WO1991019793A2 - Activateurs de plasminogene hybrides - Google Patents

Activateurs de plasminogene hybrides Download PDF

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Publication number
WO1991019793A2
WO1991019793A2 PCT/GB1991/000945 GB9100945W WO9119793A2 WO 1991019793 A2 WO1991019793 A2 WO 1991019793A2 GB 9100945 W GB9100945 W GB 9100945W WO 9119793 A2 WO9119793 A2 WO 9119793A2
Authority
WO
WIPO (PCT)
Prior art keywords
plasminogen
compound according
hybrid
chain
glu
Prior art date
Application number
PCT/GB1991/000945
Other languages
English (en)
Other versions
WO1991019793A3 (fr
Inventor
Jeffery Hugh Robinson
Original Assignee
Beecham Group Plc
Priority date (The priority date is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the date listed.)
Filing date
Publication date
Application filed by Beecham Group Plc filed Critical Beecham Group Plc
Priority to JP91510746A priority Critical patent/JPH05506363A/ja
Priority to KR1019920703217A priority patent/KR930701607A/ko
Priority to AU80681/91A priority patent/AU648567B2/en
Publication of WO1991019793A2 publication Critical patent/WO1991019793A2/fr
Publication of WO1991019793A3 publication Critical patent/WO1991019793A3/fr

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
    • 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
    • C12N9/14—Hydrolases (3)
    • C12N9/48—Hydrolases (3) acting on peptide bonds (3.4)
    • C12N9/50—Proteinases, e.g. Endopeptidases (3.4.21-3.4.25)
    • C12N9/64—Proteinases, e.g. Endopeptidases (3.4.21-3.4.25) derived from animal tissue
    • C12N9/6421—Proteinases, e.g. Endopeptidases (3.4.21-3.4.25) derived from animal tissue from mammals
    • C12N9/6424—Serine endopeptidases (3.4.21)
    • C12N9/6435—Plasmin (3.4.21.7), i.e. fibrinolysin
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P7/00—Drugs for disorders of the blood or the extracellular fluid
    • A61P7/02—Antithrombotic agents; Anticoagulants; Platelet aggregation inhibitors
    • 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
    • C12N9/14—Hydrolases (3)
    • C12N9/48—Hydrolases (3) acting on peptide bonds (3.4)
    • C12N9/50—Proteinases, e.g. Endopeptidases (3.4.21-3.4.25)
    • C12N9/64—Proteinases, e.g. Endopeptidases (3.4.21-3.4.25) derived from animal tissue
    • C12N9/6421—Proteinases, e.g. Endopeptidases (3.4.21-3.4.25) derived from animal tissue from mammals
    • C12N9/6424—Serine endopeptidases (3.4.21)
    • C12N9/6456—Plasminogen activators
    • C12N9/6459—Plasminogen activators t-plasminogen activator (3.4.21.68), i.e. tPA
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12Y—ENZYMES
    • C12Y304/00—Hydrolases acting on peptide bonds, i.e. peptidases (3.4)
    • C12Y304/21—Serine endopeptidases (3.4.21)
    • C12Y304/21007—Plasmin (3.4.21.7), i.e. fibrinolysin
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12Y—ENZYMES
    • C12Y304/00—Hydrolases acting on peptide bonds, i.e. peptidases (3.4)
    • C12Y304/21—Serine endopeptidases (3.4.21)
    • C12Y304/21069—Protein C activated (3.4.21.69)
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K38/00—Medicinal preparations containing peptides
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K2319/00—Fusion polypeptide

Definitions

  • European Patent No 0009879 discloses derivatives of in vivo fibrinolytic enzymes which are useful therapeutic agents for treating venous thrombosis.
  • the derivatives are characterised by the active catalytic site on the enzymes being blocked by a group which is removable by hydrolysis such that the pseudo first order rate constant for hydrolysis is in the range 10 — Pi to 10 — J - ⁇ sec—1.
  • EP-A-0297882 discloses a hybrid plasminogen activator which comprises the five kringle domains of plasminogen linked to the B-chain of t-PA or u-PA via an amino acid sequence comprising, respectively, the t-PA cleavage site between residues 275 and 276 and the cysteine residue 264 of t-PA or the u-PA cleavage site between residues 158 and 159 and the cysteine residue 148 of u-PA, the catalytic site of the t-PA or u-PA B-chain being optionally blocked by a removable blocking group.
  • 4-methoxybenzoyl plasminogen 1-5 1/t-PA 262-527 including one and two chain variants, lys ⁇ g and glu- ⁇ variants, and mixtures thereof.
  • the invention provides glu- ⁇ , single chain plasminogen 1-541/t-PA 262-527.
  • This variant is preferably present at a level of at least 60%, more preferably 80% relative to the other chain variants.
  • the unacylated hybrid PA is first prepared by expressing DNA encoding the hybrid PA in host cell and recovering the hybrid PA product.
  • the DNA is prepared conventionally by the condensation of appropriate mono-, di- or oligomeric nucleotide units as described in EP-A-0297882.
  • the host cell is prepared conventionally by transformation with a replicable expression vector capable, in the host cell, of expressing the coding DNA.
  • CHO cells were trypsinised and plated out at 5 x 10 * ⁇ per
  • the mixture was buffer-exchanged into 0.02M Tris/0.2M NaCl/0.2M L-arginine/0.01% Tween 80 pH 7.4 using a Sephadex G25 PD10 column.
  • the buffer-exchanged product was stored at -40°C.
  • the deacylation half-life of the product in human, plasma at 37°C was 34 min.

Landscapes

  • Health & Medical Sciences (AREA)
  • Chemical & Material Sciences (AREA)
  • Engineering & Computer Science (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • Genetics & Genomics (AREA)
  • Organic Chemistry (AREA)
  • Bioinformatics & Cheminformatics (AREA)
  • Wood Science & Technology (AREA)
  • Zoology (AREA)
  • Biomedical Technology (AREA)
  • General Health & Medical Sciences (AREA)
  • General Engineering & Computer Science (AREA)
  • Biochemistry (AREA)
  • Biotechnology (AREA)
  • Molecular Biology (AREA)
  • Microbiology (AREA)
  • Medicinal Chemistry (AREA)
  • Pharmacology & Pharmacy (AREA)
  • Public Health (AREA)
  • Chemical Kinetics & Catalysis (AREA)
  • General Chemical & Material Sciences (AREA)
  • Nuclear Medicine, Radiotherapy & Molecular Imaging (AREA)
  • Diabetes (AREA)
  • Animal Behavior & Ethology (AREA)
  • Hematology (AREA)
  • Veterinary Medicine (AREA)
  • Physics & Mathematics (AREA)
  • Biophysics (AREA)
  • Plant Pathology (AREA)
  • Enzymes And Modification Thereof (AREA)
  • Medicines That Contain Protein Lipid Enzymes And Other Medicines (AREA)
  • Fixing For Electrophotography (AREA)
  • Pharmaceuticals Containing Other Organic And Inorganic Compounds (AREA)

Abstract

Enzyme hybride fibrinolytique, préparation de celle-ci, compositions pharmaceutiques contenant ladite enzyme et son utilisation dans le traitement de maladies thrombo-emboliques, en particulier l'infarctus aigu du myocarde.
PCT/GB1991/000945 1990-06-14 1991-06-12 Activateurs de plasminogene hybrides WO1991019793A2 (fr)

Priority Applications (3)

Application Number Priority Date Filing Date Title
JP91510746A JPH05506363A (ja) 1990-06-14 1991-06-12 新規化合物
KR1019920703217A KR930701607A (ko) 1990-06-14 1991-06-12 하이브리드 플라스미노겐 활성제
AU80681/91A AU648567B2 (en) 1990-06-14 1991-06-12 Hybrid plasminogen activators

Applications Claiming Priority (2)

Application Number Priority Date Filing Date Title
GB9013345.5 1990-06-14
GB909013345A GB9013345D0 (en) 1990-06-14 1990-06-14 Novel compounds

Publications (2)

Publication Number Publication Date
WO1991019793A2 true WO1991019793A2 (fr) 1991-12-26
WO1991019793A3 WO1991019793A3 (fr) 1992-01-23

Family

ID=10677652

Family Applications (1)

Application Number Title Priority Date Filing Date
PCT/GB1991/000945 WO1991019793A2 (fr) 1990-06-14 1991-06-12 Activateurs de plasminogene hybrides

Country Status (12)

Country Link
EP (1) EP0535037A1 (fr)
JP (1) JPH05506363A (fr)
KR (1) KR930701607A (fr)
AU (1) AU648567B2 (fr)
CA (1) CA2085224A1 (fr)
GB (1) GB9013345D0 (fr)
IE (1) IE911999A1 (fr)
IL (1) IL98478A0 (fr)
NZ (1) NZ238506A (fr)
PT (1) PT97983A (fr)
WO (1) WO1991019793A2 (fr)
ZA (1) ZA914520B (fr)

Cited By (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2000032759A1 (fr) * 1998-12-02 2000-06-08 Oklahoma Medical Research Foundation Activateur de plasminogene humain

Citations (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
EP0009879A1 (fr) * 1978-09-07 1980-04-16 Beecham Group Plc Dérivés d'enzymes utilisés dans le traitement de thromboses veineuses, compositions les contenant et leur préparations
EP0155387A2 (fr) * 1983-12-24 1985-09-25 Beecham Group Plc Protéine hybride à activité fibrinolytique, son procédé de préparation et composition pharmaceutique
EP0297882A2 (fr) * 1987-07-01 1989-01-04 Beecham Group Plc Activateurs de plasminogène hybrides

Family Cites Families (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
GB9019120D0 (en) * 1990-09-01 1990-10-17 Beecham Group Plc Novel compounds

Patent Citations (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
EP0009879A1 (fr) * 1978-09-07 1980-04-16 Beecham Group Plc Dérivés d'enzymes utilisés dans le traitement de thromboses veineuses, compositions les contenant et leur préparations
EP0155387A2 (fr) * 1983-12-24 1985-09-25 Beecham Group Plc Protéine hybride à activité fibrinolytique, son procédé de préparation et composition pharmaceutique
EP0297882A2 (fr) * 1987-07-01 1989-01-04 Beecham Group Plc Activateurs de plasminogène hybrides

Non-Patent Citations (2)

* Cited by examiner, † Cited by third party
Title
The Biochemical Journal, volume 247, 1987, (GB) R. Cassels et al.: "The interaction of streptokinase-plasminogen activator complex, tissue-type plasminogen activator, urokinase and their acylated derivatives with fibrin and cyanogen bromide digest of fibrinogen", pages 395-400, see abstract; table 2 *
Thrombosis Research, volume 47, no. 6, 15 September 1987, Pergamon Journals Ltd (US) J. St}rzebecher et al.: "Stable acyl-derivatives of tissue-type plasminogen activator", pages 699-703, see table 1; page 701, last sentence - page 702, paragraph 2 *

Cited By (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2000032759A1 (fr) * 1998-12-02 2000-06-08 Oklahoma Medical Research Foundation Activateur de plasminogene humain

Also Published As

Publication number Publication date
IE911999A1 (en) 1991-12-18
AU8068191A (en) 1992-01-07
IL98478A0 (en) 1992-07-15
AU648567B2 (en) 1994-04-28
GB9013345D0 (en) 1990-08-08
KR930701607A (ko) 1993-06-12
ZA914520B (en) 1992-12-30
CA2085224A1 (fr) 1991-12-15
PT97983A (pt) 1992-04-30
EP0535037A1 (fr) 1993-04-07
NZ238506A (en) 1993-07-27
WO1991019793A3 (fr) 1992-01-23
JPH05506363A (ja) 1993-09-22

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