State Key Laboratory of Drug Research, Shanghai Institute of Materia Medica, Chinese Academy of Sciences, Shanghai, 201203, China.
University of Chinese Academy of Sciences, Beijing, 100049, China.
Acta Pharmacol Sin. 2021 Nov;42(11):1834-1846. doi: 10.1038/s41401-020-00609-w. Epub 2021 Feb 11.
Suppression of excessive hepatic gluconeogenesis is an effective strategy for controlling hyperglycemia in type 2 diabetes (T2D). In the present study, we screened our compounds library to discover the active molecules inhibiting gluconeogenesis in primary mouse hepatocytes. We found that SL010110 (5-((4-allyl-2-methoxyphenoxy) methyl) furan-2-carboxylic acid) potently inhibited gluconeogenesis with 3 μM and 10 μM leading to a reduction of 45.5% and 67.5%, respectively. Moreover, SL010110 caused suppression of gluconeogenesis resulted from downregulating the protein level of phosphoenolpyruvate carboxykinase 1 (PEPCK1), but not from affecting the gene expressions of PEPCK, glucose-6-phosphatase, and fructose-1,6-bisphosphatase. Furthermore, SL010110 increased PEPCK1 acetylation, and promoted PEPCK1 ubiquitination and degradation. SL010110 activated p300 acetyltransferase activity in primary mouse hepatocytes. The enhanced PEPCK1 acetylation and suppressed gluconeogenesis caused by SL010110 were blocked by C646, a histone acetyltransferase p300 inhibitor, suggested that SL010110 inhibited gluconeogenesis by activating p300. SL010110 decreased NAD/NADH ratio, inhibited SIRT2 activity, and further promoted p300 acetyltransferase activation and PEPCK1 acetylation. These effects were blocked by NMN, an NAD precursor, suggested that SL010110 inhibited gluconeogenesis by inhibiting SIRT2, activating p300, and subsequently promoting PEPCK1 acetylation. In type 2 diabetic ob/ob mice, single oral dose of SL010110 (100 mg/kg) suppressed gluconeogenesis accompanied by the suppressed hepatic SIRT2 activity, increased p300 activity, enhanced PEPCK1 acetylation and degradation. Chronic oral administration of SL010110 (15 or 50 mg/kg) significantly reduced the blood glucose levels in ob/ob and db/db mice. This study reveals that SL010110 is a lead compound with a distinct mechanism of suppressing gluconeogenesis via SIRT2-p300-mediated PEPCK1 degradation and potent anti-hyperglycemic activity for the treatment of T2D.
抑制肝糖异生过度是控制 2 型糖尿病（T2D）高血糖的有效策略。在本研究中，我们从化合物库中筛选出能在原代小鼠肝细胞中抑制糖异生的活性分子。我们发现 SL010110（5-((4-烯丙基-2-甲氧基苯氧基)甲基)呋喃-2-羧酸）能强烈抑制糖异生，其 3μM 和 10μM 时的抑制率分别为 45.5%和 67.5%。此外，SL010110 导致的糖异生抑制作用是通过下调磷酸烯醇丙酮酸羧激酶 1（PEPCK1）的蛋白水平而不是影响 PEPCK、葡萄糖-6-磷酸酶和果糖-1,6-二磷酸酶的基因表达来实现的。此外，SL010110 增加了 PEPCK1 的乙酰化，促进了 PEPCK1 的泛素化和降解。SL010110 在原代小鼠肝细胞中激活了 p300 乙酰转移酶活性。C646（组蛋白乙酰转移酶 p300 抑制剂）阻断了 SL010110 引起的 PEPCK1 乙酰化增强和糖异生抑制，提示 SL010110 通过激活 p300 抑制糖异生。SL010110 降低了 NAD/NADH 比值，抑制了 SIRT2 活性，进一步促进了 p300 乙酰转移酶的激活和 PEPCK1 的乙酰化。NMN（NAD 前体）阻断了这些作用，提示 SL010110 通过抑制 SIRT2、激活 p300 进而促进 PEPCK1 乙酰化来抑制糖异生。在 2 型糖尿病 ob/ob 小鼠中，单次口服 SL010110（100mg/kg）可抑制糖异生，同时抑制肝 SIRT2 活性，增加 p300 活性，增强 PEPCK1 乙酰化和降解。慢性口服 SL010110（15 或 50mg/kg）可显著降低 ob/ob 和 db/db 小鼠的血糖水平。本研究揭示了 SL010110 是一种通过 SIRT2-p300 介导的 PEPCK1 降解来抑制糖异生的先导化合物，具有显著的抗高血糖活性，可用于治疗 T2D。
