Abstract
Chikungunya virus (CHIKV) infection and subsequent high patient morbidity is a global threat. The present study aimed to identify the potent antiviral agent against Chikungunya virus, with minimum in vitro cytotoxicity. CHIKV nsP4 3D structure was determined using the I-TASSER server followed by its refinement and pocket determination. Furthermore, high-throughput molecular docking was employed to identify candidate CHIKV nsP4 inhibitors in a library containing 214 compounds. The top ranked compound was evaluated further with various assays, including cytotoxicity, antiviral activity, time of drug addition, viral entry attachment, and microneutralization assays. High-throughput computational screening indicated silibinin to have the best interaction with CHIKV nsP4 protein, immature and mature glycoproteins with highest negative free binding energy, − 5.24 to − 5.86 kcal/mol, and the lowest inhibitory constant, 50.47 to 143.2 µM. Further in vitro analysis demonstrated silibinin could exhibit statistically significant (p < 0.05) dose-dependent anti-CHIKV activity within 12.5–100-µM concentrations with CC50 as 50.90 µM. In total, 50 µM silibinin interfered with both CHIKV attachment (75%) and entry (82%) to Vero cells. Time of addition assay revealed silibinin interfered with late phase of the CHIKV replication cycle. Microneutralization assay revealed that silibinin could inhibit clearing of 50% Vero cell monolayer caused by CHIKV-induced CPE at a minimum dose of 25 µM. These data indicated silibinin to be a promising candidate drug against CHIKV infection.
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Funding
This study was partly funded by Department of Biotechnology, Government of West Bengal, India [No.132-BT (Estt.) /RD-1/10].
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11626_2022_666_MOESM1_ESM.doc
Supplementary file1 Different grid coordinates x, y, z; spacing; grid box size and location of immature and mature glycoprotein structure considered for autodock analysis. (DOC 29 KB)
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Supplementary file2 Binding energy of CHIKV nsP4 protein with selected 214 compounds considered for primary screening by iGEMDOCK v.2. (DOC 257 KB)
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Dutta, S.K., Sengupta, S. & Tripathi, A. In silico and in vitro evaluation of silibinin: a promising anti-Chikungunya agent. In Vitro Cell.Dev.Biol.-Animal 58, 255–267 (2022). https://doi.org/10.1007/s11626-022-00666-x
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DOI: https://doi.org/10.1007/s11626-022-00666-x