Toll-like receptor 2 antagonists identified through virtual screening and experimental validation

5 years ago

Toll-like receptor 2 antagonists identified through virtual screening and experimental validation

Dhanusha Yesudhas, Rajiv Gandhi Govindaraj, Hyeon-Jun Shin, Suresh Panneerselvam, Hyuk-Kwon Kwon, Kyoung Tai No, Asma Achek, Sangdun Choi, Jiwon Choi, Prasannavenkatesh Durai

Toll-like receptor 2 (TLR2) antagonists are key therapeutic targets because they inhibit several inflammatory diseases caused by surplus TLR2 activation. In this study, we identified two novel nonpeptide TLR2 antagonists, C11 and C13, through pharmacophore-based virtual screening. At 10 μm, the level of interleukin (IL)-8 inhibition by C13 and C11 in human embryonic kidney TLR2 overexpressing cells was comparable to the commercially available TLR2 inhibitor CU-CPT22. In addition, C11 and C13 acted in mouse macrophage-like RAW 264.7 cells as TLR2-specific inhibitors and did not suppress the tumor necrosis factor-α induction by TLR3 and TLR4 activators. Moreover, the two identified compounds bound directly to the human recombinant TLR2 ectodomain, during surface plasmon resonance analysis, and did not affect cell viability in a 3-(4,5-dimethylthiazol-2-yl)-5(3-carboxymethonyphenol)-2-(4-sulfophenyl)-2H-tetrazolium assay. In total, two virtually screened molecules, C11 and C13, were experimentally proven to be effective as TLR2 antagonists, and thus will provide new insights into the structure of TLR2 antagonists, and pave the way for the development of TLR2-targeted drug molecules. By combining in silico predictions and in vitro confirmations, the TLR2 antagonists C13 and C11 were identified. The two novel TLR2 inhibitors are TLR2 specific and suppressed the cytokines in both human HEK293-hTLR2 (interlukein-8 inhibition) and mouse RAW 264.7 (tumor necrosis factor-α inhibition) cells.

Publisher URL: http://onlinelibrary.wiley.com/resolve/doi

DOI: 10.1111/febs.14124