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Use of Sema3E / PlexinD1 signaling pathway target drug in preparation of drug for inhibiting neointima formation

A signaling pathway and drug technology, applied in the field of medicine, can solve problems such as lack of intervention measures, and achieve the effect of inhibiting the formation of neointima and inhibiting the proliferation and migration of smooth muscle cells

Inactive Publication Date: 2017-12-22
武汉欧瑞康安生物科技有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, the mechanism by which phenotypic transformation occurs has not been fully elucidated, and effective interventions are still lacking

Method used

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  • Use of Sema3E / PlexinD1 signaling pathway target drug in preparation of drug for inhibiting neointima formation
  • Use of Sema3E / PlexinD1 signaling pathway target drug in preparation of drug for inhibiting neointima formation
  • Use of Sema3E / PlexinD1 signaling pathway target drug in preparation of drug for inhibiting neointima formation

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0023] Drugs targeting the Sema3E / PlexinD1 signaling pathway (this example uses Sema3E recombinant protein as an example) inhibit smooth muscle cell proliferation:

[0024] (1) Extraction of primary aortic vascular smooth muscle cells: 5 8-week-old male C57BL / 6 mice were anesthetized by intraperitoneal injection (30-50 mg / kg) of sodium pentobarbital, and the thoracic aorta of the mice was isolated. The thoracic aorta was soaked in type II collagenase for 30 minutes, and then the adventitia was peeled off under a dissecting microscope. The blood vessel was cultured overnight with 10% fetal bovine serum, and the next day, it was digested again with elastase, DNase, and collagenase for 1-1.5 hours , the digested cells were collected and cultured by centrifugation, and the primary aortic vascular smooth muscle cells of passage 3-6 were used for subsequent experiments.

[0025] (2) Inoculate smooth muscle in a 24-well cell culture plate with 10% fetal bovine serum, place at 37°C, 5...

Embodiment 2

[0035] Drugs targeting the Sema3E / PlexinD1 signaling pathway (the Sema3E recombinant protein is used as an example in this example) inhibit the smooth muscle cell cycle:

[0036]According to the method in Example 1, the primary mouse aortic vascular smooth muscle cells were extracted, inoculated in a 6-well cell culture plate with 10% fetal bovine serum, and placed at 37°C, 5% CO 2 Cultivate overnight in the incubator, starve for 24 hours with serum-free DMEM, and then add the following different reagents to stimulate for 24 hours. The grouping details are as follows:

[0037] a. Add nothing (i.e. blank control);

[0038] b. PDGF-BB recombinant protein of 20ng / ml;

[0039] c. 500ng / ml Sema3E recombinant protein + 20ng / ml PDGF-BB recombinant protein;

[0040] d. 500ng / ml Sema3E recombinant protein+20ng / ml PDGF-BB recombinant protein+2.5ug / mlrmPlexinD1. The rmPlexinD1 is a recombinant protein of PlexinD1, which is purchased from R&D with a catalog number of 4160-PD.

[0041]...

Embodiment 3

[0043] Drugs targeting the Sema3E / PlexinD1 signaling pathway (this example uses Sema3E recombinant protein as an example) inhibit smooth muscle cell migration:

[0044] (1) Transwell experiment: According to the method in Example 1, primary mouse aortic vascular smooth muscle cells were extracted, starved overnight in DMEM medium, digested and resuspended, using 8um Transwell chamber, containing 5.0× 10 4 Add 200ul of DMEM to the upper chamber of each cell, add 500ul of serum-free DMEM to the lower chamber, and then add the following different reagents to stimulate, as follows:

[0045] a. Add nothing (i.e. blank control);

[0046] b. PDGF-BB recombinant protein of 20ng / ml;

[0047] c. 1ng / ml Sema3E recombinant protein + 20ng / ml PDGF-BB recombinant protein;

[0048] d. 10ng / ml Sema3E recombinant protein + 20ng / ml PDGF-BB recombinant protein;

[0049] e. 100ng / ml Sema3E recombinant protein + 20ng / ml PDGF-BB recombinant protein;

[0050] f. 500ng / ml Sema3E recombinant prote...

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Abstract

The invention discloses the application of a drug targeting the Sema3E / PlexinD1 signaling pathway in the preparation of a drug for inhibiting neointima formation. The applicant finds that the Sema3E / PlexinD1 signaling pathway can act on smooth muscle cells to inhibit their proliferation and migration. The production of platelet-derived growth factor (PDGF) in endothelial cells, on the other hand, by activating the Sema3E / PlexinD1 downstream signaling pathway, the dual effect significantly inhibits the proliferation and migration of smooth muscle cells and thus inhibits the formation of neointima, which is used for atherosclerosis and stent implantation. Restenosis and other vascular proliferative diseases provide new ideas.

Description

technical field [0001] The invention belongs to the technical field of medicine, and more specifically relates to the application of a drug targeting the Sema3E / PlexinD1 signaling pathway in the preparation of a drug for inhibiting neointima formation. Background technique [0002] Atherosclerosis is an important cause of ischemic heart disease (coronary heart disease) and ischemic stroke. Ischemic cardiovascular and cerebrovascular diseases caused by atherosclerotic lumen stenosis or blockage have become the number one killer of Chinese people's health. . Percutaneous transluminal angioplasty (PTA) uses balloon inflation to dilate occluded or narrowed blood vessels and restore blood supply to normal. Among them, transluminal coronary angioplasty (PTCA) has been widely used in the treatment of ischemic heart disease. Over the decades, PTA techniques have made great strides in success rates and reductions in acute complications. However, about 30% to 50% of the patients' d...

Claims

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Application Information

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IPC IPC(8): A61K45/00A61P43/00A61P9/10
CPCA61K45/00
Inventor 胡波李亚男吴介洪
Owner 武汉欧瑞康安生物科技有限公司
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