Targeting Akt as strategy to kill cancer cells using 3-substituted 5-anilinobenzo[c]isoxazolequinones: A preliminary study

Several new 3-substituted 5-anilinobenzo[c]isoxazolequinones were synthesized from 1,4-benzoquinone and alkyl-or arylcarbaldehydes by a three-step synthetic sequence. The new compounds (3a-h) were tested in vitro in normal human fibroblasts and two cancer cell lines for their cytotoxic activity. The range of IC50 values obtained for the compounds was from 3.4 to 74.2 mu M. Five members of the series (3b, 3d, 3e, 3f, 3g) were further selected and evaluated as inhibitors of the Hsp90 chaperoning function taking Akt as example of Hsp90 client proteins. We also evaluated the changes of intracellular levels of GSH and ATP as markers of cellular metabolic status in response to these compounds in T24 cells. One of such isoxazolquinones (3b) decreased the expression of Akt, PARP and Hsp90. Compounds 3b and 3d decreased the amount of ATP but caused no effect on GSH levels. These compounds also activated caspase-3 but an apoptosis-like type of cell death was unlike since PARP protein was not cleaved and caspase activation was substantially lower than its activation induced by staurosporine, a known caspase-3 activator in T24 cells. Taken together, preliminary results led to the discovery of an original lead compound (3b) which can be used as model to obtain new Akt inhibitors.