Characterization of a Mutant HIV-1 Reverse Transcriptase Resistant to (+)-(5S)-4,5,6,7-tetrahydro-5-methyl-6-(3-methyl-2-butenyl)-imidazo[4,5,1- jk ][1,4]benzodiazepin-2(1 H )-thione (TIBO R82150)
Free accessBrief reportFirst published online August, 1994
Characterization of a Mutant HIV-1 Reverse Transcriptase Resistant to (+)-(5S)-4,5,6,7-tetrahydro-5-methyl-6-(3-methyl-2-butenyl)-imidazo[4,5,1- jk ][1,4]benzodiazepin-2(1 H )-thione (TIBO R82150)
A virus strain resistant to R82150, a non-nucleoside reverse transcriptase (NNRT) inhibitor (tetrahydro-imidazo [4,5, 1-jk] [1,4] benzodiazepine-2(1H)-thione), was isolated following serial passage of HIV-1 RF in CEM-SS cells. The virus is cross-resistant to another non-nucleoside reverse transcriptase inhibitor, TGG-II-23A [1,4-dimethyl-1-[5,5-dimethyl-2-oxazoionyl]-naphthalen-2-one), but remains susceptible to AZT, DDI, D4T and phosphonoformate (PFA). DNA sequencing of reverse transcriptase genes from resistant virus indicated that R82150 selects for amino acid alterations Y181C and V108I. In vitro mutagenized reverse transcriptase and recombinant HIV-1 (pNL4-3) carrying either of the mutations have been generated. Genotypic and phenotypic analyses identified V108I as an unreported R82150-associated mutation. Both reverse transcriptase and viral resistance assays indicated that the resistance conferred by the V108I mutation is 7-fold less than that conferred by Y181C.
References
1.
AlamM.BechtoldC.M.PatickA.K.SkoogM.T.GantT.G.ColonnoR.J.MyersA.I.LiH.TrimbleJ., and LinP.-F. (1993) Substituted naphthalenones as a new structural class of HIV-1 reverse transcriptase inhibitors. Antiviral Res22: 131–141.
2.
BabaM.De ClercqE.TanakaH.UbasawaM.TakashimaH.SekiyaK.NittaI.UmezuK.NakashimaH.MoriS.ShigetaS.WalkerR.T., and MiyasakaT. (1991) Potent and selective inhibition of human immunodeficiency virus type 1 (HIV-1) by 5-ethyl-6-phenylthiouracil derivatives through their interaction with the HIV-1 reverse transcriptase. Proc Natl Acad Sci USA88: 2356–2360.
3.
BalzariniJ.KarlssonA.Perez-PerezM.-J.VrangL.Wal-BergsJ.ZhangH.ÖbergB.VandammeA.-M.CamarasaM.-J., and De ClercqE. (1993) HIV-1-specific reverse transcriptase inhibitors show differential activity against HIV-1 mutant strains containing different amino acid substitutions in the reverse transcriptase. Virology192: 246–253.
4.
BalzariniJ.Pèrez-PèrezM.-J.San-FèlixA.ScholsD.PemoC.-F.VandammeA.-M.CamarasaM.-J., and De ClercqE. (1992) 2′,5′-Bis-0-flert-butyldimethylsilyl)-3′spiro 5′-(4″-amino-1″,2″-oxathiole-2″,2″-dioxide)pyrimidine (TSAO) nucleoside analogues: Highly selective inhibitors of human immunodeficiency virus type 1 that are targeted at the viral reverse transcriptase. Proc Natl Acad Sci USA89: 4392–4396.
5.
ByrnesV.W.SardanaV.V.SchleifW.A.CondraJ.H.WaterburyJ.A.WolfgangJ.A.LongW.J.SchneiderC.L.SchlabachA.J.WolanskiB.S.GrahamD.J.GotlibLRhodesA.TitusD.L.RothE.BlahyO.M.QuinteroJ.C.StaszewskiS., and EminiE.A. (1993) Comprehensive mutant enzyme and viral variant assessment of human immunodeficiency virus type 1 reverse transcriptase resistance to nonnucleoside inhibitors. Antimicrob Agents Chemother37: 1576–1579.
6.
De ClercqE. (1992) HIV inhibitors targeted at the reverse transcriptase. AIDS Res & Human Retro8: 119–134.
7.
DengW.P., and NickoloffJ.A. (1992) Site directed mutagenesis of virtually any plasmid by eliminating a unique site. Anal Biochem200: 81–88.
8.
GoldmanM.E.NunbergJ.H.O'BrienJ.A.QuinteroJ.C.SchleifW.A.FreundK.F.GaulS.L.SaariW.S.WaiJ.S.HoffmanJ.M.AndersonP.S.HupeD.J.EminiE.A., and SternA.M. (1991) Pyridinone derivatives: Specific human immunodeficiency virus type 1 reverse transcriptase inhibitors with antiviral activity. Proc Natl Sci USA88: 6863–6867.
9.
GrobP.M.WuJ.C.CohenK.A.IngrahamR.H.ShihC-K.HargraveK.D.McTagueT.L., and MerluzziV.J. (1992) Non-nucleoside inhibitors of HIV-1 reverse transcriptase: Nevirapine as prototype drug. AIDS Res Human Retro8: 145–152.
10.
JohnsonV.A.ByingtonR.E., and KaplanJ.C. (1990) Reverse transcriptase (RT) activity assay. In Techniques in HIV Research.AldoviniA., and WalkerB. (eds). New York: Stockton Press, pp. 98–102.
11.
LarderB.A., and KempS.D. (1989) Multiple mutations in HIV-1 reverse transcriptase confer high level resistance to zidovdine (AZT). Science246: 1155–1158.
12.
LarderB.A. (1992) 3′-azido-3′deoxythymidine resistance suppressed by a mutation conferring human immunodeficiency virus type 1 resistance to nonnucleoside reverse transcriptase inhibitors. Antimicrob Ag Chemother36: 2664–2669.
13.
MellorsJ.W.DutschmanG.E.ImG.-J.TramontanoE.WinklerS.R., and ChengY.-C. (1992) In vitro selection and molecular characterization of human immunodeficiency virus-1 resistant to non-nucleoside inhibitors of reverse transcriptase. Mol Pharm41: 446–451.
14.
MellorsJ.W.ImG.-J.TramontanoE.WinklerS.R.MedinaD.J.DutschmanG.E.BazmiH.Z.PirasG.GonzalezC.J., and ChengY.-C. (1993) A single conservative amino acid substitution in the reverse transcriptase of human immunodeficiency virus-1 confers resistance to (+)-(5S)-4,5,6,7-tetrahydro-5methyl-6(3-methyl-2-butenyl)-imidazo (4,5,1-jk)(1,4)benzodiazepin-2(1H)-thione (TIBO R82150). Mol Pharm43: 11–16.
15.
MerluzziV.J.HargraveK.D.LabadiaM.GrozingerK.SkoogM.WuJ.C.ShihC.K.EcknerK.HattoxS.AdamsJ.RosehthalA.S.FaanesR.EcknerR.J.KoupR.A., and SullivanJ.L. (1990) Inhibition of HIV-1 replication by a nonnucleoside reverse transcriptase inhibitor. Science250: 1411–1413.
16.
NunbergJ.H.SchleifW.A.BootsE.J.O'BrienJ.A.QuinteroJ.C.HoffmanJ.M.EminiE.A., and GoldmanM.E. (1991) Viral resistance to human immunodeficiency virus type 1-specific pyridinone reverse transcriptase inhibitors. J Virol65: 4887–4892.
17.
PauwelsR.AndriesK.DesmyterJ.ScholsD.KuklaM.J.BreslinH.J.RaeymaeckersA.GelderJ.V.WoestenborghsR.HeykantsJ.SchellekensK.JanssenM.A.C.De ClercqE., and JanssenP.A.J. (1990) Potent and selective inhibition of HIV-1 replication in vitro by a novel series of TIBO derivatives. Nature343: 470–474.
18.
RichmanD.D. (1993). Playing chess with reverse transcriptase. Nature361: 588–589.
19.
RichmanD.ShihC.-K.LowyI.RoseJ.ProdanovichP.GoffS., and GriffinJ. (1991) Human immunodeficiency virus type 1 mutants resistant to nonnucleoside inhibitors of reverse transcriptase arise in tissue culture. Proc Natl Acad Sci USA88: 11241–11245.
20.
RomeroD.L.BussoM.TanC.-K.ReusserF.PalmerJ.R.PoppeS.M.AristoffP.A.DowneyK.M.SoA.G.ResnickL., and TarpleyW.G. (1991) Nonnucleoside reverse transcriptase inhibitors that potently and specifically block human immunodeficiency virus type 1 replication. Proc Natl Acad Sci USA88: 8806–8810.
21.
SoudeynsH.YaoX.-J.GaoQ.BelleauB.KrausJ.-L.Nguyen-BaN.SpiraB., and WainbergM.A. (1991). Anti-human immunodeficiency virus type I activity and in vitro toxicity 2′-deoxy-3′-thiacytidine (BCH-189), a novel heterocyclic nucleoside analog. Antimicro Agents & Chemo35: 1386–1390.
22.
TramontanoE., and ChengY.-C. (1992) HIV-1 Reverse transcriptase inhibition by a dipyridodiazepinone derivative: BI***-RG-587. Biochemical Pharmacology43: 1371–1376.
23.
WeislowO.S.KiserR.FineD.L.BaderJ.ShoemakerR.H., and BoydM.R. (1989) New soluble-formazan assay for HIV-1 cytopathic effects: application to high-flux screening of synthetic and natural products for AIDS-antiviral activity. J Natl Can Inst81: 577–586.
