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

Abstract

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.

Tissue-Specific Proteomics Analysis of Anti-COVID-19 Nucleoside and Nucleotide Prodrug-Activating Enzymes Provides Insights into the Optimization of Prodrug Design and Pharmacotherapy Strategy

“…Hydrolysis efficiency of two cyclic monophosphate prodrugs. (a) Putative the activation pathway of PSI-352938 and JNJ-54257099. , (b) Protein expression of CYP3A4 in different human tissues. Data are the means of three independent measurements with error bars representing SD.…”

“…Both prodrugs were originally developed as orally dosed anti-HCV drugs. Their activations were initiated by the removal of the isopropyl group in the cyclic phosphate moiety, which was mainly catalyzed by cytochrome P450 3A4 (CYP3A4) (Figure a). , The activities of CYP3A4 on PSI-352938 and JNJ-54257099 were calculated to be 2.27 and 1.32 pmol/min/pmol of CYP3A4, respectively, based on the intrinsic clearance reported in a previous study . The original study of PSI-352938 tested 16 hydrolases (e.g., CESs, cathepsins, and lipase) and multiple CYPs, and it identified CYP3A4 as the only enzyme involved in the initial activation of the prodrug .…”

“…GS-465124, GS-6620, and GS2 were included because of their structural similarity to remdesivir: In the nucleoside moiety, these compounds differ from remdesivir only by a methyl group on the 2′ pentose ring. PSI-352938 and JNJ-54257099 are cyclic phosphate prodrugs. Valaciclovir and 5′-O- l -valyl-decitabine ( l -val-DAC) were selected as examples of l -valyl ester prodrugs .…”

“…Laninamivir octanoate was selected as an example of a long-acting inhaled antiviral prodrug . The activating enzymes and activation mechanisms of these prodrugs were obtained from the literature and carefully reviewed and scrutinized. ,,− …”

Phosphoramidates and phosphonamidates (ProTides) with antiviral activity

“…Currently, several academic groups and pharmaceutical companies have more ProTide compounds in the pipeline undergoing preclinical studies for the treatment of viral infections. 55,179–185…”