A pyrazolotriazolopyrimidinamine inhibitor of bovine viral diarrhea virus replication that targets the viral RNA-dependent RNA polymerase.
[7-[3-(1,3-Benzodioxol-5-yl)propyl]-2-(2-furyl)-7H-pyrazolo[4,3-e][1,2,4]triazolo[1,5-c]pyrimidin-5-amine] (LZ37) was identified as a selective inhibitor of in vitro bovine viral diarrhea virus (BVDV) replication. The EC(50) values for inhibition of BVDV-induced cytopathic effect (CPE) formation, viral RNA synthesis and production of infectious virus were 4.3+/-0.7microM, 12.9+/-1microM and 5.8+/-0.6microM, respectively. LZ37 proved inactive against the hepatitis C virus and the flavivirus yellow fever. LZ37 inhibits BVDV replication at a time point that coincides with the onset of intracellular viral RNA synthesis. Drug-resistant mutants carried the F224Y mutation in the viral RNA-dependent RNA polymerase (RdRp). LZ37 showed cross-resistance with the imidazopyrrolopyridine AG110 [which selects for the E291G drug resistance mutation] as well as with the imidazopyridine BPIP [which selects for the F224S drug-resistant mutation]. LZ37 did not inhibit the in vitro activity of purified recombinant BVDV RdRp. Molecular modelling revealed that F224 is located near the tip of the finger domain of the RdRp. Docking of LZ37 in the crystal structure of the BVDV RdRp revealed several potential contacts including: (i) hydrophobic contacts of LZ37 with A221, A222, G223, F224 and A392; (ii) a stacking interaction between F224 side chain and the ring system of LZ37 and (iii) a hydrogen bond between the amino function of LZ37 and the O backbone atom of A392. It is concluded that LZ37 interacts with the same binding site as BPIP or VP32947 at the top of the finger domain of the polymerase that is a "hot spot" for inhibition of pestivirus replication.
Rega Institute for Medical Research, Katholieke Universiteit Leuven, Belgium. Jan.Paeshuyse@rega.kuleuven.be
Jan
J
Paeshuyse
Carine
C
Letellier
Matheus
M
Froeyen
Hélène
H
Dutartre
Robert
R
Vrancken
Bruno
B
Canard
Erik
E
De Clercq
Alain
A
Gueiffier
Jean-Claude
JC
Teulade
Piet
P
Herdewijn
Gerhard
G
Puerstinger
Frank
F
Koenen
Pierre
P
Kerkhofs
Pier Giovanni
PG
Baraldi
Johan
J
Neyts
D019943:Amino Acid Substitution / Q000235:genetics; D000818:Animals; D000998:Antiviral Agents / Q000737:chemistry / Q000494:pharmacology*; D052117:Benzodioxoles / Q000737:chemistry / Q000494:pharmacology*; D002417:Cattle; D002460:Cell Line; D030241:Diarrhea Virus 1, Bovine Viral / Q000187:drug effects* / Q000502:physiology; D030242:Diarrhea Virus 2, Bovine Viral / Q000187:drug effects* / Q000502:physiology; D024882:Drug Resistance, Viral; D016174:Hepacivirus / Q000187:drug effects; D020128:Inhibitory Concentration 50; D008958:Models, Molecular; D020125:Mutation, Missense; D012324:RNA-Dependent RNA Polymerase / Q000737:chemistry / Q000235:genetics; D014230:Triazoles / Q000737:chemistry / Q000494:pharmacology*; D014764:Viral Proteins / Q000235:genetics; D014779:Virus Replication / Q000187:drug effects*; D015005:Yellow fever virus / Q000187:drug effects
C541938:7-(3-(1,3-benzodioxol-5-yl)propyl)-2-(2-furyl)-7H-pyrazolo(4,3-e)(1,2,4)triazolo(1,5-c)pyrimidin-5-amine; D000998:Antiviral Agents; D052117:Benzodioxoles; D014230:Triazoles; D014764:Viral Proteins; D012324:RNA-Dependent RNA Polymerase