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Fusion protein for treating diabetes, and its preparing method and use

A fusion protein, diabetes technology, applied in gene therapy, chemical instruments and methods, hybrid peptides, etc., can solve the problems of dangerous sensitization of polypeptide drugs and poor efficacy of fusion proteins.

Inactive Publication Date: 2007-03-28
王庆华 +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, the results of in vitro experiments show that the fusion protein is not as effective as Ex-4
And, more importantly, peptide drugs can produce dangerous sensitization reactions

Method used

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  • Fusion protein for treating diabetes, and its preparing method and use
  • Fusion protein for treating diabetes, and its preparing method and use
  • Fusion protein for treating diabetes, and its preparing method and use

Examples

Experimental program
Comparison scheme
Effect test

Embodiment Construction

[0111] 1. Plasmid construction

[0112] To construct GLP-1 and mouse IgG-Fc cDNAs sequences, GLP-1 / PCR2.1 and IgG plasmids were used as templates, and the following specific sequences 5'-ccgGATATCgccaccATGGAGACAGACACACTCCTGCTATGGGTACTGCTGCTCTGGGTTCCAGGTTCCACTGGTGACca-3' and 5'-TgctgaagggACctttaccagtg-3' were used as primers The first round of overlapping PCR (overlapPCR). The second round of overlapping PCR was performed using the PCR product as a template to generate the GLP-1-IgG-FccDNA sequence. The amplified product was subcloned into the Bam HI and Eco RV sites of plasmid Vrnew. To construct GLP-1 and human IgG-Fc cDNAs sequences, use GLP-1 / PCR2.1 and IgG plasmids as templates to construct human IgG with the following specific sequences 2 FC (hinge-ch2-ch3), the primers used were 5'-AAGGATATCGATCGCAAATGTTGTGTCGAGTGCCCA-3' and 5'-CGTAAGCTTCATTTACCCGGAGACAGGGAGAG-3'. The carrier used is the same as above. To construct an IgG-Fc control plasmid, PCR was performed with t...

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Abstract

The invention relates to fusion protein used to prevent and cure I type and II type diabetes and its preparation method and application. It supplies the fusion protein which is formed by glucagon analogy peptide GLP-1, GLP-1 mutant and IgG-Fc, its analogy factor lizard salivary gland polypeptide extendin-4 and IgG-Fc, and the application of the above fusion protein and its DNA used in diabetes prevention and therapy. The fusion protein can increase not only GLP-1 effect, but also the affinity and immunological tolerance of ligand, which is excreted with the form of homogeneity dimmer to improve polypeptide drug effect, overcomes the defect of the GLP-1 for short half-life, and simplifies purification process.

Description

technical field [0001] The present invention relates to the field of biotechnology. Specifically, the present invention relates to a fusion protein for preventing and treating type I and type II diabetes, and its preparation method and application. Background technique [0002] Diabetes is one of the main causes of human death and seriously endangers human health. Type I diabetes and type II diabetes are the two main forms of diabetes. Apoptosis, or programmed death, is the main cause of islet cell death in type I and type II diabetes. Both type I and type II diabetics exhibit progressive islet B-cell mass loss and decreased B-cell function. [0003] Insulin therapy is the mainstay of treatment for type 1 diabetes. Pancreatic islet transplantation is also an effective treatment method, but it is restricted due to limited donors, and the recipient (patient) has to supplement immunosuppressant for life. Conventional treatment of type 2 diabetes, including diet, physical e...

Claims

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Application Information

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IPC IPC(8): C07K19/00C12N15/62C12N15/63A61K48/00A61K31/7088A61P3/10
Inventor 王庆华黄晓航
Owner 王庆华
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