Cell permeant peptidomimetics to prevent delayed vasospasm and neurological deficits after subarachnoid hemorrhage

项目总结 颅内动脉瘤破裂引起的蛛网膜下腔出血(SAH)导致延迟 导致神经缺血(中风)的血管痉挛。总的发病率(严重的神经功能缺陷 10-20%的幸存者)和死亡率(50%)很高，这种疾病影响的是相对年轻的成年人 人口。预防蛛网膜下腔出血后迟发性血管痉挛和神经缺血的治疗选择是 目前仅限于血流动力学优化和尼莫地平，这两种药物的临床疗效微乎其微。 因此，SAH后迟发性血管痉挛的治疗对孤儿来说是一个未得到满足的临床需求。 具有严重临床后果的人群。 使用现有的血管扩张剂治疗SAH诱导的血管痉挛的尝试经常失败，因为 低血压(导致脑灌注量减少)和脑血管系统对 一氧化氮(NO)依赖的信号通路的激活。5 NO信号调节血管 平滑肌(VSM)的松弛和脑血流量的调节。受损害的反应 脑血管到血管扩张剂，即SAH后血管松弛受损，可能是由于下调调节所致 SAH后NO通路中信号元件的变化。这项调查的假设是 用合理设计的、细胞意向的磷酸多肽模拟下游效应物进行治疗 NO途径的蛋白将绕过下调的信号元件，恢复血管松弛，并 预防蛛网膜下腔出血后迟发性血管痉挛。这种方法更有针对性，也更符合化学计量比 激活或抑制受体或酶的方法。此外，这种方法尤其是 在预防全身低血压和优化脑血管扩张的SAH中很有用 派拉蒙。 CGMP依赖蛋白底物的一族细胞活性磷酸肽类似物 激酶(PKG)和一种调节VSM松弛的肌动蛋白相关蛋白被合理设计 并合成了。三个候选多肽在体外被证明直接松弛完整的VSM 生物活性检测。序列最短(记为VP3)且生物活性最强的多肽 被选为测定体内疗效的最佳多肽。

PROJECT SUMMARY Subarachnoid hemorrhage (SAH) due to rupture of an intracranial aneurysm leads to delayed vasospasm resulting in neuroischemia (stroke). The overall morbidity (profound neurologic deficit in 10-20% of survivors) and mortality (50%) are high, and the disease affects a relatively young adult population. Therapeutic options to prevent delayed vasospasm and neuroischemia after SAH are currently limited to hemodynamic optimization and nimodipine, which have marginal clinical efficacy. Thus, treatment of delayed vasospasm after SAH represents an unmet clinical need in an orphan population with severe clinical consequences. Attempts to treat SAH-induced vasospasm with existing vasodilators often fail because of systemic hypotension (leading to decreased cerebral perfusion) and a cerebral vasculature that is refractory to activation of nitric oxide (NO)-dependent signaling pathways.5 NO signaling modulates vascular smooth muscle (VSM) relaxation and regulation of cerebral blood flow. The impaired response of cerebral vessels to vasodilators, i.e. impaired vasorelaxation after SAH, is likely due to down regulation of the signaling elements in the NO pathway after SAH. The hypothesis of this investigation is that treatment with a rationally designed, cell permeant phosphopeptide mimetic of a downstream effector protein of the NO pathway will bypass downregulated signaling elements, restore vasorelaxation, and prevent delayed vasospasm after SAH. This approach is more targeted and stoichiometric than approaches that activate or inhibit receptors or enzymes. In addition, this approach is particularly useful in SAH where preventing systemic hypotension and optimizing cerebral vasodilation is paramount. A family of cell permeant phosphopeptide analogues of a substrate of cGMP-dependent Protein Kinase (PKG), and an actin-associated protein that modulates VSM relaxation, were rationally designed and synthesized. Three candidate peptides were demonstrated to directly relax intact VSM in ex vivo bioactivity assays. The peptide with the shortest sequence (denoted as VP3) and strongest bioactivity was chosen as the optimal peptide for use to determine in vivo efficacy.