Vingerhoets J, Azijn H, Fransen E, De Baere I, Smeulders L, Jochmans D, et al. TMC125 displays a high genetic barrier to the development of resistance: evidence from in vitro selection experiments

Tibotec, Mechelen, Belgium.
Journal of Virology (Impact Factor: 4.65). 11/2005; 79(20):12773-82. DOI: 10.1128/JVI.79.20.12773-12782.2005
Source: PubMed

ABSTRACT 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.

Download full-text


Available from: Dirk Jochmans, Aug 17, 2015
  • Source
    • "However, based on the weighted mutation score algorithm for the prediction of ETR susceptibility [Vingerhoets et al., 2010b], some of the HIV-1 isolated strains (23.6%) had mutations or combinations of mutations associated with intermediate resistance to ETR. Furthermore, L100I was the most prevalent ETR RAM found (16.4%) and in combination with K103N, a first-generation NNRTIs RAM widely present in EFV resistant viruses [Tambuyzer et al., 2009], was described to be associated in vitro with intermediate resistance to ETR [Vingerhoets et al., 2005]. Therefore, careful genotypic analysis is necessary when the use of ETR is considered. "
    [Show abstract] [Hide abstract]
    ABSTRACT: Etravirine (ETR) is a non-nucleoside analogue reverse transcriptase inhibitor (NNRTI) with a high genetic barrier to the development of resistance and with potential activity against Human immunodeficiency virus type 1 (HIV-1) strains resistant to first-generation NNRTIs. The objective of this study was to investigate the prevalence of ETR resistance associated mutations (RAMs) in HIV-1 strains isolated from infected individuals failing efavirenz (EFV), as well as to evaluate possible differences in the distribution of ETR RAMs between subtype B and non-B genetic variants. Nucleotide sequences of the protease and partial reverse transcriptase (RT) coding regions of the pol gene of 55 HIV-1 strains isolated from infected individuals failing EFV on regular follow-up at a reference center in Portugal, were retrospectively analyzed. The most prevalent ETR RAMs observed were L100I, V90I, and K101E, with a prevalence of 16.4% (n = 9), 9.1% (n = 5), and 5.5% (n = 3), respectively. Overall, 47.3% (n = 26) of the nucleotide sequences had at least one ETR RAM: 38.2% (n = 21) had one ETR RAM, 7.3% (n = 4) had two ETR RAMs and 1.8% (n = 1) had three ETR RAMs. No statistically significant differences were found in the distribution of ETR RAMs between subtype B and non-B genetic variants. The results demonstrate that ETR rescue therapy is a viable option in treatment-experienced individuals failing EFV and suggests that ETR may be equally useful in HIV-1 infections caused by different genetic variants.
    Journal of Medical Virology 04/2012; 84(4):551-4. DOI:10.1002/jmv.23232 · 2.22 Impact Factor
  • Source
    • "4-[6-Amino-5-bromo-2-[(4-cyanophenyl)amino] pyrimidin-4-yl]oxy-3,5- dimethylbenzonitrile (etravirine; ETR) is a second-generation NNRTI that, like other NNRTIs, functions by binding noncompetitively to HIV reverse transcriptase, thereby preventing synthesis of DNA from the RNA genome of the virus. As a diarylpyrimidine, ETR has a high capacity for isomerization, which allows it to effectively bind to and inhibit common mutated forms of the viral enzyme (Andries et al., 2004; Vingerhoets et al., 2005; Gupta et al., This work was supported by the PhRMA Foundation (Research Starter Grant in Pharmacology/Toxicology; to N.N.B.). The AB SCIEX QTRAP 5500 mass spectrometer was purchased using a grant from the National Institutes of Health National Center for Research Resources [Grant 1S10-RR27733]; the Waters Acquity ultraperformance liquid chromatograph interfaced with the AB SCIEX QTRAP 5500 mass spectrometer was purchased with funds provided by the Pendleton Foundation Trust. "
    [Show abstract] [Hide abstract]
    ABSTRACT: Etravirine (ETR) is a second-generation non-nucleoside reverse transcriptase inhibitor prescribed for the treatment of HIV-1. By using human liver microsomes (HLMs), cDNA-expressed cytochromes P450 (P450s), and UDP-glucuronosyltransferases (UGTs), the routes of ETR metabolism were defined. Incubations with cDNA-expressed P450 isozymes and chemical inhibition studies using HLMs indicated that CYP2C19 is primarily responsible for the formation of both the major monohydroxylated and dihydroxylated metabolites of ETR. Tandem mass spectrometry suggested that these metabolites were produced via monomethylhydroxylation and dimethylhydroxylation of the dimethylbenzonitrile moiety. Formation of these monohydroxy and dihydroxy metabolites was decreased by 75 and 100%, respectively, in assays performed using HLMs that were genotyped as homozygous for the loss-of-function CYP2C19*2 allele compared with formation by HLMs genotyped as CYP2C19*1/*1. Two monohydroxylated metabolites of lower abundance were formed by CYP3A4, and interestingly, although CYP2C9 showed no activity toward the parent compound, this enzyme appeared to act in concert with CYP3A4 to form two minor dihydroxylated products of ETR. UGT1A3 and UGT1A8 were demonstrated to glucuronidate a CYP3A4-dependent monohydroxylated product. In addition, treatment of primary human hepatocytes with ETR resulted in 3.2-, 5.2-, 11.8-, and 17.9-fold increases in CYP3A4 mRNA levels 6, 12, 24, and 72 h after treatment. The presence of the pregnane X receptor antagonist sulforaphane blocked the ETR-mediated increase in CYP3A4 mRNA expression. Taken together, these data suggest that ETR and ETR metabolites are substrates of CYP2C19, CYP3A4, CYP2C9, UGT1A3, and UGT1A8 and that ETR is a PXR-dependent modulator of CYP3A4 mRNA levels.
    Drug metabolism and disposition: the biological fate of chemicals 01/2012; 40(4):803-14. DOI:10.1124/dmd.111.044404 · 3.33 Impact Factor
  • Source
    • "ETV resistance profile was further evaluated by testing the molecule against thousands of NNRTI resistant HIV-1 clinical strains (Rimsky et al., 2009). In vitro, ETV shows a higher genetic barrier to the development of resistance when compared to NVP and EFV (Vingerhoets et al., 2005). ETV- RT crystallographic complexes with the K103N mutation allowed for a better understanding of the molecule binding mode to the enzyme. "
    [Show abstract] [Hide abstract]
    ABSTRACT: In recent years relevant progress has been made in the treatment of HIV-1 with a consequent decrease in mortality. The availability of potent antiretroviral drugs and the ability of viral load assays that accurately evaluate the true level of viral replication, have led to a better understanding of pathogenesis of the disease and how to obtain improved therapeutic profiles. The highly active antiretroviral therapy (HAART), based on a combination of three or more antiretroviral drugs, has radically changed the clinical outcome of HIV. In particular, reverse transcriptase non-nucleoside inhibitors (NNRTIs) play an essential role in most protocols and are often used in first line treatment. The high specificity of these inhibitors towards HIV-1 has increased the number of structural and molecular modeling studies of enzyme complexes and that have led to chemical syntheses of more selective second and third-generation NNRTIs. However, a considerable percentage of new HIV-1 infections are caused by the emergence of drug-resistant mutant viruses that complicate treatment strategies. In this review we discuss relevant clinical and structural aspects for the management of antiretroviral drug resistance, with detailed explanations of mechanisms and mutation patterns useful to better understand the relation between drug resistance and therapy failure.
    Drug resistance updates: reviews and commentaries in antimicrobial and anticancer chemotherapy 02/2011; 14(3):141-9. DOI:10.1016/j.drup.2011.01.002 · 8.82 Impact Factor
Show more