The Application of Phosphoramidate Protide Technology to Acyclovir Confers Anti-HIV Inhibition

Derudas, Marco; Carta, Davide; Brancale, Andrea; Vanpouille, Christophe; Lisco, Andrea; Margolis, Leonid; Balzarini, Jan; McGuigan, Christopher

doi:10.1021/jm9007856

Cited by 72 publications

(89 citation statements)

References 16 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…The stereo-selective hydrolysis of an acyclovir phosphoramidate prodrug was previously demonstrated by using carboxypeptidase Y, a structural homolog of cathepsin A (31,32). A computer model of carboxypeptidase Y docked with a phosphoramidate prodrug in the active site suggested that positioning of the carbonyl moiety of the carboxyl ester with the R-phosphate diastereoisomer was more preferable for catalysis than with the S-diastereoisomer (31). However, they failed to identify which diastereoisomer was preferentially hydrolyzed by carboxypeptidase Y in their biochemical assay.…”

Section: Discussionmentioning

confidence: 99%

Mechanism of Activation of PSI-7851 and Its Diastereoisomer PSI-7977

Murakami¹,

Tolstykh²,

Bao³

et al. 2010

Journal of Biological Chemistry

268

308

View full text Add to dashboard Cite

A phosphoramidate prodrug of 2-deoxy-2-␣-fluoro-␤-Cmethyluridine-5-monophosphate, PSI-7851, demonstrates potent anti-hepatitis C virus (HCV) activity both in vitro and in vivo. PSI-7851 is a mixture of two diastereoisomers, PSI-7976 and PSI-7977, with PSI-7977 being the more active inhibitor of HCV RNA replication in the HCV replicon assay. To inhibit the HCV NS5B RNA-dependent RNA polymerase, PSI-7851 must be metabolized to the active triphosphate form. The first step, hydrolysis of the carboxyl ester by human cathepsin A (CatA) and/or carboxylesterase 1 (CES1), is a stereospecific reaction. Western blot analysis showed that CatA and CES1 are both expressed in primary human hepatocytes. However, expression of CES1 is undetectable in clone A replicon cells. Studies with inhibitors of CatA and/or CES1 indicated that CatA is primarily responsible for hydrolysis of the carboxyl ester in clone A cells, although in primary human hepatocytes, both CatA and CES1 contribute to the hydrolysis. Hydrolysis of the ester is followed by a putative nucleophilic attack on the phosphorus by the carboxyl group resulting in the spontaneous elimination of phenol and the production of an alaninyl phosphate metabolite, PSI-352707, which is common to both isomers. The removal of the amino acid moiety of PSI-352707 is catalyzed by histidine triad nucleotide-binding protein 1 (Hint1) to give the 5-monophosphate form, PSI-7411. siRNA-mediated Hint1 knockdown studies further indicate that Hint1 is, at least in part, responsible for converting PSI-352707 to PSI-7411. PSI-7411 is then consecutively phosphorylated to the diphosphate, PSI-7410, and to the active triphosphate metabolite, PSI-7409, by UMP-CMP kinase and nucleoside diphosphate kinase, respectively.Nucleoside analogs have long been the backbone therapy for the treatment of viral diseases such as HIV, HBV, and HSV infections (1-5). Recent studies have suggested that nucleoside analogs may be useful for treating hepatitis C virus (HCV) 3 infection (4, 6 -8). The most advanced anti-HCV nucleoside, RG7128, is a diisobutyrate nucleoside prodrug of ␤-D-2Ј-deoxy-2Ј-␣-fluoro-2Ј-␤-C-methylcytidine (PSI-6130) and is currently in phase IIb clinical studies. PSI-6130 demonstrated potent activity in the subgenomic HCV replicon assay (9); the incubation of radiolabeled PSI-6130 with either replicon cells or primary human hepatocytes resulted in the formation of the 5Ј-mono-, di-, and triphosphate metabolites of . The triphosphate metabolite (PSI-6130-TP) was shown to be a potent inhibitor of HCV NS5B RNA-directed RNA polymerase (RdRp) (11). However, incubation of replicon cells with the uridine analog, PSI-6206, resulted in no inhibition of HCV RNA production due to the inability of PSI-6206 to be phosphorylated by cellular nucleoside kinases to its monophosphate, 12). Biochemical studies showed that PSI-7411 was consecutively phosphorylated to its diphosphate, PSI-7410, by UMP-CMP kinase and its triphosphate, PSI-7409, by nucleoside diphosphate kinase (12). Inhibition studies using the replic...

show abstract

Section: Discussionmentioning

confidence: 99%

Mechanism of Activation of PSI-7851 and Its Diastereoisomer PSI-7977

Murakami¹,

Tolstykh²,

Bao³

et al. 2010

Journal of Biological Chemistry

268

308

View full text Add to dashboard Cite

show abstract

“…These compounds showed anti-HIV activity in the absence of HHV infection, demonstrating their nucleoside kinase independence. 4, 5 These findings were also supported in a different study, where ACV phosphate prodrugs showed a full retention of antiviral activity against HSV-1 and VZV thymidine kinase (TK)-deficient strains. 6 Unfortunately, these compounds proved to be somewhat cytotoxic in some assays.…”

Section: Introductionmentioning

confidence: 58%

Virtual Screening of Acyclovir Derivatives as Potential Antiviral Agents: Design, Synthesis, and Biological Evaluation of New Acyclic Nucleoside ProTides

et al. 2017

Self Cite

View full text Add to dashboard Cite

Following our findings on the anti-HIV activity of acyclovir phosphate prodrugs, we herein report the ProTide approach applied to a series of acyclic nucleosides aimed at the identification of novel and selective antiviral (i.e. HIV) agents. Acyclic nucleoside analogues used in this study were identified through a virtual screening using adenylate/guanylate kinase, HIV RT, and human DNA polymerase. A total of thirty-nine new phosphate prodrugs were synthesised and evaluated against HIV-1 (in in vitro and in ex-vivo human tonsillar tissue system) and human herpesviruses. Several ProTide compounds showed good potency (low micromolar range) against HIV-1 while the parent nucleosides were not effective. Also, pronounced inhibition of herpesvirus replication was observed. A carboxypeptidase-mediated hydrolysis study was performed for the selection of compounds showing different metabolic patterns.

show abstract

“…The final coupling of the nucleoside 25 has been performed using an excess of the phosphorochloridate (33) (3 eq) in the presence of t BuMgCl (3eq) following the procedure reported by Uchiyama [17] and extensively used for the synthesis of the ProTides [18]. The desired compound 34 was obtained as a mixture of two diastereoisomers confirmed by the presence of two peaks in the 31 P NMR (2.92, 2.27).…”

Section: Chemical Synthesismentioning

confidence: 99%

Synthesis and antiviral evaluation of α-l-2′-deoxythreofuranosyl nucleosides

Toti

Derudas

McGuigan

et al. 2011

European Journal of Medicinal Chemistry

Self Cite

View full text Add to dashboard Cite

Th e syn t h e sis o f a series o f α-L-2'-deoxythreofuranosyl nucleosides featuring the nucleobases A, T, C and U is described in seven steps from 1,2-O-isopropyledene-α-Lthreose, involving a Vorbrüggen coupling and a Barton-McCombie deoxygenation protocol as the key steps. All analogues, including a phosphoramidate nucleoside phosphate prodrug of the T analogue, were evaluated against a broad panel of different viruses but found inactive, while also lacking notable cellular toxicity. The thymidine analogue showed inhibition to mitochondrial thymidine kinase-2 (TK-2), herpes simplex virus type 1 (HSV-1) TK, varicella-zoster virus (VZV) TK and M. tuberculosis thymidylate kinase.

show abstract

The Application of Phosphoramidate Protide Technology to Acyclovir Confers Anti-HIV Inhibition

Cited by 72 publications

References 16 publications

Mechanism of Activation of PSI-7851 and Its Diastereoisomer PSI-7977

Mechanism of Activation of PSI-7851 and Its Diastereoisomer PSI-7977

Virtual Screening of Acyclovir Derivatives as Potential Antiviral Agents: Design, Synthesis, and Biological Evaluation of New Acyclic Nucleoside ProTides

Synthesis and antiviral evaluation of α-l-2′-deoxythreofuranosyl nucleosides

Contact Info

Product

Resources

About