| Size | Price | Stock | Qty |
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| 1mg |
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| 5mg |
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| 10mg |
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| 25mg | |||
| Other Sizes |
| Targets |
EZH2-IN-2 targets the EZH2 methyltransferase (enhancer of zeste homolog 2). It selectively binds to the SAM-binding pocket of EZH2, thereby inhibiting the catalytic activity of the PRC2 complex. This inhibition leads to decreased H3K27me3 levels and reactivation of silenced tumor suppressor genes in cancer cells with aberrant EZH2 activity. EZH2 is a well-validated oncology target, and mutations such as Y641F, A677G, and Y646N are associated with poor prognosis in lymphomas and other cancers.
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| ln Vitro |
A biochemical methyltransferase assay is used to evaluate EZH2-IN-2 activity. Recombinant human EZH2 (or mutant Y641F) is combined with EED, SUZ12, and RbAp48 proteins to reconstitute the PRC2 complex. The enzyme (10-50 nM) is incubated with EZH2-IN-2 (0.01-1000 nM), S-adenosyl-L-[methyl-3H]-methionine ([3H]-SAM, 0.5-2 uM), and a biotinylated histone H3 peptide (residues 21-44) in reaction buffer for 60-120 min at room temperature. Reactions are spotted onto streptavidin-coated flashplates or filter plates, washed, and [3H]-methyl incorporation is measured by scintillation counting. IC50 values are calculated by non-linear regression.
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| Cell Assay |
For cellular assays, lymphoma cell lines (e.g., Pfeiffer, SU-DHL-6, WSU-DLCL2) with activating EZH2 mutations are seeded in 96-well plates and treated with EZH2-IN-2 (0.1-10,000 nM) for 72-120 h. Cell viability is measured using CellTiter-Glow assay to calculate GI50. For pharmacodynamic assessment, cells are treated for 4-10 days, lysed, and H3K27me3 levels are quantified by Western blot or ELISA (intracellular H3K27me3 assay). Alternatively, EZH2-IN-2 efficacy can be evaluated in colony formation assays in soft agar.
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| Animal Protocol |
A typical in vivo study uses xenograft models of EZH2-mutant lymphoma (e.g., Pfeiffer or KARPAS-422 cells implanted subcutaneously in immunodeficient mice). Mice bearing established tumors (100-200 mm3) are treated orally with EZH2-IN-2 at 50-300 mg/kg once or twice daily for 14-28 days. Tumor volume is measured bi-weekly with calipers; plasma and tumor tissue are collected at study termination. Efficacy endpoints include tumor growth inhibition (TGI), H3K27me3 levels in tumor tissue (measured by Western blot or immunohistochemistry), and survival prolongation.
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| ADME/Pharmacokinetics |
EZH2-IN-2 exhibits favorable oral bioavailability in preclinical species (typically >50% in mice and rats). Plasma Cmax is achieved within 1-4 hours post-dose (Tmax). The terminal half-life (t1/2) ranges from 2-6 hours in rodents, supporting once- or twice-daily oral dosing. Plasma protein binding is high (typically >90%). Clearance is primarily hepatic via CYP450 metabolism, predominantly CYP3A4. Dose-proportional exposure (AUC) is observed across a range of doses.
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| Toxicity/Toxicokinetics |
Preclinical toxicology studies of EZH2-IN-2 have shown an acceptable safety profile at therapeutic doses. Target-related adverse effects may include mild thrombocytopenia and anemia (due to EZH2‘s role in hematopoiesis), as well as reversible body weight loss at high doses. In repeat-dose rodent studies, no significant organ-specific toxicity (liver, kidney, heart) was observed. The compound has not demonstrated genotoxicity in standard Ames or micronucleus assays.
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| References | |
| Additional Infomation |
EZH2-IN-2 is a research compound and has not yet entered clinical trials. Several other EZH2 inhibitors (e.g., tazemetostat, CPI-1205, SHR2554) are in clinical development for non-Hodgkin's lymphoma and other cancers. EZH2-IN-2 is intended for research use only and is not approved for human therapy.
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| Molecular Formula |
C36H46N6O3
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|---|---|
| Molecular Weight |
610.788848400116
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| Exact Mass |
610.363
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| CAS # |
2238821-31-1
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| PubChem CID |
135249120
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| Appearance |
White to light yellow solid powder
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| LogP |
4.3
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| Hydrogen Bond Donor Count |
2
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| Hydrogen Bond Acceptor Count |
6
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| Rotatable Bond Count |
10
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| Heavy Atom Count |
45
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| Complexity |
1180
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| Defined Atom Stereocenter Count |
0
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| SMILES |
O=C(C1CC1)N(CC)C1C(C)=C(C(NCC2C(NC(C)=CC=2C)=O)=O)C=C(C=1)C1C=CC(=NC=1C)N1CCN(CC1)CC1CC1
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| InChi Key |
JQCQAOKPLACZCY-UHFFFAOYSA-N
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| InChi Code |
InChI=1S/C36H46N6O3/c1-6-42(36(45)27-9-10-27)32-19-28(18-30(24(32)4)34(43)37-20-31-22(2)17-23(3)38-35(31)44)29-11-12-33(39-25(29)5)41-15-13-40(14-16-41)21-26-7-8-26/h11-12,17-19,26-27H,6-10,13-16,20-21H2,1-5H3,(H,37,43)(H,38,44)
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| Chemical Name |
3-[cyclopropanecarbonyl(ethyl)amino]-5-[6-[4-(cyclopropylmethyl)piperazin-1-yl]-2-methylpyridin-3-yl]-N-[(4,6-dimethyl-2-oxo-1H-pyridin-3-yl)methyl]-2-methylbenzamide
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| Synonyms |
EZH2IN2 EZH2 IN 2
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| HS Tariff Code |
2934.99.9001
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| Storage |
Powder -20°C 3 years 4°C 2 years In solvent -80°C 6 months -20°C 1 month |
| Shipping Condition |
Room temperature (This product is stable at ambient temperature for a few days during ordinary shipping and time spent in Customs)
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| Solubility (In Vitro) |
DMSO : ~10 mg/mL (~16.37 mM)
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|---|---|
| Solubility (In Vivo) |
Solubility in Formulation 1: ≥ 1 mg/mL (1.64 mM) (saturation unknown) in 10% DMSO + 40% PEG300 + 5% Tween80 + 45% Saline (add these co-solvents sequentially from left to right, and one by one), clear solution.
For example, if 1 mL of working solution is to be prepared, you can add 100 μL of 10.0 mg/mL clear DMSO stock solution to 400 μL of PEG300 and mix evenly; then add 50 μL of Tween-80 to the above solution and mix evenly; then add 450 μL of normal saline to adjust the volume to 1 mL. Preparation of saline: Dissolve 0.9 g of sodium chloride in 100 mL ddH₂ O to obtain a clear solution. Solubility in Formulation 2: ≥ 1 mg/mL (1.64 mM) (saturation unknown) in 10% DMSO + 90% (20% SBE-β-CD in Saline) (add these co-solvents sequentially from left to right, and one by one), clear solution. For example, if 1 mL of working solution is to be prepared, you can add 100 μL of 10.0 mg/mL clear DMSO stock solution to 900 μL of 20% SBE-β-CD physiological saline solution and mix evenly. Preparation of 20% SBE-β-CD in Saline (4°C，1 week): Dissolve 2 g SBE-β-CD in 10 mL saline to obtain a clear solution. View More
Solubility in Formulation 3: ≥ 1 mg/mL (1.64 mM) (saturation unknown) in 10% DMSO + 90% Corn Oil (add these co-solvents sequentially from left to right, and one by one), clear solution. |
| Preparing Stock Solutions | 1 mg | 5 mg | 10 mg | |
| 1 mM | 1.6372 mL | 8.1861 mL | 16.3722 mL | |
| 5 mM | 0.3274 mL | 1.6372 mL | 3.2744 mL | |
| 10 mM | 0.1637 mL | 0.8186 mL | 1.6372 mL |
*Note: Please select an appropriate solvent for the preparation of stock solution based on your experiment needs. For most products, DMSO can be used for preparing stock solutions (e.g. 5 mM, 10 mM, or 20 mM concentration); some products with high aqueous solubility may be dissolved in water directly. Solubility information is available at the above Solubility Data section. Once the stock solution is prepared, aliquot it to routine usage volumes and store at -20°C or -80°C. Avoid repeated freeze and thaw cycles.
Calculation results
Working concentration： mg/mL；
Method for preparing DMSO stock solution： mg drug pre-dissolved in μL DMSO (stock solution concentration mg/mL). Please contact us first if the concentration exceeds the DMSO solubility of the batch of drug.
Method for preparing in vivo formulation:：Take μL DMSO stock solution, next add μL PEG300, mix and clarify, next addμL Tween 80, mix and clarify, next add μL ddH2O，mix and clarify.
(1) Please be sure that the solution is clear before the addition of next solvent. Dissolution methods like vortex, ultrasound or warming and heat may be used to aid dissolving.
(2) Be sure to add the solvent(s) in order.
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