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Novel fusion polypeptide with anti-cerebral ischemia effect and application thereof

An anti-cerebral ischemia and effective technology, applied in the fields of peptides, drug combinations, cardiovascular system diseases, etc., can solve problems such as adverse reactions and inaccurate curative effects, achieve inhibition of excitotoxic damage, good cross-blood-brain barrier effect, The effect of good application prospects

Inactive Publication Date: 2019-02-01
HEBEI UNIVERSITY OF SCIENCE AND TECHNOLOGY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0003] N-methyl-D-aspartate (NMDA) receptor is an important anti-ischemic target. Researchers have developed a variety of NMDA receptor antagonists, but due to uncertain efficacy or serious adverse reactions disabled

Method used

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  • Novel fusion polypeptide with anti-cerebral ischemia effect and application thereof
  • Novel fusion polypeptide with anti-cerebral ischemia effect and application thereof
  • Novel fusion polypeptide with anti-cerebral ischemia effect and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0022] Example 1. Preparation of polypeptide (YGRKKRRQRRRGESV)

[0023] 1. Peptide synthesis process: first soak the resin (Fmoc-Val-Wang Resin) with DCM for 5-10 minutes, wash with DMF for 5 times, deprotect with 20% hexahydropyridine for 10-20 minutes, and wash with DMF for 5 times. Then add protective amino acid raw materials, condensing agent, etc. to condense for 30 minutes, wash 5 times with DMF, then deprotect—clean—condense—clean, finally deprotect, clean, drain, cut with TFA, and precipitate the cutting fluid with ether. Wash 2-3 times with ether and drain. This is an approximate process of the entire peptide synthesis.

[0024] 2. Peptide synthesis starts from the C-terminus, that is, from right to left. First, the sequence YGRKKRRQRRRGESV is synthesized using conventional amino acid raw materials. The amino acid raw materials used are Fmoc-Tyr(tBu)-OH, Fmoc-Arg( pbf)-OH, Fmoc-Pro-OH, Fmoc-Gly-OH, Fmoc-Gln(Trt)-OH, Fmoc-Asp(otbu)-OH, Fmoc-Lys(Boc)-OH. After condensing t...

Embodiment 2

[0025] Example 2. The effect of polypeptide on the excitotoxicity of glutamate

[0026] The fetal rat hippocampal neuron cells cultured to 13 days in vitro were washed three times with HBSS, and incubated with 50 μM glutamic acid for 20 minutes. The glutamic acid was incubated with distilled water, CV-15 or CA-15, respectively. It is 0.05 and 2 μmol / L. After the incubation, remove all drugs, wash the plate 3 times with HBSS, add maintenance medium, and put it in 37℃, containing 5% CO 2 Continue to culture in the incubator for 24 hours, and use MTT method to detect cell damage.

Embodiment 3

[0027] Example 3. Effect of polypeptides on neurobehavioral scores and mortality of animals after cerebral ischemia

[0028] 1. Establishment and administration of rat MCAO model: Sprague Dawley (SD) rat, male, body weight (300±20) g. The middle cerebral artery occlusion (MCAO) cerebral ischemia model was prepared by the internal carotid artery embolization method. After the rats were anesthetized with 10% chloral hydrate (300 mg·kg-1, ip ), they were fixed on the operating table in the supine position, and the cervical median incision was used to separate the right common carotid artery, external carotid artery, and internal carotid artery , Gently peel off the vagus nerve, ligate all branches of the external carotid artery and internal carotid artery. External carotid artery incision, insert a nylon fishing line with an outer diameter of 0.28 mm (the tip is treated with silicone to make it smooth), turn the common carotid artery bifurcation into the internal carotid artery, an...

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Abstract

The invention relates to a novel polypeptide with anti-cerebral ischemia effect and application thereof. The peptide chain fragment sequence of the polypeptide is YGRKRRQRRRGESV. The polypeptide provided by the invention can enter cells under high speed and high efficiency, significantly inhibits glutamate-induced neuron excitotoxicity injury by interfering the signal transduction of NOS1AP, and effectively resists the cerebral ischemic injury of MCAO model rats. The polypeptide has good application prospects in preparation of drugs treating cerebrovascular diseases.

Description

Technical field [0001] The present invention belongs to the technical field of medicines for cerebrovascular diseases. The present invention relates to a polypeptide with anti-cerebral ischemia effect and its application. Background technique [0002] Cerebral ischemia is a common and frequently-occurring disease with high mortality and disability. So far, the only effective treatment method is thrombolytic therapy, but this therapy is not only inefficient, but it must be administered within a short period of time after cerebral ischemia to be effective. Therefore, it is of great significance to further study the molecular mechanism of cerebral ischemia injury, discover new targets and develop new anti-cerebral ischemia drugs. [0003] The N-methyl-D-aspartate (NMDA) receptor is an important anti-cerebral ischemia target. Researchers have developed a variety of NMDA receptor antagonists, but due to uncertain efficacy or severe adverse reactions Was disabled. In recent years, res...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07K7/08A61K38/10A61P9/10
CPCC07K7/08A61K38/00A61P9/10
Inventor 李孟婷孙勇军高子彬陈玺候丽蓓张福云彭姣陈若莹吕晓倩李晨晨姚军
Owner HEBEI UNIVERSITY OF SCIENCE AND TECHNOLOGY
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