| 名稱 | SKI II |
| 描述 | SKI II (SphK-I2) is a highly selective and non-ATP-competitive S1P receptor inhibitor (IC50: 0.5 μM) while exhibiting no inhibitory action on other kinases including PKCα, PI3K, and ERK2. |
| 細(xì)胞實(shí)驗(yàn) | T24, MCF-7, MCF-7/VP, and NCI/ADR cells are plated into 96-well tissue culture plates at ~15% confluency. After 24 hours, cells are treated with various concentrations of inhibitors. After an additional 48 hours, cell survival is assayed using the sulforhodamine B assay.(Only for Reference) |
| 激酶實(shí)驗(yàn) | SK Assay: A medium-throughput assay suitable for screening for inhibitors of recombinant human SK has been established. Briefly, 5 μg of purified GST-SK fusion protein are combined with 12 nM sphingosine, which contains a 100-fold dilution of [3-3H]sphingosine (20 Ci/mmol), 1 mM ATP, 1 mM magnesium chloride, and 200 μL of assay buffer [20 mM Tris HCl (pH 7.4), 20% glycerol, 1 mM beta-mercaptoethanol, 1 mM EDTA, 20 mM zinc chloride, 1 mM sodium orthovanadate, 15 mM sodium fluoride, and 0.5 mM 4-deoxypyridoxine]. Assays are run for 30 min at 25°C with shaking and contains either 1% DMSO or 5 g/mL test compound, which corresponds to concentrations of 10–25 μM. The reactions are terminated with 50 μL of concentrated ammonium hydroxide, followed by extraction of the assay mixture with chloroform:methanol (2:1). The aqueous portion is transferred to scintillation vials and radioactivity is quantified as a measure of [3H]]S1P formation using a Beckman LS 3801 Scintillation Counter. The intra-assay coefficient of variation is ~10%, whereas interassay variation is ~20%. |
| 體外活性 | SKI II（50 mg/kg,i.p.）通過(guò)抑制小鼠內(nèi)源性1-磷酸鞘氨醇生成,可改善抗原誘導(dǎo)的支氣管平滑肌高反應(yīng)性.在攜帶JC乳腺癌細(xì)胞的同源Balb/c小鼠實(shí)體瘤模型中, SKI II（50 mg/kg,i.p./p.o.）可顯著降低腫瘤生長(zhǎng),與對(duì)照組相比,無(wú)明顯的毒性或體重減輕. |
| 體內(nèi)活性 | 與降低S1P水平一致,SKI II可誘導(dǎo)T24細(xì)胞凋亡。在JC細(xì)胞中,SKI II可濃度依賴性降低S1P形成（IC50：12 μM）。在乳腺癌細(xì)胞系MDA-MB-231中,SKI II可明顯抑制內(nèi)源性SK活性。在人類癌細(xì)胞系,包括T-24,MCF-7,MCF-7/VP,NCI/ADR中,SKI II均有顯著的抗增殖效果（IC50：4.6/1.2/0.9/1.3 μM）。此外,通過(guò)下調(diào)P-gp的表達(dá),及通過(guò)下調(diào)SPHK1而上調(diào)凋亡,SKI II可逆轉(zhuǎn)SGC7901/DDP對(duì)順鉑的耐藥性。 |
| 存儲(chǔ)條件 | Powder: -20°C for 3 years | In solvent: -80°C for 1 year | Shipping with blue ice. |
| 溶解度 | DMSO : 40 mg/mL (132.11 mM), Sonication is recommended.
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| 關(guān)鍵字 | SKI II | Sphingosine kinase | Apoptosis | SphK | Wnt | Inhibitor | inhibit |
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