GSK-872
目录号 : GC19175
GSK-872 是一种 RIPK3 抑制剂。
Cas No.:1346546-69-7
Sample solution is provided at 25 µL, 10mM.
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Related Biological Data
Pharmacological blockade of autophagy neutralizes the effects of RIPK1/RIPK3 inhibition. (D, H) following treatment Nec-1 (100 μM) and GSK (10 μM). Representative images and percentages of LC3-II, P62, active-cathepsin D
Combination of Nec-1 (10μM) and GSK (1μM) (GLPBIO, USA) down-regulates the levels of RIPK1, ser166-p-RIPK1, RIPK3, ser232-p-RIPK3, alleviates cell death and restores autophagic flux of CFs.
Cell Death & Disease 13.2 (2022): 1-11. PMID: 35165268 IF: 8.469 -
Related Biological Data
GSK0 872 pretreatment inhibited necroptosis in cardiomyocytes. (A) RIPK3 was immunofluorescence stained by Alexa Fluor 488 (Green)-conjugated IgG. The nuclei were stained by DAPI (Blue).
In addition, cardiomyocytes were transfected with RIPK3 siRNA or RIPK3 inhibitor GSK 872 (10µM, GlpBio,Montclair, CA, USA) for pretreatment for 4h followed by AGEs stimulation for 24h.
J Exp Clin Canc Res 41.1 (2022): 1-17. PMID: 35805993 IF: 5.5999 -
Related Biological Data
SARS-CoV-2 E protein induced pyroptosis-like death in THP-1 cells. (A)The release of LDH to the supernatant and the cell viability were detected respectively using the commercial kits.
THP-1 cells were pretreated with different cell death inhibitors ZVAD (50μm), Fer-1 (3μm), Necro (3μm), and GSK-872 (GLPBIO, USA, 3μm) for 1h, followed by 1μg/mL of E protein stimulation for 4h.
Biochemistry and Cell Biology (2023). PMID: 36927169 IF: 2.8997
Quality Control & SDS
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- Purity: >99.50%
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- Datasheet
| 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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References: [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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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].
References:
[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]。
| Cas No. | 1346546-69-7 | SDF | |
| Canonical SMILES | O=S(C1=CC=C2N=CC=C(NC3=CC=C(SC=N4)C4=C3)C2=C1)(C(C)C)=O | ||
| 分子式 | C19H17N3O2S2 | 分子量 | 383.49 |
| 溶解度 | DMSO : ≥ 100 mg/mL (260.76 mM) | 储存条件 | Store at -20°C |
| General tips | 请根据产品在不同溶剂中的溶解度选择合适的溶剂配制储备液；一旦配成溶液，请分装保存，避免反复冻融造成的产品失效。 储备液的保存方式和期限：-80°C 储存时，请在 6 个月内使用，-20°C 储存时，请在 1 个月内使用。 为了提高溶解度，请将管子加热至37℃，然后在超声波浴中震荡一段时间。 |
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| Shipping Condition | 评估样品解决方案：配备蓝冰进行发货。所有其他可用尺寸：配备RT，或根据请求配备蓝冰。 | ||
| 制备储备液 | |||
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1 mg | 5 mg | 10 mg |
| 1 mM | 2.6076 mL | 13.0381 mL | 26.0763 mL |
| 5 mM | 0.5215 mL | 2.6076 mL | 5.2153 mL |
| 10 mM | 0.2608 mL | 1.3038 mL | 2.6076 mL |
| 第一步：请输入基本实验信息（考虑到实验过程中的损耗，建议多配一只动物的药量） | ||||||||||
| 给药剂量 | mg/kg |  |
动物平均体重 | g | |
每只动物给药体积 | ul | |
动物数量 | 只 |
| 第二步：请输入动物体内配方组成（配方适用于不溶于水的药物；不同批次药物配方比例不同，请联系GLPBIO为您提供正确的澄清溶液配方） | ||||||||||
| % DMSO % % Tween 80 % saline | ||||||||||
| 计算重置 | ||||||||||
计算结果:
工作液浓度： mg/ml；
DMSO母液配制方法： mg 药物溶于 μL DMSO溶液（母液浓度 mg/mL，
体内配方配制方法：取 μL DMSO母液，加入 μL PEG300，混匀澄清后加入μL Tween 80，混匀澄清后加入 μL saline，混匀澄清。
1. 首先保证母液是澄清的；
2.
一定要按照顺序依次将溶剂加入，进行下一步操作之前必须保证上一步操作得到的是澄清的溶液，可采用涡旋、超声或水浴加热等物理方法助溶。
3. 以上所有助溶剂都可在 GlpBio 网站选购。
Intermittent hypoxia aggravates non-alcoholic fatty liver disease via RIPK3-dependent necroptosis-modulated Nrf2/NFκB signaling pathway
Life Sci2021 Nov 15;285:119963.PMID: 34536498DOI: 10.1016/j.lfs.2021.119963
Aims: Hepatocyte necroptosis is a critical event in the progression of non-alcoholic fatty liver disease (NAFLD). Obstructive sleep apnea hypopnea syndrome (OSAHS) and chronic intermittent hypoxia (CIH) may be linked with the pathogenesis and the severity of NAFLD. However, the potential role of necroptosis in OSAHS-associated NAFLD has not been evaluated. The present study investigated whether IH could affect NAFLD progression through promoting receptor-interacting protein kinase-3 (RIPK3)-dependent necroptosis, oxidative stress, and inflammatory response, and further elucidated the underlying molecular mechanisms.
Main methods: LO2 cells were treated with palmitic acid (PA) and subjected to IH, and necroptosis, oxidative stress, and inflammation were assessed. The high-fat choline-deficient (HFCD)-fed mouse model was also used to assess the effects of CIH in experimental NAFLD in vivo.
Key findings: In this study, we found that RIPK3-mediated necroptosis was activated both in the PA plus IH-treated LO2 cells and liver of HFCD/CIH mice, and which could trigger oxidative stress and inflammatory response by decreasing Nrf2 and increasing p-P65. RIPK3 downregulation significantly reduced hepatocyte necroptosis, and ameliorated oxidative stress and inflammation through modulating Nrf2/NFκB pathway in vitro and vivo. Similarly, pretreatment with TBHQ, an activator of Nrf2, effectively blocked the generation of oxidative productions and inflammatory cytokines. In addition, RIPK3 inhibitor GSK-872 or TBHQ administration obviously alleviated hepatic injury, including histology, transaminase activities, and triglyceride contents in vivo.
Significance: IH aggravates NAFLD via RIPK3-dependent necroptosis-modulated Nrf2/NFκB signaling pathway, and which should be considered as a potential therapeutic strategy for the treatment of NAFLD with OSASH.
GDC-0326 Enhances the Effects of 5-Fu in Colorectal Cancer Cells by Inducing Necroptotic Death
Onco Targets Ther2021 Apr 13;14:2519-2530.PMID: 33880032DOI: 10.2147/OTT.S302334
Aim: Chemoresistance to 5-fluorouracil (5-Fu) is common in colorectal cancer (CRC). Programmed necrosis (necroptosis) is an alternative form of programmed cell death regulated by receptor-interacting protein kinase (RIPK) 1 and 3, assumed as a novel target of cancer therapy. In this study, we aimed to explore whether a novel small molecular agent GDC-0326 could facilitate the effect of 5-Fu through necroptosis.
Main methods: Cell Counting Kit-8 (CCK-8) assay and colony formation were performed to confirm the function of GDC-0326 in CRC cells. Western blot and immunofluorescence were conducted to measure the altered expressions of RIPK1/RIPK3 induced by GDC-0326. Subcutaneous tumor models were used to evaluate the chemotherapeutic effects and concomitant side effects of GDC-0326 in vivo.
Key findings: We found that GDC-0326 effectively suppressed the growth of CRC cells in a dose-dependent manner. The induction of necroptosis by GDC-0326 was correlated with the modulation of RIPK1 and RIPK3. Necrostatin-1 and GSK-872, inhibitors of RIPK1 and RIPK3, respectively, could rescue the cell death induced by GDC-0326. In addition, in vitro and in vivo studies showed that 5-Fu plus GDC-0326 evinced a better antitumor efficacy by suppressing tumor growth and increasing tumor necrosis with no increased toxicity.
Significance: This study demonstrates that GDC-0326 plus 5-Fu has augmented antitumor efficacy and acceptable safety, which might be a promising therapeutic strategy for CRC patients in the future.
Inhibition of RIPK1/RIPK3 ameliorates osteoclastogenesis through regulating NLRP3-dependent NF-κB and MAPKs signaling pathways
Biochem Biophys Res Commun2020 Jun 11;526(4):1028-1035.PMID: 32321638DOI: 10.1016/j.bbrc.2020.03.177
Osteoblast-induced bone formation and osteoclast-regulated bone resorption are the essential events contributing to bone homeostasis. It is critical to investigate the underlying molecular mechanisms. In this study, we explored the effects of receptor-interacting serine-threonine kinases (RIPKs) on osteoclastogenesis and bone loss in vitro and in vivo. We found that both RIPK1 and RIPK3 expression levels were highly up-regulated during osteoclastogenesis. Inhibiting RIPK1 and RIPK3 by their inhibitors Necrostatin-1 (Nec-1) and GSK-872, respectively, showed effective activities against osteoclast differentiation and bone resorption induced by receptor activator of nuclear factor-κB ligand (Rankl). Osteoclast-specific gene expression levels were also impeded by RIPK1/RIPK3 blockage in a time-dependent manner. Subsequently, we found that the pyrin domain-containing protein 3 (NLRP3) inflammasome stimulated by Rankl during osteoclastogenesis was greatly inhibited by Nec-1 and GSK-872. Additionally, reducing RIPK1/RIPK3 overtly reduced the activation of NF-κB (p65) and mitogen-activated protein kinases (MAPKs) signaling during Rankl-induced osteoclast formation. Notably, adenovirus-regulated NLRP3 over-expression significantly abrogated the inhibitory effects of Nec-1 and GSK-872 on NF-κB and MAPKs signaling pathways, as well as the osteoclastogenesis. Finally, the in vivo studies indicated that suppressing RIPK1/RIPK3 could effectively ameliorate ovariectomy (OVX)-induced bone loss in mice through repressing osteoclastogenesis, as proved by the clearly down-regulated number of osteoclasts via histological staining. In conclusion, our study elucidated that restraining RIPK1/RIPK3 could hinder osteoclastogenesis and attenuate bone loss through suppressing NLRP3-dependent NF-κB and MAPKs signaling pathways. Therefore, targeting RIPK1/RIPK3 signaling might be a potential therapeutic strategy to develop effective treatments against osteoclast-related bone lytic diseases.