Clk1-in-3 is a + selective and highly potent sexual Clk1 inhibitor with an IC50 of 5 nM and 300 pairs higher affinity than Dyrk1A. CLK1-IN-3 also showed highly effective inhibition of Clk2 and Clk4, with IC50 values of 42 and 108 nM, respectively. CLK1-IN-3 is effective in inducing autophagy in vitro and can be used for prevention and human treatment of acute liver injury (ALI).

CLK2:42 nM, DYRK1A:1521 nM, CLK1:5 nM, CLK4:108 nM

CLK1-IN-3 (compound 10ad) demonstrates anti-tumor potential through dual inhibition of Clk1 and Clk2. At concentrations ranging from 10 μM to 1000 μM, it effectively binds to Clk1 protein and inhibits its degradation dose-dependently. Additionally, CLK1-IN-3 (0-10 μM, 24 h) induces autophagy in Hela, BNLCL.2, and HCT 116 cells and stimulates the degradation of SQSTM1/p62 (autophagy marker)[1].

CLK1-IN-3 (0-40 mg/kg, intraperitoneal injection, single dose) significantly inhibits acute liver injury induced by acetaminophen (APAP) in the ALI model, and there is no apparent hepatocyte death observed[1].
CLK1-IN-3 (10 mg/kg; intravenous injection, oral administration, intraperitoneal injection, single dose) exhibits acceptable pharmacokinetic characteristics, with a relatively long half-life (T1/2) of 5.29 hours and an oral bioavailability of 19.5%[1].