CA2062454A1 - Amide-chelates, their metal complexes as well as their use in diagnosis and treatment - Google Patents

Amide-chelates, their metal complexes as well as their use in diagnosis and treatment

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CA2062454A1
CA2062454A1 CA002062454A CA2062454A CA2062454A1 CA 2062454 A1 CA2062454 A1 CA 2062454A1 CA 002062454 A CA002062454 A CA 002062454A CA 2062454 A CA2062454 A CA 2062454A CA 2062454 A1 CA2062454 A1 CA 2062454A1
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Reinhard Neumeier
Gerhard Rohlfs
Wolfgang Kramp
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Bayer Pharma AG
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    • C07D213/00—Heterocyclic compounds containing six-membered rings, not condensed with other rings, with one nitrogen atom as the only ring hetero atom and three or more double bonds between ring members or between ring members and non-ring members
    • C07D213/02—Heterocyclic compounds containing six-membered rings, not condensed with other rings, with one nitrogen atom as the only ring hetero atom and three or more double bonds between ring members or between ring members and non-ring members having three double bonds between ring members or between ring members and non-ring members
    • C07D213/04—Heterocyclic compounds containing six-membered rings, not condensed with other rings, with one nitrogen atom as the only ring hetero atom and three or more double bonds between ring members or between ring members and non-ring members having three double bonds between ring members or between ring members and non-ring members having no bond between the ring nitrogen atom and a non-ring member or having only hydrogen or carbon atoms directly attached to the ring nitrogen atom
    • C07D213/60—Heterocyclic compounds containing six-membered rings, not condensed with other rings, with one nitrogen atom as the only ring hetero atom and three or more double bonds between ring members or between ring members and non-ring members having three double bonds between ring members or between ring members and non-ring members having no bond between the ring nitrogen atom and a non-ring member or having only hydrogen or carbon atoms directly attached to the ring nitrogen atom with hetero atoms or with carbon atoms having three bonds to hetero atoms with at the most one bond to halogen, e.g. ester or nitrile radicals, directly attached to ring carbon atoms
    • C07D213/62—Oxygen or sulfur atoms
    • C07D213/70—Sulfur atoms
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K47/00—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient
    • A61K47/50—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates
    • A61K47/51—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates the non-active ingredient being a modifying agent
    • A61K47/68—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates the non-active ingredient being a modifying agent the modifying agent being an antibody, an immunoglobulin or a fragment thereof, e.g. an Fc-fragment
    • A61K47/6835—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates the non-active ingredient being a modifying agent the modifying agent being an antibody, an immunoglobulin or a fragment thereof, e.g. an Fc-fragment the modifying agent being an antibody or an immunoglobulin bearing at least one antigen-binding site
    • A61K47/6887—Antibody-chelate conjugates using chelates for therapeutic purposes
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P35/00—Antineoplastic agents
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07C—ACYCLIC OR CARBOCYCLIC COMPOUNDS
    • C07C233/00—Carboxylic acid amides
    • C07C233/01—Carboxylic acid amides having carbon atoms of carboxamide groups bound to hydrogen atoms or to acyclic carbon atoms
    • C07C233/16—Carboxylic acid amides having carbon atoms of carboxamide groups bound to hydrogen atoms or to acyclic carbon atoms having the nitrogen atom of at least one of the carboxamide groups bound to a carbon atom of a hydrocarbon radical substituted by singly-bound oxygen atoms
    • C07C233/17—Carboxylic acid amides having carbon atoms of carboxamide groups bound to hydrogen atoms or to acyclic carbon atoms having the nitrogen atom of at least one of the carboxamide groups bound to a carbon atom of a hydrocarbon radical substituted by singly-bound oxygen atoms with the substituted hydrocarbon radical bound to the nitrogen atom of the carboxamide group by an acyclic carbon atom
    • C07C233/20—Carboxylic acid amides having carbon atoms of carboxamide groups bound to hydrogen atoms or to acyclic carbon atoms having the nitrogen atom of at least one of the carboxamide groups bound to a carbon atom of a hydrocarbon radical substituted by singly-bound oxygen atoms with the substituted hydrocarbon radical bound to the nitrogen atom of the carboxamide group by an acyclic carbon atom having the carbon atom of the carboxamide group bound to a carbon atom of an acyclic unsaturated carbon skeleton
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07C—ACYCLIC OR CARBOCYCLIC COMPOUNDS
    • C07C237/00—Carboxylic acid amides, the carbon skeleton of the acid part being further substituted by amino groups
    • C07C237/02—Carboxylic acid amides, the carbon skeleton of the acid part being further substituted by amino groups having the carbon atoms of the carboxamide groups bound to acyclic carbon atoms of the carbon skeleton
    • C07C237/20—Carboxylic acid amides, the carbon skeleton of the acid part being further substituted by amino groups having the carbon atoms of the carboxamide groups bound to acyclic carbon atoms of the carbon skeleton the carbon skeleton containing six-membered aromatic rings
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07C—ACYCLIC OR CARBOCYCLIC COMPOUNDS
    • C07C323/00—Thiols, sulfides, hydropolysulfides or polysulfides substituted by halogen, oxygen or nitrogen atoms, or by sulfur atoms not being part of thio groups
    • C07C323/23—Thiols, sulfides, hydropolysulfides or polysulfides substituted by halogen, oxygen or nitrogen atoms, or by sulfur atoms not being part of thio groups containing thio groups and nitrogen atoms, not being part of nitro or nitroso groups, bound to the same carbon skeleton
    • C07C323/39—Thiols, sulfides, hydropolysulfides or polysulfides substituted by halogen, oxygen or nitrogen atoms, or by sulfur atoms not being part of thio groups containing thio groups and nitrogen atoms, not being part of nitro or nitroso groups, bound to the same carbon skeleton at least one of the nitrogen atoms being part of any of the groups, X being a hetero atom, Y being any atom
    • C07C323/40—Y being a hydrogen or a carbon atom
    • C07C323/41—Y being a hydrogen or an acyclic carbon atom
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07C—ACYCLIC OR CARBOCYCLIC COMPOUNDS
    • C07C323/00—Thiols, sulfides, hydropolysulfides or polysulfides substituted by halogen, oxygen or nitrogen atoms, or by sulfur atoms not being part of thio groups
    • C07C323/50—Thiols, sulfides, hydropolysulfides or polysulfides substituted by halogen, oxygen or nitrogen atoms, or by sulfur atoms not being part of thio groups containing thio groups and carboxyl groups bound to the same carbon skeleton
    • C07C323/51—Thiols, sulfides, hydropolysulfides or polysulfides substituted by halogen, oxygen or nitrogen atoms, or by sulfur atoms not being part of thio groups containing thio groups and carboxyl groups bound to the same carbon skeleton having the sulfur atoms of the thio groups bound to acyclic carbon atoms of the carbon skeleton
    • C07C323/52—Thiols, sulfides, hydropolysulfides or polysulfides substituted by halogen, oxygen or nitrogen atoms, or by sulfur atoms not being part of thio groups containing thio groups and carboxyl groups bound to the same carbon skeleton having the sulfur atoms of the thio groups bound to acyclic carbon atoms of the carbon skeleton the carbon skeleton being acyclic and saturated
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07C—ACYCLIC OR CARBOCYCLIC COMPOUNDS
    • C07C331/00—Derivatives of thiocyanic acid or of isothiocyanic acid
    • C07C331/16—Isothiocyanates
    • C07C331/28—Isothiocyanates having isothiocyanate groups bound to carbon atoms of six-membered aromatic rings
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D495/00—Heterocyclic compounds containing in the condensed system at least one hetero ring having sulfur atoms as the only ring hetero atoms
    • C07D495/02—Heterocyclic compounds containing in the condensed system at least one hetero ring having sulfur atoms as the only ring hetero atoms in which the condensed system contains two hetero rings
    • C07D495/04—Ortho-condensed systems
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07J—STEROIDS
    • C07J41/00—Normal steroids containing one or more nitrogen atoms not belonging to a hetero ring
    • C07J41/0033—Normal steroids containing one or more nitrogen atoms not belonging to a hetero ring not covered by C07J41/0005
    • C07J41/0044—Normal steroids containing one or more nitrogen atoms not belonging to a hetero ring not covered by C07J41/0005 with an estrane or gonane skeleton, including 18-substituted derivatives and derivatives where position 17-beta is substituted by a carbon atom not directly bonded to another carbon atom and not being part of an amide group
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07J—STEROIDS
    • C07J41/00—Normal steroids containing one or more nitrogen atoms not belonging to a hetero ring
    • C07J41/0033—Normal steroids containing one or more nitrogen atoms not belonging to a hetero ring not covered by C07J41/0005
    • C07J41/0072—Normal steroids containing one or more nitrogen atoms not belonging to a hetero ring not covered by C07J41/0005 the A ring of the steroid being aromatic

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  • Biomedical Technology (AREA)
  • General Chemical & Material Sciences (AREA)
  • Bioinformatics & Cheminformatics (AREA)
  • Epidemiology (AREA)
  • Medicines Containing Antibodies Or Antigens For Use As Internal Diagnostic Agents (AREA)
  • Pyridine Compounds (AREA)
  • Pyrrole Compounds (AREA)
  • Heterocyclic Carbon Compounds Containing A Hetero Ring Having Oxygen Or Sulfur (AREA)
  • Medicines That Contain Protein Lipid Enzymes And Other Medicines (AREA)
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  • Organic Low-Molecular-Weight Compounds And Preparation Thereof (AREA)
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Abstract

Abstract The invention relates to compounds of general formula I

Description

20~2~
Amide-Chelates, their Metal CompleXes as well as their Use in Diagnosis and Treatment Description Radioactive metal ions, mostly bound to a complexing agent, have for some time been used for in vivo diagnosis. Among them, teehnetium-99m (Tc-99m), because of its almost ideal physieal properties for this purpose -- good absorption of radiation in eorresponding detection devices (gamma camera, SPECT deviees) relative to a low absorption in the human organism and easy availability by a molybdenum/technetium generator -- is the radionuclide most often used in elinical nuclear medicine. Its short half-life of 6.02 hours guarantees an only slight exposure of the patient to gamma radiation, particularly since also the daughter product technetium-99 has only a negligible residual radiation. But a drawback of the technetium is its complicated and still not completely known complex chemistry. Technetium can be present in a number of oxidation stages (+7 to -1), which can greatly change the pharmacological properties by changing the eharge of a eomplex. It is therefore neeessary to synthesize eomplexes whieh bind the technetium in a defined oxidation stage and to prevent redox reactions, which could lead to a redistribution of the pharmaceutical agent. A number of such Tc-99m complexing agents are already known and are clinically used.
In the case of neutral complexes, systems, in which the Tc-99m is bound between 2-4 nitrogen atoms and 0-2 sulfur atoms (N2S2, N3S
and propylenamine oxime eomplexes), are often involved. But often 2~2~
the insufficient stability of these Tc-99m complexes is a significant drawback (Hung, J. C. et al.; J. Nucl. Med. 29:
1568, 19~8). In clinical use, therefore, e.g., HMPA0 (hexamethyl-propylenamine oxime) has to be administered shortly after its labeling with pertechnetate, so that the portion of decomposition products, which reduce the diagnostic informative value, is not too high. A coupling of these chelates or chelating agents to other substances selectively accumulating in foci of disease is not possible. Therefore, most of said complexes are distributed according to the blood circulation and/or metabolic activity of an organ (e.g., Europ. Patent Appl.
0 194 843), so that, e.g., necrotic or ischemic regions can be represented after infarction or stroke in a scintigram.
But for a successful diagnosis of tumors, neurological diseases or diseases of the cardiovascular system, substances are promising which can produce molecular changes of diseased tissue, by being bound specifically to these diseased tissues or being infiltrated in their metabolism. The findings of the biological and biochemical basic research allow the selection of a number of substances, which selectively accumulate in foci of disease:
various tumors develop increased or reduced surface concentrations on receptors, e.g., for growth factors or steroid hormones (Sledge, G. W.; Adv. Cancer Res. 38: 61-75 [1983]).
Also, in neurological diseases, a change of the concentration of receptors for neutrotransmitters results in specific areas of the brain tFrost, J. J.; Trends Pharmacol. Sci. 7: 490-496 tl987]).
Further, diseased, damaged or cells transformed into tumo-- cells 2 0 ~
often show great changes of their metabolism and an oxygen deficiency inside the tumor. ~he use of such physiological characteristics can be used in in vivo diagnosis, by, e.g., hormones, neurotransmitters or certain metabolic products such as fatty acids, saccharides, peptides or amino acids being coupled to chelating agents for Tc-99m. Also, substances such as misonidazole (a radiosensitizer) or other compounds reacting to radicals in the absence of oxygen can be used for specific accumulation of radioactive isotopes and thus graphic representation of tumors or ischemic regions. Finally, also the coupling to monoclonal antibodies is possible, which bec~use of their high specificity have become a promising instrument in the diagnosis of tumors.
For the production of diagnostic agents according to the described principle, it is necessary that chelating agents for radioactive metal ions, in particular Tc-99m, can be coupled to substances selectively accumulating in diseased tissues.
Since the isotopes of rhenium (Re-188 and Re-186) have chemical properties similar to Tc-99m, the chelating agents can also be used to complex these isotopes. Said Re-isotopes are ~-radiators. Thus, the selectively accumulating substances complexed with rhenium instead of with technetium are also usable in the treatment of tumors.
The previously known attempts to couple chelating agents to selectively accumulating substances in many cases can be considered as unsatisfactory. If the functional groups of the complexing agent are used to bind the chelating agent to such a 2 ~

molecule, a weakening of the complex stability often results, i.e., a diagnostically intolerable portion of the isotope is released from the conjugate. (Brechbiel, M. W. et al., Inorg.
Chem. 25: 2772 [1986])~ It is therefore necessary to produce bifunctional complexing agents, i.e. complexing agents which carry both functional groups for coordinative bonding sf the desired metal ion and a (another) functional group for bonding the selectively accumulating molecule. Such bifunctional ligands make possible a specific, chemically defined bonding of technetium to the most varied biological materials, also then when a so-called prelabeling is performed. Since according to this method, first the labeling with Tc-99m and the isolation of the complexes is performed and this complex is linked only in a second step with a selectively accumulating molecule, the labeled compounds are obtained with a high degree of purity.
Some chelating agents coupled to monoclonal antibodies (e.g., Europ. Patent App. 0 247 866 and 0 188 256) or fatty acids (Europ. Patent Appl. 0 200 492) were described. But as chelating agents, the already mentioned N2S2 systems are used, which are not very suitable because of their low stability. The somewhat more stable N3S chelates showed, coupled to monoclonal antibodies, not so great a loss of Tc-99m from the conjugates (J.
Lister-James; J. Nucl. Med. 30: 793 and Europ. Patent Appl. 0 284 071).
Since both the selectively accumulating substances in their properties and the mechanisms, according to which they are accumulated, are very different, it is further necessary to be able to vary the couplable chelating agents and match the physiological requirements of the coupling participant with respect to lipophilia and hydrophilia, membrane permeability or impermeability, etc.
For these reasons, there exists an urgent need for stable complex compounds, which are coupled or able to couple to various selectively accumulating compounds.
The object of the invention is thus to make available stable chelating agents, which contain a functional group for coupling to a selectively accumulating compound or a selectively accumulating compound coupled with the help of this functional group.
According to the invention, this object is achieved by the compounds of general formula I

Rl R2A
0\~o R3\l I/R3 R4~,N \~

R5 ~ ~ R5 (I) in which Rl stands for hydrogen or a C1_6 alkyl radical optionally substituted with one or two hydroxyl groups, R2 stands for a C1_6 alkylene radical optionally substituted with a hydroxyl group, R3 stands for a hydrogen atom or a Cl_6 alkyl radical, R4 stands for a hydrogen atom, a Cl-6 alkyl radical optionally 2 ~

substituted with a hydroxyl group or for the meaning indicated under RX, R5 stands for a hydrogen atom or a Cl-6 alkyl radical, B stands for a pyrrolyl radical, a substituted phenyl radical of formula II or a substituted pyridine radical of formula III

~Z ~Z

in which Z means a hydroxyl group, amino group or a mercapto group and Y means a hydrogen atom, a carboxy radical or a sulfonyl radical or B stands for a nitrosomethyl radical of formula IV
NOH
~--C~' RX av) in which Rx means a Cl_6-alkyl radical or, together with R4,i~ aLtrimethylene or tetramethylene group to form a 5- or 6-membered ring.
and A stands for an amino radical, a mercapto radical, a hydrazino or carbohydrazido radical, a carboxy radical, a C2_6 alkinyl or C2_6 alkenyl radical, an oxiranyl radical, a fluorinated phenoxycarbonyl radical, a succinimidoxycarbonyl radical optionally substituted with a sodium sulfate radical, an aminophenyl radical or isothiocyanatophenyl radical, 2 ~ `~ 2 1 ~ 1~

and the phenyl group optional:Ly can be substituted in addition by a carboxy radical, a chlorosulfonyl radical or a sulfonic acid radical or contains a compound ~ selectively accumulating in lesions or specific tissues -- optionally bound by a bifunctional linker radical L with the help of said functional group and T stands for monoclonal antibodies or their fragments, hormones, enzymes, ligands for cell membrane receptors, steroids, neurotransmitters, lipids, saccharides, amino acids and oligopeptides, biotin, as well as radiosensitizers, such as, e.g., misonidazole, as well as their complexes with radioactive metal ions --suitable for diagnosis and treatment of tumors -- as well as their salts with inorganic and organic acids or bases.
According to the invention, those compounds according to claim 1 are preferred in which R4 and R5 mean hydrogen atoms or methyl radicals, as well as those compounds according to claim 1, in which A stands for an amino radical, a mercapto radical, a carboxy radical, a C2_6 alkinyl or C2_6 alkenyl radical, an oxiranyl radical, a fluorinated phenoxycarbonyl radical, a succinimidoxycarbonyl radical optionally substituted with a sodium sulfate radical, an aminophenyl or isothiocyanatophenyl radical, and the phenyl group can be substituted by another carboxyl or sulfonic acid radical, and the selectively accumulating compound T optionally contained in A stands for monoclonal antibodies, their fragments, biotin or misonidazole.

2~2 ~ ~
Surprisingly, many of the chelates synthesized and labeled with Tc-99m showed a higher s~ability than the comparable N2S2, N3S and propylenamine oxime chelates. Thus, no decomposition products were able to be observed after 5 hours, e.g., in a substance accordin~ to the invention (~xample 2), which was coupled by an aminophenyl radical to biotin, while this was the case in the comparable HMPAO known in the literature (Hung, J. C.
et al.; J. Nucl. Med. 29: 1568, 1988). Also, by competitive tests, it was possible to be determined that the Tc-99m chelates described in this invention complex better than the comparable N2S2, N3S and propylenamine oxime chelates. The chelates described in the invention are thus definitely better suitable for diagnostic and therapeutic purposes than the previously known chelates.
The production of the compounds according to claim 1 takes place, by substituted malonic acid halides, preferably malonic acid chlorides of general formula IX

Rl ~.2A~
XOC~<COX

in which R1, R2, A' and X have the above-mentioned meanings and functional groups contained in B optionally are present in protected form, being reacted with an amine of general formula X

2 ~ 2 1~. 3 ,'1~

S
IHR3' (VI) in which R3, R4, R~ and B have the above-mentioned meanings, and functiona~ groups contained in B optionally are present in protected form, in an aprotic solvent, preferably dichloromethane, at temperatures of 0-180C, preferably at room temperature, within 2 to 24 hours, preferably 4 hours, optionally by adding suitable bases, e.g., triethylamine, and present protecting groups being cleaved, and in the thus obtained compounds, group A' optionally converted to A -- optionally after protecting the free amino groups and functional groups Z with protective ions, e.g., as a Cu complex -- being generated and then optionally the thus obtained compounds being coupled by this functional group to selectively accumulating compounds T and substituents B
optionally being complexed with the radioactive isotope desired in each case, and protective ions optionally present in the product in advance being removed according to methods known in the literature and the sequence of the steps of complexing with radioactive isotopes and coupling to T being able to be interchanged.
As hydroxy protecting groups, e.g., the benzyl, 4-methoxybenzyl, 4-nitrobenzyl, trityl, diphenylmethyl, trimethylsilyl, dimethyl-t-butylsilyl and diphenyl-t-butylsilyl groups are suitable. In the case of polyols, the hydroxy groups 2~ ,?Ji~
can also be protected in the form of ketals with, e.g., acetone, acetaldehyde, cyclohexanone or benzaldehyde. Further, the hydroxy groups can also be present, e.g., as THP-ether, a-alkoxyethyl ether, ~EM ether or as esters with aromatic or aliphatic carboxylic acids, such as, e.g., acetic acid or benzoic acid. The hydroxy protecting groups can be released according to methods in the literature known to one skilled in the art, e.g., by hydrogenolysis, reductive cleavage with lithium/ammonia, acid treatment of the ethers and ketals or alkali treatment of the esters (see, e.g., "Protective Groups in Organic Synthesis," T.
W. Greene, John Wiley and Sons 1981).
As amino protecting groups, e.g., trifluoroacetyl, t-butoxycarbonyl, 2,2,2-trichloroethoxycarbonyl, benzoxycarbonyl and acetyl groups are suitable. The amino protecting groups can be cleaved according to methods known in the literature, e.g., by basic or acid hydrolysis, reductive cleavage with zinc in acetic acid or hydrogenolysis.
As mercapto protecting groups, Cl_6 alkyl radicals or benzylthioether are suitable. The cleavage of the mercapto protecting groups takes place selectively with alkali alkylthiolates, alkali alcoholates or alkali metals, preferably with sodium methylthiolate in a polar solvent, preferably in HMPT, DMF or dimethylacstamide or by reductive cleavage with sodium/ammonia.
The functional group in radical A is suitable for producing a stable compound to proteins or other selec~ively accumulating molecules. By the corresponding selection of the functional group in radical A, the coupling is possible under gentle reaction conditions, which do not influence the biological function and/or selectivity.
The coupling to the desired compounds also takes place according to methods known in the art (e.g., Fritzberg et al.; J.
Nucl. Med.: 26, 7 [198~]), for example, by reaction of the corresponding functional groups in radica~ A with nucleophilic groups of the selectively accumulating molecule or, if a nucleophile is involved in the corresponding functional group in radical A itself, with activated groups of the selectively accumulating molecule.
The coupling to steroids with a 17-iodine vinyl function can take place by an ethinyl function of the complexing agent. In this case, it is unimportant whether the E- or Z-compound is used. The removal of the optionally present protecting groups can take place both before and after the coupling. According to the invention, 17~-ethinylestradiol and 17a-ethinylnortestosterone are preferred. These compounds have achieved an importance for the diagnosis of steroid-dependent tumors ~J. K. Mazaitis, B. E. Francis, W. C. Eckelmann, R. E.
Gibson, R. C. Reba, J. W. Barnes, G. E. Bentley, P. M. Grant, H.
A. O'Brien, Jr.: J. Labelled Compd. Radiopharm. 1980, 18, 1033].
The production of 17-iodine vinyl steroids can take place according to processes known in the literature te.g., H.
Hofmeister, H. Laurent, P.-E. Schulze and ~. Wiechert, Tetrahedron 42, 3575 (1986)].

The functional group in radical A represents every substituent which on the one hand allo~s a coupling to a selectively accumulating molecule under mild conditions (e.g., by acylation or amidation~ as well as every activated group, which can react with nucleophilic groups of proteins, antibodies, hormones or other biomolecules, such as the amino, phenol, sulfhydryl, formyl or imidazole group. By activated group is to be understood a function which is capable of reacting with the formation of a conjugate with a nucleophilic substituent of a selectively accumulating molecule or of the complex ligand itself in aqueous solution within a suitably short time, under reaction conditions which entail neither denaturation nor loss of the biological activity. Examples in this respect are imide esters, alkylimide esters, amidoalkylimide esters, succinimide esters, acylsuccinimides, phenol esters, substituted phenol esters, tetrafluorophenol esters, anhydrides, and Michael acceptors. The functional group in radical A is preferably an activated ester (such as phenol or imide ester), a mercaptan or isothiocyanate, in particular for the coupling with nucleophilic groups of amino acids or an aliphatic or aromatic primary amine for the coupling to carbohydrate radicals of proteins.
If a nucleophile is involved in the case of the functional group in radical A itself, it can react with activated groups of a selectively accumulating molecule, and groups of the molecule reacted with so-called "crosslinking reagents" are also enclosed, such as homobifunctional imido esters', homobifunctional N-hydroxysuccinimide esters ~NHS) and heterobifunctional 2 ~

"crosslinkers," which contain various functional groups, such as NHS esters, pyridyl disulfides and activated halogens, such as ~-keto halides. Such crosslinkers are commercially available.
As coupling participants (= compound T), i.a., various biomolecules are provided: ligands, which bind to specific receptors and thus can detect a tissue changed in its receptor density; they include, i.a., peptide and steroid hormones and neurotransmitters. With ligands for steroid hormone receptors, the possibility of an improved diagnosis of breast and prostate carcinomas was demonstrated (S. J. Brandes & J. A.
Katzenellenbogen, Nucl. Med. Biol. lS: 53, 1988). Often, ligands labeled with positron-emitting isotopes for neuroreceptors were able to be used for diagnosis of various brain diseases (J. ~. -Forst, Trends in Pharmacol. Sci. 7: 490, 1987). Other biomolecules are metabolites able to infiltrate the metabolism of the cells, which make a changed metabolism recognizable; they include, for example, fatty acids, saccharides, peptides and amino acids. Fatty acids coupled to the more unstable N2S2 chelating agents were described in EPA 0 200 492. Other metabolism products such as saccharides (desoxyglucose), lactate, pyruvate and amino acids (leucine, methylmethionine, glycine) were used with the help of the PET technique for graphic representation of changed metabolic processes (R. Weinreich, Swiss Med. 8, 10, 1986). Also, nonbiological substances such as misonidazole and its derivatives, which in tissues or tissue parts with reduced oxygen concentration bind irreversibly to cell components, can be used for specific accumulation of radioactive 1~
2~?~

isotope~ and thus graphic representation of tumors or ischemic regions (M. ~. Shelton, J. Nucl. Med. 30: 351, 1989). Finally, the coupling of the bifunctional chelating agents to monoclonal antibodies or their fragments is also possible. The compounds according to the invention containing biotin make possible the bonding of the radioactive conjugates to substances containing avidin or streptavidin. This can be used to accumulate antibody- -streptavidin conjugates on the tumor and Gnly later to administer the radioactive biotin-containing components, which results in a reduced exposure of the patient to radioactive radiation (D. J.
Hnatowich et al., J. Nucl. Med. 28: 1294, 1987). By complexing the conjugates with Tc-99m or rhenium isotopes, a diagnosis and treatment of tumors can be made possible.
It is unimportant whether a ]abeling of the chelating agents with Tc-99m or a rhenium isotope is performed before or after the coupling to the selectively accumulating molecule. But for a coupling to the selectively accumulating molecule after a complexing, it is a prerequisite that the reaction of the radioactive complex with the accumulating compound occurs quickly, under gentle conditions and almost quantitatively, and no subsequent purification is necessary.
The production of the pharmaceutical agents according to the invention takes place in a way known in the art, by the complexing agents according to the invention being dissolved while adding a reducing agent, preferably tin (II) salts such as tin chloride or tin tartrate -' and optionally by adding the 1~ 2 ~

additives usual in galenicals -- in aqueous medium and then being sterilized by filtration.
Suitable additives are, for example, physiologically harmless buffers (e.g., tromethamine), small additions of electrolytes (e.g., sodium chloride), stabilizers (e.g., gluconate, phosphates or phosphonates). The pharmaceutical agent according to the invention is present in the form of a solution or in freeze-dried form and is mixed shortly before the administration with a solution of Tc-99m pertechnetate, eluted from commercially available generators, or a perrhenate solution.
In the nuclear medical in vivo use, the agents according to the invention are provided in amounts of 1-10-5 to 5-104 nmol/kg of body weight, preferably in amounts between 1-10-3 and 5-102 nmol/kg of body weight. Starting from an average body weight of 70 kg, the amount of radioactivity for diagnostic uses is between 0.05 and 50 mCi, preferably 5 to 30 mCi per administration. For therapeutic uses, between 5 and 500 mCi, preferably 10-350 mCi is administered. The administration is normally performed by intravenous, intraarterial, peritoneal or intratumoral injection of 0.1 to 2 ml of a solution of the agent according to the invention. The intravenous administration is preferred.
The following examples are used to explain the object of the invention in more detail.

2~2'~
Example 1:
2-Methyl 2-(4-nitrobenzyl)-malonic acid dimethyl ester rl1 11.5 g (0.5 mol) of sodium under a nitrogen stream is added in a dry three-necked flask holding 1000 ml with a reflux condenser and drying tube as well as a dropping funnel with pressure compensation. Then, 350 ml of methanol is carefully instilled and stirred until the sodium is completely dissolved.
In the still warm sodium methanolate solution, the methanolic solution of 87 g (0.5 mol) of methylmalonic acid diethyl ester is instilled very slowly. After completion of the addition, it is stirred for another 30 minutes and then the 4-nitrobenzyl bromide is added in portions with the help of a powder funnel. After all has dissolved, it is first refluxed for 2 hours and then stirred overnight at room temperature. The solvent is drawn off on a rotary evaporator, the residue is mixed with water and extracted several times with ethyl acetate (250 ml each time). The combined organic extracts are washed with saturated common salt solution and dried on sodium sulfate. After removal of the solvent, pale yellow crystals remain. The recrystallization takes place from dimethylformamide (DMF)/water.
Melting point: 94.5 - 95.0C
Yield: 74%
H-NMR data in DMSO/TMS
1.4 ppm (s,3H,Me); 3.5 ppm (s,2H,CH2Ar); 3.8 ppm (s,6H,MeOOC); 7.2 ppm (d,2H,ArH); 8:2 ppm (d,2H,ArH) 2 ~ 4 2-Methyl-2-(4'-nitrobenzyl)-malonic acicl r 2l 28.1 g of 2-methyl-2-(4'-nitrobenzyl)malonic acid ethyl ester (0.1 mol) is dissolved in 100 ml of methanol and a solution of 20 g of NaOH (0.5 mol) in 50 ml of water is slowly instilled in it. Then, it is heated for 2 hours in a water bath to 50C, the solvent is drawn off on a rotary evaporator, and the residue is taken up in water. It is acidified with semiconcentrated hydrochloric acid and the free carboxylic acid is extracted with chloroform, washed and dried.
Yield: 98%
H-NMR data in DMSO/TMS
1.4 ppm (s,3H,Me); 3.1 ppm (d,2H,CH2Ar); 7.4 ppm (d,2H,ArH); 8.2 ppm (d,2H,ArH).

2-Methyl-2-(4~nitrobenzyl)malonic acid dichloride r 31 The mixture of 25.3 g of [2] and 35.7 g of thionyl chloride and one drop of DMF is heated to boiling with exclusion of moisture with stirring. After completion of the gas generation, the excess thionyl chloride is drawn off at room temperature in a vacuum and the residue is immediately further reacted.
Yield: 91%

N N'-bis(2-MethoxYbenzyl)-2-methvl-2-(4~-nitrobenzvl~-malonic acid diamide r 4l The solution of 7.1 g of [3] (24 mmol) in 100 ml of methylene chloride is carefully instilled in the solution of 5.5 g of 2-methoxybenzylamine (40 mmol) and 4.3 g of triethylamine (42 mmol) in 100 ml of methylene chloride with ex~lusion~ ~ 2 moisture. Then, it is stirred for six hours at room temperature and the reaction mixture is washed several times with water, dried, concentrated by evaporation and recrystallized from 2-propanol.
Yield: 78%
H-NMR data in CDC13/TMS
1.3 ppm (s,3H,CH3~; 3.2 ppm (s,2H,CH2Ar); 3.8 ppm (s,6H,CH30Ar); 4.4 ppm (d,4H,ArCH2NH); 6.8-7.4 ppm (m,lOH,ArH);
7.8 ppm (d,2H,ArH) N N'-bis(2-Hydroxvbenzyl)-2-methyl-2-(4'-nitrobenzvl~malonic acid diamide f51 17 g of boron tribromide (70 mmol) in 200 ml of methylene chloride is slowly instilled in a solution of 4.90 mg of t4] (10 mmol) in 150 ml of anhydrous methylene chloride at 0C under an argon atmosphere and then heated to 40C. After 2 hours, it is hydrolyzed with water, al~alized with potassium carbonate solution and extracted with methylene chloride. After drying and removal of the solvent, a pale yellow residue remains, which is recrystallized from 2-propanol.
Yield: 80%
H-NMR data in CDC13/TMS
1.3 ppm (s,3H,CH3); 3.0 ppm (s,2H,CH2Ar); 4.4 ppm (d,4H,ArCH2NH); 6.3 ppm (d,2H,ArH); 6.6 ppm (d,2H,ArH); 6.8 ppm-7.3 ppm (m,6H,ArH); 7.8 ppm (d,2H,ArH); 9.0 ppm (s,2H,NH) ~ ~ ~ 2 c/ ~
N N'-bis(2-Hvdroxybenzvl)-2-(4'-aminobenzvl)-2~methYlmalonic acid diamide r 61 230 mg of 10% Pd/C in 80 ml of MeO~ is suspended in a 250 ml two-necked flask and saturated with hydrogen. Then, the solution of 1.15 g of t5] (2.5 mmol) in 4 ml of methanol is added with a 5 ml one-way syringe by a septum. After completion of the hydrogen absorption, it is separated from the catalyst and the solvent is drawn off in a vacuum. White crystals remain. The recrystallization takes place from methanol.
Yield: 94%
H-NMR data in CDC13/TMS
1.3 ppm (s,3H,CH3); 3.0 ppm (s,2H,CH2Ar~; 4.4 ppm (d,4H,ArCH2NH); 6.3 ppm (d,2H,ArH); 6.6 ppm ~d,2H,ArH); 6.8 ppm-7.3 ppm (m,8H,ArH); 9.0 ppm (s,2H,NH) N N'-bis(2-Hydroxybenzyl)-2-~4'-isothiocvanatobenzvl)-2-methylmalonic acid diamide ~7~
An 85% solution of thiophosgene in carbon tetrachloride (0.2 ml, 2.23 mmol) is added to the solution of 257 mg of [6] (0.5 mmol) in 4 ml of hydrochloric acid (3 M). The reaction mixture is stirred for 3 hours at room temperature and then evaporated to dryness in a vacuum.
Yield: 86%
H-NMR data in CDC13/TMS
1.3 ppm (s,3H,CH3); 3.0 ppm (s,2H,CH2Ar~; 4.3 ppm (d,4H,ArCH2NH); 6.7 ppm-7.3 ppm (m,12H,ArH); 7.7 ppm (t,2H,NH);
8.8 ppm (s,2H,OH) 2~ J'~
Example la N~N~-bis(2-Hydroxybenzyl)-~- r 4'-(biotincarbamoYl)benzYll-2-methylmalonic acid diamide r 81 680 mg of NHS-biotin (2 mmol) is added to a solution of 514 mg of [6] (1 mmol) in 10 ml of DMSO with stirring and heated for 3 hours to 50C. After cooling to room temperature, it is stirred for another 15 hours. Thenl the solvent is drawn off in a vacuum and the residue is purified by MPLC (silica gel, dichloromethane/ethanol/concentrated ammonia 10:20:1).
Yield: 59%
H-NMR data in DMSO/TMS
1.3 ppm (s,3H,CH3); 1.3-1.7 ppm (m,6H,CH2); 2.3 ppm (t,2H,CH2CO); 2.8 ppm (m,2H,CH2S); 3.0 ppm (s,2H,CH2Ar); 3.1 ppm (m,lH,CHS); 4.1 ppm (m,lH,CHNHCO); 4.3 ppm (m,5H,ArCH2NH+CHNHCO);
6.7 ppm-7.3 ppm (m,12H,ArH) Example lb N.N'-bis(2-HYdroxybenzvl)-2-L4'-~4-(N-maleimidomethyl)cyclohexane~carbamovl~benzYll-2-methYlmalonic acid diamide r 91 This compound is produced corresponding to compound [8].
Yield: 45%
H-NMR data in DMSO/TMS
1.3 ppm (s,3H,CH3); 1.1-1.9 ppm (m,9H,CH2); 2.5 ppm (m,lH,CHCO); 3.0 ppm (s,2H,CH2Ar); 3.3 ppm (CHCH2N); 4.4 ppm (d,4H,ArCH2NH); 6.3 ppm (d,2H,ArH); 6.6 ppm (d,2H,ArH); 6.8 ppm-7.3 ppm (m,lOH,ArH+CH=CH).

2~,~2~`~5~
Example lc Tc-99m complex of r6]
A solution of 5.0 mg of [6] in 5 ml of water is filled with ~tOH to 10 ml. 400 microliters of this solution is mixed with 80 microliters of a saturated tin tartrate solution and 8 mCi of Tc-99m pertechnetate from an Mo99/Tc99m generator and allowed to stand for 5 minutes. Then, this solution is extracted three times with 2 ml of chloroform each, the organic phase is dried on a short sodium sulfate column, mixed with 50 microliters of thiophosgene and incubated for 5 minutes at room temperature.
Then, it is mixed with 100 microliters of isopropanol, the chloroform is evaporated with argon and 900 microliters of a 1%
PVP solution in water is added to the residue.

Couplin~ of the Tc-99m complex containinq isothiocYanate to proteins The purified antibody (10 mg/ml~ is treated at pH 3.5 for 2 hours with 25 micrograms/ml of pepsin and then isolated by FPLC.
Before the coupling with the chelating agent, the fraqments are dialyzed at 4C for 12-24 hours from 0.1 M KH2PO4/0.1 M NaHCO3, pH 8.5. The protein concentration is adjusted to 10 mg/ml. The produced Tc-99m complex is added in a molar ratio of 1:10 (complex: protein) to the protein solution. For conjugate formation, the mixture is incubated for one hour at 37C.

2~52/1~
Example 2 N N'-bis r ( 5-Carboxv-2-methoxy~benzyl)l-2-methyl-2-(4 nitrobenzyl)-malonic acid diamide rlOl This compound is produced corresponding to compound [4].
Yield: 56%
H-NMR data in DMS0/TMS
1.3 ppm (s,3H,CH3); 3.2 ppm (s,2H,CH2Ar); 3.8 ppm (s,6H,CH30Ar); 4.5 ppm (d,4H,ArCH2NH); 6.9-8.0 ppm (m,8H,ArH);
8.2 ppm (d,2H,ArH) N~N~-bis~(5~-Carboxv-2'-hydroxy)benzyll-2-(4'-nitrobenzyl)-2 methvlmalonic acid diamidel rlll This compound is produced corresponding to compound [5].
Yield: 79%
H-NMR data in DMS0/TMS
1.3 ppm (s,3H,CH3); 3.1 ppm (s,2H,CH~Ar); 4.4 ppm (d,4H,ArCH2NH); 6.8-8.0 ppm (m,lOH,ArH).

N~N~-bisr(5~-CarboxY-2~-hvdroxy)benzyll-2-(4~-aminobenzyl)-2 methylmalonic_acid diamide r 121 This compound is produced corresponding to compound [6].
Yield: 92%
H-NMR data in DMS0/TMS
1.3 ppm (s,3H,CH3); 3.0 ppm (s,2H,CH2Ar); 4.4 ppm (d,4H,ArCH2NH); 6.8-7.9 ppm tm,lOH,ArH).

2 ~
N, N ' -~is r ( s~-carboxv-2~-hydroxy~benzvll-2-(4~-_sothiocyanatobenzyl)-2-methYlmalonic acid diamide rl31 This compound is produced corresponding to compound [7].

Yield: 83%

H-NMR data in DMS0/TMS

1.3 ppm (s,3H,CH3); 3.0 ppm (s,2H,CH2Ar); 4.4 ppm (d,4H,ArCH2NH); 6.8-7.9 ppm (m,lOH,ArH).

Example 3 _- r 4~-(Tetrahydro-2-pyranvloxv~butvll-malonic acid dimethvl ester rl4l 7.9 g (0.3 mol) of sodium is added in a 500 ml three-necked flask with a reflux condenser and drying tube as well as a dropping funnel with pressure compensation under a nitrogen stream. Then, 250 ml of methanol is carefully instilled anhydrously and stirred until the sodium is completely dissolved.
In the still warm sodium methanolate solution, the methanolic solution of 95.4 g (0.58 mol) of malonic acid diethyl ester is instilled very slowly. After completion of the addition, it is refluxed for another hour and then 0.5 mol of 1-chloro-4-tetrahydro-pyranyloxybutane is slowly instilled. After completion of the addition, it is refluxed overnight. The solvent is drawn off on a rotary evaporator, the residue is mixed with water and extracted several times with ethyl acetate. The combined organic extracts are washed with saturated common salt solution and dried on sodium sulfate. After removal of the 2 ~
solvent, a colorless oil remains. After distillation in a vacuum, 66.4 g of the substituted malonic ester remains.
Yield: 70%
H-NMR data in CDC13/TMS
1.6-2 pp~ (m,12H,CH2); 3.5-3.7 ppm (m,5H,CH~OTHP); 3.7 ppm (s,6H,COOMe); 4.6 ppm (t,lH,OTHP) 2- r 4 ~ - f Tetrahydro-2-pyranyloxv)butvll-malonic acid r 1S 1 This compound is produced corresponding to compound [2].
Yield: 93%
H-NMR data in DMSO/TMS
- 1.3-2.1 ppm (m,12H,CH2); 3.1-4.0 ppm (m,5H,CH,O~HP);
4.5 ppm (t,lH,OTHP).

2- r 4'-(Tetrahydro-2-pYranvloxY3butyll-malonic acid chloride t161 This compound is produced corresponding to compound [3].
Yield: 89%

N,N'-bis r 2-(Nitrobenzvl31-2- r 4'-(tetrahydro-2-pyranvloxv)butvll-malonic acid diamide r 171 - --3.5 g of 2-nitrobenzylamine (23 mmol) in 150 ml of dichloromethane is slowly instilled in a solution of 6.3 g of [16] (12 mmol) and 2.5 g of triethylamine in 100 ml of dichloromethane drop by drop within one hour. The solution is then cooled to 0C, mixed with water and the aqueous phase is extracted several times with dichloromethane and dried on sodium sulfate. After removal of the solvent, a yellow oil remains.

2~2l~3~
Yield: 68%
H-NMR data in CDC13/TMS
1.6-2 ppm (m,12H,CH2); 3.5-3.7 ppm (m,5H,CH+OTHP); 4.5 ppm (s,4H,NHCH2Ar); 4~6 ppm (t,lH,OTHP); 7.7-7.9 ppm (m,6H,ArH);
8.3 ppm (d,2H,ArH) N N'-bisr2-(Nitrobenz~1)1-2-(4-hydroxYbutyl~malonic acid diamide A solution of 26 g of [17] (50 mmol) in 50 ml of methanol is instilled in a solution of 8 ml of sulfuric acid in 150 ml of methanol and stirred for 15 hours at room temperature. It is cooled to 0C, neutralized, extracted with dichloromethane, dried and concentrated hy evaporation.
Yield: 69%
H-NMR data in CDC13/TMS
1.4-1.6 ppm (m,6H,CH2); 3.5 ppm (t,lH,CH); 3.6 ppm (t,2H,CH2OH); 4.5 ppm (s,4H,NCH2Ar); 7.7-7.9 ppm (m,6H,ArH); 8.3 ppm (d,2H,ArH) N~N~-bisr2-tNitrobenzYl)1-2-(4-methanesulfonylbutyl)malonic acid diamide r 19, 500 mg of methanesulfonyl chloride is added to a solution of 1.8 g of [18] (4 mmol) and 606 mg of triethylamine (6 mmol) in 25 ml of dichloromethane at 0C. After 2 hours, the solution is mixed with ice water and extracted several times with dichloromethane, washed and dried. After the concentration by evaporation, a yellow oil remains.

2~, .

2~2`~
Yield: 90%
H-NMR data in CDC13/TMS
1.4-1.6 ppm (m,6H,CH2); 3.0 ppm (s,3H,Me); 3.5 ppm (t,lH,CH); 4.0 ppm (t,2H,CH20R); 4.5 ppm (s,4H,NCH2Ar); 7.7-7.9 ppm (m,6H,ArH); 8.3 ppm (d,2H,Ar~) N N'-bis r 2-tNitrobenzyl)l-2-(4-thiocyanatobutyl)malonic acid diamide r 201 An excess of potassium thiocyanate (100 mmol) is added to a solution of 22.5 g of [19] (43 mmol) in 10~ ml of methanol and refluxed for 36 hours. Then, it is evaporated to dryness and the residue is taken up in chloroform and purified by a flash-ehromatography (ehloroform).
Yield: Ç4%
H-NMR data in CDC13/TMS

1.4-1.6 ppm (m,6H,CH2); 3.1 ppm (t,2H,CH2SCN); 3.5 ppm (t,lH,CH); 4.5 ppm (s,4H,NCH2Ar); 7.7-7.9 ppm (m,6H,ArH); 8.3 ppm (d,2H,ArH) N N'-bisr2-(AminobenzYl)l-2-(4-thioeyanatobutyl)malonie aeid diamide r 211 This eompound is produeed eorresponding to eompound [6].
Yield: 78~
H-NMR data in DMS0/TMS

1.4-1.6 ppm (m,6H,CH2); 3.1 ppm (t,2H,CH2SCN); 3.5 ppm (t,lH,CH); 4.3 ppm (s,4H,NCH2Ar); 6.6-7.6 ppm (m,8H,ArH)]

2 ~
N N'-bisr2-(Aminobenzyl)l-2-~4-mercaPtobutvl)malonic acid diamide L~
A solution o~ 13 g of [20] (28 mmol) in methanol is instilled slowly in a so]ution of 2.7 g of sodium borohydride (70 mmol) in 50 ml of water. ~fter completion of the addition, it is stirred for another 2 hours at 50C, cooled, neutralized with 2N
hydrochloric acid and extracted several times with dichloromethane. After the drying on sodium sulfate and concentration by evaporation, a yellow oil remains.
Yield: ~1%
H-NMR data in CDC13/TMS
1.4-1.6 ppm (m,6H,CH2); 2.4 ppm (t,2H,CH2SH); 3.4 ppm (t,lH,CH); 4.5 ppm (s,4H,NCH2Ar); 6.6-7.6 ppm 9M,8H,ArH) Example 4:
2-t4-Nitrobenzvl~-malonic acid diethyl ester r231 21.4 g of lithium diisopropylamide is added to 210 ml of anhydrous tetrahydrofuran under a nitrogen stream in a dry three-necked flask with a drying tube and dropping funnel with pressure compensation. Then, 58.0 g of malonic acid diethyl ester in 100 ml of anhydrous tetrahydrofuran is instilled at room temperature within 40 minutes. After 30 minutes, the reaction solution is cooled to -62C and 39.1 g of 4-nitrobenzyl bromide in tetrahydrofuran is instilled slowly with vigorous stirring, stirred for another hour at -62C and then the formed precipitate is filtered off cold. The filtrate is concentrated by evaporation on a rotary evaporator, the residue is brought into 2~2~
solution in 300 ml of ethanol at 65C and separated from insoluble residue. After the cooling, 34.7 g of pale yellowish crystals is obtained.
Yield: 65%
H-NMR data in CDCl3/TMS
1-2 ppm (t,6H,CH2CH3); 3.3 ppm (d,2H,CH2Ar); 3.6 ppm (t,lH,CH); 4.1 ppm (q,4H,CH2CH3); 7.4 ppm (d,2H,ArH); 8.1 ppm (d,2H,ArH) 2-(4-Nitrobenzvl)-malonic acid r 24l This compound is produced corresponding to compound [2].
Yield: 98%
H-NMR data in ~MSO/TMS
3.1 ppm (d,2H,CH2Ar); 3.4 ppm (t,lH,CH); 7.1 ppm (s,4H,NH2); 7.4 ppm (d,2H,ArH); 8.2 ppm (d,2H,ArH~

2-(4-Nitrobenzvl)-malonic acid dichloride r251 This compound is produced corresponding to compound t3]
Yield: 76%

N,N'-bisrMethvl-2-pvrrolyll-2-(4'-nitrobenzvl)malonic acid diamide r26l The solution of 6.6 g of [25] (24 mmol) in 100 ml of methylene chloride is carefully instilled in the solution of 3.8 g of 2-aminomethylpyrrole (40 mmol) and 4.3 g of triethylamine (42 mmol) in 100 ml of methylene chloride with exclusion of moisture. Then, it is stirred for six hours at room temperature 2 ~ 3~
and the reaction mixture is washed several times with water, dried, concentrated by evaporation and recrystallized from 2-propanol.
Yield: 76%
H-NMR data in DMSO/TMS
3.1 ppm (d,2H,CH2Ar); 3.4 ppm (t,lH,CH); 3.7 ppm (s,4H,ArCH2NH); 6.1-6.7 ppm (m,6H,ArH); 7.4 ppm (d,2H,ArH); 8.2 ppm (d,2H,ArH) N,N'-bisrMethyl-2-pyrrolyll-2-(4'-aminobenzvl)malonic acid diamide r271 This compound is produced corresponding to compound [6].
Yield: 75%
H-NMR data in DMSO/~MS
3.1 ppm (d,2H,CH2Ar); 3.4 ppm (t,lH,CH); 3.7 ppm (s,4H,ArCH2NH); 6.2-6.9 ppm (m,lOH,ArH) N,N'-bisrMethyl-2-pyrrolvll-2-(4'-isothiocyanatobenzyl~malonic acid diamide r251 This compound is produced corresponding to compound [7].
Yield: 69%
H-NMR data in DMSO/TMS
3.1 ppm (d,2H,CH2Ar); 3.4 ppm (t,lH,CH); 3.7 ppm (s,4H,ArCH2NH); 6.2-7.0 ppm (m,lOH,ArH) 2 ~
Example 5 N N'-~is r ( 2-(Benzylthio)benzyll-2-methYl-2-(4'-nitrobenzyl)malonic acid diamide r29l This compound is produced corresponding to compound [4].
Yield: 72%
H-NMR data in CDCl3/TMS
1.8 ppm (s,3H,Me); 3.1 ppm (d,2H,CH2Ar); 4.1 ppm (s,4H,ArCH2S); 4.3 ppm (s,4H,ArCH2NH); 7.1-7.6 ppm (m,18H,ArH);
8.3 ppm (m,4H,ArH).

N~N'-bis(2-Mercaptobenzyl)-2-methyl-2-(4~-nitrobenzyl)malonic acid diamide r301 3.5 g of sodium is carefully added to a solution, cooled to -SOC, of 5.1 g of [29~ in 50 ml of THF and 100 ml of liquid ammonia and stirred for 4 hours. Excess sodium is carefully destroyed with ammonium chloride and then it is allowed to heat slowly to room temperature. The residue is taken up in water and extracted several times with dichloromethane, the organic phase is dried and concentrated by evaporation.
Yield: 59%
H-NMR data in CDCl3/TMS
1.5 ppm (s,3H,Me); 3.1 ppm (d,2H,CH2Ar); 4.1 ppm (s,4H,ArCH2NH~; 7.0-7.5 ppm (m,lOH,ArH); 802 ppm (d,2H,ArH).

2 ~
N N'-bis(2-Mercaptobenzyl)-2-methyl-2-t4'-aminobenzvl)malonic acid diamide r311 This compound is produced corresponding to compound [6].
Yield: 69%
H-NMR data in CDC13/TMS
1.4 ppm (s,3H,Me); 3.0 ppm (d,2H,CH2Ar); 4.1 ppm ~s,4H,ArCH2NH); 7.0-7.4 ppm (m,lOH,ArH).

N N'-bis(2-Merca~tobenzYl)-2-methYl-2-(4'-i thiocYanatobenzyl)malonic acid diamide r 32~
This compound is produced corresponding to compound [7].
Yield: 49%
H-NMR data in DMSO/TMS
1.4 ppm (s,3H,Me); 3.0 ppm (d,2H,CH2Ar); 4.1 ppm (s,4H,ArCH2NH); 7.0-7.5 ppm (m,10H,ArH).

Example 6 2-Methyl-2-(2-propinyl~-malonic acid dieth~l ester ~331 This compound is produced corresponding to compound [1].
Yield: 66%
H-NMR data in CDC13/TMS
1.5 ppm (s,3H,Me); 2.0 ppm (t,lH,CCH); 2.8 ppm (m,2H,CH2); 3.8 ppm (s,6H,COOMe) 2-Methyl-2-(2-propinYl)-malonic acid r341 This compound is produced corresponding to compound ~2].
Yield: 81%

2 Q ~ ~ ~ r3 ~
H-NMR data in CDC13/TMS
1.4 ppm (s,3H,Me); 2.6 ppm (d,2H,CH2); 2.9 ppm (t,lH,CCH).

2-Methyl-2-(2-propinyl)-malonic acid chloride r351 This compound is produced corresponding to compound [3].
Yield: 91%

N N'-bis r ( 2-Benzylthio)benzvl-2-methYl-2-(2-Propinvl)malonic acid diamide r361 This compound is produced corresponding to compound ~4].
Yield: 67%
H-NMR data in CDC13/TMS
1.5 ppm (s,3H,CH3); 2.0 ppm (t,lH,CCH); 2.8 ppm (d,2H,CH2CC); 4.1 ppm (s,4H,ArCH2S); 4.2 ppm (s,4H,ArCH2NH); 7.1-7.6 ppm (m,18H,ArH).

N N'-bisr(2-Mercaptobenzvl)l-2-methyl-2-t2-proPinyl~malonic acid diamide r371 This compound is produced corresponding to compound [30].
Yield: 70%
H-NMR data in CDC13/TMS
1-5 ppm (s,3H,CH3); 2.0 ppm (t,lH,CCH); 2.8 ppm (d,2H,CH2CC); 4.0 ppm (s,4H,ArCH25); 7.0-7.5 ppm (m,8H,ArH).

~2~
Example 7 N N'-bisr(2-Benzylthio-3-pYridyl~t~ 2-methYl-2-(4'-nitrobenzyl)malonic acid diamide r381 This compound i5 produced corresponding to compound [4].
Yield: 60%
H-NMR data in CDC13/TMS
1.2 ppm (s,3H,Me); 3.5 ppm (s,2H,CH2Ar); 4.3 ppm ts,4H,ArCH2S); 4.4 ppm (s,4H,ArCH2NH); 7.1-7.5 ppm (m,16H,ArH);
8.3-8.5 ppm (m,4H,ArH) N N'-bis r ( 2'-Thio-3'-pYridvl)methyll-2-methyl-2-~4"-nitrobenzyl)malonic acid diamide r391 This compound is produced corresponding to compound [30].
Yield: 61%
H-NMR data in DMSO~TMS
1.2 ppm (s,3H,Me); 3.4 ppm (s,2H,CH2Ar); 4.3 ppm (s,4H,ArCH2NH); 7.1-`7.5 ppm (m,6H,ArH); 8.3-8.5 ppm (m,4H,ArH).

N N'-bis r ( 2~Thio-3~-Pyridyl)methyll-2-methvl-2-(4 aminobenz~l)malonic acid diamide r 401 This compound is produced corresponding to compound [6].
Yield: 66%
H-NMR data in DMS0/TMS
1.2 ppm (s,3H,Me); 3.3 ppm (s,2H,CH2Ar); 4.3 ppm (s,4H,ArCH2NH); 7.1-7.5 ppm (m,8H,ArH); 8.5 ppm (d,2H,ArH).

N,N'-bis r ( 2'-Thio-3'-pvridyl) ethvll-2-methyl-2-(4"- 2 ~ ~ 2 isothiocyanatobenzyl)malonic acid diamide r411 This compound is produced corresponding to compound [7].
Yield: 82%
H-NMR data in DMS0/TMS
1.3 ppm (s,3H,Me); 3.3 ppm (s,2H,CH2Ar); 4.3 ppm (s,4H,ArCH2NH); 7.2-7.5 ppm ~m,8H,ArH); 8.5 ppm (d,2H,ArH).

Example 8 N N'-bis(2-Methoxyb~nzvl)-2-methYl-2-(2-propinvl~malonic acid diamide r 421 This compound is produced corresponding to compound t4].
Yield: 76%
H-NMR data in CDC13/TMS
1.5 ppm (s,3H,CH3); 2.0 ppm (t,lH,CCH); 2.8 ppm (m,2H,CH2); 3.8 ppm (s,6H,CH30Ar); 4.4 ppm (d,4H,ArCH2NH); 6.8-7.4 ppm (m,8H,ArH).

Example 8a 2-Methyl-2-(prop-1-inYl)-1 3-bis-(2-hvdroxY~benzylamidol-propane r43l 200 mg of 2-methyl-2-(prop-1-inyl)-1,3-bis-(2-methoxy-benzylamido)-propane [42] was dissolved in 4 ml of dry dichloromethane. 3.8 ml of a one-molar solution of boron tribromide in dichloromethane was instilled under nitrogen. The mixture was heated for 4 hours to 40C. After cooling, it was hydrolyzed, after separation of the phases and two subsequent 2~fi~

extractions, the solvent was removed and it was chromatographed on silica gel with dichloromethane/acetone (0-30%). A viscous oil was obtained.
Yield: 70%
H-NMR data in CDC13/TMS
1.53 ppm (s,3H,Me); 1.97 ppm (t,lH,CCH); 2.77 ppm (d,2H,CH2-CC); 4.85 ppm (m,4H,ArCH2NCO); 6.80-7.30 ppm (m,8H,ArH); 8.90 ppm (s,2H,OH) Example 9 17B-Hvdroxy-17~-r6.6-bis-(N,N'-~o-methoxv~-benzYl-amino-carbonvl)-hept-l-(E)-en-3-inel-3-methoxv-1.3 5-estratriene r441 4Q mg of (E)-17~-Hydroxy-17a-(2-iodovinyl)-3-methoxy-1,3~5-estratriene and 48 mg of complexing agent [42] were dissolved together with 3.2 mg of bis-(triphenylphosphine)-palladium dichloride and 2.6 mg of copper(I) iodide in 1 ml of toluene (p.a.), mixed with 22 mg of isopropylamine and stirred under nitrogen gassing for 3 hours at 50C. After this time, it was allowed to cool, the solvent was removed and it was purified by liquid chromatography on silica gel with hexane/ethyl acetate (1:1) as mobile solvent.
Yield: 65%
H-NMR data in CDC13/TMS
0.92 ppm (s,3H,Me[18]); 1.20-3.00 ppm (m,lSH,steroid);
1.55 ppm (s,3H,Me); 2.85 ppm (d,2H,CH2-CC); 3.78 ppm (s,3H,steroid-OMe); 3.85 ppm (s,6H,Ar-OMe); 4.45 ppm 36 2~

(m,4H,ArCH2NCO); 5.55 and 6.23 ppm (AB,2H,CH=CH); 6.60-7.35 ppm (m,llH,Ar).

-Hydroxy-17~- r 6,6-bis-(N,N'-~o-hydroxv)-benzvl-amino-carbonyl)-he~t-l-(E)-en-3-inel-3-methoxY-1.3,5-estratriene r451 7.14 mg of (E)-17~-Hydroxy-17~-(2-iodovinyl)-3-methoxy-1,3,5-estratriene and 8 mg of complexing agent [43] were dissolved together with 0.56 m~ of bis-(triphenylphosphine)-palladium dichloride and 0.46 mg of copper(I) iodide in 0.5 ml of toluene (p.a.), mixed with 4 mg of isopropylamine and stirred under nitrogen gassing for 3 hours at 50C. After this time, it was allowed to cool, the solvent was removed and the entire amount was purified on 5 instant HPTLC plates (Merck, silica gel 60, 20 x 20 cm with a concentration zone) with hexane/ethyl acetate (1:1) as mobile solvent.
Yield: 60%
H-NMR data in CDCl3/TMS
0.93 ppm ~s,3H,Metl8]); 1.20-3.00 ppm (m,15H,steroid);
1.55 ppm (s,3H,Me); 2.88 ppm (d,2H,CH2-CC); 3.78 ppm (s,3H,OMe);
4.37 ppm (m,4H,ArCH2NCO); 5.40 and 6.20 ppm (AB,2H,CH=CH); 6.60-7.30 ppm (m,llH,Ar); 8.50 ppm (m,2H,OH) 3.171~-Dihydroxv-17~-r6.6-bis-(N,N'-~o-hydroxy~-benzvl-amino-carbonyl)-heet-l-(E)-en-3-inel-1 3 5-estratriene r46~
8.63 mg of (E~-3,I7~-Dihydroxy-17a-(iodovinyl)-1,3,5-estratriene and 15 mg of complexing agent t43] were dissolved together with 0.7 mg of bis-(triphenylphosphine)-palladium dichloride and 0.58 mg of copper(I) iodide in 0.6 ml of toluene (p.a.), mixed with 5 mg of isopropylamine and stirred under nitrogen gassing for 12 hours at 60C. The solvent was removed and the entire amount was purified on 5 instant HPTLC plates (Merck, silica gel 60, 20 x 20 cm with a concentration zone) with hexane/ethyl acetate (3:7) as mobile solvent.
Yield: 35%
H-NMR data in CDC13/TMS
0.93 ppm (s,3H,Me[18]); 1.20-3.00 ppm (m,15H,steroid);
1.55 ppm (s,3H,Me); 2.88 ppm (d,2H,CH2-CC); 4.36 ppm (m,4H,ArCH2NCO); 5.40 and 6.20 ppm (AB,2H,CH=CH); 6.55-7.25 ppm (m,llH,Ar); 8.50 ppm (m,2H,OH) Example 10 17~-HYdroxv 17-[6 6-bis(N N'-~o-methox~-benz~l-amino-carbon~l)he~t-1-(Z)-en-3-inel-4-estren-3-one r 471 50 mg of (E)-17~-Hydroxy-17-(iodovinyl)-4-estren-3-one and 35 mg of complexing agent [42] were dissolved together with 24 mg of bis-(triphenylphosphine)-palladium dichloride and 12.5 mg of copper(I) iodide in 2 ml of toluene (p.a.), mixed with 23 mg of isopropylamine and stirred under nitrogen for 8 hours at 60C.
After another 60 hours of stirring at room temperature, the solvent was removed and the entire amount was purified on 10 instant HPTLC plates (Merck, silica gel 60, 20 x 20 cm with a concentration zone) with hexane/ethyl acetate (3:7) as mobile solvent.
Yield: 25%

2~2`~

H-NMR data in CDC13/TMS
1.00 ppm (s,3H,Me[18]); 1.20-2.60 ppm (m,20H,steroid);
1.40 ppm (s,3H,Me); 3.23 ppm (AB,2H,CH2-CC); 3.82 ppm (s,6H,OMe);
4.45 ppm (m,4H,ArCH2NC0); 5.83 ppm (s,lH,C4-steroid); 6.34 ppm (AB,2H,CH=CH); 6.80 and 7.20 ppm (m,8H,Ar).

17~-Hydroxv-17~- r 6 6-bis(N,N'-~o-methoxY)-benzvl-amino-c onYl)he~t-l-(E)-en-3-inel-4-estren-3-one r 481 This compound was produced corresponding to compound [47].
Yield: 33%
H-NMR data in CDC13/TMS
1.00 ppm (s,3H,Me[18]); 1.20-2.60 ppm (m,20H,steroid);
1.50 ppm (s,3H,Me); 2.92 ppm (m,2H,CH2-CC); 3.83 ppm (s,6H,OMe3;
4.42 ppm (m,4H,ArCH2NCO); S.82 ppm (s,lH,C4-steroid); 5.45 and 5.95 ppm (AB,2H,CH=CH); 6.85 and 7.20 ppm (m,8H,Ar).

3,3-EthylenedioxY-17 B -hYdroxy-l7~- r 6 6-bistN N'-~o-methoxY~
benzyl-amino-carbonYl~he~t-l-(E)-en-3-inel-5(6)-estrene r4sl This compound was produced corresponding to compound [47].

Yield: 31%

H-NMR data in CDC13/TMS

0.90 ppm (s,3H,Metl8]); 0.85-2.20 ppm (m,20H,steroid);

1.55 ppm (s,3H,Me); 2.92 ppm (m,2H,CH2-CC); 3.85 ppm ~s,6H,OMe);

3-95 ppm (m,4H,OCH2CH2O); 4.45 ppm (m,4H,ArCH2NCO); 5.45 ppm (s,lH,C6-steroid); 5.50 and 6.18 ppm (AB,2H,CH=CH); 6.90 and 7.20 ppm (m,8H,Ar).

2 ~

17~-Hvdroxy-17~-r6,6-bis(N,N'-~o-hydroxy3-benzYl-amin carbonvl)hept-1-(Z~-en-3-inel-4-estren-3-one r 501 This compound was produced corresponding to compound t47]-Yield: 12%
H-NMR data in CDC13/TMS
1.00 ppm (s,3H,Me[18]); 1.20-2.60 ppm (m,20H,steroid);
1.40 ppm (s,3H,Me); 3~26 ppm (AB,2H,CH2-CC); 4.40 ppm (m,4H,ArCH2NCO); 5.81 ppm (s,lH,C4-steroid); 6.30 ppm (AB,2H,CH=CH); 6.80 and 7.20 ppm (m,8H,Ar); 8.50 ppm (m,2H,OH) 3~3-Ethylenedioxy-17~-hydroxy-17a-~6~6-bis(N~N~-~o-hydroxy)-benzyl-amino-carbonyl)hept-1-(E)-en-3-inel-4-estren-3-one r 511 This compound was produced corresponding to compound [47].
Yield: 18 Example 11 Couplinq and labelin~ of a chelatinq aqent containinq isothiocvanate to proteins In the example of F(ab)2 fragments of monoclonal antibody 17-lA, the coupling of Tc-chelating agents containing isothiocyanate of compound [6] to proteins is to be described.
Instead of antibody fragments, any other protein or a substance containing amino groups can be used.
Monoclonal antibody 17-lA is obtained according to methods known in the literature after administration of 107 of the corresponding hybridoma cells in the abdominal space of a BALB/c-mouse and aspiration of the ascitic liquid after 7-10 days. The 2 ~

purification takes place also according to methods known in the literature by precipitation of ammonium sulfate and affinity chromatography by protein A-sepharose. The purified antibody (10 mg/ml) is treated at pH 3.5 for 2 hours with 25 micrograms/ml of pepsin and then isolated by FPLC. Before the coupling with the chelating agent, the fragments are dialyzed at 4C for 12-24 hours from 0.1 M KH2P04/0.1 M NaHC03, pH 8.5. The protein concentration is adjusted to 10 mg/ml. A 5-fold molar excess of the chelating agent containing NCS [example 1] is dissolved in as little as possible of the same buffer and added to the protein solution. For conjugate formation, the mixture is incubated for 3 hours at 37C. Then, the conjugate is dialyzed for 24-4~ hours with repeated buffer change from PBS (phosphate-buffered saline) and the protein concentration is then, if necessary, adjusted again to 10 mg/ml. Until labeling with Tc-99m, the conjugate can be stored after sterilization by filtration at 4C in acid-purified glass vessels.
The labeling of 1 mg of the antibody fragment with Tc-99m coupled with chelating agent [6] takes place by adding 10 mCi of pertechnetate solution (= 1-2 ml) and 100 micrograms of tin(II) chloride in an argon-flushed Na-pyrophosphate solution (1 mg/ml) or by ligand exchange, e.g., by adding the solution to a commercially available glucoheptonate kit mixed with pertechnetate.

2~:2~
Example 12 2-Methyl-2- r ( 3-tert-butyloxycarbonyl)methvll-malonic acid dimethyl ester r 521 The solution of 4.36 g of methylmalonic acid dimethyl ester (0.025 mol) in 5 ml of anhydrous DMF is slowly instilled in a suspension of 600 mg of sodium hydride (0.025 mol) in 25 ml of anhydrous DMF with stirring. After completion of the addition, it is stirred for another 30 minutes, then the solution of 5.85 g of bromoacetic acid-t-butyl ester (0.030 mol) in 10 ml of DMF is instîlled and stirred for another 2 hours at room temperature (TLC control). After complete reaction, it is poured on 300 ml of ice water, neutralized and extracted with a suitable solvent.
After drying on sodium sulfate, the solvent is drawn off and the residue is distilled.
Yield: 70%
H-NMR data in CDC13/TMS
1.4 ppm (s,9H,Me); 1.5 ppm (s,3H,Me); 2.8 ppm (s,2H,CH2C00); 3.8 ppm (t,6H,Me).

2-Methvl-2- r ( 3-tert-butyloxvcarbonvl~methvll-malonic acid r 531 This compound was produced corresponding to compound [2].
Yield: 53%
H-NMR data in DMS0/TMS
1.4 ppm (s,9H,Me); 1.5 ppm (s,3H,Me); 2.6 ppm (s, 2H,CH2COO) .

2 ~ ~3 % `'~

2-Methyl-2-[(3-tert-b~yloxycarbonyl)methyl]-malonic acid dichloride [541 This compound was produced corresponding to compound [3].
Yield: 72~
H-NMR data in CDC13/TMS
1.4 ppm (s,9H,Me); 1.5 ppm (s,3H,Me3; 2.7 ppm (s, 2H,CH2coo) .

N,N'-bis[(2-p-Methoxybenzylthio-3-pyridyl)methvll-2-methvl-2-r2-(tert-butyloxycarbonyl)-methyllmalonic acid diamide r551 This compound was produced corresponding to compound [4].
Yield: 59~
H-NMR data in DMSO/TMS
1.4 ppm (s,9H,Me); 1.5 ppm (s,3H,Me); 2.7 ppm (s,2H,CH2COO), 3.7 ppm (s,6H,OMe); 4.3 ppm (m,8H,ArCH2S,ArCH2NH);
6.8 ppm (d,2H,ArH); 7.1 ppm (dd,2H,ArH); 7.3 ppm (m,4H,ArH); 7.6 ppm (d,2H,ArH~; 8.4 ppm (t,2H,ArH).

N N'-bis r t 2-Mercapto-3-Pvridyl)methyll-2-methyl-2-(2 carboxymethyl)malonic acid diamide r561 580 mg of compound [55] is added to 10 ml of trifluoroacetic acid at room temperature and heated to boiling. After completion of the reaction, the trifluoroacetic acid is drawn off in a vacuum, the residue is taken up in chloroform, washed with water, dried and concentrated by evaporation.
Yield~ 49~
lH-NMR data in DMSO/TMS

2 ~

1.5 ppm (s,3H,Me); 2.6 ppm (s,2H,CH2COO); 4.2 ppm (m,4H,ArCH2NH); 6.8 ppm (d,2H,ArH); 7.1 ppm (dd,2H,ArH); 7.3 ppm ~m,4H,ArH); 7.6 ppm (d,2H,ArH); 8.4 ppm (t,2H,ArH).

Example 12a N~N~-bisf(2-p-Methoxvbenzylthio-3-pyridyl)methyll-2-methyl-2-(2-carboxymethyl)-malonic acid diamide r 571 1.2 g of compound [55] is added to 10 ml of trifluoroacetic acid at -10C and stirred at 0C. After completion of the reaction, the trifluoroacetic acid is drawn off in a vacuum, the residue is taken up in chloroform, washed with water, dried and concentrated by evaporation and chromatographed on silica gelO
Yield: 59%
H-NMR data in DMSO/TMS
1.5 ppm (s,3H,Me); 2.6 ppm (s,2H,CH2COO); 3.7 ppm (s,6H,OMe); 4.3 ppm (m,8H,ArCH2S,ArCH2NH); 6.8 ppm (d,2H,ArH);
7.1 ppm (dd,2H,ArH); 7.3 ppm (m,4H,ArH); 7.6 ppm (d,2H,ArH); 8.4 ppm (t,2H,ArH).

Claims (7)

1. Compounds of general formula I

(I) in which R1 stands for hydrogen or a C1-6 alkyl radical optionally substituted with one or two hydroxyl groups, R2 stands for a C1-6 alkylene radical optionally substituted with a hydroxyl group, R3 stands for a hydrogen atom or a C1-6 alkyl radical, R4 stands for a hydrogen atom, a C1-6 alkyl radical optionally substituted with a hydroxyl group or for the meaning indicated under Rx, R5 stands for a hydrogen atom or a C1-6 alkyl radical, B stands for a pyrrolyl radical, a substituted phenyl radical of formula II or a substituted pyridine radical of formula III
(II) (III) in which Z means a hydroxyl group, amino group or a mercapto group and Y means a hydrogen atom, a carboxy radical or a sulfonyl radical or B stands for a nitrosomethyl radical of formula IV
(IV) in which Rx means a C1-6-alkyl radical or, together with R4 is a trimethylene or tetramethylene group to form a 5- or 6-membered ring.
and A stands for an amino radical, a mercapto radical, a hydrazino radical, a carbohydrazino radical, a carboxy radical, a C2-6 alkinyl or C2-6 alkenyl radical, an oxiranyl radical, a fluorinated phenoxycarbonyl radical, a succinimidoxycarbonyl radical optionally substituted with a sodium sulfate radical, an aminophenyl radical or isothiocyanatophenyl radical, and the phenyl group optionally can be substituted in addition by a carboxy radical, a chlorosulfonyl radical or a sulfonic acid radical or contains a compound T selectively accumulating in lesions or specific tissues -- optionally bound by a bifunctional linker radical L with the help of said functional group, and T stands for monoclonal antibodies or their fragments, hormones, enzymes, ligands for cell membrane receptors, steroids, neurotransmitters, lipids, saccharides, amino acids and oligopeptides, biotin, as well as radiosensitizers, such as, e.g., misonidazole, as well as their complexes with radioactive metal ions --suitable for diagnosis and treatment of tumors -- as well as their salts with inorganic and organic acids or bases.
2. Compounds according to claim 1, wherein radical L
optionally contained in A is developed from bifunctional linkers such as, e.g.,
3. Compounds according to claim 1, wherein T stands for monoclonal antibodies, their fragments, biotin or misonidazole.
4. Metal chelates according to claim 1 with coordinatively bound, radioactive ions of Tc, Re, Cu, Co, Ga, Y and In, preferably Tc and Re.
5. Use of chelates according to claim 4 for in vivo diagnosis and treatment of tumors.
6. Pharmaceutical agents containing at least one chelate according to claim 1, optionally with the additives usual in galenicals.
7. Process for the production of compounds of general formula I

(I) in which R1 stands for hydrogen or a C1-6 alkyl radical optionally substituted with one or two hydroxyl groups, R2 stands for a C1-6 alkylene radical optionally substituted with a hydroxyl group, R3 stands for a hydrogen atom or a C1-6 alkyl radical, R4 stands for a hydrogen atom, a C1-6 alkyl radical optionally substituted with a hydroxyl group or for the meaning indicated under Rx, R5 stands for a hydrogen atom or a C1-6 alkyl radical, B stands for a pyrrolyl radical, a substituted phenyl radical of formula II or a substituted pyridine radical of formula III

(II) (III) in which Z means a hydroxyl group, amino group or a mercapto group and Y means a hydrogen atom, a carboxy radical or a sulfonyl radical or B stands for a nitrosomethyl radical of formula IV

(IV) in which Rx means a C1-6 alkyl radical, which optionally is cyclized together with R4 by a trimethylene or tetramethylene group to a 5- or 6-ring, and A stands for an amino radical, a mercapto radical, a hydrazino radical, a carbohydrazido radical, a carboxy radical, a C2-6 alkinyl or C2-6 alkenyl radical, an oxiranyl radical, a fluorinated phenoxycarbonyl radical, a succinimidoxycarbonyl radical optionally substituted with a sodium sulfate radical, an aminophenyl radical or isothiocyanatophenyl radical, and the phenyl group optionally can be substituted in addition by a carboxy radical, a chlorosulfonyl radical or a sulfonic acid radical or contains a compound T selectively accumulating in lesions or specific tissues -- optionally bound by a bifunctional linker radical L with the help of said functional group, and T stands for monoclonal antibodies or their fragments, hormones, enzymes, ligands for cell membrane receptors, steroids, neurotransmitters, lipids, saccharides, amino acids and oligopeptides, biotin, as well as radiosensitizors, such as, e.g., misonidazole, as well as their complexes with radioactive metal ions --suitable for diagnosis and treatment of tumors -- as well as their salts with inorganic and organic acids, wherein substituted malonic acid halides, preferably malonic acid chlorides of general formula IX

(V) in which R1, R2, A' and X have the above-mentioned meanings and functional groups contained in B optionally are present in protected form, are reacted with an amine of general formula X

(VI) in which R3, R4, R5 and B have the above-mentioned meanings, and functional groups contained in B optionally are present in protected form, in an aprotic solvent, preferably dichloromethane, a temperatures of 0-180°C, preferably at room temperature, within 2 to 24 hours, preferably 4 hours, optionally by adding suitable bases, e.g., triethylamine, and present protecting groups are cleaved and in the thus obtained compounds, group A' optionally converted to A -- optionally after protecting the free amino groups and functional groups Z with protective ions, e.g., as a Cu complex -- are generated and then optionally the thus obtained compounds are coupled by these functional groups to selectively accumulating compounds T and substituents B optionally are complexed with the radioactive isotopes desired in each case, and protective ions optionally in the product in advance are removed according to methods known in the literature and the sequence of steps of complexing with radioactive isotopes and coupling to T
can be interchanged.
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