Balzarini J, Karlsson A, Pérez-Pérez M J, Camarasa M J, Tarpley W G, De Clercq E
Rega Institute for Medical Research, Katholieke Universiteit Leuven, Belgium.
J Virol. 1993 Sep;67(9):5353-9. doi: 10.1128/JVI.67.9.5353-5359.1993.
Human immunodeficiency virus type 1 (HIV-1)-infected CEM cells were treated by the HIV-1-specific inhibitors bis-heteroarylpiperazine (BHAP), 4,5,6,7-tetrahydro-5-methylimidazo[4,5,1-jk][1,4]benzodiazepin-2(1 H)-on e (TIBO) R82913, nevirapine, and the N3-methylthymine derivative of [2',5'-bis-O-(tert-butyldimethylsilyl)-beta-D-ribofuranosyl]-3'-spiro- 5''-(4''-amino-1'',2''-oxathiole-2'',2''-dioxide) (TSAO-m3T), as single agents or in combination, at escalating concentrations. When used individually, the compounds led to the emergence of drug-resistant virus strains within two to five subcultivations. The resulting strains were designated HIV-1/BHAP, HIV-1/TIBO, HIV-1/Nev, and HIV-1/TSAO-m3T, respectively. The mutant viruses showed the following amino acid substitutions in their reverse transcriptase (RT): Leu-100-->Ile for HIV-1/BHAP; Lys-103-->Asn for HIV-1/TIBO; Val-106-->Ala for HIV-1/Nev; and Glu-138-->Lys for HIV-1/TSAO-m3T. Both the Tyr-181-->Cys and Val-106-->Ala mutations were found in another mutant emerging following treatment with nevirapine at escalating concentrations. The BHAP-resistant virus remained fully sensitive to the inhibitory effects of nevirapine and TSAO-m3T, whereas the TSAO-m3T-resistant virus remained fully sensitive to the inhibitory effects of nevirapine and BHAP. When different pairs of nonnucleoside RT inhibitors (i.e., BHAP plus TSAO-m3T, nevirapine plus TSAO-m3T, TIBO plus TSAO-m3T, nevirapine plus TIBO, and BHAP plus nevirapine) were used, resistant virus emerged as fast as with single-drug therapy. In all cases the Tyr-181-->Cys mutation appeared; the virus showed markedly reduced sensitivity to all HIV-1-specific inhibitors but retained sensitivity to 2',3'-dideoxynucleoside analogs such as zidovudine, ddC, and ddI. Our findings argue against simultaneous combination of two different nonnucleoside RT inhibitors that are unable to inhibit HIV-1 mutant strains containing the Tyr-181-->Cys mutation when administered as single drugs.
用1型人类免疫缺陷病毒（HIV-1）特异性抑制剂双杂芳基哌嗪（BHAP）、4,5,6,7-四氢-5-甲基咪唑并[4,5,1-jk][1,4]苯并二氮杂䓬-2(1H)-酮（TIBO）R82913、奈韦拉平以及[2',5'-双-O-(叔丁基二甲基甲硅烷基)-β-D-呋喃核糖基]-3'-螺-5''-(4''-氨基-1'',2''-氧硫杂环戊烯-2'',2''-二氧化物)（TSAO-m3T）的N3-甲基胸腺嘧啶衍生物，以递增浓度单独或联合处理HIV-1感染的CEM细胞。单独使用时，这些化合物在两到五次传代培养内导致耐药病毒株出现。所得毒株分别命名为HIV-1/BHAP、HIV-1/TIBO、HIV-1/Nev和HIV-1/TSAO-m3T。突变病毒在其逆转录酶（RT）中出现以下氨基酸取代：HIV-1/BHAP的Leu-100→Ile；HIV-1/TIBO的Lys-103→Asn；HIV-1/Nev的Val-106→Ala；HIV-1/TSAO-m3T的Glu-138→Lys。在用递增浓度奈韦拉平处理后出现的另一种突变体中发现了Tyr-181→Cys和Val-106→Ala两种突变。对BHAP耐药的病毒对奈韦拉平和TSAO-m3T的抑制作用仍完全敏感，而对TSAO-m3T耐药的病毒对奈韦拉平和BHAP的抑制作用仍完全敏感。当使用不同组合的非核苷类RT抑制剂（即BHAP加TSAO-m3T、奈韦拉平加TSAO-m3T、TIBO加TSAO-mT、奈韦拉平加TIBO以及BHAP加奈韦拉平）时，耐药病毒出现的速度与单药治疗时一样快。在所有情况下都出现了Tyr-181→Cys突变；该病毒对所有HIV-1特异性抑制剂的敏感性显著降低，但对齐多夫定、双脱氧胞苷和双脱氧肌苷等核苷类似物仍保持敏感性。我们的研究结果表明，两种不同的非核苷类RT抑制剂不能抑制含有Tyr-181→Cys突变的HIV-1突变株，因此不宜同时联合使用。
