Elvitegravir

Manufactured by Gilead Sciences

Sourced in India, United States, Spain

Elvitegravir is an integrase strand transfer inhibitor (INSTI) used in the treatment of HIV-1 infection. It is a laboratory compound developed by Gilead Sciences for research purposes.

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Lab products found in correlation

9 protocols using elvitegravir

Characterization of HIV-1 Viral Proteins

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Rabbit polyclonal antisera against HIV-1 CA, MA, NC, or IN were raised against purified recombinant proteins. Mouse monoclonal antibody 414, recognizing phenylalanine–glycine repeats of nuclear pore complex proteins, was from Abcam (UK); mouse monoclonal antibody against cytochrome C was from BD Biosciences (Franklin, NJ); affinity purified rabbit antibody against CPSF6 was obtained from Sigma (HPA039973). Secondary antibodies Alexa Fluor 405 Goat Anti-Mouse IgG, Alexa Fluor 532 Goat Anti-Rabbit IgG, and Alexa Fluor 568 Goat Anti-Rabbit IgG were from Life Technologies.
Stock solutions of 6 mM aphidicolin (Sigma A0781), 5 mM efavirenz (obtained through the AIDS Research and Reference Reagent Program, Division of AIDS, NIAID), 10 mM PF74 (Gilead Sciences, Foster City, CA), and 10 mM elvitegravir (Gilead Sciences), respectively, were prepared in dimethyl sulfoxide (DMSO) and stored at −20°C.

Peng K., Muranyi W., Glass B., Laketa V., Yant S.R., Tsai L., Cihlar T., Müller B, & Kräusslich H.G. (2014). Quantitative microscopy of functional HIV post-entry complexes reveals association of replication with the viral capsid. eLife, 3, e04114.

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Elvitegravir Formulation and Characterization

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Elvitegravir was provided by Gilead Sciences (Foster City, CA) via CONRAD (Arlington, VA). PLA-HPG was synthesized as previously described [25 (link)]. IR-780 iodide, glycerol, NaIO₄, Na₂SO₃, bovine serum albumin (BSA), lactic acid, acetic acid, mucin, urea, glucose, Tween 80, Solutol (Kolliphor) HS 15, and HPLC-grade formic acid were obtained from Sigma-Aldrich. The 4-(4-(dihexadecylamino)styryl)-N-methylpyridinium iodide salt (DiA) and 4,6-diamidino-2 phenylindole (DAPI) stain were ordered from Invitrogen. Antibody stains anti-CD45 (ab10558) and anti-EpCAM (ab71916) and corresponding isotype controls for flow cytometry were purchased from Abcam. HPLC grade acetonitrile and water were purchased from VWR (J.T. Baker).

Mohideen M., Quijano E., Song E., Deng Y., Panse G., Zhang W., Clark M.R, & Saltzman W.M. (2017). Degradable Bioadhesive Nanoparticles for Prolonged Intravaginal Delivery and Retention of Elvitegravir. Biomaterials, 144, 144-154.

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Antiretroviral Drug Effects on Cell Viability

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The following chemicals/drugs were used: zidovudine (GlaxoSmithKline), stavudine (Bristol-Myers Squibb), tenofovir disoproxil fumarate (Gilead Sciences), nevirapine (Boehringer Ingelheim), efavirenz (Bristol-Myers Squibb), rilpivirine (Janssen Pharmaceuticals, Inc.), ritonavir (Abbott Laboratories), lopinavir (Abbott Laboratories), nelfinavir (Agouron), atazanavir (Bristol-Myers Squibb), raltegravir (sc-364600; Santa Cruz), elvitegravir (Gilead Sciences), maraviroc (Pfizer), tunicamycin (T7765; Sigma-Aldrich Biotechnology), thapsigargin (T9033; Sigma-Aldrich), 4-phenylbutyric acid ([PBA] P21005; Sigma-Aldrich), 6-hydroxy-2,5,7,8-tetramethylchroman-2-carboxylic acid ([Trolox] 648471; Calbiochem), and rosiglitazone (ALX-350-125; Alexis Biochemicals). The reagents for cell culture were from Sigma-Aldrich, whereas media and FBS were from Life Technologies. For initial assessments of the effects of antiretroviral drugs prior to concentration-response analyses, the drugs were used at the highest concentrations that did not cause cytotoxicity in specific cell systems. The drugs were dissolved using dimethyl sulfoxide (DMSO) as the vehicle. Controls included amounts of DMSO (≤0.1% DMSO of total cell medium volume) equal to those used in drug-treated cells.

Moure R., Domingo P., Villarroya J., Gasa L., Gallego-Escuredo J.M., Quesada-López T., Morón-Ros S., Maroto A.F., Mateo G.M., Domingo J.C., Villarroya F, & Giralt M. (2018). Reciprocal Effects of Antiretroviral Drugs Used To Treat HIV Infection on the Fibroblast Growth Factor 21/β-Klotho System. Antimicrobial Agents and Chemotherapy, 62(6), e00029-18.

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HIV Drug Combination Evaluation

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Tenofovir disoproxil fumarate (TDF), emtricitabine (FTC), and elvitegravir (EVG) kindly were provided by Gilead Sciences, Inc (Foster City, CA), or purchased from Selleck Chemicals LLC (Houston, TX). Maraviroc (MVC) was purchased from Selleck Chemicals LLC. The peptide C5A (purity ≥ 80%) was obtained from GenScript USA, Inc (Piscataway, NJ). All other reagents were obtained from Sigma-Aldrich (St. Louis, MO), unless otherwise noted.

Gallay P.A., Ramirez C.M, & Baum M.M. (2023). Acute antagonism in three-drug combinations for vaginal HIV prevention in humanized mice. Scientific Reports, 13, 4594.

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Formulation and Characterization of Anti-HIV Suppositories

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Tenofovir (TFV) used in the formulations was purchased from Wuxi, AppTec (Shanghai, China). Tenofovir alafenamide fumarate (TAF) was provided by Laurus Labs to the Centers for Disease Control and Prevention (CDC) (Atlanta, GA, USA). Elvitegravir (EVG) was provided by Gilead Sciences, Foster City, CA (for the TFV + EVG suppositories) or by Hetero Drugs (Hyderabad, India) to CDC (for the TAF + EVG suppositories). The fat-soluble base witepsol was a gift from IOI Oleo GmbH (Hamburg, Germany), and suppocire base was from Gattefosse (Paramus, NJ, USA). Cocoa butter and three Polyethylene-Glycol (PEG) polymers of different molecular weights of 3350, 1000, 400 were purchased from Spectrum Chemical, Gardena, CA, USA. The plastic disposable suppository molds were purchased from Professional Compounding Centers of America (PCCA), Houston, TX, USA. Tetra-butyl ammonium bromide, potassium mono- and di-phosphate salts used in the HPLC assays were purchased from Fisher Scientific, NJ, or J.T. Baker, NJ.

Jhunjhunwala K., Dobard C.W., Sharma S., Makarova N., Holder A., Dinh C., Mitchell J., Wang L., Zhang J., Patel S.K., Heneine W, & Rohan L.C. (2021). Development, Characterization and In Vivo Pharmacokinetic Assessment of Rectal Suppositories Containing Combination Antiretroviral Drugs for HIV Prevention. Pharmaceutics, 13(8), 1110.

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Antiretroviral Drug Sources for Research

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The antiretroviral drugs used in this study were obtained from the following sources: zidovudine, AZT; didanosine, ddI; stavudine, d4T; lamivudine, 3TC; abacavir, ABC; tenofovir, TDF; emtricitabine, FTC; nevirapine, NVP; delavirdine, DLV; efavirenz, EFV; etravirine, ETR; saquinavir, SQV; ritonavir, RTV; indinavir, IDV; nelfinavir, NFV; amprenavir, APV; lopinavir, LPV; atazanavir, ATV; tipranavir, TPV; and darunavir, DRV (ENZO Life Sciences International, Inc., Plymouth Meeting, PA, formerly BioMol International, LP); raltegravir, RAL and elvitegravir, EVG (Gilead Sciences, Inc., Foster City, CA).

Gibson R.M., Weber J., Winner D., Miller M.D, & Quiñones-Mateu M.E. (2014). Contribution of Human Immunodeficiency Virus Type 1 Minority Variants to Reduced Drug Susceptibility in Patients on an Integrase Strand Transfer Inhibitor-Based Therapy. PLoS ONE, 9(8), e104512.

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Comprehensive Synthesis of Antiviral Compounds

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GS-9770, atazanavir (ATV), darunavir (DRV), amprenavir (APV), bictegravir (BIC), elvitegravir (EVG), lenacapavir (LEN), tenofovir alafenamide (TAF), emtricitabine (FTC), efavirenz (EFV), stavudine (d4T), the RSV control inhibitor YM-53403, and the SARS CoV-2 control inhibitor remdesivir (RDV) were all synthesized at Gilead Sciences (Foster City, CA). Ribavirin (RBV) was purchased from Combi-Blocks (San Diego, CA). The protein synthesis inhibitor puromycin and the HIV-positive control azidothymidine (AZT) were purchased from Sigma-Aldrich (St. Louis, MO). The HBV control inhibitor ND-654 was purchased from Pharmaron (Beijing, China). The HCV protease inhibitor danoprevir was purchased from Selleck Chemicals (Houston, TX). All drug stocks were prepared in 100% dimethyl sulfoxide (DMSO) and stored frozen at −20°C.

Mulato A., Lansdon E., Aoyama R., Voigt J., Lee M., Liclican A., Lee G., Singer E., Stafford B., Gong R., Murray B., Chan J., Lee J., Xu Y., Ahmadyar S., Gonzalez A., Cho A., Stepan G.J., Schmitz U., Schultz B., Marchand B., Brumshtein B., Wang R., Yu H., Cihlar T., Xu L, & Yant S.R. (2024). Preclinical characterization of a non-peptidomimetic HIV protease inhibitor with improved metabolic stability. Antimicrobial Agents and Chemotherapy, 68(4), e01373-23.

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HIV Drug Resistance Profiling Protocol

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Dolutegravir was provided by ViiV Healthcare Ltd (Research Triangle Park, NC, USA).
Elvitegravir was obtained from Gilead Sciences Inc. (Foster City, CA, USA). Lamivudine was
obtained from the NIH AIDS Research and Reference Reagent Program (http://www.aidsreagent.org). HIV-1 strains from 6 and 11 patients from
singleton/small and large clusters, respectively, were used to infect CBMCs at a similar
moi of 0.1, i.e. 1 × 10⁶ TCID₅₀ per 10 × 10⁶ CBMCs, based
on p24 and RT assays.⁵⁷ (link) Viruses
were serial passaged in CBMCs in the presence of escalating concentrations of
dolutegravir, Elvitegravir and/or lamivudine over 24–36 weeks, as previously
described.⁵⁸ (link)^,⁵⁹ (link) RT activity in culture
supernatants was quantified weekly to monitor viral replication of individual viral
variants in the presence and absence of drug pressure, adjusting incremental increases in
drug concentrations if RT activity assays exceeded 100 000 cpm.⁵⁸ (link)^,⁵⁹ (link) At each passage, aliquots of cell-free supernatant were stored
at −70°C for further genotypic analysis.

Brenner B.G., Ibanescu R.I., Oliveira M., Roger M., Hardy I., Routy J.P., Kyeyune F., Quiñones-Mateu M.E, & Wainberg M.A. (2017). HIV-1 strains belonging to large phylogenetic clusters show accelerated escape from integrase inhibitors in cell culture compared with viral isolates from singleton/small clusters. Journal of Antimicrobial Chemotherapy, 72(8), 2171-2183.

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Comprehensive Antiretroviral Drug Evaluation

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A set of antiretroviral drugs representative of each antiretroviral class were included (Table 1) in the study. All antiretroviral drugs were purchased from Merck Life Science (Madrid, Spain), except for Dolutegravir (DTG), Doravirine (DOR), Rilpivirine (RIL) and Cobicistat (COB) which were purchased from Quimigen (Madrid, Spain) and Tenofovir alafenamide (TAF), Bictegravir (BIC) and Elvitegravir (EVG) which were kindly provided by Gilead Sciences Inc. (Foster City, California, USA). Stock solutions of each ARV were prepared using dimethyl sulfoxide (DMSO), deionized water or methanol according to each ARV solubility (Table 1). Additionally, active antibiotics against the analyzed bacterial strains including Colistin, Gentamicin and Clindamycin were purchased from Quimigen (Madrid, Spain).

Rubio-Garcia E., Ferrando N., Martin N., Ballesté-Delpierre C., Miró J.M., Paredes R., Casals-Pascual C, & Vila J. (2024). In vitro antibacterial activity of antiretroviral drugs on key commensal bacteria from the human microbiota. Frontiers in Cellular and Infection Microbiology, 13, 1306430.

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