Synthesis and HIV-1 Inhibition Assays

Compounds 13 and 21 were synthesized as previously reported [30 (link)]. The inhibitors: azidothymidine (AZT); efavirenz (EFV); nevirapine (NVP) and raltegravir (RAL) were purchased form Sigma-Aldrich (St. Louis, MO, USA). RNase H inhibitor was kindly provided by Prof. Roberto Di Santo. MT4 (ECACC 08081402) and HEK 293 T cells (ATCC® CRL-3216TM) were cultured in RPMI 1640 medium and Dulbecco’s modified Eagle’s medium (DMEM) (Gibco, Waltham, MA, USA), respectively. Both media were supplemented with 10% fetal calf serum (FCS) (Gibco, Waltham, MA, USA) and 1% penicillin/streptomycin. Stocks of HIV-1 NL4-3 strain were prepared by transfecting 293 T cells with the HIV-1 pNL4-3. The VSV-pseudotyped, NL4-3–GFP expressing virus (VSV/HIV-1_GFP) was generated as reported [32 (link)].