Method for rapidly synthesizing semaglutide by supersonic waves

An ultrasonic and fast technology, applied in the field of rapid ultrasonic synthesis of semaglutide, can solve the problems of product yield fragment condensation fragment solubility and condensation difficulty, affect production line synthesis efficiency, and do not consider peptide sequence structure, etc., to achieve reduction Effects of coupling time and removal of Fmoc protecting group, avoiding formation of β-sheet secondary structure, improving purity and synthesis yield

Active Publication Date: 2020-12-25
深圳深创生物药业有限公司
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

The use of these methods for the synthesis of semaglutide basically does not consider the peptide sequence structure and characteristics of semaglutide itself in practical applications, such as the influence of the secondary structure of the peptide on the solid-phase synthesis of the peptide, the protection of the amino acid side chain The influence of groups on solid-phase coupling of polypeptides, the influence of longer side chain aliphatic chain hydrophobicity on solid-phase synthesis of polypeptides, the influence of coupling side reactions on product yield, the solubility and difficulty of condensation of fragment condensation, etc.
[0007] At the same time, as a long peptide with more than 30 amino acids in a main chain and a long side chain modification, the traditional solid-phase synthesis method is used. 3+ hours for a single residue
In terms of production, if you do not work continuously 24 hours a day, the synthesis cycle of a batch of semaglutide often exceeds 10 days, which seriously affects the synthesis efficiency of the production line and wastes a lot of labor costs

Method used

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  • Method for rapidly synthesizing semaglutide by supersonic waves
  • Method for rapidly synthesizing semaglutide by supersonic waves
  • Method for rapidly synthesizing semaglutide by supersonic waves

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preparation example Construction

[0040] The invention discloses a preparation method of semaglutide, and those skilled in the art can learn from the content of this article and appropriately improve the process parameters to realize it. In particular, it should be pointed out that all similar replacements and modifications are obvious to those skilled in the art, and they are all considered to be included in the present invention. The method and application of the present invention have been described through preferred embodiments, and the relevant personnel can obviously make changes or appropriate changes and combinations to the method and application described herein without departing from the content, spirit and scope of the present invention to realize and Apply the technology of the present invention.

[0041] First of all, the present invention designs a reaction device of ultrasonic wave combined with polypeptide solid-phase reaction column, the specific device is as attached Figure 4 As shown, the d...

Embodiment 1

[0048] The preparation of embodiment 1 Fmoc-Gly-Wang resin

[0049] Weigh 10 grams of Wang Resin with a substitution degree of 0.8 mmol / g, add it to the solid-phase reaction column of the ultrasonic polypeptide reaction device, add DMF, and ultrasonically swell for 5 minutes; weigh 3.0 grams (10 mmol) of Fmoc-Gly-OH, HOBt 1.6 gram (12mmol), DMAP 0.1 gram (1mmol), dissolved in DMF, added 2.0ml DIC (12mmol), then added to the reaction column, after 20 minutes of reaction, added 7ml acetic anhydride and 6ml pyridine, mixed and blocked for 30 minutes, washed three times with DCM , the resin was dried after shrinking by methanol, and a total of 12 g of Fmoc-Gly-Wang Resin was obtained, and the detected substitution degree was 0.309 mmol / g.

Embodiment 2

[0050] Example 2 Preparation of Semaglutide Main Chain Peptide Resin

[0051] Take by weighing 3.3 grams (1mmol) of the Fmoc-Gly-Wang resin obtained in Example 1 (degree of substitution is 0.309mmol / g), add in the solid-phase reaction column of the ultrasonic polypeptide reaction device, set the ultrasonic frequency to be 30kHz, and the water bath temperature is Wash with DMF for 3 times at 25°C, and then swell the resin with DMF for 10 minutes. Then, under ultrasonic conditions, the Fmoc protecting group was removed with DBLK for 2 minutes, and then washed 5 times with DMF. Weigh 1.94 g (3 mmol) of Fmoc-Arg(Pbf)-OH, 0.5 g (3.6 mmol) of HOBt, dissolve in DMF, add 0.6 ml DIC (3.6 mmol) in an ice-water bath at 0°C, activate for 5 minutes, and add to the reaction column. The reaction was performed under ultrasonic conditions for 10 minutes, and then the Fmoc protecting group was removed with DBLK for 2 minutes under ultrasonic conditions. Repeat the above operation and couple F...

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Abstract

The invention relates to the field of peptide synthesis, and specifically relates to a method for rapidly synthesizing semaglutide by supersonic waves. The method includes the following steps: (1) under the condition of an ultrasonic water bath, coupling Fmoc on solid phase synthetic resin to protect Gly of alpha-amino; (2) under the condition of the ultrasonic water bath, removing the Fmoc protecting group, and successively performing coupling according to the method of coupling the Fmoc or Boc to protect the amino acid of the alpha-amino and then removing the Fmoc protecting group; (3) underthe condition of the ultrasonic water bath, removing the Alloc protecting group in Lys(Alloc); (4) under the condition of the ultrasonic water bath, coupling Fmoc-AEEA-OH, and then successively coupling Fmoc-AEEA-OH, Fmoc-Glu-OtBu and octadecanedioic acid mono-tert-butyl ester according to the method of removing the Fmoc protecting group and then performing coupling; (5) cracking resin and a sidechain protecting group to obtain crude peptide; and (6) performing purification. The method is simple in process, short in synthesis period, good in effect and high in yield; and therefore, the problems of hard solid phase coupling of residues and low yield of the sequence of the semaglutide due to beta-sheet can be solved.

Description

technical field [0001] The invention relates to the field of polypeptide synthesis, in particular to a method for synthesizing GLP-1 analogue sameglutide, in particular to a method for rapidly synthesizing sameglutide by ultrasonic waves. Background technique [0002] Semeglutide is a long-acting GLP-1 analogue developed by Novo Nordisk. The drug was first approved for marketing as an injection, which only needs to be administered by subcutaneous injection once a week. In 2019, the FDA approved the marketing of oral semaglutide. As the first oral GLP-1 analog diabetes drug, the market prospect of the drug is widely optimistic. [0003] From a structural point of view, semaglutide needs to connect the 20th Lys to two AEEA, γ-glutamic acid and octadecanedioic acid fatty chains, and the 2nd position uses the unnatural amino acid aminoisobutyric acid. Compared with liraglutide, semaglutide has a longer fatty chain and increased hydrophobicity, but semaglutide is modified with s...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07K14/605C07K1/04C07K1/06
CPCC07K14/605Y02P20/55
Inventor 马亚平戴政清张凌云王宇恩付信周迎春王庆磊
Owner 深圳深创生物药业有限公司
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