzed. The sample did not include the site of administration; thus, this measurement indicated the level of rapamycin delivered to the retina choroid tissues via the in situ gelling formulation.

[0447] The average level of rapamycin in the retina choroid tissues at 7, 32, 45, and 90 days after subconjunctival injection was about 0.376, about 0.1875, about 0.136, and about 0.29 ng/mg, respectively.

[0448] The sclera was analyzed in the same way as the retina choroid tissues.
The scleral sample may have included the injected liquid formulation; thus, this measurement was indicative of clearance of rapamycin from the sclera.

[0449] The average level of rapamycin in the sclera at 7, 32, 45, and 90 days after subconjunctival injection was about 2033, about 1653, about 3626, and about 420.5 ng/mg, respectively.
----------- -------Example 10 - Preparation and characterization of a Rapamycin-Containing suspension.

[0450] A rapamycin containing suspension was formed by dispersing 150.5 mg of rapamycin (3.004 % by weiglit) in 4860.3 mg of PEG 400 (96.996% by weight).
This formulation is listed as #49 in Table 1. 150.5 mg rapamycin (3.004 % by weight) and 4860.3 mg of PEG 400 (96.996% by weight) were placed in an amber vial. High Wear Resistant Zirconia Grinding Media (beads) of 3 mm diameter were added, up to three quarters of the total volume. The vial was sealed and placed in a Cole-Parmer milling apparatus for 48 hrs. The particle size median for rapamycin was 2.8386 mm and the mean was 3.1275 rnrn. The formulation was kept at 4C until use. Volumes of 20 l and 40 l each formed a non-dispersed mass when placed in the vitreous of a rabbit eye.
Exanaple 11- Subconjunctival Injection of a Raparnycin-Gontaining Suspension.

[0451] 40 l of the suspension described in Example 10 were injected between the sclera and the conjunctiva of the eye of New Zealand white rabbits. Fig. 7 depicts the level of rapamycin in the vitreous (ng/ml), retina choroid (ng/mg), and the sclera (ng/mg) on a logarithmic scale at 14, 42, 63, and 91 days after injection.

[0452] The vitreous was homogenized and analyzed as described in Example 2.
Two eyes from each of two rabbits were analyzed at each time point except for day 91, on which two eyes from one rabbit were analyzed. The vitreous sample did not include the site of administration, so this measurement indicated the level of rapamycin delivered to the vitreous. The average level of rapamycin in the vitreous at 14, 42, 63, and 91 days after subconjunctival injection was about 4.031, 23.11, 53.27, and 13.94 ng/ml, respectively.

[0453] The retina choroid was homogenized and analyzed as described in Example 2, with the samples taken as described for the vitreous above. The retina choroid did not include the site of administration, so this measurement indicated the level of rapamycin delivered to the retina choroid. The average level of rapamycin in the retina choroid at 14, 42, 63, and 91 days after subconjunctival injection was about 0.1577, 4.965, 0.385, and 0.05 ng/mg, respectively.

[0454] The scleral sample was homogenized and analyzed as described in Example 2, with the samples taken as described for the vitreous above. The scleral sample included the site of injection. The average level of rapamy-cin in thesctexa. at14,-42,-63-,-and-91 days after subconjunctival injection was about 1283, 476.3, 854.2, and 168.5 ng/mg, respectively.

Exatnple 12 - Intravitreal Injection of a Rapamycin-Containing Suspension [0455] 20 l of the suspension described in Example 10 were injected into the vitreous of the eye of New Zealand white rabbits. The injected suspension formed a non-dispersed mass relative to the surrounding medium. Fig. 8 depicts the level of rapamycin in the retina choroid (ng/mg) and the sclera (ng/mg) on a logaritlunic scale at 14, 42, 63, and 91 days after injection and in the vitreous (ng/ml) at 63 and 91 days after injection.

[0456] The vitreous was homogenized and analyzed as described in Example 2.
Two eyes from each of two rabbits were analyzed at each time point. The vitreous sample may have included the site of administration. The average level of rapamycin in the vitreous at 63 and 91 days after intravitreal injection was about 381,600 and 150,400 ng/ml, respectively.
[0457] The retina choroid was homogenized and analyzed as described in Example 2.
Two eyes from each of two rabbits were analyzed at each time point. The retina choroid did not include the site of administration, so this measurement indicated the level of rapamycin delivered to the retina choroid. The average level of rapamycin in the retina choroid at 14, 42, 63, and 91 days after intravitreal injection was about 2.588, 4.249, 21.42, and 0.922 ng/mg, respectively.

[0458] The scleral sample was homogenized and analyzed as described in Example 2, with the samples talcen as described for the retina choroid above. The scleral sample did not include the site of injection, so this measurement indicated the level of rapamycin delivered to the sclera. The average level of rapamycin in the sclera at 14, 42, 63, and 91 days after intravitreal injection was about 0.7327, 6.053, 1.373, and 17.49 ng/mg, respectively.

Exafnple 13 - Preparation and Chaf-actenization of a Rapanaycin-Containing Solution.
[0459] A rapamycin containing solution was formed by placing 116.6 mg of rapainycin in ethanol and storing the mixture at 4 C for 6 hours. This solution was then mixed with 4647.5 mg of PEG 400 to give a solution having final concentrations by weight of 2.29%
rapamycin, 6.05% ethanol, and 91.66% PEG 400. This solution is listed as formulation #51-in_ Table 1-._ _ A volume_of 30 l formed. a non-dispersed-mass -when-placed -in the-------- -- ---- ---vitreous of rabbit eyes.

Example 14 - Subconjunctival Ir jection of a Rapanaycin-Containing Solution [0460] 40 l of the solution described in Example 13 were injected between the sclera and the conjunctiva of the eye of New Zealand white rabbits. Fig. 9 depicts the level of rapamycin in the vitreous (ng/ml), retina choroid (ng/mg), and the sciera (ng/mg) on a linear scale at 14, 42, 63, and 91 days after injection.

[0461] The vitreous was liomogenized and analyzed as described in Example 2.
Two eyes from each of two rabbits were analyzed at each time point except for day 91, on which two eyes from one rabbit were analyzed. The vitreous sample did not include the site of administration, so this measurement indicated the level of rapamycin delivered to the vitreous. The average level of rapamycin in the vitreous at 14, 42, 63, and 91 days after subconjunctival injection was about 1.804, 1.854, 1.785, and 1.255 ng/ml, respectively.

[0462] The retina choroid was homogenized and analyzed as described in Example 2, with the samples taken as described for the vitreous above. The retina choroid did not include the site of administration, so this measurement indicated the level of rapamycin delivered to the retina choroid. The average level of rapamycin in the retina choroid at 14, 42, 63, and 91 days after subconjunctival injection was about 1.221, 4.697, 0.1075, and 0.02 ng/mg, respectively.

[0463] The scleral sample was homogenized and analyzed as described in Example 2, with the samples taken as described for the vitreous above. The scleral sample included the site of injection. The average level of rapamycin in the sclera at 14, 42, 63, and 91 days after subconjunctival injection was about 1.987, 1.884, 0.56, and 10.84 ng/mg, respectively.

Exanzple 15 - Iyttz~avitreal IT jection of a Raparnycin-Containing Solution [0464] 30 l of the solution described in Example 13 were injected into the vitreous of the eye of New Zealand white rabbits. The injected solution formed a non-dispersed mass relative to the surrounding medium. Fig. 10 depicts the level of rapamycin in the retina choroid (ng/mg) and the sclera (ng/mg) on a linear scale at 14, 42, 63, and 91 days after injection.

[0465] The retina choroid was homogenized and analyzedasdescribed_in Example 2. -Two eyes from each of two rabbits were analyzed at each time point. The retina choroid did not include the site of administration, so this measurement indicated the level of rapamycin delivered to the retina choroid. The average level of rapamycin in the retina choroid at 14, 42, 63, and 91 days after intravitreal injection was about 5.515, 5.388, 0.3833, and 11.52 ng/mg, respectively.

[0466] The scleral sample was homogenized and analyzed as described in Exainple 2, with the samples taken as described for the retina choroid above. The scleral sample did not include the site of injection, so this measurement indicated the level of rapamycin delivered to the sclera. The average level of rapamycin in the sclera at 14, 42, 63, and 91 days after intravitreal injection was about 1.077, 0.9239, 0.0975, and 2.0825 ng/mg, respectively.

[0467] Fig. 11 depicts the level of rapamycin in the vitreous (ng/n7l) on a linear scale at 63 and 91 days after injection. The vitreous was homogenized and analyzed as described in Example 2. Two eyes from each of two rabbits were analyzed at each time point. The vitreous sample may have included the site of administration. The average level of rapamycin in the vitreous at 63 and 91 days after intravitreal injection was about 299,900 and 196,600 ng/ml, respectively.

Exa>7aple 16 - Preparation and Characterization of a Raparnycin-Containing Solution.
[0468] About 320 g of ethanol was sparged with N2 for about 10 minutes, and then about 40 g of sirolimus was added to the ethanol. The mixture was sonicated for about 20 minutes, by the end of which all of the sirolimus had gone into solution to form a sirolimus stock solution. A diluent solvent was prepared by sonicating about 1880 g of PEG 400 for about 60 minutes, and then sparging the solvent with Nitrogen for about 10 minutes.

[0469] The sirolimus stock solution and the PEG 400 were then rotated at about room temperature in a rotary evaporator for about 10 minutes to mix the stock solution with the diluent solvent. After mixing, the solution was sparged with nitrogen for about 10 minutes and blanketed with nitrogen for about 5 minutes. After the solution was sparged and filled with nitrogen, about 240 g of excess ethanol was evaporated from the solution by increasing the solution temperature, maintaining a temperature that did not exceed 40 C for an extended period of time and continuing to rotate the solution for about 2.5 hours.

[0470] The resulting solution comprised about 40 g of sirolimus (about 2% by weight), about 80 g of ethanol (about 4% by weight), and about 1880 g of PEG 400 (about 94% by weight). This solution was sparged with nitrogen for about 10 minutes and blanketed with nitrogen for about 5 minutes. The solution was then filtered through a 0.2 micron filter. HPLC vials were filled with 2 ml each of the filtered solution to leave a head space in each container of about 400 l. This head space was filled with nitrogen gas and capped.

Exainple 17 - Preparation and Chayacterization of a Rapa3nycin-Contain.ing Solution.
[0471] Rapamycin, ethanol and PEG 400 were placed in a container to give final concentrations by weight of about 2.00% rapamycin, about 4.00% ethanol, and about 94.00% PEG 400. The mixture was capped and sonicated for 1-2 hours. The sonication generated heat, with temperatures of up to about 40 or 50 C. This solution is listed as formulation #100 in Table 1. Vohunes of 1 l, 3 1, 20 1, and 40 1 formed a non-dispersed mass in the vitreous of rabbit eyes.

Exanzple 18 - Subconjunctival kjection of f a RapaMycin-Containing Solution [0472] 20 l of the solution described in Example 17 were injected between the sciera and the conjunctiva of the eye of New Zealand white rabbits. Fig. 12 depicts the level of rapamycin in the vitreous on a logarithmic scale at 5, 30, 60, 90, and 120 days after injection. Fig. 13 depicts the level of rapamycin in the retina choroid on a logarithmic scale at the same time points. For comparison, Fig. 12 and Fig. 13 also depict results of similar studies, performed with 40 1 and 60 1 injections, described below in Example 19 and Example 20.

[04731 In Figs. 12-15, discussed in this and following examples, some outlier points have been omitted. Individual data points from the same study at the same time point were compared to each other. When the arithmetic mean of the data points was lower than their standard deviation, the data points that were higher or lower by an order of magnitude were considered as outliers.

[0474] The vitreous was homogenized and analyzed as described in Example 2.
Between two and five rabbit eyes were analyzed at each time point. The vitreous sample did not include the site of adm.inistration, so this measurement indicated the level of rapamycin delivered to the vitreous. The average leYel-ofrapamycin-in the-vitr-eous at--5-, 30, 60, 90, and 120 days after subconjtinctival injection was about 1.81, 0.45, 0.39, 1.85, and 1.49 ng/ml, respectively.

[0475] The retina choroid was homogenized and analyzed as described in Example 2, with the samples taken as described for the vitreous above. The retina choroid did not include the site of administration, so this measurement iildicated the level of rapainycin delivered to the retina choroid. The average level of rapamycin in the retina choroid at 5, 30, 60, 90, and 120 days after subconjunctival injection was about 0.14, 0.03, 0.02, 0.02, and 0.01 ng/mg, respectively.

Exanaple 19 - Subcof junctival Injection of a Rapanaycin-Containing Solution [0476] 40 l of the solution described in Example 17 were injected between the sclera and the conjunctiva of the eye of New Zealand white rabbits. Fig. 12 depicts the level of rapamycin in the vitreous on a logarithmic scale at 5, 30, 60, 90, and 120 days after injection. Fig. 13 depicts the level of rapamycin in the retina choroid on a logarithmic scale at the same time points.

[04771 The vitreous was homogenized and analyzed as described in Example 2.
Between two and five rabbit eyes were a.nalyzed at each time point. The vitreous sample did not include the site of administration, so this measurement indicated the level of rapamycin delivered to the vitreous. The average level of rapamycin in the vitreous at 5, 30, 60, 90, and 120 days after subconjunctival injection was about 2.39, 0.65, 0.54, 2.07, and 1.92 ng/ml, respectively.

[0478] The retina choroid was homogenized and analyzed as described in Example 2, with the samples taken as described for the vitreous above. The retina choroid did not include the site of administration, so this measurement indicated the level of rapamycin delivered to the retina choroid. The average level of rapamycin in the retina choroid at 5, 30, 60, 90, and 120 days after subconjunctival injection was about 0.47, 0.04, 0.01, 0.05, and 0.0 ng/mg, respectively.

Exan2ple 20 - Subcor junctival Injection of a Rapaynycin-Containing Solution [0479] 60 l of the solution described in Example 17 were injected between the sclera and the conjunctiva of the eye of New Zealand white rabbits._ Fig.12 -depicts the-level of- -rapamycin in the vitreous on a logarithmic scale at 5, 30, 60, 90, and 120 days after injection. Fig. 13 depicts the level of rapamycin in the retina choroid on a logarithmic scale at the same time points.

[0480] The vitreous was homogenized and analyzed as described in Example 2.
Between two and five rabbit eyes were analyzed at each time point. The vitreous sample did not include the site of administration, so this measurement indicated the level of rapamycin delivered to the vitreous. The average level of rapamycin in the vitreous at 5, 30, 60, 90, and 120 days after subconjunctival injection was about 8.65, 0.29, 0.18, 2.00, 1.41 ng/ml, respectively.

[0481] The retina choroid was homogenized and analyzed as described in Example 2, with the samples taken as described for the vitreous above. The retina choroid did not include the site of administration, so this measurement indicated the level of rapamycin delivered to the retina choroid. The average level of rapainycin in the retina choroid at 5, 30, 60, 90, and 120 days after subconjunctival injection was about 0.63, 0.02, 0.02, 0.06, and 0.01 ng/mg, respectively.

Exafnple 21 - Intravitreal Injection of a Rapanaycin-Containing Solution [0482] 20 l of the solution described in Example 17 were injected into the vitreous of the eye of New Zealand white rabbits. The injected solution formed a non-dispersed mass relative to the surrounding medium. Fig. 14 depicts the level of rapamycin in the vitreous on a logarithmic scale 5, 30, 60, 90, and 120 days after injection. Fig. 15 depicts the level of rapamycin in the retina choroid on a logarithmic scale at the same time points. For comparison, Fig. 14 and Fig. 15 also depict results of other studies described below in Example 22 and Example 24.

[0483] The vitreous was homogenized and analyzed as described in Example 2.
Between two and five rabbit eyes were analyzed at each time point. The vitreous sample may have included the site of administration. The average level of rapamycin in the vitreous at 5, 30, 60, 90, and 120 days after intravitreal injection was about 162,100;
18,780; 57,830; 94,040; and 13,150 ng/ml, respectively.

[0484] The retina choroid was homogenized and analyzed as described in Example 2, with the samples taken as described for the vitreous above. The retina choroid did not -include-the si-te-of adininistration;-so-this measurement- indicated the-level of rapamycin delivered to the retina choroid. The average level of rapamycin in the retina choroid at 5, 30, 60, 90, and 120 days after intravitreal injection was about 2.84, 2.26, 0.17, 0.22, and 0.05 ng/mg, respectively.

Exainple 22 - Intravitreal Iiajection of a Rapainycin-Containing Solution [0485] 40 l of the solution described in Example 17 were injected into the vitreous of the eye of New Zealand white rabbits. The injected solution formed a non-dispersed mass relative to the surrounding medium. Fig. 14 depicts the level of rapanrycin in the vitreous on a logaritlunic scale 5, 30, 60, 90, and 120 days after injection. Fig. 15 depicts the level of rapamycin in the retina choroid on a logarithmic scale at the same time points.

[0486] The vitreous was homogenized aild analyzed as described in Example 2.
Between two and five rabbit eyes were analyzed at each time point. The vitreous sample may have included the site of administration. The average level of rapamycin in the vitreous at 5, 30, 60, 90, and 120 days after intravitreal injection was about 415,600;
4,830; 74,510; 301,300; and 7,854 ng/ml respectively.

[0487] The retina choroid was homogenized and analyzed as described in Example 2, with the samples taken as described for the vitreous above. The retina choroid did not include the site of administration, so this measurement indicated the level of rapamycin delivered to the retina choroid. The average level of rapainycin in the retina choroid at 5, 30, 60, 90, and 120 days after intravitreal injection was about 5.36, 0.23, 1.27, 1.08, and 0.08 ng/mg, respectively.

Exafnple 23 - Preparation and Characterization of a Rapanaycin-Containing Solution.
[0488] Rapamycin, ethanol and PEG 400 were added to a container to give final concentrations by weight of about 0.4% rapamycin, 4.0% ethanol, and 95.6% PEG
400.
The mixture was sonicated for 1-2 hours. Sonication resulted in elevated temperatures of up to about 40 to 50 C. This solution is listed as formulation #99 in Table 1.

Exanaple 24 - Intnavitreal Injection of a Rapamycin-Containing Solution [0489] 100 l of the solution described in Example 23 were injected into the vitreous of the eye of New Zealand white rabbits. The injected solution did not form a non-dispersed mass relative to the surrounding medium. Fig. 14 depicts the level ofzapamyci-n-in-the vitreous on a logarithmic scale at 5, 30, 60, 90, and 120 days after injection. Fig. 15 depicts the level of rapamycin in the retina choroid on a logarithmic scale at the saine time points.

[0490] The vitreous was homogenized and analyzed as described in Example 2.
Between two and five rabbit eyes were analyzed at each time point. The vitreous sample may have included the site of administration. The average level of rapamycin in the vitreous at 5, 30, 60, 90, and 120 days after intravitreal injection was about 151,000;
14,890; 4,743; and 1620 ng/ml respectively.

[0491] The retina choroid was homogenized and analyzed as described in Example 2, witli the samples taken as described for the vitreous above. The retina choroid did not include the site of administration, so this measurement indicated the level of rapamycin delivered to the retina choroid. The average level of rapainycin in the retina choroid at 5, 30, 60, 90, and 120 days after intravitreal injection was about 1.21, 1.84, 0.04, 0.71, and 0.0 ng/mg, respectively.

ExaYnple 25 - Preparation and CharacteYizatioiz of a Rapanzycin-Containing Solution.
[0492] A rapamycin containing solution was formed by placing 102.4 mg of rapamycin in ethanol, adding 4719.3 mg of PEG 400, and vortexing. The resulting solution had final concentrations by weight of 2.036% rapamycin, 4.154%% ethanol, and 93.81 % PEG
400.
This solution is listed as formulation #139 in Table 1.

Exafnple 26 - Subconjunctival Injection of a Rapamycin-Containing Solution [0493] 10 l of the solution described in Example 25 were injected as a single dose between the sclera and the conjunctiva of the eye of New Zealand white rabbits. Fig. 16 depicts the level of rapamycin in the vitreous on a logarithmic scale at 5 and 14 days after injection. Fig. 17 depicts the level of rapamycin in the retina choroid on a logarithmic scale at the same time points. For comparison, Fig. 16 and Fig. 17 also depict results of other studies described below in Examples 27-29.

[0494] The vitreous was homogenized and analyzed as described in Example 2.
Four rabbit eyes were analyzed at each time point. The vitreous sample did not include the site of administration, so this measurement indicated the level of rapainycin delivered to the vitreous. The average level of rapamycin in the vitreous at 5 and 14_days_after - ---- -- -subconjunctival injection was about 2.45 and 20.13 ng/ml, respectively.

[0495] The retina choroid was homogenized and analyzed as described in Example 2, with the samples taken as described for the vitreous above. The retina choroid did not include the site of administration, so this measurement indicated the level of rapamycin delivered to the retina choroid. The average level of rapamycin in the retina choroid at 5 and 14 days after subconjunctival injection was about 0.13 and 0.19 ng/mg, respectively.
Exanaple 27 - Subcof junctival It jection of a Rapanaycin-Containing Solution [0496] 60 l of the solution described in Example 25 were injected as a single dose between the sclera and the conjunctiva of the eye of New Zealand white rabbits. Fig. 16 depicts the level of rapamycin in the vitreous on a logarithmic scale at 5 and 14 days after injection. Fig. 17 depicts the level of rapamycin in the retina choroid on a logarithmic scale at the same time points.

[0497] The vitreous was homogenized and analyzed as described in Example 2.
Four rabbit eyes were analyzed at each time point. The vitreous sample did not include the site of administration, so this measurement indicated the level of rapamycin delivered to the vitreous. The average level of rapamycin in the vitreous at 5 and 14 days after subconjunctival injection was about 17.98 and 87.03 ng/ml, respectively.

[0498] The retina choroid was homogenized and aiialyzed as described in Example 2, with the samples taken as described for the vitreous above. The retina choroid did not include the site of administration, so this measurement indicated the level of rapamycin delivered to the retina choroid. The average level of rapamycin in the retina choroid at 5 and 14 days after subconjunctival injection was about 0.27 and 0.21 ng/mg, respectively.
Exarnple 28 - Subconjun.ctival Iy jection of a Rapwnycin-Containing Solution [0499] 60 l of the solution described in Example 25 were injected as two 30 l doses at two sites between the sclera and the conjunctiva of the eye of New Zealand white rabbits. Fig. 16 depicts the level of rapamycin in the vitreous on a logarithmic scale at 5 and 14 days after injection. Fig. 17 depicts the level of rapamycin in the retina choroid on a logarithmic scale at the same time points.

[0500] The vitreous was homogenized and analyzed as described in Example 2.
Four rabbit eyes were analyzed at each time point. - The v_itreous--sam.ple_did_not-i-nclude the- site-of administration, so this measurement indicated the level of rapamycin delivered to the vitreous. The average level of rapamycin in the vitreous at 5 and 14 days after subconjunctival injection was about 502.2 and 31.80 ng/ml, respectively.

[0501] The retina choroid was homogenized and analyzed as described in Example 2, with the samples taken as described for the vitreous above. The retina choroid did not include the site of administration, so this measurement indicated the level of rapamycin delivered to the retina choroid. The average level of rapamycin in the retina choroid at 5 and 14 days after subconjunctival injection was about 0.80 and 0.15 ng/mg, respectively.
Example 29 - Subconjunctival Injection of a Rapamycin-Containing Solution [0502] 90 l of the solution described in Example 25 were injected as three 30 l doses at three sites between the sclera and the conjunctiva of the eye of New Zealand white rabbits. Fig. 16 depicts the level of rapamycin in the vitreous on a logarithmic scale at 5 and 14 days after injection. Fig. 17 depicts the level of rapamycin in the retina choroid on a logarithmic scale at the same time points.

[0503] The vitreous was homogenized and analyzed as described in Example 2.
Four rabbit eyes were analyzed at each time point. The vitreous sample did not include the site of administration, so this measurement indicated the level of rapamycin delivered to the vitreous. The average level of rapamycin in the vitreous at 5 and 14 days after subconjunctival injection was about 39.05 and 13.63 ng/ml, respectively.

[0504] The retina choroid was homogenized and analyzed as described in Example 2, with the samples taken as described for the vitreous above. The retina choroid did not include the site of administration, so this measurement indicated the level of rapamycin delivered to the retina choroid. The average level of rapamycin in the retina choroid at 5 and 14 days after subconjunctival injection was about 0.83 and 0.10 ng/mg, respectively.
Example 30 - Preparation and characterization of a Rapamycin-Containing suspension.
[0505] A rapamycin containing suspension was formed by placing 201.6 mg of rapamycin (3.000 % by weight) in 6518.8 mg of PEG 400 (97.000% by weight) and vortexing. The resulting particle size was not quantified but it was large, estimated at about 10 m. This suspension is listed as formulation #147 in Table 1.

Exarnple 31 - Subconjunctival Injection of a Rapamycin-Containing Suspension [0506] 10 l of the suspension described in Example 30 were injected as a single dose between the sclera and the conjunctiva of the eye of New Zealand white rabbits. Fig. 18 depicts the level of rapamycin in the vitreous on a logarithmic scale at 5, 14, and 30 days after injection. Fig. 19 depicts the level of rapamycin in the retina choroid on a logarithmic scale at the same time points. For comparison, Fig. 18 and Fig. 19 also depict results of other studies described below in Example 32 and Example 33.

[0507] The vitreous was homogenized and analyzed as described in Example 2.
Four rabbit eyes were analyzed at each time point. The vitreous sample did not include the site of administration, so this measurement indicated the level of rapamycin delivered to the vitreous. The average level of rapamycin in the vitreous at 5, 14, and 30 days after subconjunctival injection was about 2.68, 0.90, and 5.43 ng/ml, respectively.

[0508] The retina choroid was homogenized and analyzed as described in Example 2, with the samples taken as described for the vitreous above. The retina choroid did not include the site of administration, so this measurement indicated the level of rapamycin delivered to the retina choroid. The average level of rapamycin in the retina choroid at 5, 14, and 30 days after subconjunctival injection was about 0.20, 0.06, and 1.23 ngJmg, respectively.

Exanaple 32 - Subconjunctival Ir jection of a Rapan2ycisa-Containing Suspension [0509] 30 l of the solution described in Example 30 were injected as a single dose between the sclera and the conjunctiva of the eye of New Zealand white rabbits. Fig. 18 depicts the level of rapamycin in the vitreous on a logarithmic scale at 5, 14, and 30 days after injection. Fig. 19 depicts the level of rapamycin in the retina choroid on a logarithmic scale at the same time points.

[0510] The vitreous was homogenized and analyzed as described in Example 2.
Four rabbit eyes were analyzed at each time point. The vitreous sample did not include the site of administration, so this measurernent indicated the level of rapamycin delivered to the vitreous. The average level of rapamycin in the vitreous at 5, 14, and 30 days after subconjunctival injection was about 84.55, 11.23, and 66.35 ng/ml, respectively.

[0511] The retina choroid was homogenized and analyzed as described in Example 2, with the sainples taken as described for the vitreous above. - The-retina-chor-oid-did-not include the site of administration, so this measurement indicated the level of rapamycin delivered to the retina choroid. The average level of rapamycin in the retina choroid at 5, 14, and 30 days after subconjunctival injection was about 1.09, 0.19, and 1.02 ng/mg, respectively.

Example 33 - Subcoz junctival Injection of a Rapamycin-Containing Suspension [0512] 90 l of the solution described in Example 30 were injected as three 30 l doses at three sites between the sclera and the conjunctiva of the eye of New Zealand white rabbits. Fig.18 depicts the level of rapaznycin in the vitreous on a logarithmic scale at 5, 14, and 30 days after injection. Fig. 19 depicts the level of rapamycin in the retina choroid on a logarithmic scale at the same time points.

[0513] The vitreous was homogenized and analyzed as described in Example 2.
Four rabbit eyes were analyzed at each time point. The vitreous sample did not include the site of administration, so this measurement indicated the level of rapamycin delivered to the vitreous. The average level of rapamycin in the vitreous at 5, 14, and 30 days after subconjunctival injection was about 29.95, 15.30, and 49.20 ng/ml, respectively.

[0514] The retina choroid was homogenized and analyzed as described in Example 2, with the samples tal{en as described for the vitreous above. The retina choroid did not include the site of administration, so this measurement indicated the level of rapamycin delivered to the retina choroid. The average level of rapamycin in the retina choroid at 5, 14, and 30 days after subconjunctival injection was about 0.55, 1.31, and 5.74 nglmg, respectively.

Example 34 - Preparation and Clzanacterization of a Rapamycin-Containing Solution.
[0515] 10.3 ing of rapamycin was placed in ethanol, 4995.8 mg of PEG 400 was added, and the mixture was vortexed to give a solution having final concentrations by weight of 0.205% rapamycin, 0.544% ethanol, and 99.251 % PEG 400. This solution is listed as formulation #140 in Table 1. A volume of 10 l of this solution formed a non-dispersed mass when placed in the vitreous of a rabbit eye.

Example 35 - Intravitreal Injection of a Raparnycin-Containing Solution [0516] 10 l of the solution described in ExaMDle 34were- injectedsnto_the--vitr-eous of the eye of New Zealand white rabbits. The injected solution formed a non-dispersed mass relative to the surrounding medium. Fig. 20 depicts the level of rapamycin in the retina choroid on a logarithnlic scale at 5 and 30 days after injection. Fig. 21 depicts the level of rapamycin in the vitreous on a logarithinic scale at the same timepoints. For coinparison, Fig. 20 and Fig. 21 also depict results of other studies described below in Example 37 and Example 39.

[0517] The vitreous was homogenized and analyzed as described in Exanzple 2.
Five rabbit eyes were analyzed at each time point. The vitreous sample may have included the site of administration. The average level of rapamycin in the vitreous at 5 and 30 days after intravitreal injection was about 12.02 and 0.92 ng/ml, respectively.

[05181 The retina choroid was homogenized and analyzed as described in Exanzple 2, with the samples taken as described for the vitreous above. The retina choroid did not include the site of administration, so this measurement indicated the level of rapamycin delivered to the retina choroid. The average level of rapamycin in the retina choroid at 5 and 30 days after intravitreal injection was about 0.08 and 0.02 ng/mg, respectively.
Exainple 36 - Pf eparation arad Characterizatiora of a Rapamyciia-Containing Solution.
[0519] 31.5 mg of rapamycin was placed in ethanol, 4918.9 ing of PEG 400 was added, and the solution was vortexed. Final concentrations by weight were 0.6238%
rapammycin, 1.337% ethanol, and 98.035% PEG 400. This solution is listed as formulation #142 in Table 1. The formulation was stored at 4 C until use. A volume of 10 l of this solution formed a non-dispersed mass when placed in the vitreous of a rabbit eye.

Exarnple 37 - Intravits eal.Injection of a Rapamycin-Containing Solution [0520] 10 l of the solution described in Example 36 were injected into the vitreous of the eye of New Zealand white rabbits. The injected solution formed a non-dispersed mass relative to the surrounding medium. Fig. 20 depicts the level of rapamycin in the retina choroid on a logarithmic scale at 5 and 30 days after injection. Fig. 21 depicts the level of rapamycin in the vitreous on a logarithxnic scale at the same timepoints.

[0521) The vitreous was homogenized and analyzed as described in Example 2.
Five rabbit eyes were analyzed at each time point. The vitreous sample may have included the site of administration. The average level of rapamy_cin tn the_vitreous at 5-and-30-days after intravitreal injection was about 87.46 and 44.34 ng/ml, respectively.

[0522] The retina choroid was homogenized and analyzed as described in Example 2, with the samples taken as described for the vitreous above. The retina clloroid did not include the site of administration, so this measurement indicated the level of rapamycin delivered to the retina choroid. The average level of rapamycin in the retina choroid at 5 and 30 days after intravitreal injection was about 1.40 and 0.01 ng/mg, respectively.
Example 38 - Preparation and Cltayactef=ization of a Rapaiyaycin-Containin.g Solution.
[0523] 103.5 mg of rapamycin was placed in ethanol, 4720.8 mg of PEG 400 was added, and the mixture was vortexed to give a sohition having final concentrations by weight of 2.057% rapamycin, 4.116% ethanol, and 93.827% PEG 400. This solution is listed as formulation #144 in Table 1. A volume of 10 l of this solution formed a non-dispersed mass in the vitreous of a rabbit eye.

Exafnple 39 - Intravitreal Injection of a Rapanaycin-Containin.g Solution [0524] 10 l of the solution described in Example 38 were injected into the vitreous of the eye of New Zealand white rabbits. The injected solution fomied a non-dispersed mass relative to the surrounding mediunl. Fig. 20 depicts the level of rapamycin in the retina choroid on a logarithmic scale at 5, 30, and 90 days after injection. Fig. 21 depicts the level of rapamycin in the vitreous on a logarithmic scale at the same timepoints.

[0525] The vitreous was homogenized and analyzed as described in Example 2.
Four rabbit eyes were analyzed at each time point. The vitreous sample may have included the site of administration. The average level of rapamycin in the vitreous at 5, 30, and 90 days after intravitreal injection was about 120,500; 55,160; and 0.55 ng/ml, respectively.
[0526] The retina choroid was homogenized and analyzed as described in Example 2, with the samples taken as described for the vitreous above except that five rabbit eyes were analyzed at the 5 and 30 day time points. The retina choroid did not include the site of adininistration, so this measurement indicated the level of rapamycin delivered to the retina choroid. The average level of rapamycin in the retina choroid at 5, 30, and 90 days after intravitreal injection was about 4.75, 0.17, and 0.01 ng/mg, respectively.

Exanaple 40 - Subconjunctival Injection of a Rapainycin-Cofitaiyzing S_olutiota_ _.--[0527] 40 l of the solution described in Example 17 were injected between the sclera and the conjunctiva of the eye of New Zealand white rabbits. Fig. 22 depicts on a logarithmic scale the level of rapainycin in the aqueous htunor (ng/ml) at 1, 4, 7, 11, 14, 21, 28, 35, 54, and 56 days after injection, and the levels of rapamycin in the cornea (ng/mg) and the retina choroid (ng/ing) at 4, 14, 21, and 35 days after injection. The retina choroid level is labeled as "R/Choroid" in figure 22.

[0528] The aqueous humor was homogenized and then analyzed by liquid chromatography and mass spectroscopy. Four rabbit eyes were analyzed for each time point. The aqueous humor did not include the site of injection, so this measurement indicated the level of rapamycin delivered to the aqueous humor. The average level of rapamycin in the aqueous humor at 1, 4, 7, 11, 14, 21, 28, 35, 54, and 56 days after injection was about 0.875, 1.0, 7.0, 0.725, 0.5, 0.525, 0.0, 0.125, 0.014, and 0.0485 ng/ml, respectively.

[0529] The cornea was homogenized and then analyzed by liquid chromatography and mass spectroscopy. The cornea did not include the site of injection, so this measurement indicated the level of rapamycin delivered to the cornea. Four rabbit eyes were analyzed for each time point. The average level of rapamycin in the cornea at 4, 14, 21, and 35 days after injection was about 0.3225, 0.1, 0.0275, and 0.0125 ng/mg, respectively.
[0530] The retina choroid was homogenized and analyzed as described in Example 2, with the samples taken as described for the vitreous above. The retina choroid did not include the site of administration, so this measurement indicated the level of rapamycin delivered to the retina choroid. The average level of rapamycin in the retina choroid at 4, 14, 21, and 35 days after injection was about 11.61, 0.2, 0.0275, and 2.655 ng/mg, respectively.

Exanaple 41 - Intravitreal Injection of a Rapamycin-Containing Solution [0531] 1.0 l of the solution described in Example 17 was injected into the vitreous of the eye of New Zealand white rabbits. The injected solution formed a non-dispersed mass relative to the surrounding medium. Table 2 reports the average level of rapamycin in the aqueous humor one day after injection. For comparison, Table 2 also reports results of studies described in Examples 42-45 below.

[0532] The aqueous humor was homogenized and analyzed as described in Example 40.
__Two-rabbit _ey-es-_were _analyzed.-_The-aqueous humor_ did not-incl-ude -the-site-of-i-nj ection,--so this measurement indicated the level of rapamycin delivered to the aqueous humor.

The average level of rapamycin in the aqueous humor at 1 day after injection was about 0.438 ng/ml with a standard deviation of about 0.141 ng/ml.

Example 42 - IntNavitreal Injection of a Rapamycin-Containing Solution [0533] 3.0 l of the solution described in Example 17 were injected into the vitreous of the eye of New Zealand white rabbits. The injected solution formed a non-dispersed mass relative to the surrounding medium. Table 2 reports the average level of rapamycin in the aqueous humor one day after injection.

[0534] The aqueous llumor was homogenized and analyzed as described in Example 40.
Two rabbit eyes were analyzed. The aqueous humor did not include the site of injection, so this measurement indicated the level of rapamycin delivered to the aqueous humor.
The average level of rapamycin in the aqueous humor at 1 day after injection was about 0.355 ng/ml with a standard deviation of about 0.234 mg/ml.

Example 43 - Subcof junctival Injection of a Rapanaycin-Containing Solution [0535] 3.0 gl of the solution described in Example 17 were injected between the sclera and the conjunctiva of the eye of New Zealand white rabbits. The injected solution formed a non-dispersed mass relative to the surrounding medium. Table 2 reports the average level of rapamycin in the aqueous humor one day after injection.

[0536] The aqueous humor was homogenized and analyzed as described in Example 40.
Two rabbit eyes were analyzed. The aqueous humor did not include the site of injection, so this measurement indicated the level of rapamycin delivered to the aqueous humor.
The average level of rapamycin in the aqueous humor at 1 day after injection was about 0.338 ng/ml with a standard deviation of about 0.122 ng/ml.

Exatnple 44 - AnteNior Claafnber administration of a Rapamycin-Containing Solution [0537] 5.0 l of the solution described in Example 17 were injected into the anterior chamber of the eye of New Zealand white rabbits by injection into the front-end of the eye. The aqueous humor was withdrawn using a syringe. Table 2 reports the average level of rapamycin in the aqueous humor 14 days after injection.

[0538] The aqueous humor was homogenized and-analyzed as--describedin Example--40.------------- - Two rabbit eyes were analyzed. The aqueous humor did not include the site of injection, so this measurement indicated the level of rapamycin delivered to the aqueous humor.

The average level of rapamycin in the aqueous lnimor at 14 days after injection was about 0.166 ng/ml with a standard deviation of about 0.1 83 ng/ml.

Example 45 - Anterior Claamber Adininistration of a Rapanzycin-Contairzirzg Solution [05391 10 l of the solution described in Example 17 were injected into the anterior chamber of the eye of New Zealand wliite rabbits. Table 2 reports the average level of rapamycin in the aqueous humor 14 days after injection.

[0540] The aqueous humor was homogenized and analyzed as described in Example 40.
Two rabbit eyes were analyzed. The aqueous humor did not include the site of injection, so this measurement indicated the level of rapamycin delivered to the aqueous humor.
The average level of rapamycin in the aqueous humor at 14 days after injection was about 0.004 ng/ml with a standard deviation of about 0.006 ng/ml.
[0541] All references cited herein, including patents, patent applications, and pLtblications, are hereby incorporated by reference in their entireties, whether previously specifically incorporated or not.

Table 1 Liquid Formulations Formulation Median particle Formation of NDM, Form. # Composition mg , !o (w/w Ty e size Injection volume 1 DMSO = 2000mg (20%) S
Water = 8000mg (80%) 2 F68 = 1000mg (10%) S
Water = 9000mg (90%) 3 F68 = 3000mg (30%) S
Water = 7000mg (70%) 4 F127 = 1000mg (10%) S
Water = 9000mg (90%) F127 = 1500mg (15%) S
Water = 8500mg (85%) 6 Beta-cyclodextrin = 250mg (2.5%) S
Water = 9750mg (97.5%) 7 Rapa = 10.2mg (0.101 %) S No, 50 pL
Pluronic, F68 = 1010mg (9.99%) Water = 9090mg (89.909%) 8 Rapa = 10.2mg (0.102%) S No, 50 pL
Pluronic, F68 = 3000mg (29.969%) Water = 7000mg (69.929%) 9 Rapa = 10.5mg (0.104%) S No, 50 pL
Pluronic, F127 = 1010mg (9.99%) Water = 9090mg (89.907%) Rapa = 10.5mg (0.105%) S No, 50 pL
Pluronic, F127 = 1500mg (14.984%) ._---__--_----_ __----. -Water-=-8925rri-- 84.9%0 -Formulation Median particle Formation of NDM, Form. # Composition mg ,% (w/w) Type size Injection volume 11 Rapa = 10.7mg (0.105%) S No, 50 pL
Beta-cyclodextrin = 255mg (2.497%) Water = 9945mg (97.398%) 12 Rapa = 6.4mg (0.0999%) SP
CMC = 48mg (0.7493%) Polysorbitan 20 = 2.56mg (0.04%) Water = 6349.44mg 99.111 %
13 Rapa = 6.5mg (0.0999%) S
DMSO = 325mg (4.995%) Water = 6175mg (94.905%) 14 Rapa = 13.5mg (0.0999%) SP
CMC = 101.25mg (0.7493%) Polysorbitan 20 = 5.4mg (0.04%) Water = 13393.35m (99.112%) 15 Rapa = 11.Omg (0.2%) S
EtOH = 5500mg (99.8%) 16 Rapa = 6.6mg (0.1 %) S
EtOH = 1054.6mg (15.933%) F127 = 833.64mg (12.595%) Water = 4723.96mg (71.372%) 17 Rapa = 5 mg (0.1 %) S
Cavitron = 0.25 g (5%) Ethanol, 95% = 7 mg (1.1%) Sterile water = 4.753 (93.8%) 18 Rapa = 5 mg (0.1 %) S
Ethanol, 95% =150 mg (2.9%) PEG400 = 1.0 g (19.4%) Sterile water = 4.01 (77.6%) 19 Rapa = 5 mg (0.1 %) S Yes, 50 pL
Ethanol, 95% =152 mg (3.2%) PEG400 = 1.5227 g (30.2%) Sterile water = 3.3592 (66.67%) 20 Rapa = 6.6mg (0.1 %) S
EtOH = 505.1 mg (7.618%) F127 = 917.8mg (13.843%) Water = 5200.6m (78.44%) 21 Rapa = 6.6mg (0.1 %) S No, 50 pL
EtOH = 536mg (7.5%) Pluronic, F127 = 983.75mg (14.0%) Water = 5574.56mg (78.4%) 22 Rapa = 5.2mg (0.1023%) S
EtOH = 56.6mg (1.127%) Captisol = 2008.9mg (39.5%) Water = 3013.3mg (59.3%) 23 Rapa = 6.9mg (0.201 %) S
EtOH = 3418.0m (99.799%) 24 Rapa = 9.1 mg (0.491 %) S
EtOH = 90.9mg (4.908%) F127 = 262.8mg (14.191 %) Water = 1489.1m (80.409%) 25 Rapa = 0mg (0%) S
EtOH = 310.2mg (5.144%) F127 = 858.1 mg (14.228%) Water = 4862.6m (80.628%) ---26 apa---0mg J0%)- -- _ .-- ._ _ _ - - __ _ . - --S- -- -- - -EtOH = 613.1 mg (10.19%) F127 = 810.6mg (13.471%) Water = 4593.6mg (76.339%) Formulation Median particle Formation of NDM, Form. # Composition mg ,% (w/w) Type size Injection volume 27 Rapa = 53.5mg (1.095%) S Yes, 50 pL
EtOH = 414.8mg (8.488%) F127 = 662.8mg (13.563%) Water = 3755.7mg 76.854%
28 Rapa = 0.3 g(10 ! ) ISG, SP
PVP K90 = 0.35 g (12%) Eudragit RS30D = 2.35 g (78%) 29 Rapa = 0.2154 g(7.31 %) ISG, SP
PVP K90 = 0.25 g (8.5%) Eudragit RS30D = 2.48 84.19%
30 Rapa = 53.9mg (1.103%) S No, 50 NL
EtOH = 413.6mg (8.463%) Sterile water = 3843.5mg (78.647%) F127 Lutrol = 576.0m 11.786%
31 Rapa = 0mg (0%) S
EtOH = 411.9mg (8.513%) Sterile Water = 3849.3mg (79.554%) F127 Lutrol = 577.4m 11.933%
32 Rapa = 54.1 mg (1.256%) S
EtOH = 416.8mg (9.676%) Sterile Water = 3836.3mg (78.569%) F127 Lutrol = 577.5m (10.499%) 33 Rapa = 80.7g (1.964%) S
EtOH = 65.0mg (0.158%) PEG400 = 4021.8m 97.878%
34 Rapa = 106.9g (5.233 l ) S Yes, 25 pL
EtOH = 129.6mg (6.344%) PEG400 = 1806.5m 88.424%
35 Rapa = 0 mg (0%) ISG, SP
PVP K90 = 0.204 g (2.3%) Ethanol, 100% = 0.4 g (4.5%) Eudragit RL100 =0.201g (2.3%) PEG 400 = 8.00 90.9%
36 Rapa = 0 mg (0%) ISG, SP
PVP K90 = 0.2 g (2.2%) Ethanol, 100% = 0.4 g (4.4%) PVAP = 0.4 g (4.4%) PEG400=8.00 88.9%
37 Rapa = 106.1 mg (4.2%) ISG, SP
PVP K90 = 55.2 mg (2.2%) Ethanol, 100% = 108 mg (4.3%) Eudragit RL100 = 55 mg (2.2%) PEG 400 = 2.2 87.1 %
38 Rapa = 399.6mg (9.965%) S Yes, 20 pL
F68(Lutroi) = 40.6mg (1.012%) Sterile Water = 3569.7mg (89.022%) 39 Rapa = 53.8mg (1.1 to) S
EtOH = 415.2mg (8.489%) Sterile Water = 3844.2mg (78.594%) F127 = 578.Om 11.817%
40 Rapa = 208.1 mg (3.148 to) S Yes, 20 pL
PEG400 = 6403.4m 96.852%
41 Rapa = 200.4mg (5.148 ! ) SP
F68(Lutrol) = 20.8mg (0.534%) PEG400 =3569.3rng_(91.697% -)--.- -.
EtOH 95% = 102m 2.62%

Formulation Median particle Formation of NDM, Form. # Composition mg ,% wIw Type size In"ection volume 42 Rapa = 200.4g (5.259%) SP
PEG400 = 3561.4mg (93.46%) Tween 80 = 48.8m 1.281 !
43 Rapa = 30.9 mg (1.03%) S No, 50 pL
PEG 400 = 2.9624 98.97%
44 Rapa = 61 mg (1.96%) S Yes, 50 pL
Ethanol, 100% = 0.1860 g (6%) PEG 400 = 2.8588 (92.04%) 45 Rapa = 90.7 mg (3.02%) S Yes, 50 pL
Ethanol, 100% = 0.2722 g (9.06%) PEG 400 = 2.6423 g 87.94 !
46 Rape = 101.6mg (4.997%) S
EtOH = 331.6mg (16.308%) PEG400 = 1600.1 mg (78.695%) 47 Rapa = 120.9g (3.189 !0) SP
F68(Lutrol) = 42.4mg (1.118%) Sterile Water = 3627.7m 95.692%
48 Rapa = 100.1g (1.999 l ) S
EtOH = 305.1 mg (6.092%) PEG400 = 4602.9m 91.909%
49 Rapa = 150.5mg (3.004%) SP Yes, 20 pL, 40 pL
PEG400 = 4860.3mg (96.996%) 50 Rapa = 153.4mg (3.055%) SP No, 20 pL
F68(Pluronic)= 50.6mg (1.008%) Sterile Water = 4816.6mg 95.937%
51 Rapa = 116.6mg (2.29%) S Yes, 30 pL
EtOH = 306.6mg (6.05%) PEG400 = 4647.5m 91.66%
52 Rapa = 150.4 mg (2.994%) SP
F68 Lutrol = 15.4 mg (0.306%) Sterile water = 4859.1 m 96.7%
53 Rapa = 306.5 mg (6.088%) SP
PEG 400 = 4727.7 mg 93.912%
54 Rapa = 309.3 mg (6.146%) SP
PEG 400 = 4723.3 mg 93.854%
55 Rapa = 303.3 mg (6.061 ! ) SP
PEG 400 = 4700.6 mg (93.939%) 56 Rapa = 305.4 mg (6.088%) SP
PEG 400 = 4711.0 mg 93.912%
57 Rapa = 306.9 mg (6.098%) SP
PEG 400 = 4725.5 mg (93.902%) 58 Rapa = 302.5 mg (6.021 %) SP
PEG 400 = 4721.6 mg (93.979%) 59 Rapa = 304.5 mg (6.053%) SP
PEG 400 = 4726.4 mg (93.947%) 60 Dexamethasone = 251.4 mg SP
(5.011 ! ) PEG 400 = 4765.2 mg 94.989 !
61 Dexamethasone = 252.4 mg (5%) SP
PEG 400 = 4600 mg (92%) EtOH = 150 m 3%
62 Rapa = 32.2 mg (0.641 ! ) S
PEG 400 = 4677.9 mg (93.096%) EtOH = 314.7 m 6.263%
63_ Rap.a _=_32.3 mg (0.6%) S
PEG 400 = 5516.3 mg (93.1 ! ) EtOH = 314.7 m 6.263%

Formulation Median particle Formation of NDM, Form. # Composition mg ,% w/w Type size Injection volume 64 Rapa = 54.4 mg (1.007%) S
PEG 400 = 4638.9 mg (92.702%) EtOH=314.8m 6.291%
65 Rapa = 50.8 mg (1.013%) S
PEG 400 = 4963.2 mg (98.987%) 66 Rapa = 52.1 mg (1.035%) S
PEG 400 = 4868.6 mg (96.718%) EtOH = 113.1 mg (2.247%) 67 Rapa = 50.5 mg (1.009%) S Yes, 20 pL
PEG 400 = 4752.8 mg (94.953%) No, 40 pL, 100 pL
EtOH = 202.1 mg (4.038%) 68 Rapa = 101.8 mg (2.030%) S
PEG 400 = 4712.4 mg (93.970%) EtOH = 200.6 mg (4.000%) 69 Rapa = 102.1 mg (2.036%) S
PEG 400 = 4605.5mg (91.847%) EtOH = 306.7 mg (6.117%) 70 Rapa = 101.6 mg (2.025%) S
PEG 400 = 4510.6 mg (89.892%) EtOH = 405.6 mg 8.083%
71 Rapa = 75.9 mg (3.019%) SP
PEG 400 = 2438.4 mg 96.981 %
72 Rapa = 50.9 mg (2.034%) S
PEG 400 = 2350.1 mg (93.914%) EtOH = 101.4m (4.052%) 73 Rapa = 12.5 mg (0.620%) SP
PEG 400 = 2004.8 mg (99.380%) 74 Rapa = 1.20949 g(2.0152%) S
EtOH = 2.401 g (4.000%) PEG 400 = 56.407 (93.9848%) 75 Rapa = 16.0 mg g (0.795%) S No, 50 pL
EtOH = 80.0 mg (3.976%) PEG 400 = 1916.0 mg (95.2298%) 76 Rapa = 8.1 mg (0.400%) SP
PEG 400 = 2014.5 mg (99.600%) 77 Rapa = 8.6 mg (0.428%) S
PEG 400 = 2002.5 mg (99.572%) 78 Rapa = 8.2 mg (0.410%) S
PEG 400 = 1992.0 mg (99.590%) 79 Rapa = 8.7 mg (0.433%) S
PEG 400 = 1998.8 mg (99.567%) 80 Rapa = 8.6 mg (0.427%) S
PEG 400 = 2003.2 mg (99.573%) 81 Rapa = 8.6 mg (0.428%) S
PEG 400 = 1999.3 mg (99.572%) 82 Rapa = 9.0 mg (0.448%) S
PEG 400 = 2000.8 mg (99.552%) 83 Rapa = 8.0 mg (0.397%) S
PEG 400 = 2008.8 mg (99.603%) 84 Rapa = 8.5 mg (0.422%) S
PEG 400 = 2006.8 mg (99.578%) 85 Rapa = 8.0 mg (0.399%) S
PEG 400 = 1998.2 m g 99.601 %
86 -Rapa =_8.5 mg -(0.4_22%0)--_ - -S
PEG 400 = 2004.3 mg (99.578%) 87 Rapa = 8.6 mg (0.428%) S
PEG 400 = 2002.5 mg (99.572%) Formulation Median particle Formation of NDM, Form. # Composition mg ,% w/w Type size Injection volume 88 Rapa = 0.7 g (1.983%) S
EtOH = 1.4 g (3.966%) PEG 400 = 33.2 94.051 %
89 Rapa=Og(0%) S
EtOH = 0.574 g (1.995%) PEG 400 = 28.2 (98.005%) 90 Rapa = 1.95 g (1.950%) S
EtOH = 4.05 g (4.050%) PEG 400 = 94.00 g (94000.%) 91 Rapa = 0.0107 g (0.534%) S No, 80 pL
EtOH = 0.0805 g (4.019%) PEG 400 = 1.912 (95.447%) 92 Rapa = 0.0081 g (0.403%) S No, 100 pL
EtOH = 0.0804 g (4.003%) PEG 400 = 1.920 (95.594%) 93 Rapa = 1.992 g(2%) S
EtOH = 3.9419 (4%) PEG 400 = 93.95 (94%) 94 Rapa = 0.405 g (0.4%) S
EtOH = 4.24 g (4%) PEG 400 = 95.6 (95.6%) 95 PEG 400 = 96 g (96%) S
EtOH = 3.9027 (4%) 96 Rapa = 0.4020 g (0.402%) S
EtOH = 3.970 g (3.971 %) PEG 400 = 95.600 (95.627%) 97 Rapa = 2.000 g (1.990%) S
EtOH = 4.000 g (3.980%) PEG 400 = 94.500 g (94.030%) 98 PEG 400 = 96 g(96%) S
EtOH = 3.92 (4%) 99 Rapa = 0.4036 g (0.4%) S No, 100 pL
EtOH = 3.9054 g (4%) PEG 400 = 95.6 (95.6%) 100 Rapa = 2.0025 g (2%) S Yes, 1 pL, 3 pL, 20 EtOH = 3.98 g (4%) pL, 40 pL
PEG 400 = 94.00 (94%) 101 Rapa = 9.5 mg (0.472%) S
EtOH = 90.3 mg (4.485%) PEG 600 = 1913.5 mg (95.043%) 102 Rapa = 44.6 mg (2.21 %) S
EtOH = 86.1.0 mg (4.26%) PEG 600 = 1891.1 mg (93.53%) 103 Rapa = 1.97 g(2%) S
EtOH = 4.10 g (4%) PEG 400 = 94.15 (94%) 104 Rapa = 1.95 g(2%) S
EtOH = 4.00 g (4%) PEG 400 = 94.0 94%
105 Rapa = 8.00 g (2%) S
PEG 400 = 376.0 g EtOH = 16.0 (4%) 106 Rapa = 6.00 g (2%) S
PEG 400 = 282.0 g (94%) 107 Rapa = 8.9 mg (0.4434%) S
EtOH = 80.3 mg (4.0006%) PEG 300 = 1918.0 mg (95.556%) Formulation Median particle Formation of NDM, Form. # Composition mg ,% w/w Type size Injection volume 108 Rapa = 40.8 mg (2.00886%) S
EtOH = 110.0mg (5.41605%) PEG 300 = 1880.2 mg (92.57509%) 109 Rapa = 9.9 mg (0.488%) S
EtOH = 86.7mg (4.277%) PEG 400/300(50/50) = 1930.3 mg 95.235%
110 Dexamethasone = 142.5 mg SP 0.3305 pm Yes, 30 pL
(4.994%) PEG 400 = 2710.7 mg (95.006%) 111 Dexamethasone = 134.3 mg SP >10 pm (4.891%) PEG 400 = 2611.4 m 95.109%
112 Triamcinolone = 139.2 mg (5.087%) SP 3.98 pm Yes, 30 pL
PEG 400 = 2597.4 mg 94.913%
113 Triamcinolone = 135.3 mg (5.089%) SP >10 pm PEG400=2523.5m 94.911%
114 EtOH = 206.4 mg (4.121 %) S No, 30 pL
PEG 400 = 4801.6 mg (95.879%) 115 Rapa = 43.0 mg (2.144%) SP 61.4390 pm PEG 400 = 1962.3mg 97.8567%
116 Rapa = 40.0 mg (2.001 Jo) SP 3.7128 pm PEG 400 = 1959.1 m 97.999%
117 Rapa = 42.9 mg (2.142 J ) SP 2.7313 pm PEG 400 = 1959.7mg (97.858%) 118 Rapa = 100.8 mg (2.013%) SP 4.1063 pm PEG 400 = 4906.0 mg (97.987%) 119 Rapa = 20.9 mg (0.42%) S
EtOH = 209.1 mg (4.17%) PEG 400 = 4784.9 mg 95.41 %
120 Rapa = 20.6 mg (0.41 !o) S
EtOH = 211.5mg (4.22%) Benz. Ch! = 19.1 mg (0.38%) PEG 400 = 4762.0 mg (94.99%) 121 Rapa = 20.1 mg (0.40%) S
EtOH = 211.5mg (4.22%) Benz. Ch( = 2.3 mg (0.05%) PEG 400 = 4782.3 mg (95.34%) 122 Rapa = 8.0 g(2%) S
EtOH = 16.0 g (4%) PEG 400 = 376.0 94%
123 Rapa = 351.3 mg (2.006%) S
EtOH = 2353.1 mg (4.093%) PEG 400 = 16448.2.mg 93.901 %
124 Rapa = 2.2035 g(2 Jo) S
EtOH = 4.45 g (4%) PEG 400 = 103.7 94%
125 Rapa = 515.5 mg (2.021 %) SP 18.1453Nm PEG 400 = 24,993.8 mg (97.979%) 126 Rapa = 0.3 g(2 J ) S
EtOH = 0.6 g (4%) PEG 400 = 14.1 g (94%) BHT = 0.0002 0.002%
127 Rapa=0.3g(2%) S
-tOH -0 ..- --- .6 g- 4%-- -- --- --E -PEG 400 = 14.1 g (94%) BHT = 0.00037 (0.004%) Formulation Median particle Formation of NDM, Form. # Composition mg ,% (w/w) Type size Injection volume 128 Rapa = 0.3 g(2%) S
EtOH = 0.6 g (4%) PEG 400 = 14.1 g (94%) BHT = 0.0081 (0.05%) 129 Rapa = 243.2 mg (1.869%) S
EtOH = 4.88.4 mg (3.753%) PEG 400 = 12283.3 mg (94.378%) 130 Rapa = 0.404 g (2%) S
EtOH = 0.8 g (4%) PEG 400 = 18.8 g (94%) BHT = 0.00051 (0.002%) 131 Rapa = 0.6024 g(2 /o) S
EtOH = 1.2 g (4%) PEG 400 = 28.25 (94%) 132 Rapa = 2.001 g (2%) S
EtOH = 4.05 g (4%) PEG 400 = 94.45 (94%) 133 Rapa = 0.5155 g (2.057%) S
EtOH = 1.0198 g (4.070%) PEG 400 = 23.5225 g (93.873%) 134 PEG 400 = 9.6 g (96%) S
EtOH = 0.4 (4%) 135 Rapa = 0.610 g(2%) S
EtOH = 1.2 g (4%) PEG 400 = 28.2 g (94%) 136 Rapa = 24.6 mg (1.193%) S
EtOH = 91.1 mg (4.418%) Tyloxapol = 219.6 mg (10.649%) BSS = 1726.8 mg (83.740%) 137 Rapa =100.0 mg (1.993%) SP
PEG 400 = 4916.9 mg (98.007%) 138 Rapa = 201.6 mg (4.005%) SP
PEG 400 = 4831.5 mg (95.995%) 139 Rapa = 102.4 mg (2.036%) S
EtOH = 209.0 mg (4.154%) PEG 400 = 4719.3 mg 93.810%
140 Rapa = 10.3 mg (0.205%) S Yes, 10 pL
EtOH = 27.4 mg (0.544%) PEG 400 = 4995.8 mg 99.251 %
141 Rapa = 10.6 mg (0.211 %) S No, 10 pL
EtOH = 208.4 mg (4.150%) PEG 400 = 4802.3 mg (95.639%) 142 Rapa = 31.5 mg (0.628%) S Yes, 10 pL
EtOH = 67.1 mg (1.337%) PEG 400 = 4918.9mg (98.035%) 143 Rapa = 30.8 mg (0.613%) S No, 10 pL, 100 pL
EtOH = 204.5 mg (4.073%) PEG 400 = 4786.1 mg (95.314%) 144 Rapa = 103.5 mg (2.057%) S Yes, 10 pL
EtOH = 207.1 mg (4.116%) PEG 400 = 4720.8 mg (93.827%) 145 Rapa = 283.0 mg (2.020%) S
EtOH = 566.1 mg (4.041 %) PEG 400 = 13,160.8 mg (93.939%) -_ 1-46.---- --Rapa_=280:1-mg (1:9980/6)---- S
EtOH = 565.2 mg (4.033%) PEG 400 = 13,171.7 m (93.969%) Formulation Median particle Formation of NDM, Form. # Composition m,% w/w Type size Injection volume 147 Rapa = 201.6 mg (3.000%) SP
PEG 400 = 6518.8 mg (97.000%) 148 Rapa = 31.9 mg (1.019%o) S
Benzyl Alcohol = 1021.9 mg (20.070%) Sesame Oil = 4017.9 m 78.911 %
149 Rapa = 51.5 mg (1.03%) S
Benzyl Alcohol = 259.9 mg (5.19%) Sesame Oil = 4694.3 mg (93.78%) 150 Rapa = 5.96 g (2%) S
EtOH = 12.0 g (4%) PEG 400 = 282.0 g (94%) 151 Rapa = 54.5 mg (1.07%) S
Benzyl Alcohol = 1014.3 mg (19.95%) Olive Oil = 4014.8 mg (78.98%) 152 Rapa =0 mg (0.00%) S
Benzyl Alcohol = 269.4 mg (5.421 %) Tyloxapol = 608.2 mg (12.238%) Sesame Oil = 4092.2 mg 82.341 %
153 Rapa = 76.3 mg (1.75%) S
Benzyl Alcohol = 307.0 mg (7.06%) Tyloxapol = 607.8 mg (13.97%) Sesame Oil = 3000.5 mg (68.97%) Span 80 = 63.1 mg (1.45%) EtOH = 295.5 mg (6.79%) 154 Form. # 150 = 200 g(99.998) S
BHT = 0.004 (0.002%) 155 Rapa = 51.0 mg (0.87%) S
EtOH = 642.3 mg (10.93%) Benzyl Alcohol = 431.8 mg (7.34%) Sesame Oil = 4753.7 mg (80.86%) 156 Rapa = 51.4 mg (1.03%) S
Benzyl Alcohol = 518.4 mg (10.34%) Olive Oil = 4444.7 mg (88.64%) 157 Rapa = 8.1 g (2%) S
EtOH = 16.0 g (4%) PEG 400 = 376.0 g (94%) 158 Form. # 157 = 225.00 g (99.998%) S
BHT = 0.0045 (0.002%) 159 Rapa = 8.1 g(2%) S
EtOH = 16.0 g (4%) PEG 400 = 376 94%
160 Form. # 159 = 112.0 g(99.998%) S
BHT = 0.00224 g (0.002%) 161 Form. # 159 = 112.0 g (99.998%) S
BHT = 0.0019 (0.002%) 162 Rapa = 55.4 mg (1.10%) S
EtOH = 112.7 mg (2.25%) Benzyl Alcohol = 157.8 mg (3.15%) Cotton Seed Oil = 4688.0 mg (93.50%) 163 Rapa=5.005g(1%) S
EtOH = 10.0 g (2%) PEG 400 = 485.5 g (97%) ._-164 PEG-400-=-9.82--g-(98%) S_ EtOH = 0.235 (2%) 165 Form. # 163 = 100.25 g(99.998%) S
BHT = 0.0026g (0.002%) Formulation Median particle Formation of NDM, Form. # Composition mg ,% (w/w) Type size lnjection volume 166 Rapa = 203.1 mg (2.025%) SP 2.8651 pm F68 = 30.3 mg (0.303%) Sterile Water = 9792.6 mg (97.672%) 167 Rapa = 201.4 mg (2.0005%) SP 1.0984 pm Tween 20 = 43.9 mg (0.436%) Sterile Water = 9822.8 mg (97.564%) 168 EtOH = 0.8301 g (4.144%) S
PEG 400 = 19.2014 g (95.856%) 169 Form. # 168 = 300 1 S
170 Form. # 168 = 250 l S
Form. # 154 = 50 I
171 Form. # 168 = 200 l S
Form. # 154 = 100 il 172 Form. # 168 = 150 l S
Form. # 154 = 150 I
173 Form. # 154 = 300 1 S
174 Rapa = 102.2 mg (2.041 %) SP 0.4165 pm F68 = 16.0 mg (0.32%) Sterile Water = 4889.0 mg (97.639%) 175 Rapa = 101.1 mg (2.010%) SP 0.5294 pm Tween 20 = 27.7 mg (0.551 %) Sterile Water = 4901.0 mg 97.439%
176 BSS+ = 0 l S
Sterile Water = 0 l Form. # 154 = 1000 I
177 BSS+ = 200 l SP
Sterile Water = 0 l Form. # 154 = 800 I
178 BSS+ = 400 l SP
Form. # 154 = 600 I
179 BSS+ = 500 l SP
Form. # 154 = 500 I
180 BSS+ = 600 l SP
Form. # 154 = 400 I
181 BSS+ = 800 l SP
Form. # 154 = 200 I
182 Sterile Water = 200 I SP
Form. # 154 = 800 I
183 Sterile Water = 400 l SP
Form. # 154 = 600 I
184 Sterile Water = 500 l SP
Form. # 154 = 500 I
185 Sterile Water = 600 l SP
Form. # 154 = 400 I
186 Sterile Water = 800 l SP
Form. # 154 = 200 I
187 BSS+ = 2536.9 mg (49.98%) SP 60.2075 pm Form. # 154 = 2538.7 mg (50.02%) 188 Sterile Water = 2515.6 mg (49.84%) SP 617.5157 pm Form. # 154 = 2532.2 mg (50.16%) 189 F68 = 12.6 mg (0.25%) SP 70.6089 pm Sterile Water = 2524.7 mg (49.79%) _-------------------------_-_or-m.- =--2-533.1 m- 49:96 0 Formulation Median particle Formation of NDM, Form. # Composition mg ,% w/w Type size Injection volume 190 Rapa = 2.0225 g (2%) S
EtOH = 3.65 g (4%) PEG 400 = 94.0 g (94%) BHT = 0.002 g (0.002%) 191 F68=12.1 mg SP
Sterile Water = 2558.9 mg Form. # 154 = 2556.4 mg 192 F68=19.8mg SP
Sterile Water = 2564.1 mg Form. # 154 = 25557.5 mg 193 F68 = 25.3 mg SP
Sterile Water = 2575.1 mg Form. # 154 = 2572.9 m 194 F68 = 32.4 mg SP
Sterile Water = 2572.1 mg Form. # 154 = 2562.1 mg 195 F68 = 38.3 mg SP
Sterile Water = 2563.2 mg Form. # 154 = 2573.5 m 196 F68 = 43.6 mg SP
Sterile Water = 2541.1 mg Form. # 154 = 2556.0 mg 197 F68 = 51.2 mg SP
Sterile Water = 2594.5 mg Form. # 154 = 2594.1 mg 198 PEG 400 = 1920 g(96%) S
EtOH = 80 g 4%
199 Form. # 168 = 1000 I S
200 Form. # 168 =20 0 l S
Form. # 154 = 800 1I
201 Form. # 168 = 400 l S
Form. # 154 = 600 I
202 Form. # 168 = 500 l S
Form. # 154 = 500 i 203 Form. # 168 = 600 l S
Form. # 154 = 400 I
204 Form. # 168 = 800 l S
Form. # 154 = 200 i 205 PEG 400 = 200 l S
Form. # 154 = 800 l-tl 206 PEG 400 = 400 Eil S
Form. # 154 = 600 i 207 PEG 400 = 500 l S
Form. # 154 = 500 l-LI
208 PEG400=600p1 S
Form. # 154 = 400 1I
209 PEG 400 = 800 l S
Form. # 154 = 200 I
210 Phosal 50PG = 6735.0 mg (99.002%) S
Tween 80 = 67.9 mg (0.998%) 211 Rapa = 2.0047 g (2%) S
EtOH = 4.00 g (4%) PEG 400 = 94.05 94%
--------------212 - -" PFiosaf-50PG = 20.0662 g (98.999%) S
Tween 80 = 0.2029 g 1.001 %

Formulation Median particle Formation of NDM, Form. # Composition (mg ,% (w/w) Type size Injection volume 213 Form. # 154 = 100 l S
Form. # 168 = 900 pI
214 Form. # 154 = 100 l S
Form. # 168 = 900 LI
215 Form. # 154 = 100 l S
Form. # 168 = 900 gI
216 Form. # 154 = 100 l S
PEG 400 = 900 gI
217 Form. # 154 = 100 i S
PEG 400 = 900 I
218 Form. # 154 = 100 l S
PEG 400 = 900 I
219 Form.#154=100p1 SP
BSS+ = 900 1 220 Form. # 154 = 100 l SP
BSS+ = 900 I
221 Form. # 154 = 100 l SP
BSS+ = 900 I
222 Form. # 154 = 1000 I S
223 Form. # 154 = 1000 I S
224 Form. # 154 = 100 l S
Form. # 168 = 900 I
225 Form. # 154 = 100 l S
Form. # 168 = 900 I
226 Form. # 154 = 100 l S
Form. # 168 = 900 I
227 Form. # 154 = 100 1 S
PEG 400 = 900 L) 228 Form. # 154 = 100 I S
PEG 400 = 900 I
229 Form. # 154 = 100 l S
PEG 400 = 900 I
230 Form. # 154 = 100 l SP
BSS+ = 900 I
231 Form. # 154 = 100 l SP
BSS+ = 900 I
232 Form. # 154 = 100 I SP
BSS+ = 900 1 233 Form. # 154 = 200 l S
Form. # 168 = 800 I
234 Form. # 154 = 200 l S
Form. # 168 = 800 I
235 Form. # 154 = 200 l S
Form. # 168 = 800 I
236 Form. # 154 = 200 l S
Form. # 168 = 800 I
237 Form. # 154 = 200 l S
PEG 400 = 800 I
238 Form. # 154 = 200 I S
PEG 400 = 800 I
- ----- -- ---239 Form._#__1_54__=200-- 1___--- ----SP-BSS+ = 800 I
240 Form. # 154 = 200 l SP
BSS+ = 800 I

Formulation Median particle Formation of NDM, F'orm. # Com osition mg ,% w/w Type size Injection volume 241 Form. # 154 = 200 l SP
BSS+ = 800 il 242 Form. # 154 = 100 l S No, 10 pL
Form. 168 = 900 I
243 Form. # 154 = 100 Eil S Yes, 10 pL
PEG400=900 I
244 Form. # 154 = 100 pi SP Yes, 10 pL
BSS+ = 900 I
245 Form. # 154 = 100 i SP
BSS+/CMC 0.5% = 900 i 246 Form. # 154 = 400 l S No, 10 pL
Form. # 168 = 900 I
247 Form. # 154 = 400 l S Yes, 10 pL
PEG 400 = 900 1 248 Form. # 154 = 400 pi SP Yes, 10 NL
BSS+ = 900 1 249 Form. # 154 = 400 l SP
BSS+/CMC 0.5% = 900 ti 250 Form. # 154 = 100 l SP
BSS+/CMC 0.5% = 900 I
251 Form. # 154 = 100 l SP
BSS+/CMC 0.5% = 900 f 252 Form. # 154 =100 l SP
BSS+/CMC 0.5% = 900 11I
253 Form. # 154 = 200 ! SP
BSS+/CMC 0.5% = 800 I
254 Form. # 154 = 200 I SP
BSS+/CMC 0.5% = 800 I
255 Form. # 154 = 200 l SP
BSS+/CMC 0.5 !Q = 800 I
256 Form. # 154 = 400 l SP
BSS+/CMC 0.5 !0 = 900 i 257 Form. # 154 = 400 ~Ll SP
BSS+/CMC 0.5 /a = 900 I
258 Form. # 154 = 400 ! SP
BSS+/CMC 0.5% = 900 I
259 EtOH = 17.1 mg (0.57%) S
PEG 400 = 2997.3 m 99.43%
260 EtOH = 40.8 mg (1.35%) S
PEG 400 = 2980.2 m (98.65%) 261 EtOH = 47.1 mg (1.57%) S
PEG 400 = 2950.1 m (98.43%) 262 Rapa = 2.0032 g(2 !0) S
EtOH = 3.92 g (4%) PEG 400 = 94.00 (94%) 263 Triamcinolone acetomide = 80.8 mg SP
(4.04%) PEG 400 = 1920.8 m 95.96%
264 NFF-0007 filled in glove box S
265 PEG 400 = 9.598 g(96%) S
EtOH = 0.4052 4%
266 Triamcinolone acetomide - 42.2 m _____-_ -(4.1-23%) PEG 400 = 981.3 m 95.877%

Formulation Median particle Formation of NDM, Form. # Composition mg ,% (w/w) Type size Injection volume 267 Phosal 50PG = 20.0783 g S
(99.00835%) Tween 80 = 0.2011 (0.99165%) 268 PEG 400 = 96.1 g (96%) S
EtOH = 4.00 (4%) 269 Rapa = 0.4001 g (2%) S
EtOH = 0.80 g (4%) PEG 400 = 18.8 (94%) 270 Sterile Water = 9955.8 mg (99.27%) S
CMC High visc. = 47.8 mg (0.48%) Tween 80 = 25.4 mg (0.25%) 271 Sterile Water = 9947.5 mg (99.00%) S
CMC Medium visc. = 75 mg (0.75%) Tween 80 = 25.1 mg (0.25%) 272 Rapa = 41 mg (2.01%) SP
Form. # 270 = 2000 mg (97.99%) 273 Rapa = 40.2 mg (1.97%) SP
MSF-03-172-07E = 2000 mg (98.03%) 274 NMP (Pharmasolve = 1280.5 mg S
(65.89%) PLGA 75/25 = 662.9 mg 34.11 %
275 NMP (Pharmasolvefl = 1573.3 mg S
(80.50%) PLGA 75/25 = 381.0 mg (19.50%) 276 NMP (PharmasolveR = 1009.7 mg S Yes, 10 pL
49.8%) PLGA 75/25 = 1001.6 mg (50.20%) 277 Sterile Water = 14934.0 mg (99.25%) S
CMC Medium visc. = 112.4 mg (0.75%) 278 Propylene Glycol = 1893.7 mg S Yes, 10 pL
(93.85%) EtOH = 83.8 mg (4.16%) Ra a= 40.2 mg (1.99%) 279 Propylene Glycol = 1946.2 mg S Yes, 10 pL
(95.68%) Benzyl Alcohol = 47.1 mg (2.31 %) Ra a=40.8m 2.01%
280 PEG 300 = 1894.1 mg (93.74%) S Yes, 10 pL
EtOH = 40.1 mg (1.98%) Ra a= 86.4 mg (4.28%) 281 PEG 300 = 1925.5 mg (95.88%) S Yes, 10 pL, 30 pL
EtOH = 39.8 mg (1.98%) Ra a= 43.0 m (2.14%) 282 Rapa = 100.6 mg (2.01 %) SP Yes, 10 pL, 30 pL
MSF-03-176-02 = 4910.8 mg (97.99%) 283 Rapa = 11.5 mg (0.57%) S
PEG 300 = 2012.5 mg (99.43%) 284 Rapa = 10.3 mg (0.51 %) S
PEG 400 = 2017.2 m (99.49%) 285 Rapa = 9.8 mg (0.486%) S
PEG 600 = 2005.9 mg 99.51 %
286 Tacrolimus = 42.7 mg (2.11%) S
---EtOH = 46:0 mg_ .2.27% - -PG = 1938.7 mg (95.62%) Formulation Median particle Formation of NDM, Form. # Composition mg ,% (w/w) Type size Injection volume 287 Tacrolimus = 40.7 mg (2.01 %) S
EtOH = 43.0 mg (2.12%) PEG 300 = 1942.1 mg (95.87%) 288 Tacrolimus = 40.3 mg (1.99%) S
EtOH = 43.8 mg (2.16%) PEG 400 = 1942.3 mg (95.85%) 289 Tacrolimus = 40.8 mg (2.03%) S
EtOH = 44.5 mg (2.21 %) PEG 600 = 1924.0m (95.76%) 290 Rapa = 61.0 mg (3.17 l0) S
NMP = 1226.54 mg (63.80%) PLGA 75/25 = 634.96 mg (33.03%) 291 Rapa = 100.2 mg (5.13%) S
NMP = 1492.95 mg mg (76.37%) PLGA 75/25 = 361.65 mg (18.50%) 292 Rapa = 62.9 mg (3.04%) S
NMP = 1103.8g mg (53.40%) PLGA 75/25 = 900.2 mg (43.56%) 293 Rapa = 62.4 mg (3.00%) S
NMP = 1205.1 mg mg (58.11 %) PLGA 75/25 = 806.4 mg (38.89%) 294 Sterile Water+1 % CMC Med. = 4909.1 SP
mg (97.99%) Ra mg 2.01%
295 Sterile Water+1 % CMC high. = 4903.8 SP
mg (97.96%) Ra a=101.9m (2.04%) 296 Rapa = 40.5 mg (2.03%) S
NMP = 1958.7 mg (97.97%) 297 Rapa = 20.5mg (2.0%) SP
DMA = 41.4mg (4.0%) PVP = 35.0mg (3.4%) H20 = 934.7mg (90.6%) 298 Rapa = 10.6mg (2.0%) S
DMA = 10.6mg (2.0%) PEG 400 = 506.1 m (96%) 299 Rapa = 5.2mg (2.0%) SP
1% DMA in PEG 400 = 257.4 mg (98%) 300 Rapa = 20.0mg (2.0%) S
DMA = 7.8mg (0.8%) PEG 400 = 974mg (97.2%) 301 Rapa = 20.1mg (1.3%) S
DMA = 19.5mg (1.3%) PEG 400 = 1449.6mg (97.3%) 302 Rapa = 20.0mg (2.0%) SP
PVP = 10.8mg (1.1 %) PEG 400 = 994.5mg (97.0%) 303 Rapa = 20.4mg (2.0%) SP
PVP = 24.5mg (2.4%) PEG 400 = 990.7mg (95.7%) 304 Rapa = 25.5mg (2.4%) SP
PVP = 51.9mg (4.8%) PEG 400 = 1000.6mg (92.8%) -----305- --Rapa-22~5mg- (2.-3%)--BA = 27.5mg (2.7%) PEG 400 = 950.7mg (95.0%) Formulation Median particle Formation of NDM, Form. # Com osition mg ,% w/w T e size Injection volume 306 Rapa = 30.2mg (2.3%) SP
PVP = 240.9mg (18.6%) PEG 400 = 1021.2m (79.0%) 307 Rapa = 8.7mg (3.1 %) SP
1oloPVPin H20=273m 96.9%
308 Rape = 12.6mg (2.53%) SP
5% PVP in H20 = 501.6 mg (97.5%) 309 Rapa = 20.3mg (3.8%) SP

10% PVP in H20 = 513.9 mg (96.2%) 310 Rape = 100.5mg (2.0%) S Yes, 10 pL
DMA = 67.8mg (1.4%) PEG 400 = 4838.3mg (96.6%) 311 Rapa = 96.8mg (1.9%) S Yes, 10 pL
BA = 157.5mg (3.2%) PEG 400 = 4748.7mg (94.9%) 312 Rapa = 105.8mg (2.10/0) S
DMA = 5.63mg (0. 1%) PEG 400 = 4888.9m (97.8%) 313 Rapa = 20.2mg (2.0 /0) SP
PVP = 99.2mg (9.9%) H2O = 882.3m 88.1 lo 314 Rapa = 100.3mg (2.0%) SP
PVP = 251.4mg (5.0%) H20 = 4662.8m 93.0 l0 315 Rapa = 20.3mg (2.0 /0) S
DMA = 983.9mg (98%) 316 Triamcinolone = 22.8mg (2.0%) S Yes, 10 pL
DMA= 12.Omg (1.1 l0) PEG 400 = 1104.5mg 96.9%
317 Triamcinolone = 1.0mg (0.1%) S
EtOH = 49.30mg (4.0%) PEG 400 = 1191.9m 96.0%
318 Triamcinolone = 18.7mg (0.9%) S
PEG 400 = 959.8m 99.1%
319 Triamcinolone = 25.5mg (1.3%) S
EtOH = 83.0mg (4.1 %) PEG 400 = 1905.6m (94.6%) 320 Dexamethasone = 20.4mg (1.2%) S
EtOH = 71.7mg (4.1 %) PEG 400 = 1737.6m (98.8%) 321 Dexamethasone = 27.5mg (2.0%) S Yes, 10 pL
DMA = 5.6mg (0.4%) PEG 400 = 1347.3mg (97.6%) 322 Rapa = 9.1 mg (0.152 l0) E
EtOH = 90.9mg (1.514%) F127 = 262.8mg (4.378%) Water = 1489.1 mg (24.804%) Sesame oil = 4151.5m 69.152%
323 Rapa = 24.4mg (0.625%) E
Phosal 5OPG = 203.1 mg (5.2010l0) EtOH = 166.8mg (4.272%) Labrafac CC = 1502.8mg (38.486%) Sesame oil = 2007.7mg 51.416%
324 Form. # 174 with 2mm beads SP 0.4929 m -----325 Form. # 175with 2mm--beads --SP --- ---0:4804- m Table 2 Aqueous Humor Rapa Concentration Injection of 2% Rapa-PEG-EtOH Mean Rapa concentration (ng/mL) Standard deviation Solution (ng/nL) 1.0 L intravitreal 0.438 (1 day after injection) 0.141 3.0 L intravitreal 0.355 (1 day after injection) 0.234 3.0 L sub-conj 0.338 (1 day after injection) 0.122 5.0 L into anterior chamber 0.167 (14 days after injection) 0.183 10.0 L into anterior chamber 0.004 (14 days after injection) 0.006

Claims (27)

1. A liquid formulation comprising a therapeutic agent, wherein the liquid formulation when injected into the vitreous of a rabbit eye delivers an amount of the therapeutic agent sufficient to achieve, for a period of time of at least 30 days following administration of the liquid formulation, an average concentration of therapeutic agent in the retina choroid tissues of the rabbit eye equivalent to a rapamycin concentration of at least 0.01 ng/mg.

2. A liquid formulation comprising a therapeutic agent, wherein the liquid formulation when injected into the vitreous of a rabbit eye delivers an amount of the therapeutic agent sufficient to achieve, for a period of time of at least 30 days following administration of the liquid formulation, an average concentration of therapeutic agent in the vitreous of the rabbit eye equivalent to a rapamycin concentration of at least 1000 ng/ml.

3. A liquid formulation comprising a therapeutic agent, wherein the liquid formulation when injected between the sclera and conjunctiva of a rabbit eye delivers an amount of the therapeutic agent sufficient to achieve, for a period of time of at least 30 days following administration of the liquid formulation, an average concentration of therapeutic agent in the vitreous of the rabbit eye equivalent to a rapamycin concentration of at least 0.01 ng/ml.

4. A liquid formulation comprising a therapeutic agent,, wherein the liquid formulation when injected between the sclera and conjunctiva of a rabbit eye delivers an amount of the therapeutic agent sufficient to achieve, for a period of time of at least 30 days following administration of the liquid formulation, an average concentration of therapeutic agent in the retina choroid tissues of the rabbit eye equivalent to a rapamycin concentration of at least 0.001 ng/mg.

5. A liquid formulation comprising a therapeutic agent, wherein the liquid formulation forms a non-dispersed mass when injected into the vitreous of a rabbit eye.

6. The liquid formulation of claim 1, wherein the therapeutic agent is selected from the group consisting of rapamycin, SDZ-RAD, tacrolimus, everolimus, pimecrolimus, CCI-779, AP23841, ABT-578, cyclophilins, FK506-binding proteins (FKBPs), TAFA-93, RAD-001, temsirolimus, AP23573, 7-epi-rapamycin, 7-thiomethyl-rapamycin, 7-epi-trimethoxyphenyl-rapamycin, 7-epi-thiomethyl-rapamycin, 7-demethoxy-rapamycin, 32-demethoxy-rapamycin, 2-desmethyl-rapamycin, monoester derivatives of rapamycin, diester derivatives of rapamycin, 27-oximes of rapamycin; 42-oxo analogs of rapamycin; bicyclic rapamycins; rapamycin dimers; silyl ethers of rapamycin; rapamycin arylsulfonates, rapamycin sulfamates, monoesters at positions 31 and 42, diesters at positions 31 and 42, 30-demethoxy rapamycin, and pharmaceutically acceptable prodrugs, analogs, salts and esters thereof.

7. The liquid formulation of claim 6, wherein the therapeutic agent is selected from the group consisting of rapamycin, SDZ-RAD, tacrolimus, everolimus, pimecrolimus, CCI-779, AP23841, ABT-578, and pharmaceutically acceptable salts and esters thereof.

8. The liquid formulation of claim 2, wherein the therapeutic agent is selected from the group consisting of rapamycin, SDZ-RAD, tacrolimus, everolimus, pimecrolimus, CCI-779, AP23841, ABT-578, cyclophilins, FK506-binding proteins (FKBPs), TAFA-93, RAD-001, temsirolimus, AP23573, 7-epi-rapamycin, 7-thiomethyl-rapamycin, 7-epi-trimethoxyphenyl-rapamycin, 7-epi-thiomethyl-rapamycin, 7-demethoxy-rapamycin, 32-demethoxy-rapamycin, 2-desmethyl-rapamycin, monoester derivatives of rapamycin, diester derivatives of rapamycin, 27-oximes of rapamycin; 42-oxo analogs of rapamycin; bicyclic rapamycins; rapamycin dimers; silyl ethers of rapamycin; rapamycin arylsulfonates, rapamycin sulfamates, monoesters at positions 31 and 42, diesters at positions 31 and 42, 30-demethoxy rapamycin, and pharmaceutically acceptable prodrugs, analogs, salts and esters thereof.

9. The liquid formulation of claim 8, wherein the therapeutic agent is selected from the group consisting of rapamycin, SDZ-RAD, tacrolimus, everolimus, pimecrolimus, CCI-779, AP23841, ABT-578, and pharmaceutically acceptable salts and esters thereof.

10. The liquid formulation of claim 3, wherein the therapeutic agent is selected from the group consisting of rapamycin, SDZ-RAD, tacrolimus, everolimus, pimecrolimus, CCI-779, AP23841, ABT-578, cyclophilins, FK506-binding proteins (FKBPs), TAFA-93, RAD-001, temsirolimus, AP23573, 7-epi-rapamycin, 7-thiomethyl-rapamycin, 7-epi-trimethoxyphenyl-rapamycin, 7-epi-thiomethyl-rapamycin, 7-demethoxy-rapamycin, 32-demethoxy-rapamycin, 2-desmethyl-rapamycin, monoester derivatives of rapamycin, diester derivatives of rapamycin, 27-oximes of rapamycin; 42-oxo analogs of rapamycin; bicyclic rapamycins; rapamycin dimers; silyl ethers of rapamycin; rapamycin arylsulfonates, rapamycin sulfamates, monoesters at positions 31 and 42, diesters at positions 31 and 42, 30-demethoxy rapamycin, and pharmaceutically acceptable prodrugs, analogs, salts and esters thereof.

11. The liquid formulation of claim 10, wherein the therapeutic agent is selected from the group consisting of rapamycin, SDZ-RAD, tacrolimus, everolimus, pimecrolimus, CCI-779, AP23841, ABT-578, and pharmaceutically acceptable salts and esters thereof.

12. The liquid formulation of claim 4, wherein the therapeutic agent is selected from the group consisting of rapamycin, SDZ-RAD, tacrolimus, everolimus, pimecrolimus, CCI-779, AP23841, ABT-578, cyclophilins, FK506-binding proteins (FKBPs), TAFA-93, RAD-001, temsirolimus, AP23573, 7-epi-rapamycin, 7-thiomethyl-rapamycin, 7-epi-trimethoxyphenyl-rapamycin, 7-epi-thiomethyl-rapamycin, 7-demethoxy-rapamycin, 32-demethoxy-rapamycin, 2-desmethyl-rapamycin, monoester derivatives of rapamycin, diester derivatives of rapamycin, 27-oximes of rapamycin; 42-oxo analogs of rapamycin; bicyclic rapamycins; rapamycin dimers; silyl ethers of rapamycin; rapamycin arylsulfonates, rapamycin sulfamates, monoesters at positions 31 and 42, diesters at positions 31 and 42, 30-demethoxy rapamycin, and pharmaceutically acceptable prodrugs, analogs, salts and esters thereof.

13. The liquid formulation of claim 12, wherein the therapeutic agent is selected from the group consisting of rapamycin, SDZ-RAD, tacrolimus, everolimus, pimecrolimus, CCI-779, AP23841, ABT-578, and pharmaceutically acceptable salts and esters thereof.

14. The liquid formulation of claim 5, wherein the therapeutic agent is selected from the group consisting of rapamycin, SDZ-RAD, tacrolimus, everolimus, pimecrolimus, CCI-779, AP23841, ABT-578, cyclophilins, FK506-binding proteins (FKBPs), TAFA-93, RAD-001, temsirolimus, AP23573, 7-epi-rapamycin, 7-thiomethyl-rapamycin, 7-epi-trimethoxyphenyl-rapamycin, 7-epi-thiomethyl-rapamycin, 7-demethoxy-rapamycin, 32-demethoxy-rapamycin, 2-desmethyl-rapamycin, monoester derivatives of rapamycin, diester derivatives of rapamycin, 27-oximes of rapamycin; 42-oxo analogs of rapamycin; bicyclic rapamycins; rapamycin dimers; silyl ethers of rapamycin; rapamycin arylsulfonates, rapamycin sulfamates, monoesters at positions 31 and 42, diesters at positions 31 and 42, 30-demethoxy rapamycin, and pharmaceutically acceptable prodrugs, analogs, salts and esters thereof.

15. The liquid formulation of claim 14, wherein the therapeutic agent is selected from the group consisting of rapamycin, SDZ-RAD, tacrolimus, everolimus, pimecrolimus, CCI-779, AP23841, ABT-578, and pharmaceutically acceptable salts and esters thereof.

16. A method for treating wet age-related macular degeneration in a human subject, the method comprising administering to the human subject by intraocular or periocular delivery a volume of the liquid formulation of any of claims 7, 9, 11, 13, and 15 containing an amount of rapamycin effective to treat wet age-related macular degeneration in the human subject.

17. A method for preventing wet age-related macular degeneration in a human subject, the method comprising administering to the human subject by intraocular or periocular delivery a volume of the liquid formulation of any of claims 7, 9, 11, 13, and 15 containing an amount of rapamycin effective to prevent wet age-related macular degeneration in the human subject.

18. The method of claim 16, wherein the liquid formulation further comprises polyethylene glycol and wherein the volume of liquid formulation is administered to the human subject by placement in the vitreous and the volume of liquid formulation contains less than 100 µL of polyethylene glycol.

19. The method of claim 16, wherein the liquid formulation further comprises polyethylene glycol and wherein the volume of liquid formulation is administered to the human subject by placement between the sclera and conjunctiva and the volume of liquid formulation contains less than 150 µL of polyethylene glycol.

20. The method of claim 17, wherein the liquid formulation further comprises polyethylene glycol and wherein the volume of liquid formulation is administered to the human subject by placement between the sclera and conjunctiva and the volume of liquid formulation contains less than 150 µL of polyethylene glycol.

21. A method for treating wet age-related macular degeneration in a human subject, the method comprising administering to the human subject by intraocular or periocular delivery of a liquid formulation comprising an effective amount of a therapeutic agent selected from rapamycin, SDZ-RAD, tacrolimus, everolimus, pimecrolimus, CCI-779, AP23841, ABT-578, and pharmaceutically acceptable salts and esters thereof; and wherein the liquid formulation has one or more characteristics selected from the group consisting of (1) the liquid formulation when injected into the vitreous of a rabbit eye delivers an amount of the therapeutic agent sufficient to achieve, for a period of time of at least 30 days following administration of the liquid formulation, an average concentration of therapeutic agent in the retina choroid tissues of the rabbit eye equivalent to a rapamycin concentration of at least 0.01 ng/mg; (2) the liquid formulation when injected into the vitreous of a rabbit eye delivers an amount of the therapeutic agent sufficient to achieve, for a period of time of at least 30 days following administration of the liquid formulation, an average concentration of therapeutic agent in the vitreous of the rabbit eye equivalent to a rapamycin concentration of at least 1000 ng/ml; (3) the liquid formulation when injected between the sclera and conjunctiva of a rabbit eye delivers an amount of the therapeutic agent sufficient to achieve, for a period of time of at least 30 days following administration of the liquid formulation, an average concentration of therapeutic agent in the vitreous of the rabbit eye equivalent to a rapamycin concentration of at least 0.01 ng/ml; (4) the liquid formulation when injected between the sclera and conjunctiva of a rabbit eye delivers an amount of the therapeutic agent sufficient to achieve, for a period of time of at least 30 days following administration of the liquid formulation, an average concentration of therapeutic agent in the retina choroid tissues of the rabbit eye equivalent to a rapamycin concentration of at least 0.001 ng/mg; and (5) the liquid formulation forms a non-dispersed mass when injected into the vitreous of a rabbit eye.

22. A method for preventing wet age-related macular degeneration in a human subject, the method comprising administering to the human subject by intraocular or periocular delivery of a liquid formulation comprising an effective amount of a therapeutic agent selected from rapamycin, SDZ-RAD, tacrolimus, everolimus, pimecrolimus, CCI-779, AP23841, ABT-578, and pharmaceutically acceptable salts and esters thereof; and wherein the liquid formulation has one or more characteristic selected from the group consisting of (1) the liquid formulation when injected into the vitreous of a rabbit eye delivers an amount of the therapeutic agent sufficient to achieve, for a period of time of at least 30 days following administration of the liquid formulation, an average concentration of therapeutic agent in the retina choroid tissues of the rabbit eye equivalent to a rapamycin concentration of at least 0.01 ng/mg; (2) the liquid formulation when injected into the vitreous of a rabbit eye delivers an amount of the therapeutic agent sufficient to achieve, for a period of time of at least 30 days following administration of the liquid formulation, an average concentration of therapeutic agent in the vitreous of the rabbit eye equivalent to a rapamycin concentration of at least 1000 ng/ml; (3) the liquid formulation when injected between the sclera and conjunctiva of a rabbit eye delivers an amount of the therapeutic agent sufficient to achieve, for a period of time of at least 30 days following administration of the liquid formulation, an average concentration of therapeutic agent in the vitreous of the rabbit eye equivalent to a rapamycin concentration of at least 0.01 ng/ml; (4) the liquid formulation when injected between the sclera and conjunctiva of a rabbit eye delivers an amount of the therapeutic agent sufficient to achieve, for a period of time of at least 30 days following administration of the liquid formulation, an average concentration of therapeutic agent in the retina choroid tissues of the rabbit eye equivalent to a rapamycin concentration of at least 0.001 ng/mg; and (5) the liquid formulation forms a non-dispersed mass when injected into the vitreous of a rabbit eye.

23. The method of claim 22, wherein the human subject is identified as being at heightened risk of developing wet age-related macular degeneration in the eye to which the liquid formulation is administered.

24. The method of claim 23, wherein the human subject has dry age-related macular degeneration in at least one eye.

25. The method of claim 23, wherein the human subject has wet age-related macular degeneration in one eye and the liquid formulation is administered to the eye without wet age-related macular degeneration.

26. A method for treating dry age-related macular degeneration in a human subject, the method comprising administering to the human subject by intraocular or periocular delivery of a liquid formulation comprising an effective amount of a therapeutic agent selected from rapamycin, SDZ-RAD, tacrolimus, everolimus, pimecrolimus, CCI-779, AP23841, ABT-578, and pharmaceutically acceptable salts and esters thereof; and wherein the liquid formulation has one or more characteristic selected from the group consisting of (1) the liquid formulation when injected into the vitreous of a rabbit eye delivers an amount of the therapeutic agent sufficient to achieve, for a period of time of at least 30 days following administration of the liquid formulation, an average concentration of therapeutic agent in the retina choroid tissues of the rabbit eye equivalent to a rapamycin concentration of at least 0.01 ng/mg; (2) the liquid formulation when injected into the vitreous of a rabbit eye delivers an amount of the therapeutic agent sufficient to achieve, for a period of time of at least 30 days following administration of the liquid formulation, an average concentration of therapeutic agent in the vitreous of the rabbit eye equivalent to a rapamycin concentration of at least 1000 ng/ml; (3) the liquid formulation when injected between the sclera and conjunctiva of a rabbit eye delivers an amount of the therapeutic agent sufficient to achieve, for a period of time of at least 30 days following administration of the liquid formulation, an average concentration of therapeutic agent in the vitreous of the rabbit eye equivalent to a rapamycin concentration of at least 0.01 ng/ml; (4) the liquid formulation when injected between the sclera and conjunctiva of a rabbit eye delivers an amount of the therapeutic agent sufficient to achieve, for a period of time of at least 30 days following administration of the liquid formulation, an average concentration of therapeutic agent in the retina choroid tissues of the rabbit eye equivalent to a rapamycin concentration of at least 0.001 ng/mg; and (5) the liquid formulation forms a non-dispersed mass when injected into the vitreous of a rabbit eye.

27. A method for preventing wet age-related macular degeneration in a human subject having dry age-related macular degeneration, the method comprising administering to a human subject having dry age-related macular degeneration a liquid formulation comprising an effective amount of a therapeutic agent selected from rapamycin, SDZ-RAD, tacrolimus, everolimus, pimecrolimus, CCI-779, AP23841, ABT-578, and pharmaceutically acceptable salts and esters thereof;
wherein the liquid formulation is administered by intraocular or periocular delivery; and wherein the liquid formulation has one or more characteristic selected from the group consisting of (1) the liquid formulation when injected into the vitreous of a rabbit eye delivers an amount of the therapeutic agent sufficient to achieve, for a period of time of at least 30 days following administration of the liquid formulation, an average concentration of therapeutic agent in the retina choroid tissues of the rabbit eye equivalent to a rapamycin concentration of at least 0.01 ng/mg; (2) the liquid formulation when injected into the vitreous of a rabbit eye delivers an amount of the therapeutic agent sufficient to achieve, for a period of time of at least 30 days following administration of the liquid formulation, an average concentration of therapeutic agent in the vitreous of the rabbit eye equivalent to a rapamycin concentration of at least 1000 ng/ml;
(3) the liquid formulation when injected between the sclera and conjunctiva of a rabbit eye delivers an amount of the therapeutic agent sufficient to achieve, for a period of time of at least 30 days following administration of the liquid formulation, an average concentration of therapeutic agent in the vitreous of the rabbit eye equivalent to a rapamycin concentration of at least 0.01 ng/ml; (4) the liquid formulation when injected between the sclera and conjunctiva of a rabbit eye delivers an amount of the therapeutic agent sufficient to achieve, for a period of time of at least 30 days following administration of the liquid formulation, an average concentration of therapeutic agent in the retina choroid tissues of the rabbit eye equivalent to a rapamycin concentration of at least 0.001 ng/mg;
and (5) the liquid formulation forms a non-dispersed mass when injected into the vitreous of a rabbit eye.