研究目的
To develop small molecule inhibitors targeting the flavivirus envelope protein, E, to combat dengue, Zika, and other flavivirus pathogens without the risk of antibody-dependent enhancement of infection and disease.
研究成果
The study provides proof of concept for the development of direct-acting antivirals that can block E-mediated membrane fusion of multiple flavivirus pathogens. The cyanohydrazone inhibitors, particularly JBJ-01-162-04, show promise as broad-spectrum antivirals against flaviviruses, with improved in vivo properties and reduced cytotoxicity.
研究不足
The study acknowledges the potential for nonspecific, E-independent mechanisms, including colloidal compound aggregation or other pan-assay interference (PAINS) properties, to contribute to antiviral activity. Additionally, the modest effect observed in vivo may be due to high levels of plasma-protein binding, limiting the compound's antiviral effect.
1：Experimental Design and Method Selection:
The study utilized both phenotypic and target-based approaches to discover small molecules that bind to the DENV prefusion E dimer (E2) on the virion surface and block infection by preventing E-mediated membrane fusion.
2：Sample Selection and Data Sources:
Viral stocks of dengue virus serotype 2 strain New Guinea C (DENV2 NGC), dengue virus serotype 2 strain S221 (DENV2 S221), Zika virus (ZIKV) strain PF-251013-18, and Japanese encephalitis virus (JEV) vaccine strain SA14-14-2 were used.
3：List of Experimental Equipment and Materials:
Instruments included a Waters Acquity UPLC/MS system, Octet RED384 system (ForteBio), and Typhoon FLA 9500 (GE Healthcare Life Sciences). Materials included liposomes composed of DOPC, DOPE, PI, BMP, and cholesterol.
4：Experimental Procedures and Operational Workflow:
The viral infectivity assay was designed to ensure that the antiviral activity observed is due to an effect on viral entry. Compound treatment was limited to a preincubation with the inoculum and the initial 1 h infection, after which nonadsorbed virus and compound were washed away. After 24 h, the yield of infectious virus secreted to the culture supernatant was quantified.
5：Data Analysis Methods:
Data were analyzed using nonlinear regression for IC90 and CC50 determinations, and statistical analysis was performed using Prism (GraphPad Software).
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