| Size | Price | Stock | Qty |
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| 10mg |
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| 25mg |
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| 50mg |
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| Other Sizes |
| Targets |
EPZ004777 hydrochloride targets DOT1L (disruptor of telomeric silencing 1-like), a histone methyltransferase that catalyzes the mono-, di-, and tri-methylation of histone H3 at lysine 79 (H3K79). DOT1L is the only known enzyme responsible for H3K79 methylation, a histone mark associated with actively transcribed genes. In MLL-rearranged leukemias, DOT1L is aberrantly recruited to MLL fusion target genes, leading to sustained H3K79 methylation and constitutive expression of oncogenes such as HOXA9 and MEIS1. By inhibiting DOT1L with an IC50 of 0.4 nM, EPZ004777 hydrochloride reduces H3K79 methylation levels and down-regulates MLL fusion target gene expression, thereby selectively killing MLL-rearranged leukemia cells.
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| ln Vitro |
EPZ004777 hydrochloride, with an IC50 of 400±100 pM, has a strong concentration-suspension inhibitory action on DOT1L enzyme activity. On DOT1L, solutions of hydrochloride EPZ004777 suspend more HMTs (PRMT5, 521±137 nM; other, >50 μM). Prolonged HCl treatment with EPZ004777 has a substantial potential impact on MLL-rearranged cell lines. EPZ004777 hydrochloride markedly decreased the amount of viable MV4-11 and MOLM-13 cells, however it took some time for Jurkat cells to start growing. While Part One MV4-11 cells continued to divide when exposed to EPZ004777 hydrochloride, their population size did not alter over time, according to long-term growth curve tracking. At doses of 3 μM or above, EPZ004777 hydrochloride significantly inhibits MLL-AF9. Results MLL-AF10 and CALM-AF10 transformed cells' bone marrow stroma are inhibited by the EPZ004777 hydrochloride cycle [2].
EPZ004777 hydrochloride demonstrates potent in vitro activity as a DOT1L inhibitor. The compound has an IC50 of 0.4 nM against DOT1L, indicating exceptional potency. In cellular assays, EPZ004777 hydrochloride reduces the levels of H3K79me2 and H3K79me1 in a concentration-dependent manner. The compound selectively inhibits the proliferation of MLL-rearranged cell lines and MLL-AF9-transformed murine hematopoietic cells. EPZ004777 hydrochloride also induces differentiation and apoptosis in MLL-rearranged cells. The compound demonstrates concentration-dependent inhibition of DOT1L enzyme activity with an IC50 of 400 +/- 100 pM. It has been used to selectively kill MLL-rearranged leukemia cells in various in vitro models. |
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| ln Vivo |
The hydrochloride of EPZ004777 was well tolerated, and no appreciable toxicity was seen. Following 14 days of nonstop exposure to EPZ004777 hydrochloride, a complete blood count study revealed a substantial rise in total leukocytes as a result of an increase in neutrophils, monocytes, and lymphocytes. The hydrochloride dosage of EPZ004777 (50, 100, or 150 mg/mL) was well tolerated, and no appreciable weight loss was seen [1].
In vivo, EPZ004777 hydrochloride has demonstrated efficacy in preclinical models of MLL-rearranged leukemia. Following DOT1L inhibition, the compound selectively inhibits proliferation of MLL-AF10 and CALM-AF10-transformed murine bone marrow cells. DOT1L inhibition by EPZ004777 results in significantly decreased proliferation, decreased expression of MLL-AF6 target genes, and cell cycle arrest of MLL-AF6-transformed cells. The compound's ability to selectively target MLL-rearranged cells in vivo supports its potential for leukemia research. Comprehensive in vivo efficacy studies have been reported in the literature, demonstrating the compound's therapeutic potential in mouse models of MLL-rearranged leukemia. |
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| Enzyme Assay |
In vitro enzyme assays for EPZ004777 hydrochloride involve measuring the inhibition of DOT1L methyltransferase activity. Recombinant DOT1L enzyme is incubated with varying concentrations of the test compound, the cofactor S-adenosylmethionine (SAM), and a histone H3 peptide substrate or nucleosomes. The transfer of methyl groups to H3K79 is quantified using radiometric detection with [3H]-SAM, or by using a luminescent method such as the AlphaScreen or TR-FRET assay that detects methylated H3K79. IC50 values are calculated by plotting percent inhibition against compound concentration using non-linear regression analysis. The compound exhibits an IC50 of 0.4 nM. Selectivity assays compare the compound's activity against other histone methyltransferases. Each concentration is typically tested in duplicate or triplicate with appropriate positive and negative controls.
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| Cell Assay |
In vitro cellular assays for EPZ004777 hydrochloride are performed using MLL-rearranged leukemia cell lines (such as MV4-11, THP-1, or MOLM-13) and non-MLL-rearranged control cell lines. Cells are treated with varying concentrations of the compound for 3-7 days. Cell proliferation and viability are measured using CellTiter-Glo or MTT assays. H3K79 methylation levels (H3K79me1, H3K79me2, H3K79me3) are assessed by Western blot using methylation-specific antibodies. Expression of MLL fusion target genes (HOXA9, MEIS1) is measured by qRT-PCR. Apoptosis is assessed by annexin V/propidium iodide staining or caspase activity assays. Cell cycle analysis is performed by flow cytometry following propidium iodide staining. IC50 values for inhibition of proliferation are calculated from dose-response curves. The compound's selectivity for MLL-rearranged cells is confirmed by comparing effects on sensitive and resistant cell lines.
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| Animal Protocol |
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| ADME/Pharmacokinetics |
Pharmacokinetic properties of EPZ004777 hydrochloride have been characterized in preclinical studies. The compound has a molecular formula of C28H42ClN7O4 and a molecular weight of 576.13 g/mol. Its chemical name is 1-(3-((((2R,3S,4R,5R)-5-(4-amino-7H-pyrrolo[2,3-d]pyrimidin-7-yl)-3,4-dihydroxytetrahydrofuran-2-yl)methyl)(isopropyl)amino)propyl)-3-(4-(tert-butyl)phenyl)urea hydrochloride. The compound is soluble in DMSO at 10 mM. Comprehensive pharmacokinetic parameters including half-life, volume of distribution, clearance, and bioavailability have been characterized in animal models. The compound's pharmacokinetic profile supports its use in preclinical studies of MLL-rearranged leukemia.
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| Toxicity/Toxicokinetics |
EPZ004777 hydrochloride is intended for laboratory research use only and has not undergone comprehensive clinical toxicology testing. As a potent DOT1L inhibitor that modulates epigenetic regulation, the compound would be expected to have effects on gene expression and cell differentiation. Standard in vitro cytotoxicity assays in cell lines are typically performed alongside efficacy studies to rule out nonspecific toxicity. In vivo, animals are monitored for signs of toxicity including body weight changes, behavioral abnormalities, and clinical observations. Comprehensive toxicological characterization including genotoxicity, cardiotoxicity, and repeated-dose toxicity studies has been conducted as part of preclinical development. The compound is not approved for human use and is strictly intended for research purposes.
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| References |
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| Additional Infomation |
EPZ004777 hydrochloride is a potent and selective inhibitor of DOT1L with an IC50 of 0.4 nM. It reduces H3K79me2 and H3K79me1 levels and down-regulates HOXA9 and MEIS1 expression. The compound selectively inhibits the proliferation of MLL-rearranged leukemia cells and induces differentiation and apoptosis. EPZ004777 hydrochloride has a molecular formula of C28H42ClN7O4 and a molecular weight of 576.13 g/mol. The compound is used for leukemia research. EPZ004777 hydrochloride has not entered clinical trials and has not received regulatory approval for any indication. It is available from research chemical suppliers for non-clinical research purposes only.
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| Molecular Formula |
C₂₈H₄₂CLN₇O₄
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| Molecular Weight |
576.13
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| Exact Mass |
575.299
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| CAS # |
1380316-03-9
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| Related CAS # |
EPZ004777;1338466-77-5
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| PubChem CID |
67811385
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| Appearance |
Light yellow to yellow solid powder
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| LogP |
4.513
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| Hydrogen Bond Donor Count |
6
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| Hydrogen Bond Acceptor Count |
8
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| Rotatable Bond Count |
10
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| Heavy Atom Count |
40
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| Complexity |
788
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| Defined Atom Stereocenter Count |
4
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| SMILES |
CC(C)N(CCCNC(=O)NC1=CC=C(C=C1)C(C)(C)C)C[C@@H]2[C@H]([C@H]([C@@H](O2)N3C=CC4=C(N=CN=C43)N)O)O.Cl
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| InChi Key |
NPEPLDAFAVXSBD-XRJUUMFPSA-N
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| InChi Code |
InChI=1S/C28H41N7O4.ClH/c1-17(2)34(13-6-12-30-27(38)33-19-9-7-18(8-10-19)28(3,4)5)15-21-22(36)23(37)26(39-21)35-14-11-20-24(29)31-16-32-25(20)35;/h7-11,14,16-17,21-23,26,36-37H,6,12-13,15H2,1-5H3,(H2,29,31,32)(H2,30,33,38);1H/t21-,22-,23-,26-;/m1./s1
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| Chemical Name |
1-[3-[[(2R,3S,4R,5R)-5-(4-aminopyrrolo[2,3-d]pyrimidin-7-yl)-3,4-dihydroxyoxolan-2-yl]methyl-propan-2-ylamino]propyl]-3-(4-tert-butylphenyl)urea;hydrochloride
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| Synonyms |
EPZ 004777 HCl EPZ-004777 EPZ004777
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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 Note: Please store this product in a sealed and protected environment (e.g. under nitrogen), avoid exposure to moisture and light. |
| 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) |
H2O : ~100 mg/mL (~173.57 mM)
DMSO : ~100 mg/mL (~173.57 mM) |
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| Solubility (In Vivo) |
Solubility in Formulation 1: 100 mg/mL (173.57 mM) in PBS (add these co-solvents sequentially from left to right, and one by one), clear solution; with sonication.
(Please use freshly prepared in vivo formulations for optimal results.) |
| Preparing Stock Solutions | 1 mg | 5 mg | 10 mg | |
| 1 mM | 1.7357 mL | 8.6786 mL | 17.3572 mL | |
| 5 mM | 0.3471 mL | 1.7357 mL | 3.4714 mL | |
| 10 mM | 0.1736 mL | 0.8679 mL | 1.7357 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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