CA2194561A1 - Technetium-sulphonamide complexes, their use, pharmaceutical agents containing them, and process for producing the complexes and agents - Google Patents

Abstract

The invention relates to novel chelate formers containing sulphonamide groups and their metallic chelates of general formula (I) in which n, m, p, V1, V2, V3, V4, X1, X2, X3, R1, R2, R3 and R4 have different meanings, pharmaceutical agents containing these compounds, their use in X-ray diagnosis and therapy, processes for producing these compounds and agents, and conjugates of these compounds with substances which selectively concentrate themselves in diseased tissue, especially peptides.

Description

- 21 ~4561 Technetium-Sulfonamide Complexes, Their Use, Pharmaceutical Agents Containing the Latter, as well as Process for the Production of the Complexes and Agents The invention relates to the object characterized in the claims, i.e., chelating agents containing new sulfonamide groups, their metal complexes, pharmaceutical agents containing these compounds, their use in radiodiagnosis and radiotherapy, process for the production of these compounds and agents, as well as conjugates of these compounds with substances, especially peptides, selectively accumulating in diseased tissue.
The use of radiopharmaceutical agents for diagnostic and therapeutic purposes has long been known in the area of biological and medical research. In particular, radiopharmaceutical agents are used to visualize certain structures such as, for example, the skeleton, organs or tissues.
The diagnostic application requires the use of such radioactive agents, which, after administration, accumulate specifically in the structures in patients which are to be examined. These locally accumulating radioactive agents can then be traced, plotted or scintiscanned with suitable detectors, such as, for example, scintillation cameras or other suitable imaging processes. The distribution and relative intensity of the detected radioactive agent identifies the point of a structure at which the radioactive agent is present and can display the presence of anomalies in structure and function, pathological changes, etc. In a similar way, radiopharmaceutical agents can be used as therapeutic agents to irradiate specific pathological tissues or areas. Such treatment requires the production of radioactive therapeutic agents, which accumulate in certain structures, tissues or organs. By concentration of these agents, the therapeutic radiation is brought directly to the pathological tissue.
Generally, metallic radionuclides are used as diagnostic agents or therapeutic agents, whereby the metal can be present in free form, as ions or in the form of a metal complex. Examples of metallic radionuclides, which can form complexes, are technetium-99m and various rhenium isotopes. The former is used in diagnosis, the latter in therapy. Besides the metal (complex), in general the radiopharmaceutical agents contain additional suitable vehicles and additives, which allow an injection, inhalation or ingestion by the patient, such as, e.g., physiological buffers, salts, etc.
The radionuclide most often used for nuclear-medicine problems is technetium-99m, which is especially well-suited as radioisotope for in vivo diagnosis because of its advantageous physical properties (no corpuscular radiation, 6 hours of physical half-life, 140 KeV of gamma radiation) and the small radiation exposure following from it. Technetium-99m can be obtained problem-free from nuclide generators as pertechnetate and can be used in this form directly for the produc~ion of kits for routine clinical demand.
The production of radiopharmaceutical agents first requires the synthesis of a suitable ligand. In clinical practice, the 2 1 945~ 1 complex is then produced immediately before use from the respective complexing agent (also named ligand or chelating agent below) and the desired radionuclide ~labeling). In this respect, the complexing agent, which is already present in the form of a freeze-dried kit, is reacted under conditions of complex formation with a solution containing the radionuclide. If, for example, the production of technetium-99m radiopharmaceutical agent is desired, the ligand produced with addition of a suitable reducing agent is mixed with a pertechnetate solution, and the corresponding technetium complex is produced under suitable reaction conditions. These complexes are then administered to the patient in a suitable way by injection, inhalation or ingestion.
The solution containing the radionuclide can, as in the case of technetium-99m, be obtained from a commercially available Mo-99/Tc-99m nuclide generator, or -- as in the case of rhenium-186 -- be obtained directly from a manufacturer. The complex-formation reaction is performed under suitable temperatures (e.g., 20~-100~C) within a few minutes to several hours. To assure a complete complex formation, a larger excess (more than 100 times the excess) of the ligand produced and an amount of reducing agent (e.g., SnCl2, S204, etc.) sufficient for a complete reduction of the radionuclide used are necessary.
Since technetium can be present in a series of oxidation stages (+7 to -1), which can greatly change the pharmacological properties by changes of the charge of a complex, it is necessary to provide chelating agents that can bond the technetium in a stable manner in a defined oxidation stage to prevent an undesirable biodistribution from taking place by redox processes or technetium releases from the corresponding radiodiagnostic agents occurring in vivo, which makes more difficult a reliable diagnosis of corresponding diseases.
The efficiency of radionuclides in in vivo diagnosis and therapy depends on the specificity and the selectivity of the labeled chelates in the target cell. An improvement of these properties can be achieved by coupling chelates to biomolecules according to the "drug-targeting" principle. As biomolecules, antibodies, their fragments, hormones, growth factors and substrates of receptors and enzymes present themselves. Thus, in British Patent Application GB 2,109,407, the use of radioactively-labeled monoclonal antibodies to tumor-associated antigens is described for in vivo tumor diagnosis. Also, direct protein labeling with donor groups (amino, amide, thiol, etc.) of the protein (Rhodes, B. A. et al., J. Nukl. Med. 1986, 27, 685-693) or by the introduction of complexing agents (US 4,479,930 and Fritzberg, A. R. et al., J. Nucl. Med. 1986, 27, 957) with technetium-99m is described. These experimental methods, however, are not available for clinical use, since, on the one hand, the selectivity is too low and, on the other hand, the "background activity" in the organism is too high to make possi~le an in vivo imaging.
As suitable complexing agents for technetium and rhenium isotopes, there are, e.g., cyclic amines, as they are described by Volkert et al. (Appl. Radiol. Isot. 1982, 33; 891) and Troutner et al. (J. Nucl. Med. 1980, 21; 443), but which have the drawback that starting from a pH > 9, they are only able to bond technetium-99m in good yields.
N2O2 systems are present in clinical use, but they are affected by the drawback that the corresponding metal complexes in vivo are not very stable. According to investigations of Pillai and Troutner, the complexes lose up to 30% of complexed metal in the plasma as early as after 1 hour (Pillai, M. R. A., Troutner, D. E. et al.; Inorg. Chem. 1990, 29; 1850).
Noncyclic N4 systems, such as, e.g., the HM-PA0, have their low complex stability as a great drawback. Because of its instability (Ballinger, J. R. et al., Appl. Radiat. Isot. 1991, 42; 315), Billinghurst, M. W. et al., Appl. Radiat Isot. 1991, 42; 607), Tc-99m-HM-PA0 must be administered within 30 minutes after its labeling, so that the portion of decomposition products, which have a different pharmacokinetics and precipitation, can be kept small. Such radiochemical impurities make more difficult the detection of diseases to be diagnosed. A
coupling of these chelates or chelating agents to other substances selectively accumulating in foci of disease cannot be achieved with simple means, so that the latter are generally dispersed unspecifically in the organism.
N2Sz chelating agents (Bormans, G. et al.; Nucl. Med. Biol.
1990, 17; 499), such as, e.g., ethylene dicysteine (EC;
Verbruggen, A. M. et al.; J. Nucl. Med. 1992, 33; 551), meet the requirement of sufficient stability of the corresponding technetium-99m complex, but radiodiagnostic agents with a purity of greater than 69~ form first starting from a pH of the complexing medium > 9.
The previously known N3S systems (Fritzburg, A.; EP 0 173 424 and EP 0 250 013) form stable technetium-99m complexes, but they have to be heated to temperatures of about 100~C to form a homogeneous radiopharmaceutical agent.
In recent years, the need for radiodiagnostic agents accumulating specifically in diseased tissues has risen. This can be achieved if complexing agents can be coupled easily to selectively accumulating substances and in this case not lose their advantageous complexing properties. Since it very often results, however, that after coupling of a complexing agent with use of one of its functional groups to such a molecule, a weakening of the complex stability is observed, the present attempts to couple chelating agents to selectively accumulating substances appear hardly satisfactory, since a diagnostically intolerable portion of the isotope is released in vivo from the conjugate (Brechbiel, M. W. et al; Inorg. Chem. 1986, 25, 2772).
It is therefore necessary to produce bifunctional complexing agents that carry both functional groups for bonding the desired metal ion and a (different, several) functional group for bonding the selectively accumulating molecule. Such bifunctional ligands make possible a specific, chemically defined bond of technetium or rhenium isotopes to the most varied biological materials, even though a so-called pre-labeling is performed.
In EP 0 247 866, EP 0 188 256 and EP 0 200 492, several chelating agents, which are coupled to monoclonal antibodies or fatty acids, are described. As chelating agents, however, the already-mentioned N2S2 systems are used, which are hardly suitable based on their low stability. Since both the selectively accumulating substances in their properties and also the mechanisms, according to which they are concentrated, are very different, it is further necessary to vary the couplable chelating agents and to be able to adapt to the physiological requirements of the coupling partner with respect to its lipophilia, membrane permeability, etc.
The object of the invention is therefore to find complexes or complexing agents which overcome the drawbacks of the prior art, i.e., which -- can be produced at a physiological pH from the corresponding complexing agent and the respective metal oxide/salt, -- can be produced at low temperatures, preferably at room temperature, from the corresponding complexing agent and the respective metal oxide/salt, -- show a high complex stability even under in vivo conditions, -- show a high selectivity or tissue-/organ-specificity.
Furthermore, the complexes must meet the requirements which generally are to be imposed on pharmaceutical agents, such as, e.g., good compatibility (i.e., no side effects), good solubility and complete precipitation.
The object is achieved by this invention.

It has been found that compounds of general formula I

~H2)p V~

~H2)n Rl,~NXl S l ~H2)m R

I ~ 2 R
R

in which V1, V2, V3, *, independently of one another, stand for a carbonyl, >CH(COOH) or -CH2 group, X1 stands for a hydrogen atom, a C1-C12 alkyl radical optionally substituted with a carboxyl, an amino or a thiocyanate group, or a metal ion equivalent of a radioactive metal ion of an element of atomic number 43, 45, 46, 75, 82 or 83, X2, X3, independently of one another, stand for a hydrogen atom or a metal ion equivalent of a radioactive metal ion of an element of the above-mentioned atomic numbers, n, m, p stand for numbers o or 1, whereby m t n - 1 holds true, R1 stands for a hydrogen atom, a carboxyl group or a group --U--Z, in which U stands for a direct bond, a straight-chain or branched, saturated or unsaturated Cl-C20 alkylene radical, which optionally contains a maleimide radical, a succinimide radical, a phenyl radical optionally substituted by 1 to 5 fluorine atoms, an amino or nitro group, one or two imino, phenylene, phenylenoxy, phenylenamino, amide, hydrazide, carbonyl, ureido, thioureido, thioamide, ester group(s), 1 to 2 oxygen, sulfur and/or nitrogen atom(s) as well as optionally 1 to 5 hydroxy, mercapto, oxo, thioxo, carboxy, alkylcarboxylic acid, ester, thiocyanate and/or amino groups, and Z stands for a hydrogen atom, a radical of an amino acid, a peptide, a polynucleotide or a steroid, or a functional group with which optionally the radical of an amino acid, a peptide, a polynucleotide or a steroid is bound, R2 stands for a straight-chain or branched C1-C10 alkyl radical, which optionally contains a -COOH group, a C7-C12 aralkyl.radical or an aromatic compound, which optionally is substituted with a chlorine or bromine atom; a thiocyanate, a methyl, ethyl, carboxyl and/or methoxy group, R4 stands for a hydrogen atom or a carboxyl group or if R1 means a hydrogen atom or a carboxyl group, it stands in addition for a group -U-Z, in which U and Z have the indicated meanings, lo 21 94561 R3 stands for a hydrogen atom, a metal ion equivalent of an element of the above-mentioned atomic numbers; a trifluoroacetate, acetate, benzoate, C1-C6 acyl, a benzoyl, a hydroxyacetyl, an acetamidomethyl radical, a benzoic acid radical optionally substituted with a chlorine or bromine atom, a methyl, ethyl, carboxyl and/or methoxy group, a p-methoxybenzyl radical, an ethoxyethyl radical, an SH protective group, a ~ radical, or if X2, X3 stand for a hydrogen and X1 stands for a hydrogen or an optionally substituted C1-C1z alkyl radical, for a radical of formula II

RVNX1 S~ H2)m ~ R
H 2 )n X ~N
V ~H 2 )p V

in which V1 v2 V3, V4, X1, X2, X3, n, m, p, R1, R and R4 have the indicated meanings, 11 21 ~4561 whereby at least one and at most two radicals V1, V2, V3, V4 stand for a carbonyl group, are very well suited as radiodiagnostic agents and radiotherapeutic agents or for the production of radiodiagnostic agents and radiotherapeutic agents.
The complexing agents (chelating agents) according to the invention, i.e., compounds of general formula I with X1, X2, X3 and R3 in the indicated meanings with the exception of a metal ion equivalent, meet the above-mentioned requirement profile.
They are distinguished more particularly in that they complex the respectively desired metal quickly at the physiological pH and low temperatures. They are therefore especially suitable for routine use in clinical practice.
During the complexing process, monomeric metal complexes of formula I with R3 meaning a metal ion equivalent are from the dimeric chelating agents with R3 meaning a radical of general formula II.
As metal ions, radioactive metal ions of the elements of atomic numbers 43, 45, 46, 75, 82 or 83, such as, e.g., the radioisotopes technetium-99m, rhodium-103, palladium-109, rhenium-186, lead-212 and bismuth-212, are used, whereby the selection of the metal isotope depends on the desired field of use. According to the invention, metal complexes of the elements technetium and rhenium are preferred.
If the complexes of general formula I according to the invention contain isotopes that emit ~y-radiation, such as, e.g., Tc-99m, the latter can be used in single-photon emission tomography (SPECT).

If the complexes of general formula I according to the invention contain isotopes that emit ~-particles, such as, e.g., Bi-211, Bi-212, Bi-213, Bi-214 or B-emitting isotopes, such as, e.g., Re-186 or Re-188, the latter can be used in radiotherapy.
According to the invention, compounds of formula I, in which V1 and V4 each stand for a carbonyl group, v2 and V3 each for a -CH2 group and p for number 0, are preferred.
As radical R1, hydrogen or a carboxylic acid group and especially a group -U-Z in which Z is a hydrogen atom are suitable, but it preferably stands for the radical of a biomolecule with tissue- or structure-specific properties or a functional group optionally present in activated form with which optionally such a biomolecule can be bound.
As examples of biomolecules, there can be mentioned radicals of an amino acid, a peptide, or a steroid, such as, e.g., the known steroid hormones (androgens, gestagens, estrogens, cholesterol, cholic acid derivatives, pregnanes, etc.) as well as polynucleotides such as RNA or DNA.
As examples of functional groups with which optionally the bond of a biomolecule can take place, a -COOH, an -SCN, an -OH, a -Cl or an -NH2 group can be mentioned. Such groups can also be present in their activated form, e.g., as succinimide ester or acid chloride.
U can stand for a direct bond but preferably a straight-chain or branched, saturated or unsaturated C1-C20 alkylene radical, which optionally contains a succinimide radical, a phenyl radical optionally substituted by 1 to 5 fluorine atoms, ..

an amino or a nitro group, one or two imino, phenylene, phenylenoxy, phenylenamino, amide, hydrazide, carbonyl, ureido, thioureido, thioamide, ester group(s), 1 to 2 oxygen, sulfur and/or nitrogen atom(s) as well as optionally 1 to 5 hydroxy, mercapto, oxo, thioxo, carboxy, alkylcarboxylic acid, ester, thiocyanate and/or amino groups.
As examples of U, if Z stands for a biomolecule, there can be mentioned a -CH2-C6H4-O-CH2-C6H4-~ -cH2-c6H4 O CO C15 30 ' 2 O (CH ) COO- -CH2-C6H4-O-cH2-cOO-c6H4 NH ~ 2 6 4 COO-, - (CH2) 5-COO-, or a -CH2-C6H4-O-(CH2) 2-~- (CH2) 2-~-group.
As radicals -U-Z with Z meaning hydrogen, there can be mentioned a -CH2-C6H4-O-CH2-cOO-c6F5~ -CH2-C6H4 ~ CH2 6 5 ~ 2 6 4 3, CH2 C6H4 O-C6H3, -CH2~c6H4~0-c12H2s or a -CH2-C6H4 O CO C15H31 group.
As radicals -U-Z with Z meaning an optionally activated functional group, there can be mentioned a -CH2-(CH2) 4-NCS, -CH2-O ( CH ) -COOH -CH2C6H4-O-CH2-COO-C6H4 NH2 ~ 2 6 4 COO-C6H4-NO2 ~ -CH2-C6H4 NCS, O ~

CH2~30~H.~oo._N~ ---N~3 -CH2-C6H4-O-(CH2)2-O-(CH2)2-OH or a -CH2-C6H4-O-CH2-COOH group, i.e., o Z stands for an -NCS, -COOH, -NH2, -NO2, -OH or a ~oO_N

group.

As examples of radicals RZ, there can be mentioned a -C6H4-NCS or a -C6H4-COOH group, but especially a -CH3 group or a phenyl ring.
As radicals R3, there can be mentioned as examples, a) in the case of complexing agents: -SH protective groups, such as, e.g., a -CO-CH3 , o ~ CO C6H5 ~ CO CF3 or a -C(C6H5) 3 group ~ ~>
or a radical of general formula II, in which V1, V2, V3, V4, X1, X2, X3, n, m, p, R1, R2 and R4 have the indicated meanings, b) in the case of the complexes according to the invention: one of the previously mentioned radicals, with the exception of a radical of general formula II: but in addition, R3 can also stand for a metal ion equivalent of a radioactive metal ion of an element of the above-mentioned atomic numbers.
In the case of complexing agents, X2, X3 stand for a hydrogen atom and X1 stands for a hydrogen atom or an optionally substituted C1-C12 alkyl radical, in the case of complexes, depending on the oxidation stage of the metal in the complex, at least 2 of radicals x1, X2, X3 or R3 have the meaning of a metal ion equivalent.
R4 stands for hydrogen or a carboxyl group, but R4 can also stand for a group -U-Z with U and Z in the previously-indicated meanings, whereby it always holds true that at most one of two radicals R1 or R4 means a group -U-Z. According to the invention, compounds with R4 meaning hydrogen are preferred.
Indices m and n stand for numbers O or l. Since isomeric compounds can accumulate in the synthesis of complexing agents according to the invention, it holds true that the sum of m and n is always 1.
The invention also relates to the process for the production of complexing agents and complexes according to the invention, whereby suitably different reaction paths are followed in the synthesis of the complexing agent as a function of the desired target structure. Some typical syntheses are described as examples below. Other complexing agents can be produced analogously to the synthesis methods described.
1. If ligands are to be produced, which contain an -O-C6H4-CH2 group in R1, a start is advantageously made from a tyrosine ester, whose amino group first is protected in the way known to one skilled in the art by reaction with a reagent Z1-Cl, in which Z1 stands for any amino protective group, preferably for a benzyloxycarbonyl group (also named Z group below). Then, the phenolic hydroxyl group is alkylated with an alkyl iodide in a way known in the art, the amino protective group is cleaved off in acidic form and then tosylated, e.g., with toluenesulfonic acid chloride. The thus obtained intermediate product is reacted in an aminolysis of the ester function with ethylenediamine.
Then, it is reacted with 2-acetylmercaptosuccinic anhydride to the complexing agents according to the invention, whereby both isomers are always obtained in the last reaction step.
2. The production of ligands containing an -o-CO-CH2-O-C6H4-CHz group in R1 according to the invention is carried out by first, as described above, the corresponding Zl-protected compound -- starting from tyrosine ester -- being produced. The phenolic hydroxyl group is then alkylated with bromoacetic acid-t-butyl ester, and the amino protective group is cleaved off in a way known in the art. Tosylation and aminolysis of the ester function with ethylenediamine are carried out as described under 1, then it is reacted, however, with chloroacetyl chloride.
Chlorine is substituted in a way known in the art by reaction with potassium thioacetate or sodium thioacetate, and the t-butyl ester is acidically saponified. Optionally, the resulting carboxylic acid group can be activated with hydroxysuccinimide and then reacted with the respectively desired biomolecule.
3. Ligands according to the invention, in which R1 stands for an SCN-C6H4-CH2 radical, can be obtained by first the amino group of a p-nitrophenylalanine ester being tosylated in a way known in the art. Then, the nitro group is hydrogenated, and the resulting aromatic amino group is protected, e.g., by reaction with benzyl chloroformate. The aminolysis of the ester function with ethylenediamine follows this reaction step, as described in the preceding cases, followed by a reaction with S-protected mercaptoacetic acid. The cleavage of the benzyloxycarbonyl group follows the latter. The introduction of the isothiocyanate group is carried out by reaction with thiophosgene.

4. The production of the ligands according to the invention with x1 meaning an alkyl radical is carried out by first the amino group of a glycine ester being tosylated with toluenesulfonyl chloride. Then, the ester function is reacted in an aminolysis with ethylenediamine. The resulting free amino group is protected with a tert-butoxycarbonyl group (designated below as a BOC group) and the nitrogen substituted with a tosyl radical is N-alkylated with an alkyl iodide in a way known in the art. After the cleavage of the BOC group, the reaction with 2-acetylmercaptosuccinic anhydride follows, whereby again a mixture of isomers is obtained.

5. Ligands according to the invention with R1 meaning an aminobutyl radical can be obtained by the BOC-protected lysine ester on a nitrogen atom first being tosylated on the remaining unprotected primary amino group and then being reacted with ethylenediamine in an aminolysis. Below, it is reacted with 2-acetylmercaptosuccinic anhydride (whereby a mixture of isomers results) and the amino protective group is cleaved off according to known methods.

6. The production of ligands, according to the invention, that contain an isothiocyanatophenyl radical as R2 is carried out by reaction of a glycine methyl ester with nitrobenzenesulfonyl chloride, subsequent reduction of the nitro group and protection of the resulting amino group. The thus obtained intermediate compound is reacted in an aminolysis with ethylenediamine and then with S-protected mercaptoacetic acid. The subsequent cleavage of the (BOC) protective group is carried out in a known way. The free amino group is then reacted with thiophosgene to the corresponding isothiocyanate group.
The production of the metal complexes of general formula I
according to the invention with at least two radicals Xl, x2, X3 and/or R3 in the meaning of a metal ion equivalent is carried out in a way known in the art, by the complexing agents according to the invention (which can be obtained as previously described) being dissolved in aqueous medium with the addition of a reducing agent, preferably tin(II) salts, such as tin chloride or tin tartrate -- and optionally with the addition of the additives usual in galenicals, such as physiologically harmless buffers (e.g., tromethamine), small additions of electrolytes (e.g., sodium chloride), stabilizers (e.g., gluconate, phosphates or phosphonates), etc. -- and then sterilized by filtration. In the case of short-lived metal isotopes, such as, e.g., Tc-99m, this solution is reacted immediately before administration with an aqueous solution of the respectively desired metal ion. In the case of long-lived isotopes, the reaction with the metal salt/oxide can even be performed by the manufacturer of radiopharmaceutical agents. To assure a complete complexing of the metal isotope, the complexing agent is generally added in at least a 100-fold excess, i.e., besides the desired metal complex, the agents according to the invention contain in addition the metal-free complexing agent, which is added advantageously in the form of its potassium salt.
Reaction solutions containing the above-mentioned described metal complex can basically be administered immediately without further working-up.
Since the technetium in particular can be present in a series of oxidation stages (+7 to -1), it is advisable to attach stabilizers to the complexing agents. The latter keep the radionuclide in a stable form, until it has reacted completely with the complexing agent (ligand). The stabilizers, which are ~ .

known as transfer or auxiliary ligands, first complex the metal in an exactly defined oxidation stage and then provide it to the target ligands (complexing agents) in a ligand exchange reaction.
Examples of auxiliary ligands are gluconoheptonic acid, tartaric acid, citric acid (including their salts) or other substances known to one skilled in the art.
Optionally, the obtained metal complexes are mixed with pharmacologically acceptable radiological vehicles. This radiological vehicle should exhibit advantageous properties for the administration of the radiopharmaceutical agent in the form of an injection, inhalation or ingestion. As vehicles there can be mentioned as examples HSA, aqueous buffer solutions, such as, e.g., tris(hydroxymethyl)aminoethane (or their salts), phosphate, citrate, bicarbonate, etc., sterile water, physiological common salt solution, isotonic chloride or dicarbonation solutions or normal plasma ions such as Ca2+, Na+, K+, and Mg2+.
In nuclear-medicine in vivo use, the agents according to the invention are dosed in amounts of 10-5 to 5 x 104 nmol/kg of body weight, preferably in amounts between 10-3 to 5 x 102 nmol/kg of body weight (relative to the metal complex). Starting from an average body weight of 70 kg, the required amount of radioactivity for diagnostic uses is between 1.85 MBq and 1.85 GBq per administration. The administration is carried out normally by intravenous, intraarterial, peritoneal or intratumoral injection of 0.1 to 2 ml of a solution of the agents according to the invention. Intravenous administration is preferred.

2 1 ~456 1 The metal complexes according to the invention with metal ions of the above-mentioned elements and the pharmaceutical agents prepared from them are distinguished by good compatibility and high stability in vivo. The complexing agents according to the invention are distinguished by an easy labeling capability, i.e., they complex the desired metals in high yields at room temperature and neutral pH.
The following examples are used for a more detailed explanation of the object of the invention, without intending that they be limited to this object.

Example 1 a) N-Benzyloxycarbonyl-tyroqine methyl ester A suspension of 97.6 g (0.5 mol) of tyrosine methyl ester in 500-1000 ml of dichloromethane is mixed at room temperature with a solution of 86 g of benzyl chloroformate in 100 ml of dichloromethane. Then, 51 g of triethylamine is slowly added in drops while being cooled, and it is stirred overnight. The mixture is concentrated by evaporation on a rotary evaporator at 40~C, the residue is taken up in 750 ml of EtOAc, and the undissolved material is filtered out. The filtrate is washed three times saturated common salt solution, dried and concentrated by evaporation and recrystallized.
Yield: 72%
Cld: C 65.64~ H 5.82~ N 4.25% O 24.29%
Fnd: C 65.47% H 5.98% N 4.02%

b) N-Benzyloxycarbonyl-2-t~4-hexyloxy)benzyl]-2-aminoacetic acid methyl e~ter 1.12 g of potassium-tert-butylate (10 mmol) is added to a solution of 3.29 g of the compound produced according to Example la) (10 mmol) in 50 ml of DMF at room temperature, and then the solution of 2.12 g of hexyl iodide (10 mmol) in 10 ml of DMF is added in drops and heated for 3 hours to 110~C. After cooling to room temperature, it is poured onto ice, extracted with dichloromethane, washed several times with water, dried, filtered and concentrated by evaporation. After chromatography on a silica gel column (CH2Cl2/EE 19:1), 1.8 g of a pale yellow oil remains.

Yield: 44%
Cld: C 69.71% H 7.56% N 3.39% o 19.35%
Fnd: C 69.54% H 7.69% N 3.13%

c) 2-[(4-Hexyloxy)benzyl]-2-aminoacetic acid methyl ester 4.14 g (10 mmol) of the Z1-protected compound produced according to Example lb) is hydrogenated with hydrogen in 50 ml of ethyl acetate in the presence of 1.5 g of palladium on activated carbon (10%) at 50~C. After the reaction is completed, the catalyst is filtered out, and the solvent is drawn off. 2.7 g of colorless oil remains.
Yield: 97%
Cld: C 68.79% H 9.02% N 5.01% 0 17.18%
Fnd: C 68.64% H 9.11% N 5.09%

d) N-p-Toluenesulfonyl-2t(4-hexyloxy)benzyl]-2-aminoacetic acid methyl ester 8.38 g of the amine (30 mmol) produced according to Example lc) is dissolved in 50 ml of dichloromethane and mixed at 0~C
with 5.72 g of toluenesulfonic acid chloride in 30 ml of dichloromethane. While being stirred intensively, 3.0 g of triethylamine is added in drops at 0~C and stirred for 1 hour at room temperature. After the reaction is completed, it is mixed with ice water and extracted several times with dichloromethane.
The combined organic extracts are washed twice with cold 10% HCl, three times with 10% NaHC03 solution and twice with saturated , NaCl solution. After drying, the solvent is drawn off and chromatographed (silica gel, CH2Cl2).
Yield: 60%
Cld: C 63.72% H 7.21% N 3.23~ O 18.45% S 7.40%
Fnd: C 63.48% H 7.52% N 3.02% S 7.25%

e) N-p-Toluenesulfonyl-2t~4-hexyloxy)benzyl]-2-aminoacetic acidtN-(2-amino-ethyl)]amide A solution of 433 mg of tosyl glycine ester (1 mmol) produced according to Example ld) in 1 ml of anhydrous dichloromethane is slowly added in drops to a solution of 1.2 g of ethylenediamine (20 mmol) in 1 ml of anhydrous dichloromethane and refluxed. After the reaction is completed, it is concentrated by evaporation in a vacuum, and the residue is chromatographed (silica gel, MeOH).
Yield: 42~
Cld: C 62.45% H 7.64~ N 9.10% O 13.864% S 6.95%
Fnd: C 62.36~ H 7.81% N 8.94% S 6.80%

f) N-p-Toluenesulfonyl-2[(4-hexyloxy)benzyl]-2-aminoacetic acid-N-{2-Nt~3-carboxy-2-mercaptoacetyl-1-oxopropyl)]-aminoethyl}amide 0.38 g (2.2 mmol) of 2-acetylmercaptosuccinic anhydride is added to a solution o~ 1.00 g of the amine (2.2 mmol) produced according to Example le) in 10 ml of pyridine/DMF (50:50) and stirred for 4 hours at room temperature. Then, the solvent is drawn off, the residue is taken up in 0.5N HCl and extracted with CH2Cl2. After flash chromatography, 420 mg of a yellow oil remains.
Yield: 31%
Cld: C 56.67~ H 6.50% N 6.61% 0 20.13% S 10.09%
Fnd: C 56.41% H 7.01% N 6.43% S 9.90%

g) Tc-99m complex of N-p-toluenesulfonyl-2[~4-hexyloxy)benzyl]-2-aminoacetic acid-N-{2-Nt~3-carboxy-2-mercaptoacetyl-1-oxopropyl)]aminoethyl}amide 1 mg of the compound produced according to Example lf) is dissolved in 100 ~l of EtOH. 50 ~l of this solution is added to 250 ~l of a 0.1 M phosphate buffer of pH 8.5 and then mixed with 100 ~l of a 99m-Tc-gluconate solution and allowed to stand for 15 minutes. The labeling yield is determined with the aid of HPLC.

Example 2 a) N-Benzyloxycarbonyl-2-[(4-methyloxy)benzyl]-2-aminoacetic acid methyl ester 1.12 g of potassium-tert-butylate (10 mmol) is added to a solution of 3.29 g of the alcohol (10 mmol) produced according to Example la) in 50 ml of DMF at room temperature and then the solution of 1.42 g of methyl iodide (10 mmol) in 10 ml of DMF is added in drops and heated to 110~C. After the reaction is completed, it is allowed to cool to room temperature, poured onto ice, extracted with dichloromethane, washed several times with water, dried, filtered and concentrated by evaporation. After column chromatography (silica gel, CH2Cl2/EE 19:1), 1.92 g of an oil remains.
Yield: 56%
Cld: C 66.46~ H 6.16% N 4.08% 0 23.30%
Fnd: C 66.18% H 6.41% N 3.97%

b) 2-t(4-Nethyloxy)benzyl]-2-aminoacetic acid methyl ester 3.43 g (10 mmol) of the Z1-protected compound produced according to Example 2a) is hydrogenated with hydrogen in 50 ml of ethyl acetate in the presence of 1.5 g of palladium on activated carbon (10%) at room temperature. After the reaction is completed, the catalyst is filtered out, and the solvent is drawn off.
Yield: 98%
Cld: C 63.14% H 7.23% N 6.69% 0 22.94%
Fnd: C 62.91% H 7.36% N 6.50%

c) N-p-Toluenesulfonyl-2t(4-methyloxy)benzyl]-2-aminoacetic acid methyl ester 6.28 g of the amine (30 mmol) produced according to Example 2b) is dissolved in 50 ml of dichloromethane and mixed at 0~C
with 5.72 g of toluenesulfonic acid chloride in 30 ml of dichloromethane. While being stirred intensively, 3.0 g of triethylamine is added in drops at 0~C and stirred for 1 hour at room temperature. After the reaction is completed, it is mixed with ice water and extracted several times with dichloromethane.

The combined organic extracts are washed twice with cold 10% HCl, three times with 10% NaHCO3 solution and twice with saturated NaCl solution. After drying, the solvent is drawn off.
Yield: 76%
Cld: C 59.49% H 5.82% N 3.85% O 22.01% S 8.82%
Fnd: C 59.32% H 6.03% N 3.66% S 8.64%

d) N-p-Toluenesulfonyl-2t(4-methyloxy)benzyl]-2-aminoacetic acid-~N-~2-aminoethyl)]amide A solution of 363 mg of the ester (1 mmol) produced according to Example 2c) in 1 ml of anhydrous dichloromethane is slowly added in drops to a solution of 600 mg of ethylenediamine (10 mmol) in 1 ml of anhydrous dichloromethane and refluxed.
After the reaction is completed, it is concentrated by evaporation in a vacuum, mixed with ethanolic HCl and stirred for 2 hours at room temperature. The solvent is drawn off, and the residue is recrystallized.
Yield: 49%
Cld: C 58.29% H 6.44% N 10.73% O 16.35% S 8.19%
Fnd: C 57.89% H 6.84% N 10.01% S 7.52%

e) N-p-Toluenesulfonyl-2t(4-methyloxy)benzyl]-2-aminoacetic acid-N-{2-N[~3-carboxy-2-mercaptoacetyl-l-oxopropyl)]aminoethyl}amide 0.38 g (2.2 mmol) of 2-acetylmercaptosuccinic anhydride is added to a solution of 941 mg of the amine (2.2 mmol) produced according to Example 2d) in 10 ml of pyridine/DMF (50:50), and it is stirred for 4 hours at room temperature. Then, the solvent is .

drawn off, the residue is taken up in 0.5N HCl and extracted with CH2Cl2. After flash chromatography, 905 mg of a yellow oil remains .
Yield: 73%
Cld: C 53.08% H 5.52% N 7.43% O 22.63% S 11.34%
Fnd: C 52.31% H 5.45% N 7.06% S 11.61%

f) Tc-99m complex of N-p-toluene~ulfonyl-2t~4-methyloxy)benzyl]-2-aminoacetic acid-N-{2-N[(3-carboxy-2-mercaptoacetyl-l-oxopropyl)]aminoethyl}amide 1 mg of the compound produced according to Example 2e) is dissolved in 100 ~1 of EtOH. 50 ~1 of this solution is diluted with 250 ~1 of EtOH, mixed with 50 ~1 of a 0.1 M phosphate buffer of pH 8.5 and then mixed with 100 ~1 of a 99m-Tc-gluconate solution and allowed to stand for 15 minutes. After filtration, the labeling yield is determined with the aid of HPLC.

Example 3 a) N-Benzyloxycarbonyl-2-~(4-dodecyloxy)benzyl]-2-aminoacetic acid methyl ester 1.12 g of potassium-tert-butylate (10 mmol) is added to a solution of 3.29 g of the alcohol (10 mmol) produced according to Example la) in 50 ml of DMF at room temperature, and then the solution of 3.55 g of dodecyl iodide (12 mmol) in 10 ml of DMF is added in drops and heated for 3 hours to 110~C. Then, it is allowed to cool to room temperature, poured onto ice, extracted with dichloromethane, washed several times with water, dried, filtered and concentrated by evaporation. After SC tcolumn chromatography] (silica gel, CHzCl2/EE 19:1), 2.2 g of the desired substance remains.
Yield: 44%
Cld: C 72.40% H 8.71% N 2.81% 0 16.07%
Fnd: C 72.08% H 8.85~ N 2.68%

b) 2-[(4-Dodecyloxy)benzyl]-2-aminoacetic acid methyl ester 4.98 g (10 mmol) of the Z1-protected compound produced according to Example 3a) is hydrogenated with hydrogen in 50 ml of ethyl acetate in the presence of 1.5 g of palladium on activated carbon (10%) at 50~C. After the reaction is completed (4 hours), the catalyst is filtered out, and the solvent is drawn off.
Yield: 91%
Cld: C 72.69% H 10.26% N 3.85% 0 13.20 Fnd: C 72.53% H 9.98% N 3.82%

c) N-p-Toluenesulfonyl-2[l4-dodecyloxy)benzyl]-2-aminoacetic acid methyl e~ter 10.91 g of the amine (30 mmol) produced according to Example 3b) is dissolved in 50 ml of dichloromethane and mixed at 0~C
with 5.72 g of toluenesulfonic acid chloride in 30 ml of dichloromethane. While being stirred intensively, 3.0 g of triethylamine is added in drops at 0~C and stirred for l hour at room temperature. After the reaction is completed, it is mixed with ice water and extracted several times with dichloromethane.

The combined organic extracts are washed twice with cold 10% HCl, three times with 10% NaHC03 solution and twice with saturated NaCl solution. After drying, the solvent is drawn off and chromatographed (silica gel, CHzCl2).
Yield: 75%
Cld: C 67.2B% H 8.37% N 2.71% 0 15.45% S 6.19%
Fnd: C 66.96% H 8.26% N 2.60% S 6.11%

d) N-p-Toluenesulfonyl-2t~4-dodecyloxy)benzyl]-2-aminoacetic acidtN-(2-aminoethyl)]amide A solution of 3.3 g of the ester (6.38 mmol) produced according to Example 3c) in 10 ml of anhydrous dichloromethane is slowly added in drops-to a solution of 1.92 g of ethylenediamine (31.9 mmol) in 20 ml of anhydrous dichloromethane and refluxed for 4 hours. After the reaction is completed, it is concentrated by evaporation in a vacuum, and the residue is recrystallized.
2.3 g of white crystals remains.
Yield: 80%
Cld: C 66.02% H 8.68% N 7.70% 0 11.73% S 5.86%
Fnd: C 65.88% H 8.80% N 7.59% S 5.76%

e) N-p-Toluenesulfonyl-2[t4-dodecyloxy)benzyl]-2-aminoacetic acid-N-{2-Nt(3-carboxy-2-mercaptoacetyl-1-oxopropyl)~-aminoethyl}amide 0.32 g (1.8 mmol) of 2-acetylmercaptosuccinic anhydride is added to a solution of 1.00 g of the amine (1.8 mmol) produced according to Example 3d) in 10 ml of pyridine/DMF (50:50), and it .,.

is stirred for 4 hours at room temperature. Then, the solvent is drawn off, the residue is taken up in 0.5N HCl and extracted with CH2Cl2. After flash chromatography (silica gel, EtOAc/MeOH 9:1), 420 mg of a yellow oil remains.
Yield: 32%
Cld: C 60.06% H 7.42% N 5.84~ O 17.78% S 8.91%
Fnd: C 59.76% H 7.61% N 5.77% S 8.78%

f) Tc-99m complex of N-p-toluene~ulfonyl-2t(4-dodecyloxy)benzyl~-2-aminoacetic acid-N-{2-Nt(3-carboxy-2-mercaptoacetyl-l-oxopropyl)]aminoethyl}amide 1 mg of the compound produced according to Example 3e) is dissolved in 100 ~1 of EtOH. 50 ~1 of this solution is added to 250 ~1 of a 0.1 M phosphate buffer of pH 8.5 and then mixed with 100 ~1 of a 99m-Tc-gluconate solution and allowed to stand for 15 minutes. The labeling yield is determined with the aid of HPLC.

Example 4 a) N-p-Toluenesulfonyl-2[(4-benzyloxy)benzyl]-2-aminoacetic acid-N-{2-Nt~3-carboxy-2-mercaptoacetyl-1-oxopropyl)]aminoethyl}amide The solution of 4.68 g (10 mmol) of the amino compound produced according to Example 16b) in 150 ml of THF is slowly added in drops to a solution of 3.48 g of acetylmercaptosuccinic anhydride (20 mmol) in 50 ml of THF at room temperature and stirred overnight. Then, it is allowed to crystallize out at -20~C. 2.8 g of white crystals remains.

r Yield: 44%
Cld: C 58.02% H 5.50% N 6.55% O 19.95% S 9.99%
Fnd: C 57.80% H 5.72% N 6.51% S 9.85%

b) Tc-99m complex of N-p-toluenesulfonyl-2t~4-benzyloxy)benzyl]-2-aminoacetic acid-N-{2-N[~3-carboxy-2-mercaptoacetyl-1-oxopropyl)]aminoethyl}amide 1 mg produced according to Example 4a) is dissolved in 100 ~l of EtOH. 50 ~l of this solution is added to 250 ~l of a 0.1 M
phosphate buffer of pH 8.5 and then mixed with 100 ~1 of a 99m-Tc-gluconate solution and allowed to stand for 15 minutes. The labeling yield is determined with the aid of HPLC.

Example 5 a) N-p-Toluenesulfonyl-~4-nitrophenyl)alanine methyl e~ter 7.56 g of 4-nitrophenylalanine methyl ester (30 mmol) is suspended in 30 ml of water and mixed at 0~C with 5.72 g of toluenesulfonic acid chloride in 20 ml of diethyl ether. While being stirred intensively, 3.0 g of anhydrous sodium carbonate is added in portions at 0~C within one hour, and it is stirred overnight at room temperature. It is mixed with water and extracted several times with ethyl acetate. The combined organic extracts are washed twice with cold 10% HCl, three times with 10%
NaHCO3 solution and twice with saturated NaCl solution. After drying, the solvent is drawn off. 8.6 g of yellowish crystals remains.

Yield: 76%

Cld: C 53.96% H 4.80% N 7.40% O 25.37% S 8.47%
Fnd: C 53.79% H 4.99% N 7.29% S 8.30%

b) N-p-Toluenesulfonyl-~4-aminophenyl)alanine methyl ester The solution of 2.0 g of N-p-toluenesulfonyl-(4-nitrophenyl)alanine methyl ester in methanol is added to the suspension of 500 mg of Pd/C (10%) in 25 ml of methanol, and it is hydrogenated at 55~C with hydrogen. After filtration and concentration by evaporation, 1.4 g of whitish crystals remains.
Yield: 76%
Cld: C 58.60% H 5.79% N 8.04% O 18.37% S 9.20%
Fnd: C 58.09% H 5.99% N 8.80% S 9.01%

c) N-p-Toluenesulfonyl-{4-tN-(tert-butyloxycarbonyl)laminophenyl}-alanine methyl ester A solution of 3.78 g (10 mmol) of the amino compound produced according to Example 5b) and 3 g of triethylamine (30 mmol) in 50 ml of dioxane is mixed in one portion with 7.4 g of di-tert-butyl-dicarbonate (34 mmol). (Gas generation!) The solution is stirred for 3 hours at room temperature, then poured onto ice water and extracted three times with ethyl acetate. The combined organic phases are washed three times with water and once with saturated common salt solution, dried (MgSO4), concentrated by evaporation and recrystallized.
Yield: 89%
Cld: C 58.91% H 6.29% N 6.25% O 21.40 S 7.15%
Fnd: C 58.12% H 6.69% N 6.21% S 6.59%

d) N-p-Toluenesulfonyl-{4-tN-(tert-butyloxycarbonyl)]aminophenyl}-alanine[N-~2-amino-ethyl)]amide The solution of 700 mg of N-p-toluenesulfonyl-{4-[N-(tert-butyloxycarbonyl)]aminophenyl}-alanine methyl ester (1.6 mmol) in 1 ml of anhydrous dichloromethane is added in drops to a solution of 450 mg of ethylenediamine (7.5 mmol) in 5 ml of anhydrous dichloromethane and refluxed for 8 hours. After the reaction is completed, it is concentrated by evaporation in a vacuum, and the residue is chromatographed (silica gel, ethyl acetate/MeOH 9:1).
Yield: 85%
Cld: C 57.97% H 6.77% N 11.76% O 16.79% S 6.73%
Fnd: C 57.08% H 6.12% N 12.33% S 6.41%

e) N-p-Toluene~ulfonyl-{4-lN-~tert-butyloxycarbonyl)]aminophenyl}-alaninetN-(chloroacetylaminoethyl)]amide The solution of 1.24 g of chloroacetyl chloride (11 mmol) in 5 ml of dichloromethane is added in drops to a solution of 4.77 g of the amine (10 mmol) produced according to Example 5d in 10 ml of dichloromethane at 0~C, then the solution of 2.02 g of triethylamine in 5 ml of dichloromethane is slowly added in drops (attention: strong heat build-up) and stirred overnight. Then, it is mixed with water, extracted with EtOAc, washed neutral with water, dried and concentrated by evaporation. 2.77 g of white crystals remains.
Yield: 50%

Cld: C 54.29% H 6.01% N 10.13% O 17.38% S 5.80~
Fnd: C 53.76% H 5.78% N 9.66% S 5.92%

f) N-p-Toluene~ulfonyl-{4-~N-~tert-butyloxycarbonyl)]aminophenyl}-alanine~N-~2-~acetylmercapto)acetylaminoethyl)]amide In 5 ml of anhydrous DMF under nitrogen atmosphere, a solution of 238 mg of potassium thioacetate (2 mmol) in anhydrous DMF is added in drops to a solution of 553 mg of the derivative (1 mmol) produced according to Example 5e and a catalytic amount of Nal and heated for 3 hours to 110~C. The hot solution is allowed to cool to room temperature, and it is poured into lN HCl (50 ml). Then, it is extracted with EtOAc, washed neutral with water, dried, concentrated by evaporation and chromatographed (silica gel, EtOAc/MeOH 9:1).
Yield: 35~
Cld: C 54.71% H 6.12% N 9.45% O 18.90~ S 10.82%
Fnd: C 54.40% H 6.45% N 9.25% S 10.04%

g) N-p-Toluenesulfonyl-{4-aminophenyl}-alanine-[N-(2-(acetylmercapto)acetylaminoethyl)]amide, hydrochloride 85 mg of N-p-toluenesulfonyl-{4-[N-(tert-butyloxycarbonyl)]aminophenyl}-alanine[N-(2-(acetylmercapto)acetylaminoethyl)]amide is dissolved in 2 ml of 3 M HCl in ethyl acetate and stirred for 6 hours at room temperature. After the solvent is drawn off, 70 mg of white crystals remains.

, Yield: 100%
Cld: C 49.95% H 5.53%N 10.59% 0 15.12% S 12.12%
Fnd: C 49.32% H 5.76%N 10.20% S 11.77%

h) N-p-Toluene~ulfonyl-{4-isothiocyanatophenyl}-alanine-tN-~2-(acetylmercapto) a cetylaminoethyl)]amide The solution of 11.5 mg of thiophosgene in a little dichloromethane is added in drops with exclusion of moisture to a solution of 53 mg of N-p-toluenesulfonyl-{4-aminophenyl}-alanine-[N-(2-(acetylmercapto)acetylaminoethyl)]amide, hydrochloride and 20 ~l of triethylamine in 4 ml of dichloromethane, and it is stirred for 4 hours at room temperature. After the solvent is drawn off, 45 mg of yellowish crystals remains.
Yield: 84%
Cld: C 51.67% H 4.90% N 10.48% O 14.96% S 17.99%
Fnd: C 50.99% H 5.15% N 10.21% S 18.56%

Example 6 a) N-p-Toluene~ulfonyl-{4-tN-~tert-butyloxycarbonyl)]aminophenyl}-alanine-tN-~2-(benzoylmercapto)acetylaminoethyl)]amide 2 mmol of S-benzoyl-2-mercaptoacetic acid (394 mg) and 4 mmol of NEt3 (560 ~l) and 2 mmol of N-p-toluenesulfonyl-{4-[N-(tert-butyloxycarbonyl)]aminophenyl}-alanine-tN-(2-aminoethyl)]amide (953 mg) are mixed with 5 ml of dichloromethane and cooled to 10~C. Then, 2 mmol (509 mg) of 1-benzotriazolyloxy-tris-(dimethylamino)-phosphonium-hexafluorophosphate-chloride (designated below as BOP-Cl) is added and stirred for 4 hours while being cooled in water, then it is mixed with water and brought to pH 1-1.5 with 4N HCl.
Then, it is extracted with dichloromethane, the combined organic extracts are washed with NaHCO3 and water, dried, concentrated by evaporation and chromatographed.
Yield: 60%
Cld: C 58.70% H 5.85% N 8.56% O 17.10% S 9.79%
Fnd: C 58.04% H 6.03% N 8.39% S 9.30%

b) N-p-Toluenesulfonyl-t4-aminophenyl}-alanine-~N-~2-~benzoyl-mercapto)acetyl-aminoethyl)]amide, hydrochloride A freshly produced solution of 3 M HCl in EtOAc (10 ml, 30 mmol) is added to a solution of 654 mg of the protected amine produced according to Example 6a) (1 mmol) in S ml of EtOAc. It is stirred for 30 minutes at room temperature. The solvent is drawn off, and the residue is dried in a vacuum at room temperature.
Yield: 98%
Cld: C 54.86 H 5.29% N 9.48% O 13.53% S 10.85%
Fnd: C 53.90% H 5.52% N 9.09% S 9.97%

c) N-p-Toluenesulfonyl-t4-isothiocyanatophenyl}-alanine-~N-~2 (benzoylmercapto)-acetylaminoethyl)]amide The solution of 115 mg of thiophosgene in a little dichloromethane is added in drops with exclusion of moisture to a solution of 296 mg of the title compound of Example 6b) (0.5 mmol) and 200 ~1 of triethylamine in 4 ml of dichloromethane, and it is stirred for 4 hours at room temperature. After the solvent is drawn off, a residue remains, which is taken up in chloroform and washed with 0.1% citric acid, washed twice with NaHC03 solution and once with water. After drying and concentration by evaporation, 45 mg of yellowish crystals remains.
Yield: 85%
Cld: C 56.36 H 4.73% N 9.39% 0 13.14% S 16.12%
Fnd: C 56.31% H 4.98% N 9.08% S 17.01%

Example 7 a) N-(3-Nitrobenzenesulfonyl)glycine methyl ester 25.11 g of glycine methyl ester (0.2 mol) is dissolved in 500 ml of dichloromethane and mixed drop by drop with 44.3Z g of 3-nitrobenzenesulfonic acid chloride in 100 ml of dichloromethane at 0~C. While being stirred intensively, 40 g of triethylamine, 50 ml of dichloromethane are added in drops at 0~C and stirred for 1 hour at room temperature. After the reaction is completed (TLC control), it is mixed with ice water and extracted several times with dichloromethane. The combined organic extracts are washed twice with cold 10% HCl, three times with 10% NaHC03 solution and twice with saturated NaCl solution. After drying, the solvent is drawn off.
Yield: 82%
Cld: C 39.42% H 3.68% N 10.22% O 35.00% S 11.69%
Fnd: C 40.31% H 3.37% N 9.89% S 11.04%

b) N-(3-Aminobenzenesulfonyl)glycine methyl ester 2.74 g (10 mmol) of the nitro compound produced according to Example 7a) is hydrogenated with hydrogen in 50 ml of glacial acetic acid in the presence of 1.5 g of palladium on activated carbon (10%) at room temperature. After the reaction is completed, the catalyst is filtered out, and the solvent is drawn off.
Yield: 90%
Cld: C 44.26% H 4.95% N 11.47% 0 26.20% S 13.13%
Fnd: C 44.01% H 5.11% N 12.08% S 12.76%

c) N-t~3-Aminobenzenesulfonyl)]glycyl-tN~-(2-aminoethyl)]amide - A solution of 1 g of the N-(3-aminobenzenesulfonyl)-glycine methyl ester (3.6 mmol) in 2.5 ml of anhydrous dichloromethane is slowly added in drops, refluxed, to a solution of 2.2 g of ethylenediamine (36 mmol) in 2.5 ml of anhydrous dichloromethane.
After the reaction is completed, it is concentrated by evaporation in a vacuum, and the residue is chromatographed (silica gel, CH2Cl2/MeOH 9:1).
Yield: 76%
Cld: C 44.11% H 5.92% N 20.57% O 17.63% S 11.78%
Fnd: C 43.09% H 6.41% N 21.82% S 10.68%

d) N-~(3-Aminobenzenesulfonyl)~glycyl-{N'-[~acetylmercapto)acetyl~2-aminoethyl)}amide 250 mg (1.08 mmol) of SATA (SIGMA Chemie [SIGMA Chemistry], 1994; A 9043) in THF is slowly added in drops under argon and , with exclusion of moisture to a solution of 272 mg of amine (1 mmol) produced according to Example 7c) in anhydrous THF at 0~C, and it is stirred for 2 hours at 0~C, then for 12 hours at room temperature. The solvent is drawn off, and the residue is chromatographed (silica gel, CH2Cl2/MeOH 9:1).
Yield: 60%
Cld: C 43.29% H 5.19% N 14.42% O 20.59% S 16.51%
Fnd: C 42.45% H 5.23% N 15.64% S 15.36%

e) N-t(3-Isothiocyanatobenzenesulfonyl)]glycyl-{N'-1(acetylmercapto)acetyll-~2-aminoethyl)}amide The solution of 65 mg of thiophosgene in a little dichloromethane is added in drops to a solution of 200 mg of N-[(3-aminobenzenesulfonyl)]glycyl-{N'-[(acetylmercapto)-acetyl]-(2-aminoethyl)}amide and 20 ~l of triethylamine in 4 ml of dichloromethane with exclusion of moisture, and it is stirred for 4 hours at room temperature. After the solvent is drawn off, a residue remains, which is taken up in chloroform and washed with 0.1% citric acid, washed twice with NaHCO3 solution and once with water. After drying and concentration by evaporation, 197 mg of yellowish crystals remains.
Yield: 89%
Cld: C 41.85% H 4.21% N 13.01% O 18.58% S 22.35%
Fn~: C 40.44% H 3.93~ N 12.57% S 23.66%

Example 8 a) N-Benzyloxycarbonyl-0-t-butyloxycarbonylmethyl-~tyro~ine methylester) 3.29 g of the Z1-protected tyrosine methyl ester produced according to Example la) is dissolved in anhydrous DMF and mixed with 1.12 g of potassium-t-butylate. After 30 minutes, the solution of 1.95 g of bromoacetic acid-t-butyl ester is added in drops and heated for 4 hours to 110~C, then it is stirred for 4 more hours at room temperature. The mixture is added to water, extracted with CH2Cl2, washed, dried and concentrated by evaporation. After column chromatography (silica gel, CHzCl2/EE
19:1), 2.5 g of yellow oil remains.
Yield: 57%
Cld: C 65.00% H 6.59% N 3.16% 0 25.25%
Fnd: C 64.65% H 6.82% N 3.09%

b) 0-(t-Butyloxycarbonylmethyl)-tyro~ine methyl ester 1.5 g (3.4 mmol) of the Z1-protected compound produced according to Example 8a) is hydrogenated with hydrogen in 50 ml of ethyl acetate in the presence of 1.5 g of palladium on activated carbon (10%) at 50~C. After the reaction is completed, the catalyst is filtered out, and the solvent is drawn off. 1.0 g of colorless oil remains.
Yield: 99%
Cld: C 62.12% H 7.49% N 4.53% 0 25.86%
Fnd: C 61.88% H 7.67% N 4.47%

c) N-Toluenesulfonyl-O-t-butyloxycarbonylmethyl-tyrosine methyl ester The solution of 570 mg of toluenesulfonyl chloride in CH2Clz is added in drops to a solution of 920 mg of O-(t-butyloxycarbonylmethyl)-tyrosine methyl ester in CH2Cl2 at 0~C, and it is then slowly mixed with 300 mg of triethylamine and stirred overnight. It is poured onto ice water, extracted with CH2Cl2, the organic phase is washed twice with 10% HCl, twice with 10% NaHCO3 and twice with saturated common salt solution, dried and concentrated by evaporation. After treatment with ether, 900 mg of white crystals is obtained.
Yield: 65%
Cld: C 59.60% H 6.31% N 3.02% O 24.16% S 6.92%
Fnd: C 59.38% H 6.55% N 3.08% S 6.66%

d) N-Toluenesulfonyl-O-t-butyloxycarbonylmethyl-N-(2-aminoethyl)tyrosinamide The solution of 3.0 g of N-toluenesulfonyl-O-t-butyloxycarbonylmethyl-tyrosine methyl ester (6.5 mmol) in CH2Clz is added in drops to a solution of 5 g of ethylenediamine in 50 ml of CHzCl2. Then, it is refluxed for 4 hours. After cooling, it is mixed with water, the organic phase is separated and extracted several times with CH2Clz. The combined organic extracts are washed, dried and concentrated by evaporation.
After column chromatography (silica gel NeOH), l.S g of colorless oil remains.
Yield: 47%

Cld: C 58.64% H 6.77% N 8.55% O 19.53% S 6.52%
Fnd: C 58.39% H 6.91% N 8.38% S 6.56%

e) N-Toluenesulfonyl-O-carboxymethylt-N-~2-aminoethyl)tyrosinamide]
2 ml of trifluoroacetic acid is added at room temperature to a solution of 491 mg of the tert-butyl ester (1 mmol) produced according to Example 8d) in 25 ml of dichloromethane, and it is stirred at room temperature. After the reaction is completed, 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: 83%
Cld: C 55.16% H 5.79% N 9.65% O 22.04% S 7.36%
Fnd: C 53.88% H 5.64% N 10.02% S 7.44%

f) N-Toluenesulfonyl-O-carboxymethyl-tyrosine-[N-~2-~piperonylmercapto)-acetylaminoethyl)]amide 350 mg of piperonylmercaptoacetic acid-N-hydroxysuccinimidoester (1.08 mmol) in THF is slowly added in drops under argon and with exclusion of moisture to a solution of 435 mg of amine (1 mmol) produced according to Example 8e) in anhydrous THF at 0~C, and it is stirred for 2 hours at 0~C, then for 12 hours at room temperature. The solvent is drawn off, and the residue is chromatographed (silica gel, CH2Cl2/MeOH/HOAC

9:1:0.1) .
Yield: 58%

Cld: C 55.98% H 5.17% N 6.53% O 22.37% S 9.96%
Fnd: C S5.38% H 5.04% N 6.68% S 9.09%

g) N-Toluenesulfonyl-O-carboxymethyl-tyrosine-{N-t2-~mercaptoacetyl)-aminoethyl]}amide 644 mg of the protected S-compound (1 mmol) produced according to Example 8f) and a trace of anisole are added to 10 ml of trifluoroacetic acid at room temperature and heated briefly to boiling. After the reaction is completed, the trifluoroacetic acid is drawn off in a vacuum, the residue is taken up in a suitable solvent, washed with water, dried and concentrated by evaporation and chromatographed.
Yield: 34%
Cld: C 51.85% H 5.34% N 8.25% O 21.98% S 12.59 Fnd: C 51.62% H 5.53% N 8.65% S 13.61%

h) Tc-99m complex of N-toluenesulfonyl-O-carboxymethyl-tyrosine-{N-t2-(mercaptoacetyl)aminoethyl]}amide 1 mg of the compound produced according to Example 8g) is dissolved in 100 ~l of EtOH. 50 ~l of this solution is added to 250 ~l of a 0.1 M phosphate buffer of pH 8.5 and mixed with 50 ~l of a citrate solution (50 mg of trisodium citrate in 1 ml of water) and 2.5 ~l of a tin(II) chloride solution (5.0 mg of tin(II) chloride in 1 ml of 0.lN HCl). Then, it is mixed with 100 ~l of a 99m-Tc generator eluate and allowed to stand for 15 minutes. The labeling yield is determined with the aid of HPLC.

44 21 ~4561 Example 9 a) Cholesteryl diethylene glycol ~DEG-cholesterol) A solution of 54.1 g of cholesteryl toluenesulfonate (100 mmol) and 106 g of diethylene glycol in 250 ml of anhydrous dioxane is refluxed under nitrogen atmosphere (TLC control).
After the reaction is completed, it is mixed with water and extracted with CH2Cl2. After column chromatography (silica gel CH2Cl2/MeOH 9:1), a colorless solid remains.
Yield: 81%
Cld: C 78.43% H 11.46% 0 10.11%
Fnd: C 76.99% H 12.32%

b) Cholesteryl-2-chloroethyl(ethylene glycol) A solution of 4.75 g of DEG-cholesterol (10 mmol) produced according to Example 9a) in 50 ml of anhydrous carbon tetrachloride is mixed under nitrogen atmosphere with 3.14 g of pulverized triphenylphosphine (12 mmol) and refluxed. After cooling, it is diluted with 50 ml of petroleum ether or hexane and stored for some time at -20~C. The precipitate is suctioned off, and the procedure is again as above until no more precipitate settles. Then, it is dried and concentrated by evaporation. After column chromatography (silica gel CH2Cl2/MeOH
9:1), 3.70 g of an oil remains.
Yield: 75~
Cld: C 75.49% H 10.83% 0 6.49%
Fnd: C 74.73% H 11.74%

c) N-Benzyloxycarbonyl-2-t(4-cholesteryldiethylene-glycolyl)benzyl]-2-aminoacetic acid methyl ester The solution of 493 mg of the cholesterol derivative (1 mmol) (produced according to Example 9b) in 10 ml of toluene is added to a boiling solution of 295 mg of the N-benzyloxycarbonyl-tyrosine methyl ester (l mmol) (produced according to Example la) and 183 mg of potassium carbonate (1 mmol) in 10 ml of toluene, and it is refluxed for 6 hours. After the reaction is completed, it is allowed to cool to room temperature, concentrated by evaporation and chromatographed (silica gel petroleum ether/ethyl acetate 1:1) Yield: 29%
Cld: C 73.46% H 9.78% N 1.86% 0 14.89%
Fnd: C 73.28% H 10.01% N 1.70%

d) 2-[(4-Cholesteryldiethyleneglycolyl)benzyl~-2-aminoacetic acid methyl ester hydrochloride 7.52 g (10 mmol) of the Z-protected compound produced according to Example 9c) is dissolved in 50 ml of 3 M HCl in ethyl acetate and stirred for 6 hours at room temperature. After the solvent is drawn off, 6.02 g of white crystals remains.
Yield: 93%
Cld: C 71.53% H 9.66% N 2.04% 0 11.62%
Fnd: C 71.82% H 9.39% N 2.01%

e) N-p-Toluenesulfonyl-2[~4-cholesteryldiethyleneglycolyl)benzyl]-2-aminoacetic acid methyl ester 19.53 g of the amine (30 mmol) produced according to Example 9d) is dissolved in 50 ml of dichloromethane and mixed at 0~C
with 5.72 g of toluenesulfonic acid chloride in 30 ml of dichloromethane. While being stirred intensively, 3.0 g of triethylamine is added in drops at 0~C, and it is stirred for 1 hour at room temperature. After the reaction is completed, it is mixed with ice water and extracted several times with dichloromethane. The combined organic extracts are washed twice with cold 10% HCl, three times with 10% NaHC03 solution and twice with saturated NaCl solution. After drying, the solvent is drawn off and chromatographed (silica gel, CH2Cl2).
Yield: 77%
Cld: C 71.52% H 8.88% N 1.74% 0 13.89% S 3.98%
Fnd: C 70.89% H 9.02% N 1.66% S 3.70%

f) N-p-Toluenesulfonyl-2[~4-cholesteryldiethyleneglycolyl)benzyl]-2-aminoacetic acidtN-(2-aminoethyl)]amide A solution of 806 mg of the tosylglycine ester (1 mmol) produced according to Example 9e in 1 ml of anhydrous dichloromethane is slowly added in drops to a solution of 1.2 g of ethylenediamine (20 mmol) in 1 ml of anhydrous dichloromethane, and it is refluxed. After the reaction is , completed, it is concentrated by evaporation in a vacuum, and the residue is chromatographed (silica gel, MeOH).
Yield: 85%
Cld: C 70.55% H 9.06% N 5.04% O 11.51% S 3.84%
Fnd: C 69.24% H 9.31~ N 4.83% S 3.71%

g) N-p-Toluenesulfonyl-2tt4-cholesteryldiethyleneglycolyl)-benzyl]-2-aminoacetic acid-N-{2-Nt(3-carboxy-2-mercaptoacetyl-l-oxopropyl)]aminoethyl}amide 0.38 g (2.2 mmol) of 2-acetylmercaptosuccinic anhydride is added to a solution of 1.83 g of the amine (2.2 mmol) produced according to Example 9f) in 10 ml of pyridine/DMF (50:50), and it is stirred for 4 hours at room temperature. Then, the solvent is drawn off, the residue is taken up in 0.5N HCl and extracted with CH2Cl2. After flash chromatography, 887 mg of a yellow oil remains.
Yield: 40%
Cld: C 65.51% H 8.10% N 4.17% O 15.87% S 6.36%
Fnd: C 63.66% H 7.58% N 3.99% S 7.41%

h) Tc-99m complex of N-p-toluenesulfonyl-2t~4-cholesteryldiethyleneglycolyl)-benzyl]-2-aminoacetic acid-N-{2-Nl(3-carboxy-2-mercaptoacetyl-1-oxopropyl)]-aminoethyl}amide 1 mg of the ligand produced according to Example 9g is dissolved in 100 ~l of ethanol. 50 ~l of this solution is added to 250 ~l of a 0.1 M phosphate buffer (pH 8.5) and then mixed with 50 ~l of a citrate solution (50 mg of trisodium citrate in 1.0 ml of water) and 2.5 ~l of a tin(II) chloride solution (5.0 mg of tin(II) chloride in 1 ml of O.lN HCl). Then, it is mixed with 50 ~l of a Tc-99m generator eluate and allowed to stand for 15 minutes. The labeling yield is determined with the aid of HPLC.

Example 10 a) l-Tosyl-1,4,7-triazaheptan-3-one A solution of 25.73 g of tosylglycine methyl ester (0.1 mol) in anhydrous dichloromethane is slowly added in drops to a solution of 30 g of ethylenediamine (0.5 mol) in 50 ml of anhydrous dichloromethane. It is stirred for 12 hours at room temperature and then concentrated by evaporation in a vacuum. 24 g of a yellow oil remains.
Yield: 88%
Cld: C 48.69% H 6.32% N 15.49% 0 17.69% S 11.82%
Fnd: C 47.23% H 6.67% N 15.34% S 11.54%

b) 7-N-tert-Butoxycarbonyl-1-tosyl-1,4,7-triazaheptan-3-one A solution of 27 g (100 mmol) of the amino compound produced according to Example lOa) in 500 ml of dioxane is mixed in one portion with 74 g of di-tert-butyl-dicarbonate (340 mmol). The solution is stirred for 3 hours at room temperature, ~hen poured onto ice water and extracted three times with ethyl acetate. The combined organic phases are washed three times with water and once with saturated common salt solution, dried (MgSO4), concentrated by evaporation and recrystallized. 27.9 g of white crystals remains.
Yield: 75%
Cld: C 51.74~ H 6.78% N 11.31% 0 21.54% S 8.63%
Fnd: C 51.41% H 6.96% N 11.12% S 8.91%

c) 7-N-t-Butyloxycarbonyl-1-n-hexyl-1-tosyl-1,4,7-triazaheptan-3-one 5 g of potassium carbonate is added to a solution of 3.71 g of the sulfonamide (10 mmol) produced according to Example lOb) in 50 ml of DMF at room temperature, and then the solution of 2.54 g of hexyl iodide (12 mmol) in 10 ml of DMF is added in drops and heated for 3 hours to 110~C. After the reaction is completed, it is allowed to cool to room temperature, poured onto ice, extracted with CH2Cl2, washed several times with water, dried, filtered and concentrated by evaporation. The residue is recrystallized from ethyl acetate.
Yield: 89%
Cld: C 58.00% H 8.19% N 9.22% 0 17.56% S 7.04%
Fnd: C 57.37% H 7.87% N 9.01% S 6.95%

d) l-n-Hexyl-1-tosyl-1,4,7-triazaheptan-3-one 456 mg of the sulfonamide (1 mmol) produced according to Example lOc) is added to 10 ml of trifluoroacetic acid at 0~C, and then it is stirred for 2 hours at room temperature. After the reaction is completed, it is poured onto ice, weakly alkalized (NaHCO3), extracted, concentrated by evaporation and dried.
Yield: 94%
Cld: C 57.44% H 8.22% N 11.82% O 13.50% S 9.02%
Fnd: C 57.96% H 8.01% N 11.63% S 8.75%

e) 10-Acetyl-9-carboxymethyl-1-n-hexyl-1-to~yl-10-thia-1,4,7-triazadecane-3,8-dione The solution of 3.55 g (lO mmol) of the amino compound in 10 ml of DMF is slowly added in drops to a solution of 1.91 g of acetylmercaptosuccinic anhydride (20 mmol) in 20 ml of DMF at room temperature, and it is stirred overnight. Then, it is poured onto semiconcentrated HCl, extracted with CH2Cl2 and chromatographed (silica gel, EtOAc/MeOH from 9:1 to 1:1).
Yield: 66%
Cld: C 52.16% H 6.66% N 7.93% O 21.15% S 12.11%
Fnd: C 52.85% H 6.87% N 7.52% S 11.66%

f) Tc-99m complex of 10-Acetyl-9-carboxymethyl-1-n-hexyl-1-tosyl-10-thia-1,4,7-triazadecane-3,8-dione 1 mg of N,N thexyl-p-toluenesulfonyl]glycyl-N-[(3-carboxy-2-mercaptoacetyl-1-oxopropyl)]aminoethyl}amide is dissolved in 100 ~l of EtOH. 50 ~l of this solution is diluted with 100 ~l of EtOH and mixed with 100 ~l of a 0.1 M phosphate buffer at pH 7.5 and 100 ~l of a Tc-99m-gluconate solution. The labeling yield is > 95% (silica gel, 95% EtOH) 2 1 ~4561 Example 11 a) N-~-Toluenesulfonyl-N-~-tert-butyloxycarbonyllysine methyl ester 1.91 mg of toluenesulfonyl chloride is added in portions to a solution of 2.60 g of N-~-tert-butyloxycarbonyllysine methyl ester (10 mmol) in 50 ml of anhydrous pyridine at 0~C, and it is allowed to stand for 24 hours at 4~C. Then, it is poured onto ice water, and the precipitate is separated and recrystallized.
Yield: 78%
Cld: C 55.05% H 7.30% N 6.76% 0 23.16% S 7.74%
Fnd: C 54.67% H 7.42% N 6.64% S 7.54%

b) N-~-Toluenesulfonyl-N-~-tert-butyloxycarbonyllysine-[N-~2-aminoethyl)]amide A solution of 415 mg of the tosyllysine ester (1 mmol) produced according to Example lla) in 1 ml of anhydrous dichloromethane is slowly added in drops to a solution of 600 mg of ethylenediamine (10 mmol) in 1 ml of anhydrous dichloromethane and refluxed. After the reaction is completed, it is concentrated by evaporation in a vacuum, and the residue is recrystallized.
Yield: 59%
Cld: C 54.28% H 7.74% N 12.66% 0 18.08% S 7.25%
Fnd: C 54.54% H 7.57% N 12.18% S 7.06%

c) N-~-Toluenesulfonyl-N-~-tert-butyloxycarbonyllysine-[N-(2-~piperonylmercapto)-acetylaminoethyl)]amide 350 mg of piperonylmercaptoacetic acid-N-hydroxysuccinimidoester (1.08 mmol) in THF is slowly added in drops under argon and with exclusion of moisture-to a solution of 443 mg of amine (1 mmol) produced according to Example llb) in anhydrous THF at 0~C, and it is stirred for 2 hours at 0~C, then for 12 hours at room temperature. The solvent is drawn off, and the residue is chromatographed (silica gel, CH2Cl2/MeOH 9:1).
Yield: 54%
Cld: C 55.37% H 6.51% N 8.61% 0 19.67% S 9.85%
Fnd: C 55.22% H 6.85% N 8.41% S 9.66%

d) N-~-Toluenesulfonyl-lysine-[N-(2-(piperonylmercapto)acetylaminoethyl)]amide 651 mg of N-~-toluenesulfonyl-N-~-tert-butyloxycarbonyllysine-[N-(2-(piperonylmercapto)-acetylaminoethyl)]amide is dissolved in 5 ml of 3 M HCl in ethyl acetate, and it is stirred for 6 hours at room temperature.
After the solvent is drawn off, 530 mg of white crystals remains.
Yield: 96%
Cld: C 54.63% H 6.05~ N 10.19% 0 17.46% S 11.67 Fnd: C 54.40% H 6.47~ N 10.01% S 11.84%

Example 12 a) N-~-Toluenesulfonyl-N-~-tert-butyloxycarbonyllysine-tN-~2-benzoylmercapto)-acetylaminoethyl)]amide 316 mg of benzoylmercaptoacetic acid-N-hydroxysuccinimidoester (1.08 mmol) in tetrahydrofuran is slowly added in drops under argon and with exclusion of moisture to a solution of 443 mg of amine produced according to Example llb) in anhydrous tetrahydrofuran at 0~C, and it is stirred for 2 hours at 0~C, then for 12 hours at room temperature. The solvent is drawn off, and the residue is chromatographed (silica gel, CH2Clz/MeOH 9:1).
Yield: 65%
Cld: C 56.11% H 6.50% N 9.03% 0 18.04% S 10.33%
Fnd: C 55.98% H 6.86% N 8.88% S 10.05%

b) N-~-Toluenesulfonyl-lysine-tN-(2-benzoylmercapto)acetylaminoethyl)lamide 621 mg of the title compound of Example 12a) is dissolved in 5 ml of 3 M HCl in ethyl acetate, and it is stirred for 6 hours at room temperature. After the solvent is drawn off, 500 mg of white crystals remains.
Yield: 90%
Cld: C 51.74% H 5.97% N 10.06% 0 14.36% S 11.51%
Fnd: C 51.40% H 6.14% N 10.10% S 11.48%

54 21 q4561 c) Tc-99m complex of N-~-toluenesulfonyl-lysine-lN-~2-~benzoylmercapto)-acetylaminoethyl)]amide 10 mg of the amide produced according to Example 12b) is mixed with 500 ~1 of lN NaOH and allowed to stand for lS minutes.
50 ~1 of this solution is added to 250 ~l of a 0.1 M phosphate buffer of pH 8.s and mixed with 50 ~l of a citrate solution (So mg of trisodium citrate in 1.0 ml of water) and 2.5 ~1 of a tin(II) chloride solution (5.0 mg of tin(II) chloride in 1.0 ml of 0.lN HCl). Then, it is mixed with 50 ~1 of a Tc-99m generator eluate and allowed to stand for 15 minutes. The labeling yield is determined with the aid of HPLC (> 95%).

Example 13 a) N-t3-~N-tert-butyloxycarbonyl)aminobenzenesulfonyl]-glycine methyl ester A solution of 2.12 g (10 mmol) of the amino compound produced according to Example 7b) and 3 g of triethylamine (30 mmol) in 50 ml of dioxane are mixed in one portion with 7.4 g of di-tert-butyldicarbonate (34 mmol). (Gas generation!) The solution is stirred for 30 minutes at room temperature, then poured onto ice water and extracted three times with ethyl acetate. The combined organic phases are washed three times with water and once with saturated common salt solution, dried (MgSO4), concentrated by evaporation and recrystallized.
Yield: 85%
Cld: C 48.83% H s.85% N 8.13% o 27.88% S 9.31%
Fnd: C 47.44% H 5.93% N 8 . 57% S 9 . 66%

b) N-t3-(N-tert-butyloxycarbonyl)aminobenzenesulfonyl]-glycyl-[N'-(2-aminoethyl)amide A solution of 12.11 g of the glycine ester produced according to Example 13a) (35 mmol) in 250 ml of anhydrous toluene/dioxane or optionally only dioxane is slowly added in drops to a boiling solution of 42 g of ethylenediamine (700 mmol) in 100 ml of anhydrous toluene, and it is refluxed. After the reaction is completed, it is concentrated by evaporation in a vacuum, and the residue is taken up in chloroform and washed with water.
Yield: 78%
Cld: C 44.06% H 6.16% N 13.70% O 19.56% S 7.84%
Fnd: C 45.44% H 6.63% N 14.57% S 7.66%

c) N-~3-(N-tert-Butyloxycarbonyl)aminobenzenesulfonyl]-glycyl-~N'-tt8-benzoylmercapto)acetyl]-(2-aminoethyl)}amide 5 mmol of the S-benzoylmercaptoacetic acid (0.98 g) and 10 mmol of NEt3 (1.4 ml) and 5 mmol of the amine produced according to Example 13b) (1.86 g) are mixed with 50 ml of dichloromethane and cooled to 10~C. Then, 5.5 mmol (1.375 g) of BOP-Cl is added and stirred while being cooled in water. (After 10-20 minutes, a clear solution is obtained.) Then, it is stirred, stirred for 1 more hour, mixed with water and brought to pH 1-1.5 with 4N HCl.
Yield: 62%
Cld: C 52.16% H 5.84% N 10.14% O 20.27% S 11.60%
Fnd: C 53.44% H 6.63% N 10.57% S 11.66%

d) N-[3-Aminobenzenesulfonyl]-glycyl-{N'-t(S-benzoylmercapto)acetyl]-~2-aminoethyl)}amide A freshly prepared solution of 3 M HCl in EtOAc (10 ml, 30 mmol) is added to a solution of 552 mg of the protected amine produced according to Example 13c) (1 mmol) in 5 ml of EtOAc. It is stirred for 30 minutes at room temperature. The solvent is drawn off, and the residue is dried in a vacuum at room temperature.
Yield: 93%
Cld: C 46.86% H 4.76% N 11.51% O 16.43% S 13.17%
Fnd: C 45.44~ H 5.33% N 10.57% S 13.66%

e) N-[3-Isothiocyanatobenzene~ulfonyl]-glycyl-{N'-t(S-benzoylmercapto)acetyl]-(2-aminoethyl)}amide The solution of 65 mg of thiophosgene in a little dichloromethane is added in drops with exclusion of moisture to a solution of 200 mg of N-[(3-aminosulfonyl)]glycyl-{N'-[(acetylmercapto)-acetyl](2-aminoethyl)}amide produced according to Example 13d) and 20 ~l of triethylamine in 4 ml of dichloromethane, and it is stirred for 4 hours at room temperature. After the solvent is drawn off, a residue remains, which is taken up in chloroform and washed with 0.1% citric acid, washed twice with sodium chloride solution and once with water, dried, concentrated by evaporation and chromatographed.
Yield: 66%

Cld: C 48.77% H 4.09% N 11.37% O 16.24% S 19.53%
Fnd: C 48.44% H 4.63% N 11.75% S 20.06%

2 I q456 1 Example 14 a) N-Toluenesulfonyl-O-methoxycarbonylmethyl[-N-(2-~minoethyl)tyro~inamide]
HCl gas is introduced until saturation in a solution of4.35 g of amine produced according to Example 8e) (10 mmol) in 100 ml of anhydrous methanol, and it is stirred overnight. After the solvent is removed, a crystalline residue remains.
Yield: 92%
Cld: C 51.90% H 5.81% N 8.65% 0 19.75% S 6.60%
Fnd: C S1.59% H 5.93% N 8.47% S 6.49%

b) N-Toluenesulfonyl-0-methoxycarbonylmethyltyro~ine-[N-(2-tritylmercapto)-acetylaminoethyl)]amide 4.66 g of S-tritylmercaptoacetic acid-N-hydroxysuccinimidoester (10.8 mmol) in tetrahydrofuran is slowly added in drops under argon to a solution of 4.35 g of amine produced according to Example 14a) (10 mmol) and 2.02 g of triethylamine in tetrahydrofuran/water at 0~C, and it is stirred for 2 hours at 0~C, then for 12 hours at room temperature. The -solvent is drawn off, and the residue is chromatographed (silica gel, CH2C12)-Yield: 68%
Cld: C 65.86% H 5.66% N 5.49% 0 14.62% S 8.37%
Fnd: C 65.66% H 5.71% N 5.36% S 8.21%

c) N-Toluenesulfonyl-O-carboxymethyltyrosine-tN-(2-tritylmercapto)acetylaminoethyl)]amide The solution of 766 mg of the methyl ester produced according to Example 14b) (1 mmol) in 2.5 ml of methanol is added to a methanolic KOH solution and stirred at room temperature.
After the reaction is completed, the solvent is drawn off, the potassium salt is taken up in water, weakly acidified and extracted with chloroform. The organic extract is washed with water, dried and concentrated by evaporation.
Yield: 63%
Cld: C 65.49% H 5.50% N 5.59% O 1490~ S 8.53%
Fnd: C 65.26% H 5.72% N 5.43% S 8.43%

d) N-Toluenesulfonyl-~(HOOC-Trp-Ile-Ile Asp-Leu-His Gly-NH)-carbonylmethyltyrosine-tN-(2-tritylmercapto)acetylaminoethyl)]amide 206 mg of dicyclohexylcarbodiimide (1 mmol) dissolved in 2 ml of dimethylformamide is added in drops to a solution of 1 mmol of the acid (980 mg) produced according to Example 14c) and 115 mg of N-hydroxysuccinimide (1 mmol) in 2 ml of anhydrous dimethylformamide at 0~C within 5 minutes. It is stirred first for another 30 minutes at 0~C, then the solution of 1 mmol of the peptide (853 mg) H2N-Gly-His-Leu-Asp-Ile-Ile-Trp (produced analogously to Barany and Merrifield, The Peptides: Analysis, Biology, Academic Press New York, 1980; Steward and Young, Solid Phase Peptides Syntheses, 2nd ed.; Pierce Chemical W., Rockford, II, 1984) and 304 mg (3 mmol) of triethylamine in 10 ml of anhydrous dimethylformamide is added under argon atmosphere within 30 minutes. It is stirred for 12 more hours at room temperature, 200 ~l of glacial acetic acid is added, stirred again for 30 minutes, the product is filtered off from N,N'-dicyclohexylurea, and the residue is concentrated by evaporation.
After stirring up with diethyl ether, a white residue remains, which is recrystallized from dimethylformamide/ether.
Yield: 35%
Cld: C 62.07% H 6.29% N 4.04% O 16.13% S 4.04%
Fnd: C 61.77% H 6.52% N 4.32% S 4.34%

e) N-Toluenesulfonyl-[(HOOC-Trp-Ile-Ile Asp-Leu-Hi~ Gly-NH)carbonyl]-methyltyroqine-[N-(mercapto)acetylaminoethyl)]amide 530 mg of the protected S-compound produced according to Example 14d) (0.3 mmol) and a trace of anisole are added to 10 ml of trifluoroacetic acid under argon atmosphere at room temperature and heated briefly to boiling. Then, the trifluoroacetic acid is drawn off in a vacuum, the residue is taken up in tetrahydrofuran, concentrated by evaporation and chromatographed.
Yield: 59%
Cld: C 56.67% H 6.32~ N 13.42% O 18.87% S 4.73%
Fnd: c 56.14~ H 6.48~ N 12.98~ S 4.81%

f) Tc-99m complex of N-toluenesulfonyl-[~HOOC-Trp-Ile-Ile Asp-~eu-Hi-Q Gly-NH)carbonylmethyltyro-Qine-tN-(mercaptoacetyl)aminoethyl)]amide 1 mg of the above-named compound (Example 14e) is dissolved in 100 ~l of EtOH/water 1:1. 50 ~l of this solution is mixed with 100 ~l of a 0.1 M phosphate buffer of pH 9.5 and 100 ~1 of a Tc-99m-gluconate solution. The labeling yield is > 95% (HPLC, LiChrospher RP18, H2O/MeCN + 0.1% TFA).

Example 15 a) N-Toluenesulfonyl-O-t-butyloxycarbonylmethyltyrosine-tN-(2-benzoylmercapto)-acetylaminoethyl)]amide 2 mmol of S-benzoyl-2-mercaptoacetic acid (394 mg) and 4 mmol of NEt3 (560 ~1) and 2 mmol (982 mg) of the compound obtained according to Example 8d) are mixed with 5 ml of dichloromethane and cooled to 10~C. Then, 2 mmol (509 mg) of BOP-Cl is added and stirred for 12 hours while being cooled in water, then mixed with water and brought to pH 1-1.5 with 4N HCl.
Then, it is extracted with dichloromethane, the combined organic extracts are washed with NaHCO3 and water, dried, concentrated by evaporation and chromatographed.
Yield: 76%
Cld: C 59.18% H 5.87% N 6.27% O 19.11% S 9.58%
Fnd: c 60.88~ H 5.89% N 5.63% S 8.74%

b) N-Toluenesulfonyl-O-carboxymethyltyrosine-tN-(2-benzoylmercapto)acetyl-aminoethyl)]amide 644 mg of the protected S-compound produced according to Example 15a) (1 mmol) is added to 10 ml of trifluoroacetic acid at room temperature, and it is stirred at room temperature.
After the reaction is completed, the trifluoroacetic acid is drawn off in a vacuum, the residue is taken up in ethyl acetate, washed with water, dried and concentrated by evaporation and chromatographed.
Yield: 77%
Cld: C 56.76% H 5.09% N 6.85% 0 20.86% S 10.45%
Fnd: C 57.14% H 5.68% N 7.23% S 11.31~

c) N-Toluenesulfonyl-[(HOOC-Trp-Ile-Ile Asp-~eu-(D-Trp)-NH)carbonylmethyl-tyrosine-[N-(2-benzoylmercapto)acetylaminoethyl)1amide 206 mg of dicyclohexylcarbodiimide (1 mmol) dissolved in 2 ml of tetrahydrofuran is added in drops to a solution of 1 mmol of acid (614 mg) produced according to Example 15b) and 115 mg of N-hydroxysuccinimide (1 mmol) in 2 ml of anhydrous tetrahydrofuran at 0~C within 5 minutes. First, it is stirred for another 30 minutes at 0~C, then the solution of 1 mmol of the peptide (845 mg) of H2N-(D-Trp)-Leu-Asp-Ile-Ile-Trp (produced analogously to Barany and Merrifield, The Peptides: Analysis, Biology, Academic Press, New York, 1980; Steward and ~oung, Solid Phase Peptides Syntheses, 2nd ed.; Pierce Chemical W., Rockford, Il 1984) and 304 mg (3 mmol) of triethylamine in 10 ml of anhydrous dimethylformamide are added under argon atmosphere within 30 minutes. It is stirred for 12 more hours at room temperature, 200 ~l of glacial acetic acid is added, stirred again for 30 minutes, the product is filtered off from N,N'-dicyclohexylurea, and the residue is extracted twice with boiling tetrahydrofuran. The combined filtrates are evaporated to dryness and chromatographed (silica gel, CH2Cl2).
Yield: 40%
Cld: C 60.86% H 6.23% N 10.69% O 17.77% S 4.45%
Fnd: C 60.77% H 6.42% N 10.32% S 4.65%

d) Tc-99m complex of N-toluene~ulfonyl-t(HOOC-Trp-Ile-Ile Asp-Leu-(D-Trp)-NH)carbonylmethyltyro~ine-tN-(2-mercaptoacetyl)aminoethyl)]amide 2 mg of the compound produced according to Example 15c) is dissolved in 100 ~l of EtOH and mixed with 100 ~l of lN NaOH.
After 15 minutes, 50 ~l of this solution is added to 250 ~l of a 0.1 M phosphate buffer of pH 8.5 and mixed with 50 ~l of a citrate solution (50 mg of trisodium citrate in 1.0 ml of water) and 2.5 ~l of a tin(II) chloride solution (5.0 mg of tin(II) chloride in 1.0 ml of 0.lN HCl). Then, it is mixed with 50 ~l of a Tc-99m generator eluate and allowed to stand again for 15 minutes. The labeling yield (> 95%) is determined with the aid of HPLC.

...
Example 16 a) N-Tosyl-tyro~inemethyle~ter-4-benzyl ether A solution of 32.58 g (171 mmol) of p-toluenesulfonic acid chloride in 100 ml of pyridine is added in drops to 50 g (155.37 mmol) of tyrosinemethylester-4-benzyl ether hydrochloride in 300 ml of pyridine at 0~C, and it is stirred for 3 hours at 0~C. It is evaporated to dryness in a vacuum, and the residue is dissolved in 500 ml of methylene chloride. The organic phase is shaken out twice with 300 ml of 5N hydrochloric acid. The organic phase is dried on magnesium sulfate and concentrated by evaporation in a vacuum. The residue is recrystallized from a little methanol. 68.29 g of a colorless crystalline powder is obtained.
Yield: 93%
Cld: C 65.58% H 5.73% N 3.19% S 7.29%
Fnd: C 65.30% H 5.81% N 3.02% S 7.18%

b) 2-t4-Benzyloxybenzyl)-1-tosyl-1,4,7-triazaheptan-3-one 45 g (102.38 mmol) of the title substance of Example 16a) is introduced in 1 l of 1,2-diaminoethane within 1 hour and then stirred for 3 hours at 80~C. The residue is evaporated to dryness, and the residue is absorptively precipitated in 200 ml of water. The precipitate is suctioned off and rewashed with a great deal of water. Then, it is dried overnight in a vacuum at 60~C. 46.91 g of a cream-colored, amorphous powder is obtained.
Yield: 98%
Cld: C 64.22% H 6.25% N 8.95% S 6.86%

Fnd: C 64.05% H 6.17% N 9.05% S 6.78%

c) 9-Chloro-2-(~-benzyloxybenzyl)-1-tosyl-1,4,7-triaza-nonane-3,8-dione lO g (21.39 mmol) of the title compound of Example 16b) is dissolved in 100 ml of chloroform, and 2.38 g (23.53 mmol) of triethylamine is added. At 0~C, 2.66 g (23.53 mmol) of chloroacetyl chloride in 20 ml of chloroform is added in drops within 30 minutes. It is stirred for 30 minutes at 0~C. 200 ml of lN hydrochloric acid is added and shaken vigorously. The organic phase is separated, dried on magnesium sulfate and concentrated by evaporation in a vacuum. The residue is recrystallized from a little methanol. 10.36 g of a cream-colored, crystalline solid is obtained.
Yield: 89%
Cld: C 59.61% H 5.56% N 7.72% S 5.89% Cl 6.52%
Fnd: C 59.50% H 5.69% N 7.55% s 5.71% Cl 6.38%

d) 10-Benzoyl-2-~4-benzyloxybenzyl)-1-tosyl-10-thia-1,4,7-triazadecane-3,8-dione 9 g (16.54 mmol) of the title compound of Example 16c) is dissolved in 100 ml of chloroform, and 1.67 g (16.54 mmol) of triethylamine is added. Then, 2.29 g (16.54 mmol) of thiobenzoic acid is added and refluxed for 10 minutes. It is cooled to room temperature and shaken out once with 2N hydrochloric acid and once with a 5% sodium carbonate solution. The organic phase is dried on magnesium sulfate and concentrated by evaporation in a vacuum. The residue is recrystallized from a little methanol.
9.61 g of a colorless, crystalline powder is obtained.
Yield: 90%
Cld: C 63.24% H 5.46% N 6.51% S 9.93%
Fnd: C 63.15% H 5.57% N 6.40% S 9.81%

Example 17 a) 10-Acetyl-2-~4-benzyloxybenzyl)-1-to~yl-10-thia-1,4,7-triazadecane-3,8-dione 9 g (16.54 mmol) of the title compound of Example 16c) is dissolved in 100 ml of chloroform, and 1.67 g (16.54 mmol) of triethylamine is added. Then, 1.28 g (16.54 mmol) of thioacetic acid is added and refluxed for 10 minutes. It is cooled to room temperature, shaken out with 2N hydrochloric acid and then with a 5% sodium carbonate solution. The organic phase is dried on magnesium sulfate and concentrated by evaporation in a vacuum.
The residue is recrystallized from a little acetone. 8.20 g of cream-colored crystals is obtained.
Yield: 85%
Cld: C 59.67% H 5.70% N 7.20% S 10.98%
Fnd: C 59.51% H 5.81% N 7.05% S 10.80%

Example 18 a) lo-Trifluoroacetyl-2-~4-benzyloxybenzyl)-1-tosyl-lo-thia-1,4,7-triazadecane-3,8-dione 9 g (16.54 mmol) of the title compound of Example 16c) is dissolved in 100 ml of chloroform and 1.67 g (1654 mmol) of trifluoromethylthioacetic acid, and it is refluxed for 10 minutes. It is cooled to room temperature, shaken out with 2N
hydrochloric acid and then with a 1% sodium carbonate solution.
The organic phase is dried on magnesium sulfate and concentrated by evaporation in a vacuum. The residue is recrystallized from a little acetone/ether. 8.75 g of colorless crystals is obtained.
Yield: 83%
Cld: C 54.62% H 4.74% N 6.59% S 10.05% F 8.94%
Fnd: C 54.47% H 4.61% N 6.50% S 9.90% F 8.81%

Example 19 a) N-Mesyl-tyrosinemethylester-4-benzyl ether 19.58 g (171 mmol) of methanesulfonic acid chloride is added in drops to 50 g (155.37 mmol) of tyrosinemethylester-4-benzyl ether hydrochloride in 300 ml of pyridine at 0~C, and it is stirred for 3 hours at 0~C. It is evaporated to dryness in a vacuum, and the residue is dissolved in 500 ml of methylene chloride. The organic phase is shaken out twice with 300 ml of 5N hydrochloric acid. The organic phase is dried on magnesium sulfate and concentrated by evaporation in a vacuum. The residue is recrystallized from a little methanol. 53.64 g of a colorless, crystalline powder is obtained.
Yield: 95%
Cld: C 59.49% H 5.82% N 3.85% S 8.82%
Fnd: C 59.30% H 5.95% N 3.71% S 8.70%

b) 2-(4-Benzyloxybenzyl)-1-mesyl-1,4,7-triazaheptan-3-one 37.2 g (102.38 mmol) of the title substance of Example l9a) is introduced in 1 liter of 1,2-diaminoethane within 1 hour and then stirred for 3 hours at 80~C. The residue is evaporated to dryness, and the residue is absorptively precipitated in 200 ml of water. The precipitate is suctioned off and rewashed with a great deal of water. Then, it is dried overnight in a vacuum at 60~C. 37.68 g of a cream-colored, amorphous powder is obtained.
Yield: 97%
Cld: C 56.97% H 6.64% N 11.07% S 8.45%
Fnd: C 56.81% H 6.72% N 10.93% S 8.32%

c) 9-Chloro-2-(4-benzyloxybenzyl)-1-mesyl-1,4,7-triaza-nonane-3,8-dione 10 g (26.35 mmol) of the title compound of Example l9b) is dissolved in 100 ml of chloroform, and 2.93 g (28.99 mmol) of triethylamine is added. At 0~C, 3.27 g (28.99 mmol) of chloroacetyl chloride in 20 ml of chloroform is added in drops within 30 minutes. It is stirred for 30 minutes at 0~C. 200 ml of lN hydrochloric acid is added and vigorously shaken. The organic phase is separated, dried on magnesium sulfate and concentrated by evaporation in a vacuum. The residue is recrystallized from a little methanol. 10.93 g of a cream-colored, crystalline solid is obtained.
Yield: 91%
Cld: C 52.68% H 5.75% N 9.22% S 7.03% Cl 7.78%
Fnd: C 52.51% H 5.82% N 9.13% S 6.90% Cl 7.68%

....

d) 10-Benzoyl-2-(4-benzyloxybenzyl)-1-mesyl-10-thia-1,4,7-triaza-decane-3,8-dione 7.54 g (16.54 mmol) of the title compound of Example 16c) is dissolved in 100 ml of chloroform, and 1.67 g (16.S4 mmol) of triethylamine is added. Then, 2.29 g (16.54 mmol) of thiobenzoic acid is added and refluxed for 10 minutes. It is cooled to room temperature and shaken out once with 2N hydrochloric acid and once with a 5% sodium carbonate solution. The organic phase is dried on magnesium sulfate and concentrated by evaporation in a vacuum. The residue is recrystallized from a little methanol.

8.11 g of a colorless, crystalline powder is obtained.
Yield: 88%
Cld: C 58.15% H 5.60% N 7.53% S 11.50%
Fnd: C 58.03% H 5.71% N 7.61% S 11.38%

Example 20 a) 9-Chloro-2-~4-hydroxybenzyl)-1-tosyl-1,4,7-triazanonane-3,8-dione 20 g (36.76 mmol) of the title compound of Example 16c) is dissolved in 200 ml of methylene chloride, and 2 ml of glacial acetic acid is added. Then, 3 g of palladium catalyst (10% Pd/C) is added and hydrogenated overnight. The catalyst is filtered out, and it is evaporated to dryness in a vacuum. 16.52 g of an amorphous solid is obtained.
Yield: 99%
Cld: C 52.92% H 5.33% N 9.26% S 7.06% Cl 7.81%
Fnd: C 52.81% H 5.26% N 9.11% S 6.93% Cl 7.72%

b) 9-Chloro-2-(4-palmitoyloxybenzyl)-l-tosyl-1,4,7-triazanonane-3,8-dione 10 g (22.03 mmol) of the title compound of Example 20a) is dissolved in 100 ml of chloroform, and 2.45 g (23.53 mmol) of triethylamine is added. At 0~C, 6.66 g (24.23 mmol) of palmitic acid chloride is added in drops within 10 minutes, and it is stirred for 2 more hours at this temperature. It is shaken out with 200 ml of 2N hydrochloric acid, the organic phase is dried on magnesium sulfate and concentrated by evaporation. The residue is chromatographed on silica gel (mobile solvent:
methylene chloride/acetone = 20:1). 11.90 g of a waxy solid is obtained.
Yield: 78%
Cld: C 62.45% H 7.86% N 6.07% S 4.63% Cl 5.12%
Fnd: C 62.28% H 7.70% N 5.89% S 4.54% Cl 4.98%

c) lO-Benzoyl-2-~4-palmitoyloxybenzyl)-l-tosyl-lO-thia-1,4,7-triazadecane-3,8-dione 8 g (11.55 mmol) of the title compound of Example 20b) is dissolved in 100 ml of chloroform and 1.17 g (11.55 mmol) of triethylamine is added. Then, 1.60 g (11.55 mmol) of thiobenzoic acid is added and refluxed for 10 minutes. It is cooled to room temperature and shaken out once with 2N hydrochloric acid and once with a 5% sodium carbonate solution. After drying on magnesium sulfate, the organic phase is concentrated by evaporation in a vacuum, and the residue is chromatographed on silica gel (mobile solvent: methylene chloride/hexane/

acetone:20:10:1). 8.53 g of colorless flakes (of ether) is obtained.
Yield: 93%
Cld: C 65.04% H 7.49% N 5.29% S 8.07%
Fnd: C 64.90% H 7.55% N 5.17% S 7.91%

Example 21 a) Phenolester of 3-cholesterol ~uccinic acid semi-e~ter with 9-chloro-2-~4-hyd~oxybenzyl)-1-tosyl-1,4,7-triazanonane 3,8-dione 10 g (22.03 mmol) of the title compound of Example 20a) and 10.72 g (22.03 mmol) of cholesterol succinic acid semi-ester are dissolved in 49 ml of dimethylformamide and cooled to 0~C. Then, 300 mg of 4-dimethylaminopyridine and 5.45 g (26.44 mmol) of dicyclohexylcarbonyl are added and stirred for 3 hours at 0~C.
It is allowed to stir at room temperature overnight. 50 ml of ether is added and precipitated urea is filtered out, the filtrate is diluted with 250 ml of ethyl acetate and shaken out 5 times with 200 ml of water. The organic phase is dried on magnesium sulfate and concentrated by evaporation in a vacuum.
The residue is chromatographed on silica gel (mobile solvent:
methylene chloride/hexane/ethyl acetate = 10:5:1). 17.60 g of a waxy solid is obtained.
Yield: 78~
Cld: C 66.39% H 7.87% N 4.55% S 3.48% Cl 3.84%
Fnd: C 66.28% H 7.95% N 4.47% S 3.31% Cl 3.70%

71 21 ~4561 b) Phenolester of cholesterol succinic acid semi-ester with 10-benzoyl-2-(4-hydroxybenzyl)1-tosyl-10-thia-1,4,7-triazadecane-3,8-dione 6 g (6.50 mmol) of the title compound of Example 21a) is dissolved in 50 ml of chloroform and 0.66 g (6.50 mmol) of triethylamine is added. Then, 0.9 g (6.50 mmol) of thiobenzoic acid is added and refluxed for 10 minutes. It is cooled to room temperature and shaken out once with 2N hydrochloric acid and once with a 5% sodium carbonate solution. The organic phase is separated and dried on magnesium sulfate. After concentration by evaporation in a vacuum, the residue is recrystallized from methyl-tert-butyl ether. 6.06 g of waxy flakes is obtained.
Yield: 91%
Cld: C 68.01% H 7.58% N 4.10% S 6.26%
Fnd: C 67.85% H 7.43% N 3.98% S 6.17%

Example 22 a) 7-N-~tert-Butoxycarbonyl)-2-(4-benzyloxybenzyl)-1-tosyl-1,4,7-triazaheptan-3-one 20 g (42.77 mmol) of the title compound of Example 16b) is dissolved in 200 ml of chloroform, and 4.76 g (47.05 mmol) of triethylamine is added. At 0~C, a solution of 10.27 g (47.05 mmol) of di-tert-butyl dicarbonate in 50 ml of chloroform is added in drops and stirred for 30 minutes at 0~C. Then, it is stirred for 5 hours at room temperature. It is shaken out three times with 5% sodium carbonate solution, the organic phase is dried on magnesium sulfate and concentrated by evaporation in a vacuum. The residue is recrystallized from a little methanol.
22.34 g of colorless crystals is obtained.
Yield: 92%
Cld: C 63.47% H 6.57~ N 7.40% S 5.65%
Fnd: C 63.31% H 6.42% N 7.45% S 5.49%

b) 7-N-tert-Butoxycarbonyl-2-(4-hydroxybenzyl)-1-tosyl-1,4,7-triazaheptan-3-one 21 g (36.99 mmol) of the title compound of Example 22a) is dissolved in 300 ml of methylene chloride and 4 g of palladium catalyst (10% Pd/C) is added. It is hydrogenated overnight. The catalyst is filtered out, and the filtrate is evaporated to dryness. 16.39 g of a vitreous foam is obtained, which solidifies after a short time.
Yield: 99%
Cld: C 57.84% H 6.54% N 8.80% S 6.71%
Fnd: C 57.70% H 6.61% N 8.69% S 6.54%

c) 7-N-tert-Butoxycarbonyl-2-t4-(benzyloxycarbonylmethyloxy)-benzyl]-1-tosyl-1,4,7-triazaheptan-3-one 15 g (33.51 mmol) of the title compound of Example 22b), 7.68 g (33.51 mmol) of bromoacetic acid benzyl ester and 13.8 g (100 mmol) of potassium carbonate are refluxed in 300 ml of acetonitrile for 24 hours. The salts are filtered out, and the filtrate is evaporated to dryness. The residue is dissolved in 200 ml of methylene chloride and shaken out twice with 100 ml of water. The organic phase is dried on magnesium sulfate and concentrated by evaporation in a vacuum. The residue is chromatographed on silica gel (mobile solvent: methylene chloride/hexane/acetone: 20/10/1). 10.69 g of a colorless oil is obtained.
Yield: 51%
Cld: C 61.42% H 6.28~ N 6.72% S 5.12%
Fnd: C 61.27% H 6.09% N 6.68% S 5.03%

d) 1-Tosyl-2-(4-~benzyloxycarbonylmethyloxy)-benzyl)-1,4,7-triazaheptan-3-one (a~ trifluoroacetate salt) 10 g (15.98 mmol) of the title compound of Example 22c) is stirred for 1 hour in 100 ml of trifluoroacetic acid at room temperature. It is evaporated to dryness in a vacuum. 10.22 g of a vitreous foam is obtained, which solidifies with standing.
Yield: 100%
Cld: C 54.45% H 5.04% N 6.57% S 5.01% F 8.91%
Fnd: C 54.51% H 5.10% N 6.43% S 4.89% F 9.15%

e) 9-Chloro-2-~4-(benzyloxycarbonylmethyloxy)-benzyl)-1-tosyl-1,4,7-triazanonane-3,8-dione 10 g (15.63 mmol) of the title compound of Example 22d) and 4.75 g (46.90 mmol) of triethylamine are dissolved in 200 ml of chloroform. At 0~C, 1.94 g (17.19 mmol) of chloroacetyl chloride is added in drops within 30 minutes and then stirred for 2 hours at 0~C. The organic phase is shaken out twice with 5%
hydrochloric acid and twice with water, dried on magnesium sulfate and evaporated to dryness in a vacuum. The residue is ~ ~ q456 1 chromatographed on silica gel (mobile solvent: methylene chloride/ethyl acetate = 20:1). 7.62 g of a waxy solid is obtained.
Yield: 81%
Cld: C 57.85% H 5.36% N 6.98% S 5.32% Cl 5.89%
Fnd: C 57.70% H 5.49% N 6.82% S 5.25% Cl 5.78%

f) 9-Chloro-2-(4-(carboxymethyloxy)-bensyl)-1-tosyl-1,4,7-triazanonane-3,8-dione 7 g (11.63 mmol) of the title compound of Example 22e) is dissolved in 150 ml of methylene chloride and mixed with 2 g of palladium catalyst (10% Pd/C). It is hydrogenated overnight.
The catalyst is filtered out, and the filtrate is evaporated to dryness in a vacuum. 5.89 g of a vitreous solid is obtained.
Yield: 99%
Cld: C 51.61% H 5.12% N 8.21% S 6.26% Cl 6.92%
Fnd: C 51.45% H 5.03% N 8.13% S 6.11% Cl 6.79%

g) 10-Acetyl-2-(4-(carboxymethyloxy)-benzyl)-1-tosyl-10-thia-1,4,7-triazadecane-3,8-dione 5 g (9.77 mmol) of the title compound of Example 22f) is dissolved in 80 ml of chloroform and 1.98 g (19.53 mmol) of triethylamine is added. 0.74 g (9.77 mmol) of thioacetic acid is added and refluxed for lo minutes. The solution is poured into 200 ml of ice-cooled 5% hydrochloric acid and stirred vigorously.
The organic phase is separated, dried on magnesium sulfate and evaporated to dryness in a vacuum. Chromatographic purification on silica gel (mobile solvent: hexane/ethyl acetate = 3:1) yields 4.47 g of the title compound as vitreous solid.
Yield: 83%
Cld: C 52.26% H 5.30% N 7.62% S 11.62%
Fnd: C 52.11% H 5.39% N 7.50% S 11.49%

h) N-Hydroxysuccinimide~ter of 10-acetyl-2-(4-~carboxymethyloxy)-benzyl)-l-tosyl-10-thia-1,4,7-triazadecane-3,8-dione 4 g (7.25 mmol) of the title compound of Example 22g), 1.65 g (7.98 mmol) of dicyclohexylcarbodiimide, 30 mg of 4-dimethylaminopyridine and 0.92 g (7.98 mmol) of N-hydroxysuccinimide are dissolved at 0~C in 20 ml of chloroform and stirred for 1 hour at this temperature. Then, stirring is continued for 24 hours at room temperature. 20 ml of ether is added, the precipitated dicyclohexylurea is suctioned off, and the filtrate is concentrated by evaporation in a vacuum. The residue is chromatographed on silica gel (mobile solvent:
methylene chloride/dioxane = 10:1). 4.09 g of a colorless solid is obtained.
Yield: 87%
Cld: C 51.84% H 4.97~ N 8.64~ S 9.88%
Fnd: C 51.68% H 4.80% N 8.53% S 9.68%

Example 23 a) 4-Nitrophenolester of 10-acetyl-2-(4-(carboxymethyloxy)-benzyl)-1-tosyl-10-thia-1,4,7-triazadecane-3,8-dione 4 g (7.25 mmol) of the title compound of Example 22g), 1.11 g (7.97 mmol) of 4-nitrophenol, 30 mg of 4-dimethylaminopyridine and 1.65 g (7.97 mmol) of dicyclohexylcarbodiimide are dissolved at 0~C in 20 ml of chloroform and stirred for 3 hours at this temperature. Then, it is stirred for 24 hours at room temperature. 20 ml of ether is added, settled precipitate is suctioned out, and the filtrate is concentrated by evaporation in a vacuum. The residue is chromatographed on silica gel (mobile solvent: methylene chloride/dioxane = 15:1). 3.85 g of a cream-colored solid is obtained.
Yield: 79%
Cld: C 53.56% H 4.79% N 8.33% S 9.53%
Fnd: C 53.41% H 4.63% N 8.17% S 9.38%

Example 24 a) Pentafluorophenolester of 10-acètyl-2-(4-(carboxymethyloxy)-benzyl)-1-tosyl-10-thia-1,4,7-triazadecane-3,8-dione 4 g (7.25 mmol) of the title compound of Example 22g), 1.47 g (7.97 mmol) of pentafluorophenol, 30 mg of 4-dimethylaminopyridine and 1.65 g (7.97 mmol) of dicyclohexylcarbodiimide are dissolved at 0~C in 20 ml of chloroform and stirred for 3 hours at this temperature. Then, it is stirred for 24 hours at room temperature. 20 ml of ether is added, settled precipitate is suctioned out, and the filtrate is 77 2l ~ 456l concentrated by evaporation in a vacuum. The residue is chromatographed on silica gel (mobile solvent: methylene chloride/dioxane = 15:1). 3.95 g of a colorless solid is obtained.
Yield: 76%
Cld: C 50.20% H 3.93% N 5.85% S 8.93% F 13.24%
Fnd: C 50.05% H 3.87% N 5.69% S 8.71% F 13.03%

Example 25 a) 7-N-tert-Butoxycarbonyl-2-(4-benzyloxybenzyl)-1-me~yl-1,4,7-triazaheptan-3-one 16.23 g (42.77 mmol) of the title compound of Example l9b) is dissolved in 200 ml of chloroform and 4.76 g (47.05 mmol) of triethylamine is added. At 0~C, a solution of 10.27 g (47.05 mmol) of di-tert-butyldicarbonate in 50 ml of chloroform is added in drops and stirred for 30 minutes at 0~C. Then, it is stirred for 5 hours at room temperature. It is shaken out three times with 5% sodium carbonate solution, the organic phase is dried on magnesium sulfate and concentrated by evaporation in a vacuum.
The residue is recrystallized from a little methanol. 20.19 g of a foamy solid is obtained.
Yield: 96%
Cld: C 58.64% H 6.77% N 8.55% S 6.52%
Fnd: c 58.48% H 6.59% N 8.41% S 6.42 78 21~4561 b) 7-N-tert-Butoxycarbonyl-2-(4-hydroxybenzyl)-l-me5yl-1,4,7-triazaheptan-3-one 20 g (40.68 mmol) of the title compound of Example 25a) is dissolved in 300 ml of methylene chloride, and 4 g of palladium catalyst (10% Pd/C) is added. It is hydrogenated overnight. The catalyst is filtered out, and the filtrate is evaporated to dryness. 16.17 g of a vitreous foam is obtained, which solidifies after a short time.
Yield: 99%
Cld: C 50.86% H 6.78% N 10.47% S 7.99%
Fnd: C 50.70% H 6.69% N 10.31% S 7.78%

c) 7-N-tert-Butoxycarbonyl-2-(4-benzyloxycarbonylmethyloxy)-benzyl)-l-mesyl-1,4,7-triazaheptan-3-one 15 g (37.36 mmol) of the title compound of Example 25b), 8.56 g (37.36 mmol) of bromoacetic acid benzyl ester and 13.8 g (100 mmol) of potassium carbonate are refluxed in 300 ml of acetonitrile for 24 hours. The salts are filtered out, and the filtrate is evaporated to dryness. The residue is dissolved in 200 ml of methylene chloride and shaken out twice with 100 ml of water. The organic phase is dried on magnesium sulfate and concentrated by evaporation in a vacuum. The residue is chromatographed on silica gel (mobile solvent: methylene chloride/hexane/acetone = 20/10/1). 10.06 g of foamy solid is obtained.
Yield: 49%

Cld: C 56.82~ H 6.42% N 7.64% S 5.83%
Fnd: C 56.65% H 6.35% N 7.51% S 5.72%

d) 2-(4-(Benzyloxycarbonylmethyloxy)-benzyl)-1-mesyl-1,4,7-triazaheptan-3-one (as trifluoroacetic acid salt) 10 g (18.19 mmol) of the title compound of Example 25c) is stirred for 1 hour in 100 ml of trifluoroacetic acid at room temperature. It is evaporated to dryness in a vacuum. 9.95 g of a vitreous foam is obtained, which solidifies in the case of standing.
Yield: 97%
Cld: C 49.02% H 5.01% N 7.46% S 5.69% F 10.11%
Fnd: C 48.91% H 4.90% N 7.30% S 5.51% F 9.96%

e) 9-Chloro-2-(4-(benzyloxycarbonylmethyloxy)-benzyl)-1-mesyl-1,4,7-triazanonane-3,8-dione 9 g (15.97 mmol) of the title compound of Example 25d), 1.78 g (17.57 mmol) of triethylamine are dissolved in 200 ml of chloroform. At 0~C, 1.98 g (17.57 mmol) of chloroacetyl chloride is added in drops within 30 minutes and then stirred for 2 hours at 0~C. The organic phase is shaken out twice with 5%
hydrochloric acid and twice with water, dried on magnesium sulfate and evaporated to dryness in a vacuum. The residue is chromatographed on silica gel (mobile solvent: methylene chloride/ethyl acetate = 20:1).
Yield: 83%

- - -Cld: C 52.52% H 5.37% N 7.99% S 6.09% Cl 6.74%
Fnd: C 52.37% H 5.43 N 7.81 S 5.93% Cl 6.58%

f) 9-Chloro-2-(4-(carboxymethyloxy)-benzyl)-1-mesyl-1,4,7-triazanonane-3,8-dione 6.5 g (12.36 mmol) of the title compound of Example 25e) is dissolved in 150 ml of methylene chloride and mixed with 2 g of palladium catalyst (10% Pd/C). It is hydrogenated overnight.
The catalyst is filtered out, and the filtrate is evaporated to dryness in a vacuum. 5.33 g of a vitreous solid is obtained.
Yield: 99%
Cld: C 44.09% H 5.09% N 9.64% S 7.36% Cl 8.13%
Fnd: C 43.93% H 4.95% N 9.52% S 7.22% Cl 8.03%

g) 10-Acetyl-2-(4-(carboxymethyloxy)-benzyl)-1-mesyl-10-thia-1,4~7-triazadecane-3,8-dione 5 g (11.47 mmol) of the title compound of Example 22f) is dissolved in 80 ml of chloroform, and 1.98 g (19.53 mmol) of triethylamine is added. 0.74 g (9.77 mmol) of thioacetic acid is added and refluxed for 10 minutes. The solution is poured into 200 ml of ice-cooled 5% hydrochloric acid and stirred vigorously.
The organic phase is separated, dried on magnesium sulfate and evaporated to dryness in a vacuum. Chromatographic purification on silica gel (mobile solvent: hexanetethyl acetate = 3;1) yields 4.64 g of the title compound as vitreous solid.

Yield: 85%

Cld: C 45.46% H 5.30% N 8.84% S 13.48%
Fnd: C 45.28% H 5.17% N 8.61% S 13.38%

h) N-Hydroxysuccinimidester of 10-acetyl-2-(4-(carboxymethyloxy)-benzyl)-1-mesyl-10-thia-1,4,7-triazadecane-3,8-dione 4 g (8.41 mmol), 4 g (7.25 mmol) of the title compound of Example 25g), 1.91 g (9.25 mmol) of dicyclohexylcarbodiimide, 30 mg of 4-dimethylaminopyridine and 1.06 g (9.25 mmol) of N-hydroxysuccinimide are dissolved at 0~C in 20 ml of chloroform and stirred for 1 hour at this temperature. Then, stirring is continued for 24 hours at room temperature. 20 ml of ether is added, precipitated dicyclohexylurea is suctioned out, and the filtrate is concentrated by evaporation in a vacuum. The residue is chromatographed on silica gel (mobile solvent: methylene chloride/dioxane = 10:1). 3.47 g of a cream-colored solid is obtained.
Yield: 72%
Cld: C 46.15% H 4.93% N 9.78% S 11.20%
Fnd: C 46.03% H 4.83% N 9.64% S 11.05%

Example 26 a) 3-Glycerol-ester of 10-acetyl-2-(4-(carboxymethyloxy)-benzyl)-1-mesyl-10-thia-1,4,7-triazadecane-3,8-dione with glycerol-1,2-dipalmitate 2 g (5.27 mmol) of the title compound of Example 25g), 20 mg of 4-dimethylaminopyridine, 3.30 g (5.80 mmol) of glycerol-1,2-82 21 q4561 dipalmitic acid ester and 1.20 g (5.80 mmol) ofdicyclohexylcarbodiimide are dissolved at 0~C in 5 ml of chloroform and stirred for 3 hours at this temperature. Then, it is stirred for 24 hours at room temperature. 20 ml of ether is added, and precipitate is filtered out. The filtrate is evaporated to dryness in a vacuum, and the residue is chromatographed on silica gel (mobile solvent: hexane/ethyl acetate = 30:1). 3.35 g of a waxy solid is obtained.
Yield: 62%
Cld: C 62.02% H 8.94% N 4.09% S 6.25%
Fnd: C 61.91% H 8.75% N 3.91% S 6.18%

Example 27 a) 4-Benzyl ether of N-mesyl-tyrosine 10 g (27.5 mmol) of the title compound of Example l9a) and 5.50 g (137.6 mmol) of sodium hydroxide are refluxed in a mixture of 50 ml of water/150 ml of ethanol for 3 hours. It is evaporated to dryness, the residue is taken up in 200 ml of 3N
hydrochloric acid, and it is stirred for 2 hours at room temperature. The precipitated acid is suctioned off, washed with water and dried in a vacuum at 70~C. 9.42 g of cream-colored solid is obtained.
Yield: 98%
Cld: C 58.44% H 5.48% N 4.01% S 9.18%
Fnd: C 58.28% H 5.37% N 3.91% S 9.02%

b) 6~6~-Bis-tl-N-tert-butoxycarbony~ 4-diaza-hexan-3-one]
disulfide 10 g (65.67 mmol) of cystamine, 11.50 g (65.67 mmol) of N-boc-glycine and 14.90 g (72.24 mmol) of dicyclohexylcarbodiimide are dissolved at 0~C in 50 ml of tetrahydrofuran and stirred for 2 hours at this temperature. Then, it is stirred for 12 hours at room temperature. 50 ml of ether is added, settled precipitate is suctioned out, and the filtrate is evaporated to dryness. The residue is chromatographed on silica gel (mobile solvent:
hexane/acetone = 6:1). 20.84 g of a vitreous solid is obtained.
Yield: 68%
Cld: C 46.33% H 7.34% N 12.01% S 13.74%
Fnd: C 46.15% H 7.28% N 11.93% S 13.67%

c) 6,6'-Bi~-[1,4-diaza-hexan-3-one]-disulfide 20 g (42.86 mmol) of the title compound of Example 27b) is dissolved in 100 ml of trifluoroacetic acid, and it is stirred for 2 hours at room temperature. It is evaporated to dryness, the residue is taken up with 300 ml of 10% sodium carbonate solution and extracted 6 times with 50 ml of chloroform. The combined chloroform phases are dried on magnesium sulfate and concentrated by evaporation in a vacuum. 10.96 g of a slightly yellow-colored solid is obtained.

Yield: 969~
Cld: C 36.07% H 6.81% N 21.03% S 24.07%

Fnd: C 35.91% H 6.90% N 20.89% S 23.89%

d) 9,9'-Bis[2-(4-benzyloxybenzyl)-1-mesyl-1,4,7-triazanonane-3,6-dione]disulfide 9 g (25.76 mmol) of the title compound of Example 27a), 3.43 g (12.87 mmol) of the title compound of Example 27c) and 6.19 g (30 mmol) of dicyclohexylcarbodiimide are dissolved at 0~C
in 40 ml of tetrahydrofuran and stirred for 2 hours at this temperature. Then, it is stirred for 12 hours at room temperature. 30 ml of ether is added, and precipitate is suctioned out. The filtrate is evaporated to dryness in a vacuum, and the residue is chromatographed on silica gel (mobile solvent: methylene chloride/acetone = 20:1). 5.59 g of a cream-colored solid is obtained.
Yield: 48% [relative to 27c)]
Cld: C 53.08% H 5.79% N 9.28% S 14.17%
Fnd: C 52.93% H 5.84% N 9.13 S 14.02%

Example 28 a) 4-Benzyl ether of N-mesyl-tyrosinamide 20 g (55.03 mmol) of the title compound of Example l9a) is dissolved in 300 ml of tetrahydrofuran, and an ammonia stream is introduced at 0~C for 3 hours. It is evaporated to dryness, and the residue is recrystallized from methanol. 18.6 g of colorless flakes is obtained.
Yield; 97~
Cld: C 58.60% H 5.79% N 8.04% S 9.20%

Fnd: C 58.47% H 5.88% N 7.91% S 9.05%

2 1 ~456 1 b) 2-~4-Benzyloxybenzyl)-1-mesyl-1,4-diazabutane 18 g (51.66 mmol) of the title compound of Example 28a) is dissolved in 200 ml of tetrahydrofuran, and 310 ml of 1 M
diborane in THF is added under nitrogen atmosphere. It is refluxed for 24 hours. It is cooled to 0~C in an ice bath, and 70 ml of concentrated hydrochloric acid is added. Then, it is refluxed for 5 hours. It is evaporated to dryness, and the residue is taken up with 300 ml of saturated sodium carbonate solution. It is extracted 3 times with 100 ml of methylene chloride, the combined phases are dried on magnesium sulfate and evaporated to dryness in a vacuum. The chromatographic purification is carried out on silica gel (mobile solvent:
methylene chloride/ethanol = 10:1). 15.38 g of a cream-colored solid is obtained.
Yield: 89%
Cld: C 61.05% H 6.63% N 8.38% S 9.59%
Fnd: C 60.91% H 6.54% N 8.27% S 9.41%

c) 4-N-Butoxycarbonyl-2-(4-benzyloxybenzyl)-1-me~yl-1,4,7-triazaheptan-5-one 15 g (44.85 mmol) of the title compound of Example 28b) is dissolved in 200 ml of chloroform, and 5 g (49.34 mmol) of triethylamine and 12.84 g (49.34 mmol) of N-tert-butoxycarbonyl-glycine-N-hydroxysuccinimide ester are added at 0~C. It is stirred for 12 hours at room temperature. It is extracted twice with cold 5% hydrochloric acid and once with water. The organic phase is dried on magnesium sulfate and evaporated to dryness in a vacuum. 20.51 g of an amorphous solid is obtained.
Yield: 93%
Cld: C 58.64% H 6.77% N 8.55% S 6.52%
Fnd: C 58.47% H 6.85% N 8.43% S 6.41%

d) 2-(4-Benzyloxybenzyl)-1-mesyl-1,4,7-triazaheptan-5-one 20 g (40.68 mmol) of the title compound of Example 28c) is dissolved in 100 ml of trifluoroacetic acid and dissolved for 5 hours at room temperature. It is evaporated to dryness in a vacuum, the residue is taken up in 200 ml of saturated sodium carbonate solution and extracted 3 times with 100 ml of chloroform. The organic phases are dried on sodium sulfate and concentrated by evaporation in a vacuum. 15.61 g of a vitreous solid is obtained.
Yield: 98%
Cld: C 58.29% H 6.44% N 10.73% S 8.19%
Fnd: C 58.13% H 6.60~ N 10.61% S 8.05%

e) 9-Chloro-2-(4-benzyloxybenzyl)-1-mesyl-1,4,7-triazanonane-5,8-dione 10 g (25.54 mmol) of the title compound of Example 28d) and 2.58 g (25.54 mmol) of triethylamine are dissolved in 200 ml of chloroform, 2.88 g (25.54 mmol) of chloroacetyl chloride is added in drops at 0~C within 30 minutes. It is stirred for 3 hours at 0~C. It is poured onto 200 ml of 5% cold hydrochloric acid and stirred vigorously. The organic phase is separated, -' ' 21~4561 dried on magnesium and evaporated to dryness in a vacuum. The residue is recrystallized from methanol. 11.36 g of a cream-colored solid is obtained.
Yield: 95%
Cld: C 53.90% H 5.60% N 8.98~ S 6.85% Cl 7.58~
Fnd: C 53.80% H 5.71% N 8.91% S 6.73% Cl 7.44%

f) 10-Benzoyl-2-~4-benzyloxybenzyl)-1-mesyl-1,4,7-triaza-10-thiadecane-5,8-dione 5 g (10.68 mmol) of the title compound of Example 28e), 1.08 g (10.68 mmol) of triethylamine and 1.48 g (10.68 mmol) of thiobenzoic acid are refluxed in 50 ml of chloroform for 10 minutes. It is evaporated to dryness, and the residue is chromatographed on silica gel (mobile solvent: methylene chloride/acetone = 15:1). 4.93 g of a cream-colored, amorphous solid is obtained.
Yield: 81%
Cld: C 59.03% H 5.48% N 7.38% S 11.26%
Fnd: C 58.87% H 5.31% N 7.25% S 11.04%

Example 29 a) 9,9'-Bis-[2-~4-benzyloxybenzyl)-1-mesyl-1,4,7-triazanonan-3-one]-disulfide 2 00 g (13.17 mmol) of 1,6-dichloro-3,4-dithiahexane (dissolved in 20 ml of acetonitrile) is added in drops to 10 g (26.35 mmol) of the title compound of Example l9b) and 10.92 g (79 mmol) of potassium carbonate in 100 ml of acetonitrile in the boiling heat within 1 hour. It is refluxed for 12 hours. The salts are filtered out, the filtrate is evaporated to dryness in a vacuum, and the residue is chromatographed on silica gel (mobile solvent: methylene chloride/isopropanol = 10:1). 2.19 g of a slightly yellow-colored crystalline powder is obtained.
Yield: 19% (relative to 1,6-dichloro-3,4-dithiahexane) Cld: C 54.77% H 6.43% N 9.58% S 14.62%
Fnd: C 54.61% H 6.53% N 9.41% S 14.51%

Example 30 a) 6-Chloro-2-(4-benzyloxybenzyl)-1-mesyl-1,4-diazahexan-5-one 10 g (25.54 mmol) of the title compound of Example 28b) is dissolved together with 3.03 g (29.90 mmol) of triethylamine in 200 ml of chloroform. 3.38 g (29.90 mol) of chloroacetyl chloride is added in drops at 0~C, and it is stirred for 2 hours at this temperature. It is poured onto 200 ml of 5% cold hydrochloric acid and stirred well. The organic phase is separated, dried on magnesium sulfate and evaporated to dryness in a vacuum. The residue is recrystallized from methanol. 11.67 g of colorless crystals is obtained.
Yield: 95%
Cld: C 55.54% H 5.64% N 6.82% S 7.80% Cl 8.63%
Fnd: C 55.38% H 5.71% N 6.67% S 7.63% Cl 8.51%

b) 9,9'-Bis-[2-(4-benzyloxybenZyl)-l-mesyl-1,4,7-triaZanonan-5-one]-disulfide 10 g (24.34 mmol) of the title compound of Example 30a), 1.52 g (10 mmol) of cystamine and 8.29 g (60 mol) of potassium carbonate are refluxed in 150 ml of tetrahydrofuran for 8 hours.
The salts are filtered out, the filtrate is evaporated to dryness in a vacuum, and the residue is chromatographed on silica gel (mobile solvent: methylene chloride/ethanol = 15:1). 2.02 g of a slightly yellowish solid is obtained.
Yield: 23% (relative to cystamine) Cld: C 54.77% H 6.43% N 9.58% S 14.62%
Fnd: C 54.61% H 6.52% N 9.47% S 14.48%

Example 31 a) Cystamine-bis-~chloroacetamide) 10 g (65.67 mmol) of cystamine and 13.29 g (131.34 mmol) of triethylamine are dissolved in 100 ml of chloroform at 0~C.
14.38 g (131.34 mol) of chloroacetyl chloride is added in drops, and it is stirred for 3 hours at 0~C. The solution is poured into 200 ml of cold 5% hydrochloric acid and stirred vigorously.
The organic phase is dried on magnesium sulfate and concentrated by evaporation in a vacuum. The residue is recrystallized from a little acetone. 17.04 g of a cream-colored solid is obtained.
Yield: 85%
Cld: C 31.48% H 4.62% N 9.18% S 21.01% Cl 23.23%
Fnd: c 31.27% H 4.51% N 9.09% S 20.93% Cl 23.12%

-b) 9,9'-Bist2-~4-benzyloxybenzyl)-1-mesyl-1,4,7-triazanonan-6-one]-di~ulfide 5 g (16.38 mmol) of the title compound of Example 31a), 13.69 g (40.95 mmol) of the title compound of Example 28b) and 20.73 g (150 mmol) of potassium carbonate are refluxed in 200 ml of ethanol for 8 hours. 200 ml of methylene chloride is added, salts are suctioned out, and the filtrate is concentrated by evaporation in a vacuum. The residue is chromatographed on silica gel (mobile solvent: methylene chloride/ethanol = 15:1).
4.74 g of a vitreous solid is obtained.
Yield: 33 Cld: C 54.77% H 6.43% N 9.58% S 14.62~
Fnd: C 54.65% H 6.37% N 9.41~ S 14.53%

Example 32 a) 2-t4-Benzyloxybenzyl]-l-me~yl-1,4,7-triazaheptane 5 g (13.18 mmol) of the title compound of Example l9b) is dissolved in 50 ml of tetrahydrofuran, and 80 ml of 1 M diborane solution (1 M in THF) is added. It is refluxed for 24 hours. It is cooled to 0~C, and 20 ml of concentrated hydrochloric acid is added. Then, it is refluxed for 5 hours. The solution is evaporated to dryness and taken up with 200 ml of saturated sodium carbonate solution. Then, it is extracted 5 times with loo ml of chloroform. The combined organic phases are dried on sodium sulfate and concentrated by evaporation in a vacuum. The residue is chromatographed on silica gel (mobile solvent:

methylene chloride/isopropanol = 8:1, +2% NH40H). 4.19 g of a vitreous solid is obtained.
Yield: 87%
Cld: C 59.15% H 7.45% N 11.50% S 8.77%
Fnd: C 59.03% H 7.28% N 11.37% S 8.61%

b) 9,9'-Bi~-t2-~4-benzyloxybenzyl)-1-mesyl-1,4,7-triazanonan-8-one]-disulfide 4 g (10.94 mmol) of the title compound of Example 32a), 0.67 g (3.65 mmol) of 2,2'-dithiodiacetic acid and 2.48 g (12.04 mmol) of dicyclohexylcarbodiimide are dissolved at 0~C in 20 ml of tetrahydrofuran and stirred for 3 hours at this temperature.
Then, it is stirred for 24 hours at room temperature. 20 ml of ether is added, precipitate is filtered out, and the filtrate is concentrated by evaporation in a vacuum. The residue is chromatographed on silica gel (mobile solvent: methylene chloride/ethanol = 15:1). 0.99 g of an amorphous solid is obtained.
Yield: 31% (relative to dicarboxylic acid) Cld: C 54.77% H 6.43% N 9.58% S 14.62%
Fnd: C 54.58% H 6.38% N 9.47% S 14.51%

Claims (12)

Claims

1. Compounds of general formula I

in which V1, V2, V3, V4, independently of one another, stand for a carbonyl, >CH(COOH) or -CH2 group, X1 stands for a hydrogen atom, a C1-C12 alkyl radical optionally substituted with a carboxyl, an amino or a thiocyanate group or a metal ion equivalent of a radioactive metal ion of an element of atomic number 43, 45, 46, 75, 82 or 83, X2, X3, independently of one another, stand for a hydrogen atom or a metal ion equivalent of a radioactive metal ion of an element of the above-mentioned atomic numbers, n, m, p stand for numbers 0 or 1, whereby m + n = 1 holds true, R1 stands for a hydrogen atom, a carboxyl group or a group -U-Z, in which U stands for a direct bond, a straight-chain or branched, saturated or unsaturated C1-C20 alkylene radical, which optionally contains a maleimide radical, a succinimide radical, a phenyl radical optionally substituted by 1 to 5 fluorine atoms, an amino or nitro group, one or two imino, phenylene, phenylenoxy, phenylenamino, amide, hydrazide, carbonyl, ureido, thioureido, thioamide, ester group(s), 1 to 2 oxygen, sulfur and/or nitrogen atom(s) as well as optionally 1 to 5 hydroxy, mercapto, oxo, thioxo, carboxy, alkylcarboxylic acid, ester, thiocyanate and/or amino groups, and Z stands for a hydrogen atom, a radical of an amino acid, a peptide, a polynucleotide or a steroid or a functional group with which optionally the radical of an amino acid, a peptide, a polynucleotide or a steroid is bound, R2 stands for a straight-chain or branched C1-C10 alkyl radical, which optionally contains a -COOH group, a C7-C12 aralkyl radical or an aromatic compound, which optionally is substituted with a chlorine or bromine atom, a thiocyanate, a methyl, ethyl, carboxyl and/or methoxy group, R4 stands for a hydrogen atom or a carboxyl group or if R1 means a hydrogen atom or a carboxyl group, it stands in addition for a group -U-Z, in which U and Z have the indicated meanings, R3 stands for a hydrogen atom, a metal ion equivalent of an element of the above-mentioned atomic numbers, a trifluoroacetate, acetate, benzoate, C1-C6 acyl, a benzoyl, a hydroxyacetyl, an acetamidomethyl radical, a benzoic acid radical optionally substituted with a chlorine or bromine atom, a methyl, ethyl, carboxyl and/or methoxy group, a p-methoxybenzyl radical, an ethoxyethyl radical, an SH protective group, a radical, or if X2, X3 stand for a hydrogen and X1 stands for a hydrogen or an optionally substituted C1-C12 alkyl radical, for a radical of formula II

in which V1, V2, V3, V4, X1, X2, X3, n, m, p, R1, R2 and R4 have the indicated meanings, whereby at least one and at most two radicals V1, V2, V3, V4 stand for a carbonyl group.

2. Compounds of general formula I, characterized in that at least two of radicals X1, X2, X3 or R3 stand for a metal ion equivalent of a radioactive metal isotope of atomic numbers 43, 45, 46, 75, 82 or 83.

3. Compound according to one of the preceding claims, wherein Z stands for an amino acid or a peptide.

4. Compound according to one of the preceding claims, in which Z stands for a polynucleotide.

5. Compound according to one of the preceding claims, wherein V1 and V4 each stand for a carbonyl group, V2 and V3 each for a -CH2 group and p for number 0.

6. Compound according to one of the preceding claims, wherein a 99m-technetium is contained as a radioactive metal ion.

7. Compound according to one of the preceding claims, wherein R4 is hydrogen or a carboxylic acid group.

8. Compound according to one of the preceding claims, wherein R2 is a p-CH3-C6H4 radical.

9. Compound according to claim 1, wherein X1, X2 and X3 stand for a hydrogen atom and R3 stands for a radical of formula II.

10. Pharmaceutical agents, containing at least one metal complex of formula I, in which at least two of radicals X1, X2, X3 and/or R3 stand for a metal ion equivalent.

11. Use of a metal complex according to claim 1 in radiodiagnosis or radiotherapy.

12. Process for the production of a pharmaceutical agent according to claim 10, wherein a compound of formula I with X2 and X3 meaning a hydrogen atom and X1 meaning hydrogen or an optionally substituted C1-C12 alkyl radical and a reducing agent is dissolved in aqueous medium with the addition of the additives usual in galenicals and then, optionally with the addition of a transfer ligand, is reacted with a metal salt or metal oxide of the desired metal ion, and optionally is mixed with a pharmacologically acceptable radiological vehicle, and the complexing agent is added in excess, optionally in the form of its alkali salt.