Vingerhoets Johan, Azijn Hilde, Fransen Els, De Baere Inky, Smeulders Liesbet, Jochmans Dirk, Andries Koen, Pauwels Rudi, de Béthune Marie-Pierre
Tibotec, Mechelen, Belgium.
J Virol. 2005 Oct;79(20):12773-82. doi: 10.1128/JVI.79.20.12773-12782.2005.
TMC125 is a potent new investigational nonnucleoside reverse transcriptase inhibitor (NNRTI) that is active against human immunodeficiency virus type 1 (HIV-1) with resistance to currently licensed NNRTIs. Sequential passage experiments with both wild-type virus and NNRTI-resistant virus were performed to identify mutations selected by TMC125 in vitro. In addition to "classic" selection experiments at a low multiplicity of infection (MOI) with increasing concentrations of inhibitors, experiments at a high MOI with fixed concentrations of inhibitors were performed to ensure a standardized comparison between TMC125 and current NNRTIs. Both low- and high-MOI experiments demonstrated that the development of resistance to TMC125 required multiple mutations which frequently conferred cross-resistance to efavirenz and nevirapine. In high-MOI experiments, 1 muM TMC125 completely inhibited the breakthrough of resistant virus from wild-type and NNRTI-resistant HIV-1, in contrast to efavirenz and nevirapine. Furthermore, breakthrough of virus from site-directed mutant (SDM) SDM-K103N/Y181C occurred at the same time or later with TMC125 as breakthrough from wild-type HIV-1 with efavirenz or nevirapine. The selection experiments identified mutations selected by TMC125 that included known NNRTI-associated mutations L100I, Y181C, G190E, M230L, and Y318F and the novel mutations V179I and V179F. Testing the antiviral activity of TMC125 against a panel of SDMs indicated that the impact of these individual mutations on resistance was highly dependent upon the presence and identity of coexisting mutations. These results demonstrate that TMC125 has a unique profile of activity against NNRTI-resistant virus and possesses a high genetic barrier to the development of resistance in vitro.
TMC125是一种新型强效非核苷类逆转录酶抑制剂（NNRTI），对1型人类免疫缺陷病毒（HIV-1）具有活性，且对目前已获许可的NNRTIs耐药。采用野生型病毒和NNRTI耐药病毒进行连续传代实验，以鉴定TMC125在体外选择的突变。除了在低感染复数（MOI）下使用递增浓度抑制剂进行的“经典”选择实验外，还进行了在高MOI下使用固定浓度抑制剂的实验，以确保TMC125与当前NNRTIs之间的标准化比较。低MOI和高MOI实验均表明，对TMC125产生耐药需要多个突变，这些突变常常导致对依非韦伦和奈韦拉平产生交叉耐药。在高MOI实验中，与依非韦伦和奈韦拉平不同，1μM TMC125可完全抑制野生型和NNRTI耐药HIV-1中耐药病毒的突破。此外，定点突变体（SDM）SDM-K103N/Y181C的病毒突破与野生型HIV-1在使用依非韦伦或奈韦拉平时的突破同时或更晚出现。选择实验鉴定出TMC125选择的突变，包括已知的与NNRTI相关的突变L100I、Y181C、G190E、M230L和Y318F，以及新突变V179I和V179F。检测TMC125对一组SDM的抗病毒活性表明，这些单个突变对耐药性的影响高度依赖于共存突变的存在和特性。这些结果表明，TMC125对NNRTI耐药病毒具有独特的活性谱，并且在体外对耐药性的产生具有较高的遗传屏障。
