Glucose kinase derivant for inhibiting platelet aggregation and thrombase activity, and its preparing method

A technology of platelet aggregation and derivatives, which is applied in the fields of genetic engineering and biothrombolytic drug preparation, can solve the problems of antigenic influence, thrombolytic blockage, etc., expand the application range, inhibit and block thrombin activity, and inhibit platelet aggregation Effect

Active Publication Date: 2006-10-25
INST OF BASIC MEDICAL SCI ACAD OF MILITARY MEDICAL SCI OF PLA
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Problems solved by technology

Clinical studies have shown that SAK is the only new thrombolytic drug comparable to tPA in terms of thrombolytic effect and specificity. However, because it is a foreign protein, its antige

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  • Glucose kinase derivant for inhibiting platelet aggregation and thrombase activity, and its preparing method
  • Glucose kinase derivant for inhibiting platelet aggregation and thrombase activity, and its preparing method
  • Glucose kinase derivant for inhibiting platelet aggregation and thrombase activity, and its preparing method

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Embodiment Construction

[0035] The invention provides a staphylokinase derivative capable of inhibiting platelet aggregation and thrombin activity and a preparation method. The present invention is described as follows in conjunction with accompanying drawing:

[0036] 1. Experimental materials and equipment

[0037] 1.1 Strains: Staphlococcus aureus (Staphlococcus aureus 1.1476) was purchased from the National Culture Collection Center (CGMCC), E.coli DH5, BL21, etc. (preserved strains of the Institute of Basic Medical Sciences, Academy of Military Medical Sciences).

[0038]1.2 Plasmid: pT-SAK-M (constructed by the Institute of Basic Medicine, Academy of Military Medical Sciences), the pBV220 expression vector is 3.66kb in size, and is a temperature-sensitive high-efficiency expression vector containing P R P L Tandem promoter, cIts857 temperature-sensitive regulatory gene and P R P L The presence of the vector can transform any recipient bacteria, and the SD sequence is followed by multiple cl...

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Abstract

The invention discloses the grape dashing enzyme ramification dealing with the blood platelet assembling and the thrombin active ability and the producing process belonging to the domain of the gene engineering and the producing art of the biology dissolving bolt medicament. The nucleotide acid serial of the grape dashing enzyme ramification restraining the blood platelet assembling and the thrombin active ability is disclosed. The producing process fuses the RGD series, the C end series of the hirudin and the constructed low antigen and high active ability SAK breaking body gene using the gene recombining technology outer the body, at the same time, the Xa gene incising location point and the flexible connecting arm and it's C end amino acid is decorated, the another amino acid is appended. The fusing albumen expressing carrier is constructed to gain the efficient expressed recombining SAK ramification using the double enzyme chipping and the PCR appraising alkalescence of the constructed breaking body. The ramification has the function of restraining the blood platelet assembling and interdicting the thrombin active ability and establishes the base of developing the new generation dissolving bolt medication. So the applied range of the new SAK ramification is expanded largely.

Description

technical field [0001] The invention belongs to the technical scope of genetic engineering and biothrombolytic drug preparation, and particularly relates to a staphylokinase derivative that inhibits platelet aggregation and thrombin activity and a preparation method. Background technique [0002] Cardiovascular and cerebrovascular diseases are currently the number one killer that endangers people's health, and acute myocardial infarction is the main cause of death of patients. There are still many problems to be solved urgently in the thrombolytic effect, specificity, toxic and side effects and drug price of the currently commercially available thrombolytic drugs. [0003] Natural staphylokinase (Staphylokinase, SAK) is a single-chain protein synthesized by lysogenic phage of Staphylococcus aureus, consisting of 136 amino acids, without disulfide bonds, and with a molecular weight of 15.5kDa. In the 1990s, the fibrin selectivity of SAK was discovered, which attracted widesp...

Claims

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Application Information

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IPC IPC(8): C12N15/57A61K38/48A61P7/02C12N9/50
Inventor 徐东刚邹民吉王旻蔡欣王金凤陈光宇刘深徐涛
Owner INST OF BASIC MEDICAL SCI ACAD OF MILITARY MEDICAL SCI OF PLA
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