2′-Deoxy-2′-spirocyclopropylcytidine Revisited: A New and Selective Inhibitor of the Hepatitis C Virus NS5B Polymerase

Chronic infection with hepatitis C virus (HCV) is a major global health burden and is associated with an increased risk of liver cirrhosis and hepatocellular carcinoma. Current therapy for HCV infection has limited efficacy, particularly against genotype 1 virus, and is hampered by a range of adverse effects. Therefore, there is a clear unmet medical need for efficacious and safe direct antiviral drugs for use in combination with current treatments to increase cure rates and shorten treatment times. The broad genotypic coverage achievable with nucleosides or nucleotides and the high genetic barrier to resistance of these compounds observed in vitro and in vivo suggest that this class of inhibitors could be a valuable component of future therapeutic regimens. Here, we report the in vitro inhibitory activity and mode of action of 2-deoxy-2-spirocyclopropylcytidine (TMC647078), a novel and potent nucleoside inhibitor of the HCV NS5B RNA-dependent RNA polymerase that causes chain termination of the nascent HCV RNA chain. In vitro combination studies with a protease inhibitor resulted in additive efficacy in the suppression of HCV RNA replication, highlighting the potential for the combination of these two classes in the treatment of chronic HCV infection. No cytotoxic effects were observed in various cell lines. Biochemical studies indicated that TMC647078 is phosphorylated mainly by deoxycytidine kinase (dCK) without inhibiting the phosphorylation of the natural substrate, and high levels of triphosphate were observed in Huh7 cells and in primary hepatocytes in vitro. TMC647078 is a potent novel nucleoside inhibitor of HCV replication with a promising in vitro virology and biology profile.Infection with hepatitis C virus (HCV), the causative agent of hepatitis C, is an important global health burden, with an estimated 120 to 170 million persons chronically infected (7,11). Chronic HCV infection can lead to liver cirrhosis and hepatocellular carcinoma and is the leading cause of liver transplantation (10). The virus is transmitted mainly via bloodblood contact, and spontaneous virus clearance has been estimated to be achieved for 26% of infected subjects (29).HCV is a member of the Flaviviridae family of viruses in the genus Hepacivirus, comprising at least six major genotypes and multiple subtypes. The 9.6-kb positive-sense, single-stranded RNA genome of HCV encodes four structural proteins (the core protein, the envelope glycoproteins E1 and E2, and p7) and six nonstructural proteins responsible for viral replication (NS2, NS3, NS4A, NS4B, NS5A, and NS5B). The nonstructural protein NS5B is an RNA-dependent RNA polymerase (RdRp) responsible for the amplification of the HCV RNA and the assembly of the replicase complex at the endoplasmic reticulum membrane (3, 25).Currently, HCV patients are treated with a combination of weekly pegylated alpha interferon (IFN-␣) injections and twice-daily ribavirin, which has considerable tolerability issues (28) and results in a sustained viral response in only 40 to 50% of patient...

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“…We recently described the synthesis and anti-HCV activity of TMC647078, a 2Ј-deoxy-2Ј-spirocyclopropylcytidine (14).…”

Section: Discussionmentioning

confidence: 99%

Antiviral Activity and Mode of Action of TMC647078, a Novel Nucleoside Inhibitor of the Hepatitis C Virus NS5B Polymerase

Berke¹,

Vijgen²,

Lachau‐Durand³

et al. 2011

Antimicrob Agents Chemother

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“…2′-Deoxy-2′-spirocyclopropylcytidine (1, EC 50 = 7.3 μM, Figure 1) was discovered as a new member of the class of 2′-modified nucleoside derivatives and showed potent anti-HCV activity. 13 A 2′-spiroetheruridine analogue (2, EC 50 = 14.9 μM, Figure 1) and its phosphoramidate prodrug (3, EC 50 = 20.6 μM, Figure 1) exhibited medium activity against HCV. 14 This may suggest that the 2′-α-C-alkyl in nucleosides/ nucleotides might be tolerated and that the 2′-α-OH or 2′-α-F may not be essential for nucleosides/nucleotides to exhibit anti-HCV activity.…”

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confidence: 99%

Discovery of 2′-α-C-Methyl-2′-β-C-fluorouridine Phosphoramidate Prodrugs as Inhibitors of Hepatitis C Virus

Zeng

Zhang

Nie

et al. 2016

ACS Med. Chem. Lett.

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2′-α-C-Methyl-2′-β-C-fluorouridine and its phosphoramidate prodrugs were synthesized and evaluated for their inhibitory activity against HCV. The structure− activity relationship analysis of the phosphoramidate moiety found that 17m, 17q, and 17r exhibit potent activities against HCV, with EC 50 values of 1.82 ± 0.19, 0.88 ± 0.12, and 2.24 ± 0.22 μM, respectively. The docking study revealed that the recognition of the 2′-β-F by Arg158, 3′-OH by N291, and the Watson−Crick pairing with the template allowed 23 to form the in-line conformation necessary for its incorporation into the viral RNA chain. KEYWORDS: hepatitis C virus, phosphoramidate prodrugs, docking, inhibitor, Hint 1 H epatitis C virus (HCV) is a small, single-stranded positive-sense RNA virus that was explicitly identified in 1989. 1 The 9.6 kb RNA genome shares similarities with the genomes of flaviviruses and pestiviruses. 2 HCV infection is a global health problem that impacts approximately 180 million individuals, of which approximately 150 million people may proceed to develop chronic liver disease. A significant percentage of patients may ultimately progress to liver cirrhosis and eventually develop hepatocellular carcinoma. 3 HCV infection is also the primary reason for liver transplants among adults. The therapy for HCV infection has long been regular injections of pegylated α-interferon with the daily oral administration of ribavirin (RBV). 4 This standard of care (SOC) functions by enhancing the host immune system rather than acting directly on the virus. However, HCV patients undergoing this therapy may also suffer significant adverse effects, including fatigue, hemolytic anemia, depression, and flulike symptoms, which are poorly tolerated. In 2011, boceprevir and telaprevir, as HCV protease inhibitors, became available to treat HCV infection with genotype 1 in combination with ribavirin and pegylated interferon. 5 In 2013, two new directacting antivirals (DAAs), simeprevir and sofosbuvir, were approved by the FDA, which significantly enhanced the available armory. 6 However, the high cost for a course of treatment with sofosbuvir creates a considerable economic burden for patients. 7 Hence, the search for novel DAAs that are safe and effective remains a necessary endeavor.The HCV NS5B RNA-dependent RNA polymerase is a key enzyme in the replication of the virus. Therefore, it has become an attractive target for the development of small molecule inhibitors of viral replication. Nucleoside and nucleotide inhibitors of HCV (NIs) are a class of DAAs that show broad activity across HCV genotypes and a high barrier to the emergence of viral resistance. 8 Structural modifications have been made to both the base and ribose sugar portions of a ribonucleoside to develop potent and selective anti-HCV inhibitors. 9 Structural modifications of the ribose sugar portion focused primarily on the 1′-, 2′-, and 4′-positions. Structural modifications of the 2′-position mainly consisted of introducing an alkyl or including a methyl at the β-position ...

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“…15,16 They were complemented with the spirocyclic 2'-deoxy-2'-spirocyclopropyl and the 2'-deoxy-2'-spirooxetane moeity. 17,18,19 Recently, the 2'-α-fluoro-2'-β-chloro derivative was also reported 20 ( Figure 1). These 2'-variants not only need to mimic the interaction between the polymerase and the 2'-α-OH group present in natural ribonucleotides, they also must conform to the steric limitations of the corresponding 2'-region of the polymerase to result in proper inhibition thereof.…”

Section: Introductionmentioning

confidence: 92%

Discovery of 1-((2R,4aR,6R,7R,7aR)-2-Isopropoxy-2-oxidodihydro-4H,6H-spiro[furo[3,2-d][1,3,2]dioxaphosphinine-7,2′-oxetan]-6-yl)pyrimidine-2,4(1H,3H)-dione (JNJ-54257099), a 3′-5′-Cyclic Phosphate Ester Prodrug of 2′-Deoxy-2′-Spirooxetane Uridine Triphosphate Useful for HCV Inhibition

et al. 2016

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JNJ-54257099 (9) is a novel cyclic phosphate ester derivative that belongs to the class of 2'-deoxy-2'-spirooxetane uridine nucleotide prodrugs which are known as inhibitors of the HCV NS5B RNA-dependent RNA polymerase (RdRp). In the Huh-7 HCV genotype (GT) 1b replicon-containing cell line 9 is devoid of any anti-HCV activity, an observation attributable to inefficient prodrug metabolism which was found to be CYP3A4-dependent. In contrast, in vitro incubation of 9 in primary human hepatocytes as well as pharmacokinetic evaluation thereof in different preclinical species reveals the formation of substantial levels of 2'-deoxy-2'-spirooxetane uridine triphosphate (8), a potent inhibitor of the HCV NS5B polymerase. Overall, it was found that 9 displays a superior profile compared to its phosphoramidate prodrug analogues (e.g., 4) described previously. Of particular interest is the in vivo dose dependent reduction of HCV RNA observed in HCV infected (GT1a and GT3a) human hepatocyte chimeric mice after 7 days of oral administration of 9.

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2′-Deoxy-2′-spirocyclopropylcytidine Revisited: A New and Selective Inhibitor of the Hepatitis C Virus NS5B Polymerase

Cited by 54 publications

References 56 publications

Antiviral Activity and Mode of Action of TMC647078, a Novel Nucleoside Inhibitor of the Hepatitis C Virus NS5B Polymerase

Antiviral Activity and Mode of Action of TMC647078, a Novel Nucleoside Inhibitor of the Hepatitis C Virus NS5B Polymerase

Discovery of 2′-α-C-Methyl-2′-β-C-fluorouridine Phosphoramidate Prodrugs as Inhibitors of Hepatitis C Virus

Discovery of 1-((2R,4aR,6R,7R,7aR)-2-Isopropoxy-2-oxidodihydro-4H,6H-spiro[furo[3,2-d][1,3,2]dioxaphosphinine-7,2′-oxetan]-6-yl)pyrimidine-2,4(1H,3H)-dione (JNJ-54257099), a 3′-5′-Cyclic Phosphate Ester Prodrug of 2′-Deoxy-2′-Spirooxetane Uridine Triphosphate Useful for HCV Inhibition

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