c-Met-IN-2 

NCI-H1993 cell line and SNU-5 cell line are maintained in RPMI 1640 media and supplemented with 10% fetal bovine serum. NCI-H1993 cells are seeded at 5000 cells/well in 96-well plates and incubated overnight. On the next day, the cells are exposed to various concentrations of c-Met-IN-2 and further cultured for 72 h. After chromogenic reaction with CCK-8, the OD450 (with reference of OD650) is measured using a Flexstation 3 reader. IC₅₀ values are calculated using the GraphPad Prism Software. Each experiment is carried out thrice, each time in duplicate. The SNU-5 cell line assay is operated in a similar procedure as NCI-H1993 assay^[1].

MCE has not independently confirmed the accuracy of these methods. They are for reference only.

Mice^[1]
The SNU-5 at a density of 6 × 10⁶ tumor cells in 200 μL or NCI-H1993 at a density of 7 × 10⁶ tumor cells in 140 μL are injected s. c. into the right flank of nude mice. Tumor-bearing animals are sorted into groups with similar mean tumor volumes prior to treatment (usually 100-200 mm³ for SNU-5 and 150-250 mm³ for NCI-H1993). The mice are randomly assigned into control and treatment groups (n = 7 (NCI-H1993 model) or n = 6 (SNU-5 model) per group). Control groups are given vehicle alone, and treatment groups receive c-Met-IN-2 as indicated doses via oral administration once daily for 2 weeks in SNU-5 model and oral administration once daily for 3 weeks in NCI-H1993 model, respectively. The sizes of the tumors are measured twice per week using a caliper, and the tumor volume is calculated in cubic millimeter using the formula: V = (A × B²)/2, where A and B is the long and short diameters of the tumor, respectively. Body weights are monitored throughout the study as a gross measure of toxicity/morbidity. Tumor growth inhibition (TGI), expressed in percent (%), is calculated using the formula: 100% × (1-((treatedfinal day-treatedday 0)/(controlfinal day-controlday 0))). Percent tumor regression (PTR), expressed in percent (%), is calculated using the formula: 100% × (treatedday 0-treatedfinal day)/treatedday 0^[1].

MCE has not independently confirmed the accuracy of these methods. They are for reference only.

• [1]. Zhao F, et al. Identification of 3-substituted-6-(1-(1H-[1,2,3]triazolo[4,5-b]pyrazin-1-yl)ethyl)quinoline derivatives as highly potent and selective mesenchymal-epithelial transition factor (c-Met) inhibitors via metabolite profiling-based structural opt  [Content Brief]