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

Application of ZP10 in preparation of anti-Zika virus drug taking Zika virus protease as target spot

A Zika virus, 1.ZP10 technology, applied in antiviral agents, pharmaceutical formulas, and resistance to vector-borne diseases, can solve the problem of lack of anti-Zika virus data and other issues

Active Publication Date: 2020-03-06
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 preparation of anti-Zika virus drug taking Zika virus protease as target spot
  • Application of ZP10 in preparation of anti-Zika virus drug taking Zika virus protease as target spot
  • Application of ZP10 in preparation of anti-Zika virus drug taking Zika virus protease as target spot

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 invention uses the tetrapeptide molecule Bz-nKKR-AMC labeled with the fluorescent group AMC as the substrate of ZIKVpro, and constructs a screening model based on fluorescence. In this model, the concentration of ZIKVpro is 150 nM and the concentration of substrate is 50 μM. After testing, the enzyme kinetic parameters of the purified Zika virus NS2B-NS3 protease (Km=30.51μM).

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

[0026] figure 2 The schematic diagram of the fluorescence-based enzyme kinetic parameter model provided in this embodiment, where A is the result of Coomassie brilliant blue staining, SDS-PAGE analysis shows that ZIKVpro is located at about 30KDa with a purity higher than 80%, and B is ZIKVpro's The Mi...

Embodiment 2

[0028] Example 2 ZP10 cytotoxicity

[0029] In this example, the CCK-8 kit was used to detect the cytotoxicity of ZP10. The CCK-8 kit is an alternative to the MTT method and is based on WST-8 (water-soluble tetrazolium salt, chemical name: 2-( 2-Methoxy-4-nitrophenyl)-3-(4-nitrophenyl)-5-(2,4-disulfobenzene)-2H-tetrazole monosodium salt) is widely used in cell proliferation And cytotoxicity rapid and highly sensitive detection kit. During detection, the cell supernatant was replaced with a 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 EnSpire 2300 multifunctional microplate reader was used to detect the light absorption at 450 nm.

[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 in combination with Example 1, the antiviral ac...

Embodiment 3

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

[0032] In this example, the effect of ZP10 on the translation level of Zika virus NS2B-NS3 protease was detected by conventional Western blots, and the effect of ZP10 at different concentrations of ZP10 was detected by fluorescence real-time quantitative PCR (qRT-PCR). The changes in the RNA level of NS3 protease are as follows:

[0033] Western blots: Culture Vero E6 cells at a cell concentration of 2×10 5 / mL, spread a 6-well plate, after 24 hours of incubation, each concentration of compound ZP10 (6.25 μM, 12.5 μM, 25 μM) was added, and the final concentration of DMSO was 0.5%. The cells were subsequently infected with Zika virus (MOI = 0.05). Cell at 37℃, 5% CO 2 Incubate for 72 hours in the incubator. After 72 hours, discard the old medium and collect the cells in a 1.5 mL EP tube. 2×10 3 The cells were collected by centrifugation at g for 5 min, and...

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
Michaelis constantaaaaaaaaaa
Login to View More

Abstract

The invention relates to an application of ZP10 in preparation of an anti-Zika virus drug taking Zika virus protease as a target spot, and researches find that a compound ZP10 has a function of inhibiting the activity of Zika virus protease by constructing a screening model based on fluorescence. According to the invention, the cytotoxicity of ZP10 and the affinity with Zika virus protease are studied, the ZP10 has relatively high Zika virus resisting activity; the IC50 of the compound is 2.3 [mu] M, EC50 is 7.65 [mu] M, meanwhile, the ZP10 is relatively low in cytotoxicity, has relatively high affinity with Zika virus protease, inhibits replication of Zika viruses at transcription and translation levels and further has a remarkable Zika virus resisting effect, so that the ZP10 has a relatively good Zika virus resisting research and development prospect and can be used for preparing medicines for preventing or treating the Zika viruses.

Description

Technical field [0001] The present invention relates to the field of antiviral drugs, in particular to the application of ZP10 in preparing anti-Zika virus drugs targeting Zika virus protease Background technique [0002] Zika virus belongs to the Flaviviridae family, the genus Flavivirus, a single-stranded positive-stranded RNA virus with a diameter of 20nm. It is an arbovirus that is transmitted by mosquitoes. The host is not clear, mainly in wild primates and living on trees. Mosquitoes, such as Aedes African mosquitoes. [0003] Recent studies have found that Zika virus can cause serious neurological diseases, such as neonatal microcephaly and adult Guillain-Barre syndrome. However, there are no effective vaccines and FDA-approved drugs to deal with Zika virus infection. The NS2B-NS3 protease of flavivirus 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 global new dr...

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
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