Fluorogenic substrates for beta -lactamase and methods of use

摘要

Fluorogenic substrates of the general formula I in which one of X and Y is a fluorescent donor moiety and the other is a quencher (which may or may not re-emit); R' is selected from the group consisting of H, lower (i.e., alkyl of 1 to about 5 carbon atoms) and (CH2OH)nOH, in which n is 0 or an integer from 1 to 5; R'' is selected from the group consisting of H, physiologically acceptable metal and ammonium cations, -CHR2OCO(CH2)nCH3, -CHR2OCOC(CH3)3, acylthiomethyl, acyloxy-alpha-benzyl, delta-butyrolactonyl, methoxycarbonyloxymethyl, phenyl, methylsulphinylmethyl, beta-morpholinoethyl, dialkylaminoethyl, acyloxyalkyl, dialkylaminocarbonyloxymethyl and aliphatic, in which R2 is selected from the group consisting of H and lower alkyl; A is selected from the group consisting of S, O, SO, SO2 and CH2; and Z' and Z'' are linkers for the fluorescent donor and quencher moieties. The substrates are useful in conjunction with beta -lactamase as reporter gene in a wide range of assays, for example to determine protein localization or bacterial resistance.