TNIK-IN-3 is a highly potent and orally active inhibitor of Traf2- and Nck-interacting protein kinase (TNIK). It exhibits a strong selectivity for TNIK with an IC50 value of 0.026 μM. Additionally, TNIK-IN-3 demonstrates inhibitory activity against Flt4 (IC50 = 0.030 μM), Flt1 (IC50 = 0.191 μM), and DRAK1 (IC50 = 0.411 μM). This compound holds potential for carrying out research on colorectal cancer (CRC) [1].

TNIK-IN-3 (compound 21k) exhibits inhibitory activity against Aurora-A, GCK, and MLK3 with IC50 values of 0.517 μM, 3.657 μM, and 4.552 μM, respectively [1]. It reduces HCT116 and DLD-1 cell viability dose-dependently, with IC50s of 4.26 μM and 8.00 μM at 0.1 to 100 μM over three days [1]. The compound inhibits colony formation over 10 days (2.5 to 40 μM) and cell migration over 48 hours (5 to 20 μM) [1]. TNIK-IN-3 also dose-dependently decreases LRP5 and LRP6 protein expression, as well as the Wnt target genes AXIN2 and c-Myc in HCT116 cells (5 to 40 μM, 48 hours), and significantly reduces JNK1/2 phosphorylation in Hela cells (5 to 20 μM, 48 hours) [1].

TNIK-IN-3 (compound 21k), administered orally at dosages of 100-150 mg/kg twice daily for 18 days, dose-dependently reduces tumor growth in six-week-old female NOD-SCID mice injected with HCT116 cells[1]. The higher dose of 150 mg/kg resulted in significant tumor suppression without causing weight loss or any noticeable side effects.