Abstract LB-116: GSK2334470: A potent and highly selective inhibitor of PDK1

Abstract

Phosphoinositide-dependent protein kinase-1 (PDK1) is a master regulator of the AGC family of kinases and an integral component of the PI3K/AKT/mTOR pathway which is the most commonly deregulated signaling pathway across all cancers. In vivo studies have shown that AKT, p70S6K, RSK and protein kinase C are key mediators of PDK1 function, regulating diverse cellular processes. Activation of these substrates by PDK1 leads to an increase in glucose uptake, protein synthesis, and inhibition of pro-apoptotic proteins. Conversely, knockdown of PDK1 protein levels in tumor cells leads to decreased tumor cell proliferation and increased apoptosis. Furthermore, a hypomorphic mutation of PDK1 suppresses tumorigenesis in PTEN^+/- mice. Taken together, these observations suggest that an inhibitor of PDK1 could be beneficial in treating cancer. Therefore, we initiated a medicinal chemistry program to discover PDK1 inhibitors as potential anticancer agents.

We have identified an aminoindazole PDK1 inhibitor, GSK2334470, derived from a fragment screening hit which we optimized using structure-based design and PDK1 protein crystallography. Potency and kinase selectivity were attributed to key interactions within the ATP binding site between the inhibitor and the back pocket and glycine-rich loop of PDK1. GSK2334470 is a potent inhibitor of PDK1 kinase (IC₅₀ = 0.5 nM) with a high level of specificity for PDK1 based on test results from in an in vitro kinase selectivity panel of more than 280 kinases. In PC-3 cells, GSK2334470 effectively inhibited phosphorylation of AKT^T308 (IC₅₀ = 113 nM) and RSK^S221 (IC₅₀ = 293 nM) but not AKT^S473 (IC₅₀ > 30,000 nM). These results are consistent with the high specificity of GSK2334470 for PDK1 inhibition. In a panel of approximately 300 cell lines, GSK2334470 demonstrated modest antiproliferative activity overall, with low micromolar activity seen in several breast cancer cell lines and in a variety of hematological cancer cells. Interestingly, GSK2334470 displayed sub-micromolar antiproliferative activity against AML cell lines that are clinically classified as M4 and M5 FAB-subtypes. Therefore, we evaluated the pharmacodynamics of GSK2334470 in mice implanted with OCI-AML2 xenografts. GSK2334470 dosed i.p. at 100 mg/kg resulted in 58 and 29% inhibition of AKT^T308 phosphorylation at 3 and 6 h, respectively and 57 and 71% inhibition of RSK^S221 phosphorylation at 3 and 6 h, respectively. There was no observed change on AKT^S473 phosphorylation. Our results demonstrate that GSK23334470 is a potent, highly selective PDK1 inhibitor which can decrease PDK1 signaling in a tumor xenograft model.

Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 101st Annual Meeting of the American Association for Cancer Research; 2010 Apr 17-21; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2010;70(8 Suppl):Abstract nr LB-116.