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Prothrombic complex composition

A technology of prothrombin and complex, which can be used in drug combination, coagulation/fibrinolytic factor, enzyme, etc., can solve the problem of not being suitable for large-scale preparation of PPSB concentrate

Inactive Publication Date: 2012-05-16
法国分馏学和生物学实验室
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Therefore, the method of document US-P-4272523 proved unsuitable for large-scale preparation of PPSB concentrates intended for therapeutic use

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0070] Example 1: Experimental conditions carried out for the purification of concentrates of vitamin K-dependent proteins.

[0071] A-Preparation of plasma cryoprecipitate supernatant

[0072] As starting material, plasma cryoprecipitate supernatant obtained by freeze-thawing at 0-3°C and centrifugation of frozen fresh plasma was used.

[0073] To separate the insoluble cryoprecipitate consisting mainly of factor VIII, fibronectin and fibrinogen at temperatures below 4°C, cryoprecipitation was performed upstream of plasma fractionation at temperatures below 2°C.

[0074] The cryopreserve was separated from the supernatant by continuous centrifugation at a temperature close to +4°C. The centrifuged supernatant is called cryosupernatant.

[0075] The cold supernatant essentially contains albumin, immunoglobulins, and other vitamin K-dependent factors including prothrombin (factor II), factor VII, factor IX, factor X, protein C, protein S, and protein Z clotting factor.

[0...

Embodiment 2

[0110] Embodiment 2: experimental result

[0111] A: Study of column capacity

[0112] The capacity of the hydroxyapatite columns was tested with doses of 3, 5, 7 and 9 mL of virus-inactivated PPSB-PI-1 per mL of gel under the same experimental conditions as those described above. No pre-elution was performed.

[0113] The yield calculated for each factor corresponds to the ratio of the total amount of solidification units in the eluate from hydroxyapatite to the total amount of solidification units in PPSB-PI-1. The yields obtained by loading are detailed in Table I below:

[0114] Table I - Factors II, IX, VII and X elution yields depending on loading on the gel

[0115]

[0116] These data are shown in the figure 1 middle. A clear reduction in FX binding to loading was observed.

[0117] The percentage of factor unbound calculated for each factor corresponds to the ratio of the total amount of clotting units in the unbound fraction to the total amount of clotting...

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Abstract

The present invention relates to a method for preparing a composition or a concentrate of a prothrombic complex that includes the II, VII, IX and X coagulation factors, wherein said method includes the steps of providing a supernatant of a plasma cryoprecipitate, applying said supernatant on an anion-exchange resin in order to produce an eluate containing said complex and proteins having a high molecular weight, and applying said eluate on a hydroxyapatite column in order to produce a second eluate containing said complex. The invention also relates to a composition that can be produced by said method.

Description

technical field [0001] The present invention relates to a process for the preparation of a composition or concentrate comprising the prothrombin complex of coagulation factors II, VII, IX and X. The invention also provides compositions obtainable by this method. Background technique [0002] The preparation of concentrates of vitamin K-dependent proteins, more particularly the prothrombin complex (also known as PPSB) containing factors II, VII, IX, and X and / or basis for the prevention or treatment of hemorrhagic accidents in hemophiliacs with deficiencies of certain high molecular weight proteins. [0003] During the bleeding phase, patients undergo PPSB therapy and thus receive large doses of plasma proteins, often co-purified with PPSB, which can lead to the development of secondary effects such as anaphylactic shock, inflammatory-type reactions and tolerance problems. This is especially the case for immunoglobulin M and factors C3a, C4a or C5a (also known as anaphylato...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N9/64C12N9/74
CPCC12Y304/21021C12N9/6437C12Y304/21006C12N9/6432C12Y304/21069C12N9/644C12Y304/21022C12N9/6464C12N9/6429C12N9/647C12Y304/21005A61P7/04C07K1/14C12Q1/56C07K14/745C12N7/04
Inventor 雅克·沙巴
Owner 法国分馏学和生物学实验室
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