A kind of preparation method of polypeptide hm-3

A technology of HM-3 and polypeptide, applied in the field of biochemistry, can solve problems such as troublesome, time-consuming and large loss, and achieve the effect of reducing cost, reducing pollution and high yield

Inactive Publication Date: 2011-12-21
CHINA PHARM UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

In liquid phase synthesis, the product must be separated and purified or crystallized after each peptide grafting to remove unreacted raw materials and by-products. This step is quite time-consuming and cumbersome, and the loss is also great.

Method used

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  • A kind of preparation method of polypeptide hm-3

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0035] 1. Synthesis:

[0036] Weigh 14.7g of Fmoc-Asp(Otbu)-Wang resin, pour it into a 1L glass sand core reaction column, add CH 2 Cl 2 147ml to fully expand the resin.

[0037] Uncapping: Add 25ml of hexahydropyridine / DMF decapping solution, seal it and place it in a shaker for 5 minutes, the temperature is controlled at room temperature, drain the decapping solution after 5 minutes, wash with DMF once in the middle, and then add 20% 25ml of capping solution reacted for 15 minutes

[0038] Washing: drain the capping liquid, wash the resin with DMF for 6 times, drain, take 20 resins in a small test tube, add color detector, and heat at 115°C for 3 minutes.

[0039] Condensation: Weigh 2.025g of protected amino acid and HOBt, dissolve in 15ml of DMF and 2.33ml of DIC, then pour it into a reaction kettle for about 1.5 hours, and control the temperature at about 34°C.

[0040] Washing: Drain the reaction solution, wash the resin with DMF for 3 times, drain it, take 10-20 res...

Embodiment 2

[0062] 1. Synthesis:

[0063] Weigh 16.7g of Fmoc-Asp(Otbu)-CTC resin, pour it into a 1L reaction column with glass sand core, add CH 2 Cl 2 167ml to fully expand the resin.

[0064] Uncapping: Add 25ml of hexahydropyridine / DMF decapping solution, seal it and place it in a shaker for 5 minutes, the temperature is controlled at room temperature, drain the decapping solution after 5 minutes, wash with DMF once in the middle, and then add 20% 25ml of capping solution reacted for 15 minutes

[0065] Washing: drain the capping liquid, wash the resin with DMF for 6 times, drain, take 20 resins in a small test tube, add color detector, and heat at 115°C for 3 minutes.

[0066] Condensation: Weigh 4.815g of protected amino acid and HBTU, dissolve in 15ml of DMF and 2.47ml of DIEA, then pour it into a reaction kettle for about 1.5 hours, and control the temperature at about 34°C.

[0067] Washing: Drain the reaction solution, wash the resin with DMF for 3 times, drain it, take 10-2...

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Abstract

The invention discloses a method for preparing HM-3 polypeptide, which belongs to the field of polypeptide technology in the field of biochemistry, and uses Fmoc-Asp(Otbu)-wang resin or Fmoc-Asp(Otbu)-CTC resin as a starting material , and then use protected amino acids to inoculate dipeptide to octadecapeptide sequentially, fully wash after the peptide incorporation work is completed, then cut the peptide, and post-process to obtain the crude product of HM-3. The crude product was dissolved, purified twice by preparative high performance liquid phase, and finally concentrated and freeze-dried to obtain the pure product. The method not only ensures the synthesis efficiency, but also improves the purity.

Description

Technical field: [0001] The present invention relates to the synthesis and purification method of polypeptide in the field of biochemistry, in particular to the synthesis and purification process of polypeptide HM-3. Background technique: [0002] Due to their non-selective cytotoxicity, traditional tumor treatment chemotherapy drugs have relatively large toxic and side effects on patients and are prone to drug resistance. The current method of inhibiting tumor angiogenesis to inhibit tumor nutrition and metastasis has become a new approach for tumor treatment, and its greatest potential lies in overcoming drug resistance. HM-3 is an angiogenesis inhibitor, which is characterized in that the angiogenesis inhibitor is a polypeptide with integrin and heparin affinity and binding ability, and has good effects on the treatment of gastric cancer, rectal cancer, liver cancer and the like. [0003] Peptide synthesis is a process of repeated addition of amino acids, which can be di...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07K7/08C07K1/20C07K1/06C07K1/04
Inventor 徐寒梅张弛
Owner CHINA PHARM UNIV
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