DE10133435A1 - New (ethylene)-(propylene)-triaminepentaacetic acid derivatives for preparing pharmaceutical agents - Google Patents

Abstract

(Ethylene)-(propylene)-triaminepentaacetic acid derivatives (I) or its salts, hydrates, esters, solvates, prodrugs, metabolites and/or stereoisomers are new. (Ethylene)-(propylene)-triaminepentaacetic acid derivative of formula X<1>OC-CH(R1)-N(CH-COX<1>)-CH(R1)-CH(R1)-N(CH(R1)(COX<1>))-C(R1)2-C(R1 )2-C(R)2-(C(R1)2)2-N(CH-COX<1>)-CH(R1)COX1 (I) or its salts, hydrates, esters, solvates, prodrugs, metabolites and/or stereoisomers are new. n = 0 or 1; R1 = functionalities for coupling with a biocompatible macromolecule or biomolecule; or non-coordinating substituents; R1+R1 = 5- or 6-membered ring; X<1> = OZ or N(R2); Z = H or metal ion equivalent; R = non-coordinating substituent. At least one of R1 is a functionality coupling with a biocompatible macromolecule or biocompatible and at least two of X1 are OZ. Independent claims are included for: (1) Conjugate of formula N(CH2-COOZ)2-A1-N(CH2-COOZ)-B-N(CH2-COOZ)2 (II); (2) Use of (I) for the production of (II) with a biomolecule; (3) Pharmaceutical agents containing at least one (I) or (II), optionally with the additives commonly used in galenicals; (4) Use of (I) or (II) for the production of agents for NMR diagnosis and radiodiagnosis or radiotherapy; (5) Reacting a compound of formula NH2-A-NH-B-NH2 (III) with Nu-CH2-COOZ' (IV) to obtain (I) or (II); if in (I), R1 is nitro then the compound is optionally reacted with a biomolecule to convert R1 into a group other than nitro; optionally removing the present protecting groups, followed by reacting with at least one metal oxide or metal salt and optionally completely or partially substituting acidic hydrogen atoms by cations of inorganic and/or organic bases, amino acids or amino acid amides; (6) A kit comprising (I) or (II); (7) Complex of (I) and a metal (M1); (8) A method (P1) of magnetic resonance imaging a patient involving administering a complex and a non-toxic carrier, adjuvant or other vehicles and generating a magnetic resonance image of at least a part of the patient; and (9) A method of radioimmunotherapy of a human involving applying the complex where the biocompatible macromolecule is a protein and the metal is an element having an atomic number of 26, 27, 29, 31, 32, 37-39, 43, 49, 62, 64, 70, 75, 77, 82 or 83. B = group of formula -C(R<2>)(R<3>)-C(R<4>)(R<5>)-C(R<6>)(R<7>)-(C(R<8>)(R<9>))n-; R<2> - R<9> = 1-6C alkyl (optionally mono- or disubstituted by OH and/or containing 1 or 2 O and aralkyl (the aryl radical is optionally substituted by alkyl or alkoxy)) or H; A1 = group of formula (V); Bio = radical of biomolecule; R<1>' = not defined. Two of R<2> - R<9> can be part of a 5 - 6 membered ring. Provided that 1 - 4 radicals of R<2> - R<9> are not hydrogen atoms. alpha and beta are bonded to any of the adjacent N atom.

Description

The invention relates to the articles characterized in the claims, d. H. (Ethylene) - (propylene) -triamine pentaacetic acid (EPTPA) derivatives, process for their Production and use thereof for the production of pharmaceutical agents for the NMR or radiodiagnosis or radiotherapy.

Nuclear Magnetic Resonance (NMR) is today a widely used method of medical diagnostics used for in vivo imaging, which can be used to visualize bodily vessels and tissues (including tumors) by measuring the magnetic properties of protons in the body. There are z. B. Contrast agents used by influencing certain NMR parameters of the body protons (eg., The relaxation times T ¹ and T ² ) cause a contrast enhancement in the resulting images or make these images readable. Above all, complexes of paramagnetic ions, such as. Gadolinium-containing complexes (eg Magnevist®) are used for shortening the relaxation times due to the effect of the paramagnetic ions.

The use of radiopharmaceuticals for diagnostic and therapeutic purposes is also known for a long time in the field of biological and medical research. In particular, radiopharmaceuticals are used to treat certain structures, such as For example, to represent the skeleton, organs or tissues. The diagnostic Application presupposes the use of such radioactive agents, which vary according to Accumulate application specifically in the structures in the patient being examined should. These locally enriching radioactive agents can then by means of suitable Detectors, such as scintillation cameras or other suitable Recording method, tracked, plotted or scanned. The distribution and relative intensity of the detected radioactive agent characterizes the location of a structure, where the radioactive agent is located, and can detect the presence of anomalies in Structures and functions, pathological changes, etc. represent.

Similarly, radiopharmaceuticals can be used as therapeutic agents to irradiate certain pathological tissues or areas. Such treatment requires the production of radioactive therapeutic agents that are specific Enrich structures, organs or tissues.

Both paramagnetic ions, such as. Bd: Gd ³⁺ , Mn ²⁺ , Cr ³⁺ , Fe ³⁺ and Cu ²⁺ as well as many metallic radionuclides can not be administered in free form as solutions since they are highly toxic. In order to make these ions suitable for in vivo use, they are usually complexed. For example, EP-A-0 071 564 describes, inter alia, the meglumine salt of the gadolinium (III) complex of diethylenetriamine pentaacetic acid (DTPA) as a contrast agent for NMR tomography. A preparation containing this complex was approved as the world's first NMR contrast agent under the name Magnevist®. This contrast agent is distributed extracellularly after intravenous administration and is renally excreted by glomerular secretion. Passage of intact cell membranes is practically not observed. Magnevist® is particularly well suited for the presentation of pathological areas (eg inflammation, tumors).

However, the known contrast agents and radiotherapeutics are not for everyone Use cases satisfactory. So many of these funds are distributed in the entire extracellular space of the body. In order to improve the efficiency of these funds in the To increase diagnostics and therapy, it tries to determine their specificity and selectivity For example, to target cells or desired areas and structures of the body increase. An improvement of these properties, for example, by coupling the Metal complexes to achieve biomolecules according to the "drug-targeting" principle. As Biomolecules offer antibodies, their fragments, hormones, growth factors and Substrates of receptors and enzymes (DE 195 36 780 A1).

In recent years, cravings are specific in diseased tissues accumulating diagnostics and therapeutics increased. When coupling the However complexing agents to selectively enriching substances is often observed that the complexing properties of the complexing agents deteriorate, so that it can lead to a weakening of the complex stability. This can cause it Problems come when a physiologically relevant proportion of the toxic metal ions released from the conjugate in vivo. It can also reduce the Specificity of the biomolecules come through the conjugate formation in the chelating agent.

In US 5,248,764 are DTPA derivatives and their chelates with radioactive Metal isotopes revealed. The target specificity of these derivatives is achieved by coupling the DTPA achieved via a carbonyl to a peptide. As a result, however, this carbonyl radical goes lost for the complexation of the metal ion, so that a lighter release of the toxic metal ions is to be feared.

DTPA derivatives with a reactive side group attached to the methylene carbon atom a carboxymethyl side chain is disclosed in EP-A-0 297 307. This has the advantage that none of the complex binding sites through the reactive side chain, by means of which the derivative can be coupled, for example, to a biomolecule, is blocked. On the other hand, the reactive side chain in this position a unwanted steric influence on the complex formation and thus the Exercise complexing constant.

For example, other DTPA derivatives that are reactive on an ethylene bridge, for example Have benzyl group and their second ethylene bridge is also substituted, are in U.S. Patent Nos. 4,831,175 and 5,124,471.

The chelating agent (ethylene) - (propylene) -triaminepentaacetic acid (EPTPA) has already been used in DE 29 18 842 A1 for complexing heavy metal ions such as iron and manganese in the bleaching of pulp usable in papermaking (wood pulp) where he described the removal of such ions from the aqueous, the pulp to facilitate containing system.

The use of the gadolinium (III) complex of EPTPA as an MRI contrast agent has been reported by Yun-Ming Wang, et al. in J. Chem. Soc., Dalton Trans., 1998, 4113-4118. In addition, this article surprisingly discloses to those skilled in the art that the Gadolinium (III) EPTPA complex comparable to the gadolinium (III) -DTPA complex Stability constant has. This was particularly surprising because of the ethylene bridges in DTPA each two adjacent nitrogen atoms in the complexing of the gadolinium ion with this form a sterically ideal 5-ring, whereas in EPTPA by introducing a propylene bridge a sterically less ideal 6-ring with the central gadolinium ion must be formed.

It is further desirable to have means for diagnosis and therapy available which have the highest possible target specificity and the highest possible vivo stability for the most toxic metal ions possess.

An object of the invention was, therefore, new means for NMR and Radiodiagnostik as well as to provide the radiotherapy, the disadvantages mentioned not and in particular a high in vivo stability, good compatibility and above all have organ-specific properties. On the one hand, the retention in the zu be sufficient to allow for a low dose to obtain unambiguous diagnosis necessary number of pictures, on the other hand but should then as fast and as far as possible complete elimination of Be assured of metals from the body. Furthermore, the NMR contrast agents should have a high Show proton relaxivity and thus an increase in signal intensity Allow dose reduction.

It has now been found that this problem is solved by conjugates of biomolecules with (ethylene) - (propylene / butylene) -Triaminpentaessigsäure derivatives, whose ethylene bridge is substituted with a reactive benzyl group and whose propylene or butylene bridge has additional substituents. The invention thus relates to compounds of general formula I.


wherein
Z is a hydrogen atom or a metal ion equivalent,
A is a radical of the formula


stands, in which the with


labeled positions α and β are attached to any of the adjacent nitrogen atoms, R ^{1 is} a nitro group or a group capable of reacting with a biomolecule, and
B is a radical of the formula


in which n is 0 or 1 and R ² , R ³ , R ⁴ , R ⁵ , R ⁶ , R ⁷ , R ⁸ and R ^{9 are} independently selected from a hydrogen atom, a straight-chain or branched, saturated or unsaturated C _{1 -6-} alkyl group which may be optionally substituted with 1 or 2 hydroxy groups and / or may contain 1 or 2 oxygen atoms, and an aralkyl group, the aryl group may optionally be substituted by an alkyl or alkoxy group, wherein two of the radicals R ² , R ³ , R ⁴ , R ⁵ , R ⁶ , R ⁷ , R ⁸ and R ^{9 may be} part of a 5- or 6-membered ring, with the proviso that at least one and at most four of the radicals R ² , R ³ , R ⁴ , R ⁵ , R ⁶ , R ⁷ , R ⁸ and R ^{9 are} not hydrogen atoms, and their salts, preferably with inorganic or organic bases.

The compounds of the invention are due to the reactive benzyl to the Formation of conjugates with biomolecules, so that they easily organ-specific Lend properties and these can be easily varied. Despite the substituted Propylene and Butylenbrücke have metal complexes with the inventive Compounds high in vivo stability. In addition, the own Compounds according to the invention and their conjugates with biomolecules a good Compatibility and good water solubility, so that they can be used as pharmaceutical agents particularly suitable for NMR and radiodiagnosis and radiotherapy. The Relaxivity of the complexes of the invention is surprisingly high, so that the Complexes containing a paramagnetic ion are especially good for NMR spectroscopy. Diagnostics are suitable.

In the compounds of the formula I according to the invention A is a radical of the formula

This rest can be connected to the


labeled positions α and β be bound to any of the adjacent nitrogen atoms, ie the benzyl substituent of this radical may be adjacent to one of the two nitrogen atoms to which the radical A is attached. Preferably, however, the radical A is bonded via the position α to the (ZOOC-CH ₂ ) ₂ -N radical, so that the benzyl substituent is adjacent to this radical.

The phenylene group of the benzyl substituent of residue A is substituted with a nitro group or a group capable of reacting with a biomolecule. This substituent R ¹ is preferably bonded in the metha or para position, in particular in the para position, to the phenylene groups. The compounds of the formula I in which R ^{1 is} a nitro group are particularly suitable as intermediates for the preparation of those compounds of the formula I in which R ^{1 is} a group which is able to react with a biomolecule.

Suitable groups that can react with a biomolecule are, for example, amino (-NH ₂ ), isocyanate (-NCO), isothiocyanate (-NCS), hydrazine (-NHNH ₂ ), semicarbazide (-NHCONHNH ₂ ), thiosemicarbazide (-NHCSNHNH ₂ ), chloroacetamide (-NHCOCH ₂ Cl), bromoacetamide (-NHCOCH ₂ Br), iodoacetamide (-NHCOCH ₂ I), acylamino such as acetylamino (-NHCOCH ₃ ), maleimide, maleimidacylamino such as 3- (2.5 Dioxo-2,5-dihydro-pyrrol-1-yl) -propionylamino, activated esters, such as


mixed anhydrides, azide, hydroxide, sulfonyl chloride and carbodiimide.

B in formula I is a radical of the formula

Here n can be either 0 or 1, with compounds in which n = 0 being preferred. The substituents R ² , R ³ , R ⁴ , R ⁵ , R ⁵ , R ⁷ , R ⁸ and R ⁹ are independently selected from a hydrogen atom, a straight or branched, saturated or unsaturated C _1-6 alkyl group, optionally may be substituted by 1 or 2 hydroxy groups and / or may contain 1 or 2 oxygen atoms, and an aralkyl group whose aryl radical may optionally be substituted by an alkyl or alkoxy group. In this case, at least one and at most four of these radicals need not be hydrogen atoms, so that the propylene or butylene bridge B carries in the compounds of the formula I according to the invention at least one and a maximum of four substituents.

The substituent (s) of the bridge B may be a C _1-6 alkyl group which may be optionally substituted with 1 or 2 hydroxy groups and / or may contain 1 or 2 oxygen atoms. Preferably, the C _1-6 alkyl group is a methyl, ethyl, n-propyl, iso-propyl, n-butyl, iso-butyl or tert-butyl group. Simultaneously or alternatively, one or more substituents of bridge B may be an aralkyl group, with arylC _1-6 alkyl groups and in particular benzyl being preferred. The aryl radical of these aralkyl groups may be substituted with an alkyl or alkoxy group, preferably in the para position. Preferably, this alkyl group is a C _1-6 alkyl group, such as in particular methyl or ethyl, and in this alkoxy group to a C _1-6 alkoxy, in particular methoxy or ethoxy.

Preferably, the bridge B is substituted with 1 or 2 methyl, ethyl or benzyl groups.

Also preferred are such compounds of the invention wherein R ² , R ³ , R ⁴ , R ⁵ , R ⁵ , R ⁷ , R ⁸ and R ⁹ are selected such that B is symmetrical.

Additionally or alternatively, two of R ² , R ³ , R ⁴ , R ⁵ , R ⁶ , R ⁷ , R ⁸ and R ^{9 may be} part of a 5- or 6-membered ring. In the number of ring members of such a ring, the carbon atoms to which the ring-forming radicals R ² , R ³ , R ⁴ , R ⁵ , R ⁶ , R ⁷ , R ⁸ or R ^{9 are} attached, as well as optionally intervening carbon atoms of the Propylene or butylene bridge B with. The ring may be saturated or unsaturated, with 5- and 6-membered saturated rings being preferred.

Preferred propylene or butylene bridges B are: -CH ₂ -CH ₂ -CH (CH ₂ -CH ₃ ) -, -CH (CH ₂ - CH ₃ ) -CH ₂ -CH ₂ -, -CH ₂ -C (CH ₃ ) ₂ -CH ₂ -, -CH ₂ -CH (CH ₃ ) -CH ₂ -, -CH ₂ -CH (CH ₂ -phenyl) -CH ₂ -, -CH ₂ -CH (CH ₃ ) -CH ( CH ₃ ) -CH ₂ -,

The compounds of the invention contain at least one chiral center. Also if in the description and the claims not between the different Enantiomers, include the compounds mentioned, if not otherwise stated, always both enantiomers and in the presence of multiple stereocenters also all possible diastereomers and mixtures thereof.

The compounds of the formula I according to the invention are suitable for the preparation of conjugates with biomolecules. These conjugates have the general formula II


wherein Z and B are as defined above and A 'is a radical of the formula


stands, in which the with

characterized positions α and β are bound to any of the adjacent nitrogen atoms and Bio represents the remainder of a biomolecule which is bound via the radical R ^{1 'of} a reactive group to the phenylene ring, and their salts, preferably with inorganic or organic bases. The radical R ^{1 '} is preferably a radical R ¹ as defined above after its reaction with a biomolecule.

By "biomolecule" is meant herein any molecule that either naturally For example, occurs in the body or was prepared synthetically with analog structure. In addition, these are understood to mean those molecules which are biologically, For example, occurring in the body molecule or occurring there structure in Interaction, so that, for example, the conjugates at certain, Enrich desired parts of the body. Under "body" in the present case everyone understood herbal or animal body, with animal and in particular human bodies are preferred.

To form conjugates with the compounds of the invention are the following Biomolecules particularly suitable.

Biopolymers, proteins, such as proteins that have a biological function, HSA, BSA, etc., Proteins and peptides that accumulate in certain places in the organism (eg Receptors, cell membranes, channels, etc.), peptides cleavable by proteases, peptides with synthetic breaking points (eg labile esters, amides, etc.), peptides by Metalloprotheasen be cleaved, peptides with photocleavable linkers, peptides with oxidative (oxydases) cleavable groups, peptides with natural and unnatural amino acids, glycoproteins (glycopeptides), signaling proteins and antiviral Proteins, synthetically modified biopolymers, such as linker-derivatized biopolymers, modified metalloproteases and derivatized oxidase, etc., carbohydrates (mono- to Polysaccharides), such as derivatized sugars, fungal sugars, cyclodextrins in the organism and its derivatives, amino sugars, chitosan, polysulfates and acetylneuraminic acid Derivatives, antibodies, such as monoclonal antibodies, antibody fragments, polyclonal Antibodies, Minibodies, Single Chains (even those with Linkem to multiple Fragments are linked), red blood cells and other blood components, Cancermarker (eg CAA) and cell adhesion substances (eg Lewis X and anti-Lewis X- Derivatives), DNA and RNA fragments such as derivatized DNAs and RNAs (e.g. found by the SELEX method), synthetic RNA and DNA (also with unnatural bases), PNAs (Hoechst) and antisense, β-amino acids (Seebach), Vector amines for introduction into the cell, biogenic amines, pharmaceuticals, oncological Preparations, synthetic polymers directed to a biological target (eg receptor) are, steroids (natural and modified), prostaglandins, taxol and its derivatives, Endothelins, alkaloids, folic acid and its derivatives, bioactive lipids, fats, fatty acid esters, synthetically modified mono-, di- and triglycerides, liposomes attached to the surface are derivatized, micelles from natural fatty acids or from perfluoroalkyl compounds, Porphyrins, texaphrins, extended porphyrins, cytochromes, inhibitors, neuraminases, Neuropeptides, immunomodulators such as FK 506, CAPE and gliotoxin, endoglycosidases, Substrates activated by enzymes such as calmodoline kinase, casein kinase II, Glutathione-S-transferase, heparinase, matrix metalloprotheases, β-insulin receptor Kinase, UDP-galactose 4-epimerase, fucosidases, G-proteins, galactosidases, Glycosidases, glycosyltransferases and xylosidase, antibiotics, vitamin and vitamin Analogs, Hormones, DNA Intercalators, Nucleosides, Nucleotides, Lectins, Vitamin B12, Lewis X and relatives, psoralens, dienriene antibiotics, carbacyclins, VEGF (vascular endothelial growth factor), somatostatin and its derivatives, biotin derivatives, antihormones, Dendrimers and cascade polymers and their derivatives, tumor specific proteins and Synthetics, polymers that accumulate in acidic or basic areas of the body (pH-controlled distribution), myoglobins, apomyoglobins etc., neurotransmitter peptides, Tumor necrosis factors, peptides that accumulate in inflamed tissue, blood pool Reagents, anions and cation transporter proteins, polyesters (eg lactic acid), Polyamides and polyphosphates.

Most of the aforementioned biomolecules are commercially available, for example, from Merck, Aldrich, Sigma, Calibochem and Bachem available.

In addition, as biomolecules all in WO 96/23526 and WO 01/08712 disclosed "plasma protein binding groups" or "target binding groups" used become. The content of these two publications is therefore incorporated by reference the present description was included.

Furthermore, the compounds of the invention are suitable for conjugation to all of them Molecules which are reacted with fluorescent dyes in the prior art to for example, their localization by epifluorescence microscopy within the cell too determine. Also, the compounds can be used with any drugs be conjugated to transport within then after administration of the drug of the organism, for example, by the NMR technique. It is also possible that the conjugates of the compounds of the invention and the biomolecules contain additional additional molecules that have been conjugated to the biomolecules. With The term "biomolecule" in the sense of the invention thus includes all molecules which are in biological systems and all molecules that are biocompatible.

The compounds of general formula I and their conjugates with biomolecules, for example, by reacting a compound of formula III

H ₂ NA-NH-B-NH ₂ III

wherein A and B are as defined above, with a compound of formula IV

Nu-CH ₂ -COOZ'IV,

where Nu is a nucleofuge and Z 'is a hydrogen atom, a metal ion equivalent, preferably an alkali or alkaline earth metal such as in particular sodium or potassium, or a protective group for carbonyl. The compound thus obtained can then be reacted with a biomolecule, the remainder of R ¹ , when it is nitro, first having to be converted into a group capable of undergoing reaction with a biomolecule. Subsequently, after removal of any protective groups still present and in a manner known per se, if desired, reaction can be carried out with at least one metal oxide or metal salt in order to obtain the desired metal complexes. If desired, any remaining acidic hydrogen atoms in the complexes thus obtained can then be wholly or partly substituted by cations of inorganic and / or organic bases, amino acids or amino acid amides.

The nucleofug used are advantageously the radicals:
Cl, F, -OTs and -OMs.

The reaction is carried out in a mixture of water and organic solvents such as:
Isopropanol, ethanol, methanol, butanol, dioxane, tetrahydrofuran, dimethylformamide, dimethylacetamide, formamide or dichloromethane performed. Ternary mixtures of water, isopropanol and dichloromethane are preferred.

The reaction is carried out in a temperature range between -10 ° C and 100 ° C, preferably between 0 ° C and 30 ° C performed.

The neutralization of any remaining free carboxy groups is carried out with the help inorganic bases (eg, hydroxides, carbonates or bicarbonates) of e.g. Sodium, Potassium, lithium, magnesium or calcium and / or organic bases, among others primary, secondary and tertiary amines, such as. Ethanolamine, morpholine, glucamine, N- Methyl and N, N-dimethylglucamine, as well as basic amino acids, such as. Lysine, arginine and ornithine or amide original neutral or acidic amino acids.

For the preparation of the neutral complex compounds can be, for example, in acid Complex salts in aqueous solution or suspension as much of the desired base Add that the neutral point is reached. The resulting solution can subsequently in Vacuum be evaporated to dryness. It is often beneficial to educate Neutral salts by adding water-miscible solvents such. B. lower Alcohols (methanol, ethanol, isopropanol and others), lower ketones (acetone and others), polar ethers (tetrahydrofuran, dioxane, 1,2-dimethoxyethane and others) precipitate and so easy to isolate and easy to clean crystals. As It has proven particularly advantageous already during the Add complex formation of the reaction mixture and thereby a process step save.

The preparation of the compounds of the formula I according to the invention is explained below using the example of a preferred compound in which R ^{1 is} -NO ₂ and B is -CH ₂ -CH ₂ -CH (-CH ₂ -CH ₃ ) - and Z is hydrogen. The representation of the connection


can be from the t butyl esters 1


by acid hydrolysis with trifluoroacetic acid. The compound of formula 1 may be obtained by alkylation of the amine of formula 2


be obtained with bromoacetic acid t-butyl ester. The ester is available from Merck, Fluka or Aldrich.

The amine 2 can be obtained by hydrolysis with trifluoroacetic acid from the protected compound 3


to be obtained.

Compound 3 is accessible by alkylation of mesylate 4 with 1,3-diaminopentane 5 and chromatographic separation of the amine mixture:

The amine 5 is commercially available (Aldrich, Fluka).

The mesylate 4 can be obtained by reacting the alcohol 6 with methanesulfonyl chloride in the presence of triethylamine:

The alcohol 6 can be obtained by reduction of the ester 7 with sodium borohydride in tetrahydrofuran / methanol (8: 1):

The 4-Nitrophenylalaninmethylester 7 can be prepared from the corresponding acid 8 by esterification with methyl iodide in the presence of sodium bicarbonate in dimethylformamide:

The acid 8 is commercially available (Aldrich, Fluka).

Alternatively, it is also possible to proceed in such a way that the building block 3 is formed by amide formation the phenylalanine 8 and the 1-ethyl-1,3-propanediamine 5 and subsequent reduction the amide bond is obtained. The chromatographic separation can be avoided when the amine component is used as a 3-N-BOC derivative.

The α, ω-diamines required for the synthesis are obtainable, for example, via the synthetic routes shown schematically below: C-3 diamines, substituents on C-2

R: alkyl etc.
R ': hydrogen (then step 3 of the first stage is omitted) or R
X: halogen C-3 diamines, substituent at C-1

C4 diamines, substituent at C-4

The conversion of the compounds of general formula I, in which R ¹ = NO ₂ , in compounds of general formula I, in which R ^{1 is} not equal to nitro, is carried out by known methods. Thus, the hydrogenation of the nitro group to the amino group succeeds, for example, with 10% palladium on carbon (compare EP 175 617, EP 173 629 and US Pat. No. 5,087,696).

The amino group may be prepared by reacting with nitrophenyl or hydroxysuccinimide esters in the corresponding amide are transferred. But you can also by reacting with Thiophosgen be converted into the isothiocyanate directly with amino groups for Thiourea couples. The isothiocyanate can also react with hydrazine to thiosemicarbazide react, then specifically with the anoxidized sugar molecules of an antibody converted to thiosemicarbazide.

In an analogous manner, the amine with phosgene to the isocyanate and this with hydrazine for Semicarbazone implemented. The anilino group is also acylatable. Will the implementation with carried out the Akitvester of 4-Maleimidobuttersäure (Fluka), one obtains a specific to -SH group binding reagent. An -SH groups also bind the Haloacetamides obtainable by reacting the anilines with halogenated activators.

The amino group itself is also anoxidierter as a binding site for the carbonyl groups Sugar usable if the partner molecule the conditions of reductive amination tolerated.

The preparation of complexes for the production of NMR diagnostic agents can be done in the way take place, as described in the patents EP 71564, EP 130934 and DE-OS 34 01 052 has been disclosed. For this purpose, the metal oxide or a metal salt (for example, a Chloride, nitrate, acetate, carbonate or sulfate) of the desired element in water and / or a lower alcohol (such as methanol, ethanol or isopropanol) or dissolved suspended and with the solution or suspension of the equivalent amount of reacted complexing agent according to the invention.

Should the complexing agents for the production of Radiodiagnostika or -therapeutika Use can, the production of complexes from the complexing agents according to the Radiotracers for Medical Applications, Vol. I, CRC Press, Boca Raton, Florida done methods described.

It may be desirable to prepare the complex shortly before use, especially if it is to be used as a radiopharmaceutical. Therefore, the Invention also provides a kit for the preparation of radiopharmaceuticals comprising a compound of formula I or a conjugate of formula II, wherein Z is hydrogen, and a compound a desired metal.

The invention furthermore relates to pharmaceutical agents which contain at least one physiologically acceptable compound of general formula I or at least one Physiologically acceptable conjugate of general formula II, optionally with the usual additives in galenics.

The preparation of the pharmaceutical agents according to the invention takes place in itself known manner, by the complex compounds of the invention - optionally with the addition of the additives customary in galenics - in an aqueous medium suspended or dissolved and then the suspension or solution optionally sterilized. Suitable additives are, for example, physiologically acceptable buffers (such as z. Tromethamine), additions of complexing agents or weak complexes (such as eg. Diethylenetriaminepentaacetic acid or the metal complexes of the invention corresponding Ca complexes) or - if necessary - electrolytes such. B. Sodium chloride or - if necessary - antioxidants such. As ascorbic acid.

Are for enteral administration or other purposes suspensions or solutions of agents according to the invention in water or physiological saline solution are desired with one or more excipients customary in galenicals [e. Methylcellulose, Lactose, mannitol] and / or surfactant (s) [e.g. Lecithins, Tween®, Myrj®] and / or Flavoring agent (s) for flavor correction [e.g. B. essential oils] mixed.

In principle, it is also possible to use the pharmaceutical compositions according to the invention without Isolation of the complex salts produce. In any case, special care must be taken be used to carry out the chelation so that the salts of the invention and saline solutions are virtually free of uncomplexed toxic metal ions.

This can be done, for example, by means of color indicators such as xylenol orange Control titrations are ensured during the manufacturing process. The invention therefore also relates to processes for the preparation of the complex compounds and their salts. As last certainty remains a purification of the isolated complex salt.

• 1. für die NMR-Diagnostik in Form ihrer Komplexe mit den pharmazeutischen Ionen der Elemente mit den Ordnungszahlen 21-29, 42, 44 und 58-70. Geeignete Ionen sind beispielsweise das Chrom(III)-, Eisen(II)-, Cobalt(II), Nickel(II)-, Kupfer(II)-, Praseodym(III)-, Neodym(III)-, Samarium(III)- und Ytterbium(III)-ion. Wegen ihres starken magnetischen Moments sind für die NMR-Diagnostik besonders bevorzugt das Gadolinium(III)-, Terbium(III)-, Dysprosium(III)-, Holmium(III)-, Erbium(III)-, Mangan(II)- und Eisen(III)-ion.
• 2. für die Radiodiagnostik und Radiotherapie in Form ihrer Komplexe mit den Radioisotopen der Elemente mit den Ordnungszahlen 26, 27, 29, 31, 32, 37-39, 43, 46, 47, 49, 61, 62, 64, 67, 70, 71, 75, 77, 82 und 83.

The pharmaceutical compositions according to the invention preferably contain 1 mmol-1.3 mol / l of the complex salt and are usually dosed in amounts of 0.0001-5 mmol / kg. They are intended for enteral and parenteral administration. The complex compounds according to the invention are used

• 1. for NMR diagnosis in the form of their complexes with the pharmaceutical ions of the elements with atomic numbers 21-29, 42, 44 and 58-70. Examples of suitable ions are the chromium (III), iron (II), cobalt (II), nickel (II), copper (II), praseodymium (III), neodymium (III), samarium (III) and ytterbium (III) ion. Because of their strong magnetic moment, the gadolinium (III), terbium (III), dysprosium (III), holmium (III), erbium (III), manganese (II) and iron (III) ion.
• 2. for radiodiagnostics and radiotherapy in the form of their complexes with the radioisotopes of elements with atomic numbers 26, 27, 29, 31, 32, 37-39, 43, 46, 47, 49, 61, 62, 64, 67, 70 , 71, 75, 77, 82 and 83.

The compositions of the invention meet the diverse requirements for suitability as Contrast agent for magnetic resonance imaging. So they are great for, after oral or parenteral administration by increasing the signal intensity with the help of the Magnetic resonance imaging obtained image in its expressiveness to improve. Further show They have the high potency that is necessary to keep the body in as small amounts as possible to contaminate foreign materials, and the good compatibility that is necessary to the to maintain non-invasive character of the investigations.

The good water solubility and low osmolality of the agents according to the invention allowed to produce highly concentrated solutions so that the volume load of the circuit in acceptable limits and dilution by the body fluid that is, NMR diagnostic agents must be 100 to 1000 times more water-soluble be as for NMR spectroscopy. Furthermore, the compositions of the invention do not have only a high stability in vitro, but also a surprisingly high stability in vivo, such that release or replacement of the non-covalently bound in the complexes - in itself poisonous - ions within the time in which the new contrast agents completely be excreted again, only very slowly.

In general, the agents according to the invention are used for the application as NMR Diagnostics in amounts of 0.0001-5 mmol / kg, preferably 0.005-0.5 mmol / kg, dosed. Details of the application are z. In H.-J. Weinmann et al., Am. J. of Roentgenology 142, 619 (1984).

Low doses (below 1 mg / kg body weight) of organ-specific NMR Diagnostics can be used, for example, to detect tumors and myocardial infarction. Particularly low dosages of the complexes of the invention are for the Application in radiotherapy and radiodiagnosis suitable.

Furthermore, the complex compounds according to the invention can be advantageously used as Susceptibility reagents and as shift reagents for in vivo NMR spectroscopy be used.

The agents of the invention are due to their favorable radioactive properties and the good stability of complex compounds contained in them as well Radiodiagnostika and radiotherapeutics suitable. Details of their application and dosage be z. In Radiotracers for Medical Applications, CRC-Press, Boca Raton, Florida. 1983, as well as in Eur. J. Nucl. Med. 17 (1990) 346-364 and Chem. Rev. 93 (1993) 1137-1156 described.

Suitable for SPECT are the complexes with the isotopes ¹¹¹ In and ^9m Tc.

Another imaging method with radioisotopes is positron emission tomography, the positron-emitting isotopes such. ⁴³ Sc, ⁴⁴ Sc, ⁵² Fe, ⁵⁵ Co, ⁶⁸ Ga, ⁶⁴ Cu, ⁸⁶ Y and ^94m Tc (Heiss, WD; Phelps, ME; Positron Emission Tomography of Brain, Springer Verlag Berlin, Heidelberg, New York 1983 ).

The compounds of the invention are surprisingly also for differentiation of benign and benign tumors in areas without a blood-brain barrier.

They are also characterized by the fact that they are completely eliminated from the body and thus are well tolerated.

Since the substances according to the invention accumulate in malignant tumors (no diffusion into healthy tissue, but high permeability of tumor vessels), they can also support the radiation therapy of malignant tumors. This differs from the corresponding diagnosis only by the amount and type of isotope used. The goal here is the destruction of tumor cells by high-energy short-wave radiation with the shortest possible range. For this purpose, interactions of the metals contained in the complexes (such as, for example, iron or gadolinium) with ionizing radiations (for example X-rays) or with neutron beams are utilized. This effect significantly increases the local radiation dose at the site where the metal complex is located (eg in tumors). In order to produce the same radiation dose in the malignant tissue, the application of such metal complexes can considerably reduce the radiation exposure for healthy tissue and thus avoid stressful side effects for the patients. The metal complex conjugates according to the invention are therefore also suitable as a radiosensitizing substance in the radiotherapy of malignant tumors (eg exploitation of Mössbauer effects or in neutron capture therapy). Suitable β-emitting ions are z. ⁴⁶ Sc, ⁴⁷ Sc, ⁴⁸ Sc, ⁷² Ga, ⁷³ Ga, ⁹⁰ Y, ⁶⁷ Cu, ¹⁰⁹ Pd, ¹¹¹ Ag, ¹⁴⁹ Pm, ¹⁵³ Sm, ¹⁶⁶ Ho, ¹⁷⁷ Lu, ¹⁸⁶ Re and ¹⁸⁸ Re, where ⁹⁰ Y , ¹⁷⁷ Lu, ⁷² Ga, ¹⁵³ Sm and ⁶⁷ Cu are preferred. Suitable low half-life having α-emitting ions are z. ²¹¹ At, ²¹¹ Bi, ²¹² Bi, ²¹³ , Bi and ²¹⁴ Bi, with ²¹² Bi being preferred. A suitable photon and electron emitting ion is ¹⁵⁸ Gd, which can be obtained from ¹⁵⁷ Gd by neutron capture.

If the agent according to the invention is for use in the method described by RL Mills et al. [Nature Vol. 336, (1988), p. 787], the central ion must be derived from a Mössbauer isotope such as ⁵⁷ Fe or ¹⁵¹ Eu.

In the case of in vivo application of the therapeutic agents according to the invention, these can together with a suitable carrier such. B. serum or physiological Saline and together with another protein such. B. Human serum albumin be administered. The dosage depends on the type of cellular disorder, the metal ion used and the type of imaging method.

The therapeutic agents according to the invention are administered parenterally, preferably i. v. applied.

Details of the applications of radiotherapeutics are z. In R.W. Kozak et al. TIBTEC, October 1986, 262 (see, inter alia, Bioconjugate Chem. 12 (2001) 7-34).

Overall, new complexing agents, metal complexes and metal complex salts have been achieved to synthesize the new possibilities in the diagnostic and therapeutic Open up medicine.

The following examples serve to explain the subject of the invention in more detail:

example 1 a) 2-tert-Butoxycarbonylamino-3- (4-nitro-phenyl) -propionic acid methyl ester

C

₁₅

H

₂₀

N

₂

O

₆

 (M = 324.34)

A suspension of 50 g (161.0 mmol) of Boc-NO ₂ -Phe (Bachem), 40.5 g (483 mmol) of sodium bicarbonate and 25.0 g (177 mmol) of methyl iodide in 600 ml of dimethylformamide was stirred for 4 days at room temperature. The suspension was filtered off with suction, the solid was washed with dichloromethane. The filtrate was concentrated on a rotary evaporator. The residue was taken up in water and extracted four times with ethyl acetate (150 ml each time). The combined organic phases were washed with 100 ml of 5% sodium thiosulfate solution, with 100 ml of 5% sodium bicarbonate solution, with 50 ml of saturated sodium chloride solution, with 100 ml of 10% citric acid and 50 ml of water. The organic phase was dried over sodium sulfate, filtered and concentrated on a rotary evaporator. This gave a crude yield of 46.5 g (143.5 mmol), which corresponds to 89.4%.
Re: C 55.55 H 6.22 N 8.64 O 29.60
Found: C 55.59 H 6.23 N 8.62 O 29.63 b) [tert-butyl 2-hydroxy-1- (4-nitro-benzyl) -ethyl] -carbamate C ₁₄ H ₂₀ N ₂ O ₅ (M = 296.32 )

8.9 g (27.4 mmol) of 1a were dissolved in 70 ml of tetrahydrofuran. 2.0 g (51.2 mmol) of sodium borohydride was added. There were slowly added dropwise 13 ml of methanol. The reaction solution was stirred overnight. The reaction solution was mixed with 2.8 ml of acetic acid and concentrated to dryness. The residue was taken up in water and extracted with ethyl acetate. The combined organic phases were washed twice with saturated sodium chloride solution. The organic phase was dried over sodium sulfate, filtered off and concentrated. Remaining water was removed with toluene by azeotropic distillation on a rotary evaporator. The desired crude product resulted in 79.0% yield (6.4 g, 21.6 mmol).
Calc .: C 56.75 H 6.80 N 9.45 O 27.00
Found: C 56.69 H 6.75 N 9.47 O 27.07 c) Methanesulfonic acid 2-tert-butoxycarbonylamino-3- (4-nitro-phenyl) -propyl ester C ₁₅ H ₂₂ N ₂ O ₇ S (M = 374.41)

6.40 g (21.6 mmol) of 1b were dissolved in 5 ml of dichloromethane and 0.33 g (3.24 mmol) of triethylamine. The solution was cooled to -5 ° C and slowly added with 0.27 g (2.38 mmol) of methanesulfonyl chloride, which had been diluted in a little dichloromethane. The suspension was stirred for 2 hours and poured onto stirred ice-water (about 50 ml). The phases were separated. The aqueous phase was extracted three times with dichloromethane (50 ml each). The combined organic phases were washed twice with dilute (5%) aqueous HCl solution, with water, with dilute (5%) sodium bicarbonate solution and with saturated sodium chloride solution. The organic phase was dried over sodium sulfate and concentrated on a rotary evaporator to dryness. There was 98.9% yield of crude product.
Re: C 48.12 H 5.92 N 7.48 O 29.91 S 8,56
Found: C 48.21 H 5.98 N 7.43 O 29.90 S 8.57 d) [2- (3-Amino-pentylamino) -1- (4-nitro-benzyl) -ethyl] -carbamic acid tert-butyl ester C ₁₉ H ₃₂ N ₄ O ₄ (M = 380.49)

10 g (26.71 mmol) of 1c were reacted with 27.85 g (267.1 mmol) of 1,3-diamino-pentane, 2.96 g (29.3 mmol) of triethylamine and 100 ml of tetrahydrofuran in analogy to Example 3a , There was 50.6% yield (5.14 g, 13.51 mmol) of the desired product. In addition, 3.75 g (9.87 mmol) of [2- (3-amino-1-ethyl-propylamino) -1- (4-nitro-benzyl) -ethyl] -carbamic acid tert-butyl ester were isolated.
Re: C 59.98 H 8.84 N 14.73 O 16.82
Found: C 59.90 H 8.86 N 14.75 O 16.77 e) N ¹ - [2-Amino-3- (4-nitro-phenyl) -propyl] -pentane-1,3-diamine C ₁₄ H ₂₄ N ₄ O ₂ ( M = 280.37)

3.65 g (9.59 mmol) of 1d were dissolved in 55 ml of dichloromethane and admixed with 16.24 g (142.46 mmol) of trifluoroacetic acid. The solution was stirred for 90 minutes at room temperature and concentrated on a rotary evaporator. It was added twice with dichloromethane and concentrated again. The crude product was mixed with 100 ml of 5% ammonia solution. The product was freeze-dried. The last described step was repeated. The desired product was found to be 2.39 g (8.54 mmol, 89%).
Re: C 59.98 H 8.63 N 19.98 O 11.41
Foundations: C 59.90 H 8.62 N 19.92 O 11.44 f) {[2 - {[3- (bis-tert-butoxycarbonylmethyl-amino) -pentyl] -tert-butoxycarbonylmethyl-amino} -1- (4-nitro-) Benzyl) ethyl] -tert-butoxycarbonylmethyl-amino} -acetic acid tert-butyl ester C ₄₄ H ₇₄ N ₄ O ₁₂ (M = 851.08)

To a solution of 5.27 g (18.8 mmol) of 1e in 246 ml of acetonitrile-water mixture (5: 1) were added 31.06 g (224.72 mmol) of potassium carbonate and 27.56 g (141.3 mmol ) Bromoacetic acid tert-butyl ester added. The reaction suspension was heated to 70 ° C and stirred for 24 hours. The suspension was concentrated on a rotary evaporator, treated with 350 ml of water and extracted three times with ethyl acetate. The organic phase was dried over sodium sulfate, filtered off and concentrated on a rotary evaporator to dryness. The crude product was purified by column chromatography (SiO ₂ , dichloromethane → dichloromethane: methanol 8: 1). The desired product was obtained in 71.0% yield (11.36 g, 13.35 mmol).
Calc .: C 62.10 H 8.76 N 6.58 O 22.56
Found: C 61.98 H 8.75 N 6.57 O 22.59 g) {[2 - {[3- (bis-carboxymethyl-amino) -pentyl] -carboxymethyl-amino} -1- (4-nitro-benzyl) -ethyl] - carboxymethylamino} -acetic acid C ₂₄ H ₃₄ N ₄ O ₁₂ (M = 570.55)

20.18 g (23.71 mmol) of 1f were initially charged in anisole. It was cooled to 0 ° C. 23 ml of trifluoroacetic acid were added. It was stirred at room temperature for 24 hours. The reaction solution was concentrated. The residue was taken up in water and extracted three times with diethyl ether. The aqueous phase was mixed with 100 ml of 5% ammonia solution and freeze-dried. The last described step was repeated. The desired product was obtained in 91% yield (12.31 g, 21.58 mmol).
Re: C 50.52 H 6.01 N 9.82 O 33.65
Found .: C 50.42 H 6.00 N 9.79 O 33.69

Example 2 a) N ³ - [2-Amino-3- (4-nitro-phenyl) -propyl] -pentane-1,3-diamine

C

₁₄

H

₂₄

N

₄

O

₂

 (M = 280.37)

2.92 g (7.67 mmol) of isolated by-product from 1d were dissolved in 40 ml of dichloromethane and treated with 12.99 g (114.0 mmol) of trifluoroacetic acid. The solution was stirred for 90 minutes at room temperature and concentrated on a rotary evaporator. It was added twice with dichloromethane and concentrated again. The crude product was mixed with 100 ml of 5% ammonia solution and freeze-dried. The last step was repeated. The desired product was obtained in 87% yield (1.87 g, 6.67 mmol).
Re: C 59.98 H 8.63 N 19.98 O 11.41
Found: C 59.89 H 8.60 N 19.93 O 11.39 b) {[ 2- {[3- (bis-tert-butoxycarbonylmethylamino) -1-ethyl-propyl] -tert-butoxycarbonylmethyl-amino} -1- (4- tert-butyl nitro-benzyl) -ethyl] -tert-butoxycarbonylmethyl-amino-acetic acid C ₄₄ H ₇₄ N ₄ O ₁₂ (M = 851.08)

To a solution of 6.18 g (7.14 mmol) of 2a in 246 ml of acetonitrile-water mixture (5: 1) was added 11.80 g (85.4 mmol) of potassium carbonate and 10.47 g (53.7 mmol ) Bromoacetic acid tert-butyl ester added. The reaction suspension was heated to 70 ° C and stirred for 24 hours. The suspension was concentrated on a rotary evaporator, treated with 350 ml of water and extracted three times with ethyl acetate. The organic phase was dried over sodium sulfate, filtered off and concentrated on a rotary evaporator to dryness. The crude product was purified by column chromatography (SiO ₂ , dichloromethane → dichloromethane: methanol 8: 1). The desired product was obtained in 73.0% yield (4.44 g, 5.21 mmol).
Calc .: C 62.10 H 8.76 N 6.58 O 22.56
Foundations: C 61.98 H 8.77 N 6.60 O 22.55 c) {[2 - {[3- (Bis-carboxymethyl-amino) -1-ethyl-propyl] -carboxymethyl-amino} -1- (4-nitrobenzyl) -ethyl] -carboxymethyl-amino} -acetic acid C ₂₄ H ₃₄ N ₄ O ₁₂ (M = 570.55)

1.35 g (1.58 mmol) of 2b were initially charged in anisole. It was cooled to 0 ° C. 1.53 ml of trifluoroacetic acid were added. It was stirred at room temperature for 24 hours. The reaction solution was concentrated. The residue was taken up in water and extracted three times with diethyl ether. The aqueous phase was concentrated with 100 ml of 5% ammonia solution and freeze-dried. The last mentioned step was repeated. The desired product was obtained in 88% yield (793 mg, 1.39 mmol).
Re: C 50.52 H 6.01 N 9.82 O 33.65
Found .: C 50.43 H 6.02 N 9.80 O 33.61

Example 3 a) N- [2- (3-amino-2,2-dimethyl-propylamino) -1- (4-nitro-benzyl) -ethyl] -2,2-dimethyl propionamide

C

₁₉

H

₃₂

N

₄

O

₃

 (M = 364.49)

To a solution of 5.0 g (13.4 mmol) of mesylate from Example 1c in 5 mL of tetrahydrofuran and 2.1 mL (14.7 mmol) of triethylamine were added 13.8 g (134 mmol) of 1,3-diamino-2 , 2-dimethylpropane added. The solution was heated to 50 ° C for four hours. The solution was concentrated on a rotary evaporator. The residue was taken up in water and extracted three times with ethyl acetate. The organic phase dried over sodium sulfate, filtered off and concentrated on a rotary evaporator. The residue was purified by column chromatography (SiO ₂ , dichloromethane → dichloromethane: methanol 1: 1 → methanol: ammonia (10%) 10: 1). Yield 73.2% (3.58 g, 9.80 mmol).
Re: C 62.61 H 8.85 N 15.37 O 13.17
Found: C 62.57 H 8.86 N 15.39 O 13.17 b) N ¹ - (3-Amino-2,2-dimethyl-propyl) -3- (4-nitrophenyl) -propane-1,2-diamine C ₁₄ H ₂₄ N ₄ O ₂ (M = 280.37)

5.23 g (13.75 mmol) of 3a were dissolved in 78 ml of dichloromethane and then admixed with 23.3 g (204.26 mmol) of trifluoroacetic acid. The solution was stirred for 90 minutes and concentrated. The residue was taken up in 100 ml of 5% ammonia solution and freeze-dried. The last step was repeated. This gave 8.21 g of product (13.2 mmol, 95.9%).
Re: C 59.98 H 8.63 N 19.98 O 11.41
Foundations: C 59.91 H 8.60 N 19.91 O 11.45 c) [(3 - {[2- (bis-tert-butoxycarbonylmethyl-amino) -3- (4-nitro-phenyl) -propyl] -tert-butoxycarbonylmethyl-amino} -2,2-dimethyl-propyl) -tert-butoxycarbonylmethyl-amino-acetic acid tert-butyl ester C ₄₄ H ₇₄ N ₄ O ₁₂ (M = 851.08)

8.02 g (12.9 mmol) of 3b were dissolved in 170 ml of acetonitrile-water mixture (5: 1) with 21.23 g (153.6 mmol) of potassium carbonate and 13.6 g (69.7 mmol) of bromoacetic acid tert-butyl ester. The reaction solution was heated at 70 ° C for 24 hours. The suspension was concentrated on a rotary evaporator. The residue was taken up in 300 ml of water and washed three times with ethyl acetate. The organic phase was dried with sodium sulfate, filtered off and concentrated to dryness on a rotary evaporator. The residue was purified by column chromatography (SiO ₂ , hexane-ethyl acetate 1: 1). This gave 9.79 g (11.5 mmol, 89.32%) of the title product.
Re: C 62.10 H 8.76 N 6.58 O 22.56
Found: C 62.19 H 8.74 N 6.57 O 22.59 d) [(3 - {[2- (Bis-carboxymethyl-amino) -3- (4-nitro-phenyl) -propyl] -carboxymethyl-amino} -2,2 -dimethyl-propyl) -carboxymethyl-amino] -acetic acid C ₂₄ H ₃₄ N ₄ O ₁₂ (M = 570.55)

5.30 g (6.23 mmol) of 3c were placed in 43 ml of anisole at -5 ° C. 6.15 ml (79.8 mmol) of trifluoroacetic acid were added. It was stirred overnight at room temperature. The reaction solution was concentrated. The residue was taken up in water and extracted three times with diethyl ether. The aqueous phase was mixed with 100 ml of 5% ammonia solution and freeze-dried. This gave 2.95 g (5.17 mmol) of the desired product. That corresponds to a yield of 83%.
Re: C 50.52 H 6.01 N 9.82 O 33.65
Found .: C 50.43 H 5.99 N 9.85 O 33.63

Example 4 a) 3-hydroxy-2-methyl-propionamide

C

₄

H

₉

NO (M = 103.12)

Analogous to: J. Amer. Chem. Soc .; 117; 9; (1995); 2479-2490. A solution of 30.0 g (225 mmol) of β-hydroxy-isobutyric acid methyl ester and 750 ml of a 9 M ammoniacal methanol solution was stirred in an airtight glass vessel for 7 days at 50 ° C. The solution was concentrated in vacuo. The residue was washed with cold diethyl ether (total 200 ml). There remained a white solid of 15.77 g (153 mmol, 68%).
Re: C 46.59 H 8.80 N 13.58 O 31.03
Found: C 46.63 H 8.83 N 13.55 O 31.06 b) 3-Amino-2-methyl-propan-1-ol C ₄ H ₁₁ NO (M = 89.14)

14.7 g (140 mmol) of 4a were suspended in 50 ml of tetrahydrofuran and mixed at 0 ° C with 400 ml (400 mmol) of 1M-borane-THF complex solution. The solution was heated at reflux for 4 hours and treated at 0 ° C with 70 ml of concentrated HCl solution. The solution was concentrated on a rotary evaporator. Diluted sodium hydroxide solution (140 g of sodium hydroxide in 200 ml of water) was added to the solution at 0 ° C. It was extracted four times with 100 ml of chloroform each time. The combined organic phases were dried with magnesium sulfate, filtered and concentrated. The residue was distilled in a water jet vacuum (92 ° C). The desired product was found to be 9.23 g (103.6 mmol, 74%).
Re: C 53.90 H 12.44 N 15.71 O 17.95
Found: C 53.80 H 12.42 N 15.68 O 17.98 c) tert-butyl (3-hydroxy-2-methyl-propyl) -carbamate C ₉ H ₁₉ NO ₃ (M = 189.25)

63.0 g (333 mmol) of 4b were dissolved in 240 ml of tetrahydrofuran and cooled to 0 ° C. At this temperature, 72.6 g (329.5 mmol) of di-tert-butyl dicarbonate ((Boc) ₂ O), dissolved in 95 ml of THF, were added dropwise. It was warmed to room temperature and stirred for one hour. The solution was concentrated on a rotary evaporator. The residue was taken up in 400 ml of diethyl ether and washed with 100 ml of 0.01N HCl, 100 ml of water and with 100 ml of sodium bicarbonate solution (5%). The organic phase was dried over sodium sulfate, filtered and concentrated on a rotary evaporator.
Calc .: C 57.12 H 10.12 N 7.40 O 25.36
Found: C 57.20 H 10.10 N 7.40 O 25.39 d) Methanesulfonic acid 3-tert-butoxycarbonylamino-2-methyl-propyl ester C ₁₀ H ₂₁ NO ₅ S (M = 267.34)

To 27.0 g (142.7 mmol) of 4c in 155 mL of dichloromethane was added 29.7 mL (214.1 mmol) of triethylamine. The solution was cooled to -5 ° C and treated with 11.7 ml (149.8 mmol) of methanesulfonyl chloride, which had been previously dissolved in 155 ml of dichloromethane. The suspension was stirred for 2 hours and treated with 400 ml of water. The phases were separated. The aqueous phase was extracted twice with 150 ml dichloromethane. The combined organic phases were washed twice with 200 ml of 0.1N HCl solution, once with 200 ml of 5% sodium bicarbonate solution and once with 100 ml of water. The organic phase was dried with sodium sulfate, filtered off and concentrated on a rotary evaporator. The residue was recrystallized in 0 ° C cold hexane. This gave 34.7 g (129.9 mmol) of the desired product; this corresponds to a yield of 91%.
Re: C 44.93 H 7.92 N 5.24 O 29.92 S 11.99
Found: C 44.90 H 7.89 N 5.24 O 29.90 S 12.01 e) (3-azido-2-methyl-propyl) -carbamic acid tert-butyl ester C ₉ H ₁₈ N ₄ O ₂ (M = 214.27)

71.6 g (268.6 mmol) of 4d were dissolved in 490 ml of DMSO and admixed with 21.0 g (322.3 mmol) of sodium azide. The reaction solution was stirred for 24 hours at 40-45 ° C. The mixture was cooled to 25 ° C and treated with 500 ml of water. The solution was extracted five times with 250 ml of dichloromethane. The combined organic phases were washed twice with 150 ml of saturated sodium chloride solution. The organic phase was dried with sodium sulfate, filtered off and concentrated on a rotary evaporator. The residue was purified by column chromatography (SiO ₂ , hexane → hexane-ethyl acetate 1: 1). After purification, there were 43.7 g (204 mmol) of product; this corresponds to a yield of 76%.
Re: C 50.45 H 8.47 N 26.15 O 14.93
Found: C 50.51 H 8.46 N 26.12 O 14.95 f) tert-butyl (3-amino-2-methyl-propyl) -carbamate C ₉ H ₂₀ N ₂ O ₂ (M = 188.27)

30.8 g (143.8 mmol) of 4e were dissolved in 412 ml of ethyl acetate and mixed with 4.5 g of Pd / C (10%). The reaction solution was stirred at 25 ° C under a hydrogen atmosphere for 24 hours. The solution was filtered and concentrated on a rotary evaporator. The residue was purified by column chromatography (SiO ₂ , dichloromethane → dichloromethane: methanol 1: 1). Yield 24.28 g (129.0 mmol, 89.7%).
Calc .: C 57.42 H 10.71 N 14.88 O 17.00
Found: C 57.45 H 10.73 N 14.90 O 16.99 g) tert-butyl {3- [2-tert-butoxycarbonylamino-3- (4-nitro-phenyl) -propionylamino] -2-methyl-propyl} -carbamate C ₂₃ H ₃₆ N ₄ O ₇ (M = 480.56)

21.7 g (115 mmol) of 4f were dissolved in 600 ml of dichloromethane / water (1: 1) and washed with 36.0 g (115 mmol) of Boc-protected nitrophenylalanine and 17.6 g (115 mmol) of 1-hydroxybenzotriazole - H ₂ O (HOBT) offset. The solution was cooled to about -5 ° C. There were added 24.0 g (127 mmol) of 1- (dimethylaminopropyl) -3-ethylcarbodiimide (EDCI) and stirred for 7 hours and a further three days at 25 ° C. The phases were separated. The aqueous phase was extracted twice with dichloromethane. The combined organic phases were washed twice with 150 ml of saturated sodium bicarbonate solution and a little water. The organic phase was dried with sodium sulfate, filtered off and concentrated on a rotary evaporator. The residue was pulverized in a mortar and washed with cold hexane. It was dried on the oil pump. Yield: 35.1 g (73.1 mmol, 63.6%).
Re: C 57.49 H 7.55 N 11.66 O 23.30
Found: C 57.46 H 7.55 N 11.67 O 23.32 h) 2-Amino-N- (3-amino-2-methyl-propyl) -3- (4-nitrophenyl) -propionamide C ₁₃ H ₂₀ N ₄ O ₃ (M = 280.33)

10.3 g (21.4 mmol) of 4 g were suspended in 120 ml of dry dichloromethane. 24.5 ml (318 mmol) of trifluoroacetic acid were added dropwise and the mixture was stirred for 1 hour. The mixture was concentrated on a rotary evaporator, treated with 100 ml of dichloromethane and concentrated again. The residue was washed with diethyl ether and dried at 40 ° C on the oil pump. 100 ml of ammonia solution (5%) was added. It was freeze-dried. The desired product was found to be 5.55 g (19.80 mmol, 92.5%).
Re: C 55.70 H 7.19 N 19.99 O 17.12
Found: C 55.39 H 7.21 N 19.89 O 17.19 i) (3-Amino-2-methyl-propyl) - [2-amino-3- (4-nitrophenyl) -propyl] -amine di-hydrochloride C ₁₃ H ₂₂ N ₄ O ₂ (M = 252.32)

9.25 g (18.2 mmol) of 4h were added in 130 ml of abs. THF solved. 128.5 ml (128.5 mmol) borane-THF complex (1 molar) were added dropwise at 0 ° C. within 30 min. The solution was warmed to room temperature and heated at reflux for 5 hours. The solution was cooled to 0 ° C, added with 35 ml of methanol, stirred for 2 hours and concentrated. The residue was dissolved in 200 ml of ethanol, cooled in an ice bath and mixed with hydrogen chloride gas. The mixture was concentrated and taken up in diethyl ether. The solid was filtered off with suction, rinsed with diethyl ether and dried in vacuo. This gave 5.49 g (14.6 mmol, 80.3%) of the desired product.
Calc .: C 41.56 H 6.71 N 14.91 O 8.52 Cl 28.31
Found: C 41.59 H 6.76 N 14.92 O 8.54 Cl 28.26 k) [(3 - {[2- (bis-tert-butoxycarbonylmethyl-amino) -3- (4-nitro-phenyl) -propyl] -tert-butoxycarbonylmethyl- amino-2-methyl-propyl-tert-butoxycarbonylmethyl-amino] -acetic acid tert-butyl ester C ₄₃ H ₇₂ N ₄ O ₁₂ (M = 837.06)

10.37 g (27.6 mmol) of 41 were dissolved in 300 ml of acetonitrile-water (5: 1) and treated with 45.5 g (329.2 mmol) of potassium carbonate. There were added 29.1 g (149.4 mmol) of tert-butyl bromoacetate. The reaction was stirred at 70 ° C for 24 hours. The reaction solution was mixed with 500 ml of water and extracted three times with 150 ml of ethyl acetate. The organic phase was dried with sodium sulfate, filtered and concentrated on a rotary evaporator. The residue was purified by column chromatography (SiO ₂ , dichloromethane → dichloromethane: methanol 8: 1). The desired product was obtained in 73.0% yield (16.87 g, 20.1 mmol).
Re: C 61.70 H 8.67 N 6.69 O 22.94
Found: C 61.67 H 8.66 N 6.71 O 22.91 1) [(3 - {[2- (Bis-carboxymethyl-amino) -3- (4-nitro-phenyl) -propyl] -carboxymethyl-amino} -2-methyl -propyl) -carboxymethyl-amino] -acetic acid C ₂₃ H ₃₂ N ₄ O ₁₂ (M = 556.52)

38.76 g (46.3 mmol) of 4k were initially charged in 318 ml of anisole and cooled to -5 ° C. 458 ml of trifluoroacetic acid were added. It was stirred for 3 hours at 0 ° C. It was warmed to room temperature and stirred for 24 hours. The reaction solution was concentrated on a rotary evaporator. The residue was taken up in 250 ml of water and extracted three times with diethyl ether. The water phase was concentrated. Methanol / ammonia (5%) was added and concentrated again. Then dissolved in water and freeze-dried. The desired product was obtained in 98.3% yield (40.9 g, 45.5 mmol).
Calc .: C 49.64 H 5.82 N 10.07 O 34.50
Found .: C 49.53 H 5.81 N 10.01 O 34.53

Example 5 a) diethyl 2-benzyl malonic acid

C

₁₄

H

₁₈

O

₄

 (M = 250.29)

The synthesis of the desired product is known in the literature (Synthesis, 12, 2000, 1749-1755).
Calc .: C 67.18 H 7.25 O 25.57
Found: C 67.15 H 7.26 O 25.54 b) 2-Benzyl-malonic acid amide C ₁₀ H ₁₂ N ₂ O ₂ (M = 192.22)

The synthesis of the desired product was carried out analogously to Example 15b.
Re: C 62.49 H 6.29 O 16.65 N 14.57
Found: C 62.59 H 6.30 O 16.64 N 14.59 c) 2-Benzyl-propane-1,3-diamine C ₁₀ H ₁₆ N ₂ (M = 164.25)

The reduction to the desired product was carried out analogously to Example 15c.
Re: C 73.13 H 9.82 N 17.06
Found: C 73.09 H 9.85 N 17.09 d) [(2-Benzyl-3 - {[2- (bis-carboxymethyl-amino) -3- (4-nitro-phenyl) -propyl] -carboxy-methyl-amino} -propyl ) -carboxymethyl-amino] -acetic acid C ₂₉ H ₃₆ N ₄ O ₁₂ (M = 632.62)

The synthesis of the desired product and its precursors were carried out analogously to Example 15c.
Re: C 55.06 H 5.74 N 8.86 O 30.35
Found: C 55.09 H 5.72 N 8.84 O 30.32

Example 6 Gd complex of the compound according to Example 3

gDNA

₂

C

₂₄

H

₂₉

N

₄

O

₁₂

 (M = 768.74)

142.6 mg (0.25 mmol) of 3d were suspended in 4 ml of distilled water, heated to 80 ° C and dissolved. 45.3 mg (0.125 mmol) of Gd ₂ O _{3 were} added in portions. The suspension was heated to 80 ° C and stirred for one hour. The solution was cooled to room temperature and adjusted to pH = 7 with sodium hydroxide solution (1 M). The water was removed by lyophilization. The desired was found to be 192.2 mg (0.25 mmol, 99.8%).
Calc .: C 37.50 H 3.80 N 7.29 O 24.97 Gd 13.87 Na 5.98
Result: C 37.49 H 3.77 N 7.31 O 24.99 Gd 13.89 Na 6.01

Example 7 ({3 - [(2- (bis-carboxymethyl-amino) -3- {4- [3- (2,5-dioxo-2,5-dihydro-pyrrol-1-yl) - propionylamino] -phenyl} -propyl) -carboxymethyl-amino] -2,2-dimethyl-propyl} - carboxymethyl-amino) -acetic acid

C

₃₁

H

₄₁

N

₅

O

₁₃

 (M = 691.69)

270.3 mg (0.5 mmol) of aniline derivative 8 and 328 ml (4.54 mmol) of N-methylmorpholine were dissolved in 2.5 ml of dimethylsulfoxide and charged with 134.16 mg (0.61 mmol) of maleimide, MPHS (Fluka) active ester. , offset. The reaction solution was heated to give a homogeneous solution. The mixture was stirred for 40 min and concentrated in vacuo. The residue was purified by RP-HPLC. 180 mg (0.26 mmol, 52%) of the desired product were obtained.
Re: C 53.83 H 5.97 N 10.13 O 30.07
App .: C 53.74 H 6.00 N 10.08 O 30.09

Example 8 [(3 - {[3- (4-Amino-phenyl) -2- (bis-carboxymethyl-amino) -propyl] -carboxymethyl-amino} - 2,2-dimethyl-propyl) -carboxymethyl-amino] -acetic acid

C

₂₄

H

₃₆

N

₄

O

₁₀

 (M = 540.57)

998 mg (1.75 mmol) of 3d and 0.6 g of palladium on carbon (10%) were dissolved in 30 ml of methanol-water (4: 1) and hydrogenated under hydrogen atmosphere (normal pressure) at room temperature until the calculated amount of hydrogen ( 39.2 ml). It was filtered and washed with methanol. It was concentrated on a rotary evaporator, suspended in toluene and concentrated again. The desired product was obtained with 746 mg (1.38 mmol, 78.7% yield).
Re: C 53.33 H 6.71 N 10.36 O 29.60
Found .: C 53.41 H 6.74 N 10.42 O 29.61

Example 9 [(3 - {[2- (bis-carboxymethyl-amino) -3- (4-isothiocyanato-phenyl) -propyl] - carboxymethyl-amino} -2,2-dimethyl-propyl) -carboxymethyl-amino] -acetic acid

C

₂₅

H

₃₄

N

₄

O

₁₀

S (M = 582.63)

811 mg (1.5 mmol) of the product from Example 8 and 969 mg (9.14 mmol) of sodium carbonate were dissolved in 35 ml of dist. Dissolved water and 70 ml of chloroform. To this 2-phase system was added 132.8 ml (1.74 mmol) of thiophosgene. The solution was stirred for 3 hours. The solution was concentrated, taken up in 0.1 ml of dilute acetic acid (1%) and purified by RP-HPLC (25: 74: 1 acetonitrile / water / acetic acid). The desired product was obtained in 64.2% yield (561 mg, 963 mmol).
Calc .: C 51.54 H 5.88 N 9.62 O 27.46 S 5.50
Gef .: C 51.59 H 5.88; N 9.64 O 27.50 S 5.48

Example 10 {[3 - ({2- (bis-carboxymethyl-amino) -3- [4- (2-bromo-acetylamino) -phenyl] -propyl} - carboxymethyl-amino) -2,2-dimethyl-propyl] -carboxymethyl-amino} -acetic acid

C

₂₆

H

₃₇

BrN

₄

O

₁₁

 (M = 661.50)

81.1 mg (0.15 mmol) of product from Example 8 were dissolved in 3 ml of ethanol, 3 ml of dist. Dissolved water and 3 ml of saturated sodium bicarbonate and treated with 0.56 g (2.18 mmol) of bromoacetic anhydride. The addition of solid sodium bicarbonate kept the pH at 8.5. The reaction solution was stirred for 1 hour and concentrated. The residue was filtered through cotton wool, rinsed with ethanol, concentrated and purified by RP-HPLC. There were 79.4 mg (0.12 mmol) of product. That corresponds to a yield of 80%.
Re: C 47.21 H 5.64 N 8.47 O 26.60 Br 12.08
Gef .: C 47.11 H 5.68 N 8.49 O 26.54 Br 12.00

Example 11 {[3 - ({2- (bis-carboxymethyl-amino) -3- [4- (2-iodo-acetylamino) -phenyl] -propyl} - carboxymethyl-amino) -2,2-dimethyl-propyl] -carboxymethyl-amino} -acetic acid

C

₂₆

H

₃₇

IN

₄

O

₁₁

 (M = 708.50)

81.1 mg (0.15 mmol) of product from Example 8 were dissolved in 3 ml of ethanol, 3 ml of dist. Dissolved in water and 3 ml of saturated sodium bicarbonate and treated with 771 mg (2.18 mmol) of iodoacetic anhydride. The addition of solid sodium bicarbonate kept the pH at 8.5. The reaction solution was stirred for 1 hour and concentrated. The residue was filtered through cotton wool, rinsed with ethanol, concentrated and purified by RP-HPLC. This gave a yield of 69% (73.3 mg, 0.104 mmol) of the desired product.
Re: C 44.08 H 5.26 N 7.91 O 24.84 I 17.91
Found .: C 44.04 H 5.29 N 7.89 O 24.75 I 17.99

Example 12 [(3 - {[2- (bis-carboxymethyl-amino) -3- (4-thiosemicarbazido-phenyl) -propyl] - carboxymethyl-amino} -2,2-dimethyl-propyl) -carboxymethyl-amino] -acetic acid

C

₂₅

H

₃₈

N

₆

O

₁₀

S (M = 614.67)

The desired product was obtained from the compound of Example 8, analogous to the procedure in Collect. Czech. Chem. Commun., 57, 3, (1992), 656-659.
Re: C 48.85; H 6.23; N 13.67; O 26.03; S 5.22
Found: C 48.77; H 6.25; N 13.62; O 26.06; S 5.25

Example 13 [(3 - {[3- (4-acetylamino-phenyl) -2- (bis-carboxymethyl-amino) -propyl] -carboxymethyl- amino} -2,2-dimethyl-propyl) -carboxymethyl-amino] -acetic acid

C

₂₆

H

₃₈

N

₄

O

₁₁

 (M = 582.60)

Analogous to J. Amer. Chem. Soc., 120; 12; 1998; 2768-2779:
81.1 mg (0.15 mmol) of the product from Example 8 were dissolved in 1.5 ml of acetonitrile-H ₂ O (9: 1) was cooled to 0 ° C and treated with 38.3 mg (375 mmol) of acetic anhydride. The solution was stirred at room temperature for 4 hours. It was filtered and concentrated in vacuo.
Calc .: C 53.60 H 6.57 N 9.62 O 30.21
Found .: C 53.49 H 6.54 N 9.64 O 30.24

Example 14 a) 2,3-dimethyl-succinonitrile

C

₆

H

_8th

N

₂

 (M = 108.14)

The synthesis of 2,3-dimethyl-succinonitrile was carried out according to a procedure of Whiteley and Marianelli (Synthesis (1978), 392-394) from 8.7 g (149.8 mmol) of acetone, 16 ml (150.4 mmol) of ethyl cyanoacetate and 10 g (154 mmol) of potassium cyanide. The crude product was purified by distillation in vacuo. Yield 7.0 g (64.7 mmol, 61%).
Re: C 66.64 H 7.46 N 25.90
Found: C 66.60 H 7.44 N 25.92 b) 2,3-Dimethyl-butane-1,4-diamine C ₆ H ₁₆ N ₂ (M = 116.21)

The synthesis of 2,3-dimethyl-butane-1,4-diamine was carried out according to a protocol of Aizencang et al. (J. Med. Chem. 38; 21; 1995; 4337-4341): 10.81 mg (100 mmol) of 2,3-dimethyl-succinonitrile were reacted with saturated diborane-THF solution. This gave 8.02 g (69 mmol, 69%) of the desired product as a colorless liquid.
C 62.02 H 13.88 N 24.11
C 62.19 H 13.90 N 24.12 c) [(4 - {[2- (bis-carboxymethyl-amino) -3- (4-nitro-phenyl) -propyl] -carboxymethyl-amino} -2,3-dimethyl-butyl) carboxymethyl-amino] -acetic acid C ₂₅ H ₃₆ N ₄ O ₁₂ (M = 584.58)

The synthesis of the desired product and the associated precursors was carried out analogously to Example 3.
Re: C 51.37 H 6.21 N 9.58 O 32.84
Found .: C 51.34 H 6.23 N 9.61 O 32.80

Example 15 a) cyclopentane-1,1-dicarboxylic acid diethyl ester

C

₁₁

H

₁₈

O

₄

 (M = 214.26)

The synthesis of the substance was prepared from 1,4-dibromobutane and malonic acid diester according to a protocol from J. Amer. Chem. Soc., 109; 22; , 1987; 6825-6836
Re: C 61.66 H 8.47 O 29.87
Found: C 61.69 H 8.46 O 29.82 b) cyclopentane-1,1-dicarboxylic acid diamide C ₇ H ₁₂ N ₂ O ₂ (M = 156.18)

A solution of 21.4 g (100 mmol) of 15a and 500 ml of a 9M ammoniacal methanol solution was stirred in an airtight glass jar for 7 days at 50 ° C. The solution was concentrated in vacuo. The residue was washed with cold diethyl ether (total 200 ml). There remained a white solid of 10.0 g (64 mmol, 64%).
Re: C 53.83 H 7.74 N 17.97 O 20.49
Found: C 53.80 H 7.71 N 18.00 O 20.52 c) C- (1-Aminomethyl-cyclopentyl) -methylamine C ₇ H ₁₆ N ₂ (M = 128.22)

9.9 g (63.4 mmol) of 15b were suspended in 50 ml of tetrahydrofuran. It was at 0 ° C Added 400 ml (400 mmol) of 1M borane-THF complex solution. The solution was for 4 Heated for hours and mixed at 0 ° C with 70 ml of concentrated HCl solution.

The solution was concentrated on a rotary evaporator. Diluted sodium hydroxide solution (140 g of sodium hydroxide in 200 ml of water) was added to the solution at 0 ° C. It was extracted four times with 100 ml of chloroform each time. The combined organic phases were dried with magnesium sulfate, filtered and concentrated. The residue was distilled in vacuo. The desired product was found to be 5.85 g (45.64 mmol, 72%).
Re: C 65.57 H 12.58 N 21.85
Foundations: C 65.49 H 12.59 N 21.87 d) {[ 1- ({[2- (Bis-carboxymethyl-amino) -3- (4-nitro-phenyl) -propyl] -carboxymethyl-amino} -methyl) -cyclopentylmethyl ] -carboxymethyl-amino} -acetic acid C ₂₆ H ₃₆ N ₄ O ₁₂ (M = 596.59)

The synthesis of the desired product and the associated precursors was carried out analogously to Example 3 with the diamine 15c.
Re: C 52.35 H 6.08 N 9.39 O 32.18
Found .: C 52.43 H 6.09 N 9.40 O 32.21

Example 16 trans - [(4 - {[2- (bis-carboxymethyl-amino) -3- (4-nitro-phenyl) -propyl] -carboxymethyl- amino} -cyclohexyl) -carboxymethyl-amino] -acetic acid

C

₂₅

H

₃₄

N

₄

O

₁₂

 (M = 584.58)

The synthesis of the desired product and the associated precursors was carried out analogously to Example 3 with trans-1,4-diaminocyclohexane.
Re: C 51.37 H 6.21 N 9.58 O 32.84
Found .: C 51.28 H 6.23 N 9.60 O 32.88

Example 17 a) cis-2-aminomethylcyclohexylamine

C

₆

H

₁₄

N

₂

 (M = 114.19)

The compound has been prepared as described in the literature (Tetrahedron, 44, 5, 1988, 1465-1476).
Re: C 65.57 H 12.58 N 21.85
Foundations: C 65.51 H 12.61 N 21.87 b) {[2 - {[2- (Bis-carboxymethyl-amino) -cyclohexylmethyl] -carboxymethyl-amino} -1- (4-nitro-benzyl) -ethyl] -carboxymethyl amino} -acetic acid C ₂₆ H ₃₆ N ₄ O ₁₂ (M = 596.59)

The synthesis of the desired product and the associated precursors was carried out analogously to Example 3 with the diamine 17a.
Re: C 52.35 H 6.08 N 9.39 O 32.18
Found .: C 52.30 H 6.09 N 9.36 O 32.20

Example 18 Antibody conjugate of {[3 - ({2- (bis-carboxymethylamino) -3- [4- (2-bromo- acetylamino) -phenyl] -propyl} -carboxymethyl-amino) -2,2-dimethyl-propyl] - carboxymethyl-amino} acetic acid

200 μg of an antibody with freely accessible thiol groups (eg HuM195 (see Michael R. McDevitt, J. Nuc. Med. 40, 1999, 1722; commercially available from Protein Design Labs Inc., Mountainview, CA, USA) - the antibody has no freely available thiol groups, these can be generated by the use of 2-iminothiolane HCl (eg EP 0 607 222 B1)) were diluted in 1.2 ml borate buffer (50 mM, pH 8.5) containing 238 μg (240 nmol). Product from Example 10, dissolved in 50 μl of borate buffer (see above), added and 3 hours at 37 ° C. touched. It was isolated on a NAP-5 column (Amersham Pharmacia Biotech AB, Sephadex G 25, Mobile phase: PBS).

Example 19 Indium 111-labeled antibody conjugate of {[3 - ({2- (bis-carboxymethyl-amino) -3- [4- (2-bromo-acetylamino) -phenyl] -propyl} -carboxymethyl-amino) -2,2-dimethyl- propyl] amino} acetic acid -carboxymethyl-

200 μg of an antibody with thiol groups which are freely available (eg HuM195 (see Michael R. McDevitt, J. Nuc. Med. 40, 1999, 1722, commercially available from Protein Design Labs Inc., Mountainview, CA, USA), if the antibody does not have freely accessible thiol groups, these can be generated by the use of 2-iminothiolane HCl (eg EP 0 607 222 B1)) were diluted in 1.2 ml borate buffer (50 mM, pH 8.5), with 238 ug (240 nmol) product of Example 10, dissolved in 50 ul borate buffer (see above), and stirred at 37 ° C for 3 hours. The borate buffer solution was replaced with an acetate buffer by placing the sample solution in the Slide-A-Lyzer 10000, Pierce MWCO (dialysis method) three times for 1 h each against 200 ml of NaOAc buffer 0.1 M (pH 6.0) has been. Finally, overnight against 400 ml of NaOAc buffer 0.1 M (pH 6) was provided. The solution was added with 80 μl (0.05 M HCl) [ ¹¹¹ In] InCl ₃ (27.88 MBq) and stirred for 30 min at room temperature. It was purified on a NAP-5 column (Amersham Pharmacia Biotech AB, Sephadex G-25, Mobile phase: PBS).

Example 20 Yttrium 90-labeled antibody conjugate of {[3 - ({2- (bis-carboxymethyl-amino) -3- [4-] (2-bromo-acetylamino) -phenyl] -propyl} -carboxymethyl-amino) -2,2-dimethyl-propyl] - carboxymethyl-amino} acetic acid

200 μg of an antibody with thiol groups which are freely available (eg HuM195 (see Michael R. McDevitt, J. Nuc. Med. 40, 1999, 1722, commercially available from Protein Design Labs Inc., Mountainview, CA, USA), if the antibody does not have freely accessible thiol groups, these can be generated by the use of 2-iminothiolane HCl (eg EP 0 607 222 B1)) were diluted in 1.2 ml borate buffer (50 mM, pH 8.5), with 238 ug (240 nmol) product of Example 10, dissolved in 50 ul borate buffer (see above), and stirred at 37 ° C for 3 hours. The borate buffer solution was replaced with an acetate buffer by diluting the sample solution three times for 1 h in the Slide-A-Lyzer 10000, Pierce, MWCO (dialysis method) against 200 ml NaOAc buffer 0.1 M (pH 6.0). was asked. Finally, overnight against 400 ml of NaOAc buffer 0.1 M (pH 6) was provided. The solution was added with 50 MBq of [ ⁹⁰ Y] YCl ₃ and stirred for 30 minutes at room temperature. It was purified on a NAP-5 column (Amersham Pharmacia Biotech AB, Sephadex G-25, Mobile phase: PBS).

Claims (19)

1. Compounds of the formula I


wherein
Z is a hydrogen atom or a metal ion equivalent,
A is a radical of the formula


stands, in which the with


labeled positions α and β are attached to any of the adjacent nitrogen atoms, R ^{1 is} a nitro group or a group capable of reacting with a biomolecule, and
B is a radical of the formula


in which n is 0 or 1 and R ² , R ³ , R ⁴ , R ⁵ , R ⁶ , R ⁷ , R ⁸ and R ^{9 are} independently selected from a hydrogen atom, a straight-chain or branched, saturated or unsaturated C _{1 -6-} alkyl group which may be optionally substituted with 1 or 2 hydroxy groups and / or may contain 1 or 2 oxygen atoms, and an aralkyl group, the aryl group may optionally be substituted by an alkyl or alkoxy group, wherein two of the radicals R ² , R ³ , R ⁴ , R ⁵ , R ⁶ , R ⁷ , R ⁸ and R ^{9 may be} part of a 5- or 6-membered ring, with the proviso that at least one and at most four of the radicals R ² , R ³ , R ⁴ , R ⁵ , R ⁶ , R ⁷ , R ⁸ and R ^{9 are} not hydrogen atoms, and their salts. 2. Compounds according to claim 1, wherein the radical A is bonded via the position α to the (ZOOC-CH ₂ ) ₂ -N-radical. 3. Compounds according to any of the preceding claims, wherein R ^{1 is} selected from the group consisting of nitro, amino, isocyanate, isothiocyanate, hydrazine, semicarbazide, thiosemicarbazide, chloroacetamide, bromoacetamide, iodoacetamide, acylamino, maleimide, maleimidacylamino, activated esters, mixed anhydrides , Azide, hydroxide, sulfonyl chloride and carbodiimide. 4. Compounds according to one of the preceding claims, wherein 1 or 2 of the radicals R ² , R ³ , R ⁴ , R ⁵ , R ⁶ , R ⁷ , R ⁸ and R ^{9 are} selected from the group consisting of methyl, ethyl and benzyl and the other of these radicals are hydrogen atoms. 5. Compounds according to any one of the preceding claims, wherein the radicals R ² , R ³ , R ⁴ , R ⁵ , R ⁶ , R ⁷ , R ⁸ and R ⁹ are selected such that B is symmetrical. 6. Compounds according to any one of claims 1-3, wherein B is selected from the group consisting of -CH ₂ -CH ₂ -CH (CH ₂ -CH ₃ ) -, -CH (CH ₂ -CH ₃ ) -CH ₂ - CH ₂ -, -CH ₂ -C (CH ₃ ) ₂ - CH ₂ -, -CH ₂ -CH (CH ₃ ) -CH ₂ -, -CH ₂ -CH (CH ₂ -phenyl) -CH ₂ -, - CH ₂ -CH (CH ₃ ) -CH (CH ₃ ) -CH ₂ -,


7. Compounds according to one of the preceding claims, wherein n = 0. 8. Compounds according to any one of the preceding claims, wherein at least two Z represents a metal ion equivalent of a paramagnetic element of Ordinal numbers 21-29, 42, 44 and 58-70 stand. 9. Compounds according to any one of claims 1-7, wherein at least two of the radicals Z for a metal ion equivalent of a radioactive element of atomic numbers 26, 27, 29, 31, 32, 37-39, 43, 46, 47, 49, 61, 62, 64, 67, 70, 71, 75, 77, 82 and 83. 10. Conjugates of the general formula II


wherein Z and B are as defined in claim 1 and A 'is a radical of the formula


stands, in which the with


labeled positions α and β are bound to any of the adjacent nitrogen atoms and Bio stands for the rest of a biomolecule, which is bound via the radical R ^{1 'of} a reactive group to the phenylene ring, and their salts. 11. Conjugates according to claim 10, wherein the biomolecule is selected from the group consisting of biopolymers, proteins, synthetically modified biopolymers, Carbohydrates, antibodies, DNA and RNA fragments, β-amino acids, vector amines for introduction into the cell, biogenic amines, pharmaceuticals, oncological Preparations, synthetic polymers directed to a biological target, Steroids, prostaglandins, taxol and its derivatives, endothelins, alkaloids, Folic acid and its derivatives, bioactive lipids, fats, fatty acid esters, synthetic modified mono-, di- and triglycerides, liposomes that derivatized at the surface are micelles of natural fatty acids or of perfluoroalkyl compounds, Porphyrins, texaphrins, expanded porphyrins, cytochromes, inhibitors, Neuramidases, neuropeptides, immunomodulators, endoglycosidases, substrates that by the enzymes calmodulin kinase, casein kinase II, glutathione-S-transferase, Heparinase, matrix methalloprotheases, β-insulin receptor kinase, UDP-galactose 4- Epimerase, fucosidases, G proteins, galactosidases, glycosidases, glycosyltransferases and xylosidase, antibiotics, vitamins and vitamin analogues, hormones, DNA intercalators, nucleosides, nucleotides, lectins, vitamin B12, Lewis-X and Relatives, psoralenes, dientenrienantibiotika, Carbacyclinen, VEGF, somatostatin and its derivatives, biotin derivatives, antihormones, tumor specific proteins and Synthetics, dendrimers and cascade polymers, as well as their derivatives, polymers, the accumulate in acidic or basic areas of the body, myoglobins, Apomyoglobins, neurotransmitter peptides, tumor necrosis factors, peptides found in accumulate inflamed tissues, blood pool reagents, anions and cations. Transporter proteins, polyesters, polyamides and polyphosphates. 12. Conjugates according to any one of claims 10 or 11, wherein at least two of the radicals Z is a metal ion equivalent of a paramagnetic element of atomic numbers 21-29, 42, 44 and 58-70. 13. Conjugates according to any one of claims 10 or 11, wherein at least two of the radicals Z for a metal ion equivalent of a radioactive element of atomic numbers 26, 27, 29, 31, 32, 37-39, 43, 46, 47, 49, 61, 62, 64, 67, 70, 71, 75, 77, 82 and 83 stand. 14. Use of a compound according to any one of claims 1-9 for the preparation of a Conjugate with a biomolecule. 15. Pharmaceutical agent containing at least one physiologically acceptable A compound according to any one of claims 8 or 9 or at least one physiological compatible conjugate according to one of claims 12 or 13, optionally with the in the galenics usual additives. 16. Use of a compound according to any one of claims 1-8 or a conjugate according to any one of claims 10-12 for the preparation of means for NMR diagnosis. 17. Use of a compound according to any one of claims 1-7 or 9 or one A conjugate according to any one of claims 10, 11 or 13 for the preparation of agents for the Radiodiagnosis or radiotherapy. 18. Kit for the preparation of radiopharmaceuticals, comprising a compound according to a of claims 1-7 or a conjugate according to any one of claims 10 or 11, wherein Z Is hydrogen, and a compound of a radioactive element of atomic numbers 26, 27, 29, 31, 32, 37-39, 43, 46, 47, 49, 61, 62, 64, 67, 70, 71, 75, 77, 82 and 83. 19. A process for the preparation of a compound according to claim 1 or a conjugate according to claim 10, wherein a compound of formula III

H ₂ NA-NH-B-NH ₂ III

wherein A and B are as defined in claim 1, with a compound of formula IV

Nu-CH ₂ -COOZ 'IV

where Nu is a nucleofuge and Z 'is a hydrogen atom, a metal ion equivalent or a protecting group for carbonyl, then the resulting compound is optionally reacted with a biomolecule, the radical R ¹ , when it is nitro, first in a group which can react with a biomolecule, is converted, and thereafter, after removal of any protective groups still present and in a conventional manner if desired with at least one metal oxide or metal salt is reacted and optionally then in the complexes thus obtained acidic hydrogen atoms are completely or partially substituted by cations of inorganic and / or organic bases, amino acids or amino acid amides.