Substituted 2,6-bis(benzimidazol-2-yl)pyridines: A novel chemical class of pestivirus inhibitors that targets a hot spot for inhibition of pestivirus replication in the RNA-dependent RNA polymerase

Musiu, Simone; Pürstinger, Gerhard; Stallinger, Sylvia; Vrancken, Robert; Haegeman, Andy; Koenen, F.; Leyssen, Pieter; Froeyen, Mathy; Neyts, Johan; Paeshuyse, Jan

doi:10.1016/j.antiviral.2014.03.010

Cited by 22 publications

(23 citation statements)

References 32 publications

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“…Hepatitis C virus (HCV), the major cause of transfusion-associated hepatitis, also belongs to this family (40). It is well known that NS5B is the viral RNA-dependent RNA polymerase responsible for transcription and replication of the viral genome, and NS5B has been shown to function as part of a larger, membraneassociated replication complex (41,42). Previous studies have shown that inhibitors such as compound VP32947, compound 1457, and BPIP (5 ; with 100 ng of NS5B proteins and 100 ng of poMx1 proteins (WT and mutant) with the RNA templates (C).…”

Section: Discussionmentioning

confidence: 99%

Porcine Mx1 Protein Inhibits Classical Swine Fever Virus Replication by Targeting Nonstructural Protein NS5B

Zhou

Chen

Zhang³

et al. 2018

J Virol

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Mx proteins are interferon (IFN)-induced GTPases that have broad antiviral activity against a wide range of RNA and DNA viruses; they belong to the dynamin superfamily of large GTPases. In this study, we confirmed the anti-classical swine fever virus (CSFV) activity of porcine Mx1 and showed that porcine Mx2 (poMx2), human MxA (huMxA), and mouse Mx1 (mmMx1) also have anti-CSFV activity Small interfering RNA (siRNA) experiments revealed that depletion of endogenous poMx1 or poMx2 enhanced CSFV replication, suggesting that porcine Mx proteins are responsible for the antiviral activity of interferon alpha (IFN-α) against CSFV infection. Confocal microscopy, immunoprecipitation, glutathione -transferase (GST) pulldown, and bimolecular fluorescence complementation (BiFC) demonstrated that poMx1 associated with NS5B, the RNA-dependent RNA polymerase (RdRp) of CSFV. We used mutations in the poMx1 protein to elucidate the mechanism of their anti-CSFV activity and found that mutants that disrupted the association with NS5B lost all anti-CSV activity. Moreover, an RdRp activity assay further revealed that poMx1 undermined the RdRp activities of NS5B. Together, these results indicate that porcine Mx proteins exert their antiviral activity against CSFV by interacting with NS5B. Our previous studies have shown that porcine Mx1 (poMx1) inhibits classical swine fever virus (CSFV) replication and, but the molecular mechanism of action remains largely unknown. In this study, we dissect the molecular mechanism of porcine Mx1 and Mx2 against CSFV Our results show that poMx1 associates with NS5B, the RNA-dependent RNA polymerase of CSFV, resulting in the reduction of CSFV replication. Moreover, the mutants of poMx1 further elucidate the mechanism of their anti-CSFV activities.

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Section: Discussionmentioning

confidence: 99%

Porcine Mx1 Protein Inhibits Classical Swine Fever Virus Replication by Targeting Nonstructural Protein NS5B

Zhou

Chen

Zhang³

et al. 2018

J Virol

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“…They either target a cellular protein/enzyme, i.e., α-glycosidase (which is involved in the maturation of virions [15]), as well as viral encoded enzymes such as the NS3 protease and helicase/NTPase [16], or the NS5B RNA-dependent RNA polymerase (RdRp). Polymerase inhibitors include nucleoside [14] and non-nucleoside inhibitors, such as N-propyl-N-[2-(2H-1,2,4-triazino [5,6-b]indol-3-ylthio)ethyl]-1-propanamine (VP32947) [17], a thiazole urea derivative [18], a cyclic urea derivative [19], imidazo-pyridines (BPIP) [20], ethyl 2-methylimidazo [1,2-a]pyrrolo [2,3-c]pyridin-8-carboxylate (AG110) [21], pyraz olotriazolopyrimidinamine (LZ37) [22], 2-(2-benzimidazolyl)-5-[4-(2-imidazolino)phenyl]furan (DB772) [23], 5,6-dimethoxy-1-indanone [24,25], 2-phenylbenzimidazole [26], substituted 2,6-bis (benzimidazol-2-yl)pyridines [27], benzimidazole derivative [28], and arylazoenamine derivatives [29].…”

Section: Introductionmentioning

confidence: 99%

“…BVDV strains that are resistant to the majority of these non-nucleosidic RdRp inhibitors all carry mutations in the fingertip domain of the viral RdRp. However, most of these inhibitors do not inhibit the in vitro activity of the recombinant viral polymerase but are able to inhibit the activity of the BVDV replicase complex (RC), in a dose dependent manner [20][21][22]27,30]. The fingertip domain of the polymerase is crucial for the function of the polymerase and the viral RC.…”

Section: Introductionmentioning

confidence: 99%

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Quinolinecarboxamides Inhibit the Replication of the Bovine Viral Diarrhea Virus by Targeting a Hot Spot for the Inhibition of Pestivirus Replication in the RNA-Dependent RNA Polymerase

et al. 2020

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The bovine viral diarrhea virus (BVDV), a pestivirus from the family of Flaviviridae is ubiquitous and causes a range of clinical manifestations in livestock, mainly cattle. Two quinolinecarboxamide analogues were identified in a CPE-based screening effort, as selective inhibitors of the in vitro bovine viral diarrhea virus (BVDV) replication, i.e., TO505-6180/CSFCI (average EC50 = 0.07 µM, SD = 0.02 µM, CC50 > 100 µM) and TO502-2403/CSFCII (average EC50 = 0.2 µM, SD = 0.06 µM, CC50 > 100 µM). The initial antiviral activity observed for both hits against BVDV was corroborated by measuring the inhibitory effect on viral RNA synthesis and the production of infectious virus. Modification of the substituents on the quinolinecarboxamide scaffold resulted in analogues that proved about 7-fold more potent (average EC50 = 0.03 with a SD = 0.01 µM) and that were devoid of cellular toxicity, for the concentration range tested (SI = 3333). CSFCII resistant BVDV variants were selected and were found to carry the F224P mutation in the viral RNA-dependent RNA polymerase (RdRp), whereas CSFCI resistant BVDV carried two mutations in the same region of the RdRp, i.e., N264D and F224Y. Likewise, molecular modeling revealed that F224P/Y and N264D are located in a small cavity near the fingertip domain of the pestivirus polymerase. CSFC-resistant BVDV proved to be cross-resistant to earlier reported pestivirus inhibitors (BPIP, AG110, LZ37, and BBP) that are known to target the same region of the RdRp. CSFC analogues did not inhibit the in vitro activity of recombinant BVDV RdRp but inhibited the activity of BVDV replication complexes (RCs). CSFC analogues likely interact with the fingertip of the pestivirus RdRp at the same position as BPIP, AG110, LZ37, and BBP. This indicates that this region is a “hot spot” for the inhibition of pestivirus replication.

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“…CSFV possesses a single-stranded, positive-sense RNA genome. The virus-encoded RNA-dependent RNA polymerase (RdRp) is the central component in the replication of RNA viruses, including CSFV (Hansen et al 1997), which is therefore an important target for antiviral drug development (Baginski et al 2000;Musiu et al 2014;Paeshuyse et al 2006;Sun et al 2003;Vrancken et al 2009a;Vrancken et al 2009b;Vrancken et al 2008).…”

Section: R a F T Introductionmentioning

confidence: 99%

Episodic adaptive diversification of classical swine fever virus RNA-dependent RNA polymerase NS5B

Yang

2015

Can. J. Microbiol.

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Classical swine fever virus (CSFV) is the pathogen that causes a highly infectious disease of pigs and has led to disastrous losses to pig farms and related industries. The RNA-dependent RNA polymerase (RdRp) NS5B is a central component of the replicase complex (RC) in some single-stranded RNA viruses, including CSFV. On the basis of genetic variation, the CSFV RdRps could be clearly divided into 2 major groups and a minor group, which is consistent with the phylogenetic relationships and virulence diversification of the CSFV isolates. However, the adaptive signature underlying such an evolutionary profile of the polymerase and the virus is still an interesting open question. We analyzed the evolutionary trajectory of the CSFV RdRps over different timescales to evaluate the potential adaptation. We found that adaptive selection has driven the diversification of the RdRps between, but not within, CSFV major groups. Further, the major adaptive divergence-related sites are located in the surfaces relevant to the interaction with other component(s) of RC and the entrance and exit of the template-binding channel. These results might shed some light on the nature of the RdRp in virulence diversification of CSFV groups.

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Substituted 2,6-bis(benzimidazol-2-yl)pyridines: A novel chemical class of pestivirus inhibitors that targets a hot spot for inhibition of pestivirus replication in the RNA-dependent RNA polymerase

Cited by 22 publications

References 32 publications

Porcine Mx1 Protein Inhibits Classical Swine Fever Virus Replication by Targeting Nonstructural Protein NS5B

Porcine Mx1 Protein Inhibits Classical Swine Fever Virus Replication by Targeting Nonstructural Protein NS5B

Quinolinecarboxamides Inhibit the Replication of the Bovine Viral Diarrhea Virus by Targeting a Hot Spot for the Inhibition of Pestivirus Replication in the RNA-Dependent RNA Polymerase

Episodic adaptive diversification of classical swine fever virus RNA-dependent RNA polymerase NS5B

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