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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; Sphingosine kinase inhibitor II; SK-III) is a potent, highly selective, non-lipid and non-ATP-competitive sphingosine kinase (SphK) inhibitor with potential anticancer activity. It inhibits SphK with an IC50 of 0.5 μM, and displays no inhibitory effects against other closely related kinases such as 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; SKI II; Sphingosine kinase inhibitor II; SKIII

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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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