Abstract 732: Targeting the Bcl-2/BH3 protein-protein interaction: Identification, design, and synthesis of new selective inhibitors of the Mcl-1/BH3 interaction

It is well accepted that escape of apoptosis is a major mechanism for cancer progression and drug resistance. Anti-apoptotic members of the Bcl-2 protein family, such as Bcl-2, Bcl-x_L and Mcl-1, are among the most effective inhibitors of apoptosis and are frequently upregulated in various types of human cancer. A number of pro-apoptotic proteins share only the BH3 domain with other members of the Bcl-2 family, so called BH3-only proteins, and appear to function essentially as transdominant inhibitors by binding to anti-apoptotic Bcl-2 family proteins and neutralizing their cell-survival activity. The development of small molecules to mimic the BH3 proteins has been an intense area of research. It is clear from the development of ABT737 (and its orally available relative ABT-263) that targeting of Bcl-2/Bcl-x_L by small molecule BH3-mimetics is a promising strategy for the development of a new class of anticancer drugs. However the clinical potential of ABT737 may be limited since it does not inhibit the pro-survival Bcl-2 family protein Mcl-1. To date available Bcl-2 family proteins inhibitors are either pan-inhibitors such as (-)-gossypol (which simultaneously inhibits Bcl-xl/BH3, Bcl-2/BH3 and Mcl-1/BH3 interactions) or are selective for the Bcl-xl/BH3 and Bcl-2/BH3 interactions. Hence a selective Mcl-1/BH3 inhibitor would greatly aid investigating the role of Mcl-1 in cancer and in drug resistance and complement the activity of ABT737. Our current study is centered on the identification and the synthesis of small molecule selective inhibitors of the Mcl-1/BH3 interaction. To this aim we interrogated the hit-list from inhibitors of the MLSCN screen of the Mcl-1-NOXA interaction [(Emory, AID 1022) TR-FRET screen for Mcl-1-NOXA inhibitors]. We docked 2100 of the experimental 2100 MLSCN hits with Mcl-1 protein structure derived from the Mcl-1-NOXA X-ray crystal structure. We then examined the top 375 compounds. We then ranked them by docking score, the number of MLSCN biological assays tested and number of hits in these assays. We then selected compounds that docked well and had been tested in many MLSCN assays for which they were not hits. Selected compounds were purchased and assessed for activity in an Alphascreen and ELISA assay, to reveal the hit HL5-026 (IC₅₀ = 46 µM). HL5-026 is an inhibitor of the Mcl-1/Bim-BH3 interaction that has four-fold selectivity over Bcl-xl/Bim-BH3. HL5-026 binding to Mcl-1 is currently being assessed via co-crystallization and other biophysical techniques. We will report the synthesis and biological evaluation of focused libraries based on the initial hit and designed by modeling and docking to Mcl-1. First generation libraries have revealed compounds of improved activity. Structure activity relationships around the hits will be disclosed as well as the outcomes of further rounds of chemical design and biological assessment.

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 732.