HK2-IN-4 is a selective hexokinase 2 ( HK2 ) inhibitor with an IC50 value of 0.79 μM and a K d value of 0.41 μM. HK2-IN-4 blocks the interaction between HK2 and voltage-dependent anion channel 1 ( VDAC1 ). HK2-IN-4 reduces lactate and ATP levels in cancer cells. HK2-IN-4 induces the production of apoptosis ( apoptosis ) markers in cancer cells, including increased p-AMPK/AMPK ratio and Bax levels, as well as decreased Bcl2 levels. HK2-IN-4 inhibits the proliferation of cancer cells with high HK2 expression. HK2-IN-4 can be used in research related to colorectal cancer and non-small cell lung cancer [1].
In Vitro, HK2-IN-4 (Compound 106) (5 μM; 30 min) potently and selectively inhibits the activity of recombinant HK2 enzyme, with an IC50 of 0.79 μM, while its IC50 values against HK1, HK3 and HK4 are 78.47 μM, 60.86 μM and 46.14 μM, respectively[1]. HK2-IN-4 (10-100 μM; 24-72 h) selectively inhibits the viability of colorectal cancer cells with high HK2 expression (SW480, LOVO, HCT116), with an IC50 value as low as 5.00 μM after 72 h under hypoxic conditions; compared with normoxic conditions, its efficacy is enhanced under hypoxic conditions, and its cytotoxicity depends on HK2 expression[1]. HK2-IN-4 (0-40 μM; 6 h) reduces mitochondrial HK2 levels in HCT116 and SW480 cells in a dose-dependent manner under hypoxic conditions[1]. Treatment with HK2-IN-4 (50 μM; 7 h) significantly reduces the colocalization level of HK2 and VDAC1 in SW480 cells and disrupts the HK2-VDAC1 interaction[1]. HK2-IN-4 (50-100 μM; 2 h) inhibits glycolysis in SW480 and HCT116 WT cells under hypoxic conditions, but exerts no such effect on HK2 KO cells, indicating that its glycolysis-inhibiting effect is HK2-dependent[1]. HK2-IN-4 (2.5-20 μM; 12-24 h) reduces lactate and ATP levels in hypoxic SW480 colorectal cancer cells in a dose-dependent manner, increases the p-AMPK/AMPK ratio, regulates the expression levels of Bax/Bcl2, and induces bioenergetic stress and apoptosis[1].
References:
[1]. Shan W, et al. AI-driven identification of a selective dual function inhibitor blocking HK2 activity and HK2-VDAC1 interaction displaying enhanced anticancer efficacy under hypoxia. Eur J Med Chem. 2026;309:118752.
In Vitro, HK2-IN-4 (Compound 106) (5 μM; 30 min) potently and selectively inhibits the activity of recombinant HK2 enzyme, with an IC50 of 0.79 μM, while its IC50 values against HK1, HK3 and HK4 are 78.47 μM, 60.86 μM and 46.14 μM, respectively[1]. HK2-IN-4 (10-100 μM; 24-72 h) selectively inhibits the viability of colorectal cancer cells with high HK2 expression (SW480, LOVO, HCT116), with an IC50 value as low as 5.00 μM after 72 h under hypoxic conditions; compared with normoxic conditions, its efficacy is enhanced under hypoxic conditions, and its cytotoxicity depends on HK2 expression[1]. HK2-IN-4 (0-40 μM; 6 h) reduces mitochondrial HK2 levels in HCT116 and SW480 cells in a dose-dependent manner under hypoxic conditions[1]. Treatment with HK2-IN-4 (50 μM; 7 h) significantly reduces the colocalization level of HK2 and VDAC1 in SW480 cells and disrupts the HK2-VDAC1 interaction[1]. HK2-IN-4 (50-100 μM; 2 h) inhibits glycolysis in SW480 and HCT116 WT cells under hypoxic conditions, but exerts no such effect on HK2 KO cells, indicating that its glycolysis-inhibiting effect is HK2-dependent[1]. HK2-IN-4 (2.5-20 μM; 12-24 h) reduces lactate and ATP levels in hypoxic SW480 colorectal cancer cells in a dose-dependent manner, increases the p-AMPK/AMPK ratio, regulates the expression levels of Bax/Bcl2, and induces bioenergetic stress and apoptosis[1].
References:
[1]. Shan W, et al. AI-driven identification of a selective dual function inhibitor blocking HK2 activity and HK2-VDAC1 interaction displaying enhanced anticancer efficacy under hypoxia. Eur J Med Chem. 2026;309:118752.