EZM8266
Based on 1 Customer Validation
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.
For research use only. We do not sell to patients.
- Purity : 98.24%
- CAS No.: 2140164-84-5
- Formula: C19H27N5O3
- Molecular Weight:373.45
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Storage:
4°C, sealed storage, away from moisture
* In solvent : -80°C, 6 months; -20°C, 1 month (sealed storage, away from moisture)
To place orders, for customer services and technical support, please contact: MedChemExpress USA
Tel: 609-228-6898 E-mail: [email protected] [email protected]
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Biological Activity
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Chemical Information
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Solvent & Solubility
- Protocols
- Purity & Documentation
- References
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Help & FAQs
All Histone Methyltransferase Isoforms
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Biological Activity
Description
IC50 & Target
[1]|
G9a 1 pM (IC50) |
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].
MedChemExpress (MCE) has not independently confirmed the accuracy of these methods. They are for reference only. Further protocols information, click here.
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Cell Line:human HuH7 HCC cells
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Concentration:1 μM; 2.5 μM; 5 μM
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Incubation Time:24 h
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Result:Significantly reduced cell invasion in a dose-dependent manner, with ~60%, ~80%, and ~90% reductions at 1, 2.5, and 5 μM respectively, compared to control.
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Cell Line:murine PM299L HCC cells
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Concentration:5 μM
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Incubation Time:24 h (pre-treatment); 48 h (treatment)
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Result:Upregulated expression of Cxcl9, Cxcl10, B2m, Hla-a, Nlrc5, Mda5, Stat1, Rigi, Casp1, Lgals3bp, Tap1, and Irgm2 when used alone.
Produced a synergistic increase in expression of all these genes when combined with IFN-γ compared to either treatment alone.
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Cell Line:murine PM299L and human HuH7 HCC cells
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Concentration:1 μM
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Incubation Time:24 h (pre-treatment); 48 h (treatment)
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Result:Slightly increased CXCL10 secretion when used alone, while IFN-γ alone induced a moderate increase.
Produced a synergistic, significant increase in CXCL10 secretion when combined with IFN-γ compared to either treatment alone in both PM299L and HuH7 cells.
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Cell Line:murine and human HCC cells
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Concentration:5 mM
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Incubation Time:24 h (pre-treatment); 48 h (treatment)
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Result:Significantly increased intracellular dsRNA accumulation, with mean fluorescence per cell ~3-fold higher compared to DMSO-treated controls.
In Vivo
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].
MedChemExpress (MCE) has not independently confirmed the accuracy of these methods. They are for reference only.
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Animal Model:C57BL/6J (male, 6-8 weeks old, orthotopic implantation of PM299L murine HCC tumor fragments)[1]
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Dosage:300 mg/kg
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Administration:p.o.; daily
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Result:Significantly reduced tumor growth compared to vehicle controls.
Significantly lowered final tumor weights compared to vehicle controls.
Increased tumor-infiltrating CD8+ T cell counts to a mean of ~100 cells/mm2 (from vehicle mean ~30 cells/mm2).
Increased CD4+ T cell counts to a mean of ~100 cells/mm2 (from vehicle mean ~30 cells/mm2).
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Animal Model:C57BL/6J (male, 6-8 weeks old, intrahepatic injection of PM299L murine HCC cells)[1]
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Dosage:300 mg/kg; 300 mg/kg plus 100 μg/mouse anti-PD1
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Administration:p.o.; 5 days per week; 4 weeks
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Result:Reduced liver index (tumor burden) from a vehicle mean of ~18% to ~7% with monotherapy.
Reduced tumor incidence from 100% to 80% with monotherapy.
Further reduced liver index to ~3% with combination therapy.
Further reduced tumor incidence to 50% with combination therapy.
Reduced serum alanine transaminase (ALT), aspartate transaminase (AST), and lactate dehydrogenase (LDH) levels to levels comparable to age-matched normal mice.
Increased tumor-infiltrating CD8+ T cell counts to a mean of ~6000 cells/mm2 with monotherapy, and to ~8000 cells/mm2 with combination therapy (from vehicle mean ~1000 cells/mm2).
Increased CD4+ T cell counts to a mean of ~15000 cells/mm2 with combination therapy (from vehicle mean ~5000 cells/mm2).
Reduced tumor area from a vehicle mean of ~80 μm2 to ~20 μm2 with monotherapy, and to ~5 μm2 with combination therapy.
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Animal Model:C57BL/6J (male, 4 weeks old, hydrodynamic tail vein injection of MYC/CTNNB1-Δ90 transposon vectors)[1]
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Dosage:300 mg/kg; 300 mg/kg plus 100 μg/mouse anti-PD1
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Administration:p.o.; 5 days per week; 4 weeks
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Result:Elicited a significant antitumor response with monotherapy.
Resulted in a pronounced reduction in tumor burden with combination therapy.
Increased infiltration of CD8+ and CD4+ T lymphocytes with combination therapy.
Decreased CD4+:CD8+ ratio with combination therapy.
Reduced regulatory T cells with combination therapy.
Preserved effector memory subsets without overt T cell exhaustion with combination therapy.
Chemical Information
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CAS No. 2140164-84-5
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Appearance Solid
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Molecular Weight 373.45
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Formula C19H27N5O3
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Color White to off-white
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SMILES
CNC1=NC(NC2=CC(OC[C@@H](CN3CCC3)O)=C(C=C2)OC)=NC(C)=C1
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Shipping
Room temperature in continental US; may vary elsewhere.
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Storage
4°C, sealed storage, away from moisture
* In solvent : -80°C, 6 months; -20°C, 1 month (sealed storage, away from moisture)
Solvent & Solubility
In Vitro:
DMSO : 104 mg/mL (278.48 mM; Need ultrasonic; Hygroscopic DMSO has a significant impact on the solubility of product, please use newly opened DMSO)
Please refer to the solubility information to select the appropriate solvent. Once prepared, please aliquot and store the solution to prevent product inactivation from repeated freeze-thaw cycles.
Storage method and period of stock solution: -80°C, 6 months; -20°C, 1 month (sealed storage, away from moisture). When stored at -80°C, please use it within 6 months. When stored at -20°C, please use it within 1 month.
Please refer to the solubility information to select the appropriate solvent. Once prepared, please aliquot and store the solution to prevent product inactivation from repeated freeze-thaw cycles.
Storage method and period of stock solution: -80°C, 6 months; -20°C, 1 month (sealed storage, away from moisture). When stored at -80°C, please use it within 6 months. When stored at -20°C, please use it within 1 month.
Concentration (start) × Volume (start) = Concentration (final) × Volume (final)
Protocols
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RNA extraction experimental
By lysing cells, releasing RNA, and removing impurities such as proteins and DNA, high-purity RNA products are finally obtained. The commonly used traditional method is the guanidine isothiocyanate/phenol/chloroform method (Trizol), which is suitable for a variety of animal materials including animal tissues, microorganisms, cultured cells, etc., and most plant materials.
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Cell Cytotoxicity Assay
Cytotoxicity assays are usually based on the assessment of cell membrane damage, which can also be indirectly detected by measuring cell viability. Detection methods include MTT assay, CKK-8 assay, LDH assay and ATP assay, etc.
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Cell migration
Cell migration is a method that plays an important role in wound healing, cell differentiation, embryonic development, etc.
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Cell invasion
Cell invasion is the ability of cells to migrate from one area to another via the extracellular matrix. Cell invasion is the response of normal and cancer cells to chemical and mechanical stimuli. Before migrating to a new region, the extracellular matrix is degraded by proteases within the cell. Cell invasion often occurs during wound repair, vascularization and inflammation, abnormal tissue invasion, and tumor cell metastasis.
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Soft Agar Colony Formation Assay
Soft agar colony formation assay measures anchorage-independent growth, in which transformed or tumorigenic cells proliferate as colonies in a semisolid agar matrix while many non-transformed adherent cells fail to proliferate without attachment; classic studies showed that growth in semisolid medium correlates with tumorigenicity in nude mice, and later protocol papers describe the method as a stringent in vitro assay for malignant transformation. The readout is the number, size, morphology, or signal intensity of colonies formed within agar after incubation; published formats include manual colony counting after staining, 96-well or 384-well quantitative formats, DNA-binding dye detection, MTT/tetrazolium-based detection, digital image analysis, and PCR-based marker detection from soft agar cultures.
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Liver Cancer Modeling
Liver cancer can be classified into primary liver cancer and secondary liver cancer. Secondary liver cancer is the metastatic liver cancer. Primary liver cancer includes hepatocellular carcinoma (HCC), intrahepatic cholangiocarcinoma (ICC) and fibrolamellar HCC, of which HCC is the most common form, accounting for approximately 90% of primary liver cancers[1]. HCC mouse models include chemical agent-induced models, transplanted tumor models, and genetic engineered models.
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Colony Formation (Clonogenic) Assay
The clonogenic (colony formation) assay measures the ability of a single cell to retain reproductive viability and form a macroscopic colony, typically defined as a cluster derived from one progenitor cell after a defined growth period. This assay is widely used to evaluate cell survival after exposure to ionizing radiation or cytotoxic treatments and is considered a standard method in radiation biology for generating dose-response relationships of reproductive cell death. Colony formation reflects long-term proliferative capacity rather than short-term metabolic activity, and survival is quantified by comparing treated versus untreated conditions based on colony number and derived survival fractions.
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Research Protocol for Cancer Immunology
Cancer immunology studies how the immune system recognizes, suppresses, edits, or fails to eliminate malignant cells through tumor antigen release, antigen presentation, T-cell priming, immune trafficking, tumor-cell killing, and feedback inhibition in the tumor microenvironment. The cancer-immunity cycle links tumor antigenicity, dendritic-cell priming, CD8+ T-cell infiltration, cytotoxic function, and immune-checkpoint regulation to tumor rejection or immune escape. Immune-checkpoint pathways such as PD-1/PD-L1 and CTLA-4 suppress antitumor T-cell activity and can be therapeutically blocked, but many tumors remain resistant because of poor antigen presentation, weak T-cell infiltration, suppressive myeloid cells, regulatory T cells, and tumor-intrinsic immune-exclusion programs. Unresolved questions include which immune-cell states predict response, how tumor-intrinsic pathways exclude immune cells, how myeloid suppression limits checkpoint blockade, and which combination strategies
Purity & Documentation
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Data Sheet (282 KB)
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SDS (252 KB)
- English - EN (252 KB)
- Français - FR (252 KB)
- Deutsch - DE (252 KB)
- Norwegian - NO (252 KB)
- Español - ES (252 KB)
- Swedish - SV (252 KB)
- Italian - IT (252 KB)
- Korean - KR (252 KB)
- Portuguese - PT (252 KB)
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Handling Instructions (2659 KB)
References
Complete Stock Solution Preparation Table
Please refer to the solubility information to select the appropriate solvent. Once prepared, please aliquot and store the solution to prevent product inactivation from repeated freeze-thaw cycles.
Storage method and period of stock solution: -80°C, 6 months; -20°C, 1 month (sealed storage, away from moisture). When stored at -80°C, please use it within 6 months. When stored at -20°C, please use it within 1 month.
| Optional Solvent | Concentration Solvent Mass | 1 mg | 5 mg | 10 mg | 25 mg |
|---|---|---|---|---|---|
| DMSO | 1 mM | 2.6777 mL | 13.3887 mL | 26.7773 mL | 66.9434 mL |
| 5 mM | 0.5355 mL | 2.6777 mL | 5.3555 mL | 13.3887 mL | |
| 10 mM | 0.2678 mL | 1.3389 mL | 2.6777 mL | 6.6943 mL | |
| 15 mM | 0.1785 mL | 0.8926 mL | 1.7852 mL | 4.4629 mL | |
| 20 mM | 0.1339 mL | 0.6694 mL | 1.3389 mL | 3.3472 mL | |
| 25 mM | 0.1071 mL | 0.5355 mL | 1.0711 mL | 2.6777 mL | |
| 30 mM | 0.0893 mL | 0.4463 mL | 0.8926 mL | 2.2314 mL | |
| 40 mM | 0.0669 mL | 0.3347 mL | 0.6694 mL | 1.6736 mL | |
| 50 mM | 0.0536 mL | 0.2678 mL | 0.5355 mL | 1.3389 mL | |
| 60 mM | 0.0446 mL | 0.2231 mL | 0.4463 mL | 1.1157 mL | |
| 80 mM | 0.0335 mL | 0.1674 mL | 0.3347 mL | 0.8368 mL | |
| 100 mM | 0.0268 mL | 0.1339 mL | 0.2678 mL | 0.6694 mL |
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