Discovery of 5-Benzylidene-2-phenyl-1,3-dioxane-4,6-diones as Highly Potent and Selective SIRT1 Inhibitors
Discovery of 5-Benzylidene-2-phenyl-1,3-dioxane-4,6-diones as Highly Potent and Selective SIRT1 Inhibitors
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DOI：
10.1021/acsmedchemlett.0c00559
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发表时间：
2021-03-01
影响因子：
4.2
通讯作者：
Liu, Hong
中科院分区：
文献类型：
--
作者：
Li, Chunpu;Hu, Sha-Sha;Liu, Hong
SIRT 1是沉默调节蛋白家族的成员，催化蛋白质的脱乙酰化，将NAD（+）转化为烟酰胺和2 '-O-乙酰基-ADP-核糖。选择性SIRT 1/2抑制剂在结直肠癌、前列腺癌和髓性白血病的化疗中具有潜在的应用价值。在这里，我们确定了新的SIRT 1抑制剂与5-亚苄基-2-苯基-1，3-二氧六环-4，6-二酮的支架。最有效的抑制剂12 n显示出460 nM的IC 50，对SIRT 1的选择性分别是SIRT 2、SIRT 3和SIRTS的113.5倍、254.3倍和10.83倍。它不影响SIRT 6的活性。为阐明其抑制机制，我们通过酶动力学分析确定了该抑制剂的抑制类型，结果表明该抑制剂对乙酰肽具有竞争性，对NAD（+）具有非竞争性。通过分子对接方法研究了SIRT 1中抑制剂的相互作用，并通过抑制剂的构效关系分析和SIRT 1的定点突变进行了验证。与体外试验一致，抑制剂在细胞中以浓度依赖性方式增加p53的乙酰化水平。
SIRT1, a member of the sirtuin family, catalyzes the deacetylation of proteins with the transformation of NAD(+) into nicotinamide and 2'-O-acetyl-ADP-ribose. Selective SIRT1/2 inhibitors have potential application in the chemotherapy of colorectal carcinoma, prostate cancer, and myelogenous leukemia. Here we identified novel SIRT1 inhibitors with the scaffold of 5-benzylidene-2-phenyl-1,3-dioxane-4,6-dione. The most potent inhibitor 12n displayed an IC50 of 460 nM and a selectivity for SIRT1 over SIRT2, SIRT3, and SIRTS of 113.5-, 254.3-, and 10.83-fold, respectively. It did not affect the activity of SIRT6. To elucidate the inhibitory mechanism, we determined the inhibition type of the inhibitor by enzyme kinetic analysis, showing that the inhibitor was competitive to the acetyl peptide and noncompetitive to NAD(+). Further, the interaction of the inhibitor in SIRT1 was studied by using molecular docking, which was validated by the structure-activity relationship analysis of the inhibitors and the site-directed mutagenesis of SIRT1. Consistent with the in vitro assays, the inhibitors increased the acetylation level of p53 in a concentration-dependent manner in cells.