Methods to increase the photostability of dyes

摘要

The presently disclosed subject matter provides dyes having an improved photostability, biosensors comprising such dyes, and methods of use thereof, including methods for detecting target molecules in a sample under test and for live-cell imaging. The dyes can include a binding member, including a biomolecule or fragments thereof, which can interact with target molecules of interest and can be specific to a given conformational state or covalent modification, e.g., phosphorylation, of the target molecule. The presently disclosed dyes can be used for detecting changes in the binding, conformational change, or posttranslational modification of the target molecule.

主权项

1. A dye of Formula (II); wherein:
m is an integer from 1 to 4;X2 is halogen;R1, R2, R3, R6, and R7 are each independently selected from the group consisting of H, alkyl, substituted alkyl, heteroalkyl, substituted heteroalkyl, cycloalkyl, substituted cycloalkyl, cycloheteroalkyl, substituted cycloheteroalkyl, aryl, substituted aryl, aralkyl, hydroxyl, alkoxyl, hydroxyalkyl, hydroxycycloalkyl, alkoxycycloalkyl, aminoalkyl, acyloxyl, alkylaminoalkyl, alkoxycarbonyl, alkyleneaminoalkyl, alkylenesulfate, —(CH2)p—SO3, —(CH2)p—N+(CH3)2—(CH2)pNCS, and —(CH2)p—NH—CO—CH2—X2; wherein X2 is halogen;R5 is selected from the group consisting of alkyl, substituted alkyl, cycloalkyl, substituted cycloalkyl, cycloheteroalkyl, substituted cycloheteroalkyl, aryl, substituted aryl, aralkyl, hydroxyl, alkoxyl, hydroxyalkyl, hydroxycycloalkyl, alkoxycycloalkyl, aminoalkyl, acyloxyl, alkylaminoalkyl, alkoxycarbonyl, alkyleneaminoalkyl, alkylenesulfate, —(CH2)p—SO3, —(CH2)p—N+CH3)2—(CH2)p—NCS, and —(CH2)p—NH—CO—CH2—X2; wherein X2 is halogen;R8 is selected from the group consisting of H, alkyl, substituted alkyl, hydroxyl, amino, nitro, alkoxyl, and hydroxyalkyl;Y1 is H;Y2 is selected from the group consisting of H, —NH2, —SO3−, —(CH2)tOH, —(CH2)tNH3+X2−; CH3CONH—, X2CH2CONH—, HO(CH2)2—S—CH2CONH—, SuOCOCH2OCH2CON(CH3)—, —CH2—NH—C(═O)—O—(CH3)3, —(CH2)t—NH2+—(CH2)s—SO3−, —(CH2)t—NH—C(═O)—(CH2)s—X2, a succinimidyl ester, a hydroxysuccinimide, a maleimide, a haloacetyl, a pyridyl disulfide, a hydrazine, an ethyldiethylamino propylcarbodiimide,  or a protecting group, including t-butoxycarbonyl and carbobenzoxy;andwherein each p, r, s and t is independently an integer from 1 to 8; and Su is succinimidyl ester;and salts thereof.