Cyanine compounds

Patent metadata

Field	Value
Publication number	US-10696653-B2
Application number	US-201815976491-A
Country	US
Kind code	B2
Filing date	May 10, 2018
Priority date	Mar 2, 2012
Publication date	Jun 30, 2020
Grant date	Jun 30, 2020

Abstract

Compounds used as labels with properties comparable to known fluorescent compounds. The compounds can be 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 selected from the group consisting of

wherein each of R 1 , R 2 , R 5 , and R 6 is the same or different and is independently selected from the group consisting of aliphatic, heteroaliphatic, sulfoalkyl, heteroaliphatic with terminal SO 3 —, a PEG group P-L-Z where P is selected from an ethylene glycol group, a diethylene glycol group, and a (poly)ethylene glycol group where the (poly)ethylene 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, and a carboxamide group -L-CONH—P-L-Z;
each of R 7 and R 8 is the same or different and is independently selected from H, SO 3 —, a PEG group P-L-Z where P is selected from an ethylene glycol group, a diethylene glycol group, and a (poly)ethylene glycol group where the (poly)ethylene glycol group is (CH 2 CH 2 O) s where s is an integer from 3-6 inclusive, a sulfonamide group —SO 2 NH—P-L-Z, or a carboxamide group —CONH—P-L-Z;
where L is selected from the group consisting of a divalent linear (—(CH 2 ) o —, o=0 to 15), crossed, or cyclic alkane group that can be substituted by at least one atom selected from the group consisting of oxygen, substituted nitrogen, and/or sulfur;
where Z is selected from the group consisting of H, CH 3 , alkyl, heteroalkyl, NH 2 , —COO., —COOH, —COSH, CO—NH—NH 2 , —COF, —COCl, —COBr, —COI, —COO-Su (succinimidyl/sulfo-succinimidyl), —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, and —CONR′-L-NH—CO—CH 2 —I; R′ and R″ is selected from the group consisting of —H, aliphatic group, and heteroaliphatic group, and the biomolecule is a protein, antibody, nucleotide, oligonucleotide, biotin, or hapten;
X is selected from the group consisting of —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, and —NR-L-NH—CO—CH 2 —I, where R is —H, aliphatic, or heteroaliphatic;
Kat is a number of Na + , K + , Ca 2+ , ammonia, or other cation(s) needed to compensate the negative charge brought by the cyanine; m is an integer from 0 to 5 inclusive; p is an integer from 1 to 6 inclusive;
each of R 3 and R 4 is the same or different and is independently hydrogen, an aliphatic group, a heteroaliphatic group, or a PEG group P-L-Z where P is selected from an ethylene glycol group, a diethylene glycol group, and a (poly)ethylene glycol group where the (poly)ethylene glycol group is (CH 2 CH 2 O) s where s is an integer from 3-6 inclusive; or R 3 and R 4 together form a cyclic structure where R 3 and R 4 are joined using a divalent structural element selected from the group consisting of —(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 2 to 6 inclusive; and
Y is selected from the group consisting of —H, alkyl, sulfoalkyl, fluorine, chlorine, bromine, a PEG group P-L-Z where P is selected from an ethylene glycol group, a diethylene glycol group, and a (poly)ethylene glycol group where the (poly)ethylene glycol group is (CH 2 CH 2 O) s where s is an integer from 3-6 inclusive, and an oxygen-containing group OR PM where R PM is selected from the group consisting of hydrogen, substituted or unsubstituted alkyl, substituted or unsubstituted heteroalkyl, substituted or unsubstituted cyclic alkyl, substituted or unsubstituted heterocyclic alkyl, substituted or unsubstituted aryl, and substituted or unsubstituted heteroaryl, where the group is optionally substituted at least once with at least one of hydroxyl, sulfo, carboxy, and/or amino; with the proviso that at least one of R 1 and R 5 is a PEG group P-L-Z, and at least one of R 2 and R 6 is a PEG group P-L-Z.

2. A method of labeling at least one biomolecule, the method comprising combining at least one biomolecule with a composition comprising at least one excipient and the compound of claim 1 in an effective concentration to label at least one biomolecule under conditions sufficient for labeling the biomolecule with the compound.

3. The method of claim 2 wherein the biomolecule is selected from the group consisting of a protein, antibody, enzyme, nucleoside triphosphate, oligonucleotide, biotin, hapten, cofactor, lectin, antibody binding protein, carotenoid, carbohydrate, hormone, neurotransmitter, growth factors, toxin, biological cell, lipid, receptor binding drug, fluorescent proteins, organic polymer carrier material, inorganic carrier material, and combinations thereof.

4. A method of detecting at least one biomolecule, the method comprising combining at least one biomolecule with a composition comprising at least one excipient and the compound of claim 1 in an effective concentration to detect at least one biomolecule under conditions sufficient for binding the compound to the biomolecule, and detecting the biomolecule-bound compound.

5. The method of claim 4 wherein the biomolecule is selected from a protein, antibody, enzyme, nucleoside triphosphate, oligonucleotide, biotin, hapten, cofactor, lectin, antibody binding protein, carotenoid, carbohydrate, hormone, neurotransmitter, growth factors, toxin, biological cell, lipid, receptor binding drug, fluorescent proteins, organic polymer carrier material, inorganic carrier material, and combinations thereof.

6. The method of claim 4 wherein the at least one biomolecule is detected in an assay selected from fluorescence microscopy, flow cytometry, immunoassay, hybridization, chromatographic assay, electrophoretic assay, microwell plate based assay, fluorescence resonance energy transfer (FRET) system, bioluminescence resonance energy transfer (BRET) system, high throughput screening, or microarray.

7. The method of claim 4 wherein the biomolecule is detected by in vivo imaging comprising administering the biomolecule-bound compound to at least one of a biological sample, tissue, or organism, and detecting the biomolecule within the at least one of a biological sample, tissue, or organism.

8. A kit for labeling and/or detecting at least one biomolecule in a sample, the kit comprising the compound of claim 1 and at least one excipient, and instructions for use of the compound to label and/or detect a biomolecule in a sample.

Assignees

• Pierce Biotechnology Inc
• Dyomics Gmbh

Inventors

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

Classifications

• C09B23/0066

  the polymethine chain being part of a carbocyclic ring,(e.g. benzene, naphtalene, cyclohexene, cyclobutenene-quadratic acid) · CPC title

• C09B23/0025

  the substituent being bound through an oxygen atom · CPC title

• C09B23/0016

  the substituent being a halogen atom · CPC title

• C09B69/00

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

• C09B23/086

  more than five >CH- groups · CPC title