A-443654 is a pan-Akt inhibitor. A-443654 has equal potency against Akt1, Akt2, or Akt3 within cells (Ki=160 pM).

A-443654 demonstrates a significant improvement in potency with a K_i of 160 pM, a 30,000-fold increase compared to the initial lead molecule. It effectively reduces P-GSK3 in a dose-dependent manner across three cell lines and exhibits 40-fold greater selectivity for Akt over PKA, equally inhibiting Akt1, Akt2, and Akt3 within cells. Morphological changes induced by A-443654 are rapid, occurring within 2 to 4 hours in 10A and 10CA1a cells, with the latter being more sensitive. The compound hinders tumor cell proliferation, with an EC50 of 0.1 μM[1], and shows marked selectivity for mutant cells, inhibiting their growth more than 3.5 times compared to wild-type (WT) cells[3]. Furthermore, A-443654, especially when combined with rapamycin, significantly decreases Bcl-2 protein levels in 10A and 10CA1a cells, exhibiting a notable reduction in 10CA1a cells by approximately 70%. At 2 μM, A-443654 alone prompts 10CA1a cells to detach from the plate after 12 hours, illustrating a differential response compared to 10A cells, which remain unaffected at the same concentration[2].

A-443654, administered subcutaneously at a dosage of 7.5 mg/kg per day, effectively inhibits tumor growth in the 3T3-Akt1 flank tumor model. At a higher concentration of 30 mg/kg, this compound increases phosphorylated Akt1 levels in MiaPaCa-2 tumors. Furthermore, a dosage of 50 mg/kg induces apoptosis in 3T3-Akt1 flank tumors [1].