Fusion protein of GLP-1 analogue and preparation method and application thereof

Abstract

The invention provides a class of novel fusion protein of a GLP-1(glucagon like protein-1) analogue and a preparation method and application thereof. The fusion protein is composed of three ingredients: the GLP-1 analogue, connecting peptide and human serum albumin (HSA). The compound prepared by using the method has the advantages that the production cost is very low, the biological activity is higher, and the stability in vitro and vivo is better. The fusion protein can be used for treating diabetes, obesity and irritable bowel syndrome and other diseases which benefit from fasting plasma glucose reduction, stomach and / or bowel movement inhibition and stomach and / or bowel empty or food intake inhibition.

Description

technical field [0001] The present invention relates to a novel GLP-1 analog fusion protein and a method for preparing the fusion protein. The GLP-1 analog fusion protein is used for treating diabetes and various related diseases or disorders. Background technique [0002] Glucagon-like peptide-1 (GLP-1) and its analogs such as Exendin-4 are widely used to study the treatment of type 2 diabetes, because the GLP-1 polypeptide is digested by protease dipeptidyl peptidase Ⅳ (DPP- IV) is rapidly inactivated, and its half-life in plasma is very short, making it difficult to be widely used clinically; while Exendin-4 is insensitive to the enzymatic degradation of DPP-IV, its stability increases, but its molecular weight is low (4187.61 D), short half-life in vivo, so it must be injected twice a day, which hinders clinical use. Many efforts have been made to solve this technical problem, such as slow-release microspheres, PEG modification, fatty acid chain modification, albumin f...

AI Technical Summary

Problems solved by technology

A large increase in clinical dose will bring two problems: 1. Increase the potential risk of immunogenicity

Therefore, a significant increase in the dose of GLP-1 / HSA fusion protein preparations will inevitably lead to an increase in the risk of antibody production; 2. GLP-1 / HSA fusion protein preparations need to be prepared through extremely complex bioengineering techniques, and the cost per unit protein amount High, the substantial increase in dosage will make the price of the drug unaffordable for diabetic patients

[0005] Secondly, since most of the GLP-1 sequence is in a random coil conformation, it is extremely vulnerable to protease attack and degraded, so the additional second GLP-1 makes Albugon more vulnerable to protease attack and unstable

This instability reflects disadvantages in two aspects: 1. When performing recombinant expression of Albugon, whether it is a yeast expression system with low production cost or a mammalian cell expression system with high production cost, the GLP-1 / HSA fusion protein is susceptible to protease degradation, which not only leads to a decrease in expression, but also produces a lot of inhomogeneous enzymatic hydrolysis products, which makes the final product inhomogeneous; 2. After Albugon is injected into the body, it is also susceptible to protease degradation and fails when it circulates in the body

[0006] In addition, limited to the stability of the product, such products currently need to be stored and transported at low temperature, so it is extremely inconvenient for diabetics to travel

Images