Hiv-1 Reverse Transcriptase Inhibitors Jl2, Ephcl, And Ebzq Role Of Targets Identified And Mechanisms

Human Immunodeficiency Virus(HIV)is a retrovirus,belongs to Lentiviridae family.As long as viral genetic material entering into host cytoplasm,double-strand DNAs synthesis occurs which is catalyzed by reverse transcriptase(RT)with viral plus-strand RNA as template.The process of reverse transcription can be divided into several steps:formation of minus-strand strong-stop DNA;the first translocation; initiation of plus-strand DNA synthesis;the second translocation and the completion of both strands.We applied Polymerase Chain Reaction(PCR)method to detect the effect of compounds JL2,EPHCL and EBZQ on the inhibition of four steps in the process of HIV-1 reverse transcription.According to the amplified products on the region of R/U5,U3,U5/PBS and the sequence between LTR and Gag,we judged the effect of the three compounds on the formation of minus-strand DNA,the first translocation,the initiation of plus-strand DNA synthesis,and the completion of both strands.VSVG/MLV-GFP model was used to detect the specificity of the three compounds on the process of reverse transcription under high concentration(10^-5M), which showed all three compounds have no inhibition on the replication of MLV. Results of the experiments showed that compounds JL2,EPHCL and EBZQ could inhibit the HIV-1 reverse transcription with specificity under the 10-fold of 50% inhibiting concentration.They had no significant effect on the initiation of reverse transcription,and could restrain the synthesis of full-length double-stranded DNA. These indicate that compounds JL2,EPHCL and EBZQ inhibit HIV-1 replication through affecting the process of reverse transcription,and the key site is during the processes of the second translocation and the elongation of the minus- and plus-strand before synthesizing the full-length double-strand DNA.