1] Graduate School of Pharmaceutical Sciences, Osaka University, Suita, Japan [2] Department of Molecular Innovation in Lipidology, National Cerebral and Cardiovascular Center Research Institute, Suita, Japan.
Mol Ther Nucleic Acids. 2012 May 15;1(5):e22. doi: 10.1038/mtna.2012.16.
Recent findings in molecular biology implicate the involvement of proprotein convertase subtilisin/kexin type 9 (PCSK9) in low-density lipoprotein receptor (LDLR) protein regulation. The cholesterol-lowering potential of anti-PCSK9 antisense oligonucleotides (AONs) modified with bridged nucleic acids (BNA-AONs) including 2',4'-BNA (also called as locked nucleic acid (LNA)) and 2',4'-BNA(NC) chemistries were demonstrated both in vitro and in vivo. An in vitro transfection study revealed that all of the BNA-AONs induce dose-dependent reductions in PCSK9 messenger RNA (mRNA) levels concomitantly with increases in LDLR protein levels. BNA-AONs were administered to atherogenic diet-fed C57BL/6J mice twice weekly for 6 weeks; 2',4'-BNA-AON that targeted murine PCSK9 induced a dose-dependent reduction in hepatic PCSK9 mRNA and LDL cholesterol (LDL-C); the 43% reduction of serum LDL-C was achieved at a dose of 20 mg/kg/injection with only moderate increases in toxicological indicators. In addition, the serum high-density lipoprotein cholesterol (HDL-C) levels increased. These results support antisense inhibition of PCSK9 as a potential therapeutic approach. When compared with 2',4'-BNA-AON, 2',4'-BNA(NC)-AON showed an earlier LDL-C-lowering effect and was more tolerable in mice. Our results validate the optimization of 2',4'-BNA(NC)-based anti-PCSK9 antisense molecules to produce a promising therapeutic agent for the treatment of hypercholesterolemia.
最近的分子生物学研究结果表明，前蛋白转化酶枯草溶菌素/柯萨奇蛋白酶 9（PCSK9）参与了低密度脂蛋白受体（LDLR）蛋白的调节。用桥接核酸（BNA）修饰的抗 PCSK9 反义寡核苷酸（AON），包括 2'，4'-BNA（也称为锁核酸（LNA））和 2'，4'-BNA（NC）化学，在体外和体内都显示出降低胆固醇的潜力。一项体外转染研究表明，所有的 BNA-AON 都能诱导 PCSK9 信使 RNA（mRNA）水平的剂量依赖性降低，同时 LDLR 蛋白水平升高。BNA-AON 每周两次给喂食动脉粥样硬化饮食的 C57BL/6J 小鼠注射 6 周；靶向鼠 PCSK9 的 2'，4'-BNA-AON 诱导肝 PCSK9 mRNA 和 LDL 胆固醇（LDL-C）的剂量依赖性降低；在 20mg/kg/注射剂量下，血清 LDL-C 降低 43%，而毒性指标仅略有升高。此外，血清高密度脂蛋白胆固醇（HDL-C）水平升高。这些结果支持反义抑制 PCSK9 作为一种潜在的治疗方法。与 2'，4'-BNA-AON 相比，2'，4'-BNA（NC）-AON 显示出更早的 LDL-C 降低作用，在小鼠中更耐受。我们的结果验证了基于 2'，4'-BNA（NC）的抗 PCSK9 反义分子的优化，以产生一种有前途的治疗高胆固醇血症的治疗剂。
