Specific Targeting Highly Conserved Residues in the HIV-1 Reverse Transcriptase Primer Grip Region. Design, Synthesis, and Biological Evaluation of Novel, Potent, and Broad Spectrum NNRTIs with Antiviral Activity

Fattorusso, Caterina; Gemma, Sandra; Butini, Stefania; Huleatt, Paul B.; Catalanotti, Bruno; Persico, Marco; Angelis, Meri De; Fiorini, Isabella; Nacci, Vito; Ramunno, Anna; Rodriquez, Manuela; Greco, Giovanni; Novellino, Ettore; Bergamini, Alberto; Marini, Stefano; Coletta, Massimo; Maga, Giovanni; Spadari, Silvio; Campiani, Giuseppe

doi:10.1021/jm050257d

Cited by 38 publications

(27 citation statements)

References 34 publications

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“…This residue not only is part of the NNRTI binding pocket but also belongs to the "primer grip" of HIV RT, and any mutation at this position has been shown to severely compromise the RNA-and DNA-dependent DNA polymerase activities of the enzyme (33). Indeed, this immutable residue has become the focus of the development of novel NNRTIs (17). In addition, upon the binding of many NNRTIs, the side chain of Tyr181, contained within the ␤4-␤7-␤8 sheet, have been reported to flip conformation from a "down" to an "up" position (38).…”

Section: Discussionmentioning

confidence: 99%

Lersivirine, a Nonnucleoside Reverse Transcriptase Inhibitor with Activity against Drug-Resistant Human Immunodeficiency Virus Type 1

Corbau¹,

Mori²,

Phillips

et al. 2010

Antimicrob Agents Chemother

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The nonnucleoside reverse transcriptase inhibitors (NNRTIs) are key components of highly active antiretroviral therapy (HAART) for the treatment of human immunodeficiency virus type 1 (HIV-1). A major problem with the first approved NNRTIs was the emergence of mutations in the HIV-1 reverse transcriptase (RT), in particular K103N and Y181C, which led to resistance to the entire class. We adopted an iterative strategy to synthesize and test small molecule inhibitors from a chemical series of pyrazoles against wild-type (wt) RT and the most prevalent NNRTI-resistant mutants. The emerging candidate, lersivirine (UK-453,061), binds the RT enzyme in a novel way (resulting in a unique resistance profile), inhibits over 60% of viruses bearing key RT mutations, with 50% effective concentrations (EC 50 s) within 10-fold of those for wt viruses, and has excellent selectivity against a range of human targets. Altogether lersivirine is a highly potent and selective NNRTI, with excellent efficacy against NNRTI-resistant viruses.

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Section: Discussionmentioning

confidence: 99%

Lersivirine, a Nonnucleoside Reverse Transcriptase Inhibitor with Activity against Drug-Resistant Human Immunodeficiency Virus Type 1

Corbau¹,

Mori²,

Phillips

et al. 2010

Antimicrob Agents Chemother

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“…It is interesting that TMC125, a second generation NNRTI, efficiently inhibits HIV replication in M/M, even if at a concentration similar to those observed in CD4ϩ T lymphocytes [64]. In particular, it has been shown that the EC 50 of TMC125 against the laboratory strain HIV Bal is in the lower nanomolar range [2.0 (0.8 -4.0)] nm and highly similar to that observed for efavirenz {2.0 (0.9 -3.0) nm [64]}. The HIV Bal replication in M/M is also inhibited efficiently by the pyrrolobenzoxazepinones, a new class of NNRTIs designed to target the highly conserved primer grip within the ␣12-␣13 hairpin.…”

Section: Nnrtismentioning

confidence: 97%

“…Indeed, once the proviral DNA is integrated into the host genome, the production of viral particles is independent of the RT enzyme and thus, is not affected by the RTIs. For these reasons, the activity of several drugs acting at a late stage of HIV replication (anti-rev, anti-tat, transcription inhibitors, IFN-␣, IFN-␥, ampligen) has been tested in chronically infected M/M [64]. However, results were not encouraging, as all these drugs failed to suppress HIV replication.…”

Section: Protease Inhibitorsmentioning

confidence: 99%

Mechanisms underlying activity of antiretroviral drugs in HIV-1-infected macrophages: new therapeutic strategies

Aquaro

Svicher

Schols

et al. 2006

Journal of Leukocyte Biology

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Monocyte-derived macrophages (M/M) are considered the second cellular target of HIV-1 and a crucial virus reservoir. M/M are widely distributed in all tissues and organs, including the CNS, where they represent the most common HIV-infected cells. Differently from activated CD4+ T lymphocytes, M/M are resistant to the cytopathic effect of HIV and survive HIV infection for a long time. Moreover, HIV-1 replication in M/M is a key pathogenetic event during the course of HIV-1 infection. Overall findings strongly support the clinical relevance of anti-HIV drugs in M/M. Nucleoside RT inhibitors (NRTIs) are more active against HIV in M/M than in CD4+ T lymphocytes. Their activity is further boosted by the presence of an additional monophosphate group (i.e., a phosphonate group, as in the case of Tenofovir), thus overcoming the bottleneck of the low phosphorylation ability of M/M. In contrast, the antiviral activity of non-NRTIs (not affecting the DNA chain elongation) in M/M is similar to that in CD4+ T lymphocytes. Protease inhibitors are the only clinically approved drugs acting at a late stage of the HIV lifecycle. They are able to interfere with HIV replication in HIV-1 chronically infected M/M, even if at concentrations greater than those observed in HIV-1 chronically infected CD4+ T lymphocytes. Finally, several new drugs have been shown to interfere efficiently with HIV replication in M/M, including entry inhibitors. A better understanding of the activity of the anti-HIV drugs in M/M may represent a key element for the design of effective anti-HIV chemotherapy.

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“…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

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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...

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Specific Targeting Highly Conserved Residues in the HIV-1 Reverse Transcriptase Primer Grip Region. Design, Synthesis, and Biological Evaluation of Novel, Potent, and Broad Spectrum NNRTIs with Antiviral Activity

Cited by 38 publications

References 34 publications

Lersivirine, a Nonnucleoside Reverse Transcriptase Inhibitor with Activity against Drug-Resistant Human Immunodeficiency Virus Type 1

Lersivirine, a Nonnucleoside Reverse Transcriptase Inhibitor with Activity against Drug-Resistant Human Immunodeficiency Virus Type 1

Mechanisms underlying activity of antiretroviral drugs in HIV-1-infected macrophages: new therapeutic strategies

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

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