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Application of tiliroside in preparation of anti-influenza drug

An anti-influenza drug, the technology of silver satin, applied in the field of chemical biology, can solve the problem that neuraminidase activity has not been reported, and achieve the effect of reducing replication and inhibiting activity

Active Publication Date: 2020-06-12
JINAN UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

At present, the flavonoid compound-silverin, which is isolated and purified from the traditional Chinese medicine snakeberry, has a wide range of biological activities such as anti-inflammation and anti-oxidation according to relevant literature reports, but its activity of inhibiting neuraminidase has not been reported yet.

Method used

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  • Application of tiliroside in preparation of anti-influenza drug
  • Application of tiliroside in preparation of anti-influenza drug
  • Application of tiliroside in preparation of anti-influenza drug

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0035] Determination of Inhibitory Activity of Linlin on Neuraminidase of Clostridium perfringens in Vitro

[0036] (1) Solution preparation

[0037] NA solution: An aliquot of NA freeze-dried powder is stored at -20°C. When in use, take a tube and dissolve it in MES buffer with a concentration of 0.021U / mL. Each dissolved NA is used on the same day.

[0038] Sample to be tested: Dissolve the sample in DMSO.

[0039] MES buffer: containing 32.5mmol / L MES, 4mmol / L CaCl 2 , adjust the pH to 6.5.

[0040] MUNANA: Use MES buffer to dissolve into 0.1mmol / L MUNANA solution.

[0041] (2) Measurement of NA activity using the fluorescent substrate MUNANA

[0042] The NA activity assay was carried out in a 384-well plate, and the sample addition method is shown in Table 2. The specific operation method is: the 384-well plate contains 25 μL diluted active NA solution and 25 μL mixed solution of fluorescent substrate and sample, and the concentration of fluorescent substrate in the mix...

Embodiment 2

[0054] In vitro Inhibitory Activity of Silver Lindenin on Neuraminidase of Oseltamivir-resistant / Sensitive Influenza Viruses

[0055] (1) Source of virus supernatant: A / Puerto Rico / 8 / 34(H1N1)(PR8) and its H274Y mutant strain, from the State Key Laboratory of Respiratory Diseases, Guangzhou Medical University.

[0056] The purified virus was lysed with 1% Triton in PBS buffer to obtain a NAase solution, and the virus protein concentration used was 1 mg / mL.

[0057] (2) Solution preparation

[0058] NA solution: According to the results of the preliminary experiment, select the appropriate concentration of the virus supernatant.

[0059] Sample to be tested: Dissolve the sample in phosphate Tween buffered saline (PBST) containing 0.2% tween20.

[0060] MES buffer: containing 33mmol / L MES, 4mmol / L CaCl 2 , adjust the pH to 3.5.

[0061] MUNANA: Use MES buffer to dissolve into 0.2mmol / L MUNANA solution.

[0062] (3) Measurement of NA activity using the fluorescent substrate M...

Embodiment 3

[0069] Inhibitory effect of silver lintin on the replication of oseltamivir-sensitive and drug-resistant influenza virus strains

[0070] MDCK cells were grown on a 96-well plate and infected with PR8 virus (oseltamivir-sensitive strain or resistant recombinant virus with H274Y mutation) at a multiplicity of infection (MOI) of 0.01 for 2 h (DMEM containing 0.05% bovine serum white protein and 1 μg / mL TPCK), and then the medium containing the virus was removed (wherein the medium refers to the DMEM medium containing 0.05% bovine serum albumin and 1 μg / mL TPCK), and then the medium containing 0, 1.6, 8 , 40, and 200 μM doses of satinin solution in the drug medium were added to the 96-well plate, and after continuing to culture for 24 hours, the virus-infected cells were fixed with 4% paraformaldehyde in PBS solution. Expression of viral proteins in cells was detected by enzyme-linked immunosorbent assay (ELISA) of anti-PR8 mouse serum. Inhibitory activity is shown as the relati...

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Abstract

The invention discloses an application of tiliroside in preparation of an anti-influenza drug. Prevention and treatment effects on influenza viruses are achieved by inhibiting neuraminidase activity.The invention provides a novel neuraminidase inhibitor, namely tiliroside, from a natural product duchesnea indica extract. The tiliroside can inhibit neuraminidase of influenza virus oseltamivir sensitive strains and drug-resistant strains, makes up for defects in the prior art, and is expected to be popularized and used as a novel neuraminidase inhibitor anti-influenza drug.

Description

technical field [0001] The invention relates to the technical field of chemistry and biology, in particular to the application of silver lintin in the preparation of anti-influenza drugs. Background technique [0002] Influenza is a permanent threat to human health worldwide. Influenza virus neuraminidase (NA) is a key enzyme in virus replication, transmission and pathogenesis. Its active site is highly conserved for both influenza A and B viruses, and it is an ideal target for anti-influenza drug research. Currently, there are only four drugs targeting NA on the market, namely zanamivir, oseltamivir, panamivir, and laninamivir. However, with the emergence of drug-resistant strains and drug toxicity problems, it is found New NA inhibitors become an urgent need for drug development today. [0003] Natural product extracts are a rich source of active compounds against this highly infectious pathogen. Discovery of therapeutically active bioactive components from traditional ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61K31/7048A61P31/16
CPCA61K31/7048A61P31/16
Inventor 田海妍江正瑾罗思凡李楚芳盛才敏
Owner JINAN UNIVERSITY
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