WO1994022491A1 - Bifunctional chelators and their use in radiopharmaceuticals - Google Patents

Abstract

New technetium and rhenium chelate compounds are disclosed, as well as a process for preparing the same, radiopharmaceuticals containing these compounds, conjugates of these compounds with substances which selectively accumulate in diseased tissues, in particular peptides and proteins, as well as the preparation of compositions containing these compounds and their use in radiodiagnostic examinations.

Description

BIFUNCTIONAL CHELATORS AND THEIR USE IN RADIOPHARMACEUTICAL.

The present invention relates to new technetium and rhenium chelate compounds, processes for their preparation and radiopharmaceutical compositions containing these compounds, and conjugates of these compounds with selectively enriching themselves in diseased tissue

Substances, especially peptides and proteins. The invention further relates to the production of compositions containing these compounds and their use for radiodiagnostic examinations.

Radioactive metal ions have long been used in medical diagnostics and therapy. For example, gamma-ray emitters such as the isotope Tc-99m are used for tumor detection. ß-emitters, such as the isotopes Re-186, Re-188 and Re-189 are found in the

Tumor therapy application. The most frequently used radionuclide for nuclear medicine questions is technetium-99m, which due to its favorable physical properties (no corpuscular radiation, 6 h physical half-life, 140 keV gamma radiation) and the resulting low radiation exposure are particularly good as radioisotopes for which is suitable for in vivo diagnostics. Technetium-99m can easily be obtained from nuclide generators as pertechnetate and can be used directly in this form for the production of kits for routine clinical needs.

The efficiency of radionuclides in in vivo diagnostics as well as therapy depends on the specificity and the selectivity of the labeled chelates to the target cell. These properties can be improved by coupling the chelates to biomolecules according to the "drug targeting" principle. Antibodies, their fragments, hormones, growth factors and substrates of receptors and enzymes are suitable biomolecules. The British patent application GB 2,109,407 describes the use of radioactively labeled monoclonal antibodies against tumor-associated antigens for in vivo tumor diagnosis. Likewise, direct protein labels via donor groups (amino, amide, thiol, etc.) of the protein (Rhodes, BA et al., J. Nukl. Med. 1986, 27, 685-693) or by introducing complexing agents (US Pat. No. 4,479,930 and Fritzberg, AR et al., J. Nucl. Med. 1986, 27, 957) with Techne¬ tium-99m. However, these experimental methods are not available for clinical use because on the one hand the selectivity is too low and on the other hand the background activity in the organism is too high to enable in vivo imaging.

The first receptor-binding radiopharmaceuticals were published by JP DiZio et al. (J. Nucl. Ed. 1992, 33; 558-569 and Bioconjugate Chem. 1991, 2; 353-366). In vivo imaging could not be achieved with these compounds. The necessity of extracting the described compounds from the labeling medium proves to be a great disadvantage. These conditions are not satisfactory, since on the one hand the preparation of the kit for the user has to be carried out with the lowest possible radiation exposure and on the other hand suitable laboratories are available in only a few clinics for carrying out this work. Cyclame as described by Volkert et al. (Appl.Radiol.Isot. 1982, 33; 891-896) and Troutner et al. (J.Nucl. Ed. 1980, 21; 443-448), as well as N ₂ S ₂ and N3S systems, because of the lack of reactive groups for linking to biomolecules or proteins per se, have little use as tissue-specific radiopharmaceuticals (M. Borel et al., Appl. Radiat. Isot. 1992, 43, 425-436).

While for the labeling of the cyclame mentioned above and also known N ₂ 0 ₂ systems (Pillai, MRA et al .; Inorg. Chem. 1990, 29, 1850-1856) pH values of 10-13 are required, the Rule N ₂ S ₂ and N ₃ S systems already at pH values of 9-11. These conditions are still too extreme for labeling labile conjugates of bio and macromolecules, especially since some systems, such as L, L-ethylenedicysteine (L, L-EC), only change at pH values of> 10 (Verbruggen, AM et al .; J. Nucl. Med. 1992, 33, 551-557) have sufficient markings. The known MAG3 even requires a temperature influence of 90-100 ° C for 10 minutes (Bannister, KM et al., - J. Nucl. Med. 1990, 31, 1568-1573) in order to have a radiochemical sufficient for clinical use Purity. These conditions are unsatisfactory for routine clinical operation.

N ₂ S ₂ and N ₃ S systems as described by G. Bormans et al. (Nucl. Med. Biol. 1990, 17, 499-506) and described in European patent EP 0 173 424 and EP 0 250 013 meet the requirement for sufficient stability of the corresponding technetium-99m complexes, but are excreted from the organism too quickly and without specific enrichment, so that these find use only in the clinic as kidney function diagnostics and are therefore of limited use, in particular because the demand for substances that specifically accumulate in diseased tissues has increased.

There is an urgent need for stable bifunctional complexing agents to solve the refined nuclear medical questions, which do not have the disadvantages mentioned so far for complexation and which are capable of forming conjugates with bio- and macromolecules without their specificity and selectivity being essential to influence. At the same time, the requirements for the use of these compounds in humans with regard to the absorbed radiation dose, stability and solubility of the compounds must be fulfilled.

The object of the invention is therefore to make these compounds and conjugates available, to create the simplest possible process for their preparation and to provide a kit formulation of these compounds / conjugates according to the invention for their clinical use. This object is achieved by the present invention.

The compounds of the invention are represented by the general formula (I)

characterized in which

(A ¹ ), (A ² ) and (A ^* - ^* ) are the same or different and are for the general formula

are in which the two free valences are connected in any way with the respective nitrogen or sulfur atoms,

and in what

R ^* - ^** and R ^{5 are} identical or different, denote a hydrogen atom or a methyl or an ethyl group and

R ^{4 represents} a hydrogen atom, a branched or unbranched alkyl group having 1-4 carbon atoms, the C atoms of which may optionally contain amino groups, N (R ^a R ^t) ) groups (where R ^a and R ^* ° are the same or different - which are and represent branched or unbranched alkyl or acyl radicals having 1-20 carbon atoms, the C atoms of which are optionally substituted by a hydroxyl, a carboxy or an amino group), hydroxyl groups, thiol groups, halogens, carboxy groups, alkoxycarbonyl groups 1-20 carbon atoms, acyloxy groups with 1-12 carbon atoms, aminocarbonyl groups, sulfonyl groups, aminosulfonyl groups or phosphoric acid residues are substituted,

R ² and R ^{9 are the} same or different and have the same meaning as R ⁴ ,

k, 1 and m are the same or different and the numbers mean 0, 1, 2, 3 or 4 and

X is a hydrogen atom, a carboxy group, an alkoxy group with 1-20 carbon atoms, an alkoxycarbonyl group with 1-20 carbon atoms, an acyloxy group with 1-20 carbon atoms, an aminocarbonyl group, a sulfonyl group, an amino sulfonyl group, a phosphoric acid residue, a carboxymethylaminocarbonyl - Group, a p-aminophenyl group, a p-hydroxyphenyl group, a halogen atom, a hydroxy group, an amino group, an N (R ^a R ^* °) group (where R ^a and R ^** - ^{5 are the} same or different are and represent branched or unbranched alkyl or acyl radicals having 1-20 carbon atoms, the C atoms of which are optionally substituted by a hydroxyl, a carboxy or an amino group), a hydrazine group, a hydrazide group, one Cholesteryloxycarbonylmethylaminocarbonyl group, a cholesteryloxycarbonyl group, a cholesteryloxycarbonylmethyloxycarbonyl group, another steroid or a derivative of an ethynyl or ethenyl steroid, a substituent of the formula

in which

Q is a -NH- or -0-,

Z ^{1 has} the same meaning as Z,

Y ^{1 has} the same meaning as Y and i have the same meaning as m,

a substituent of the formula

V U

in which .

Q ^{1 is} a -NH-, -C0- or -0-,

U is a bond, a group of the formula - (OC ^ CO) ^ - with h = l-3 or a suitable linker for coupling to bio or macromolecules and V is a hydrogen atom, a hydroxy group, a

NfR ^ ^** - ^{5 "} ) group (where R ^a and R ^* ° are the same or different and stand for branched or unbranched alkyl or acyl radicals having 1-20 carbon atoms, the C atoms of which may be with a hydroxy, a carboxy - or an amino group), is a bio or macromolecule, means and

Y is an unsaturated, at least one double and / or triple bond-containing, unbranched or branched chain with up to 12 carbon atoms, which optionally, one or more times and at any point in the chain with hydroxyl, carboxy, alkoxy, amino or substituted amido groups with 1-20 carbon atoms in the alkyl and / or aryl radical means, Z is a hydrogen atom, a halogen atom, a carboxy group, a hydroxy group, an alkoxycarbonyl group with 1-20 carbon atoms, an acyloxy group with 1-20 carbon atoms, an alkoxy group with 1-20 carbon atoms, a cholesteryloxycarbonylmethylaminocarbonyl group, a cholesteryloxycarbonyl group, one Cholesteryloxycarbonylmethyloxycarbonyl group, another steroid or a derivative of an ethine or ethene steroid, a substituent of the formula

VU - Q ^J

in which

Q ^{1 is} a -NH-, -CO- or -0-,

U is a bond, a group of the formula - (OCH ₂ C = 0) _h - with h = l-3 or a suitable linker for coupling to bio or macromolecules and V is a hydrogen atom, a hydroxy group, a

N (R ^a R ^* °) group (where R ^a and R ^* ° are the same or different and represent branched or unbranched alkyl or acyl radicals having 1-20 carbon atoms, the C atoms of which may be hydroxyl or carboxy - or an amino group), is a bio or macromolecule, means

M represents an element of atomic number 43 or 75,

R ¹ the. has the same meaning as R ⁴ ,

R ⁶ , R ⁷ and R ^{8 are the} same or different and are for a hydrogen atom, an alkyl group with 1-4 carbon atoms, the C atoms of which are optionally substituted with hydroxyl, carboxy or amino groups or for are a group of the formula -CH ₂ -X, in which X has the meaning given above

and their water-soluble salts.

Preferred compounds of the general formula (I) according to the invention are characterized in that the radical Z is a hydrogen atom, a steroid, an ethynyl steroid or an ethenyl steroid.

Also preferred are compounds of the general formula (I) according to the invention, characterized in that the radical Z is a hydrogen atom, a steroid, an ethynyl steroid or an ethenyl steroid and the radical X is a hydrogen atom, a halogen atom, a carboxy group, an amino group or a Amido group with 1-20 carbon atoms in the alkyl and / or aryl radical means.

Also preferred are compounds of the general formula (I) according to the invention which are characterized in that the radical Z is a hydrogen atom, a steroid, an ethynyl steroid or an ethenyl steroid, the radical X is a hydrogen atom, a halogen atom, a carboxy group, an amino group or an amido group with 1-20 carbon atoms in the alkyl and / or aryl radical, Y is an ethinylidene group and the radicals R ¹ , R ³ , R ⁶ , R ⁷ and R ⁸ each represent hydrogen atoms and k, 1 and m each represent the number 0 .

Another group of preferred compounds of the general formula (I) according to the invention is characterized in that the radical Z is a hydrogen atom and the radical X is a cholesteryloxycarbonylmethylaminocarbonyl group, a cholesteryloxycarbonyl group, a Cholesteryloxycarbonylmethyloxycarbonylgruppe, another steroid or a derivative of an ethine or ethene steroid means.

Furthermore, preferred compounds of the general formula (I) according to the invention are characterized in that the radical Z is a hydrogen atom, the radical X is a cholesteryloxycarbonylmethylaminocarbonyl group, a cholesteryloxycarbonyl group, a cholesteryloxycarbonylmethyloxycarbonyl group, another steroid or a derivative of an ethyne or ethene steroid which means The radical Y represents an ethinylidene group and the radicals R ¹ , R ³ , R ⁶ , R ⁷ and R ⁸ each represent hydrogen atoms and k, 1 and m each represent the number 0.

A third group of preferred compounds of the general formula (I) according to the invention is characterized in that the radical Z represents a hydrogen atom and the radical X represents a hydrogen atom, a carboxy group or a substituent of the formula

V - U - Q ¹ -

is where

Q ^{1 is} a -NH-, -CO- or -O-,

U is a bond, a group of the formula - (0CH ₂ C0) _h - with h = l-3 or a suitable linker for coupling to bio or macromolecules and

V represents a hydrogen atom, a hydroxyl group, an N ^ '-'- R- ^** -') group (where R and R ^* ° are identical or different and represent branched or unbranched alkyl or acyl radicals having 1-20 carbon atoms, whose C atoms are optionally with a hydroxy, a carboxy or an amino group), is a bio or macromolecule.

Further preferred compounds of the general formula (I) according to the invention are characterized in that the radical Z represents a hydrogen atom and the radical X represents a hydrogen atom, a carboxy group or a substituent of the formula

V - U - Q ¹ -

is where

Q ^{1 is} a -NH-, -C0- or -0-, U is a bond, a group of the formula - (OC ^ CO) * ^ - with h = l-3 or a suitable linker for coupling to bio or macromolecules and

V is a hydrogen atom, a hydroxyl group, an N (R ^a R ^* °) group (where R ^a and R- ^{b are the} same or different and represent branched or unbranched alkyl or acyl radicals having 1-20 carbon atoms, the carbon atoms of which optionally substituted with a hydroxyl, a carboxy or an amino group), is a bio or macromolecule, the radical Y is an ethinylidene group and the radicals R ¹ , R ³ , R ⁶ , R ⁷ and R ⁸ each represent hydrogen atoms and k, 1 and each represent the number 0.

A fourth group of preferred ones according to the invention

Compounds of the general formula (I) are characterized in that the radical Z represents a hydrogen atom and the radical X represents a hydrogen atom, a carboxy group, an alkoxy group with 1-20 carbon atoms, an alkoxycarbonyl group with 1-20 carbon atoms Substance atoms, an acyloxy group with 1-20 carbon atoms, an aminocarbonyl group, a sulfonyl group, an aminosulfonyl group, a phosphoric acid residue, a carboxymethylaminocarbonyl group, a p-aminophenyl group, a p-hydroxyphenyl group, a halogen atom, a hydroxy group, an amino group, an N (R ^a R ^* °) - group (where R ^a and R ^* ° are the same or different and stand for branched or unbranched alkyl or acyl radicals having 1-20 carbon atoms, the C atoms of which may have a hydroxyl, a Carboxy or an amino group are substituted), a hydrazine group or a hydrazide group.

Furthermore, compounds of the general formula (I) according to the invention are characterized in that the radical Z denotes a hydrogen atom, the radical X denotes a hydrogen atom, a carboxy group, an alkoxy group with 1-20 carbon atoms, an alkoxycarbonyl group with 1-20 carbon atoms, an acyloxy group with 1-20 carbon atoms, an aminocarbonyl group, a sulfonyl group, an aminosulfonyl group, a phosphoric acid residue, a carboxymethylaminocarbonyl group, a p-aminophenyl group, a p-hydroxyphenyl group, a halogen atom, a hydroxy group, an amino group, an N (R ^a R ^* °) group (where R ^a and R ^* ° are identical or different and stand for branched or unbranched alkyl or acyl radicals having 1-20 carbon atoms, the C atoms of which may be substituted with a hydroxyl, a carboxy or or an amino group), a hydrazine group or a hydrazide group, the radical Y represents an ethinylidene group and the radicals R ¹ , R ³ , R ⁶ , R ⁷ and R ⁸ each represent hydrogen atoms and k, 1 and m each represent the number 0. The present invention further relates to compounds of the general formula (II)

in which A ¹ , A ² , A ³ , R ¹ , R ⁶ , R ⁷ , R ⁸ , Y and Z have the meaning given above and in which T is a hydrogen atom, an acetate group, a benzoate group, a p-methoxybenzyl group, a Represents acetamidomethyl group, a benzamidomethyl group, a trimethylacetamidomethyl group, a hydroxyacetyl group or another suitable sulfur protecting group.

The present invention furthermore relates to conjugates of compounds according to the invention of the general formulas I and II with substances which selectively accumulate in tissues. The fragments are linked depending on the type of substances to be connected in an amidic, imidic or ester-like manner. If the substances accumulating in the tissues are peptides, proteins, antibodies or fragments of these, the linkage is amidic via their amino groups; if the substances accumulating in tissues contain hydroxyl groups, the linkage is ester-like; if the substances accumulating in the tissues contain aldehyde groups, the linkage is imidic. Preference is given to substances which accumulate in tissues and which accumulate in diseased tissues, in particular in those tissues in which atherosclerotic plaques are present.

Also preferred are conjugates which are characterized in that the substances accumulating in the diseased tissue mean peptides and proteins, in particular endotheline, partial sequences of endothelin, endothelin analogues, endothelin derivatives or endothelin antagonists.

Particularly preferred conjugates are characterized in that the peptides contain the sequences or parts thereof

Cys-Ser-Cys-Ser-Ser-Leu-Met-Asp-Lys-Glu-Cys-Val-Tyr- Phe-Cys-His-Leu-Asp-Ile-Ile-Trp,

Cys-Ser-Cys-Ser-Ser-Trp-Leu-Asp-Lys-Glu-Cys-Val-Tyr-

Phe-Cy's-His-Leu-Asp-Ile-Ile-Trp,

Cys-Thr-Cys-Phe-Thr-Tyr-Lys-Asp-Lys-Glu-Cys-Val-Tyr-Tyr-Cys-His-Leu-Asp-Ile-Ile-Trp,

Cys-Ser-Ala-Ser-Ser-Leu-Met-Asp-Lys-Glu-Ala-Val-Tyr-

Phe-Cy7s-His-Leu-Asp-Ile-Ile-Trp,

Cys-Ser-Cys-Asn-Ser-Trp-Leu-Asp-Lys-Glu-Cys-Val-Tyr- Phe-Cys-His-Leu-Asp-Ile-Ile-Trp,

Cys-Ser-Cys-Lys-Asp-Met-Thr-Asp-Lys-Glu-Cys-Leu-Asn-Phe-Cys-His-Gln-Asp-Val-Ile-Trp, Ala-Ser-Cys-Ser-Ser-Leu-Met-Asp-Lys-Glu-Cys-Val-Tyr- Phe-Ala-His- Leu-Asp- Ile-Ile-Trp,

Ala-Ser-Ala-Ser-Ser-Leu-Met-Asp-Lys-Glu-Ala-Val-Tyr- Phe-Ala-His-Leu-Asp-Ile-Ile-Trp,

Cys-Ser-Cys-Ser-Ser-Trp-Leu-Asp-Lys-Glu-Ala-Val-Tyr- Phe-Ala-His-Leu-Asp-Ile-Ile-Trp, i 1 Cys-Val-Tyr- Phe-Cys-His-Leu-Asp-Ile-Ile-Trp,

N-Acetyl-Leu-Met-Asp-Lys-Glu-Ala-Val-Tyr-Phe-Ala-His-Leu-Asp-Ile-Ile-Trp

the partial sequences

His-Leu-Asp-Ile-Ile-Trp,

or the cyclic amino acid sequences

Cyclo- (DTrp-DAsp-Pro-DVal-Leu),

Cyclo- (DGlu-Ala-alloDIle-Leu-DTrp)

exhibit.

The preparation of the compounds of the invention

takes place according to the processes known to the person skilled in the art, for example by reaction of technetium-99m in the form of pertechnetate, which can be eluted from molybdenum-technetium generators with little effort, in the presence of a reducing agent (for example Sn (II) chloride or dithionide) and optionally an auxiliary ligand (for example sodium citrate or sodium tartrate) with a compound of the general formula (II)

wherein

have the meaning given.

The reaction is preferably carried out in an aqueous medium at room temperature. The SH-

Protecting group takes place in situ or according to the literature methods known to the person skilled in the art, for example by basic hydrolysis, reductive cleavage, etc. (see for example "Protective Groups in Organic Synthesis", T.W. Greene, John Wiley and Sons 1981).

The present invention furthermore relates to processes for the preparation of the compounds of the general formula (II) according to the invention

characterized in that one is known per se

Way a) a compound of general formula (III)

wherein

A ¹ , A ² , A ³ , R ¹ , R ⁶ , R ⁷ , R ⁸ , Y and Z the above

Have meaning and shark is a halogen, with a compound of the general formula

T - S ^" M ⁺

in which M ⁺ denotes an alkali metal cation and T has the meaning given above,

or b) a compound of the general formula

HN (R ⁶ ) -A ² -N (R ⁷ ) -A ³ -N (R ⁸ ) -YZ

wherein A ² , A ³ , R ⁶ , R ⁷ and R ^{8 have} the meaning given above and with a compound of the general formula

W

wherein A ¹ and T have the meaning given in claim 11 and W has the meaning of a leaving group which enables A ¹ to react with free amino groups,

implements.

The N-haloacetylamides required as starting substances are prepared by methods known from the literature by acylation of an amino function with haloacetic acid halides (JACS, 1969, 90, 4508; Chem. Pharm. Bull. 1981, 29 (1), 128).

The required amines of the general formula

HN (R ⁶ ) -A ² -N (R ⁷ ) -A ³ -N (R ⁸ ) -YZ

are synthesized according to methods of peptide synthesis known from the literature (see for example in "The Practice of Peptide Synthesis", M. Bodanszky and A. Bodanszky; Springer-Ver¬ lag, 1984 and Fieser, Reagents for Organic Synthesis 10, 142). N-protected .alpha.- and .beta.-amino acids are processed according to the methods known to those skilled in the art, for example via an addition / elimination reaction of an amine (primary or secondary) with a Carbonyl compound (for example acid chloride, mixed anhydride, activated ester) coupled to N-unprotected organic molecules. After the amino protecting group has been cleaved off using known literature methods, for example by acidic or basic hydrolysis, hydrogenolysis, reductive cleavage with alkali metals in liquid ammonia, the remaining amino function is again reacted with an N-protected a- and ß-amino acid derivatized according to known literature methods (for example carbodiimide method). The amino protective group is split off according to the literature methods mentioned above.

The further required compounds of the general formula

W

are converted by the methods known to the person skilled in the art by converting the carboxylic acid (s) corresponding carboxylic acids, for example by the carbodiimide method (Fieser, Reagents for Organic Synthesis 10, 142), over a mixed or cyclic anhydride (Org. Prep. Proc Int. 1975, 7, 215) or via an activated ester (Adv. Org. Chem. Part B, 472).

The compounds according to the invention have the desired properties to a high degree. They contain the radioisotopes required for their use in a stable bound form in the complex.

The coupling possibilities of the chelate complexing agents according to the invention are of particular importance since they are to be regarded as trifunctional and, in addition to metal complexation, enable coupling via the multiple bond as well as via a carboxy or amino group in A ¹ , A ² or A ³ .

Of particular interest according to the invention is the provision of labeling methods which, under milder conditions than previously known methods, enable the compounds according to the invention to be implemented rapidly and quantitatively immediately before the use of the radio pharmacy.

Linking the chelating agents via the multiple bond with steroids is of great advantage, it being possible either to couple to a -CH ₂ group of the steroid or to link to the 17-ethynyl or 17-ethenyl group of steroids.

The affinity of some technetium-99m labeled steroid derivatives to their respective is surprisingly good

Steroid hormone receptor. The technetium complex of 3,17ß-dihydroxy-17o; - (5- [2-benzoylthioacetylglycyl-glycyl] -aminopent-1- (E) -en-3-in) -1,3,5-estratriene ( Example 13b) in the in vitro cytosol test (Carlson, KE et al. 1989, 32, 345-355) a receptor affinity which corresponds to that of 17ß-estradiol. As a result, these compounds are outstandingly suitable for receptor imaging with technetium-99m and for tumor diagnosis / therapy of steroid hormone-dependent tumors.

Another advantage of the present invention is that the optional linkage via the carboxy or multiple bond offers the possibility of solubility and pharmacokinetics of these complexes by chemical substitution at the grain level. control plexer. Biodegradable groups or linkers in X are of particular importance here.

Compounds with lipophilic substituents in X or Z show surprising properties, e.g. in the case of the compound of Example 9, which accumulate in atherosclerotic plaques and thus show a possibility of representing atherosclerotic changes on the walls of the vessels.

By selecting suitable bio- and macromolecules (eg monoclonal antibodies, steroids, ...) in X or Z, complexes according to the invention are obtained which have a surprisingly high tissue and organ specificity and are therefore excellent for solving a number of diagnostic and therapeutic Problems can contribute.

The complex-forming ligands of the general formula (II) in which Z denotes a hydrogen atom and / or ligands of the general formula (II) in which an ethynyl function is contained can be obtained by the processes known to the person skilled in the art (for example R. Rossi et al., Tetrahedron 1983, 39, 287) can be coupled to iodine vinyl steroids. The iodine vinyl steroids required are synthesized using methods known from the literature (for example H. Hofmeister et al., Tetrahedron 1986, 42, 3575).

The conversion of carboxy groups on compounds of the general formula (II) can be carried out, for example, by the carbodiimide method (Fieser, Reagents for Organic Synthesis 10, 142), via a mixed or cyclic anhydride (Org. Prep. Proc. Int. 1975, 7 , 215) or above an activated ester (Adv. Org. Chem. Part B, 472) can be carried out.

The coupling of the chelating agents to bio or macromolecules takes place according to the methods known to the person skilled in the art by converting the carboxy group (s) on the chelating agent, for example according to the carbodiimide method (Fieser, Reagents for Organic Synthesis 10, 142), via a mixed one or cyclic anhydride (Org. Prep. Proc. Int. 1975, 7, 215) or via an activated ester

(Adv. Org. Chem. Part B, 472) and subsequent reaction with a nucleophilic group of the bio or macro molecule to form a covalent bond.

The chelating agents obtained in this way can also be linked to bio or macromolecules which are known to accumulate particularly in the organ, organ part or tissue to be examined. Such molecules are, for example, enzymes, hormones, polysaccharides such as dextrans or starches, porphyrins, bleomycins, insulin, prostaglandins, steroid hormones, amino sugars, amino acids, peptides such as polylysine, proteins (such as immunoglobulins, monoclonal antibodies) , Lectins), lipids (also in the form of liposomes) and nucleotides of the DNA or RNA type. Particularly noteworthy are conjugates with albumins, such as human serum albumin, antibodies, such as monoclonal antibodies or antimyosin, specific for tumor-associated antigens. Instead of biological macromolecules, suitable synthetic polymers such as polyethyleneimines, polyamides, ect. be tied up.

The pharmaceutical agents formed therefrom are suitable, for example, for use in tumor diagnosis and tumor therapy, atherosclerosis diagnosis. stik or receptor imaging. The binding affinity and binding specificity of the pharmaceutical agents formed to the target organ, target organ part or target tissue must not be affected or only slightly impaired by the conjugate formation.

The radiopharmaceutical compositions according to the invention are likewise prepared in a manner known per se by dissolving or suspending the complexing agents according to the invention and their conjugates - optionally with the addition of the additives customary in galenicals - in an aqueous medium and then optionally lyophilizing the solution or suspension . Suitable additives are, for example, physiologically acceptable buffers (such as tromethamine), additives of auxiliary ligands (such as sodium citrate or sodium tartrate), reducing agents (such as tin (II) chloride) or - if necessary - electrolytes such as sodium chloride or - if necessary - (an) auxiliary (s) customary in galenics (for example lactose, mannitol) and / or surfactant (s) (for example lecithins, Tween ^ ^R ^, Myrj ^(R ). The additives used must allow the preparation of the compounds according to the invention in their composition.

The invention further relates to a kit for the production of radiopharmaceuticals, consisting of a compound of the general formula (II), a reducing agent and, if appropriate, an auxiliary ligand, which are present in the dry state or in solution, an instruction for use with a reaction instruction for implementing the described Compounds with Techne¬ tium-99m or rhenium in the form of a pertechnetate solution or perrhenate solution. The radiopharmaceutical compositions according to the invention are administered to a patient in an amount of 0.1 mCi to 50 mCi, preferably in an amount of 0.5 mCi to 30 mCi per 70 kg of body weight. Details of their use and dosage are described, for example, in "Radiotracers for Medical Applications", CRC-Press, Boca Raton, Florida. They are intended for intravenous application. The complex compounds / conjugates according to the invention are used for radio diagnostics and radiotherapy in the form of their complexes with the radioisotopes of the elements with the atomic number 43 or 75.

The radiopharmaceutical compositions according to the invention meet the diverse requirements for suitability as radiopharmaceuticals for radio diagnostics and radiotherapy. So they are ideally suited to Enrich application in target tissues and thus enable a non-invasive

Diagnosing appropriate tissues. If necessary, the water-solubility of the radiopharmaceutical compositions according to the invention is ensured by the auxiliaries customary in galenics, as described above. Furthermore, the radiopharmaceutical agents according to the invention not only have a high stability in vitro, but also a surprisingly high stability in vivo, so that the radionuclide bound in the complex is not released or is not clinically relevant or is not exchanged.

Preferred radiopharmaceutical compositions according to the invention are further characterized in that they contain the compounds of the general formula I or II according to the invention in the form of liposomes and that if appropriate, the compound of the general formula (I) according to the invention is prepared in a kit with technetium or rhenium in the form of the permetallates.

The radiopharmaceutical agents according to the invention can also be used in radioimmunotherapy or radiation therapy. These differ from radio diagnostics only in the amount and type of radioisotope used. The goal is the destruction of tumor cells by high-energy short-wave radiation with the shortest possible range. Suitable ß-emitting isotopes are, for example, rhenium-186 and rhenium-188.

The radiopharmaceutical compositions according to the invention are suitable for the non-invasive in vivo presentation of tissues containing steroid receptors, for example steroid hormone-dependent tumors.

The radiopharmaceutical agents according to the invention are suitable for the non-invasive in vivo imaging of atherosclerotic vessel changes, for example of plaques.

Overall, it has been possible to synthesize new complexing agents and metal complexes which open up new possibilities in diagnostic and therapeutic medicine. Above all, the development of novel imaging methods in nuclear medicine diagnostics makes this development appear desirable. The following examples illustrate the invention.

example 1

a) S-benzoylthioacetylglycylglycylpropargylamide [la]

A solution of 215.1 mg (1.22 mmol) of potassium thiobenzoate in anhydrous DMF is added dropwise under nitrogen to a solution of 150 mg (0.61 mmol) of chloroacetylglycylglycylpropargylamide in anhydrous DMF. Catalytic amounts of sodium iodide are then added and the mixture is heated to 110 ° C. for 2 hours. After the reaction mixture has cooled to room temperature, it is stirred into 1 N HCl and extracted with ethyl acetate until no more product is present in the water phase. The organic phases are evaporated to dryness under reduced pressure, taken up again in ethyl acetate, washed with water and dried over Mg ₂ SO ₄ . After the solvent has been stripped off under reduced pressure, the residue is recrystallized from methanol or, if necessary, purified by column chromatography with CH Cl ₂ / MeOH (9: 1).

Yield: 76% of the title compound [la]

^C 16 ^H 17 ^N 3 ° 4 ^{S (MW: 347 3} 9 ^{) 1} H-NMR (dg-DMSO): δ = 3.11 (t; 1H, -C = CH)

3.69 (d; 2H, -NH (CH ₂ ) CO-)

3.78 (d; 2H, -NH (CH ₂ ) CO-)

3.84 - 3.88 (m; 2H, -NH (CH ₂ ) -C≡CH) 3.90 (S; 2H, -SCH ₂ CO-)

7.55 - 7.61 (m; 2H, ArH)

7.69 - 7.74 (m; 1H, ArH)

7.93 - 7.97 (m; 2H, ArH)

8.20 (t; 1H, -NH-) 8, 27 (t; 1H, -NH-) 8, 50 (t; 1H, -NH-)

b) Technetium-99m complex [lb] of S-benzoylthioacetylglycylglycylpropargyla id

A solution of 0.5 mg (1.44 μmol) S-benzoylthioacetylglycylglycylpropargylamide [la] in 50 μl 1 N sodium hydroxide solution is mixed with 250 μl phosphate buffer (Na ₂ HP0 ₄ , 0.5 mol / 1, pH 8.5) diluted. Then 50 μl of a 0.15 molar trisodium citrate dihydrate solution and 2.5 μl of a 0.2 molar tin (II) chloride dihydrate solution are added. The reaction mixture is mixed with a pertechnetate solution (0.4-0.9 mCi) from a Mo-99 / Tc-99m generator, incubated for 10 minutes at room temperature and then filtered (0.2 μm filter). The analysis of the marking is carried out by means of HPLC: MERCK Nucleosil column, 125 x 4 mm, 5 μm; Gradient from 100% A to 100% B within 7.5 min; Eluent A: phosphate buffer (Na ₂ HP0; 0.01 M; pH 2.0); Eluent B: acetonitrile / phosphate buffer (Na HP0 ₄ ; 0.01 mol; pH 2.0) 50:50 (v / v); Flow rate: 1.0 ml / min. The radiochemical purity of the Tc-99m complex is more than 95%.

Example 2

a) 2-acetylthiosuccinylglycylglycylpropargylamide [2a]

5.8 g (37.5 mmol) of glycylglycylpropargylamide are dissolved in 60 ml of anhydrous DMF under nitrogen, with 8.50 g (48.7 mmol) of acetylmercapto-succinic anhydride added and stirred at room temperature overnight. The reaction mixture is then evaporated to dryness under reduced pressure. The residue is suspended in methanol, suction filtered and recrystallized from water.

Yield: 61% of the title compound [2a]

C ₁₃ H ₁₇ N ₃ 0 ₆ S (MW: 343.36) - ^• H NMR (d ₆ -DMSO): δ = 2.35 (S; 3H, -COCH3) 2.66; 2.83 (2x dd; 2H, - (CH ₂ ) COOH) 2.99 (t; 1H, -C≡CH)

3.70 - 3.77 (m; 4H, 2x -NH (CH ₂ ) C0-) 3.88 (dd; 2H, -NH (CH ₂ ) -C≡CH) 4.35 (t; 1H, - S-CH (CH ₂ COOH) -CH ₂ ~) 8.07 (t; 1H, -NH-) 8.12 (t; 1H, -NH-) 8.26 (t; 1H, -NH-)

b) Technetium-99m complex [2b] of 2-acetylthiosuccinylglycylglycylpropargylamide

A solution of 0.5 mg (1.45 μmol) 2-acetylthiosuccinylglycylglycylpropargylamid [2a] in 50 μl 0.1 N sodium hydroxide solution is mixed with 250 μl phosphate buffer (Na ₂ HP0 ₄ ,

0.5 mol / 1, pH 8.5). Then 50 μl of a 0.15-molar trisodium citrate dihydrate solution and 2.5 μl of a 0.2-molar tin (II) chloride dihydrate solution are added. A pertechnetate solution (0.4-0.9 mCi) from a Mo-99 / Tc-99m generator is added to the reaction mixture, incubated for 10 minutes at room temperature and then filtered (0.2 μm filter) . The analysis of the marking is carried out by means of HPLC: MERCK Nucleosil column, 125 x 4 mm, 5 μm; Gradient from 100% A to 100% B within 7.5 min; Eluent A: phosphate buffer (Na ₂ HP0 ₄ ; 0.01 M; pH 2.0); Eluent B: acetonitrile / phosphate buffer (Na ₂ HP0 ₄ ; 0.01 M; pH 2.0) 50:50 (v / v); Flow rate: 1.0 ml / min. The radiochemical purity of the Tc-99m complex is more than 95%.

Example 3

a) S-acetyl-hioacetylglutaminylglycylpropargylamide [3a]

A solution / suspension of 2.0 g (8.3 mmol) of glutaminyl-glycylpropargylamide in 80 ml of anhydrous DMF is slowly mixed with a solution of 5.3 g (8.4 mmol) of S-acetyl-mercaptoacetic acid-N-hydroxysuccinimide ester in water ¬ free THF added. After 4 hours of stirring at room temperature, the mixture is hydrolyzed with water. The solvent is then removed under reduced pressure and the residue is recrystallized from methanol / water. Yield: 45% of the title compound [3a]

C ₁₄ H ₁₉ N ₃ 0gS (MW: 357.36) - ^• -H NMR (dg-DMSO): δ = 1.64-1.92 (m; 2H, -CHCH ₂ CH ₂ COOH) 2.31 (S; 3H, -COCH3) 2.36 (t; 2H, -CHCH ₂ CH ₂ COOH) 3.02 (t; 1H, -C≡CH) 3.70 (s; 2H, -NH (CH ₂ ) CO-) 3.73 - 3.80 (m; 1H, -CHCH ₂ CH ₂ COOH) 3.89 (dd; 2H, -NH (CH ₂ ) -C≡CH) 4.21 (s; 2H, - SCH ₂ CO-) 8.09 (m; 1H, -NH-) 8.19 (m; 1H, -NH-) 8.26 (m; 1H, -NH-) b) Technetium-99m complex [3b] of S-acetyl-thioacetylglutaminylglycylpropargyla id

A solution of 0.5 mg (1.40 μmol) of S-acetyl-thioacetyl-glutaminylglycylpropargylamide [3b] in 50 μl of 0.1 N sodium hydroxide solution is mixed with 250 μl of phosphate buffer (Na ₂ HP0 ₄ , 0.5 mol / 1, pH 8.5) diluted. Then 50 μl of a 0.15-molar trisodium citrate dihydrate solution and 2.5 μl of a 0.2-molar tin (II) chloride dihydrate solution are added. A pertechnetate solution (0.4-0.9 mCi) from a Mo-99 / Tc-99m generator is added to the reaction mixture, incubated for 10 minutes at room temperature and then filtered (0.2 μm filter) .

The analysis of the marking is carried out by means of HPLC: MERCK Nucleosil column, 125 x 4 mm, 5 μm; Gradient from 100% A to 100% B within 7.5 min, eluent A: phosphate buffer (Na ₂ HP0 ₄ ; 0.01 M; pH 2.0); Eluent B: acetonitrile / phosphate buffer (Na ₂ HP0 ₄ ; 0.01 M; pH 2.0) 50:50 (v / v); Flow rate: 1.0 ml / min. The radiochemical purity of the Tc-99m complex is more than 95%.

Example 4

a) S-acetyl-thioacetylglycylglutaminylpropargylamide [4a]

A solution / suspension of 2.0 g (8.3 mmol) of glycylglutaminylpropargylamide in 80 ml of anhydrous DMF is slowly mixed with a solution of 5.3 g (8.4 mmol) of S-acetyl-mercaptoacetic acid-N-hydroxysuccinimide ester in water - free THF. After 4 hours of stirring at room temperature, the mixture is hydrolyzed with water. The solvent is then removed under reduced pressure and the residue is recrystallized from methanol / water. Yield: 61% of the title compound [4a] C ₁₄ H ₁₉ N ₃ 0 ₆ S (MW: 357.36) ¹ H-NMR (d ₆ -DMSO):

Ö = 1.60-1.88 (m; 2H, -CHCH ₂ CH ₂ COOH) 2.30 (S; 3H, -C0CH ₃ ) 2.28 (t; 2H, -CHCH ₂ CH ₂ COOH) 3, 04 (t; 1H, -C≡CH) 3.68 (S; 2H, -NH (CH ₂ ) C0-) 3.70 - 3.77 (m; 1H, -CHCH ₂ CH ₂ COOH) 3.88 (dd; 2H, -NH (CH ₂ ) -C≡CH) 4.19 (s; 2H, -SCH ₂ CO-) 8.11 (m; 1H, -NH-) 8.23 (m; 1H, -NH-) 8.35 (m; 1H, -NH-)

b) Technetium 9 m complex [4b] of S-acetyl-thioacetylglycylglutaminylpropargylamide

A solution of 0.5 mg (1.40 μmol) of S-acetylthioacetylglycylglutaminylpropargylamide [4a] in 50 μl of 0.1 N sodium hydroxide solution is mixed with 250 μl of phosphate buffer (Na ₂ HP0 ₄ ,

0.5 mol / 1, pH 7.5). Are then added 50 .mu.l of a 0.15 molar trisodium citrate solution, and 2, ^'5 of a 0.2 molar tin (II) chloride dihydrate ul solution. A pertechnetate solution (0.4-0.9 mCi) from a Mo-99 / Tc-99m generator is added to the reaction mixture, incubated for 10 minutes at room temperature and then filtered (0.2 μm filter) . The analysis of the marking is carried out by means of HPLC: MERCK Nucleosil column, 125 x 4 mm, 5 μm; Gradient from 100% A to 100% B within 7.5 min; Eluent A: phosphate buffer (Na ₂ HP0 ₄ ; 0.01 M; pH 2.0); Eluent B: acetonitrile / phosphate buffer (Na ₂ HP0 ₄ ; 0.01 M; pH 2.0) 50:50 (v / v); Flow rate: 1.0 ml / min. The radiochemical purity of the Tc-99m complex is more than 95%.

Example 5

a) S- (p-methoxybenzyl) cysteinylglycylglycylpropargylamide [5a]

9.8 g (0.02 mol) of N-tert-butoxycarbonyl-S- (p-methoxy-benzyl) cysteinylglycylglycylpropargylamide are dissolved in trifluoroacetic acid at 0 ° C. and stirred in an ice bath for 4 hours. The mixture is then evaporated to dryness under reduced pressure, ethanol is added to the residue and the mixture is concentrated again. The residue is purified by column chromatography on silica gel [CH ₂ C1 ₂ / CH ₃ 0H

(5: 1)] cleaned. The product crystallizes after concentration.

Yield: 93% of the title compound [5a] C ₁₈ H ₂₄ N ₄ 0 ₄ S (MW: 392.48) ¹ H-NMR (CD ₃ OD): δ = 2.54 (t; 1H, -C≡CH)

2.63 - 2.86 (m; 2H, -CHCH ₂ S-) 3.48 - 3.52 (m; 1H, -CHCH ₂ S-) 3.71 (s; 2H, -NH (CH ₂ ) C0-) 3.77 (s; 3H, -0CH ₃ )

3.85 (S; 2H, -NH (CH ₂ ) CO-) 3.92 (S; 2H, -CH ₂ Ar) 3.97 (dd; 2H, -NH (CH ₂ ) -C≡CH) 6 , 86; 7.25 (AA'BB '; 4H, ArH) b) Technetium-99m complex [5b] of cysteinylglycylglycylpropargylamide

A solution of 0.5 mg (1.27 μmol) of S- (p-methoxybenzyl) cysteinylglycylglycylpropargylamide [5a] in liquid HF is stirred at 0 ° C for 15 minutes. After this

Evaporation of the solvent at room temperature, the residue is taken up in 50 μl of 1 N sodium hydroxide solution and diluted with 250 μl of phosphate buffer (Na ₂ HP0 ₄ , 0.5 mol / 1, pH 7.5). Then 50 μl of a 0.15 molar trisodium citrate dihydrate solution and 2.5 μl of a 0.2 molar tin (II) chloride dihydrate solution are added.

The reaction mixture is with a pertechnetate solution

(0.4-0.9 mCi) from a Mo-99 / Tc-99m generator, incubated for 10 minutes at room temperature and then filtered (0.2 μm filter).

The analysis of the marking is done by HPLC:

MERCK Nucleosil column, 125 x 4 mm, 5 µm;

Gradient from 100% A to 100% B within 7.5 min; Eluent A: phosphate buffer (Na ₂ HP0 ₄ ; 0.01 M; pH 2.0);

Eluent B: acetonitrile / phosphate buffer (Na ₂ HP0 ₄ ; 0.01 M; pH 2.0) 50:50 (v / v); Flow rate: 1.0 ml / min.

The radiochemical purity of the Tc-99m complex is more than 95%.

Example 6

a) N- [2-acetylthio-3-cholesteryloxycarbonyl-propionyl] -glycylglycylpropargylamide [6a]

A solution of 5.0 g (8.9 mmol) of 3-acetylthiosuccinic acid cholesterol ester in anhydrous THF is cooled in an ice bath and an excess of dry triethylamine is added. Then 1.9 g (17.8 mmol) Ethyl chloroformate added dropwise. The mixture is allowed to warm to 0 ° C., mixed with 3.0 g (18.0 mmol) of glycylglycylpropargylamide and stirred for 3 hours at room temperature. The reaction mixture is hydrolyzed with water and extracted with ethyl acetate until no more product is present in the water phase. The organic phases are evaporated to dryness under reduced pressure, taken up again in ethyl acetate, washed with water and dried over Mg ₂ SO ₄ . After the solvent has been removed under reduced pressure, the residue is recrystallized from methanol or, if appropriate, purified by means of column chromatography with CH ₂ Cl ₂ / MeOH (9: 1). Yield: 67% of the title compound [6a] C ₄₀ H ₆₁ N ₃ O ₆ S (MW: 712.01)

- ^• -H-NMR (CDC1 ₃ ): d = 0.68 - 2.08 (m; 41H, steroidal)

2.30 (d; 2H, -C = CHCH ₂ - steroidal) 2.32 (s; 3H, -C0CH ₃ )

2.76 - 3.00 (m; 2H, - (CH ₂ ) COO-)

3.10 (t; 1H, -C≡CH)

3.66 - 3.73 (m; 4H, 2x -NH (CH ₂ ) C0-) 3.84 (d; 2H, -NH (CH ₂ ) -C≡CH) 4.39 (t; 1H, - S-CH (CH ₂ COOH) -CH ₂ -)

4.52 - 4.63 (m; 1H, - (CH) o-steroidal) 5.48 (d; 1H, -C = CH-steroidal) 8.05 (t; 1H, -NH-)

8.11 (t; 1H, -NH-) 8.23 (t; 1H, -NH-) b) Technetium-99m complex [6b] of the N- [2-acetylthio-3-cholesteryloxycarbonylpropionyl] glycylglycylprop pargylamide

A solution of 0.5 mg (0.7 μmol) of N- [2-acetylthio-3-cholesteryloxycarbonylpropionyl] glycylglycylpropagylamide [6a] in 300 μl of DMSO is mixed with 30 μl of 1 N sodium hydroxide solution. Then 50 μl of a 0.15 molar trisodium citrate dihydrate solution and 0.4-0.9 mCi of a pertechnetate solution from a Mo-99 / Tc-99m generator are added. The reaction mixture is added in portions with 10 ul of a 0.2 molar tin (II) chloride dihydrate solution, incubated for 10 minutes at room temperature and then filtered (0.2 micron filter). The analysis of the label is carried out using HPLC: MERCK Nucleosil column, 125 x 4 mm, 5 μm; Gradient from 100% A to 100% B within 30 min; Eluent A: acetonitrile / water 50:50 (v / v), eluent B: acetonitrile; Flow rate: 1.0 ml / min. The radiochemical purity of the Tc-99m complex is more than 95%.

Example 7

a) Choles erin [N- (2-acetylthiosuccinylglycylglycylpropargylamid-4-yl) glycine] ester [7a]

A solution of 2.0 g (5.8 mmol) of 2-acetylthiosuccinylglycylglycylpropargylamide [2a] in anhydrous THF is cooled in an ice bath and an excess of dry triethylamine is added. 1.3 g (11.6 mmol) of ethyl chloroformate are then added dropwise. The mixture is allowed to warm to 0 ° C., mixed with 5.0 g (12.0 mmol) of Gly- cincholesterol ester and stirred for 3 hours at room temperature. The reaction mixture is hydrolyzed with water and extracted with ethyl acetate until no more product is present in the water phase. The organic phases are washed with water and dried over Mg ₂ S0 ₄ . After the solvent has been removed under reduced pressure, the mixture is recrystallized from methanol or, if appropriate, purified by means of column chromatography with CH ₂ Cl ₂ / MeOH (9: 1). Yield: 41% of the title compound [7a] C ₄₂ H ₆₄ N ₃ 0 ₆ S (MW: 769.06) - ^• -H-NMR (CD ₃ 0D): δ = 0.68 - 2.10 (m; 41H steroidal)

2.32 (d; 2H, -C = CHCH ₂ - steroidal) 2.33 (s; 3H, -COCH ₃ )

2.81-3.10 (m; 2H, -CH (CH ₂ ) C00-) 3.08 (t; 1H, -C≡CH) 3.41 (d; 2H, -NH (CH ₂ ) C0 -) 3.64 - 3.71 (m; 4H, 2x -NH (CH ₂ ) CO-) 3.83 (d; 2H, -NH (CH2) -C≡CH)

4.35 (t; 1H, -S-CH (CH ₂ COOH) -CH ₂ -) 4.61 - 4.72 (m; 1H, - (CH) o-steroidal) 5.38 (d; 1H, -C = CH- steroidal) 8.05 (t; 1H, -NH-) 8.09 (t; 1H, -NH-) 8.13 (t; 1H, -NH-) 8.25 (t; 1H , -NH-)

b) Technetium-99m complex [7b] of the cholesterol [N- (2-acetylthiosuccinylglycylglycylpropargylamid-4-yl) glycine] ester

A solution of 0.5 mg (0.65 µmol) cholesterol [N- (2-acetylthiosuccinylglycylglycylpropargylamid-4-yl) -gly- an] ester [7a] in 300 μl DMSO is mixed with 30 μl 0.1 N sodium hydroxide solution. Then 50 μl of a 0.15 molar trisodium citrate dihydrate solution and 0.4-0.9 mCi of a pertechnetate solution from a Mo-99 / Tc-99m generator are added. 10 μl of a 0.2 molar tin (II) chloride dihydra solution are added to the reaction mixture in portions, incubated for 10 minutes at room temperature and then filtered (0.2 μm filter). The analysis of the marking is carried out by means of HPLC: gradient from 100% A to 100% B within 30 min; Eluent A: acetonitrile / water 50:50 (v / v); Eluent B: acetonitrile; Flow rate: 1.0 ml / min. The radiochemical purity of the Tc-99m complex is more than 95%.

Example 8

a) N-Dodecanoyl-S- (p-methoxybenzyl) cysteinylglycylglycylpropargylamide [8a]

2.0 g (5.1 mmol) of S- (p-methoxybenzyl) cysteinylglycylglycylpropargylamide [5a] are dissolved in CH ₂ C1 ₂ and a little NEt _{3 is} added. Then 1.2 g (5.5 mmol) of dodecanoic acid chloride in CH ₂ C1 _{2 are} added dropwise and the mixture is stirred at room temperature for 2 hours. The reaction mixture is hydrolyzed with water and extracted with CH ₂ C1 ₂ . After drying, the mixture is concentrated under reduced pressure and the residue is chromatographed on silica gel using CH Cl ₂ / MeOH (9: 1). Yield: 80% of the title compound [8a]

C ₃₀ H ₄₆ N ₄ O ₅ S (MW: 574.79) --- H-NMR (dg-DMSO): δ = 0.85 (t; 3H, - (CH ₂ ) _g CH ₃ ) 1.19 - 1.30 (m; 16H, - (CH ₂ ) _g CH ₃ )

1.46 - 1.54 (m; 2H, -CH ₂ -CH ₂ CONH-)

2.08 - 2.20 (m; 2H, -CH ₂ -CH ₂ CONH-)

2.51 - 2.79 (m; 2H, -CHCH ₂ S-) 3.08 (t; 1H, -C≡CH)

3.74 (s; 3H, -OCH ₃ )

3.67 - 3.78 (m; 4H, 2x -NH (CH ₂ ) CO-)

3.76 (d; 2H, -CH ₂ Ar)

3.85 - 3.88 (m; 2H, -NH (CH ₂ ) -C≡CH) 4.49 - 4.55 (m; 1H, -CHCH ₂ S-)

6.85; 7.23 (AA'BB ¹ ; 4H, ArH)

8.04 - 8.09 (m; 2H, 2x -NH-)

8.22 (t; 1H, -NH-)

8.33 (t; 1H, -NH-)

b) Technetium-99m complex [8b] of N-dodecanoyl-cysteinylglycylglycylpropargylamide

A solution of 0.5 mg (0.9 μmol) of N-dodecanoyl-cysteineylglyeylglycylpropargylamide [8a] in liquid HF is stirred for 15 min at 0 ° C. After the solvent has been evaporated off at room temperature, the residue is dissolved in 300 μl of DMSO and 30 μl of 1N sodium hydroxide solution are added. Subsequently, 50 μl of a 0.15 molar trisodium citrate dihydrate solution and 0.4-0.9 mCi of a pertechnetate solution from a Mo-99 / Tc-99m generator are added. The reaction mixture is mixed in portions with 10 μl of a 0.2 molar tin (II) chloride dihydrate solution, incubated for 10 minutes at room temperature and then filtered (0.2 μm filter).

The analysis of the marking is carried out by means of HPLC: MERCK Nucleosil column, 125 x 4 mm, 5 μm; Gradient from 100% A to 100% B within 7.5 min, eluent A: phosphate buffer (Na ₂ HP0; 0.01 M; pH 2.0); Eluent B: acetonitrile / phosphate buffer (Na ₂ HP0 ₄ ; 0.01 M; pH 2.0) 50:50 (v / v); Flow rate: 1.0 ml / min.

The radiochemical purity of the Tc-99m complex is more than 95%.

Example 9

a) N- (3-Hexadecylaminocarbonyl-2-acetylthio-propionyl) glycylglycylpropargylamide [9a]

A solution of 2.0 g (5.8 mmol) of 2-acetylthiosuccinylglycylglycylpropargylamide [2a] in anhydrous THF is cooled in an ice bath and mixed with 1.7 g (7.0 mmol) of hexadecylamine. 1.47 g (7.0 mmol) of dicyclohexylcarbodiimide in anhydrous THF to which 2 ml of triethylamine have been added are then added dropwise at 0.degree. The mixture is allowed to warm to room temperature and stirred at this temperature for 3 hours. A few drops of acetic acid are added to the reaction mixture, hydrolysed with water and extracted with ethyl acetate until no more product is present in the water phase. The organic phases are washed with water and dried over Mg ₂ S0 ₄ . After the solvent has been concentrated under reduced pressure, it is recrystallized from methanol and, if appropriate, purified by column chromatography with CH ₂ Cl ₂ / MeOH (9: 1). Yield: 69% of the title compound [9a] C ₂₉ H ₅₀ N ₄ O ₅ S (MW: 566.81) -NMR (dg-DMSO): δ = 0.87 (t; 3H, -CH ₂ CH ₃ )

1.16 - 1.35 (m; 28H, - (CH ₂ ) ₁₄ CH ₃ ) 2.33 (s; 3H, -C0CH ₃ ) 2.73 - 3.10 (m; 4H, - (CH ₂ ) -) 3.07 (t; 1H, -C≡CH)

3.67 - 3.76 (m; 4H, 2x -NH (CH ₂ ) C0-)

3.85 - 3.89 (m; 2H, -NH (CH ₂ ) -C≡CH)

4.29 - 4.35 (m; 1H, -S-CH (CH ₂ C00H) -CH ₂ -) 7.99 (t; 1H, -NH-)

8.07 (t; 1H, -NH-)

8.15 - 8.23 (m; 2H, 2x -NH-)

b) Technetium-99m complex [9b] of the N- (3-hexadecylaminocarbonyl-2-acetylthiopropionyl) glycylglycylpropargylamide

A solution of 0.5 mg (0.9 μmol) of N- (3-hexadecylaminocarbonyl-2-acetylthiopropionyl) glycylglycylpropargylamide [9a] in 300 μl of DMSO is mixed with 30 μl of 1 N sodium hydroxide solution. Then 50 μl of a 0.15 molar trisodium citrate dihydrate solution and 0.4-0.9 mCi of a pertechnetate solution from a Mo-99 / Tc-99m generator are added. The reaction mixture is mixed in portions with 10 ul of a 0.2 molar tin (II) chloride dihydrate solution, incubated for 10 minutes at room temperature and then filtered (0.2 micron filter). The analysis of the label is carried out using HPLC: MERCK Nucleosil column, 125 x 4 mm, 5 μm; Gradient from 100% A to 100% B within 7.5 min; Eluent A: phosphate buffer (Na ₂ HP0; 0.01 M; pH 2.0); Eluent ^' B: acetonitrile / phosphate buffer (Na ₂ HP0; 0.01 M, pH 2.0) 50:50 (v / v); flow rate: 1.0 ml / min. The radiochemical purity of the Tc-99m complex is more than 95%. Example 10

a) N-dodecanoyl homocysteinylglycylglycylpropargylamide [10a]

A solution of 2.0 g (6.7 mmol) of N-dodecanoyl homocysteine thiolactone in anhydrous THF is mixed with a solution of 1.2 g (7.0 mmol) of glycylglycylpropargylamide in DMF. After the addition of 1 ml of triethylamine, the mixture is heated under reflux for 3 hours. The mixture is then concentrated under reduced pressure, the residue is taken up in CH ₂ C1 ₂ and washed with dilute HCl. The organic phases are washed neutral with water and dried over Mg ₂ SO ₄ . After the solvent has been concentrated under reduced pressure, it is recrystallized from methanol and, if appropriate, purified by column chromatography with CH ₂ Cl ₂ / MeOH (9: 1). Yield: 58% of the title compound [10a] C ₂₃ H ₄₀ N ₄ O ₄ S (MW: 468.66) ^** -H-NMR (dg-DMSO): δ = 0.86 (t; 3H, - (CH ₂ ) _g CH ₃ )

1.20 - 1.33 (m; 16H, - (CH ₂ ) gCH ₃ ) 1.48 - 1.57 (m; 2H, -CH ₂ -CH ₂ C0NH-) 2.10 - 2.19 (m ; 2H, -CH ₂ -CH ₂ CONH-) 2.56 - 2.73 (m; 2H, -CHCH ₂ CH ₂ SH) 3.10 (t; 1H, -C≡CH) 3.38 - 3, 44 (m; 2H, -CHCH ₂ CH ₂ SH) 3.52 (s; 1H, -SH)

3.65 - 3.73 (m; 4H, 2x -NH (CH ₂ ) CO-) 3.86 - 3.90 (m; 2H, -NH (CH ₂ ) -C≡CH) 4.48 - 4 , 54 (m; 1H, -CHCH ₂ CH ₂ SH) 8.05 - 8.10 (m; 2H, 2x -NH-) 8.24 (t; 1H, -NH-) 8.35 (t; 1H , -NH-) -> b) Technetium-99m complex [10b] of N-dodecanoylhomocysteinylglycylglycylpropargylamide

A solution of 0.5 mg (1.06 μmol) of N-dodecanoylhomo- 5 cysteinylglycylglycylpropargylamide [10a] in 300 μl of DMSO is mixed with 30 μl of 1 N sodium hydroxide solution. Then 50 μl of a 0.15 molar trisodium citrate dihydrate solution and 0.4-0.9 mCi of a pertechnetate solution from a Mo-99 / Tc-99m generator are added. 10 μl of a 0.2 molar tin (II) chloride dihydrate solution are added to the reaction mixture in portions, incubated for 10 minutes at room temperature and then filtered (0.2 μm filter). The analysis of the marking is carried out by means of HPLC: 5 MERCK Nucleosil column, 125 x 4 mm, 5 μm;

Gradient from 100% A to 100% B within 7.5 min; Eluent A: phosphate buffer (Na ₂ HP0; 0.01 M; pH 2.0); Eluent B: acetonitrile / phosphate buffer (Na HP0; 0.01 M, pH 2.0) 50:50 (v / v); Flow rate: 1.0 ml / min. 0 The radiochemical purity of the Tc-99m complex is more than 95%.

5 Example 11

a) N-decanoyl homocysteinylglycylglycylpropargylamide [11a]

A solution of 1.35 g (8.0 mmol) of glycylglycylpropargylamide in DMF is added to a solution of 2.0 g (7.4 mmol) of N-decanoyl homocysteine thiolactone in anhydrous THF. After the addition of 1 ml of triethylamine, the mixture is heated under reflux for 3 hours. The mixture is then concentrated under 5 reduced pressure, the residue in CH ₂ C1 taken up and washed with dilute HC1. The organic phases are washed neutral with water and dried over Mg ₂ SO ₄ . After concentration of the solvent under reduced pressure, the product is recrystallized from methanol and, if necessary, purified by column chromatography with CH ₂ Cl ₂ / MeOH (9: 1). Yield: 67% of the title compound [11a]

C ₂₁ H ₃ gN ₄ 0 ₄ S (MW: 440.61) ¹ H-NMR (dg-DMSO): δ = 0.85 (t; 3H, - (CH ₂ ) gCH ₃ )

1.18 - 1.30 (m; 12H, - (CH ₂ ) gCH ₃ ) 1.45 - 1.53 (m; 2H, -CH ₂ -CH ₂ CONH-) 2.11 - 2.22 (m ; 2H, -CH ₂ -CH ₂ C0NH-) 2.55 - 2.70 (m; 2H, -CHCH ₂ CH ₂ SH) 3.09 (t; 1H, -C≡CH)

3.37 - 3.42 (m; 2H, -CHCH ₂ CH ₂ SH) 3.50 (S; 1H, -SH)

3.65 - 3.75 (m; 4H, 2x -NH (CH ₂ ) C0-) 3.87 - 3.92 (m; 2H, -NH (CH ₂ ) -C≡CH) 4.48 - 4 , 53 (m; 1H, -CHCH ₂ CH ₂ SH) 8.08 - 8.12 (m; 2H, 2x -NH-) 8.22 (t; 1H, -NH-) 8.32 (t; 1H , -NH-)

b) Technetium-99m complex [11b] of N-decanoylhomocysteinylglycylglycylproparg lamid

A solution of 0.5 mg (1.1 μmol) of N-decanoylhomocysteineylglycylglycylpropargylamide [11a] in 300 μl DMSO is mixed with 30 μl 1 N sodium hydroxide solution. Then 50 μl of a 0.15 molar trisodium citrate dihydrate solution and 0.4-0.9 mCi of a pertechnetate solution from a Mo-99 / Tc-99m generator are added. The reaction mixture is mixed in portions with 10 ul of a 0.2 molar tin (II) chloride dihydrate solution, 10 minutes incubated at room temperature and then filtered

(0.2 μm filter).

The analysis of the label is carried out using HPLC: MERCK Nucleosil column, 125x4 mm, 5 μm; Gradient from 100% A to 100% B within 7.5 min, eluent A: phosphate buffer (Na ₂ HP0 ₄ ; 0.01 M; pH 2.0); Eluent B: acetonitrile / phosphate buffer (Na ₂ HP0; 0.01 M, pH 2.0) 50:50 (v / v); Flow rate: 1.0 ml / min. The radiochemical purity of the Tc-99m complex is more than 95%.

Example 12

a) N-Hexanoylhomocysteinylglycylglycylpropargylamid [12a]

A solution of 2.0 g (9.3 mmol) of N-hexanoyl homocysteine thiolactone in anhydrous THF is mixed with a solution of 1.7 g (10.0 mmol) of glycylglycylpropargylamide in DMF. After the addition of 1 ml of triethylamine, the mixture is heated under reflux for 3 hours. The mixture is then concentrated under reduced pressure, the residue is taken up in CH ₂ C1 ₂ and washed with dilute HCl. The organic phases are washed neutral with water and dried over Mg ₂ SO ₄ . After the solvent has been concentrated under reduced pressure, it is recrystallized from methanol and, if appropriate, purified by column chromatography with CH ₂ Cl ₂ / MeOH (9: 1). Yield: 60% of the title compound [12a]

C ₁₇ H ₂₈ N ₄ 0 ₄ S (MW: 384.50) ^• H NMR (dg-DMSO): δ = 0.86 (t; 3H, - (CH ₂ ) ₂ CH ₃ ) 1.20 - 1.31 (m; 12H, - (CH ₂ ) ₂ CH ₃ ) 1, 47 - 1, 54 (m; 2H, -CH ₂ -CH ₂ CONH-)

2nd 10-2.19 (m; 2H, -CH ₂ -CH ₂ CONH-) 2.56-2.88 (m; 2H, -CHCH ₂ CH ₂ SH)

3rd 11 (t; 1H, -C≡CH) 3.36 - 3.40 (m; 2H, -CHCH ₂ CH ₂ SH) 3.51 (s; 1H, -SH)

3.67 - 3.76 (m; 4H, 2x -NH (CH ₂ ) CO-) 3.85 - 3.89 (m; 2H, -NH (CH ₂ ) -C≡CH) 4.47 - 4 , 54 (m; 1H, -CHCH ₂ CH ₂ SH) 8.03 - 8.10 (m; 2H, 2x -NH-) 8.20 (t; 1H, -NH-) 8.29 (t; 1H , -NH-)

b) Technetium-99m complex [12b] of N-hexanoyl homocysteinylglycylglycylpropargylamide

A solution of 0.5 mg (1.3 μmol) N-hexanoylhomocysteineylglycylglycylpropargylamide [12a] in 300 μl DMSO is mixed with 30 μl 1 N sodium hydroxide solution. Then 50 μl of a 0.15 molar trisodium citrate dihydrate solution and 0.4-0.9 mCi of a pertechnetate solution from a Mo-99 / Tc-99m generator are added. The reaction mixture is added in portions with 10 ul of a 0.2 molar tin (II) chloride dihydrate solution, incubated for 10 minutes at room temperature and then filtered (0.2 micron filter).

The analysis of the marking is carried out by means of HPLC: MERCK Nucleosil column, 125 x 4 mm, 5 μm; Gradient from 100% A to 100% B within 7.5 min; Eluent A: phosphate buffer (Na ₂ HP0 ₄ ; 0.01 M; pH 2.0);

Eluent B: acetonitrile / phosphate buffer (Na ₂ HP0 ₄ ; 0.01 M, pH 2.0) 50:50 (v / v); Flow rate: 1.0 ml / min. The radiochemical purity of the Tc-99m complex is more than 95%. 46

Example 13

a) 3,17ß-Dihydroxy-17or- [5- (2-benzoylthioacetylglycylglycyl) amino-pent-l- (E) -en-3-in] -1,3,5-estratriene [13a]

A suspension of 120.0 mg (0.24 mmol) of 17β-hydroxy-17α-iodovinyl-1, 3,5-estratriene-3-tetrahydropyranyl ether, 0.3 g (0.85 mmol) of S-benzoylthioacetylglycylglycylpropargylamide [la ], 10.0 mg (0.044 mmol) benzyltriethylammonium chloride, 10.6 mg (9.2 μmol) tetrakis (triphenylphosphine) palladium (O) and 8.15 mg (0.043 mmol) copper (I) iodide in 4 ml of toluene is stirred for 72 hours at room temperature. Then water is added, the mixture is extracted twice with toluene and dried over Mg ₂ SO ₄ . After the solvent has been concentrated under reduced pressure, it is taken up in 5 ml of THF, 150 mg (0.6 mmol) of pyridinium-para-toluenesulfonic acid in 5 ml of ethanol are added and the mixture is heated under reflux for 3 h. The solvent is then concentrated under reduced pressure and the residue is purified by column chromatography with CH ₂ Cl ₂ / MeOH (8: 1). Yield: 41% of the title compound [13a] C ₃₆ H ₄₁ N ₃ 0gS (MW: 643.80) ¹ H-NMR (dg-DMSO): δ = 0.83 (s; 3H, -CH3 steroidal)

1.15 - 2.30 (m; 13H, steroidal) 2.65 - 2.74 (m; 2H, -CH ₂ - steroidal) 3.72 (d; 2H, -NH (CH ₂ ) CO-) 3 , 81 (d; 2H, -NH (CH ₂ ) CO-) 3.90 (S; 2H, -SCH ₂ CO-) 4.02 (d; 2H, -NH (CH ₂ ) -C≡C-) 5.65; 6.35 (AB; 2H, -CH = CH-) 6.43 (d; 1H, steroidal) 6.50 (dd; 1H, steroidal) 7.00 (d; 1H, steroidal)

7.44 - 7.49 (m; 2H, ArH) 7.51 - 7.56 (m; 1H, ArH)

7.90 - 7.96 (m; 2H, ArH) 8.23 (t; 1H, -NH-) 8.27 (t; 1H, -NH-) 8.80 (t; 1H, -NH-)

b) Technetium-9 m complex [13b] of 3,17ß-dihydroxy-17α- [5- (2-benzoylthioacetylglycylglycyl) aminopent-1- (E) -en-3-in] -1,3,5 - estratria

A solution of 0.5 mg (0.8 µmol) of 3,17β-dihydroxy-17o; - [5- (2-benzoylthioacetylglycylglycyl) amino-pent-1- (E) - en-3-in] -1.3 , 5-estratriene [13a] in 250 μl ethanol is mixed with 50 μl 1 N sodium hydroxide solution and incubated for 15 minutes at room temperature. Then 50 μl of a 0.15-molar trisodium citrate dihydrate solution and 2.5 μl of a 0.2-molar tin (II) chloride dihydrate solution are added. A pertechnetate solution (0.4-0.9 mCi) from a Mo-99 / Tc-99m generator is added to the reaction mixture, incubated for 10 minutes at room temperature and then filtered (0.2 μm filter) .

The analysis of the marking is carried out by means of HPLC: HAMILTON PRP-1 column, 125 x 4.6 mm, 5 μm; Gradient from 100% A to 100% B within 7.5 min; Eluent A: acetonitrile Eluent B: phosphate buffer (Na ₂ HP0 ₄ ; 0.001 M, pH 7.4); Flow rate: 2.0 ml / min.

The radiochemical purity of the Tc-99m complex is more than 95%. e

Example 14

a) 17ß-Hydroxy-17α- [5- (2-benzoylthioacetylglycylglycyl) amino-pent-l- (E, Z) -en-3-in] -4-estren-3-one [14a]

A suspension of 120.0 mg (0.28 mmol) of 17β-hydroxy-17α-iodovinyl-4-estren-3-one, 0.3 g (0.85 mmol) of S-benzoylthioacetylglycylglycylpropargylamide [la], 10, 0 mg (0.044 mmol) benzyltriethylammonium chloride, 10.6 mg (9.2 μmol) tetrakis (triphenylphosphine) palladium (0) and 8.15 mg (0.043 mmol) copper (I) iodide in 4 ml toluene are stirred for 72 hours at room temperature . Then water is added, the mixture is extracted twice with toluene and dried over Mg ₂ SO ₄ . After the solvent has been concentrated under reduced pressure, it is taken up in 5 ml of THF, 150 mg (0.6 mmol) of pyridinium-paratoluenesulfonic acid in 5 ml of ethanol are added and the mixture is heated under reflux for 3 hours. The solvent is then removed under reduced pressure and the residue is purified by column chromatography with CH ₂ Cl ₂ / MeOH (8: 1).

Yield: 39% of the title compound [14a] C ₃₆ H ₄₃ N ₃ 0 ₆ S (MW: 645.80)

- ^• -H-NMR (d -DMSO): δ = 0.95 (s; 3H, -CH ₃ steroidal)

0.82 - 2.50 (m; 20H, steroidal) 3.68 (d; 2H, -NH (CH ₂ ) C0-) 3.80 (d; 2H, -NH (CH ₂ ) C0-) 3, 87 (s; 2H, -SCH ₂ C0-) 3.94 - 3.98 (m; 2H, -NH (CH ₂ ) -C≡C-) 5, 60; 5.95 (AB; 2H, -CH = CH-) 5.82 (s; 1H, steroidal) 7.41 - 7.45 (m; 2H, ArH) 7, 49 - 7, 54 (m; 1H, ArH)

7, 85 - 7, 90 (m; 2H, ArH)

8, 24 (t; 1H, -NH-)

8, 29 (t; 1H, -NH-) 8,, 78 (t; 1H, -NH-)

b) Technetium-99m complex [14b] of 17ß-hydroxy-17α- [5- (2-benzoy1thioacetylglycylglycy1) a ino-pent-1- (E, Z) -en-3-in] -4-estren-3 -on

A solution of 0.5 mg (0.8 μmol) 17β-hydroxy-17α- [5- (2-benzoylthioacetylglycylglycyl) amino-pent-1- (E, Z) -en- 3-in] -4-estren- 3-one [14a] in 250 μl ethanol is mixed with 50 μl 1 N sodium hydroxide solution and incubated for 15 minutes at room temperature. Then you put 50 ul one

0.15-molar trisodium citrate dihydrate solution and 2.5 μl of a 0.2-molar tin (II) chloride dihydrate solution. The reaction mixture is mixed with a pertechnetate solution (0.4-0.9 mCi) from a Mo-99 / Tc-99m generator, incubated for 10 minutes at room temperature and then filtered (0.2 μm filter). The analysis of the marking is carried out by means of HPLC: HAMILTON PRP-1 column, 125 x 4.6 mm, 5 μm; Gradient from 100% A to 100% B within 7.5 min; Eluent A acetonitrile

Eluent B: phosphate buffer (Na ₂ HP0 ₄ ; 0.001 M; pH 7.4); Flow rate: 2.0 ml / min.

The radiochemical purity of the Tc-99m complex is more than 95%. Example 15

a) S-acetyl-thioacetylsarcosylglycylpropargylamide [15a]

A solution of 1.54 g (8.4 mmol) of sarcosylglycylpropargylamide in 30 ml of anhydrous THF is slowly mixed with a solution of 5.3 g (8.4 mmol) of S-acetylmercaptoacetic acid-N-hydroxysuccinimide ester in anhydrous THF . After 4 hours of stirring at room temperature, the mixture is heated to 40 ° C. for 30 minutes. After cooling, the solvent is concentrated under reduced pressure and the remaining residue is purified by column chromatography over silica gel [CH ₂ C1 ₂ / CH ₃ 0H (9: 1)]. The product crystallizes from ethanol.

Yield: 88% of the title compound [15a] C ₁₂ H ₁₇ N ₃ 0 ₄ S (MW: 299.35)

- ^• -H-NMR (dg -DMSO): δ = 2.34 (s; 3H, -COCH ₃ )

2.81 (S, 3H, -N-CH ₃₎

3.07 (t; 1H, -C≡CH)

3.70 (d; 2H, -NH (CH ₂ ) CO-)

3.82 (S; 2H, -NH (CH ₂ ) C0-)

3.86 - 3.90 (m; 2H, -NH (CH ₂ ) -C≡CH)

3.93 (S; 2H, -SCH ₂ C0-)

8.13 (t; 1H, -NH-)

8.22 (t; 1H, -NH-)

b) Technetium-99m complex [15b] of S-acetyl-thioacetylsarcosylglycylpropargylamide

A solution of 0.5 mg (1.67 μmol) S-acetyl-thioacetylsarcosylglycylpropargylamide [15a] in 50 μl 0.1 N sodium hydroxide solution is mixed with 250 μl phosphate buffer (Na ₂ HP0 ₄ , 0.5 mol / 1, pH 8.5). Then 50 μl of a 0.15-molar trisodium citrate dihydrate solution and 2.5 μl of a 0.2-molar tin (II) chloride dihydrate solution are added. A pertechnetate solution (0.4-0.9 mCi) from a Mo-99 / Tc-99m generator is added to the reaction mixture, incubated for 10 minutes at room temperature and then filtered (0.2 μm filter) . The analysis of the marking is carried out by means of HPLC: MERCK Nucleosil column, 125 x 4 mm, 5 μm; Gradient from 100% A to 100% B within 7.5 min, eluent A: phosphate buffer (Na ₂ HP0 ₄ ; 0.01 M; pH 2.0); Eluent B: acetonitrile / phosphate buffer (Na HP0; 0.01 M; pH 2.0) 50:50 (v / v); Flow rate: 1.0 ml / min. The radiochemical purity of the Tc-99m complex is more than 95%.

Example 16

a) 2- (S-acetylthio) succinylsarcosylglycylpropargyl amide [16a]

8.89 g (48.5 mmol) of sarcosylglycylpropargylamide are dissolved in 60 ml of anhydrous DMF under nitrogen, 8.5 g (48.7 mmol) of acetylmercapto-succinic anhydride are added and the mixture is stirred at 40 ° C. for 1 hour. After cooling, the product is precipitated with ether and centrifuged off. The crystalline product is recrystallized from THF.

Yield: 90% of the title compound [16a]

C ₁₄ H ₁₉ N ₃ 0 ₆ S (MW: 357.39) ¹ H-NMR (d -DMSO): δ = 2.34 (s; 3H, -C0CH ₃ ) 2.78 (ε; 3H, -N -CH3) 52

2.70 - 3.04 (m; 2H, - (CH ₂ ) COOH)

3.07 (t; 1H, -C≡CH)

3.68 - 3.73 (m; 4H, 2x -NH (CH ₂ ) CO-)

3.85 - 3.89 (m; 2H, -NH (CH ₂ ) -C≡CH)

4.33 - 4.38 (m; 1H, -S-CH (CH ₂ COOH) -CH ₂ -)

8.12 (t; 1H, -NH-)

8.23 (t; 1H, -NH-)

12.68 (broad; 1H, -COOH)

b) Technetium-99m complex [16b] of 2- (S-acetylthio) succinylsarcosylglycylpropargylamide

A solution of 0.5 mg (1.4 μmol) 2- (S-acetylthio) succinylsarcosylglycylpropargylamide [16a] in 50 μl 0.1 N sodium hydroxide solution is mixed with 250 μl phosphate buffer (Na ₂ HP0 ₄ , 0.5 mol / 1, pH 8.5). Then 50 μl of a 0.15 molar trisodium citrate dihydrate solution and 2.5 μl of a 0.2 molar tin (II) chloride dihydrate solution are added. A pertechnetate solution (0.4-0.9 mCi) from a Mo-99 / Tc-99m generator is added to the reaction mixture, incubated for 10 minutes at room temperature and then filtered (0.2 μm filter). The analysis of the marking is carried out by means of HPLC: MERCK Nucleosil column, 125 x 4 mm, 5 μm; Gradient from 100% A to 100% B within 7.5 min; Eluent A: phosphate buffer (Na ₂ HP0 ₄ ; 0.01 M; pH 2.0); Eluent B: acetonitrile / phosphate buffer (Na ₂ HP0 ₄ ; 0.01 M; pH 2.0) 50:50 (v / v); Flow rate: 1.0 ml / min.

The radiochemical purity of the Tc-99m complex is more than 95%. Example 17

a) (2- (S-acetylthio) succinylsarcosylglycylpropargylamid-4-yl) -Cys-Ser-Cys-Ser-Ser-Leu et-Asp-Lys-Glu-Cys-Val-Tyr-Phe-Cys-His -Leu-Asp-Ile-Ile-Trp [17a]

A solution of 50 mg of 2- (S-acetylthio) succinylsarcosylglycylpropargylamide [16a] and 15 mg of N-hydroxysuccinimide in anhydrous, freshly distilled DMF is cooled to - 15 ° C and mixed with 28 mg of dicyclohexylcarbodiimide in anhydrous DMF. The reaction mixture is stirred at -5 ° C. for 2 hours, then at room temperature for 2 hours and finally cooled to -15 ° C. Precipitated N, N'-dicyclohexylurea is filtered off. The filtrate is mixed with a solution of 1 mg Cys-Ser-Cys-Ser-Leu-Met-Asp-Lys-Glu-Cys-Val-Tyr-Phe-Cys-His-Leu-Asp-Ile-Ile-Trp (Endothelin 1) in anhydrous DMF and stirred for 20 hours at room temperature. The reaction solution is concentrated at room temperature under reduced pressure. After dropwise addition of diethyl ether, a flocculent precipitate is formed, which is centrifuged and purified by preparative HPLC (gradient: acetonitrile / phosphate buffer). After the buffered eluate has been neutralized, the organic solvent is blown off with nitrogen and the remaining residue is freeze-dried. The crystalline product is kept under protective gas (Ar). MG: calc. 2831.3 spec. 2831.6 (FAB-MS)

b) Technetium-99m complex [17b] des (2- (S-acetylthio) succinylsarcosylglycylpropargylamid-4-yl) Cys-Ser-Cys-Ser-Ser-Leu-Met-Asp-Lys-Glu-Cys-Val-Tyr-Phe-Cys-His-Leu-Asp-Ile-Ile-Trp

A solution of 0.5 mg (2- (S-acetylthio) succinylsar-cosylglycylpropargylamid-4-yl) -Cys-Ser-Cys-Ser-Ser-Leu-Met-Asp-Lys-Glu-Cys-Val-Tyr- Phe-Cys-His-Leu-Asp-Ile-Ile-Trp [17a] in 50 μl 0.1 N sodium hydroxide solution is mixed with 250 μl phosphate buffer (Na ₂ HP0 ₄ , 0.5 mol / 1, pH 8.5) ¬ thins. Then 50 μl of a 0.15 molar trisodium citrate dihydrate solution and 2.5 μl of a 0.2 molar tin (II) chloride dihydrate solution are added. The reaction mixture is mixed with a pertechnetate solution (0.4-0.9 mCi) from a Mo-99 / Tc-99m generator, incubated for 10 minutes at room temperature and then filtered (0.2 μm filter).

The analysis of the marking is carried out by means of HPLC: MERCK Nucleosil column, 125 x 4 mm, 5 μm; Gradient from 100% A to 100% B within 7.5 min, eluent A: phosphate buffer (Na ₂ HP0 ₄ ; 0.01 M; pH 2.0); Eluent B: acetonitrile / phosphate buffer (Na ₂ HP0 ₄ ; 0.01 M; pH 2.0) 50:50 (v / v); Flow rate: 1.0 ml / min. The radiochemical purity of the Tc-99m complex is more than 80%.

Example 18

a) Cyclo (Trp-Leu-Val-Pro-Asp) -Cys-Gly-Gly-Propargylamid [18a]

A solution of 133.0 mg cyclo (Trp-Leu-Val-Pro-Asp) and 41.3 mg hydroxybenzotriazole in 6 ml anhydrous, freshly distilled DMF is cooled to -15 ° C and with a solution of 51.7 mg l -Ethyl-3- (3-dimethylaminopropyl) carbodiimide hydrochloride in 8 ml of DMF. The Reaction mixture is stirred at -5 ° C for 2 hours. The mixture is stirred for a further 2 hours at room temperature and then a solution of 134.0 mg of S- (p-methoxybenzyl) cysteinylglycylglycylpropargylamide [5a] in anhydrous DMF is slowly added dropwise. After stirring for a further 7 hours, the solution is concentrated under reduced pressure and the peptide is precipitated with ether. The protective groups present are split off by stirring in liquid HF. After the HF has been evaporated off, the residue is purified by means of preparative HPLC (gradient: acetonitrile / phosphate buffer). After neutralization of the buffered eluate, the organic solvent portion is blown off with nitrogen and the remaining residue is freeze-dried. The crystalline product is stored under protective gas (Ar). MG: calc. 865.0 spec. 865.2 (FAB-MS)

b) Technetium-99m complex [18b] of cyclo (Trp-Leu-Val-Pro-Asp) -Cys-Gly-Gly-propargylamide

A solution of 0.5 mg cyclo (Trp-Leu-Val-Pro-Asp) -Cys-Gly-Gly-propargylamide [18a] in 300 μl phosphate buffer (Na ₂ HP0 ₄ , 0.5 mol / 1, pH 8, 5) 50 μl of a 0.15 molar trisodium citrate dihydrate solution and 2.5 μl of a * 0.2 molar tin (II) chloride dihydrate solution are added. A pertechnetate solution (0.4-0.9 mCi) from a Mo-99 / Tc-99m generator is added to the reaction mixture, incubated for 10 minutes at room temperature and then filtered (0.2 μm filter) . The analysis of the marking is carried out by means of HPLC: MERCK Nucleosil column, 125 x 4 mm, 5 μm; Gradient from 100% A to 100% B within 7.5 min; Eluent A: phosphate buffer (Na ₂ HP0 ₄ ; 0.01 M; pH 2.0); Eluent B: acetonitrile / phosphate buffer (Na HP0; 0.01 M; pH 2.0) 50:50 (v / v); Flow rate: 1.0 ml / min. The radiochemical purity of the Tc-99m complex is more than 90%.

Example 19

Enrichment of the N- (3-hexadecylaminocarbonyl-2-acetylthiopropionyl) glycylglycylpropargylamide, technetium-99m complex in atherosclerotic vascular lesions of WHHL rabbits

The N- (3-hexadecylaminocarbonyl-2-acetylthiopropionyl) glycylglycylpropargylamide (prepared according to Example 9a) is labeled as described in Example 9b.

99.9 GBq (2.7 mCi) of the substance labeled according to Example 9b was diluted to 1 ml with phosphate-buffered saline and applied to an anesthetized WHHL rabbit Rompun / Ketavet (1: 2) via an ear vein. The rabbit was sacrificed 5 hours after application and both an autoradiography of the aorta and a Sudan III staining to show the atherosclerotic

Plaques performed (Figure 1). The enrichment factor between normal and atherosclerotic wall areas was between 3 and 8, depending on the level of plaque (Sudan III staining).

Claims

 Claims

Compounds of the general formula (I),

characterized in that.

(A ¹ ), (A ² ) and (A ³ ) are the same or different and are for the general formula

are in which the two free valences are connected in any way with the respective nitrogen or sulfur atoms,

and in what

R ³ and R ^{5 are the} same or different, denote a hydrogen atom, a methyl or an ethyl group and R ^{4 represents} a hydrogen atom, a branched or unbranched alkyl group having 1-4 carbon atoms, the C atoms of which may have amino groups, NfR '-' - Rk) groups (where R ^a and R ^{* 0 are} identical or different and represent branched or unbranched alkyl or acyl radicals having 1-20 carbon atoms, the C atoms of which are substituted at most by one hydroxyl, one carboxy or one amino group), hydroxyl groups, thiol groups, halogen atoms, carboxy groups,

Alkoxycarbonyl groups with 1-20 carbon atoms, acyloxy groups with 1-12 carbon atoms, aminocarbonyl groups, sulfonyl groups, aminosulfonyl groups or phosphoric acid residues are substituted,

R ² and R ^{9 are the} same or different and have the same meaning as R ⁴ ,

k, 1 and m are the same or different and the numbers mean 0, 1, 2, 3 or 4 and

X is a hydrogen atom, a carboxy group, an alkoxy group with 1-20 carbon atoms, an alkoxycarbonyl group with 1-20 carbon atoms, an acyloxy group with 1-20 carbon atoms, an aminocarbonyl group, a sulfonyl group, an aminosulfonyl group, a phosphoric acid residue, a carboxymethylaminocarbonyl group, a p-ami - nophenyl group, a p-hydroxyphenyl group

Halogen atom, a hydroxyl group, an amino group, an N (R ^a R ^{* D} ) group (where R ^a and R ^{b are the} same or different and represent branched or unbranched alkyl or acyl radicals having 1-20 carbon atoms, whose C atoms, if appropriate with a hydroxy, a carboxy or a

Amino group are substituted), a hydrazine group, a hydrazide group, a cholesteryloxycarbonylmethylaminocarbonyl group, a cholesteryloxycarbonyl group, a cholesteryloxycarbonylmethyloxycarbonyl group, another

Steroid or a derivative of an ethynyl or

Ethenylsteroids, a substituent of the formula

_Z rl ^J -._ γ- ^i. _ (CH ₂ ) iQ-C0-

in which

Q is -NH- or -0-, Z ^{1 has} the same meaning as Z,

Y ^{1 has} the same meaning as Y and i have the same meaning as m,

a substituent of the formula

V U

in which

Q ^{1 is} a -NH-, -CO- or -0-, U is a bond, a group of the formula - (OC ^ CO) * ^ - with h = l-3 or a suitable linker for coupling to bio- or macromolecules and V is a hydrogen atom, a hydroxyl group, an NfR ⁶ ^ ^{** 3} ) group (where R ^a and R ^{** 3 are the} same or different and represent branched or unbranched alkyl or acyl radicals having 1-20 carbon atoms , whose C atoms are optionally substituted with a hydroxyl, a carboxy or an amino group), is a bio or macromolecule, means and Y is an unsaturated, at least one double and / or triple bond-containing, unbranched or branched chain with up to 12 carbon atoms, which may be one or more times and at any point in the chain with hydroxy, carboxy, alkoxy, amino or substituted amido groups with 1-20 carbon atoms in the alkyl and / or aryl radical means,

Z is a hydrogen atom, a halogen atom, a carboxy group, a hydroxy group, an alkoxycarbonyl group with 1-20 carbon atoms, an acyloxy group with 1-20 carbon atoms, an alkoxy group with 1-20 carbon atoms, one

Cholesteryloxycarbonylmethylaminocarbonyl group, a cholesteryloxycarbonyl group, a cholesteryloxycarbonylmethyloxycarbonyl group, another steroid or a derivative of an ethine or ethene steroid, a substituent of the formula

V U

in which

Q ^{1 is} a -NH-, -CO- or -0-,

U is a bond, a group of the formula

- (OCH ₂ C = 0) _jj - with h = l-3 or a suitable linker for coupling to bio or macromolecules and V is a hydrogen atom, a hydroxyl group, an N (R ^a R * °) group (where R ^a and R ^* ° are the same or different and represent branched or unbranched alkyl or acyl radicals having 1-20 carbon atoms, the C atoms of which may have a hydroxyl, a carboxy or an amino group are a bio- or macromolecule, means

M represents an element of atomic number 43 or 75,

R ^{1 has} the same meaning as R ⁴ ,

R ⁶ , R ⁷ and R ^{8 are the} same or different and are for a hydrogen atom, an alkyl group with 1-4 carbon atoms, the C atoms of which are optionally substituted with hydroxyl, carboxy or amino groups or for a group of the formula -CH ₂ -X, in which X has the meaning given above,

and their water-soluble salts

2. Compounds according to claim 1, characterized in that Z is a hydrogen atom, a steroid, an ethinyl steroid or ethenyl steroid.

3. Compounds according to claim 2, characterized in that X is a hydrogen atom, a halogen atom, a carboxy group, an amino group or an amido group with 1-20 carbon atoms in the alkyl and / or aryl radical.

Compounds according to claim 3, characterized gekennzeich¬ net that Y is an ethinylidene group, R ¹ , R ³ , R ⁶ , R ⁷ and R ^{8 are} hydrogen atoms and k, 1, and m is the number 0. 5. Compounds according to claim 1, characterized in that Z is a hydrogen atom and X is a cholesteryloxycarbonylmethylaminocarbonyl group, a cholesteryloxycarbonyl group, a cholesteryloxycarbonylmethyloxycarbonyl group, another steroid or a derivative of an ethynyl or ethenyl steroid.

6. Compounds according to claim 5, characterized gekennzeich¬ net that Y is an ethinylidene group, R ¹ , R ³ , R ^* *, R ⁷ and R ^{8 are} hydrogen atoms and k, 1, and m are the numbers 0 or 1.

Compounds according to claim 1, characterized gekennzeich¬ net that Z is a hydrogen atom and X is a hydrogen atom, a carboxy group or a substituent of the formula

V U

in which

Q ^{1 is} a -NH-, -CO- or -0-, U is a bond, a group of the formula - (OC ^ CO) - ^ - with h = l-3 or a suitable linker for coupling to bio or macromolecules and V is a hydrogen atom, a hydroxyl group, an N (R ^a R ^* °) group (where R ^a and R ^* ° are the same or different and represent branched or unbranched alkyl or acyl radicals having 1-20 carbon atoms , whose C atoms are optionally substituted with a hydroxyl, a carboxy or an amino group), is a bio or macromolecule. Compounds according to Claim 7, characterized in that Y is an ethinylidene group, R ¹ , R ³ , R ⁶ , R ⁷ and R ^{8 are} hydrogen atoms and k, 1 and m are the numbers 0, 1 or 2.

9. Compounds according to claim 1, characterized in that Z is a hydrogen atom and X is a

Hydrogen atom, a carboxy group, an alkoxy group with 1-20 carbon atoms, an alkoxycarbonyl group with 1-20 carbon atoms, an acyloxy group with 1-20 carbon atoms, an aminocarbonyl group, a sulfonyl group, an aminosulfonyl group, a phosphoric acid residue , a carboxymethylaminocarbonyl group, a p-aminophenyl group, a p-hydroxyphenyl group, a halogen radical, a hydroxy group, an amino group, an N (R ^a R ^{* b} ) group (where R ^a and R ^{b are the} same or different and are branched or unbranched alkyl or acyl radicals having 1-20 carbon atoms, the C atoms of which are optionally substituted by a hydroxyl, a carboxy or an amino group), one

Is hydrazine group or a hydrazide group.

10. Compounds according to claim 9, characterized in that Y is an ethinylidene group, R ¹ , R ³ , R ⁶ , R ⁷ and R ^{8 are} hydrogen atoms and k, 1 and m are the numbers 0, 1 or 2. 11. Compounds of the general formula (II),

characterized in that A ¹ , A ² , A ³ , R ¹ , R ⁶ , R ⁷ , R ⁸ , Y and Z have the meaning given in claim 1 and wherein

T is a hydrogen atom, an acetate group, a benzoate group, a p-methoxybenzyl group, an acetamidomethyl group, a benzamidomethyl group, a trimethylacetamidomethyl group, a hydroxyacetyl group or another suitable sulfur protecting group.

12. Conjugates containing compounds of the general formula I and / or II and substances which accumulate selectively in diseased tissues, there being a covalent bond between them and this in the case of substances containing amino groups such as peptides, proteins and antibodies or their fragments, amidic or in the case of substances containing hydroxyl groups such as fatty alcohols, ester-like or imidic in the case of substances containing aldehyde groups. 13. Conjugates according to claim 12, characterized in that the substances accumulating in the diseased tissue mean peptides, in particular endothelins, partial sequences of endothelins, endothelin analogs, endothelin derivatives or endothelin antagonists.

14. Conjugates according to claim 12, characterized in that the peptides contain the sequences or parts thereof

Cys-Ser-Cys-Ser-Ser-Leu-Met-Asp-Lys-Glu-Cys-Val * Tyr-Phe-Cys-His-Leu-Asp-Ile-Ile-Trp,

Cys-Ser-Cys-Ser-Ser-Trp-Leu-Asp-Lys-Glu-Cys-Val-Tyr-Phe-Cys-His-Leu-Asp-Ile-Ile-Trp,

Cγs-Thr-Cys-Phe-Thr-Tyr-Lys-Asp-Lys-Glu-Cys-Val-Tyr-Tyr-Cys-His-Leu-Asp-Ile-Ile-Trp,

Cys-Ser-Ala-Ser-Ser-Leu-Met-Asp-Lys-Glu-Ala-Val * '

Tyr-Phe-Cys-His-Leu-Asp-Ile-Ile-Trp,

I \ C s-Ser-Cys-Asn-Ser-Trp-Leu-Asp-Lys-Glu-Cys-Val-

Tyr-Phe-Cys-His-Leu-Asp-Ile-Ile-Trp,

Ala-Ser-Cys-Ser-Ser-Leu-Met-Asp-Lys-Glu-Cys-Val-Tyr-Phe-Ala-His-Leu-Asp-Ile-Ile-Trp,

Ala-Ser-Ala-Ser-Ser-Leu-Met-Asp-Lys-Glu-Ala-Val-Tyr-Phe-Ala-His-Leu-Asp-Ile-Ile-Trp, 66

Cys-Ser-Cys-Ser-Ser-Trp-Leu-Asp-Lys-Glu-Ala-Val-Tyr-Phe-Ala-His-Leu-Asp-Ile-Ile-Trp,

₎ j

Cys-Val-Tyr-Phe-Cys-His-Leu-Asp-Ile-Ile-Trp,

N-Acetyl-Leu-Met-Asp-Lys-Glu-Ala-Val-Tyr-Phe-Ala- His-Leu-Asp-Ile-Ile-Trp

the partial sequence

His-Leu-Asp-Ile-Ile-Trp,

or the cyclic amino acid sequences

Cyclo- (DTrp-DAsp-Pro-DVal-Leu),

Cyclo- (DGlu-Ala-alloDIle-Leu-DTrp)

exhibit.

15. A process for the preparation of the compounds of general formula (I), characterized in that technetium-99m or rhenium in the form of

Pertechnetate or perrhenate in the presence of a reducing agent and optionally an auxiliary ligand with a compound of the general formula (II)

wherein

A ¹ , A ² , A ³ , R ¹ , R ⁶ , R ⁷ , R ⁸ , Y and Z which in Claim 1 and T have the meaning given in Claim 11 is implemented.

16. Process for the preparation of the compounds of the general formula (II)

characterized in that a) a compound of the general formula (III)

wherein

A ¹ , A ² , A ³ , R ¹ , R ⁶ , R ⁷ , R ⁸ , Y and Z the in

Claim 1 have given meaning and a shark

Is halogen, with a compound of the general formula

T - S ^~ M

wherein M ^{+ is} an alkali metal cation and T has the meaning given in claim 11,

or b) a compound of the general formula

HN (R ⁶ ) -A ² -N (R ⁷ ) -A ³ -N (R ⁸ ) -YZ

wherein A ² , A ³ , R ⁶ , R ⁷ and R ^{8 have} the meaning given in claim 1 and with a compound of the general formula

T - S W

wherein A ¹ and T have the meaning given in claim 11 and W has the meaning of a leaving group which enables A ¹ with free amino c groups react.

17. Kit for the production of radiopharmaceuticals, consisting of a compound of the general formula (II) according to claim 11, a reducing agent and, if appropriate, an auxiliary ligand which are in the dry state or in solution, an instruction for use with a reaction instruction for implementation tion of the compounds described with Techne¬ tium-99m or rhenium in the form of a pertechnetate solution or perrhenate solution.

18. Radiopharmaceutical composition for the non-invasive in vivo presentation of receptors and receptor-containing tissue and / or atherosclerotic plaques, characterized in that it contains a compound according to claim 1, optionally with the additives customary in galenics, and that the compound according to claim 1 is prepared in a kit according to claim 17 with technetium-99m or rhenium in the form of a pertechnetate solution or perrhenate solution.

19. Radiopharmaceutical composition according to claim 18, characterized in that it contains a compound according to claim 1 in the form of liposomes and that the compound according to claim 1 in a kit according to claim 17 with technetium-99m or rhenium in the form of a pertechnetate solution or perrhenate solution is prepared.