Background
PF-4708671, is a novel cell-permeable inhibitor of S6K1, specifically inhibits the S6K1 isoform with a Ki of 20 nM and IC50 of 160 nM. [1].
PF-4708671 prevents the S6K1-mediated phosphorylation of S6 protein in response to IGF-1 (insulin-like growth factor 1), PF-4708671 was also found to induce phosphorylation of the T-loop and hydrophobic motif of S6K1, an effect that is dependent upon mTORC1 (mTOR complex 1) [1]. RSK1, RSK2 and MSK1 were the other kinases inhibited by PF-4708671 (IC50 = 4.7 μM, 9.2 μM and 0.95 μM, respectively). PF-4708671 decreased phosphorylation of ribosomal protein S6 in HEK-293 cells. PF-4708671 has been used as the standard S6K1 inhibitor for the investigation of the role of S6K1 in several cancers. PF-4708671 in combination with tamoxifen was shown to be highly effective against ER+ MCF7 cells that had overexpression of S6K1 [2] and enhanced cell death in glucose-starved MCF7 cells via downregulation of anti-apoptotic proteins Mcl-1 and survivin [3].
PF-4708671 inhibite the AKT/mTOR/S6K1 pathway led to the inhibition of cell migration in triple-negative MDA-MB-231 cells and inhibition of local relapse in mice models [4]. Moreover, PF-4708671 improved glucose tolerance in high-fat -fed obese mice by restoring Akt S473 phosphorylation in metabolic tissues [5]. PF-4708671 has protective effects against NMDA-induced retinal neurotoxicity in rats [6].
参考文献:
[1]. Pearce, L. R. et al. Characterization of PF-4708671, a novel and highly specific inhibitor of p70 ribosomal S6 kinase (S6K1). Biochem. J. 431, 245-255 (2010).
[2]. Hong SE, Kim EK, Jin HO, Kim HA, Lee JK, Koh JS, et al. S6K1 inhibition enhances tamoxifen-induced cell death in MCF-7 cells through translational inhibition of Mcl-1 and survivin. Cell Biol Toxicol. 2013;29(4):273-82.
[3]. Choi HN, Jin HO, Kim JH, Hong SE, Kim HA, Kim EK, et al. Inhibition of S6K1 enhances glucose deprivation-induced cell death via downregulation of anti-apoptotic proteins in MCF-7 breast cancer cells. Biochem Biophys Res Commun. 2013;432(1):123-8.
[4]. Segatto I, Berton S, Sonego M, Massarut S, D'Andrea S, Perin T, et al. Inhibition of breast cancer local relapse by targeting p70S6 kinase activity. J Mol Cell Biol. 2013;5(6):428-31.
[5]. M. Shum, K. Bellmann, P. St-Pierre, A. Marette. Pharmacological inhibition of S6K1 increases glucose metabolism and Akt signalling in vitro and in diet-induced obese mice .Diabetologia, 59 (2016), pp. 592-603
[6]. Hayashi, I.; Aoki, Y.; Ushikubo, H.; Asano, D.; Mori, A.; Sakamoto, K.; Nakahara, T.; Ishii, K. Protective effects of PF-4708671 againstN-methyl-d-aspartic acid-induced retinal damage in rats. Fundam. Clin. Pharmacol. 2016, 30, 529-536.
PF-4708671 是一种新型的细胞渗透性 S6K1 抑制剂，特异性抑制 S6K1 亚型，Ki 为 20 nM，IC50 为 160 nM。 [1].
PF-4708671 阻止 S6K1 介导的 S6 蛋白磷酸化以响应 IGF-1（胰岛素样生长因子 1），PF-4708671 还被发现诱导 S6K1 的 T 环和疏水基序磷酸化，一种依赖于 mTORC1（mTOR 复合体 1）[1] 的效应。 RSK1、RSK2 和 MSK1 是其他被 PF-4708671 抑制的激酶（IC50 分别为 4.7 μM、9.2 μM 和 0.95 μM）。 PF-4708671 降低 HEK-293 细胞中核糖体蛋白 S6 的磷酸化。 PF-4708671 已被用作标准 S6K1 抑制剂，用于研究 S6K1 在多种癌症中的作用。 PF-4708671 与他莫昔芬联用被证明对过度表达 S6K1 [2] 的 ER+ MCF7 细胞非常有效，并通过下调抗凋亡蛋白 Mcl 增强葡萄糖饥饿 MCF7 细胞的细胞死亡-1 和生存 [3]。
PF-4708671 抑制 AKT/mTOR/S6K1 通路导致抑制三阴性 MDA-MB-231 细胞中的细胞迁移并抑制小鼠模型的局部复发[4]。此外，PF-4708671 通过恢复代谢组织中的 Akt S473 磷酸化，改善了高脂喂养肥胖小鼠的葡萄糖耐量[5]。 PF-4708671对NMDA诱导的大鼠视网膜神经毒性具有保护作用[6]。
Protocol
| Cell experiment [1]: |
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Cell lines
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A549 (adenocarcinoma), NCI-H460 (large cell carcinoma), and SK-MES-1 (squamous cell carcinoma) cells
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Preparation Method
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Tumor cells were seeded in 96-well plates at a density of 5×103 per well. After incubation in presence of PF-4708671 (0.1μM, 0.3μM, 1μM, 3μM and 10μM) for 24, 48 and 72 hours, respectively,
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Reaction Conditions
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0.1μM, 0.3μM, 1μM, 3μM and 10μM for 24, 48 and 72 hours
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Applications
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Proliferation abilities of the three NSCLC cell lines were significantly inhibited by PF-4708671. After 24 hours of treatment, PF-4708671 inhibited H460 cell proliferation at 10μM, and A549 and SK-MES-1 cell amounts were significantly reduced at 3μM and 0.1μM, respectively. In addition, H460, A549, and SK-MES-1 cell growth rates were significantly inhibited by PF-4708671 at 0.3μM, 0.1μM, and 0.1μM, respectively, 48 hours post treatment. All cell lines showed significantly reduced proliferation at 72 hour after treatment with 0.1μM PF-4708671.
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| Animal experiment [2]: |
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Animal models
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Male C57Bl/6 mice (6 weeks old)
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Preparation Method
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Mice were randomly assigned to three groups: (1) control (HF) receiving vehicle (8% EtOH [vol./vol.], 0.2% [wt/vol.] carboxymethylcellulose sterile); (2) treated with PF-4708671 (35 mg kg-1 day-1, i.p.); or (3) treated with rapamycin (2 mg kg-1 day-1, i.p.) for 7 days while being kept on the same high-fat diet.
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Dosage form
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Intraperitoneal injection, 35 mg kg-1 day-1 for 7 days.
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Applications
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PF-4708671 did not affect body weight or adiposity, 1 week of PF-4708671 treatment was found to improve fasting glucose whereas rapamycin further increased fasting hyperglycaemia in obese mice.
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参考文献:
[1]: Qiu ZX, Sun RF, Mo XM and Li WM: The p70S6K specific inhibitor PF-4708671 impedes non-small cell lung cancer growth. PLoS One. 11:e01471852016.
[2]: M. Shum, K. Bellmann, P. St-Pierre, A. Marette. Pharmacological inhibition of S6K1 increases glucose metabolism and Akt signalling in vitro and in diet-induced obese mice .Diabetologia, 59 (2016), pp. 592-603
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