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Brigatinib, a newly discovered AXL inhibitor, suppresses AXL-mediated acquired resistance to osimertinib in EGFR-mutated non-small cell lung cancer

Acta Pharmacologica Sinica (2024)Cite this article

Abstract

In addition to the classical resistance mechanisms, receptor tyrosine-protein kinase AXL is a main mechanism of resistance to third-generation epidermal growth factor receptor-tyrosine kinase inhibitor (EGFR-TKI) osimertinib in EGFR-mutated non-small cell lung cancer (NSCLC). Developing an effective AXL inhibitor is important to sensitize osimertinib in clinical application. In this study we assessed the efficacy of brigatinib, a second-generation of anaplastic lymphoma kinase (ALK)-TKI, as a novel AXL inhibitor, in overcoming acquired resistance to osimertinib induced by AXL activation. We established an AXL-overexpression NSCLC cell line and conducted high-throughput screening of a small molecule chemical library containing 510 anti-tumor drugs. We found that brigatinib potently inhibited AXL expression, and that brigatinib (0.5 μM) significantly enhanced the anti-tumor efficacy of osimertinib (1 μM) in AXL-mediated osimertinib-resistant NSCLC cell lines in vitro. We demonstrated that brigatinib had a potential ability to bind AXL kinase protein and further inhibit its downstream pathways in NSCLC cell lines. Furthermore, we revealed that brigatinib might decrease AXL expression through increasing K48-linked ubiquitination of AXL and promoting AXL degradation in HCC827OR cells and PC-9OR cells. In AXL-high expression osimertinib-resistant PC-9OR and HCC827OR cells derived xenograft mouse models, administration of osimertinib (10 mg·kg−1·d−1) alone for 3 weeks had no effect, and administration of brigatinib (25 mg·kg−1·d−1) alone caused a minor inhibition on the tumor growth; whereas combination of osimertinib and brigatinib caused marked tumor shrinkages. We concluded that brigatinib may be a promising clinical strategy for enhancing osimertinib efficacy in AXL-mediated osimertinib-resistant NSCLC patients.

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Fig. 1: Acquired resistance caused by changes of AXL overexpression in EGFR-mutated NSCLC cells.
Fig. 2: Compound screening identified brigatinib as a potent inhibitor of AXL.
Fig. 3: Brigatinib and osimertinib combination synergistically decreases the survival of osimertinib-resistant NSCLC cell lines.
Fig. 4: Brigatinib can potentially bind to AXL.
Fig. 5: The combination of brigatinib and osimertinib promotes AXL degradation.
Fig. 6: The combination of osimertinib with brigatinib effectively inhibits the growth of osimertinib-resistant NSCLC xenografts.

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Acknowledgements

This work was supported by the National Natural Science Foundation of China [grant numbers 81972189 and 82172623], Chongqing Technology Innovation and Application Development Special Key Project[CSTB2023TIAD-STX0003], Clinical medical technology innovation ability training program [grant numbers 2019CXLCA003], Chongqing Municipal Bureau of Science and Technology[cstc2021jscx-xczxX0015], Clinical Medical Research Talent Training Program of Army Medical University [grant numbers 2018XLC1015], Chongqing Science and Technology Commission [grant numbers cstc2021jcyj-msxmX0014, CSTB2022NSCQ-MSX0941] and Chongqing graduate scientific research innovation project [grant numbers CYB22277].

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  1. These authors contributed equally: Rui Han, Cong-hua Lu, Chen Hu

Authors and Affiliations

  1. Department of Respiratory Disease, Daping Hospital, Army Medical University, Chongqing, 400042, China

    Rui Han, Cong-hua Lu, Chen Hu, Jun Kang, Cai-yu Lin, Di Wu, Wei-ling Jiang, Guo-qing Yin & Yong He

  2. Key Laboratory of Biorheological Science and Technology, Ministry of Education, College of Bioengineering, Chongqing University, Chongqing, 400044, China

    Yuan-yao Dou

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Contributions

RH, CHL and CH, contributed equally to this work; YH conceptualized the idea and designed the study; RH and CHL development of methodology; RH, CHL, CH, CY L, YYD, DW, WLJ, GQY, JK, data acquisition; RH, CHL and CH, analysis and interpretation of data (e.g., statistical analysis, biostatistics, computational analysis); RH, CHL and YH writing, reviewing, and revising the manuscript; RH, CHL and CH administrative, technical, or material support (i.e., reporting or organizing data, constructing databases); RH, YH study supervision. All authors have read and approved this article.

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Correspondence to Yong He.

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This study was approved by the Ethics Committees of the Daping Hospital and Army Medical University and conformed to the tenets of the Declaration of Helsinki.

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Han, R., Lu, Ch., Hu, C. et al. Brigatinib, a newly discovered AXL inhibitor, suppresses AXL-mediated acquired resistance to osimertinib in EGFR-mutated non-small cell lung cancer. Acta Pharmacol Sin (2024). https://doi.org/10.1038/s41401-024-01237-4

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