Fig. 5: Application of CurveCurator to three drug characterization assays reveals complete molecular MoA of Afatinib. Kinobead drug–target binding data.

a Significant drug–target interactions of the entire data set were identified by CurveCurator and visualized in a volcano plot (left panel; significance (−log10 p-value from recalibrated F-statistic) vs. effect size; gray dots). The chosen decision boundary (red dashed line) resulted in an FDR of 3.0%. Afatinib-target interactions are highlighted in blue, and significant Afatinib targets are labeled with their gene names. The pEC₅₀ distribution in the right panel indicates the potency of Afatinib-target binding, highlighting EGFR as the most potent target. b CTRP drug–cell viability data. Significant phenotypic responses of cell lines in the entire data set are gray, and Afatinib-responsive cell lines are blue (left panel). Cell lines can be grouped into HER-, p38 MAPK-, or other target-sensitive (right panel) based on the potency dimension. The purple band indicates the plasma concentrations achievable in Afatinib-based cancer therapies at the maximum tolerated dose. c Cellular decryptM data of Afatinib-treated and EGFR-driven A431 cells. Volcano plot (left panel; significance (−log10 p-value from recalibrated F-statistic) vs. effect size) of phosphorylation responses in the entire data set are marked in gray, and four example phosphorylation sites are shown in color. Potency plot (second panel from the left; pEC₅₀ vs. effect size) of significant phosphorylation responses (0.4% FDR; relevance score procedure) in the entire data set are marked in gray, and the same four example phosphorylation sites as in the left panel are highlighted in color. The dose–response curves for the same four examples (second panel from the right). The pEC₅₀ distribution of all significantly regulated phosphorylation sites (right panel) can be grouped into EGFR- and p38-MAPK-dependent signaling. The purple line indicates the potency of Afatinib in the phenotypic drug response assay. Source data for (a–c) are provided as a Source Data file.