JNK-IN-8 is the first irreversible JNK inhibitor that acts on JNK1, JNK2, and JNK3 with high specificity, with IC50 values of 4.7 nM, 18.7 nM, and 1 nM in the A375 cell line, respectively[1]. JNK-IN-8 forms a covalent bond with the conserved cysteine residue of JNK 1/2/3, causing a conformational change in the activation loop and blocking substrate binding, thereby inhibiting the activity of JNK 1/2/3[1]. JNK 1, 2, and 3 are part of the mitogen-activated protein kinase (MAPK) family, capable of phosphorylating the Ser 63 and Ser 73 residues of c-Jun, responding to stress stimuli such as cytokines and heat shock, and involved in T cell differentiation and apoptosis processes.
In vitro, JNK-IN-8 inhibits c-Jun phosphorylation in HeLa and A375 cells, with EC50 values of 486 nM and 338 nM, respectively[1]. JNK-IN-8 also shows significant selectivity in HEK 293 cells[1]. Additionally, JNK-IN-8 (10 mM) exhibits anti-inflammatory effects, reducing microglial activation and the expression of IL-6, IL-1β, and TNF-α[2].
In vivo, JNK-IN-8 (20 mg/kg; i.p.) treated rats with middle cerebral artery occlusion show significant improvements in spatial learning and sensorimotor function recovery[2]. Administration of JNK-IN-8 (3µg/µL) into the lateral ventricles of male KM mice 24 h before brain injury significantly reduces neuronal apoptosis after brain injury[3].
References:
[1] Zhang T, Inesta-Vaquera F, et al, Discovery of potent and selective covalent inhibitors of JNK. Chemical Biology. 2012, 19(1):140-154.
[2] Geng W , Tang H , Dai Q ,et al. JNK-IN-8, a c-Jun N-terminal kinase inhibitor, improves functional recovery through suppressing neuroinflammation in ischemic stroke[J].Cold Spring Harbor Laboratory, 2018.
[3] Li D1, Liu N, et al, Protective effect of resveratrol against nigrostriatal pathway injury in striatum via JNK pathway. Brain Res. 2017 Jan 1;1654(Pt A):1-8.
JNK-IN-8 是第一个不可逆的JNK抑制剂,作用于JNK1,JNK2和JNK3,具有高度特异性,在A375细胞系中IC50分别为4.7 nM,18.7 nM和1 nM[1]。JNK-IN-8与JNK 1/2/3的保守半胱氨酸残基形成共价键,导致激活环的构象变化,阻断底物结合,从而抑制JNK 1/2/3的活性[1]。JNK 1、2和3属于丝裂原活化蛋白激酶(mitogen-activated protein kinase,MAPK)家族,能够磷酸化c-Jun的Ser 63和Ser 73残基,对细胞因子和热休克等应激刺激有应答,参与T细胞分化和细胞凋亡过程。
在体外,JNK-IN-8 抑制 HeLa和A375细胞中的c-Jun磷酸化,EC50分别为 486 nM和338 nM[1]。JNK-IN-8在HEK 293细胞中还表现出显著的选择性[1]。JNK-IN-8 (10 mM)还发挥抗炎作用,使小胶质细胞活化减少以及 IL-6、IL-1β和TNF-α 表达减少[2]。
在体内,JNK-IN-8 (20 mg/kg; i.p.)治疗的大脑中动脉闭塞大鼠在空间学习方面表现出显着改善,并且感觉运动功能恢复[2]。JNK-IN-8在雄性KM小鼠脑损伤前24 h向侧脑室注射3µg/µL 能显著减少脑损伤后神经元凋亡[3]。