DE4311021A1 - Bifunctional chelating agents, their technetium and rhenium complexes, processes for their preparation and radiopharmaceutical compositions containing these compounds - Google Patents

Description

The present invention relates to new technetium and Rhenium chelate compounds, process for their preparation lung and radiopharmaceutical containing these compounds table agents, as well as conjugates of these compounds selectively accumulating in diseased tissue Substances, especially peptides and proteins. The The invention further relates to the production of this connection funds containing funds and their use for radiodiagnostic examinations.

Radioactive metal ions have long been in the medical diagnostics and therapy applied. So gamma emitters such as the isotope Tc- 99m used for tumor detection. beta emitters like that Isotopes Re-186, Re-188 and Re-189 are found in the Tumor therapy application. That for nuclear medicine Questions most commonly used radionuclide is Technetium-99m, which due to its cheap physical properties (no corpuscular beam lung, 6 h physical half-life, 140 KeV gamma Radiation) and the resulting low radiation particularly good as a radioisotope for the Vivo diagnostics is suitable. Technetium-99m can be without problems from nuclide generators as pertechnetate win and is in this form directly for the manufacturer kits for routine clinical needs use.

The efficiency of radionuclides in in vivo diagnosis Stik, as well as the therapy depends on the specificity  and the selectivity of the labeled chelates to the target cell from. An improvement on these properties is by coupling the chelates to biomolecules after the To achieve "drug targeting" principle. As biomolecules there are antibodies, their fragments, hormones, Growth factors and substrates of receptors and Enzymes. So in the UK patent application GB 2,109,407 the use of radioactively labeled mono clonal antibody against tumor associated antigens for described the in vivo tumor diagnosis. Likewise, direct protein labeling via donor groups (amino, Amide, thiol,) of the protein (Rhodes, B.A. et al., J. Nukl. Med. 1986, 27, 685-693) or by insertion complexing agents (U.S. Patent 4,479,930 and Fritzberg, A.R. et al., J. Nucl. Med. 1986, 27, 957) with Techne tium-99m. These experimental methods however, are not available for clinical use Available because on the one hand the selectivity is too low and on the other hand the background activity in the organism is high to enable in vivo imaging.

The first receptor-binding radiopharmaceuticals were developed by J.P. DiZio et al. (J.Nucl. Med. 1992, 33; 558-569 and Biocon jugate Chem. 1991, 2; 353-366). An in vivo Imaging could not be realized with these connections become. The necessity proves to be a big disadvantage speed of extraction of the compounds described the marking medium. These conditions are not satisfactory because on the one hand the preparation of the Kit for the user with the smallest possible beam load must be carried out and on the other hand Suitable laboratories for implementation in only a few clinics of this work are available.  

Cyclame as described by Volkert et al. (Appl.Radiol.Isot. 1982, 33; 891-896) and Troutner et al. (J.Nucl.Med. 1980, 21; 443-448) are described, as well as N₂S₂- and N₃S systems show due to the lack of reactive Groups for linking to biomolecules or proteins little use per se as tissue-specific radiopharmaceutical kon (M. Borel et al., Appl.Radiat.Isot. 1992, 43, 425-436).

While for the marking Cyclame mentioned above as also known N₂O₂ systems (Pillai, M.R.A. et al .; Inorg. Chem. 1990, 29, 1850-1856) pH values of 10-13 are required, usually N₂S₂- and N₃S systems already at pH values of 9-11 xieren. For labeling labile conjugates of bio- and macromolecules these conditions are still too extreme, especially since some systems, such as L, L- Ethylene dicysteine (L, L-EC) only at pH values <10 (Verbruggen, A.M. et al .; J. Nucl. Med. 1992, 33, 551-557) have it marked sufficiently. The literaturbe knew MAG₃ even requires a temperature influence of 90-100 ° C for 10 minutes (Bannister, K.M. et al .; J. Nucl. Med. 1990, 31, 1568-1573) um in one for the clinical application of sufficient radiochemical purity to be manufactured. These conditions are for unsatisfactory routine clinical operation lend.

N₂S₂ and N₃S systems as described by G. Bormans et al. (Nucl. Med. Biol. 1990, 17, 499-506) and in the European patent EP 0 173 424 and EP 0 250 013 described meet the requirement sufficient stability of the corresponding technetium 99m complexes, but become too fast and without specific Enrichment excreted by the organism, so that this  only as kidney function diagnostics in the clinic Anwen Find and therefore a limited use possess, especially because increasingly Desire for specifically in diseased tissues enriching substances has increased.

To solve the refined nuclear medicine question positions there is an urgent need for stable bifunctional complexing agents, which the so far not show the disadvantages mentioned in the complexation and who are capable of conjugates with bio and macromo to form without their specificity and selectivity influence significantly. At the same time conditions for the application of these ver human relationships with regard to absorbed rays dose, stability and solubility of the compounds be fulfilled.

The invention is therefore based on the object To provide compounds and conjugates, the simplest possible process for their production to create and invent a kit wording of this compounds according to the invention / conjugates for their clinical To provide application. This task is accomplished by the present invention is accomplished.

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

characterized in which
(A¹), (A²) and (A³) are the same or different and are for the general formula

where the two free valences are connected in any way with the respective nitrogen or sulfur atoms,
and in what
R³ and R⁵ are the same or different, are a hydrogen atom or a methyl or an ethyl group and
R⁴ represents a hydrogen atom, a branched or unbranched alkyl group with 1-4 carbon atoms, the C atoms of which may be with amino groups, N (RaR ^b ) groups (where Ra and R ^{b are the} same or different and for branched or unbranched alkyl - or acyl radicals with 1-20 carbon atoms, the C atoms of which are optionally substituted with a hydroxyl, a carboxy or an amino group), hydroxyl groups, thiol groups, halogens, 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,
R² and R⁹ are the same or different and have the same meaning as R⁴,
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, one p-aminophenyl group, a p-hydroxyphenyl group, a halogen atom, a hydroxy group, an amino group, an N (RaR ^b ) group (where Ra and R ^{b are the} same or different and for branched or unbranched alkyl or acyl radicals with 1 -20 carbon atoms, the carbon atoms of which are optionally substituted with a hydroxyl, a carboxy or an amino group), a hydrazine group, a hydrazide group, a cholesteryloxycarbonylmethylaminocarbonyl group, a cholesteryloxycarbonyl group, a cholesteryloxycarbonylmethyloxycarbonyl group, another steroid or a derivative of one Ethinyl or ethenyl steroids,
a substituent of the formula

Z¹-Y¹- (CH₂) _i -Q-CO-

in which
Q is an -NH- or -O-,
Z¹ has the same meaning as Z,
y¹ has the same meaning as Y and
i have the same meaning as m,
a substituent of the formula

V-U-Q¹-

in which
Q¹ is -NH-, -CO- or -O-,
U is a bond, a group of the formula - (OCH₂CO) _h - with h = 1-3 or a suitable linker for coupling to bio or macromolecules and
V is a hydrogen atom, a hydroxyl group, an N (RaR ^b ) group (where Ra and R ^{b are the} same or different and represent branched or unbranched alkyl or acyl radicals with 1-20 carbon atoms, the C atoms of which may be , a carboxy or an amino group), a bio or macromolecule, means and
Y is an unsaturated, at least one double and / or triple bond-containing, unbranched or ver branched chain with up to 12 carbon atoms, which optionally 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 is substituted means
Z is a hydrogen atom, a halogen atom, a carboxy group, a hydroxy group, an alkoxycarbonyl group having 1-20 carbon atoms, an acyloxy group having 1-20 carbon atoms, an alkoxy group having 1-20 carbon atoms, a 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-Q¹-

in which
Q¹ is -NH-, -CO- or -O-,
U is a bond, a group of the formula - (OCH₂C = O) _h - with h = 1-3 or a suitable linker for coupling to bio or macromolecules and
V is a hydrogen atom, a hydroxyl group, an N (RaR ^b ) group (where Ra and R ^{b are the} same or different and represent branched or unbranched alkyl or acyl radicals with 1-20 carbon atoms, the C atoms of which may be , a carboxy or an amino group), a bio or macromolecule means,
M represents an element of atomic number 43 or 75,
R¹ has the same meaning as R⁴,
R⁶, R⁷ and R⁸ are identical or different and represent a hydrogen atom, an alkyl group with 1-4 carbon atoms, the C atoms of which are optionally substituted by hydroxy, carboxy or amino groups or represent a group of the formula -CH₂-X, in which X has the meaning given above
and their water-soluble salts.

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

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

Compounds according to the invention are also preferred of the general formula (I), which is characterized by net are that the radical Z is a hydrogen atom, a Steroid, an ethinyl 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 R¹, R³, R⁶, R⁷ and R⁸ each represent hydrogen atoms and k, 1 and in each stand for the number 0.

Another group of preferred according to the invention Compounds of general formula (I) is thereby characterized in that the radical Z is a hydrogen atom and X is cholesteryloxycarbonylmethylamino carbonyl group, a cholesteryloxycarbonyl group, a  Cholesteryloxycarbonylmethyloxycarbonyl group other steroid or a derivative of an ethine or Ethensteroids means.

Preferred compounds according to the invention are furthermore of the general formula (I), characterized in that Z is hydrogen and X is Cholesteryloxycarbonylmethylaminocarbonyl group, a Cholesteryloxycarbonyl group, a cholesteryloxycar bonylmethyloxycarbonylgruppe, another steroid or means a derivative of an ethine or ethene steroid, the radical Y represents an ethinylidene group and the R¹, R³, R⁶, R⁷ and R⁸ are each hydrogen atoms represent and k, 1 and m each for the number 0 stand.

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

V-U-Q¹-

is
in which
Q¹ is -NH-, -CO- or -O-,
U is a bond, a group of the formula - (OCH₂CO) _h - with h = 1-3 or a suitable linker for coupling to bio or macromolecules and
V is a hydrogen atom, a hydroxyl group, an N (RaR ^b ) group (where Ra and R ^{b are the} same or different and represent branched or unbranched alkyl or acyl radicals with 1-20 carbon atoms, the C atoms of which may be , a carboxy or an amino group), a bio or macromolecule.

Further preferred compounds of the invention General formula (I) are characterized in that the radical Z represents a hydrogen atom and the radical X a hydrogen atom, a carboxy group or one Substituents of the formula

V-U-Q¹-

is
in which
Q¹ is -NH-, -CO- or -O-,
U is a bond, a group of the formula - (OCH₂CO) _h - with h = 1-3 or a suitable linker for coupling to bio or macromolecules and V is a hydrogen atom, a hydroxyl group, an N (RaR ^b ) group (where Ra and R ^{b are the} same or different 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 bio or macromolecule ,
the radical Y represents an ethinylidene group and the radicals R¹, R³, R⁶, R⁷ and R⁸ each represent hydrogen atoms and k, 1 and m each represent the number 0.

A fourth 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, 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 (RaR ^b ) Group (where Ra and R ^{b are the} same or different 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), is 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 (RaR ^b ) - Group (where Ra and R ^{b are the} same or different 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 or represents a hydrazide group, the radical Y represents an ethinylidene group and the radicals R¹, R³, R⁶, R and R⁸ each represent hydrogen atoms, and k, 1 and m each represent the number 0th

The present invention further relates to compounds of the general formula (II)

wherein A¹, A², A³, R¹, R⁶, R⁷, R⁸, Y and Z are the above have the meaning given and in which T is water 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 represents a suitable sulfur protecting group.

The present invention also relates to Conjugates of general compounds of the invention my formulas I and II with themselves selectively in tissues enriching substances. Linking the fragments takes place depending on the type of to be connected Substances amidic, imidic or ester-like. Are the peptides accumulating in the tissues, Proteins, antibodies or fragments of these, so the link is made via their amino groups amidic; contain those that accumulate in tissues Substances hydroxy groups, so the link is made ester-like; contained in the tissues enriching substances aldehyde groups, so takes place Linking imidic.  

Those that accumulate in tissues are preferred Substances that accumulate in diseased tissues, especially in tissues where atheroskle red plaques are present.

Conjugates which are characterized thereby are also preferred records are that they accumulate in the diseased tissue substances containing peptides and proteins, in particular Endotheline, partial sequences of endothelins, Endothe lin analogs, endothelin derivatives or endothelin ant agonists mean.

This characterizes particularly preferred conjugates records that the peptides contain the sequences or parts from that

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 general formula (I)

takes place according to the methods known to the person skilled in the art, for example by reaction of technetium-99m in Form of pertechnetate that is made with little effort Molybdenum technetium generators can be eluted in Presence of a reducing agent (for example Sn (II) chloride or dithionide) and optionally one Auxiliary ligands (for example sodium citrate or natri umtartrate) with a compound of the general formula (II)

wherein
A¹, A², A³, R¹, R⁶, R⁷, R⁸, Y, Z and T have the meaning given above.

The reaction is preferably carried out in an aqueous medium Room temperature. The SH- Protecting group takes place in situ or according to the expert known literature methods, for example by basi hydrolysis, reductive cleavage, etc. (see on Example "Protective Groups in Organic Synthesis", T.W. Greene, John Wiley and Sons 1981).

The present invention also relates to Process for the preparation of the verb according to the invention of the general formula (II)

characterized in that one is known per se wise

• a) a compound of the general formula (III) wherein
  A¹, A², A³, R¹, R⁶, R⁷, R⁸, Y and Z have the meaning given above and Hal is a halogen, with a compound of the general formula T-SM⁺worin M⁺ means an alkali metal cation and T has the meaning given above ,
  or
• b) a compound of the general formula HN (R⁶) -A²-N (R⁷) -A³-N (R⁸) -Y-Zworin A², A³, R⁶, R⁷ and R⁸ have the meaning given above and
  with a compound of the general formula T-S-A¹-Wworin A¹ and T have the meaning given in Claim 11 and W has the meaning of a leaving group which enables A¹ to react with free amino groups,

implements.

The production of the required as starting substances N-haloacetylamides are carried out according to the literature Methods by acylation with an amino function Haloacetic acid halides (JACS, 1969, 90, 4508; Chem. Pharm. Bull. 1981, 29 (1), 128).

The required amines of the general formula

HN (R⁶) -A²-N (R⁷) -A³-N (R⁸) -Y-Z

are based on methods known from the literature of peptide syn these (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 α- and β-amino acids according to the Ver drive, for example via an addition / elimination Reaction of an amine (primary or secondary) with one  Carbonyl compound (for example acid chloride, ge mixed anhydride, activated ester) on N-unprotected coupled organic molecules. After splitting off the Ami Protection group according to known literature methods, at for example by acidic or basic hydrolysis, hydro genolysis, reductive elimination with alkali metals in liquid ammonia, in a second reaction remaining amino function again with an N-protected ten α- and β-amino acids according to known literature methods the (for example carbodiimide method) derivatized. The amino protective group is split off according to the literature methods mentioned above.

The general connections still required formula

T-S-A¹-W

are carried out by the methods known to the person skilled in the art Conversion of the carboxy group (s) corresponding carbon acids, for example according to the carbodiimide method (Fieser, Reagents for Organic Synthesis 10, 142) a mixed or cyclic anhydride (Org. Prep. Proc. Int. 1975, 7, 215) or an activated one Ester (Adv. Org. Chem. Part B, 472) synthesized.

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

The coupling options are of particular importance th of the chelating agents according to the invention, since they  are to be regarded as trifunctional and in addition to the metal Complexing both a coupling over the multiple bond as well as via a carboxy or amino group in Allow A¹, A² or A³.

Of particular interest according to the invention is the Provision of marking procedures under milder conditions than in previously known methods a rapid and quantitative implementation of the invention connections immediately before using the Enable radiopharmaceuticals.

Linking the chelated image is of great advantage ner about multiple binding with steroids, where choice as a coupling to a -CH₂ group of the steroid or a connection to the 17-ethynyl or 17- Ethenyl group of steroids becomes possible.

The affinity of some technetium is surprisingly good 99m labeled steroid derivatives to their respective Steroid hormone receptor. This is how the technetium complex shows of 3,17β-dihydroxy-17α- (5- [2-benzoylthioacetylglycyl glycyl] amino-pent-1- (E) -en-3-in) -1,3,5-estratriene (Example 13b) in the in vitro cytosol test (Carlson, K.E. et al. 1989, 32, 345-355) a receptor affinity corresponds to that of 17β-estradiol. This makes them suitable these compounds are excellent for the receptor ima went with Technetium-99m and to tumor diagnosis / therapy pie steroid hormone dependent tumors.

Another advantage of the present invention lies the fact that through the optional link over the carboxy or multiple bond the possibility provides solubility and pharmacokinetics of these complexes through chemical substitution at the level of the com  control plexer. Are of particular importance here biodegradable groups or linkers in X.

Connections with show surprising properties lipophilic substituents in X or Z, e.g. B. in the case the compound of Example 9 found in atheroskle enrich red plaques and thus a possibility to represent atherosclerotic changes show on vessel walls.

By choosing suitable bio and macromolecules (e.g. monoclonal antibodies, steroids,. . . ) in X or Z complexes according to the invention are obtained which have a Have surprisingly high tissue and organ specificity and therefore excellent for solving a series of dia gnostic and therapeutic problems can wear.

The complex-forming ligands of the gen my formula (II), in which Z denotes a hydrogen atom tet and / or ligands of the general formula (II), which contains an ethynyl function can according to the methods known to the person skilled in the art (for example R. Rossi et al., Tetrahedron 1983, 39, 287) on iodine vinyl be coupled steroids. The synthesis of the required Iodine vinyl steroids are produced using the method known from the literature (e.g. H. Hofmeister et al., Tetrahedron 1986, 42, 3575).

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

The coupling of the chelating agents to bio or macromoles Cooling takes place according to the methods known to the person skilled in the art by conversion of the carboxy group (s) on the chelating agent, for example according to the carbodiimide method (Fieser, Reagents for Organic Synthesis 10, 142) mixed or cyclic anhydride (Org. Prep. Proc. Int. 1975, 7, 215) or via an activated ester (Adv. Org. Chem. Part B, 472) and subsequent reak tion 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 or be linked to macromolecules, of which known is that they are in the organ to be examined, Particularly enrich the organ part or tissue. Such Molecules are, for example, enzymes, hormones, polysac charides such as dextrans or starches, porphyrins, bleomy cine, insulin, prostaglandins, steroid hormones, amino sugar, amino acids, peptides such as polylysine, proteins (such as immunoglobulins, monoclonal antibodies per, lectins), lipids (also in the form of liposomes) and DNA or RNA type nucleotides. Particularly noteworthy ben are conjugates with albumins such as human serum albu min, antibodies, such as monoclonal, for tumor-associated antigens specific antibodies or Antimyosin. Instead of biological macromolecules suitable synthetic polymers such as poly ethyleneimines, polyamides, ect. be tied up.

The pharmaceutical compositions formed therefrom are suitable for example for use in tumor diagnosis stik as well as tumor therapy, atherosclerosis diagnosis  stik or receptor imaging. The binding affinity and binding specificity of the pharmaceuticals formed Means for the target organ, target organ part or targetge Because of the conjugate formation, webe may or may not be slightly affected.

The production of the radiopharmaceuticals according to the invention means are also carried out according to known Way in which the complexing agents according to the invention and their conjugates - optionally with the addition of additives common in galenics - in an aqueous medium dissolves or suspended and then the solution or Suspension lyophilized if necessary. Suitable Additives are, for example, physiologically harmless Buffers (such as tromethamine), additions of Auxiliary ligands (such as sodium citrate or Sodium tartrate), reducing agents (such as Tin (II) chloride) or - if necessary - electrical lyte such as sodium chloride or - if required - (a) auxiliary common in galenics substance (s) (for example lactose, mannitol) and / or Surfactant (s) (for example lecithins, Tween®, Myrj®). The additives used must be in their composition a preparation of the compounds of the invention allow.

The invention also relates to a kit for Manufacture of radiopharmaceuticals, consisting of a Compound of general formula (II), a reducti onsmittel and optionally an auxiliary ligand, the be in a dry state or in solution, one Instructions for use with a reaction instruction for Implementation of the described connections with Techne tium-99m or rhenium in the form of a pertechnetate solution or perrhenate solution.  

The radiopharmaceutical agents according to the invention are preferred in an amount of 0.1 mCi to 50 mCi in an amount of 0.5 mCi to 30 mCi per 70 kg Body weight applied to a patient. Details of their Application and dosage are for example in "Radiotracers for Medical Applications", CRC-Press, Boca Raton, Florida. You are intra venous application determined. The invention Complex compounds / conjugates 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 agents according to the invention meet the diverse requirements for Suitability as radiopharmaceuticals for radio diagnostics and radiotherapy. So they are excellent at it suitable after i.v. Application in target fabrics enrich and thus enable a non-invasive Diagnosing appropriate tissues. The water solubility of the radiopharmaceutical agents according to the invention is - if necessary - by the usual in the galenics Chen auxiliaries guaranteed as described above. Furthermore, the radiopharmaceuticals according to the invention means not only high stability in vitro on, but also a surprisingly high stability in vivo, so that a release or an exchange of the im Radionuclide bound complex or not clinically not relevant.

Preferred radiopharmaceutical compositions according to the invention Settings are also characterized in that the compounds of the general invention Formula I or II contains in the form of liposomes and that  optionally the compound of the invention common formula (I) in a kit with technetium or Rhenium is prepared in the form of permetallates.

The radiopharmaceutical agents according to the invention can also be used in radioimmuno or radiation therapy be used. These differ from the Radio diagnosis only by the amount and type of use radioisotopes. The goal is to destroy Tumor cells from high-energy short-wave radiation with the shortest possible range. Suitable β- emitting isotopes are, for example, rhenium-186 and Rhenium-188.

The radiopharmaceutical agents according to the invention are for the non-invasive in vivo presentation of steroids tissues containing zeptor, for example steroid hormone pending tumors.

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

Overall, new complexing agents and Synthesizing metal complexes is a new possibility in diagnostic and therapeutic medicine open up. Above all, the development of a new kind of image best practice in nuclear medicine diagnostics Stik makes this development appear desirable.  

The following examples illustrate the invention.

example 1 a) S-benzoylthioacetylglycylglycylpropargylamide [1a]

A solution of 150 mg (0.61 mmol) chloroacetylglycylglycylpropargylamide in water free DMF a solution of 215.1 mg (1.22 mmol) of potassium thiobenzoate added dropwise in anhydrous DMF. Then Catalytic amounts of sodium iodide are added and heated to 110 ° C for 2 hours. After cooling the Reaction mixture at room temperature is in 1 N HCl stirred in and extracted with ethyl acetate until there is no longer any product in the water phase. The organic phases are up to under reduced pressure evaporated to dryness, again taken up in ethyl acetate men, washed with water and dried over Mg₂SO₄. After the solvent has been stripped off under reduced pressure Pressure is recrystallized from methanol or if required by column chromatography with CH₂Cl₂ / MeOH (9: 1) cleaned.

Yield: 76% of the title compound [1a]
C₁₆H₁₇N₃O₄S (MG: 347, 39)
1 H-NMR (d₆-DMSO):
δ = 3.11 (t; 1H, -C≡CH)
3.69 (d; 2H, -NH (CH₂) CO-)
3.78 (d; 2H, -NH (CH₂) CO-)
3.84-3.88 (m; 2H, -NH (CH₂) -C≡CH)
3.90 (s; 2H, -SCH₂CO-)
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 [1b] of S-benzoylthioace tylglycylglycylpropargylamide

A solution of 0.5 mg (1.44 µmol) S-benzoylthioacetyl glycylglycylpropargylamide [1a] in 50 µl 1 N sodium hydroxide solution is mixed with 250 µl phosphate buffer (Na₂HPO₄, 0.5 mol / l, pH 8.5). Then you put 50 ul one 0.15 molar trisodium citrate dihydrate solution and 2.5 µl a 0.2 molar tin (II) chloride dihydrate solution. The reaction mixture is with a pertechnetate solution (0.4-0.9 mCi) from a Mo-99 / Tc-99m generator ver sets, incubated at room temperature for 10 minutes 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 (Na₂HPO₄; 0.01 M; pH 2.0); Eluent B: acetonitrile / phosphate buffer (Na₂HPO₄; 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)

5.89 g (37.5 mmol) of glycylgly cylpropargylamide dissolved in 60 ml of anhydrous DMF, with 8.50 g (48.7 mmol) of acetylmercapto-succinic anhydride  added and stirred at room temperature overnight. The reaction mixture is then reduced reduced pressure to dryness. The backlog is in Suspended methanol, suction filtered and umkri from water installed.

Yield: 61% of the title compound [2a] C₁₃H₁₇N₃O₆S (MW: 343.36)
1 H-NMR (d₆-DMSO):
δ = 2.35 (s; 3H, -COCH₃)
2.66; 2.83 (2x dd; 2H, - (CH₂) COOH)
2.99 (t; 1H, -C≡CH)
3.70-3.77 (m; 4H, 2x -NH (CH₂) CO-)
3.88 (dd; 2H, -NH (CH₂) -C≡CH)
4.35 (t; 1H, -S-CH (CH₂COOH) -CH₂-)
8.07 (t; 1H, -NH-)
8.12 (t; 1H, -NH-)
8.26 (t; 1H, -NH-)

b) Technetium-99m complex [2b) of 2-acetylthiosucci nylglycylglycylpropargylamide

A solution of 0.5 mg (1.45 µmol) 2-acetylthiosucci nylglycylglycylpropargylamid [2a] in 50 µl 0.1 N Sodium hydroxide solution is mixed with 250 µl phosphate buffer (Na₂HPO₄, 0.5 mol / l, pH 8.5). Then you set 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 too. The reaction mixture is with a Pertech netate solution (0.4-0.9 mCi) from a Mo-99 / Tc-99m gene rator added, 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 (Na₂HPO₄; 0.01 M; pH 2.0); Eluent B: acetonitrile / phosphate buffer (Na₂HPO₄; 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-thioacetylglutaminylglycylpropargylamide [3a]

A solution / suspension of 2.0 g (8.3 mmol) glutaminyl glycylpropargylamide in 80 ml of anhydrous DMF slowly with a solution of 5.3 g (8.4 mmol) of S-acetyl mercaptoacetic acid N-hydroxysuccinimide ester in water free THF. After stirring for 4 hours at room temperature temperature is hydrolyzed with water. Then will the solvent is removed under reduced pressure and the residue recrystallized from methanol / water.

Yield: 45% of the title compound [3a) C₁₄H₁₉N₃O₆S (MW: 357.36)
1 H-NMR (d₆-DMSO):
δ = 1.64-1.92 (m; 2H, -CHCH₂CH₂COOH)
2.31 (s; 3H, -COCH₃)
2.36 (t; 2H, -CHCH₂CH₂COOH)
3.02 (t; 1H, -C≡CH)
3.70 (s; 2H, -NH (CH₂) CO-)
3.73-3.80 (m; 1H, -CHCH₂CH₂COOH)
3.89 (dd; 2H, -NH (CH₂) -C≡CH)
4.21 (s; 2H, -SCH₂CO-)
8.09 (m; 1H, -NH-)
8.19 (m; 1H, -NH-)
8.26 (m; 1H, -NH-)

b) Technetium-99m complex [3b] of S-acetyl-thioace tylglutaminylglycylpropargylamide

A solution of 0.5 mg (1.40 µmol) S-acetylthioacetyl glutaminylglycylpropargylamide [3b] in 50 µl 0.1 N Sodium hydroxide solution is mixed with 250 µl phosphate buffer (Na₂HPO₄₁, 0.5 mol / l, pH 8.5). Then you set 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 too. The reaction mixture is with a Pertech netate solution (0.4-0.9 mCi) from a Mo-99 / Tc-99m gene rator added, 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 (Na₂HPO₄; 0.01 M; pH 2.0); Eluent B: acetonitrile / phosphate buffer (Na₂HPO₄; 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) glycyl glutaminylpropargylamide in 80 ml of anhydrous DMF slowly with a solution of 5.3 g (8.4 mmol) of S-acetyl mercaptoacetic acid N-hydroxysuccinimide ester in water  free THF. After stirring for 4 hours at room temperature temperature is hydrolyzed with water. Then will the solvent is removed under reduced pressure and the residue recrystallized from methanol / water.

Yield: 61% of the title compound [4a]
C₁₄H₁₉N₃O₆S (MG: 357.36)
1 H-NMR (d₆-DMSO):
δ = 1.60-1.88 (m; 2H, -CHCH₂CH₂COOH)
2.30 (s; 3H, -COCH₃)
2.28 (t; 2H, -CHCH₂CH₂COOH)
3.04 (t; 1H, -C≡CH)
3.68 (s; 2H, -NH (CH₂) CO-)
3.70-3.77 (m; 1H, -CHCH₂CH₂COOH)
3.88 (dd; 2H, -NH (CH₂) -C≡CH)
4.19 (s; 2H, -SCH₂CO-)
8.11 (m; 1H, -NH-)
8.23 (m; 1H, -NH-)
8.35 (m; 1H, -NH-)

b) Technetium-99m complex [4b] of S-acetyl-thioace tylglycylglutaminylpropargylamide

A solution of 0.5 mg (1.40 µmol) S-acetylthioacetyl glycylglutaminylpropargylamide [4a] in 50 µl 0.1 N Sodium hydroxide solution is mixed with 250 µl phosphate buffer (Na₂HPO₄, 0.5 mol / l, pH 7.5). Then you set 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 too. The reaction mixture is with a Pertech netate solution (0.4-0.9 mCi) from a Mo-99 / Tc-99m gene rator added, 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 (Na₂HPO₄; 0.01 M; pH 2.0); Eluent B: acetonitrile / phosphate buffer (Na₂HPO₄; 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) cysteinylglycylglycyl propargylamide [5a]

9.8 g (0.02 mol) of N-tert-butoxycarbonyl-S- (p-methoxy benzyl) cysteinylglycylglycylpropargylamid are at 0 ° C dissolved in trifluoroacetic acid and 4 hours in Ice bath stirred. Then it is reduced Pressure concentrated to dryness, the residue with ethanol added and concentrated again. The backlog is through Column chromatography on silica gel [CH₂Cl₂ / CH₃OH (5: 1)] cleaned. The product crystallizes after Constrict.

Yield: 93% of the title compound [5a]
C₁₈H₂₄N₄O₄S (MG: 392.48)
1 H-NMR (CD₃OD):
δ = 2.54 (t; 1H, -C≡CH)
2.63-2.86 (m; 2H, -CHCH₂S-)
3.48-3.52 (m; 1H, -CHCH₂S-)
3.71 (s; 2H, -NH (CH₂) CO-)
3.77 (s; 3H, -OCH₃)
3.85 (s; 2H, -NH (CH₂) CO-)
3.92 (s; 2H, -CH₂Ar)
3.97 (dd; 2H, -NH (CH₂) -C≡CH)
6.86; 7.25 (AA′BB ′; 4H, ArH)

b) Technetium-99m complex [5b] of cysteinylglycyl glycylpropargylamide

A solution of 0.5 mg (1.27 µmol) S- (p-methoxy benzyl) cysteinylglycylglycylpropargylamide [5a] in liquid HF is stirred for 15 minutes at 0 ° C. After this Evaporation of the solvent at room temperature the residue was taken up in 50 μl of 1 N sodium hydroxide solution and with 250 µl phosphate buffer (Na₂HPO₄₁ 0.5 mol / l, pH 7.5) diluted. Then 50 µl of 0.15 is placed molar trisodium citrate dihydrate solution and 2.5 µl a 0.2 molar tin (II) chloride dihydrate solution. The reaction mixture is with a pertechnetate solution (0.4-0.9 mCi) from a Mo-99 / Tc-99m generator added, 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 (Na₂HPO₄; 0.01 M; pH 2.0); Eluent B: acetonitrile / phosphate buffer (Na₂HPO₄; 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-propio nyl) -glycylglycylpropargylamid [6a]

A solution of 5.0 g (8.9 mmol) of 3-acetylthio amber Acid cholesterol ester in anhydrous THF is in the Chilled ice bath and dry with triethylamine in excess shot. Then 1.9 g (17.8 mmol)  Ethyl chloroformate added dropwise. You let go Warm to 0 ° C, mixed with 3.0 g (18.0 mmol) glycylgly cylpropargylamide and stirred for 3 hours at room temperature door. The reaction mixture is hydrolyzed with water and extracted with ethyl acetate until no product is more in the water phase. The organic Phases are dried to dryness under reduced pressure concentrated, again taken up in ethyl acetate, with water washed and dried over Mg₂SO₄. After removal the solvent under reduced pressure recrystallized from methanol or optionally using column chromatography with CH₂Cl₂ / MeOH (9: 1) cleaned.

Yield: 67% of the title compound [6a]
C₄₀H₆₁N₃O₆S (MG: 712.01)
1 H-NMR (CDCl₃):
d = 0.68-2.08 (m; 41H, steroidal)
2.30 (d; 2H, -C = CHCH₂- steroidal)
2.32 (s; 3H, -COCH₃)
2.76-3.00 (m; 2H, - (CH₂) COO-)
3.10 (t, 1H, -C≡CH)
3.66-3.73 (m; 4H, 2x -NH (CH₂) CO-)
3.84 (d; 2H, -NH (CH₂) -C≡CH)
4.39 (t; 1H, -S-CH (CH₂COOH) -CH₂-)
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] glycylglycylpro pargylamide

A solution of 0.5 mg (0.7 µmol) of N- [2-acetylthio-3- cholesteryloxycarbonylpropionyl] glycylglycylpropar gylamid [6a] in 300 µl DMSO with 30 µl 1 N sodium lye mixed. Then 50 µl of 0.15 is placed molar trisodium citrate dihydrate solution and 0.4-0.9 mCi a pertechnetate solution from a Mo-99 / Tc-99m genera gate to. The reaction mixture is added in portions with 10 µl of a 0.2 molar tin (II) chloride dihydrate solution added, 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 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) Cholesterol [N- (2-acetylthiosuccinylglycylglycyl propargylamid-4-yl) glycine] ester (7a)

A solution of 2.0 g (5.8 mmol) of 2-acetylthiosuccinyl glycylglycylpropargylamid [2a] in anhydrous THF cooled in an ice bath and with dry triethylamine in Excess offset. Then 1.3 g (11.6 mmol) ethyl chloroformate was added dropwise. You leave Warm to 0 ° C, add 5.0 g (12.0 mmol) Gly  cincholesterol ester and stir for 3 hours at room temperature maturity. The reaction mixture is hydrolyzed with water and extracted with ethyl acetate until none Product is more in the water phase. The organic phases are washed with water and over Mg₂SO₄ dried. After removing the solution with tels under reduced pressure is umkri from methanol installed or, if necessary, by means of column chromatography graph with CH₂Cl₂ / MeOH (9: 1) cleaned.

Yield: 41% of the title compound [7a]
C₄₂H₆₄N₃O₆S (MG: 769.06)
1 H-NMR (CD₃OD):
δ = 0.68-2.10 (m; 41H, steroidal)
2.32 (d; 2H, -C = CHCH₂- steroidal)
2.33 (s; 3H, -COCH₃)
2.81-3.10 (m; 2H, -CH (CH₂) COO-)
3.08 (t; 1H, -C≡CH)
3.41 (d; 2H, -NH (CH₂) CO-)
3.64-3.71 (m; 4H, 2x -NH (CH₂) CO-)
3.83 (d; 2H, -NH (CH₂) -C≡CH)
4.35 (t; 1H, -S-CH (CH₂COOH) -CH₂-)
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 cholesterol [N- (2- acetylthio-succinylglycylglycylpropargylamid-4- yl) glycine] esters

A solution of 0.5 mg (0.65 µmol) cholesterol [N- (2- Acetylthiosuccinylglycylglycylpropargylamid-4-yl) -gly  cin] ester [7a] in 300 µl DMSO is mixed with 30 µl 0.1 N Sodium hydroxide solution added. Then put 50 µl 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 too. The reaction mixture becomes por with 10 µl of a 0.2 molar tin (II) chloride Dihydrate solution added, 10 minutes at room temperature incubated and then filtered (0.2 µm filter).

The analysis of the marking is done by 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) cysteinylglycyl glycylpropargylamide [8a]

2.0 g (5.1 mmol) of S- (p-methoxybenzyl) cysteinylglycyl glycylpropargylamide [5a] are dissolved in CH₂Cl₂ and with little NEt₃ added. Then 1.2 g (5.5 mmol) of dodecanoic acid chloride in CH₂Cl₂ and 2 Stirred for hours at room temperature. The reaction ge mix is hydrolyzed with water and with CH₂Cl₂ extracted. After drying is reduced Concentrated pressure and the residue with CH₂Cl₂ / MeOH (9: 1) Chromatographed on silica gel.

Yield: 80% of the title compound [8a]
C₃₀H₄₆N₄O₅S (MG: 574.79)
1 H-NMR (d₆-DMSO):
δ = 0.85 (t; 3H, - (CH₂) ₈CH₃)
1.19-1.30 (m; 16H, - (CH₂) ₈CH₃)
1.46-1.54 (m; 2H, -CH₂-CH₂CONH-)
2.08-2.20 (m; 2H, -CH₂-CH₂CONH-)
2.51-2.79 (m; 2H, -CHCH₂S-)
3.08 (t; 1H, -C≡CH)
3.74 (s; 3H, -OCH₃)
3.67-3.78 (m; 4H, 2x -NH (CH₂) CO-)
3.76 (d; 2H, -CH₂Ar)
3.85-3.88 (m; 2H, -NH (CH₂) -C≡CH)
4.49-4.55 (m; 1H, -CHCH₂S-)
6.85; 7.23 (AA′BB ′; 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) N-dodecanoyl-cystei nylglycylglycylpropargylamid [8a] in liquid HF Stirred at 0 ° C for 15 min. After evaporation of the solution at room temperature the residue is dissolved in 300 µl DMSO dissolved and mixed with 30 ul 1 N sodium hydroxide solution. Then 50 µl of a 0.15 molar trina is placed trium citrate dihydrate solution and 0.4-0.9 mCi per technetate solution from a Mo-99 / Tc-99m generator. The reaction mixture is portioned with 10 ul one 0.2 molar tin (II) chloride dihydrate solution added, Incubate for 10 minutes at room temperature and then filtered down (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 (Na₂HPO₄; 0.01 M; pH 2.0); Eluent B: acetonitrile / phosphate buffer (Na₂HPO₄; 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-propio nyl) glycylglycylpropargylamide [9a]

A solution of 2.0 g (5.8 mmol) of 2-acetylthiosuccinyl glycylglycylpropargylamid [2a] in anhydrous THF cooled in an ice bath and with 1.7 g (7.0 mmol) of hexa decylamine added. Then 1.47 g at 0 ° C (7.0 mmol) dicyclohexylcarbodiimide in anhydrous THF, which was mixed with 2 ml of triethylamine was added dropwise. The mixture is allowed to warm to room temperature and stirred for 3 hours at this temperature. The reaction mixture is mixed with a few drops of acetic acid, with water hydrolyzed and extracted with ethyl acetate until there is no longer any product in the water phase. The organic phases are washed with water and over Mg₂SO₄ dried. After concentrating the solvent is recrystallized from methanol under reduced pressure siert and optionally by column chromatography cleaned with CH₂Cl₂ / MeOH (9: 1).

Yield: 69% of the title compound [9a]
C₂₉H₅₀N₄O₅S (MG: 566.81)
1 H-NMR (d₆-DMSO):
δ = 0.87 (t; 3H, -CH₂CH₃)
1.16-1.35 (m; 28H, - (CH₂) ₁₄CH₃)
2.33 (s; 3H, -COCH₃)
2.73-3.10 (m; 4H, - (CH₂) -)
3.07 (t; 1H, -C≡CH)
3.67-3.76 (m; 4H, 2x -NH (CH₂) CO-)
3.85-3.89 (m; 2H, -NH (CH₂) -C≡CH)
4.29-4.35 (m; 1H, -S-CH (CH₂COOH) -CH₂-)
7.99 (t; 1H, -NH-)
8.07 (t; 1H, -NH-)
8.15-8.23 (m; 2H, 2x -NH-)

b) Technetium-99m complex [9b] of N- (3-hexadecyl aminocarbonyl-2-acetylthiopropionyl) glycyl glycylpropargylamide

A solution of 0.5 mg (0.9 µmol) of N- (3-hexadecylamino carbonyl-2-acetylthiopropionyl) glycylglycylpropar gylamid [9a] in 300 µl DMSO with 30 µl 1 N sodium lye mixed. Then 50 µl of 0.15 is placed molar trisodium citrate dihydrate solution and 0.4-0.9 mCi a pertechnetate solution from a Mo-99 / Tc-99m genera gate to. The reaction mixture is added in portions with 10 µl of a 0.2 molar tin (II) chloride dihydrate solution added, 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 (Na₂HPO₄; 0.01 M; pH 2.0); Eluent B: acetonitrile / phosphate buffer (Na₂HPO₄; 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-Dodecanoylhomocysteinylglycylglycylpropargylamid [10a]

A solution of 2.0 g (6.7 mmol) of N-dodecanoylhomo cysteine thiolactone in anhydrous THF is mixed with a Solution of 1.2 g (7.0 mmol) of glycylglycylpropargylamide put in DMF. After adding 1 ml of triethylamine is heated under reflux for 3 hours. Then will concentrated under reduced pressure, the residue in CH₂Cl₂ added and washed with dilute HCl. The organic phases are washed neutral with water and dried over Mg₂SO₄. After narrowing the Solvent under reduced pressure becomes metha nol recrystallized and optionally by columns Chromatography with CH₂Cl₂ / MeOH (9: 1) cleaned.

Yield: 58% of the title compound [10a]
C₂₃H₄₀N₄O₄S (MG: 468.66)
1 H-NMR (d₆-DMSO):
δ = 0.86 (t; 3H, - (CH₂) ₈CH₃)
1.20-1.33 (m; 16H, - (CH₂) ₈CH₃)
1.48-1.57 (m; 2H, -CH₂-CH₂CONH-)
2.10-2.19 (m; 2H, -CH₂-CH₂CONH-)
2.56-2.73 (m; 2H, -CHCH₂CH₂SH)
3.10 (t; 1H, -C≡CH)
3.38-3.44 (m; 2H, -CHCH₂CH₂SH)
3.52 (s; 1H, -SH)
3.65-3.73 (m; 4H, 2x -NH (CH₂) CO-)
3.86-3.90 (m; 2H, -NH (CH₂) -C≡CH)
4.48-4.54 (m; 1H, -CHCH₂CH₂SH)
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-dodecanoylho mocysteinylglycylglycylpropargylamide

A solution of 0.5 mg (1.06 µmol) of N-dodecanoylhomo cysteinylglycylglycylpropargylamid [10a] in 300 µl DMSO is mixed with 30 ul 1 N sodium hydroxide solution. Subsequently put 50 µl of a 0.15 molar trisodium citrate di hydrate solution and 0.4-0.9 mCi of a pertechnetate solution from a Mo-99 / Tc-99m generator too. The reaction ge is mixed in portions with 10 ul 0.2 molar Tin (II) chloride dihydrate solution added, 10 minutes incubated 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 (Na₂HPO₄; 0.01 M; pH 2.0); Eluent B: acetonitrile / phosphate buffer (Na₂HPO₄; 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 11 a) N-Decanoylhomocysteinylglycylglycylpropargylamid [11a]

A solution of 2.0 g (7.4 mmol) of N-decanoyl homocysteine thiolactone in anhydrous THF comes with a solution of 1.35 g (8.0 mmol) glycylglycylpropargylamide in DMF transferred. After adding 1 ml of triethylamine, 3 Heated under reflux for hours. Then under concentrated pressure, the residue in CH₂Cl₂  taken up and washed with dilute HCl. The orga African phases are washed neutral with water and dried over Mg₂SO₄. After narrowing the solution by means of reduced pressure from methanol recrystallized and optionally by column chromatography topography with CH₂Cl₂ / MeOH (9: 1) cleaned.

Yield: 67% of the title compound [11a]
C₂₁H₃₆N₄O₄S (MG: 440.61)
1 H-NMR (d₆-DMSO):
δ = 0.85 (t; 3H, - (CH₂) ₆CH₃)
1.18-1.30 (m; 12H, - (CH₂) ₆CH₃)
1.45-1.53 (m; 2H, -CH₂-CH₂CONH-)
2.11-2.22 (m; 2H, -CH₂-CH₂CONH-)
2.55-2.70 (m; 2H, -CHCH₂CH₂SH)
3.09 (t; 1H, -C≡CH)
3.37-3.42 (m; 2H, -CHCH₂CH₂SH)
3.50 (s; 1H, -SH)
3.65-3.75 (m; 4H, 2x -NH (CH₂) CO-)
3.87-3.92 (m; 2H, -NH (CH₂) -C≡CH)
4.48-4.53 (m; 1H, -CHCH₂CH₂SH)
8.08-8.12 (m; 2H, 2x -NH-)
8.22 (t; 1H, -NH-)
8.32 (t, 1H, -NH-)

b) Technetium-99m complex [11b] of the N-decazioylhomo cysteinylglycylglycylpropargylamide

A solution of 0.5 mg (1.1 µmol) N-decanoyl homocystei nylglycylglycylpropargylamid [11a] in 300 µl DMSO mixed with 30 ul 1 N sodium hydroxide solution. Then sets 50 µl of a 0.15 molar trisodium citrate dihydrate Solution and 0.4-0.9 mCi of a pertechnetate solution a Mo-99 / Tc-99m generator. The reaction mixture is portionwise with 10 ul 0.2 molar Tin (II) chloride dihydrate solution added, 10 minutes  incubated 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 (Na₂HPO₄; 0.01 M; pH 2.0); Eluent B: acetonitrile / phosphate buffer (Na₂HPO₄; 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 comes with a solution 1.7 g (10.0 mmol) of glycylglycylpropargylamide in DMF transferred. After adding 1 ml of triethylamine, 3 Heated under reflux for hours. Then under concentrated pressure, the residue in CH₂Cl₂ taken up and washed with dilute HCl. The orga African phases are washed neutral with water and dried over Mg₂SO₄. After narrowing the solution by means of reduced pressure from methanol recrystallized and optionally by column chromatography topography with CH₂Cl₂ / MeOH (9: 1) cleaned.

Yield: 60% of the title compound [12a]
C₁₇H₂₈N₄O₄S (MG: 384.50)
1 H-NMR (d₆-DMSO):
δ = 0.86 (t; 3H, - (CH₂) ₂CH₃)
1.20-1.31 (m; 12H, - (CH₂) ₂CH₃)
1.47-1.54 (m; 2H, -CH₂-CH₂CONH-)
2.10-2.19 (m; 2H, -CH₂-CH₂CONH-)
2.56-2.68 (m; 2H, -CHCH₂CH₂SH)
3.11 (t; 1H, -C≡CH)
3.36-3.40 (m; 2H, -CHCH₂CH₂SH)
3.51 (s; 1H, -SH)
3.67-3.76 (m; 4H, 2x -NH (CH₂) CO-)
3.85-3.89 (m; 2H, -NH (CH₂) -C≡CH)
4.47-4.54 (m; 1H, -CHCH₂CH₂SH)
8.03 - 8.10 (m; 2H, 2x -NH-)
8.20 (t; 1H, -NH-)
8.29 (t; 1H, -NH-)

b) Technietium-99m complex [12b] of the N-hexazioylhomo cysteinylglycylglycylpropargylamide

A solution of 0.5 mg (1.3 µmol) N-hexanoyl homocystei nylglycylglycylpropargylamid [12a] in 300 µl DMSO mixed with 30 ul 1 N sodium hydroxide solution. Then sets 50 µl of a 0.15 molar trisodium citrate dihydrate Solution and 0.4-0.9 mCi of a pertechnetate solution a Mo-99 / Tc-99m generator. The reaction mixture is portionwise with 10 ul 0.2 molar Tin (II) chloride dihydrate solution added, 10 minutes incubated 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 (Na₂HPO₄; 0.01 M; pH 2.0); Eluent B: acetonitrile / phosphate buffer (Na₂HPO₄; 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 13 a) 3,17β-Dihydroxy-17α- [5- (2-benzoylthioacetylglycyl glycyl) amino-pent-1- (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-benzoylthioacetylglycylglycylpro pargylamide [1a], 10.0 mg (0.044 mmol) benzyltriethylam monium chloride, 10.6 mg (9.2 µmol) tetrakis (triphenyl phosphine) palladium (0) and 8.15 mg (0.043 mmol) Copper (I) iodide in 4 ml of toluene is left at room for 72 hours temperature stirred. Then you add Water, extracted twice with toluene and dried over Mg₂SO₄. After concentrating the solvent under ver reduced pressure is taken up in 5 ml THF, with 150 mg (0.6 mmol) pyridinium-para-toluenesulfonic acid in 5 ml Ethanol was added and the mixture was heated under reflux for 3 h. The solvent is then reduced Concentrated pressure and the residue by column chromatography graph with CH₂Cl₂ / MeOH (8: 1) cleaned.

Yield: 41% of the title compound [13a]
C₃₆H₄₁N₃O₆S (MG: 643.80)
1 H-NMR (d₆-DMSO):
δ = 0.83 (s; 3H, -CH3 steroidal)
1.15-2.30 (m; 13H, steroidal)
2.65-2.74 (m; 2H, -CH₂- steroidal)
3.72 (d; 2H, -NH (CH₂) CO-)
3.81 (d; 2H, -NH (CH₂) CO-)
3.90 (s; 2H, -SCH₂CO-)
4.02 (d; 2H, -NH (CH₂) -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-99m complex [13b) of 3,17β-dihydroxy 17α- [5- (2-benzoylthioacetylglycylglycyl) amino pent-1- (E) -en-3-in] -1,3,5-estratriene

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

The analysis of the marking is done by 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 (Na₂HPO₄; 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 14 a) 17β-Hydroxy-17α- [5- (2-benzoylthioacetylglycyl glycyl) amino-pent-1- (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) S-Ben zoylthioacetylglycylglycylpropargylamide [1a], 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) of copper (I) iodide in 4 ml of toluene Stirred for 72 hours at room temperature. Subsequently water is added and the mixture is extracted twice with toluene and dries over Mg₂SO₄. After narrowing the Solvent under reduced pressure is in 5 ml THF taken up with 150 mg (0.6 mmol) of pyridinium para toluenesulfonic acid in 5 ml of ethanol and 3 hours heated under reflux. Then the Lö removed under reduced pressure and the Residue by column chromatography with CH₂Cl₂ / MeOH (8: 1) cleaned.

Yield: 39% of the title compound [14a]
C₃₆H₄₃N₃O₆S (MG: 645.80)
1 H-NMR (d₆-DMSO):
δ = 0.95 (s; 3H, -CH₃ steroidal)
0.82-2.50 (m; 20H, steroidal)
3.68 (d; 2H, -NH (CH₂) CO-)
3.80 (d; 2H, -NH (CH₂) CO-)
3.87 (s; 2H, -SCH₂CO-)
3.94-3.98 (m; 2H, -NH (CH₂) -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 17β-hydroxy-17α- [5- (2-benzoylthioacetylglycylglycyl) amino-pent-1- (E, Z) -en-3-in] -4-estren-3-one

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 added and 15 minutes at room temperature incubated temperature. Then you put 50 ul one 0.15 molar trisodium citrate dihydrate solution and 2.5 µl a 0.2 molar tin (II) chloride dihydrate solution. The reaction mixture is with a pertechnetate solution (0.4-0.9 mCi) from a Mo-99 / Tc-99m generator ver sets, incubated at room temperature for 10 minutes and then filtered (0.2 µm filter).

The analysis of the marking is done by 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 (Na₂HPO₄; 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 sarcosylglycylpropar gylamid in 30 ml of anhydrous THF is slowly added a solution of 5.3 g (8.4 mmol) of S-acetylmercaptoes acetic acid N-hydroxysuccinimide ester in anhydrous THF transferred. After 4 hours of stirring at room temperature heated to 40 ° C for 30 minutes. After cooling down the solvent was concentrated under reduced pressure and the remaining residue by column chromatography phie over silica gel [CH₂Cl₂ / CH₃OH (9: 1)] cleaned. The Product crystallizes from ethanol.

Yield: 88% of the title compound [15a]
C₁₂H₁₇N₃O₄S (MG: 299.35)
1 H-NMR (d₆-DMSO):
δ = 2.34 (s; 3H, -COCH₃)
2.81 (s; 3H, -N-CH₃)
3.07 (t; 1H, -C≡CH)
3.70 (d; 2H, -NH (CH₂) CO-)
3.82 (s; 2H, -NH (CH₂) CO-)
3.86-3.90 (m; 2H, -NH (CH₂) -C≡CH)
3.93 (s; 2H, -SCH₂CO-)
8.13 (t; 1H, -NH-)
8.22 (t; 1H, -NH-)

b) Technetium-99m complex [15b) of S-acetyl-thioace tylsarcosylglycylpropargylamide

A solution of 0.5 mg (1.67 µmol) S-acetylthioace tylsarcosylglycylpropargylamid [15a] in 50 µl 0.1 N Sodium hydroxide solution is mixed with 250 µl phosphate buffer (Na₂HPO₄,  0.5 mol / l, pH 8.5). Then you set 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 too. The reaction mixture is with a Pertech netate solution (0.4-0.9 mCi) from a Mo-99 / Tc-99m gene rator added, 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 (Na₂HPO₄; 0.01 M; pH 2.0); Eluent B: acetonitrile / phosphate buffer (Na₂HPO₄; 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) succiziylsarcosylglycylpropargyl amide [16a]

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

Yield: 90% of the title compound [16a]
C₁₄H₁₉N₃O₆S (MG: 357.39)
1 H-NMR (d₆-DMSO):
δ = 2.34 (s; 3H, -COCH₃)
2.78 (s; 3H, -N-CH₃)
2.70-3.04 (m; 2H, - (CH₂) COOH)
3.07 (t; 1H, -C≡CH)
3.68-3.73 (m; 4H, 2x -NH (CH₂) CO-)
3.85-3.89 (m; 2H, -NH (CH₂) -C≡CH)
4.33-4.38 (m; 1H, -S-CH (CH₂COOH) -CH₂-)
8.12 (t; 1H, -NH-)
8.23 (t; 1H, -NH-)
12.68 (broad; 1H, -COOH)

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

A solution of 0.5 mg (1.4 µmol) of 2- (S-acetyl thio) succinylsarcosylglycylpropargylamide [16a] in 50 µl 0.1 N sodium hydroxide solution with 250 ul phosphate buffer (Na₂HPO₄, 0.5 mol / l, pH 8.5) diluted. Subsequently put 50 µl of a 0.15 molar trisodium citrate di hydrate solution and 2.5 µl of a 0.2 molar tin (II) - chloride dihydrate solution. The reaction mixture is with a pertechnetate solution (0.4-0.9 mCi) from one Mo-99 / Tc-99m generator offset, 10 minutes at Incubated 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 (Na₂HPO₄; 0.01 M; pH 2.0); Eluent B: acetonitrile / phosphate buffer (Na₂HPO₄; 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 (2- (S-acetylthio) succinylsarcosylglycylpropar gylamid-4-yl) -Cys-Ser-Cys-Ser-Ser-Leu-Met-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 28 mg of dicyclohexylcarbodiimide in anhydrous DMF are added. The reaction mixture is stirred at -5 ° C for 2 hours, then at room temperature for 2 hours and finally cooled to -15 ° C. The 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 neutralizing the buffered eluate, 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
best. 2831.6 (FAB-MS)

b) Technetium-99m complex [17b] des (2- (S-acetyl thio) 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 (Na₂HPO₄₁ 0.5 mol / l, pH 8.5) ver thins. Then 50 µl of a 0.15 molar is placed Trisodium citrate dihydrate solution and 2.5 µl of a 0.2 molar tin (II) chloride dihydrate solution. The Reacti mixture is mixed with a pertechnetate solution (0.4-0.9 mCi) from a Mo-99 / Tc-99m generator, 10 Incubated for 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 (Na₂HPO₄; 0.01 M; pH 2.0); Eluent B: acetonitrile / phosphate buffer (Na₂HPO₄; 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-Propar gylamide [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 1 -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 existing protective groups 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 kept under protective gas (Ar).
MG: calc. 865.0
best. 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-Propargylamid [18a] in 300 µl phosphate buffer (Na₂HPO₄, 0.5 mol / l, pH 8.5) with 50 ul 0.15 molar trisodium citrate dihydrate solution and 2.5 µl a 0.2 molar tin (II) chloride dihydrate solution transferred. The reaction mixture is with a Pertech netate solution (0.4-0.9 mCi) from a Mo-99 / Tc-99m gene rator added, 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 (Na₂HPO₄; 0.01 M; pH 2.0); Eluent B: acetonitrile / phosphate buffer (Na₂HPO₄; 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 from WBHL rabbits

The labeling of the N- (3-hexadecylaminocarbonyl-2- acetylthiopropionyl) glycylglycylpropargylamids (her made according to Example 9a) as in Example 9b described.

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 autoradiography of the aorta and Sudan III staining were carried out to show the atherosclerotic plaques ( Fig. 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 (19)

1. Compounds of the general formula (I), characterized in that (A¹), (A²) and (A³) are the same or different and 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
R³ and R⁵ are the same or different, represent a hydrogen atom, a methyl or an ethyl group and
R⁴ represents a hydrogen atom, a branched or unbranched alkyl group with 1-4 carbon atoms, the C atoms of which may be with amino groups, N (RaR ^b ) groups (where Ra and R ^{b are the} same or different and for branched or unbranched alkyl or acyl radicals with 1-20 carbon atoms, the C atoms of which are maximally substituted with a hydroxyl, a carboxy or an amino group), hydroxyl groups, thiol groups, halogen atoms, carboxy groups, alkoxycarbonyl groups with 1-20 carbon atoms, acyloxy groups with 1- 12 carbon atoms, amino carbonyl groups, sulfonyl groups, aminosulfonyl groups or phosphoric acid residues are substituted,
R² and R⁹ are the same or different and have the same meaning as R⁴,
k, 1 and in 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, a halogen atom, a hydroxy group, an amino group, an N (RaR ^b ) group (where Ra and R ^{b are the} same or different and for branched or unbranched alkyl or acyl radicals with 1-20 carbon atoms stand, whose C atoms are optionally substituted with a hydroxyl, a carboxy or an amino group), a hydrazine group, a hydrazide group, a cholesteryl oxycarbonylmethylaminocarbonyl group, a cholesteryloxycarbonyl group, a cholesteryloxy carbonylmethyloxycarbonyl group, another steroid or a derivative of an ethinyl or Ethenyl steroids,
a substituent of the formula Z¹-Y¹- (CH₂) _i -Q-CO
Q is an -NH- or -O-,
Z¹ has the same meaning as Z,
Y¹ has the same meaning as Y and
i have the same meaning as m,
a substituent of formula V-U-Q¹-wherein
Q¹ is -NH-, -CO- or -O-,
U is a bond, a group of the formula - (OCH₂CO) _h with h = 1-3 or a suitable linker for coupling to bio or macromolecules and
V is a hydrogen atom, a hydroxyl group, an N (RaR ^b ) group (where Ra and R ^{b 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 with a Hydroxy, a carboxy or an amino group are substituted), 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 hydroxy, carboxy, alkoxy, amino or substituted amido groups with 1-20 carbon atoms in the alkyl and / or aryl radical means,
Z represents a hydrogen atom, a halogen atom, a carboxy group, a hydroxy group, an alkoxy carbonyl group having 1-20 carbon atoms, an acyloxy group having 1-20 carbon atoms, an alkoxy group having 1-20 carbon atoms, a cholesteryloxycarbonylmethylaminocarbonyl group, a cholesteryloxycarbonyl group, a cholesteryloxycarbonylmethyloxycarbonyl group, another Steroid or a derivative of an ethine or ethene steroid,
a substituent of formula V-U-Q¹-wherein
Q¹ is -NH-, -CO- or -O-,
U is a bond, a group of the formula - (OCH₂C = O) h- with h = 1-3 or a suitable linker for coupling to bio or macromolecules and
V is a hydrogen atom, a hydroxyl group, an N (RaR ^b ) group (where Ra 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 may be with a hydroxy -, a carboxy or an amino group), a bio- or macromolecule, means,
M represents an element of atomic number 43 or 75,
R¹ has the same meaning as R⁴,
R⁶, R⁷ and R⁸ are the same or different and, for a hydrogen atom, an alkyl group having 1-4 carbon atoms, the C atoms of which are optionally substituted by hydroxy, carboxy or amino groups or for a group of the formula -CH₂-X, in the X has the meaning given above,
and their water-soluble salts. 2. Compounds according to claim 1, characterized in net that Z is a hydrogen atom, a steroid, a Is ethinyl steroid or ethenyl steroid. 3. Compounds according to claim 2, characterized net that X is a hydrogen atom, a halogen atom, a carboxy group, an amino group or one Amido group with 1-20 carbon atoms in the alkyl and / or aryl radical. 4. Compounds according to claim 3, characterized net that Y is an ethinylidene group, R¹, R³, R⁶, R⁷ and R⁸ are hydrogen atoms and k, l and m the number 0 mean.   5. Compounds according to claim 1, characterized net that Z is a hydrogen atom and X is a Cho lesteryloxycarbonylmethylaminocarbonyl group, a Cholesteryloxycarbonyl group, a cholesteryloxy carbonylmethyloxycarbonyl group, another Steroid or a derivative of an ethynyl or Ethenylsteroids is. 6. Compounds according to claim 5, characterized in net that Y is an ethinylidene group, R¹, R³, R⁶, R⁷ and R⁸ are hydrogen atoms and k, 1, and m the numbers 0 or 1 mean. 7. Compounds according to claim 1, characterized in that Z is a hydrogen atom and X is a hydrogen atom, a carboxy group or a substituent of the formula VU-Q¹-wherein
Q¹ is -NH- -CO- or -O-,
U is a bond, a group of the formula - (OCH₂CO) _h - with h = 1-3 or a suitable linker for coupling to bio or macromolecules and
V is a hydrogen atom, a hydroxyl group, an N (RaR ^b ) group (where Ra and R ^{b 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 with a Hydroxy, a carboxy or an amino group are substituted), a bio or macromolecule means. 8. Compounds according to claim 7, characterized in net that Y is an ethinylidene group, R¹, R³, R⁶, R⁷ and R⁸ are hydrogen atoms and k, l and m the numbers mean 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 amino carbonyl group, a sulfonyl group, an amino sulfonyl group, a phosphoric acid residue, a carboxymethylaminocarbonyl group, a p-amino phenyl group, a p-hydroxyphenyl group, a halogen residue, a hydroxy group, an amino group, an N (RaR ^b ) group (where Ra and R ^{b are the} same or different 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 or a hydrazide group . 10. Compounds according to claim 9, characterized in net that Y is an ethinylidene group, R¹, R³, R⁶, R⁷ and R⁸ are hydrogen atoms and k, l and m the numbers mean 0, 1 or 2.   11. Compounds of the general formula (II), characterized in that A¹, A², A³, R¹, R⁶, R⁷, R⁸, Y and Z have the meaning given in claim 1 and in which 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 hydroxy acetyl group or another suitable sulfur protecting group. 12. Conjugates containing compounds of the general Formula I and / or II and selectively getting sick substances enriching the tissues, whereby between there is a covalent bond between them and this sub in the case of amino groups punch like peptides, proteins and antibodies or their fragments, amidic or in the case of Substances containing hydroxyl groups, such as fatty alcohol alcohols, ester-like or in the case of aldehyde group Pen-containing substances are imidic.   13. Conjugates according to claim 12, characterized in net that accumulate in the diseased tissue the substances peptides, especially endotheline, Partial sequences of endothelins, endothelin ana loga, endothelin derivatives or endothelin antago nesting mean. 14. Conjugates according to claim 12, characterized in that the peptides the sequences or parts thereof 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 general formula (II) wherein
A¹, A², A³, R¹, R⁶, R⁷, R⁸, 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
  A¹, A², A³, R¹, R⁶, R⁷, R⁸, Y and Z have the meaning given in Claim 1 and Hal is a halogen,
  with a compound of the general formula T-SM⁺worin M⁺ is an alkali metal cation and T has the meaning given in claim 11,
  or
• b) a compound of the general formula HN (R⁶) -A²-N (R⁷) -A³-N (R⁸) -Y-Zworin A², A³, R⁶, R⁷ and R⁸ have the meaning given in claim 1 and
  with a compound of the general formula T-S-A¹-Wworin A¹ and T have the meaning given in claim 11 and W has the meaning of a leaving group which enables A¹ to react with free amino groups.

17. Kit for the production of radiopharmaceuticals, consisting from a compound of the general formula (II) according to claim 11, a reducing agent and counter also an auxiliary ligand, which is in dry State or in solution, a use instruction with a reaction instruction for implementation tion of the described connections with Techne tium-99m or rhenium in the form of a pertechnetate solution or perrhenate solution. 18. Radiopharmaceutical composition for not invasive in vivo imaging of receptors and tissue containing receptors and / or atherosclerosis table plaques, characterized in that they a compound according to claim 1, optionally with the additives common in galenics and that the compound of claim 1 in one Kit according to claim 17 with technetium-99m or Rhenium in the form of a pertechnetate solution or Perrhenate solution is prepared. 19. Radiopharmaceutical composition according to claim 18, characterized in that it is a verb dung according to claim 1 in the form of liposomes and that the compound of claim 1 in one Kit according to claim 17 with technetium-99m or Rhenium in the form of a pertechnetate solution or Perrhenate solution is prepared.