A novel strategy for production of liraglutide precursor peptide and development of a new long-acting incretin mimic

Abstract

Nowadays, a small number of incretin mimics are used to treat type 2 diabetes mellitus (T2DM) due to their longer half-life. The present study aimed to introduce a novel method for producing the liraglutide precursor peptide (LPP) and developing a potentially new incretin mimic. Here, human αB-crystallin (αB-Cry) was ligated to the LPP at the gene level, and the gene construct was expressed in Escherichia coli with a relatively good efficiency. The hybrid protein (αB-lir) was then purified by a precipitation method followed by anion exchange chromatography. After that, the peptide was released from the carrier protein by a chemical cleavage method yielding about 70%. The LPP was then purified by gel filtration chromatography, and HPLC estimated its purity to be about 98%. Also, the molecular mass of the purified peptide was finally confirmed by mass spectroscopy analysis. Assessment of the secondary structures suggested a dominant α-helical structure for the LPP and a β-sheet rich structure for the hybrid protein. The subcutaneous injection of the LPP and the αB-lir hybrid protein significantly reduced the blood sugar levels in healthy and diabetic mice and stimulated insulin secretion. Also, the hybrid protein exerts its bioactivities more effectively than the LPP over a relatively longer period of time. The results of this study suggested a novel method for the easy and cost-effective production of the LPP and introduced a new long-acting incretin mimic that can be potentially used for the treatment of T2DM patients.

Developmental Trends of Peptide Drugs and Their Quality Assessment using Secondary Structure Analysis

“…Cyclic peptide glycoprotein 2b/ 3a inhibitor X-ray, [47] MS, [48] FT-IR [48] Teriparatide 4117.77 (34 aa) SVSEIQLMHNLGKHLNSMERVEWLRKKLQDVHNF-NH 2 two helices peptide PTH receptor CD, [49] NMR, [50] MS, [51] FT-IR [52] Enfuvirtide 4491.95 (36 aa) Ac-YTSLIHSLIEESQNQQEKNEQELLELDKWASLWNWF-NH 2 Helical peptide HIV fusion inhibitor CD, [53] X-Ray, [53] NMR [54] Ziconotide 2639.14 (25 aa) CKGKGAKCSRLMYDCCTGSCRSGKC-NH 2 (Cyclization at Cys1-Cys16, Cys8-Cys20, and Cys15-Cys25) Cyclic peptide N-type Ca channel blocker NMR, [55] Cryo [56] Insulin detemir 5916.89 (49 aa) CD, [58] X-ray, [59] NMR, [58] FT-IR [60] Liraglutide 3751.2 (32 aa) HAEGTFTSSYLEGQAAKEFIAWLVRGRG-NH 2 (Side chain at Lys20 is modified with palmitoyl glutamic acid) Helical peptide GLP-1 agonist CD, [61] NMR (pdb: 4APD), MS, [62] raman [63] Linaclotide 1526.73 ( ChemistrySelect data of the random coil model peptide, thus the reliability of the secondary chemical shift is dependent on the random coil models. The changes of the secondary chemical shift could indicate the contents of the secondary structure in the peptides.…”

Targeting Protein–Protein Interfaces with Peptides: The Contribution of Chemical Combinatorial Peptide Library Approaches

“…Most of the peptides currently approved for therapy work as agonists and are synthetic copies of naturally occurring molecules or analogs used in replacement therapy. For example, Liraglutide, commercially known as Victoza ® , ranking 19th in drug sales (USD 4.39 billion retail), is a synthetic analog of human glucagon-like peptide 1 (GLP-1) used as a drug to manage type II diabetes (T2DM) but with a longer half-life compared to GLP-1 [93][94][95][96]. Other examples include Oxytocin [97], and Parathyroid hormones [98] and their related peptides used to induce or increase labor and for the treatment of osteoporosis, respectively [99].…”

Russian development for drug independence in endocrinology: comparative analysis of bioequivalence, safety and tolerability of the first domestic liraglutide

“…В настоящее время экспрессия чужеродных генов может осуществляться с использованием не только эукариотических систем, но и прокариотических [35]. Escherichia coli (E. coli), является одной из предпочтительных систем экспрессии (US10851146B2 [31], CN114807205A [36], US2020024321A1 [37]), так как ее генетический фон и механизм регуляции хорошо изучены [35], она способна к размножению и не требует дорогостоящего оборудования [35,38,39].…”