The Third-generation Anti-hiv Drug Screening Platform Establishment And Applications

Current clinical control of HIV-1 infection is the triple drug regimens that comprise combination of reverse transcriptase and protease inhibitors which called the first generation of anti-HIV drugs. Qwing to intolerance and virus resistance during the long-term therapy, it is necessary to modify the inhibitors of reverse transcriptase and protease and development the second generation of anti-HIV drugs. Because of the persistence of infection in reservoirs with low turnover rates and the limitation of the first and second generation anti-HIV drugs, eradication of the infected HIV appears unlikely. As a consequence, the search for new anti-HIV agents renewed emphasis on HIV entry, which called the third generation of anti-HIV and have advantages over the past in intolerance and side effect because of the different mechanism. Combination of the three generations of anti-HIV drugs will improve the effect of clinical therapy.Cyclophilin A (CYPA) is specifically incorporated into the virions of HIV-1 and plays an essential role in virus morphogenesis through a specific interaction with the gag precursor polyprotein, and the incorporated CYPA has shown to enhance significantly an early step of cellular HIV-1 infection by CYPA-CD147 interaction. Two anti-HIV drug screening models are constructed based on the interaction of CYPA-CD147 and CYPA-Gag, which can screen and identify the active compounds in combinant chemical library and in natural products extacts from herbs or microbial metabolites. The HIV envelope glycoprotein gp41 plays an important role in the fusion of viral and target cell membranes and serves as an attractive target for development of HIV fusion inhibitors. The peptides libraries derived from N- and C-terminal heptad repeats (NHR and CHR) of gp41 are constructed and screened the intensely binding pairs by BacteriaMatch system. After NHR and CHR fusion with GFP and Rluc gene respectively, the fusion genes clone into pET22b and pET28a and express in BL21(λ DE3) respectively, the expression products can screen the compound libraries directly or after purification by BRET. These screening systems can not only identify the identify the active compounds in combinant chemical library and in natural products extacts from herbs or microbial metabolites, but also evaluate the activities of NHR and CHR mutants.