title: Modulation of cellular S1P levels with a novel, potent and specific inhibitor of sphingosine kinase-1.
abstract: SphK (sphingosine kinase) is the major source of the bioactive lipid and GPCR (G-protein-coupled receptor) agonist S1P (sphingosine 1-phosphate). S1P promotes cell growth, survival and migration, and is a key regulator of lymphocyte trafficking. Inhibition of S1P signalling has been proposed as a strategy for treatment of inflammatory diseases and cancer. In the present paper we describe the discovery and characterization of PF-543, a novel cell-permeant inhibitor of SphK1. PF-543 inhibits SphK1 with a K(i) of 3.6 nM, is sphingosine-competitive and is more than 100-fold selective for SphK1 over the SphK2 isoform. In 1483 head and neck carcinoma cells, which are characterized by high levels of SphK1 expression and an unusually high rate of S1P production, PF-543 decreased the level of endogenous S1P 10-fold with a proportional increase in the level of sphingosine. In contrast with past reports that show that the growth of many cancer cell lines is SphK1-dependent, specific inhibition of SphK1 had no effect on the proliferation and survival of 1483 cells, despite a dramatic change in the cellular S1P/sphingosine ratio. PF-543 was 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. PF-543 is the most potent inhibitor of SphK1 described to date and it will be useful for dissecting specific roles of SphK1-driven S1P signalling.
authors.affiliation: Pfizer Global Research and Development, St. Louis Laboratories, Chesterfield, MO 63017, USA.
authors.forename: Mark E
authors.initials: ME
authors.lastname: Schnute
authors.forename: Matthew D
authors.initials: MD
authors.lastname: McReynolds
authors.forename: Tom
authors.initials: T
authors.lastname: Kasten
authors.forename: Matthew
authors.initials: M
authors.lastname: Yates
authors.forename: Gina
authors.initials: G
authors.lastname: Jerome
authors.forename: John W
authors.initials: JW
authors.lastname: Rains
authors.forename: Troii
authors.initials: T
authors.lastname: Hall
authors.forename: Jill
authors.initials: J
authors.lastname: Chrencik
authors.forename: Michelle
authors.initials: M
authors.lastname: Kraus
authors.forename: Ciaran N
authors.initials: CN
authors.lastname: Cronin
authors.forename: Matthew
authors.initials: M
authors.lastname: Saabye
authors.forename: Maureen K
authors.initials: MK
authors.lastname: Highkin
authors.forename: Richard
authors.initials: R
authors.lastname: Broadus
authors.forename: Shinji
authors.initials: S
authors.lastname: Ogawa
authors.forename: Kristin
authors.initials: K
authors.lastname: Cukyne
authors.forename: Laura E
authors.initials: LE
authors.lastname: Zawadzke
authors.forename: Vincent
authors.initials: V
authors.lastname: Peterkin
authors.forename: Kaliapan
authors.initials: K
authors.lastname: Iyanar
authors.forename: Jeffrey A
authors.initials: JA
authors.lastname: Scholten
authors.forename: Jay
authors.initials: J
authors.lastname: Wendling
authors.forename: Hideji
authors.initials: H
authors.lastname: Fujiwara
authors.forename: Olga
authors.initials: O
authors.lastname: Nemirovskiy
authors.forename: Arthur J
authors.initials: AJ
authors.lastname: Wittwer
authors.forename: Marek M
authors.initials: MM
authors.lastname: Nagiec
mesh_terms: D045744:Cell Line, Tumor; D002463:Cell Membrane Permeability; D006801:Humans; D008246:Lysophospholipids / Q000097:blood / Q000378:metabolism*; D000432:Methanol; D010766:Phosphorylation; D017853:Phosphotransferases (Alcohol Group Acceptor) / Q000037:antagonists & inhibitors*; D011759:Pyrrolidines / Q000138:chemical synthesis / Q000378:metabolism / Q000494:pharmacology*; D013110:Sphingosine / Q000031:analogs & derivatives* / Q000097:blood / Q000378:metabolism; D013379:Substrate Specificity; D013450:Sulfones / Q000138:chemical synthesis / Q000378:metabolism / Q000494:pharmacology*; D000099157:Sphingosine Kinase
chemical_list: D008246:Lysophospholipids; D000432:Methanol; D017853:Phosphotransferases (Alcohol Group Acceptor); D011759:Pyrrolidines; D013110:Sphingosine; D013450:Sulfones; D000099157:Sphingosine Kinase; C573330:PF-543; C060506:sphingosine 1-phosphate