A two-step method preparation of semaglutide through solid-phase synthesis and inclusion body expression

Dezheng Peng; Yang Li; Linlin Si; Bo Zhu; Peng Wu; Yibang Li; Dongfang Tang; Yu Liu; Yunxiao Zhang

doi:10.1016/j.pep.2024.106477

A two-step method preparation of semaglutide through solid-phase synthesis and inclusion body expression

Protein Expr Purif. 2024 Mar 23:219:106477. doi: 10.1016/j.pep.2024.106477. Online ahead of print.

Authors

Dezheng Peng¹, Yang Li¹, Linlin Si¹, Bo Zhu¹, Peng Wu¹, Yibang Li¹, Dongfang Tang², Yu Liu³, Yunxiao Zhang⁴

Affiliations

¹ School of Chemistry and Chemical Engineering, Hunan Institute of Science and Technology, Yueyang, 414006, Hunan, China.
² Department of Chemical and Biological Engineering, Hunan University of Science and Engineering, Yongzhou, 425199, Hunan, China.
³ School of Chemistry and Chemical Engineering, Hunan Institute of Science and Technology, Yueyang, 414006, Hunan, China; Shenzhen Pengtai Biotechnology Co., Ltd, Shenzhen, 518000, Guangdong, China. Electronic address: hiliu2000@126.com.
⁴ School of Chemistry and Chemical Engineering, Hunan Institute of Science and Technology, Yueyang, 414006, Hunan, China; Key Laboratory of Hunan Province for Advanced Carbon-based Functional Materials, School of Chemistry and Chemical Engineering, Hunan Institute of Science and Technology, Yueyang, 414006, Hunan, China. Electronic address: zhangyx@hnist.edu.cn.

PMID: 38527576
DOI: 10.1016/j.pep.2024.106477

Abstract

Semaglutide is currently the most promising antidiabetic drug, especially for the treatment of type 2 diabetes mellitus, due to its excellent efficacy in glycemic control and weight loss. However, the production of semaglutide remains high cost, and high yield, low cost, and high purity still remains a challenge. Herein, we reported a convenient and high-yield strategy for the preparation of semaglutide through fragmented condensation coupling, involving solid-phase peptide synthesis of tetrapeptide and on-column refolding and on-column enzyme cleavage based inclusion body expression of Lys²⁶Arg³⁴GLP-1 (11-37) with fused protein tags in an X-Y-D4K-G pattern. The optimized N-terminal protein tag significantly boosts inclusion body expression level, while on-column refolding and on-column enzyme cleavage avoid precipitation, enhancing efficiency and yield together with one-step purification. The successful preparation of semaglutide is expected to achieve large-scale industrial production with low cost, high yield and high purity.

Keywords: Fmoc-based peptide synthesis; Inclusion body expression; Lys(26)Arg(34)GLP-1 (11–37); On-column refolding; Semaglutide.