Specific Targeting of Highly Conserved Residues in the HIV-1 Reverse Transcriptase Primer Grip Region. 2. Stereoselective Interaction to Overcome the Effects of Drug Resistant Mutations

Butini, Stefania; Brindisi, Margherita; Cosconati, Sandro; Marinelli, Luciana; Borrelli, Giuseppe; Coccone, Salvatore Sanna; Ramunno, Anna; Campiani, Giuseppe; Novellino, Ettore; Zanoli, Samantha; Samuele, Alberta; Giorgi, Gianluca; Bergamini, Alberto; Mattia, Michela Di; Lalli, Silvana; Galletti, Bruno; Gemma, Sandra; Maga, Giovanni

doi:10.1021/jm801395v

Cited by 15 publications

(8 citation statements)

References 12 publications

Supporting

Mentioning

Contrasting

Unclassified

Order By: Relevance

“…The observation that mutations in the NNRTI BP that increased EFV and ETR resistance were sensitive to the resistance-enhancing effect of RNase H cleavage activity is consistent with this hypothesis. The NNRTI BP mutations analyzed in our studies were previously shown to reduce the affinity of NVP and EFV to the RT and RT-T/P (8,31,47,53,61). Although there is no biochemical data available at this time, it is likely that NNRTI BP mutations that confer resistance to DLV and ETR also reduce the affinity of these drugs to the mutant RTs (10,16).…”

Section: Discussionmentioning

confidence: 84%

“…To test the hypothesis that the affinity of the NNRTI to the RT is a key component in determining whether reducing RNase H activity affects NNRTI resistance, we created NNRTI-resistant RT mutants with amino acid substitutions L100I, K103N, V106A, V179F, and Y181C in the NNRTI BP that alter the binding characteristics of the mutant RTs for the NNRTIs (8,31,47,53,61). The effects of most of these NNRTI resistance mutations, as well as other mutations, on NVP and EFV affinity to the RT are summarized in Table 2.…”

Section: Affinity Of Nnrtis To the Rt Is A Critical Parameter In Detementioning

confidence: 99%

See 1 more Smart Citation

A Novel Molecular Mechanism of Dual Resistance to Nucleoside and Nonnucleoside Reverse Transcriptase Inhibitors

Nikolenko

Delviks-Frankenberry

Pathak

2010

J Virol

View full text Add to dashboard Cite

Recently, mutations in the connection subdomain (CN) and RNase H domain of HIV-1 reverse transcriptase (RT) were observed to exhibit dual resistance to nucleoside and nonnucleoside reverse transcriptase inhibitors (NRTIs and NNRTIs). To elucidate the mechanism by which CN and RH mutations confer resistance to NNRTIs, we hypothesized that these mutations reduce RNase H cleavage and provide more time for the NNRTI to dissociate from the RT, resulting in the resumption of DNA synthesis and enhanced NNRTI resistance. We observed that the effect of the reduction in RNase H cleavage on NNRTI resistance is dependent upon the affinity of each NNRTI to the RT and further influenced by the presence of NNRTI-binding pocket (BP) mutants. D549N, Q475A, and Y501A mutants, which reduce RNase H cleavage, enhance resistance to nevirapine (NVP) and delavirdine (DLV), but not to efavirenz (EFV) and etravirine (ETR), consistent with their increase in affinity for RT. Combining the D549N mutant with NNRTI BP mutants further increases NNRTI resistance from 3- to 30-fold, supporting the role of NNRTI-RT affinity in our NNRTI resistance model. We also demonstrated that CNs from treatment-experienced patients, previously reported to enhance NRTI resistance, also reduce RNase H cleavage and enhance NNRTI resistance in the context of the patient RT pol domain or a wild-type pol domain. Together, these results confirm key predictions of our NNRTI resistance model and provide support for a unifying mechanism by which CN and RH mutations can exhibit dual NRTI and NNRTI resistance.

show abstract

Section: Discussionmentioning

confidence: 84%

Section: Affinity Of Nnrtis To the Rt Is A Critical Parameter In Detementioning

confidence: 99%

A Novel Molecular Mechanism of Dual Resistance to Nucleoside and Nonnucleoside Reverse Transcriptase Inhibitors

Nikolenko

Delviks-Frankenberry

Pathak

2010

J Virol

View full text Add to dashboard Cite

show abstract

“…Given that most NNRTIs against HIV-1 RT are not active against HIV-2 RT (37), it is unlikely that the already established anti-HIV-1 drugs will be active against the more distantly related HBV P protein. However, by the screening of compound libraries that maintain the principal HIV-1 NNRTI butterfly-like shape (13,35) and by advanced modeling, NNRTIs with activity against HBV should be identifiable. Because all current anti-HBV therapies beyond type I interferon rely exclusively on nucleos(t)ide analogs, such NNRTIs, with their different mechanism of action, would be a most desired alternative for, or complement to, the limited drug repertoire available against chronic hepatitis B.…”

Section: Discussionmentioning

confidence: 99%

“…Together with HIV-1 RT homology-based modeling and the functionality of chimeric P proteins carrying box E, or even the sequence encompassing the catalytic YMDD motif in box C to box E, from HIV-1 RT, these data demonstrate a common catalytic core architecture of the RT domains of hepadnaviral P proteins and retroviral RTs, in particular the formation of a primer grip-like structure by the box E residues in P proteins. Because the primer grip in HIV-1 RT is the target for multiple approved and experimental NNRTIs (13,18), the presence of a primer grip equivalent in P proteins suggests that related compounds might be developed into a new class of HBV inhibitors.…”

Section: Figmentioning

confidence: 99%

Extensive Mutagenesis of the Conserved Box E Motif in Duck Hepatitis B Virus P Protein Reveals Multiple Functions in Replication and a Common Structure with the Primer Grip in HIV-1 Reverse Transcriptase

Wang

Luo

Zhao

et al. 2012

J Virol

View full text Add to dashboard Cite

c Hepadnaviruses, including the pathogenic hepatitis B virus (HBV), replicate their small DNA genomes through protein-primed reverse transcription, mediated by the terminal protein (TP) domain in their P proteins and an RNA stem-loop, ⑀, on the pregenomic RNA (pgRNA). No direct structural data are available for P proteins, but their reverse transcriptase (RT) domains contain motifs that are conserved in all RTs (box A to box G), implying a similar architecture; however, experimental support for this notion is limited. Exploiting assays available for duck HBV (DHBV) but not the HBV P protein, we assessed the functional consequences of numerous mutations in box E, which forms the DNA primer grip in human immunodeficiency virus type 1 (HIV-1) RT. This substructure coordinates primer 3=-end positioning and RT subdomain movements during the polymerization cycle and is a prime target for nonnucleosidic RT inhibitors (NNRTIs) of HIV-1 RT. Box E was indeed critical for DHBV replication, with the mutations affecting the folding, ⑀ RNA interactions, and polymerase activity of the P protein in a position-and amino acid side chain-dependent fashion similar to that of HIV-1 RT. Structural similarity to HIV-1 RT was underlined by molecular modeling and was confirmed by the replication activity of chimeric P proteins carrying box E, or even box C to box E, from HIV-1 RT. Hence, box E in the DHBV P protein and likely the HBV P protein forms a primer grip-like structure that may provide a new target for anti-HBV NNRTIs. Hepadnaviruses are small hepatotropic DNA viruses that infect humans and select mammals and birds. Hepatitis B virus (HBV), one of the most relevant viral pathogens of humans (20), is their prototypic member. All hepadnaviruses replicate their ϳ3.0-kb genomes by chaperone-assisted protein-primed reverse transcription (10), executed by their P proteins. These are unusual reverse transcriptases (RTs), which, beyond the common RNA-dependent and DNA-dependent DNA polymerase and RNase H (RH) domains, contain a unique terminal protein (TP) domain at their N termini (Fig. 1A). To initiate reverse transcription, the phenolic OH group of a specific Tyr residue in TP fills the role that conventionally is taken by the 3=-hydroxyl end of a nucleic acid primer (30).P proteins are translated from a greater-than-genome-length transcript, the pregenomic RNA (pgRNA), which also acts as mRNA for the viral core protein. The interaction of the P protein with an RNA stem-loop, ⑀, on the pgRNA is crucial for viral replication; it triggers the coencapsidation of pgRNA and the P protein into newly forming nucleocapsids and the synthesis of a short DNA oligonucleotide, which is templated by the bulge in ⑀ and, via its 5=-terminal nucleotide, becomes covalently attached to the Tyr residue in TP ("protein priming"). Upon transfer to a 3=-proximal acceptor site on pgRNA, the oligonucleotide is extended into fulllength minus-strand DNA, and the pgRNA template is concurrently degraded by the P protein's RH activity. Some 15 to 18 residues fro...

show abstract

“…Pyrrolo-1,5-benzoxazepines (PBOXs) and pyrrolonaphthoxazepines (PNOXs) are pharmaceutically relevant heterocycles. As a part of our research activity, a major focus was directed toward the development of novel PBOXs and PNOXs as potent antitumor agents, inhibitors of adenosine kinase and nonnucleoside inhibitors of human immunodeficiency virus reverse transcriptase (HIV-1 RT) [1][2][3][4][5][6][7][8][9][10][11][12]. In particular, our continuing efforts in the field of antitumor agents led to the development of PBOX-and PNOXbased compounds as potent proapoptotic agents [4].…”

mentioning

confidence: 99%

A Jocic-type approach for a practical and scalable synthesis of pyrrolonaphthoxazepine (PNOX)-based potent proapoptotic agents

Federico

Khan

Relitti

et al. 2018

Tetrahedron Letters

Self Cite

View full text Add to dashboard Cite

We developed a Jocic-type protocol for the construction of the pyrrolonaphthoxazepine (PNOX) core. After an initial investigation based on the isolation of a trichloromethyl carbinol derivative, we shifted our attention towards a multicomponent single-step protocol. Screening of a variety of bases and solvents led to the identification of the optimum conditions for the preparation of the key a-aryloxy carboxylic acids to undergo intramolecular cyclization. The novel chemical route significantly improved overall yields for the preparation of PNOX-based compounds and was successfully extended to the preparation of 1,4-benzoxazinone-based templates.

show abstract

Specific Targeting of Highly Conserved Residues in the HIV-1 Reverse Transcriptase Primer Grip Region. 2. Stereoselective Interaction to Overcome the Effects of Drug Resistant Mutations

Cited by 15 publications

References 12 publications

A Novel Molecular Mechanism of Dual Resistance to Nucleoside and Nonnucleoside Reverse Transcriptase Inhibitors

A Novel Molecular Mechanism of Dual Resistance to Nucleoside and Nonnucleoside Reverse Transcriptase Inhibitors

Extensive Mutagenesis of the Conserved Box E Motif in Duck Hepatitis B Virus P Protein Reveals Multiple Functions in Replication and a Common Structure with the Primer Grip in HIV-1 Reverse Transcriptase

A Jocic-type approach for a practical and scalable synthesis of pyrrolonaphthoxazepine (PNOX)-based potent proapoptotic agents

Contact Info

Product

Resources

About