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A kind of synthetic peptide containing glycine, l-cysteine, l-phenylalanine and its application

一种神经系统疾病、药物的技术，应用在生物医药领域，能够解决限制临床应用、副反应严重、剂量过大等问题，达到改善氧化应激损伤、副作用小、改善认知功能障碍和痴呆的效果

Active Publication Date: 2019-09-17
WUHAN HONGYUE MEDICAL SCI INC
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0008] Although both glycine and cysteine ​​have obvious brain protective effects, the current treatment plan is to administer glycine (800mg / kg) or cysteine ​​(500mg / kg) alone peripherally, and the dose is too large, resulting in serious side effects , limiting their clinical application

Method used

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  • A kind of synthetic peptide containing glycine, l-cysteine, l-phenylalanine and its application
  • A kind of synthetic peptide containing glycine, l-cysteine, l-phenylalanine and its application
  • A kind of synthetic peptide containing glycine, l-cysteine, l-phenylalanine and its application

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0059] [Example 1] This patent takes glycine-L-cysteine-L-phenylalanine tripeptide (GCF) as an example to illustrate the preparation process of the solid-phase synthetic peptide as follows:

[0060] 1. Synthesis of Complex I Fmoc-L-Phe-resin:

[0061] The tripeptide of this patent uses the method of solid-phase synthesis, weighs 10.7g of 2-chlorotrityl chloride resin (2-Chlorotrityl Chloride Resin) with a degree of substitution of 0.4mmol / g, puts the resin into a polypeptide reaction tube, and adds Dichloromethane (DCM) 160.5ml, shaken for 30 minutes; filter the DCM solvent through sand core, add 12mmol of Fmoc-L-Phe-OH amino acid, then add 40mmol of N,N-diisopropylethylamine (DIEA ), and finally add a small amount of dimethylformamide (DMF) to dissolve, shake for 1h. Alternately wash 6 times with DMF and DCM. Add 160.5ml of 20% piperidine (piperidine is dissolved in DMF solution), remove the piperidine after 5 minutes, add 160.5ml of 20% piperidine again, remove the piperid...

Embodiment 2

[0078] [Example 2] Cytotoxicity test of the patent small peptide on primary cortical neurons

[0079] In this example, the primary cortical neurons cultured for 10 days were used as the experimental object, the neurons were treated with different concentrations of GCF, and the cell viability was evaluated by cck8, so as to judge the cytotoxicity of GCF, and the phosphorylation level of Akt was used to illustrate Whether GCF has a certain degree of protective effect on nerve cells. It was found that when GCF>=800μM, it has obvious protective effect. In order to determine whether the small peptides of this patent also have a protective effect on the brain, we established an isolated glucose-oxygen deprivation (OGD) model of cortical neurons. Under the condition of OGD injury, the neurons were treated with 500 μM of each small peptide, and evaluated by cck8 The viability of the cells can be used to determine whether each small peptide has a protective effect on neurons. The sma...

Embodiment 3

[0103] [Example 3] Establish a rat cerebral ischemia model, inject fluorescently labeled small peptides intravenously, and observe the ability of small peptides to penetrate the blood-brain barrier as a whole

[0104] 1. The main reagents and instruments of the experiment

[0105]FITC, Wuhan Minghao Biotechnology Co., Ltd.; FITC-GC and FITC-GCF, Wuhan Minghao Biotechnology Co., Ltd.; OCT glue, Beijing Zhongshan Jinqiao Biotechnology Co., Ltd.; pathological tissue slicer, Leitz, Germany, type 1512; Fluorescence microscope, Ningbo Sunny Instrument Co., Ltd.; paraformaldehyde, Sinopharm Chemical Reagent Co., Ltd.

[0106] 2. Experimental principle

[0107] FITC (fluorescein isothiocyanate): It is yellow or orange-yellow crystalline powder with a molecular weight of 389.4, a maximum absorption wavelength of 490-495nm, and a maximum emission wavelength of 520-530nm, showing bright yellow-green fluorescence. The broadest fluorescein.

[0108] FITC-GCF: GCF is fluorescently labele...

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Abstract

The invention belongs to the field of medicines and provides application of a glycine-L-cysteine-L-phenylalanine tripeptide to preparation of medicines for treating stroke.According to animal experiments, by means of intravenous administration, the tripeptide penetrates blood brain barriers to enter brain tissues and cells so as to evidently reduce cerebral infarction volume of rats, and a stroke treatment function is realized.The glycine-L-cysteine-L-phenylalanine tripeptide available for conventional intravenous administration is functionally superior to existing clinical medicines.

Description

[0001] Cross References to Related Applications [0002] This application claims priority from Chinese application 201610080163.4, filed February 5, 2016, which is incorporated herein in its entirety by reference. technical field [0003] The invention belongs to the technical field of biomedicine, and in particular relates to a synthetic peptide and its application in preparing medicine for treating brain injury. Background technique [0004] Stroke is a sudden onset cerebral blood circulation disorder, clinically manifested as symptoms and signs of one-time or permanent brain dysfunction. Stroke is divided into ischemic stroke and hemorrhagic stroke according to its nature. Stroke is a refractory disease that seriously endangers human life and safety. It has the characteristics of high morbidity, high disability and high mortality. In 2008, in the ranking of causes of death announced by the Ministry of Health of my country, stroke ranked No. 1 among cities. Three, the sec...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C07K7/06C07K7/08C07K5/103C07K5/107C07K5/087C07K5/083A61K38/06A61K38/07A61K38/08A61K38/10A61P9/10A61P25/00A61P25/16A61P25/28
Inventor 万芪陈娟庄杨张娅万沙
Owner WUHAN HONGYUE MEDICAL SCI INC
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