TWS119 is a highly potent and selective inhibitor of glycogen synthase kinase-3β (GSK-3) with KD value of 126nM and IC50 value of 30nM[1]. TWS119 exerts neuroprotective and regenerative effects, as well as in directing neural differentiation of stem cells and engineering immune cells[2][4][6][7].
In vitro, TWS119 (5μM) pre-incubation on mouse microglial cell line MG6 cells for 1h before stimulated with PgLPS (1μg/ml) for 48h singificantly decreased mRNA expression of TNF-α and TNF-α secretion induced by PgLPS[3]. TWS119 (20μM, 2.5μM or 5μM) was applied on NK cell line NK-92 or primary NK cells for 48h. NK cells exhibited a significantly increase in degranulation activity, expression of activating receptors, cellular cytotoxicity, and cytokine secretion when co-cultured with human multiple myeloma RPMI8226 cells for 4h[4]. TWS119 (0.5, 1.0, 2.0, 4.0, and 8.0μM) treatment on γδT cells for 72h enhanced the proliferation and survival of γδT cells via activation of the mTOR pathway, upregulation of Bcl-2 expression and inhibition of cleaved caspase-3. TWS119 treatment also upregulated the expression of perforin and granzyme B in γδT cells and enhanced the cytolytic activity of γδT cells against tumour cells[5].
In vivo, TWS119 (30mg/kg) administrated to the neonatal rats by intraperitoneal injection 20min prior to hypoxia-ischemia (HI) procedure performed a neuroprotective function at 7 days after hypoxic-ischemic brain damage via a crosstalk with Wnt/β-catenin and Notch signaling pathways at 24h after hypoxic-ischemic brain damage in neonatal rats[6]. TWS119 (30mg/kg), given intraperitoneally right after rtPA at 4h post-left middle cerebral artery in Sprague–Dawley rats, protects the blood–brain barrier and curbs hemorrhagic transformation by activating Wnt/β-catenin signaling via GSK-3β inhibition[7].
References:
[1] Ding, S. Wu T Y H, Brinker A, et al. Synthetic small molecules that control stem cell fate. 2003.Proc. Natl. Acad. Sci. USA 100, 7632–7637.
[2] Jin F J, Wu Z Z, Hu X, et al. The PI3K/Akt/GSK-3β/ROS/eIF2B pathway promotes breast cancer growth and metastasis via suppression of NK cell cytotoxicity and tumor cell susceptibility. Cancer Biol Med. 2019 Feb;16(1):38-54.
[3] Jiang M Z, Zhang X W, Yan X, et al. GSK3β is involved in promoting Alzheimer's disease pathologies following chronic systemic exposure to Porphyromonas gingivalis lipopolysaccharide in amyloid precursor proteinNL-F/NL-F knock-in mice. Brain Behav Immun. 2021 Nov:98:1-12.
[4] Ren J, Feng X M, Guo Y N, et al. GSK-3β/β-catenin pathway plays crucial roles in the regulation of NK cell cytotoxicity against myeloma cells. FASEB J. 2023 Mar;37(3):e22821.
[5] Chen Y Q, Zheng L, Aldarouish M, et al. Wnt pathway activator TWS119 enhances the proliferation and cytolytic activity of human γδT cells against colon cancer. Exp Cell Res. 2018 Jan 1;362(1):63-71.
[6] Gao L M, Yang L J, Cui H. GSK-3β inhibitor TWS119 alleviates hypoxic-ischemic brain damage via a crosstalk with Wnt and Notch signaling pathways in neonatal rats. Brain Res. 2021 Oct 1:1768:147588.
[7] Wang W, Li M C, Wang Y F, et al. GSK-3β inhibitor TWS119 attenuates rtPA-induced hemorrhagic transformation and activates the Wnt/β-catenin signaling pathway after acute ischemic stroke in rats. Mol Neurobiol. 2016 Dec;53(10):7028-7036.
TWS119是一种高亲和力且高选择性的糖原合酶激酶-3β （GSK-3）抑制剂，KD为126nM，IC50值为30nM[1]。TWS119具有神经保护与再生效应，并可用于定向诱导干细胞向神经细胞的分化以及免疫细胞工程化[2][4][6][7]。
体外实验中，TWS119 (5μM)预处理MG6小鼠小胶质细胞1小时，再加入PgLPS（1μg/ml）刺激48小时，TWS119显著抑制了PgLPS诱导的TNF-α mRNA表达及其分泌[3]。TWS119 (20、2.5或5μM)处理NK细胞系NK-92或原代NK细胞48小时后，再将NK细胞与人多发性骨髓瘤RPMI8226细胞共培养4小时，NK细胞的脱颗粒活性、激活性受体表达、对肿瘤细胞的细胞毒性和细胞因子分泌均显著增强[4]。TWS119 （0.5、1、2、4、8μM）处理γδT细胞72小时后，通过激活mTOR通路、上调Bcl-2表达并抑制cleaved caspase-3，促进γδT细胞增殖与存活；TWS119还上调γδT细胞中穿孔素和颗粒酶B的表达，增强T细胞对肿瘤细胞的细胞毒活性[5]。
体内实验中，TWS119（30mg/kg）在新生大鼠缺氧缺血（HI）手术前20分钟通过腹腔注射给药，在缺氧缺血性脑损伤后的7天通过与Wnt/β-连环蛋白和Notch信号通路的相互作用实现神经保护功能[6]。TWS119（30mg/kg）在Sprague-Dawley大鼠左侧大脑中动脉闭塞手术后4小时通过腹腔注射给药，TWS119通过抑制GSK-3β激活Wnt/β-catenin信号通路，保护血脑屏障并抑制出血性转化[7]。