GSK-872 is a RIPK3 inhibitor. GSK-872 decreases the RIPK3-mediated necroptosis and subsequent cytoplasmic translocation and expression of HMGB1, as well as ameliorates brain edema and neurological deficits in early brain injury[5]. GSK-872 bound RIP3 kinase domain with high affinity (IC50 = 1.8 nM) and inhibited kinase activity (IC50 = 1.3 nM) [1]. GSK-872 prevented virus-induced necrosis, a pathway dependent on DAI-RIP3 complex formation,GSK-872 blocked TLR3-induced necrosis induced in fibroblasts by poly(I:C) in the presence of Z-VAD-fmk, Both virus- and TLR3-induced necrosis proceed independently of RIP1 kinase inhibition by Nec-1 but sensitive to inhibition by GSK-872 [2,3,4]. Pharmacological inhibitor GSK-872 enhanced insulin signaling in vitro and in vivo, which contributing to improve insulin sensitivity[9].
When evaluated in cell culture using human HT-29 cells, GSK-872 bind the kinase domain and inhibit kinase activity with high specificity, targeting a broader range of pronecrotic stimuli [1]. RIP3i compounds GSK-872 blocked TNF-induced necroptosis in a concentration-dependent manner . In cell-based assays, there was a 100- to 1,000-fold shift in the IC50 compared to the cell-free biochemical assays. GSK-872 blocked necroptosis in primary human neutrophils isolated from whole blood，and blocked necroptosis in mouse cells. Mouse bone-marrow-derived macrophages (BMDMs) or thioglycolate-elicited peritoneal macrophages (PECs), as well as 3T3SA fibroblasts, were also protected by GSK-872 concentrations .
GSK-872 significantly reduced brain edema and improved neurological function in SAH rats, and reduced the number of necrotic cells. The exact mechanism of GSK-872 induced neuroprotective effect against SAH was identified[6,7].Inhibiting of RIPK3 by GSK-872 could attenuate RIPK3-dependent necroptosis, decrease brain edema, and improve neurological function after SAH. GSK-872 also improves hepatic steatosis and liver injury in mice fed with HFCD after CIH exposure[8].
参考文献:
[1]: Mandal P, Berger SB, et, al. RIP3 induces apoptosis independent of pronecrotic kinase activity. Mol Cell. 2014 Nov 20;56(4):481-95. doi: 10.1016/j.molcel.2014.10.021. Epub 2014 Nov 20. PMID: 25459880; PMCID: PMC4512186.
[2]:Kaiser WJ, Sridharan H, et, al. Toll-like receptor 3-mediated necrosis via TRIF, RIP3, and MLKL. J Biol Chem. 2013 Oct 25;288(43):31268-79. doi: 10.1074/jbc.M113.462341. Epub 2013 Sep 9. PMID: 24019532; PMCID: PMC3829437.
[3]: Arora D, Siddiqui MH, et, al. Deltamethrin induced RIPK3-mediated caspase-independent non-apoptotic cell death in rat primary hepatocytes. Biochem Biophys Res Commun. 2016 Oct 14;479(2):217-223. doi:
[4]: He S, Liang Y, Shao F, Wang X. Toll-like receptors activate programmed necrosis in macrophages through a receptor-interacting kinase-3-mediated pathway. Proc Natl Acad Sci U S A. 2011 Dec 13;108(50):20054-9. doi: 10.1073/pnas.1116302108. Epub 2011 Nov 28. PMID: 22123964; PMCID: PMC3250173.
[5]: Chen T, Pan H, et, al. Inhibiting of RIPK3 attenuates early brain injury following subarachnoid hemorrhage: Possibly through alleviating necroptosis. Biomed Pharmacother. 2018 Nov;107:563-570. doi: 10.1016/j.biopha.2018.08.056. Epub 2018 Aug 14. PMID: 30114640.
[6]: Chen S, Lv X, et, al.RIPK1/RIPK3/MLKL-mediated necroptosis contributes to compression-induced rat nucleus pulposus cells death. Apoptosis. 2017 May;22(5):626-638. doi: 10.1007/s10495-017-1358-2. PMID: 28289909.
[7]: Liu T, Zhao DX, et, al. Therapeutic hypothermia attenuates tissue damage and cytokine expression after traumatic brain injury by inhibiting necroptosis in the rat. Sci Rep. 2016 Apr 15;6:24547. doi: 10.1038/srep24547. PMID: 27080932; PMCID: PMC4832230.
[8]: Zhang H, Zhou L, Z et, al. Intermittent hypoxia aggravates non-alcoholic fatty liver disease via RIPK3-dependent necroptosis-modulated Nrf2/NFκB signaling pathway. Life Sci. 2021 Nov 15;285:119963. doi: 10.1016/j.lfs.2021.119963. Epub 2021 Sep 16. PMID: 34536498.
[9]:Xu H, Du X, et, al. The pseudokinase MLKL regulates hepatic insulin sensitivity independently of inflammation. Mol Metab. 2019 May;23:14-23. doi: 10.1016/j.molmet.2019.02.003. Epub 2019 Feb 20. PMID: 30837196; PMCID: PMC6480316.
GSK-872 是一种 RIPK3 抑制剂。 GSK-872 减少 RIPK3 介导的坏死性凋亡和随后的 HMGB1 细胞质易位和表达，并改善早期脑损伤的脑水肿和神经功能缺损[5]。 GSK-872 以高亲和力 (IC50 = 1.8 nM) 结合 RIP3 激酶结构域并抑制激酶活性 (IC50 = 1.3 nM) [1]。 GSK-872 阻止了病毒诱导的坏死，一种依赖于 DAI-RIP3 复合物形成的途径，GSK-872 在 Z-VAD-fmk 存在的情况下阻断了 poly(I:C) 在成纤维细胞中诱导的 TLR3 诱导的坏死，两种病毒-和 TLR3 诱导的坏死独立于 Nec-1 对 RIP1 激酶的抑制，但对 GSK-872 的抑制敏感 [2,3,4]。药理学抑制剂 GSK-872 在体外和体内增强胰岛素信号，有助于改善胰岛素敏感性[9]。
当使用人类 HT-29 细胞在细胞培养物中进行评估时，GSK-872 结合激酶结构域并以高特异性抑制激酶活性，靶向更广泛的促坏死刺激物[1]。 RIP3i 化合物 GSK-872 以浓度依赖性方式阻断 TNF 诱导的坏死性凋亡。在基于细胞的测定中，与无细胞生化测定相比，IC50 有 100 到 1,000 倍的变化。 GSK-872 阻断从全血中分离的原代人嗜中性粒细胞的坏死性凋亡，并阻断小鼠细胞的坏死性凋亡。小鼠骨髓来源的巨噬细胞 (BMDM) 或巯基乙酸盐诱导的腹膜巨噬细胞 (PEC) 以及 3T3SA 成纤维细胞也受到 GSK-872 浓度的保护。
GSK-872 可显着减轻 SAH 大鼠的脑水肿并改善神经功能，并减少坏死细胞的数量。确定了 GSK-872 诱导 SAH 神经保护作用的确切机制[6,7]。GSK-872 抑制 RIPK3 可减轻 RIPK3 依赖性坏死性凋亡，减轻脑水肿，改善神经功能SAH。 GSK-872 还可以改善 CIH 暴露后喂食 HFCD 的小鼠的肝脂肪变性和肝损伤[8]。
| Cell experiment [1]: | |
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Cell lines |
HT-29 cells |
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Preparation Method |
Relative viability of human HT-29 cells 24 hr posttreatment (hpt) with TNF (10 ng/ml), zVAD-fmk (zVAD; 20 μM), and SMAC007 (100 nM) in the presence of increasing concentrations of GSK-872, assessed by determining ATP levels (mean ± range is shown) compared to cells treated with vehicle (DMSO) alone. |
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Reaction Conditions |
0.01, 0.03 , 0.1, 0.3, 1, and 3 μM；24 hours |
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Applications |
When evaluated in cell culture using human HT-29 cells,GSK-872 ( 0.01-3 μM; 24 hours) blocks TNF-induced necroptosis in human HT-29 cells in a concentration-dependent manne[1]. |
| Animal experiment [2]: | |
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Animal models |
Sprague-Dawley male rats with 300–320?g body weight |
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Preparation Method |
GSK-872 was diluted with 1% DMSO to a concentration of 25?mM, and 6?μL of GSK-872 or diluted DMSO was administrated by a syringe pump at 30?min after SAH as previously described, Neurological function (n = 24) was evaluated at 24 h and 72 h after operation. Brain edema (n = 6), western blot (n = 6), PI staining (n = 6) and HMGB1 immunofluorescence (n = 6) were evaluated at 72 h after SAH. |
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Dosage form |
6ul 25mM; 24?h and 72?h |
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Applications |
GSK-872 hydrochloride (25 mM; intracerebroventricular injection) can attenuate brain edema and improve neurological function following subarachnoid hemorrhage (SAH) and reduce the number of necrotic cells. GSK-872 hydrochloride can also decrease the protein levels of RIPK3 and MLKL, and cytoplasmic translocation and expression of HMGB1, an important pro-inflammatory protein[2]. |
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参考文献: [1]. Mandal P, Berger SB, et, al. RIP3 induces apoptosis independent of pronecrotic kinase activity. Mol Cell. 2014 Nov 20;56(4):481-95. doi: 10.1016/j.molcel.2014.10.021. Epub 2014 Nov 20. PMID: 25459880; PMCID: PMC4512186. [2]. Chen T, Pan H, et, al. Inhibiting of RIPK3 attenuates early brain injury following subarachnoid hemorrhage: Possibly through alleviating necroptosis. Biomed Pharmacother. 2018 Nov;107:563-570. doi: 10.1016/j.biopha.2018.08.056. Epub 2018 Aug 14. PMID: 30114640. |
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