Complete solid-phase synthesis method for liraglutide

Abstract

The invention discloses a complete solid-phase synthesis method for liraglutide. 2-Cl-TrtResin is enabled to serve as a solid-phase carrier. DIC / HOBt is enabled to serve as a condensation agent. After processing of microwave reaction technology, reaction time is shortened, and condensation efficiency is improved. According to side chain modification, novel ivDde side chain protected lysine is adopted and side chain modification synthesis is carried out. During the process, 20% piperidine is adopted to get rid of Fmoc protection until linear chain polypeptide synthesis is finished. Then, after hydrazine hydrate is adopted to get rid of ivDde protection, a side chain modification reaction is carried out. Obtained liraglutide with complete protection on the solid phase carrier is processed by trifluoroacetic acid, and crude liraglutide is obtained. After purification and freeze-drying by a C18 column, pure liraglutide is obtained. After strong negative ion salt conversion and free-drying, acetic acid liraglutide acetate is obtained. The complete solid-phase synthesis method for the liraglutide is simple in operation, short in synthesis cycle, low in production cost, few in accessory substance, high in product yield and beneficial for mass production.

Description

technical field [0001] The present invention relates to a method for synthesizing polypeptides, in particular to a method for all-solid-phase synthesis of liraglutide. Background technique [0002] Liraglutide is a human glucagon-like peptide-1 (GLP-1) analogue produced by yeast through gene recombination technology, which has the functions of lowering blood sugar, reducing body weight, promoting islet cell regeneration and cardiovascular system. Protection and other effects, the clinical application prospects are broad. [0003] The existing production methods and defects of liraglutide are as follows: [0004] 1. Genetic engineering method. Since liraglutide is a non-natural active polypeptide with branched chain modification, the production technology is extremely difficult, the equipment requirements are high, the risk of large-scale production is high, and the cost is high, which is not conducive to industrial production. [0005] 2. The solid-liquid synthesis method ...

AI Technical Summary

Problems solved by technology

[0004] 1. Genetic engineering method. Since liraglutide is a non-natural active polypeptide with branched chain modification, the production technology is extremely difficult, the equipment requirements are high, the risk of large-scale production is high, and the cost is high, which is not conducive to industrial production

[0005] 2. The solid-liquid synthesis method needs to go through several different stages such as fragment solid-phase synthesis, fully protected cutting, fully protected fragment purification, and fragment condensation. The production cycle is long

At the same time, because the fragment is a fully protected polypeptide and is very hydrophobic, it greatly increases the difficulty of purification

This brings the problems of long cycle and high cost to mass production

[0006] 3. The Fmoc-Lys(Alloc)-OH method reported in CN102286092A, because it adopts the liquid-phase synthesis of branched compounds (Palmitoyl-Glu-OtBu), also needs repeated purification

3) The reagent tetraphenylphosphine palladium is expensive, the required reaction time is long (2 hours), and it needs to be carried out under the protection of argon

4) Since the reaction is carried out in a closed state, sampling is difficult, which is not conducive to central control, and brings great risks to large-scale industrial production

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