Journal Article

Discovery of a novel ALK/ROS1/FAK inhibitor, APG-2449, in preclinical non-small cell lung cancer and ovarian cancer models

Zhang, Kaixiang, Wang, Shaomeng, Wang, Guangfeng, Min, Ping, Xiong, Dengkun, Fang, Douglas D., Tao, Ran, Wang, Qixin, Tang, Chunyang, Xu, Chunhua, Zhai, Yifan, Zhai, Guoqin, Wang, Jingwen, Chen, Jianyong, Yang, Dajun, Li, Yuanbao

2022

Overview

Background Tyrosine kinase inhibitors (TKIs) are mainstays of cancer treatment. However, their clinical benefits are often constrained by acquired resistance. To overcome such outcomes, we have rationally engineered APG-2449 as a novel multikinase inhibitor that is highly potent against oncogenic alterations of anaplastic lymphoma kinase ( ALK ), ROS proto-oncogene 1 receptor tyrosine kinase ( ROS1 ), and focal adhesion kinase ( FAK ). Here we present the preclinical evaluation of APG-2449, which exhibits antiproliferative activity in cells carrying ALK fusion or secondary mutations. Methods KINOMEscan® and LANCE TR-FRET were used to characterize targets and selectivity of APG-2449. Water-soluble tetrazolium salt (WST-8) viability assay and xenograft tumorigenicity were employed to evaluate therapeutic efficacy of monotherapy or drug combination in preclinical models of solid tumors. Western blot, pharmacokinetic, and flow cytometry analyses, as well as RNA sequencing were used to explore pharmacokinetic–pharmacodynamic correlations and the mechanism of actions driving drug combination synergy. Results In mice bearing wild-type or ALK/ROS1 -mutant non-small-cell lung cancer (NSCLC), APG-2449 demonstrates potent antitumor activity, with correlations between pharmacokinetics and pharmacodynamics in vivo. Through FAK inhibition, APG-2449 sensitizes ovarian xenograft tumors to paclitaxel by reducing CD44 + and aldehyde dehydrogenase 1-positive (ALDH1 + ) cancer stem cell populations, including ovarian tumors insensitive to carboplatin. In epidermal growth factor receptor ( EGFR) -mutated NSCLC xenograft models, APG-2449 enhances EGFR TKI-induced tumor growth inhibition, while the ternary combination of APG-2449 with EGFR (osimertinib) and mitogen-activated extracellular signal-regulated kinase (MEK; trametinib) inhibitors overcomes osimertinib resistance. Mechanistically, phosphorylation of ALK, ROS1, and FAK, as well as their downstream components, is effectively inhibited by APG-2449. Conclusions Taken together, our studies demonstrate that APG-2449 exerts potent and durable antitumor activity in human NSCLC and ovarian tumor models when administered alone or in combination with other therapies. A phase 1 clinical trial has been initiated to evaluate the safety and preliminary efficacy of APG-2449 in patients with advanced solid tumors, including ALK + NSCLC refractory to earlier-generation ALK inhibitors. Trial registration Clinicaltrial.gov registration: NCT03917043 (date of first registration, 16/04/2019) and Chinese clinical trial registration: CTR20190468 (date of first registration, 09/04/2019).

• Publisher: BioMed Central,BioMed Central Ltd,Springer Nature B.V,BMC
• Subject: Aldehyde dehydrogenase / Analysis / Anaplastic lymphoma kinase (ALK) / Antitumor activity / Biomedical and Life Sciences / Biomedicine / Cancer Research / Cancer therapies / Carboplatin / Care and treatment / CD44 antigen / Cell fusion / Diagnosis / Drug resistance / Drug therapy, Combination / Epidermal growth factor / Epidermal growth factor receptors / Extracellular signal-regulated kinase / Flow cytometry / Fluorescence resonance energy transfer / Focal adhesion kinase / Focal adhesion kinase (FAK) / Gene expression / Growth inhibition / Health Promotion and Disease Prevention / Inhibitor drugs / Kinases / Laboratory animals / Lung cancer / Lung cancer, Non-small cell / Medical prognosis / Medicine/Public Health / Molecular targeted therapy / Mutation / Non-small cell lung carcinoma / Oncology / Open source software / Ovarian cancer / Paclitaxel / Patient outcomes / Pharmacodynamics / Pharmacokinetics / Phosphorylation / Polyethylene glycol / Protein-tyrosine kinase receptors / Proteins / Response rates / Risk factors / RNA sequencing / ROS proto-oncogene 1 receptor tyrosine kinase (ROS1) / Small cell lung carcinoma / Solid tumors / Stem cells / Surgical Oncology / Targeted therapies / Tetrazolium salt / Tumorigenicity / Tumors / Xenografts