Benzopyrylium compounds

Patent metadata

Field	Value
Publication number	US-2015322078-A1
Application number	US-201314411447-A
Country	US
Kind code	A1
Filing date	Aug 20, 2013
Priority date	Aug 28, 2012
Publication date	Nov 12, 2015
Grant date	—

Abstract

Compounds used as labels with properties comparable to known fluorescent compounds. The compounds are conjugated to proteins and nucleic acids for biological imaging and analysis. Synthesis of the compounds, formation and use of the conjugated compounds, and specific non-limiting examples of each are provided.

First claim

What is claimed is:

1 . A compound according to any of general formula Ia

wherein
each of R 1 , R 2 , R 3 , R 4 , R 5 , R 6 , R 11 , and R 12 is the same or different and is independently selected from H, SO 3 , Z, L-Z, a PEG group P-L-Z where P is an ethylene glycol group, a diethylene glycol group, or a polyethylene glycol group, where the polyethylene glycol group is (CH 2 CH 2 O) s , where s is an integer from 3-6 inclusive, a sulfonamide group -L-SO 2 NH—P-L-Z, a caboxamide group -L-CONH—P-L-Z, hydrogen, alkyl-, tert-alkyl, aryl-, carboxyaryl-, dicarboxyaryl, heteroaryl-, cycloalkyl-, heterocycloalkyl-, alkyloxy-, alkylmercapto- with alkyl and cycloalkyl including olefin linkage residues, aryloxy-, arylmercapto-, heteroaryloxy-, heteroarylmercapto-, hydroxy-, nitro-, a carboxylic acid, an amino group, or cyano residues; where L is a divalent linear (—(CH 2 ) t —, t=0 to 15), branched, or cyclic alkane group that can be substituted by at least one atom of oxygen, nitrogen, substituted nitrogen, and/or sulfur; where Z is H, CH 3 , alkyl group, sulfoalkyl, heteroalkyl group, NH 2 , —COO − , —COOH, —COSH, CO—NH—NH 2 , —COF, —COCl, —COBr, —COI, —COO-Su (succinimidyl/sulfosuccinimidyl), —COO-STP (4-sulfo-2,3,5,6-tetrafluorophenyl), —COO-TFP (2,3,5,6-tetrafluorophenyl), —COO-benzotriazole, —CO-benzotriazole, —CONR′—CO—CH 2 —I, —CONR′R″, —CONR′-biomolecule, —CONR′-L-COO − , —CONR′-L-COOH, —CONR′-L-COO-Su, —CONR′-L-COO-STP, —CONR′-L-COO-TFP, —CONR′-L-CONR″ 2 , —CONR′-L-CO-biomolecule, —CONR′-L-CO—NH—NH 2 , —CONR′-L-OH, —CONR′-L-O-phosphoramidite, —CONR′-L-CHO, —CONR′-L-maleimide, or —CONR′-L-NH—CO—CH 2 —I; each of R′ and R″ is selected from H, aliphatic group, or heteroaliphatic group, and the biomolecule is a protein, peptide, antibody, nucleotide, oligonucleotide, biotin, or hapten;
X is selected from —OH, —SH, —NH 2 , —NH—NH 2 , —F, —Cl, —Br, I, —NHS (hydroxysuccinimidyl/sulfosuccinimidyl), —O-TFP (2,3,5,6-tetrafluorophenoxy), —O-STP (4-sulfo-2,3,5,6-tetrafluorophenoxy), —O-benzotriazole, -benzotriazole, —NR-L-OH, —NR-L-O-phosphoramidite, —NR-L-SH, —NR-L-NH 2 , —NR-L-NH—NH 2 , —NR-L-CO 2 H, —NR-L-CO—NHS, —NR-L-CO-STP, —NR-L-CO-TFP, —NR-L-CO-benzotriazole, —NR-L-CHO, —NR-L-maleimide, or —NR-L-NH—CO—CH2-I, where R is —H or an aliphatic or heteroaliphatic group;
each of R 10 , R 13 , and R 14 is the same or different and is independently selected from aliphatic, heteroaliphatic, sulfoalkyl group, carboxyalkyl group, heteroaliphatic with terminal SO 3 , Z, L-Z, PEG group P-L-Z where P is an ethylene glycol group, a diethylene glycol group, or a polyethylene glycol group where the polyethylene glycol group is (CH 2 CH 2 O) s , where s is an integer from 3-6 inclusive, a sulfonamide group -L-SO 2 NH—P-L-Z, or a caboxamide group -L-CONH—P-L-Z;
each of R 7 and R 9 is the same or different and is independently hydrogen, aliphatic group, heteroaliphatic group, or PEG group P-L-Z where P is selected from an ethylene glycol group, a diethylene glycol group, or a polyethylene glycol group where the polyethylene glycol group is (CH 2 CH 2 O) s , where s is an integer from 3-6 inclusive; or R7 and R9 together form a cyclic structure where R3 and R4 are joined using a divalent structural element selected from —(CH 2 ) q —, —(CH 2 ) q O(CH 2 ) q′ —, —(CH 2 ) q S(CH 2 ) q′ —, —(CH 2 ) q CH═CH—, —OCH═CH— where each of q and q′ is the same or different and is a integer from 1 to 6 inclusive; and
R 8 is selected from hydrogen, alkyl, sulfoalkyl, fluorine, chlorine, bromine, and PEG group P-L-Z where P is selected from an ethylene glycol group, a diethylene glycol group, or a polyethylene glycol group, where the polyethylene glycol group is (CH 2 CH 2 O) s , where s is an integer from 3-6 inclusive;
each of R 1 and R 2 , R 2 and R 3 , R 3 and R 4 , R 5 and R 6 , R 5 and R 8 , R 9 and R 10 , R 11 and R 12 , or R 12 and R 13 may form one or more aliphatic, heteroaliphatic or aromatic rings, and where the resultant ring(s) is optionally substituted by at least one alkyl-, sulfoalkyl, tert-alkyl, aryl-, carboxyaryl-, dicarboxyaryl, heteroaryl-, cycloalkyl-, heterocycloalkyl-, alkyloxy-, alkylmercapto- with alkyl and cycloalkyl including olefin linkage residues, aryloxy-, arylmercapto-, heteroaryloxy-, heteroarylmercapto-, hydroxy-, nitro-, sulfonic acid, a carboxylic acid, an amino group, or cyano residues;
at least one of R 1 -R 14 optionally contains in addition to at least one PEG an additional solubilizing, ionizing, or ionized substituent selected from SO 3 − , PO 3 2− , CO 2 H, OH, NR 3 + , cyclodextrins or sugars providing hydrophilic characteristics; the substituents optionally linked to chromophore by an aliphatic or heteroaliphatic or cyclical spacer;
Kat is a number of Na + , K + , Ca 2+ , ammonia, or other cation(s) needed to compensate the negative charge(s); n is 0, 1, 2, or 3; o is an integer from 0 to 12 inclusive; and p is an integer from 1 to 6 inclusive.

2 . The compound of claim 1 wherein
when the compound is according to general formula Ia or IIa each of R2 and R3 is independently selected from sulfoalkyl or a PEG group P-L-Z; R5 is alkyl; R6 is t-butyl or an unsubstituted or substituted phenyl; R11 is sulfonic acid, carboxylic acid, or an amino group; each of R13 and R14 is independently selected from alkyl, sulfoalkyl, or a PEG group P-L-Z; and each of R1, R4, R7, R8, R9, and R12 is H;
when the compound is according to general formula Ib or IIb each of R2 and R3 is independently selected from sulfoalkyl or a PEG group P-L-Z; R5 is alkyl; R6 is t-butyl or an unsubstituted or substituted phenyl; R11 is sulfonic acid, carboxylic acid, or an amino group; each of R13 and R14 is independently selected from alkyl, sulfoalkyl, or a PEG group P-L-Z; and each of R1, R4, R7, R8, R9, and R12 is H;
when the compound is according to general formula Ic or IIc each of R2 and R3 is independently selected from sulfoalkyl or a PEG group P-L-Z; R5 is alkyl; R6 is t-butyl or an unsubstituted or substituted phenyl; R11 is sulfonic acid, carboxylic acid, or an amino group; each of R13 and R14 is independently selected from alkyl, sulfoalkyl, or a PEG group P-L-Z; and each of R1, R4, R7, R8, R9, and R12 is H; and
when the compound is according to general formula Id or IId each of R2 and R3 is independently selected from sulfoalkyl or a PEG group P-L-Z; R5 is alkyl; R6 is t-butyl or an unsubstituted or substituted phenyl; R11 is sulfonic acid, carboxylic acid, or an amino group; each of R13 and R14 is independently selected from alkyl, sulfoalkyl, or a PEG group P-L-Z; and each of R1, R4, R7, R8, R9, and R12 is H.

3 . The compound of claim 2 wherein at least one of R2 or R3 is sulfopropyl, R5 is methyl, and at least one of R13 or R14 is sulfopropyl.

4 . The compound of claim 1 wherein
when the compound is according to general formula Ie or IIe each of R2 and R3 is independently selected from sulfoalkyl or a PEG group P-L-Z; R5 is alkyl; R6 is t-butyl or an unsubstituted or substituted phenyl; R11 is sulfonic acid, carboxylic acid, or an amino group; R10 is alkyl, sulfoalkyl, or a PEG group P-L-Z; R14 is alkyl; and each of R1, R4, R7, R8, R9, and R12 is H;
when the compound is according to general formula If or IIf each of R2 and R3 is independently selected from sulfoalkyl or a PEG group P-L-Z; R5 is alkyl; R6 is t-butyl or an unsubstituted or substituted phenyl; R11 is sulfonic acid, carboxylic acid, or an amino group; R10 is alkyl, sulfoalkyl, or a PEG group P-L-Z; R14 i

Assignees

• Hermanson Greg
• Czerney Peter T
• Desai Surbhi
• Wenzel Matthias S
• Lehmann Frank G
• Nlend Marie Christine
• Dyomics Gmbh
• Pierce Biotechnology Inc

Inventors

• Hermanson Greg
• Czerney Peter T
• Desai Surbhi
• Wenzel Matthias S
• Lehmann Frank G
• Nlend Marie Christine

Classifications

• C09B23/04

  one >CH- group, e.g. cyanines, isocyanines, pseudocyanines · CPC title

• A61K49/0058

  Antibodies · CPC title

• C09B23/083

  five >CH- groups · CPC title

• C09B69/00

  Dyes not provided for by a single group of this subclass · CPC title

• C07D491/16

  Peri-condensed systems · CPC title