EX 527, as a SIRT1-selective inhibitor, inhibits Sirt1 ～100-fold more potently than Sirt2 and Sirt3 and has no effect on Sirt5′s deacetylation activit.The IC50 values for Sirt1 and Sirt3 are 0.09 ± 0.03 μM and 22.4 ± 2.7 μM, respectively. Sir2Tm was also efficiently inhibited by Ex-527 with IC50 of 0.9 ± 0.3 μM.[1]
In vitro experiment it shown that cell viability significantly increased and cell death decreased in cancer cells with SIRT1 silencing or?EX 527 (10 μM) treatment compared with the control after exposure to RSL3 or sulfasalazine.[2] In vitro, treatment with 1?μM EX-527 decreased colony formation of ovarian carcinoma cells, with or without overexpression of SIRT172. However, at 600?nM, EX-527 suppressed cell migration and inhibited the occurrence of epithelial–mesenchymal transition (EMT) in chemotherapy resistant oesophageal cancer cells.[4] In vitro experiment it indicated that SIRT 1 inhibition by?EX 527 (10 μM) elevated ROS production. SIRT1 and PGC-1α levels were dramatically decreased.[6]
In vivo efficacy test indicated that pharmacological blockade of Sirt-1 with 2 mg/kg EX 527/mouse/day daily for 3 weeks alleviated GVHD without impairing T-cell-mediated GVL activity.[3] In vivo efficacy study demonstrated that treatment with 5?mg/kg?EX 527 intraperitoneally in C57BL/6J mice abolished the protective effects of melatonin.[4] In vivo, the data indicated that LPS-induced intrapulmonary inflammation and LPS-induced elevation of 4E-BP1 phosphorylation were attenuated by?EX 527 (10?mg/kg, i.p.).[5]
参考文献:
[1]. Gertz M, et al. Ex-527 inhibits Sirtuins by exploiting their unique NAD+-dependent deacetylation mechanism. Proc Natl Acad Sci U S A. 2013 Jul 23;110(30):E2772-81.
[2]. Lee J, et al. Epigenetic reprogramming of epithelial-mesenchymal transition promotes ferroptosis of head and neck cancer. Redox Biol. 2020 Oct;37:101697.
[3]. Daenthanasanmak A, et al. Targeting Sirt-1 controls GVHD by inhibiting T-cell allo-response and promoting Treg stability in mice. Blood. 2019 Jan 17;133(3):266-279.
[4]. Broussy S, et al. Biochemical mechanism and biological effects of the inhibition of silent information regulator 1 (SIRT1) by EX-527 (SEN0014196 or selisistat). J Enzyme Inhib Med Chem. 2020 Dec;35(1):1124-1136.
[5]. Huang J, et al. The SIRT1 inhibitor EX-527 suppresses mTOR activation and alleviates acute lung injury in mice with endotoxiemia. Innate Immun. 2017 Nov;23(8):678-686.
[6]. Waldman M, et al. Regulation of diabetic cardiomyopathy by caloric restriction is mediated by intracellular signaling pathways involving 'SIRT1 and PGC-1α'. Cardiovasc Diabetol. 2018 Aug 2;17(1):111.
EX 527 作为 SIRT1 选择性抑制剂，抑制 Sirt1 的效力比 Sirt2 和 Sirt3 高 100 倍，并且对 Sirt5 的去乙酰化活性没有影响。Sirt1 和 Sirt3 的 IC50 值为 0.09 ± 0.03 μM 和 22.4分别为± 2.7 μM。 Sir2Tm 也被 Ex-527 有效抑制，IC50 为 0.9 ± 0.3 μM。[1]
体外实验表明，与暴露于 RSL3 或柳氮磺胺吡啶后的对照相比，SIRT1 沉默或 EX 527 (10 μM) 处理的癌细胞的细胞活力显着增加，细胞死亡减少。[2] 在体外，用 1μM EX-527 处理可减少卵巢癌细胞的集落形成，有或没有 SIRT172 过表达。然而，在 600nM 时，EX-527 抑制细胞迁移，抑制化疗耐药食管癌细胞上皮-间质转化 (EMT) 的发生。[4] 体外实验表明 SIRT 1 抑制EX 527 (10 μM) 提高了 ROS 的产量。 SIRT1 和 PGC-1α 水平显着降低。[6]
体内药效试验表明，每天 2 mg/kg EX 527/小鼠/天对 Sirt-1 进行药理学阻断，持续 3 周可减轻 GVHD，而不会损害 T 细胞介导的 GVL 活性。[3] 体内功效研究表明，在 C57BL/6J 小鼠中腹膜内注射 5 mg/kg EX 527 可消除褪黑激素的保护作用。[4] 在体内，数据表明 LPS 诱导的肺内EX 527（10mg/kg，i.p.）减弱了炎症和 LPS 诱导的 4E-BP1 磷酸化升高。[5]
| Cell experiment [1]: | |
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Cell lines |
T cells |
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Preparation Method |
Purified WT or Sirt-1?/? T cells were labeled with CFSE and cocultured with T-cell-depleted splenocytes as APCs from BALB/c mice for 5 days in the presence of either dimethyl sulfoxide or EX 527 (10 μg/mL). Cells were restimulated with phorbol 12-myristate 13-acetate and ionomycin for cytokine measurement. |
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Reaction Conditions |
10 μg/mL, 5 days |
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Applications |
T cells with allogeneic antigen-presenting cells (APCs) in vitro, and found that EX 527 significantly reduced CD4 T-cell proliferation and IFN-γ production to a comparable level of Sirt-1?/? T cells. |
| Animal experiment [2]: | |
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Animal models |
Male ZDF rats (body weight 300 ± 25 g) |
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Preparation Method |
EX-527 (5 μg/kg, twice weekly) was administered intraperitoneally (i.p.) to HFD-fed rats for ten weeks. The normal diet-fed rats received diets which were devoid of fats. Glucose levels were determined by using a glucometer. |
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Dosage form |
5 μg/kg, i.p. |
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Applications |
Two weeks after HFD-feeding, blood glucose concentration was increased significantly in HFD fed rats than that of normal diet-fed rats. However, fasting blood glucose level was dramatically decreased in the HFD-fed rats following EX 527 treatment. |
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参考文献: [1]. Daenthanasanmak A, et al. Targeting Sirt-1 controls GVHD by inhibiting T-cell allo-response and promoting Treg stability in mice. Blood. 2019 Jan 17;133(3):266-279. [2]. Kundu A, et al. EX-527 Prevents the Progression of High-Fat Diet-Induced Hepatic Steatosis and Fibrosis by Upregulating SIRT4 in Zucker Rats. Cells. 2020 Apr 29;9(5):1101. |
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