5-Lipoxygenase-activating protein is the target of a quinoline class of leukotriene synthesis inhibitors.
An indole class of leukotriene synthesis inhibitors, exemplified by MK-886, which does not directly inhibit 5-lipoxygenase, has been shown to bind to an 18-kDa leukocyte membrane protein and to inhibit 5-lipoxygenase membrane translocation. It was demonstrated that the 18-kDa protein is necessary for the cellular activation of leukotriene synthesis and was named 5-lipoxygenase-activating protein (FLAP). We describe here a class of leukotriene synthesis inhibitors based on a quinoline structure, which is structurally distinct from MK-886. However, similar to MK-886, several quinolines are potent inhibitors of cellular leukotriene synthesis but are poor inhibitors of soluble 5-lipoxygenase. To determine whether FLAP is the protein target of leukotriene synthesis inhibitors of the quinoline class, we investigated the ability of these compounds to inhibit photoaffinity labeling of FLAP and to elute FLAP from indole affinity gels. The abilities of the quinoline inhibitors to interact with FLAP correlated well with their abilities to inhibit leukotriene synthesis in human polymorphonuclear leukocytes. L-674,573, a potent quinoline leukotriene synthesis inhibitor, inhibited indole photoaffinity labeling of FLAP in a concentration-dependent manner. In addition, L-674,573 selectively eluted FLAP from indole affinity gels, in contrast to L-671,480, a quinoline that was inactive as an inhibitor of leukotriene synthesis. When human leukocyte membranes were labeled with the indole photoaffinity probe [125I]L-669,083 and immunoprecipitated with a FLAP antibody, the labeling of FLAP was inhibited by L-674,573 but not by L-671,480. These results suggest a direct binding site for the quinoline leukotriene synthesis inhibitors on FLAP and provide further evidence for the essential role of FLAP in cellular leukotriene synthesis.
Evans
J F
JF
Department of Pharmacology, Merck Frosst Centre for Therapeutic Research, Quebec, Canada.
Lévillé
C
C
Mancini
J A
JA
Prasit
P
P
Thérien
M
M
Zamboni
R
R
Gauthier
J Y
JY
Fortin
R
R
Charleson
P
P
MacIntyre
D E
DE
D058927:5-Lipoxygenase-Activating Proteins; D000818:Animals; D001665:Binding Sites; D002352:Carrier Proteins / Q000378:metabolism*; D002846:Chromatography, Affinity; D005782:Gels; D006801:Humans; D007211:Indoles / Q000378:metabolism / Q000494:pharmacology; D007962:Leukocytes / Q000187:drug effects / Q000378:metabolism; D020024:Leukotriene Antagonists; D015289:Leukotrienes / Q000096:biosynthesis*; D008565:Membrane Proteins / Q000378:metabolism*; D011804:Quinolines / Q000494:pharmacology*; D051381:Rats; D011919:Rats, Inbred Strains; D002722:Chlorobenzenes
D058927:5-Lipoxygenase-Activating Proteins; D002352:Carrier Proteins; D005782:Gels; D007211:Indoles; D020024:Leukotriene Antagonists; D015289:Leukotrienes; D008565:Membrane Proteins; D011804:Quinolines; C548592:ALOX5AP protein, human; C060893:MK-886; D002722:Chlorobenzenes