title: Biochemical Characterization of the Active Anti-Hepatitis C Virus Metabolites of 2,6-Diaminopurine Ribonucleoside Prodrug Compared to Sofosbuvir and BMS-986094.
abstract: Ribonucleoside analog inhibitors (rNAI) target the hepatitis C virus (HCV) RNA-dependent RNA polymerase nonstructural protein 5B (NS5B) and cause RNA chain termination. Here, we expand our studies on β-d-2'-C-methyl-2,6-diaminopurine-ribonucleotide (DAPN) phosphoramidate prodrug 1 (PD1) as a novel investigational inhibitor of HCV. DAPN-PD1 is metabolized intracellularly into two distinct bioactive nucleoside triphosphate (TP) analogs. The first metabolite, 2'-C-methyl-GTP, is a well-characterized inhibitor of NS5B polymerase, whereas the second metabolite, 2'-C-methyl-DAPN-TP, behaves as an adenosine base analog. In vitro assays suggest that both metabolites are inhibitors of NS5B-mediated RNA polymerization. Additional factors, such as rNAI-TP incorporation efficiencies, intracellular rNAI-TP levels, and competition with natural ribonucleotides, were examined in order to further characterize the potential role of each nucleotide metabolite in vivo Finally, we found that although both 2'-C-methyl-GTP and 2'-C-methyl-DAPN-TP were weak substrates for human mitochondrial RNA (mtRNA) polymerase (POLRMT) in vitro, DAPN-PD1 did not cause off-target inhibition of mtRNA transcription in Huh-7 cells. In contrast, administration of BMS-986094, which also generates 2'-C-methyl-GTP and previously has been associated with toxicity in humans, caused detectable inhibition of mtRNA transcription. Metabolism of BMS-986094 in Huh-7 cells leads to 87-fold higher levels of intracellular 2'-C-methyl-GTP than DAPN-PD1. Collectively, our data characterize DAPN-PD1 as a novel and potent antiviral agent that combines the delivery of two active metabolites.
authors.affiliation: Center for AIDS Research, Laboratory of Biochemical Pharmacology, Department of Pediatrics, Emory University School of Medicine, Atlanta, Georgia, USA.
authors.forename: Maryam
authors.initials: M
authors.lastname: Ehteshami
authors.affiliation: Center for AIDS Research, Laboratory of Biochemical Pharmacology, Department of Pediatrics, Emory University School of Medicine, Atlanta, Georgia, USA.
authors.forename: Sijia
authors.initials: S
authors.lastname: Tao
authors.affiliation: Center for AIDS Research, Laboratory of Biochemical Pharmacology, Department of Pediatrics, Emory University School of Medicine, Atlanta, Georgia, USA.
authors.forename: Tugba
authors.initials: T
authors.lastname: Ozturk
authors.affiliation: Center for AIDS Research, Laboratory of Biochemical Pharmacology, Department of Pediatrics, Emory University School of Medicine, Atlanta, Georgia, USA.
authors.forename: Longhu
authors.initials: L
authors.lastname: Zhou
authors.affiliation: Center for AIDS Research, Laboratory of Biochemical Pharmacology, Department of Pediatrics, Emory University School of Medicine, Atlanta, Georgia, USA.
authors.forename: Jong Hyun
authors.initials: JH
authors.lastname: Cho
authors.affiliation: Center for AIDS Research, Laboratory of Biochemical Pharmacology, Department of Pediatrics, Emory University School of Medicine, Atlanta, Georgia, USA.
authors.forename: Hongwang
authors.initials: H
authors.lastname: Zhang
authors.affiliation: Center for AIDS Research, Laboratory of Biochemical Pharmacology, Department of Pediatrics, Emory University School of Medicine, Atlanta, Georgia, USA.
authors.forename: Sheida
authors.initials: S
authors.lastname: Amiralaei
authors.affiliation: Center for AIDS Research, Laboratory of Biochemical Pharmacology, Department of Pediatrics, Emory University School of Medicine, Atlanta, Georgia, USA.
authors.forename: Jadd R
authors.initials: JR
authors.lastname: Shelton
authors.affiliation: Center for AIDS Research, Laboratory of Biochemical Pharmacology, Department of Pediatrics, Emory University School of Medicine, Atlanta, Georgia, USA.
authors.forename: Xiao
authors.initials: X
authors.lastname: Lu
authors.affiliation: Center for AIDS Research, Laboratory of Biochemical Pharmacology, Department of Pediatrics, Emory University School of Medicine, Atlanta, Georgia, USA.
authors.forename: Ahmed
authors.initials: A
authors.lastname: Khalil
authors.affiliation: Center for AIDS Research, Laboratory of Biochemical Pharmacology, Department of Pediatrics, Emory University School of Medicine, Atlanta, Georgia, USA.
authors.forename: Robert A
authors.initials: RA
authors.lastname: Domaoal
authors.affiliation: Center for AIDS Research, Laboratory of Biochemical Pharmacology, Department of Pediatrics, Emory University School of Medicine, Atlanta, Georgia, USA.
authors.forename: Richard A
authors.initials: RA
authors.lastname: Stanton
authors.affiliation: Center for AIDS Research, Laboratory of Biochemical Pharmacology, Department of Pediatrics, Emory University School of Medicine, Atlanta, Georgia, USA.
authors.forename: Justin E
authors.initials: JE
authors.lastname: Suesserman
authors.affiliation: Cocrystal Pharma, Inc., Tucker, Georgia, USA.
authors.forename: Biing
authors.initials: B
authors.lastname: Lin
authors.affiliation: Cocrystal Pharma, Inc., Tucker, Georgia, USA.
authors.forename: Sam S
authors.initials: SS
authors.lastname: Lee
authors.affiliation: Center for AIDS Research, Laboratory of Biochemical Pharmacology, Department of Pediatrics, Emory University School of Medicine, Atlanta, Georgia, USA.
authors.forename: Franck
authors.initials: F
authors.lastname: Amblard
authors.affiliation: Cocrystal Pharma, Inc., Tucker, Georgia, USA.
authors.forename: Tony
authors.initials: T
authors.lastname: Whitaker
authors.affiliation: Cocrystal Pharma, Inc., Tucker, Georgia, USA.
authors.forename: Steven J
authors.initials: SJ
authors.lastname: Coats
authors.affiliation: Center for AIDS Research, Laboratory of Biochemical Pharmacology, Department of Pediatrics, Emory University School of Medicine, Atlanta, Georgia, USA rschina@emory.edu.
authors.forename: Raymond F
authors.initials: RF
authors.lastname: Schinazi
mesh_terms: D000241:Adenosine / Q000031:analogs & derivatives* / Q000494:pharmacology; D000998:Antiviral Agents / Q000494:pharmacology*; D002460:Cell Line; D012321:DNA-Directed RNA Polymerases / Q000378:metabolism; D006157:Guanosine Monophosphate / Q000031:analogs & derivatives* / Q000494:pharmacology; D016174:Hepacivirus / Q000187:drug effects*; D006526:Hepatitis C / Q000188:drug therapy*; D006801:Humans; D011355:Prodrugs / Q000494:pharmacology*; D012313:RNA / Q000378:metabolism; D000077278:RNA, Mitochondrial; D012367:RNA, Viral / Q000378:metabolism; D012263:Ribonucleosides / Q000378:metabolism; D000069474:Sofosbuvir / Q000494:pharmacology*; D014158:Transcription, Genetic / Q000187:drug effects; D017361:Viral Nonstructural Proteins / Q000378:metabolism; D014779:Virus Replication / Q000187:drug effects
chemical_list: D000998:Antiviral Agents; D011355:Prodrugs; D000077278:RNA, Mitochondrial; D012367:RNA, Viral; D012263:Ribonucleosides; D017361:Viral Nonstructural Proteins; C033854:2-aminoadenosine; C553340:BMS-986094; D012313:RNA; D006157:Guanosine Monophosphate; D012321:DNA-Directed RNA Polymerases; D000241:Adenosine; D000069474:Sofosbuvir