Potent competitive inhibition of human ribonucleotide reductase by a nonnucleoside small molecule

Ahmad, Md. Faiz; Alam, Intekhab; Huff, Sarah E.; Pink, John; Flanagan, Sheryl A.; Shewach, Donna; Misko, Tessianna A.; Oleinick, Nancy L.; Harte, William E.; Viswanathan, Rajesh; Harris, Michael E.; Dealwis, Chris Godfrey

doi:10.1073/pnas.1620220114

Title: Potent competitive inhibition of human ribonucleotide reductase by a nonnucleoside small molecule

Journal Article · Mon Jul 17 00:00:00 EDT 2017 · Proceedings of the National Academy of Sciences of the United States of America

DOI:https://doi.org/10.1073/pnas.1620220114· OSTI ID:1371602

Ahmad, Md. Faiz; Alam, Intekhab; Huff, Sarah E.; Pink, John; Flanagan, Sheryl A.; Shewach, Donna; Misko, Tessianna A.; Oleinick, Nancy L.; Harte, William E.; Viswanathan, Rajesh; Harris, Michael E.;

Human ribonucleotide reductase (hRR) is crucial for DNA replication and maintenance of a balanced dNTP pool, and is an established cancer target. Nucleoside analogs such as gemcitabine diphosphate and clofarabine nucleotides target the large subunit (hRRM1) of hRR. These drugs have a poor therapeutic index due to toxicity caused by additional effects, including DNA chain termination. The discovery of nonnucleoside, reversible, small-molecule inhibitors with greater specificity against hRRM1 is a key step in the development of more effective treatments for cancer. Furthermore, we report the identification and characterization of a unique nonnucleoside small-molecule hRR inhibitor, naphthyl salicylic acyl hydrazone (NSAH), using virtual screening, binding affinity, inhibition, and cell toxicity assays. NSAH binds to hRRM1 with an apparent dissociation constant of 37 µM, and steady-state kinetics reveal a competitive mode of inhibition. A 2.66-Å resolution crystal structure of NSAH in complex with hRRM1 demonstrates that NSAH functions by binding at the catalytic site (C-site) where it makes both common and unique contacts with the enzyme compared with NDP substrates. Importantly, the IC₅₀for NSAH is within twofold of gemcitabine for growth inhibition of multiple cancer cell lines, while demonstrating little cytotoxicity against normal mobilized peripheral blood progenitor cells. NSAH depresses dGTP and dATP levels in the dNTP pool causing S-phase arrest, providing evidence for RR inhibition in cells. This report of a nonnucleoside reversible inhibitor binding at the catalytic site of hRRM1 provides a starting point for the design of a unique class of hRR inhibitors.

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Research Organization:: Argonne National Laboratory (ANL), Argonne, IL (United States)

Sponsoring Organization:: USDOE Office of Science (SC), Basic Energy Sciences (BES). Scientific User Facilities Division; National Institutes of Health (NIH); National Inst. of General Medical Sciences

Grant/Contract Number:: AC02-06CH11357; AC02-05CH11231; R01GM100887; P30 CA43703; P41 GM103403

OSTI ID:: 1371602

Alternate ID(s):: OSTI ID: 1390861

Journal Information:: Proceedings of the National Academy of Sciences of the United States of America, Journal Name: Proceedings of the National Academy of Sciences of the United States of America Vol. 114 Journal Issue: 31; ISSN 0027-8424

Publisher:: National Academy of SciencesCopyright Statement

Country of Publication:: United States

Language:: English

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Cited by: 20 works

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