Benzopyrylium compounds

Patent metadata

Field	Value
Publication number	US-10174045-B2
Application number	US-201715493208-A
Country	US
Kind code	B2
Filing date	Apr 21, 2017
Priority date	Aug 28, 2012
Publication date	Jan 8, 2019
Grant date	Jan 8, 2019

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 is optionally 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 − , —COON, —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′-L-COO − , —CONR′-L-COOH, —CONR′-L-COO-Su, —CONR′-L-COO-STP, —CONR′-L-COO-TFP, —CONR′-L-CONR″ 2 , —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 H, aliphatic group, or heteroaliphatic group;
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—CH 2 —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 contains at least one PEG and optionally 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,
wherein the compound is covalently coupled to a biomolecule via X in general formula Ia, Ic, Ie, Ig, IIa, IIc, IIe or IIg, or the terminal —OH group in general formula Ib, Id, If, Ih, IIb, IId, IIf or IIh.

2. The compound of claim 1 wherein
when the compound is according to general formula Ia or IIa each of R 2 and R 3 is independently selected from sulfoalkyl or a PEG group P-L-Z; R 5 is alkyl; R 6 is t-butyl or an unsubstituted or substituted phenyl; R 11 is sulfonic acid, carboxylic acid, or an amino group; each of R 13 and R 14 is independently selected from alkyl, sulfoalkyl, or a PEG group P-L-Z; and each of R 1 , R 4 , R 7 , R 8 , R 9 , and R 12 is H;
when the compound is according to general formula Ib or IIb each of R 2 and R 3 is independently selected from sulfoalkyl or a PEG group P-L-Z; R 5 is alkyl; R 6 is t-butyl or an unsubstituted or substituted phenyl; R 11 is sulfonic acid, carboxylic acid, or an amino group; each of R 13 and R 14 is independently selected from alkyl, sulfoalkyl, or a PEG group P-L-Z; and each of R 1 , R 4 , R 7 , R 8 , R 9 , and R 12 is H;
when the compound is according to general formula Ic or IIc each of R 2 and R 3 is independently selected from sulfoalkyl or a PEG group P-L-Z; R 5 is alkyl; R 6 is t-butyl or an unsubstituted or substituted phenyl; R 11 is sulfonic acid, carboxylic acid, or an amino group; each of R 13 and R 14 is independently selected from alkyl, sulfoalkyl, or a PEG group P-L-Z; and each of R 1 , R 4 , R 7 , R 8 , R 9 , and R 12 is H; and
when the compound is according to general formula Id or IId each of R 2 and R 3 is independently selected from sulfoalkyl or a PEG group P-L-Z; R 5 is alkyl; R 6 is t-butyl or an unsubstituted or substituted phenyl; R 11 is sulfonic acid, carboxylic acid, or an amino group; each of R 13 and R 14 is independently selected from alkyl, sulfoalkyl, or a PEG group P-L-Z; and each of R 1 , R 4 , R 7 , R 8 , R 9 , and R 12 is H.

3. The compound of claim 2 wherein at least one of R 2 or R 3 is sulfopropyl, R 5 is methyl, and at least one of R 13 or R 14 is sulfopropyl.

4. The compound of claim 1 wherein
when the compound is according to general formula Ie or Ile each of R 2 and R 3 is independently selected from sulfoalkyl or a PEG group P-L-Z; R 5 is alkyl; R 6 is t-butyl or an unsubstituted or substituted phenyl; R 11 is sulfonic acid, carboxylic acid, or an amino group; R 10 is alkyl, sulfoalkyl, or a PEG group P-L-Z; R 14 is alkyl; and each of R 1 , R 4 , R 7 , R 8 , R 9 , and R 12 is H;
when the compound is according to general formula

Assignees

• Pierce Biotechnology Inc
• Dyomics Gmbh

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

• A61K49/0032

  Methine dyes, e.g. cyanine dyes · CPC title

• C07D405/06

  linked by a carbon chain containing only aliphatic carbon atoms · CPC title

• A61K49/0021

  the fluorescent group being a small organic molecule · CPC title

• C07D491/147 Primary

  the condensed system containing one ring with oxygen as ring hetero atom and two rings with nitrogen as ring hetero atom · CPC title