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Application of brazilin in preparation of drugs

A Brazilian lignin and compound technology, applied in the field of medicine, can solve the problem of no discovery

Active Publication Date: 2014-12-03
INST OF MATERIA MEDICA AN INST OF THE CHINESE ACAD OF MEDICAL SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0008] Regarding the inhibitory effect of brazilianin on PTKs and the effect on VSMC proliferation and migration, there are no related reports at home and abroad.

Method used

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  • Application of brazilin in preparation of drugs
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  • Application of brazilin in preparation of drugs

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0030] Example 1: Evaluation of the inhibitory effect of brazilnin on PTKs activity.

[0031] In this example, 10 μL of the sample and kinase buffer (50mM HEPES buffer, pH7.4, 20mM MgCl 2 , 0.1 mM MnCl 2 , 1 mM Na 3 VO 4 ) 50 μL of the diluted PTKs crude extract was added to a 96-well microtiter plate coated with the substrate polyglutamic acid-tyrosine (Poly-Glu-Ty, PGT), and incubated at 37°C for 10 minutes. Afterwards, 80 μL of ATP (0.4 mM) diluted in kinase buffer was added, and after incubation at 37°C for 40 min, the reaction solution in the plate was removed, and the microwell plate was washed 5 times with PBS containing 0.1% Tween-20. Add 100 μL / well of horseradish peroxidase-labeled phosphorylated tyrosine-antibody complex and incubate at 37°C for 40 min. Wash the microplate 5 times, add 100 μL / well of TMB chromogenic solution, and react for 10 min at room temperature in the dark. Add 100 μL / well of stop solution, and immediately measure the absorbance value at 4...

Embodiment 2

[0035] Example 2: Effects of brazilianin on PDGF-BB-induced VSMC proliferation.

[0036] In this example, primary cultured VSMCs were used to carry out the anti-PDGF-BB-induced VSMC proliferation experiment of brazilin.

[0037] Detection of the effect of Brazilin on the viability of VSMC cells:

[0038] Take the VSMC (3-6 generations) in the logarithmic growth phase, and use 4×10 4 cells / mL, inoculated into a 96-well plate, and added 100 μL to each well. 37°C, 5% CO 2 After culturing in the incubator for 24 hours, the medium was replaced with serum-free DMEM medium, and the cells were starved for 24 hours to synchronize the cells. Drugs of different concentrations (0, 0.1, 1, 3, 10 μM) were added, and the culture was continued. After 12, 24, and 48 hours respectively, the cells were washed twice with PBS, and the cell viability was detected by the MTT method.

[0039] Detection of the effect of Brazilin on the proliferation of VSMC:

[0040]VSMC cells were treated the sa...

Embodiment 3

[0042] Example 3: Effects of brazilianin on PDGF-BB-induced VSMC migration

[0043] In this example, the in vitro wound healing experiment and the cell transmembrane migration experiment were respectively used to conduct the anti-VSMC migration experiment of brazilianin.

[0044] In vitro wound healing experiments:

[0045] Get the VSMC (3-6 generation) of logarithmic growth phase, with 10 5 cells / mL were inoculated into a 24-well plate, and 400 μL was added to each well, at 37°C, 5% CO 2 After incubating in a humidity incubator for 24 hours, use a 200 μL pipette tip to scratch vertically in the 24-well plate in the shape of a "ten", marking four quadrants A, B, C, and D respectively. After scratching, wash the cells twice with PBS, remove the scratched cells, and take pictures at the position of "cross". That is the T0 period. Afterwards, brazilin at a final concentration of 1, 3, 10 and 30 μM diluted in serum-free medium was added, and 20 ng / mL of PDGF-BB was added after...

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Abstract

The invention discloses application of brazilin in preparation of products for prevention, mitigation and / or treatment of diseases related to protein tyrosine kinase activity abnormal increase and vascular proliferation. Brazilin has the effects of inhibiting protein tyrosine kinase (PTKs) activity and inhibiting proliferation and migration of the rat thoracic aortic vascular smooth muscle cell (VSMC). Brazilin can prevent, mitigate and / or treat PTKs activity abnormal increase related diseases like tumors, restenosis after percutaneous coronary intervention (PCI), atherosclerosis, high blood pressure, multiple sclerosis, diabetic retinopathy, endometriosis, psoriasis and other skin diseases, asthma and other diseases. Brazilin can prevent, mitigate and / or treat vascular proliferation related diseases like restenosis after PCI, atherosclerosis, high blood pressure, neointima formation after vascular transplantation, vascular diseases after organ transplantation, diabetic angiopathies and the like.

Description

technical field [0001] The invention relates to the application of brazilin in the preparation of products for preventing, alleviating and / or treating diseases related to abnormally elevated protein tyrosine kinase activity and diseases related to angiogenesis, and belongs to the technical field of medicine. technical background [0002] Brazil lignin is a compound extracted from the core material of the plant Caesalpinia sappan L.Brazilin, the chemical name is [7,11b dihydrobenz(b)indeno[1,2-d]pyran-3,6a,9,10 (6H)-tetrol], the structural formula is as follows: [0003] [0004] In some Asian countries, it is used as a food or beverage coloring, and it is also used for histological staining. Many studies have also found that brasilin has a variety of biological activities, such as lowering blood sugar, protecting the liver, anti-platelet aggregation, enhancing immune tolerance, anti-inflammation, relaxing blood vessels, and anti-allergic asthma. In recent years, it has ...

Claims

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Application Information

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IPC IPC(8): A61K31/352A61P35/00A61P25/00A61P3/10A61P27/02A61P17/00A61P17/06A61P9/10A61P9/12A61P9/00A23L1/30
Inventor 方莲花杜冠华郭晶武玉洁李莉王守宝
Owner INST OF MATERIA MEDICA AN INST OF THE CHINESE ACAD OF MEDICAL SCI
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