Inflammation in the central nervous system (CNS) is observed in many neurological disorders. Nitric oxide-soluble guanylate cyclase-cyclic guanosine monophosphate (NO–sGC–cGMP) signaling plays an essential role in modulating neuroinflammation. CYR119 is a CNS-penetrant sGC stimulator that amplifies endogenous NO–sGC–cGMP signaling. We evaluated target engagement and the effects of CYR119 on markers of neuroinflammation in vitro in mouse microglial cells and in vivo in quinolinic acid (QA)-induced and high-fat diet-induced rodent neuroinflammation models. Target engagement was verified in human embryonic kidney (HEK) cells, rat primary neurons, mouse SIM-A9 cells, and in rats by measuring changes in cGMP and downstream targets of sGC signaling [phosphorylated vasodilator-stimulated phosphoprotein (pVASP), phosphorylated cAMP-response element binding (pCREB)]. In SIM-A9 cells stimulated with lipopolysaccharides (LPS), markers of inflammation were measured when cells were treated with or without CYR119. In rats, microinjections of QA and vehicle were administered into the right and left hemispheres of striatum, respectively, and then rats were dosed daily with either CYR119 (10 mg/kg) or vehicle for 7 days. The activation of microglia [ionized calcium binding adaptor molecule 1 (Iba1)] and astrocytes [glial fibrillary acidic protein (GFAP)] was measured by immunohistochemistry. Diet-induced obese (DIO) mice were treated daily with CYR119 (10 mg/kg) for 6 weeks, after which inflammatory genetic markers were analyzed in the prefrontal cortex. In vitro, CYR119 synergized with exogenous NO to increase the production of cGMP in HEK cells and in primary rat neuronal cell cultures. In primary neurons, CYR119 stimulated sGC, resulting in accumulation of cGMP and phosphorylation of CREB, likely through the activation of protein kinase G (PKG). CYR119 attenuated LPS-induced elevation of interleukin 6 (IL-6) and tumor necrosis factor (TNF) in mouse microglial cells. Following oral dosing in rats, CYR119 crossed the blood–brain barrier (BBB) and stimulated an increase in cGMP levels in the cerebral spinal fluid (CSF). In addition, levels of proinflammatory markers associated with QA administration or high-fat diet feeding were lower in rodents treated with CYR119 than in those treated with vehicle. These data suggest that sGC stimulation could provide neuroprotective effects by attenuating inflammatory responses in nonclinical models of neuroinflammation.

中文翻译：

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CNS 渗透可溶性鸟苷酸环化酶刺激剂 CYR119 减弱中枢神经系统炎症标志物

在许多神经系统疾病中观察到中枢神经系统 (CNS) 的炎症。一氧化氮可溶性鸟苷酸环化酶-环鸟苷一磷酸 (NO-sGC-cGMP) 信号传导在调节神经炎症中起重要作用。CYR119 是一种 CNS 渗透性 sGC 刺激剂，可放大内源性 NO-sGC-cGMP 信号传导。我们评估了 CYR119 对体外小鼠小胶质细胞和体内喹啉酸 (QA) 诱导和高脂饮食诱导的啮齿动物神经炎症模型的神经炎症标志物的靶向参与和影响。通过测量 cGMP 和 sGC 信号下游靶标 [磷酸化血管扩张剂刺激的磷蛋白 (pVASP)、磷酸化 cAMP 反应元件结合 (pCREB)]。在用脂多糖 (LPS) 刺激的 SIM-A9 细胞中，在用或不用 CYR119 处理细胞时测量炎症标志物。在大鼠中，分别将 QA 和载体微量注射到纹状体的右半球和左半球，然后每天给大鼠服用 CYR119 (10 mg/kg) 或载体，持续 7 天。通过免疫组织化学测量小胶质细胞 [离子化钙结合衔接分子 1 (Iba1)] 和星形胶质细胞 [胶质纤维酸性蛋白 (GFAP)] 的激活。饮食诱导的肥胖 (DIO) 小鼠每天用 CYR119 (10 mg/kg) 治疗 6 周，之后在前额皮质中分析炎症遗传标记。体外，CYR119 与外源 NO 协同作用以增加 HEK 细胞和原代大鼠神经元细胞培养物中 cGMP 的产生。在原代神经元中，CYR119 刺激 sGC，导致 cGMP 的积累和 CREB ​​的磷酸化，可能是通过激活蛋白激酶 G (PKG)。CYR119 减弱 LPS 诱导的小鼠小胶质细胞中白细胞介素 6 (IL-6) 和肿瘤坏死因子 (TNF) 的升高。大鼠口服给药后，CYR119 穿过血脑屏障 (BBB) 并刺激脑脊液 (CSF) 中 cGMP 水平的增加。此外，用 CYR119 治疗的啮齿动物中与 QA 给药或高脂饮食喂养相关的促炎标志物水平低于用载体治疗的啮齿动物。