Warangalone is an anti-malarial compound that inhibits the growth of Plasmodium falciparum strains 3D7 (chloroquine-sensitive) and K1 (chloroquine-resistant) with IC50 values of 4.8 μg/mL and 3.7 μg/mL, respectively.

Parasite (K1):3.7 μg/mL, CAK:3.5 μM, Parasite (3D7):.8 μg/mL

Warangalone can also inhibit cyclic AMP-dependent protein kinase catalytic subunit (cAK) with an IC50 of 3.5 μM [2]. When HL-60 cells are exposed to Warangalone (30 μM) for 24 h, Warangalone induces a significant decrease (8%) in cell viability compared to controls. Warangalone also inhibits HL-60 cell growth within 24 h in a time-dependent fashion. A time-dependent increase in caspase-9 activity is observed in Warangalone-treated cells [3].

Cells are seeded in 24-well plates at a density of 3×10^6 cells per well. After exposure of the cells to Warangalone for the allotted time periods, the cells are washed three times with PBS and then lysed in a lysis buffer for 10 min on ice. The protein content of the cell lysates is assayed with a Micro BCA reagent. Cell lysates containing 50 μg of protein are incubated with a caspase-3 fluorogenic substrate (DEVD-AFC) or a caspase-9 fluorogenic substrate (LEHD-AFC) for 1 h at 37°C. Caspase activity is measured by fluorometric detection [3].

Cells are seeded in 96-well plates at a density of 1×10^5 cells per well. The cells are maintained for 24 h at 37°C and then Warangalone (30 μM) is added to the culture medium. MTS solution is added to the 96-well plates at the indicated time points, and the cells are incubated for 1 h at 37°C. The absorbance is measured at a wavelength of 490 nm with a microplate counter [3].