Metal complexes of hydroxyaryl containing aminocarboxylic acid chelating agents

Abstract

A physiologically acceptable, radiolabelled compound of general formula (I) consisting of a compound of the ##STR1## where k is an integer from 2 to 5l is an integer from 1 to 5X and Y are independently selected from phenyl, naphthyl, pyridine or quinoline radical, each having at position 2 one of --OH, --SH, --NH.sub.2 or --COOH and each of X and Y can be optionally substituted by one or more amino, halogen, hydroxy, mercapto, nitro, cyano, thiocyano, alkyl, alkoxy, halogenoalkyl, acyl, acylamino, acyloxy, carboxyl, alkoxycarbonyl, carbamoyl, pyridoylamino, N-carboxyalkyl-carbamoyl, sulpho, sulphamoyl, mono- or dialkylated or phenylated sulphamoyl which can also carry one or more substituents R', alkylsulphonyl, alkoxysulphonyl, or by an optionally hydroxy-containing phenylsulphonyl or phenoxysulphonyl; where R' is as defined for X and Y; or pharmaceutically acceptable salts thereof.

Description

The present invention relates to radiolabelled derivatives of ethylenediaminetetraacetic acid (EDTA) suitable for use as radiopharmaceutical agents. In particular, the present invention relates to EDTA derivatives, which when complexed with radionuclides may be used.as imaging agents for example to assess hepatobiliary function or in radiolabelling of monoclonal antibodies, proteins, peptides, oligonucleotides and the like for in vivo imaging or therapy.

U.S. Pat. No. 3,833,590 describes a class of EDTA derivatives which are useful for the control of metal-deficiency phenomena in biological systems. We have found that these derivatives are useful as radiopharmaceutical agents for example, for studying organ function or in radiolabelling of monoclonal antibodies and the disclosure of U.S. Pat. No. 3,833,590 is incorporated herein by reference.

Thus, according to one aspect, the present invention provides radiolabelled compounds of general formula (I) ##STR2## where k is an integer from 2 to 5

l is an integer from 1 to 5 X and Y are independently selected from phenyl, naphthyl, pyridine or quinoline radical, each having at position 2 one of --OH, --SH, --NH.sub.2 or --COOH and each of X and Y can be optionally substituted by one or more amino, halogen, hydroxy, mercapto, nitro, cyano, thiocyano, alkyl, alkoxy, halogenoalkyl, acyl, acylamino, acyloxy, carboxyl, alkoxycarbonyl, carbamoyl, pyridoylamino, N-carboxyalkyl-carbamoyl, sulpho, sulphamoyl, mono- or dialkylated or phenylated sulphamoyl which can also carry one or more substituents R', alkylsulphonyl, alkoxysulphonyl, or by an optionally hydroxy-containing phenylsulphonyl or phenoxysulphonyl; where R' is as defined for X and Y; or pharmaceutically acceptable salts thereof.

Preferably, X and Y are the same. The group --OH, --SH, --NH.sub.2, or --COOH is a donor functional group which is ortho to the position of attachment of the ring to the --NH of formula (I).

Compounds of formula (I) and salts thereof, are labelled by nuclides such as technetium-99m (.sup.99m Tc), copper-64(.sup.64 Cu) or copper-67(.sup.67 Cu) and subsequently administered intravenously into a subject for evaluating organ function, for example, the hepatobiliary system and for the purpose of therapy. The specificity for various organs may be achieved by the substitution of hydrophobic or hydrophilic groups on the ring structure at positions X and Y. Preferably, X and Y are each phenyl. For example, specificity for the liver may be achieved by the substitution of hydrophobic groups such as a chloride or a tertiary butyl group on the phenyl ring. The technetium complexes of some of the compounds of formula (I) have shown some specificity, high uptake and prompt excretion into the urine in rats and mice. The compound of formula (I) and salts thereof can also be labelled with radionuclides, such as .sup.67 Ga, .sup.111 In, .sup.57 Ni, .sup.57 Co, .sup.55 Co, .sup.166 Dy, .sup.166 Ho and other radionuclides from metals such as Y, Re, Fe, Au, Ag, Pb, Bi, Rh, Hg, Ti and Lanthanides such as Sm or Gd.

The radiolabelling of compounds of formula (I), and salts thereof can be accomplished by using procedures recognised in the art. The radiolabelling of the chelator with copper-67 can be achieved by adding copper in an aqueous acetate solution to a compound of formula (I) in an aqueous solution and incubating for 15 minutes at room temperature.

Alternatively, the radiolabelling of a chelator of formula (I) with technetium, for example, may be achieved by adding a reducing agent such as stannous chloride, to an aqueous solution of a compound of formula (I), followed by reaction with aqueous sodium pertechnetate solution (Na.sup.99m TcO.sub.4). The order of mixing these three components is believed not to be critical. However, preferably the reducing agent is added to the chelator of formula (I). The non-radioactive composition can be supplied to the radio-chemists, technicians, radio-pharmacists, doctors and the like for radiolabelling prior to use.

Technetium-99m in the form of an aqueous solution of sodium pertechnetate is readily obtainable from commercially available molybdenum-99/technetium-99m generators or alternatively, instant .sup.99m Tc may be used. .sup.64 Cu is commercially available from Australian Nuclear Science & Technology Organisation and .sup.67 Cu from the US Department of Energy, Brookhaven, USA.

The compounds of formula (I) are obtainable by the reaction of the anhydride of EDTA with one or two equivalents, as desired, of the corresponding amine. The reaction is performed in an inert-gas atmosphere and in the presence of solvent and diluents preferably inert to the reactants. Suitable solvent are aliphatic, aromatic or halogenated hydrocarbons such as benzene, toluene, xylenes, chlorobenzene, chloroform, methylene chloride, ethylene chloride; ethers and ethereal compounds such as dialkyl ether, ethylene glycol mono or -dialkyl ether, THF, dioxane; alkanols such as methanol, ethanol, n-propanol, isopropanol; ketones such as acetone, methyl ethyl ketone; nitriles such as acetonitrile or 2-methoxypropionitrile; N,N-dialkylated amides such as dimethylformamide; dimethylsulphoxide, tetramethylurea; as well as mixtures of these solvents with each other. If the amine or a salt thereof is soluble in water, then the reaction medium may be water at low temperature.

The compounds of formula (I) can be converted to pharmaceutically acceptable salts using art-recognised procedures.

In a second aspect, the present invention provides a diagnostic formulation suitable for labelling with .sup.99m Tc, .sup.188 Re or .sup.186 Re comprising a compound of formula (I) or a pharmaceutically acceptable salt thereof and a reducing agent in a pharmaceutically acceptable carrier.

In a third aspect, the present invention provides a formulation comprising a radiolabelled compound of formula (I) or a pharmaceutically acceptable salt thereof in a pharmaceutically acceptable carrier.

In a fourth aspect, the present invention provides a method of diagnosis or therapy comprising administering to a subject a radiolabelled compound of formula (I) or a pharmaceutically acceptable salt thereof.

We have further found that the nitro substituents on the X and Y groups of compounds of formula (I) may be further modified to form new compounds of formula (II). The synthesis of these new compounds has provided an opening to a new array of compounds, which have been found to be suitable for radiolabelling of monoclonal antibodies, receptor specific proteins, peptides, or oligonucleotides.

Radiolabelled antibodies are especially useful in medicine, for example, in locating specific tissue types and in the treatment of cell disorders. The labelled antibodies can be used to target metal ions to a specific tissue type, both in vitro and in vivo.

Thus, in a further aspect, the present invention provides novel compounds of formula (II), pharmaceutically acceptable salts and/or radiolabelled complexes thereof ##STR3## where k is an integer from 2 to 5

l is an integer from 1 to 5 R is independently selected from ##STR4## wherein Ar is an aryl or heteroaryl group; R.sup.1 is --NR.sup.2 R.sup.3 where R.sup.2 and R.sup.3 are independently selected from hydrogen, --(CH.sub.2).sub.p --NH.sub.2 ; --(CH.sub.2).sub.p --AR--(CH.sub.2).sub.m --NH.sub.2 ; --(CH.sub.2).sub.p --CO.sub.2 H; --(CH.sub.2).sub.p --AR--CO.sub.2 H; --(CH.sub.2 CH.sub.2 O).sub.n --CH.sub.2 CH.sub.2 NH.sub.2 ; --(CH.sub.2).sub.p --NCS, --(CH.sub.2).sub.p --AR--NCS; or --(CH.sub.2).sub.p NHCOR", --COR"; or R.sup.2 and R.sup.3 together with the nitrogen atom to which they are attached form an optionally substituted saturated or partially unsaturated ring optionally containing one or more heteroatoms O, S or N; or R.sup.1 is --NCS, --N.dbd.N or --C(.dbd.NH)--OCH.sub.3 ; n and m are independently an integer from 0 to 4 p is an integer from 1 to 4; R" is alkyl-L where L is halogen or other leaving group; 2,5-diketo-pyrrolinyl, --Het--CH.dbd.CH.sub.2 wherein Het is an optionally substituted 5 or 6 membered heterocyclic ring containing one or more heteroatoms O, N or S.

Preferably, aryl is phenyl, naphthyl, anthracenyl or the like and heteroaryl is preferably pyridine, quinoline, imidazolyl or the like. Het is preferably pyridine and Het--CH.dbd.CH.sub.2 is preferably, 2- or 4-vinylpyridine.

In yet a further aspect, the present invention provides a process of preparing compounds of formula (II), which comprises treating the corresponding nitro compound with a catalyst such as palladium/C in the presence of a reducing agent such as sodium borohydride and as required, further derivatising the amino group following procedures known in the art.

Compounds of formula (II) where R.sup.1 is mono- or di-substituted amino and where the substituent is substituted alkyl, are readily prepared by treating the amino compound with the appropriate halo-substituted alkyl. For example, compound of formula (II) where R.sup.1 is --NH--CH.sub.2 --CH.sub.2 --NH.sub.2 can be prepared by treating a compound of formula (II) where R.sup.1 is NH.sub.2 with BrCH.sub.2 CH.sub.2 NH.sub.2 in the presence of NaHCO.sub.3 or the like.

Compounds of formula (II) where R.sup.1 is --NCS is prepared by reacting the amino compound with thiophosgene (see WO87/12631), Kozak et al., Cancer Res. 49, 2639 (1989). Substituted acid halide compounds are produced by reacting a compound of formula (II) where R.sup.1 is NH.sub.2 with BrCH.sub.2 COBr at 4.degree. C. according to the procedure of C J Mathias et al., Bioconjugate Chem., 1, 204 (1990). Compounds with an electrophilic moiety can also be prepared by methods known in the art, such as in ACC Chem. Res. 17 202-209 (1984). Compounds with active esters R--C(O)--X can be formed by the procedures of Bodanszky M, The Peptide. Analysis. Synthesis. Biology, Ed. E. Gross and J Meienhofer, Vol 1. pp105-196, Academic Press, Inc., Orlando, FL. (1979) and Bodanszky M, Principles of Peptide Synthesis, pp9-58, Springerverlag, New York, (1984). 2,5-diketo-pyrrolinyl, --Het--CH.dbd.CH.sub.2 or 2,4-vinylpyridine compounds can be prepared according to the procedure of Morphy et al., J. Chem. Soc., Chem. Commun., 156, (1988). Compounds containing maleimides and succinimide may be prepared by procedures outlined in Y Arano et al., Bioconjugate Chem., 2, 71, (1991) and S Garg et al., Bioconjugate Chem., 2, 50, (1991). Other standard procedures are to be found in Modern Synthetic Reactions, H O House, 2nd Edition, Benjamin, Inc. Philippines, 1972.

The R.sup.1 group may be termed the reactive group which provides the point of attachment for a protein such as a monoclonal antibody either directly, ie where R.sup.1 is NH.sub.2, or through a linker, ie other R.sup.1 groups defined above. The isothiocyanate and the halogen functionalised derivatives may be directly linked to a protein (eg. antibody) thiol or amino group. In a structural comparison, it can be seen that R.sup.1 is equivalent to a substituent on the X or Y group of compounds of formula (I).

Preferably, the --OH group is ortho to the position of attachment of the phenyl ring to the --NH of formula (II). R.sup.1 can be on any other position on the ring.

Compounds of formula (II) provide a method of attachment of radionuclide metal ions such as In(III), Gd(III), Ga(III), Fe(III), TcO.sub.4.sup.3-, Cu(II), Ti(IV) and other radionuclides from the Lanthanides, Rhenium, Samarium, Holmium, Yttrium and the like to monoclonal antibodies, receptor specific proteins, peptides or oligonucleotides for in vivo imaging and therapy.

Preferred use of the compounds would be to radiolabel monoclonal antibodies specific for colon, ovarian, lymphoma, breast and/or bladder cancer, with beta emitter radionuclides of metals such as Sm, Ho, Re, Sc, Cu, Pd, Pb, Rh and Y for therapy of above mentioned cancer. A further preferred use is in the radiolabelling of a monoclonal antibody specific for metastasis of colon cancer for diagnosis and therapy.

It is generally preferable to couple the compounds of formula (II) to other molecules such as proteins, peptides, antibodies or carbohydrates to form conjugate compounds for use as radiopharmaceutical agents or cytotoxic agents.

Compounds of formula (II) may be coupled through any thiol, amino, carboxyl, hydroxyl, aldehyde, aromatic or heteroaromatic group present in the protein, peptide, carbohydrate or oligonucleotide.

In particular, the present invention provides a conjugate compound which comprises a compound of formula (II), a radiolabelled complex and/or salt thereof, coupled to an antibody, protein, peptide, carbohydrate or oligonucleotide.

The conjugate compound according to the invention may contain more than one molecule of a compound of formula (II) coupled to any one protein, peptide or carbohydrate molecule. The antibody in the conjugates may be a complete antibody molecule or a fragment thereof or an analogue of either of these, provided that the antibody comprises a specific binding region. The antibody may be humanized monoclonal or a fragment thereof. The antibody may also be a recombinant antibody. The antibody may be specific for any number of antigenic determinants, but is preferably specific for one antigenic determinant.

In another aspect, the present invention provides a pharmaceutical formulation comprising a compound of formula (II), a radiolabelled complex and/or salt thereof in a pharmaceutically acceptable carrier.

In a further aspect, the present invention provides a method of diagnosis or therapy in a subject comprising administering to the subject a compound of formula (II) or a radiolabelled complex and/or salt thereof.

Radiolabelling of proteinaceous materials using bifunctional chelators of the present invention can be conducted in two ways, namely:

(a) prelabelling of chelator followed by conjugation of resultant radiocomplex to proteinaceous material, or (b) conjugating chelator to proteinaceous material for subsequent radiolabelling.

Other suitable applications are:

(a) PET imaging

When chelators of formula (I) or (II) are labelled with .sup.64 Cu (Positron 0.278 MeV; half-life 12.70 hours), or .sup.62 Cu (Positron 0.93 MeV; half-life 9.8 minutes), such radiocomplexes may be useful for quantitative dynamic flow measurements of blood flow and bile flow in the hepatobiliary system. Alternatively the chelators may be labelled with Ti-45, a positron emitter (1.04 MeV) with a half life of 3.08 hours.

(b) Radiolabelling monoclonal antibodies with .sup.67 Cu, (Beta and Gamma emitters) for combined radioimmunoscintography (RIS) and radioimmunotherapy (RIT)

(c) Auger emitting agents where a chelator is coupled to an intercalator and labelled with auger emitting isotope such as Fe-59

(d) two step pretargeted radioimmunotherapy

A monoclonal antibody with one or more biotin molecules or marker molecules attached is injected into a patient. Once the antibody has cleared from the system and localized to the tumour, a second injection is administered. This second injection involves the radiolabelled chelator attached to a smaller molecule such as avidin or streptavidin which recognises the biotin or marker on the targeted antibody.

Alternatively, the second injection may be avidin or streptavidin and when cleared from the system, the radiolabelled chelator attached to biotin is administered. Both procedures provide amplification of the target site and reduce exposure to normal tissue.

Another two step procedure involves the administration of an antibody-DNA conjugate or antibody-oligonucleotide conjugate followed by targeting with a radiolabelled complementary DNA or complementary oligonucleotide. This procedure also provides amplification of the target site and reduces exposure to normal tissue.

(e) Radiotherapy of Liver metastasis

The chelators compound 2 and compound 4 specifically traverse the hepatobiliary system (see data in Tables 2 and 4, Biodistribution of .sup.99m Tc complex of compound 2 and 4 respectively), and empty into the gastrointestinal tract. When chelators, such as compound 2 and compound 4, are labelled with a therapeutic radionuclide such as .sup.67 Cu, they would be useful for treatment of metastasis of the liver.

(f) Magnetic Resonance Imaging (MRI) agents

We envisage the use of these compounds as MRI agents where complexes formed with any paramagnetic metal ion such as Fe (III) may be used as a contrast agent to enhance images. Also complexes such as these may be attached to a pharmaceutically acceptable carrier for the same purpose. We also envisage the industrial use of these compounds for attachment to solid surfaces such as polymers, electrode surfaces etc. for use in the concentration of metal ions, purification of water or the like.

The present invention also encompasses mixed combinations of radioactive labels on a given monoclonal antibody or a mixture of monoclonals and metal radionuclides attached to one or more different ligands. These would be useful for cancer therapy.

Specific embodiments of the present invention are illustrated by the following examples. It will be understood, however, that the invention is not confined to specific limitations set forth in the individual examples and drawings.

A BRIEF DESCRIPTION OF THE DRAWINGS

FIG. 1A is a graph showing serum stability of compound (1) with Tc-99m at time=0 hr.

FIG. 2B is a graph showing serum stability of compound (1) with Tc-99m at time=1 hr.

FIG. 2A is a graph showing serum stability of compound (1) with Cu-67 at time=0 hr.

FIG. 2B is a graph showing serum stability of compound (1) with Cu-67 at time=1 hr.

FIG. 2C is a graph showing serum stability of compound (1) with Cu-67 at time=2 hr.

FIG. 2D is a graph showing serum stability of compound (1) with Cu-67 at time=4 hr.

FIG. 2E is a graph showing serum stability of compound (1) with Cu-67 at time=24 hr.

FIG. 3 is a graph showing the effect of protein concentration on cross-linking when radiolabelling with Cue.sup.54/57 compound (5).

FIG. 4 is a graph showing the rate of complexation of .sup.67 Ga(compound 1) at T=21.degree. C.

FIG. 5 is a graph showing the rate of complexation of .sup.67 Ga(compound 1) at T=30.degree. C.

FIG. 6 is a graph showing the rate of complexation of .sup.67 Ga(compound 1) at T=36.degree. C.

FIG. 7 is a graph showing the rate of complexation of .sup.67 Ga(compound 2) at T=21.degree. C.

FIG. 8 is a graph showing the rate of complexation of .sup.67 Ga(compound 2) at T=30.degree. C.

FIG. 9 is a graph showing the rate of complexation of .sup.67 Ga(compound 2) at T=37.degree. C.

FIG. 10 is a graph showing the rate of complexation of .sup.67 Ga(compound 4) at T=21.degree. C.

FIG. 11 is a graph showing the rate of complexation of .sup.67 Ga(compound 4) at T=25.degree. C.

FIG. 12 is a graph showing the rate of complexation of .sup.67 Ga(compound 4) at T=30.degree. C.

FIG. 13 is a graph showing the rate of complexation of .sup.67 Ga(compound 4) at T=37.degree. C.

FIG. 14 is a graph showing the rate of complexation of .sup.67 Ga(compound 5) at T=21.degree. C.

FIG. 15 is a graph showing the rate of complexation of .sup.67 Ga(compound 5) at T=35.degree. C.

FIG. 16 is a graph showing the rate of complexation of .sup.67 Ga(compound 6) at T=21.degree. C.

FIG. 17 is a graph showing the rate of complexation of .sup.67 Ga(compound 6) at T=27.degree. C.

FIG. 18 is a graph showing the rate of complexation of .sup.67 Ga(compound 6) at T=32.degree. C.

FIG. 19 is a graph showing serum stability .sup.67 Ga(compound 1) at t=0.

FIG. 20 is a graph showing serum stability .sup.67 Ga(compound 1) t=6 days

FIG. 21 is a graph showing serum stability .sup.67 Ga(compound 2) t=0.

FIG. 22 is a graph showing serum stability .sup.67 Ga(2 compound) t=6 days

FIG. 23 is a graph showing serum stability .sup.67 Ga(compound 6) t=0

FIG. 24 is a graph showing serum stability .sup.67 Ga(compound 6) t=22 h

FIG. 25 is a graph showing serum stability .sup.67 Ga(compound 6) t=46 h

FIG. 26 is a graph showing biodistribution of .sup.67 Ga(compound 1)

FIG. 27 is a graph showing biodistribution of .sup.67 Ga(compound 2)

FIG. 28 is a graph showing biodistribution of .sup.67 Ga(compound 4)

FIG. 29 is a graph showing biodistribution of .sup.67 Ga(compound 5)

FIG. 30 is a graph showing biodistribution of .sup.67 Ga(compound 6)

FIG. 31 is a graph showing serum stability elution profile of .sup.166 Dy/.sup.166 Ho(compound 1) at 46 hours.

EXPERIMENTAL

A series of derivatives incorporating carboxyl and phenolate donor groups (1-6) were synthesised by literature method and their radiolabelling characteristic and biological behaviour evaluated. Initial work involved an investigation of radiochemical properties of the .sup.99m Tc complexes of the ligands and their biodistribution in rats and mice (see Tables 1-6 and 8-12).

Compound No. 3

To a solution of 4-nitro-2-amino-phenol (0.18 g) in acetonitrile (100 ml) was added EDTA anhydride (5.05 g). The reaction mixture was stirred vigorously with reflux under N.sub.2 gas overnight. The solid which formed was filtered off while the solution was still warm. The yellow solid was then washed with copious amount of methanol and finally acetone. Yield: 8.92, 81%.

All compounds listed below were prepared in a similar manner. Yields and all spectral data for some compounds are presented in Table A.

______________________________________Compound No. X and Y Formula______________________________________ ##STR5## I2 ##STR6## I3 ##STR7## I4 ##STR8## I5 ##STR9## II6 ##STR10## I______________________________________ TABLE A__________________________________________________________________________Cpd Mpt.sup.b FTIR.sup.c NMR.sup.d(yield %) Ms.sup.a (.degree.C.) (cm.sup.-1) (ppm)__________________________________________________________________________No. 1 475 204-205 NCCO, 1708.2 8.02, s, 2H, OH; 6.91-6.72, m, 6H; Ar; 3.46, 3.38, s, 4H, 4H,(95%) COOH, 1635.8 CH.sub.2 CONH, CH.sub.2 COOH; 2.86, s, 4H, NCH.sub.2 CH.sub.2 ; OH, 3314.1No. 2 543 222-224 NHCO, 1716.9 9.63, s, 2H, OH; 8.17, 6.01, 6.81, d, 6H, Ar; 0.45, 3.39, s, 4H,(79%) COOH, 1659.0 4H, CH.sub.2, CONH, CH.sub.2 COOH; 2.83, s, 4H, NCH.sub.2 CH.sub.2 ; OH, 3331.6No. 3 564 222-226 NHCO, 1728.4 9.88, s, 2H, OH; 9.06, 7.89, 6.94, d, 2H, 2H, Ar; 3.56, 3.52, s,(81%) COOH, 1693.9 4H, 4H, CH.sub.2 CONH, CH.sub.2 COOH; 2.83, s, 4H, NCH.sub.2 CH.sub.2 ; OH, 3299.3No. 4 587 .degree. NHCO, 1708.9 9.59, s, 2H, OH; 8.11, 6.90, 6.75, d, 6H, Ar; 3.42, 3.38, s, 4H,(>85%) COOH, 1658.8 4H, CH.sub.2 CONH, CH.sub.2 COOH; 2.83, s, 4H, NCH.sub.2 ; 1.21, s, OH, 3318.9 18H, tBu;No. 5 509 ' NHCO, 1740.1 9.33, s, 2H, OH; 8.41-8.28, m, 6H, Ar; 5.42, 5.35, s, 4H, 4H,Method A COOH, 1683.3 CH.sub.2 CONH, CH.sub.2 COOH; 3.80, s, 4H, NCH.sub.2 CH.sub.2 ;(95%) OH, 3300 brNo. 5 504 ' NHCO, 1734.0 .sup.e 7.52, s, 2H, 6.90-6.81, m, 4H, Ar; 3.92, 3.76, s, 4H, 4H,Method B COOH, 1684.0 CH.sub.2 CONH, CH.sub.2 COOH; 3.28, s, 4H, NCH.sub.2 CH.sub.2 ;(95%) OH, 2998 br__________________________________________________________________________ .sup.a Positive FAB mass spectra were determined of Jeol DX300 mass spectrometer. MS: parent Ion (FAB): M/C, LH.sup.+. .sup.b All melting points are uncorrected. .sup.c Diffuse reflectance FT Infrared spectra were recorded on BioRad Giglab FTS60 in 1% Kbr. .sup.d The .sup.1 H NMR spectra were determined in d.sub.6 DMSO at 298'K, on a Jeol Gx400 MHz spectrometer. .sup.e The .sup.1 H NMR spectra were determined in D.sub.21 O at 298'K, o a Jeol Gx400 MHz spectrometer. .degree.Compound hygroscopic. 'Decomposes. Preparation of New Compound No. 5

Method A

To Palladium/C catalyst (50 mg) in water (10 ml) was added sodium borohydride (0.24 g) in water (10 ml) under nitrogen gas. To this mixture was then added compound 3 (1.0 g) in ethanol/sodium hydroxide solution (10 ml/1 ml, 8% NaOH) slowly. The mixture was left to stir at room temperature for a further 20 minutes or till the solution becomes clear. To this mixture is added concentrated hydrochloric acid dropwise till all excess sodium borohydride is quenched. Palladium catalyst is filtered off and the filtrate is reduced under vacuum. The pale pink powder is dissolved in a minimum of methanol and any salt remaining filtered off. Yield: 0.85 g; 95%.

Method B

To palladium on activated charcoal (50 mg) in distilled water (2 cm.sup.3) was added sodium borohydride (0.16 g) under nitrogen. Ethanol (.about.8 cm.sup.3) was added to the reaction mixture, then ligand (3) (1.0 g) in concentrated ammonia (10 cm.sup.3 :28% ammonia). The mixture was left to stir at room temperature for a further 30 minutes or until the solution became clear in colour. Concentrated hydrochloric acid was added dropwise to the mixture to destroy any unreacted sodium borohydride or until gases ceased to evolve. The final pH of the solution was .about.6.5.

The palladium catalyst was removed by filtration and the filtrate reduced under vacuum. Water was azeotroped off using copious amounts of ethanol and acetone. The purple/brown residue was suspended in a minimum amount of ethanol (.about.20 cm.sup.3) and any undissolved salt removed by filtration. Yield=0.85 g; (95%). Final product was characterised by .sup.1 H NMR, FT-IR and electrospray mass-spectrometry and melting point. Data are tabulated in Table A.

Radiolabelling of Chelators with .sup.99m Tc

A typical method of radiolabelling of the chelator was achieved by dissolving the chelator in a basic solution of saline. The pH was then adjusted to approx. 6 with HCl. Excess stannous chloride was added to the resultant solution followed by one equivalent volume of sodium .sup.99m TcO.sub.4 (10-50 mCi/ml). Final concentration of the chelator was approx. 1 mg/ml. The efficacy of labelling was determined by ITLC-SG. For each product only one main radiochemical species was observed. Each complex was diluted to 15 .mu.Ci/100 .mu.l with saline. In vitro saline stability studies showed the complexes were stable for up to 6 hours. Serum stability studies indicated that no more than 15 per cent of the technetium is lost from the chelator (1) during the first hour at 37.degree. C. (See FIGS. 1A and 1B).

Biodistribution studies in rats show that the .sup.99m Tc complexes are extracted rapidly and efficiently from the blood by the kidney and the liver.

Selected data from biodistribution results of technetium-99m complexes of No. 1, No. 2 and No 4 in the liver, kidney and blood at 3 minutes is tabulated below.

______________________________________Compound Liver Kidney Blood______________________________________No. 1 1.54 4.12 1.16No. 2 5.93 8.90 1.32No. 4 7.89 4.54 1.71______________________________________

The structural variation in this selection of derivatives is the substitution of a chloride atom in No. 2 for a hydrogen atom on both aromatic rings. For No. 3, the substitution of a tertiary butyl group on each aromatic ring. The substitution of these groups make the parent compound more hydrophobic hence there becomes a preference for the liver uptake as opposed to the kidney.

Effect of pH on Complexation of .sup.99 mTC with ligand (5)

Radiolabelling with .sup.99m Tc was investigated over a range of pHs (3.0-10.0) (described in Table B). To ligand (5) (1.0 ml:1 mg/ml) dissolved in appropriate buffer was added an excess of stannous chloride (20 .mu.l:2 mg/ml) in 0.1 N HCl. To this mixture is added an appropriate activity of .sup.99m TcO.sub.4 (20 .mu.l) eluted from a technetium generator.

All reactions were carried out at room temperature.

Percent complexation was monitored by ITLC-SG using two solvent systems. System 1: cyclohexane/acetone 1:2 showed free .sup.99m TcO.sub.4.sup.- migrated with the solvent front and any colloid or complex formed remained at the origin. System 2: saline showed complex and free .sup.99m TcO.sub.4 migrate with the solvent front and colloid remained at the origin. Results are illustrated in Table C.

TABLE B______________________________________Buffers and range of pH used in .sup.99m Tc and .sup.64/67 Curadiolabelling of (5)pH range Buffer (0.5 M)______________________________________3.0-4.0 acetate5.0-7.0 phosphate8.0-9.0 bicarbonate10.0 carbonate______________________________________ TABLE C______________________________________Percent .sup.99m Tc complexed by ligand (5) versesreaction timeReaction TimepH 1 min 3 min 5 min 10 min 15 min 30 min 1 h 2 h______________________________________3 89 93 90 91 90 92 92 944 94 97 92 94 96 91 100 1005 84 83 91 83 81 71 85 886 80 80 83 80 82 90 85 917 73 83 81 74 87 80 83 528 76 88 81 83 87 85 77 819 90 100 94 86 100 83 85 7410 15 16 11 13 9 ND 8 8______________________________________

This study shows that complexation for (5) is rapid and optimum at pH 3.0-4.0. The final product formed is stable for upto 2 hours throughout pH range 3.0-9.0. However complexation is poor at pH 10.0 and what product formed is unstable.

Radiolabelling of Chelators with .sup.64 Cu and .sup.67 Cu

A typical procedure involved the dissolving Cu-67 20 .mu.l of original stock in an acetate buffer (0.2 M; pH 4.5) to a total volume of 500 .mu.l. To this solution was added 500 .mu.l of the ligand (20 mg/ml) solution in acetate buffer. The mixture was then left to react at 37.degree. C. for 20 minutes. ITLC confirmed the labelling was greater than >95%.

Serum Stability Studies of Copper-67 Complexes

The serum stability was determined for selected derivatives and typical procedure is as follows.

The ligand (1) was labelled with Cu-67 at a ligand:metal ratio of 4:1. ITLC showed the efficiency of labelling to be >95%. Serum stability studies demonstrated a loss of 4% over the initial 4 hours and no further loss after 24 hours. See FIGS. 2A, 2B, 2C, 2D and 2E.

Biodistribution of Copper-67 Complex of (5)

The biodistribution of the copper-67 complex of ligand (5) was evaluated in balb/c mice (5 animals per time point) at 1, 3, 5, 10, 30 minute time intervals. The data are presented in Table 7.

The results show that the Cu-67 complex of (5) clears from the kidneys, blood and urine over the 30 minute period. However there appears to be an active uptake of the complex in the liver over the 30 minute period.

Effect of pH on radiolabelling liggnd (5) with .sup.64/67 Cu

Radiolabelling with .sup.64/67 Cu was investigated with ligand (5) over a range of pHs (see Table B). Conditions employed are similar to that reported above for where the .sup.64/67 Cu in appropriate buffer was added directly to a buffered solution of ligand (5). Rate and percent of complexation was monitored over time by ITLC-SG. Mobile phase employed was methanol:water:ammonia (9:1: trace amounts). The copper complex moves with the solvent front while the free copper is retained at the origin. Complexation was complete within the first minute for all pHs (pH 3.0-10.0) and maintained for upto 2 hours.

Radiolabelling of Protein

Radiolabelling of protein may be achieved by two methods (a) First the chelator may be radiolabelled by a procedure outlined previously. The radiolabelling of the proteinaceous material may be accomplished using a cross-conjugation linker such as EDC (CH.sub.3 CH.sub.2 --N.dbd.C.dbd.N--(CH.sub.3).sub.3 --NH.sup.+ --(CH.sub.3).sub.2 --HCl or the like. For example, to IgG (such as B72.3) in phosphate buffered saline is added the radiolabelled ligand at a molar ratio of 20:1 and EDC in a 1000:1 molar ratio. The mixture is then left to react at 37.degree. C. for 11 hour. The unreacted reagents are removed by size exclusion ultra filtration membrane.

Radiolabelling of Protein with .sup.67 Cu

The radiolabelling of human serum albumin (HSA, Sigma Chemicals) and streptavidin (Sigma Chemicals) and IgG was achieved at values ranging from 0.12-0.16.mu.Ci/.mu.g. The purified radiolabelled IgG was analyzed on HPLC size-exclusion column and fractions collected. Greater than 80% was associated with the main protein peak. No evidence of crosslinking was observed under the labelling conditions used, ie similarly, the chelators can be labelled with a range of radionuclides such as .sup.62 Cu, .sup.166 Ho, .sup.153 Sm, .sup.188 Re, .sup.186 Re and .sup.45 Ti.

Typical reaction for radiolabellinc B72.3 antibody with .sup.64/67 CU

To ligand (5) dissolved in 0.2 M sodium acetate buffer (pH 7.0) was added .sup.64/67 Cu in acetate buffer. Greater than >99w complexation was confirmed by ITLC-SG. Antibody (5-15 mg/ml) was then reacted with the solution of complex (5) at a 20:1 ligand to antibody ratio in the presence of 1000:1 EDC at 37.degree. C. A range of antibody concentrations were investigated to determine the effect on cross-linking of protein. Each product was purified by size-exclusion chromatography and then analysed for cross-linking using HPLC Bio-Sil SEC-250 column (eluent PBS: 0.7 ml/min). The results are illustrated in FIG. 3. The data shows that as the protein concentration was increased the amount of cross-linking also increased.

(b) The second method involves the conjugation of the chelator to the antibody or proteinaceous material as described but using the unlabelled ligand and prepared in a form suitable for subsequent labelling with a suitable radionuclide. This is the preferred form for preparation of the proteinaceous material in a kit form, suitable for manufacturing.

Ga-67 citrate (t.sub.1/2 =78.25 h, y=184.6 keV (19.7%)) is commonly used in the diagnosis of lymphoma. While Ga-67 citrate shows relatively fast and selective uptake at the disease sites, it clears slowly from the blood, through the liver, accumulating in the bowel. Its long biological half-life necessitates repeated scanning at delayed intervals (upto 72 h postinjection), resulting in repeated visits of the patient and increased demands on physician time.

In an attempt to address these short comings, .sup.67 Ga-complexes of a series of hydroxyaryl EDTA derivatives (1)-(6) have been synthesized and their mode complexation and biological behaviour investigated.

Radiolabelling (1)-(6) with .sup.67 Ga

The series of derivatives (1) to (6) were radiolabelled with .sup.67 Ga. The kinetics of radiolabelling were monitored at various pHs and temperatures. The optimum pH for complexation was .about.12. The data are represented in FIGS. 4-18.

A typical method for radiolabelling with .sup.67 Ga

Dissolve the ligand (1) �10 mg! in 1 M NaOH (200 ul) and then adjust the volume to 1 mL with 0.2 M NaOAc (pH 7.0). The final pH of solution should be .about.13. Dissolve the .sup.67 Ga citrate (purchased from Australian Radioisotopes, Australia) with 0.2 M NaOAc (pH 7.0) buffer. Mix equal volumes of the ligand and .sup.67 Ga NaOac solutions and monitor complexation by ITLC-SG (mobile phase ethanol:water 9:1). R.sub.f for .sup.67 Ga complex migrate with solvent front and any colloid or uncomplexed .sup.67 Ga remained at the origin.

Typical preparation of .sup.67 Ga complex for serum stability studies and biodistributions.

To ligand (50 .mu.L, 10 mg/mL, 0.2 M NaOAc, pH 12) was added .sup.67 Ga citrate (18.5 MBq) and the resultant mixture diluted to 1 mL (0.2 M NaOAc, pH 7). The pH was adjusted to 12 with 1 M NaOH (20 .mu.L) and heated at 40.degree. C. for 40 min. The final volume was adjusted (to 5 mL) with saline and sterilised by filtering through a 0.22 .mu.m filter.

Each animal was injected via tail vein with .sup.67 Ga complex (100 .mu.L, .about.3.7.times.10.sup.5 Bq). Animals were sacrificed at various time intervals (5 animals per time point/6 time points). Data are illustrated in FIG. 26-30 and in Tables 13-17.

Serum stability of the selected .sup.67 Ga-complexes.

The .sup.67 Ga complexes were prepared as described, then diluted in human plasma (5 mL).

The mixtures were incubated at 37.degree. C. for upto six days. Aliquots were sampled at regular intervals and the protein separated from the complexes using size-exclusion chromatography (sephadex-G25; eluent: phosphate buffered saline). Fractions (1.5 mL) were collected and monitored for TV absorbance and radioactivity. Profiles show no radioactivity associated with the protein indicating .sup.67 Ga complexes are stable under conditions described. See FIG. 19-25.

Biodistribution studies of .sup.67 Ga complexes

Each .sup.67 Ga-ligand complex has been evaluated in balb/c mice. Studies show that the complex route of excretion may be directed by the substituents on the phenolic groups of the ligand. (see FIG. 26-30).

The present study shows that complexation of the hydroxyaryl ligands is rapid (at pH>12) and the .sup.67 Ga complexes are stable. Data show that substituents on the phenolic rings of these ligands can effect the mode of excretion of the .sup.67 Ga complexes of the hydroxyaryl-EDTA derivatives. High yields in synthesis of the hydroxyaryl-EDTA derivatives and rapid complexation make these ligands attractive for "kit formulation" and indicate that they may have a role as imaging agents in nuclear medicine.

Complexation of these ligands have also been investigated with .sup.57 Ni, .sup.57 Co and .sup.55 Co.

Typical preparation involves dissolving the ligand (10 mg) in 0.1 N NaOH and then adjusting the pH to 6.5. The final mixture is then diluted with acetate buffer (0.2 N NaAc; pH 5.0). The radioisotope dissolved in acetate buffer is then added to the mixture. Complexation is confirmed by ITLC-SG using methanol: water: ammonia (9:1:trace). In each case the complexes move with the solvent front while free radioisotope remains at the origin. Under these conditions complexation was complete in less than 1 h.

Complexation and stability of (1). with .sup.166 Dy and .sup.166 Ho

Radiolabelling with of (1) with .sup.166 Dy and .sup.166 Ho and the serum stability of the complex formed has been investigated. A typical procedure involves dissolving (1) (2.6 10.sup.-4 M) in 1.0 M NaOH (150 .mu.l) and then adding saline (669 .mu.l). Then equal volume of ligand mixture and the .sup.166 Dy/.sup.166 Ho in 0.2 M NaAc (pH to 4.5) are mixed. The complexation is monitored by ITLC-SG. Mobile phase acetonitrile: water:4:1 shows >99% complexation is complete within 40 mins at RT.

The radiolabelling (1) is then incubated in human serum at 37.degree. C. At various time intervals the aliquots are separated and monitored for UV absorbance and radioactivity. The data illustrated in FIG. 31 show the complex is stable for >46 hrs.

Formulations of the radiolabelled compounds of formula (I) or (II) are prepared and administered in according to standard techniques. A pharmaceutical formulation of the present invention comprises the active compound together with one or more pharmaceutically acceptable carriers and optionally any other therapeutic ingredient. The formulation may conveniently be prepared in unit dosage form and may be prepared according to conventional pharmaceutical techniques. Additionally, the formulation may comprise one or more accessory ingredients such as diluents or buffers. The effective amount of the active compound will vary both with the route of administration, the condition under treatment and host undergoing treatment.

TABLE 1__________________________________________________________________________BIODISTRIBUTION OF .sup.99m TC COMPLEX OF NO. 1.sup.aMEAN SD MEAN SD MEAN SD MEAN SD MEAN SD30 SEC 1 MIN 3 MIN 5 MIN 10 MIN__________________________________________________________________________LIVER 1.73 0.21 2.01 0.15 1.54 0.10 1.34 0.15 0.76 0.15SPLEEN 0.53 0.04 0.42 0.04 0.26 0.03 0.20 0.03 0.15 0.01KIDNEYS 6.73 0.45 6.55 0.50 4.12 0.62 2.87 0.21 2.72 0.34MUSCLE 0.28 0.02 0.27 0.03 0.26 0.07 0.27 0.13 0.15 0.01SKIN 0.37 0.03 0.43 0.05 0.49 0.04 0.53 0.06 0.40 0.10BONE 0.31 0.03 0.32 0.03 0.26 0.02 0.25 0.07 0.19 0.02LUNGS 1.15 0.10 1.13 0.11 0.80 0.05 0.54 0.03 0.46 0.03HEART 0.75 0.04 0.67 0.09 0.46 0.06 0.33 0.03 0.27 0.03BLOOD 1.93 0.09 1.71 0.14 1.16 0.14 0.83 0.05 0.71 0.04STOMACH 0.14 0.08 0.28 0.14 0.32 0.28 0.11 0.01 0.12 0.03TOT GIT 0.27 0.02 0.34 0.03 0.59 0.19 0.79 0.08 1.22 0.16TAIL 0.68 0.03 0.56 0.05 0.80 0.32 0.73 0.04 0.83 0.10BRAIN 0.14 0.30 0.09 0.03 0.06 0.02 0.05 0.01 0.04 0.01THYROID 1.42 0.14 1.15 0.24 1.17 1.12 0.59 0.09 0.40 0.07EYES 0.19 0.02 0.17 0.01 0.18 0.02 0.17 0.01 0.13 0.02PANCREAS 0.49 0.08 0.51 0.11 0.44 0.19 0.44 0.18 0.30 0.02THYMUS 0.38 0.04 0.35 0.04 0.27 0.03 0.19 0.03 0.14 0.01__________________________________________________________________________ .sup.a Australian Albino Wistar Rats, % Injected dose per gram (X .+-. SD). Average of 5 animals per time point. TABLE 2__________________________________________________________________________BIODISTRIBUTION OF .sup.99m TC COMPLEX OF NO. 2.sup.aMEAN SD MEAN SD MEAN SD MEAN SD MEAN SD30 SEC 1 MIN 3 MIN 5 MIN 10 MIN__________________________________________________________________________LIVER 4.61 0.69 6.28 1.07 5.93 0.73 3.93 0.66 1.89 0.36SPLEEN 0.62 0.04 0.58 0.03 0.33 0.03 0.25 0.04 0.16 0.03KIDNEYS 6.42 0.45 8.37 0.63 8.90 0.73 7.26 2.45 3.35 0.54MUSCLE 0.22 0.03 0.24 0.01 0.20 0.03 0.16 0.02 0.11 0.02SKIN 0.29 0.02 0.45 0.07 0.42 0.02 0.35 0.02 0.33 0.05BONE 0.34 0.03 0.37 0.03 0.25 0.03 0.19 0.02 0.15 0.03LUNGS 1.20 0.09 1.17 0.14 0.79 0.06 0.59 0.02 0.48 0.09HEART 0.87 0.11 0.93 0.07 0.46 0.02 0.36 0.03 0.28 0.05BLOOD 2.79 0.33 2.77 0.20 1.32 0.09 0.94 0.08 0.77 0.16STOMACH 0.41 0.10 0.68 0.46 2.08 3.10 0.98 0.94 0.90 1.51TOT GIT 0.48 0.12 0.58 0.08 1.56 0.25 2.27 0.19 3.18 0.71TAIL 0.81 0.22 0.66 0.09 0.64 0.07 1.16 0.78 0.55 0.08BRAIN 0.14 0.01 0.09 0.01 0.06 0.01 0.04 0.01 0.03 0.01THYROID 1.41 0.36 1.71 0.66 0.81 0.18 0.58 0.14 0.44 0.12EYES 0.11 0.01 0.15 0.03 0.12 0.02 0.10 0.02 0.10 0.02PANCREAS 0.49 0.06 0.66 0.16 0.44 0.17 0.52 0.34 0.27 0.04THYMUS 0.39 0.05 0.37 0.08 0.27 0.04 0.17 0.02 0.15 0.03__________________________________________________________________________ .sup.a Australian Albino Wistar Rats, % Injected dose per gram (X .+-. SD). Average of 5 animals per time point. TABLE 3__________________________________________________________________________BIODISTRIBUTION OF .sup.99m TC COMPLEX OF NO. 3.sup.aMEAN SD MEAN SD MEAN SD MEAN SD MEAN SD30 SEC 1 MIN 3 MIN 5 MIN 10 MIN__________________________________________________________________________LIVER 1.43 0.03 1.61 0.18 1.61 0.19 1.24 0.20 0.92 0.16SPLEEN 0.49 0.06 0.42 0.05 0.29 0.03 0.23 0.03 0.17 0.01KIDNEYS 3.92 0.22 4.63 0.64 4.27 1.12 3.00 0.31 2.10 0.21MUSCLE 0.26 0.02 0.24 0.03 0.23 0.04 0.21 0.02 0.22 0.06SKIN 0.49 0.06 0.43 0.07 0.58 0.11 0.64 0.08 0.63 0.04BONE 0.30 0.03 0.29 0.04 0.30 0.11 0.29 0.08 0.22 0.04LUNGS 1.24 0.16 1.04 0.12 0.87 0.04 0.72 0.06 0.57 0.07HEART 0.78 0.11 0.65 0.05 0.55 0.06 0.45 0.05 0.34 0.02BLOOD 2.10 0.10 1.83 0.17 1.37 0.14 1.17 0.11 0.86 0.05TAIL 0.85 0.30 2.43 3.83 0.63 0.10 0.84 0.14 0.84 0.07BRAIN 0.18 0.04 0.14 0.03 0.09 0.02 0.08 0.01 0.05 0.01THYROID 1.11 0.22 1.14 0.36 0.92 0.10 0.70 0.24 0.63 0.04EYES 0.20 0.03 0.23 0.05 0.23 0.01 0.17 0.03 0.18 0.05PANCREAS 0.47 0.07 0.41 0.09 0.38 0.07 0.42 0.13 0.21 0.01THYMUS 0.38 0.03 0.34 0.03 0.32 0.06 0.25 0.04 0.20 0.06__________________________________________________________________________ .sup.a Australian Albino Wistar Rats, % Injected dose per gram (X .+-. SD). Average of 5 animals per time point. TABLE 4__________________________________________________________________________BIODISTRIBUTION OF .sup.99m TC COMPLEX OF NO. 4.sup.aMEAN SD MEAN SD MEAN SD MEAN SD MEAN SD30 SEC 1 MIN 3 MIN 5 MIN 10 MIN__________________________________________________________________________LIVER 6.83 0.51 8.03 0.95 7.89 0.57 7.39 1.20 4.90 1.06SPLEEN 0.93 0.12 0.80 0.05 0.42 0.05 0.32 0.05 0.27 0.05KIDNEYS 5.57 0.88 4.86 0.78 4.54 0.68 3.89 0.26 2.33 0.50MUSCLE 0.36 0.05 0.29 0.04 0.22 0.05 0.18 0.05 0.13 0.04SKIN 0.53 0.09 0.51 0.07 0.46 0.06 0.37 0.05 0.36 0.04BONE 0.61 0.10 0.54 0.05 0.32 0.06 0.26 0.05 0.19 0.04LUNGS 2.37 0.46 1.67 0.13 1.00 0.21 0.66 0.13 0.53 0.09HEART 1.78 0.32 1.26 0.07 0.65 0.07 0.48 0.09 0.36 0.10BLOOD 5.84 1.01 3.96 0.32 1.71 0.83 1.33 0.33 0.99 0.22URINE -- -- -- -- -- -- -- -- 0.08 0.05STOMACH 1.04 0.41 0.95 0.30 3.09 1.27 2.71 4.08 3.80 6.73TOT GIT 0.61 0.08 0.74 0.26 1.59 0.63 2.62 0.70 5.16 1.86TAIL 1.03 1.02 0.53 0.23 0.39 0.07 1.23 2.00 0.32 0.10BRAIN 0.43 0.04 0.26 0.05 0.12 0.01 0.08 0.02 0.06 0.06THYROID 3.49 0.83 2.28 0.25 1.11 0.63 0.89 0.24 0.79 0.18EYES 0.22 0.03 0.19 0.05 0.15 0.04 0.13 0.08 0.16 0.04PANCREAS 0.90 0.13 0.69 0.20 0.55 0.34 0.32 0.07 0.39 0.22THYMUS 0.66 0.08 0.49 0.08 0.31 0.02 0.22 0.03 0.19 0.63__________________________________________________________________________ .sup.a Australian Albino Wistar Rats, % Injected dose per gram (X .+-. SD). Average of 5 animals per time point. TABLE 5__________________________________________________________________________BIODISTRIBUTION OF .sup.99m TC COMPLEX OF NO. 5.sup.aMEAN SD MEAN SD MEAN SD MEAN SD MEAN SD30 SEC 1 MIN 3 MIN 5 MIN 10 MIN__________________________________________________________________________LIVER 1.11 0.16 0.97 0.16 0.83 0.13 0.79 0.07 0.69 0.07SPLEEN 0.83 0.12 0.98 0.10 0.83 0.08 0.83 0.05 0.72 0.03KIDNEYS 4.35 0.55 5.70 0.45 4.70 0.10 6.04 1.92 3.61 0.36MUSCLE 0.25 0.03 0.30 0.01 0.27 0.04 0.25 0.01 0.23 0.04SKIN 0.31 0.03 0.38 0.06 0.43 0.04 0.41 0.09 0.46 0.04BONE 0.37 0.02 0.45 0.02 0.42 0.03 0.37 0.08 0.36 0.03LUNGS 1.66 0.19 1.69 0.20 1.31 0.09 1.25 0.15 1.17 0.10HEART 1.81 0.22 1.62 0.18 1.24 0.09 1.15 0.11 1.06 0.17BLOOD 4.92 0.67 4.22 0.34 3.26 0.29 3.13 0.17 2.59 0.19TAIL 0.22 0.11 0.14 0.01 0.17 0.12 0.15 0.03 0.19 0.05BRAIN 0.20 0.02 0.22 0.04 0.16 0.03 0.17 0.02 0.14 0.03THYROID 1.85 0.31 2.43 0.67 1.59 0.46 1.73 0.45 1.71 0.29EYES 0.17 0.01 0.23 0.03 0.19 0.03 0.18 0.02 0.18 0.02PANCREAS 0.80 0.10 0.73 0.09 0.55 0.07 0.56 0.06 0.43 0.06THYMUS 0.63 0.12 0.56 0.08 0.45 0.03 0.54 0.13 0.39 0.05STOMACH 0.47 0.10 0.42 0.12 0.36 0.11 0.37 0.12 0.24 0.06TOT GIT 0.35 0.04 0.34 0.04 0.31 0.04 0.29 0.06 0.27 0.02__________________________________________________________________________ .sup.a Australian Albino Wistar Rats, % Injected dose per gram (X .+-. SD). Average of 5 animals per time point. TABLE 6______________________________________COMPOUND NO. 6.sup.a MEAN SD MEAN SD 3 MIN 5 MIN______________________________________LIVER 6.52 0.62 9.39 1.04SPLEEN 2.24 0.42 1.78 0.15KIDNEYS 24.05 3.93 18.35 0.89MUSCLE 1.81 0.33 1.26 0.08SKIN 3.96 0.41 3.47 0.39BONE 1.91 0.02 1.39 0.11LUNGS 6.38 1.04 4.66 0.39HEART 3.84 0.62 2.80 0.38BLOOD 10.65 0.87 7.83 0.54URINE 1.30 0.88 0.95 0.43STOMACH 1.78 0.38 6.85 6.36TOT GIT 1.03 0.05 2.12 0.38TAIL 8.94 3.42 6.18 2.50BRAIN 0.74 0.21 0.35 0.11THYROID 5.76 1.26 3.73 0.74PANCREAS 1.86 0.18 1.41 0.22THYMUS 3.67 1.03 2.50 0.41______________________________________ .sup.a Balb/c mice, % Injected dose per gram (X .+-. SD). Average of 5 animals per time point. TABLE 7__________________________________________________________________________Biodistribution of Copper-67 complex of (5)Biodistribution of Ligand (5)-67CuTimes 1', 3', 5', 10', 30'.Animals 25 Balb/c mice.Results: % Injected dose.% InjDose 1 min S.D. 3 min S.D. 5 min S.D. 10 min S.D. 30 min S.D.__________________________________________________________________________Liver 5.71 2.27 8.48 0.89 9.04 1.94 11.79 0.80 17.10 1.02Spleen 0.18 0.11 0.19 0.03 0.16 0.04 0.16 0.02 0.19 0.03Kidney 8.35 3.82 5.65 0.81 4.99 0.99 4.48 0.42 3.38 0.35Muscle 13.69 9.74 18.00 3.97 11.95 2.94 8.38 1.81 7.45 4.44Skin 9.61 3.87 19.21 1.31 16.23 3.68 16.46 2.03 14.14 3.19Bone 5.46 1.97 3.35 0.50 2.74 0.57 3.2 1 0.80 2.05 0.16Lung 0.85 0.38 1.20 0.27 1.10 0.21 1.20 0.22 1.29 0.41Heart 0.27 0.09 0.28 0.05 0.19 0.03 0.18 0.03 0.20 0.04Blood 5.86 3.44 4.76 2.07 4.05 0.63 2.33 0.49 2.12 0.26Urine 0.35 0.47 1.98 1.33 3.72 3.97 4.71 2.87 1.33 1.13Stomach 0.52 0.22 0.93 0.15 0.87 0.26 0.80 0.41 1.22 0.26Tail 8.27 9.95 5.19 4.72 5.36 2.39 3.45 2.23 1.51 0.47Pancreas 0.20 0.13 0.11 0.03 0.12 0.04 0.34 0.22 0.34 0.05Brain 0.23 0.12 0.22 0.02 0.23 0.09 0.19 0.15 0.54 0.88S. Int. 0.50 0.23 1.10 0.23 1.08 0.24 0.90 0.48 0.71 0.32L. Int. 0.35 0.14 0.75 0.26 0.54 0.15 0.79 0.25 0.31 0.15__________________________________________________________________________ TABLE 8__________________________________________________________________________Biodistribution of .sup.99m Tc complex of compound 1.MEAN S.D. MEAN S.D. MEAN S.D. MEAN S.D. MEAN S.D.ORGAN 1 min 3 min 5 min 10 min 30 min__________________________________________________________________________Liver 12.07 0.43 12.83 0.86 11.82 0.77 5.96 3.53 3.09 1.1Spleen 0.17 0.01 0.1 0.01 0.07 0.02 0.06 0.01 0.02 0.01Kidney 11.78 2.27 11.23 1.4 9.17 2.29 5.58 1.43 3.19 0.28Muscle 23.48 4.91 13.18 4.19 9.81 2.24 16.44 13.81 4.98 3.91Skin 19.66 3.67 19.28 1.67 19.68 3.48 15.68 2.13 7.31 5.21Bone 5.34 0.42 3.1 0.7 2.88 0.4 5.06 3.58 1.12 0.41Lungs 1.54 0.48 0.98 0.31 0.77 0.13 0.48 0.13 0.2 0.06Heart 0.57 0.1 0.23 0.05 0.21 0.03 0.12 0.02 0.05 0Blood 27.09 3.45 12.02 1.57 9.77 0.73 5.34 0.74 1.82 0.3Urine 0 0 5.62 1.01 4.51 5.04 11.64 5.79 8.84 5.77Bladder 0.12 0.02 0.63 0.42 0.68 0.21 1.96 1.71 1.23 2.01Stomach 0.5 0.09 0.98 0.21 1.02 0.26 1.16 0.69 0.49 0.31GIT 3.82 0.88 6.06 0.59 4.77 2.1 12.35 1.63 20 0.9Tail 2.42 1.4 1.58 0.68 3.76 2.12 2.07 0.69 1.53 1.1__________________________________________________________________________ Balb/c mice, % Injected dose (X .+-. SD), Average of 5 animals per time point. TABLE 9__________________________________________________________________________Biodistribution of .sup.99m Tc complex of compound 2.MEAN S.D. MEAN S.D. MEAN S.D. MEAN S.D. MEAN S.D.ORGAN 1 min 3 min 5 min 10 min 30 min__________________________________________________________________________Liver 14.68 0.57 14.52 2.24 7.75 4.13 7.95 1.15 5.71 1.53Spleen 0.19 0.02 0.16 0.03 0.07 0.04 0.06 0.01 0.05 0.01Kidney 8.04 1.53 10.44 1.83 5.45 3.53 5.14 0.73 4 0.5Muscle 19.67 5.83 14.89 4.78 6.73 3.57 8.73 3.93 5.48 2.78Skin 13.37 0.85 13.1 0.89 7.96 4.6 11.02 3.76 5.58 1.27Bone 7.88 1.45 6.29 0.87 2.99 1.42 2.76 0.4 2.27 0.53Lungs 1.36 0.28 1.19 0.27 0.72 0.3 0.53 0.07 0.43 0.15Heart 0.49 0.07 0.34 0.08 0.22 0.09 0.15 0.01 0.09 0.01Blood 23.14 3.9 14.96 2.56 7.4 3.51 7.15 0.55 3.58 0.24Urine 0.09 0.03 2.78 3.55 1.06 0.95 2.67 2.31 3.84 1.08BladderStomach 0.67 0.09 0.998 0.07 0.6 0.34 1.3 0.26 2.06 0.56GIT 3.74 0.52 4.86 0.8 3.38 2.46 8.11 4.14 16.24 5.2Tail 4.05 3.97 6.89 5.52 30.12 14.89 18.31 8.6 7.67 4.69__________________________________________________________________________ Balb/c mice, % Injected dose (X .+-. SD), Average of 5 animals per time point. TABLE 10__________________________________________________________________________Biodistribution of .sup.99m Tc complex of compound 4.MEAN S.D. MEAN S.D. MEAN S.D. MEAN S.D. MEAN S.D.ORGAN 1 min 3 min 5 min 10 min 30 min__________________________________________________________________________Liver 31.82 3.89 40.23 4.84 30.24 3.75 21.19 3.76 12.48 2.01Spleen 0.21 0.03 0.12 0.03 0.1 0.02 0.07 0.03 0.06 0.02Kidney 7.83 1.04 6.27 0.57 4.18 0.73 3.57 1.09 3.67 0.36Muscle 11.85 5.14 9.92 0.68 7.77 2.09 4.95 1.66 3.35 1.39Skin 8.94 1.18 9.02 0.74 8.67 0.53 7.3 0.55 4 0.33Bone 6.4 1.53 2.5 0.5 2.84 1.71 2.08 0.74 1.67 0.98Lungs 1.29 0.85 0.56 0.16 0.68 0.25 0.52 0.16 0.26 0.11Heart 0.6 0.13 0.22 0.02 0.16 0.04 0.13 0.03 0.05 0.01Blood 34.01 4.04 12.06 1.89 7.87 0.38 5.04 0.26 3.88 1.01Urine 0 0 0.84 0.47 1.55 0.73 4.27 2.46 3.21 3.26Bladder 0.06 0.04 0.09 0.07 0.04 0.03 0.17 0.09 0.08 0.1Stomach 0.52 0.07 0.61 0.25 3.96 2.59 0.7 0.52 4.84 4.82GIT 3.96 1.76 8.34 2.99 13.7 5.33 33.63 3.55 39.97 10.4Tail 4.62 1.56 2.02 0.74 4.63 4.59 3.58 3.36 1.91 0.74__________________________________________________________________________ Balb/c mice, % Injected dose (X .+-. SD), Average of 5 animals per time point. TABLE 11__________________________________________________________________________Biodistribution of .sup.99m Tc complex of compound 5.MEAN S.D. MEAN S.D. MEAN S.D. MEAN S.D. MEAN S.D.ORGAN 1 min 3 min 5 min 10 min 30 min__________________________________________________________________________Liver 5.72 0.55 5.04 0.7 4.34 0.84 5.34 0.82 2.86 0.37Spleen 0.14 0.04 0.16 0.02 0.12 0.02 0.11 0.01 0.06 0.01Kidney 9.08 2.1 8.71 1.52 6.71 1.87 5.97 0.71 3.75 0.66Muscle 23.78 3.73 15.39 1.79 11.74 2.25 9.67 1.93 23.84 33.4Skin 18.09 4.65 19.66 4.38 19.83 6.59 23.93 4.64 17.37 6.21Bone 6.02 1.8 4 0.29 2.79 0.32 2.98 0.54 1.86 0.82Lungs 1.64 0.29 1.65 0.7 1.28 0.32 1.1 0.36 0.46 0.1Heart 0.61 0.1 0.43 0.03 0.26 0.06 0.23 0.05 0.12 0.05Blood 26.59 1.56 14.84 1.03 10.06 1.33 12.87 8.12 3.98 0.56Urine 0 0 2.97 1.49 6.52 4.77 10.66 8.48 5.91 3.45Bladder 0.09 0.05 1.16 0.43 2.15 1.62 1.04 0.97 0.83 0.99Stomach 0.65 0.11 0.56 0.09 0.4 0.08 0.63 0.08 0.49 0.35GIT 3.89 0.86 3.22 0.08 2.42 0.53 3.94 0.75 4.61 0.75Tail 2.65 0.84 4.28 4.1 11.55 12.68 6.98 6.75 2.01 0.6__________________________________________________________________________ Balb/c mice, % Injected dose (X .+-. SD), Average of 5 animals per time point. TABLE 12__________________________________________________________________________Biodistribution of .sup.99m Tc complex of compound 6.MEAN S.D. MEAN S.D. MEAN S.D. MEAN S.D. MEAN S.D.ORGAN 1 min 3 min 5 min 10 min 30 min__________________________________________________________________________Liver 22.44 3.45 35.66 2.09 30.85 6 16.04 1.79 14.05 2.58Spleen 0.2 0.04 0.09 0.03 0.14 0.05 0.07 0.02 0.06 0.03Kidney 7.57 1.71 5.68 0.79 4.53 0.57 2.44 0.34 1.9 0.19Muscle 13.95 2.12 7.87 2.49 7.24 1.79 5.27 2.52 6.92 8.06Skin 8.13 1.14 8.31 1.33 9.28 3.33 5.17 1.92 4.23 2.25Bone 7.37 0.86 3.5 1.17 3.86 0.52 2.76 1.63 2.54 1.37Lungs 1.71 0.72 0.65 0.33 0.69 0.19 0.45 0.13 0.39 0.09Heart 0.46 0.04 0.2 0.04 0.16 0.02 0.08 0.04 0.11 0.09Blood 41.63 3.94 15.59 1.22 10.31 0.77 6.34 1.8 9.66 14.1Urine 0.07 0.15 0.81 0.52 0.59 0.49 1.231 0.59 1.64 0.74Bladder 0.06 0.01 0.08 0.05 0.07 0.01 0.23 0.19 0.2 0.14Stomach 0.4 0.06 0.43 0.21 5.42 5.84 3.42 4.01 1.73 1.39GIT 3.58 1.37 11.98 7.7 27.85 7.9 45.71 6.16 51.18 3.62Tail 8.46 7.08 7.37 4.71 7.64 5.59 4.77 2.15 2.41 3.11__________________________________________________________________________ Balb/c mice, % Injected dose (X .+-. SD), Average of 5 animals per time point. TABLE 13__________________________________________________________________________BIODISTRIBUTION OF GA-67 (1) 1 MIN 3 MIN 5 MIN 10 MIN 30 MIN 120 MINORGAN MEAN S.D. MEAN S.D. MEAN S.D. MEAN S.D. MEAN S.D. MEAN S.D.__________________________________________________________________________LIVER 15.45 1.65 24.21 1.93 25.79 1.7 22.77 1.74 10.11 1.15 0.88 0.56SPLEEN 0.29 0.05 0.14 0.02 0.1 0.01 0.06 0.02 0.01 0.01 0.01 0KIDNEY 9.28 1.50 5.26 2.06 4.37 1.97 1.91 0.33 0.57 0.12 0.05 0.01MUSCLE 22.75 2.64 12.70 1.58 12.14 6.29 26.13 39.89 1.80 1.44 0.48 0.26SKIN 11.72 0.62 13.76 1.63 13.02 1.63 8.47 0.99 1.56 0.52 0.24 0.09BONE 5.07 0.42 3.30 0.57 2.83 0.71 2.20 1.04 0.50 0.22 0.42 0.22LUNGS 1.28 0.42 0.83 0.17 1.11 0.49 0.46 0.12 0.12 0.06 0.04 0.02HEART 0.41 0.07 0.19 0.02 0.15 0.03 0.08 0.01 0.01 0.00 0.01 0.00BLOOD 16.86 1.74 9.81 0.34 7.82 1.59 3.52 0.55 0.78 0.41 0.35 0.21BLADDER 0.08 0.02 0.11 0.03 0.09 0.06 0.09 0.07 0.03 0.03 0.01 0.01STOMACH SQ 0.08 0.03 0.08 0.02 0.11 0.05 0.10 0.08 0.02 0.01 0.02 0.02STOMACH GL 0.36 0.05 0.19 0.10 0.21 0.08 0.18 0.12 0.04 0.04 0.03 0.03DUODENUM 0.12 0.05 0.09 0.04 0.17 0.13 0.24 0.13 0.06 0.05 0.02 0.01JEJUNUM 1.27 0.22 0.69 0.16 0.73 0.28 1.73 0.50 2.99 1.09 0.85 0.47ILEUM 0.25 0.12 0.13 0.06 0.11 0.07 0.18 0.18 0.11 0.13 0.66 0.65CAECUM 0.45 0.18 0.26 0.06 0.22 0.06 0.17 0.13 0.06 0.04 0.88 0.53COLON 0.53 0.13 0.43 0.13 0.37 0.10 0.21 0.10 0.08 0.04 0.24 0.14TAIL 1.13 0.31 1.50 0.32 1.95 0.76 1.48 0.28 1.06 0.46 0.10 0.08PANCREAS 0.35 0.11 0.16 0.05 0.11 0.01 0.07 0.03 0.02 0.01 0.01 0.00SALIVARY GL 0.16 0.04 0.11 0.04 0.10 0.05 0.07 0.03 0.01 0.01 0.01 0.01OVARIES 0.07 0.03 0.06 0.02 0.07 0.02 0.03 0.01 0.01 0.00 0.00 0.00__________________________________________________________________________ .sup.a Balb/c mice, % ID (X +/- S.D.). Average of 5 animals per timepoint TABLE 14__________________________________________________________________________BIODISTRIBUTION OF GA-67 (2) 1 MIN 3 MIN 5 MIN 10 MIN 30 MIN 120 MINORGAN MEAN S.D. MEAN S.D. MEAN S.D. MEAN S.D. MEAN S.D. MEAN S.D.__________________________________________________________________________LIVER 24.9 5.47 35.52 3.34 33.09 3.19 23.34 4.55 3.7 1.22 0.92 0.86SPLEEN 0.29 0.1 0.1 0.063 0.07 0.01 0.02 0.01 0.02 0.02 0 0KIDNEY 9.54 0.93 4.62 0.93 3.52 0.81 1.00 0.17 0.21 0.05 0.01 0.01MUSCLE 20.55 5.85 8.63 1.46 5.50 1.60 2.00 0.68 2.00 3.54 0.31 0.38SKIN 11.55 1.74 9.27 1.08 8.04 1.05 3.80 1.07 0.88 0.64 0.17 0.17BONE 6.16 1.15 2.82 0.42 2.13 0.36 0.74 0.12 0.50 0.54 0.38 0.61LUNGS 1.79 0.69 0.56 0.12 0.41 0.11 0.12 0.06 0.06 0.06 0.01 0.01HEART 0.42 0.09 0.14 0.03 0.09 0.03 0.03 0.00 0.01 0.01 0.00 0.00BLOOD 17.45 4.46 6.02 0.86 3.31 0.70 2.16 2.31 3.20 4.40 0.07 0.13BLADDER 0.08 0.02 0.06 0.01 0.09 0.03 0.03 0.01 0.01 0.01 0.00 0.00STOMACH SQ. 0.07 0.03 0.08 0.03 0.18 0.28 0.25 0.40 0.03 0.05 0.03 0.03STOMACH GL. 0.26 0.12 0.20 0.08 0.53 0.72 0.28 0.28 0.11 0.21 0.03 0.04DUODENUM 0.13 0.04 0.34 0.31 0.67 0.26 1.22 1.24 0.31 0.15 0.04 0.06JEJUNEM 1.42 0.42 1.67 0.82 3.68 1.80 5.99 5.13 6.64 4.73 0.99 1.06ILEUM 0.15 0.07 0.11 0.09 0.21 0.20 0.03 0.01 0.78 1.49 0.28 0.26CAECUM 0.42 0.14 0.23 0.10 0.15 0.06 0.04 0.01 0.20 0.24 7.06 7.34COLON 0.48 0.17 0.28 0.07 0.25 0.11 0.10 0.02 0.16 0.08 2.03 2.01PANCREAS 0.34 0.14 0.11 0.03 0.10 0.06 0.03 0.01 0.08 0.14 0.00 0.00SALIVARY GL. 0.11 0.06 0.05 0.01 0.02 0.01 0.01 0.00 0.00 0.00 0.00 0.00OVARIES 0.06 0.01 0.05 0.01 0.04 0.01 0.01 0.00 0.01 0.02 0.00 0.00TAIL 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0.56 0.13 0.12 0.06__________________________________________________________________________ .sup.a Balb/c mice, % ID (X +/- S.D.). Average of 5 animals per timepoint TABLE 15__________________________________________________________________________BIODISTRIBUTION OF GA-67 (4) 1 MIN 3 MIN 5 MIN 10 MIN 30 MIN 120 MINORGAN MEAN S.D. MEAN S.D. MEAN S.D. MEAN S.D. MEAN S.D. MEAN S.D.__________________________________________________________________________LIVER 39.06 1.27 43.23 1.17 39.61 2.10 8.29 2.00 1.06 0.96 0.29 0.18SPLEEN 0.15 0.03 0.04 0.02 0.03 0.00 0.00 0.00 0.00 0.00 0.01 0.00KIDNEY 6.91 0.97 2.12 0.26 1.59 0.15 0.43 0.05 0.17 0.09 0.07 0.01MUSCLE 11.34 3.66 5.08 1.98 4.29 0.65 0.65 0.38 0.32 0.28 0.21 0.09SKIN 11.94 0.78 8.12 0.66 8.22 1.18 2.92 3.85 0.18 0.12 0.15 0.02BONE 3.20 0.57 1.31 0.09 0.98 0.20 0.20 0.12 0.13 0.04 0.15 0.05LUNGS 0.64 0.21 0.26 0.05 0.18 0.07 0.03 0.00 0.01 0.00 0.01 0.00HEART 0.27 0.06 0.08 0.01 0.05 0.02 0.01 0.00 0.00 0.00 0.00 0.00BLOOD 9.95 2.46 3.06 0.41 2.28 0.50 0.27 0.04 0.13 0.03 0.10 0.02BLADDER 0.14 0.13 0.28 0.24 0.14 0.09 0.41 0.57 0.00 0.01 0.00 0.00STOMACH SQ. 0.06 0.03 0.03 0.01 0.07 0.07 0.04 0.08 0.01 0.02 0.21 0.42STOMACH GL. 0.14 0.03 0.06 0.02 0.47 0.48 0.19 0.35 0.02 0.01 0.02 0.02DUODENUM 0.09 0.03 0.25 0.27 0.44 0.32 0.36 0.37 0.53 1.16 0.04 0.05JEJUNEM 1.16 0.28 1.17 0.32 1.19 0.59 5.43 1.56 0.61 0.38 0.20 0.18ILEUM 0.20 0.07 0.080 0.03 0.05 0.02 0.06 0.04 01.52 1.97 0.12 0.09CAECUM 0.19 0.10 0.15 0.05 0.06 0.01 0.09 0.03 0.49 0.12 1.19 0.33COLON 0.32 0.09 0.11 0.03 0.14 0.07 0.06 0.04 0.28 0.25 0.56 0.34TAIL 0.00 0.00 0.88 0.17 0.73 0.25 0.57 0.11 0.14 0.04 0.11 0.06PANCREAS 0.17 0.05 0.02 0.10 0.14 0.03 0.04 0.00 0.00 0.01 0.01SALIVARY GL. 0.12 0.08 0.06 0.03 0.03 0.02 0.01 0.02 0.00 0.00 0.00 0.01OVARIES 0.13 0.03 0.12 0.06 0.10 0.03 0.02 0.01 0.00 0.00 0.00 0.00__________________________________________________________________________ .sup.a Balb/c mice, % ID (X +/- S.D.). Average of 5 animals per timepoint TABLE 16__________________________________________________________________________BIODISTRIBUTION OF GA-67 (5) 1 MIN 3 MIN 5 MIN 10 MIN 30 MIN 120 MINORGAN MEAN S.D. MEAN S.D. MEAN S.D. MEAN S.D. MEAN S.D. MEAN S.D.__________________________________________________________________________LIVER 3.08 0.29 3.02 0.41 2.04 0.33 1.22 0.15 1.33 0.32 0.86 0.12SPLEEN 0.18 0.03 0.15 0.01 0.10 0.02 0.05 0.02 0.03 0.01 0.02 0.00KIDNEY 8.39 2.08 5.78 0.54 3.17 0.17 2.39 0.31 1.72 0.26 1.60 0.16MUSCLE 23.50 9.80 19.47 7.65 20.11 19.15 4.58 2.05 2.65 1.01 1.44 0.77SKIN 20.79 1.42 21.98 1.64 18.72 1.68 10.65 1.90 5.12 0.75 2.82 0.28BONE 4.85 0.31 5.90 2.14 4.51 1.59 21.3 0.65 1.69 1.72 0.80 0.45LUNGS 1.15 0.22 0.89 0.10 0.76 0.22 0.22 0.03 0.16 0.08 0.10 0.03HEART 0.35 0.03 0.27 0.03 0.19 0.02 0.07 0.01 0.04 0.01 0.03 0.01BLOOD 17.48 0.91 12.82 0.86 8.06 0.54 2.86 0.19 1.16 0.19 0.65 0.07URINE 6.30 3.49 8.94 5.16 15.34 3.12 14.23 4.14 7.76 2.85 3.55 2.07BLADDER 0.48 0.54 0.62 1.01 0.47 0.43 0.10 0.04 1.04 1.90 0.32 0.57STOMACH SQ. 0.08 0.02 0.10 0.01 0.10 0.07 0.03 0.01 0.02 0.01 0.01 0.00STOMACH GL. 0.27 0.05 0.17 0.04 0.11 0.03 0.04 0.01 0.03 0.01 0.02 0.01DUODENUM 0.12 0.02 0.10 0.01 0.05 0.02 0.03 0.02 0.02 0.01 0.01 0.00JEJUNUM 0.81 0.35 0.61 0.17 0.46 0.11 0.29 0.06 0.18 0.03 0.09 0.02ILEUM 0.32 0.12 0.20 0.10 0.18 0.07 0.08 0.07 0.07 0.02 0.06 0.03CAECUM 0.36 0.08 0.26 0.06 0.31 0.35 0.09 0.06 0.04 0.01 0.03 0.02COLON 0.59 0.15 0.53 0.09 0.37 0.12 0.11 0.04 0.05 0.01 0.05 0.01TAIL 0.86 0.19 0.64 0.17PANCREAS 0.36 0.20 0.19 0.04 0.16 0.03 0.05 0.01 0.05 0.04 0.01 0.00SALIV 0.29 0.06 0.28 0.04 0.16 0.05 0.06 0.01 0.04 0.01 0.03 0.02TESTIES 0.25 0.05 0.25 0.07 0.32 0.20 0.08 0.01 0.10 0.05 0.05 0.03__________________________________________________________________________ .sup.a Balb/c mice, % ID (X +/- S.D.). Average of 5 animals per timepoint TABLE 17__________________________________________________________________________BIODISTRIBUTION OF GA-67 (6) 1 MIN 3 MIN 5 MIN 10 MIN 30 MIN 120 MINORGAN MEAN S.D. MEAN S.D. MEAN S.D. MEAN S.D. MEAN S.D. MEAN S.D.__________________________________________________________________________LIVER 26.45 1.01 34.06 1.45 30.21 2.02 18.58 2.52 3.15 1.92 1.33 1.88SPLEEN 0.22 0.02 0.08 0.03 0.04 0.02 0.03 0.01 0.01 0.00 0.00 0.00KIDNEY 9.95 0.52 5.24 1.09 3.36 0.50 1.98 0.16 0.79 0.09 0.06 0.01MUSCLE 17.93 3.63 10.67 1.25 6.35 1.58 4.37 1.32 7.73 11.64 1.85 3.85SKIN 13.91 1.44 12.80 1.65 10.62 1.41 5.76 0.93 0.86 0.24 0.10 0.05BONE 4.33 1.43 2.41 0.34 1.67 0.31 0.96 0.62 0.34 0.14 0.06 0.04LUNGS 0.78 0.17 0.64 0.37 0.25 0.06 0.17 0.08 0.19 0.36 0.01 0.01HEART 0.29 0.02 0.13 0.02 0.09 0.03 0.04 0.01 0.01 0.00 0.00 0.00BLOOD 13.61 2.51 6.08 0.94 3.02 0.60 1.44 0.14 0.33 0.25 1.79 3.93BLADDER 0.33 0.31 0.62 0.45 0.94 1.13 0.37 0.59 0.27 0.45 0.62 1.39STOMACH SQ. 0.08 0.06 0.07 0.03 0.04 0.02 0.32 0.53 0.03 0.02 0.01 0.00STOMACH GL. 0.20 0.12 0.22 0.11 0.20 0.21 1.61 1.28 0.35 0.43 0.01 0.01DUODENUM 0.08 0.02 0.43 0.64 0.41 0.22 1.75 0.63 0.27 0.26 0.02 0.02JEJUNEM 1.19 0.26 1.25 0.72 2.25 1.48 5.34 3.57 3.41 1.70 0.12 0.07ILEUM 0.21 0.07 0.15 0.16 0.08 0.02 0.13 0.17 0.06 0.06 0.10 0.06CAECUM 0.27 0.09 0.20 0.06 0.13 0.02 0.17 0.07 0.15 0.20 1.70 0.74COLON 0.49 0.11 0.28 0.09 0.18 0.05 0.27 0.31 0.27 0.28 0.72 0.35TAIL 0.73 0.00 0.84 0.10 0.55 0.31 0.10 0.09PANCREAS 0.25 0.11 0.11 0.02 0.06 0.03 0.12 0.17 0.08 0.14 0.00 0.00SALIVARY GL 0.18 0.16 0.12 0.05 0.06 0.04 0.02 0.01 0.01 0.01 0.00 0.00TESTES 0.17 0.03 0.17 0.04 0.12 0.03 0.07 0.02 0.07 0.05 0.02 0.04__________________________________________________________________________ .sup.a Balb/c mice, % ID (X +/- S.D.). Average of 5 animals per timepoint

Claims

1. A physiologically acceptable, radiolabelled complex consisting of a complex of a radionuclide and a compound of the formula ##STR11## where k is an integer from 2 to 5

l is an integer from 1 to 5

X and Y are independently selected from phenyl, naphthyl, pyridine or quinoline radical, each having at position 2 one of --OH, --SH, --NH.sub.2 or --COOH and each of X and Y can be optionally substituted by one or more amino, halogen, hydroxy, mercapto, nitro, cyano, thiocyano, alkyl, alkoxy, halogenoalkyl, acyl, acylamino, acyloxy, carboxyl, alkoxycarbonyl, carbamoyl, pyridoylamino, N-carboxyalkyl-carbamoyl, sulpho, sulphamoyl, mono- or dialkylated or phenylated sulphamoyl which can also carry one or more substituents R', alkylsulphonyl, alkoxysulphonyl, or by an optionally hydroxy-containing phenylsulphonyl or phenoxysulphonyl; where R' is as defined for X and Y; or pharmaceutically acceptable salts thereof.

2. The complex according to claim 1 where k is 2 and l is the integer 1.

3. A diagnostic formulation suitable for labelling iwith .sup.99m Tc, .sup.188 Re or .sup.186 Re comprising a compound of formula (I) as defined in claim 1 or a pharmaceutically acceptable salt thereof and a reducing agent in a pharmaceutically acceptable carrier.

4. A formulation comprising a radiolabelled complex or formula (I) as defined in claim 1 or a pharmaceutically acceptable salt thereof in a pharmaceutically acceptable carrier.

5. A method of diagnosis or therapy in a subject comprising administering to the subject a radiolabelled complex of formula (I) as defined in claim 1 or a pharmaceutically acceptable salt thereof.

6. The method according to claim 5 where in the complex of formula (I), k is 2 and l is the integer 1.