PF-543 hydrochloride

Catalog No.S7177 Batch:S717703

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Technical Data

Formula

C27H32ClNO4S
Molecular Weight 502.07 CAS No. 1706522-79-3
Solubility (25°C)* In vitro DMSO 100 mg/mL (199.17 mM)
Ethanol 100 mg/mL (199.17 mM)
Water Insoluble
In vivo (Add solvents to the product individually and in order)
Clear solution
5%DMSO 40%PEG300 5%Tween80 50%ddH2O
5.0mg/ml Taking the 1 mL working solution as an example, add 50 μL of 100 mg/ml clarified DMSO stock solution to 400 μL of PEG300, mix evenly to clarify it; add 50 μL of Tween80 to the above system, mix evenly to clarify; then continue to add 500 μL of ddH2O to adjust the volume to 1 mL. The mixed solution should be used immediately for optimal results. 
Clear solution
5% DMSO 95% Corn oil
0.8mg/ml Taking the 1 mL working solution as an example, add 50 μL of 16 mg/ml clear DMSO stock solution to 950 μL of corn oil and mix evenly. The mixed solution should be used immediately for optimal results. 
* <1 mg/ml means slightly soluble or insoluble.
* Please note that Selleck tests the solubility of all compounds in-house, and the actual solubility may differ slightly from published values. This is normal and is due to slight batch-to-batch variations.
* Room temperature shipping (Stability testing shows this product can be shipped without any cooling measures.)

Preparing Stock Solutions

Biological Activity

Description PF-543 hydrochloride, a novel sphingosine-competitive inhibitor of SphK1, inhibits SphK1 with IC50 and Ki of 2.0 nM and 3.6 nM, exhibits >100-fold selectivity over the SphK2 isoform. PF-543 hydrochloride induces apoptosis, necrosis, and autophagy.
Targets
SphK1 [1]
(Cell-free assay)		 SphK1 [1]
(Cell-free assay)
2.0 nM 3.6 nM(Ki)
In vitro

PF543 is a cell-permeable hydroxyl methylpyrrolidine compound that inhibits SphK-1/SphK1-catalyzed sphingosine phosphorylation in a reversible and sphingosine-competitive manner, exhibiting no affinity toward S1P receptors and much reduced inhibitory activity against Sphk2 (6.8% inhibition at 10 μM) or 46 other lipid and portein kinases (IC50 >10 μM). In the SphK1-overexpression 1483 head and neck carcinoma cells, PF-543 decreases the level of endogenous S1P 10-fold with a proportional increase in the level of sphingosine. PF-543 binds SphK1 reversibly (k off t1/2=8.5 min) and with high affinity and the binding constant (Kd) is 5 nM. PF543 had no effect on the proliferation and survival of 1483, A549, LN229, Jurkat, U937 and MCF-7 cells, despite a dramatic change in the cellular S1P/sphingosine ratio. PF-543 is effective as a potent inhibitor of S1P formation in whole blood, indicating that the SphK1 isoform of sphingosine kinase is the major source of S1P in human blood. [1]
In vivo

PF-543, a potent sphingosine kinase 1 inhibitor, reduced dysfunctional hypertrophy, associated with protection against cardiomyocyte apoptosis.
Features The most potent inhibitor of SphK1 described to date.

Protocol (from reference)

Kinase Assay:

[1]
  • FITC-S1P quantification/Caliper assay

    A 384-well format of the SphK enzyme assay based on separation of FITC-S1P from unreacted FITC-sphingosine substrate using a microfluidic capillary electrophoresis mobility-shift system is developed. Briefly, 3 nM SphK1–His6 is incubated with 1 μM FITC-sphingosine, 20 μM ATP and 10 μM compound (a final concentration of DMSO of 2 %) in a buffer containing 100 mM Hepes (pH 7.4), 1 mM MgCl2,0.01% Triton X-100, 10% glycerol, 100 μM sodium orthovanadate and 1 mM DTT for 1 h in a 384-well Matrical MP-101-1-PP plate. Reaction mixtures (10 μL) are quenched by the addition of 20 μL of 30 mM EDTA and 0.15% Coating Reagent-3 in 100 mM Hepes, and a small aliquot of each reaction (a few nanolitres) is analysed in the Caliper LabChip 3000 instrument under -1.5 psi (psi=6.9 kPa) pressure, a downstream voltage of -1900 V and a sip time of 0.2 s. Phosphorylated fluorescent product and unphosphorylated fluorescent substrate appeared as distinctive peaks and are quantified using the Caliper data.
Cell Assay:

[1]
  • Cell lines

    1483, A549, LN229, Jurkat, U937, MCF-7

  • Concentrations

    ~1 μM

  • Incubation Time

    7 days

  • Method

    CellTiter-Glo Assay
Animal Study:

[2]
  • Animal Models

    C57BL/6 mice

  • Dosages

    1 mg/kg

  • Administration

    i.p.

Customer Product Validation

Data from [Data independently produced by , , Int J Biochem Cell Biol, 2017, 90:17-28]

Data from [Data independently produced by , , Biochem Biophys Res Commun, 2016, 470(3):728-34.]

Selleck's PF-543 hydrochloride has been cited by 30 publications

Targeting the SphK1/S1P/PFKFB3 axis suppresses hepatocellular carcinoma progression by disrupting glycolytic energy supply that drives tumor angiogenesis [ J Transl Med, 2024, 22(1):43] PubMed: 38200582
Induction of Inflammation Disrupts the Negative Interplay between STING and S1P Axis That Is Observed during Physiological Conditions in the Lung [ Int J Mol Sci, 2023, 24(9)8303] PubMed: 37176007
Comprehensive metabolomics expands precision medicine for triple-negative breast cancer [ Cell Res, 2022, 10.1038/s41422-022-00614-0] PubMed: 35105939
Sphingosine kinase 1 promotes tumor immune evasion by regulating the MTA3-PD-L1 axis [ Cell Mol Immunol, 2022, 19(10):1153-1167] PubMed: 36050478
Sphingosine kinase 1 promotes tumor immune evasion by regulating the MTA3-PD-L1 axis [ Cell Mol Immunol, 2022, 19(10):1153-1167] PubMed: 36050478
Targeting sphingosine kinase 1/2 by a novel dual inhibitor SKI-349 suppresses non-small cell lung cancer cell growth [ Cell Death Dis, 2022, 13(7):602] PubMed: 35831279
The anti-osteosarcoma cell activity by the sphingosine kinase 1 inhibitor SKI-V [ Cell Death Discov, 2022, 8(1):48] PubMed: 35115496
The anti-osteosarcoma cell activity by the sphingosine kinase 1 inhibitor SKI-V [ Cell Death Discov, 2022, 8(1):48] PubMed: 35115496
S1P-Induced TNF-α and IL-6 Release from PBMCs Exacerbates Lung Cancer-Associated Inflammation [ Cells, 2022, 11-162524] PubMed: 36010601
Regulatory Role of Sphingosine-1-Phosphate and C16:0 Ceramide, in Immunogenic Cell Death of Colon Cancer Cells Induced by Bak/Bax-Activation [ Cancers (Basel), 2022, 14(21)5182] PubMed: 36358599

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SHIPPING AND STORAGE
Selleck products are transported at room temperature. If you receive the product at room temperature, please rest assured, the Selleck Quality Inspection Department has conducted experiments to verify that the normal temperature placement of one month will not affect the biological activity of powder products. After collecting, please store the product according to the requirements described in the datasheet. Most Selleck products are stable under the recommended conditions.

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