The salt-inducible kinases (SIKs) 1 to 3, belonging to the AMPK-related kinase family, serve as master regulators orchestrating a diverse set of physiological processes such as metabolism, bone formation, immune response, oncogenesis, and cardiac rhythm. Owing to its key regulatory role, the SIK kinases have emerged as compelling targets for pharmacological intervention across a diverse set of indications. Therefore, there is interest in developing SIK inhibitors with defined selectivity profiles both to further dissect the downstream biology and for treating disease. However, despite a large pharmaceutical interest in the SIKs, experimental structures of SIK kinases are scarce. This is likely due to the challenges associated with the generation of proteins suitable for structural studies. By adopting a rational approach to construct design and protein purification, we successfully crystallized and subsequently solved the structure of SIK3 in complex with HG-9-91-01, a potent SIK inhibitor. To enable further SIK3-inhibitor complex structures we identified an antibody fragment that facilitated crystallization and enabled a robust protocol suitable for structure-based drug design. The structures reveal SIK3 in an active conformation, where the ubiquitin-associated domain is shown to provide further stabilization to this active conformation. We present four pharmacologically relevant and distinct SIK3-inhibitor complexes. These detail the key interaction for each ligand and reveal how different regions of the ATP site are engaged by the different inhibitors to achieve high affinity. Notably, the structure of SIK3 in complex with a SIK3 specific inhibitor offers insights into isoform selectivity.

2023-12-28·Zhong nan da xue xue bao. Yi xue ban = Journal of Central South University. Medical sciences

盐诱导激酶抑制剂HG-9-91-01对小鼠脓毒症相关认知功能障碍的保护作用及其机制.

Article

作者: Cui, Yanhui ; Wang, Shuang ; Wang, Xueqin

目的: 脓毒症相关认知功能障碍是脓毒症患者常见的并发症，目前病理机制不明，缺乏有效的防治手段。盐诱导激酶(salt-induced kinase，SIK)是调节代谢、免疫、炎症反应等的重要分子，与多种神经系统疾病的发生和发展相关。本研究旨在探讨SIK在脓毒症小鼠海马中的表达，以及SIK抑制剂HG-9-91-01在脓毒症相关认知功能障碍中的作用及其机制。方法: 首先，将C57BL/6小鼠随机分为对照组(Con组)和脓毒症模型组[脂多糖(lipopolysaccharide，LPS)组]。LPS组小鼠以8 mg/kg的剂量腹腔注射LPS，Con组小鼠予注射等体积生理盐水。在注射后1、3、6 d取小鼠海马组织，分别采用实时荧光定量聚合酶链反应(quantitative PCR，qPCR)和蛋白质印迹法检测SIK1、SIK2、SIK3的mRNA和蛋白质的表达水平。然后，将小鼠随机分为Con组、LPS组、SIK抑制剂组(HG组)。LPS组和HG组注射LPS构建脓毒症模型，HG组在注射LPS后的第3~6天以10 mg/kg的剂量腹腔注射HG-9-91-01，LPS组注射等体积溶媒。在注射LPS后的第7~11天进行Morris水迷宫实验(Morris water maze，MWM)以评估3组小鼠的认知功能。行为学检测后取3组小鼠的海马组织，采用qPCR检测炎症因子和小胶质细胞标志分子的mRNA表达水平，蛋白质印迹法检测诱导型一氧化氮合酶(inducible nitric oxide synthase，iNOS)、CD68、离子钙结合衔接分子1(ionized calcium binding adaptor molecule 1，Iba-1)、N-甲基-D-天冬氨酸(N-methyl-D-aspartate，NMDA)受体(NMDA receptor，NR)亚型、cAMP反应元件结合蛋白(cAMP response element-binding protein，CREB)调控的转录共激活因子1(CREB-regulated transcription coactivator 1，CRTC1)、胰岛素样生长因子1(insulin-like growth factor 1，IGF-1)的蛋白质表达水平，免疫组织化学(immunohistochemistry，IHC)检测海马CA1区、CA3区、齿状回(dentate gyrus，DG)Iba-1阳性细胞的表达，并进行Sholl分析。结果: 与Con组比较，LPS组小鼠海马组织SIK1、SIK2、SIK3的mRNA和蛋白质表达水平均上调(均P<0.05)。与Con组比较，LPS组小鼠的逃避潜伏期明显延长，目标象限停留时间百分比降低，运动速度下降(均P<0.05)；与LPS组相比，HG组小鼠的逃避潜伏期明显缩短，目标象限停留时间百分比增加(均P<0.05)。与Con组比较，LPS组海马组织炎症因子[肿瘤坏死因子α(tumor necrosis factor-α，TNF-α)、白细胞介素1β(interleukin-1β，IL-1β)、白细胞介素6(interleukin-6，IL-6)]和I型小胶质细胞标志分子iNOS、CD68的mRNA表达均上调，而II型小胶质细胞标志分子CD206和精氨酸酶-1(arginase-1，Arg-1)的mRNA表达均下调；与LPS组比较，HG组TNF-α、IL-1β、IL-6、iNOS的mRNA表达均下调，而CD206和Arg-1的mRNA表达均上调(均P<0.05)。与Con组比较，LPS组海马组织iNOS、CD68、Iba-1的蛋白质表达均上调；与LPS组比较，HG组iNOS、CD68、Iba-1的蛋白质表达均下调(均P<0.05)。与Con组比较，LPS组海马组织CA1、CA3、DG区Iba-1阳性细胞数量均增加；与LPS组比较，HG组CA1、CA3、DG区Iba-1阳性细胞数量均减少(均P<0.05)。Sholl分析结果显示：距离小胶质细胞胞体8~38 μm半径范围内，LPS组小胶质细胞突起与同心圆的交点较Con组明显减少(均P<0.05)；在距离小胶质细胞胞体14~20 μm半径范围内，HG组小胶质细胞突起与同心圆的交点较LPS组明显增多(均P<0.05)。与Con组比较，LPS组小鼠海马突触相关蛋白NR亚型NR1、NR2A、NR2B及IGF-1的蛋白质表达下调，磷酸化的CRTC1(phosphorylated CRTC1，p-CRTC1)的蛋白质表达上调；与LPS组比较，HG组小鼠海马突触相关蛋白NR1、NR2A、NR2B及IGF-1的蛋白质表达上调，p-CRTC1的蛋白质表达下调(均P<0.05)。结论: 脓毒症小鼠海马组织SIK表达上调，SIK抑制剂HG-9-91-01可改善小鼠脓毒症相关认知功能障碍，其机制可能与激活CRTC1/IGF-1通路，抑制神经炎症，增强突触可塑性有关。.

2023-04-01·Neuropharmacology

The salt-inducible kinases inhibitor HG-9-91-01 exhibits antidepressant-like actions in mice exposed to chronic unpredictable mild stress

Article

作者: Chen, Yan-Mei ; Shi, Tian-Shun ; Zhu, Bao-Lun ; Wang, Cheng-Niu ; Chen, Wei-Jia ; Li, Wei-Yu ; Liu, Jian-Feng ; Jiang, Bo ; Guan, Wei ; Fan, Hua ; Huang, Jie

Major depressive disorder is a frequently occurring neuropsychiatric disorder throughout the world. However, the limited and delayed therapeutic efficacy of monoaminergic medications has led to intensive research efforts to develop novel antidepressants. We have previously demonstrated that hippocampal salt-inducible kinase 2 (SIK2) plays a role in the pathogenesis of depression via regulating the downstream CREB-regulated transcription coactivator 1 (CRTC1)-cAMP response element-binding protein (CREB)-brain derived neurotrophic factor (BDNF) pathway. HG-9-91-01 is a potent and selective inhibitor of salt-inducible kinases (SIKs). The present study aims to explore whether HG-9-91-01 has antidepressant-like actions in male C57BL/6J mice. The chronic unpredictable mild stress (CUMS) model of depression, various behavioral tests, western blotting, co-immunoprecipitation, immunofluorescence, stereotactic infusion, and viral-mediated genetic knockdown were used together. It was found that hippocampal infusion of HG-9-91-01 induced significant antidepressant-like effects in the CUMS model, accompanied with preventing the enhancement of CUMS on the hippocampal SIK2 expression and cytoplasmic translocation of CRTC1. HG-9-91-01 treatment also reversed the decreasing effects of CUMS on the BDNF signaling cascade and adult neurogenesis in the hippocampus. Moreover, the antidepressant-like actions of HG-9-91-01 in mice required the hippocampal CRTC1-CREB-BDNF pathway. In conclusion, HG-9-91-01 has potential of being a novel antidepressant candidate.

100 项与 HG-99101 相关的药物交易

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