Chemical array system, a platform to identify novel hepatitis B virus entry inhibitors targeting sodium taurocholate cotransporting polypeptide

Abstract

Current anti-hepatitis B virus (HBV) agents including interferons and nucleos(t)ide analogs efficiently suppress HBV infection. However, as it is difficult to eliminate HBV from chronically infected liver, alternative anti-HBV agents targeting a new molecule are urgently needed. In this study, we applied a chemical array to high throughput screening of small molecules that interacted with sodium taurocholate cotransporting polypeptide (NTCP), an entry receptor for HBV. From approximately 30,000 compounds, we identified 74 candidates for NTCP interactants, and five out of these were shown to inhibit HBV infection in cell culture. One of such compound, NPD8716, a coumarin derivative, interacted with NTCP and inhibited HBV infection without causing cytotoxicity. Consistent with its NTCP interaction capacity, this compound was shown to block viral attachment to host hepatocytes. NPD8716 also prevented the infection with hepatitis D virus, but not hepatitis C virus, in agreement with NPD8716 specifically inhibiting NTCP-mediated infection. Analysis of derivative compounds showed that the anti-HBV activity of compounds was apparently correlated with the affinity to NTCP and the capacity to impair NTCP-mediated bile acid uptake. These results are the first to show that the chemical array technology represents a powerful platform to identify novel viral entry inhibitors.

Troglitazone Impedes the Oligomerization of Sodium Taurocholate Cotransporting Polypeptide and Entry of Hepatitis B Virus Into Hepatocytes

“…The preS1-NTCP interaction appears to trigger internalization by an as-yet unknown mechanism (Karayiannis, 2017). To date, a series of HBV entry inhibitors has been reported, including a lipopeptide (Myrcludex-B), bile acids, FDA-approved drugs (cyclosporin A, irbesartan, ezetimibe, and rapamycin), and additional compounds (including vanitaracins, proanthocyanidin, SCY995, NPD8716, and WL4; Lucifora et al, 2013; Blanchet et al, 2014; Iwamoto et al, 2014; König et al, 2014; Ni et al, 2014; Nkongolo et al, 2014; Watashi et al, 2014; Yan et al, 2014; Kaneko et al, 2015, 2018; Ko et al, 2015; Tsukuda et al, 2015, 2017; Veloso Alves Pereira et al, 2015; Wang et al, 2015; Donkers et al, 2017; Shimura et al, 2017; Passioura et al, 2018; Saso et al, 2018; Song et al, 2018). These compounds target the host NTCP protein or HBV particles themselves, inhibiting the attachment of HBV to the host cell surface.…”

Concept of Viral Inhibitors via NTCP

“…85 By applying this technology to the screening of small molecules that interact with recombinant NTCP protein, NPD8716, a coumarin derivative, was identified (from among $30,000 compounds) based on its ability to interact directly with NTCP. 86 This compound was shown to inhibit HBV infection by interfering with viral preS1 attachment to hepatocytes in a cell culture system. Coumarin derivatives are expected to be potential candidates in drug development, given that this class of compounds has already been shown to be safe for clinical use.…”

Phytotherapy and Liver Disease

“…It is assumed that 4-MU slows down progression of hepatitis into cirrhosis and liver cancer [91]. Research into coumarin metabolism proved that rats are not suitable model for research into toxicity of coumarin for humans [92]. Coumarins generally display multiple biological activities: anticoagulative, estrogenic, photosensitive, antimicrobial, vasodilatative, antiviral against HBV and HDV [93] and HCV, and molluscicidal activity; they act as anthelmintic (in digestive system) and as sedatives and hypnotics and also indicate analgesic and hypothermal effects.…”