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WO1996034941A1 - Utilisation de la protease de cysteine extracellulaire pour inhiber la proliferation des cellules - Google Patents

Utilisation de la protease de cysteine extracellulaire pour inhiber la proliferation des cellules Download PDF

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Publication number
WO1996034941A1
WO1996034941A1 PCT/US1996/005997 US9605997W WO9634941A1 WO 1996034941 A1 WO1996034941 A1 WO 1996034941A1 US 9605997 W US9605997 W US 9605997W WO 9634941 A1 WO9634941 A1 WO 9634941A1
Authority
WO
WIPO (PCT)
Prior art keywords
cysteine protease
protease
cells
proliferation
neoplastic cells
Prior art date
Application number
PCT/US1996/005997
Other languages
English (en)
Inventor
James M. Musser
Vivek Kapur
Honnavara N. Ananthaswamy
Antonio Fernandez
Original Assignee
Baylor College Of Medicine
Board Of Regents, The University Of Texas System
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 Baylor College Of Medicine, Board Of Regents, The University Of Texas System filed Critical Baylor College Of Medicine
Priority to AU57188/96A priority Critical patent/AU5718896A/en
Publication of WO1996034941A1 publication Critical patent/WO1996034941A1/fr

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Classifications

    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K38/00—Medicinal preparations containing peptides
    • A61K38/16—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
    • A61K38/43—Enzymes; Proenzymes; Derivatives thereof
    • A61K38/46—Hydrolases (3)
    • A61K38/48—Hydrolases (3) acting on peptide bonds (3.4)
    • A61K38/4873—Cysteine endopeptidases (3.4.22), e.g. stem bromelain, papain, ficin, cathepsin H
    • 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)
    • C12N9/52—Proteinases, e.g. Endopeptidases (3.4.21-3.4.25) derived from bacteria or Archaea

Definitions

  • Metastasis is a process by which tumor cells spread from a tumor to other parts of the body.
  • An antimetastasis therapy is a therapy that reduces or eliminates metastases, or can limit metastases.
  • methods for evaluating the sensitivity of target cells to Streptococcus extracellular cysteine protease include detection of binding of a Streptococcus cysteine protease to the extracellular matrix and/or detection of proteolytic products generated following hydrolysis of protein components of the extracellular matrix (ECM).
  • the cysteine protease retains both the catalytic and binding activities of the native cysteine protease.
  • derived from it is meant that the cysteine protease has the amino acid sequence of all or part of a cysteine protease obtained from S. pyogenes.
  • a microorganism is considered to be the same as or equivalent to Streptococcus pyogenes if in its genome is a coding sequence for an extracellular cysteine protease which is involved in the pathogenesis of the microorganism.
  • the sequence can be naturally occurring, or partially or wholly synthetic.
  • the sequence can be truncated as compared to the native protease, or can include additional sequences at the amino and/or carboxy terminals of the protease.
  • polypeptides employed in the subject invention need not be identical to a cysteine protease obtainable from S. pyogenes, so long as the subject compounds are able to provide for enzymatic interaction with a substrate of a cysteine protease obtainable from at least one of the strains of S. pyogenes.
  • the nucleotide sequence to which the polypeptides "correspond” may exhibit strain-to-strain variation and species-to-species variation.
  • the polypeptides can exhibit various changes such as insertions, deletions, and substitutions, either conservative or non-conservative, where such changes might provide for certain advantages in their use.
  • an ELISA is used to examine the enzymatic activity of each peptide using as substrate component proteins of the ECM.
  • the peptides then are further screened for antiproliferative activity against target hyperproliferative cells, both in vitro and in vivo.
  • the antimetastasis therapy increases survival times by at least about 30% over the survival times typical for the particular type of cancer in the absence of an antimetastasis therapy. More preferably, survival times are increased by at least about 50%, and most preferably by at least about 70%.
  • one or more transforming growth factors may be employed, as well as one or more interferons or tumor necrosis factor may be employed.
  • active fragments can be employed, as well as chimeras in which a portion of one related molecule is joined to a portion of another related molecule.
  • the composition will vary widely depending upon its intended purpose, the desired ratio of the components, as well as the nature of the components and their activity. Thus, depending upon the nature of the malignancy, its response to the different components and their synergistic activity, the ratios of the various components will vary. For the most part, the amount of cysteine protease will generally be in the range of about 0.5 to 25 % of the amount of Oncostatin M or congener employed by itself.
  • the zymogen form can be purified with a closely similar protocol, except cysteine is omitted from the medium and the culture is incubated in the absence of supplemental CO 2 .
  • thermocycling parameters were: denaturation at 94°C for 1 minute, annealing at 55°C for 2 minutes, and extension at 72°C for 2.5 minutes for a total of 30 cycles. A final extension at 72°C for 15 minutes was used.
  • a linker with two copies of the recognition sequence for the enzyme SapI (GCTCTTC) is used to create six base deletions (three bases on each end of the linker), or random two amino acid deletions, across the speB gene. Flanking bases are also randomized by filling the ends of the target sequence after linker excision, then inserting a second blunt end linker that includes a random sequence in place of the Ns. The second linker is then removed by digestion with Sapl and the target sequence is ligated to generate substitutions.
  • the proteolytically inactive precursor of the streptococcal protease is thought to be a member of the family of superantigenic streptococcal and staphylococcal exotoxins (Marrack and Kappler, Science, 248:705-11 (1990); Tomai et al, Infec. Immun., 60:701-05 (1992), Abe et al., J. Immunol., 146:3747-50 (1991). Since superantigens stimulate T-cells to release tumor necrosis factor (Miethke et al., J. Exp. Med., 175:91-98

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  • Health & Medical Sciences (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • Chemical & Material Sciences (AREA)
  • Engineering & Computer Science (AREA)
  • Bioinformatics & Cheminformatics (AREA)
  • Zoology (AREA)
  • Organic Chemistry (AREA)
  • General Health & Medical Sciences (AREA)
  • Medicinal Chemistry (AREA)
  • Genetics & Genomics (AREA)
  • Wood Science & Technology (AREA)
  • Gastroenterology & Hepatology (AREA)
  • Epidemiology (AREA)
  • Biochemistry (AREA)
  • General Engineering & Computer Science (AREA)
  • Biomedical Technology (AREA)
  • Molecular Biology (AREA)
  • Biotechnology (AREA)
  • Pharmacology & Pharmacy (AREA)
  • Microbiology (AREA)
  • Proteomics, Peptides & Aminoacids (AREA)
  • Immunology (AREA)
  • Animal Behavior & Ethology (AREA)
  • Public Health (AREA)
  • Veterinary Medicine (AREA)
  • Medicines That Contain Protein Lipid Enzymes And Other Medicines (AREA)

Abstract

La présente invention concerne des compositions comprenant une protéase de cystéine provenant par exemple d'espèces streptococciques. Ces compositions trouvent une application dans la modulation de la croisance des cellules, notamment en tant qu'agent inhibiteur de la prolifération cellulaire, et plus particulièrement des cellules tumorales. Les compositions inhibant la croissance cellulaire peuvent également comprendre un adjuvant. Des méthodes de criblage pour identifier les cellules tumorales sensibles aux effets de modulation de la croissance de la cystéine-protéase sont également décrites.
PCT/US1996/005997 1995-05-01 1996-04-30 Utilisation de la protease de cysteine extracellulaire pour inhiber la proliferation des cellules WO1996034941A1 (fr)

Priority Applications (1)

Application Number Priority Date Filing Date Title
AU57188/96A AU5718896A (en) 1995-05-01 1996-04-30 Use of extracellular cysteine protease to inhibit cell proli feration

Applications Claiming Priority (2)

Application Number Priority Date Filing Date Title
US08/432,692 1995-05-01
US43269295A 1995-05-09 1995-05-09

Publications (1)

Publication Number Publication Date
WO1996034941A1 true WO1996034941A1 (fr) 1996-11-07

Family

ID=23717219

Family Applications (1)

Application Number Title Priority Date Filing Date
PCT/US1996/005997 WO1996034941A1 (fr) 1995-05-01 1996-04-30 Utilisation de la protease de cysteine extracellulaire pour inhiber la proliferation des cellules

Country Status (2)

Country Link
AU (1) AU5718896A (fr)
WO (1) WO1996034941A1 (fr)

Cited By (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US20090162405A1 (en) * 2006-12-14 2009-06-25 Yong Qian Proteinase-engineered cancer vaccine induces immune responses to prevent cancer and to systemically kill cancer cells
US7750132B2 (en) * 1997-06-25 2010-07-06 The United States Of America As Represented By The Secretary Of The Navy Altered superantigen toxins
US20150132285A1 (en) * 2005-10-03 2015-05-14 Yong Qian Proteinase-engineered Cancer Vaccine Induces Immune Responses to Prevent Cancer and to Systemically Kill Cancer Cells

Citations (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US4844897A (en) * 1985-09-13 1989-07-04 Hiroshi Maeda Anti-tumor protease preparations

Patent Citations (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US4844897A (en) * 1985-09-13 1989-07-04 Hiroshi Maeda Anti-tumor protease preparations

Cited By (4)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US7750132B2 (en) * 1997-06-25 2010-07-06 The United States Of America As Represented By The Secretary Of The Navy Altered superantigen toxins
US20150132285A1 (en) * 2005-10-03 2015-05-14 Yong Qian Proteinase-engineered Cancer Vaccine Induces Immune Responses to Prevent Cancer and to Systemically Kill Cancer Cells
US9669082B2 (en) * 2005-10-03 2017-06-06 Yong Qian Proteinase-engineered cancer vaccine induces immune responses to prevent cancer and to systemically kill cancer cells
US20090162405A1 (en) * 2006-12-14 2009-06-25 Yong Qian Proteinase-engineered cancer vaccine induces immune responses to prevent cancer and to systemically kill cancer cells

Also Published As

Publication number Publication date
AU5718896A (en) 1996-11-21

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