Abstract P3-04-15: The PI3K-inhibitor, copanlisib, has selective activity in luminal breast cancer cell lines and shows robust combined activity with hormonal blockade and CDK-4/6 inhibition in ER+ breast cancer cell line xenografts
Authors: O'Brien NA , Conklin D , Luo T , Ayala R , Issakhanian S , Kalous O , Von Euw E , Politz O , Wilhelm S , Childs BH , Hurvitz SA , Slamon DJ 
Published: May 2019

Abstract
Background: Genetic and epigenetic alterations in the PI3K/mTOR and cyclin D:CDK-4/6:Rb signaling axes occur frequently in breast cancer and have been attributed to resistance to both ER- and HER2-directed therapeutics. Pharmacologically targeting CDK-4/6 in combination with hormonal blockade provides clinical benefit in patients with advanced ER+ breast cancer. In this study, we evaluated the activity of the pan-class I PI3K inhibitor, copanlisib (BAY-80-6946), with potent alpha and delta activity as a single agent or in combination with CDK-4/6 inhibition and hormonal blockade in a panel of breast cancer cell lines.
Methods: The growth inhibitory activity of copanlisib was evaluated against a large panel of 48 breast cancer cell lines molecularly characterized by genomic, transcriptomic and proteomic profiling. IC50 values were determined from direct cell counts using a Z1-particle counter. The activity of copanlisib in combination with hormone blockade and CDK-4/6 inhibition, by palbociclib, was assessed in two cell line xenograft models of ER+ breast cancer; MCF7(PIK3CA-E545K) and ZR751(PIK3CA WT). For xenograft studies, tumor bearing mice were treated once weekly (BID) by intravenous injection with clinically achievable doses of copanlisib (10 mg/kg) as single agent or in combination with tamoxifen or fulvestrant with or without 75 mg/kg daily palbociclib for 21 days.
Results: A broad range of IC50 values (0.491-895 nM), with a high degree of separation between sensitive and resistant histologically defined subgroups were determined for copanlisib, indicating the potential for a wide therapeutic window. Luminal subtype, the presence of activating mutations in PIK3CA, high levels of ER, HER2, HER3 and EGFR protein enriched for sensitivity to copanlisib. Activating mutations of KRAS and BRAF were associated with resistance to copanlisib. Single agent copanlisib induced significant tumor growth inhibition (TGI) relative to vehicle control in each of the xenograft models. Modest increases in anti-tumor activity were achieved when copanlisib was combined with hormonal blockade by either tamoxifen or fulvestrant. However, robust tumor regressions were observed with the triple combinations of copanlisib-palbociclib-tamoxifen and copanlisib-palbociclib-fulvestrant. Furthermore, these triple combinations achieved a statistically significant improvement in anti-tumor activity over the standard of care combination of palbociclib plus fulvestrant. Each of the single agent and treatment combinations tested were well tolerated in animals.
Discussion: These preclinical data illustrate the potent and selective activity of the pan class I PI3K inhibitor copanlisib in luminal breast cancers and support the clinical investigation of copanlisib in combination with CDK-4/6 inhibition and hormonal blockade in ER+ breast cancer.
Citation Format: O'Brien NA, Conklin D, Luo T, Ayala R, Issakhanian S, Kalous O, Von Euw E, Politz O, Wilhelm S, Childs BH, Hurvitz SA, Slamon DJ. The PI3K-inhibitor, copanlisib, has selective activity in luminal breast cancer cell lines and shows robust combined activity with hormonal blockade and CDK-4/6 inhibition in ER+ breast cancer cell line xenografts [abstract]. In: Proceedings of the 2016 San Antonio Breast Cancer Symposium; 2016 Dec 6-10; San Antonio, TX. Philadelphia (PA): AACR; Cancer Res 2017;77(4 Suppl):Abstract nr P3-04-15.