Mitochondria-targeting Novel Cationic Hydrazone Antioxidants for the Treatment of Preeclampsia

摘要 子痫前期(PE)是妊娠期一种常见的危及生命的并发症，其特征是高血 压力使母亲有患子痫和肾功能障碍的风险。PE影响约1%-5%的 怀孕是孕产妇和胎儿死亡的一个主要原因。私募股权投资的特点是 滋养层细胞的侵袭和螺旋动脉的异常重构；以及 血管生成/抗血管生成失衡。异常的胎盘很可能会导致胎盘减少- 母体血流和氧化应激增加，导致功能受损。没有明确的 因此，PE的有效治疗是一个尚未得到满足的医学需求。的主要目标是 这项建议是为了开发新型的以线粒体为靶点的阳离子有机氟肼类化合物。 先兆子痫的抗氧化治疗。三个关键的结构特征将结合在一起，以有效 抗氧化剂：(I)用于提高抗氧化潜力的肼核心，(Ii)掺入氟用于 增加亲油性和膜的通透性，以及(Iii)季铵盐的引入 (Qass)用于通过线粒体膜输送亲脂性阳离子。基于我们的 初步数据和计算机评估的结果，我们将设计和合成含有QAS的 将对有机氟抗氧化剂进行体外抗氧化活性初步筛选 在生化分析中。将显示出显著清除自由基性能的化合物将是 在基于细胞和体内的测试中进行评估。人原代滋养层细胞(正常妊娠或PE妊娠) 将测试所建议的抗氧化剂在化学上增强细胞氧化还原功能是否会 减少(I)细胞损伤，(Ii)线粒体应激，(Iii)HIF1α的产生，以及(Iv)下游抗血管生成 对缺氧-复氧的反应。同样的检测也将在人类内皮细胞中进行。 暴露在缺氧-复氧环境中。此外，长期怀孕的小鼠将受到缺氧的影响， 诱导PE的主要特征(高血压、蛋白尿和氧化应激)，并给予西药治疗 将对合成的抗氧化剂和小鼠进行血压、肾功能、组织学评估 损伤、3-硝基酪氨酸(3-NT)组织免疫反应、炎症标志物和血浆生物标志物。 最后，将使用质谱学(MS)和核磁共振(核磁共振)谱来 绘制细胞和组织中新的阳离子肼的存在图。这些研究可能会导致 开发新的有效抗氧化剂化合物，拮抗ROS/RNS，防止 激活HIF1通路，从而改善血管生成平衡，降低全身 体育运动的影响。在教育/培训方面，这项工作将提供一个多环境 以及对将参与项目每一步的本科生进行跨学科培训 并接触到两个不同机构的研究氛围。

SUMMARY Preeclampsia (PE) is a common, life-threatening complication in pregnancy, characterized by high blood pressure putting the mother at risk of eclampsia and kidney dysfunction. PE affects about 1–5% of pregnancies and a major cause of maternal and fetal deaths. The hallmarks of PE are the decrease in trophoblast invasion and abnormal remodeling of the spiral arteries; as well as an angiogenic/antiangiogenic imbalance. The abnormal placentation will, most likely, cause reduced feto- maternal blood flow and increased oxidative stress resulting in compromised function. There is no clear pathogenesis or cure, thus an effective treatment for PE is an unmet medical need. The primary goal of this proposal is to develop novel cationic organofluorine hydrazones for mitochondria-targeted antioxidant therapy in preeclampsia. Three key structural features will be combined to effective antioxidants: (i) a hydrazone core for improved antioxidant potential, (ii) fluorine incorporation for increased lipophilicity and membrane permeability, and (iii) introduction of quaternary ammonium salts (QASs) for the delivery of the lipophilic cation through the mitochondrial membrane. Based on our preliminary data and the results of in silico evaluations, we will design and synthesize QAS-containing organofluorine antioxidants that will be subjected to primary screening for their in vitro antioxidant activity in biochemical assays. The compounds that will show significant radical scavenging properties will be evaluated in cell-based and in vivo assays. Human primary trophoblast cells (normal or PE pregnancy) will be tested whether augmenting cell-redox function chemically by the proposed antioxidants will reduce (i) cell injury, (ii) mitochondrial stress, (iii) HIF1α production, and (iv) downstream anti-angiogenic response to hypoxia-reoxygenation. The same assays will also be carried out in human endothelial cells exposed to hypoxia-reoxygenation. In addition, time-pregnant mice will be subjected to hypoxia, to induce the main features of PE, (hypertension, proteinuria and oxidative stress), and treated with the synthetic antioxidants and the mice will be evaluated for blood pressure, renal function, histologic damage, 3-nitrotyrosine (3-NT) tissue immunoreactivity, inflammation markers and plasma biomarkers. Finally, mass spectrometry (MS) and nuclear magnetic resonance (NMR) spectroscopy will be used to map the presence of the new cationic hydrazones in cells and tissues. These studies may lead to the development of new effective antioxidant compounds, which antagonize ROS/RNS that prevent the activation of the HIF1 pathway and therefore improve angiogenic balance and reduce the systemic effects of PE. Regarding the educational/training aspects, this work will provide a multi-environmental and interdisciplinary training to undergraduate students who will participate in each step of the project and be exposed to the research atmosphere at two different institutions.