Polypeptides substances, preparing method and use thereof

A technology of uses and aspects, applied in the preparation methods of peptides, peptide/protein components, chemical instruments and methods, etc., can solve the problems of peptide sequences and uses without relevant patents and literature reports, and achieve low cost, clear structure, and activity. high effect

Active Publication Date: 2008-09-17
大连珍奥药业股份有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0011] However, so far, there are no relevant patents and literature reports for the sequence and use of the polypeptide contained in cardiopeptide

Method used

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  • Polypeptides substances, preparing method and use thereof
  • Polypeptides substances, preparing method and use thereof
  • Polypeptides substances, preparing method and use thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0058] This example relates to the synthesis of a polypeptide (cardiopeptide C).

[0059] The sequence of the polypeptide is Lys-Gly-Ala-Trp-Ser-Asn-Val-Leu-Arg-Gly-Met-Gly-Gly-Ala-Phe

[0060] 1. Synthesis: First, the amino group of the amino acid contained in the polypeptide is protected with an amino protecting group (9-fluorenylidene methyl ester group) (amount of 20-100 g / g), and the wang resin is swollen with chloroethyl ether. The amount of chloroethyl ether used is 50-100 ml / g, and then the first amino acid is added to the resin, thereby immobilizing it on the resin. After washing with dichloromethane, the protecting group of the amino group can be deprotected with hexahydropyridine (10-80%, preferably 20-40%). At the same time, the carboxyl group of the second amino acid was activated by HBTU[2-(1H-benzotriazolyl)-1,1,3,3-tetramethylfurfural-hexafluorophosphate] and di After isopropylethylamine is neutralized, it is added to the resin combined with the first amino a...

Embodiment 2

[0062] This example relates to the chemical synthesis method of cardiopeptide X.

[0063] The amino acid sequence of the cardiac peptide C is: Trp-Ser-Asn-Val-Leu-Arg-Gly-Met-Gly-Gly-Ala-Phe.

[0064] The method used is as in Example 1.

Embodiment 3

[0066] This example relates to the chemical synthesis method of cardiopeptide C.

[0067] The sequence of the polypeptide is Lys-Gly-Ala-Trp-Ser-Asn-Val-Leu-Arg-Gly-Met-Gly-Gly-Ala-Phe.

[0068] Described method comprises the following steps:

[0069] (1) Removal of protecting group Fmoc: take Fmoc-Lys(Boc)-CLTR resin, soak and wash with dichloromethane 3 times, each time for 2min, then drain, add 50% piperidine in dichloromethane solution at room temperature 10mL, reacted for 5min and drained, and then reacted with the same amount of the above solution for another 30min, then drained, and then washed with 8mL-10mL of dichloromethane, ethanol and dichloromethane, drained, and took a small amount of resin (2mg~ 10mg) the total amino group was determined by the salicylaldehyde quantitative free radical method.

[0070] (2) Formation of peptide bonds (DCC-HOBt condensation method): Add the dichloromethane solution of Fmoc-Gly (t-Bu) and the dichloromethane of 1-hydroxybenzotria...

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Abstract

The invention discloses a polypeptide substance, preparation method and application thereof. The invention also discloses extraction process of two polypeptides, and determines molecular weight and amino acid sequence of the polypeptides. The polypeptides are synthesized by the chemical method, and purity analysis and molecular weight measurement results of the polypeptides indicate that molecular ion peak (m/z) completely satisfies theoretical molecular weight. Amino acid analysis result of the polypeptides indicates that amino acid content and the composition of the peptides are in accordance with theoretical values. Amino acid sequence measurement result of the polypeptides indicates primary structure thereof is in accordance with the designed sequence. At the same time, activity measurement result of the polypeptides indicates that the two peptides have activity, and remarkable effect on the treatment of myocardial ischemia, cerebral ischemia, and hepatic ischemia.

Description

technical field [0001] The present invention relates to polypeptide substances, specifically two kinds of active polypeptides, their preparation method and application. Background technique [0002] "Myocardial protection" has been a hot spot in the research of heart surgery and heart surgery in recent years. Recent data show that many changes occur in ischemic and hypoxic cardiomyocytes, including intracellular calcium overload, free radical production, membrane damage, ATP (adenosine triphosphate, ATP) level decline, oxygen depletion and so on. [0003] In order to interfere with myocardial ischemia and protect the myocardium, many drugs have been studied in the past 20 years, such as β-receptor blockers, calcium antagonists, converting enzyme inhibitors, various oxygen free radical scavengers, etc. The clinical effect has not yet been confirmed. None of the existing drugs for the treatment of myocardial ischemia can absolutely reduce myocardial infarction and resist myo...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07K7/08C07K1/20C07K1/06C07K1/04A61K38/10A61P9/10
Inventor 陈玉松李恕王日升梁强曾峥钱小红曹东梁慧敏
Owner 大连珍奥药业股份有限公司
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