Anti-HIV Agents ; chemistry ; pharmacology ; Benzoxazines ; Coumarins ; chemistry ; pharmacology ; Delavirdine ; chemistry ; pharmacology ; Drug Resistance, Viral ; HIV Reverse Transcriptase ; drug effects ; HIV-1 ; drug effects ; Humans ; Molecular Conformation ; Molecular Structure ; Nevirapine ; chemistry ; pharmacology ; Oxazines ; chemistry ; pharmacology ; Pyranocoumarins ; Reverse Transcriptase Inhibitors ; chemistry ; pharmacology

Consistent non-nucleoside reverse-transcriptase inhibitors (NNRTIs) resistant HIV-1 strains occurred due to the clinical use for more than ten years of efavirenz (EFV), nevirapine (NVP), and delavirdine (DLV). In this study, we established nine cell-based pharmacological models according to most NNRTIs-resistant clinical tested strains, Resistant mutations were introduced into vector, pNL4-3.Luc.R-E-, by overlapping PCR. Then, pseudovirions were produced by co-transfection of VSV-G plasmid and pNL4-3.Luc.R-E- -mut. All nine recombinant VSVG/HIV-mut pseudovirions (VSVG/HIV-wt, VSVG/HIV(-K103N), VSVG/HIV(-Y181C), VSVG/HIV(-L100I,K103N), VSVG/HIV(-Y188L), VSVG/HIV(-K103N,Y181C), VSVG/HIV(-K103N,P225H), VSVG/HIV(-K103N,Y188L), VSVG/HIV(-K103N,G109A) and VSVG/HIV(-K103N,V108I)) had high efficient infectivity. Furthermore, they all showed resistant characteristics to EFV and NVP with IC50 changes consisting with clinical reports, not to nucleoside reverse-transcriptase inhibitors (AZT and d4T). This series safe cell-based model, which could be carried out in BSL-2 laboratory, can be used for evaluating NNRTIs candidates.
Anti-HIV Agents ; pharmacology ; Benzoxazines ; pharmacology ; Cell Line ; Delavirdine ; pharmacology ; Drug Evaluation, Preclinical ; methods ; Drug Resistance, Viral ; Genetic Vectors ; HIV Reverse Transcriptase ; antagonists & inhibitors ; genetics ; metabolism ; HIV-1 ; drug effects ; genetics ; Humans ; Membrane Glycoproteins ; genetics ; Nevirapine ; pharmacology ; Plasmids ; genetics ; Point Mutation ; Reverse Transcriptase Inhibitors ; pharmacology ; Stavudine ; pharmacology ; Transfection ; Viral Envelope Proteins ; genetics ; Virion ; genetics ; metabolism ; Virus Replication ; Zidovudine ; pharmacology