EPZ004777
目录号 : GC13383
A potent inhibitor of DOT1L
Cas No.:1338466-77-5
Sample solution is provided at 25 µL, 10mM.
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Quality Control & SDS
- View current batch:
- Purity: >98.00%
- COA (Certificate Of Analysis)
- SDS (Safety Data Sheet)
- Datasheet
| Cell experiment [1]: | |
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Cell lines |
RAW264.7 cells |
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Preparation Method |
DOT1L enzyme inhibition enhances OC fusion and resorption ability a RAW264.7 cells pretreated with DMSO or the indicated concentrations of DOT1L inhibitors (EPZ5676 and EPZ004777) and stimulated with RANKL for 60 h. OCs were fixed and stained for TRAP. |
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Reaction Conditions |
1 and 10 µM; 5 days |
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Applications |
Treatment with DOT1L inhibitors (EPZ5676 and EPZ004777) increased the proportion and number of large OCs, which was approximately twice that observed in the control group. Furthermore, no significant difference was observed between the effects of treatment at 1 and 10 µM of the DOT1L inhibitors. |
| Animal experiment [2]: | |
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Animal models |
BALB/c-nu mice |
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Preparation Method |
1 × 106 HCT116 cells were subcutaneously injected in the left flank of the BALB/c-nu mice. After tumor pumped, the mice were randomly divided into two groups. One group was treated with PBS with 10% DMSO, while the other group was treated with EPZ004777 (100 mg/kg/day, diluted into PBS with 10% DMSO) for 16 days. At the termination of the experiment, tumors were removed and weighed. |
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Dosage form |
100 mg/kg/day; i.p. |
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Applications |
The results showed that the tumor volumes and weights of all EPZ004777-treated tumors in the nude mice were significantly smaller and lighter than the control groups, respectively. |
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References: [1]. Gao Y, Ge W. The histone methyltransferase DOT1L inhibits osteoclastogenesis and protects against osteoporosis. Cell Death Dis. 2018 Jan 18;9(2):33. [2]. Yang L, et al. Silencing or inhibition of H3K79 methyltransferase DOT1L induces cell cycle arrest by epigenetically modulating c-Myc expression in colorectal cancer. Clin Epigenetics. 2019 Dec 30;11(1):199. |
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EPZ004777, as a potent epigenetic modulators, can reverse TGF-β1 induced T regulatory cells and may be used to treat diverse immune disorders[1].
In vitro, EPZ004777 has concentration-dependent inhibition of DOT1L enzyme activity with an IC50 of 400 ± 100 pM. In vitro experiment it shown that in MV4-11 cells incubated with 3 μM EPZ004777, a concentration sufficient for maximal cellular DOT1L inhibition. After treatment with 1 day, there is a apparently modest reduction in H3K79me2 levels, but full depletion took 4–5 days. In vitro efficacy test it exhibited that treatment with 3 μM EPZ004777 caused a concentration-dependent decrease of both transcripts in each cell line with IC50 s of approximately 700 nM.[2] In vitro, treatment with 30 μM and 50 μM EPZ004777 obviously decreased cell viability of SW480 cells in a dose-dependent manner. Also 30 μM, 50 μM, and 70 μM EPZ004777 treatment in a dose-dependent manner inhibited the cell viability of HCT116 cells.[3] In vitro, EPZ004777 could also inhibit the proliferation and induce the differentiation of YBT-5 cells[4].
In vivo, nude mice bearing MV4-11 xenograft tumors loaded with a 50 mg/ml solution of EPZ004777, H3K79me2 levels were markedly decreased in tumors from mice treated with EPZ004777 compared to untreated controls.[2] In vivo experiment it demonstated that treatment with 10 and 50?mg/kg EPZ004777 via subconjunctival injection could alleviate corneal injury and opacity.[5].
References:
[1]Premkumar K, Shankar BS. Identification of EPZ004777 and FG2216 as inhibitors of TGF-β1 induced Treg cells by screening a library of epigenetic compounds. Life Sci. 2022 Jul 15;301:120643.
[2]Daigle SR, et al. Selective killing of mixed lineage leukemia cells by a potent small-molecule DOT1L inhibitor. Cancer Cell. 2011 Jul 12;20(1):53-65.
[3]Yang L, et al. Silencing or inhibition of H3K79 methyltransferase DOT1L induces cell cycle arrest by epigenetically modulating c-Myc expression in colorectal cancer. Clin Epigenetics. 2019 Dec 30;11(1):199.
[4]Wang Z, et al. Establishment and characterization of a DOT1L inhibitor-sensitive human acute monocytic leukemia cell line YBT-5 with a novel KMT2A-MLLT3 fusion. Hematol Oncol. 2019 Dec;37(5):617-625.
[5]Wan S, et al. Dot1l Aggravates Keratitis Induced by Herpes Simplex Virus Type 1 in Mice via p38 MAPK-Mediated Oxidative Stress. Oxid Med Cell Longev. 2021 Feb 15;2021:6612689.
EPZ004777 作为一种有效的表观遗传调节剂，可以逆转 TGF-β1 诱导的 T 调节细胞，可用于治疗多种免疫疾病[1]。
在体外，EPZ004777 对 DOT1L 酶活性具有浓度依赖性抑制作用，IC50 为 400 ± 100 pM。体外实验表明，在 MV4-11 细胞中与 3 μM EPZ004777 孵育，该浓度足以实现最大细胞 DOT1L 抑制。处理 1 天后，H3K79me2 水平明显适度降低，但完全耗尽需要 4-5 天。体外功效测试表明，用 3 μM EPZ004777 处理会导致每个细胞系中两种转录物的浓度依赖性降低，IC50 s 约为 700 nM。[2] 在体外，用 30 μM 和 50 μM EPZ004777 处理以剂量依赖性方式显着降低 SW480 细胞的细胞活力。此外，30 μM、50 μM 和 70 μM EPZ004777 剂量依赖性抑制 HCT116 细胞的细胞活力。[3] 在体外，EPZ004777 还可以抑制增殖并诱导分化YBT-5细胞[4].
在体内，荷载 MV4-11 异种移植肿瘤的裸鼠加载了 50 mg/ml EPZ004777 溶液，与未处理的对照组相比，用 EPZ004777 处理的小鼠的肿瘤中 H3K79me2 水平显着降低。[2] 体内实验表明，通过结膜下注射 10 和 50mg/kg EPZ004777 可以减轻角膜损伤和混浊。[5].
| Cas No. | 1338466-77-5 | SDF | |
| 化学名 | 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 | ||
| Canonical SMILES | CC(C)N(CCCNC(=O)NC1=CC=C(C=C1)C(C)(C)C)CC2C(C(C(O2)N3C=CC4=C3N=CN=C4N)O)O | ||
| 分子式 | C28H41N7O4 | 分子量 | 539.67 |
| 溶解度 | ≥ 27 mg/mL in DMSO, ≥ 94.6 mg/mL in EtOH with ultrasonic | 储存条件 | Store at -20°C |
| General tips | 请根据产品在不同溶剂中的溶解度选择合适的溶剂配制储备液；一旦配成溶液，请分装保存，避免反复冻融造成的产品失效。 储备液的保存方式和期限：-80°C 储存时，请在 6 个月内使用，-20°C 储存时，请在 1 个月内使用。 为了提高溶解度，请将管子加热至37℃，然后在超声波浴中震荡一段时间。 |
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| Shipping Condition | 评估样品解决方案：配备蓝冰进行发货。所有其他可用尺寸：配备RT，或根据请求配备蓝冰。 | ||
| 制备储备液 | |||
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1 mg | 5 mg | 10 mg |
| 1 mM | 1.853 mL | 9.2649 mL | 18.5298 mL |
| 5 mM | 0.3706 mL | 1.853 mL | 3.706 mL |
| 10 mM | 0.1853 mL | 0.9265 mL | 1.853 mL |
| 第一步：请输入基本实验信息（考虑到实验过程中的损耗，建议多配一只动物的药量） | ||||||||||
| 给药剂量 | mg/kg |  |
动物平均体重 | g | |
每只动物给药体积 | ul | |
动物数量 | 只 |
| 第二步：请输入动物体内配方组成（配方适用于不溶于水的药物；不同批次药物配方比例不同，请联系GLPBIO为您提供正确的澄清溶液配方） | ||||||||||
| % DMSO % % Tween 80 % saline | ||||||||||
| 计算重置 | ||||||||||
计算结果:
工作液浓度： mg/ml；
DMSO母液配制方法： mg 药物溶于 μL DMSO溶液（母液浓度 mg/mL，
体内配方配制方法：取 μL DMSO母液，加入 μL PEG300，混匀澄清后加入μL Tween 80，混匀澄清后加入 μL saline，混匀澄清。
1. 首先保证母液是澄清的；
2.
一定要按照顺序依次将溶剂加入，进行下一步操作之前必须保证上一步操作得到的是澄清的溶液，可采用涡旋、超声或水浴加热等物理方法助溶。
3. 以上所有助溶剂都可在 GlpBio 网站选购。
Identification of EPZ004777 and FG2216 as inhibitors of TGF-β1 induced Treg cells by screening a library of epigenetic compounds
Aims: Regulatory T cells play an essential role in immune tolerance and homeostasis. Their long-term stability depends on epigenetic modifications and identification of small molecule modulators of Treg differentiation therefore has many applications. In this study, we performed a novel functional screen of an epigenetic compound library to identify compounds that can modulate generation of TGF-β1 induced T regulatory cells. Main methods: A screening strategy based on IFN-γ ELISA was designed to screen epigenetic compound library. Effect of hit compounds on Treg phenotype and function was assessed by RT-PCR, flow cytometry and suppression assays. TGF-β signalling proteins were assayed by western blotting. Chromatin Immunoprecipitation (ChIP) assay was used to assess epigenetic modifications at Foxp3 gene locus. Key findings: We screened 160 compounds to identify hits capable of reversing TGF-β induced inhibition of IFN-γ production in activated spleen cells and CD4+ T cells. Two compounds EPZ004777 and FG-2216 consistently reversed TGF-β1 iTregs in terms of (a) differentiation of na?ve T cells into CD4+CD25+Foxp3+Treg cells, (b) Foxp3 target gene expression and (c) Treg suppressive function without affecting TGF-β downstream signalling. ChIP assay revealed that the compounds were able to reverse - TGF- β mediated decrease in epigenetic marks H3K27me3 and 5-mC and an increase in epigenetic marks H3K4me3 and H3K27Ac in the promoter and conserved non coding sequence (CNS1) regions of the Foxp3 gene. Significance: EPZ004777 and FG-2216 have been identified as potent epigenetic modulators that can reverse TGF-β1 induced T regulatory cells and may be used to treat diverse immune disorders.
Histone methyltransferase DOT1L coordinates AR and MYC stability in prostate cancer
The histone methyltransferase DOT1L methylates lysine 79 (K79) on histone H3 and is involved in Mixed Lineage Leukemia (MLL) fusion leukemogenesis; however, its role in prostate cancer (PCa) is undefined. Here we show that DOT1L is overexpressed in PCa and is associated with poor outcome. Genetic and chemical inhibition of DOT1L selectively impaired the viability of androgen receptor (AR)-positive PCa cells and organoids, including castration-resistant and enzalutamide-resistant cells. The sensitivity of AR-positive cells is due to a distal K79 methylation-marked enhancer in the MYC gene bound by AR and DOT1L not present in AR-negative cells. DOT1L inhibition leads to reduced MYC expression and upregulation of MYC-regulated E3 ubiquitin ligases HECTD4 and MYCBP2, which promote AR and MYC degradation. This leads to further repression of MYC in a negative feed forward manner. Thus DOT1L selectively regulates the tumorigenicity of AR-positive prostate cancer cells and is a promising therapeutic target for PCa.
Design, synthesis and in vitro anti-Zika virus evaluation of novel Sinefungin derivatives
We report herein the design and synthesis of a series of novel Sinefungin (SIN) derivatives, based on the structures of SIN and its analogue EPZ004777. Our results reveal that target compounds 1ad-af, 1ba-bb and 1bf-bh show better activity (IC50 = 4.56-20.16 μM) than EPZ004777 (IC50 = 35.19 μM). Surprisingly, SIN was founded to be not as active (IC50 > 50 μM) as we and other research groups predicted. Interestingly, the intermediates 9a-b and 11b display potent anti-ZIKV potency (IC50 = 6.33-29.98 μM), and compound 9a also exhibits acceptable cytotoxicity (CC50 > 200 μM), suggesting their promising potential to be leads for further development.
Selective killing of mixed lineage leukemia cells by a potent small-molecule DOT1L inhibitor
Mislocated enzymatic activity of DOT1L has been proposed as a driver of leukemogenesis in mixed lineage leukemia (MLL). The characterization of EPZ004777, a potent, selective inhibitor of DOT1L is reported. Treatment of MLL cells with the compound selectively inhibits H3K79 methylation and blocks expression of leukemogenic genes. Exposure of leukemic cells to EPZ004777 results in selective killing of those cells bearing the MLL gene translocation, with little effect on non-MLL-translocated cells. Finally, in vivo administration of EPZ004777 leads to extension of survival in a mouse MLL xenograft model. These results provide compelling support for DOT1L inhibition as a basis for targeted therapeutics against MLL.
Targeting human SET1/MLL family of proteins
The SET1 family of proteins, and in particular MLL1, are essential regulators of transcription and key mediators of normal development and disease. Here, we summarize the detailed characterization of the methyltransferase activity of SET1 complexes and the role of the key subunits, WDR5, RbBP5, ASH2L, and DPY30. We present new data on full kinetic characterization of human MLL1, MLL3, SET1A, and SET1B trimeric, tetrameric, and pentameric complexes to elaborate on substrate specificities and compare our findings with what has been reported before. We also review exciting recent work identifying potent inhibitors of oncogenic MLL1 function through disruption of protein-protein interactions within the MLL1 complex.