Looking for breakthrough ideas for innovation challenges? Try Patsnap Eureka!

Application of zp10 in the preparation of anti-Zika virus drugs targeting Zika virus protease

A Zika virus and protease technology, applied in the field of antiviral drugs, can solve the problem of lack of anti-Zika virus data

Active Publication Date: 2021-04-16
MEDICINE & BIOENG INST OF CHINESE ACAD OF MEDICAL SCI
View PDF3 Cites 0 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

To date, only a few natural products of plant origin have been found to be active against the Zika virus protease, IC 50 The range is from 1.3 μM to 56.3 μM, however these compounds lack certain data against Zika virus

Method used

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
View more

Image

Smart Image Click on the blue labels to locate them in the text.
Viewing Examples
Smart Image
  • Application of zp10 in the preparation of anti-Zika virus drugs targeting Zika virus protease
  • Application of zp10 in the preparation of anti-Zika virus drugs targeting Zika virus protease
  • Application of zp10 in the preparation of anti-Zika virus drugs targeting Zika virus protease

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0023] Example 1 ZP10 can significantly inhibit the activity of Zika virus protease

[0024] According to the literature, the present invention uses the tetrapeptide molecule Bz-nKKR-AMC labeled with the fluorescent group AMC as the substrate of ZIKVpro to construct a fluorescence-based screening model. In this model, the concentration of ZIKVpro was 150 nM and the substrate concentration was 50 μM. After testing, the enzyme kinetic parameters (Km=30.51 μM) of Zika virus NS2B-NS3 protease purified in vitro.

[0025] The present invention finds through screening and experiments that ZP10 can significantly inhibit the activity of ZIKVpro, and the structure of ZP10 is as follows: figure 1 shown.

[0026] figure 2 Schematic diagram of the fluorescence-based enzyme kinetic parameter model provided in this example, wherein A is the result of Coomassie brilliant blue staining, SDS-PAGE analysis shows that ZIKVpro is located at about 30KDa, and the purity is higher than 80%, and B...

Embodiment 2

[0028] Example 2 ZP10 Cytotoxicity

[0029] In this embodiment, the cytotoxicity of ZP10 is detected by the CCK-8 kit. The CCK-8 kit is an alternative method for the MTT method, and is a method based on WST-8 (water-soluble tetrazolium salt, chemical name: 2-( Widely used in cell proliferation and cytotoxicity rapid and high-sensitivity detection kit. During detection, the cell supernatant was replaced with cell culture medium containing 10% CCK-8 reagent, and the cells were cultured in a 37°C, 5% CO2 incubator for 1 hour, and the light absorption at 450nm was detected using an EnSpire 2300 multifunctional microplate reader.

[0030] get as Figure 4 The results shown, such as Figure 4 As shown, within 40 μM, ZP10 has no significant effect on cell survival, which means that the antiviral activity of ZP10 has nothing to do with its cytotoxicity, and further combined with Example 1, it can be obtained that the antiviral activity of ZP10 has nothing to do with cytotoxicity. ...

Embodiment 3

[0031] Example 3 Schematic diagram of the effect of ZP10 on Zika virus protease at the transcription and translation levels

[0032] This example detects the impact of ZP10 on the translation level of Zika virus NS2B-NS3 protease by conventional immunoblotting (Western blots) experimental method, and detects Zika virus NS2B-NS3 protease under different concentrations of ZP10 by fluorescent real-time quantitative PCR (qRT-PCR). The changes of NS3 protease at the RNA level, the experimental steps are as follows:

[0033] Western blots: Culture Vero E6 cells with a cell concentration of 2×10 5 / mL, 6-well plates were spread, and after incubation for 24 hours, various concentrations of compound ZP10 (6.25 μM, 12.5 μM, 25 μM) were added, and the final concentration of DMSO was 0.5%. Cells were then infected with Zika virus (MOI=0.05). cells at 37°C, 5% CO 2 The culture was continued for 72 h in the incubator. After 72 hours, the old medium was discarded, and the cells were coll...

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to View More

PUM

PropertyMeasurementUnit
diameteraaaaaaaaaa
Login to View More

Abstract

The present invention relates to the application of ZP10 in the preparation of anti-Zika virus drugs targeting Zika virus protease. By constructing a fluorescence-based screening model, the present invention finds that compound ZP10 has the function of inhibiting Zika virus protease activity. The present invention further studies the cytotoxicity of ZP10 and the affinity with Zika virus protease and finds that ZP10 has higher anti-Zika virus activity, and its IC 50 2.3 μM, EC 50 At the same time, ZP10 itself has low cytotoxicity, has a high affinity with Zika virus protease, and inhibits the replication of Zika virus at the transcription and translation levels, thus having a significant anti-Zika virus effect, which makes ZP10 have a better Anti-Zika virus research and development prospects, can be used to prepare drugs for the prevention or treatment of Zika virus.

Description

technical field [0001] The invention relates to the field of antiviral drugs, in particular to the application of ZP10 in the preparation of anti-Zika virus drugs targeting Zika virus protease Background technique [0002] Zika virus belongs to Flaviviridae, Flavivirus genus, single-stranded positive-sense RNA virus, 20nm in diameter, is an arbovirus transmitted by mosquitoes, the host is not clear, mainly in wild primates and inhabiting trees Mosquitoes, such as Aedes africanus, circulate. [0003] Recent studies have found that Zika virus can cause severe neurological disorders, such as microcephaly in newborns and Guillain-Barre syndrome in adults. However, to date there is no effective vaccine or FDA-approved drug against Zika virus infection. The NS2B-NS3 protease of flaviviruses is critical to the life cycle of Zika virus, so it has become a potential drug target in anti-Zika virus therapy. Natural products are an important source of new drug development worldwide. ...

Claims

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to View More

Application Information

Patent Timeline
no application Login to View More
Patent Type & Authority Patents(China)
IPC IPC(8): A61K31/353A61P31/14
CPCA61K31/353A61P31/14Y02A50/30
Inventor 周金明崔香玲周睿李晓宇岑山
Owner MEDICINE & BIOENG INST OF CHINESE ACAD OF MEDICAL SCI
Who we serve
  • R&D Engineer
  • R&D Manager
  • IP Professional
Why Patsnap Eureka
  • Industry Leading Data Capabilities
  • Powerful AI technology
  • Patent DNA Extraction
Social media
Patsnap Eureka Blog
Learn More
PatSnap group products

Browse by: Latest US Patents, China's latest patents, Technical Efficacy Thesaurus, Application Domain, Technology Topic, Popular Technical Reports.

© 2024 PatSnap. All rights reserved.Legal|Privacy policy|Modern Slavery Act Transparency Statement|Sitemap|About US| Contact US: help@patsnap.com