Feng Guo-Shuai, Zhu Cui-Ge, Li Zhuo-Ming, Wang Pan-Xia, Huang Yi, Liu Min, He Ping, Lou Lan-Lan, Chen Shao-Rui, Liu Pei-Qing
Department of Pharmacology and Toxicology, School of Pharmaceutical Sciences, Sun Yat-Sen University, Guangzhou 510006, China.
Department of Medicinal Chemistry, School of Pharmaceutical Sciences, Sun Yat-Sen University, Guangzhou 510006, China.
Acta Pharmacol Sin. 2017 May;38(5):638-650. doi: 10.1038/aps.2016.159. Epub 2017 Feb 27.
We previously identified AG-690/11026014 (6014) as a novel poly(ADP-ribose) polymerase-1 (PARP-1) inhibitor that effectively prevented angiotensin II (Ang II)-induced cardiomyocyte hypertrophy. In the present study, we reported a new synthesis route for 6014, and investigated its protective effects on Ang II-induced cardiac remodeling and cardiac dysfunction and the underlying mechanisms in mice. We designed a new synthesis route to obtain a sufficient quantity of 6014 for this in vivo study. C57BL/6J mice were infused with Ang II and treated with 6014 (10, 30, 90 mg·kg·d, ig) for 4 weeks. Then two-dimensional echocardiography was performed to assess the cardiac function and structure. Histological changes of the hearts were examined with HE staining and Masson's trichrome staining. The protein expression was evaluated by Western blot, immunohistochemistry and immunofluorescence assays. The activities of sirtuin-1 (SIRT-1) and the content of NAD+ were detected with the corresponding test kits. Treatment with 6014 dose-dependently improved cardiac function, including LVEF, CO and SV and reversed the changes of cardiac structure in Ang II-infused mice: it significantly ameliorated Ang II-induced cardiac hypertrophy evidenced by attenuating the enlargement of cardiomyocytes, decreased HW/BW and LVW/BW, and decreased expression of hypertrophic markers ANF, BNP and β-MHC; it also prevented Ang II-induced cardiac fibrosis, as implied by the decrease in excess accumulation of extracellular matrix (ECM) components collagen I, collagen III and FN. Further studies revealed that treatment with 6014 did not affect the expression levels of PARP-1, but dose-dependently inhibited the activity of PARP-1 and subsequently restored the activity of SIRT-1 in heart tissues due to the decreased consumption of NAD+ and attenuated Poly-ADP-ribosylation (PARylation) of SIRT-1. In conclusion, the novel PARP-1 inhibitor 6014 effectively protects mice against AngII-induced cardiac remodeling and improves cardiac function. Thus, 6014 might be a potential therapeutic agent for heart diseases..
我们之前鉴定出AG-690/11026014（6014）是一种新型的聚（ADP-核糖）聚合酶-1（PARP-1）抑制剂，它能有效预防血管紧张素II（Ang II）诱导的心肌细胞肥大。在本研究中，我们报道了6014的一种新合成路线，并研究了其对Ang II诱导的心脏重塑和心脏功能障碍的保护作用以及在小鼠体内的潜在机制。我们设计了一种新的合成路线以获得足够量的6014用于此项体内研究。将C57BL/6J小鼠用Ang II灌注，并给予6014（10、30、90 mg·kg·d，灌胃）处理4周。然后进行二维超声心动图以评估心脏功能和结构。用HE染色和Masson三色染色检查心脏的组织学变化。通过蛋白质印迹、免疫组织化学和免疫荧光分析评估蛋白质表达。用相应的检测试剂盒检测沉默调节蛋白-1（SIRT-1）的活性和NAD+的含量。6014处理剂量依赖性地改善了心脏功能，包括左心室射血分数（LVEF）、心输出量（CO）和每搏输出量（SV），并逆转了Ang II灌注小鼠的心脏结构变化：它显著改善了Ang II诱导的心肌肥大，表现为心肌细胞增大减轻、心脏重量/体重（HW/BW）和左心室重量/体重（LVW/BW）降低以及肥大标志物心房钠尿肽（ANF）、脑钠肽（BNP）和β-肌球蛋白重链（β-MHC）表达减少；它还预防了Ang II诱导的心脏纤维化，这表现为细胞外基质（ECM）成分I型胶原、III型胶原和纤连蛋白（FN）过量积累减少。进一步的研究表明，6014处理不影响PARP-1的表达水平，但剂量依赖性地抑制PARP-1的活性，随后由于NAD+消耗减少和SIRT-1的聚ADP核糖基化（PARylation）减弱而恢复了心脏组织中SIRT-1的活性状态。总之，新型PARP-1抑制剂6014能有效保护小鼠免受Ang II诱导的心脏重塑并改善心脏功能。因此，6014可能是一种潜在的心脏病治疗药物。
