Patent ID: 8512971

Claim:
A method for assaying the lysophospholipase D (LysoPLD) activity of autotoxin (ATX) comprising the steps of: a) obtaining a solution having ATX present; b) adding a LysoPLD substrate comprising a lysophospholipid derivative labeled with a fluorescent group and a quencher group to said solution containing ATX, wherein said fluorescent group and quencher group are both active in the near infrared spectrum; and c) determining if LysoPLD activity is present in said solution by measuring the change in the fluorescent signal in said solution that is generated by the cleavage of said lysophospholipid derivative represented by the formula: wherein A represents O or S; B represents H, Na + , K + , Li + or an alkyl group having 1-20 carbon atoms; E represents O, CH 2 , CHF or CF 2 ; G represents H, OH, F, O-acyl or O-alkyl having 1-20 carbon atoms; K 1 and K 2 each represent H, F, alkyl having 1 to 20 carbon atoms, OH or CO 2 H; W is C(O), NH(CH 2 CH 2 O) q C(O) or (NHC(O)(CH 2 ) n ) p C(O), wherein q is an integer from 1 to 33; n is an integer from 1 to 12; and p is an integer from 1 to 20; X represents NH; Y represents O, S or NH; Z is C(O)(CH 2 ) n (C)NH, (CH 2 CH 2 O) q C(O)NH or (CH 2 ) n NH wherein n is an integer from 1 to 33; and R 1 and R 2 are a fluorescent group or a quencher, respectively, wherein the fluorescent group is selected from Texas Red, tetramethylrhodamine and nitrobenzoxadiazole (NBD) and the quencher is