WO1998054308A2 - Polyketides and their synthesis and use - Google Patents
Polyketides and their synthesis and use Download PDFInfo
- Publication number
- WO1998054308A2 WO1998054308A2 PCT/GB1998/001559 GB9801559W WO9854308A2 WO 1998054308 A2 WO1998054308 A2 WO 1998054308A2 GB 9801559 W GB9801559 W GB 9801559W WO 9854308 A2 WO9854308 A2 WO 9854308A2
- Authority
- WO
- WIPO (PCT)
- Prior art keywords
- rapamycin
- gene
- gene cluster
- precursor
- cluster
- Prior art date
Links
Classifications
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07D—HETEROCYCLIC COMPOUNDS
- C07D498/00—Heterocyclic compounds containing in the condensed system at least one hetero ring having nitrogen and oxygen atoms as the only ring hetero atoms
- C07D498/12—Heterocyclic compounds containing in the condensed system at least one hetero ring having nitrogen and oxygen atoms as the only ring hetero atoms in which the condensed system contains three hetero rings
- C07D498/18—Bridged systems
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P43/00—Drugs for specific purposes, not provided for in groups A61P1/00-A61P41/00
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07H—SUGARS; DERIVATIVES THEREOF; NUCLEOSIDES; NUCLEOTIDES; NUCLEIC ACIDS
- C07H17/00—Compounds containing heterocyclic radicals directly attached to hetero atoms of saccharide radicals
- C07H17/04—Heterocyclic radicals containing only oxygen as ring hetero atoms
- C07H17/08—Hetero rings containing eight or more ring members, e.g. erythromycins
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N15/00—Mutation or genetic engineering; DNA or RNA concerning genetic engineering, vectors, e.g. plasmids, or their isolation, preparation or purification; Use of hosts therefor
- C12N15/09—Recombinant DNA-technology
- C12N15/11—DNA or RNA fragments; Modified forms thereof; Non-coding nucleic acids having a biological activity
- C12N15/52—Genes encoding for enzymes or proenzymes
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N9/00—Enzymes; Proenzymes; Compositions thereof; Processes for preparing, activating, inhibiting, separating or purifying enzymes
- C12N9/88—Lyases (4.)
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12P—FERMENTATION OR ENZYME-USING PROCESSES TO SYNTHESISE A DESIRED CHEMICAL COMPOUND OR COMPOSITION OR TO SEPARATE OPTICAL ISOMERS FROM A RACEMIC MIXTURE
- C12P17/00—Preparation of heterocyclic carbon compounds with only O, N, S, Se or Te as ring hetero atoms
- C12P17/18—Preparation of heterocyclic carbon compounds with only O, N, S, Se or Te as ring hetero atoms containing at least two hetero rings condensed among themselves or condensed with a common carbocyclic ring system, e.g. rifamycin
- C12P17/182—Heterocyclic compounds containing nitrogen atoms as the only ring heteroatoms in the condensed system
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12P—FERMENTATION OR ENZYME-USING PROCESSES TO SYNTHESISE A DESIRED CHEMICAL COMPOUND OR COMPOSITION OR TO SEPARATE OPTICAL ISOMERS FROM A RACEMIC MIXTURE
- C12P19/00—Preparation of compounds containing saccharide radicals
- C12P19/44—Preparation of O-glycosides, e.g. glucosides
- C12P19/60—Preparation of O-glycosides, e.g. glucosides having an oxygen of the saccharide radical directly bound to a non-saccharide heterocyclic ring or a condensed ring system containing a non-saccharide heterocyclic ring, e.g. coumermycin, novobiocin
- C12P19/62—Preparation of O-glycosides, e.g. glucosides having an oxygen of the saccharide radical directly bound to a non-saccharide heterocyclic ring or a condensed ring system containing a non-saccharide heterocyclic ring, e.g. coumermycin, novobiocin the hetero ring having eight or more ring members and only oxygen as ring hetero atoms, e.g. erythromycin, spiramycin, nystatin
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07K—PEPTIDES
- C07K2319/00—Fusion polypeptide
Definitions
- the present invention relates to polyketides and their synthesis and use. It is particularly, but not exclusively, concerned with variants of rapamycin.
- Rapamycin (see Fig. 2) is a lipophilic macrolide, of molecular weight 914, with a 1, 2, 3-tricarbonyl moiety linked to a pipecolic acid lactone . Sequencing of the putative biosynthetic genes of rapamycin has revealed the presence of three exceptionally large open reading frames encoding the modular polyketide synthase (Schwecke et al., P.N.A.S. 92 (17) 7839-7843 (1995)). On either side of these very large genes are ranged open reading frames which appear to encode enzymes that would be required for rapamycin biosynthesis.
- the cluster also contains a novel gene (rapL) whose product is proposed to catalyse the formation of the rapamycin precursor L-pipecolate (2) through the cyclodeamination of L-lysine (1) (Molnar et al., Gene 169, 1-7 (1996) ) :-
- rapL novel gene
- rapamycin requires a modular polyketide synthase, which uses a shikimate-derived starter unit and which carries out a total of fourteen successive cycles of polyketide chain elongation that resemble the steps in fatty acid biosynthesis.
- L- pipecolic acid is then incorporated into the chain, followed by closure of the macrocyclic ring, and both these steps are believed to be catalyzed by a pipecolate- incorporating enzyme (PIE), the product of the rapP gene. Further site-specific oxidations and O-methylation steps are then required to produce rapamycin.
- PIE pipecolate- incorporating enzyme
- the organism cannot produce rapamycin under normal growth conditions but can do so if fed pipecolate. Furthermore feeding the mutant organism with different substrates leads to the production of variants of rapamycin.
- the same general method can be applied to other systems which involve a precursor compound which is produced by a gene product, e.g. the very closely related FK506 and immunomycin systems which also " involve pipecolate.
- a process of modifying a gene cluster involved in the biosynthesis of a polyketide said gene cluster including a gene ("the precursor gene") responsible for the production of an enzyme which is responsible for the production of a precursor compound which is incorporated into said polyketide; said process comprising the step of deleting or inactivating said precursor gene.
- said process of deleting or inactivating said precursor gene employs phage-mediated gene replacement .
- the gene cluster is the gene cluster for the production of rapamycin in S . hycrroscopicus and the precursor gene is the rapL gene whose product is responsible for the production of L-pipecolate .
- the invention provides a process for producing a polyketide comprising modifying a gene cluster by the process according to the first aspect and expressing the modified gene cluster in the presence of a variant precursor compound which is incorporated so that a variant polyketide is produced.
- a variant precursor compound for the rapamycin system, examples include L-proline, L-trans-4-hydroxyproline, L-cis-4- hydroxyproline, L-cis-3-hydroxyproline and trans-3-aza- bicyclo [3,1,0] hexane-2-carboxylic acid.
- the invention provides polyketides as producible by the above method, pharmaceuticals comprising such polyketides, and the use of such polyketides in preparing pharmaceutical compositions, e.g. immunosuppressants containing rapamycin variants.
- Fig. 1 shows a portion of the rapamycin gene cluster, wild type and mutated, and the phage vector used to perform mutation;
- Fig. 2 shows structures of rapamycin and some variants
- Figs. 3 and 4 illustrate the effects of rapamycin and variants on human lymphoblastoid cell lines.
- a chromosomal mutant of S . hycrroscopicus was created by phage ⁇ C31-mediated gene replacement using the method of Lomovskaya et al . [Microbiology (UK) 1997, 143 , 815-883] .
- a unique BamH I site was found 42bp into the rapL gene (1032bp long) . This BamH I site was removed by end-filling with E.coli DNA polymerase I thus creating a frameshift in the rapL gene.
- Escherichia coli DH10B Bacterial strains, phages and growth conditions Escherichia coli DH10B (GibcoBRL) was grown in
- the rapamycin producer Streptomyces hygroscopicus NRRL 5491 (from ATCC) and its derivatives were maintained on SY agar (Soluble starch 1.5%; yeast extract 0.1%; K 2 HP0 4 0.1%; MgS0 4 x 7 H 2 0 0.1%; NaCl 0.3%; N-tri [Hydroxymethyl] methyl-2- aminoethanesulfonic acid (Tes) buffer 30 mM, pH7.4 ; agar 1.5%), and cultivated in Tryptic Soy Broth with 1.0% glucose, lOOmM MES pH6.0 , supplemented with 10 ug/ml viomycin when required.
- SY agar Soluble starch 1.5%; yeast extract 0.1%; K 2 HP0 4 0.1%; MgS0 4 x 7 H 2 0 0.1%; NaCl 0.3%; N-tri [Hydroxymethyl] methyl-2- aminoethanesulfonic acid (Tes) buffer 30
- ⁇ rapL carrying a frameshi-ft in the rapL gene for homologous recombination in S .hygroscopicus pUC3EcoRI was constructed by cloning a 3034bp Eco RI fragment (nucleotides 93956 to 96990 of the rap cluster) (T. Schwecke et al . , P.N.A.S. 92. 7839-7843 (1995)) encompassing the entire rapL gene flanked by rapK and part of the rapM genes respectively into an Eco Rl-cut pUC18 modified vector where the Bam HI site in the polylinker region has been removed.
- a unique Bam HI site (starting at nucleotide 95036 of the rap cluster) was found 42bp into the rapL gene (nucleotide 95078 to 94047 of the rap cluster; 1032bp long) .
- Plasmid pUC3Eco RI was digested with Bam HI and the cohesive ends were filled in by treating it with E. coli DNA polymerase I (Klenow fragment) .
- the ligated plasmid DNA was redigested with Bam HI and used to transform E. coli . Ampicillin resistant transformants were selected and their plasmid DNA checked for the removal of the Bam HI site by restriction enzyme analysis. This was confirmed by DNA sequencing.
- the 3kb insert was excised from the plasmid with Eco RI and the cohesive ends blunt-ended by treatment with E. coli DNA polymerase I (Klenow fragment) .
- the blunt -ended insert was cloned into Pvu II cut phage vector KC515, resulting in ⁇ rapL.
- Protoplasts of S . lividans J11326 were transfected with the phage construct as described by Hopwood et al . (1985) . Recombinant phage was identified using PCR analysis. Infection of S .
- hygroscopicus NRRL 5491 with ⁇ rapL was done according to Lomovskaya ete al (Microbiology, 143, 875-883 (1997)) on DNA plates supplemented with glucose, MgS0 4 and Ca(N0 3 ) . Lysogens were selected by overlaying the plates with 50 ⁇ g ml "1 (final concentration) viomycin 24 h post-infection. Strains that had undergone a second recombination event deleting the integrated phage were identified by selecting viomycin sensitive isolates after three rounds of non-selective growth and sporulation on SY plates. The insertion and subsequent loss of the phage were confirmed by genomic Southern hybridizations.
- Precursor feeding and fermentation of S . hygroscopicus ⁇ RapL Precursor feeding of S .hygroscopicus ⁇ RapL was performed routinely in 500 ml flasks containing 100ml of Tryptic Soy Broth with 1.0% glucose, lOOmM MES pH6.0 , supplemented with the appropriate pipecolic acid analogue, at a final concentration of lmg/ml .
- S . hygroscopicus ⁇ RapL was also cultivated in 2 1 flasks containing 400 ml of chemically-defined media as described by Cheng et al (Appl . Microbiol . Biotechnol . 43 , 1096-1098, (1995)).
- Trans 4-hydroxyproline was added to the medium aseptically to a final concentration of lmg/ml.
- the fermentation was carried out at 28°C for 4 days, with an agitation rate of 500 rpm.
- the cells were harvested and extracted with twice their volume of methanol overnight . Purification and analysis of rapamycin and its derivatives
- Rapamycin induces a specific cell cycle arrest in Gl in the cell line 536, which is a human B lymphocytic line immortalised by Epstein Barr virus infection.
- the potency of each analogue was compared to that of rapamycin using the 536 cells as a bioassay.
- the 536 cells obtained from the human genetic mutant cell repository, Camden, New Jersey, USA) were cultured in Iscoves medium supplemented with 10% fetal calf serum. For bioassay, 536 cells were seeded into 96 well microtitre plates at 10,000 per well in lOO ⁇ l of growth medium.
- Drug stocks of ImM in DMSO were prepared and further dilutions were made to give a constant final concentration of 0.1% DMSO in growth medium.
- Control cultures were treated with 0.1% DMSO in growth medium; experimental cultures received a final concentration of 10 "7 M, 10 "8 M, 10 "9 M or 10 "10 M rapamycin or rapamycin analogue.
- Each culture was set up in triplicate and replicate plates were labelled with l ⁇ Ci tritiated thymidine (Amersham International, specific activity 70Ci/mM) per well for 3h at either Oh; 24h; or 48h incubation with drugs.
- the cultures were harvested onto glass fibre paper to trap the DNA following water lysis; free nucleotides were washed away. Radioactivity incorporated into the filter discs / trapped DNA was counted in a Packard scintillation counter using biodegradable scintillation fluid.
- ⁇ RapL was isolated as described in Materials and Methods.
- This mutation was investigated by Southern blot hybridization using the 3kb EcoRI fragment (93956-96990) to probe Bgl II/Bam HI digested chromosomal DNA. Analysis of the wild-type S . hygroscopicus shows the expected 5.9kb (representing nucleotides 89118-95036) and 2.7kb Bam HI/Bgl II fragments (representing nucleotides 95036-97710) after hybridisation.
- HPLC-ESIMS High pressure liquid chromatograph-electrospray ionization mass spectrometry
- the methanol extracts were combined and concentrated under reduced pressure.
- the aqueous residue was diluted with 500 mL of distilled water, and extracted three times with 500 mL of distilled ethyl acetate.
- the combined ethyl acetate extracts were dried with anhydrous sodium sulphate, and evaporated to dryness .
- the resulting yellow residue was purified by flash column chromatography on a 150 mm x 30 mm (diameter) silica gel column [Merck 60] eluted isocratically witeh a 1:1 (v/v) mixture of acetone/hexane .
- the fractions were analysed by electrospray mass spectrometry. MS-MS and Ms n were used to determine the structure of the new rapamycin in the fractions from the flash silica column.
- the fractions containing trans-3-aza- bicyclo [3.1.0] hexane-2 -carboxylic acid rapamycin were further purified by reversed-phase preparative HPLC on a 250 x 20 mm (diameter) Prodigy 0DS3 column (Phenomenex) using gradient elution starting at 70/30 (v/v) acetonitrile/water rising linearly to 100% acetonitrile over 25 minutes.
- the 2 L fermentation yielded about 4 mg of pure trans-3-aza-bicyclo [3.1.0] hexane-2 -carboxylic acid rapamycin.
- the dose response of human lymphoblastoid cell lines 536 was measured.
- the mean cpm of radiolabelled thymidine incorporated into the untreated controls shows that 0-3 h drug exposure had no appreciable effect on DNA synthesis up to lOOnM of rapamycin, prolylrapamycin, or 4-hydroxy-prolyl-26- demethoxy-rapamycin. This implies that none of the compounds were toxic to the 536 cell line.
- the 536 cells showed a concentration- dependent inhibition of DNA synthesis with an ID50% of InM for rapamycin; and 3nM for prolylrapamycin.
- rapamycin 4- hydroxy-prolyl-26-demethoxy-rapamycin was also inhibitory but did not reach 50% at lOOnM.
- Previous experiments have shown that rapamycin is a profound inhibitor of Gl progression in the 536 cell line (Metcalfe et al . , Oncogene 15, 1635-1642 (1997)). This is also suggested in these experiments for the rapamycin analogues, since no significant effect was found at 3h but inhibition was observed once the cell population had time to proceed through a complete cell cycle (24h) and reach the drug arrest point .
Abstract
Description
Claims
Priority Applications (7)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
AT98924463T ATE307198T1 (en) | 1997-05-28 | 1998-05-28 | POLYKETIDES AND THEIR PRODUCTION AND USE |
DE69831943T DE69831943T2 (en) | 1997-05-28 | 1998-05-28 | POLYETHYDES AND THEIR PREPARATION AND APPLICATION |
EP98924463A EP0983348B1 (en) | 1997-05-28 | 1998-05-28 | Polyketides and their synthesis and use |
AU76661/98A AU7666198A (en) | 1997-05-28 | 1998-05-28 | Polyketides and their synthesis and use |
US10/307,595 US7018808B2 (en) | 1997-05-28 | 2002-12-02 | Polyketides and their synthesis and use |
US11/328,642 US7381546B2 (en) | 1997-05-28 | 2006-01-10 | Polyketides and their synthesis and use |
US12/129,295 US20080287483A1 (en) | 1997-05-28 | 2008-05-29 | Polyketides and Their Synthesis and Use |
Applications Claiming Priority (2)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
GBGB9710962.3A GB9710962D0 (en) | 1997-05-28 | 1997-05-28 | Polyketides and their synthesis |
GB9710962.3 | 1997-05-28 |
Related Child Applications (2)
Application Number | Title | Priority Date | Filing Date |
---|---|---|---|
US09424751 A-371-Of-International | 1998-05-28 | ||
US10/307,595 Continuation US7018808B2 (en) | 1997-05-28 | 2002-12-02 | Polyketides and their synthesis and use |
Publications (2)
Publication Number | Publication Date |
---|---|
WO1998054308A2 true WO1998054308A2 (en) | 1998-12-03 |
WO1998054308A3 WO1998054308A3 (en) | 1999-04-08 |
Family
ID=10813155
Family Applications (1)
Application Number | Title | Priority Date | Filing Date |
---|---|---|---|
PCT/GB1998/001559 WO1998054308A2 (en) | 1997-05-28 | 1998-05-28 | Polyketides and their synthesis and use |
Country Status (8)
Country | Link |
---|---|
US (3) | US7018808B2 (en) |
EP (1) | EP0983348B1 (en) |
AT (1) | ATE307198T1 (en) |
AU (1) | AU7666198A (en) |
DE (1) | DE69831943T2 (en) |
ES (1) | ES2251084T3 (en) |
GB (1) | GB9710962D0 (en) |
WO (1) | WO1998054308A2 (en) |
Cited By (8)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
WO2000047724A2 (en) * | 1999-02-09 | 2000-08-17 | Board Of Trustees Of The Leland Stanford Junior University | Methods to mediate polyketide synthase module effectiveness |
WO2001034816A1 (en) * | 1999-10-29 | 2001-05-17 | Kosan Biosciences, Inc. | Rapamycin analogs |
US6495348B1 (en) | 1993-10-07 | 2002-12-17 | Regents Of The University Of Minnesota | Mitomycin biosynthetic gene cluster |
WO2002101003A2 (en) * | 2001-06-08 | 2002-12-19 | Rhodia Chimie | Stereoselective preparation of cyclic l-amino acids |
WO2005100366A1 (en) * | 2004-04-14 | 2005-10-27 | Wyeth | Proline cci-779 (proline-rapamycin 42-ester with 2,2-bis (hydroxymethyl) propionic acid) and two-step enzymatic synthesis of proline cci-779 and cci-779 using microbial lipase |
WO2006016167A2 (en) * | 2004-08-11 | 2006-02-16 | Biotica Technology Limited | 17-desmethylrapamycin and analogues thereof, methods for their roduction and their use as immunosupressants, anticancer agents, antifungal agents, etc. |
US7001748B2 (en) | 1999-02-09 | 2006-02-21 | The Board Of Trustees Of The Leland Stanford Junior University | Methods of making polyketides using hybrid polyketide synthases |
US7300942B2 (en) | 2002-07-16 | 2007-11-27 | Biotica Technology Limited | Production of polyketides and other natural products |
Families Citing this family (6)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
CN102144961A (en) * | 2003-09-18 | 2011-08-10 | 参天制药株式会社 | Transscleral delivery |
US8663639B2 (en) * | 2005-02-09 | 2014-03-04 | Santen Pharmaceutical Co., Ltd. | Formulations for treating ocular diseases and conditions |
WO2006086750A1 (en) * | 2005-02-09 | 2006-08-17 | Macusight, Inc. | Liquid formulations for treatment of diseases or conditions |
JP5528708B2 (en) | 2006-02-09 | 2014-06-25 | 参天製薬株式会社 | Stable formulations and methods for preparing and using them |
US8222271B2 (en) | 2006-03-23 | 2012-07-17 | Santen Pharmaceutical Co., Ltd. | Formulations and methods for vascular permeability-related diseases or conditions |
US20080265343A1 (en) * | 2007-04-26 | 2008-10-30 | International Business Machines Corporation | Field effect transistor with inverted t shaped gate electrode and methods for fabrication thereof |
Citations (2)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
EP0589703A1 (en) * | 1992-09-24 | 1994-03-30 | American Home Products Corporation | Proline derivative of rapamycin, production and application thereof |
WO1994010843A1 (en) * | 1992-11-10 | 1994-05-26 | Smithkline Beecham Corporation | Rapamycin derivatives |
-
1997
- 1997-05-28 GB GBGB9710962.3A patent/GB9710962D0/en active Pending
-
1998
- 1998-05-28 AU AU76661/98A patent/AU7666198A/en not_active Abandoned
- 1998-05-28 DE DE69831943T patent/DE69831943T2/en not_active Expired - Lifetime
- 1998-05-28 WO PCT/GB1998/001559 patent/WO1998054308A2/en active IP Right Grant
- 1998-05-28 AT AT98924463T patent/ATE307198T1/en active
- 1998-05-28 EP EP98924463A patent/EP0983348B1/en not_active Expired - Lifetime
- 1998-05-28 ES ES98924463T patent/ES2251084T3/en not_active Expired - Lifetime
-
2002
- 2002-12-02 US US10/307,595 patent/US7018808B2/en not_active Expired - Fee Related
-
2006
- 2006-01-10 US US11/328,642 patent/US7381546B2/en not_active Expired - Fee Related
-
2008
- 2008-05-29 US US12/129,295 patent/US20080287483A1/en not_active Abandoned
Patent Citations (2)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
EP0589703A1 (en) * | 1992-09-24 | 1994-03-30 | American Home Products Corporation | Proline derivative of rapamycin, production and application thereof |
WO1994010843A1 (en) * | 1992-11-10 | 1994-05-26 | Smithkline Beecham Corporation | Rapamycin derivatives |
Non-Patent Citations (8)
Cited By (24)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
US6495348B1 (en) | 1993-10-07 | 2002-12-17 | Regents Of The University Of Minnesota | Mitomycin biosynthetic gene cluster |
US7001748B2 (en) | 1999-02-09 | 2006-02-21 | The Board Of Trustees Of The Leland Stanford Junior University | Methods of making polyketides using hybrid polyketide synthases |
WO2000047724A3 (en) * | 1999-02-09 | 2000-12-07 | Univ Leland Stanford Junior | Methods to mediate polyketide synthase module effectiveness |
US6753173B1 (en) | 1999-02-09 | 2004-06-22 | Board Of Trustees Of The Leland Stanford Junior University | Methods to mediate polyketide synthase module effectiveness |
WO2000047724A2 (en) * | 1999-02-09 | 2000-08-17 | Board Of Trustees Of The Leland Stanford Junior University | Methods to mediate polyketide synthase module effectiveness |
WO2001034816A1 (en) * | 1999-10-29 | 2001-05-17 | Kosan Biosciences, Inc. | Rapamycin analogs |
US6670168B1 (en) | 1999-10-29 | 2003-12-30 | Kosan Bioscience, Inc. | Recombinant Streptomyces hygroscopicus host cells that produce 17-desmethylrapamycin |
WO2002101003A2 (en) * | 2001-06-08 | 2002-12-19 | Rhodia Chimie | Stereoselective preparation of cyclic l-amino acids |
WO2002101003A3 (en) * | 2001-06-08 | 2004-02-26 | Rhodia Chimie Sa | Stereoselective preparation of cyclic l-amino acids |
US7425530B2 (en) | 2001-06-08 | 2008-09-16 | Rhodia Chimie | Stereoselective preparation of cyclic L-amino acids |
US7645768B2 (en) | 2002-07-16 | 2010-01-12 | Wyeth | Production of polyketides and other natural products |
EP2277898A2 (en) | 2002-07-16 | 2011-01-26 | Biotica Technology Limited | Rapamycin analogues |
US7300942B2 (en) | 2002-07-16 | 2007-11-27 | Biotica Technology Limited | Production of polyketides and other natural products |
US9605001B2 (en) | 2002-07-16 | 2017-03-28 | Buck Institute For Research On Aging | Production of polyketides and other natural products |
US7390895B2 (en) | 2002-07-16 | 2008-06-24 | Biotica Technology Limited | Production of polyketides and other natural products |
EP1967520A2 (en) | 2002-07-16 | 2008-09-10 | Biotica Technology Limited | Production of polyketides and other natural products |
US9040259B2 (en) | 2002-07-16 | 2015-05-26 | Buck Institute For Research On Aging | Production of polyketides and other natural products |
EP2277898A3 (en) * | 2002-07-16 | 2011-06-01 | Biotica Technology Limited | Rapamycin analogues |
US7202256B2 (en) | 2004-04-14 | 2007-04-10 | Wyeth | Proline CCI-779, production of and uses therefor, and two-step enzymatic synthesis of proline CCI-779 and CCI-779 |
WO2005100366A1 (en) * | 2004-04-14 | 2005-10-27 | Wyeth | Proline cci-779 (proline-rapamycin 42-ester with 2,2-bis (hydroxymethyl) propionic acid) and two-step enzymatic synthesis of proline cci-779 and cci-779 using microbial lipase |
JP2008509895A (en) * | 2004-08-11 | 2008-04-03 | バイオティカ テクノロジー リミテッド | Manufacture of polyketides and other natural products |
US7803808B2 (en) | 2004-08-11 | 2010-09-28 | Wyeth Llc | Production of polyketides and other natural products |
WO2006016167A2 (en) * | 2004-08-11 | 2006-02-16 | Biotica Technology Limited | 17-desmethylrapamycin and analogues thereof, methods for their roduction and their use as immunosupressants, anticancer agents, antifungal agents, etc. |
WO2006016167A3 (en) * | 2004-08-11 | 2006-04-13 | Biotica Tech Ltd | 17-desmethylrapamycin and analogues thereof, methods for their roduction and their use as immunosupressants, anticancer agents, antifungal agents, etc. |
Also Published As
Publication number | Publication date |
---|---|
DE69831943D1 (en) | 2006-03-02 |
US20060105436A1 (en) | 2006-05-18 |
AU7666198A (en) | 1998-12-30 |
US20030104585A1 (en) | 2003-06-05 |
US7381546B2 (en) | 2008-06-03 |
DE69831943T2 (en) | 2006-07-06 |
US20080287483A1 (en) | 2008-11-20 |
WO1998054308A3 (en) | 1999-04-08 |
EP0983348A2 (en) | 2000-03-08 |
ATE307198T1 (en) | 2005-11-15 |
EP0983348B1 (en) | 2005-10-19 |
US7018808B2 (en) | 2006-03-28 |
GB9710962D0 (en) | 1997-07-23 |
ES2251084T3 (en) | 2006-04-16 |
Similar Documents
Publication | Publication Date | Title |
---|---|---|
US7381546B2 (en) | Polyketides and their synthesis and use | |
EP2342335B1 (en) | Novel gene cluster | |
Bibb et al. | Analysis of the nucleotide sequence of the Streptomyces glaucescens tcmI genes provides key information about the enzymology of polyketide antibiotic biosynthesis. | |
Ward et al. | Chalcomycin biosynthesis gene cluster from Streptomyces bikiniensis: novel features of an unusual ketolide produced through expression of the chm polyketide synthase in Streptomyces fradiae | |
JP2000515390A (en) | Novel polyketide derivative and recombinant method for producing the same | |
JP2000511063A (en) | Polyketides and their synthesis | |
EP0929681B1 (en) | Rifamycin biosynthesis gene cluster | |
WO2005121327A2 (en) | Biosynthetic gene cluster for the production of a complex polyketide | |
Arisawa et al. | Direct fermentative production of acyltylosins by genetically-engineered strains of Streptomyces fradiae | |
JP2008278895A (en) | Biosynthetic gene for producing butenyl-spinosyn insecticide | |
WILSON et al. | Molecular analysis of tlrB, an antibiotic-resistance gene from tylosin-producing Streptomyces fradiae, and discovery of a novel resistance mechanism | |
CN110305881B (en) | Biosynthetic gene cluster of polyketide neoenterocins and application thereof | |
EP1632569B1 (en) | Strain belonging to the genus streptomyces and being capable of producing nemadictin and process for producing nemadictin using the strain | |
Tornus et al. | Identification of four genes from the granaticin biosynthetic gene cluster of Streptomyces violaceoruber Tü22 involved in the biosynthesis of L-rhodinose | |
Novakova et al. | Cloning and characterization of a new polyketide synthase gene cluster in Streptomyces aureofaciens CCM 3239 | |
Paulus et al. | New Alpiniamides From sp. IB2014/011-12 Assembled by an Unusual Hybrid Non-ribosomal Peptide Synthetase-AT Polyketide Synthase Enzyme. | |
SOCKALINGAM | Genetic studies on a soil streptomyces sp. that produces an antifungal compoud | |
JP2009502187A (en) | Genes involved in thiocoralin biosynthesis and their heterologous production | |
Praseuth et al. | APPLIED CELLULAR PHYSIOLOGY AND METABOLIC ENGINEERING | |
Virolle et al. | Constanze Paulus1, 2, Yuriy Rebets 2, Josef Zapp2, Christian Rückert 3, Jörn Kalinowski 3 and Andriy Luzhetskyy | |
SI20274A (en) | Novel polyketides, anthrone derivatives | |
KR20140021083A (en) | Two-component system genes derived from streptomyces acidiscabies atcc 49003 and the production method of antibiotics using the same genes | |
AU2002305118A1 (en) | Biosynthetic genes for butenyl-spinosyn insecticide production |
Legal Events
Date | Code | Title | Description |
---|---|---|---|
AK | Designated states |
Kind code of ref document: A2 Designated state(s): AL AM AT AU AZ BA BB BG BR BY CA CH CN CU CZ DE DK EE ES FI GB GE GH GM GW HU ID IL IS JP KE KG KP KR KZ LC LK LR LS LT LU LV MD MG MK MN MW MX NO NZ PL PT RO RU SD SE SG SI SK SL TJ TM TR TT UA UG US UZ VN YU ZW |
|
AL | Designated countries for regional patents |
Kind code of ref document: A2 Designated state(s): GH GM KE LS MW SD SZ UG ZW AM AZ BY KG KZ MD RU TJ TM AT BE CH CY DE DK ES FI FR GB GR IE IT LU MC NL PT SE BF BJ CF CG CI CM GA GN ML MR NE SN TD TG |
|
DFPE | Request for preliminary examination filed prior to expiration of 19th month from priority date (pct application filed before 20040101) | ||
AK | Designated states |
Kind code of ref document: A3 Designated state(s): AL AM AT AU AZ BA BB BG BR BY CA CH CN CU CZ DE DK EE ES FI GB GE GH GM GW HU ID IL IS JP KE KG KP KR KZ LC LK LR LS LT LU LV MD MG MK MN MW MX NO NZ PL PT RO RU SD SE SG SI SK SL TJ TM TR TT UA UG US UZ VN YU ZW |
|
AL | Designated countries for regional patents |
Kind code of ref document: A3 Designated state(s): GH GM KE LS MW SD SZ UG ZW AM AZ BY KG KZ MD RU TJ TM AT BE CH CY DE DK ES FI FR GB GR IE IT LU MC NL PT SE BF BJ CF CG CI CM GA GN ML MR NE SN TD TG |
|
121 | Ep: the epo has been informed by wipo that ep was designated in this application | ||
WWE | Wipo information: entry into national phase |
Ref document number: 09424751 Country of ref document: US |
|
WWE | Wipo information: entry into national phase |
Ref document number: 1998924463 Country of ref document: EP |
|
NENP | Non-entry into the national phase |
Ref country code: JP Ref document number: 1999500383 Format of ref document f/p: F |
|
WWP | Wipo information: published in national office |
Ref document number: 1998924463 Country of ref document: EP |
|
REG | Reference to national code |
Ref country code: DE Ref legal event code: 8642 |
|
NENP | Non-entry into the national phase |
Ref country code: CA |
|
WWG | Wipo information: grant in national office |
Ref document number: 1998924463 Country of ref document: EP |