Modified Nucleic Acid Binding Cyanine Dyes for the Detection of Reactive Oxygen Species

摘要

Disclosed herein are compounds, compositions, methods, and kits for detecting reactive oxygen species (ROS) by conventional fluorescence microscopy, fluorescence spectroscopy, flow cytometry, and/or high content imaging. The compounds disclosed herein are novel reduced nucleic acid binding cyanine dyes, which dyes are probes for detecting ROS and measuring oxidative stress in cells either in vitro and/or in vivo. Also described herein are processes for preparing novel reduced dyes, i.e., ROS probes, for use in the disclosed compositions, methods and kits.

主权项

1. A reduced dye compound having structural formula (I): wherein each R1, which may be the same or different, is independently H; C1-C6 alkyl; trifluoromethyl; halogen; —OR8, —SR8, or —(NR8R9), where R8 and R9, which may be the same or different, are independently H; C1-C6 alkyl; or 1-2 alicyclic hetroalicyclic, aromatic or heteroaromatic rings having a total of 3-16 ring atoms, wherein the heteroatoms are O, N, or S; or R8 and R9 taken in combination are —(CH2)2-L-(CH2)2— where L is a single bond; —CH2—; —O—; or —NR10, where R10 is H or a C1-C6 alkyl;
t is 1, 2, 3 or 4; R2 is a saturated or unsaturated C1-C6 alkyl; X is O; S; Se; NR15, where R15 is H or a C1-C6 alkyl; or CR16R17, where R16 and R17, which may be the same or different, are independently H or C1-C6 alkyl, or the carbons of R16 and R17 taken in combination complete a five- or six-membered saturated ring; Q has the formula Q1 or Q2: whereinY is —CR3═CR4—;
p and m are 0 or 1, such that p+m=1;R5 is a C1-C6 alkyl, C1-C6 alkenyl, C1-C6 polyalkenyl, C1-C6 alkynyl or C1-C6 polyalkynyl group; or an OMEGA;R3, R4, R6 and R7, which may be the same or different, are each independently H; halogen; a C1-C6 alkyl, C1-C6 alkenyl, C1-C6 polyalkenyl, C1-C6 alkynyl or C1-C6 polyalkynyl group; —OR8; —SR8; —(NR8R9), as defined previously; —OSO2R19, where R19 is a C1-C6 alkyl, C1-C6 perfluoroalkyl, or aryl; or an OMEGA; or R6 and R7, taken in combination are —(CH2)v—, where v is 3 or 4, or R6 and R7 form a fused aromatic ring according to the formula Q2;R11, R12, R13 and R14, which may be the same or different, are each independently H; a C1-C6 alkyl, C1-C6 alkenyl, C1-C6 polyalkenyl, C1-C6 alkynyl or C1-C6 polyalkynyl group; a halogen; an OMEGA; or —OH, —OR8, —SR8, or —(NR8R9), as defined previously;OMEGA is a saturated or unsaturated, substituted or unsubstituted, cyclic substituent that has a total of 2-16 ring carbon atoms in 1-2 alicyclic, heteroalicyclic, aromatic, or heteroaromatic rings, containing 1-4 heteroatoms wherein the heteroatoms are O, N, or S, that is unsubstituted or optionally substituted one or more times, independently, by halogen, alkyl, perfluoroalkyl, amino, alkylamino, dialkylamino, alkoxy or carboxyalkyl, having 1-6 carbons, and that is attached as R3, R4, R5, R11, R12, R13, or R14 by a single bond, and, where more than one of R3, R4, R5, R11, R12, R13 and R14 is an OMEGA, each OMEGA is optionally the same or different; R20, R21 and R22 are independently H, C1-C6 alkyl, C3-C10 cycloalkyl, aryl or heteroaryl; W is H or D in either the R or S configuration; and n is 0, 1, 2, or 3.