Methods of diagnostic image analysis using bioconjugates of metal complexes of nitrogen-containing macrocyclic ligands

Abstract

The present invention is directed to bioconjugates of complexes represented by the formula: wherein R, R′, R 1 , R′ 1 , R 2 , R′ 2 , R 3 , R′ 3 , R 4 , R′ 4 , R 5 , R′ 5 , R 6 , R′ 6 , R 7 , R′ 7 , R 8 , R′ 8 , R 9 , R′ 9 , M, X, Y, Z and n are defined herein for use as contrast agents in diagnostic imaging.

That which is claimed is:  
     
         1 . A method of magnetic resonance imaging comprising: 
 (a) administering to a human or non-human animal subject a contrast medium comprising a physiologically compatible complex represented by the formula:                          wherein R, R′, R 1 , R′ 1 , R 2 , R′ 2 , R 3 , R′ 3 , R 4 , R′ 4 , R 5 , R′ 5 , R 6 , R′ 6 , R 7 , R′ 7 , R 8 , R′ 8 , R 9  and R′ 9  independently represents alkyl, alkenyl, alkynyl, cycloalkyl, cycloalkenyl, cycloalkylalkyl, cycloalkylcycloalkyl, cycloalkenylalkyl, alkylcycloalkyl, alkenylcycloalkyl, alkylcycloalkenyl, alkenylcycloalkenyl, heterocyclic, aryl and aralkyl radicals and radicals attached to the α-carbon of α-amino acids; or R 1  or R′ 1  and R 2  or R′ 2 , R 3  or R′ 3  and R 4  or R′4, R 5  or R′ 5  and R 6  or R′ 6 , R 7  or R′ 7  and R 8  or R′ 8 , and R 9  or R′ 9  and R or R′ together with the carbon atoms to which they are attached independently form a saturated, partially saturated or unsaturated cyclic having 3 to 20 carbon atoms; or R or R′ and R 1  or R′ 1 , R 2  or R′ 2  and R 3  or R′ 3 , R 4  or R′ 4  and R 5  or R′ 5 , R 6  or R′ 6  and R 7  or R′ 7 , and R 8  or R′ 8  and R 9  or R′ 9  together with the carbon atoms to which they are attached independently form a nitrogen containing heterocycle having 2 to 20 carbon atoms provided that when the nitrogen containing heterocycle is an aromatic heterocycle which does not contain a hydrogen attached to the nitrogen, the hydrogen attached to the nitrogen in said formula, which nitrogen is also in the macrocycle and the R groups attached to the same carbon atoms of the macrocycle are absent; and combinations thereof;    wherein (1) one to five of the “R” groups are attached to biomolecules via a linker group, (2) one of X, Y and Z is attached to a biomolecule via a linker group, or (3) one to five of the “R” groups and one of X, Y and Z are attached to biomolecules via a linker group; and said biomolecules are independently selected from the group consisting of steroids, carbohydrates, fatty acids, amino acids, peptides, proteins, antibodies, vitamins, lipids, phospholipids, phosphates, phosphonates, nucleic acids, enzyme substrates, enzyme inhibitors and enzyme receptor substrates and said linker group is derived from a substituent attached to said “R” group or said X, Y and Z which is reactive with the biomolecule and is selected from the group consisting of —NH 2 , —NHR 10 , —SH, —OH, —COOH, —COOR 10 , —CONH 2 , —NCO, —NCS, —COOX″, alkenyl, alkynyl, halide, tosylate, mesylate, tresylate, triflate and phenol, wherein R 10  is alkyl, aryl or alkaryl and X″ is a halide; and    wherein X, Y and Z are ligands independently selected from the group consisting of halide, oxo, aquo, hydroxo, alcohol, phenol, dioxygen, peroxo, hydroperoxo, alkylperoxo, arylperoxo, ammonia, alkylamino, arylamino, heterocycloalkyl amino, heterocycloaryl amino, amine oxides, hydrazine, alkyl hydrazine, aryl hydrazine, nitric oxide, cyanide, cyanate, thiocyanate, isocyanate, isothiocyanate, alkyl nitrile, aryl nitrile, alkyl isonitrile, aryl isonitrile, nitrate, nitrite, azido, alkyl sulfonic acid, aryl sulfonic acid, alkyl sulfoxide, aryl sulfoxide, alkyl aryl sulfoxide, alkyl sulfenic acid, aryl sulfenic acid, alkyl sulfinic acid, aryl sulfinic acid, alkyl thiol carboxylic acid, aryl thiol carboxylic acid, alkyl thiol thiocarboxylic acid, aryl thiol thiocarboxylic acid, alkyl carboxylic acid, aryl carboxylic acid, urea, alkyl urea, aryl urea, alkyl aryl urea, thiourea, alkyl thiourea, aryl thiourea, alkyl aryl thiourea, sulfate, sulfite, bisulfate, bisulfite, thiosulfate, thiosulfite, hydrosulfite, alkyl phosphine, aryl phosphine, alkyl phosphine oxide, aryl phosphine oxide, alkyl aryl phosphine oxide, alkyl phosphine sulfide, aryl phosphine sulfide, alkyl aryl phosphine sulfide, alkyl phosphonic acid, aryl phosphonic acid, alkyl phosphinic acid, aryl phosphinic acid, alkyl phosphinous acid, aryl phosphinous acid, phosphate, thiophosphate, phosphite, pyrophosphite, triphosphate, hydrogen phosphate, dihydrogen phosphate, alkyl guanidino, aryl guanidino, alkyl aryl guanidino, alkyl carbamate, aryl carbamate, alkyl aryl carbamate, alkyl thiocarbamate, aryl thiocarbamate, alkylaryl thiocarbamate, alkyl dithiocarbamate, aryl dithiocarbamate, alkylaryl dithiocarbamate, bicarbonate, carbonate, perchlorate, chlorate, chlorite, hypochlorite, perbromate, bromate, bromite, hypobromite, tetrahalomanganate, tetrafluoroborate, hexafluoroantimonate, hypophosphite, iodate, periodate, metaborate, tetraaryl borate, tetra alkyl borate, tartrate, salicylate, succinate, citrate, ascorbate, saccharinate, amino acid, hydroxamic acid, thiotosylate, and anions of ion exchange resins, or the corresponding anions thereof, or X, Y and Z are independently attached to one or more of the “R” groups and n is an integer from 0 to 3;    M is a paramagnetic metal selected from the group consisting of metals having atomic numbers 21-29, 42-44 and 57-71; and a nontoxic, pharmaceutically acceptable carrier, adjuvant or vehicle; and    (b) generating a magnetic resonance image of at least a part of said subject.    
     
     
         2 . Method of  claim 1  wherein M is Mn(II) or Gd(III).  
     
     
         3 . Method of  claim 2  wherein M is Mn(II).  
     
     
         4 . Method of  claim 1  wherein 1 to 2 of the “R” groups are attached to biomolecules via a linker group and none of X, Y and Z is attached to a biomolecule via a linker group.  
     
     
         5 . Method of  claim 1  wherein one of X, Y and Z is attached to a biomolecule via a linker group and none of the “R” groups are attached to biomolecules via a linker group.  
     
     
         6 . Method of  claim 1  wherein a maximum of one “R” group attached to the carbon atoms of the macrocycle located between nitrogen atoms has a biomolecule attached via a linker group.  
     
     
         7 . Method of  claim 1  wherein at least one of the “R” groups, in addition to the “R” groups which are attached to biomolecules via a linker group, are independently selected from the group consisting of alkyl, cycloalkyl, cycloalkylalkyl, aralkyl, alkaryl, aryl, heterocyclics and radicals attached to the α-carbon of α-amino acids, and the remaining “R” groups are independently selected from hydrogen, saturated, partially saturated or unsaturated cyclics or a nitrogen containing heterocycle.  
     
     
         8 . Method of  claim 7  wherein at least two of the “R” groups, in addition to the “R” groups which are attached to biomolecules via a linker group, are independently selected from the group consisting of alkyl, cycloalkyl, cycloalkylalkyl, aralkyl, alkaryl, aryl, heterocyclics and radicals attached to the α-carbon of α-amino acids.  
     
     
         9 . Method of  claim 7  wherein at least one of the “R” groups, in addition to the “R” groups which are attached to biomolecules, via a linker group, are alkyl and the remaining “R” groups are independently selected from hydrogen or saturated, partially saturated or unsaturated cyclics.  
     
     
         10 . Method of  claim 1  wherein at least one of R 1  or R′ 1  and R 2  or R′ 2 , R 3  or R′ 3  and R 4  or R′ 4 , R 5  or R′5 and R 6  or R′ 6 , R 7  or R′ 7  and R 8  or R′ 8 , and R 9  or R′ 9  and R or R′ together with the carbon atoms to which they are attached represent a saturated, partially saturated or unsaturated cyclic having 3 to 20 carbon atoms and the remaining “R” groups in addition to the “R” groups which are attached to biomolecules via linker groups are independently selected from hydrogen, nitrogen containing heterocycles or alkyl groups.  
     
     
         11 . Method of  claim 10  wherein at least two of R 1  or R′ 1  and R 2  or R′ 2 , R 3  or R′ 3  and R 4  or R′ 4 , R 5  or R′ 5  and R 6  or R′ 6 , R 7  or R′ 7  and R 8  or R′ 8 , and R 9  or R′ 9  and R or R′ together with the carbon atoms to which they are attached represent a saturated, partially saturated or unsaturated cyclic having 3 to 20 carbon atoms and the remaining “R” groups in addition to the “R” groups which are attached to biomolecules via linker groups are independently selected from hydrogen, nitrogen containing heterocycles or alkyl groups.  
     
     
         12 . Method of  claim 10  wherein said saturated, partially saturated or unsaturated cyclic is cyclohexyl.  
     
     
         13 . Method of  claim 12  wherein said remaining “R” groups in addition to the “R” groups which are attached to biomolecules via linker groups are independently selected from hydrogen or alkyl groups.  
     
     
         14 . Method of  claim 1  wherein said R or R′ and R 1  or R 1 ′, R 2  or R 2 ′ and R 3  or R 3 ′, R 4  or R′ 4  and R 5  or R′ 5 , R 6  or R 6 ′ and R 7  or R 7 ′, and R 8  or R′ 8  and R 9  or R 9 ′ together with the carbon atoms to which they are attached are found to form a nitrogen containing heterocycle having 2 to 20 carbon atoms, and the remaining “R” groups in addition to the “R” groups which are attached to biomolecules via a linker group are independently selected from hydrogen, saturated, partially saturated or unsaturated cyclics or alkyl groups.  
     
     
         15 . Method of  claim 1  wherein X, Y and Z are independently selected from the group consisting of halide, organic acid, nitrate and bicarbonate anions.  
     
     
         16 . A method of diagnostic imaging comprising: 
 (a) administering to a human or non-human animal subject a diagnostic agent comprising a physiologically compatible complex represented by the formula:                          wherein R, R′, R 1 , R′ 1 , R 2 , R′ 2 , R 3 , R′ 3  R 4 , R′ 4 , R 5 , R′ 5 , R 6 , R′ 6 , R 7 , R′ 7 , R 8 , R′ 8 , R 9  and R′ 9  independently represents alkyl, alkenyl, alkynyl, cycloalkyl, cycloalkenyl, cycloalkylalkyl, cycloalkylcycloalkyl, cycloalkenylalkyl, alkylcycloalkyl, alkenylcycloalkyl, alkylcycloalkenyl, alkenylcycloalkenyl, heterocyclic, aryl and aralkyl radicals and radicals attached to the α-carbon of α-amino acids; or R 1  or R′ 1  and R 2  or R′ 2 , R 3  or R′ 3  and R 4  or R′ 4 , R 5  or R′ 5  and R 6  or R′ 16 , R 7  or R′ 7  and R 8  or R′ 8 , and R 9  or R′ 9  and R or R′ together with the carbon atoms to which they are attached independently form a saturated, partially saturated or unsaturated cyclic having 3 to 20 carbon atoms; or R or R′ and R 1  or R′ 1 , R 2  or R′ 2  and R 3  or R′ 3 , R 4  or R′ 4  and R 5  or R′ 5 , R 6  or R′ 6  and R 7  or R′ 7 , and R 8  or R′ 8  and R 9  or R′ 9  together with the carbon atoms to which they are attached independently form a nitrogen containing heterocycle having 2 to 20 carbon atoms provided that when the nitrogen containing heterocycle is an aromatic heterocycle which does not contain a hydrogen attached to the nitrogen, the hydrogen attached to the nitrogen in said formula, which nitrogen is also in the macrocycle and the R groups attached to the same carbon atoms of the macrocycle are absent; and combinations thereof;    wherein (1) one to five of the “R” groups are attached to biomolecules via a linker group, (2) one of X, Y and Z is attached to a biomolecule via a linker group, or (3) one to five of the “R” groups and one of X, Y and Z are attached to biomolecules via a linker group; and said biomolecules are independently selected from the group consisting of steroids, carbohydrates, fatty acids, amino acids, peptides, proteins, antibodies, vitamins, lipids, phospholipids, phosphates, phosphonates, nucleic acids, enzyme substrates, enzyme inhibitors and enzyme receptor substrates and said linker group is derived from a substituent attached to said “R” group or said X, Y and Z which is reactive with the biomolecule and is selected from the group consisting of —NH 2 , —NHR 10 , —SH, —OH, —COOH, —COOR 10 , —CONH 2 , —NCO, —NCS, —COOX″, alkenyl, alkynyl, halide, tosylate, mesylate, tresylate, triflate and phenol, herein R 10  is alkyl, aryl or alkaryl and X″ is a halide; and wherein X, Y and Z are ligands independently selected from the group consisting of halide, oxo, aquo, hydroxo, alcohol, phenol, dioxygen, peroxo, hydroperoxo, alkylperoxo, arylperoxo, ammonia, alkylamino, arylamino, heterocycloalkyl amino, heterocycloaryl amino, amine oxides, hydrazine, alkyl hydrazine, aryl hydrazine, nitric oxide, cyanide, cyanate, thiocyanate, isocyanate, isothiocyanate, alkyl nitrile, aryl nitrile, alkyl isonitrile, aryl isonitrile, nitrate, nitrite, azido, alkyl sulfonic acid, aryl sulfonic acid, alkyl sulfoxide, aryl sulfoxide, alkyl aryl sulfoxide, alkyl sulfenic acid, aryl sulfenic acid, alkyl sulfinic acid, aryl sulfinic acid, alkyl thiol carboxylic acid, aryl thiol carboxylic acid, alkyl thiol thiocarboxylic acid, aryl thiol thiocarboxylic acid, alkyl carboxylic acid, aryl carboxylic acid, urea, alkyl urea, aryl urea, alkyl aryl urea, thiourea, alkyl thiourea, aryl thiourea, alkyl aryl thiourea, sulfate, sulfite, bisulfate, bisulfite, thiosulfate, thiosulfite, hydrosulfite, alkyl phosphine, aryl phosphine, alkyl phosphine oxide, aryl phosphine oxide, alkyl aryl phosphine oxide, alkyl phosphine sulfide, aryl phosphine sulfide, alkyl aryl phosphine sulfide, alkyl phosphonic acid, aryl phosphonic acid, alkyl phosphinic acid, aryl phosphinic acid, alkyl phosphinous acid, aryl phosphinous acid, phosphate, thiophosphate, phosphite, pyrophosphite, triphosphate, hydrogen phosphate, dihydrogen phosphate, alkyl guanidino, aryl guanidino, alkyl aryl guanidino, alkyl carbamate, aryl carbamate, alkyl aryl carbamate, alkyl thiocarbamate, aryl thiocarbamate, alkylaryl thiocarbamate, alkyl dithiocarbamate, aryl dithiocarbamate, alkylaryl dithiocarbamate, bicarbonate, carbonate, perchlorate, chlorate, chlorite, hypochlorite, perbromate, bromate, bromite, hypobromite, tetrahalomanganate, tetrafluoroborate, hexafluoroantimonate, hypophosphite, iodate, periodate, metaborate, tetraaryl borate, tetra alkyl borate, tartrate, salicylate, succinate, citrate, ascorbate, saccharinate, amino acid, hydroxamic acid, thiotosylate, and anions of ion exchange resins, or the corresponding anions thereof, or X, Y and Z are independently attached to one or more of the “R” groups and n is an integer from 0 to 3; M is a heavy metal selected from the group consisting of metals having atomic numbers 20-32, 42-44, 49 and 57-83; and a nontoxic, pharmaceutically acceptable carrier, adjuvant or vehicle; and    (b) generating an X-ray, ultrasound or scintigraphic image of at least a part of said subject.    
     
     
         17 . Method of  claim 16  wherein M is a radioactive metal isotope selected from the group consisting of  99m Tc and  111 In and said image is a scintigraphic image.  
     
     
         18 . A method of radiotherapy practiced on a human or non-human animal subject comprising administering to said subject a radioactive agent comprising a physiologically compatible complex represented by the formula:  
       
         
           
           
               
               
           
         
       
       wherein R, R′, R 1 , R′ 1 , R 2 , R′ 2 , R 3 , R′ 3  R 4 , R′ 4 , R 5 , R′ 5 , R 6 , R′ 6 , R 7 , R′ 7 , R 8 , R′ 8 , R 9  and R′ 9  independently represents alkyl, alkenyl, alkynyl, cycloalkyl, cycloalkenyl, cycloalkylalkyl, cycloalkylcycloalkyl, cycloalkenylalkyl, alkylcycloalkyl, alkenylcycloalkyl, alkylcycloalkenyl, alkenylcycloalkenyl, heterocyclic, aryl and aralkyl radicals and radicals attached to the α-carbon of α-amino acids; or R 1  or R′ 1  and R 2  or R′ 2 , R 3  or R′ 3  and R 4  or R′ 4 , R 5  or R′ 5  and R 6  or R′ 6 , R 7  or R′ 7  and R 8  or R′ 8 , and R 9  or R′ 9  and R or R′ together with the carbon atoms to which they are attached independently form a saturated, partially saturated or unsaturated cyclic having 3 to 20 carbon atoms; or R or R′ and R 1  or R′ 1 , R 2  or R′ 2  and R 3  or R′ 3 , R 4  or R′ 4  and R 5  or R′ 5 , R 6  or R′ 6  and R 7  or R′ 7 , and R 8  or R′ 8  and R 9  or R′ 9  together with the carbon atoms to which they are attached independently form a nitrogen containing heterocycle having 2 to 20 carbon atoms provided that when the nitrogen containing heterocycle is an aromatic heterocycle which does not contain a hydrogen attached to the nitrogen, the hydrogen attached to the nitrogen in said formula, which nitrogen is also in the macrocycle and the R groups attached to the same carbon atoms of the macrocycle are absent; and combinations thereof; 
 wherein (1) one to five of the “R” groups are attached to biomolecules via a linker group, (2) one of X, Y and Z is attached to a biomolecule via a linker group, or (3) one to five of the “R” groups and one of X, Y and Z are attached to biomolecules via a linker group; and said biomolecules are independently selected from the group consisting of steroids, carbohydrates, fatty acids, amino acids, peptides, proteins, antibodies, vitamins, lipids, phospholipids, phosphates, phosphonates, nucleic acids, enzyme substrates, enzyme inhibitors and enzyme receptor substrates and said linker group is derived from a substituent attached to said “R” group or said X, Y and Z which is reactive with the biomolecule and is selected from the group consisting of —NH 2 , —NHR 10 , —SH, —OH, —COOH, —COOR 10 , —CONH 2 , —NCO, —NCS, —COOX″, alkenyl, alkynyl, halide, tosylate, mesylate, tresylate, triflate and phenol, wherein R 10  is alkyl, aryl or alkaryl and X″ is a halide; and  
 wherein X, Y and Z are ligands independently selected from the group consisting of halide, oxo, aquo, hydroxo, alcohol, phenol, dioxygen, peroxo, hydroperoxo, alkylperoxo, arylperoxo, ammonia, alkylamino, arylamino, heterocycloalkyl amino, heterocycloaryl amino, amine oxides, hydrazine, alkyl hydrazine, aryl hydrazine, nitric oxide, cyanide, cyanate, thiocyanate, isocyanate, isothiocyanate, alkyl nitrile, aryl nitrile, alkyl isonitrile, aryl isonitrile, nitrate, nitrite, azido, alkyl sulfonic acid, aryl sulfonic acid, alkyl sulfoxide, aryl sulfoxide, alkyl aryl sulfoxide, alkyl sulfenic acid, aryl sulfenic acid, alkyl sulfinic acid, aryl sulfinic acid, alkyl thiol carboxylic acid, aryl thiol carboxylic acid, alkyl thiol thiocarboxylic acid, aryl thiol thiocarboxylic acid, alkyl carboxylic acid, aryl carboxylic acid, urea, alkyl urea, aryl urea, alkyl aryl urea, thiourea, alkyl thiourea, aryl thiourea, alkyl aryl thiourea, sulfate, sulfite, bisulfate, bisulfite, thiosulfate, thiosulfite, hydrosulfite, alkyl phosphine, aryl phosphine, alkyl phosphine oxide, aryl phosphine oxide, alkyl aryl phosphine oxide, alkyl phosphine sulfide, aryl phosphine sulfide, alkyl aryl phosphine sulfide, alkyl phosphonic acid, aryl phosphonic acid, alkyl phosphinic acid, aryl phosphinic acid, alkyl phosphinous acid, aryl phosphinous acid, phosphate, thiophosphate, phosphite, pyrophosphite, triphosphate, hydrogen phosphate, dihydrogen phosphate, alkyl guanidino, aryl guanidino, alkyl aryl guanidino, alkyl carbamate, aryl carbamate, alkyl aryl carbamate, alkyl thiocarbamate, aryl thiocarbamate, alkylaryl thiocarbamate, alkyl dithiocarbamate, aryl dithiocarbamate, alkylaryl dithiocarbamate, bicarbonate, carbonate, perchlorate, chlorate, chlorite, hypochlorite, perbromate, bromate, bromite, hypobromite, tetrahalomanganate, tetrafluoroborate, hexafluoroantimonate, hypophosphite, iodate, periodate, metaborate, tetraaryl borate, tetra alkyl borate, tartrate, salicylate, succinate, citrate, ascorbate, saccharinate, amino acid, hydroxamic acid, thiotosylate, and anions of ion exchange resins, or the corresponding anions thereof, or X, Y and Z are independently attached to one or more of the “R” groups and n is an integer from 0 to 3;  
 M is a radioactive metal isotope selected from the group consisting of  153 Sm,  67 Cu and  90 Y; and a nontoxic, pharmaceutically acceptable carrier, adjuvant or vehicle.  
 
     
     
         19 . A compound which is a complex represented by the formula:  
       
         
           
           
               
               
           
         
       
       wherein R, R′, R 1 , R′ 1 , R 2 , R′ 2 , R 3 , R′ 3  R 4 , R′ 4 , R 5 , R′ 5 R 6 , R′ 6 , R 7 , R′ 7 , R 8 , R′ 8 , R 9  and R′ 9  independently represents alkyl, alkenyl, alkynyl, cycloalkyl, cycloalkenyl, cycloalkylalkyl, cycloalkylcycloalkyl, cycloalkenylalkyl, alkylcycloalkyl, alkenylcycloalkyl, alkylcycloalkenyl, alkenylcycloalkenyl, heterocyclic, aryl and aralkyl radicals and radicals attached to the α-carbon of α-amino acids; or R 1  or R′ 1  and R 2  or R′ 2 , R 3  or R′ 3  and R 4  or R′ 4 , R 5  or R′ 5  and R 6  or R′ 6 , R 7  or R′ 7  and R 8  or R′ 8 , and R 9  or R′ 9  and R or R′ together with the carbon atoms to which they are attached independently form a saturated, partially saturated or unsaturated cyclic having 3 to 20 carbon atoms; or R or R′ and R 1  or R′ 1 , R 2  or R′ 2  and R 3  or R′ 3 , R 4  or R′ 4  and R 5  or R′ 5 , R 6  or R′ 6  and R 7  or R′ 7 , and R 8  or R′ 8  and R 9  or R′ 9  together with the carbon atoms to which they are attached independently form a nitrogen containing heterocycle having 2 to 20 carbon atoms provided that when the nitrogen containing heterocycle is an aromatic heterocycle which does not contain a hydrogen attached to the nitrogen, the hydrogen attached to the nitrogen in said formula, which nitrogen is also in the macrocycle and the R groups attached to the same carbon atoms of the macrocycle are absent; and combinations thereof; 
 wherein (1) one to five of the “R” groups are attached to biomolecules via a linker group, (2) one of X, Y and Z is attached to a biomolecule via a linker group, or (3) one to five of the “R” groups and one of X, Y and Z are attached to biomolecules via a linker group; and said biomolecules are independently selected from the group consisting of steroids, carbohydrates, fatty acids, amino acids, peptides, proteins, antibodies, vitamins, lipids, phospholipds, phosphates, phosphonates, nucleic acids, enzyme substrates, enzyme inhibitors and enzyme receptor substrates and said linker group is derived from a substituent attached to aid “R” group or said X, Y and Z which is reactive with he biomolecule and is selected from the group consisting of —NH 2 , —NHR 10 , —SH, —OH, —COOH, —COOR 10 , —CONH 2 , —NCO, —NCS, —COOX″, alkenyl, alkynyl, halide, tosylate, mesylate, tresylate, triflate and phenol, wherein R 10  is alkyl, aryl or alkaryl and X″ is a halide; and  
 wherein X, Y and Z are ligands independently selected from the group consisting of halide, oxo, aquo, hydroxo, alcohol, phenol, dioxygen, peroxo, hydroperoxo, alkylperoxo, arylperoxo, ammonia, alkylamino, arylamino, heterocycloalkyl amino, heterocycloaryl amino, amine oxides, hydrazine, alkyl hydrazine, aryl hydrazine, nitric oxide, cyanide, cyanate, thiocyanate, isocyanate, isothiocyanate, alkyl nitrile, aryl nitrile, alkyl isonitrile, aryl isonitrile, nitrate, nitrite, azido, alkyl sulfonic acid, aryl sulfonic acid, alkyl sulfoxide, aryl sulfoxide, alkyl aryl sulfoxide, alkyl sulfenic acid, aryl sulfenic acid, alkyl sulfinic acid, aryl sulfinic acid, alkyl thiol carboxylic acid, aryl thiol carboxylic acid, alkyl thiol thiocarboxylic acid, aryl thiol thiocarboxylic acid, alkyl carboxylic acid, aryl carboxylic acid, urea, alkyl urea, aryl urea, alkyl aryl urea, thiourea, alkyl thiourea, aryl thiourea, alkyl aryl thiourea, sulfate, sulfite, bisulfate, bisulfite, thiosulfate, thiosulfite, hydrosulfite, alkyl phosphine, aryl phosphine, alkyl phosphine oxide, aryl phosphine oxide, alkyl aryl phosphine oxide, alkyl phosphine sulfide, aryl phosphine sulfide, alkyl aryl phosphine sulfide, alkyl phosphonic acid, aryl phosphonic acid, alkyl phosphinic acid, aryl phosphinic acid, alkyl phosphinous acid, aryl phosphinous acid, phosphate, thiophosphate, phosphite, pyrophosphite, triphosphate, hydrogen phosphate, dihydrogen phosphate, alkyl guanidino, aryl guanidino, alkyl aryl guanidino, alkyl carbamate, aryl carbamate, alkyl aryl carbamate, alkyl thiocarbamate, aryl thiocarbamate, alkylaryl thiocarbamate, alkyl dithiocarbamate, aryl dithiocarbamate, alkylaryl dithiocarbamate, bicarbonate, carbonate, perchlorate, chlorate, chlorite, hypochlorite, perbromate, bromate, bromite, hypobromite, tetrahalomanganate, tetrafluoroborate, hexafluoroantimonate, hypophosphite, iodate, periodate, metaborate, tetraaryl borate, tetra alkyl borate, tartrate, salicylate, succinate, citrate, ascorbate, saccharinate, amino acid, hydroxamic acid, thiotosylate, and anions of ion exchange resins, or the corresponding anions thereof, or X, Y and Z are independently attached to one or more of the “R” groups and n is an integer from 0 to 3; and  
 M is a paramagnetic metal selected from the group consisting of metals having atomic numbers 21-24, 26-29, 42-44, 49 and 57-83.  
 (b) generating a magnetic resonance image of at least a part of said subject.  
 
     
     
         20 . Compound of  claim 19  wherein M is Gd(III).  
     
     
         21 . Compound of  claim 19  wherein 1 to 2 of the “R” groups are attached to biomolecules via a linker group and none of X, Y and Z is attached to a biomolecule via a linker group.  
     
     
         22 . Compound of  claim 19  wherein one of X, Y and Z is attached to a biomolecule via a linker group and none of the “R” groups are attached to biomolecules via a linker group.  
     
     
         23 . Compound of  claim 19  wherein a maximum of one “R” group attached to the carbon atoms of the macrocycle located between nitrogen atoms has a biomolecule attached via a linker group.  
     
     
         24 . Compound of  claim 19  wherein at least one of the “R” groups, in addition to the “R” groups which are attached to biomolecules via a linker group, are independently selected from the group consisting of alkyl, cycloalkyl, cycloalkylalkyl, aralkyl, alkaryl, aryl, heterocyclics and radicals attached to the α-carbon of α-amino acids, and the remaining “R” groups are independently selected from hydrogen, saturated, partially saturated or unsaturated cyclics or a nitrogen containing heterocycle.  
     
     
         25 . Compound of  claim 24  wherein at least two of the “R” groups, in addition to the “R” groups which are attached to biomolecules via a linker group, are independently selected from the group consisting of alkyl, cycloalkyl, cycloalkylalkyl, aralkyl, alkaryl, aryl, heterocyclics and radicals attached to the α-carbon of α-amino acids.  
     
     
         26 . Compound of  claim 24  wherein at least one of the “R” groups, in addition to the “R” groups which are attached to biomolecules, via a linker group, are alkyl and the remaining “R” groups are independently selected from hydrogen or saturated, partially saturated or unsaturated cyclics.  
     
     
         27 . Compound of  claim 19  wherein at least one of R 1  or R′ 1  and R 2  or R′ 2 , R 3  or R′ 3  and R 4  or R′ 4 , R 5  or R′ 5  and R 6  or R′ 6  R 7  or R′ 7  and R 8  or R′ 8  and R 9  or R′ 9  and R or R′ together with the carbon atoms to which they are attached represent a saturated, partially saturated or unsaturated cyclic having 3 to 20 carbon atoms and the remaining “R” groups in addition to the “R” groups which are attached to biomolecules via linker groups are independently selected from hydrogen, nitrogen containing heterocycles or alkyl groups.  
     
     
         28 . Compound of  claim 27  wherein at least two of R 1  or R′ 1  and R 2  or R′ 2 , R 3  or R′ 3  and R 4  or R′ 4 , R 5  or R′ 5  and R 6  or R′ 6 , R 7  or R′ 7  and R 8  or R′ 8 , and R 9  or R′ 9  and R or R′ together with the carbon atoms to which they are attached represent a saturated, partially saturated or unsaturated cyclic having 3 to 20 carbon atoms and the remaining “R” groups in addition to the “R” groups which are attached to biomolecules via linker groups are independently selected from hydrogen, nitrogen containing heterocycles or alkyl groups.  
     
     
         29 . Compound of  claim 27  wherein said saturated, partially saturated or unsaturated cyclic is cyclohexyl.  
     
     
         30 . Compound of  claim 29  wherein said remaining “R” groups in addition to the “R” groups which are attached to biomolecules via linker groups are independently selected from hydrogen or alkyl groups.  
     
     
         31 . Compound of  claim 19  wherein said R or R′ and R 1  or R′ 1 , R 2  or R 2 ′ and R 3  or R 3 ′, R 4  or R′ 4  and R 5  or R 5 ′, R 6  or R 6 ′ and R 7  or R 7 ′, and R 8  or R′ 8  and R 9  or R 9′  together with the carbon atoms to which they are attached are found to form a nitrogen containing heterocycle having 2 to 20 carbon atoms, and the remaining “R” groups in addition to the “R” groups which are attached to biomolecules via a linker group are independently selected from hydrogen, saturated, partially saturated or unsaturated cyclics or alkyl groups.  
     
     
         32 . Compound of  claim 19  wherein X, Y and Z are independently selected from the group consisting of halide, organic acid, nitrate and bicarbonate anions.