Resistance Profile and Structural Modeling of Next-Generation ROS1 Tyrosine Kinase Inhibitors

• Authors: Clare Keddy, Oregon Health and Science University; Pushkar Shinde, Emory University; Kristen Jones, Oregon Health & Science University; Stefanie Kaech, Oregon Health & Science University; Romel Somwar, Memorial Sloan Kettering Cancer Center; Ujwal Shinde, Oregon Health & Science University; Monika A Davare, Oregon Health & Science University
• Journal Title: MOLECULAR CANCER THERAPEUTICS
• Volume: Volume 21, Number 2
• Publisher: AMER ASSOC CANCER RESEARCH | 2022-02-01, Pages 336-346
• Type of Work: Article | Post-print: After Peer Review
• Publisher DOI: http://doi.org/10.1158/1535-7163.MCT-21-0395
• Subjects: Health Sciences, Oncology; Chemistry, Biochemistry
• Keywords: Science & Technology; Life Sciences & Biomedicine; Oncology; CELL LUNG-CANCER; OVERCOMES CRIZOTINIB RESISTANCE; ALK INHIBITOR; ANTITUMOR-ACTIVITY; SINGLE-ARM; OPEN-LABEL; PAN-TRK; ENTRECTINIB; MUTATION; MULTICENTER

Abstract

ROS1 fusion proteins resulting from chromosomal rearrangements of the ROS1 gene are targetable oncogenic drivers in diverse cancers. Acquired resistance to targeted inhibitors curtails clinical benefit and response durability. Entrectinib, a NTRK/ROS1/ALK targeted tyrosine kinase inhibitor (TKI), was approved for the treatment of ROS1 fusion-positive non–small cell lung cancer (NSCLC) in 2019. In addition, lorlatinib and repotrectinib are actively being explored in the setting of treatment-naïve or crizotinib-resistant ROS1 fusion driven NSCLC. Here, we employed an unbiased forward mutagenesis screen in Ba/F3 CD74-ROS1 and EZR-ROS1 cells to identify resistance liabilities to entrectinib, lorlatinib, and repotrectinib. ROS1F2004C emerged as a recurrent entrectinib resistant mutation and ROS1G2032R was discovered in entrectinib and lorlatinib-resistant clones. Cell-based and modeling data show that entrectinib is a dual type I/II mode inhibitor, and thus liable to both types of resistant mutations. Comprehensive profiling of all clinically relevant kinase domain mutations showed that ROS1L2086F is broadly resistant to all type I inhibitors, but remains sensitive to type II inhibitors. ROS1F2004C/I/V are resistant to type I inhibitors, entrectinib and crizotinib, and type II inhibitor, cabozantinib, but retain sensitivity to the type I macrocyclic inhibitors. Development of new, more selective type II ROS1 inhibitor (s) or potentially cycling type I and type II inhibitors may be one way to expand durability of ROS1-targeted agents.