Screening for Antiviral Inhibitors of the HIV Integrase—LEDGF/p75 Interaction Using the AlphaScreen™ Luminescent Proximity Assay

Small-molecule inhibitors of HIV integrase (HIV IN) have emerged as a promising new class of antivirals for the treatment of HIV/AIDS. The compounds currently approved or in clinical development specifically target HIV DNA integration and were identified using strand-transfer assays targeting the HIV IN/viral DNA complex. The authors have developed a second biochemical assay for identification of HIV integrase inhibitors, targeting the interaction between HIV IN and the cellular cofactor LEDGF/p75. They developed a luminescent proximity assay (AlphaScreen™) designed to measure the association of the 80-amino-acid integrase binding domain of LEDGF/p75 with the 163-amino-acid catalytic core domain of HIV IN. This assay proved to be quite robust (with a Z′ factor of 0.84 in screening libraries arrayed as orthogonal mixtures) and successfully identified several compounds specific for this protein-protein interaction. ( Journal of Biomolecular Screening 2008:406-414)

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Quinoline-based Protein–protein Interaction Inhibitors of LEDGF/p75 and HIV Integrase: An In Silico Study

Current Topics in Medicinal Chemistry ◽

10.2174/1568026619666190208164801 ◽

2019 ◽

Vol 18 (32) ◽

pp. 2800-2815 ◽

Cited By ~ 1

Author(s):

Nisha Chhokar ◽

Sourav Kalra ◽

Monika Chauhan ◽

Anjana Munshi ◽

Raj Kumar

Keyword(s):

Protein Interactions ◽

Biological Data ◽

Strand Transfer ◽

Hiv Integrase ◽

Protein Protein Interactions ◽

Allosteric Site ◽

Protein Protein Interaction ◽

In Silico Study ◽

Cellular Cofactor

The failure of the Integrase Strand Transfer Inhibitors (INSTIs) due to the mutations occurring at the catalytic site of HIV integrase (IN) has led to the design of allosteric integrase inhibitors (ALLINIs). Lens epithelium derived growth factor (LEDGF/p75) is the host cellular cofactor which helps chaining IN to the chromatin. The protein-protein interactions (PPIs) were observed at the allosteric site (LEDGF/p75 binding domain) between LEDGF/p75 of the host cell and IN of virus. In recent years, many small molecules such as CX04328, CHIBA-3053 and CHI-104 have been reported as LEDGF/p75-IN interaction inhibitors (LEDGINs). LEDGINs have emerged as promising therapeutics to halt the PPIs by binding at the interface of both the proteins. In the present work, we correlated the docking scores for the reported LEDGINs containing quinoline scaffold with the in vitro biological data. The hierarchal clustering method was used to divide the compounds into test and training set. The robustness of the generated model was validated by q2 and r2 for the predicted set of compounds. The generated model between the docking score and biological data was assessed to predict the activity of the hits (quinoline scaffold) obtained from virtual screening of LEDGINs providing their structureactivity relationships to aim for the generation of potent agents.

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Update on HIV integrase inhibitors for the treatment of HIV-1 infection

Future Virology ◽

10.2217/fvl-2019-0077 ◽

2019 ◽

Vol 14 (10) ◽

pp. 693-709

Author(s):

Blake Max

Keyword(s):

Treatment Guidelines ◽

Hiv Treatment ◽

Strand Transfer ◽

Hiv Integrase ◽

New Class ◽

Treatment Paradigm ◽

Short And Long Term ◽

Hiv Integrase Inhibitors ◽

Hiv 1

It has been over 30 years since the first antiretroviral agent was approved for treatment of HIV-1 infection and its impact on morbidity and mortality has been dramatic. However, early treatments were hindered by short- and long-term toxicity, poor tolerability, high pill burden, drug interactions and development of drug resistance. A major breakthrough in HIV therapeutics occurred over a decade ago with a new class of drugs that not only are preferred by HIV treatment guidelines but also are changing the HIV treatment paradigm. This new class of drugs are called HIV-1 integrase strand transfer inhibitors and they have established a role in almost every aspect of HIV treatment.

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Targeting Integration beyond Strand Transfer: Development of Second-Generation HIV Integrase Inhibitors

Antiviral Drug Strategies - Methods and Principles in Medicinal Chemistry ◽

10.1002/9783527635955.ch3 ◽

2011 ◽

pp. 51-71

Author(s):

Arnout R. D. Voet ◽

Marc De Maeyer ◽

Frauke Christ ◽

Zeger Debyser

Keyword(s):

Second Generation ◽

Strand Transfer ◽

Hiv Integrase ◽

Integrase Inhibitors ◽

Hiv Integrase Inhibitors

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Unexpected Isomerisation of a Fragment Analogue During Fragment-Based Screening of HIV Integrase Catalytic Core Domain

Australian Journal of Chemistry ◽

10.1071/ch15587 ◽

2015 ◽

Vol 68 (12) ◽

pp. 1871

Author(s):

John H. Ryan ◽

Karen E. Jarvis ◽

Roger J. Mulder ◽

Craig L. Francis ◽

G. Paul Savage ◽

...

Keyword(s):

Human Immunodeficiency Virus ◽

Binding Site ◽

Hiv Integrase ◽

Catalytic Core Domain ◽

Aromatic Carboxylic Acid ◽

Catalytic Core ◽

Core Domain ◽

Acid Fragment ◽

Immunodeficiency Virus

Fragment-based screening of human immunodeficiency virus type 1 (HIV) integrase revealed several aromatic carboxylic acid fragment hits, some of which bound weakly at the site on the HIV-integrase catalytic core domain that binds the lens epithelium-derived growth factor (LEDGF). Virtual screening of an internal database identified an analogue that bound with higher affinity and in an isomerised form to the LEDGF binding site. The starting lactone was stable in CDCl3; however, an unexpected isomerisation process occurred in [D6]DMSO to give the same isomer found in the LEDGF binding site. This hit led directly to a series of low-micromolar LEDGF inhibitors and, via a scaffold hop, to a series of allosteric binding site inhibitors.

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Peptides Derived from the Reverse Transcriptase of Human Immunodeficiency Virus Type 1 as Novel Inhibitors of the Viral Integrase

Journal of Biological Chemistry ◽

10.1074/jbc.m414679200 ◽

2005 ◽

Vol 280 (23) ◽

pp. 21987-21996 ◽

Cited By ~ 45

Author(s):

Iris Oz Gleenberg ◽

Orna Avidan ◽

Yehuda Goldgur ◽

Alon Herschhorn ◽

Amnon Hizi

Keyword(s):

Reverse Transcriptase ◽

Catalytic Domain ◽

Three Dimensional ◽

Specific Protein ◽

Strand Transfer ◽

Catalytic Core ◽

Core Domain ◽

Inhibitory Peptides ◽

Hiv 1

Recent studies have shown that the integrase (IN) of HIV-1 is inhibited in vitro by HIV-1 reverse transcriptase (RT). We further investigated the specific protein sequences of RT that were involved in this inhibition by screening a complete library of RT-derived peptides for their inhibition of IN activities. Two 20-residue peptides, peptide 4286, derived from the RT DNA polymerase domain, and the one designated 4321, from the RT ribonuclease H domain, inhibit the enzymatic activities of IN in vitro. The former peptide inhibits all three IN-associated activities (3′-end processing, strand transfer, and disintegration), whereas the latter one inhibits primarily the first two functions. We showed the importance of the sequences and peptide length for the effective inhibition of IN activities. Binding assays of the peptides to IN (with no DNA substrate present) indicated that the two inhibitory peptides (as well as several non-inhibitory peptides) interact directly with IN. Moreover, the isolated catalytic core domain of IN also interacted directly with the two inhibitory peptides. Nevertheless, only peptide 4286 can inhibit the disintegration activity associated with the IN core domain, because this activity is the only one exhibited by this domain. This result was expected from the lack of inhibition of disintegration of full-length IN by peptide 4321. The data and the three-dimensional models presented suggested that the inhibition resulted from steric hindrance of the catalytic domain of IN. This information can substantially facilitate the development of novel drugs against HIV INs and thus contribute to the fight against AIDS.

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HIV integrase inhibitors that inhibit strand transfer interact with RAG1 and hamper its activities

International Immunopharmacology ◽

10.1016/j.intimp.2021.107515 ◽

2021 ◽

Vol 95 ◽

pp. 107515

Author(s):

Namrata M. Nilavar ◽

Sathees C. Raghavan

Keyword(s):

Strand Transfer ◽

Hiv Integrase ◽

Integrase Inhibitors ◽

Hiv Integrase Inhibitors

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Long Dissociation of Bictegravir from HIV-1 Integrase-DNA Complexes

Antimicrobial Agents and Chemotherapy ◽

10.1128/aac.02406-20 ◽

2021 ◽

Vol 65 (5) ◽

Author(s):

Kirsten L. White ◽

Nathan Osman ◽

Ernesto Cuadra-Foy ◽

Bluma G. Brenner ◽

Devleena Shivakumar ◽

...

Keyword(s):

Antiviral Activity ◽

Ring System ◽

Strand Transfer ◽

Hiv Integrase ◽

Wild Type ◽

Resistance Mutations ◽

Dna Complexes ◽

Transfer Inhibitor ◽

Dna Complex ◽

Hiv 1

ABSTRACT The HIV integrase (IN) strand transfer inhibitor (INSTI) bictegravir (BIC) has a long dissociation half-life (t1/2) from wild-type IN-DNA complexes: BIC 163 h > dolutegravir (DTG) 96 h > raltegravir (RAL) 10 h > elvitegravir (EVG) 3.3 h. In cells, BIC had more durable antiviral activity against wild-type HIV after drug washout than RAL or EVG. BIC also had a longer t1/2 and maintained longer antiviral activity after drug washout than DTG with the clinically relevant resistance IN mutant G140S+Q148H. Structural analyses indicate that BIC makes more contacts with the IN-DNA complex than DTG mainly via its bicyclic ring system, which may contribute to more prolonged residence time and resilience against many resistance mutations.

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