CZ398A3 - Complexes of cascade polymers, process of their preparation and pharmaceutical composition containing thereof - Google Patents

Abstract

Described are new cascade polymer complexes, compositions containing them and use of the complexes in diagnosis and therapy, particularly for magnetic resonance imaging and computer tomography imaging.

Description

Field of technology

The invention relates to new complexes of cascade polymers, means containing these compounds, the use of complexes in diagnostics and therapy, as well as the method of production of these valuable compounds and means.

Current state of the art

Sometimes clinically used contrast agents for modern imaging procedures, namely nuclear magnetic resonance imaging (MRI) and computed tomography (CT) [Magnevist , R R R

For Hance , Ultravist and Omniscan ] they scatter throughout the extracellular space of the body (intravasal space and interstitium). This space of scattering comprises about 20% of the body volume.

Extracellular MRI-contrast means have been used clinically with success first in the diagnosis of brain and spinal disease processes, as a completely special situation occurs here with regard to the regional space of dispersion. In the brain and spinal cord, extracellular substances cannot leave the healthy tissue based on the blood-brain barrier within the vascular space. During disease processes with disorders of this blood-brain barrier (for example, malignant tumors, inflammation, myelopathic diseases, etc.), areas with increased permeability (permeability) of vessels then arise inside the brain »· · · · « « · · A • · · * » A Α * ·« · • * · A Α · ···· • 4 · * * * · · »»· ♦ · ··· »«« »·« • · ···· · · 1 A Τ A Α «I for these extracellular contrast agents (Schmiedl et al., MRI of blood-brain barrier permeability in astrocytic gliomas: application of smali and large molecular weight contrast media, Magn. Reson. Med. 22: 288, 1991). By exploiting this violation of vascular permeability, diseased tissue can be discerned in high contrast compared to healthy tissue.

Apart from the brain and spinal cord, however, such a permeability barrier does not apply to the above-mentioned contrast agents (Canty et al., First-pass entry of nonionic agent into the myocardial extravascular space. Effects on radiographic estimate of transit time and blood volume. Circulation 84 : 2071,

1991). This means that the enrichment of the contrast agent is no longer dependent on the permeability of the blood vessels, but only on the size of the extracellular space of the corresponding tissues. Delineation of the vessels in relation to the interstitial space is not possible when using these contrast agents.

Especially for vascular imaging, it would be of particular importance that the contrast agent diffuses exclusively into the vascular space. Such a blood-pool-agent should make it possible, with the help of nuclear spin tomography, to separate well-perfused tissue from poorly perfused tissue and thereby diagnose ischemia. Also, infarct-prone tissue should be delineated based on its anemia from other healthy or ischemic tissue when vascular contrast is used. This is particularly important when, for example, differentiating a heart attack from ischemia.

Until now, most patients suspected of having cardiovascular disease (these diseases are a common cause of death in Western industrialized countries) have to undergo a demanding diagnostic examination. In angiotherapy, x-ray diagnostics using iodine-containing contrast agents are currently used in »· · · * · · * » φφφ • *·· » » _ ··*»» • · · · ·. These examinations are associated with various disadvantages, for example with the risk of radiation exposure, as well as with inconveniences and burdens, which mainly arise from the fact that compared to NMR contrast agents, iodine-containing contrast agents must be used in a significantly higher concentration.

There is therefore a need for NMR-contrast agents that can mark the vascular space (blood-pool-agent),

These compounds should be characterized by good tolerability and high efficiency (high degree of signal intensity in MRI).

Approaches to solving at least part of this problem by using complexing agents that are bound to macromolecules or biomolecules have so far had only very limited success.

Thus, for example, the number of paramagnetic centers in the complexes described in European patent applications No. 0 088 695 and 0 150 844 is not sufficient for acceptable image formation.

When the number of required metal ions is increased by multiple introduction of complexing units into a macromolecular biomolecule, this is associated with an intolerable influence on the affinity and/or specificity of this biomolecule (J. Nucl., Med. 24, 1158 (1983)).

Macromolecules may be generally suitable as contrast agents for angiography. Albumin-GdDTPA (Radiology 1987;

162: 205) for example, however, shows 24 hours after intravenous4

9 9 9 » *9 « ø · 9 ·

9*9 I

9 · 9 9 its application in rats enrichment in liver tissue, which amounts to practically 30% of the dose. Furthermore, only 20% of the dose is eliminated in 24 hours.

The macromolecular polylysine-GdDTPA (European Patent Application 0 233 619) has also proved capable as a blood-pool-agent. This compound consists of a mixture of molecules of different sizes depending on the preparation conditions. In excretion experiments on rats, it could be shown that these macromolecules are excreted unchanged by glomerular filtration by the kidneys. Depending on the synthesis conditions, polylysine-GdDTPA can also contain molecules that are so large that they cannot pass through the capillaries of the kidneys during glomerular filtration and thus remain in the body.

Macromolecular contrast agents based on carbohydrates, for example dextran, have also been described (European patent application, publication number 0 326 226). The disadvantage of these compounds is that they contain only about 5% of the signal-enhancing paramagnetic cations.

The polymers described in European Patent Application No. 0 430 863 already represent a step on the way to blood-pool-agents, because they no longer show the characteristic heterogeneity for the above-considered polymers in relation to the size and molecular weight of the molecules. However, it still remains an open requirement with regard to future complete excludability, tolerability and/or efficacy.

The task therefore arises to prepare for use new diagnostic means, especially for the recognition and localization of vascular diseases, which would not have the stated shortcomings. This task is solved by the present invention.

flfl · «a • flfl* ·

V · fl · a « flfl * • flfl m «fl fl « fl V *· fl fi flfl

The essence of the invention

It has been found that complexes consisting of nitrogen-containing cascade polymers, provided with complexing ligands, at least 16 element ions with sequence number 20 to 29, 30, 42, 44 or 57 to 83, as well as cations of inorganic and/or organic bases, amino acids or amino acid amides and which optionally contain acylated amino groups are surprisingly excellently suited for NMR and X-ray diagnostics, without exhibiting the aforementioned disadvantages.

The subject of the present invention are cascade polymer complexes of general formula I

A-{X-[Y-(Z-/VK _v / _z ) _y ] _x } _and (I) in which

A denotes the nitrogen containing cascade core of basal multiplicity a,

X and Y denote, independently of each other, a direct link or a cascading reproductive unit of reproductive multiplicity x, or y,

Z and V independently denote the cascading reproductive unit of reproductive multiplicity z , respectively w ,

K denotes the remainder of the complex-forming lark, and denotes the number 2 to 12 a * t · · « · * * * a a • a ··« a a a • a«a · a • a « ♦ a a »« x, y, z and w denote independently each number 1 to 4, with the restriction that at least two reproductive units are different and that multiplicities apply to the product

16sa.x.y.z,wí:64.

As cascade cores A are suitable:

nitrogen atom,

2' \ Γ Ί / ^u

CH ₂ — (CHjIjt— N-CH ₂ -(CH ₂ )—t/ ₂ ^z Li Jp ul 1

u u'

N-CH ₂ -CH — N

9 ^H 2

CH, (CHX (CH _; ) n4' ^CH :-CH,-N /

at ² p.m

OLCH, —N —CH ₂ CH, —N —CHCH

N —CH ₂ CH ₂ — N —CH ₂ -CH ₂ —N —CH ₂ CH ₂

-N/

CH,CH, — N — CH-CH —N —CH,CH

9 *

• «

9 «9

,AT

R C(M-N 'U

AT,

LI >1' and N^N

N ^Z Ν'

2/ / ^u

N \ 2 u « 0

in which

m a n stands for numbers 1 up to 10 P indicates number 0 to 10 At ¹ means Q! or E , At ² means Q ² or E , E means group

(^2)0 CH ₂ —N

whereas 0 means number 1 to 6, ^Q1 means hydrogen atom or ^Q2 means direct link,

M denotes an alkylene chain with 1 to 10 carbon atoms, which is optionally interrupted by 1 to 3 oxygen atoms and/or is optionally substituted by 1 to 2 oxo groups and

R° denotes a branched or unbranched alkyl radical of up to 10 carbon atoms, a nitro group, an amino group, a carboxylic acid group or a

at /

Μ—N \ 2 u where the number Q corresponds to the basal multiplicity a .

The simplest case of a cascade nucleus is represented by a nitrogen atom, whose three bonds (basal multiplicity a =

3) they are occupied in the first inner layer (generation 1) by three reproductive units X, or Y (when X means a direct link), or Z (when X and Y mean a direct link); formulated differently: the three hydrogen atoms of the corresponding ammonia cascade starter A(H) _a = NH^ are substituted by three reproductive units X , or Y , or Z . The number Q contained in the cascade core gives the basal multiplicity a .

Breeding units X, Y, Z and V contain groups

-NQ Q , where Q is a hydrogen atom or Q and Q is a direct bond. In a reproductive unit (for example X), the contained number Q corresponds to the reproductive multiplicity of this unit (for example x in the case of X). The product of all multiples of a.x.y.z.w gives the number of bound complexing residues K in the cascade polymer. The polymers according to the present invention contain at least 16 and at most 64 K residues in the molecule, which can bind one to a maximum of three (in the case of divalent ions), preferably one ion, of the element ·· *· ·· 4 · · Λ « · · * ·· · « · « * * · · · 9 · · and * * 4 · 4 · · ·«·· * · 4 · · · • · MM Μ « * 4 « * * » of the above serial numbers.

The last generation, that is, on coreplex-forming residues

The K coupled reproduction unit V , is connected to K via

NH-groups (-NQ Q with Q meaning a hydrogen atom and

O ² = direct bond), while the preceding reproduction units can be connected to each other both via -NHQ groups (for example, by acylation reactions) and also via -NQ Q groups (for example, by alkylation reactions).

The cascade polymer complexes of the present invention have a maximum of 10 generations (that is, more than just one X, Y and Z reproductive unit may be present in the molecule), but preferably 2 to 4 generations, with at least two of the reproductive units in the molecule being different .

As advantageous cascade cores A can be mentioned

such as fall under above given general formulas when m means the numbers 1 up to 3 , especially number 1, n means the numbers 1 up to 3 , especially number 1, P O means the numbers 0 means the number 1 up to 3 , especially number 1, M denotes a group -ch ₂ - , -CO- or -CH ₂ CO- and R° denotes a group -ch ₂ NU ¹ U ² , -CH ₃ or -N0 _? . As for example another advantageous k state : cascade starters A(H) _& is possible

(in parentheses is the basal multiplicity ta a , given for the case of the following monosubstitution, or disubstitution, used for the construction of subsequent generations)

tris(aminoethyl)amine (a=6, or 3) tris(aminopropyl)amine (a=6, or 3) diethylenetriamine (a=5, or 3) triethylenetriamine (a=6 , or 4) tetraethylenepentamine (a=7, or 5) 1,3,5-tris(aminomethyl)benzene (a=6, or 3) trimesic acid triamide (a=6 , or 3) 1,4,7-triazacyclononane (a=3) 1,4,7,10-tetraazacyclododecane (a=4) 1,4,7,10,13-Pentaazacyclopentadecane (a=5) 1,4,8,11-tetraazacyclotetradecane (a=4) 1,4,7,10,13,lO-Hexaazacyclooctadecane (a=6) 1,4,7,10,13,16,19,22,25,28-deaazacyclo- triacontane (a=10) tetrakis(aminomethyl)methane (a=8, or 4) 1,1,1-tris(aminomethyl)ethane (a=6 , or 3) tris(aminopropyl)-nitromethane (a=6, or 3) 2,4,6-triamino-1,3,5-triazine (a=6, or 3) 1,3,5,7-adamantanetetra-amide carboxylic (a=8, or 4) Acid amide 1,3', 5,5'-diphenyl ether- tetracarboxylic (a=8, or 4) 1,2-bis[phenoxyethane]-acid amide -3',3'',5',5''-tetracarboxylic (a=8, or 4) 1,4,7,10,13,16,21,24-octaaza.bicyclo- [8,8,8]hexacosane (a=6)

It should be pointed out that the definition as cascade core A and thus the division of the cascade core and the first reproductive unit is purely formal and can thus be chosen independently of the actual synthetic structure of the desired cascade polymer complexes. Thus, for example, the tris(aminoethyl)amine used in example 4 can be considered as a cascade •« ·· ► · * I core A (see the general formula given first for A, where m=n=p=l , U ¹⁼ E so=la , but also as a nitrogen atom (= asad nucleus A), which has three reproduction units of the formula q' / as the first generation

—CH ₂ —-CH ₃ —N \ 2

Q (see definition E) .

The cascade reproduction units X, Y, Z and V are independently determined formulas

E ,

U—N

Ό where ΐΛ means q! or E ,

1?

u9 v means 0 or E , while

E denotes the group _(CH ₂ ) ₀ __CH ₂ — N, while o denotes the number 1 to 6,

2 Q ¹ denotes a hydrogen atom or Q and denotes a direct bond, 'Q denotes an alkylene chain with 1 to 20 carbon atoms, which is optionally interrupted by 1 to 10 oxygen atoms and/or one to two -N(CO) groups ₍ jR , one up to two phenylene and/or one to two phenylene·oxyl residues and/or is optionally substituted by one to two oxo groups, thiooxo groups, carny • · « » 4 « » · 4 · i

4«4* · ·

4 boxy groups, alkylcarboxy groups having 1 to 5 carbon atoms in alkyl, alkoxy groups having 1 to 5 carbon atoms, hydroxy groups or alkyl groups having 1 to 5 carbon atoms, wherein q is 0 or 1 and

R represents a hydrogen atom, a methyl group or an ethyl group, which is optionally substituted by 1 to 2 hydroxy groups or one carboxyl group, represents a hydrogen atom or a group

U-N denotes a methine group =CH, while at the same time IP denotes a direct bond or the M group and has the meaning and

group U , or denotes a group

when simultaneously and are identical and denote a direct bond or group M .

Advantageous X, Y, Z and V cascade reproduction units

• · · · « · are those in which the U ³ residue in the above-mentioned general formulas denotes the group -CO-, -COCH ₂ OCH ₂ CO-, -COCH ₂ -, -CH ₂ CH ₂ -, -conhc ₆ h ₄ -, -coch ₂ ch ₂ co-, -coch ₂ -ch ₂ ch ₂ co-, -coch ₂ ch ₂ ch ₂ ch ₂ co-, residue direct bond or group -CH ₂ CO- , residue U ³ direct bond or groups -(CH ₂ ) ₄ -, -CH ₂ CO-, -CH(COOH)-, CH ₂ OCH ₂ CH ₂ -, -CH ₂ C ₆ H ₄ -, -CH ₂ -C ₆ H ₄ OCH ₂ CH ₂ - and the remainder E group q ¹ /

—CH ₂ —CH ₂ —N \ 2 Q

As examples of the mentioned cascade production units X, Y, Z and V, it is possible to mention:

-CH ₂ CH ₂ NH-; -CH ₂ CH ₂ N<; -COCH(NH-)(CH ₂ ) ₄ NH-;

-COCH(N<)(CH ₂ ) ₄ N<; -COCH ₂ OCH ₂ CON(CH ₂ CH ₂ NH-) ₂ ;

-COCH ₂ OCH ₂ CON(CH ₂ CH ₂ N<) ₂ ; -COCH ₂ N(CH ₂ CH ₂ NH-) ₂ ;

-COCH ₂ N(CH ₂ CH ₂ N<) ₂ ; -COCH ₂ NH-; -COCH ₂ N<;

-COCH ₂ CH ₂ CON(CH ₂ CH ₂ NH-) ₂ ; -COCH ₂ CH ₂ CON(CH ₂ CH ₂ N<) ₂ ;

-COCH ₂ OCH ₂ CONH-C ₆ H ₄ -CH[CH ₂ CON(CH ₂ CH ₂ NH-) ₂ 1 ₂ ;

-COCH ₂ OCH ₂ CONH-C ₆ H ₄ -CH[CH ₂ CON(CH ₂ CH ₂ N<) ₂ ] ₂ ;

-COCH ₂ CH ₂ CO-NH-C ₆ H ₄ -CH[CH ₂ CON(CH ₂ CH ₂ NH-) ₂ ] ₂ ;

-COCH ₂ CH ₂ CO-NH-C ₆ H ₄ -CH[CH ₂ CON(CH ₂ CH ₂ N<) ₂ ] ₂ ;

-CONH-C ₆ H ₄ -CH[CH ₂ CON(CH ₂ CH ₂ NH-) ₂ ] ₂ ;

-CONH-C ₆ H ₄ -CH[CH ₂ CON(CH ₂ CH ₂ N<) ₂ ] ₂ ; -COCH(NH-)CH(COOH)NH-; -COCH(N<)CH(COOH)N<;

• • · • • · in « • and • · « lips • lips lips • • · « • • lips • · • • lips • rtrt • « lips • · lips • · rtrt « • · • • rtrt lips • · • *·

,CON(CH ₂ CH ₂ NH-)

-COCH ₂ OCH ₂ CONH \\

CON(CH ₂ CH ₂ NH-) ₂

CON(CH ₂ CH ₂ N<) ₂ <och ₂ och ₂ conh

CON(CH ₂ CH ₂ N<) ₂

CON(CH ₂ CH ₂ NH-) ₂

CON(CH ₂ CH ₂ NH-) ₂ ,CON(CH ₂ CH ₂ N<) ₂

OONH'

CON(CH ₂ CH,N<) ₂ • »

CON(CH ₂ CH ₂ NH-) ₂

-coch ₂ ch ₂ conh —\

CON(CH ₂ CH ₂ NH-) ₂

CON(CH ₂ CH ₂ N<) ₂

-coch ₂ ch ₂ conh

_/ och ₂ ch ₂ n<

OCH, CH ₂ NHV /—OCH ₂ CH ₂ NH

OCH ₃ CH ₂ NH-

0 0 0 • 0 4 • · * 0 ·· «« ·· • · · · ♦ * • · · « • *000 • · 0 0 · *000 00 _e

O(CH ₂ CH ₂ O) ₂ CH ₂ CH ₂ NH—OíCKCH ₂ O) ₂ CH ₂ CH ₂ NH/ '' \

C(CH ₂ CH ₂ O) ₂ CK ₂ CH,NH-

Q(CH ₂ CH ₂ O) ₂ CH ₂ CH ₂ N<

O(CH ₂ CH ₂ O) ₂ CH ₂ CH ₂ N<

O(CH ₂ CH ₂ C) ₂ CH ₂ CH ₂ N<

The complexing residues K are described by the general formulas Ia and lb:

R ¹ OOC-R ² HC

N— CH ₂ — CH ₂ - - N

I ί ch ₂ ch ₂ ch ₂ ch,

I I

N—CH ₂ —CH ₂ —N

CHR-COOR ¹ ^CHR <0 0R _(lA y

R ⁵ i

ROCOH ₂ C CH-CO-a CH _? -COGR! there!

N—CH ₂ CH _; -n-CH ₂ CH ₂ -N ρ'ΟΟΟ-ΗΧ

CH ₂ -C OCR ¹ (IB)

- 19 • 9 9« ► · · 4 » 9 9«

9 9 « ι • 9 I • 9 9 in which independently of each other a hydrogen atom or the equivalent of a metal ion of serial number 20 to 29, 39, 42 to 44 or 57 to 83 , denotes a hydrogen atom, a methyl or an ethyl group, which is substituted with 1 to 2 hydroxy groups or one carboxy group,

R ^J denotes the group -CH-CO-NU-T, while υθ denotes a straight, branched, saturated or unsaturated alkylene group with 1 to 20 carbon atoms, optionally containing 1 to 5 imino groups, 1 to 3 phenylene groups, 1 to 3 phenyleneoxy groups, 1 to 3 phenyleneimino groups, 1 to 5 amido groups, 1 to 2 hydrazide groups, 1 to 5 carbonyl groups, 1 to 5 ethyleneoxy groups, one urea group, one thiourea group, 1 to 2 carboxyalkylimino groups, 1 to ester groups, 1 to 10 oxygen atoms , up to 5 sulfur atoms and/or 1 to 5 nitrogen atoms and/or optionally substituted with 1 to 5 hydroxy groups, 1 to 2 mercapto groups, 1 to oxo groups, 1 to 5 thioxo groups, 1 to 3 carboxy groups, 1 to 5 carboxyalkyl groups, 1 to 5 ester groups and/or 1 to amino groups, while optionally the contained phenylene groups can be substituted by 1 to carboxy groups, 1 to 2 sulfone groups • 9 ·* · · · • * 9 • · * • « #

9999 or 1 to 2 hydroxy groups,

T denotes the group -CO-α, -NHCO-a or -NHCS-a and and denotes the binding site on the terminal nitrogen atom of the last generation, reproductive unit

In .

Preferred complexing radicals K include those in which the above-mentioned formula IA contains an alkylene chain with 1 to 20 carbon atoms, preferably with 1 to 12 carbon atoms, in the sense of the U° group

-CH ₂ -, -CH ₂ NHCO-, -NHC0CH ₂ 0-, -NHCOCH ₂ OC ₆ H ₄ -, -N(CH ₂ C0 ₂ H)-, -nhcoch ₂ c ₆ h ₄ -, -nhcsnhc ₆ h ₄ -, -ch ₂ oc ₆ h ₄ -, ch ₂ -ch ₂ o-, and/or is substituted by the groups -COOH, -CH ₂ COOH.

The following groups can be cited as examples for:

-ch ₂ -, -ch ₂ ch ₂ -, -ch ₂ ch ₂ ch ₂ -, -c ₆ h ₄ -, -c ₆ h ₁₀ -, ch ₂ c ₆ h ₅ -, -CH ₂ NHCOCH ₂ CH( CH ₂ CO ₂ H)-c ₆ h ₄ -,

-ch ₂ nhcoch ₂ och ₂ -,

-ch ₂ nhcoch ₂ c ₆ h ₄ -,

-CH,NHCOCH ₉ G—C \ _ / every ₂ h

-ch ₂ nhcsnh-c ₆ h ₄ -CH(ch ₂ cooh)ch ₂ -, -CH ₂ 0C ₆ H ₄ -N(CH ₂ C00H)CH ₂ -,

-CH ₂ NHC0CH ₂ 0(CH ₂ CH ₂ 0) ₄ -C ₆ H ₄ -,

-ch ₂ oc ₆ h ₄ -,

Examples of the substituent include the following groups:

-ch ₃ , -c ₆ h ₅ , -ch ₂ -cooh,

-ch ₂ c ₆ h ₅ , -ch ₂ -o-(ch ₂ ch ₂ -o-) ₆ ch ₃ , -ch ₂ -oh.

When the composition according to the present invention is intended for use in NMR diagnostics, the central ion of the complex salt must be paramagnetic. These are especially the divalent and trivalent ions of the elements of sequence numbers 21 to 29, 42, and 58 to 70. Suitable ions are, for example, chromic, ferrous, cobalt, nickel, copper, praseodymium, neodymium, samarium and ytterbite. Due to their very strong magnetic moment, gadolinite, terbite, dysprosite, holmite, erbite, manganate and iron ions are particularly advantageous.

When the composition of the present invention is intended for use in X-ray diagnostics, the central ion must be derived from elements of higher order numbers in order to achieve sufficient X-ray absorption. Rylo found that for this purpose, diagnostic means are suitable, which contain a physiologically acceptable complex solution with a central ion from the elements of serial numbers 21 to 29, 39,

42, 44 and 57 to 83; this is, for example, the lanthanum ion and the lanthanide series ions mentioned above.

The cascade polymer complexes according to the present invention contain at least 16 ions of the above sequence number.

Residual acidic hydrogen atoms, that is, those that have not been substituted by the central ion, can be completely or partially replaced by cations of inorganic and/or organic bases, amino acids or amino acid amides.

Suitable inorganic cations are, for example, lithium, potassium, calcium, magnesium and especially sodium ions. Suitable organic base cations include, but are not limited to, cations of primary, secondary or tertiary amines such as ethanolamine, diethanolamine, morpholine, glucamine,

Ν,N-dimethylglucamine and especially N-methylglucamine. Suitable amino acid cations are, for example, cations of lysine, arginine and ornithine, as well as amides of otherwise acidic or neutral amino acids.

The compounds according to the present invention, which have a molecular weight of 10,000 to 80,000 D , preferably 15,000 to 40,000 D , have the already mentioned desired properties. They contain the large number of metal ions required for their use, stably bound in a complex,

They are enriched in areas with increased permeability of blood vessels, for example in tumors, they allow suction through the tissue pericytes, give the possibility to determine the volume of blood in the tissues, selectively shorten relaxation times, or blood density, and allow the permeability of the bloodstream to be visualized. Such physiological information cannot be obtained using extracellular contrast agents such as Gd-DTPA [Magnevist^] . From these points of view, the areas of use in modern image-providing procedures of nuclear spin tomography and computed tomography also result: specific diagnosis of malignant tumors, early control of therapy during cytostatic, antiphlogistic or vasodilatory therapy, early recognition of minimally perfused areas (for example in the myocardium), angiography in vascular diseases and recognition and diagnosis of (sterile or infectious) inflammations.

The cascade polymer complexes according to the present invention are also excellently suited for (interstitial and intravenous) lymphography.

As additional advantages compared to extracellular contrast agents such as Gd-DTPA [Magnevist^], it should be highlighted the higher efficiency as a contrast agent for nuclear spin tomography (higher relaxivity), which leads to a substantial reduction of the diagnostically necessary dose. At the same time, the contrast agents according to the present invention can be formulated as solutions iso-osmolar to the blood and thus reduce the osmotic load of the body, which is reflected in the reduced toxicity of the substance (higher toxicity threshold). Small doses and a higher toxicity threshold lead to a significant increase in the safety of the use of contrast agents in modern imaging methods.

Compared to macromolecular contrast agents based on carbohydrates, for example dextran (EP-A1-0 326 226), which, as already considered, usually carry only about 5% of the signal-enhancing paramagnetic cation, the polymer complexes according to the present invention have a content of • * * *0 * « · · · * * ··« · · · · * • · · · ♦ · * · ·«« · · saw about 20% of the paramagnetic cation. In this way, the macromolecules according to the present invention cause a significantly higher signal amplification for the molecule, which at the same time leads to the fact that the dose required for nuclear spin tomography is significantly lower compared to the dose of the macromolecule contrast agent based on carbohydrates.

Using the polymer complexes according to the present invention, it is possible to construct and produce macromolecules so that they have a uniformly defined molecular weight. It is surprisingly possible to control the size of the macromolecules so that they are large enough that they can leave the vasculature only slowly, but at the same time they are small enough to still pass through the capillaries of the kidneys, which are 300 to 800 A in size.

Compared to other considered polymer compounds according to the state of the art, the cascade polymer complexes according to the present invention are characterized by improved excretion ability, higher efficiency, greater stability and/or better acceptability.

Another advantage of the present invention is that complexes with hydrophilic or lipophilic, macrocyclic or open-chain, low molecular weight or high molecular weight ligands have now been made available. This makes it possible to control the tolerability and pharmacokinetics of these polymer complexes by chemical substitution.

The subject of the present invention is also a method for the production of cascade polymer complexes, the essence of which is that the compounds of the general formula I' « * * ·

A-{X-tY-(Z-/Vp _v / _z ) _y ] _x } _and (!') in which

A denotes the nitrogen containing cascade core of basal multiplicity a,

X and Y denote, independently of each other, a direct link or a cascading reproductive unit of reproductive multiplicity x, or y,

Z and V independently denote the cascading reproductive unit of reproductive multiplicity z , respectively w , and denote the number 2 to 12 , x, y, z and w independently denote the number 1 to 4 and β denotes the binding site of the terminal NH-groups of the last generation of the reproductive unit V , with the restriction that at least two reproduction units are different and that the multiplicities a hold for the product. x . y. of . w s 64 , with a complex or complexing substance K' of the general formula 1'A or 1'B » · ·

- 26 R ¹ OCC-R^HC » » · I « · · * · 0 I • · 4 « · » · 'N —CH, — CH, ~N

CH.

CH _Z

CH,

CH, (1'A)

N— CH ₂ — CH ₂ — N

CHR -CCOR

CHR -CCOR

ŘOOC-KC

^R5

CH-CHOCH ₂ -COOR'

N —CH ₂ CH ₂ —N—CH _? CH ₂ -N [ISk rooc-h,c ch ₂ -c oor while

Ί » _v

R denotes independently a hydrogen atom, the equivalent of a metal ion of sequence number 20 to 29, 39, 42 to 44 or 57 to 83, or an acid protecting group, o

R ^z represents a hydrogen atom, a methyl group or an ethyl group, which is optionally substituted by one or two hydroxy groups or one carboxy group • ·

- 27 ·· · « i * A · · A * · A · 9 A A · * * * A · · AAAA « • AAA AAA • AA· A « · « « · « a A

R ⁴ R ² denotes the group -CH-CO-NU^-T', while

R ⁴ denotes a straight or branched, saturated or unsaturated alkyl chain, which is optionally interrupted by 1 to 10 oxygen atoms, one phenylene or one phenylenoxy group and/or is optionally substituted by 1 to 5 hydroxy groups, 1 to 3 carboxy groups or one phenyl group, lj6 denotes a straight or branched, saturated or unsaturated alkylene group with 1 to 20 carbon atoms, optionally containing 1 to 5 imino groups, 1 to 3 phenylene groups, 1 to 3 phenoxy groups, 1 to 3 phenyleneimino groups, 1 to 5 amido groups, 1 to 2 hydrazide groups, 1 to 5 carbonyl groups, 1 to 5 ethyleneoxy groups, 1 urea group, 1 thiourea group, 1 to 2 carboxyalkylimino groups, 1 to 2 ester groups, 1 to 10 oxygen atoms, 1 to 5 sulfur atoms and/or 1 to 5 nitrogen atoms and /or optionally substituted with 1 to 5 hydroxy groups, 1 to 2 mercapto groups, 1 to 5 oxo groups, 1 to 5 thioxo groups, 1 to 3 carboxy groups, 1 to 5 carboxyalkyl groups, 1 to 5 ester groups and/or 1 to 3 amino groups, whereby the phenylene groups present may be substituted with 1 to 2 carboxy groups, 1 to 2 sulfone groups or 1

up to 2 hydroxy groups, a

Τ' denotes the group -C*0 , -COOH , -N=C=O or -N=C=S , denotes a hydrogen atom or a group and

C*0 denotes an activated carboxyl group, with the restriction that if K' denotes a complex, at least two (for divalent metals) or three (for trivalent metals) of the substituents R^ denote the equivalent of the metal ion of the above-mentioned elements and that they occur as needed, other carboxyl groups in the form of their salts with inorganic and/or organic bases, amino acids or amino acid amides, or the protective groups present are cleaved off, the cascade polymers obtained in this way - if K' denotes a complexing substance - are allowed to react in a known manner with at least with one metal oxide or metal salt of an element with sequence number 20 to 29, 39, 42, 44 or 57 to 83, and optionally then in the complexes of cascade polymers obtained in this way the acidic hydrogen atoms still present are completely or partially substituted by cations of inorganic and/or organic bases, amino acids or amides of amino acids, and optionally still present terminal amino groups are acylated as needed - before or after complexation with a metal.

Another object of the present invention are new compounds of general formula I'A

RCCC-fiHC

3'

R

N —CH, —CH ₂ —N ch ₂ ch ₂ ch ₂ ch ₂

N—CH ₂ —CH ₂ —N

O'A}

CHR ² COOR ¹ chr ² -coor ¹ in kterOm

R1 independently denotes a hydrogen atom, the equivalent of a metal ion of sequence number 20 to 29, 39, 42 to 44 or 57 to 83, or an acid protecting group,

R represents a hydrogen atom, a methyl group or an ethyl group, which is optionally substituted by one or two hydroxy groups or one carboxy group,

R ⁴ R ²

R ² denotes the group -CH-CO-NU^-T', whereby

R^ denotes a straight or branched, saturated or unsaturated alkyl chain, which is optionally interrupted by 1 to 10 oxygen atoms, one phenylene or one phenyleneoxy group and/or is optionally substituted by 1 to 5 hydroxy* » * · β · « fr ·» ·· «« ·> *1«

groups, 1 to 3 carboxy groups or one phenyl group, at ⁶ denotes a straight or branched, saturated or unsaturated alkylene group with 1 to 20 carbon atoms, optionally containing 1 to 5 imino groups, 1 to 3 phenylene groups, 1 to 3 phenoxy groups, 1 to 3 phenyleneimino groups, 1 to 5 amido groups, 1 to 2 hydrazide groups, 1 to 5 carbonyl groups, 1 to 5 ethyleneoxy groups, 1 urea group, 1 thiourea group, 1 to 2 carboxyalkylimino groups, 1 to 2 ester groups, 1 to 10 oxygen atoms, 1 to 5 sulfur atoms and/or 1 to 5 nitrogen atoms and/or optionally substituted with 1 to 5 hydroxy groups, 1 to 2 mercapto groups, 1 to 5 oxo groups, 1 to 5 thioxo groups, 1 to 3 carboxy groups, 1 to 5 carboxyalkyl groups, 1 to 5 ester groups and/or 1 to 3 amino groups. , whereby optionally present phenylene groups can be substituted by 1 to 2 carboxy groups, 1 to 2 sulfone groups or 1 up to 2 hydroxy groups, a T' denotes the group -C*0, -COOH, -N=C=O or -N=C=S , n5 R stands for a hydrogen atom or a group C*0 means activated carboxyl group.

These compounds serve as important intermediates for the production of cascade polymer complexes of general formula I.

• « ► flfl

F · « • fl · fl 4 fl · I

Anhydride, p-nitrophenyl ester, hydroxysuccinic acid imidester, pentafluorophenyl ester and acid chloride can be mentioned as examples of activated carbonyl groups C'0 in complexes, or complexing substances K'.

Additions or acylations, carried out to introduce complexing units, are carried out with substrates that contain the necessary substituent K (possibly bound to a cleavable group), or the required substituent is generated from them by reaction.

Examples of addition reactions include reactions of isocyanates and isothiocyanates, whereby the reaction of isocyanates is preferably carried out in an aprotic solvent, such as tetrahydrofuran, dioxane, dimethylformamide, dimethylsulfoxide and methylene chloride, at a temperature in the range of 0°C to 100°C, preferably 0° C to 50 °C, optionally with the addition of an organic base, such as triethylami η, pyridine, lutidine, N-ethyldiisopropylamine and N-methylmorpholine. The reaction with isothiocyanates is usually carried out in solvents such as water or lower alcohols such as methyl alcohol, ethyl alcohol, isopropyl alcohol! or mixtures thereof, dimethylformamide or mixtures of dimethylformamide and water, at a temperature in the range of 0 °C to 100 °C, preferably 0 °C to 50 °C, optionally with the addition of an organic or inorganic base, such as triethylamine, pyridine, lutidine, N -ethyldiisopropylamine, N-methylmorpholine or alkali or alkaline earth metal hydroxides, such as lithium hydroxide, sodium hydroxide, potassium hydroxide or calcium hydroxide, or carbonates of said metals such as magnesium carbonate.

Examples of acylation reactions include the reaction of free carboxylic acids using methods known to those skilled in the art (for example, J.P. Greenstein, M. Vinitz, Chemistry of the Amino Acids, John Viley & Sons, N, Y. (1961), p.

943-945). However, it has been shown to be advantageous to convert the carboxylic acid groups into an activated form prior to the acylation reaction, such as an anhydride, an active ester, or an acid chloride (eg, E. Gross, J. Meienhofer, The Peptides, Academie Press, N, Y. (1979), Vol. 1, pp. 65-314 N.F. React. 12, 157 (1962).

In the case of a reaction with an active ester, reference can be made to the literature known to those skilled in the art (for example, Houben Veyl, Methoden der organischen Chemie, Georg Thieme Verlag, Stuttgart, vol. E 5 (1985), 633). It can be carried out under the conditions mentioned above for the reaction with anhydrides. However, aprotic solvents such as methylene chloride and chloroform can also be used.

In the case of reactions with acid chlorides, only aprotic solvents are used, such as methylene chloride, chloroform, toluene or tetrahydrofuran at a temperature in the range of -20 °C to 50 °C, preferably 0 °C to 30 °C. known 1 iteration (for example Houben-Veyl, Methoden der Organischen Chemie, GeorgThieme-Verlag, Stuttgart (1974), vol. 15/2, pp. 355-364).

If R' represents an acid protecting group, lower alkyl, aryl and aralkyl groups are suitable, for example methyl group, ethyl group, propyl group, butyl group, phenyl group, benzyl group, diphenylmethyl group, triphenylmethyl group and also bis-(p-n i trophenyl.)-methyl group and trialkylsilyl groups.

• « * · V « · · • · ♦ ·

Optionally, the desired cleavage of protective groups is carried out by methods known to experts, for example hydrolysis, hydrogenolysis or alkaline saponification of esters with alkalis in a water-alcoholic solution at a temperature between 0 °C and 50 °C, or in the case of tert-butyl ester using trifluoroacetic acid.

Optionally, incompletely acylated terminal amino ligands or complexes can be converted to amides or semi-amides if desired. For example, it is possible to mention the reaction with acetic anhydride, succinic anhydride or diglycolic anhydride.

The introduction of the required metal ions is carried out, for example, in the manner described in DE-OS 34 01 052, by dissolving the oxide or salt (for example, nitrate, acetate, carbonate, chloride or sulphate) of the element with sequence number 20 to 29, 42, 44 or 57 to 83 in water and/or a lower alcohol (such as methyl alcohol, ethyl alcohol or isopropyl alcohol) and allowed to react with a solution or suspension of an equivalent amount of the complexing ligand and then, if desired, the acidic hydrogen atoms or acid groups present are substituted with inorganic cations and/or organic bases, amino acids or amino acid amides.

The introduction of the required metal joints can be carried out.

both at the level of the complexing substance IΆ or I'B , that is, before coupling to cascade polymers, and also after coupling of non-metalated ligands I'A or I'B .

bass,

In the case of, for example, hydroxides, neutralization is carried out using inorganic carbonates or hydrogen carbon- 34 • · • *

sodium, potassium, lithium, magnesium or calcium and/or organic bases such as, but not limited to, primary secondary and tertiary amines such as ethanolamine, morpholine, glucamine, N-methylglucamine and N,N-dimethylglucamine, as well as basic amino acids , such as lysine, arginine and ornithine, or amides of originally neutral or acidic amino acids, such as hippuric acid or glycine acetamide.

For the production of neutral complex compounds, for example, the desired base can be added to acid complex salts in an aqueous solution or suspension until the point of neutrality is reached. The resulting solution can then be concentrated to dryness under vacuum. It is often advantageous if the formed neutral salts are precipitated by the addition of water-miscible solvents, such as lower alcohols (methyl alcohol, ethyl alcohol, isopropyl alcohol, etc.), lower ketones (acetone, etc.) or polar ethers (tetrahydrofuran, dioxane, 1,2 -dimethoxyethane and the like), resulting in easy-to-isolate and easy-to-clean crystallites. Adding the desired bases to the reaction mixture already during the formation of the complexes has proven to be particularly advantageous, thus saving one reaction step.

When acid complex compounds contain more free acid groups, it is often expedient to produce neutral mixed salts that contain both inorganic and also organic cations as counterions.

This can be done, for example, by reacting complexing ligands in an aqueous suspension or solution with an oxide or salt of an element providing a central ion and half the amount of an organic base required for neutralization, the complex salt formed is isolated, purified as necessary, and then for complete neutralization, mix with the required amount of inorganic base. The order of bass additions can also be reversed.

The purification of the cascade polymer complexes obtained in this way is carried out after possibly adjusting the pH value by adding acid or base to 6 to 8, preferably 7, preferably by ultrafiltration with a membrane of suitable pore size (for example Amicon XM30, Amicon^ YM10, Amicon^ YM3) or by gel filtration at p

of suitable Sephadex gels.

In the case of neutral complex compounds, it is often advantageous to separate the ionic components through an anion exchanger, for example IRA 67 (OH”-form) and optionally additionally through a cation exchanger, for example IRC 50 (H ⁺ -form).

The production of terminal group-bearing cascade polymers, required for coupling to complexing substances K' (or also corresponding metals containing complexes), is generally based on commercially available, or can be produced according to literature data, nitrogen-containing cascade starters A(H) _and . The introduction of generation ions X, Y, Z and V is carried out using methods known from the literature (for example J. March, Advanced Organic Chemistry, 3rd ed.; John Viley & Sons, (1985), 364-381) by acylation or alkylation reactions with protected amines having the required structure, which contain functional groups capable of binding to the cascade nucleus, such as carboxylic acids, isocyanates, isothiocyanates or activated carboxylic acids (such as anhydrides, active esters, acid chlorides), or halides (such as chlorides, bromides , iodides) , aziridine, mesylate, tosylate or others, for • ·

cleavable groups known to those skilled in the art.

It has already been pointed out here that the distinction between the cascade core A and the reproductive units is purely formal. From a synthetic point of view, it may be advantageous not to use the formal cascade starter A(H) _and , but to introduce the nitrogen atoms, belonging by definition to cascade nuclei, only with the first generation. Thus, for example, for the synthesis of the compound described in example 1b), it is preferable not to alkylate the formal cascade nucleus of trimesic acid triamide with, for example, benzyloxycarbonylaziridine (six times), but to react trimesic acid trichloride with bis[2-(benzyloxycarbonylamino)-ethyl]-amine (three times) .

As protective groups of amino groups, it is possible to mention for experts the common benzyloxycarbonyl group, tert.-butoxycarbonyl group, trifluoroacetyl group, fluorenylmethoxycarbonyl group, benzyl group and formyl group (th. V. Greene, PGM Vuts, Protective Groups in Organic syntheses, 2nd Ed. John Viley & Sons (1991), pp. 309-385). After cleavage of these protecting groups, which is also carried out using methods known from the literature, the next desired generation can be introduced into the molecule. In addition to the construction of one generation consisting of two reaction steps (alkylation, or acylation and removal of protective groups), it is also possible to simultaneously introduce two, for example Χ-[Υ] _χ , or more generations, for example X-[Y- (Z) ] _χ . The construction of these multi-generational units is carried out by alkylation or acylation of unprotected amines with the structure of the desired reproductive units (reproductive amine) with a second reproductive amine whose amine groups are present in a protected form.

«0 ’ · · · • · · · · · • · * » ·

The compounds of general formula A(H) _and , required as cascade starter, are commercially available or can be prepared according to methods known from the literature or by analogy (for example, Houben-Veyl, Methoden der Org. Chemie, GeorgThieme-Verlag, Stuttgart (1957) , vol. 11/1; Inorg., 3702; TJ. Synth., 58 (1978); Heterocyclic Compounds: JS Bradshaw et al., Aza-CrownMacrocycles, John Víley & Sons, NY (1993)). Examples include:

Tris(aminoethyl)amine [e.g. Fluka Chemie AG, Schweiz; Aldrich-Chemie, Derutschland];

Tris(aminopropyl)amine [e.g. C. Voerner et al., Angew. Chem. Int. Ed Engl. (1993), 32, 1306];

Diethylenetriamine [e.g. fluke; Aldrich];

Triethylenetetramine [e.g. Fluke; Aldrich]; Tetraethylenepentamine [e.g. Fluke; Aldrich];

1,3,5-Tris(aminomethyl)benzene [e.g. TM, Farrett et al., J, Am. Chem. Soc. (1991), 113, 2965];

trimesic acid amide [e.g. H. Kurihara; Jpn. Kokai Tokkyo Koho JP 04077481; CA 117, 162453];

1,4,7-Triazacyclonan [e.g. Fluke; Aldrich];

1,4,7,10,13-Pentaazacyclopentadecane [e.g. K.V. Aston, Eur. Pat Appl. 0 524 161, CA 120, 44580];

1.4.7.10- tetraazacyclododecane (e.g. Aldrich);

1.4.8.11- Tetraazacyclotetradecane [e.g. Fluke; Aldrich]; 1,4,7,10,13,10,19,22,25,28 Decaazacyk.lot r i acontane (eg, A. Andres et al., J. Chem. Soc Dalton Trans. (1993), 3507]);

1,1,1-Tris(aminomethyl)ethane [e.g. R. J. Geue et al., Aus. J. Chem. (1983), 36, 927];

Tris(aminopropyl)-nitromethane [e.g. G.R, Newkome et al., φ« φφφφ φφφ φ

φ φ angew. Chem. 103. 1205 (1991) in analogy with RC Larock, Comprehensive Organic Transformations, VCH Publishers, N,Y, (1989), 419-420] 1,3,5,7-adamantanetetracarboxylic acid amide [e.g. Stetter et al., Tetr. Lett. 1967. 1841]; 1,2-B is [Phenoxyethane]-3 ¹ , 3 '' , 5 ' , 5 '' -tetracarboxylic acid amide [e.g. JP Collman et al.; J. Am. Chem. Soc. (1988), 110. 3477-86 by analogy from the prescription for example lb)];

1,4,7,10,13,16,21,24-Octaazabicyclo[8.8.8. ]hexacosane [e.g.

P.H. Smith et al., J. Org. Chem. (1993), 58 7939],

The production of reproductive amines, containing the above-mentioned functional groups, required for the construction of generations, is carried out according to the regulations described in the experimental part, or according to the methods described in the literature, or analogously.

For example, it is possible to state:

j^alpha, f4 ^e P ^s il° ⁿ -Di - CO-O-CH^C^Hg-Lysin-p-ni.tr of eny lester [recipe for example lc)];

hooc-ch ₂ och ₂ co-n(ch ₂ ch ₂ nh-co-o-ch ₂ c ₆ h ₅ ) ₂ ;

HOOC-CH ₂ N(CH ₂ CH ₂ NH-CO-O-CH ₂ C ₆ H ₅ ) ₂ ;

HOOC-CH ₂ CH ₂ CO-N(CH ₂ CH ₂ NH-COCF ₃ ) ₂ [produced according to the recipe for example 3a)] so that instead of bis(benzyloxycarbonylaminomethyl)amine it starts from bis (tr.if 1uoroacetylaminoethyl) amine and instead of diglycolic acid anhydride it is based on succinic anhydride] ;

»9 99 fa fa fa · · • fa · · ♦ fa fa fa fa fa fa 1 fa • V · fafafa fa·** • fa · «· · · fa fafafafa fa • fafa ·· fafafa *· fafafafa «· ·· · fafa fafa

HOOC-CH ₂ OCH ₂ CONH-C ₆ H ₄ -CH[CH ₂ CON(CH ₂ CH ₂ NH-CO-O-CH ₂ CgH ₅ ) ₂ ] ₂ [made according to the recipe for example 3a) ;

o=c=nc ₆ h ₄ -ch[ch ₂ con(ch ₂ ch ₂ nh-co-o-ch ₂ c ₆ h ₅ ) ₂ ] ₂

NH-CO-O-CH.CJ-I,

and. Q3

CON(CH ₂ CH ₂ NH-CO-O-CH.C.HJ,

HOOC-CH ₂ OCH ₂ CCNH

CON(CH ₂ CH ₂ NH-CO-O-CH ₂ C ₃ H

O=C=N

CON (CH ₂ CH ₂ NH-CO-O-CH ₂ C ₆ H ₅ ) ₂

CONíCH.CH„NH-CO-n-nn r M \ í _{£ 2} „g, , ₅ , ₂ «

« · • ·

N-benzyloxycarbonyl-aziridine prepared according to M. Zinice et al., J. Chem. Soc., Perkin Trans 1, 21-26 (1993);

N-benzyloxycarbonyl-glycine available for example from the company

Bach, California;

,oh ₂ ch ₂ nhco-o-ch ₂ c _s h ₅

HOOCOCKXH.NHCO-O-CH.C-FL

C (- tí o o

OCH ₂ CH ₂ NHCOO-CH ₂ C ₅ H ₅ made according to CJ Cavallita et al., J. Amer. Chem. Soc. 1943, 65, 2140, so that instead of benzyl chloride it is based on N-CO-O-Cl^C^H^-(2-bromomethyl)amine [AR Jacobson et al., J. Med. Chem. (1991),

34, 2816].

The production of complexes and complexing substances of the general formula ΙΆ and I'B is carried out according to the regulations described in the experimental part, or according to methods known from the literature (for example, EP 0 512 661 , 0 430 863 , 0 255 471 and 0 565 930 ) or analogously.

Thus, the production of compounds of the general formula J'A can proceed in such a way that the functional groups Τ'' serve as a precursor

• 9 group T'', either in the sense of a protected acid function, which can be converted independently of the acid protecting groups R into a free acid function using the above-mentioned methods, or in the sense of a protected amino function, which is deblocked by methods known from the literature [Th. V. Greene, P. G. M. Vuts, Protective Groups in Organic Synthesis, 2 nd edition, John Viley & Sons (1991), pp. 309-385] and can then be converted to isocyanates, or isothiocyanates [Methoden der Org. Chemistry (Houben Veyl),

E 4, pp. 742-749, 837-843, Georg Thieme Verlag, Stuttgart, New York (1983)]. Such compounds can be produced according to the regulations in the experimental part or analogously by monoalkylation of cyclenes using amides of α-halocarboxylic acids (in aprotic solvents such as chloroform).

The production of compounds of the general formula T'B can take place, for example, by using a protected acid function as a precursor to the activated carboxy group-C*O-, which is converted to a free acid function independently of the acid protecting group R according to the method described above and can also be activated according to methods described above, known from the literature. Such compounds can be produced according to the regulations given in the experimental part, possibly analogously, or, for example, by making an amino acid derivative of general formula II

H.N

C( ,v • 0 0 β

0 ·

0 0 · 0 *0 ·0 · 0 «

0 *

« 0 • 0* ·· *0 ♦ 0 0

0 0*0 0 0 »00 «0 •

0 in which

R ^J has the meaning given for R , whereby the hydroxy groups or carboxy groups contained in R are optionally present in a protected form and denote a straight or branched alkyl group with 1 to 6 carbon atoms, a benzyl group, a trimethylsilyl group, a triisopropylsilyl group,

2,2,2-trifluoroethoxy group or 2,2,2-trichloroethoxy group, wherein V is different from R is allowed to react with an alkylating agent of general formula III

CO ₂ R ^r ,N (NI)

Hal \^co _z r' in which ''

R stands for the protecting group and

Hal means a halogen atom, such as chlorine, bromine or iodine, preferably chlorine, [see M.A. Williams, H. Rapoport, J. Org. Chem. 58, 1151 (1993)].

Preferred amino acid derivatives are esters of naturally occurring α-amino acids.

• · • v

· · * · » · * · • · »4 * · · * · · ♦ · · · A • « A * * A ·

The reaction of the compound of the general formula II with the compound of the general formula III is preferably carried out as a buffered alkylation reaction, with an aqueous phosphate buffer solution serving as the buffer. The reaction is carried out at pH 7 to 9, but preferably 8. the buffer concentration can be 0.1 to 2.5 M, but a 2 M phosphate buffer solution is preferably used. The temperature during alkylation can be in the range of 0 °C to 50 °C, preferably room temperature.

The reaction is carried out in a polar solvent such as acetonitrile, tetrahydrofuran, 1,4-dioxane or 1,2-dimethoxyethane. Acetonitrile is preferably used.

The production of pharmaceutical compositions according to the present invention is also carried out using methods known per se, so that the complex compounds according to the present invention, optionally with the addition of the usual ingredients in the galenic, are suspended or dissolved in an aqueous medium and then the suspension or solution is sterilized. Suitable additives include, for example, physiologically harmless buffers (for example, tromethamine), additives of complexing substances or weak complexes (for example, diethyleneaminepentaacetic acid or corresponding complexes of Ca-cascade polymers), or if electrolytes are needed, for example sodium chloride, or if antioxidants are needed, for example ascorbic acid.

When suspensions or solutions of the compositions according to the present invention in water or physiological saline are required for enteral administration or other purposes, they are mixed with one or more excipients customary in galenics (for example, methylcellulose, lactose or • a · a «

R manniter) and/or surfactants (for example lecithin, Tween or Myrje^) and/or aromatic substances for taste correction (for example essential oils).

In principle, it is also possible to produce the pharmaceutical compositions according to the present invention also without isolating the complex salt. In any case, careful attention should be paid to the fact that the formation of the chelate is carried out in such a way that the salts according to the present invention and the salt solutions are practically free of non-complexed toxically acting metal ions.

This can be ensured, for example, by using color indicators such as xylenol orange, by control titration during the production process. The subject of the present invention is therefore also a method of producing complex compounds and their salts.

As a last resort, the purification of the isolated complex salt remains.

The pharmaceutical preparations according to the present invention preferably contain 1 gmol to 1.3 mol/1 complex salt and are usually dosed in an amount of 0.0001 to 5 mmol/kg.

They are intended for enteral and parenteral application. The compounds of the present invention are contemplated for use

1. for NMR diagnostics and X-ray diagnostics in the form of its complexes with the ions of elements with serial numbers 21 to 29, 39, 42, 44 and 57 to 83;

for radiodiagnosis and radiotherapy in the form of its complexes with radioisotopes of elements of sequence number 27,

29, 31, 32, 37 to 39, 43, 49, 62, 64, 70, 75 and 77.

- 45 • * · rtrt • * t · trt • * · ·♦ ** » «rt « • rt « · rt • · ♦ · · · · • · · 4 • ·» »· ··

The agents according to the present invention meet various prerequisites for suitability as contrast agents for nuclear spin tomography. Thus, they are perfectly suited to significantly improve the image obtained using nuclear spin tomography after oral or parenteral application by increasing the signal intensity. Furthermore, they exhibit the high efficiency required to burden the body with as small amounts of foreign substances as possible, as well as the good acceptability required to maintain the non-invasive nature of the tests.

The good solubility in water and the slight osmolality of the preparations according to the present invention allow the production of highly concentrated solutions and thereby maintaining the volumetric load of the circulation within acceptable limits and equalizing the dilution by body fluids. This means that NMR-diagnostics must be one hundred to one thousand times more soluble in water than for NMR-spectroscopy. Furthermore, the means according to the present invention show not only a high stability in vitro, but also a surprisingly high stability in vivo, so that the release or exchange of ions - poisonous in themselves - which are not covalently bound in the complexes, over time, in new contrast agents completely relearned, proceeds quite slowly.

In general, the compositions according to the present invention are dosed for use as NMR diagnostics in an amount of 0.0001 to 5 mmol/kg, preferably 0.005 to 0.5 mmol/kg. Details of use are discussed, for example, in H.J. Veinmann et al. . Am. J. of Roentgenology 142. 619 (1984).

Particularly low doses (below 1 mg/kg body weight) of organospecific NMR-diagnostics are useful, for example • # • 444 for the evidence of tumors and heart attacks.

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

The agents according to the present invention are suitable as radiodiagnostics based on their radioactive properties and the good stability of the complex compounds contained in them. Details about their use and dosage are described, for example, in the publication Radiotracers for Medical Applications, CRCPress, Boča Raton, Florida.

Another imaging method with radioisotopes is positron emission tomography, which uses positron emitting isotopes such as ^^Sc, ^^Sc, ^ ² Fe, and 68(3 _a (Heíss, VD,Phelps, Μ. E; Positron Em .ission Tomography of Brain, Springer Verlag Berlin, Heidelberg, New York, 1983) .

The compounds according to the present invention are surprisingly also suitable for the differentiation of malignant 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 are therefore well tolerated.

Since the substances according to the present invention are enriched in malignant tumors (no diffusion into healthy tissue, but high permeability through tumor vessels), they can also support the radiation therapy of malignant tumors. This differs from the corresponding diagnosis only in the amount and type of isotope used. The goal is to disrupt the cells * · » «V · I »· » « 9 9» * * · · · · « • · · * • · * « · *« of the tumor with energy-rich short-wave radiation with as little range as possible. For this, the interaction of metals contained in complexes (for example, iron or gadolinium) with ionizing radiation (for example, X-rays) or with neutron radiation is used. This effect significantly increases the local radiation dose at the place where the metal complex is located (for example, in a tumor). In order to achieve the same dose of radiation in malignant tissue, when using such metal complexes, the radiation load of healthy tissue can be substantially reduced, thereby reducing burdensome side effects for patients. Conjugates of metal complexes according to the present invention are therefore also suitable as radiosensitizing substances in the radiation therapy of malignant tumors (for example, by using the Mossbauer effect in neutron therapy). Suitable β-emitting ions are, for example, ⁴ ^Sc, ⁴ ^Sc, ⁴ ^Sc, ^ ² Ga, ^^Ga and . Suitable α-emitting

211 ions with small half-lives are, for example, Bi,

212bí, ²¹ ^Bí and ²¹⁴ Bi , with ²¹² Bi being preferred. A suitable photon and electron emitting ion is Gd, which can

157 obtained from Gd by neutron bombardment.

When the composition of the present invention is intended for use in a variant of the radiation therapy proposed by R.L. Mills et al. (Nature Vol. 336, (1988), p. 787) , so the central ion must be derived from a Mopbauer isotope such as ^^Fe or

When applying in vivo therapeutic agents according to " * J 1 * ' K τ F, , A 1 d >-» ρινυiOZCH CllkJ V J/ 1 ICA 1 C-Z. \ ) Cl O πΊ ΐ V Γ?v£ together* with a suitable carrier, such as, for example, serum or physiological sodium chloride solution, and together with another protein, such as, for example, human serum albumin. The dosage depends on the type of cellular disorder, on the metal ion used and on the type of imaging method.

The therapeutic agents according to the present invention are administered parenterally, preferably intravenously.

Details of the use of radiotherapeutics are discussed, for example, in the publication of R. V. Kozak et al., TIBTEC, Oktober 1986, 262 .

The compositions according to the present invention are excellently suitable as X-ray contrast agents, especially for computed tomography (CT), and it is particularly important to note that no anaphylaxis-like reaction symptoms, known from the use of iodine-containing contrast agents, can be detected with them in biochemical-pharmacological tests. . They are particularly valuable due to their good absorption properties in high voltage regions for digital subtraction techniques.

In general, the agents according to the present invention are dosed for use as X-ray contrast agents analogously to, for example, meglumine diatrizoate in an amount of 0.1 to 5 mmol/kg, preferably 0.25 to 1 mmol/kg.

Details of the use of X-ray contrast agents are discussed, for example, in Barke, Rontgenkontrastmitte.l , G. Thieme, Leipzig (1970) and P. Thurn, E. Bácneler Einfiihrung in die Rontgendiagnostik, G. Thieme, Stuttgart, New York (1977).

On the whole, it was possible to synthesize new complexing substances, metal complexes and metal complex salts, which open up new possibilities in diagnostic and therapeutic medicine49 no.

The following examples serve to clarify the object of the invention in more detail.

Examples of embodiments of the invention

Example 1

a) Bis[2-(benzyloxycarbonylamino)-ethyl]-amine

51.5 g (500 mmol) of diethylenetriamine and 139 ml (1 mol) of triethylamine are dissolved in dichloromethane and mixed with 161 g of benzyl cyanoformate (Fluka) in dichloromethane at -20 °C and the reaction mixture is stirred overnight at room temperature. After completion of the reaction, the mixture is evaporated, the obtained residue is taken up in diethyl ether, the organic phase is washed with sodium carbonate solution and dried using anhydrous sodium sulfate. The filtrate was mixed with hexane and the precipitate formed was filtered off and dried.

Yield: 163.4 g (88% of theory),

Elemental analysis calculated: found:

C 64.67 C 64.58

H 6.78 H 6.83

N 11.31

b) Ν,Ν,Ν',N',N'',N''-hexakis2-(benzyloxycarbonylamino)-ethyl]-trimesic acid triamide

13.27 g (50 mmol) of trimesic acid trichloride (Aldrich) and 34.7 ml (250 mmol) of triethylamine are dissolved in dimethylformamide and mixed at 0°C with 65.0 g (175 mmol) of the amine described in Example 1a) and the reaction mixture is then stirred overnight at room temperature. The obtained solution is evaporated in vacuo and the residue is chromatographed using ethyl acetate on silica gel.

Yield: 39.4 g (62% of theory).

Elemental analysis calculated: C 65.24 H 5.95 N 9.92 found: C 65.54 H 5.95 N 9.87

c) N ^a lf ^a ,N ^e P ^s il° ⁿ -bis(N,N'-dibenzyloxycarbonyl-lysyl)-lysine, protected tri-lysine

3.6 g (20 mmol) of lysine hydrochlorides and 6.95 ml (50 mmol) of triethylamine are dissolved in dimethylformamide, mixed with 26.8 g (50 mmol) ^{of Nalpha} , ^{Nepsilon -} dibenzyloxycarbonyl-lysine -p-nitrophenyl ester (Bachem) and the reaction mixture was stirred for 2 days at room temperature. After completion of the reaction, the mixture is evaporated in vacuo, the obtained residue is taken up in ethyl acetate and shaken with dilute hydrochloric acid. The organic phase is dried over anhydrous sodium sulfate, the solvent is evaporated, and the residue obtained is chromatographed using a mixture of ethyl acetate and ethyl alcohol with a graded gradient.

Yield: 10.7 g (57% of theory)

Elemental analysis calculated: light wall:

C 63.95 C 63.63

H 6.65 H 6.69

N8.95 N8.93

d) Fully protected benzyloxycarbonyl-24-polyamine based on Ν,Ν,Ν',N'',N''-hexakis[2-(trilysylamino)ethyl]-trimesic acid triamide

1.27 g (1 mmol) of hexa-benzylcarbonylamine, described in Example 1b), is dissolved in glacial acetic acid and mixed with 33% hydrogen bromide in glacial acetic acid with stirring. After 60 minutes, the starting precipitation is completed using diethyl ether, the resulting hexa-athnine hydrobromide is washed with diethyl ether, dried under vacuum and used without further purification in the reaction described below.

Yield: 0.95 g (quantitative).

7.0 g (7.5 mmol) of tri-lysine, described in example 1c), 1.2 g (7.5 mmol) of 1-hydroxybenzotriazole and 2.4 g (7.5 mmol) of 2-(1H-benzotriazole) -1-yl)-1,1,3,3-tetramethyluronium tetrafluoroborate (TBTU; Peboc Limited, UK) was dissolved in dimethylformamide and stirred for 15 minutes.

This solution was then mixed with 5.16 ml (30 mmol) of N-ethyldiisopropylamine and 0.95 g (1 mmol) of the hexaamine hydrobromide described above, and the reaction mixture was stirred overnight at room temperature. After completion of the reaction, the mixture is evaporated under vacuum and the obtained residue is chromatographed on silica gel using a mixture of ethyl acetate and ethyl alcohol (2:1).

Yield: 4.5 g (76% of theory).

Elementary analysis

N 10.52 N 10.29.

H 6.71 H 6.57

C 64.35 C 64.08 calculated: zj i s body:

e) Benzyl ester of 2-bromopropionylglycine

55.9 g (326.1 mmol) of chloride are added dropwise at 0 °C to 100 g (296.4 mmol) of glycine benzyl ester-p-toluenesulfonic acid salt and 33.0 g (326.1 mmol) of triethylamine in 400 ml of methylene chloride. of 2-bromopropionic acid, while not allowing the temperature to exceed 5 °C.

After the addition is complete, the reaction mixture is stirred for one hour at 0 °C and then for another 2 hours at room temperature. Next, 500 ml of ice water is added and the pH value of the aqueous phase is set to 2 using 10% aqueous hydrochloric acid. The organic phase is separated, washed once with 300 ml of 5% aqueous sodium carbonate solution and 400 ml of water, dried using anhydrous magnesium sulfate and evaporated to dryness under vacuum. The obtained residue is crystallized from diisopropyl ether.

Yield: 68.51 g (75% of theory) of colorless crystalline powder, melting point: 69 to 70 °C.

Elemental analysis calculated; found out:

C 46.76 C 46.91

H 7.19 N 4.54 Br 25.92 H 7.28 N 4.45 Br 25.81.

f) 1-[4-(benzyloxycarbonyl)-1-methyl-2-oxo-3-azabutyl]-1,4,/,10-LeLraazaoylk1ododcan

can, dissolved in 600 ml of chloroform, 50 g (162.2 mmol) of the compound mentioned in the title of example 1) is added and the reaction mixture is stirred overnight at room temperature. Then 500 ml of water is added, the organic phase is separated and

flfl·· e » · · • · · · is washed two more times with 400 ml of water. The organic phase is separated, dried over anhydrous magnesium sulfate and evaporated to dryness in vacuo. The obtained residue is chromatographed on silica gel using a mixture of chloroform/methyl alcohol/25% aqueous ammonia (10/5/1).

Yield: 40.0 g (63% of theory, based on compound 1e)) of a slightly yellowish viscous oil-like liquid.

Elemental analysis calculated: C 61.36 H 8.50 N 17.89 found: C 61.54 H 8.68 N 17.68.

g) 10-[4-(benzyloxycarbonyl)-1-methyl-2-oxo-3-azabutyl]-1,4,7-tris-(tert,-butoxycarbonylmethyl)-1,4,7,10-tetraazacyclododecane (sodium bromide complex)

To 20 g (51.08 mmol) of the compound mentioned in the title of example 1f) and 17.91 g (169 mmol) of sodium carbonate in 300 ml of acetonitrile is added 33 g (169 mmol) of bromoacetic acid tert-butyl ester and the reaction mixture is stir for 24 hours at a temperature of 60 °C. It is then cooled to 0 °C, the salts are filtered off and the filtrate is evaporated to dryness. The obtained residue is chromatographed on silica gel using a mixture of ethyl acetate/ethyl alcohol 15/1). The product containing fractions were evaporated and the residue obtained was

fl crystals suje from diisopropyietner. Yield : 34. .62 g (81% of theory) colorless crystalline • temperature thaw : 116 to 117 °C. powder

·· ·» •

Elemental analysis calculated: C 54.54 H 7.59 N 8.37 Na 2.74 Br 9.56 found: C 54.70 H 7.65 N 8.24 Na 2.60 Br 9.37

h) 10-(4-carboxy-1-methyl-2-oxo-3-azabutyl)-1,4,7-tris-(tert-butoxycarbonylmethyl)-1,4,7,10-tetraazacyclododecane (sodium bromide complex) g (35.85 mmol) of the compound mentioned in the title of example 1g) is dissolved in 500 ml of isopropyl alcohol and 3 g of palladium catalyst (10% Pd/C) are added. It is then hydrogenated overnight at room temperature, after which the catalyst is filtered off, the filtrate is evaporated to dryness in vacuo and the obtained residue is crystallized from acetone.

Yield: 22.75 g (85% of theory) of colorless crystalline powder, melting point: 225 °C (decomposition).

Elemental analysis calculated: C 49.86 H 7.69 N 9.38 Na 3.07 Br 10.71 found: C 49.75 H 7.81 N 9.25 Na 2.94 Br 10.58.

i) 24-mer N-(5-DO3A-yl-4-oxo-3-azahexanoyl)-cascade polyamide based on N,N,N',Ν',N'',N''-hexakis-12 acid triamide -(trilysylamino)-ethyl]-trimesine (where DO3A = 1,4,7-tris-(carboxymethyl)-1,4,7,10-tetraazacyclododecane)

6.0 g (1 mmol) of that in example 1d), and poly-benzyloxycarbonylamine, as described, is dissolved in glacial acetic acid with 33% hydrogen bromide in glacial acid.

vinegar. After 3 hours, the incipient precipitation is completed using diethyl ether, the resulting 24-amine hydrobromide is washed with diethyl ether and dried under vacuum.

35.84 g (48 mmol) of the acid described in the previous example 1h) are dissolved in dimethylformamide, mixed

7.35 g (48 mmol) of 1-hydroxybenzotriazole, 15.41 g (48 mmol) of TBTU (Peboc Limited, UK) and 49.3 ml (288 mmol) of N-ethyldiisopropylamine and the reaction mixture was stirred for 20 min at rooms. This solution was then mixed with the 24-amine hydrobromide described above (1 mmol) and stirred for 4 days at room temperature. The solution is then concentrated in vacuo, the remaining oil is cooled in an ice bath, mixed with trifluoroacetic acid, stirred overnight at room temperature and finally precipitated with diethyl ether. The precipitate is dried in a vacuum, taken out in water, the pH value is adjusted to 7, low molecular fractions are removed by filtration through an ultrafiltration membrane YM3 Amicon, and the retentate is finally membrane filtered and lyophilized.

Yield: 13.5 g (83% of theory) water content (Karl-Físcher): 6.2%

Elemental analysis (based on anhydrous substance) calculated: C 45.82 H 6.09 N 15.07 Na 10.79 found: C 45.56 H 6.15 N 14.80 Na 10.52 .

k) 24-mer-Gd-complex of N-(5-DO3A-yl-4-oxo-3-azahexanoyl)-cascade polyamide based on Ν,Ν,Ν' ,N',N'',N'' acid triamide - hexac i s-[2-(trilysyi amino)-ethyl]-trimesine

8.13 g (0.5 mmol) of complexing acid, described • ·

0 0 0 0 •0 0 00 • 0 • 0 0 0 • 0 * 0 0 • • · • • 0000 • 0 0 · • 0 0 • 0 0 0 0 0 0 0 0 « 0 0 0 0

in example li) , is dissolved in water, the pH value is adjusted to 3 with dilute hydrochloric acid and mixed

2.17 g (6 mmol) θ^2θ3 . The reaction mixture is stirred for 30 minutes at a temperature of 80 °C, after cooling, the pH value is adjusted to 7 and desalted on an ultrafiltration membrane YM3 AMICON lyof i 1 i suj e.

The retentate is finally membrane filtered and Yield: 8.89 g (92.1% of theory) water content (Karl-Fischer) : 9.6% Gd determination (AAS) : 19.6%

Elemental analysis (based on anhydrous calculated: C 40.26 H 5.35 N 13.24 found: C 39.98 H 5.51 N 13.42

Example 2

a) Benzyl ester of 2-bromopropionyl-p-alanine substance)

Gd 21.62 Gd 21.37 .

To 100 g (285 mmol) of β-alanine benzyl ester-p-toluenesulfonic acid salt and 31.67 g (313 mmol) of triethylamine in 400 ml of methylene chloride, 53.65 g (313 mmol) of 2-acid chloride are added dropwise at a temperature of 0 °C. bromopropion, while the temperature is not allowed to exceed 5 °C. After the addition is complete, the reaction mixture is stirred for one hour at 0 °C and then for another 2 hours at

1GUUVC VUUJ d JH/UUU VCA ptl aqueous hydrochloric acid separated, washed once with 300 ml of 5% aqueous hydrochloric acid, 300 ml of 5% aqueous sodium carbonate solution and 400 ml of water, dried using anhydrous magnesium sulfate and evaporated to dryness under vacuum . The obtained residue is crystallized from diisopropyl ether.

rooms. uaie se pxiua 500 mi the aqueous phase is adjusted with 10% dike to 2 . The organic phase se

AA > a A A • A · A

A A 1 • A AA

Yield: 71.36 g (78% of theory) of a colorless crystalline powder

Elemental analysis calculated: C 48.46 H 7.51 N 4.35 Br 24.80 found: C 48.29 H 7.65 N 4.25 Br 24.61.

b) 1-[5-(benzyloxycarbonyl)-1-methyl-2-oxo-3-azaheptyl]-1,4,7,10-tetraazacyclododecane

To 53.32 g (310 mmol) of 1,4,7,10-tetraazacyclododecane, dissolved in 600 ml of chloroform, 50 g (155.2 mmol.) of the compound mentioned in the title of example 2a) is added and the reaction mixture is stirred overnight at room temperature. Then 500 ml of water is added, the organic phase is separated and washed twice more with 400 ml of water. The organic phase is separated, dried over anhydrous magnesium sulfate and evaporated to dryness under vacuum. The obtained residue is chromatographed on silica gel using a mixture of chloroform/methyl alcohol/25% aqueous ammonia (10/5/1)

Yield: 38.39 g (61% of theory, based on compound 2a)) of a slightly yellowish viscous oil-like liquid.

Elemental analysis calculated: C 62.20 H 8.70 N 17.27 found: C62.05 H 8.81 N 17.15

c) 10-(5-(benzyloxycarbonyl)-1-methyl-2-oxo-3-azapentyl-1-1,4,7-tris(tert.-butoxycarbonyl-ethyl)-1,4,7,10tetraazacyl-ododecane (sodium bromide complex )

To 20 g (49.32 mmol) of the compound mentioned in the title of example 2b) and 17.28 g (163 mmol) of sodium carbonate in 300 ml of acetonitrile is added 31.8 g (163 mmol) of bromoacetic acid tert-butyl ester and reaction the mixture is stirred for 24 hours at a temperature of 60 °C. It is then cooled to 0 °C, the salts are filtered off and the filtrate is evaporated to dryness. The obtained residue is chromatographed on silica gel using a mixture of ethyl acetate/ethyl alcohol 10/1). The product containing fractions are evaporated and the residue obtained is crystallized from diisopropyl ether.

Yield: 31.89 g (76% of theory) colorless crystalline powder Elemental her analysis calculated : C 55.05 II 7.70 N 8.23 Na 2.69 Br 9.40 found out: C 55.17 H 7.85 N 8,10 Na 2.51 Bro 9.30

d) 10-[5-(carboxy)-1-methyl-2-oxo-3-azapentyl]-1,4,7-tris(tert-butoxycarbonylmethyl)-1,4,7,10tetraazacyclododecane (sodium bromide complex) g (35.26 mmol) of the compound mentioned in the title of example 2c) are dissolved in 500 ml of isopropyl alcohol and 3 g of palladium catalyst (10% Pd/C) are added. It is then hydrogenated overnight at room temperature, after which the catalyst is filtered off, the filtrate is evaporated to dryness under vacuum and the obtained residue is crystallized from acetone.

Yield: 24.41 g (91% of theory) of colorless crystalline powder • · · · 4 4 • · 4 4 * • · 4 · · 4 • · 4 * 4 • · 4 4 4 4 * · ·· · · 4 4 • · ♦ 44 • 4 444 4 4 • · 4 4 ··· 44 44

Elemental analysis calculated found:

C 50.52 H 7.82 N 9.21 C 50.41 H 7.95 N 9.10

At 3.01 Br 10.52 At 2.91 Br 10.37

e) 24-mer N-(5-D03A-yl-5-oxo-3-azaheptanoyl)-cascade polyamide based on Ν,Ν,Ν',N',N'',N''-hexakis-[ 2-(Trilysylamino)-ethyl]-trimesine %

Λ'

6.0 g (1 mmol) of poly-benzyloxycarbonylamine, described in example 1d), is dissolved in glacial acetic acid and mixed with 33% hydrogen bromide in glacial acetic acid with stirring. After 3 hours, the incipient precipitation is completed using diethyl ether, the resulting 24-amine hydrobromide is washed with diethyl ether and dried under vacuum.

36.52 g (48 mmol) of the acid, described in the previous example 2d.), are dissolved in dimethylformamide, mixed with

7.35 g (48 mmol) of 1-hydroxybenzotriazole, 15.41 g (48 mmol) of TBTU (Peboc Limited, UK) and 49.3 ml (288 mmol) of N-ethyldiisopropylamine and the reaction mixture was stirred for 20 minutes at rt rooms. This solution was then mixed with the 24-amine hydrobromide described above (1 mmol) and stirred for 4 days at room temperature. The solution is then concentrated in vacuo, the remaining oily substance is cooled in an ice bath, mixed with trifluoroacetic acid, stirred overnight at room temperature and finally precipitated with diethyl ether.

'•j, έ t The precipitate is dried in a vacuum, taken out in water, the pH value is '/z γ set to 7, by filtration through an ultrafiltration membrane YM3 í jj _e

Amicon gets rid of low molecular fractions and the retentate is finally membrane filtered and lyophilized.

Yield: 14.4 g (85% of theory) water content (Karl-Fischer): 8.7% ·· · · · ·« ·« • · · * ........

·· · · · · «ffffff • · · · · · · · *«« ff « • · · « · ffffff • · ♦ · · · · · ffffff ff | « «

Elemental analysis (based on anhydrous substance) calculated: C 46.82 H 5.98 N 14.79 Na 10.59 found: C 47.04 H 6.23 N 14.96 Na 10.26

f) 24-mer-Gd-complex of N-(6-DO3A-yl-5-oxo-4-azahexanoyl)-cascade polyamide based on acid triamide Ν,Ν,Ν',N',Ν'',N'' -hexakis-[2-(trilysylamino)-ethyl]-trimesine

8.5 g (0.5 mmol) of the complexing acid described in example 2e) are dissolved in water, the pH value is adjusted to 3 with dilute hydrochloric acid and 2.17 g (6 mmol) are mixed. The reaction mixture is stirred for minutes at a temperature of 80 °C, after cooling the pH value is adjusted to 7 and desalted on a YM3 AMICON ultrafiltration membrane. The retentate is finally membrane filtered and lyophilized.

Yield : 8.50 g (88% of theory) water content (Karl-Fischer) : 7.9% Gd determination (AAS) : 19.4%

Elemental analysis (based on anhydrous substance) calculated: C 41.12 H 5.52 N 12.99 Gd 21.21 found: C 40.86 H 5.35 N 13.25 Gd 20.95

Example 3

a) N,N'-bis(benzyloxycarbonyl)-3-[carboxymethoxyacety1]-3-azapentane-1,5-diamine

·· ···· • · · * · ♦ · 4 • 4 4«

37.14 g (100 mmol) of bis-(benzyloxycarbonyl-aminoethyl)-amine, described in Example 1a), are dissolved in dimethylformamide, mixed with 17.4 g (150 mmpl) of diglycolic acid anhydride (Jansen Chimica) in an ice bath and 21 mL (150 mmol) of triethylamine and the reaction mixture was stirred overnight at room temperature. The solution is then evaporated in vacuo, the residue obtained is taken up in ethyl acetate and shaken with dilute hydrochloric acid.

λ The organic phase is dried using anhydrous sodium sulfate and, after filtration, it is allowed to crystallize by adding hexane.

Yield: 41.4 g (85% of theory).

Elemental analysis calculated: C 59.13 H 6.00 N 8.62 found: C 58.99 H 5.93 N 8.70 .

b) N , N ' ,N ' ' ,N ' ' ' - tet.rakis{8 - (benzyl 1 oxy carbonylamino) - 6 - [ 2-(benzyloxycarbonylamino)-ethyl]-5-oxo-3-oxaocatnoyl } -cyclen

345 mg (2 mmol) of 1,4,7,10-tetraazacyclododecane (cyclen; Fluka) is azeotroped with toluene. A solution of 4.88 g (10 mmol) of N,N'-bis(benzyloxycarbonyl)-3-[carboxymethoxyacetyl]-3-azapentane-1,5-diamine ₄ (example 3a) is added to the cooled solution of cyclene in toluene at room temperature )) in tetrahydrofuran and 2.47 g (10 mmol)

of 2-ethoxy-1-ethoxycarbonyl-1,2-dihydroquinoline (EEDQ; Flu'ka) and the reaction mixture was stirred overnight. After the end of the reaction, the product is precipitated by the addition of hexane, the solvent is decanted off and the product is re-precipitated once more from a mixture of tetrahydrofuran/hexane and then tetrahydrofuran/toluene. After drying in a vacuum, 2.78 g (68% of theory) of yellowish ·· *· » * · « ·· 4» * 4 4 4 ·· ···· ·· · 4 1 • 4 <

«· · · solids.

Elemental analysis calculated: C 60.93 H 6.26 N 10.93 found: C 60.68 H 6.40 N 10.97

c) Fully protected benzyloxycarbonyl-32-polyamine based on 32-amine, condensed from

Ν,Ν',N'',N'''-tetrakis{8-(benzyloxycarbonylamino)-6-[2-(benzyloxycarbonylamino)-ethyl]-5-oxo-3-oxaokatnoyl}-cyclene and N ^a ^^ ^a ,N ^e P ^s ^^ ^on -b.is-(lysyl)-lysine (tri-lysine)

2.05 g (1 mmol) of octa-benzyloxycarbonylamine, described in example 3b), is dissolved in glacial acetic acid and mixed with 33% hydrogen bromide in glacial acetic acid with stirring. After 90 minutes, the incipient precipitation is completed with diethyl ether, the resulting octaamine hydrobromide is washed with diethyl ether, dried under vacuum and used without further purification in the reaction described below.

Yield: 1.6 g (quantitative)

9.4 g (10 mmol) of protected tri-lysine, described in example 1c), 1.5 g (10 mmol) of 1-hydroxybenzotriazole and 3.2 g (10 mmol) of 2-(1H-benzotriazol-1-yl) -1,1,3,3-tetramethyluronium tetrafluoroborate (TBTU; Peboc Limited, UK) was dissolved in dimethylformamide and stirred for 15 min. This solution was then mixed with 5.16 mL (30 mmol) of N-ethyldiisopropylamine and 1.6 g (1 mmol) of the above octamine hydrobromide and the reaction mixture is stirred overnight at rt. After completion of the reaction, the mixture is evaporated in vacuo and the obtained residue is chromatographed on silica gel for ·· ·· • · 9 fl

9 «I «999 « • 9 fl • 99 «9 ·♦ 99 ♦ · · 9 ι ΐ · · ♦ 9 9 9 « ♦ 9 9 fl

9999 99 using a mixture of dichloromethane/methyl alcohol (10:1), Yield: 6.0 g (72% of theory).

Elemental analysis calculated: C 63.32 H 6.26 N 10.93 found: C 60.68 H 6.40 N 10.97.

d) 32-mer N-(5-DO3A-yl-4-oxo-3-azahexanoyl)-cascade polyamide based on the 32-mer amine described in example 3c)

8.35 g (1 mmol) of 32-mer-benzyloxycarbonylamine, described in example 3c), is dissolved in glacial acetic acid and mixed with 33% hydrogen bromide in glacial acetic acid with stirring. After 3 hours, the incipient precipitation is completed using diethyl ether, the resulting 32-amine hydrobromide is washed with diethyl ether and dried under vacuum.

47.8 g (64 mmol) of the acid described in example 1h) are dissolved in dimethylformamide, mixed with 9.8 g (64 mmol) of 1-hydroxybenzotriazole, 20.5 g (64 mmol) of TBTU (Peboc Limited, UK ) and 65.7 mL (384 mmol) of N-ethyldiisopropylamine and the reaction mixture was stirred for 20 min at room temperature. This solution was then mixed with the above described (1 mmol) 32-amine hydrobromide and stirred for days at rooms. The solution is then concentrated in vacuo, the remaining oil is cooled in an ice bath, mixed with trifluoroacetic acid, stirred overnight at room temperature and finally precipitated with diethyl ether.

The precipitate is dried in a vacuum, taken up in water, the pH value is adjusted to 7, by filtration through an ultrafiltration membrane YM3 n

Amicon gets rid of low molecular fractions and the retentate is finally membrane filtered and lyophilized.

• 0

0 0 0 « * « « * * « 0 ♦' · 0

Yield: 17.2 g (76.4% of theory) water content (Karl-Fischer): 7.6%.

Elemental analysis (based on anhydrous substance calculated: C 45.73 H 6.12 N 15.08 Na 10.61 found: C 48.89 H 6.30 N 14.84 Na 10.31

e) 32-mer-Gd-complex N-(5-D03A-yl-4-oxo-3-azahexanoyl)-cascade polyamide based on the 32-mer amine described in example 3c)

10.4 g (0.5 mmol) of the complexing acid described in the previous example 3d) are dissolved in water, the pH is adjusted to 3 with dilute hydrochloric acid, 2.89 g (8 mmol) θ^2θ3 are mixed and stirred for 30 minutes at 80 °C. After cooling, the pH value is adjusted to 7 and desalted through an ultrafiltration membrane YM3 Amicon^ . The retentate is finally membrane filtered and lyophilized and pressed.

Yield: 12.1 g (91.1% of theory) water content (Karl-Fischer) : 11.0% Gd determination (AAS) : 18.6% .

Elemental analysis (based on anhydrous substance) calculated: C 40.26 H 5.39 N 13.28 Gd 21.30 found: C 40.10 H 5.21 N 13.04 Gd 21.03

An ytterbium complex is obtained in an analogous way with Y^ťCOj)^.

• · · · • * · · * · * 4 * * 4 * * · • * · •4 flfl··

Elemental analysis (based on anhydrous substance) calculated: C 39.42 H 5.28 N 13.00 Yb 22.94 found: C 39.29 H 5.40 N 12.81 Gd 22.65

Example 4

a) ester of hexaethylene glycol monomethyl ether-p-toluenesulfonic acid

To 20 g (67.49 mmol) of hexaethylene glycol monomethyl ether and 7.59 g (75 mmol) of triethylamine in 200 ml of chloroform, 14.3 g (75 mmol) of p-toluenesulfonic acid chloride are added in portions at a temperature of 0 °C, and the reaction mixture is stirred at this temperature for 4 hours. It is then evaporated to dryness under vacuum and the residue obtained is chromatographed on silica gel using a mixture of chloroform/methyl alcohol (5:1).

Yield: 27.67 g (91% of theory) of a scaly, glassy solid.

Elemental analysis calculated: C 53.32 H 7.61 S 7.12 found: C 53.15 H 7.70 S 7.03 .

b) 1-benzyloxy-5-(benzyloxycarbonyl)-2-chloro-3-oxo-4-azapentane

To 100 g (296.4 mmol) of the salt of glycine benzyl ester-p-toluenesulfonic acid and 33.0 g (326.1 mmol) of triethylamine in 400 ml of methylene chloride is added dropwise at a temperature • · «9 9**9

O °C 76 g (326.1 mmol) of 2-chloro-3-(benzyloxy)-propionic acid chloride (prepared according to Inorg. Chem., Vol. 31; 2422 (1992)) and stirred for 2 hours at this temperature. The reaction mixture is then mixed with 500 ml of ice water and the pH value is adjusted to 2 with 10% aqueous hydrochloric acid. The organic phase is separated, washed once with 300 ml of 5% aqueous hydrochloric acid, 300 ml of 5% aqueous sodium carbonate solution and 400 ml of water, dried using anhydrous magnesium sulfate and evaporated to dryness under vacuum. The obtained residue is chromatographed on silica gel using a mixture of methylene chloride/hexane/acetone (15:5:1). Yield ; 75.07 g (70% of theory)) of a slightly yellowish viscous oily substance.

Elemental analysis calculated: C 63.07 H 5.57 N 3.87 Cl 9.80 found: C 63.17 H 5.65 N 3.75 Cl 9.63

c) 1-(4-(benzyloxycarbonyl)-1-(benzyloxymethyl)-2-oxo-3-azabutyl]-1,4,7,10-tetraazacyclododecane g (193.5 mmol) of the compound mentioned in the title of example 4b) and 11.1 g (64.5 mmol) of 1,4,7,10-tetraazacyclododecane is dissolved in 70 ml of dimethylformamide and stirred for 2 days at a temperature of 50 °C. The reaction mixture is then evaporated to dryness, the obtained residue is taken up in 700 ml of water and extracted twice with 250 ml of chloroform each time. The organic phase is dried over anhydrous magnesium sulfate and evaporated to dryness in vacuo. The obtained residue is chromatographed on silica gel using a mixture of chloroform/methyl alcohol/25% aqueous ammonia (10:5:1).

Yield: 13.16 g (41% of theory, based on cyclene) viscous,

• · « · I fl fl flfl • « • « ♦ fl • fl • fl • fl « · • « fl • fl • • fl · fl » fl fl • • « « • » « « fl *flfl· • · fl · · «t • *

colorless oil-like substance.

Elemental analysis calculated: C 65.17 H 7.90 N 14.07 found: C 65.24 H 7.77 N 14.18

d) 10-[4-(benzyloxycarbonyl)-1-(benzyloxymethyl)-2-oxo-3-azabutyl]-1,4,7-tris(tert.-butoxycarbonylmethyl)-1,4,7,10-tetraazacyclododecane ( sodium bromide complex)

To 13 g (26.12 mmol) of the compound from example 4c) a

16.81 g (86.2 mmol) of tert-butyl bromoacetic acid is added to 9.14 g (86.2 mmol) of sodium carbonate in 200 ml of acetonitrile, and the reaction mixture is stirred for 24 hours at a temperature of 60 °C. It is then cooled to 0 °C, the salts are filtered off and the filtrate is evaporated to dryness. The obtained residue is chromatographed on silica gel using a mixture of ethyl acetate/ethyl alcohol (15:1).

Yield: 19.46 g (79% of theory) of a waxy solid.

Elemental analysis calculated: C 57.32 H 7.38 N 7.43 Na 2.43 Br 8.47 found: C 57.22 H 7.51 N 7.27 Na 2.33 Br 8.29

e) 10-[4-(carboxy-2-oxo-1-hydroxymethyl-3-azabutyl]-1,4,7-tris(tert-butoxycarbonylmethyl)-1,4,7,10-tetraaza-cyclododecane (sodium bromide complex)

To 19 g (20.15 mmol) of the compound from example 4d) in

3 g of palladium catalyst (10% Pd/C) are added to 300 ml of isopropyl alcohol and hydrogenated overnight at room temperature. *« ·0 0 * lote room. The catalyst is then filtered off, the filtrate is evaporated to dryness under vacuum and the obtained residue is crystallized from acetone.

Yield: 13.06 g (85% of theory) of a colorless crystalline powder.

Elemental analysis calculated: C 48.82 H 7.53 N 9.18 Na 3.00 Br 10.49 found: C 48.71 H 7.68 N 9.03 Na 2.81 Br 10.23

f) 10-[4-(benzyloxycarbonyl)-1-(hydroxymethyl)-2-oxo3-azabutyl]-1,4,7-tris(tert-butoxycarbonyl)-1,4,7,10-tetraazacyclododecane

To 13 g (17.04 mmol) of the compound from example 4e) a

3.42 g (20 mmol) of benzyl bromide is added to 6.11 g (18.75 mmol) of anhydrous cesium carbonate in 70 ml of dimethylformamide, and the reaction mixture is stirred overnight at a temperature of 50 °C. It is then cooled to 0 °C, 700 ml of water is added and extracted twice with 300 ml of methylene chloride each time. The organic phases are combined, washed twice with water, dried over anhydrous magnesium sulfate and evaporated to dryness in vacuo. The obtained residue is chromatographed on silica gel using a mixture of ethyl acetate/ethyl alcohol.

Yield: 9.97 g (78% of theory) of a colorless viscous oil.

Elemental analysis calculated: C 60.86 H 8.47 N 9.37 found: C 60.95 H 8.61 N 9.21 * * · fafa · fa ·«»· fa » · fa fafafa fafafa • * fafa· · fafa fafafa fafa fa·

g) 10-(4-(benzyloxycarbonyl)-1-(2,5,8,11,14,17,20-heptaoxa-heneicosanoyl)-2-oxo-3-azabutyl]-1,4,7-tris - (tert.-butoxycarbonylmethyl)-1,4,7,10-tetraazacyclododecane

9.7 g (12.93 mmol) of the compound from example 4f) is dissolved in 50 ml of tetrahydrofuran and 0.43 g (14.22 mmol) of sodium hydride (80% in paraffin) is added at a temperature of -10°C. The reaction mixture is stirred for 30 minutes at a temperature of 0 °C, after which 11.65 g (25.86 mmol) of the compound from example 4a) and 3.46 g (25.86 mmol) of lithium iodide are added. It is stirred for 24 hours at room temperature, after which 3 ml of water are carefully added and the mixture is evaporated to dryness. The obtained residue is chromatographed on silica gel using a mixture of chloroform/methyl alcohol (10:1).

Yield: 12.1 g (91% of theory) of a glassy solid.

Elemental analysis calculated: C 59.57 H 8.72 N 6.81 found: C 59.65 H 8.91 N 6.62

h) 10-(1-(2,8,11,14,17,20-heptaoxa-heneicosanoyl)-2-oxo-3-aza-4-(carboxy)-butyl]-1,4,7-tris- (tert.-butoxycarbonylmethyl)-1,4,7,10-tetraazacyclododecane g (11.67 mmol) of the compound from example 4g) is dissolved in 300 ml of isopropyl alcohol, 2 g of palladium catalyst (10% Pd/C) are added and hydrogenated overnight at room temperature. The catalyst is then filtered off, the filtrate is evaporated to dryness under vacuum and the obtained residue is crystallized from an acetone/diisopropyl ether mixture.

Yield: 10.18 g (93% of theory) of a waxy solid.

» 0 0 • 000 0 • ·

Elemental analysis calculated: C 56.33 H 8.92 N 7.46 found: C 56.20 H 9.03 N 7.35

i) 32-mer-Gd-complex N-(5-D03A-yl-4-oxo-3-aza-7,10,13,16,

19,22,25-heptaoxa-hexaoctanoyl)-cascade polyamide based on triamide of N,N,N',N',N'',N''-hexacid-[2-(trilysylamino)-ethyl]-trimesic acid

6.0 g (1 mmol) of the 24-mer-benzyloxycarbonylamine described in Example 1d) is dissolved in glacial acetic acid and mixed with 33% hydrogen bromide in glacial acetic acid with stirring. After 3 hours, the incipient precipitation is completed using diethyl ether, the resulting 24-amine hydrobromide is washed with diethyl ether and dried under vacuum.

45.03 g (48 mmol) of the acid described in example 4h) are dissolved in dimethylformamide, mixed with 7.35 g (48 mmol) of 1-hydroxybenzotriazole, 15.41 g (48 mmol) of TBTU (Peboc Limited , UK) and 49.3 mL (288 mmol) of N-ethyldiisopropylamine and the reaction mixture was stirred for 20 minutes at room temperature. This solution was then mixed with the 24-amine hydrobromide described above (1 mmol) and stirred for 4 days at room temperature. The solution is then concentrated in vacuo, the remaining oil is cooled in an ice bath, mixed with trifluoroacetic acid, stirred overnight at room temperature and finally precipitated with diethyl ether. The precipitate is dried under vacuum, taken up in water, the pH value is adjusted to 3 using dilute hydrochloric acid, mixed with 8.70 g (24 mmol), stirred for 4 hours at a temperature of 80 °C, after cooling the value The pH is adjusted to 7 by filtration through a YM3 Amicon ultrafiltration membrane

it gets rid of low molecular fractions and the retentate is finally membrane filtered and lyophilized.

Yield: 19.6 g (73.3% of theory) water content (Karl-Fischer) : 7.6% Gd determination (AAS) : 14.0%.

Elemental analysis (based on anhydrous substance) calculated found;

C 43.94 H 6.38 C 44.27 H 6.22

N 9.43 Cd 15.39 N 9.29 Gd 15.09

Example 5

a) 1,7-bis(trifluoroacetyl)-1,4,7-triazaheptane

113.3 g (790 mmol) of trifluoroacetic acid ethyl ester is added dropwise to a solution of 41.14 g (390 mmol) of 1,4,7-triazaheptane in 350 ml of tetrahydrofuran at a temperature of 80 °C under a nitrogen atmosphere, the reaction mixture is allowed to stir overnight at room temperature and then concentrated in vacuo. The oily liquid obtained is crystallized from hexane.

Yield: 115 g (99.9% of theory)

m.p. : 68 to 70 °C.

Elemental analysis calculated found;

C 32.55 H 3.76 C 32.63 H 3.75

F 38.62 N 14.24 F 38.38 N 14.19

b) 1,7 - bis (trifluoroacetyl) - 4- benzyloxy carbon 1. - 1,4,7- triazaheptane * · * * ·· · ·« • * * · »·« ·«·» • « » » «··· * * · · · * a « a · « » · • · · · · · · · I

M ···· > ··« · · l·

14.75 g (50 mmol) of the trifluoroacetyl compound, produced according to Example 5a) and 8.3 ml (60 mmol) of triethylamine are dissolved in 120 ml of dichloromethane and the solution is cooled to 0 °C. 7.5 ml (53 mmol) of benzyl ester of chloroformic acid are added with stirring. (97%) , dissolved in 20 ml of dichloromethane, the reaction mixture is allowed to stir overnight at room temperature, the salts are extracted using distilled water, the dichloromethane solution is dried using anhydrous sodium sulfate, it is concentrated to dryness in vacuo and the obtained residue is crystallized from mixture of diethyl ether and hexane.

Yield: 18.40 g (85.7% of theory)

m.p. : 131 to 132 °C.

Elemental analysis calculated: C 44.76 H 3.99 F 26.55 N 9.79 found: C 44.87 H 4.03 F 26.62 N 9.61

c) Di-tert-butyl ester of 3,9-bis(tert-butoxycarbonylmethyl)-6-benzyloxycarbonyl-3,6,9-triazaundecanedicarboxylic acid

4.29 g (10 mmol) of the trifluoroacetyl derivative prepared according to Example 5b) are dissolved in 30 ml of ethyl alcohol and mixed with 800 mg (20 mmol) of sodium hydroxide in 10 ml of distilled water. The reaction mixture is stirred for 3 hours at room temperature, concentrated to dryness at a bath temperature of 40 °C under vacuum, water residues are removed by azeotropic distillation with isopropyl alcohol and the obtained residue is taken up in 30 ml of dimethylformamide. 6.9 g (50 mmol) of potassium carbonate and 9.7 g (50 mmol) of bromoacetic acid tert-butyl ester are then added and 4-benzylcarbonyl-1,4,7-triazaheptane is alkylated overnight at room temperature. Dimethylformamide is then drawn off under the vacuum of an oil pump, the obtained residue is separated! between water and dichloromethane, the organic phase is separated, dried over anhydrous sodium sulfate, concentrated to dryness under vacuum and the residue obtained is purified by chromatography on silica gel. The title compound is eluted with a mixture of ethyl acetate and hexane. Yield: 6.49 g (93.6% of theory) of a foamy substance.

Elemental analysis calculated: C 62.32 H 8.57 N 6.06 found: C 62.41 H 8.66 N 6.01

d) Di-tert-butyl ester of 3,9-bis(tert-butoxycarbonylmethyl)-3,6,9-triazaundecanedicarboxylic acid

3.5 g (5 mmol) of the compound prepared according to example 5c) are dissolved in 100 ml of ethyl alcohol, 200 mg of Pearlman catalyst (20% Pd/C) are mixed and hydrogenated until the calculated amount of hydrogen is consumed. The catalyst is then aspirated and the liquid is concentrated to dryness under vacuum. The title compound is obtained in the form of a foamy substance. Yield: 2.80 g (99.9% of theory).

Elemental analysis calculated: C 60.08 H 9.54 N 7.51 found: C 60.02 H 9.62 N 7.56

e) Di-tert-butyl ester of 3,9-bis-(tert-butoxycarbonylmethyl)-6-[1-(ethoxycarbonyl)-ethyl-]-3,6,9-triazaunde-74

carboxyl candies

5.60 g (10 mmol) of the amino compound prepared according to example 5d) are dissolved in 30 ml of dimethylformamide, after which 1.66 g (12 mmol) of potassium carbonate and 2.17 g (12 mmol) of ethyl ester of acid 2 are added at room temperature -bromopropionate and stirred overnight. The reaction mixture is then poured into ice water, extracted with ethyl acetate, the organic phase is dried over anhydrous sodium sulfate and concentrated to dryness under vacuum. The title compound is obtained by chromatography on silica gel using a mixture of ethyl acetate and hexane.

Yield ; 4.18 g (63.4% of theory) .

Elemental analysis calculated: C 60.07 H 9.32 N 6.37 found: C 60.18 H 9.40 N 6.31

f) 3,9-bis(tert-butoxycarbonylmethyl)-6-[1-(carboxy)-ethyl-]-3,6,9-triazaundecanedicarboxylic acid di-tert-butyl ester

6.60 g (10 mmol) of the compound prepared according to example 5e) are dissolved in 50 ml of ethyl alcohol, after which a solution of 400 mg (10 mmol) of sodium hydroxide in 5 ml of distilled water is added and stirred for 3 hours at a temperature of 50 ° C. According to thin layer chromatography, saponification is quantitative. The mixture is then concentrated under vacuum, traces of water are removed by co-distillation with ethyl alcohol and the residue obtained is dried at 40°C under vacuum. The title compound is obtained as a white powdery substance. This residue is dissolved in 80 ml of moist ethyl alcohol (8:1) and for • · · · ·· * A » A · » · • » • A «

« A

I · · · • · « A 4

I « « • · · mixing is mixed with a solution of 535 mg (10 mmol) of ammonium chloride in 10 ml of distilled water. The reaction mixture is concentrated to dryness under vacuum, the soluble fractions are taken up in butyl alcohol and concentrated again to dryness under vacuum. The obtained residue is extracted with toluene. The organic solution is concentrated to dryness in vacuo and the title compound is obtained as a foam.

Yield: 5.35 g (84.7% of theory).

Elemental analysis calculated: C 58.93 H 9.09 N 6.65 found: C 59.01 H 9.16 N 6.60

g) 24-mer N-{N,N-bis[2-(N,N-bis-(carboxymethyl))-aminoethyl]-alanyl}-cascade polyamide based on sodium salt of Ν,Ν,Ν',Ν acid amide ',Ν'*,Ν''-hexacid[2-(trilysylamino)-ethyl]-trimesine

6.0 g (1 mmol) of the 24-mer-benzyloxycarbonylamine described in Example 1d) is dissolved in glacial acetic acid and mixed with 33% hydrogen bromide in glacial acetic acid with stirring. After 3 hours, the incipient precipitation is completed using diethyl ether, the resulting 24-amine hydrobromide is washed with diethyl ether and dried under vacuum.

30.33 g (48 mmol) of the acid described in Example 5f) was dissolved in dimethylformamide, 7.35 g (48 mmol) of 1-hydroxybenzotriazole, 15.41 g (48 mmol) of TBTU (Peboc Limited, UK) were mixed ) and 49.3 ml (288 mmol) of N-ethyldiisopropylamine and the reaction mixture is stirred for 20 minutes at This solution is then mixed with the above 24-amine hydrobromide and stirred for the time described at room temperature (1 mmol) • 9 • 9 » 9 9 4 » 9 9«

9 9 « 4 • 9 4 « 9 9 days at room temperature. The solution is then concentrated in vacuo, the remaining oily substance is cooled in an ice bath, mixed with trifluoroacetic acid, stirred overnight at room temperature and finally precipitated with diethyl ether. The precipitate is dried in a vacuum, taken out in water, the pH value is adjusted to 7, low molecular fractions are removed by filtration through an ultrafiltration membrane YM3 Amicon ^R , and the retentate is finally membrane filtered and lyophilized.

Yield: 11.0 g (86.3% of theory) water content (Karl-Fischer): 8.2%.

Elemental analysis calculated: C 42.87 H 5.41 N 11.96 Na 12.08 found: C 42.78 H 5.66 N 12.11 Na 11.89

h) 24-mer Gd complex of N-{N,N-bis-[2-(N,N-bis-(carboxymethyl))-aminoethyl]-alanyl}-cascade polyamide based on sodium salt of N,N,N acid triamide ',N',N'',N''-Hexakis[2-(tri1ysylamino)-ethyl]-trimesine

8.13 g (0.5 mmol) of the complexing acid described in the previous example 5g) are dissolved in water, the pH value is adjusted to 3 with dilute hydrochloric acid, 2.17 g (6 mmol) of Gd2C>3 are mixed and stirred for 30 minutes at 80 °C. After cooling, the pH value is adjusted to 7 and desalted through an ultrafiltration membrane YM3 p

Amicon. The retentate is finally membrane filtered and lyophilized.

Yield: 8.0 g (90.5% of theory) water content (Karl-Fischer); 7.5% Gd determination (AAS) : 21.0% .

• · · · « · ffff·· ff rt · · · * « · ···« · • ffff · · « ·« ·· ·*** *« ««· ·· ··

Elemental analysis calculated: C 35.93 H 4.38 N 10.03 Gd 23.09 Na 3.38 found: C 35.71 H 4.65 N 9.88 Gd 22.84 Na 3.50

Example 6

a) di-tert-butyl ester of 3,9-bis(tert-butoxycarbonylmethyl)-6-benzyloxycarbonylmethyl-3,6,9-triazaundecanecarboxylic acid

5.60 g (10 mmol) of the amino compound prepared according to the example are dissolved in 30 ml of dimethylformamide. 5d), after which 1.66 g (12 mmol) of potassium carbonate and 2.58 g (12 mmol) of bromoacetic acid benzyl ester are added at room temperature and the reaction mixture is stirred overnight. It is then poured into ice water, extracted with ethyl acetate, the organic phase is separated, dried over anhydrous sodium sulfate and evaporated to dryness under vacuum. After chromatography of the obtained residue on silica gel using a mixture of ethyl acetate and hexane, the title compound is obtained. Yield: 6.32 g (89.3% of theory).

Elemental analysis calculated: C 64.65 H 9.00 N 5.95 found; C 64.62 H 9.07 N 5.90

b) di-tert.-butyl ester of 3,9-b.is-(tert.-butoxy-carbonylmethyl)-6-carboxymethyl-3,6,9-1-riazaundecanedicarboxylic acid

7.08 g (10 mmol) of the compound prepared according to example 6a) are dissolved in 100 ml of ethyl alcohol, mixed with 0.4 g of Pearlman catalyst (20% Pd/C) and hydrogenated until 224 ml of hydrogen is consumed. The catalyst is then filtered off with suction, washed with ethyl alcohol and the liquid is concentrated to dryness under vacuum. The title compound is obtained as a foamy substance that crystallizes from a mixture of ethyl ether and hexane.

Yield: 6.87 g (97.3% of theory)

m.p. : 73 to 75 °C.

Elemental analysis calculated: C 57.85 H 9.00 N 5.95 found: C 57.91 H 9.11 N 6.01.

c) 32-mer N-{N,N-bis[2-(N,N-bis(carboxymethyl))-aminoethyl]-glycyl)-cascade polyamide based on the sodium salt of the 32-mer amine described in example 3c)

8.35 g (1 mmol) of 32-mer-benzyloxycarbonylamine, described in example 3c), is dissolved in glacial acetic acid and mixed with 33% hydrogen bromide in glacial acetic acid with stirring. After 3 hours, the incipient precipitation is completed using diethyl ether, the resulting 32-amine hydrobromide is washed with diethyl ether and dried under vacuum.

39.5 g (64 mmol) of the acid described in Example 6b) was dissolved in dimethylformamide, mixed with 9.8 g (64 mmol) of 1-hydroxybenzotriazole, 20.5 g (64 mmol) of TBTU (Peboe Limited, UK ) and 65.7 ml (384 mmol) of N-ethyldiisopropylamine and the reaction mixture was stirred for 20 minutes at room temperature. This solution is then mixed with the 32-amine hydrobromide described above (1 mmol) and stirred for days at room temperature. The solution is then concentrated in vacuo, the remaining oily substance is cooled in an ice bath, mixed with trifluoroacetic acid, stirred overnight at room temperature and finally precipitated with diethyl ether.

The precipitate is dried in a vacuum, taken up in water, the pH value is adjusted to 7, by filtration through an ultrafiltration membrane YM3 n

Amicon gets rid of low molecular fractions and the retentate is finally membrane filtered and lyophilized.

Yield: 15.7 g (78.6% of theory) water content (Karl-Fischer): 9.0%.

Elemental analysis calculated: C 41.77 H 5.24 N 12.33 Na 12.14 found: C 41.49 H 5.36 N 12.49 Na 11.93

c) 32-mer Gd-complex of N-{N,N-bis[2-(N,N-bis(carboxymethyl))-aminoethyl]-glycyl-cascade polyamide based on the sodium salt of the 32-mer amine described in example 3c )

10.0 g (0.5 mmol) of the complexing acid described in the previous example 6c) are dissolved in water, the pH value is adjusted to 3 with dilute hydrochloric acid, 2.89 g (8 mmol) of Gd2O are mixed and stirred for 30 minutes at a temperature of 80 °C. After cooling, the pH value is adjusted to 7 and desalted through a YM3 Amicon ultrafiltration membrane. The retentate is finally membrane filtered and lyophilized.

Yield: 10.9 g (90.9% of theory) water content (Karl-Fischer) : 9.5% Gd determination (AAS) 20.9%.

• · > · · ** ·«·t • · * ·

Elemental analysis (based on anhydrous substance) calculated: C 34.98 H 4.24 N 10.33 Gd 23.19 Na 3.39 found: C 35.20 H 4.08 N 10.46 Gd 22.89 Na 3 ,60

Example A

Comparison of in-vivo with extracellular contrast agent

The following experiment demonstrates the suitability of the compound described in example 1k) as a blood pool agent.

5 male rats (ScheringSPF) weighing 300 to 350 g are used as experimental animals. Before the experiment, the abdominal cavity is opened, the intestines are moved and then they are ligated through the posterior peritoneum with a surgical needle on both sides of the renal pelvis (arterial + venous). The abdominal cavity is then closed again. The animals are administered 0.3 ml (50 mmol/l) intravenously of a solution of the following contrast agent: a mixture of 1 part of the compound from example 1k), hereinafter referred to as compound 1, and the dysprosium complex 10-(1-hydroxymethyl-2,3-dihydroxypropyl )-1,4,7-tris(carboxymethyl)-1,4,7,10-tetraazacylododecane, produced according to EP 448 191, referred to below as compound 2. Through the catheter in the Arteria carotis communis, blood samples are taken at the following time points: 15,

30, 45, 60 and 90 sec., 3, 5, 10 and 15 minutes p.i.. In the obtained blood samples, the concentrations of gadol in.ia (Gd) and dysprosium (Dy) are measured in parallel using atomic emission spectrometry (ICP-AES). The proportion of injected contrast agent compound 1 (Gd) and compound remaining in the bloodstream

(Dy, comparator) can be compared by different labeling in the same animals. Since renal excretion is not possible, the decrease in blood concentration can only be attributed to dispersion in the bloodstream and diffusion into the interstitial tissue.

Results: the diffusion of compound 1 into the interstitium is significantly slowed down compared to the extracellular contrast agent of compound 2 (see Fig. 1).

Figure 1 shows the dependence of Gd (compound 1) and Dy (compound 2) blood concentration on time in rats (n=5) with ligated renal pelvises.

The extracellular contrast agent (compound 2) diffuses so rapidly into the interstitial space of the body that after only one minute p.i. equilibrium is reached (shown by a constant blood level). In contrast, with the cascade polymer (compound 1), not only is a higher concentration in the blood measured (evidence of a lower diffusion volume), but also the equilibria are not yet reached during the entire experimental period of 15 minutes (evidence of a very slow diffusion into the interstitial tissue). This means that compound 1 behaves as a blut-pool contrast agent.

Example B

Example of MR angiography in rabbits

The compound shown in example 1k) is tested on rabbits (CH. R. Kisslegg, =* 4 kg body weight) in an MR angiography experiment (Ganzkórper MRT System Siemens Vision, 1.5 Tesla, FISP 3D; TR : 400 ras ; flip angle : 45°;

·* ·» · ·· ··

coronal).

In the pre-contrast representation (see Fig. 2), only one or two larger vessels (for example, the subdominal aorta) are visible with relatively poor contrast (SI signal intensity of these vessels to the background) After intravenous administration of 50 pmol Gd/kg body weight of the compound described in for example lk) , a distinct increase in contrast (SI vessels/SI background) and a greater number of smaller and smallest vessels (for example a. and v. femoralis, a. and v. mesenterica caudalis, a. and v. renalis, a. and v. subrenalis and the like), which were not detectable before the administration of the contrast medium.

Example C

Example of lymph node enrichment in guinea pigs

The compound of the present invention, shown in example 1k), is tested 30 minutes to 24 hours after subcutaneous administration (10 pmol Gd/kg body weight, hind paw s.c.) on stimulated guinea pigs (complete Freund's adjuvant; each time 0.1 ml i.m. to right and left upper and lower thigh; 2 weeks before the administration of the test substance) taking into account its enrichment in the lymph nodes in three consecutive lymph nodes (popliteal, inguinal, iliac) In doing so, the results shown in the following table are obtained (findings concentration of Gd by ICP-AES) .

• ·

·· ·· • · · · 4 • 4 9 9 * • * * * · » • · · · « •4 4«»· 9«

Table

Time taken Ga-concentrations in three consecutive follow- in places of lymph nodes in pmol/1 (% dose/g tissue) mess up inguinal. 11 ikal. after 30 minutes p.i. 921 387 215 10:4,2:2,3 (20.1) (8.5) 4.7 90 minutes p.i. 659 120 68 10:1.8:1.0 (14.4) (2.6) (1.5) 4 o'clock p.i. 176 79 47 10:4,5:2,7 (3.9) (1.7) (1.0) 24 hours p.i. 62 13 28 10:2,1:4,5 (1.4) (0.3) (0.6)

• 9 9 9 • 9 a.m 99 • • 9 9 9 9 9 9 • • 9 « 9 9 ♦ 9 99 9 « • 9 « 9 9 9 9 9 9 9 9 9 9 9 9 9 • 99 • 9

• ·

9

1¾) cm diameter x gneiss

Claims (15)

PATENT CLAIMS 1. Complexes of cascade polymers, containing a) complexing ligands of general formula I A-{X-[Y-(Z-/VK _v / _z ) _y ] _x } _and (I), in which A denotes the nitrogen containing cascade core of basal multiplicity a, X and Y denote, independently of each other, a direct link or a cascading reproductive unit of reproductive multiplicity x, or y, Z and V independently denote the cascading reproductive unit of reproductive multiplicity z , respectively w , K is the residue of the complexing substance, and is the number 2 to 12, and x, y, z and w are independently the number 1 to 4, with the restriction that at least two reproductive units are different and that the product is multiplicities 16sa.x.y.z.wá64, b) at least 16 ions of elements with serial numbers 20 to 29, 39, • v · · • · · v • · · • · * * · t ·· ··»· » ^w · » : · · · > · · > ♦·· · · • · * »» ·· 42, 44 or 57 to 83, c) optionally cations of inorganic and/or organic bases, amino acids or amino acid amides, as well as d) optionally acylated xerminal amino groups. 2. Complexes of cascade polymers according to claim 1, characterized in that A denotes a nitrogen atom, ,/ N—CH ₂ —(CH ₂ )qN-CH ₂ -(CH ₂ )~ N LT N-CHj-CH —N CH, (CH CH., (CH,) N-^-CHj-CH ₂ -: / ^z '2 d p Lf 4 4 4 4 · 4 · · 4 4 4 » «· · 44 * « 4 4 4 « * 4 # · 4 · 4 4 4 4 4 4 • « * 4 · 4 4 « 4 •4 4444 44 4·4 44 «4 —CH ₂ -CH ₂ —Ν —CH ₂ CH ₂ Ν —CH ₂ CH ₂ Ν —CH ₂ -CH ₂ —Ν —CH ₂ CH ₂ -Ν ch ₂ ch ₂ —Ν — CH ₂ -CH ₂ —Ν —CH ₂ CH ₂ ^ζ ! ! I 2 I 2 υ υ R°C(M • 0 0 * * · 0 « · · · 0 • 0 0 *0 •0 0*00 00 4 · « i 0 0 0 0 • 4 4 »* 0· Φ » 1 ^u \ N U .u / \ in which man denotes the numbers 1 to 10, p w denotes the number 0 to 10, U ^J u ² denotes Q ¹ or E , denotes Q ² or E , denotes the group ,Q (CH ₂ ) _o —ch ₂ —n 4 « whereas O means number 1 to 6, About ¹ means hydrogen atom or ^Q2 means direct link, M denotes an alkylene chain with 1 to 10 carbon atoms, which is optionally interrupted by 1 to 3 oxygen atoms and/or is optionally substituted by 1 to 2 oxo groups and R° denotes a branched or unbranched alkyl radical with 1 to 10 carbon atoms, a nitro group, an amino group, a carboxylic acid group or an i group at / M-N \ 2 u V · V -r W 9 where the number Q corresponds to the basal multiplicity a . 3. Complexes of cascade polymers according to claim ί, characterized in that the cascade reproduction units X, Y, Z and V independently denote E. 0 0 0 ··· 0 U—N via Ijl stands for Q ¹ or E , 9 9 U stands for Q or E, whereas E denotes the group —<CH ₂ ) _o — CH,— N where 'Q t · * * · · · · » · · • * fl ^r * * fl fl··# • · · ··· · · « • · · · · · · · ··· · · · « o means Number 1 to 6, 1 <- 2 Q ¹ means a hydrogen atom or 0 and AND Q denotes a direct bond, ir denotes an alkylene chain with 1 to 20 carbon atoms, which is optionally interrupted by 1 to 10 oxygen atoms and/or one to two -N(CO)q-R , one to two phenylene and/or one to two phenylenoxy groups residues and/or is optionally substituted by one to two oxo groups, thiooxo groups, carboxy groups, alkylcarboxy groups with 1 to 5 carbon atoms in alkyl, alkoxy groups with 1 to 5 carbon atoms, hydroxy groups or alkyl groups with 1 to 5 carbon atoms, where q is 0 or 1 a R denotes a hydrogen atom, a methyl group or an ethyl group, which is optionally substituted by 1 to 2 hydroxy groups or one carboxyl group, L represents a hydrogen atom or a u group 3 / U-N \ 2 u V denotes the methine group =CH, when it simultaneously denotes a direct bond or the M group and has the meaning of the iP group, or • · » · · · V stands for group CO / - NH C0when at the same time and are identical and denote a direct bond or group M . 4. Complexes of cascade polymers according to claim 1, characterized in that the complex-forming residue K, bound to the terminal nitrogen atoms of the last generation of the reproduction unit V, is a residue of the general formula IA or IB R COOR HC N“CH ₂ —ch ₂ —N ch ₂ CH CH ₂ N—CH ₂ —CH ₂ CHR -COOR • » « fafa ROOC-H ₂ C FT CH-COch ₂ -coor N —CH ₃ CH ₂ -N-CH ₃ CH ₂ N ROOC-HjC CH ₂ -C OOR (IB) in which r! independently means a hydrogen atom or the equivalent of a metal ion of serial number 20 to 29, 39, 42 to 44 or 57 to 83, R denotes a hydrogen atom, a methyl or ethyl group, which is optionally substituted by 1 to 2 hydroxy groups or one carboxy group, R ⁴ R ² I I denotes the group -CH-CO-N-U^-T, while it denotes a straight, branched, saturated or unsaturated alkylene group with 1 to 20 carbon atoms, optionally containing 1 to 5 imino groups, 1 to 3 phenylene groups, 1 to 3 phenyleneoxy groups, 1 up to 3 phenyleneimino groups, 1 to 5 amido groups, 1 to 2 hydrazide groups. 1 to 5 carbonyl groups, 1 to 5 ethyleneoxy groups, one urea group, one thiourea group, 1 to 2 carboxyalkylimino groups, 1 to 2 ester groups, 1 to 10 oxygen atoms, 1 to 5 sulfur atoms and/or 1 to 5 nitrogen • 0 0 0 « «·* 000 « φ ·· · 0 · 00 000 atoms and/or optionally substituted by 1 to 5 hydroxy groups, 1 to 2 mercapto groups, 1 to 5 oxo groups . to 2 hydroxy groups, means a straight, branched, saturated or unsaturated alkyl chain with 1 to 30 carbon atoms, which is optionally interrupted by 1 to 10 oxygen atoms, 1 phenylene or 1 phenyleneoxy group and/or is optionally substituted by 1 to 5 hydroxy groups, 1 to 3 carboxy groups or 1 phenylene group, T denotes the group -CO-α , -NHCO-α or -NHCS-a and a denotes the binding site on the terminal nitrogen atom of the last generation, the reproduction unit V and R^ represents a hydrogen atom or a group R^ . 5. Cascade polymer complexes according to claim 4, characterized in that the alkylene chain with 1 to 20 carbon atoms contains groups -ch ₂ -, -ch ₂ nhco-, -nhcoch ₂ o-, -nhcoch ₂ oc ₆ h ₄ -, -N(CH ₂ CO ₂ H)-, -NHCOCH ₂ C ₆ H ₄ -, -nhcsnhc ₆ h ₄ -, -ch ₂ oc ₆ h ₄ -, ch ₂ -ch ₂ o-, a · ·* aa • · · a • · a · • · · · a « • · a aa ♦< and/or is substituted by -COOH, -CH2COOH groups. 6. Cascade polymer complexes according to claim 4, characterized in that U ⁶ denotes groups -ch ₂ -. -ch ₂ ch ₂ -, -ch ₂ ch ₂ ch ₂ -, -c ₆ h ₄ -, -c ₆ h ₁₀ -, ch ₂ c ₆ h ₅ -, -CH ₂ NHCOCH ₂ CH(CH ₂ CO ₂ H )-c ₆ h ₄ -, -ch ₂ nhcoch ₂ och ₂ -, -ch ₂ nhcoch ₂ c ₆ h ₄ - . 7. Complexes of cascade polymers according to claim 3, characterized in that the residue U contained in cascade reproduction units X, Y, Z and V represents the groups -CO-, -COCH ₂ 0CH ₂ CO-, -COCH ₂ -, - CH ₂ CH ₂ -, -CONHC ₆ H ₄ -, -COCH ₂ CH ₂ CO-, -COCH ₂ -CH ₂ CH ₂ CO- or -coch ₂ ch ₂ ch ₂ ch ₂ co-, the remainder is a direct bond or the group -CH ₂ CO-, the remainder is a direct bond or the group -(CH ₂ ) ₄ -, -CH ₂ CO-, -CH(COOH)-, CH ₂ OCH ₂ CH ₂ -, -CH ₂ C ₆ H ₄ -, -CH ₂ -C ₆ H ₄ OCH ₂ CH ₂ - and Q residue E group / —CH ₂ —CH ₂ ~N \ _Q ² i 2 where Q and Q have the above meaning, 8. Cascade polymer complexes according to claim 3, characterized in that the cascade reproductive units X, Y, Z and V independently represent groups -CH ₂ CH ₂ NH-; -CH ₂ CH ₂ N<; -COCH(NH-)(CH ₂ ) ₄ NH-; -COCH(N<)(CH ₂ ) ₄ N<; -COCH ₂ OCH ₂ CON(CH ₂ CH ₂ NH-) ₂ ; ·» · 4 4 4 ’ · «« • 4 4 *4 4 4 4 · 4 4 Β * •44 4 4 4 44« • 4 ··♦· 44 4 44 «β -COCH ₂ OCH ₂ CON(CH ₂ CH ₂ N<) ₂ ; -COCH ₂ N(CH ₂ CH ₂ NH-) ₂ ; -COCH ₂ N(CH ₂ CH ₂ N<) ₂ ; -COCH ₂ NH-; -COCH ₂ N<; -COCH ₂ CH ₂ CON(CH ₂ CH ₂ NH-) ₂ ; -COCH ₂ CH ₂ CON(CH ₂ CH ₂ N<) ₂ ; -COCH ₂ OCH ₂ CONH-C ₆ H ₄ -CH[CH ₂ CON(CH ₂ CH ₂ NH-) ₂ ] ₂ ; -COCH ₂ OCH ₂ CONH-C ₆ H ₄ -CH[CH ₂ CON(CH ₂ CH ₂ N<) ₂ ] ₂ ; -coch ₂ ch ₂ co-nh-c ₆ h ₄ -ch[ch ₂ con(ch ₂ ch ₂ nh-) ₂ ] ₂ ; -coch ₂ ch ₂ co-nh-c ₆ h ₄ -ch[ch ₂ con(ch ₂ ch ₂ n<) ₂ j ₂ ; -CONH-C _ó H ₄ -CH[CH ₂ CON(CH ₂ CH ₂ NH-) 2 ] 2 ; -CONH-C ₆ H ₄ -CH[CH ₂ CON(CH ₂ CH ₂ N<) ₂ ] ₂ ; -COCH(NH - ) CH (COOH)NH - ; -COCH(N<)CH(COOH)N<; -COCH, OCH, CONH -COCH ₂ OCH ₂ CONH _from CON(CH ₂ CH ₂ NH-) ₂ CON(CH ₂ CH ₂ NH-) ₂ CCN(CH ₂ CH ₂ N<). CON(CH ₂ CH ₂ N<) ₂ CCN(CH ₂ CH ₂ NH-) ₂ - 95a _/ CON(CH ₂ CH ₂ N<) ₂ - CONH W CON(CH ₂ CH ₂ N<) ₂ -coch ₂ ch ₂ conh CON(CH ₂ CH ₂ NH-)j CON(CH ₂ CH ₂ NH-) ₂ - COCH, CH _? CONH CON(CH ₂ CH ₂ N<) ₂ CON(CH ₂ CH ₂ N<) ₂ • Φ • · ·* • · »· * • · « · • · · • · • « Φ ♦ « · • » « · « « « • · • · • • · * · • * • · • • · · * » • · • t oh ₂ ch ₂ nhoch ₂ ch ₂ nh- O(CH ₂ CH ₂ O) ₂ CH ₂ CH ₂ NHO(CH ₂ CH ₂ O) ₂ CH ₂ CH ₂ NHO(CH ₂ CH ₃ O) ₂ CH ₂ CH ₂ NH/ // / λ O(Ch ₂ ch ₂ O) ₂ ch ₂ ch ₂ n< -C(CH ₂ CH ₂ O) ₂ CH ₂ CH ₂ N< O(CH ₂ CH ₂ O) ₂ CH ₂ CH ₂ N< 9. Complexes of cascade polymers according to claim 2, on what's up c í with E th m , that m means what's up 1 until 3 , n means numbers 1 until 3 , P means numbers 0 until 3 , O means number 1 4 M means group - ch ₂ - , -CO- or -CH ₂ CO- and R° means group - ch ₂ well ^ ² , -CH ₃ or -N0 _? • · · • ffffff · • ff ff » ♦ · · 10. A pharmaceutical composition, characterized in that it contains at least one cascade polymer complex according to claim 1, optionally with oaths customary in galenics. 11. Use of at least one polymer complex according to claim 1 for the production of means for NMR diagnostics or X-ray diagnostics for both conventional methods and computed tomography. 12. Use of cascade polymer complexes according to claim 1 for the differentiation of incipient and malignant tumors in body regions without a blood-brain barrier. 13. The method of producing complex cascade polymers according to one of claims 1 to 9, characterized in that compounds of the general formula I * (I') are allowed to react, in which the nitrogen-containing cascade core denotes the basal multiples and, X and Y denote, independently of each other, a direct link or a cascading reproductive unit of the reproductive multiplicity x or y, Z and V independently denote the cascading reproductive unit of reproductive multiplicity z , respectively w , M 9 9 9 9 99 99 • 9 · 99 99 a denotes the number 2 to 12 , x, y, z and w independently denote the number 1 to 4 and β denotes the binding site of the terminal NH-groups of the last generation of the reproductive unit V , with the restriction that at least two reproductive units are different and that multiplicities apply to the product 16^a.x.y.z.w^64, with a complex or complexing substance K' of the general formula I ’ A or I ’ B R ¹ OOC-R ² HC N —CH ₃ —CH ₂ —N And ι! i ch, ch, ch ₂ ch ₂ N—CH ₂ —CH ₂ —N (PA) CHR -COOR chr ² <oor ¹ ·· ·* * · · « »· « · • * · R ROOC-HjC [ N—CH ₂ CH _; r'ooc-h„c and. CH-OOCHj-COOR ¹ N-CH,CK ₂ -N CH,-C OOR ^: while R denotes, independently of each other, a hydrogen atom, the equivalent of a metal ion of serial number 20 to 29, 39, 42 to 44 or 57 to 83, or an acid protecting group, R ² denotes a hydrogen atom, a methyl group or an ethyl group, which is optionally substituted by one or two hydroxy groups or one carboxy group, R ⁴ R ² R ³ denotes the group -CH-CO-NU^-T', whereby R ⁴ denotes a straight or branched, saturated or unsaturated alkyl chain, which is optionally interrupted by 1 to 10 oxygen atoms, one phenylene or one phenylenoxy group and/or is optionally substituted by 1 to 5 hydroxy groups, 1 to 3 carboxy groups or one phenyl group, U ⁶ means straight or branched, saturated or 100 • » 4 · · · 4 4·« • · · · · · * 4 4 · 4 _{A e} 4 4 4 444 · 4 · 4· *4<4 4· 444 44 «« unsaturated alkylene group with 1 to 20 carbon atoms, optionally containing 1 to 5 imino groups, 1 to 3 phenylene groups, 1 to 3 phenoxy groups, 1 to 3 phenyleneimino groups, 1 to 5 amido groups, 1 to 2 hydrazide groups, 1 to 5 carbonyl groups, 1 to 5 ethyleneoxy groups, 1 urea group, 1 thiourea group, 1 to 2 carboxyalkylimino groups, 1 to 2 ester groups, 1 to 10 oxygen atoms, 1 to 5 sulfur atoms and/or 1 to 5 nitrogen atoms and/or optionally substituted by 1 to 5 hydroxy groups, 1 to 2 mercapto groups, 1 to 5 oxo groups, 1 to 5 thioxo groups, 1 to 3 carboxy groups, 1 to 5 carboxyalkyl groups, 1 to 5 ester groups and/or 1 to 3 amino groups, whereby the phenylene groups optionally present may be substituted by 1 to 2 carboxy groups, 1 to 2 sulfone groups or 1 to 2 hydroxy groups, and T' denotes the group -C*0, -COOH, -N=C=O or -N=C=S, C'0 denotes a hydrogen atom or a group and denotes an activated carboxyl group, with the restriction that if K' denotes a complex, at least two (for divalent metals) or three (for trivalent metals) of the substituents denote the equivalent of a metal ion of the above-mentioned elements and that there are additional carboxyl groups as needed in the form of their salts with inorganic and/or organic bases, amino acids or amino amides• * • « 101 » · 44 • ·· · , « · I ·· ·· acids, or the protective groups present are cleaved off, the cascade polymers thus obtained - if K' denotes a complexing substance - are allowed to react in a known manner with at least one metal oxide or metal salts of the element with serial number 20 to 29, 39, 42, 44 or 57 to 83 and, optionally, then in the complexes of cascade polymers obtained in this way, the acidic hydrogen atoms still present are completely or partially substituted by cations of inorganic and/or organic bases, amino acids or amino acid amides and optionally, the terminal amino groups still present are acylated as needed - before or after complexation with the metal. 14, Compounds of general formula I'A 0OC-R ² HC N —CH ₂ —CH ₂ — N ch ₂ N—CH ₂ —CH ₂ —n 3' R (ΙΆ) CHR -COOR chr ² <oor in which R ¹ ' independently denotes a hydrogen atom, the equivalent of a metal ion of serial number 20 to 29, 39, 42 to 44 102 • * · ** * · · 9 ··· 9 ·· · 9 9 ··· 9 9 «9·· 99 9 · 9 99 «· or 57 to 83 , or an acid protecting group, R ² denotes a hydrogen atom, a methyl group or an ethyl group, which is optionally substituted by one or two hydroxy groups or one carboxy group, R ⁴ R ² R^ denotes the group -CH-CO-N-U^-T', while R ⁴ denotes a straight or branched, saturated or unsaturated alkyl chain, which is optionally interrupted by 1 to 10 oxygen atoms, one phenylene or one phenyleneoxy group and/or is optionally substituted by 1 to 5 hydroxy groups, 1 to 3 carboxy groups or one phenyl group, means a straight or branched, saturated or unsaturated alkylene group with 1 to 20 carbon atoms, optionally containing 1 to 5 imino groups, 1 to 3 phenylene groups, 1 to 3 phenoxy groups, 1 to 3 phenyleneimino groups, 1 to 5 amido groups, 1 to 2 hydrazide groups, 1 to 5 carbonyl groups, 1 to 5 ethyleneoxy groups, 1 urea group, 1 thiourea group, 1 to 2 carboxyalkylimino groups, 1 to 2 ester groups, 1 to 10 oxygen atoms, 1 to 5 sulfur atoms and/or 1 to 5 nitrogen atoms and /or optionally substituted with 1 to 5 hydroxy groups, 1 to 2 mercapto groups, 1 to 5 oxo103 groups, 1 to 5 thioxo groups, 1 to 3 carboxy groups, 1 to 5 carboxyalkyl groups, 1 to 5 ester groups and/or 1 to 3 amino groups, wherein the phenylene groups optionally present may be substituted with 1 to 2 carboxy groups, 1 to 2 sulfone groups or 1 to 2 hydroxy groups, and T' denotes the group -C*O, -COOH, -N=C=0 or -N=C=S and C*0 denotes an activated carboxyl group. 15. The method of manufacturing pharmaceutical preparations according to claim 10, characterized in that the cascade polymer complexes, dissolved or suspended in water or in a physiological salt solution, are converted with the possible help of usual additives in a galenical into a form suitable for enteral or parenteral application.