Fig. 8: Efficacy and characterization of triple combination therapy in ALK1510-c4 cells and animal models.

a Relative cell viability of ALK1510-c4 cells cultured with alectinib, alectinib with dacomitinib (100 nM), alectinib with AZ6102 (100 nM), and a triple combination of alectinib + dacomitinib + AZ6102 (mean [SD] of n = 3 experiments, *P < 0.05 between doublet treatment and alectinib + dacomitinib + AZ6102 treatment; Dunnett’s test). b Caspase-3/7 activity relative to vehicle treatment was assessed in ALK1510-c4 cells treatment with the drugs described in Fig. 8a (mean [SD] of n = 3 experiments). c Immunoblots of cell lysates from ALK1510-c4 cells described in Fig. 8a after treatment for 24 h. d Mean (SD) change in tumor volume of mice bearing xenograft tumors with ALK1510-c4 cells and treated with vehicle, alectinib (2 mg/kg) + dacomitinib (10 mg/kg), alectinib (2 mg/kg) + G007-LK (10 mg/kg), and their triple combination for 14 days (P < 0.05 versus alectinib + dacomitinib (a) or versus alectinib + G007-LK (b) ; Wilcoxon rank sum test by the Holm–Bonferroni method; n = 8 mice per group). e Immunoblots of tumor lysates evaluating ALK-signaling and Wnt/β-catenin-signaling from xenograft tumors with ALK1510-c4 cells in mice treated with alectinib, alectinib + dacomitinib, alectinib + G007-LK, and their triple combination for 2 days. f Mean (SD) change from baseline in body weight of mice described in Fig. 8d. g ALK1510-c4 cells were treated with vehicle, alectinib (1000 nM), alectinib in combination with dacomitinib (100 nM) or AZ6102 (100 nM); and a triple combination of alectinib + dacomitinib + AZ6102 for 5 weeks, and then washed after 5 weeks. Cell numbers were measured using a cell counter at weeks 1, 2, 3, 4, 5, and 11. If the cell number was 1 × 10⁴ or less, it was indicated and considered a limitation of the cell counter. The cell counts were terminated at the point (†) when ALK1510-c4 cells detached from the culture flask due to over-confluence (n = 1). h ALK1510-c4 cells were sequentially treated with alectinib (1000 nM) in combination with dacomitinib (100 nM) or AZ6102 (100 nM), and a triple combination of alectinib + dacomitinib + AZ6102 for 5 weeks (left). Cell numbers were measured using a cell counter at weeks 1, 2, 3, 4, and 5 (right). If the cell number was 1 × 10⁴ or less, it was indicated and considered a limitation of the cell counter (n = 1).