PF-3450074 (PF-74) acts at an early stage of HIV-1 infection inhibits viral replication by directly competing with the binding of CPSF6 (nuclear host factors cleavage and polyadenylation specific factor 6) and NUP153 (nucleoporin 153), and blocks the uncoating, assembly, and the reverse transcription steps of the viral life cycle. PF-3450074 is a specifical inhibitor of HIV-1 capsid protein (CA) and shows a broad-spectrum inhibition of HIV isolates with submicromolar potency (EC50=8-640 nM).

HIV-1:0.72 μM(NL4.3 strain)

PF-3450074 displays a good potency in primary human peripheral blood mononuclear cells (PBMCs), inhibits HIV-193RW025, HIV-1JR-CSF and HIV-193MW965 with IC50 values of 1.5 ± 0.9 μM; 0.6 ± 0.20 μM; and 0.6 ± 0.10 μM, respectively. PF-3450074 shows anti-viral activities against HIV wild type NL4-3 and HIV T107N mutant (EC50: 0.72 μM and 4.5μM, respectively). PF-3450074 (10 μM; 8 hours) causes a marked reduction in late products of reverse transcription in HeLa-P4 cells with DNase I-treated stocks of Env-defective HIV-1 (R9.Env-). This compound shows Median IC50?and CC50?values of 0.9 ± 0.5 μM and 90.5 ± 5.9 μM, respectively. The KD?for the interaction between?PF-74?and the CA hexamer, derived in the same manner as for NUP153, is determined to be 176 ± 78 nM [1][2].

HeLa-P4 cells were inoculated with PF74 (10 μM). After 8 h of culture, the cells were harvested and DNA isolated with a DNeasy Blood & Tissue kit (Qiagen). HIV-1 DNA in the samples was quantified by real-time PCR using primers specific which shows that PF74 inhibited HIV-1 reverse transcription in target cells.