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Amylin-like fusion protein and its encoding gene and application

A fusion protein, amylin technology, applied in the direction of peptide/protein components, hybrid peptides, drug combinations, etc., can solve the problems of affecting the therapeutic effect and short half-life, and achieve the effect of enhancing the curative effect, increasing the concentration, and reducing the product cost.

Active Publication Date: 2019-02-15
CHANGZHOU BIOWIN BIOPHARM
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, its blood half-life is short, about 50 minutes, which affects the therapeutic effect

Method used

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  • Amylin-like fusion protein and its encoding gene and application
  • Amylin-like fusion protein and its encoding gene and application

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Experimental program
Comparison scheme
Effect test

Embodiment 1

[0027] Embodiment 1, Escherichia coli in vitro expression of amylin-like fusion protein

[0028] 1. Synthesis of artificial genes: artificially synthesize the partial or full-length sequences of the genes encoding the amino acid residues of the artificial short peptide and amylin-like peptide in sequence 3 in the sequence table, and add BamH I, 3 A NotI restriction site was added to the 'end to obtain the cloned sequence of sequence 3 in the sequence listing. The synthesized nucleic acid sequence was digested with BamH I and NotI respectively, purified, ligated, and the nucleic acid sequence of the fusion protein was synthesized.

[0029] 2. In vitro expression and purification of amylin-like fusion protein

[0030] (1) Put the above artificially synthesized full sequence into the pGEM-T-EASY plasmid respectively to construct the pGEM-PRAM cloning vector.

[0031] (2) Construction of expression plasmid

[0032] Using the high-efficiency expression plasmid pET21b purchased f...

Embodiment 2

[0036] Embodiment 2, the influence of amylin-like fusion protein of the present invention on rat serum glucose and insulin concentration:

[0037]Twelve male SD rats with a body weight of 235-275 g were taken, and the animals were fasted for 16 h before the experiment. They were randomly divided into two groups. 5 mg / kg amylin-like fusion protein was injected, blood was collected 30 minutes later, and serum glucose and insulin concentrations were measured by routine laboratory testing methods. Results The blood glucose concentration was 3.9±0.9 mol / L, 7.3±1.2 mol / L (p<0.01), and the blood insulin level was 15.0±1.8 mIU / L, 37.9±4.3 mIU / L (p<0.01), indicating that amylin The fusion protein increases serum glucose and insulin concentrations.

[0038]

[0039]

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Abstract

The invention discloses a recombinant amylin-type fusion protein for diabetes treatment and is coding gene and use. The invention provides the amylin-type fusion protein, its coding gene and a method for highly expressing the amylin-type fusion protein. The amylin-type fusion protein is a protein obtained by partial or complete fusion of N terminal amino acid residue of amylin-type peptide and carboxyl terminal (C terminal) amino acid residue sequence of an artificial short peptide. The amylin-type fusion protein can be used as a drug for clinical diabetes treatment, has the characteristics of high efficiency and long action time and can be used as a novel diabetes treatment drug.

Description

technical field [0001] The invention relates to an amylin-like fusion protein, its coding gene and application in the field of bioengineering. Background technique [0002] Diabetes has become one of the major diseases affecting the health of the population, and its prevalence is increasing rapidly with the improvement of living conditions, population aging, and changes in lifestyle. Epidemiological survey showed. There are currently 40 million diabetics in my country, of which 5.6% are type I diabetics and 93.7% are type II diabetics. Other types of diabetes accounted for about 0.7%. Controlling the blood sugar level of patients and reducing complications is still the main treatment. At present, there are many hypoglycemic drugs in clinical use, but patients will develop drug resistance to varying degrees as the medication time prolongs. In 20Q5, the U.S. FDA approved Amylin's Pramlintide (hereinafter referred to as "amylin-like peptide") injection for adults with type ...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C07K19/00C12N15/62A61K38/22A61K47/42A61P3/10
Inventor 刘凤鸣
Owner CHANGZHOU BIOWIN BIOPHARM