3-(imidazo[1,2- a :5,4- b ′]dipyridin-2-yl)aniline inhibits pestivirus replication by targeting a hot spot drug binding pocket in the RNA-dependent RNA polymerase

Musiu, Simone; Leyssen, Pieter; Froeyen, Mathy; Chezal, Jean‐Michel; Neyts, Johan; Paeshuyse, Jan

doi:10.1016/j.antiviral.2016.03.007

Cited by 12 publications

(10 citation statements)

References 31 publications

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“…Assays were carried out, as described previously [20][21][22]27,30]. In brief, MDBK cells were seeded at a density of 5 × 10 3 per well in 96-well cell culture plates, in MEM containing 5% FBS.…”

Section: Cytotoxicity Assaymentioning

confidence: 99%

“…2.6. Isolation of TO502-2403 res /CSFCII res TO505-6180 res /CSFCI res BVDV CSFC-resistant (CSFC res ) virus was selected, as previously described [20][21][22]27,30]. In brief, CSFC res BVDV was generated by culturing wild-type BVDV in MDBK cells, in the presence of increasing concentrations of the compound in a 48-well plate.…”

Section: D Qsar Modelmentioning

confidence: 99%

“…RT-quantitative PCR (RT-qPCR) was performed, as previously described [20][21][22]27,30]. In brief, a 25-µL RT-qPCR reaction was composed of a 12.5 µL 2 × reaction buffer (Eurogentec, Seraing, Belgium), 6.3 µL H 2 O, 300 nmol/L forward and reverse primer, 300 nmol/L TaqMan probe and 5 µL total cellular or viral RNA extract.…”

Section: Rt-qpcrmentioning

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%

“…The fingertip domain of the polymerase is crucial for the function of the polymerase and the viral RC. This domain is, thus, apparently a "hot spot" binding site for selective inhibitors of pestivirus replication [20][21][22]27,30].…”

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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