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New application of coumarin

A technology of coumarin and flavor regulator, applied in the new application field of coumarin, to achieve the effects of increasing insulin content, reducing expression, and high cell proliferation rate

Pending Publication Date: 2022-01-28
广州康臣药业有限公司 +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

At present, there is a lack of effective drugs for diabetic nephropathy in clinic

Method used

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  • New application of coumarin
  • New application of coumarin
  • New application of coumarin

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0070] The MTT experiment involved in this embodiment, the main steps are: take GEnCs cells (glomerular endothelial cells) derived from the kidney in the logarithmic phase, and 5% CO 2 , 37°C for culture, followed by 5.0×10 per well 4 The cells were seeded into a 96-well plate, and a series of concentration gradients of 1.25 μM, 2.5 μM, 5 μM, 10 μM, 20 μM, 40 μM, 80 μM, 160 μM, and 320 μM of coumarin (final concentration after adding to the well) were added, and the control group (Control ) without adding coumarin, at least 4 replicate wells for each concentration, after incubation for 24 hours, add 20 μL MTT solution to each well, continue to cultivate for 4 hours, carefully suck off the culture medium in the wells, add 150 μL dimethyl sulfoxide to each well, place Shake on a shaker for 10 minutes to fully dissolve the crystals. The absorbance of each well was measured at OD490nm in an enzyme-linked immunosorbent assay instrument.

[0071] The results show that coumarin has...

Embodiment 2

[0073] The flow cytometry involved in this embodiment, the main steps include:

[0074] Kidney-derived GEnCs cells were prepared at a density of 3.5×10 5Cells / mL were inoculated in a 6-well plate, and when the cells grew to 75% confluence, they were divided into negative control group (no treatment, pure cells without any treatment, deducting the self-fluorescence of cells, Negative Control), and blank control group (without adding any drugs, Control), model group (H-Glu+Insulin), coumarin group (H-Glu+Insulin+7-Hyd) and comparison drug group (H-Glu+insulin+Irb); wherein, Irb corresponds to irbesartan, the final concentration is 13.5mg / mL, and the final concentration of coumarin is 0.5mg / mL. Take it out of the incubator after 24 hours, discard the culture medium, wash once with sterilized PBS, add ROS staining solution DCFH (10 μM), mitochondrial membrane potential staining solution JC-1 (2.5 μg / mL) or apoptosis staining solution PI / FITC (2:1), and incubate in the dark in th...

Embodiment 3

[0077] The immunofluorescence technique involved in this embodiment is divided into negative control group (without adding any drugs), model group (H-Glu+Insulin), coumarin group (H-Glu+Insulin+7-Hyd) and comparison drugs Group (H-Glu+Insulin+Irb). The final concentration of irbesartan is 13.5mg / mL, and the final concentration of coumarin is 0.5mg / mL.

[0078] The main steps include:

[0079] 1. Preparation of cell suspension: trypsinize GEnCs cells derived from kidney, count and resuspend cells in complete medium.

[0080] 2. Cell slides: Add a small amount of complete medium dropwise to each well of the six-well plate, so that the coverslip and the six-well plate can be bonded together by the tension of the complete medium, and then put the cleaned and sterilized coverslip. The cells were evenly inserted into six-well plates covered with 18mm×18mm coverslips. Put it into the cell incubator, add 2mL complete medium to each well after about 1h-2h when 80% of the cells adher...

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Abstract

The invention relates to a novel application of coumarin, in particular to an application of coumarin in preparation of a medicine for preventing or / and treating diabetic nephropathy, and further, the medicine comprises coumarin and pharmaceutically acceptable auxiliary materials. It is found that coumarin is friendly to kidney-derived GEnCs cells and has an improvement effect on multiple indexes of diabetic nephropathy model rats, and it is indicated that coumarin has a good application prospect in the aspect of preventing and treating diabetic nephropathy.

Description

technical field [0001] The invention relates to the technical field of medicine and pharmacy, in particular to the new application of coumarin. Background technique [0002] Chronic kidney disease (CKD) is chronic kidney structure and dysfunction caused by various reasons (history of kidney damage greater than 3 months), including pathological damage with normal and abnormal renal GFR, abnormal blood or urine components, and imaging studies Abnormal examination, or unexplained decrease in GFR (<60ml / min·1.73m 2 ) more than 3 months, that is CKD. Diseases that cause chronic kidney disease include various primary and secondary glomerulonephritis, renal tubular damage, and renal vascular lesions. According to GFR, chronic kidney disease can be divided into 5 stages. Early detection and early intervention can significantly reduce the complications of CKD patients and significantly improve the survival rate. For the treatment of CKD, including the treatment of the primary di...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61K31/37A61K9/20A61K9/16A61K9/14A61K9/48A61K9/00A61K9/08A61P3/10A61P13/12
CPCA61K31/37A61K9/2059A61K9/1652A61K9/146A61K9/4866A61K9/0019A61K9/0095A61K9/0053A61K9/006A61K9/0043A61P3/10A61P13/12
Inventor 徐友华赵婷婷李政海段婷婷列倍锋李敏仪陈德琪杨军政何宝
Owner 广州康臣药业有限公司
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