EPZ004777

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].