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Updated: Jul 4, 2026
Kinase Inhibitor Screening In Self-assembled Human Protein Microarrays
Published on: October 23, 2019
Biochemical assay-based selectivity profiling of clinically relevant kinase inhibitors on mutant forms of EGF
Edit Varkondi1, Ferenc Pinter, Kiss Robert
1Rational Drug Design Laboratory CRC, Semmelweis University, Budapest, Hungary.
Abstract:
Gefitinib and erlotinib are potent EGFR tyrosine kinase inhibitors (potentially) useful for the treatment of non-small-cell lung cancer (NSCLC). Clinical responses, however, in NSCLC patients have been linked to the presence of certain activating mutations of EGFR. We used an ELISA-based biochemical assay to confirm the selective inhibitory efficacy of gefitinib and erlotinib on the activated mutant receptor. Our results are in line with the clinical observations providing evidence for the predictive power of the kinase assay. Four additional compounds were also investigated: CI-1033 and EKB-569 had dramatic inhibitory effects on all EGFR forms, whereas PD153035 and AG1478 were active on wild-type and activating mutant protein. In docking simulations with wild-type EGFR, our inhibitory data are in good agreement with the binding scores. These data confirm that anilinoquinazolines are good starting structures for the next generation of selective drugs against mutant EGFR, whereas CI-1033 and EKB-569 may represent advances for patients with both wild-type and anilinoquinazoline-resistant mutant tumors.
Insights
Gefitinib and erlotinib selectively inhibit mutant EGFR, validating kinase assays for predicting non-small-cell lung cancer treatment response. New compounds show promise for broader EGFR inhibition.
Area of Science:
- Biochemistry
- Pharmacology
- Oncology
Background:
- Gefitinib and erlotinib are EGFR tyrosine kinase inhibitors for non-small-cell lung cancer (NSCLC).
- Clinical efficacy of these drugs correlates with specific EGFR mutations in NSCLC patients.
Purpose of the Study:
- To confirm selective inhibition of activated mutant EGFR by gefitinib and erlotinib using biochemical assays.
- To evaluate the predictive power of kinase assays for drug response.
- To investigate additional EGFR inhibitors.
Main Methods:
- ELISA-based biochemical assay to assess EGFR inhibition.
- Docking simulations to analyze drug-receptor binding.
- Evaluation of four additional compounds: CI-1033, EKB-569, PD153035, and AG1478.
Main Results:
- Gefitinib and erlotinib demonstrated selective inhibition of mutant EGFR, aligning with clinical observations.
- The kinase assay proved effective in predicting drug response.
- CI-1033 and EKB-569 inhibited all EGFR forms, while PD153035 and AG1478 targeted wild-type and mutant EGFR.
- Docking simulations supported the inhibitory data for wild-type EGFR.
Conclusions:
- Anilinoquinazolines serve as effective scaffolds for developing selective mutant EGFR inhibitors.
- CI-1033 and EKB-569 offer potential therapeutic advances for patients with wild-type or resistant EGFR mutations.
