Abstract A104: AZD1390, a potent and selective orally bioavailable blood-brain barrier-penetrant ATM inhibitor, radiosensitizes and improves survival of orthotopic glioma and metastatic brain tumor models
Authors: Durant Steve T. , Pike Kurt G. , Colclough Nicola , Riches Lucy , Garcia-Trinidad Antonio , Hunt Thomas , Ling Stephanie , Stott Jonathan , Barrett Ian , Zheng Li , Wang Yingchun , Chen Kan , Zhang Tianwei , Pilla Reddy Venkatesh , Sykes Andrew , Johnstrom Peter , Jones Gemma , Pierce Andrew , Karlin Jeremy , Kahn Jenna , Allen Jasmine , Valerie Kristoffer , Illingworth Ruth , Pass Martin 
Published: August 2018

Abstract
ATM plays a central role in the detection, signalling, and repair of DNA double-strand breaks (DSB), the most cytotoxic lesion induced by ionizing radiation (IR) and certain chemotherapies. ATM is also activated by reactive oxygen species (ROS) induced by cellular exposures to IR. Genetic ablation and pharmacologic inhibition of ATM is associated with extreme hypersensitivity of glioblastoma multiforme (GBM) tumor cells to IR, especially those with checkpoint defects such as p53 abrogations. GBM is the most common and lethal form of brain tumor. Median survival of patients is 12-15 months, despite surgery, fractionated radiotherapy, and temozolomide standard of care. Poor survival is attributed to an inability to excise all tumor tissue (if operable), dissemination of disease into regions with an intact blood-brain barrier (BBB), and an intrinsic radio- and chemo-resistance. With ATM activity robustly upregulated in GBM stem cells, ATM represents an attractive radiosensitization target. Here, we describe the activity of AZD1390, a potent, selective, and orally bioavailable ATM inhibitor optimized for BBB penetration in preclinical model species. AZD1390 demonstrates exquisitely potent cellular inhibition of ATM activity (IC50 0.78 nM) with &gt;1000-fold activity over closely related (PIKKs) and distant kinases. We confirm target and pathway engagement by Western blot and imaging pan-nuclear and discreet pATM foci staining (IC50 0.6-3nM). Radiosensitization of a panel of GBM cell lines and NCI-H2228 lung cells was confirmed in antiproliferation and clonogenic assays (IC50 3 nM). DEF37 of 2.7 was seen in p53 mutant GBM cells dosed at 3nM and p53 mutant GBM cell lines were more radiosensitized than wild type cells. Radiosensitization was confirmed in vivo in mouse orthotopic NCI-H2228 lung tumor models implanted directly into brain or via carotid artery injection showing dose-dependent tumor growth inhibition and remarkable increases in survival of mice when dosing AZD1390 PO an hour before four daily fractions of 2.5 Gy IR to the whole head. 20 mg/kg QD or BID gave the best survival benefit that correlated with bioluminescent tumor growth inhibition. Doses lower than 2 mg/kg were not efficacious, suggesting free brain PK cover over ATM IC50 of 3 hours or more are required for efficacy. Efficacy was also achieved in a dose-dependent manner in orthotopic GL261 murine GBM syngeneic models dosed in combination with either whole head radiotherapy or stereotactic beam radiotherapy. A PK PD efficacy relationship was establish by correlating AZD1390 free brain PK levels, phospho-ATM/Rad50 detection in tumor by IHC, and tumor growth inhibition and survival. Significant brain exposure was observed in a nonhuman primate PET study utilizing 11C-labelled AZD1390, further supporting the ability of the compound to efficiently cross the BBB. With confirmation that AZD1390 is not a substrate for human efflux transporters and having favorable pharmacokinetic and pharmacodynamic properties, AZD1390 is now in early clinical development for use as a radiosensitizer in central nervous system malignancies.
Citation Format: Steve T. Durant, Kurt G. Pike, Nicola Colclough, Lucy Riches, Antonio Garcia-Trinidad, Thomas Hunt, Stephanie Ling, Jonathan Stott, Ian Barrett, Li Zheng, Yingchun Wang, Kan Chen, Tianwei Zhang, Venkatesh Pilla Reddy, Andrew Sykes, Peter Johnstrom, Gemma Jones, Andrew Pierce, Jeremy Karlin, Jenna Kahn, Jasmine Allen, Kristoffer Valerie, Ruth Illingworth, Martin Pass. AZD1390, a potent and selective orally bioavailable blood-brain barrier-penetrant ATM inhibitor, radiosensitizes and improves survival of orthotopic glioma and metastatic brain tumor models [abstract]. In: Proceedings of the AACR-NCI-EORTC International Conference: Molecular Targets and Cancer Therapeutics; 2017 Oct 26-30; Philadelphia, PA. Philadelphia (PA): AACR; Mol Cancer Ther 2018;17(1 Suppl):Abstract nr A104.