Application of salvia miltiorrhiza extracts in preparing protein tyrosine phosphatase 1B inhibitor and drug for preventing and/or treating type 2 diabetes

A technology of tyrosine phosphatase and type 2 diabetes, which is applied in the field of medicine and can solve problems such as insufficient research

Inactive Publication Date: 2019-05-31
ANHUI UNIVERSITY OF TECHNOLOGY AND SCIENCE
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, according to the literature search, there is still basically no relevant literature reporting the relationship betw

Method used

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  • Application of salvia miltiorrhiza extracts in preparing protein tyrosine phosphatase 1B inhibitor and drug for preventing and/or treating type 2 diabetes
  • Application of salvia miltiorrhiza extracts in preparing protein tyrosine phosphatase 1B inhibitor and drug for preventing and/or treating type 2 diabetes
  • Application of salvia miltiorrhiza extracts in preparing protein tyrosine phosphatase 1B inhibitor and drug for preventing and/or treating type 2 diabetes

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0030] Example 1 Evaluation of inhibitory activity of compound salvianolic acid C, salvianolic acid A, isosalvianolic acid C or salvianolic acid A methyl ester on PTP1B

[0031] The PTP1B inhibitory activity of compounds salvianolic acid C, salvianolic acid A, isosalvianolic acid C or salvianolic acid A methyl ester was evaluated by in vitro enzymatic experiments.

[0032] 1.1 Preparation of reagents and standard solutions

[0033] (1) 75mM phosphate buffer (PB, pH 7.4): Weigh KH 2PO 4 0.0956g, K 2 HPO 4 0.6946g, EDTA1.862mg, dissolved with ultrapure water and diluted to 50mL. Freshly prepared before each experiment, used to dissolve diluted samples and other reagents;

[0034] (2) PTP1B solution: Accurately weigh an appropriate amount of PTP1B, dissolve it with 75mM PB solution and prepare a 10U / mL PTP1B working solution, blow the enzyme solution with a pipette, and store it on ice until use;

[0035] (3) Substrate preparation: Accurately weigh an appropriate amount o...

Embodiment 2

[0046] Example 2 Compounds Salvianolic Acid C and Salvianolic Acid A Determination of Inhibition Kinetics of PTP1B

[0047] In vitro enzyme kinetic experiments were used to determine the inhibitory activity of compounds salvianolic acid C and salvianolic acid A on PTP1B.

[0048] 1.1 Preparation of reagents and standard solutions

[0049] (1) 75mM phosphate buffer (PB, pH 7.4): Weigh KH 2 PO 4 0.0956g,K 2 HPO 4 0.6946g, EDTA1.862mg, dissolved with ultrapure water and diluted to 50mL. Freshly prepared before each experiment, used to dissolve diluted samples and other reagents;

[0050] (2) PTP1B solution: Accurately weigh an appropriate amount of PTP1B, dissolve it with 75mM PB solution and prepare a 10U / mL PTP1B working solution, blow the enzyme solution with a pipette, and store it on ice until use;

[0051] (3) Substrate preparation: Accurately weigh an appropriate amount of 4-Nitrophenyl-phosphate (p-NPP), add 75mM PB solution to dissolve, and prepare a substrate wo...

Embodiment 3

[0059] Example 3 Compounds Salvianolic Acid C and Salvianolic Acid A Determination of Inhibitory Reversibility of PTP1B

[0060] 1.1 Experimental steps

[0061] (1) Add 100 μL of the sample solution to be tested at different concentrations on a 96-well plate, then add 50 μL of PTP1B solution at different concentrations (0.1-0.5 U / mL), and use the same volume of PB as the blank control group, and each group is parallel Set up 3 replicate holes. Place the enzyme system on a microplate reader at 37°C and incubate for 10 min, read once at a wavelength of 405 nm, and record the absorbance value.

[0062] (2) Subsequently, 50 μL of substrate p-Nitrophenyl phosphate (p-NPP) (0.5 mM) was added to the enzyme system to start the enzyme reaction system, read at a wavelength of 405 nm every 30 s for a total of 10 min, and record the absorbance value.

[0063] (3) Data processing: use MS Excel 2013 and Graphpad Prism v6.0 to comprehensively analyze and process the experimental data obtai...

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Abstract

The invention discloses application of salvia miltiorrhiza extracts in preparing a protein tyrosine phosphatase 1B inhibitor and a drug for preventing and/or treating type 2 diabetes. The salvia miltiorrhiza extracts salvianolic acid C, salvianolic acid A, danshinolic acid C or salvianolic acid A methyl ester can significantly inhibit the activity of protein tyrosine phosphatase 1B, further increase the glucose uptake rate, reduce the plasma glucose levels and prevent and/or treat the type 2 diabetes. The salvianolic acid C, the salvianolic acid A, the danshinolic acid C or the salvianolic acid A methyl ester are used as active ingredients to make a pharmaceutical preparation which can be used as the protein tyrosine phosphatase 1B inhibitor to prevent and/or treat the type 2 diabetes.

Description

technical field [0001] The invention belongs to the technical field of medicine, and specifically relates to the application of salvia miltiorrhiza extract in the preparation of protein tyrosine phosphatase 1B inhibitors and drugs for preventing and / or treating type 2 diabetes. Background technique [0002] Diabetes is a metabolic disease that seriously threatens human health. Hyperglycemia is its main physiological feature. It can be divided into type 1 diabetes (T1DM) and type 2 diabetes (T2DM). Among them, type 1 diabetes is caused by insulin deficiency, accounting for about 5-10% of the diabetic population, while T2DM is mainly caused by insulin resistance, which is the most common type, accounting for more than 90% of the diabetic population. And often accompanied by a variety of serious complications, such as uremia, renal failure, neuropathy and cardiovascular disease. [0003] In recent years, a variety of effective therapeutic approaches have been used in the clini...

Claims

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Application Information

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IPC IPC(8): A61K36/537A61P3/10
Inventor 唐红进徐晨晨张婧妍张高曼刘亚婷
Owner ANHUI UNIVERSITY OF TECHNOLOGY AND SCIENCE
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