Pharmaceutical composition comprising factor vii polypeptides and pai-1 polypeptides

A composition, the technology of PAI-1, applied in the direction of drug combination, surgical drug, pharmaceutical formula, etc., can solve problems such as unsuccessful and difficult to treat

Inactive Publication Date: 2005-03-16
NOVO NORDISK AS
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Once a subject has developed these serious complications, a cascade of events involving many cytokines and

Method used

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  • Pharmaceutical composition comprising factor vii polypeptides and pai-1 polypeptides
  • Pharmaceutical composition comprising factor vii polypeptides and pai-1 polypeptides
  • Pharmaceutical composition comprising factor vii polypeptides and pai-1 polypeptides

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Experimental program
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Effect test

preparation example Construction

[0165] Compound preparation:

[0166] Preferably prepared by recombinant DNA techniques such as those described by Hagen et al. in Proc. Purified human Factor Vila suitable for use in the present invention.

[0167] Also by Broze and Majerus in J.Biol.Chem. 255(4):1242-1247, 1980 and Hedner and Kisiel in J.Clin.linvest., 71 : 1836-1841, Factor VII is produced by the method described in 1983. These methods yield Factor VII without detectable amounts of other coagulation factors. Even further purified Factor VII preparations can be obtained by including additional gel filtration as a final purification step. Factor VII is then converted into activated Factor Vila by known means, for example using several different plasma proteins, such as PAI-11a, IXa or Xa. On the other hand, as described by Bjoern et al. (Research Disclosure (Research Disclosure), 269, September 1986, pp. 564-565), by passing Factor VII through an ion exchange column, such as MonoQ  (Pharmacia Fine Chemi...

Embodiment 1

[0218] Improved hemostatic clot stability by combining factor VIIa and PAI-1

[0219] method:

[0220] Clot lysis test: buffer (20 mM HEPES , 150mM NaCl, 5mM CaCl, pH7.4) Normal human plasma diluted 10-fold was added to a 96-well ELISA plate, and the turbidity at 650nm was measured at room temperature over a period of time. Where indicated, purified human PAI-1 (American Diagnostica, various concentrations) was included.

[0221] Rotational Thromboelastography (roTEG): Each assay was performed with citrated normal human plasma or Factor VIII-deficient plasma (George King Bio-Medical, Inc. #0800) spiked with 5 nM t-PA and analyzed separately Effect of 1 nM FVIIa or the addition of 1 nM FVIIa in combination with 30 nM PAI-1. Coagulation was initiated by the addition of Innovin (2000-fold diluted final concentration, DadeBehring #526945) and calcium (15 mM final concentration) in 20 mM HEPES, 150 mM NaCl, pH 7.4 buffer.

[0222] result:

[0223] Clot lysis test: Addition o...

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Abstract

The present invention relates to a composition comprising factor VII or factor VII related polypeptide and PAI-1 or PAI-1 related polypeptide and its application in treating bleeding attacks.

Description

field of invention [0001] The present invention relates to compositions comprising Factor VII or Factor VII-related polypeptides and PAI-1 or PAI-1-related polypeptides. The present invention also relates to the use of a combination of Factor VII or Factor VII-related polypeptides and PAI-1 or PAI-1-related polypeptides for the preparation of a medicament for treating subjects suffering from bleeding episodes or preventing their bleeding episodes. The invention also relates to methods of treating bleeding episodes in a subject, and to methods of enhancing clot formation in a subject. The invention also relates to kits containing these compounds. Background of the invention [0002] Hemostasis is initiated by the formation of a complex between tissue factor (TF), which is exposed to circulating blood as a result of vessel wall damage, and FVIIa, which is present in circulation in an amount corresponding to about 1% of the total FVII protein amount. This complex anchors to T...

Claims

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Application Information

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IPC IPC(8): A61K38/36A61K38/43A61K38/48A61K38/00A61K38/57A61P1/02A61P1/04A61P7/04A61P9/00A61P13/12A61P17/02A61P19/02A61P41/00A61P43/00
CPCA61K38/4846A61K38/57A61P1/02A61P1/04A61P13/12A61P17/02A61P19/02A61P41/00A61P43/00A61P7/04A61P9/00A61K2300/00A61K38/36
Inventor R·罗伊克耶尔
Owner NOVO NORDISK AS
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