一种 PCSK9 转录的小分子抑制剂通过调节 FoxO1/3 和 HNF1α 改善动脉粥样硬化。
A small-molecule inhibitor of PCSK9 transcription ameliorates atherosclerosis through the modulation of FoxO1/3 and HNF1α.
机构信息
NHC Key Laboratory of Biotechnology of Antibiotics, Institute of Medicinal Biotechnology, Chinese Academy of Medical Sciences & Peking Union Medical College, No. 1 Tiantan xili, Beijing 100050, China.
NHC Key Laboratory of Biotechnology of Antibiotics, Institute of Medicinal Biotechnology, Chinese Academy of Medical Sciences & Peking Union Medical College, No. 1 Tiantan xili, Beijing 100050, China; CAMS Key Laboratory of Synthetic Biology for Drug Innovation, Institute of Medicinal Biotechnology, Chinese Academy of Medical Sciences & Peking Union Medical College, No. 1 Tiantan xili, Beijing 100050, China.
出版信息
EBioMedicine. 2020 Feb;52:102650. doi: 10.1016/j.ebiom.2020.102650. Epub 2020 Feb 12.
BACKGROUND
Proprotein convertase subtilisin/kexin type 9 (PCSK9) is a secreted protein that down-regulates hepatic low-density lipoprotein receptor (LDLR) by binding and shuttling LDLR to lysosomes for degradation. The development of therapy that inhibits PCSK9 has attracted considerable attention for the management of cardiovascular disease risk. However, only monoclonal antibodies of PCSK9 have reached the clinic use. Oral administration of small-molecule transcriptional inhibitors has the potential to become a therapeutic option.
METHODS
Here, we developed a cell-based small molecule screening platform to identify transcriptional inhibitors of PCSK9. Through high-throughput screening and a series of evaluation, we found several active compounds. After detailed investigation on the pharmacological effect and molecular mechanistic characterization, 7030B-C5 was identified as a potential small-molecule PCSK9 inhibitor.
FINDINGS
Our data showed that 7030B-C5 down-regulated PCSK9 expression and increased the total cellular LDLR protein and its mediated LDL-C uptake by HepG2 cells. In both C57BL/6 J and ApoE KO mice, oral administration of 7030B-C5 reduced hepatic and plasma PCSK9 level and increased hepatic LDLR expression. Most importantly, 7030B-C5 inhibited lesions in en face aortas and aortic root in ApoE KO mice with a slight amelioration of lipid profiles. We further provide evidences suggesting that transcriptional regulation of PCSK9 by 7030B-C5 mostly depend on the transcriptional factor HNF1α and FoxO3. Furthermore, FoxO1 was found to play an important role in 7030B-C5 mediated integration of hepatic glucose and lipid metabolism.
INTERPRETATION
7030B-C5 with potential suppressive effect of PCSK9 expression may serve as a promising lead compound for drug development of cholesterol/glucose homeostasis and cardiovascular disease therapy. FUND: This work was supported by grants from the National Natural Science Foundation of China (81473214, 81402929, and 81621064), the Drug Innovation Major Project of China (2018ZX09711001-003-006, 2018ZX09711001-007 and 2018ZX09735001-002), CAMS Innovation Fund for Medical Sciences (2016-I2M-2-002, 2016-I2M-1-011 and 2017-I2M-1-008), Beijing Natural Science Foundation (7162129).
背景
前蛋白转化酶枯草溶菌素 9（PCSK9）是一种分泌蛋白，通过与 LDLR 结合并将 LDLR 转运至溶酶体进行降解，从而下调肝脏 LDLR。抑制 PCSK9 的治疗方法的发展引起了人们对心血管疾病风险管理的极大关注。然而，只有 PCSK9 的单克隆抗体已达到临床应用。小分子转录抑制剂的口服给药有可能成为一种治疗选择。
方法
在这里，我们开发了一种基于细胞的小分子筛选平台，以鉴定 PCSK9 的转录抑制剂。通过高通量筛选和一系列评估，我们发现了几种活性化合物。在对药理作用和分子机制特征进行详细研究后，确定 7030B-C5 为潜在的小分子 PCSK9 抑制剂。
结果
我们的数据表明，7030B-C5 下调 PCSK9 的表达，并增加 HepG2 细胞中总细胞 LDLR 蛋白及其介导的 LDL-C 摄取。在 C57BL/6J 和 ApoE KO 小鼠中，7030B-C5 的口服给药均降低了肝和血浆 PCSK9 水平，并增加了肝 LDLR 的表达。最重要的是，7030B-C5 抑制了 ApoE KO 小鼠主动脉正面和主动脉根部的病变，并略微改善了脂质谱。我们进一步提供的证据表明，7030B-C5 对 PCSK9 的转录调节主要依赖于转录因子 HNF1α 和 FoxO3。此外，发现 FoxO1 在 7030B-C5 介导的肝葡萄糖和脂质代谢整合中起重要作用。
解释
具有抑制 PCSK9 表达潜力的 7030B-C5 可能成为胆固醇/葡萄糖稳态和心血管疾病治疗药物开发的有前途的先导化合物。
资金
本工作得到了国家自然科学基金（81473214、81402929 和 81621064）、中国药物创新重大项目（2018ZX09711001-003-006、2018ZX09711001-007 和 2018ZX09735001-002）、中国医学科学院医学与健康科技创新工程（2016-I2M-2-002、2016-I2M-1-011 和 2017-I2M-1-008）、北京市自然科学基金（7162129）的资助。
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