Background
SB 216763 stands out as a powerful, selective, and ATP-competitive inhibitor of GSK-3, effectively targeting both GSK-3α and GSK-3β with an impressive IC50 of 34.3 nM[1-3].
SB 216763(1-20 μM; 24 hours) induces β-catenin mediated-transcription in a dose-dependent manner. Besides, SB 216763(10, 15 and 20 μM) can maintain mouse embryonic stem cells (mESCs) in a pluripotent state in the absence of exogenous leukemia inhibitory factor (LIF) when cultured on mouse embryonic fibroblasts (MEFs) [4]. SB 216763 enhances neurogenesis but does not alter cell cycle exit or cell survival[5].
SB 216763(1-1.5 mg/kg/day；4weeks；s.c) was able to reverse the increased expression levels of molecular markers of inflammation and fibrosis in the heart and kidneys induced by aldosterone (Aldo) injection. Additionally, SB 216763 effectively suppressed cardiac and renal inflammatory cytokine levels, including TNF-a, IL-1β, and MCP-1[6]. SB 216763(10 mg/kg；i.p;3weeks) can reduce delayed gastric emptying (DGE) in high-fat diet (HFD)-induced obese/Type 2 diabetes (T2D) female mice[7]. Pretreatment with SB 216763 (2.5-5 mg/kg, i.p.) prior to amphetamine (2 mg/kg, i.p.) significantly reduced amphetamineinduced ambulation and stereotypy[8].
SB 216763是一种强效、选择性且ATP竞争性的GSK-3抑制剂，显著作用于GSK-3α和GSK-3β，其IC50仅为34.3 nM[1-3]。
SB 216763(1-20μM;24h) 呈剂量依赖性诱导β-catenin介导的转录。此外，SB 216763(10、15和20μM)在小鼠胚胎成纤维细胞(MEF)培养时，可以在缺乏外源性白血病抑制因子(LIF)的情况下维持小鼠胚胎干细胞(mESCs)的多能状态[4]。SB 216763促进神经发生，但不改变细胞周期退出或细胞存活[5]。
SB 216763（1-1.5 mg/kg/day；4weeks；s.c）能够逆转醛固酮(Aldo)注射引起的心脏和肾脏炎症和纤维化分子标志物的表达水平升高。此外，SB 216763(10 mg/kg；i.p;3weeks)有效抑制心脏和肾脏炎症细胞因子水平，包括TNF-a、IL-1β和MCP-1[6]。SB 216763可降低高脂饮食(HFD)诱导的肥胖/ 2型糖尿病(T2D)雌性小鼠胃排空延迟(DGE) [7]。在使用安非他明之前使用SB 216763 (2.5-5 mg/kg, i.p.)进行预处理可显著降低安非他明引起的活动和刻板行为[8]。
参考文献:
[1]. Wang M, Gao M, et,al. The first synthesis of [(11)C]SB-216763, a new potential PET agent for imaging of glycogen synthase kinase-3 (GSK-3). Bioorg Med Chem Lett. 2011 Jan 1;21(1):245-9. doi: 10.1016/j.bmcl.2010.11.026. Epub 2010 Nov 11. PMID: 21115250.
[2]. Coghlan MP, Culbert AA, et,al. Selective small molecule inhibitors of glycogen synthase kinase-3 modulate glycogen metabolism and gene transcription. Chem Biol. 2000 Oct;7(10):793-803. doi: 10.1016/s1074-5521(00)00025-9. PMID: 11033082.
[3]. Xu Y, Zhao J, et,al.Derivation of totipotent-like stem cells with blastocyst-like structure forming potential. Cell Res. 2022 Jun;32(6):513-529. doi: 10.1038/s41422-022-00668-0. Epub 2022 May 4. PMID: 35508506; PMCID: PMC9160264.
[4]. Kirby LA, Schott JT, et,al.Glycogen synthase kinase 3 (GSK3) inhibitor, SB-216763, promotes pluripotency in mouse embryonic stem cells. PLoS One. 2012;7(6):e39329. doi: 10.1371/journal.pone.0039329. Epub 2012 Jun 26. PMID: 22745733; PMCID: PMC3383737.
[5]. Lange C, Mix E, et,al. Small molecule GSK-3 inhibitors increase neurogenesis of human neural progenitor cells. Neurosci Lett. 2011 Jan 13;488(1):36-40. doi: 10.1016/j.neulet.2010.10.076. Epub 2010 Nov 5. PMID: 21056624.
[6]. Zhang YD, Ding XJ, et,al. SB-216763, a GSK-3β inhibitor, protects against aldosterone-induced cardiac, and renal injury by activating autophagy. J Cell Biochem. 2018 Jul;119(7):5934-5943. doi: 10.1002/jcb.26788. Epub 2018 Mar 30. PMID: 29600538; PMCID: PMC6001754.
[7]. Sampath C, Srinivasan S, et,al.Inhibition of GSK-3β restores delayed gastric emptying in obesity-induced diabetic female mice. Am J Physiol Gastrointest Liver Physiol. 2020 Oct 1;319(4):G481-G493. doi: 10.1152/ajpgi.00227.2020. Epub 2020 Aug 19. PMID: 32812777; PMCID: PMC7654647.
[8]. Enman NM, Unterwald EM. Inhibition of GSK3 attenuates amphetamine-induced hyperactivity and sensitization in the mouse. Behav Brain Res. 2012 May 16;231(1):217-25. doi: 10.1016/j.bbr.2012.03.027. PMID: 22649795; PMCID: PMC3566781.
Protocol
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Enzyme inhibition assay[1]:
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Preparation method
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GSK-3 kinase activity was measured, in the presence or absence of SB 216763, in a reaction mixture containing final concentrations of: 1 nM human GSK-3α or rabbit GSK3α; 50 mM MOPS pH 7.0; 0.2 mM EDTA; 10 mM Mg-acetate; 7.5 mM β-mercaptoethanol; 5% (w/v) glycerol; 0.01% (w/v) Tween-20; 10% (v/v) DMSO; 28 μM GS-2 peptide substrate.
The GS-2 peptide sequence corresponds to a region of glycogen synthase that is phosphorylated by GSK-3 as previously described.
The assay was initiated by the addition of 0.34 μCi [33P]γ-ATP (IC50 determinations) or 2.7 μCi [33P]γ-ATP (Ki determinations).
The total ATP concentration was 10 μM (IC50 determinations) or ranged from 0 to 45 μM (Ki determinations). Following 30 min incubation at room temperature the assay was stopped by the addition of one third assay volume of 2.5% (v/v) H3PO4 containing 21 mM ATP.
Samples were spotted onto P30 phosphocellulose mats and these were washed six times in 0.5% (v/v) H3PO4.
The filter mats were sealed into sample bags containing Wallac betaplate scintillation fluid. 33P incorporation into the substrate peptide was determined by counting the mats in a Wallac microbeta scintillation counter.
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Applications
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SB 216763 inhibited human GSK-3α with IC50 of 34 nM, when assayed in the presence of 0.01 mM ATP.
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Cell experiment[2]:
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Cell lines
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HEK293 cells
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Preparation method
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Cells were electroporated with the Super TOPFlash and pGL4.73 Renilla luciferase reporters. The cells were treated in quadruplicate wells with a vehicle control (0.1% DMSO) or the indicated doses of SB 216763 for 24 hours.
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Reaction Conditions
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1–20 μM SB 216763；24 hours
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Applications
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SB 216763 induces β-catenin mediated-transcription in a dose-dependent manner.
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Animal experiment[3]:
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Animal models
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Adult male Wistar rats
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Preparation method
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The animals were divided into four groups (n = 9/group): (1) Vehicle infusion group treated with vehicle alone; (2) Aldo-salt group treated with an infusion of Aldo-salt (1 mg/kg/day diluted in sunflower oil and administered by subcutaneous injection); (3) Aldo-salt plus SB 216763 group treated with an infusion of Aldo-salt plus SB 216763 at 1.5 mg/kg/day; and (4) SB 216763 group.
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Dosage form
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1-1.5 mg/kg/day；4weeks；s.c
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Applications
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SB 216763 suppresses cardiac inflammation and fibrosis caused by Aldo-salt.
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参考文献:
[1]. ?Coghlan MP, Culbert AA, et,al. Selective small molecule inhibitors of glycogen synthase kinase-3 modulate glycogen metabolism and gene transcription. Chem Biol. 2000 Oct;7(10):793-803. doi: 10.1016/s1074-5521(00)00025-9. PMID: 11033082.
[2]. Kirby LA, Schott JT, et,al. Glycogen synthase kinase 3 (GSK3) inhibitor, SB-216763, promotes pluripotency in mouse embryonic stem cells. PLoS One. 2012;7(6):e39329. doi: 10.1371/journal.pone.0039329. Epub 2012 Jun 26. PMID: 22745733; PMCID: PMC3383737.
[3].Zhang YD, Ding XJ, et,al.SB-216763, a GSK-3β inhibitor, protects against aldosterone-induced cardiac, and renal injury by activating autophagy. J Cell Biochem. 2018 Jul;119(7):5934-5943. doi: 10.1002/jcb.26788. Epub 2018 Mar 30. PMID: 29600538; PMCID: PMC6001754.
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