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Application of TIMP-4 gene in preparing medicine for treating cardiac vascular disease and repairing scar clinically

A TIMP-4, clinical treatment technology, applied in gene therapy, drug combination, pharmaceutical formulation, etc., to achieve the effects of preventing and treating vascular restenosis, slowing down hyperplasia, and inhibiting hyperplasia

Inactive Publication Date: 2003-02-26
王明生 +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0007] The purpose of the present invention is to overcome the shortcomings and deficiencies of the prior art, in order to alleviate the pain of the patient, improve the success rate of balloon catheter dilatation for coronary heart disease, and prevent the restenosis of blood vessels after PTCA from causing the pain of reoperation for the patient, thereby providing a The application of a TIMP-4 gene in the preparation of drugs for clinical treatment of cardiovascular and scar repair, especially the application of drugs for the preparation of clinical prevention and treatment of restenosis after balloon catheter dilatation for coronary heart disease

Method used

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  • Application of TIMP-4 gene in preparing medicine for treating cardiac vascular disease and repairing scar clinically
  • Application of TIMP-4 gene in preparing medicine for treating cardiac vascular disease and repairing scar clinically
  • Application of TIMP-4 gene in preparing medicine for treating cardiac vascular disease and repairing scar clinically

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Experimental program
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Effect test

Embodiment 1

[0034] (1) Construction of specific expression vectors for mammals

[0035] 1. Obtaining the complete sequence of TIMP-4 gene:

[0036] (1) Transform the amplification vector into Escherichia coli (DH52);

[0037] (2) Select and identify positive bacterial strains in the plate;

[0038] (3) Put the positive strain into LB culture medium, shake and culture at 200rpm at 37°C for about 12 hours;

[0039] (4) extracting the amplification vector from the cultured bacterial strain;

[0040] (5) After cutting the amplification vector with ECOR1 and Xhol endonuclease, separate the DNA fragments in 1.2% Agar gel;

[0041] (6) extract the TIMP-4 gene of complete sequence from the Agar glue of step (5);

[0042] 2. Build PCINTIMP-4

[0043] (1) Cut the pCl-neo vector (purchased from Promega Corporation) with ECOR1 and Xhol endonuclease; (2) Gel purification to obtain the linear pCl-neo vector; (3) T4 Ligase 1:1 molar ratio of the linear pCl-neo vector pCl-neo and the full-length TIMP...

Embodiment 2

[0044] Example 2 Select 20 Wester male rats, weighing about 30 grams, and randomly divide them into experimental and control groups, each with 10 male rats. After intraperitoneal injection of phenobarbital sodium 40 mg / Kg, look up and lie down on the animal operating table, insert a puncture needle into the carotid artery, and use a 2F ball bundle dilation catheter (Arterial Embosection Catheter 2F; Baxter Healthcare Corporation; Vascular systems Division ; irvine, CA92614-5686 USA) 0.2ml water column pressure, damage the carotid artery vascular endothelium, and then remove the catheter to stop the bleeding. In the experimental group, 7 days before the operation, the quadriceps femoris (Thighmuscle) was double measured, using a direct current conductance generator (KROHN-HITE CORP. Model 5700-1 Avon, Massachusetts USA) 50-400HZ, 3.5W, 115 / 230V, Transfect the TIMP-4 gene into mammalian muscle cells, inject TIMP-4 intramuscularly at 100-150V unilaterally, at a dose of 600μg / Kg, ...

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Abstract

In the present invention, the biological TIMP-4 gene medicine contains complete cDNA sequence of TIMP-4 gene and the gene of PCINTIMP-4 vector, and TIMP-4 gene is assembled into specific expressing vector PcI-neo of mammal cell to subclone PCINTIMP-4. TIMP-4 gene protein has the powerful functions of inhibiting MMP enzyme, inhibit cell migration of smooth muscles, reducing the change of smooth muscle cell to become spindle cell and inhibiting the proliferation of connective fiber tissue in blood vessel cavity, so that it can inhibit abnormal repair course in blood vessel cavity for normal endothelium repair. The present invention has important application especially in preparing medicine for preventing blood vessel from becoming narrow again.

Description

technical field [0001] The present invention relates to the application of a kind of T1MP-4 gene biopharmaceutical, particularly relate to a kind of TIMP-4 gene as the application in the preparation clinical treatment of cardiovascular and scar repair medicine, clinical prevention and treatment of coronary heart disease balloon catheter dilatation (PTCA ) application of restenosis drugs. Background technique [0002] As we all know, coronary heart disease is one of the diseases that seriously endanger human health. About 400,000-600,000 people die from this disease in the United States within a year. In China, the incidence rate of coronary heart disease is also increasing year by year. The incidence rate has grown from 10-20 per 10,000 in the 1970s to about 10% in the 1990s. The rapid increase and the large number of patients show the severity of the disease sex. The pathological basis of coronary heart disease is the narrowing of the corona...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61K48/00A61P43/00
Inventor 王明生刘宣力
Owner 王明生
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