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Application of neuraminidase and inhibitor in preparation of medicine for treating myocardial fibrosis and ventricular hypertrophy

A neuraminidase, ventricular hypertrophy technology, applied in the field of biomedicine, can solve the problem of undiscovered neuraminidase correlation and other issues

Active Publication Date: 2020-02-07
CHINA PHARM UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0006] In subsequent studies, the applicant also found that neuraminidase is related to various diseases. After searching, no prior art has been found to disclose the relationship between neuraminidase and these diseases

Method used

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  • Application of neuraminidase and inhibitor in preparation of medicine for treating myocardial fibrosis and ventricular hypertrophy
  • Application of neuraminidase and inhibitor in preparation of medicine for treating myocardial fibrosis and ventricular hypertrophy
  • Application of neuraminidase and inhibitor in preparation of medicine for treating myocardial fibrosis and ventricular hypertrophy

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0020] 1. Experimental method

[0021] After 12-week-old SHR male rats (body weight 200-250g) of SPF grade were fed freely for 3 days, their blood pressure was measured adaptively 5 times. Divided into 5 groups, 10 rats in each group: model group and zanamivir, oseltamivir phosphate, coptisine, salvianolic acid B administration group. Another 12-week-old normotensive WKY male rats were used as the control group. Rats in the administration group were given 0.2mg / kg / d zanamivir (i.v.), 5mg / kg / d oseltamivir phosphate (p.o.), 40mg / kg / d coptisine (p.o.), 40mg / kg / d salvianolic acid B (p.o.), the model group and the control group were intragastrically administered with an equal volume of 0.5% CMC-Na, and the administration continued for 8 weeks.

[0022] After 8 weeks of administration, the following indicators were measured in each group of rats according to conventional methods:

[0023] 1. Myocardial hypertrophy: expressed by left ventricular mass index (LVMI). LVMI is equal ...

Embodiment 2

[0032] 1. Experimental method

[0033] Extraction of primary cardiomyocytes: Isolation of cardiomyocytes and cardiac fibroblasts by differential adherence. Purchase SD suckling rats born 2-3 days old, wipe them with alcohol twice; use ophthalmic scissors to cut the ribs obliquely upwards under the xiphoid process of suckling rats, expose the heart, cut the heart, and put it into pre-cooled PBS solution containing double antibodies , the auricle was carefully removed with ophthalmic curved scissors, and then the heart was cut into uniform tissue pieces of 1mm×1mm×1mm, and washed three times with PBS repeatedly. Transfer to Erlenmeyer flask, add 6-7ml of trypsin (0.08%), digest at a constant speed in a 37°C water bath for 5 minutes, discard the supernatant, and then collect the supernatant every 3 minutes after digestion, stop digestion with culture medium, 1500rpm Centrifuge for ×5min, resuspend the cells in high-glucose DMEM medium containing 10% fetal bovine serum and 1% dou...

Embodiment 3

[0045] The commercially available neuraminidase inhibitor screening kit P0309 (Beyotime, Beyotime) was used to test the inhibitory activity of salvianolic acid B in vitro, and the positive control drug was oseltamivir phosphate. Add 70 μL of buffer solution and 10 μL of neuraminidase solution to each well of a 96-well plate, then add 10 μL of different concentrations of the test solution, shake and mix, incubate at 37°C for 5 minutes, add 10 μL of the solution containing the fluorescent substrate, shake and mix Homogenize, incubate at 37°C for 30min, and perform fluorescence measurement, wherein the excitation wavelength is 322nm and the emission wavelength is 450nm. The inhibition rates of different test solutions were calculated according to the fluorescence readings, and the IC50 values ​​of the positive control drugs oseltamivir phosphate and salvianolic acid B were further obtained. The results are shown in Table 3.

[0046] Table 3 IC50 values ​​of positive control drug...

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Abstract

The invention discloses an application of neuraminidase and inhibitors thereof in preparation of drugs for treating myocardial fibrosis and ventricular hypertrophy. Zanamivir and oseltamivir phosphateare known as effective inhibitors of neuraminidase, and an inhibitory effect of coptisine on neuraminidase also is published in an applicant previous patent application; salvianolic acid B is also found to be able to inhibit the activity of neuraminidase in vitro, and is a neuraminidase inhibitor; experiments prove that the multiple kinds of neuraminidase inhibitors can effectively inhibit the activity of neuraminidase in myocardium in vivo, then inhibit myocardial fibrosis and ventricular hypertrophy, and can be used for treating heart disease caused by myocardial fibrosis and ventricular hypertrophy. Therefore, neuraminidase and the inhibitors thereof can be used for preparation of the drugs for treating myocardial fibrosis and ventricular hypertrophy.

Description

technical field [0001] The invention belongs to the field of biomedicine and relates to the application of enzymes and enzyme inhibitors, in particular to the application of neuraminidase and its inhibitors in the preparation of drugs for treating myocardial fibrosis and ventricular hypertrophy. Background technique [0002] Neuraminic acid is a class of natural sugar and acid compounds widely present in organisms. It is now confirmed that there are more than 50 natural derivatives of neuraminic acid, and N-acetylneuraminic acid and N-glycolylneuraminic acid are the more common ones. Neuraminic acid is usually linked to the end of glycoconjugates such as glycoproteins and glycolipids in the form of short chain residues. Neuraminic acid is an important biological information transfer molecule, and the neuraminidative modification of cell surface glycoproteins and glycolipids plays a vital role in many biological processes, including cell adhesion, antigen recognition and sig...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): A61K38/47A61K45/00A61K31/351A61K31/215A61K31/4375A61K31/343A61P9/04
CPCA61K31/215A61K31/343A61K31/351A61K31/4375A61K38/47A61K45/00C12Y302/01018
Inventor 齐炼文张蕾李萍
Owner CHINA PHARM UNIV
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