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Long-acting GLP-1 and preparation method and application thereof

A GLP-1, long-acting technology, applied in the field of long-acting GLP-1 and its preparation, can solve the problems of short biological half-life, difficulty in exerting effective therapeutic effect, and poor hypoglycemic effect, so as to improve the hypoglycemic effect and solve the problem Unsatisfactory, the effect of solving many adverse reactions

Inactive Publication Date: 2019-05-03
许娜 +1
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  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0007] In order to solve the problems that glucagon-like peptide-1 (GLP-1) itself has a short biological half-life, poor hypoglycemic effect, and difficulty in exerting an effective therapeutic effect, the present invention provides a long-acting GLP-1 and its preparation method and application

Method used

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  • Long-acting GLP-1 and preparation method and application thereof
  • Long-acting GLP-1 and preparation method and application thereof
  • Long-acting GLP-1 and preparation method and application thereof

Examples

Experimental program
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Effect test

experiment example 1

[0018] Preparation of Experimental Example 1 Long-acting GLP-1

[0019] Preparation of recombinant adenovirus AdV-GLP-1: Using the AdMax packaging system to construct a human pDC316-EGFP / GLP1 gene recombinant adenovirus vector, the resulting shuttle plasmid and helper plasmid pBHGlox(delta)E1 were mediated by liposomes, 293 cells were co-transfected with 3Cre and amplified by HEK293A cells. The PCR method identified that the adenovirus vector containing the GLP-1 gene was successfully constructed. The virus liquid was collected and purified to determine the virus titer. The titer reached 1×10 9 PFU / mL.

[0020] Preparation of human umbilical cord mesenchymal stem cells (hUC-MSCs): thawing P2 frozen hUC-MSCs, centrifuging at 1000r / min for 3min, discarding the supernatant, using Daktronics containing 5% AD (UltraGROTM-Advanced) as the mesenchyme Cells were resuspended in mesenchymal stem cell basal medium (MSCBM) and counted at 1×10 6 piece / cm 2 Density seeding in cell cultur...

Embodiment 1

experiment example 2

[0022] Experimental example 2 Long-acting GLP-1 action time experiment in mice with type 2 diabetes

[0023] 1, Type 2 diabetes mouse model preparation

[0024] After one week of adaptive feeding, C57 / BL6 mice were fed with high-fat and high-sugar Western diet for 4 weeks, fasted without water for 12 hours, and induced type 2 diabetes by a single intraperitoneal injection of STZ (130 mg / kg). , fasting blood glucose was detected with a Roche blood glucose meter; animals with fasting blood glucose ≥ 7.8mmol / ml for 2 consecutive weeks were deemed to have successfully prepared the type 2 diabetes model.

[0025] 2. Animals were randomly divided into groups and drug intervention

[0026] The experimental animals were randomly divided into two groups according to their blood sugar levels, namely the long-acting GLP-1 group and the stem cell control group, with 6 animals in each group. Diabetic model mice in the long-acting GLP-1 group, intramuscularly inject the long-acting GLP-1 ...

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Abstract

The invention relates to long-acting GLP-1 and a preparation method and application thereof, relates to the field of type 2 diabetes mellitus medicines, and solves the problems that GLP-1 is short inself biological half-life, poor in effect of reducing blood sugar, and difficult in exertion of effective treatment effects. The preparation method of the long-acting GLP-1 comprise the step that human umbilical cord mesenchymal stem cells are disinfected with recombinant adenovirus AdV-GLP-1 of 30MOI for 48h or above, so that the long-acting GLP-1 is obtained, and the infection efficiency is 90%or above. The invention relates to an application of the long-acting GLP-1 to preparation of the medicines for treating type 2 diabetes mellitus. The long-acting GLP-1 is used for modifying the humanumbilical cord mesenchymal stem cells, so that the long-acting GLP-1 having high blood sugar reducing activity and long pharmacological action time is obtained. The prepared long-acting GLP-1 is injected to mice suffering from the type 2 diabetes mellitus, and the long-acting GLP-1 achieves long-acting treatment effect in bodies of the mice.

Description

technical field [0001] The invention relates to the technical field of type 2 diabetes drugs, in particular to a long-acting GLP-1 and its preparation method and application. Background technique [0002] Diabetes mellitus is a chronic metabolic disease characterized by insufficient insulin secretion and / or hyperglycemia caused by islet resistance. It has become the third major disease that seriously threatens human health after cardiovascular disease and cancer. Diabetes is mainly divided into type 1 diabetes and type 2 diabetes, of which type 2 diabetes accounts for more than 90%. The prevalence of diabetes in China has reached 9.7%, and the total number of people with diabetes has reached 92.4 million. Diabetes has become one of the most serious health hazards in my country. [0003] At present, the treatment methods for type 2 diabetes mainly include: diet control, proper exercise, oral hypoglycemic drugs, and insulin injection, etc., but none of the above methods can e...

Claims

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Application Information

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IPC IPC(8): C07K14/605C12N7/01C12N15/861A61K38/26A61P3/10
Inventor 许娜向亮刘文森常影杨雪
Owner 许娜
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