Glucagon like peptide-1 mutant polypeptide and preparation method, medicinal composition and use thereof

A glucagon, mutant technology, applied in the direction of glucagon, drug combination, hormone peptide, etc., to achieve the effect of improving compliance attitude

Inactive Publication Date: 2012-02-29
天津拓飞科技有限公司
View PDF16 Cites 27 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] Although the half-life of GLP-1 receptor agonists such as Liraglutide on the market has been extended, pat

Method used

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
View more

Image

Smart Image Click on the blue labels to locate them in the text.
Viewing Examples
Smart Image
  • Glucagon like peptide-1 mutant polypeptide and preparation method, medicinal composition and use thereof
  • Glucagon like peptide-1 mutant polypeptide and preparation method, medicinal composition and use thereof
  • Glucagon like peptide-1 mutant polypeptide and preparation method, medicinal composition and use thereof

Examples

Experimental program
Comparison scheme
Effect test

preparation example Construction

[0052] Another aspect of the present invention also provides a method for preparing the above-mentioned GLP-1 mutant polypeptide, the method comprising: after the GLP-1 mutant polypeptide is prepared in solid phase by a polypeptide synthesizer, adding 50-100 mM DTT (dithiothreitol) or β-Mercaptoethanol is denatured, and then slowly oxidized at 4°C or fed with oxygen to form a disulfide bond.

[0053] Another aspect of the present invention also provides the application of the above-mentioned GLP-1 mutant polypeptide in the preparation of a drug for treating diabetes, and the application of the polypeptide complex in the preparation of a drug for treating and / or preventing obesity.

[0054] Moreover, the pharmaceutical composition is preferably an injection, more preferably a lyophilized powder injection or a solution injection.

[0055] Another aspect of the present invention also provides a pharmaceutical composition comprising the above-mentioned GLP-1 mutant polypeptide.

...

Embodiment 1

[0084] Example 1 In vitro GLP-1 receptor binding experiment of GLP-1 mutant polypeptide

[0085] The receptor binding experiment was carried out using the GLP-1 receptor high expression cell line INS-1 (rat islet cell-1, purchased from the Animal Center of Shanghai Chinese Academy of Sciences). First, INS-1 cells were seeded on a 12-well plate, and the cells were washed with PBS buffer 2 hours before the experiment, and then the cells were fixed with 4% paraformaldehyde for 10 minutes at room temperature. After cell counting, 10 5 Cells / hole were respectively in GLP-1 mutant polypeptide (SEQ ID 2, SEQ ID 3, SEQ ID 4, SEQ ID 5, SEQ ID 6, SEQ ID 7, SEQ ID 8 and SEQ ID9) and natural GLP-1 (SEQ ID ID NO 1) Co-incubate for 2 hours. After washing the unbound polypeptides with PBS, the GLP-1 ELISA kit was used for detection. It can be seen from Table 1 that the GLP-1 mutant polypeptide greatly improves its binding ability to the GLP-1 receptor, compared with GLP-1.

[0086] Table...

Embodiment 2

[0089] Example 2 Experiment of hypoglycemic function of GLP-1 mutant polypeptide in mice

[0090] Give mice intravenous injections of 8 different GLP-1 mutant polypeptides of the present invention (injection dose is 100 μg / kg), with Exendin-4 as positive control, after administration, respectively, 0.5, 4, 6, 8, 12 , Inject glucose (2g / kg) at 24 and 48 hours, measure the blood sugar content half an hour after the injection, from figure 1 It can be seen that Exendin-4 completely loses its hypoglycemic function after 4 hours, but the 8 kinds of GLP-1 mutant polypeptides still have the hypoglycemic function after 48 hours of administration.

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to view more

PUM

No PUM Login to view more

Abstract

The invention discloses a glucagon like peptide(GLP)-1 mutant polypeptide and a preparation method, a medicinal composition and use thereof. The mutant polypeptide is formed by bonding a Cys-containing elongation peptide to the N terminal of the glucagon like peptide-1 mutant, the mutant polypeptide itself folds to form a disulfide bond. The glucagon like peptide-1 mutant polypeptide is used for preparing a medicinal composition for treating diabetes and treating and preventing obesity. For overcoming the drawbacks of short retention time in body and need of daily injection administration of clinic GLP-1 analogue, the invention provides the GLP-1 analogue which has a longer half-life period. With the longer half-life period, the GLP-1 mutant polypeptide is not required to be injected into a patient every day, and can increase the obedience of the patient effectively.

Description

technical field [0001] The present invention relates to the field of diabetes-related medicines, in particular, the present invention relates to a glucagon-like peptide-1 mutant polypeptide with prolonged in vivo half-life of glucagon-like peptide-1 and a preparation method thereof, Pharmaceutical composition and its application. Background technique [0002] Diabetes mellitus is a metabolic disorder syndrome characterized by chronic hyperglycemia associated with genetic factors and multiple environmental factors. Because diabetes is also accompanied by many complications, it has become the third largest health killer after malignant tumors and cardiovascular diseases. In 1984, Glucagon-like peptide-1 (GLP-1) was discovered, which is a 30-amino acid incretin that promotes insulin secretion and biosynthesis, inhibits pancreatic The secretion of glucagon promotes the proliferation of islet cells, inhibits the apoptosis of islet cells, and preserves the sensitivity of cells t...

Claims

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to view more

Application Information

Patent Timeline
no application Login to view more
IPC IPC(8): C07K14/605A61K38/26A61K47/48A61P3/04A61P3/10
Inventor 李瑛
Owner 天津拓飞科技有限公司
Who we serve
  • R&D Engineer
  • R&D Manager
  • IP Professional
Why Eureka
  • Industry Leading Data Capabilities
  • Powerful AI technology
  • Patent DNA Extraction
Social media
Try Eureka
PatSnap group products

Browse by: Latest US Patents, China's latest patents, Technical Efficacy Thesaurus, Application Domain, Technology Topic.

© 2024 PatSnap. All rights reserved.Legal|Privacy policy|Modern Slavery Act Transparency Statement|Sitemap