scholarly journals Selectivity for strand-transfer over 3′-processing and susceptibility to clinical resistance of HIV-1 integrase inhibitors are driven by key enzyme–DNA interactions in the active site

2016 ◽  
Vol 44 (14) ◽  
pp. 6896-6906 ◽  
Author(s):  
Mathieu Métifiot ◽  
Barry C. Johnson ◽  
Evgeny Kiselev ◽  
Laura Marler ◽  
Xue Zhi Zhao ◽  
...  
Keyword(s):  
Active Site ◽  
Strand Transfer ◽  
Integrase Inhibitors ◽  
Dna Interactions ◽  
Clinical Resistance ◽  
Key Enzyme ◽  
Hiv 1

scholarly journals Characterization of HIV-1 Integrase Gene and Resistance Associated Mutations Prior to Roll out of Integrase Inhibitors by Kenyan National HIV-Treatment Program in Kenya

2020 ◽  
Vol 30 (1) ◽  
Author(s):  
Mabeya Sepha ◽  
Nyamache Anthony ◽  
Ngugi Caroline ◽  
Nyerere Andrew ◽  
Lihana Raphael
Keyword(s):  
Drug Resistance ◽  
Direct Sequencing ◽  
Integrase Inhibitor ◽  
Hiv Treatment ◽  
Strand Transfer ◽  
Hiv Integrase ◽  
Integrase Inhibitors ◽  
Resistance Mutations ◽  
Integrase Gene ◽  
Hiv 1

BACKGROUND: Antiretroviral therapy containing an integrase strand transfer inhibitor plus two Nucleoside Reverse Transcriptase inhibitors has now been recommended for treatment of HIV-1-infected patients. This thus determined possible pre-existing integrase resistance associated mutations in the integrase gene prior to introduction of integrase inhibitors combination therapy in Kenya.METHODS: Drug experienced HIV patients were enrolled at Kisii Teaching and Referral in Kenya. Blood specimens from (33) patients were collected for direct sequencing of HIV-1 polintegrase genes. Drug resistance mutations were interpreted according to the Stanford algorithm and phylogenetically analysed using insilico tools.RESULTS: From pooled 188 Kenyan HIV integrase sequences that were analysed for drug resistance, no major mutations conferring resistance to integrase inhibitors were detected. However, polymorphic accessory mutations associated with reduced susceptibility of integrase inhibitors were observed in low frequency; M50I (12.2%), T97A (3.7%), S153YG, E92G (1.6%), G140S/A/C (1.1%) and E157Q (0.5%). Phylogenetic analysis (330 sequences revealed that HIV-1 subtype A1 accounted for majority of the infections, 26 (78.8%), followed by D, 5 (15.2%) and C, 2 (6%).CONCLUSION: The integrase inhibitors will be effective in Kenya where HIV-1 subtype A1 is still the most predominant. However, occurring polymorphisms may warrant further investigation among drug experienced individuals on dolutegravir combination or integrase inhibitor treatment. 

scholarly journals HIV-1 Integrase-Targeted Short Peptides Derived from a Viral Protein R Sequence

Molecules ◽  
2018 ◽  
Vol 23 (8) ◽  
pp. 1858 ◽  
Author(s):  
Xue Zhao ◽  
Mathieu Métifiot ◽  
Evgeny Kiselev ◽  
Jacques Kessl ◽  
Kasthuraiah Maddali ◽  
...  
Keyword(s):  
Viral Protein ◽  
Divalent Metal ◽  
Common Mechanism ◽  
Strand Transfer ◽  
Divalent Metal Ions ◽  
Dna Interactions ◽  
Inhibitory Potency ◽  
New Class ◽  
Viral Protein R ◽  
Hiv 1

HIV-1 integrase (IN) inhibitors represent a new class of highly effective anti-AIDS therapeutics. Current FDA-approved IN strand transfer inhibitors (INSTIs) share a common mechanism of action that involves chelation of catalytic divalent metal ions. However, the emergence of IN mutants having reduced sensitivity to these inhibitors underlies efforts to derive agents that antagonize IN function by alternate mechanisms. Integrase along with the 96-residue multifunctional accessory protein, viral protein R (Vpr), are both components of the HIV-1 pre-integration complex (PIC). Coordinated interactions within the PIC are important for viral replication. Herein, we report a 7-mer peptide based on the shortened Vpr (69–75) sequence containing a biotin group and a photo-reactive benzoylphenylalanyl residue, and which exhibits low micromolar IN inhibitory potency. Photo-crosslinking experiments have indicated that the peptide directly binds IN. The peptide does not interfere with IN-DNA interactions or induce higher-order, aberrant IN multimerization, suggesting a mode of action for the peptide that is distinct from clinically used INSTIs and developmental allosteric IN inhibitors. This compact Vpr-derived peptide may serve as a valuable pharmacological tool to identify a potential new pharmacologic site.

scholarly journals Screening of Non-catalytic Integrase Inhibitors for Anti-HIV Drug Development

2020 ◽  
Author(s):  
Ki Hoon Park ◽  
Minjee Kim ◽  
Seoung Eun Bae ◽  
Hee Jung Lee ◽  
Kyung-Chang Kim ◽  
...  
Keyword(s):  
Antiviral Activity ◽  
Drug Efficacy ◽  
Treatment Method ◽  
Strand Transfer ◽  
Inhibition Assay ◽  
Integrase Inhibitors ◽  
Catalytic Core ◽  
Host Chromosome ◽  
Inconsistent Result ◽  
Hiv 1

Abstract Background: Integrase (IN) is an essential protein for HIV replication that catalyzes insertion of the reverse-transcribed viral genome into the host chromosome during the early steps of viral infection. Highly active anti-retroviral therapy (HAART) is a HIV/AIDS treatment method that combines three or more antiviral drugs often formulated from compounds that inhibit the activities of viral reverse transcriptase and protease enzymes. Early IN inhibitors (INIs) mainly serve as integrase strand transfer inhibitors (INSTI) that disrupt strand transfer by binding the catalytic core domain (CCD) of IN. However, mutations of IN can confer resistance to INSTI. Therefore, non-catalytic integrase inhibitors (NCINI) have been developed as next-generation INIs. Methods: In this study, we evaluated and compared the activity of INSTI and NCINI according to the analysis method. Antiviral activity was compared using p24 ELISA with MT2 cell and TZM-bl luciferase system with TZM-bl cell. Each drug was serially diluted and treated to MT2 and TZM-b1 cells, infected with HIV-1 AD8 strain and incubated for 5 and 2 days, respectively. Additionally, to analyze properties of INSTI and NCINI, transfer inhibition assay and 3'-processing inhibition assay were performed. Results: During screening of INIs using the p24 ELISA and TZM-bl luciferase systems, we found an inconsistent result with INSTI and NCINI drugs. Following infection of MT2 and TZM-bl cells with T-tropic HIV-1 strain, both INSTI and NCINI treatments induced significant p24 reduction in MT2 cells. However, NCINI showed no antiviral activity in the TZM-bl luciferase system, indicating that this widely used and convenient antiretroviral assay is not suitable for screening of NCINI compounds that target the second round of HIV-1 replication. Conclusion: Accordingly, we recommend application of other assay procedures, such as p24 ELISA or reverse transcription activity, in lieu of the TZM-bl luciferase system for preliminary NCINI drug screening. Utilization of appropriate analytical methods based on underlying mechanisms is necessary for accurate assessment of drug efficacy.

scholarly journals Spectrometric and computational studies of the binding of HIV-1 integrase inhibitors to viral DNA extremities

2019 ◽  
Author(s):  
Lea El Khoury ◽  
Krystel El Hage ◽  
Jean-Philip Piquemal ◽  
Serge Fermandjian ◽  
Richard Maroun ◽  
...  
Keyword(s):  
Density Functional ◽  
Strand Transfer ◽  
Free Energies ◽  
Integrase Inhibitors ◽  
Resistance Mutations ◽  
Viral Dna ◽  
Functional Theory ◽  
Fluorescence Titrations ◽  
Dynamics Simulations ◽  
Hiv 1

Three Integrase (IN) strand transfer inhibitors are in intensive clinical use, raltegravir, elvitegravir anddolutegravir. However, the onset of IN resistance mutations limits their therapeutic efficiency. As put forth earlier, the drug affinity for the intasome could be improved by targeting preferentially the retroviralnucleobases, which are little, if at all, mutation-prone. We report experimental results of anisotropy fluorescence titrations of viral DNA by these three drugs . These show that the ranking of their inhibitory activities of the intasome corresponds to that of their free energies of binding, D Gs,to retroviral DNA, and that such a ranking is only governed by the binding enthalpies, D H, the entropy undergoing marginal variations.This ranking can therefore be directly correlated to that of model Quantum Chemistry (QC) calculations of intermolecular interaction energies of the sole halobenzene ring with the highly conserved retroviral nucleobases G4 and C14, using Density Functional Theory. This DE(QC) ranking is in turn reproduced by the corresponding DE tot values computed with a polarizable molecular mechanics/dynamics procedure, SIBFA (Sum of Interactions Between Fragments Ab initio computed). Such validations should enable polarizable molecular dynamics simulations on more potent inhibitors in their complexes with the complete intasome. Such derivatives should principally encompass modified halobenzene rings.

Correlation of Biological Activity with Active Site Binding Modes of Geminal Disulfone HIV-1 Integrase Inhibitors

ChemMedChem ◽  
2006 ◽  
Vol 1 (9) ◽  
pp. 959-964 ◽  
Author(s):  
D. Christopher Meadows ◽  
Dean J. Tantillo ◽  
Jacquelyn Gervay-Hague
Keyword(s):  
Biological Activity ◽  
Active Site ◽  
Binding Modes ◽  
Integrase Inhibitors ◽  
Hiv 1 ◽  
Site Binding

Active Site Binding Modes of HIV-1 Integrase Inhibitors

2000 ◽  
Vol 43 (22) ◽  
pp. 4109-4117 ◽  
Author(s):  
Christoph A. Sotriffer ◽  
Ni ◽  
J. Andrew McCammon
Keyword(s):  
Active Site ◽  
Binding Modes ◽  
Integrase Inhibitors ◽  
Hiv 1 ◽  
Site Binding

scholarly journals Developing and Evaluating Inhibitors against the RNase H Active Site of HIV-1 Reverse Transcriptase

2018 ◽  
Vol 92 (13) ◽  
Author(s):  
Paul L. Boyer ◽  
Steven J. Smith ◽  
Xue Zhi Zhao ◽  
Kalyan Das ◽  
Kevin Gruber ◽  
...  
Keyword(s):  
Reverse Transcriptase ◽  
Active Site ◽  
Nucleoside Analogs ◽  
Polymerase Activity ◽  
Rnase H ◽  
Strand Transfer ◽  
Hiv Replication ◽  
Content Type ◽  
Rnase Hi ◽  
Hiv 1

ABSTRACT We tested three compounds for their ability to inhibit the RNase H (RH) and polymerase activities of HIV-1 reverse transcriptase (RT). A high-resolution crystal structure (2.2 Å) of one of the compounds showed that it chelates the two magnesium ions at the RH active site; this prevents the RH active site from interacting with, and cleaving, the RNA strand of an RNA-DNA heteroduplex. The compounds were tested using a variety of substrates: all three compounds inhibited the polymerase-independent RH activity of HIV-1 RT. Time-of-addition experiments showed that the compounds were more potent if they were bound to RT before the nucleic acid substrate was added. The compounds significantly inhibited the site-specific cleavage required to generate the polypurine tract (PPT) RNA primer that initiates the second strand of viral DNA synthesis. The compounds also reduced the polymerase activity of RT; this ability was a result of the compounds binding to the RH active site. These compounds appear to be relatively specific; they do not inhibit either Escherichia coli RNase HI or human RNase H2. The compounds inhibit the replication of an HIV-1-based vector in a one-round assay, and their potencies were only modestly decreased by mutations that confer resistance to integrase strand transfer inhibitors (INSTIs), nucleoside analogs, or nonnucleoside RT inhibitors (NNRTIs), suggesting that their ability to block HIV replication is related to their ability to block RH cleavage. These compounds appear to be useful leads that can be used to develop more potent and specific compounds. IMPORTANCE Despite advances in HIV-1 treatment, drug resistance is still a problem. Of the four enzymatic activities found in HIV-1 proteins (protease, RT polymerase, RT RNase H, and integrase), only RNase H has no approved therapeutics directed against it. This new target could be used to design and develop new classes of inhibitors that would suppress the replication of the drug-resistant variants that have been selected by the current therapeutics.

scholarly journals HIV-1 Integrase Inhibitors That Are Broadly Effective against Drug-Resistant Mutants

2018 ◽  
Vol 62 (9) ◽  
Author(s):  
Steven J. Smith ◽  
Xue Zhi Zhao ◽  
Terrence R. Burke ◽  
Stephen H. Hughes
Keyword(s):  
Transfer Reaction ◽  
Drug Resistant ◽  
Strand Transfer ◽  
Integrase Inhibitors ◽  
Integrase Strand Transfer Inhibitors ◽  
Resistant Mutants ◽  
New Compounds ◽  
Susceptibility Profiles ◽  
Dna Substrate ◽  
Hiv 1

ABSTRACT Integrase strand transfer inhibitors (INSTIs) have emerged as clinically effective therapeutics that inhibit HIV-1 replication by blocking the strand transfer reaction catalyzed by HIV-1 integrase (IN). Of the three FDA-approved INSTIs, dolutegravir (DTG) is the least apt to select for resistance. However, recent salvage therapy regimens had low response rates with therapies that included DTG, suggesting that DTG resistance can be selected in patients. Using a single-round infection assay, we evaluated a collection of our best inhibitors and DTG against a broad panel of INSTI-resistant mutants. Two of the new compounds, 4c and 4d, had antiviral profiles against the mutants we tested superior to that of DTG. The susceptibility profiles of 4c and 4d suggest that the compounds are candidates for development as INSTIs. Modeling the binding of 4d to HIV-1 IN reinforced the significance of mimicking the DNA substrate in developing compounds that are broadly effective in their abilities to inhibit HIV-1 INs with mutations in the active site.

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