Spectinamide-1599 Synergistic Effects

Spectinamide-1599 was synthesized as described (Lee et al., 2014 (link)) and provided by Dr. Richard Lee (St. Jude Children’s Research Hospital). Pyrazinamide, spectinomycin and streptomycin were purchased from Sigma-Aldrich (Millipore Sigma, St. Louis, MO, United States) and recombinant murine Interferon Gamma (IFN-γ) from PeproTech (Rocky Hill, NJ, United States). Stock solutions for each drug were prepared as follow; Spectinamide-1599 was diluted at 300 μg/mL with cRPMI Medium. Pyrazinamide was dissolved in cRPMI medium at 20 mg/mL and IFN-γ was dissolved in cRPMI medium at 300 U/mL. On day 2 post-infection cell cultures received drug treatment. Cell cultures were treated with monotherapy or combinations of 200 or 400 μg/mL of PZA, 25 or 100 μg/mL spectinamide-1599 and 50 U/mL murine IFN-γ.