neratinib  (MedChemExpress)

Journal: Nature Communications

Article Title: Determinants of sensitivity to HER2-targeted antibody drug conjugates in urothelial cancer

doi: 10.1038/s41467-025-67643-2

Figure Lengend Snippet: A Urothelial cancer organoids (SMBO-170, SCBO-8, SMBO-109, SMBO-114, and SMBO-106) and BT-474 ( ERBB2 amplified breast cancer) cells were treated with neratinib at the concentrations indicated for 1 h. Changes in the expression of pERK (Thr202/Tyr204), ERK, pAKT (ser473) and AKT as a function of neratinib concentration were determined by immunoblot. B Cell death quantitated by flow cytometry following 48 h of treatment with neratinib at a concentration of 100 nM. BT474: p < 0.001; SMBO-170: p = 0.0166; SCBO-8: p = 0.0177; SMBO-109: p = 0.6674; SMBO-114: p = 0.0626; SMBO-106: p = 0.3698. C Mice with established SMBO-170 or SMBO-106 xenografts were treated with neratinib (20 mg/kg; p.o. QD × 5) or T-DXd (10 mg/kg, i.v., once every 3 weeks for 9 weeks). TCN, total copy number. SMBO-170: p < 0.001 vehicle vs neratinib; p < 0.001 vehicle vs T-DXd; SMBO-106: p < 0.001 vehicle vs neratinib; p < 0.001 vehicle vs T-DXd. Two-way ANOVA test (Prism) was used for statistical analysis without adjustment. Data are presented as mean values ± SD. Only the upper error bars are displayed for clarity. D Immunoblots comparing expression of HER2 in protein lysates extracted from BT474 breast cancer cells and SMBO-170 and SMBO-106 urothelial cancer organoids and xenografts. E Cell viability as determined by MTT assays for urothelial organoids treated with T-DXd for 8 days with concentrations ranging from 0 - 16 μg/ml. F Cell viability as determined by MTT assay for organoids treated with the DNA topoisomerase 1 inhibitor exatecan (DX8951f), an analog of the T-DXd payload deruxtecan, for 3 days with concentrations ranging from 0 - 1250 nM. G Cell death quantitated by flow cytometry after 48 h of exatecan treatment (100 nM). BT474: p = 0.005; SMBO-170: p < 0.001; SMBO-106: p < 0.001. Data are representative of three independent experiments for ( A and D ). Results are shown as means ± SD for three experimental replicates for ( B and G ). P value were determined by two-sided unpaired t test with Welch’s correction (Prism) for ( B and G ). An example of the gating strategy for B and G is provided in Figure in Supplementary Information file. Significance denoted as * p < 0.05, ** p < 0.01, *** p < 0.001. ns, not significant. Source data for ( A – G ) are provided in Source Data.

Article Snippet: After 24 h to allow adherence, cells were treated with 0 or 100 nM neratinib (MedChem Express), or 0 or 100 nM exatecan (MedChem Express) with three replicates for 2 days.

Techniques: Amplification, Expressing, Concentration Assay, Western Blot, Flow Cytometry, MTT Assay