WO1997003194A1 - Analogues polypeptidiques de facteur viii:c a substitution lysine en position 1689 - Google Patents

Analogues polypeptidiques de facteur viii:c a substitution lysine en position 1689 Download PDF

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Publication number
WO1997003194A1
WO1997003194A1 PCT/US1996/011441 US9611441W WO9703194A1 WO 1997003194 A1 WO1997003194 A1 WO 1997003194A1 US 9611441 W US9611441 W US 9611441W WO 9703194 A1 WO9703194 A1 WO 9703194A1
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WO
WIPO (PCT)
Prior art keywords
factor viilc
polypeptide
analog
viilc
nucleic acid
Prior art date
Application number
PCT/US1996/011441
Other languages
English (en)
Inventor
Rae Lyn Burke
Steven Rosenberg
Original Assignee
Chiron Corporation
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 Chiron Corporation filed Critical Chiron Corporation
Priority to AU64560/96A priority Critical patent/AU6456096A/en
Publication of WO1997003194A1 publication Critical patent/WO1997003194A1/fr

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Classifications

    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K14/00—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
    • C07K14/435—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans
    • C07K14/745—Blood coagulation or fibrinolysis factors
    • C07K14/755—Factors VIII, e.g. factor VIII C (AHF), factor VIII Ag (VWF)
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K38/00—Medicinal preparations containing peptides

Definitions

  • a vector suitable for use herein for the production of a recombinant vector comprises a nucleic acid sequence with one or more restriction enzyme recognition sites into which the present nucleic acid molecule of the invention can be inserted. This vector also typically contains a selection marker for detection of the presence of the vector in the host cell.
  • promoters useful for expression of eukaryotic proteins are also suitable.
  • a person skilled in the art would be able to operably ligate such promoters to the present Factor VIILC polypeptide analog coding sequences, for example, as described in Siebenlist et al, Cell (1980) 20: 269, using linkers or adapters to supply any required restriction sites.
  • Promoters for use in bacterial systems also generally will contain a Shine-Dalgarno (SD) sequence operably linked to the DNA encoding the Factor VIILC analog polypeptide.
  • SD Shine-Dalgarno
  • Methods of introducing exogenous DNA into yeast hosts are well known in the art, and typically include either the transformation of spheroplasts or of intact yeast cells treated with alkali cations. Transformations into yeast can be carried out according to the method described in Van Solingen et al. , J. Bact. (1977) 130: 946 and Hsiao et aL , Proc. Natl. Acad. Sci. USA (1979) 76: 3829. However, other methods for introducing DNA into cells such as by nuclear injection, electroporation, or protoplast fusion may also be used as described generally in Sambrook et al. , cited above.
  • gene expression from the polyhedrin promoter occurs at a time when the host cell's ability to process newly synthesized proteins is potentially diminished for certain proteins such as human tissue plasminogen activator.
  • the expression of secretory glycoproteins in BEV systems is complicated due to incomplete secretion of the cloned gene product, thereby trapping the cloned gene product within the cell in an incompletely processed form.
  • the desired DNA sequence can be inserted into the transfer vector, using known techniques.
  • An insect cell host can be cotransformed with the transfer vector containing the inserted desired DNA together with the genomic DNA of wild type baculovirus, usually by cotransfection.
  • the vector and viral genome are allowed to recombine resulting in a recombinant virus that can be easily identified and purified.
  • the packaged recombinant virus can be used to infect insect host cells to express a Factor VIILC polypeptide analog.
  • the recombinant virus can then be isolated and delivered to cells of the subject either in vivo or ex vivo.
  • retroviral systems have been described (U.S. Patent No. 5,219,740; Miller and Rosman, BioTechniques (1989) 7:980-990; Miller, A.D., Human Gene Therapy (1990) 7:5-14; Sca ⁇ a et aL , Virology (1991) 780:849-852; Burns et aL , Proc. Natl. Acad. Sci. USA (1993) 90:8033-8037; and Boris-Lawrie and Temin, Cur. Opin. Genet. Develop. (1993) 3: 102-109.
  • an avipox vector is particularly desirable in human and other mammalian species since members of the avipox genus can only productively replicate in susceptible avian species and therefore are not infective in mammalian cells.
  • Methods for producing recombinant avipoxviruses are known in the art and employ genetic recombination, as described above with respect to the production of vaccinia viruses. See, e.g. , WO 91/12882; WO 89/03429; and WO 92/03545.
  • a medium sample containing wild type heavy and light chains with a total of 37 mU of coagulation activity was incubated in a volume of 200 ⁇ l in 20 M imidazole, 150 mM NaCl, 2.5 mM CaCl 2 , 100 mM lysine HCl, pH 6.8 at 4°C.
  • the reaction was started at time 0 by the addition of either 25 mU of thrombin or buffer alone and transferred to room temperature. Samples were removed at selected times and immediately assayed for coagulation activity. The same experiment was also performed on medium containing the wild type 92 chain and the mutant 80R ⁇ K chain.
  • the 80 ⁇ R, 92 ⁇ R and 92R ⁇ K species were essentially not thrombin activatable under the assay conditions. However the 80R ⁇ K peptide was thrombin activatable to the same extent as the wild type chain although at a slower rate.

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  • Health & Medical Sciences (AREA)
  • Chemical & Material Sciences (AREA)
  • Organic Chemistry (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • Biophysics (AREA)
  • Gastroenterology & Hepatology (AREA)
  • Zoology (AREA)
  • Biochemistry (AREA)
  • Toxicology (AREA)
  • General Health & Medical Sciences (AREA)
  • Genetics & Genomics (AREA)
  • Medicinal Chemistry (AREA)
  • Molecular Biology (AREA)
  • Proteomics, Peptides & Aminoacids (AREA)
  • Hematology (AREA)
  • Micro-Organisms Or Cultivation Processes Thereof (AREA)
  • Preparation Of Compounds By Using Micro-Organisms (AREA)

Abstract

La présente invention se rapporte à des analogues polypeptidiques de Facteur VIII:C qui sont des polypeptides natifs de Facteur VIII:C comportant une substitution de Arg vers Lys au niveau de la position 1689. L'invention décrit également des molécules d'acides nucléiques codant les analogues polypeptidiques de Facteur VIII:C, des vecteurs et des cellules hôtes contenant de telles molécules d'acides nucléiques. L'invention décrit des complexes d'analogues contenant l'analogue. L'invention décrit également des procédés pour produire l'analogue, le complexe d'analogue, les acides nucléiques, les vecteurs et les cellules hôtes, ainsi que des procédés d'utilisation de telles compositions pour la prévention ou le traitement de déficiences en polypeptides actifs de Facteur VIII:C.
PCT/US1996/011441 1995-07-11 1996-07-09 Analogues polypeptidiques de facteur viii:c a substitution lysine en position 1689 WO1997003194A1 (fr)

Priority Applications (1)

Application Number Priority Date Filing Date Title
AU64560/96A AU6456096A (en) 1995-07-11 1996-07-09 Lysine 1689 factor viii:c polypeptide analogs

Applications Claiming Priority (2)

Application Number Priority Date Filing Date Title
US102995P 1995-07-11 1995-07-11
US60/001,029 1995-07-11

Publications (1)

Publication Number Publication Date
WO1997003194A1 true WO1997003194A1 (fr) 1997-01-30

Family

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Family Applications (1)

Application Number Title Priority Date Filing Date
PCT/US1996/011441 WO1997003194A1 (fr) 1995-07-11 1996-07-09 Analogues polypeptidiques de facteur viii:c a substitution lysine en position 1689

Country Status (2)

Country Link
AU (1) AU6456096A (fr)
WO (1) WO1997003194A1 (fr)

Cited By (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US6838437B2 (en) 1996-04-24 2005-01-04 University Of Michigan Inactivation resistant factor VIII
EP2206785A1 (fr) 1998-12-31 2010-07-14 Novartis Vaccines and Diagnostics, Inc. Expression améliorée de polypeptides HIV et production de particules de type virus
US8183344B2 (en) 1996-04-24 2012-05-22 University Of Michigan Inactivation resistant factor VIII

Citations (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO1987007144A1 (fr) * 1986-05-29 1987-12-03 Genetics Institute, Inc. Nouvelles proteines stimulant la coagulation du sang
EP0295597A2 (fr) * 1987-06-19 1988-12-21 BEHRINGWERKE Aktiengesellschaft Molécule similaire au facteur VIII:C, à l'activité coagulante

Patent Citations (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO1987007144A1 (fr) * 1986-05-29 1987-12-03 Genetics Institute, Inc. Nouvelles proteines stimulant la coagulation du sang
EP0295597A2 (fr) * 1987-06-19 1988-12-21 BEHRINGWERKE Aktiengesellschaft Molécule similaire au facteur VIII:C, à l'activité coagulante

Non-Patent Citations (2)

* Cited by examiner, † Cited by third party
Title
EATON ET AL: "PROTEOLYTIC PROCESSING OF HUMAN FACTOR VIII. CORRELATION OF SPECIFIC CLEAVAGES BY THROMBIN,FACTOR XA,AND ACTIVATED PROTEIN C WITH ACTIVATION AND INACTIVATION OF FACTOR VIII COAGULANT ACTIVITY", BIOCHEMISTRY, vol. 25, 1986, pages 505 - 512, XP002019694 *
PITTMAN ET AL: "PROTEOLYTIC REQUIREMENTS FOR THROMBIN ACTIVATION OF ANTI-HEMOPHILIC FACTOR (FACTOR VIII)", PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES,USA, vol. 85, 1988, pages 2429 - 2433, XP002019693 *

Cited By (4)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US6838437B2 (en) 1996-04-24 2005-01-04 University Of Michigan Inactivation resistant factor VIII
US7459534B2 (en) 1996-04-24 2008-12-02 The Regents Of The University Of Michigan Inactivation resistant factor VIII
US8183344B2 (en) 1996-04-24 2012-05-22 University Of Michigan Inactivation resistant factor VIII
EP2206785A1 (fr) 1998-12-31 2010-07-14 Novartis Vaccines and Diagnostics, Inc. Expression améliorée de polypeptides HIV et production de particules de type virus

Also Published As

Publication number Publication date
AU6456096A (en) 1997-02-10

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