Liquid-phase synthesizing method for polypeptide

A liquid-phase synthesis and condensing agent technology, which is applied in the production of peptides and bulk chemicals, can solve problems such as undiscovered pentapeptide synthesis methods, and achieve the effects of easy control of the reaction process, simple production process, and short production cycle

Inactive Publication Date: 2013-12-04
SHANGHAI BAJIAYI PHARMA TECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Monocyte migration inhibitory factor (MLIF) is a pentapeptide extracted from the secretion of Entamoeba histolytica, and its amino acid sequence is Met-Gln-Cys-Asn-Ser, but within the scope understood by the inventors, and No synthetic method of this pentapeptide has been reported

Method used

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  • Liquid-phase synthesizing method for polypeptide
  • Liquid-phase synthesizing method for polypeptide

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0058] a..Asn(Cbz)--Ser(tBu)--OtBu dipeptide preparation process:

[0059] Dissolve tBu-tert-butyl serine (63.5 g, 0.29 mol) and N-benzyloxycarbonyl (Cbz)-L-asparagine (78.0 g, 0.29 mol) in 1200 mL of organic solvent N,N-dimethylformamide In DMF, add condensing agent HOBT (43.6g, 0.32mol), DIC (40.6g, 0.32mol) and 122mL organic amine triethylamine, react at room temperature for 7 hours. After the reaction, the reaction solution was poured into 500 ml of saturated sodium bicarbonate solution and stirred for 10 minutes. After suction filtration, washing with water and drying, Asn(Cbz)-Ser(tBu)-OtBu (128 g) was obtained with a yield of 93.9%.

[0060] The NMR spectrum of Asn(Cbz)-Ser(tBu)-OtBu obtained is:

[0061] 1 H-NMR: (CDCl 3 ,300MHz,J:Hz)δδ7.41(d,1H,J=7.8),7.36-7.29(m,5H),6.49(d,1H,J=8.1),6.24(s,1H),5.76(s ,1H),5.60(m,1H),4.52-4.47(m,1H),3.75(dd,1H,J=3.0,8.7),3.50(dd,1H,J=2.7,8.7),2.88(dd, 1H, J=4.5, 15.6), 2.60 (dd, 1H, J=5.4, 15.6), 1.45 (s, 9H), 1.13 (s, 9H).

[...

Embodiment 2

[0083] a..Asn(Cbz)--Ser(tBu)--OtBu dipeptide preparation process:

[0084] tBu-serine tert-butyl ester hydrochloride (73.5g, 0.29mol) and N-benzyloxycarbonyl (Cbz)-L-asparagine (78.0g, 0.29mol) were dissolved in 1200mL DMF, and the condensing agent HOBT (43.6 g, 0.32mol), DIC (40.6g, 0.32mol) and 122mL triethylamine were reacted at room temperature for 7 hours. After the reaction, the reaction solution was poured into 500 ml of saturated sodium bicarbonate solution and stirred for 10 minutes. After suction filtration, washing with water and drying, Asn(Cbz)-Ser(tBu)-OtBu (130 g) was obtained.

[0085] b.. Remove the Cbz protecting group on the Asn (Cbz)--Ser (tBu)--OtBu dipeptide:

[0086] The above product Asn(Cbz)-Ser(tBu)-OtBu (130 g) was dissolved in 1500 mL of methanol, and 19.5 g of 10% Pd-C (10% by mass of Pd) was added. At room temperature, the reaction was hydrogenated for 2 hours, and the reaction was stopped. After filtering off Pd-C, the solvent was distilled of...

Embodiment 3

[0102] Salt-forming process of Met--Gln--Cys--Asn--Ser pentapeptide:

[0103] Take the pentapeptide Met-Gln-Cys-Asn-Ser (1.0 g), dissolve it in methanol-water (volume ratio, 1:1) solution, add 68.8 mg of sodium hydroxide, stir at room temperature for 2 hours, evaporate methanol under reduced pressure, The sodium salt of the pentapeptide Met-Gln-Cys-Asn-Ser (1.05 g) was obtained by lyophilization.

[0104]The preparation method of the acetic acid and trifluoroacetic acid salt of the pentapeptide Met-Gln-Cys-Asn-Ser is the same as above, that is, the Met-Gln-Cys-Asn-Ser pentapeptide is dissolved in methanol-water (volume ratio, 1:1) Add acetic acid or trifluoroacetic acid to the solution, stir at room temperature for 2 hours, evaporate methanol under reduced pressure, and freeze-dry to obtain acetate or trifluoroacetate of the pentapeptide Met-Gln-Cys-Asn-Ser.

[0105] It is also possible to react the Met-Gln-Cys-Asn-Ser pentapeptide with an acid or a base under the action of a...

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Abstract

The invention discloses a liquid-phase synthesizing method for polypeptide. According to the method, tertiary butyl-serine tert-butyl ester or hydrochloride thereof is taken as an initial raw material; asparaginate, cysteine, glutamine and methionine are connected to the amino groups of the raw material in sequence, so as to obtain products of all steps respectively; the protecting groups on the product Met(Boc)-Gln-Cys(Trt)-Asp-Ser(tBu)-OtBu are removed; purification is carried out to obtain pentapeptide Met-Gln-Cys-Asp-Ser; the pentapeptide is further made into pharmaceutically acceptable salt. The method adopts an artificially synthesizing manner, the raw material and equipment costs are low, the yield is high, and suitability for mass production is realized.

Description

technical field [0001] The invention relates to a method for synthesizing a polypeptide, in particular to a method for synthesizing a polypeptide in liquid phase. Background technique [0002] Ischemic cerebrovascular disease (ischemic stroke) is one of the diseases with the highest morbidity, mortality and disability rate in the world, which greatly threatens the health and quality of life of human beings, especially the elderly, and has become a global Huge burden on social medical resources. At present, there are few effective drugs used in the treatment of ischemic cerebrovascular diseases at home and abroad, and it is of great significance to develop new drugs for the prevention and treatment of ischemic cerebrovascular diseases. [0003] Chinese patent (CN101361961) describes that monocyte migration inhibitory factor (MLIF) and its splice body can significantly reduce the area of ​​brain injury caused by cerebral ischemia-reperfusion in mice, and significantly improve...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07K7/06C07K1/02
CPCY02P20/55
Inventor 张大志蔡瞻张恩德黄景华王习著赵明珠李鹏臧成旭姜志辉
Owner SHANGHAI BAJIAYI PHARMA TECH
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