DK171574B1 - Macrocyclic compounds, diagnostic agent containing the compounds and use of the compounds - Google Patents

Abstract

Compounds of the general formula I <IMAGE> in which Y is a nitrogen or phosphorus atom, A<1> and A<2> are identical or different and are each a straight-chain or branched alkylene group having 2 to 6 carbon atoms, U<1>, U<2>, U<3>, U<4> are identical or different and are each a direct bond or a straight-chain or branched alkylene group having 1 to 6 carbon atoms, D<1>, D<2>, D<3>, D<4> are identical or different and are each an oxygen or sulphur atom, an alkylene group having 1 to 6 carbon atoms or a group N-R<7> where R<7> has the meaning of a hydrogen atom, a straight-chain or branched alkylene chain having 1 to 4 carbon atoms, which carries a COOR<1> group at the end, in which R<1> represents a hydrogen atom or a metal ion equivalent, D<5> has the meaning given for D<1>, D<2>, D<3> and D<4> and the group <IMAGE> where R<5> has the meaning of a hydrogen atom or of a straight-chain or branched, saturated or unsaturated C1-C20-alkylene group which may contain imino, phenyleneoxy, phenyleneimino, amide, ester groups, oxygen, sulphur and/or nitrogen atoms and may be substituted by hydroxyl, mercapto, imino and/or amino groups and has either a functional group or bound via this group a macromolecule B at the end, s and t are integers from 0 to 5, R<2> is hydrogen, a linear or branched, saturated or unsaturated, alkyl, acyl or acylalkyl group of 1 to 16 carbon atoms which is unsubstituted or substitute by one or more hydroxyl or lower alkoxy groups, or is -CH2-X-V where X has the meaning of carbonyl, of a straight-chain or branched alkylene group having 0 to 10 carbon atoms, which is unsubstituted or substituted by one or more hydroxyl or lower alkoxy groups, or of a straight &tilde& chain or branched alkylene group having 2 to 23 carbon atoms which is interrupted by oxygen atoms, V has the meaning <IMAGE> or -COOR<6> in which R<3> and R<4>, independently of one another, are hydrogen, a linear o branched alkyl group having 1 to 16 carbon atoms which is unsubstituted or substituted by one or more hydroxyl or lower alkoxy groups, or R<3> and R<4> together with the nitrogen atom represent a saturated 5- or 6-membered ring which may contain a further hetero atom and R<6> represents hydrogen or a saturated, unsaturated, straight-chain or branched or cyclic hydrocarbon radical having up to 16 carbon atoms or represents an aryl or aralkyl group, or R<2> or R<3> denotes a second macrocycle of the formula I' <IMAGE> which is bound via an alkylene chain (K) containing 2 to 20 carbon atoms, which, if desired carries carbonyl groups at the ends and, if desired, is interrupted by one or more oxygen atoms or R<1>-carboxymethylimino groups or substituted by one or more hydroxyl, lower alkoxy or carboxy-lower alkyl groups, which macrocycle can have a structure which is different from that of the parent structure, or R<2> denotes B or CH2-COB, with the proviso that if Rz represents B or CH2-COB, R<5> denotes a hydrogen atom, that at least two COORi groups are present in the molecule and that two hetero atoms of the macrocycle are each bound to at least two carbon atoms via an alkylene group and functional groups present in the molecule are, if desired conjugated with macromolecules and, if desired free carboxyl groups have been converted with organic or inorganic bases or amino acids and basic groups with inorganic or organic acids to the salts.

Description

DK 171574 B1

The invention relates to novel macrocyclic 1,4,7,10-tetrazacyclododecan derivatives of the general formula I shown below, a diagnostic agent containing one or more such compounds, and the use of the compounds as complexing agents or in the preparation of diagnostics or therapeutic agents. agents.

Since the early 1950s, metal complexes have been used as contrast agents in radiology. However, the compounds used at that time were toxic to the extent that human use was not considered. Therefore, it was very surprising that certain complex salts proved to be sufficiently compatible to allow routine application to humans for diagnostic purposes.

As the first example of this class of substances, the dimeglumine salt of Gd-DTPA (Gadolinium-III complex of diethylenetriamine pentaacetic acid) disclosed in the European Patent Application No. 71564 has so far proved to be a promising contrast agent for nuclear spin tomography in clinical studies of over 1000 patients. The main area of application 20 lies in diseases of the central nervous system.

A major reason for this good compatibility of Gd-DTPA in clinical use lies in the high efficacy of nuclear sphincter tomography, especially in many brain tumors. Due to the good effect, Gd-DTPA can be dosed at 0.1 25 mmol / kg body weight, many times lower than, for example, X-ray contrast agents, in many X-ray examinations.

As a further example of the complex salts, the meglumine salt of Gd-DOTA (Gadolinium-III complex of 1,4,7,10-tetraazacycloododecan tetraacetic acid) disclosed in German Patent Application No. 34 01 052 has been shown to be promising for diagnostic purposes. purpose.

However, it is desirable to be able to use chelates at a higher dose as well. This is particularly the case for the detection of certain diseases outside the central nervous system by nuclear spin tomography (NMR diagnostics), but especially by the use of chelates as X-ray contrast agents.

Chelates, in comparison with iodinated X-ray contrast-5, offer a number of advantages: a) Radiation absorption in the high energetic area, thus reducing the patient's radiation load and improving the conditions of the energy subtraction method.

b) Avoidance of the "contrast agent reactions" known as 10 unpredictable, in part even life-threatening or deadly "allergy-like" or cardiovascular side effects that characterize the previously used iodinated X-ray contrast agents.

Prerequisites for this are: 15 high concentration of radiation absorbing elements in the solution (X-ray) or strong influence on the NMR signals, a diagnostic suitable for diagnostic purposes, especially firm binding of the metal ions in separable complexes, also under in vivo conditions, good compatibility of the high-concentration, high-dose complex solutions, low allergic potential of all the contrast agent components and high stability and storage capacity of the chemical constituents of the contrast solution.

3 DK 171574 B1

These requirements apply to varying degrees and in different ways, but generally they apply to all uses of the mentioned complexes in in vivo diagnostics.

The compounds of the invention and the solutions prepared therefrom surprisingly satisfy the aforementioned requirements. They have a powerful effect that can be adapted to the current principles of the diagnostic method (X-ray, NMR, ultrasound, nuclear diagnostics).

The compounds of the invention are used: 10 1. For NMR diagnostics in the form of their complexes with the ions of the transition metals having the atomic numbers 21 to 29, 42 and 44.

2. For NMR and X-ray diagnostics in the form of their complexes with the ions of the lanthanide elements with atomic numbers 57 to 70.

3. For the ultrasound diagnostics, both the compounds intended for use in the NMR diagnostics and also those intended for use in the X-ray diagnostics are suitable.

4. For the radio diagnosis and radiotherapy in the form of their 20 complexes with the radioisotopes of the elements with atomic numbers 27, 29, 31, 32, 38, 39, 43, 49, 62, 64, 70 or 77.

Even without special measures, their pharmacokinetics allow an improvement in the diagnosis of numerous diseases. The complexes are again excreted, mostly unchanged and rapidly, so that despite high doses no harmful side effects that can be traced back to the metal can be detected, especially using relatively toxic metal ions as principle of action.

4 DK 171574 B1

The practical use of the novel complexes and complexing agents of the invention is also facilitated by their adequate, even very good, chemical stability.

A further significant advantage of the present complexes 5 and complexing agents is their exceptional chemical versatility.

In addition to the central atom, it is also possible, by appropriate choice of various substituents and / or salt formers, to adapt the properties to the requirements set for effect, pharmacokinetics, compatibility, manageability, etc. In this way, one can be obtained in the diagnosis and therapy. particularly desired specificity of the compounds for the structures of the organism, for certain biochemical substances, for metabolic processes and for the conditions of the tissues or body fluids, in particular by binding to biological substances or to substances which exhibit an interaction with biological systems. Such substances suitable for binding may be low molecular weight (e.g. glucose, amino acids, fatty acids, bile acids, porphyrins) or high molecular weight (polysaccharides, proteins, antibodies, etc.) or may also be body-promoting structures which, however, distribute to a particular way in the body, or which reacts with the body's components. The use of such principles will be all the more possible, the more sensitive the detection method for a diagnostic or the more effective e.g. a radiolabeled complex is in the therapy.

The compounds of the invention may also be used for radiotherapy in the form of their complexes with radioisotopes which

* 1 Q H

for example, Ir. Furthermore, the complexing agents of the invention are suitable as such or in the form of weak complexes with preferably the body's own ions (Ca 2+, Mg 2+, 30 Zn 2+, Fe 2+ // 2 +) for the therapy of heavy metal poisoning or certain storage disorders.

The macrocyclic compounds, i.e. The 1,4,7,10-tetraazacyclo-dodecane derivatives of the invention are characterized in that they have the general formula I

Wherein each of the groups R 1 is independently hydrogen or a metal ion equivalent, R is a straight or branched, saturated or unsaturated hydrocarbon group having 1 to 16 carbon atoms, optionally substituted with 1-5 hydroxy or C 1 -C 4 alkoxy groups, or R 3

R 1 is -CH, -X-N

X is a carbonyl or a straight chain or branched alkylene group having 1 to 10 carbon atoms optionally substituted with 1-5 hydroxy or C 1 -C 4 alkoxy groups, or X is a straight chain or branched alkylene group having 5 to 23 carbon atoms, interrupted by oxygen atoms and wherein R 3 and R 4 are independently hydrogen, a straight or branched alkyl group having 1 to 16 carbon atoms, optionally substituted with 1-5 hydroxy or C 1 -C 2 alkoxy groups, or R 3 and R 4 together with the nitrogen atom are a pyrrolidine, 20 piperidine, morpholine or piperazine ring, or R or RJ is one via an alkylene chain of 2 to 20 carbon atoms which optionally carries terminal carbonyl groups and is optionally interrupted by 1-4 oxygen atoms or which is substituted by 1-5 hydroxy, C 1 -C 2 -alkoxy or carboxy-C 1 -C 2 alkylene groups, second linked macrocycle of formula I 'DK 171574 B1 i, \ nf.

R1OOC_ / N_ / \ __ COOR

or represents B or CH2-COB, where B represents a macromolecule or biomolecule that accumulates to a great extent in the organ or organ moiety being examined or in the tumor, and salts thereof with organic or inorganic bases or amino acids or with inorganic or organic acids.

Compounds of general formula I, wherein R 1 is hydrogen, are referred to as complexing agents and when at least two of the substituents R 1 have a metal ion equivalent, the compounds are referred to as metal complexes.

If not all acidic hydrogen atoms are substituted with the central ion, one or more or all of the remaining hydrogen atoms may be replaced by cations of inorganic and / or organic bases or amino acids. Suitable inorganic cations are, for example, the lithium ion, the potassium ion, the calcium ion and especially the sodium ion. Suitable cations of organic bases are, among others, those of primary, secondary or tertiary amines, such as e.g. ethanolamine, diethanolamine, morpholine, glucamine, N, N-dimethylglucamine and especially N-methylglucamine. Suitable cations of 20 amino acids are, for example, cations of lysine, arginine and ornithine.

The complex compounds can also be linked to macromolecules, which are known to be stored in particular in the organs or organ parts to be examined. Such macromolecules are, for example, hormones, dextrans, polysaccharides, polychelones, hydroxyethyl starch, polyethylene glycols, desferrioxamines, bleomycins, insulin, prostaglandins, steroid hormones, amino sugars, amino acids, peptides such as polylysine, proteins (such as immunoglobulins) and proteins (such as immunoglobulins). or lipids (also in the form of liposomes). In particular, mention should be made of conjugates with albumins such as human serum albumin, antibodies such as e.g. monoclonal antibodies specific for tumor-associated antigens or antimyosin. Suitable synthetic polymers, such as polyethylene imines, may also be substituted for the biomolecules. The diagnostic agents thus obtained are suitable, for example, for use in tumor and infarction diagnostics. In particular, monoclonal antibodies for the conjugation are those which are predominantly directed against cell membrane-associated antigens. As such are suitable, e.g. for tumor detection, monoclonal antibodies or their fragments (F (ab) 2) / such as e.g. is directed to the 20 carcinoembryonic antigen (CEA), human choriogonadotropin (β-hCG), or other tumor-associated antigens such as glycoproteins. Suitable include antimyosin as well as anti-insulin and anti-fibrin antibodies.

For example, liver tests or tumor diagnostics are suitable for example conjugates or inclusion compounds with liposomes (used, for example, as unilamellar or multilamellar phosphatidylcholine-cholesterol vesicles).

The preparation of the macrocyclic compounds of general formula I is carried out by substituting in compounds of general formula I, which instead of the COOR1 group, the group COOZ, where Z is a carboxy protecting group, without, however, including a macromolecule B or the group CH2-COB, the protecting groups Z decompose, after which the acids thus obtained (R1 of the general formula I represent hydrogen), if desired, are reacted with at least one metal oxide in a manner known per se; metal salt of an element having atomic numbers 21-29, 31, 32, 38, 39, 42-44, 49, 57-70 or 77, if desired, acidic hydrogen atoms present by means of inorganic and / or organic bases or amino acids and present basic groups by means of inorganic or organic acids are then transferred into physiologically acceptable salts, or b) reacted in a manner known per se with at least one metal oxide or metal salt of an element having atomic numbers 21-29, 31 , 32, 38, 39, 42-44, 57-70 or 77, after which the metal complexes obtained are then bonded in a manner known per se to a macromolecule via the functional groups contained in the molecule or the CO group contained in R2, wherein the 15 acidic hydrogen atoms present by means of inorganic and / or organic bases or amino acids and basic groups present by means of inorganic or organic acids, if desired, are transferred to physiologically acceptable salts or by binding in a manner known per se. macromolecule by means of the functional groups contained in the molecule, or to R 2 or to the CO group contained in R 2, and then react in a manner known per se with at least one metal oxide or metal salt of an element having atomic numbers 25 to 21 , 31, 32, 38, 39, 42-44, 49, 57-70 or 77, and if desired, the acidic hydrogen atoms present are transferred by means of inorganic and / or organic bases or amino acids and the t oxygen-containing basic groups by means of inorganic or organic acids for physiologically acceptable salts.

As carboxyl protecting groups Z are lower alkyl, aryl and aralkyl groups, for example methyl, ethyl, propyl, butyl, phenyl, benzyl, diphenylmethyl, triphenylmethyl and bis ( p-nitrophenyl) methyl group and the trialkylsylyl group.

The cleavage of the protecting groups Z occurs in a manner known per se, for example by hydrolysis, alkaline saponification of the esters, preferably with alkali in aqueous-alcoholic solution at temperatures of 0 to 50 ° C or, in the case of e.g. tert-butyl esters, using trifluoroacetic acid.

The products are prepared by cycling two reactants. The cyclic compounds thus obtained are then reacted, optionally after cleavage of the protecting groups, with haloalkanes, esters, acids, ketones, acyl halides or anhydrides, to introduce the substituent R2.

Cyclization is carried out according to methods known in the literature (e.g., Org. Synth. 58 .. 86 (1978), Macrocyclic Polyether Syntheses, Springer Verlag Berlin, Heidelberg, New York 1982, Coord. Chem. Rev. 3. / 3 (1968), Ann. Chem. 1976, 916): one of the two reactants carries at the chain end two leaving groups, the other two nucleophiles displacing these leaving groups. For example, the reaction of terminal, optionally heteroatom-containing dibromo, dimesyloxy, ditosyloxy or dialkoxycarbonylalkylene compounds with terminal diazaalkylene compounds, which optionally contains additional heteroatoms, one of the two reactants being R 5

Nitrogen atoms present may have been protected, e.g. as tosylates, and are released prior to the subsequent alkylation reaction according to methods known from the literature.

If diesters are used in the cyclization reaction, the thus obtained diketo compounds must be reduced according to methods known to those skilled in the art, e.g. with diborane.

10 DK 171574 B1

The subsequent alkylation is carried out with halogen ketones, esters, acids or alkanes which may be substituted by one or more hydroxy or lower alkoxy groups and optionally contain oxygen atom (s) in the chain. Furthermore, the alkyl group may also contain terminal amino groups.

When the alkylation is carried out with a dihalogenated alkane, compounds of the general formula I are obtained with two macrocyclic rings bonded to each other via a hydrocarbon bridge.

Additional methods known from the literature for the synthesis of compounds having more than one ring are e.g. reacting an amine with a carbonyl compound (e.g., acid chloride, mixed anhydride, activated ester, aldehyde); two amine-substituted rings with a dicarbonyl compound (e.g.

Oxalyl chloride, glutardialdehyde); two rings, each having a nucleophilic group, and an alkylene compound having two leaving groups; in the case of terminal acetyl groups, oxidative coupling may be cited (Cadiot, Chodkiewicz in Viehe "Acetylenes", 597-647, Marcel Dekker, New York, 1969). The 20 chains linking the rings can then be modified by sequential reactions (e.g., hydration).

In the alkylation with haloacetic acid, an intermediate of R 2 = CH 2 X-COOH is obtained which is converted to the monoamide via the mixed anhydride with chloroformic acid ester or by dicyclohexylcarbodiimide and reaction with a primary or secondary amine of the general formula R

HN

30 \.

R4.

11 DK 171574 B1

Suitable amines include, for example: dimethylamine, diethylamine, di-n-propylamine, diisopropylamine, di-n-butylamine, diisobutylamine, di-butylamine, N-methyl-n-propylamine, dioctylamine, dicyclohexylamine, N-ethylcycloamine. Hexylamine, diisopropenylamine, benzylamine, aniline, 4-methoxy-aniline, 4-dimethylaminoaniline, 3,5-dimethoxyaniline, morpholine, pyrrolidine, piperidine, N-methylpiperazine, N-ethylpiperazine, N- (2-hydroxyethyl) -piperazine, N- (hydroxymethyl) -piperazine, piperazinoacetic acid isopropylamide, N- (piperazinomethylcarbonyl) -morpholine, N- (piperazinomethylcarbonyl) -pyrrolidine, 2- (2-hydroxymethyl) piperidine, 4- (2-hydroxyethyl) -piperidine, 2-hydroxymethylpiperidine, 4-hydroxymethylpiperidine, 2-hydroxymethylpyrrolidine, 3-hydroxypiperidine, 4-hydroxypiperidine, 3-hydroxypyrrolidine, 4-piperidone, 3-pyrroline, piperidin-3-carboxylic acid amide 4-carboxylic acid amide, piperidine-3-carboxylic acid diethylamide, piperidine-4-carboxylic acid methylamide, 2,6-dimethylpiperidine, 2,6-dimethylmorpho lin, N-acetyl-piperazine, N- (2-hydroxy-propionyl) -piperazine, N- (3-hydroxy-propionyl) -piperazine, N- (methoxyacetyl) -piperazine, 4-20 (N-acetyl-N- methylamino) -piperidine, piperidine-4-carboxylic acid (3-oxapentamethylene) -amide, N- (Ν ', N'-dimethyl-carbamoyl) piperazine, pyrazoline, pyrazolidine, imidazoline, oxazolidine, thiazolidine, 2.3- dihydroxypropylamine, N-methyl-2,3-dihydroxypropylamine, 2-hydroxy-1- (hydroxymethyl) ethylamine, N, N-bis- (2-25 hydroxyethyl) amine, N-methyl-2,3,4 , 5,6-pentahydroxyhexylamine, 6-amino-2,2-dimethyl-1,3-dioxepin-5-ol, 2-hydroxyethylamine, 2-amino-1,3-propanediol, diethanolamine, ethanolamine.

The polyhydroxyalkylamines may also advantageously be used in protected form for the reaction: e.g. as O-acyl derivatives or as ketals. This is especially true when these derivatives can be made easier and cheaper than the polyhydroxyalkylamines themselves. A typical example is 2-amino-1- (2,2-dimethyl) -1,3-dioxolan-4-yl) ethanol, the acetonide of 1-amino-2,3,4-trihydroxy-butane, which manufactured in accordance with DE publication no.

35 31 50 917.

The subsequent removal of the protecting groups takes place without problems and can e.g. is carried out by treatment with an acidic ion exchanger in aqueous-ethanolic solution.

DK 171574 Pg 12

The acylation is carried out by a corresponding acyl derivative, in particular with an acyl halide or anhydride. When anhydrides or halides of di- or polycarboxylic acids are used, compounds of the general formula III are obtained in which both macrocyclic rings are bonded to each other via a hydrocarbon bridge.

Examples of conversions of hydroxy or amino groups bound to aromatic or aliphatic groups are the reactions carried out in anhydrous aprotic solvents such as tetrahydrofuran, dimethoxyethane or dimethylsulfoxide in the presence of an acid neutralizing agent.

Such as e.g. sodium hydroxide, sodium hydride or alkali or alkaline earth metal carbonates, such as, for example, sodium, magnesium or potassium calcium carbonate, at temperatures between 0 ° C and the boiling point of the solvent, preferably between 20 ° C and 60 ° C, with a substrate of the general formula IV

W-L-Fu (IV) wherein W is a nucleofug, e.g. Cl, Br, I, Cl 2 Cgl 2 SO 2 or CF 3 SO 3, L represents an aliphatic, aromatic, aryl aliphatic, branched, straight-chain or cyclic hydrocarbon group 25 of up to 20 carbon atoms, and Fu represents the desired terminal functional group.

Examples of compounds of general formula IV include:

Br (CH 2) 2 NH 2, Br <CH 2) 30 H, BrCH 2 COOCH 2, BrCH 2 CO 2 Bu,

Br (CH2 »4CO2C2H5, BrCH2COBr, BrCHjCONH ^, CICHjCOOCjHj, Λ

BrCH2CONHNH2, BrCH2-CH-CH2, CFgSOj (CH2) 3Br, BrC2 CSCH, BrCHjCH-CHj.

13 DK 171574 B1

For example, conversions of carboxyl groups can be carried out according to the carbodiimide method (Fieser, Reagents for Organic Syntheses 10, 142) via a mixed anhydride [Org. Prep. Proc.

Int. 7, 215 (1975)] or via an activated ester (Adv. Org.

5 Chem., Part B, 472).

The compounds of general formula I wherein R 1 represents a hydrogen atom are complexing agents. They can be isolated and purified or transferred without isolation to metal complexes of general formula I, wherein at least two of the substituents R 1 represent 10 a metal ion equivalent.

The preparation of the metal complexes of the invention is carried out in the manner disclosed in DE Publication 34 01 052 and EP 71564, by dissolving or suspending the metal oxide or metal salt (e.g., the nitrate, acetate, carbonate, chloride or sulfate) of the element having atom number 21 -29, 31, 32, 38, 39, 42-44, 49, 57-70 or 77 in water and / or in a lower alcohol (such as methanol, ethanol or isopropanol) and react the solution or suspension with the equivalent amount of it. complexing acid of the general formula I, wherein R 1 represents a hydrogen atom and, if desired, substituting acidic hydrogen atoms present in acid groups with cations from inorganic and / or organic bases or amino acids.

The neutralization is carried out by means of inorganic bases (e.g. hydroxides, carbonates or bicarbonates) of e.g. sodium, potassium or lithium and / or organic bases such as primary, secondary and tertiary amines such as e.g. ethanol-amine, morpholine, glucamine, N-methyl and N, N-dimethylglucamine, as well as basic amino acids, e.g. lysine, arginine and ornithine.

For example, in the preparation of the neutral complex compounds, the acidic complex salts in aqueous solution or suspension can be added to so much of the desired bases as to reach the neutral point. The solution obtained can then be evaporated to dryness under vacuum. It is often advantageous to precipitate the neutral salts formed by the addition of water-miscible solvents, e.g. lower alcohols (methanol, ethanol, isopropanol and others), lower ketones (acetone and 5 others), polar ethers (tetrahydrofuran, dioxane, 1,2-dimethoxyethane and others) to obtain crystallisates which can be readily isolated and purified . It has been found to be particularly advantageous to add the desired base to the reaction mixture already during complex formation, thereby saving a step 10 of the process.

If the acidic complexes contain several free acidic groups, it is often convenient to prepare neutral mixture salts which contain both inorganic and organic cations as counterions.

This may be accomplished, for example, by reacting the complexing acids in aqueous suspension or solution with the oxide or salt of the elements supplying the central ion, and half of the amount of organic base needed for neutralization isolates the resulting complex salt, purifies it if necessary, and then translates it into complete neutralization with the required amount of the organic base. The order of base addition can also be reversed.

Basic groups can be converted into pharmaceutically acceptable salts with inorganic and / or organic acids.

As organic bases, for example, lithium, sodium and potassium hydroxide are used. Suitable organic bases are among other primary, secondary and tertiary amines such as e.g. ethanol-amine, morpholine, glucamine, N-methyl and N, N-dimethylglucamine, as well as basic amino acids such as e.g. lysine, arginine and ornithine.

15 DK 171574 B1

For salt formation with basic groups, inorganic acids, such as e.g. hydrochloric acid, and organic acids such as citric acid, on speech.

For example, the conjugate formation can be carried out via a carboxyl group in the complex compound or via the functional group defined above and which is terminally at the C1-C20 alkylene group in the substituent R5. In the conjugate formation of acids with macromolecules, several acid groups can be attached to them. In this case, 10 more central ions can be attached to a macromolecule.

The coupling to the desired macromolecules also takes place according to methods known per se, which are described, for example, in Rev. Roum. Morphol. Embryol. Physiol., Physiology 1981, 18, 241 and J. Pharm. Sci. , § 79, (1979), for example, by reacting the nucleophilic group in a macromolecule, such as the amino, phenol, sulfhydryl, aldehyde or imidazole group, with an activated derivative of the complexing agent. As activated derivatives, for example, monohydrides, acid chlorides, acid hydrazides, mixed anhydrides (see, e.g., G.E. Krejcarek 20 and K.L. Tucker, Biochem. Biophys. Res. Commun. 1977, 581), activated esters, nitrenes or isothiocyanates are considered.

Conversely, it is also possible to react an activated macromolecule with the complexing acid. For conjugation with 25 proteins, substituents, e.g. with the structure C6H4N2, C6H4NHCOCH2, C6H4NHCS or CgH4OCH2CO.

The conjugation of complexing acids with dextrans and dextrins is also carried out according to known methods, for example by activating polysaccharides with bromocyanine and subsequent reaction with the amino groups of the complexing acids.

16 DK 171574 B1 In case complex compounds containing radioisotopes are used, their preparation can be carried out according to the methods described in "Radiotracers for Medical Applications", Volume 1, CRC-Press, Boca Raton, Florida.

The compositions of the invention fulfill all the prerequisites of being suitable as contrast agents for use in the core spin tomography. Thus, after oral or parenteral use, they are excellent in improving the pronunciation power obtained by the nuclear spin tomography by increasing the signal intensity. Furthermore, they exhibit the high efficiency required to burden the body with as little foreign matter as possible and the good compatibility needed to maintain the non-invasive nature of the studies.

The good water solubility of the compositions of the invention permits the preparation of highly concentrated solutions so that the volume load of the circuit can be kept within acceptable limits and to offset the dilution caused by body fluid. Furthermore, the agents of the invention 20 exhibit not only a high in vitro stability but also a surprisingly high in vivo stability; thus, the release or replacement of the non-covalently bound - in and of itself toxic - ions in the complexes occurs only very slowly during the time required for complete separation of the novel contrast agents.

Generally, the agents of the invention are dosed using as NMR diagnostics in amounts of 0.001 - 5 mmol / kg, preferably 0.005 - 0.5 mmol / kg. Details of the application are discussed, for example, in H.J. Weinmann et al., Am. J.

30 or Roentgenology 142. 619 (1984).

Particularly low doses (below 1 mg / kg) of organ-specific NMR diagnostics can be used, for example, in the detection of tumors and myocardial infarction. Further, complex compound 17 advantageously can be used as "shift reagents" according to the invention.

The agents of the invention, because of their useful radioactive properties and the good stability of the complex compounds contained therein, are also suitable as radio diagnostics. Details of its use and dosage are described, for example, in "Radiotracers for Medical Applications", CRC-Press, Boca Raton, Florida.

An additional imaging method using radioisotopes 10 is the positron emission tomography which uses positron emitting isotopes such as 43Sc, 44Sc, 52Fe, 55Co and 6®Ga.

(Heiss, W.D., Phelps, M.E., Posiiton Emission Tomography of Brain, Springer Verlag Berlin, Heidelberg, New York 1983).

The compounds of the invention may also be used in the radioimmunotherapy. This differs only from the corresponding diagnostics by the amount and nature of the radioactive isotope used. The aim of this is the destruction of tumor cells by energy-rich short-wave radiation with the least possible range. The specificity of the antibodies used is hereby essential, since nonspecificly located antibody conjugates lead to destruction of healthy tissue.

In the following table, the closest compound of the prior art is Gd-DOTA (Formula I, where R 2 = CH 2 COOH and 3 x R 1 = Gd 3+; cf. WO 86/02352, EP 124.766, DE 34 01 052 25 Al, Chemical Abstracts Vol. 97, 1982, 206960z), have been compared to representative compounds of the invention.

It is clear that, by giving up an acetic acid group on a nitrogen atom in the macrocycle, one does not surprisingly experience a deterioration, but on the contrary a very clear improvement in the properties of the complex in question. Thus, the T1 relaxivity values, which are a measure of the signal amplification of the contrast agents used, for example with respect to the compounds of, for example, 5/19, 20 and 21, are clearly higher than the corresponding value for the closest known compound (Gd DOTA).

The osmotic pressure, which is responsible for side effects such as pain, blood vessel damage and cardiovascular disorders, is greatly reduced in the investigated complexes of the invention relative to the clearly hypertonic Gd-DOTA compound.

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The agents of the invention are excellent as X-ray contrast agents, in particular it should be emphasized that, using the agents, no signs of anaphylaxis-like reactions can be seen in biochemical-pharmacological studies known from the iodine-containing contrast agents. They are particularly useful because of their favorable absorption properties in areas of higher tube voltage by digital subtraction techniques.

Generally, the agents of the invention are dosed using as X-ray contrast agents analogous to, for example, meglumine diatrizoate in amounts of 0.1-5 mmol / kg, preferably 0.25-1 mmol / kg.

Details of the use as X-ray contrast agents are e.g. discussed in Barke, Rontgenkontrastmittel, G. Thieme, 15 Leipzig (1970) and P. Thurn, E. Bucheler - "Einfiihrung in die X-ray Diagnostics", G. Thieme, Stuttgart, New York (1977).

Because their acoustic impedance is higher than the impedances of the body fluids and tissues, the agents of the invention are also suitable as contrast agents in the ultrasound diagnostics, especially in the form of suspensions. Generally, they are dosed in amounts of 0.1 to 5 mmol / kg, preferably 0.25 to 1 mmol / kg.

Details of the use as ultrasound diagnostics are described, for example, in T.B. Tyler et al., Ultrasonic Imaging 3.323 (1981), J.I. Haft, "Clinical Echocardiography", Futura, Mount Kisco, New York 1978, and G. Stefan "Echocar Diography" G. Thieme Stuttgart / New York 1981.

All in all, we have succeeded in synthesizing new complexes, metal complexes and metal complex salts, which open up new possibilities in diagnostic and therapeutic medicine. In particular, the development of new imaging practices in medical diagnostics makes this development desirable.

In the following examples, the invention will be further elucidated.

The following describes the synthesis of starting material in the form of examples starting from a cyclization reaction: a) 1-Benzyl-4,7,10-tris- (p-tolylsulfonyl) -1,4,7,10-tetraaza-5 cvklododecan_

To a solution of 164.6 g of N, N ', N' '- tris- (p-tolylsulfonyl) diethylenetriamine-N, Ν'-disodium salt in 2.16 liters of dimethylformamide is dropped over 3 hours at 100 ° C under stirring 145 g of N, N-bis- [2,2 '- (p-tolylsulfonyloxy)] -ethane-benzylamine, which is dissolved in 900 ml of dimethylformamide. Then, with stirring at 80 ° C, 1 liter of water is added dropwise and further stirred for 18 hours at room temperature, then cooled to 0 ° C. The precipitate is aspirated, washed with slightly ice-cold ethanol and dried at 15 Torr and 60 ° C. 175 g of the title compound are obtained.

Alternative pathway 1

An analogous method of preparing tetraazacycloodecane derivatives is found in M. Hediger and T.A. Kaden, Helv. Chim. Acta 66. 861 (1983).

30.94 g of N, Ν ', N' '- tris (p-tolylsulfonyl) diethylenetriamine-20 N, N' 'disodium salt and 28.12 g of N-bis- (2-methanesulfonyloxyethyl) -triphenylamine The mixture is stirred together with 530 ml of dimethylformamide for 20 hours at 80-85 ° C, then cooled and stirred in a solution of 30 g of potassium carbonate in 5 liters of ice water. The precipitate is aspirated, the filter cake washed with 0.5 liters of water and dried at 20 ° C under vacuum at 150 Torr. To the purification, dissolve in 230 ml of chloroform and 5 ml of triethylamine, filter, evaporate under vacuum to 200 ml and add 250 ml of ethyl acetate to the solution at the boiling temperature. It is cooled overnight and the precipitated crystals are aspirated.

22.18 g of 1,4,7-tris- (p-tolylsulfonyl) -10-triphenylmethyl-1,4,7,10-tetraazacyclododecane are obtained, mp: 185-188 ° C (dec. 171574 B1 composing).

For the trityl protecting group decomposition, 31.4 g of the thus-obtained tritosyl-trityl derivative are stirred in a mixture of 100 ml glacial acetic acid, 75 ml water and 300 ml dioxane for 5 l hours at 80 ° C. Most are evaporated under vacuum at 60 ° C and diluted with 300 ml of ice water and 40 ml of 11 N sodium hydroxide solution (pH above 12) is added. This mixture is shaken with 300 ml of chloroform, the phases are separated, the aqueous phase is extracted twice, each time with 100 ml of chloroform, and the combined chloroform phases are dried over sodium sulfate and evaporated in vacuo.

The foamy residue is treated with 300 ml of diethyl ether, resulting in crystallization. Suction is dried, the crystals are dried under vacuum at 60 ° C and 150 Torr to give 21 g of 1,4,7-tris- (p-tolylsulfonyl) -1,4,7,10-tetraazacyclododecane, m.p .: 202-203 ° C.

The tritosyl compound thus obtained (21 g) is dissolved in 200 ml of dimethylformamide, to which solution is added successively 13.71 g anhydrous potassium carbonate, 4.95 g sodium iodide and 7.92 g benzyl bromide and stirred for 5 hours at 100 ° C. .

It is then cooled to 20 ° C, the mixture is stirred in 4 liters of ice water, suctioned and dissolved in 2 liters of dichloromethane. The solution is extracted with 100 ml of water, dried over sodium sulfate and evaporated under vacuum. The residue is dissolved in the boiling point temperature in 500 ml of acetonitrile, crystallized overnight, suctioned and dried, at 50 ° C and 150 Torr. 16.20 g of 1-benzyl-4,7,10-tris- (p-tolylsulfonyl) -1,4,7,7-tetraazacyclododecane are obtained, mp: 217-219 ° C.

b) N-benzyl-1,4,7,10-tetraazacyclododecane 150 g of 1-benzyl-4,7,10-tris- (p-tolylsulfonyl) -1,4,7,10-30 tetraazacyclododecane are heated together with 900 ml of HBr / glacial acetic acid (40%) and 125 g phenol for 16 hours at 50 ° C. After cooling to 20 ° C, dilute with 1 liter of ether, cool to -5 ° C and extract the precipitated crystals. To isolate the free base, the product is dissolved in 500 ml of 4 N sodium hydroxide solution, saturated with potassium carbonate and extracted several times with chloroform, dried over magnesium sulfate and evaporated in vacuo. 39 g of the title compound are obtained as a pale yellow 5 cool oil. A sample is characterized as trihydrochloride; mp 210 ° C (during decomposition).

Alternative pathway 2

To a solution of 11.2 g of 1,4,7,10-tetraazacyclododecane in 900 ml of tetrahydrofuran is added, at -20 ° C, 58 ml of triethylamine, 10 and within 3 hours a solution of 16.2 ml of benzoyl chloride is added dropwise. 280 ml of tetrahydrofuran, which easily raises the temperature above -10 ° C. The mixture is then stirred at 0-10 ° C for 16 hours, the precipitate is filtered off and the solution is evaporated in vacuo. The residue is chromatographed on 1 kg of silica gel and eluted with a dioxane-water-ammonia solution (8: 1: 1). After TLC, the uniform fractions are combined, evaporated, dissolved in dichloromethane and filtered to remove small haze, and the solution is evaporated. 19.60 g of 1,4,7-tribenzoyl-1,4,7,10-tetraazacyclododecane are obtained, mp: 120-20 125 ° C.

11.5 g of the tribenzoate thus obtained is stirred in 150 ml of dimethylformamide together with 8.3 g of anhydrous potassium carbonate, 3.0 g of sodium iodide and 7.2 ml of benzyl bromide for 18 hours at 100 ° C. Filter, evaporate the solution in vacuo, stir the residue twice, each time with 50 ml of hexane, and decant. The hexane phase is poured. For purification, dissolve in dichloromethane and chromatograph over 0.5 kg of silica gel with dichloromethane-methanol (37: 3). 10.2 g of 1,4,7-tribenzoyl-10-benzyl-1,4,7,10-tetraazacyclododecane are obtained, mp 105-1109 ° C.

To decompose the benzoyl group, this product (2.87 g) is dissolved in 290 ml of tetrahydrofuran, 11.2 g of potassium t-butylate is added and heated to reflux for 48 hours. File 24 DK 171574 B1 is evaporated, evaporated in vacuo, added 100 ml of water to the residue under ice-cooling and extracted three times, each time with 50 ml of dichloromethane. The combined dichloromethane phases were shaken with 10 ml of water, dried over sodium sulfate and evaporated under vacuum.

The initially oily residue crystallizes very slowly and is triturated with 20 ml of hexane. After extraction and drying, 1.15 g of N-benzyl-1,4,7,10-tetraazacyclododecane are obtained, m.p. : 75-78 ° C.

c) 1-Benzyl-4,7,10- (ethoxycarbonylmethyl) -1,4,7,10-tetraaza-cyclododecane

To a solution of 131.8 g of N-benzyl-1,4,7,10-tetraazacyclododecane in 1.5 liters of dichloromethane is added at 0 ° C in succession 200 g of triethylamine and 260 g of bromacetic acid ethyl ester over two hours. The mixture is stirred for another 16 hours at room temperature, shaken with a 5% sodium carbonate solution and brine, dried over magnesium sulfate and evaporated in vacuo. The residue is dissolved in 200 ml of chloroform and filtered through 2 kg of silica gel eluting with 1 liter of chloroform-methanol (95: 5). 210 g of the title compound are obtained as a cool oil.

d) Ν, Ν'Ν '- tris- (ethoxycarbonylmethyl) -1,4,7,10-tetraazacycloododecane 100 g of 1-benzyl-4,7,10-tris- (ethoxycarbonylmethyl) -1,4,7 10-tetraazacyclododecane is dissolved in 0.5 liters of acetic acid and 0.5 liters of ethyl acetate; 5 g of palladium-on-carbon (10%) is added and shaken for 5 hours under a hydrogen atmosphere. The catalyst is filtered off, evaporated under vacuum, the residue dissolved in 1 liter of chloroform, shaken with 100 ml of soda solution and 100 ml of brine, dried and evaporated under vacuum. The residue 30 is purified by ball-tube distillation at 10 ~ 3 Torr and 120 ° C. Ν, Ν ', N' '- tris- (ethoxycarbonylmethyl) -1,4,7,10-tetraazacyclododecane is obtained as a cool pale yellow oil.

DK 171574 B1

Yield: 65 g.

IR (film): 3400, 2935, 2878, 1738 cm -1.

Example 1

Gadolinium III complex of N- [2,3-dihydroxy-N-methyl-propyl-5-carbamoylmethyl] -1,4,7,10-tetraazacyclododecane-N ', Ν' ', Ν' '' triacetic acid 55 g N- [2,3-dihydroxy-N-methyl-propylcarbamoylmethyl] -Ν ', Ν' ', Ν' '-tris - (ethoxy carbonylmethyl) -1,4,7,10-tetraaza-cyclododecane is dissolved in 0 , 5 liters of ethanol, add 96 ml of 3 N 10 sodium hydroxide solution and stir for 3 hours at 20 ° C. Then, evaporate under vacuum, add 300 ml of water and adjust the pH to 6 with 2N hydrochloric acid. To this solution was added 31.94 g of gadolinium acetate and stirred for 18 hours at 50 ° C, then treated with an anion exchanger Amberlite IRA 410; Subsequently, the aqueous eluate is treated with a cation exchanger Amberlite IRC 50.

The eluate is evaporated under vacuum and dried. 47.14 g (73% of theory) of the title compound is obtained as a colorless powder.

Analysis: c20H34GdN5 ° 9 (645.77) C 37.20 H 5.31 Gd 24.35 N 10.85 (calculated) 37.52 5.19 24.09 10.87 (found)

The starting material for the preparation of the title compound of Example 2 is obtained as follows: a) N, Ν ', N' '- tris- (ethoxycarbonylmethyl) -1,4,7,10-tetraaza-cyclododecane-N' '- acetic acid

Dissolve 20 g of N, N ', N' - tris- (ethoxycarbonylmethyl) 1,4,7,10-tetraazacyclododecane in 300 ml of dichloromethane, add 10.40 g of triethylamine and then drop at 0 ° C a 26 DK 171574 B1 solution of 4.40 g of chloroacetic acid in 100 ml of dichloromethane and stir for 20 hours at room temperature. For processing, partition between dichloromethane and phosphate buffer, pH 6, dry over magnesium sulfate and evaporate under vacuum. 5 g of the desired compound are obtained as a cool oil.

b) N- [2,3-dihydroxy-N-methyl-propylcarbamoylmethyl] -Ν ', Ν' ', Ν' '' - tris - (ethoxycarbonylmethyl) -2,4,7,10-tetraazacy-clododecane

To a solution of 48.86 g (100 mmol) of N, Ν ', N' '- tris-10 (ethoxycarbonylmethyl) -1,4,7,10-tetraazacyclododecane-N' '-acetic acid in 500 ml of dichloromethane is added dropwise at 0 ° C 10.5 g of triethylamine and then a solution of 14 g of chloroformic acid isobutyl ester. Stir for 1 hour at 0 ° C, then drop a solution of 10.52 g of N-methylamino-2,3-propanediol in 100 ml of chloroform and stir for 2 hours at room temperature. The mixture is then shaken with a sodium bicarbonate solution and a saturated brine solution, dried over magnesium sulfate and evaporated in vacuo. For the purification, the chloroform solution of the residue is filtered through 500 g of silica gel to obtain 55 g of the title compound.

Example 2

Gadolinium-III Complex of N- [N-Ethylcarbamoylmethyl] - 1.4.7, 10-Tetraezacycloododecan-N'.Ν ''. Ν '' '- Acetic Acid

24 g of N- [N-ethylcarbamoylmethyl] -m ', N' ', N' '- tris-25 (ethoxycarbonylmethyl) -1,4,7,10-tetraazacyclododecane are dissolved in 250 ml of ethanol, 47 ml of 3N is added. of soda liquor and stir for 3 hours at 20 ° C, then evaporate under vacuum, add 150 ml of water and adjust the pH to 6 with 2N hydrochloric acid. 15.55 g of gadolinium acetate are added and stirred for 5 hours at 30 ° C. The solution is then purified as described in the preceding example by means of ion exchangers. 20.39 g of the title compound are obtained as a colorless powder.

Analysis: C 18 H 30 G <5 ° 7 (585.72) C 36.91 H 5.16 Gd 26.85 N 5.16 (calculated) 36.98 5.28 26.58 5.07 (found) 27 DK 171574 B1

Preparation of the starting material: 25 g (51.2 mmol) of N, Ν ', Ν' '- tris- (ethoxycarbonylmethyl) -1,4,7,10 -tetraazacyclododecane-N' '- acetic acid ( see Example 1) in 200 ml of dichloromethane, add 5.40 g of triethylamine at 0 ° C and then 7 g of chloroformic acid isobutyl ester. Stir for 1 hour at 0 ° C and then drop a solution of 2.31 g of ethylamine in 20 ml of dichloromethane, stir for 2 hours at room temperature, shake with a saturated sodium hydrogen carbonate solution and brine, dry over magnesium sulfate and evaporate to dryness. under vacuum. For the purification, the dichloromethane solution of the residue is filtered over silica gel over 200 g and 24 g of N- [N-ethylcarbamoylmethyl] -N, N '', N '' - tris- (ethoxycarbonylmethyl) -1,4,7,10-tetraazacy are obtained. -klododecan.

Example 3

Gadolinium III complex of N- (2,3-dihydro-1-propyl) -1,4,7,10-20 tetraazacycloododecan-N'.Ν.'.Ν.'.

4.30 g of N, Ν ', N' '- tris- (ethoxycarbonylmethyl) -1,4,7,1-tetraazacyclododecane is dissolved in 100 ml of dichloromethane, 4.20 g of triethylamine and 2.21 g of 3-chloro-methane are added. l, 2-propanediol.

Stir for 16 hours at room temperature, shake with water 25 and evaporate under vacuum. The residue is stirred for 6 hours with 100 ml of 1 N sodium hydroxide solution, the pH is adjusted to 6 with 2 N hydrochloric acid and stirred for 16 hours with 3.34 g of gadolinium acetate at 50 ° C. The solution thus obtained is subjected to an ion exchange purification to obtain 3.62 g of the title compound as a colorless powder.

Analysis: C17 H29 GdN4 O8 (574.69) C 35.53 H 5.09 Gd 27.36 N 9.75 (calculated) 35.68 5.19 27.03 9.68 (found) 28 DK 171574 B1

Example 4 Gadolinium-III Complex of N- (2-Aminoethyl) -1,4,7,10-tetra-azacycloododecan-N'ΝΝ ''Ν Acetic Acid

To a solution of 4.30 g of N, N ', N' '- tris- (ethoxycarbonylmethyl) -1,4,7,10-tetraazacyclododecane in 100 ml of dichloromethane is added 2.10 g of triethylamine and 1.22 g. g of N- (2-chloroethyl) acet-10 amide. Stir for 18 hours at room temperature, shake with water and evaporate under vacuum. The residue is stirred for 8 hours with 50 ml of 1N sodium hydroxide solution at 60 ° C, the pH is adjusted to 6 with 2N hydrochloric acid and stirred for 16 hours with 3.34 g of gadolinium acetate at 50 ° C. The solution thus obtained is subjected to an ion exchange purification and 3.21 g of the title compound are obtained as a colorless powder.

Analysis: C16 H28 G <+ N5 ° 6 (543.68) C 35.35 H 5.19 Gd 28.92 N 12.88 (calculated) 35.17 5.45 28.68 12.81 (found) Example 5

Bis-gadolinium III complex of 1,1 '- (1,3-propylene) bis (1,4,7,10-tetraazacycloododecan-4.7.10-acetic acid)

To a solution of 8.60 g of N, N ', N' '- tris- (ethoxycarbonylmethyl) -1,4,7,10-tetraazacyclododecane and 4.2 g of triethylamine in 2500 ml of dichloromethane are added 2, 02 g of 1,3-dibromopropane and stirred for 20 hours at room temperature, shaken with water and brine and evaporated under vacuum. The residue is filtered with dichloromethane over 150 g of silica gel and evaporated.

A cool oil is obtained, which is stirred with 60 ml of water and the pH is adjusted to 6 with 2N hydrochloric acid. After addition of 29.66 gadolinium acetate, the mixture is stirred for 16 hours at 50 ° C and the solution is purified by anion and cation exchangers.

6.56 g of the title compound is obtained as a colorless powder.

5 Analysis S ^ 31 ^ 50 ^ 2 ^ 8 ^ 12 (1041.28) C 35.76 H 4.84 Gd 30.20 N 10.76 (calculated) 35.71 4.58 29.94 10.88 (found)

Example 6

Bis-gadolinium III complex of succinyl-bis (1,4,7,10-tetra-azacycloododecan-4.7.10-acetic acid)

To a solution of 4.30 g of N, Ν ', N' '- tris- (ethoxycarbonylmethyl) -1,4,7,10-tetraazacyclododecane and 4.20 g of triethylamine in 100 ml of dichloromethane is added at 0 1.705 g succinic dichloride, dissolved in 20 ml of dichloromethane, is added dropwise, and then stirred for 1 hour at room temperature. It is equiped with a sodium bicarbonate solution and with brine and evaporated under vacuum. The residue is chromatographed on 100 g of silica gel with dichloromethane / ethyl acetate (0-30%).

A cool oil is obtained which is stirred for 4 hours with 40 ml of 1 N sodium hydroxide solution, then diluted with water (100 ml) and the pH is adjusted to 6 with 2 N hydrochloric acid. After the addition of 3.34 g of gadolinium acetate, the mixture is stirred at 50 ° C for 16 hours and the solution is purified by means of ion exchangers. 3.90 g of the title compound is obtained as a white powder.

Analysis: c32H4 8Gc2N8 ^ 14 (1083.28) C 35.48 H 4.47 Gd 29.03 N 10.34 (calculated) 35.31 4.18 28.81 10.57 (found)

The preparation and insulation of complexing agents are shown in the following examples. The isolated complexing agents are subsequently transferred, e.g. with various paramagnetic ions, to the 30 complexes.

Example 7 N- (2-hydroxyethyl) -1,4,7,10-tetraazacyclododecane-N ', N' ', N' '' triacetic acid A solution of 6.46 g of N, Ν ', N' is reacted. -tris- (ethoxy-carbonylmethyl) -1,4,7,10-tetraazacyclododecane in 100 ml of dichloromethane with 3.2 g of triethylamine and 1.45 g of 2-chloroethanol. Stir for 4 hours at room temperature, then shake with a sodium hydrogen carbonate solution and with 10 brine, dry over magnesium sulfate and evaporate in vacuo. The residue is stirred for 16 hours in 60 ml of 1 N sodium hydroxide solution. When 5 N hydrochloric acid is added, the pH is adjusted to 2.5. The resulting suspension is placed on an ion exchanger (DOWEX 50W-X4 in the H + form), eluted with water and then with 0.5 M NH 3 solution.

Evaporate under vacuum and isolate the title compound by adding ethanol and suction of the precipitate. 4.24 g of the title compound are obtained, the purity of which is checked by pH titration and elemental analysis.

Analysis: C 16 H 30 N 4 ° 7 (39 ° / 44) 20 C 49.22 H 7.75 N 14.35 (calculated) 49.48 7.83 14.09 (found)

Example 8 N- [N- (2-hydroxyethyl) -carbamoylmethyl] -1,4,7,10-tetraazacy-clododecane-N '. A solution of 12.50 g of N, N ', N' '- tris (ethoxycarbonylmethyl) - 1,4,7,10-tetraazacyclododecane-N' '- acetic acid in 250 ml of dichloromethane is reacted at 0 ° C with 5.22 g of triethylamine and then with 3.50 chloromyric acidobutyl ester. After 1 hour, 1.6 g of ethanolamine dissolved in 50 ml of dichloromethane is added dropwise, and stirred for 2 hours at room temperature, shaken with a soda solution and with brine, dried over magnesium sulfate and evaporated in vacuo. The residue is chromatographed with chloroform / acetone (10: 1) on 200 g of silica gel to give 11 g of N- (2-hydroxyethyl) -carbamoylmethyl-N ', N' ', N' '-trisethoxycar-5 bonylmethyl) -1,4,7,10 -1etraazacyclododecane.

This product is stirred in 100 ml of 1 N sodium liquor for 5 hours at room temperature, then the mixture is acidified with dilute hydrochloric acid to pH 2.5 and the suspension is purified on a cation exchanger (DOWEX 50W-X4), eluting with water and then with a 0.5 M NH 3 solution. The eluate is evaporated widely and, after the addition of ethanol, the title compound crystallizes, which is isolated by filtration. 7.2 g of the title compound are obtained, the purity of which is checked by titration and elemental analysis.

Analysis: C 18 H 33 N 5 ° 8 (447.49) C 48.31 H 7.43 N 15.65 (calculated) 48.20 7.48 15.49 (found)

Example 9

Gadolinium Complex of N- (morpholinocarbonylmethyl) -1,4,7,10-tetreazacycloododecan-N *, N * '. Ν' '* - Threaded Acid Acid 55.77 g N- (morpholinocarbonylmethyl) -Ν', Ν '', N Tris (ethoxycarbonylmethyl) -1,4,7,10-tetraazacyclododecane is hydrolyzed analogously to Example 1 with sodium hydroxide solution and complexed with gadolinium acetate. 45.19 g of the title compound 25 (72% of theory) is obtained as a white powder.

The starting material for the title compound is obtained, proceeding analogously to Example 16, but using morpholine instead of N-methylamino-2,3-propanediol.

Analysis: C20H32G <+ N5 ° 8 (627/75) C 38.27 H 5.14 Gd 25.04 N 11.16 (calculated) 38.02 5.09 24.83 11.35 (found) 32 DK 171574 B1

Example 10 Bis-gadolinium complex of 1,1 '- (2-hydroxy-1,3-propylene) -bis- (1,4,7,10-tetraazacycloododecane-4.7.10-triacetic acid)

To a solution of 43.05 g of N, N ', N' '- tris- (ethoxy-carbonylmethyl) -1,4,7,10-tetraazacyclododecane in 450 ml of dimethylformamide is added 4.63 g of epichlorohydrin. After 1 hour, 10 g of sodium iodide is added and heated for 24 hours to 80 ° C. Evaporate under vacuum, distribute the residue between water and chloroform, dry the chloroform phase over sodium sulfate and evaporate under vacuum. The residue is chromatographed on 1 kg of silica gel with dichloromethane - 10% acetone to give 27.5 g, 1 '- (2-15 hydroxy-1,3-propylene) bis- (1,4,7,10-tetraazacyclododecane). triacetic acid ethyl ester) as a cool oil.

9.17 g of the ester thus prepared are dissolved in 200 ml of ethanol and stirred with 30 ml of 3 N sodium hydroxide solution for 20 hours at room temperature, adjusted to pH 6 with hydrochloric acid and stirred for 20 hours at 60 ° C together with 6 68 g of gadolinium acetate. The solution is purified on anion and cation exchangers to give 10.05 g of the title compound as a white powder.

Analysis: ^ 31 ^ 5oG ^ 2 ^ 8G13 (1057.28) C 35.22 H 4.77 Gd 29.75 N 10.60 (Calc.) 25 35.03 4.89 29.49 10.41 (found) 33 DK 171574 B1

Example 11 N-methylglucamine salt of the manganese-II complex of N- [N- (2-hydroxyethyl) -carbamoylmethyl] -1,4,7,10-tetraazacyclododecan-N '.' '.' '' ' 8.95 g (20 mmol) of N- [N- (2-hydroxyethyl) -carbamoylmethyl] - 1.4.7.10- tetraazacyclododecane-N ', Ν' '- N' '' - acetic acid is suspended in 30 ml of water and heated together with 1.40 g (20 mmol) of manganese II oxide for 3 hours at 100 ° C. 3.90 g (20 mmol) of N-methylglucamine are then added, heated for an additional 12 hours to 100 ° C and the solution evaporated in vacuo to dryness. 13.8 g of the title compound are obtained as a pink-red powder, mp 140-143 ° C.

Analysis: c25H48MnN6 ° 13 (695.64) C 43.17 H 6.96 Mn 7.90 N 12.08 (calcd) 43.44 7.16 7.69 12.01 (found)

Example 12

Dysprosium III complex of N- (morpholinocarbonylmethyl) -1,4,7,10-tetraazacylclododecane-N, N ''. N '' - triacetic acid 20 g of N- (morpholinocarbonylmethyl) -Ν ', N' ', N' '- tris- (ethoxy-carbonylmethyl) -1,4,7,10-tetraazacyclododecane is hydrolyzed analogously to Example 1 with baking soda and complexed with dyspronium acetate. 16.3 g of the title compound are obtained as a white powder.

Analysis: c20H32DyN5 ° 8 (633.01) 25 C 37.95 H 5.10 Dy 25.67 N 11.06 (calculated) 37.71 4.92 25.81 11.32 (found) 34 DK 171574 B1

Example 13

Preparation of gadolinium N- (morpholinocarbonylmethyl) -1,4,7,10-tetraazacyclododecan-N ', N' ', N' '' - acetic acid-associated liposomes In accordance with that of Proc. Natl. Acad. Sci. U.S.A.

75. 4194 discloses a lipid mixture prepared from 75 mole percent egg phosphatidylcholine and 25 mole percent cholesterol as a dry substance. Of this, 500 mg is dissolved in 30 ml of diethyl ether and 3 ml of an aqueous 0.1 M solution of 10 N- (morpholinocarbonylmethyl) -1,4,7,10-tetraaza-N ', N' ', N' is added dropwise. The triadic acid gadolinium complex in an ultrasonic bath. The treatment is continued in an ultrasonic bath for another 10 minutes and evaporated under vacuum. The gel-like residue is suspended in a 0.125 M sodium chloride solution and centrifuged at 0 ° C repeated times at 20,000 g to separate the non-encapsulated gadolinium complex. The suspension is subjected to a vial freeze-drying. It is used as a colloidal dispersion in a 0.9% sodium chloride solution.

Example 14 20 Preparation of a solution of the yttrium-90 complex of the 1.4.7.10-tetraazacyclododecan-N, Ν ', N' ', N' '' tetraacetic acid conjugate with monoclonal antibodies

For a suspension of 4 mg of 1, 4,7,10-tetraazacyclododecan-N, Ν ', Ν' ', Ν' '' - tetraazacyclododecan-N, Ν ', N' ', N' '' - tetraacetic acid Into 1 ml of water are added 2 mg of N- (3-dimethylaminopropyl) -N'-ethylcarbodiimide hydrochloride and then 1 ml of a solution of 0.6 mg of monoclonal antibody (specific against melanoma antigen) dissolved in 0.05 molar sodium hydrogen carbonate buffer (pH 7.8). Stir for 2 hours at room temperature and dialyze against a 0.3 M sodium phosphate buffer. Then, 1 ml of yttrium-90 solution is added in acetate buffer, pH 6 (prepared according to Int. J. Appl. Radiat. Isot., Vol. 36 [1985], 35, and incubated for 24 hours at room temperature. . The solution is passed through a Sephadex G 25 column and the radioactive protein fraction is sterile filtered and filled into vials. Lyophilization results in a storage-stable solid composition.

Example 15

Indium-III complex of N- (morpholinocarbonylmethyl) -1,4,7,10-tetraazacycloododecan-N'.Ν '' .Ν '' '- acetic acid.

One proceeds analogously to Example 9 and complexes with radioactive 111 indium chloride. To investigate whether the metal ions are completely bound as chelate, the solution of the title compound is investigated by thin layer chromatography on silica gel plates in the methanol-water system (2: 1). Non-chelated metal ions are hereby viewed as a radioactive zone at the site of application.

If necessary, the chelation is completed by further addition of N- (morpholinocarbonylmethyl) -Ν ', Ν' ', Ν' '' - tris- (ethoxycarbonylmethyl) -1,4,7,10 -tetraazacyclododecane and subsequent ester cleavage.

Similarly, the gadolinium-153 complex of N- (morpholinocarbonylmethyl) -1,4,7,10-tetraazacyclododecane-N ', Ν' ', N' '' - acetic acid is obtained.

Example 16 a) 10- (2,3,4-Trihydroxybutyl) -1,4,7-tris-carboxymethyl-1,4,4,7,10-tetraazabicyclododecane 10.0 g (28.87 mmol) Tris-carboxymethyl-1,4,7,10-tetra-azacyclododecane (DO3A) is dissolved in 40 ml of water and the pH is adjusted to 13 with 5 N sodium hydroxide. A solution of 6.24 g (43.30 mmol) of 2- (2,2-dimethyl-1,3-dioxolan-4-yl) ethylene oxide (DE 3 150 917) in 10 ml of dioxane is added. and stir for 24 hours at room temperature. After dilution with 60 ml of water, extract three times with 50 ml of ether. The aqueous phase is adjusted to pH 2 with 10% hydrochloric acid and then evaporated. The residue is dissolved in water and transferred to a cation exchange column (IR 120). After rinsing with water, the ligand is eluted with a 0.5 N aqueous ammonia solution. The fractions are evaporated and the ammonium salt is collected in a little water and transferred to an anion exchange column (IRA 67). The water is first washed with water and then eluted with 0.5 N aqueous formic acid.

Evaporate in vacuo and dissolve the residue in a little hot methanol, then add acetone. Thereby, the title compound crystallizes.

11.31 g (87% of theoretical yield) of a white air-flowing powder is obtained (11.1% water analysis).

Analysis: (corrected for water) calculated: C 47.99 H 7.61 N 12.44 O 31.97 Found: C 47.93 H 7.67 N 12.40 b) Gadolinium complex of 10- (2, 3,4-Trihydroxybutyl-1,4,7-tris-carboxymethyl-1,4,7,10'-tetraazabicyclododecane 10.0 g (22.2 mmol) of the compound of Example 17 (a) is dissolved in 60 ml of demineralized of water, and the solution is added 4.02 g (11.1 mmol) of gadolinium oxide. It is heated to 90 ° C for 3 hours.

The cooled solution is stirred for 1 hour at room temperature with 2 ml of acidic ion exchanger (IR 120) and with 2 ml of basic ion exchanger (IRA 410). The ion exchanger is filtered off and the filtrate is briefly boiled with activated charcoal.

After filtration and freeze-drying, 12.76 g (95% of theoretical yield) of a white amorphous powder (12.3% water 37) is obtained according to analysis).

Analysis: (corrected for water) Calc'd: C 35.73 H 5.17 N 9.26 0 23.8 Gd 25.99 Found: C 35.68 H 5.24 N 9.21 Gd 25.93 Example 17 a) 10- (6-Hydroxy-2,2-dimethyl-1,3-dioxepan-5-yl) -1,4,7-tris-(p-toluenesulfonyl) -1,4,7,10-tetraazacycloododecane 50 g (78 (76 mmol) 4,7,10-tris- (p-toluenesulfonyl) -1,4,7,10-tetraazacyclododecane and 13.63 g (94.51 mmol) 4,4-dimethyl-3,5,8 -trioxabicyclo- (5.1.0) -octane is dissolved in 300 ml of dimethylformamide and heated to 170 ° C for 24 hours in an autoclave. Evaporate to dryness and chromatograph the residue over silica gel (solvent: methylene chloride / hexane / acetone: 10/5/1). The major fractions are evaporated and recrystallized from methyl tert-butyl ether / methanol. Yield: 52.76 g (86% of theoretical yield) of a cream powder.

Analysis: Calculated: C 55.51 H 6.47 N 7.19 S 12.35 Found: C 55.46 H 6.52 N 7.18 S 12.32 b) 10- (6-Hydroxy-2, 2-Dimethyl-1,3-dioxepan-5-yl) -1,4,7-tetrazacycloododecane 50 g (64.19 mmol) of the title compound of Example 18 (a) is suspended in 800 ml of liquid ammonia and 400 ml of tetrahydro -furan and the suspension is cooled to -35 ° C. Over 30 minutes, 8.9 g (1.28 moles) of lithium are added and stirred for 8 hours at -35 ° C. Excess lithium is destroyed by careful addition of methanol. Gently evaporate the ammonia gas and evaporate to dryness. The residue 38 DK 171574 B1 with 400 ml of hot toluene. The organic phases are dried over solid potassium hydroxide and then evaporated in vacuo. The residual oil is chromatographed over silica gel (solvent: methanol / water / concentrated ammonia solution = 10/1/1). 8.53 g (42% of theoretical yield) of pale yellow oil are obtained which solidifies on standing (8.1% water according to analysis).

Analysis (corrected for water): Calculated: C 56.93 H 10.19 N 17.71 Found: C 56.88 H 10.15 N 17.64 10 c) 10- (1-Hydroxymethyl-2,3-dihydroxypropyl ) -1,4,7-Tris-carb-oxy-methyl-1,4,7,10-tetraazacycloododecane (8 g) (25.28 mmol) of the title compound of Example 18 (b) is dissolved in 50 ml of water and the solution is added 14.05 g (101.12, 15 mmol) of bromoacetic acid. The pH of 9.5 is adjusted by means of 6 N potassium hydroxide.

It is heated to 50 ° C and the pH is maintained between 9.5 and 10 by the addition of 6 N potassium hydroxide. After 12 hours of stirring at 50 ° C, the solution is cooled in an ice bath, adjusted to pH 2 with concentrated hydrochloric acid and evaporated to dryness in vacuo.

The residue is dissolved in a little water and transferred to a cation exchange column (IR 120). After rinsing with water, the ligand is eluted with a 0.5 N aqueous ammonia solution.

The fractions are evaporated and the ammonium salt is collected by a little water and transferred to an anion exchange column (IRA 67). Wash with water first, then elute with 0.5 N aqueous formic acid.

After evaporation in vacuo, the residue is dissolved in slightly warm methanol and acetone is added. After cooling in an ice bath, the title compound crystallizes.

39 DK 171574 B1

Yield: 8.56 g (69% of theoretical yield) of a hygroscopic solid (9.1% water according to analysis).

Analysis (corrected for water): Calculated: C 51.42 H 7.81 N 11.42 Found: C 51.37 H 7.86 N 11.37 d) Gadolinium complex of 10- (1-hydroxymethyl-2 , 3-Dihydroxy-propyl) -1,4,7-tris-carboxymethyl-1,4,7,10-tetraazacyclo-dodecane (8 g) (16.31 mmol) of the title compound of Example 18 (c) is dissolved in 50 ml of deionized water and the solution is added 2.96 g (8.15 mmol) of gadolinium oxide. It is heated to 90 ° C for 3 hours. The cooled solution is stirred for 1 hour at room temperature, first with 2 ml of an acidic ion exchanger (IR 129) and then with 2 ml of a basic ion exchanger (IRA 410). The ion exchanger is filtered off and the filtrate is briefly boiled with activated charcoal. After filtration and freeze-drying, 9.99 g (95% of theoretical yield) of an amorphous powder (7.8% water according to analysis) is obtained.

Analysis (corrected for water): Calculated: C 39.12 H 5.47 N 8.69 Gd 24.39 Found: C 39.07 H 5.51 N 8.61 Gd 24.32

Example 18 a) 1- (2-Hydroxypropyl) -4,7,10-tris-carboxymethyl-1,4,7,10-25 tetraazabicyclododecane 20 g (54.74 mmol) 1,4,7-tris-carboxymethyl 1,4,7,10-tetraaza-bicyclododecane (DO3A) is dissolved in 40 ml of water. The pH of 13 is adjusted by means of 5 N sodium hydroxide. Then a solution of 6.7 g (115.47 mmol) of propylene oxide is added to 30 ml of dioxane and stirred for 24 hours at room temperature.

After dilution with 100 ml of water, extract three times with each 50 ml of ether. The aqueous phase is adjusted to pH 2 with 10% hydrochloric acid.

40 DK 171574 B1

Evaporate in vacuo and the residue is purified on ion exchange columns as described above (see Example 18 (c)). The major fractions are evaporated and recrystallized from methanol / acetone.

Yield: 16.58 g (71% of theory) of a glassy solid (7.8% water according to analysis).

Analysis (corrected for water): Calculated: C 50.48 H 7.97 N 13.85 Found: C 50.42 H 7.93 N 13.89 b) Gadolinium complexes of 1- (2-hydroxypropyl) - 4,7,10-Tris-carboxymethyl-1,4,7,10-tetraazacycloododecane 15 g (37.09 mmol) of the title compound of Example 19 (a) 15 is dissolved in 90 ml of deionized water and the solution is added 6.72 g (18). , 54 mmol) gadolinium oxide. It is heated to 90 ° C for 3 hours.

The cooled solution is stirred for 1 hour with 5 ml of an acid ion exchanger (IR 120) and 5 ml of a basic ion exchanger (IR 7410).

The ion exchanger is filtered off and the filtrate is briefly boiled with activated charcoal.

After filtration and freeze-drying, 19.89 g (96% of theoretical yield) of a white amorphous powder (7.3% water according to analysis) is obtained.

Analysis (corrected for water): Calculated: C 36.55 H 5.23 N 10.03 Gd 28.15 Found: C 36.48 H 5.28 N 10.11 Gd 28.09

Bis-gadolinium-III complexes of 1,1 '- (1,2-ethylene) -bis- (1-, 4,7,10-tetraazacycloododecane-4.7.10-acetic acid)

Example 19 41 DK 171574 B1

One proceeds by analogy with Example 5, but 1,2-di-5-bromethane is used instead of 1,3-dibromopropane.

The title compound is obtained as a colorless amorphous powder (7.4% water according to analysis).

Analysis (corrected for water): Calculated: C 35.08 H 4.71 N 10.91 Gd 30.62 10 Found: C 35.01 H 4.78 N 10.85 Gd 30.57

Example 20

Bis-gadolinium III complexes of 1,1 '- (1,5-pentylene) -bis- (1,4,7,10-tetraazacycloododecane-4.7.10-acetic acid)

You proceed analogously to Example 5, using 1.5-15 dibromopentane instead of 1,3-dibromopropane.

The title compound is obtained as a colorless amorphous powder (6.7% water according to analysis).

Analysis (corrected for water): Calculated: C 36.86 H 5.62 N 10.42 Gd 29.25 Found: C 36.81 H 5.59 N 10.44 Gd 29.19

Example 21

Bis-gadolinium III complexes of 1,1 '- (1,6-hexylene) -bis- (1,4,7,10-tetraazacycloododecane-4,7,10-triacetic acid)

An analogy to Example 5 is used, but 1,6-dibromohexane is used instead of 1,3-dibromopropane.

42 DK 171574 B1

The title compound is obtained as a colorless amorphous powder (8.1% water according to analysis).

Analysis (corrected for water): Calculated: C 37.69 H 5.21 N 10.34 Gd 29.03 Found: C 37.63 H 5.27 N 10.28 Gd 28.96

Example 22 a) 10- [1- (1,2-Dihydroxyethyl) -2,3,4-tris-hydroxybutyl] -1,4,7-tris-carboxymethyl-1,4,7,10-tetraazacycloododecane 27.27 g (103, 92 mmol) 3,4-anhydro-1,2: 5,6-bis-O- (1-methyl-10 ethylidene) mannitol and 10 g (28.86 mmol) of 1,4,7-tris-carboxylate. Boxymethyl-1,4,7,10-tetraazacyclododecane (DO3A) is dissolved in a mixture of 80 ml of n-butanol and 10 ml of water, after which the pH is adjusted to 13 by means of 6 N potassium hydroxide. The mixture is refluxed for 24 hours. Then, evaporate to dryness, collect the residue with 200 ml of water / 50 ml of methanol and extract twice with 100 ml of tert-butyl methyl ether twice. The aqueous solution is adjusted to pH 1 with 5 N hydrochloric acid and evaporated to dryness. The residue is boiled (extracted) with 200 ml of methanol / 80 ml of methylene chloride. Cool in an ice bath and filter the solution from the precipitated potassium chloride. The filtrate is evaporated to dryness and the residue is dissolved in 45 ml of water / 20 ml of methanol and transferred to a column of poly- (4-vinylpyridine). The product is eluted with a solution of ethanol and water (1: 3). After evaporation in vacuo, the residue is chromatographed on a reverse phase column (RP 18 / solvent = gradient of water and tetrahydrofuran). After evaporation of the main fraction, 5.45 g (37% of theoretical yield) of a highly hygroscopic glassy solid is obtained.

Analysis (calculated on anhydrous substance): calculated: C 47.05 H 7.50 N 10.97 Found: C 46.87 H 7.61 N 10.81 43 DK 171574 B1 b) Gadolinium complex of 10- [ 1- (1,2-dihydroxyethyl) -2,3,4-tris-hydroxybutyl] -1,4,7-tris-carboxymethyl-1,4,7,10-te-traazacycloododecane 5.40 g (10.58 of the title compound of Example 22 (a) 5 is dissolved in 50 ml of deionized water and the solution is added 1.92 g (5.29 mmol) of gadolinium oxide. It is heated to 90 ° C for 3 hours. The cooled solution is stirred for 1 hour with 3 ml of an acidic ion exchanger (AMB 252c) and 3 ml of a weak basic ion exchanger (IRA 67). The ion exchanger is filtered off and the freeze-dried filtrate is filtered off.

Yield: 6.82 g (97% of theory) of a colorless amorphous powder.

Analysis (calculated on anhydrous substance): Calculated: C 36.14 H 5.31 N 8.43 Gd 23.65 Found: C 36.03 H 5.45 N 8.31 Gd 23.50

Example 23 a) 10- (2,11-Dihydroxy-undecyl) -1,4,7-tris-carboxymethyl-1,4,4,7,10-tetraazacycloododecane 19.36 g (103.92 mmol) 1,2-epoxy-11 -hydroxyundecane and 10 g of 20 (28.86 mmol) 1,4,7-tris-carboxymethyl-1,4,7,10-tetraazacyclododecane (DO3A) are dissolved in a mixture of 50 ml of dioxane and 80 ml of water, and The pH is adjusted to 10 with 6N potassium hydroxide. Stir for 24 hours at 70 ° C. After evaporation to dryness, the residue was collected with 200 ml of water / 50 ml of methanol and extracted twice with 100 ml of tert-butyl methyl ether.

The aqueous solution is adjusted to pH 3 with 5 N hydrochloric acid and evaporated to dryness. The residue is boiled (extracted) with 200 ml of methanol / 80 ml of methylene chloride. One is cooled in an ice bath and the precipitated potassium chloride is filtered off.

The filtrate is evaporated in vacuo and the residue is dissolved in 45 ml of water / 20 ml of ethanol and the solution is then transferred to a column of poly- (4-vinylpyridine). The product is eluted with a solution of ethanol and water (1: 3). After evaporation in vacuo, the residue is chromatographed on a reverse phase column (RP 18 / solvent = gradient of water and tetrahydrofuran). After evaporation of the main fraction, 10.30 g (67% of theoretical yield) of a highly hygroscopic glassy solid is obtained.

Analysis (calculated on anhydrous substance): Calculated: C 58.37 H 9.08 N 10.52 Found: C 56.28 H 9.15 N 10.41 b) Gadolinium complex of 10- (2.11- Dihydroxy-undecyl) -1,4,7-tris-carboxymethyl-1,4,7,10-tetraazacycloododecane 10.0 g (18.77 mmol) of the title compound of Example 23 (a) is dissolved in 70 ml of deionized water and the solution is added. , 40 g (9.38 mmol) of gadolinium oxide. Heat to 90 ° C for 3 hours. The cooled solution is stirred for 1 hour with 3 ml of an acidic ion exchanger (AMB 252c) and 3 ml of a weak basic ion exchanger (IRA 67). The ion exchanger is filtered off and the filtrate is freeze-dried.

Yield: 12.64 g (98% of theory) of a colorless amorphous powder.

Analysis (calculated on anhydrous): Calculated: C 43.71 H 6.60 N 8.16 Gd 22.89 Found: C 43.79 H 8.80 N 8.05 Gd 22.73

Example 24 a) 10- (2-Methoxyethyl) -1,4,7-tris-carboxymethyl-1,4,7,10-tetraazacycloododecane 5 g (14.43 mmol) 1,4,7-tris-carboxymethyl 1,4,7,10-tetraaza-cyclododecane is dissolved together with 2.41 g (17.4 mmol) of 1-bromo-3-oxabutane in a mixture of 40 ml of dioxane and 60 ml of water. By adding 6N potassium hydroxide, the pH is adjusted to 10, then stirred for 24 hours at 70 ° C, evaporated in vacuo and the residue dissolved in a mixture of 100 ml of water and 20 ml of methanol. The solution is extracted twice with tert-butyl methyl ether and the aqueous solution is adjusted to pH 3 with 6N hydrochloric acid and evaporated, then the residue is boiled with a mixture of 100 ml of methanol and 40 ml of dichloromethane. The filtrate is evaporated in vacuo and dissolved in 20 ml of a mixture of water and ethanol (2: 1), after which the solution 10 is transferred to a column of 30 g of poly- (4-vinylpyridine). Elute with water / ethanol (2: 1), evaporate the solution and chromatograph the residue on an inverted phase column (RP 18 / solvent = gradient of water and tetrahydrofuran). After evaporation of the eluate, 3.44 g (59%) of a glassy solid is obtained.

Analysis (calculated on aqueous substance): calculated: C 50.48 H 7.97 N 13.85 found: C 50.08 H 8.34 N 13.80 b) Dysprosium complex of 10- (2-methoxyethyl) - 1,4,7-Tris-20-carboxymethyl-1,4,7,10-tetraazacycloododecane 3.10 g of the title compound of Example 24 (a) (water content: 4.9% corresponding to 7.29 mmol) are stirred with 1.361 g (3.65 mmol) dysprosium oxide in 35 ml of water at 90 ° C for 18 hours. The nearly clear solution is stirred for 1 hour with 1 ml of an acidic ion exchanger 25 (AMB 252c) and 1 ml of a weak basic ion exchanger (IRA 67). The ion exchanger is filtered off and the filtrate is freeze-dried.

Yield: 3.90 g (94.9%) of an amorphous powder.

Analysis (calculated on anhydrous substance): calculated: C 36.20 H 5.18 Dy 28.81 N 9.93 Found: C 36.77 H 5.36 Dy 28.66 N 10.05 46 DK 171574 B1 I Analogously to Examples 24 (a) and 24 (b), the praseodymium complex of 10- (2-tert.-butoxyethyl) -1,4,7-tris-carboxymethyl-1,4,7,10-tetraazacyclododecane is obtained. when in Example 24 (a), instead of 1-bromo-3-oxobutane, the compound uses 1-5 bromo-4,4-dimethyl-3-oxa-pentane and in Example 24 (b) instead of dysprosium oxide, the compound is used praseodymium oxide.

Example 25 a) 10- (2,3,4,5,6-pentahydroxy-n-hexyl) -1,4,7-tris-carboxy-methyl-1,4,7,10-tetraazacycloododecane 10 g (28.86 mmol) 1,4,7-tris-carboxymethyl-1,4,7,10-tetraazacyclododecane is dissolved in a mixture of 50 ml of water and 50 ml of dioxane. Add 6N potassium hydroxide to pH 11-11.5, then add 8.46 g (34.6 mmol) of 1,2-anhydro-3,4,5,6-di-0-isopropylidene -D-glucitol 15 (preparation: see Carbohydrate Research, Vol. 190, 313-316, 1989) and stir for 24 hours at 70-80 ° C. Evaporate in vacuo and dissolve the residue in a mixture of 200 ml of water and 40 ml of methanol, after which concentrated hydrochloric acid is added to a pH of 2 and heated to 60 ° C for 3 hours. After evaporation in vacuo, the residue is boiled with 100 ml of methanol. The filtrate is evaporated, the residue is dissolved in water and the solution is transferred to a column of 100 g of poly- (4-vinylpyridine). Elute with 300 ml of water, evaporate the eluate in vacuo and recrystallize the residue from methanol / acetic acid ethyl ester.

Yield: 10.31 g (70% of theory) of a white amorphous solid.

Analysis (calculated on anhydrous): Calculated: C 47.05 H 7.50 N 10.97 Found: C 46.90 H 7.31 N 11.08

Claims (9)

Yield: 6.18 g (95% of theoretical yield) of a white amorphous powder. Analysis: Calculated: C 36.14 H 5.31 Gd 23.65 N 8.43 Found: C 36.01 H 5.55 Gd 23.47 N 8.71 Patent Claim 1. Macrocyclic 1,4,7,10-tetraazacyclododecane derivatives, characterized in that they have the general formula I r2 R100C-s. > -v / (I) XN C) in Λ Λ of R 00C_ / N_ / \ _C00R 20 wherein each of the groups R1 is independently hydrogen or a metal ion equivalent, DK 171574 B1 R2 is a straight or branched, saturated or unsaturated car -hydrocarbon group having 1 to 16 carbon atoms, optionally substituted with 1-5 hydroxy or C 1 -C 4 alkoxy groups, or R 3 5 2 'R2 is -CH 2 -XN V where X represents carbonyl or a straight or branched alkylene group having 1 to 10 carbon atoms optionally substituted with 1-5 hydroxy or C1-C4 alkoxy groups, or X is a straight or branched alkylene group having 5 to 23 carbon atoms, interrupted by oxygen atoms and wherein R3 and R4 are independently hydrogen , a straight or branched alkyl group having 1 to 16 carbon atoms, optionally substituted with 1-5 hydroxy or C 1 -C 4 alkoxy groups, or R 3 and R 4 together with the nitrogen atom is a pyrrolidine, piperidine, morpholine or piperazine ring, or R 2 or R 3 is one via an alkylene those having 20 to 20 carbon atoms which optionally carry terminal carbonyl groups and are optionally interrupted by 1-4 oxygen atoms or which are substituted by 1-5 hydroxy, C1 -C4 alkoxy or carboxy-C1- C4-alkylene groups, second linked macrocycle of formula I 'R10OOC-v> -v / (") in Λ Λ 1 R 00C_ / N_ / \ _C00R 25 or R 2 represents B or CH 2 -COB which accumulate particularly extensively in the organ or organ moiety being examined, or in the tumor, and salts thereof with organic or inorganic bases or amino acids or with inorganic or organic acids. Compounds according to claim 1, characterized in that R 1 represents a hydrogen atom. Compounds according to claim 1, characterized in that at least two of the substituents R1 are metal ion equivalents of at least one metal having atomic numbers 21 to 29, 42, 44 or 57 to 70. Compounds according to claim 1, characterized in that at least two of the substituents R1 are metal ion equivalents of at least one radionuclide of an element having atomic numbers 27, 29, 31, 32, 38, 39, 43, 49, 64, 70 or 77. Compound according to claim 1, characterized in that it is selected from: N- [2,3-dihydroxy-N-methyl-propylcarbamoylmethyl] -1,4,7,10-tetraazacyclododecane-N ', N' ', N' '- triacetic acid, N- [N-ethylcarbamoylmethyl] -1 , 4,7,10-tetraazacyclododecan-N ', - N' ', N' '' - acetic acid, N- (2,3-dihydroxy-1-propyl) * 1,4,7,10-tetraazacyclododecan-N ' N-2-triacetic acid, N- (2-aminoethyl) -1,4,7,10-tetraazacyclododecan-N ', N' ', N' '- triacetic acid, N- (2) hydroxyethyl) -1,4,7,10-tetraazacyclododecane-N ', N' ', N' '' - triacetic acid, N- [N- (2-hydroxyethyl) -carbamoylmethyl] -1,4,7,10-tetraazacyclododecane-N ', N' ', N' '' - acetic acid, N- (morpholinocarbonylmethyl) -1,4,7 , 10-tetraazacyclododecan-N ', N' ', N' '' - acetic acid, and the complexes of these compounds with paramagnetic metals or radioisotopes. DK 171574 B1 Compound according to claim 1, characterized in that it is a gadolinium, manganese and / or dysprosium complex of 1,1 '- (1,3-propylene) bis (1,4,7,10-tetraazacyclododecane -4,7,10-5 thread acetic acid), succinyl bis (1,4,7,10-tetraazacyclododecane-4,7,10-thread acetic acid or 1,1'- (2-hydroxy-1,3-propylene) ) -bis - (1,4,7,10-tetraazacyclododecan-4,7,7-triacetic acid). Diagnostic agent, characterized in that it contains at least one metal complex of the general formula I according to claim 1, wherein at least two of the substituents R1 represent a metal ion equivalent, optionally together with usual gallic additives. Use of the compounds of claim 2 as complexes. Use of at least one metal complex of general formula I, wherein at least two of the substituents R1 represent a metal ion equivalent, for the preparation of agents for use in NMR, X-ray, ultrasound or radio diagnostics or for radiotherapy.