KMG-732 is a dual-target inhibitor that targets GAK and β-tubulin, with K d values of 12.0 nM and 19.3 μM, respectively. KMG-732 exhibits broad-spectrum antiproliferative activity in various human cancer cell lines and directly binds to purified tubulin in vitro. KMG-732 reduces the phosphorylation level of AP2M1, induces G 2 /M phase cell cycle arrest, disrupts the microtubule network, and inhibits cancer cell migration and invasion. KMG-732 shows significant antitumor activity in organoid and in vivo xenograft models. KMG-732 can be used for cancer research [1].
In Vivo, KMG-732 (0.5 mg/kg; i.p.; once every 2 days; for 16 consecutive days) reduces tumor volume by 67.5% in HCT116 xenograft mice without affecting body weight[1]. No significant toxicity or mortality is observed in C57BL/6 mice treated with KMG-732 (5-20 mg/kg; i.p.; single administration)[1].
In Vitro, KMG-732 (10 μM; 90 min) potently inhibits the polymerization of purified tubulin in vitro, with a Vmax of 13.95 mO.D./min[1]. KMG-732 (0.78-12.5 μM; 300 sec) exhibits binding affinity for purified tubulin in vitro, with a Kd value of 19.3 μM[1]. KMG-732 (1000 nM; 1 h) binds selectively and potently to purified GAK kinase in vitro with a Kd of 12.0 nM, while exhibiting extremely weak binding affinity toward most other tested kinases[1]. KMG-732 potently inhibits the activity of purified GAK kinase in vitro, with an IC50 of 73 nM[1]. KMG-732 (serial concentrations; 72 h) exhibits broad-spectrum antiproliferative activity against various human cancer cell lines in vitro: the GI50 values against pancreatic cancer Mia-paca2, KP2, and BxPC3 cells are 2.97, 10.84, and 16.46 nM, respectively; the GI50 values against colorectal cancer HCT116 and DLD-1 cells are 20.15 and 16.62 nM, respectively; the GI50 values against lung cancer A549 and H522 cells are 8.54 and 9.37 nM, respectively; the GI50 values against liver cancer SK-HEP-1 and HepG2 cells are 12.67 and 14.18 nM, respectively; the GI50 values against breast cancer MCF7 and MDA-MB-231 cells are 10.43 and 111.17 nM, respectively[1]. KMG-732 (0.1 μM; 1-16 h) completely disrupts the microtubule network in HCT116 colorectal cancer cells, which is characterized by perinuclear aggregation of tubulin and disappearance of cytoplasmic filamentous structures; it also induces the translocation of α-tubulin to the depolymerized (soluble) fraction, confirming its microtubule-destabilizing activity[1]. KMG-732 (1 μM; 6 h) reduces the levels of acetylated and detyrosinated α-tubulin (markers of stable microtubules) in HCT116 colorectal cancer cells within 6 hours of treatment[1]. KMG-732 (0.1-10 μM; 4 h pre-incubation plus 2 h EBI treatment) competes with the colchicine-site probe EBI for binding to β-tubulin in HCT116 colorectal cancer cells[1]. KMG-732 (10-100 nM) induces changes in the levels of mitotic regulatory proteins in HCT116 colorectal cancer cells (decreased Wee1 expression, increased Aurora A, Aurora B and Cyclin B1 expression), indicating the occurrence of mitotic arrest[1]. KMG-732 (0.1 μM; 2-8 h) induces time-dependent G2/M phase arrest in HCT116, Mia-paca2 and BxPC3 cancer cells in vitro[1]. KMG-732 (10 nM; 24 h) significantly inhibits migration and invasion of HCT116 colorectal cancer cells in vitro, reducing both migratory and invasive capacities by over 40%, and this effect occurs without a decrease in cell viability[1]. KMG-732 (0.1-1000 nM; 2 weeks) potently inhibits long-term colony formation of HCT116 colorectal cancer cells in vitro, with complete inhibition achieved at concentrations ≥100 nM[1]. KMG-732 (72 h) maintains potent antiproliferative activity in both parental K562 leukemia cells (GI50 = 20.7 nM) and P-gp-overexpressing K562A leukemia cells (GI50 = 14.8 nM). KMG-732 (0.002-10 μM) exhibits anti-tumor activity in intestinal tumor organoids derived from Apc^(1638n/+) mice, with an IC50 of 0.021 μM[1].
References:
[1]. Park S, et al. Discovery and Pharmacological Characterization of a Dual-Acting 2-Amino-3-Cyanopyridine Inhibitor Targeting Microtubule Polymerization and GAK. Journal of medicinal chemistry. 2026 Jul 09;69(13):15489-15509.
In Vivo, KMG-732 (0.5 mg/kg; i.p.; once every 2 days; for 16 consecutive days) reduces tumor volume by 67.5% in HCT116 xenograft mice without affecting body weight[1]. No significant toxicity or mortality is observed in C57BL/6 mice treated with KMG-732 (5-20 mg/kg; i.p.; single administration)[1].
In Vitro, KMG-732 (10 μM; 90 min) potently inhibits the polymerization of purified tubulin in vitro, with a Vmax of 13.95 mO.D./min[1]. KMG-732 (0.78-12.5 μM; 300 sec) exhibits binding affinity for purified tubulin in vitro, with a Kd value of 19.3 μM[1]. KMG-732 (1000 nM; 1 h) binds selectively and potently to purified GAK kinase in vitro with a Kd of 12.0 nM, while exhibiting extremely weak binding affinity toward most other tested kinases[1]. KMG-732 potently inhibits the activity of purified GAK kinase in vitro, with an IC50 of 73 nM[1]. KMG-732 (serial concentrations; 72 h) exhibits broad-spectrum antiproliferative activity against various human cancer cell lines in vitro: the GI50 values against pancreatic cancer Mia-paca2, KP2, and BxPC3 cells are 2.97, 10.84, and 16.46 nM, respectively; the GI50 values against colorectal cancer HCT116 and DLD-1 cells are 20.15 and 16.62 nM, respectively; the GI50 values against lung cancer A549 and H522 cells are 8.54 and 9.37 nM, respectively; the GI50 values against liver cancer SK-HEP-1 and HepG2 cells are 12.67 and 14.18 nM, respectively; the GI50 values against breast cancer MCF7 and MDA-MB-231 cells are 10.43 and 111.17 nM, respectively[1]. KMG-732 (0.1 μM; 1-16 h) completely disrupts the microtubule network in HCT116 colorectal cancer cells, which is characterized by perinuclear aggregation of tubulin and disappearance of cytoplasmic filamentous structures; it also induces the translocation of α-tubulin to the depolymerized (soluble) fraction, confirming its microtubule-destabilizing activity[1]. KMG-732 (1 μM; 6 h) reduces the levels of acetylated and detyrosinated α-tubulin (markers of stable microtubules) in HCT116 colorectal cancer cells within 6 hours of treatment[1]. KMG-732 (0.1-10 μM; 4 h pre-incubation plus 2 h EBI treatment) competes with the colchicine-site probe EBI for binding to β-tubulin in HCT116 colorectal cancer cells[1]. KMG-732 (10-100 nM) induces changes in the levels of mitotic regulatory proteins in HCT116 colorectal cancer cells (decreased Wee1 expression, increased Aurora A, Aurora B and Cyclin B1 expression), indicating the occurrence of mitotic arrest[1]. KMG-732 (0.1 μM; 2-8 h) induces time-dependent G2/M phase arrest in HCT116, Mia-paca2 and BxPC3 cancer cells in vitro[1]. KMG-732 (10 nM; 24 h) significantly inhibits migration and invasion of HCT116 colorectal cancer cells in vitro, reducing both migratory and invasive capacities by over 40%, and this effect occurs without a decrease in cell viability[1]. KMG-732 (0.1-1000 nM; 2 weeks) potently inhibits long-term colony formation of HCT116 colorectal cancer cells in vitro, with complete inhibition achieved at concentrations ≥100 nM[1]. KMG-732 (72 h) maintains potent antiproliferative activity in both parental K562 leukemia cells (GI50 = 20.7 nM) and P-gp-overexpressing K562A leukemia cells (GI50 = 14.8 nM). KMG-732 (0.002-10 μM) exhibits anti-tumor activity in intestinal tumor organoids derived from Apc^(1638n/+) mice, with an IC50 of 0.021 μM[1].
References:
[1]. Park S, et al. Discovery and Pharmacological Characterization of a Dual-Acting 2-Amino-3-Cyanopyridine Inhibitor Targeting Microtubule Polymerization and GAK. Journal of medicinal chemistry. 2026 Jul 09;69(13):15489-15509.