WO2000074711A2 - 10-formyltetrahydrofolate dehydrogenase as therapeutical agent - Google Patents
10-formyltetrahydrofolate dehydrogenase as therapeutical agent Download PDFInfo
- Publication number
- WO2000074711A2 WO2000074711A2 PCT/EP2000/005066 EP0005066W WO0074711A2 WO 2000074711 A2 WO2000074711 A2 WO 2000074711A2 EP 0005066 W EP0005066 W EP 0005066W WO 0074711 A2 WO0074711 A2 WO 0074711A2
- Authority
- WO
- WIPO (PCT)
- Prior art keywords
- formyltetrahydrofolate dehydrogenase
- therapeutical agent
- peg
- cone
- formyltetrahydrofolate
- Prior art date
Links
Classifications
-
- 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
- C12N9/0004—Oxidoreductases (1.)
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P35/00—Antineoplastic agents
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12Y—ENZYMES
- C12Y105/00—Oxidoreductases acting on the CH-NH group of donors (1.5)
- C12Y105/01—Oxidoreductases acting on the CH-NH group of donors (1.5) with NAD+ or NADP+ as acceptor (1.5.1)
- C12Y105/01006—Formyltetrahydrofolate dehydrogenase (1.5.1.6)
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K38/00—Medicinal preparations containing peptides
Definitions
- the present invention relates to the use of 10- formyltetrahydrofolate dehydrogenase as therapeutical agent, in particular as cytotoxic and antitumour agent.
- 10-Formyltetrahydrofolate dehydrogenase is an enzyme present in the liver and in the nervous system of mammals. No therapeutical use for such enzyme has been disclosed up to now.
- cDNA from rat 10-formyltetrahydrofolate dehydrogenase has been disclosed in J. Biol . Chem. 266(8), 4965-4973, 1991, while cDNA of the same human enzyme has been disclosed more recently (Biochem. Mol . Biol. Int., 47(3), 407-415, 1999) .
- mammal 10- formyltetrahydrofolate dehydrogenase is capable of inducing a marked cytotoxic response against tumour cells, when administered to tumour-bearing patients or animals. This cytotoxicity seems to be mediated by cytotoxic antibodies to human tumour cells, particularly carcinomas and adenocarcinomas .
- Cytotoxicity can be quantified in vitro on Jurkat and Kato III cells using conventional methods, based for example on the use of commercial kits such as the CDC-UK kit (Pharmaproduct) .
- the appearance of cytotoxicity in rabbits serum was observed already after a first treatment with the enzyme (1 mg/animal in saline solution) on Jurkat and Kato III cells. Therefore, the invention also relates to pharmaceutical compositions containing as active ingredient an effective dose of 10 -formyltetrahydrofolate dehydrogenase .
- compositions of the invention will be administered to tumour patients using the conventional administration routes for proteins and polypeptides, for example the subcutaneous or intramuscular routes .
- the treatment may be repeated, a treatment comprising one-two week separated administrations of doses ranging from 0.1 to 20 mg of enzyme being preferred.
- very low dosages such as 1.10 - 1.10 g
- concentrations of active ingredient ranging from 10 "6 to 10 ⁇ 10 M.
- 10-Formyltetrahydrofolate dehydrogenase can be prepared by conventional recombinant DNA methods or it can be extracted from the liver of animals, for example from liver of bovine, ovine or swine. Goat liver proved to be a particularly abundant source of this enzyme.
- the extraction process comprises the treatment of livers with solutions buffered at pH 7.4 (PBS) followed by precipitation with 15% polyethylene glycol 6000, chromatography on TSK-DEAE or DEAE-Sephacell at pH 8, elution with 0.3 M NaCl and purification on TSK SW3000.
- PBS pH 7.4
- the pellet is taken up into 61 ml of 0.03M Tris/HCl pH 8, whereas the supernatant (340 ml) is reprecipitated with 5% PEG 6000 (17 g) , then 10% PEG 6000 stirring for 1 hour at 4°C.
- the supernatant (345 ml) is treated with 5% PEG 6000 (17.25 g) , then again with 5% PEG 6000, stirring for 1 hour at 4°C, then centrifuged on J6 at 4,000 g for 30 A
- Leg 2 200 ml 0.03M Tris/HCl pH 8 + 0.3M NaCl for 30 minutes at r.t. + 200 ml of washing.
- Leg 3 200 ml 0.03M Tris/HCl pH 8 + 1M NaCl for 30 minutes at r.t. + 200 ml of washing.
- LEG 3 volume about 400 ml cone: 137 //ml.
- LEG 2 is cone, on PM 30 to an about 20 ml final volume, concentration of about 3.6 mg/ml.
- LEG 2 from DEAE-S obtained above is purified in prep.
Landscapes
- Chemical & Material Sciences (AREA)
- Health & Medical Sciences (AREA)
- Organic Chemistry (AREA)
- Life Sciences & Earth Sciences (AREA)
- Genetics & Genomics (AREA)
- Engineering & Computer Science (AREA)
- Bioinformatics & Cheminformatics (AREA)
- General Health & Medical Sciences (AREA)
- Wood Science & Technology (AREA)
- Zoology (AREA)
- Medicinal Chemistry (AREA)
- Biochemistry (AREA)
- General Engineering & Computer Science (AREA)
- Biomedical Technology (AREA)
- Public Health (AREA)
- Biotechnology (AREA)
- Molecular Biology (AREA)
- Chemical Kinetics & Catalysis (AREA)
- General Chemical & Material Sciences (AREA)
- Nuclear Medicine, Radiotherapy & Molecular Imaging (AREA)
- Pharmacology & Pharmacy (AREA)
- Animal Behavior & Ethology (AREA)
- Microbiology (AREA)
- Veterinary Medicine (AREA)
- Enzymes And Modification Thereof (AREA)
- Medicines That Contain Protein Lipid Enzymes And Other Medicines (AREA)
- Nitrogen Condensed Heterocyclic Rings (AREA)
- Pharmaceuticals Containing Other Organic And Inorganic Compounds (AREA)
- Acyclic And Carbocyclic Compounds In Medicinal Compositions (AREA)
- Saccharide Compounds (AREA)
Abstract
Description
Claims
Priority Applications (5)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
AU59695/00A AU5969500A (en) | 1999-06-03 | 2000-06-02 | 10-formyltetrahydrofolate dehydrogenase as therapeutical agent |
DE60004819T DE60004819T2 (en) | 1999-06-03 | 2000-06-02 | 10-FORMYLTETRAHYDROFOLATE DEHYDROGENASE AS A THERAPEUTIC AGENT |
JP2001501245A JP2003501398A (en) | 1999-06-03 | 2000-06-02 | 10-formyltetrahydrofolate dehydrogenase as therapeutic agent |
AT00945698T ATE247976T1 (en) | 1999-06-03 | 2000-06-02 | 10-FORMYLTETRAHYDROFOLATE DEHYDROGENASE AS A THERAPEUTIC AGENT |
EP00945698A EP1181046B1 (en) | 1999-06-03 | 2000-06-02 | 10-formyltetrahydrofolate dehydrogenase as therapeutical agent |
Applications Claiming Priority (4)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
IT1999MI001243 IT1312341B1 (en) | 1999-06-03 | 1999-06-03 | 10-Formyltetrahydrofolate dehydrogenase, useful as a therapeutic agent, particularly as an antitumor agent possessing high cytotoxicity |
ITMI99A001243 | 1999-06-03 | ||
IT99MI002197 IT1314197B1 (en) | 1999-10-20 | 1999-10-20 | 10-Formyltetrahydrofolate dehydrogenase, useful as a therapeutic agent, particularly as an antitumor agent possessing high cytotoxicity |
ITMI99A002197 | 1999-10-20 |
Publications (2)
Publication Number | Publication Date |
---|---|
WO2000074711A2 true WO2000074711A2 (en) | 2000-12-14 |
WO2000074711A3 WO2000074711A3 (en) | 2001-07-26 |
Family
ID=26331660
Family Applications (1)
Application Number | Title | Priority Date | Filing Date |
---|---|---|---|
PCT/EP2000/005066 WO2000074711A2 (en) | 1999-06-03 | 2000-06-02 | 10-formyltetrahydrofolate dehydrogenase as therapeutical agent |
Country Status (7)
Country | Link |
---|---|
EP (1) | EP1181046B1 (en) |
JP (1) | JP2003501398A (en) |
AT (1) | ATE247976T1 (en) |
AU (1) | AU5969500A (en) |
DE (1) | DE60004819T2 (en) |
ES (1) | ES2204649T3 (en) |
WO (1) | WO2000074711A2 (en) |
Cited By (1)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
CN115166067A (en) * | 2022-05-26 | 2022-10-11 | 中国农业科学院北京畜牧兽医研究所 | Peptide fragment composition for relatively quantitatively analyzing 10-formyltetrahydrofolate dehydrogenase (ALDH) 1L1 of pig and application thereof |
Citations (1)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
WO2000073330A2 (en) * | 1999-06-01 | 2000-12-07 | Proteome Sciences Plc | Method and compositions relating to insulin resistance disorders |
-
2000
- 2000-06-02 AT AT00945698T patent/ATE247976T1/en not_active IP Right Cessation
- 2000-06-02 WO PCT/EP2000/005066 patent/WO2000074711A2/en active IP Right Grant
- 2000-06-02 EP EP00945698A patent/EP1181046B1/en not_active Expired - Lifetime
- 2000-06-02 ES ES00945698T patent/ES2204649T3/en not_active Expired - Lifetime
- 2000-06-02 JP JP2001501245A patent/JP2003501398A/en active Pending
- 2000-06-02 DE DE60004819T patent/DE60004819T2/en not_active Expired - Fee Related
- 2000-06-02 AU AU59695/00A patent/AU5969500A/en not_active Abandoned
Patent Citations (1)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
WO2000073330A2 (en) * | 1999-06-01 | 2000-12-07 | Proteome Sciences Plc | Method and compositions relating to insulin resistance disorders |
Non-Patent Citations (1)
Title |
---|
R.J. COOK ET AL.: "ISOLATION AND CHARACTERIZATION OF cDNA CLONES FOR RAT LIVER 10-FORMYLTETRAHYDROFOLATE DEHYDROGENASE" JOURNAL OF BIOLOGICAL CHEMISTRY, vol. 266, no. 8, 15 March 1991 (1991-03-15), pages 4965-4973, XP002157451 BALTIMORE, MD, US cited in the application * |
Cited By (2)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
CN115166067A (en) * | 2022-05-26 | 2022-10-11 | 中国农业科学院北京畜牧兽医研究所 | Peptide fragment composition for relatively quantitatively analyzing 10-formyltetrahydrofolate dehydrogenase (ALDH) 1L1 of pig and application thereof |
CN115166067B (en) * | 2022-05-26 | 2023-12-26 | 中国农业科学院北京畜牧兽医研究所 | Peptide fragment composition for relatively quantitatively analyzing pig 10-formyl tetrahydrofolate dehydrogenase ALDH1L1 and application |
Also Published As
Publication number | Publication date |
---|---|
WO2000074711A3 (en) | 2001-07-26 |
JP2003501398A (en) | 2003-01-14 |
EP1181046B1 (en) | 2003-08-27 |
EP1181046A2 (en) | 2002-02-27 |
DE60004819T2 (en) | 2004-07-01 |
DE60004819D1 (en) | 2003-10-02 |
ATE247976T1 (en) | 2003-09-15 |
ES2204649T3 (en) | 2004-05-01 |
AU5969500A (en) | 2000-12-28 |
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