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    SKI II
    SKI II

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    This product is for research use only, not for human use. We do not sell to patients.
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    InvivoChem Cat #: V1503
    CAS #: 312636-16-1Purity ≥98%

    Description: SKI II (also known as SphK-I2) is a potent, highly selective, non-lipid and non ATP-competitive sphingosine kinase (SphK) inhibitor with IC50 of 0.5 μM, while it exhibits no inhibitory effects on other kinases including PI3K, PKCα and ERK2. SKI-II inhibits acute myelogenous leukemia cell growth in vitro and in vivo. SKI-II was more efficient than two known SphK1 inhibitors SK1-I and FTY720 in inhibiting AML cells. SKI-II treatment inhibited SphK1 activation, and concomitantly increased level of sphingosine-1-phosphate (S1P) precursor ceramide in AML cells. 

    References: Cancer Res. 2003 Sep 15;63(18):5962-9; J Pharmacol Exp Ther. 2006 Aug;318(2):596-603.

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    Molecular Weight (MW)302.78
    CAS No.312636-16-1
    Storage-20℃ for 3 years in powder form
    -80℃ for 2 years in solvent
    Solubility (In vitro)DMSO: 61 mg/mL (201.5 mM)
    Water: <1 mg/mL
    Ethanol: 61 mg/mL (201.5 mM)
    Solubility (In vivo)30% PEG400+0.5% Tween80+5% Propylene glycol: 20 mg/mL 
    SynonymsSphK-I2; SKI-II;  SKIII; SKI II. Sphingosine kinase inhibitor II

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    In Vitro

    In vitro activity: SKI II potently inhibits endogenous SK activities in the breast cancer cell line MDA-MB-231. SKI II demonstrates significantly antiproliferative effects in human cancer cell lines including T-24, MCF-7, MCF-7/VP, NCI/ADR with IC50 of 4.6 μM , 1.2 μM, 0.9 μM and 1.3 μM, respectively. SKI II also induces apoptosis of T24 cells consistent with the hypothesized consequence of reducing S1P levels. As demonstrated previously in MDA-MB-231 cells, SKI II decreases S1P formation in JC cells in a concentration-dependent manner, with IC50 of 12 μM. In addition, SKI-II can reverse drug resistance of SGC7901/DDP to cisplatin by down-regulating expression of P-gp and up-regulating apoptosis through down-regulation of SphK1.

    Kinase 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%. 

    Cell Assay: 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.

    In VivoSKI II (50 mg/kg) after intraperitoneal or oral administration significantly decreases tumor growth a syngeneic Balb/c mouse solid tumor model that uses JC mammary adenocarcinoma cells relative to control groups with no overt toxicity or weight loss. SKI-II (50 mg/kg ip) ameliorates antigen-induced bronchial smooth muscle hyperresponsiveness in mice by endogenously inhibiting generation of S1P.
    Animal modelJC xenografts are established in mice
    Formulation & DosageDissolved in DMSO (ip), polyethylene glycol 400 (oral);  100 mg/kg; i.p. injection or oral administration

    Cancer Res. 2003 Sep 15;63(18):5962-9; J Pharmacol Exp Ther. 2006 Aug;318(2):596-603.

    These protocols are for reference only. InvivoChem does not independently validate these methods.


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