Evaluation of Anti-HIV-1 Compounds

The anti-HIV-1 activity of compounds 1–5 was evaluated using VSV-G pseudotyped NL43_GFP infection of human monocytic THP-1 cells. THP-1 and NL4–3_GFP were generously provided by Lionel Berthoux and Amita Singh and are described in Ka et al. (2021) [13 (link)]. THP-1 cells were seeded at 2.0 × 10⁴ cells per well in 96 well-plates. The next day, cells were treated with 4 concentrations of each compound (12.5, 25, 50 and 100 mM) and then infected with HIV_GFP at a MOI of 1. After 72 h, cells were stained with propidium iodide (PI, 0.5 mg/mL) and both PI⁺ and HIV-1_GFP⁺ infected cells frequencies were assessed on a FC500 MPL cytometer (Beckman Coulter, Inc., Mississauga, ON, Canada) and analyzed using FlowJo software (FlowJo LLC, BD Biosciences, Ashland, OR, USA). Matched concentrations of dimethyl sulfoxide (DMSO) were used as negative controls. All infection assays were performed in triplicate.