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Human pancreas glucagon sample peptide-1-derivative, its production and use

A derivative, GLP-1 technology, applied in the direction of glucagon, hormone peptides, specific peptides, etc., can solve the problems of half-life extension, preparation and purification difficulties, etc., and achieve low production cost, low cost, and simplified purification process Effect

Active Publication Date: 2011-01-12
SHANGHAI INST OF BIOLOGICAL PROD CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Compared with natural GLP-1, these GLP-1 derivatives have significantly prolonged half-life, but they still have defects such as difficult preparation and purification.

Method used

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  • Human pancreas glucagon sample peptide-1-derivative, its production and use
  • Human pancreas glucagon sample peptide-1-derivative, its production and use
  • Human pancreas glucagon sample peptide-1-derivative, its production and use

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0028] According to the amino acid sequence of the GLP-1 derivative, the following fragment with a length of 131bp was synthesized, namely Seq ID No.5, using the preferred codons of Escherichia coli:

[0029] ccagatctgg acgacgacga caagcatgcc gaaggcacct ttaccagcga tgtgagcagc 60

[0030] tatctggaag gccaggccgc caaagaattt attgcctggc tggtgaaagg cagaggcnnn 120

[0031] taaccatgga c 131

[0032] Wherein n118=n120=t, n119=g, this fragment contains enterokinase EK site, GLP-1 derivative gene, stop codon TAA and restriction endonuclease BglII and NcoI site;

[0033] The second step places the coding sequence into an expression vector in a manner suitable for expression of the fusion protein:

[0034] The gene fragment obtained in the first step was double-digested with restriction endonucleases BglII and NcoI, purified and recovered, and the Escherichia coli plasmid pET-32a(+) was double-digested with restriction endonucleases BglII and NcoI, and purified Recover the large fragment, mi...

Embodiment 2

[0045] According to the amino acid sequence of the GLP-1 derivative, the following fragment with a length of 134bp was synthesized, namely Seq ID No.6, using the preferred codons of Escherichia coli:

[0046] ccagatctgg acgacgacga caagcatgcc gaaggcacct ttaccagcga tgtgagcagc 60

[0047] tatctggaag gccaggccgc caaagaattt attgcctggc tggtgaaagg cagaggcnnn 120

[0048] nnntaaccat ggac 134

[0049] Wherein n118=n120=t, n119=n121=n122=g, n123=c, this fragment contains enterokinase EK site, human glucagon-like peptide GLP-1 derivative gene, stop codon TAA and restriction endonuclease BglII and NcoI sites;

[0050] Other steps are the same as in Example 1.

[0051] Example 3 Preparation of the GLP-1 derivative of the present invention by DNA recombination technology: the molecular structural formula of the derivative is GLP-1(7-37)-Xaa38-Xaa39 (Seq ID No.3), wherein Xaa38=Gly, Xaa39=Cys , the operation steps are the same as in Example 2, in the Seq ID No.6 gene fragment synthesized ...

Embodiment 3

[0053] The first step is to synthesize a gene fragment according to the amino acid sequence of the GLP-1 derivative:

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PUM

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Abstract

A human pancreatic hyperglycemic hormone sample peptide-1 derivative, its production and use are disclosed. The molecular formula has 3 structures: GLP-1(7-37)-Xaa38(Seq ID No.2), GLP-1(7-37)-Xaa38-Xaa39(Seq ID No.3) and GLP-1(7-37)-Xaa38-Xaa39-Xaa40(Seq ID No.4); Xaa38, Xaa39 and Xaa40 correspond to any amid-acid of Cys, Ala, Gly, His, Ser and Thr separately. It's simple, cheap and efficient, itadopts DNA recombinant technology and has longer half life period. It can be used to prepare active ingredients of medicine in treatment of diabetes.

Description

technical field [0001] The invention relates to a human glucagon-like peptide-1 derivative (GLP-1 derivative) with human glucagon-like peptide-1 (GLP-1) activity and its preparation and application, belonging to bioengineering technology field. Background technique [0002] Diabetes is a worldwide disease, and the global incidence of diabetes is increasing rapidly. It has become the third chronic disease that seriously threatens human health after tumors and cardiovascular diseases. According to the latest report of the International Diabetes Institute (IDI) in 2003, there are 194 million people with diabetes worldwide. According to the current growth rate, by 2025, the number of patients will reach 333 million. There are more than 40 million diabetic patients in my country. In the early 1980s, the incidence rate of diabetes in my country was only 0.67%. Now the incidence rate in Beijing, Shanghai and other places has exceeded 10%, and this number is still increasing. In th...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C07K14/605C12P21/02C12N15/09A61K38/26A61P3/10
Inventor 黄静吴自荣徐进金明飞吴叶林李静
Owner SHANGHAI INST OF BIOLOGICAL PROD CO LTD
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