Method and reagent for measuring nitroreductase enzyme activity

Details Method and reagent for measuring nitroreductase enzyme activity Method and reagent for measuring nitroreductase enzyme activity

2005-05-24

2009-08-25

Weber, Jon P (Department: 1657)

Chemistry: molecular biology and microbiology

Measuring or testing process involving enzymes or...

Reexamination Certificate

active

07579140

ABSTRACT:
Disclosed are nitro-substituted squaraine reporter dyes and methods using such dyes for detecting nitroreductase enzyme activity and nitroreductase gene expression in cellular assays. The dyes are of the structure:in which Z1and Z2independently represent a phenyl or a naphthyl ring system; X and Y are selected from oxygen, sulphur, —CH═CH— and the group:R1and R2are selected from C1-C4alkyl, —(CH2)n—P, —{(CH2)2—O}p—R6and group W; where P is selected from COOR7, SO3−and OH, W is mono- or di-substituted nitrobenzyl, R6is methyl or ethyl, R7is selected from H, C1-C4alkyl and CH2OC(O)R8, where R8is methyl, or t-butyl, n is an integer from 1 to 10, and p is an integer from 1 to 3; R3and R4are selected from hydrogen, NO2, halogen, SO3−, C1-C4alkoxy and —(CH2)m—COOR7; where R7is hereinbefore defined and m is 0 or an integer from 1 to 5; R5is C1-C6alkyl optionally substituted with COOR7, SO3−, or OH; where R7is hereinbefore defined; and at least one of groups R1, R2, R3and R4comprises at least one NO2group. Also provided are methods for screening for a test agent whose effect upon nitroreductase enzyme activity and nitroreductase gene expression is to be determined.

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