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US 6,207,464 B1Granted
Rigidized monomethine cyanines
AssigneeCarnegie Mellon University
InventorsBhalchandra M. Karandikar, Alan S. Waggoner, Ratnakar B. Mujumdar
Life of this patent
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Abstract
Boron complexes of certain bis-heterocyclic compounds are provided. The complexes resemble monomethine cyanines and are useful for imparting fluorescent properties to materials by covalent and noncovalent association. The compounds have the following general formula: wherein the dotted lines Z1 and Z2 represent the atoms necessary to complete a structure selected from the group consisting of one ring, two fused rings, and three fused rings, each said ring having five or six atoms, and each said ring comprising carbon atoms and, optionally, no more than two atoms selected from oxygen, nitrogen and sulfur, and R1 through R5 represent various atoms or groups and M is Cl or F.
Claims26
A method for imparting fluorescent properties to a target, the method comprising incubating
- a target having at least one functional group selected from the group consisting of amino, hydroxyl, phosphoryl, carbonyl, and sulfhydryl groups, and
- an amount of a fluorescent molecule having the structure
- wherein
- X and Y are selected from the group consisting of —C(CH3)2—, oxygen, sulfur, —CH═CH—, and >N—W—K where N is nitrogen;
- dotted lines Z1 and Z2 represent the atoms necessary to complete a structure selected from the group consisting of one ring, two fused rings, and three fused rings, each said ring having five or six atoms, and each said ring comprising carbon atoms and, optionally, no more than two atoms selected from oxygen, nitrogen and sulfur;
- R1, R2, R3, R4 and R5 are selected from the group consisting of —K and —W—K;
- M is selected from the group consisting of F and Cl;
- W is a linker selected from the group consisting of branched alkyl chains of 1-27 carbon atoms, straight alkyl chains of 1-27 carbon atoms, monoethers containing 2-20 carbon atoms, and polyethers containing 2-20 carbon atoms;
- and
- K is a group conferring desired properties and is selected from the group consisting of:
- neutral groups that reduce water solubility selected from the group consisting of hydrogen and halogen atoms;
- polar groups that increase water solubility selected from the group consisting of amide, sulfonate, sulfate, phosphate, quaternary ammonium, hydroxyl and phosphonate;
- functional groups that can be used in labeling reactions selected from the group consisting of amino, hydroxyl, sulfhydryl, carboxyl and carbonyl;
- reactive groups selected from the group consisting of succinimidyl ester, isothiocyanate, isocyanate, iodoacetamide, maleimide, sulfonyl halide, phosphoramidite, alkylimidate, a arylimidate, acid halide, substituted hydrazines, substituted hydroxylamines, carbodiimides;
- and
- electron donating and withdrawing groups that shift the absorption and emission wavelengths of the fluorescent molecule;
- for a period of time sufficient to permit the fluorescent molecule to covalently bind to the at least one functional group of the target.
14 more elements
A method for imparting fluorescent properties to a target, the method comprising incubating
- an amount of a fluorescent molecule having the structure
- wherein:
- X and Y are selected from the group consisting of —C(CH3)2—, oxygen, sulfur, —CH═CH—, and >N—W—K where N is nitrogen;
- dotted lines Z1 and Z2 represent the atoms necessary to complete a structure selected from the group consisting of one ring, two fused rings, and three fused rings, each said ring having five or six atoms, and each said ring comprising carbon atoms and, optionally, no more than two atoms selected from oxygen, nitrogen and sulfur;
- R1, R2, R3, R4 and R5 are selected from the group consisting of —K and —W—K, and at least one of R1, R2, R3, R4 and R5 is a functional group selected from the group consisting of amino, hydroxyl, phosphoryl, carbonyl, and sulfhydryl;
- M is selected from the group consisting of F and Cl;
- W is a linker selected from the group consisting of branched alkyl chains of 1-27 carbon atoms, straight alkyl chains of 1-27 carbon atoms, monoethers containing 2-20 carbon atoms, and polyethers containing 2-20 carbon atoms;
- and
- K is a group conferring desired properties and is selected from the group consisting of:
- neutral groups that reduce water solubility selected from the group consisting of hydrogen and halogen atoms;
- polar groups that increase water solubility selected from the group consisting of amide, sulfonate, sulfate, phosphate, quaternary ammonium, hydroxyl and phosphonate;
- functional groups that can be used in labeling reactions selected from the group consisting of amino, hydroxyl, sulfhydryl, carboxyl and carbonyl;
- reactive groups selected from the group consisting of succinimidyl ester, isothiocyanate, isocyanate, iodoacetamide, maleimide, sulfonyl halide, phosphoramidite, alkylimidate, arylimidate, acid halide, substituted hydrazines, substituted hydroxylamines, carbodiimides;
- and
- electron donating and withdrawing groups that shift the absorption and emission wavelengths of the fluorescent molecule;
- and
- a target having at least one reactive group that can covalently bind with the at least one functional group, for a period of time sufficient to permit the at least one reactive group of the target to covalently bind to the at least one functional group of the fluorescent molecule.
14 more elements
A method of labeling a target, the method comprising incubating the target with an amount of a fluorescent molecule under conditions to form a covalent linkage between the target and the fluorescent molecule, the fluorescent molecule having the structure
- wherein:
- X and Y are selected from the group consisting of —C(CH3)2—, oxygen, sulfur, —CH═CH—, and >N—W—K where N is nitrogen;
- dotted lines Z1 and Z2 represent the atoms necessary to complete a structure selected from the group consisting of one ring, two fused rings, and three fused rings, each said ring having five or six atoms, and each said ring comprising carbon atoms and, optionally, no more than two atoms selected from oxygen, nitrogen and sulfur;
- R1, R2, R3, R4 and R5 are selected from the group consisting of —K and —W—K;
- M is selected from the group consisting of F and Cl;
- W is a linker selected from the group consisting of branched alkyl chains of 1-27 carbon atoms, straight alkyl chains of 1-27 carbon atoms, monoethers containing 2-20 carbon atoms, and polyethers containing 2-20 carbon atoms;
- and
- K is a group conferring desired properties and is selected from the group consisting of:
- neutral groups that reduce water solubility selected from the group consisting of hydrogen and halogen atoms;
- polar groups that increase water solubility selected from the group consisting of amide, sulfonate, sulfate, phosphate, quaternary ammonium, hydroxyl and phosphonate;
- functional groups that can be used in labeling reactions selected from the group consisting of amino, hydroxyl, sulfhydryl, carboxyl and carbonyl;
- reactive groups selected from the group consisting of succinimidyl ester, isothiocyanate, isocyanate, iodoacetamide, maleimide, sulfonyl halide, phosphoramidite, alkylimidate, arylimidate, acid halide, substituted hydrazines, substituted hydroxylamines, carbodiimides;
- and
- electron donating and withdrawing groups that shift the absorption and emission wavelengths of the fluorescent molecule.
11 more elements
A method of imparting fluorescent properties to a charged target, the method comprising incubating the target with an amount of a fluorescent molecule having the structure
- wherein:
- X and Y are selected from the group consisting of —C(CH3)2—, oxygen, sulfur, —CH═CH—, and >N—W—K where N is nitrogen;
- dotted lines Z1 and Z2 represent the atoms necessary to complete a structure selected from the group consisting of one ring, two fused rings, and three fused rings, each said ring having five or six atoms, and each said ring comprising carbon atoms and, optionally, no more than two atoms selected from oxygen, nitrogen and sulfur;
- R1, R2, R3, R4 and R5 are selected from the group consisting of —K and —W—K, and at least one of R1 through R5 is a charged group;
- M is selected from the group consisting of F and Cl;
- W is a linker selected from the group consisting of branched alkyl chains of 1-27 carbon atoms, straight alkyl chains of 1-27 carbon atoms, monoethers containing 2-20 carbon atoms, and polyethers containing 2-20 carbon atoms;
- and
- K is a group conferring desired properties and is selected from the group consisting of:
- neutral groups that reduce water solubility selected from the group consisting of hydrogen and halogen atoms;
- polar groups that increase water solubility selected from the group consisting of amide, sulfonate, sulfate, phosphate, quaternary ammonium, hydroxyl and phosphonate;
- functional groups that can be used in labeling reactions selected from the group consisting of amino, hydroxyl, sulfhydryl, carboxyl and carbonyl;
- reactive groups selected from the group consisting of succinimidyl ester, isothiocyanate, isocyanate, iodoacetamide, maleimide, sulfonyl halide, phosphoramidite, alkylimidate, arylimidate, acid halide, substituted hydrazines, substituted hydroxylamines, carbodiimides;
- and
- electron donating and withdrawing groups that shift the absorption and emission wavelengths of the fluorescent molecule;
- under conditions to noncovalently associate the fluorescent molecule with the target.
12 more elements
A method for detecting a target in a medium, the method comprising:
- (i) incubating a primary component having an affinity for the target with an amount of a fluorescent molecule having the structure
- wherein:
- X and Y are selected from the group consisting of —C(CH3)2—, oxygen, sulfur, —CH═CH—, and >N—W—K where N is nitrogen;
- dotted lines Z1 and Z2 represent the atoms necessary to complete a structure selected from the group consisting of one ring, two fused rings, and three fused rings, each said ring having five or six atoms, and each said ring comprising carbon atoms and, optionally, no more than two atoms selected from oxygen, nitrogen and sulfur;
- R1, R2, R3, R4 and R5 are selected from the group consisting of —K and —W—K;
- M is selected from the group consisting of F and Cl;
- W is a linker selected from the group consisting of branched alkyl chains of 1-27 carbon atoms, straight alkyl chains of 1-27 carbon atoms, monoethers containing 2-20 carbon atoms, and polyethers containing 2-20 carbon atoms;
- and
- K is a group conferring desired properties and is selected from the group consisting of:
- neutral groups that reduce water solubility selected from the group consisting of hydrogen and halogen atoms;
- polar groups that increase water solubility selected from the group consisting of amide, sulfonate, sulfate, phosphate, quaternary ammonium, hydroxyl and phosphonate;
- functional groups that can be used in labeling reactions selected from the group consisting of amino, hydroxyl, sulfhydryl, carboxyl and carbonyl;
- reactive groups selected from the group consisting of succinimidyl ester, isothiocyanates, isocyanate, iodoacetamide, maleimide, sulfonyl halide, phosphoramidite, alkylimidate, arylimidate, acid halide, substituted hydrazines, substituted hydroxylamines, carbodiimides;
- and
- electron donating and withdrawing groups that shift the absorption and emission wavelengths of the fluorescent molecule;
- under conditions to form a covalent linkage between the primary component and the flourescent molecule to thereby provide a fluorescently labeled primary component;
- and
- (ii) incubating an amount of the fluorescently labeled primary component with the medium under conditions to bind at least a portion of the flourescently labeled primary component with the target to thereby provide a fluorescently labeled target.
15 more elements
A method for quantifying a plurality of targets in a medium, the method comprising:
- (i) incubating at least a portion of the medium with an amount of a fluorescent molecule having the structure
- wherein:
- X and Y are selected from the group consisting of —C(CH3)2—, oxygen, sulfur, —CH═CH—, and >N—W—K where N is nitrogen;
- dotted lines Z1 and Z2 represent the atoms necessary to complete a structure selected from the group consisting of one ring, two fused rings, and three fused rings, each said ring having five or six atoms, and each said ring comprising carbon atoms and, optionally, no more than two atoms selected from oxygen, nitrogen and sulfur;
- R1, R2, R3, R4 and R5 are selected from the group consisting of —K and —W—K;
- M is selected from the group consisting of F and Cl;
- W is a linker selected from the group consisting of branched alkyl chains of 1-27 carbon atoms, straight alkyl chains of 1-27 carbon atoms, monoethers containing 2-20 carbon atoms, and polyethers containing 2-20 carbon atoms;
- and
- K is a group conferring desired properties and is selected from the group consisting of:
- neutral groups that reduce water solubility selected from the group consisting of hydrogen and halogen atoms;
- polar groups that increase water solubility selected from the group consisting of amide, sulfonate, sulfate, phosphate, quaternary ammonium, hydroxyl and phosphonate;
- functional groups that can be used in labeling reactions selected from the group consisting of amino, hydroxyl, sulfhydryl, carboxyl and carbonyl;
- reactive groups selected from the group consisting of succinimidyl ester, isothiocyanate, isocyanate, iodoacetamide, maleimide, sulfonyl halide, phosphoramidite, alkylimidate, arylimidate, acid halide, substituted hydrazines, substituted hydroxylamines, carbodiimides;
- and
- electron donating and withdrawing groups that shift the absorption and emission wavelengths of the fluorescent molecule;
- under conditions to form a covalent linkage between the fluorescent molecule and the plurality of targets, thereby providing a plurality of fluorescently labeled targets;
- (ii) separating the plurality of fluorescently labeled targets into separate portions according to at least one property of the targets;
- and
- (iii) determining the intensity of fluorescence of one or more of the separate portions.
16 more elements
A method for quantifying a plurality of targets in a medium, the method comprising:
- incubating a primary component having an affinity for the targets with an amount of a fluorescent molecule having the structure
- wherein:
- X and Y are selected from the group consisting of —C(CH3)2—, oxygen, sulfur, —CH═CH—, and >N—W—K where N is nitrogen;
- dotted lines Z1 and Z2 represent the atoms necessary to complete a structure selected from the group consisting of one ring, two fused rings, and three fused rings, each said ring having five or six atoms, and each said ring comprising carbon atoms and, optionally, no more than two atoms selected from oxygen, nitrogen and sulfur;
- R1, R2, R3, R4 and R5 are selected from the group consisting of —K and —W—K;
- M is selected from the group consisting of F and Cl;
- W is a linker selected from the group consisting of branched alkyl chains of 1-27 carbon atoms, straight alkyl chains of 1-27 carbon atoms, monoethers containing 2-20 carbon atoms, and polyethers containing 2-20 carbon atoms;
- and
- K is a group conferring desired properties and is selected from the group consisting of:
- neutral groups that reduce water solubility selected from the group consisting of hydrogen and halogen atoms;
- polar groups that increase water solubility selected from the group consisting of amide, sulfonate, sulfate, phosphate, quaternary ammonium, hydroxyl and phosphonate;
- functional groups that can be used in labeling reactions selected from the group consisting of amino, hydroxyl, sulfhydryl, carboxyl and carbonyl;
- reactive groups selected from the group consisting of succinimidyl ester, isothiocyanate, isocyanate, iodoacetamide, maleimide, sulfonyl halide, phosphoramidite, alkylimidate, arylimidate, acid halide, substituted hydrazines, substituted hydroxylamines, carbodiimides;
- and
- electron donating and withdrawing groups that shift the absorption and emission wavelengths of the fluorescent molecule;
- under conditions to form a covalent linkage between the primary component and the fluorescent molecule to provide a fluorescently labeled primary component;
- (ii) incubating an amount of the fluorescently labeled primary component with at least a portion of the medium under conditions to bind at least a portion of the fluorescently labeled primary component with the plurality of targets to provide fluorescently labeled targets;
- and
- (iii) separating the fluorescently labeled targets into separate portions according to at least one property of the targets.
16 more elements
Dependencies are inferred from the claim text.