Unknown

Dataset Information

0

Identification of mechanistically distinct inhibitors of HIV-1 reverse transcriptase through fragment screening.


ABSTRACT: Fragment-based screening methods can be used to discover novel active site or allosteric inhibitors for therapeutic intervention. Using saturation transfer difference (STD) NMR and in vitro activity assays, we have identified fragment-sized inhibitors of HIV-1 reverse transcriptase (RT) with distinct chemical scaffolds and mechanisms compared to nonnucleoside RT inhibitors (NNRTIs) and nucleoside/nucleotide RT inhibitors (NRTIs). Three compounds were found to inhibit RNA- and DNA-dependent DNA polymerase activity of HIV-1 RT in the micromolar range while retaining potency against RT variants carrying one of three major NNRTI resistance mutations: K103N, Y181C, or G190A. These compounds also inhibit Moloney murine leukemia virus RT but not the Klenow fragment of Escherichia coli DNA polymerase I. Steady-state kinetic analyses demonstrate that one of these fragments is a competitive inhibitor of HIV-1 RT with respect to deoxyribonucleoside triphosphate (dNTP) substrate, whereas a second compound is a competitive inhibitor of RT polymerase activity with respect to the DNA template/primer (T/P), and consequently also inhibits RNase H activity. The dNTP competing RT inhibitor retains activity against the NRTI-resistant mutants K65R and M184V, demonstrating a drug resistance profile distinct from the nucleotide competing RT inhibitors indolopyridone-1 (INDOPY-1) and 4-dimethylamino-6-vinylpyrimidine-1 (DAVP-1). In antiviral assays, the T/P competing compound inhibits HIV-1 replication at a step consistent with an RT inhibitor. Screening of additional structurally related compounds to the three fragments led to the discovery of molecules with improved potency against HIV-1 RT. These fragment inhibitors represent previously unidentified scaffolds for development of novel drugs for HIV-1 prevention or treatment.

SUBMITTER: La J 

PROVIDER: S-EPMC4460473 | biostudies-literature | 2015 Jun

REPOSITORIES: biostudies-literature

altmetric image

Publications

Identification of mechanistically distinct inhibitors of HIV-1 reverse transcriptase through fragment screening.

La Jennifer J   Latham Catherine F CF   Tinetti Ricky N RN   Johnson Adam A   Tyssen David D   Huber Kelly D KD   Sluis-Cremer Nicolas N   Simpson Jamie S JS   Headey Stephen J SJ   Chalmers David K DK   Tachedjian Gilda G  

Proceedings of the National Academy of Sciences of the United States of America 20150518 22


Fragment-based screening methods can be used to discover novel active site or allosteric inhibitors for therapeutic intervention. Using saturation transfer difference (STD) NMR and in vitro activity assays, we have identified fragment-sized inhibitors of HIV-1 reverse transcriptase (RT) with distinct chemical scaffolds and mechanisms compared to nonnucleoside RT inhibitors (NNRTIs) and nucleoside/nucleotide RT inhibitors (NRTIs). Three compounds were found to inhibit RNA- and DNA-dependent DNA p  ...[more]

Similar Datasets

| S-EPMC3906421 | biostudies-literature
| S-EPMC3999242 | biostudies-literature
| S-EPMC2904523 | biostudies-literature
| S-EPMC3144253 | biostudies-literature
| S-EPMC6158299 | biostudies-literature
2018-12-12 | E-MTAB-7087 | biostudies-arrayexpress
| S-EPMC2573056 | biostudies-literature
| S-EPMC9279714 | biostudies-literature
| S-EPMC5046171 | biostudies-literature
| S-EPMC8654897 | biostudies-literature