Abstract 5701: AU2-94: A CDK4-specific inhibitor with the marked anti-tumor efficacy

Abstract Cyclin-dependent kinases (CDKs) are a vital family of proteins in the cell development. Several CDK4/6 inhibitor drugs significantly extended the lives of subtypes breast cancer patients. However, they have some shortcomings. Many patients do not respond to them well mainly due to off-target toxicity requiring dose reductions and treatment interruptions, and thus building resistance, which limit their effectiveness. Here, we describe AU2-94, a highly selective CDK4 inhibitor, for the treatment of CDK4-dependent tumours including breast cancer (ER+ and TNBC), colorectal cancer and glioblastoma (GBM). AU2-94 exhibits high selectivity for CDK4 (Ki = 2 nM) over CDK6 (Ki = 279 nM). Consistent with the CDK4-targeted mechanism, AU2-94 arrested the G1 cells and inhibited cancer cell proliferation. Interestingly, AU2-94 reduced cellular CDK2, Cyclin E2, Cyclin A2, Cyclin B1, and induced apoptosis via reduction of Bcl-2, FOXM1 etc. In contrast to cancer cells, AU2-94 has little impact on the cell cycle, senescence, or apoptosis of human bone marrow cells, whereas the known CDK4/6 inhibitors increased the G1 cells. AU2-94 showed little or no effect on lymphocyte and neutrophil counts whilst the clinical CDK4/6 inhibitors reduced them. This lower effect of AU2-94 on the bone marrow, neutrophils and lymphocytes might be due to its lesser effect on CDK6 compared to the clinical CDK4/6 inhibitors. AU2-94 was highly efficacious against multiple in vivo tumour models. In the ER+ breast cancer T47D xenografts, AU2-94 caused tumour regression. In the model of MDA-MB-453 AR+ luminal breast cancer (CDK4, KRASmut, PI3KCAmut), AU2-94 demonstrated remarkable efficacy, resulting in the 6 out of 7 (86%) mice tumour-free. In the colorectal cancer HCT116 (CDK4/6, KRASmut) xenografts, AU2-94 inhibited tumour growth markedly in a dose-dependent manner from 25 to 200 mg/kg without any overt toxicity. The pharmacokinetic and pharmacodynamic study revealed up to 500-fold higher concentration of AU2-94 in the tumours than in the plasma, suggesting its tumour targeting specificity. Several robust biomarkers/PD markers were identified. The Rb-CDK4-Cyclin D pathway is altered in the majority of GBM due to homozygous deletion of CDKN2A/B. AU2-94 has an excellent therapeutic potential as it crossed the blood-brain-barrier effectively. In the both U87 subcutaneous and orthotopic models, AU2-94 reduced the tumour burden and increased the survival of GBM bearing mice significantly compared to vehicle. Moreover, AU2-94 synergised the anti-cancer efficacy of PI3K/Akt/mTOR and HER2 inhibitors in combination. In summary, AU2-94 is a CDK4 inhibitor with high selectivity against a panel of 360 human kinases. This selectivity can be a reason for the better safety of AU2-94 compared to the clinical CDK4/6 inhibitors. The high oral bioavailability, robust antitumor efficacy and excellent safety profiles make AU2-94 a highly attractive candidate for clinical development. Citation Format: Laychiluh Bantie, Jimma Likisa, Muhammed Rahaman, Theodosia Teo, Ava Safaroghliazar, Sunita KC Basnet, Ramin Hassankhani, Benjamin Noll, Robert Milne, Shudong Wang. AU2-94: A CDK4-specific inhibitor with the marked anti-tumor efficacy abstract. In: Proceedings of the American Association for Cancer Research Annual Meeting 2024; Part 1 (Regular Abstracts) ; 2024 Apr 5-10; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2024;84 (6Suppl): Abstract nr 5701.