EZM8266 is an orally active and selective G9a ( EHMT2 ) histone methyltransferase inhibitor with a human EHMT2 IC50 of 1 pM. EZM8266 reduces repressive H3K9me2 marks at immune-stimulatory gene and endogenous retroviral element promoters. EZM8266 reduces colony formation, migration, and invasion of cancer cells. EZM8266 enhances IFN-γ response, increases MHC class I expression, and enhances CXCL10 -mediated T cell recruitment in cancer cells. EZM8266 can be used for the research of hepatocellular carcinoma [1] [2].
In Vivo, EZM8266 (Compound 5R) (300 mg/kg; p.o.; daily) significantly reduces tumor growth and increases intratumoral CD8+ and CD4+ T cell infiltration in an orthotopic murine HCC model[1]. EZM8266 (300 mg/kg; p.o.; 5 days per week; 4 weeks) monotherapy significantly reduces tumor burden and incidence in an orthotopic murine HCC model, and combination with anti-PD1 further enhances antitumor efficacy, normalizes serum liver enzyme levels, and increases intratumoral CD8+ and CD4+ T cell infiltration[1]. EZM8266 (300 mg/kg; p.o.; 5 days per week; 4 weeks) monotherapy significantly reduces tumor burden in a MYC/β-catenin-driven murine HCC model, and combination with anti-PD1 enhances antitumor efficacy and reshapes the tumor microenvironment to a less immunosuppressive state[1].
In Vitro, EZM8266 (Compound 5R) (0.5-10 μM; ~15 days) potently inhibits colony formation of human PLC/PRF/5 HCC cells[1]. EZM8266 (0.5-10 μM; 3 weeks) dose-dependently inhibits anchorage-independent growth of human PLC/PRF/5 HCC cells[1]. EZM8266 (1-5 μM; 24 h) dose-dependently inhibits migration of human HuH7 HCC cells[1]. EZM8266 (1-5 μM; 24 h) dose-dependently inhibits invasion of human HuH7 HCC cells[1]. EZM8266 (compound 5a) (5 μM; 48 h) induces broad transcriptional reprogramming in human PLC/PRF/5 HCC cells, upregulating immune-related pathways including innate immune response, interferon signaling, and dsRNA processing[1]. EZM8266 (5 mM; 24 h pre-treatment, 48 h single agent treatment) enhances IFN-γ-mediated upregulation of immune-related genes (including chemokines, MHC class I components, and interferon response genes) in murine PM299L HCC cells when used alone or in combination with IFN-γ[1]. EZM8266 (1 μM); 24 h pre-treatment, 48 h single agent treatment) synergizes with IFN-γ to significantly increase CXCL10 secretion in murine PM299L and human HuH7 HCC cells when used in combination[1]. EZM8266 (5 mM; 24 h pre-treatment, 48 h single agent treatment) enhances IFN-γ-mediated upregulation of MHC class I surface expression in murine PM299L and human HuH7 HCC cells when used alone or in combination with IFN-γ[1]. EZM8266 (48 h) induces significant accumulation of intracellular double-stranded RNA in murine and human HCC cells[1].
References:
[1]. Adan-Villaescusa E, et al. Histone methyl-transferase G9a inhibition boosts the efficacy of immune checkpoint inhibitors in experimental hepatocellular carcinoma. Cell Rep Med. 2026;7(4):102717.
[2]. Campbell JE, et al. Amine-substituted heterocyclic compounds as ehmt2 inhibitors, salts thereof, and methods of synthesis thereof. United States, WO2019079540 A. 2017-10-18.
In Vivo, EZM8266 (Compound 5R) (300 mg/kg; p.o.; daily) significantly reduces tumor growth and increases intratumoral CD8+ and CD4+ T cell infiltration in an orthotopic murine HCC model[1]. EZM8266 (300 mg/kg; p.o.; 5 days per week; 4 weeks) monotherapy significantly reduces tumor burden and incidence in an orthotopic murine HCC model, and combination with anti-PD1 further enhances antitumor efficacy, normalizes serum liver enzyme levels, and increases intratumoral CD8+ and CD4+ T cell infiltration[1]. EZM8266 (300 mg/kg; p.o.; 5 days per week; 4 weeks) monotherapy significantly reduces tumor burden in a MYC/β-catenin-driven murine HCC model, and combination with anti-PD1 enhances antitumor efficacy and reshapes the tumor microenvironment to a less immunosuppressive state[1].
In Vitro, EZM8266 (Compound 5R) (0.5-10 μM; ~15 days) potently inhibits colony formation of human PLC/PRF/5 HCC cells[1]. EZM8266 (0.5-10 μM; 3 weeks) dose-dependently inhibits anchorage-independent growth of human PLC/PRF/5 HCC cells[1]. EZM8266 (1-5 μM; 24 h) dose-dependently inhibits migration of human HuH7 HCC cells[1]. EZM8266 (1-5 μM; 24 h) dose-dependently inhibits invasion of human HuH7 HCC cells[1]. EZM8266 (compound 5a) (5 μM; 48 h) induces broad transcriptional reprogramming in human PLC/PRF/5 HCC cells, upregulating immune-related pathways including innate immune response, interferon signaling, and dsRNA processing[1]. EZM8266 (5 mM; 24 h pre-treatment, 48 h single agent treatment) enhances IFN-γ-mediated upregulation of immune-related genes (including chemokines, MHC class I components, and interferon response genes) in murine PM299L HCC cells when used alone or in combination with IFN-γ[1]. EZM8266 (1 μM); 24 h pre-treatment, 48 h single agent treatment) synergizes with IFN-γ to significantly increase CXCL10 secretion in murine PM299L and human HuH7 HCC cells when used in combination[1]. EZM8266 (5 mM; 24 h pre-treatment, 48 h single agent treatment) enhances IFN-γ-mediated upregulation of MHC class I surface expression in murine PM299L and human HuH7 HCC cells when used alone or in combination with IFN-γ[1]. EZM8266 (48 h) induces significant accumulation of intracellular double-stranded RNA in murine and human HCC cells[1].
References:
[1]. Adan-Villaescusa E, et al. Histone methyl-transferase G9a inhibition boosts the efficacy of immune checkpoint inhibitors in experimental hepatocellular carcinoma. Cell Rep Med. 2026;7(4):102717.
[2]. Campbell JE, et al. Amine-substituted heterocyclic compounds as ehmt2 inhibitors, salts thereof, and methods of synthesis thereof. United States, WO2019079540 A. 2017-10-18.