Derivatives of 1,2-dihydro-7-hydroxyquinolines containing fused rings

A method of staining a sample, said method comprising steps:

• combining a solution with said sample, wherein said solution comprises a compound having formula
• wherein
• R1, R2, R6, R41, R42, and R46 are independently selected from the group consisting of hydrogen, cyano, halogen, carboxylic acid, sulfonic acid C1-C6 alkyl, C1-C6 alkoxy, aryl, heteroaryl, —L—Rx and —L—SC, wherein said alkyl or alkoxy is optionally substituted by carboxylic acid, sulfonic acid, or halogen and said aryl or heteroaryl is optionally substituted one or more times by C1-C6 alkyl, C1-C6 perfluoroalkyl, cyano, halogen, azido, carboxylic acid, sulfonic acid, or halomethyl;
• or R1 in combination with R2, or R41 in combination with R42, or both, forms a fused aromatic or heteroaromatic ring that is optionally sulfonated one or more times or said ring is substituted by —L—Rx;
• or —L—SC;
• R3, R4, R43, and R44 are independently selected from the group consisting of hydrogen, C1-C6 alkyl, an aromatic or heteroaromatic ring, L—Rx and —L—SC, wherein said alkyl is optionally substituted by carboxylic acid, sulfonic acid, amino, hydroxy, or halogen and said aromatic or heteroaromatic ring that is optionally substituted one or more times by C1-C6 alkyl, C1-C6 alkoxy, C1-C6 perfluoroalkyl, cyano, halogen, carboxylic acid, sulfonic acid, or halomethyl;
• or R2 in combination with R3, R42 in combination with R43, or R3 in combination with R4, or R43 in combination with R44, or any combination thereof, forms a 5- or 6-membered alicyclic ring;
• R5 and R45 are independently selected from the group consisting of hydrogen, methyl, carboxymethyl, C2-C6 alkyl, aryl, heteroaryl, —L—Rx and —L—SC, wherein said alkyl is optionally substituted by carboxylic acid, sulfonic acid, amino, or halogen and said aryl or heteroaryl is optionally substituted one or more times by C1-C6 alkyl, C1-C6 perfluoroalkyl, cyano, halogen, carboxylic acid, sulfonic acid, or halomethyl;
• or R4 in combination with R5, or R5 in combination with R6, or R44 in combination with R45, or R45 in combination with R46, or any combination thereof, forms a 5- or 6-membered alicyclic ring;
• wherein one of E, E′, and X′ and X is O, S, or NR8, provided that E and X or E′ and X′ are not both present;
• wherein R8 is independently selected from the group consisting hydrogen, methyl, carboxymethyl, C2-C6 alkyl, —L—Rx and —L—SC, wherein said alkyl is optionally substituted by carboxylic acid, sulfonic acid, amino, or halogen;
• Q is N or CR28, wherein R28 is independently selected from the group consisting of hydrogen, F, CN, carboxylic acid, a carboxylic acid ester of a C1-C6 alcohol, C1-C6 alkyl, —L—Rx and —L—SC, wherein said alkyl is optionally substituted by carboxylic acid, sulfonic acid, amino, or halogen;
• or R28 comprises a formula
• wherein R30, R31, R32, R33 and R34 are independently selected from the group consisting of hydrogen, F, Cl, Br, I, sulfonic acid, carboxylic acid, CN, nitro, hydroxy, azido, amino, hydrazino, C1-C18 alkyl, C1-C18 alkoxy, C1-C18 alkylthio, C1-C18 alkanoylamino, C1-C18 alkylaminocarbonyl, C2-C36 dialkylaminocarbonyl, C1-C18 alkyloxycarbonyl, C7-C18 arylcarboxamido, —L—Rx and —L—SC, wherein said alkyl or aryl portions of said R30, R31, R32, R33 and R34 are optionally substituted one or more times by substituents selected from the group consisting of F, Cl, Br, I, hydroxy, carboxylic acid, a carboxylic acid ester of a C1-C6 alcohol, sulfonic acid, amino, C1-C6 alkylamino, C2-C6 dialkylamino and C1-C6 alkoxy;
• or a pair of adjacent R30, and R31, R32, R33 and R34, subsituents when taken in combination, form a fused 6-membered aromatic ring that is optionally further substituted by carboxylic acid;
• and
• wherein L is a covalent linkage;
• Rx is a reactive group;
• and
• SC is a conjugated substance;
• b) illuminating said sample with a suitable light wavelength to yield a detectable optical response.

A kit for staining a sample, wherein said kit comprises

• a solution comprising a buffer and a compound having formula
• wherein R1, R2, R5, R41, R42 and R46 are independently selected from the group consisting of hydrogen, cyano, halogen, carboxylic acid, sulfonic acid C1-C6 alkyl, C1-C6 alkoxy, aryl, heteroaryl, —L—Rx and —L—SC, wherein said alkyl or alkoxy is optionally substituted by carboxylic acid, sulfonic acid, or halogen and said aryl or heteroaryl is optionally substituted one or more times by C1-C6 alkyl, C1-C6 perfluoroalkyl, cyano, halogen, azido, carboxylic acid, sulfonic acid, or halomethyl;
• or R1 in combination with R2, or R41 in combination with R42, or both, forms a fused aromatic or heteroaromatic ring that is optionally sulfonated one or more times, or said ring is substituted by —L—Rx or —L—SC;
• R3, R4, R43, and R44 are independently selected from the group consisting of hydrogen, C1-C6 alkyl, an aromatic ring, a heteroaromatic ring, L—Rx and —L—SC, wherein said alkyl is optionally substituted by carboxylic acid, sulfonic acid, amino, hydroxy, or halogen and said aromatic or heteroaromatic ring is optionally substituted one or more times by C1-C6 alkyl, C1-C6 alkoxy, C1-C6 perfluoroalkyl, cyano, halogen, carboxylic acid, sulfonic acid, or halomethyl;
• or R2 in combination with R3, or R42 in combination with R43, or R3 in combination with R4, or R43 in combination with R44, or any combination thereof, forms a 5- or 6-membered alicyclic ring;
• R5 and R45 are independently selected from the group consisting of hydrogen, methyl, carboxymethyl, C2-C6 alkyl, aryl, heteroaryl, —L—Rx and —L—SC wherein said alkyl is optionally substituted by carboxylic acid, sulfonic acid, amino, or halogen and said aryl or heteroaryl is optionally substituted one or more times by C1-C6 alkyl, C1-C 6 perfluoroalkyl, cyano, halogen, carboxylic acid, sulfonic acid, or halomethyl;
• or R4 in combination with R5, or R5 in combination with R6, or R44 in combination with R45, or R45 in combination with R46, or any combination thereof, forms a 5- or 6-membered alicyclic ring;
• wherein one of said E, E′, X′ and X is O, S, or NR8, provided that E and X or E′ and X′ are not both present;
• wherein R8 is independently selected from the group consisting of hydrogen, methyl, carboxymethyl, C2-C6 alkyl, —L—Rx and —L—SC, wherein said alkyl is optionally substituted by carboxylic acid, sulfonic acid, amino, or halogen;
• and
• Q is N or CR28, wherein R28 is independently selected from the group consisting of hydrogen, F, CN, carboxylic acid, a carboxylic acid ester of a C1-C6 alcohol, a C1-C6 alkyl, —L—Rx and —L—SC, wherein said alkyl is optionally substituted by carboxylic acid, sulfonic acid, amino, or halogen:
• or R28 comprises a formula
• wherein R30, R31, R32, R33 and R34 are independently selected from the group consisting of hydrogen, F, Cl, Br, I, sulfonic acid, carboxylic acid, CN, nitro, hydroxy, azido, amino, hydrazine, C1-C18 alkyl, C1-C18 alkoxy, C1-C18 alkylthio, C1-C16, alkanoylamino, C1-C18 alkylaminocarbonyl, C2-C36 dialkylaminocarbonyl, C1-C18 alkyloxycarbonyl, C7-C18 arylcarboxarmido, —L—Rx and —L—SC, wherein said alkyl or aryl portions of said R30, R31, R32, R33 and R34 are optionally substituted one or more times by substituents selected from the group consisting of F, Cl, Br, I, hydroxy, carboxylic acid, a carboxylic acid ester of a C1-C6 alcohol, sulfonic acid, amino, C1-C6 alkylamino, C2-C6 dialkylamino and C1-C6 alkoxy;
• or a pair of adjacent R30, R31, R32, R33 and R34 substituents when taken in combination, form a fused 6-membered aromatic ring that is optionally further substituted by carboxylic acid;
• and
• wherein L is a covalent linkage;
• Rx is a reactive group;
• and
• SC is a conjugated substance.