Preparation of specific anticoagulation matter and its use

An anticoagulation and thrombin technology, which can be applied to medical preparations containing active ingredients, using carriers to introduce foreign genetic material, specific peptides, etc., and can solve problems such as no anticoagulant activity.

Inactive Publication Date: 2012-01-04
INST OF RADIATION MEDICINE ACAD OF MILITARY MEDICAL SCI OF THE PLA
View PDF0 Cites 2 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Of course, the above-mentioned substances have no anticoagulant activity on the part of the body where there is no thrombus

Method used

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
View more

Image

Smart Image Click on the blue labels to locate them in the text.
Viewing Examples
Smart Image
  • Preparation of specific anticoagulation matter and its use
  • Preparation of specific anticoagulation matter and its use
  • Preparation of specific anticoagulation matter and its use

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1I

[0019] Embodiment 1 Preparation of IEGR-HV2 (FH) and LGPR-HV2 (TH) and their function of preventing and treating thrombosis

[0020] 1. Acquisition of FH and TH proteins

[0021] By PCR, the restriction site Xho I and the base sequence corresponding to the coagulation factor Xa recognition sequence IEGR or the thrombin recognition sequence LGPR are introduced into the upstream of the HV2 gene, and the EcoR I restriction site is introduced downstream of the HV2 gene, and the gene is Connected to the pPI C9 plasmid cut with the same restriction enzymes to obtain recombinant plasmids pPIC9-FH and pPIC9-TH. pPIC9-FH, pPIC9-TH, and pPIC9K were digested with BamH I and Sal I and ligated to obtain pPIC9K-FH and pPIC9K-TH. The two recombinant plasmids were electroporated and recombined into the yeast genome, and the expression was induced by methanol. The proteins FH and TH were obtained by separation and purification.

[0022] 2. Analysis of antithrombotic effect and bleeding side...

Embodiment 2

[0057] Preparation of Example 2HSA-EFLGPR-HV2 fusion protein (ATH) and HSA-EFIEGR-HV2 fusion protein (AFH) and its antithrombotic and anticoagulant activity

[0058] Add Xho I and EcoR I restriction sites to the upstream and downstream of the human albumin HSA gene, respectively, to construct the HSA gene without a stop codon into the pPIC9 vector, and use BamH I and EcoR I to double-digest the above vector and the pPIC9K vector. HSA is connected to the pPIC9K carrier; the encoding base of EcoR I restriction site and thrombin / coagulation factor Xa recognition sequence is added at the upstream of hirudin gene by PCR method, and the Not I restriction site is introduced at the downstream of hirudin; EcoR I and Not I double-digest the hirudin gene with thrombin / coagulation factor Xa recognition sequence and connect it to the downstream of HSA of the pPIC9K vector with HSA gene constructed above to form the fusion gene ATH / AFH to obtain the plasmid pPIC9K-ATH / AFH. Linearize the p...

Embodiment 3P-I

[0067] Preparation of embodiment 3P-IEGR-HV2 fusion protein (PFH) and its antithrombotic and anticoagulant activity

[0068] Use the PCR method to add the coding base of the EcoR I restriction site and the polypeptide (EFDAEAYVIEGR) containing coagulation factor Xa recognition sequence in the upstream of the hirudin gene, introduce the Not I restriction site in the downstream of the hirudin, and connect EcoR I and Not. I double-digested the pPI C9K plasmid to obtain the pPI C9K-FH recombinant plasmid. The pPIC9K-FH plasmid was linearized, electroporated and recombined into the yeast genome, and methanol-induced expression. The fusion target protein contains three functional regions of oligopeptide P (the amino acid sequence is DAEAYV), blood coagulation factor Xa recognition sequence and hirudin amino acid sequence.

[0069] In vivo antithrombotic activity of the fusion protein (PFH): The in vivo antithrombotic activity of the fusion protein (PFH) was determined using the Car...

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to view more

PUM

No PUM Login to view more

Abstract

Production of specific anticoagulant substance and its use disclosed. The process is carried out by connecting amino acid sequence into anticoagulant substance, which is recognized and cracked by zymoplasm or blood coagulation factor Xa, or connecting anticoagulant substance with amino acid sequence as oligopeptide to obtain final product.

Description

technical field [0001] The invention belongs to the field of biotechnology, and relates to a protein structure of an anticoagulant substance, a preparation method thereof and an application in preventing and treating thrombosis-related diseases. Specifically, the present invention relates to a new substance formed by linking an anticoagulant substance with an amino acid sequence that can be recognized and cleaved by thrombin or coagulation factor Xa, or an anticoagulant substance and other substances that can be recognized and cleaved by thrombin or coagulation factor Xa New substances formed by linking identified and cleaved amino acid sequences, and the medical use of these new substances. Background technique [0002] Cardiovascular disease is the number one killer that threatens human health and life in recent years. Thrombosis is an important cause of many cardiovascular diseases. Anticoagulants are important drugs for preventing and treating thrombosis. At present, th...

Claims

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to view more

Application Information

Patent Timeline
no application Login to view more
Patent Type & Authority Patents(China)
IPC IPC(8): C07K19/00C12N15/63A61K38/57A61P7/02A61P9/10
Inventor 吴祖泽于爱平靳继德牛晋阳苑宾石炳兴董春娜唐仲雄
Owner INST OF RADIATION MEDICINE ACAD OF MILITARY MEDICAL SCI OF THE PLA
Who we serve
  • R&D Engineer
  • R&D Manager
  • IP Professional
Why Eureka
  • Industry Leading Data Capabilities
  • Powerful AI technology
  • Patent DNA Extraction
Social media
Try Eureka
PatSnap group products

Browse by: Latest US Patents, China's latest patents, Technical Efficacy Thesaurus, Application Domain, Technology Topic.

© 2024 PatSnap. All rights reserved.Legal|Privacy policy|Modern Slavery Act Transparency Statement|Sitemap