PF-543

Catalog No.S7177

For research use only.

PF-543, 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 induces apoptosis, necrosis, and autophagy.

PF-543 Chemical Structure

CAS No. 1706522-79-3

Selleck's PF-543 has been cited by 17 publications

Purity & Quality Control

Choose Selective SPHK Inhibitors

Biological Activity

Description PF-543, 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 induces apoptosis, necrosis, and autophagy.
Features The most potent inhibitor of SphK1 described to date.
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]

Cell Data
Cell Lines Assay Type Concentration Incubation Time Formulation Activity Description PMID
human PAMSC cells MWnGeY5kfGmxbjDhd5NigQ>? MWCxNFAhdk1? MWeyOEBp NHjvW|RF\WO{ZXHz[UBw\iCVS{Gg[ZhxemW|c3nvckBqdiCqdX3hckBRSU2VQzDj[YxteyCjdDCxNFAhdk1iYX\0[ZIhOjRiaILzJIJ6KFenc4Tldo4h[myxdDDhcoFtgXOrcx?= NH24U5Q9[SC2YYLn[ZQ:L1:kbHHub{chcHKnZk2nbJR1eHN8Lz;weYJu\WRwbnPibU5vdG1wbnnoModwfi9{NEO5OlU4OCd-MkSzPVY2PzB:L3G+
Sf9 cells MWPGeY5kfGmxbjDhd5NigQ>? MWHJcohq[mm2aX;uJI9nKGi3bXHuJJJm[2:vYnnuZY51KFOyaFuxJIV5eHKnc4Pl[EBqdiCVZkmgZ4VtdHNiYYPz[ZN{\WRiYYOgdoFlcW:uYXLlcIVlKHC{b3T1Z5R{KHW|aX7nJFUhfU1ic4DobY5od3OrbnWgZY5lKDFyIIXNJIdidW2jW{OyVH1CXFBiYomgcIlyfWmmIIPjbY51cWyuYYTpc44h[2:3boTpcoc> M2TiW|xiKHSjcnfleF0oZ2KuYX7rK{BpemWoPTfoeJRxezpxL4D1Zo1m\C6wY3LpMo5tdS6waXiu[493NzJ3NkSzNFc1Lz5{NU[0N|A4PDxxYU6=
Sf21 cells MWrGeY5kfGmxbjDhd5NigQ>? M3rQeFkxKG2rboO= MkjDTY5pcWKrdHnvckBw\iCKaYO2JJRi\2enZDDoeY1idiCVcHjLNUBmgHC{ZYPz[YQhcW5iU3[yNUBk\WyuczD1d4lv\yCISWTDMZNxcGmwZ3;zbY5mKGG|IIP1ZpN1emG2ZTDwdoVqdmO3YnH0[YQh\m:{IEmwJI1qdnNicILpc5Ihe3Wkc4TyZZRmKGGmZHn0bY9vKGK7IFPhcIlx\XJiYYPzZZktKEurPUSuN{BvVQ>? M3HEblxiKHSjcnfleF0oZ2KuYX7rK{BpemWoPTfoeJRxezpxL4D1Zo1m\C6wY3LpMo5tdS6waXiu[493NzJ3NUG2O|k{Lz5{NUWxOlc6OzxxYU6=
PC3 NEfzWVlCdnSrcILvcIln\XKjdHn2[UBie3OjeR?= MYW3NkBpenN? NUjlboU3SW62aYDyc4xq\mW{YYTpeoUh[WO2aY\peJkh[WejaX7zeEBpfW2jbjDQR|Mh[2WubIOgZYZ1\XJiN{KgbJJ{KGK7IF3UV{Bie3OjeTygS2k2OD1zOd88UU4> Mn3EQIEhfGG{Z3X0QUdg[myjbnunJIhz\WZ;J3j0eJB{Qi9xcIXicYVlNm6lYnmucoxuNm6raD7nc5YwOjZ5OECzNFQoRjJ4N{iwN|A1RC:jPh?=
HEK293 MVnGeY5kfGmxbjDhd5NigQ>? MWCxNEB2VQ>? MX6xOkBpenN? NEnkc2RKdmS3Y4Tpc44hd2ZiU1uxJIRm\3KjZHH0bY9vKGmwIFjFT|I6OyClZXzsd{BifCBzMDD1UUBi\nSncjCxOkBpenNiYomge4V{fGW{bjDicI91fGmwZzDhcoFtgXOrczDpckBxemW|ZX7j[UBw\iCycn;0[YF{d22nIHnubIljcXSxcjDNS|E{Og>? NHSwcpI9[SC2YYLn[ZQ:L1:kbHHub{chcHKnZk2nbJR1eHN8Lz;weYJu\WRwbnPibU5vdG1wbnnoModwfi9{Nke4NFMxPCd-Mk[3PFA{ODR:L3G+
MDA1483 MkXnSpVv[3Srb36gZZN{[Xl? NF7hTYw{OCCvaX7z MYPJcohq[mm2aX;uJI9nKFOSSFuxJIlvKGi3bXHuJG1FSTF2OEOgZ4VtdHNidYPpcochSzF5LYPwbIlv\2:|aX7lJIF{KHO3YoP0doF1\SCjc4Pld5Nm\CCjczDy[YR2[3Srb36gbY4hSzF5LWOxVEBnd3KvYYTpc44heHKnaX7jeYJifGWmIH\vdkA{OCCvaX7zJJdqfGhic4Xid5Rz[XSnIH3lZZN2emWmIHHmeIVzKDF3IH3pcpMh[nliTFOtUXMh[W6jbInzbZMtKEmFNUC9NE4xODB6zszNMi=> MWW8ZUB1[XKpZYS9K39jdGGwazegbJJm\j1paIT0dJM7Ny:ydXLt[YQvdmOkaT7ucI0vdmmqLnfvek8zQDJ|MUSzN{c,Ojh{M{G0N|M9N2F-
Sf9 MoDoSpVv[3Srb36gZZN{[Xl? M{fSTlE2KG2rboO= M4K2OWlvcGmkaYTpc44hd2ZiaIXtZY4hSy22ZYLtbY5idCCKaYO2MZRi\2enZDDTVGhMOSCneIDy[ZN{\WRiaX6g[oFtdCCjcn35e49zdSC|ZkmgZ4VtdHNiYYPz[ZN{\WRiYYOgdoVlfWO2aX;uJIlvKEGGUDDmc5Ju[XSrb36geZNqdmdic4DobY5od3OrbnWgZZMhe3Wkc4TyZZRmKHC{ZXnuZ5Vj[XSnZDDmc5IhOTVibXnud{Bnd2yub4fl[EBjgSC|dXLzeJJifGViYXTkbZRqd25iYX\0[ZIhOSCqcjDifUB1emGwc3Py[YVv\XJvYnHz[YQh\my3b4Lld4NmdixiSVO1NF0xNjByMt88UU4> MnH3QIEhfGG{Z3X0QUdg[myjbnunJIhz\WZ;J3j0eJB{Qi9xcIXicYVlNm6lYnmucoxuNm6raD7nc5YwOjh{M{G0N|MoRjJ6MkOxOFM{RC:jPh?=
Sf9 MXXGeY5kfGmxbjDhd5NigQ>? M4rYeVEhcHJ? NEDDTGFKdmirYnn0bY9vKG:oIHj1cYFvKENvdHXycYlv[WxiSHnzOk11[WepZXSgV3BJUzFiZYjwdoV{e2WmIHnuJIZidGxiYYLtfZdwem1ic3[5JINmdGy|IHHzd4V{e2WmIHHzJJJm\HWldHnvckBqdiCVMWCg[o9zdWG2aX;uJJV{cW6pIIPwbIlv\2:|aX7lJIF{KHO3YoP0doF1\SCjZoTldkAyKGi{IHL5JGZKXENvYnHz[YQh[2GuaYDldkBie3OjeTygTWM2OD1yLkCwNlfPxE1w M1XR[VxiKHSjcnfleF0oZ2KuYX7rK{BpemWoPTfoeJRxezpxL4D1Zo1m\C6wY3LpMo5tdS6waXiu[493NzJ6MkOxOFM{Lz5{OEKzNVQ{OzxxYU6=
Sf21 MnKxSpVv[3Srb36gZZN{[Xl? NYLVN|NkOSCqch?= Mk[4TY5pcWKrdHnvckBw\iC{ZXPvcYJqdmGwdDDoeY1idiCFLYTldo1qdmGuIFjpd{11[WepZXSgV3BJUzFiZYjwdoV{e2WmIHnuJIZidGxiYYLtfZdwem1ic3[yNUBk\WyuczD1d4lv\yC|cHjpcodwe2mwZTDhd{B{fWK|dILheIUh[W[2ZYKgNUBpeiCkeTDGTXRENWKjc3XkJINidGmyZYKgZZN{[XluIFvpQVAvODB|Nt88UU4> NV24bVFLRGFidHHy[4V1RSehYnzhcosoKGi{ZX[9K4h1fHC|Oj:vdJVjdWWmLn7jZokvdmyvLn7pbE5od3ZxMki0NFY3PDZpPkK4OFA3PjR4PD;hQi=>
Sf21 NFS4VJFHfW6ldHnvckBie3OjeR?= NXjoVVNGOSCqch?= NHnVN|FKdmirYnn0bY9vKG:oIILlZ49u[mmwYX70JGMufGW{bXnuZYwhUGm|Nj30ZYdo\WRiaIXtZY4hW3CqS{Gg[ZhxemW|c3XkJIlvKGKjY4Xsc5ZqenW|IHnu[oVkfGWmIIPmNlEh[2WubIOgeZNqdmdiRlnUR{1{eGirbnfvd4lv\SCjczDzeYJ{fHKjdHWgZYZ1\XJiMTDodkBqdiCycnXz[Y5k\SCxZjDBWHAh[nlibXnjdo9ndHWrZHnjJINieGmubHHyfUBmdGWldILvdIhwemW|aYOgcY9jcWyrdImgd4hq\nRiYYPzZZktKEmFNUC9NE4xODJ5zszNMi=> M2Hh[FxiKHSjcnfleF0oZ2KuYX7rK{BpemWoPTfoeJRxezpxL4D1Zo1m\C6wY3LpMo5tdS6waXiu[493NzJ6NEC4NVkxLz5{OESwPFE6ODxxYU6=
Sf21 M{fjO2Z2dmO2aX;uJIF{e2G7 M2DhUVEhcHJ? MmfsTY5pcWKrdHnvckBw\iCqdX3hckBz\WOxbXLpcoFvfCCFLYTldo1qdmGuIFjpd|YhfGGpZ3XkJHNxcEt{IHX4dJJme3OnZDDpckBj[WO3bH;2bZJ2eyCrbn\lZ5Rm\CC|ZkKxJINmdGy|IIXzbY5oKE[LVFOtd5BpcW6pb4PpcoUh[XNic4Xid5Rz[XSnIHHmeIVzKDFiaIKgbY4heHKnc3XuZ4Uhd2ZiQWTQJIJ6KG2rY4Lv[ox2cWSrYzDjZZBqdGyjcomg[Yxm[3S{b4Doc5Jme2m|IH3vZoltcXS7IIPobYZ1KGG|c3H5MEBKSzVyPUCuN|bPxE1w NEXSfIc9[SC2YYLn[ZQ:L1:kbHHub{chcHKnZk2nbJR1eHN8Lz;weYJu\WRwbnPibU5vdG1wbnnoModwfi9{OESwPFE6OCd-Mki0NFgyQTB:L3G+
Sf21 NGfHXIdHfW6ldHnvckBie3OjeR?= MlnOTY5pcWKrdHnvckBw\iCqdX3hckBENXSncn3pcoFtKEirc{[teIFo\2WmIGPwbGsyKGW6cILld5Nm\CCrbjDiZYN2dG:4aYL1d{Bqdm[nY4Tl[EBU\jJzIHnud4VkfCClZXzsd{whU2l;MD6wNFM3|ryPLh?= M{jLN|xiKHSjcnfleF0oZ2KuYX7rK{BpemWoPTfoeJRxezpxL4D1Zo1m\C6wY3LpMo5tdS6waXiu[493NzJ6OEKyNlgyLz5{OEiyNlI5OTxxYU6=
HEK293 M2mwUmZ2dmO2aX;uJIF{e2G7 MWGzNEBucW6| MUjJcohq[mm2aX;uJI9nKEeIUD30ZYdo\WRiU1uxJEh2dmuwb4fuJI9zcWerbjmg[ZhxemW|c3XkJIlvKEiHS{K5N{Bk\WyuczD1d4lv\yCVcHigZZMhe3Wkc4TyZZRmKG2nYYP1doVlKGGodHXyJFMxKG2rboOgbY4heHKnc3XuZ4Uhd2ZiW3fhcY1iOzKSXVHUVEBjgSCFZYLlcotwfiClb4XueIlv\yCjbnHsfZNqeyxiSVO1NF0xNjB{ON88UU4> NETjR5Y9[SC2YYLn[ZQ:L1:kbHHub{chcHKnZk2nbJR1eHN8Lz;weYJu\WRwbnPibU5vdG1wbnnoModwfi9|MEi4PVM2Oid-M{C4PFk{PTJ:L3G+

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 Research:

[1]

  • Cell lines: 1483, A549, LN229, Jurkat, U937, MCF-7
  • Concentrations: ~1 μM
  • Incubation Time: 7 days
  • Method:

    CellTiter-Glo Assay

Solubility (25°C)

In vitro

Chemical Information

Molecular Weight 502.07
Formula

C27H32ClNO4S

CAS No. 1706522-79-3
Storage 3 years -20°C powder
2 years -80°C in solvent
Smiles CC1=CC(=CC(=C1)OCC2=CC=C(C=C2)CN3CCCC3CO)CS(=O)(=O)C4=CC=CC=C4.Cl

In vivo Formulation Calculator (Clear solution)

Step 1: Enter information below (Recommended: An additional animal making an allowance for loss during the experiment)

mg/kg g μL

Step 2: Enter the in vivo formulation (This is only the calculator, not formulation. Please contact us first if there is no in vivo formulation at the solubility Section.)

% DMSO % % Tween 80 % ddH2O
%DMSO %

Calculation results:

Working concentration: mg/ml;

Method for preparing DMSO master liquid: mg drug pre-dissolved in μL DMSO ( Master liquid concentration mg/mL, Please contact us first if the concentration exceeds the DMSO solubility of the batch of drug. )

Method for preparing in vivo formulation: Take μL DMSO master liquid, next addμL PEG300, mix and clarify, next addμL Tween 80, mix and clarify, next add μL ddH2O, mix and clarify.

Method for preparing in vivo formulation: Take μL DMSO master liquid, next add μL Corn oil, mix and clarify.

Note: 1. Please make sure the liquid is clear before adding the next solvent.
2. Be sure to add the solvent(s) in order. You must ensure that the solution obtained, in the previous addition, is a clear solution before proceeding to add the next solvent. Physical methods such
as vortex, ultrasound or hot water bath can be used to aid dissolving.

Molarity Calculator

Mass Concentration Volume Molecular Weight

Tech Support

Answers to questions you may have can be found in the inhibitor handling instructions. Topics include how to prepare stock solutions, how to store inhibitors, and issues that need special attention for cell-based assays and animal experiments.

Handling Instructions

Tel: +1-832-582-8158 Ext:3
If you have any other enquiries, please leave a message.

* Indicates a Required Field

Please enter your name.
Please enter your email. Please enter a valid email address.
Please write something to us.
Tags: buy PF-543 | PF-543 supplier | purchase PF-543 | PF-543 cost | PF-543 manufacturer | order PF-543 | PF-543 distributor