Solid-phase synthesis method of human relaxin-2

A technology of solid-phase synthesis and relaxin, which is applied in the field of synthesis of human relaxin-2, can solve the problems of complex B chain synthesis process, unfavorable industrial production, and low yield

Inactive Publication Date: 2011-09-14
山东长肽医药科技有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

This method can improve the yield of relaxin-2, but the yield is only 48%, and the synthesis process of the B chain of relaxin-2 is complicated, which is not conducive to industrial production
[0010] So far, the synthesis technology of single-chain small-molecule peptides has been mature, and the peptide synthesizer can be used to realize automatic synthesis. Although the solid-phase synthesis of the A chain and B chain of relaxin-2 has achieved good yields, the The Yield of Relaxin-2 Solid Phase Synthesis by Linking A Chain and B Chain Is Still Low

Method used

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  • Solid-phase synthesis method of human relaxin-2
  • Solid-phase synthesis method of human relaxin-2
  • Solid-phase synthesis method of human relaxin-2

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0043] pGlu-Leu-Tyr(Bzl)-Ser(Bzl)-Ala-Leu-Ala-Asn(Tmob)-Lys(Z)-Cys(Trt)-Cys(Acm)-His(Bum)-Val-Gly-Cys Solid Phase Synthesis of (Trt)-Thr(Bzl)-Lys(Z)-Arg(Pmc)-Ser(Bzl)-Leu-Ala-Arg(Pmc)-Phe-Cys(4-MeBzl)-O-Resin

[0044] (1) Swell 50 mmol Wang resin in dichloromethane, and wash the resin with N-methylpyrrolidone.

[0045] (2) After dissolving 60mmol Fmoc-Cys(4-MeBzl)-OH with an appropriate amount of N,N-dimethylformamide, add the solution to the resin.

[0046] Get 60mmol 1-oxygen-3-bisdimethylaminocarbonylbenzotriazole boron tetrafluoride salt, 180mmol 1-hydroxybenzotriazole and 60mmol N, N-diisopropylethylamine to prepare a solution, then The solution was slowly added dropwise into the mixed system of resin and Fmoc-Cys(4-MeBzl)-OH, and stirred at room temperature for 3 hours to obtain Fmoc-Cys(4-MeBzl)-O-resin.

[0047] (3) The Fmoc-Cys(4-MeBzl)-O-resin was sucked dry, and washed three times with dichloromethane, using 5ml of dichloromethane for each gram of resin in each wa...

Embodiment 2

[0051] The cyclization reaction of embodiment 2 linear A chains

[0052] pGlu-Leu-Tyr(Bzl)-Ser(Bzl)-Ala-Leu-Ala-Asn(Tmob)-Lys(Z)-Cys(Trt)-Cys(Acm)-His(Bum)-Val-Gly- Cys(Trt)-Thr(Bzl)-Lys(Z)-Arg(Pmc)-Ser(Bzl)-Leu-Ala-Arg(Pmc)-Phe-Cys(4-MeBzl)-O-resin with hydrogen bromide In the mixed solution that acetic acid is made into by volume ratio 3: 7, the mixed solution consumption of hydrogen bromide and acetic acid is that every gram of resin uses 4-9ml, reacts 20 minutes and removes the Trt protecting group of Cys (Trt) at room temperature, will After washing the resin with dichloromethane, pGlu-Leu-Tyr(Bzl)-Ser(Bzl)-Ala-Leu-Ala-Asn(Tmob)-Lys(Z)-Cys(Trt)-Cys(Acm )-His(Bum)-Val-Gly-Cys(Trt)-Thr(Bzl)-Lys(Z)-Arg(Pmc)-Ser(Bzl)-Leu-Ala-Arg(Pmc)-Phe-Cys( Add water, acetonitrile and dimethyl sulfoxide to the 4-MeBzl)-O-resin to prepare a mixed solution of 5:4:1 by volume, and stir at room temperature for 4 hours to carry out the cyclization reaction, wherein the mixed solution of hydrog...

Embodiment 3

[0054] Embodiment 3A chain side chain protecting group and the excision of resin

[0055] Wash the peptide-linked resin obtained in Example 2 and add it to the mixed solution prepared by bromotrimethylsilane, thioanisole, methylene chloride and trifluoroacetic acid in a volume ratio of 3:2:4.5:0.5 , standing at a constant temperature of 10°C for 2 hours, then washing and drying the resin linked to the polypeptide, adding trifluoroacetic acid, dichloromethane, N-methylmorpholine and triethylsilane in a volume ratio of 5.5:2.0:1.5:1.0 to prepare In the mixed solution, it was kept at a constant temperature of 0°C for 2 hours. Then add the resin connecting the peptide chains to a mixed solution prepared by volume 70:8:10:5:7 of trifluoroacetic acid, thioanisole, phenol, dimercaptoethanol and water, and let it stand for 2 hours to form the following structural formula: A chain of single rings exists within the chain:

[0056]

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Abstract

The invention discloses a solid-phase synthesis method of human relaxin-2, which comprises the following steps of: 1) solid-phase synthesis of a single-ring chain A; 2) solid-phase synthesis of a chain B; and 3) chemical synthesis of the human relaxin-2. By using the method to produce the human relaxin-2, the yield is high and can be up to 74-82%. The method disclosed by the invention has the advantages of convenient production steps and no impurity production in the production process, and effectively solves the problems that the solid-phase synthesis of the chain B of the human relaxin-2 isdifficult to complete by the Fmoc strategy and the synthesis process is too complicated.

Description

technical field [0001] The invention relates to a synthetic method of human relaxin-2, in particular to a solid-phase synthetic method of human relaxin-2. Background technique [0002] Human relaxin-2, usually abbreviated as RLN2, and the drug name is also named Relaxin, namely relaxin, is a polypeptide hormone found in the human body. Human relaxin-2 consists of two chains, the A chain contains 24 amino acids and has intra-chain disulfide bonds, the B chain contains 29 amino acids, and the A chain and the B chain are connected by two disulfide bonds. Its structural formula is as follows: [0003] [0004] A series of studies in recent years have found that human relaxin-2 has a significant therapeutic effect on cardiovascular diseases such as heart failure, and can expand blood vessels in the systemic circulation, expand renal blood vessels, and increase arterial compliance. U.S. Patent US5166191 has disclosed the method patent of human relaxin for the treatment of card...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07K14/64C07K1/06C07K1/04
CPCY02P20/55
Inventor 厉保秋厉凌子
Owner 山东长肽医药科技有限公司
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