In prior investigation, we discovered that (3′R,4′R)-3-cyanomethyl-4-methyl-3′,4′-di-O-(S)-camphanoyl-(+)-cis-khellactone (4, 3-cyanomethyl-4-methyl-DCK) showed promising anti-HIV activity. In these current studies, we developed and optimized successfully a practical 10-step synthesis for scale-up preparation to increase the overall yield of 4from 7.8% to 32%. Furthermore, compound 4exhibited broad-spectrum anti-HIV activity against wild-type and drug-resistant viral infection of CD4+ T cell lines as well as peripheral blood mononuclear cells by both laboratory-adapted and primary HIV-1 isolates with distinct subtypes and tropisms. Compound 4was further subjected to in vitro and in vivo pharmacokinetic studies. These studies indicated that 4has moderate cell permeability, moderate oral bioavailability, and low systemic clearance. These results suggest that 4should be developed as a promising anti-HIV agent for development as a clinical trial candidate. [ABSTRACT FROM AUTHOR]