2-(2-Thienyl)-5,6-dihydroxy-4-carboxypyrimidines as inhibitors of the hepatitis C virus NS5B polymerase: discovery, SAR, modeling, and mutagenesis.
Infections caused by hepatitis C virus (HCV) are a significant world health problem for which novel therapies are in urgent demand. The polymerase of HCV is responsible for the replication of viral RNA. We recently disclosed dihydroxypyrimidine carboxylates 2 as novel, reversible inhibitors of the HCV NS5B polymerase. This series was further developed into 5,6-dihydroxy-2-(2-thienyl)pyrimidine-4-carboxylic acids such as 34 (EC50 9.3 microM), which now show activity in the cell-based HCV replication assay. The structure-activity relationship of these inhibitors is discussed in the context of their physicochemical properties and of the polymerase crystal structure. We also report the results of mutagenesis experiments which support the proposed binding model, which involves pyrophosphate-like chelation of the active site Mg ions.
Koch
Uwe
U
Istituto Di Ricerche Di Biologia Molecolare, P. Angeletti S.p.A. (Merck Research Laboratories, Rome), Via Pontina Km 30,600, 00040 Pomezia, Italy. uwe_koch@merck.com
Attenni
Barbara
B
Malancona
Savina
S
Colarusso
Stefania
S
Conte
Immacolata
I
Di Filippo
Marcello
M
Harper
Steven
S
Pacini
Barbara
B
Giomini
Claudia
C
Thomas
Steven
S
Incitti
Ilario
I
Tomei
Licia
L
De Francesco
Raffaele
R
Altamura
Sergio
S
Matassa
Victor G
VG
Narjes
Frank
F
D000998:Antiviral Agents / Q000138:chemical synthesis* / Q000737:chemistry / Q000494:pharmacology; D001665:Binding Sites; D002460:Cell Line; D002614:Chelating Agents / Q000737:chemistry; D003460:Crystallization; D016174:Hepacivirus / Q000187:drug effects* / Q000201:enzymology*; D006801:Humans; D008782:Methylurea Compounds / Q000138:chemical synthesis* / Q000737:chemistry / Q000494:pharmacology; D008958:Models, Molecular; D016296:Mutagenesis; D011487:Protein Conformation; D011743:Pyrimidines / Q000138:chemical synthesis* / Q000737:chemistry / Q000494:pharmacology; D013329:Structure-Activity Relationship; D013876:Thiophenes / Q000138:chemical synthesis* / Q000737:chemistry / Q000494:pharmacology; D017361:Viral Nonstructural Proteins / Q000037:antagonists & inhibitors* / Q000737:chemistry / Q000235:genetics*; D014779:Virus Replication / Q000187:drug effects; D012324:RNA-Dependent RNA Polymerase
D000998:Antiviral Agents; D002614:Chelating Agents; D008782:Methylurea Compounds; D011743:Pyrimidines; D013876:Thiophenes; D017361:Viral Nonstructural Proteins; C509708:2-(3-((((2-chlorobenzyl)amino)carbonyl)amino)-2-thienyl)-5,6-dihydroxypyrimidine-4-carboxylic acid; C087036:NS-5 protein, hepatitis C virus; D012324:RNA-Dependent RNA Polymerase