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Updated: Aug 26, 2025
A Method for Screening and Validation of Resistant Mutations Against Kinase Inhibitors
Published on: December 7, 2014
Prediction of Resistance Mutations Against Upcoming Anaplastic Lymphoma Kinase Inhibitors
Yuta Doi1, Hiroaki Tagaya1, Ayaka Noge1
1Department of Life Science and Medical Bioscience, Graduate School of Advanced Science and Engineering, Waseda University, 2-2 Wakamatsu-cho, Shinjuku-ku, Tokyo, 162-8480, Japan.
Background:
Chromosomal aberrations involving the anaplastic lymphoma kinase (ALK) gene have been observed in approximately 4% of patients with non-small cell lung cancer (NSCLC). Although these patients clinically benefit from treatment with various ALK tyrosine kinase inhibitors (ALK-TKIs), none of these can inhibit the development of resistance mutations. Considering inevitable drug resistance and the variety of available ALK-TKIs, it is necessary to predict the pattern of drug-resistance mutations to determine the optimal treatment strategy.
Objective:
We aimed to establish a polymerase chain reaction (PCR)-based system to predict the development of resistance mutations against ALK-TKIs and identify therapeutic strategies using the upcoming ALK-TKIs repotrectinib (TPX-0005) and ensartinib (X-396) following recurrence on first-line alectinib treatment for ALK-positive NSCLC.
Methods:
An error-prone PCR-based method for predicting drug resistance mutations was established and the half-maximal inhibitory concentration (IC50) values of the predicted ALK mutations were evaluated in a Ba/F3 cell-based assay.
Results:
We predicted several resistance mutations against repotrectinib and ensartinib, and demonstrated that the next-generation ALK-TKI TPX-0131, was active against repotrectinib-resistant mutations and that the FLT3 inhibitor gilteritinib was active against ensartinib-resistant mutations.
Conclusions:
We developed a PCR-based system for predicting drug resistance mutations. When this system was applied to repotrectinib and ensartinib, the results suggested that these drugs can be used for the second-line treatment of ALK-positive NSCLC. Predicting resistance mutations against TKIs will provide useful information to aid in the development of effective therapeutic strategies.
Insights
A new PCR-based system predicts anaplastic lymphoma kinase (ALK) tyrosine kinase inhibitor (TKI) resistance mutations in non-small cell lung cancer (NSCLC). This aids in selecting optimal treatments like repotrectinib and ensartinib for resistant ALK-positive NSCLC patients.
Area of Science:
- Oncology
- Molecular Biology
- Pharmacology
Background:
- Anaplastic lymphoma kinase (ALK) gene rearrangements occur in ~4% of non-small cell lung cancer (NSCLC) patients.
- ALK tyrosine kinase inhibitors (TKIs) are effective but resistance mutations inevitably develop.
- Predicting resistance patterns is crucial for optimizing treatment strategies in ALK-positive NSCLC.
Purpose of the Study:
- To develop a polymerase chain reaction (PCR)-based system for predicting ALK-TKI resistance mutations.
- To evaluate therapeutic strategies using novel ALK-TKIs repotrectinib and ensartinib after alectinib resistance.
- To identify effective treatments for ALK-positive NSCLC with acquired resistance.
Main Methods:
- Established an error-prone PCR method to predict drug resistance mutations.
- Assessed half-maximal inhibitory concentration (IC50) values of predicted ALK mutations in a Ba/F3 cell assay.
- Evaluated the efficacy of repotrectinib, ensartinib, TPX-0131, and gilteritinib against resistant mutations.
Main Results:
- Several resistance mutations against repotrectinib and ensartinib were predicted.
- The next-generation ALK-TKI TPX-0131 showed activity against repotrectinib-resistant mutations.
- The FLT3 inhibitor gilteritinib demonstrated activity against ensartinib-resistant mutations.
Conclusions:
- A PCR-based system for predicting drug resistance mutations was successfully developed.
- Repotrectinib and ensartinib show potential as second-line treatments for ALK-positive NSCLC.
- Predicting TKI resistance mutations is vital for developing effective therapeutic strategies in NSCLC.
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