WO1999032154A1

WO1999032154A1 - Dendritic polymer-saccharide conjugate, pharmaceuticals containing said conjugate, method for its production and use thereof

Info

Publication number: WO1999032154A1
Application number: PCT/EP1998/007927
Authority: WO
Inventors: Peter Mareski; Johannes Platzek; Bernd Radüchel; Hanns-Joachim Weinmann; Dietmar Berndorff; Bernd Misselwitz
Original assignee: Schering Aktiengesellschaft
Priority date: 1997-12-18
Filing date: 1998-12-09
Publication date: 1999-07-01
Also published as: EP1037672A1; DE19758105A1; AU2268099A

Abstract

The invention concerns conjugates consisting of dendritic polymers amino groups whose branches are produced from vinyl cyanide units, metal ion(s) containing a signal-emitting group or groups and monosaccharides or oligosaccharides and optionally cations of organic and/or inorganic bases, amino acids or amino acid amides. Said conjugates are valuable compounds in diagnosis and therapy.

Description

Dendritic polymer-saccharide conjugates, pharmaceutical compositions containing them, processes for their preparation and their

use

The invention relates to the subject matter characterized in the claims, that is to say conjugates consisting of an amino group-containing dendritic polymer, the branches of which are made from vinyl cyanide units, signaling group (s) containing metal ion (s) and mono- or oligosaccharides and optionally cations of inorganic and / or organic bases, amino acids or amino acid amides, pharmaceutical compositions containing these compounds, their use in diagnostics and methods for producing these compounds and compositions.

Lymph node metastases become malignant in approximately 50-70% of patients

Tumors found (Elke, M. lymph node diseases. In: W. Frommhold, H.- S. Stender and P. Thurn (eds.), Radiological Diagnostics in Clinic and Practice, 7th edition, Vol. IV, pp. 475-512, Stuttgart-New York: Georg Thieme Verlag, 1984). The involvement of lymph nodes in a malignant disease has a significant influence on the therapeutic measures to be initiated. The earliest and most accurate diagnosis of lymph node involvement is therefore of the utmost importance for the patient.

Contrast-assisted lymphography has so far provided only unsatisfactory results in medical practice. X-ray examinations with iodized oils, e.g. with iodinated fatty acid esters of poppy oil, show good storage capacities in the lymph nodes and a useful contrast effect on the one hand, but on the other hand they are due to their known side effects (Keinert, K., Köhler, K., and Platzbecker, H. , Complications and contraindications In: M. Lüning, M. Wiljasalo, and H. Weissleder (eds.), Lymphography for malignant tumors, pp. 40-50. Stuttgart: Georg Thieme Verlag, 1976) neither general nor problem-free. They can promote lesions, particularly if they are applied too quickly, with regard to the vulnerability of the lymphatic vessels. A number of disadvantages are due to the oil character of these contrast agents. Since the oil is only suspended in the blood, the size of the oil droplets cannot be determined in advance. If the oil droplets are too large, microembolisms, for example in the lungs, are often avoidable. Since it is hardly possible to absorb the large oil droplets from the interstitial space into the lymph capillaries, the lymphatic vessels and lymph nodes can only be displayed in exceptional cases, for example after subcutaneous or intra-parenchymal injection.

The latter disadvantage can only be overcome by endolymphatic application, although prior dye marking and surgical exposure of the lymphatic vessels under local anesthesia must also be accepted without complications. Another disadvantage is that the contrast agents based on iodized oils are only excreted very slowly after application. Depending on the form of preparation, the elimination time is weeks to months.

By emulsifying the iodized oils, attempts have been made to overcome the disadvantages shown. This has resulted in a reduction in viscosity and droplet size. In this way the capillary passage could be improved and the risk of embolization reduced if the emulsion was stable. However, these dividers could only be bought for other disadvantages. Examples include the effective reduction of the iodine content per ml of contrast medium and the associated inevitable loss of contrast, greater local toxicity at the lymph nodes, hepatotoxic effects and histologically detectable foreign body reactions, which are enhanced by the added emulsifiers.

All contrast media based on iodized oils are not very stable and can only be used to a limited extent due to the side effects shown. The derivatives of triiodinated 5-aminoisophthalic acid proposed as lymphographic agents in EP 0 022 056 have not met the expectations placed on them in medical practice because they diffuse out of the lymphatic system again after application. Due to their inadequate storage capacity in the lymph nodes, these aqueous preparation forms are at best only suitable for peripheral lymphography to a limited extent.

Water-soluble iodinated X-ray contrast media such as lotrolan (Isovist®) have only a limited area of application for the representation of the lymphatic system. By means of infusion into the interstitial space (intracutaneous, subcutaneous or intramuscular), only those in the catchment area of the infusion lymphatic vessels or nodes are shown in high contrast. The highly water-soluble, low-molecular substances then diffuse from the lymphatic vessels into the interstitium. This disadvantage cannot be compensated for by the sensitive method of computed tomography.

In nuclear medicine, radioactive ^ ^m Tc tracers are used for indirect lymph node imaging. In addition to the limited spatial resolution, the disadvantages also apply here that only the lymph channels supplying the injection site are shown.

Superparamagnetic iron oxide particles are also suitable as lymphographics for nuclear magnetic resonance imaging, e.g. AMI-227, have been proposed (Guimaraes, R., Clement, O., Bittoun, J., Carnot, f., And Frija, G. MR lymphography with superparamagnetic iron nanoparticles in rats: pathologic basis for contrast enhancement. Am J. Roentgenol., 162, 201-207, 1994). However, the examination results obtained on the animal so far indicate unsatisfactory imaging despite the high dosage of the particles.

Various compounds proposed as lymphographics (Hanka, L. et al., Radiology 1996, 198, 365-370) show such animal specificity (rat vs. guinea pig and rabbit) that their further development had to be stopped.

There was therefore still the task of providing compounds which accumulate in the lymphatic tissue after parenteral administration and do not have the disadvantages mentioned above, e.g. have a more favorable safety margin between the diagnostic and lethal dose ("margin of safety") compared to the known structures. This object is achieved by the present invention.

It has been found that the conjugates according to the invention consist of a dendritic polymer containing amino groups, the branches of which are made from vinyl cyanide units, signaling group (s) containing metal ion (s) and mono- or oligosaccharides and optionally cations of inorganic and / or organic bases, amino acids or Amino acid amides are surprisingly particularly suitable for the production of NMR diagnostics, in particular for lymphography.

Surprisingly, they have the following advantages:

Enrichment in the lymphatic system sufficient for good imaging, high tolerance (LD50), short elimination time (more than 98% of them are usually excreted within 14 days), their high relaxivity allows them to be reduced Dosage to apply, they often even allow a morphological differentiation of the

Lymph node tissue, they show no deviating animal specificities.

The conjugates according to the invention can be described by the general formula I:

P (K) _m (LZ) _n (I),

wherein

P for a k amino groups - of which (n + m) are each reduced by one hydrogen atom - containing dendritic polymer, the branches of which are made from vinyl cyanide units, where (n + m) <k, n and m in each case for the integers 1 - 149, k for the numbers 12 to 150,

K stands for a signaling group containing metal ion (s), L for a linker and Z for a mono- or oligosaccharide reduced by one hydroxyl group.

The polymer which stands for P and contains k amino groups, of which (n + m) are each reduced by one hydrogen atom, is a dendrimer prepared from vinyl group units, as described in the patent specifications WO 93/14147, WO 93/12073, WO 95/02008, WO 95/20619, WO 96/02588, EP 684044, EP 672703 and US Pat. No. 5,530,092 and which DSM polyamines are called, the repeating units being 3-amino-propylene or 3-amino-2-methyl-propylene groups [cf. see also pages 34 and 35 for the corresponding formulas]. The sum (n + m) of the signaling groups (K) and of the mono- or polysaccharides (Z) bound by L, each reduced by one hydroxyl group, is preferably equal to the number k of the amino groups contained in the polymer, ie. There is a complete occupation of the amino groups contained in P. The polymers preferably contain 32 or 64 amino groups, of which, for example, 3, 4, 6, 7, 8, 11, 13, 14, 16, 18, 19, 21, 47 on signaling groups K and, for example, 9, 8, 18, 17 , 16, 13, 11, 10, 20, 6, 29, 27, 33 are bound to mono- or polysaccharides Z which are each bound via L and reduced by one hydroxyl group.

If the amino groups are understaffed, the sum (n + m) is preferably> k - 4.

The signaling group K is a chelate complex consisting of a radical of the general formef II, III, IV, V or VI.

R ¹ OOC

NNN (III), R ¹ OOC "1 ^ COOR ¹

R ² ^ C = O

wherein

R1 independently of one another is a hydrogen atom or a metal ion equivalent of the elements with a number of 20-32, 37-39, 42-44, 49 or 57-83,

R ^{2 is} a hydrogen atom, a straight-chain or branched C 1 -C 7 -alkyl radical, a phenyl or benzyl radical,

V group

With

0 in the meaning of the numbers 0-10, p and I in each case in the meaning of the numbers 0 or 1, with the proviso that p only stands for the number 1 if I means the number 1, T1 a -NHCS- or -CO group,

U is a -CHR ³ -CONR ³ -M ¹ or -CH2-CH (OH) -M ² group with R ³ independently of one another in the meaning of R2 or the group -CH2- (CH2) ₀ -

COOH and Ml and M ² each have the meaning of a phenylene radical or a straight-chain, branched, saturated or unsaturated C-1 -C20-alkylene chain which is optionally substituted by 1 -5 (CH2) ₀ -COOH, 1 -5OR ² radicals or 1 -8 oxygen atoms, 1 -2-NH-, 1 -2-C (= NH) -, 1 -5- CONR ^3- , 1 -5-NR ³ CO-, 1 -2 phenylene- or 1 -2 Contains phenyleneoxy groups, with the proviso that at least two of the radicals R1 are metal ion equivalents of the elements of the above-mentioned atomic numbers.

If the compound according to the invention is intended for use in NMR diagnostics, the metal ion of the signaling group must be paramagnetic. These are in particular the divalent and trivalent ions of the elements of atomic numbers 21-29, 42, 44 and 58-70. Suitable ions are, for example, chromium (III) -, iron (II) -, cobalt (II) -, nickel ( ll) -, copper (ll) -, Praseodymium (III), Neodymium (III), Samarium (III) and Ytterbium (III) ion. Because of their strong magnetic moment, gadolinium (III), terbium (III), dysρrosium (III), holmium (III), erbium (III), iron (III) and manganese (II) are particularly preferred. -ions.

The metal ion must be radioactive for the use of the compounds according to the invention in nuclear medicine. For example, radioisotopes of the elements copper, cobalt, gallium, germanium, yttrium, strontium, technetium, indium, ytterbium, gadolinium, samarium, iridium, rhenium and bismuth are suitable; technetium, gallium, indium and rhenium are preferred.

If the compound according to the invention is intended for use in X-ray diagnosis, the metal ion is preferably derived from an element of a higher atomic number in order to achieve sufficient absorption of the X-rays. It has been found that diagnostic agents which contain a physiologically compatible complex salt with metal ions of elements of atomic numbers 25 and 26 and 57-83 are suitable for this purpose.

Manganese (II) -, iron (II) -, iron (III) -, praseodymium (III) -, neodymium (III) -, samarium (III) -, gadolinium (III) -, ytterbium (III) are preferred - or bismuth (III) ions, especially dysprosium (III) ions.

Acidic hydrogen atoms which may be present in R1, that is to say those which have not been substituted by the central ion, can, if appropriate, be replaced in whole or in part by cations of inorganic and / or organic bases or amino acids or amino acid amides.

Suitable inorganic cations are, for example, lithium ion, potassium ion, calcium ion and in particular sodium ion. Suitable cations of organic bases include those of primary, secondary or tertiary amines, such as, for example, ethanolamine, diethanolamine, morpholine, glucamine, N, N-dimethylglucamine and in particular N-methylglucamine. Suitable cations of amino acids are, for example, those of lysine, arginine and ornithine and the amides of otherwise acidic or neutral amino acids. The compounds according to the invention, which have a molecular weight of 5,000-200,000 D, preferably 15,000-60,000 D, have the desired properties described at the outset. They contain the large number of metal ions required for their use which are stably bound in the complex.

They accumulate in the lymphatic system, allow statements about the perfusion of tissues, give the possibility to determine the blood volume in tissues and selectively shorten the relaxation times or densities of the blood. Such physiological information cannot be obtained through the use of extracellular contrast agents, e.g. Gd-DTPA [Magnevist®]. From these points of view, the areas of application for the modern imaging methods magnetic resonance imaging and computed tomography also result: more specific diagnosis of metastatic, especially malignant tumors and early therapy control with cytostatic, anti-inflammatory or vasodilative therapy.

The saccharide-polymer complexes according to the invention are outstandingly suitable for the interstitial and especially for the i.v. Lymphography.

As further advantages over extracellular contrast agents, e.g. Gd-DTPA [Magnevist®], the higher effectiveness as a contrast agent for magnetic resonance imaging (higher relaxivity) must be emphasized, which leads to a significant reduction in the diagnostically necessary dose. At the same time, the contrast media according to the invention can be formulated as solutions isoosmolar to the blood and thereby reduce the osmotic load on the body, which is reflected in a reduced toxicity of the substance (higher toxic threshold). Lower doses and higher toxic thresholds lead to a significant increase in the safety of contrast medium applications in modern imaging processes.

Another advantage of the present invention is that complexes with hydrophilic or lipophilic, macrocyclic or open-chain, low molecular or high molecular weight ligands have now become accessible. This enables the tolerance and pharmacokinetics of these polymer complexes to be controlled by chemical substitution. Preferred substituents for R ² are the hydrogen atom, the methyl, isopropyl, phenyl and benzyl group. Examples of preferred groups standing for V are the CH2C6H4, CH2-O-C6H4, (CH2) 4, (CH ₂ ) 6 and (CH2) ι o groups, the C6H4 group being attached to T ^" ! is bound.

Preferred substituents for R ³ are the hydrogen atom, the methyl, CH ₂ COOH and (CH ₂ ) 2COOH group.

Groups M ^"1 may be mentioned as examples

α- (CH ₂ ) ι _ιo-ß, -CH (CH ₃ ) -ß,

α-CH [CH ₂ CH (CH ₃ ) ₂ ] -ß, α-CH- (C ₆ H ₅ ) -ß

α-CH ₂ -CH ₂ -O ^ r-CH ₂ - ß, α-CH ₂ CH- ß _{> α} - (CH ₂ CONH) ^ - CH ₂ - ß,

CH ₃ α-CH ₂ -CH ₂ -O-CH ₂ -ß, α- (CH2) 2-3- (OCH ₂ CH ₂ ) ι _5-O- (CH ₂ ) 2-3-ß,

where α indicates the linkage to the rest-CONR ³ and ß the linkage to T ^" !

The groups are preferred:

α CH ₂ - ß, α (CH ₂ ) ₂ ß, α (/ ß, CH ₂ COOH

α CH-CH ₂ - ß, _α -CH ₂ -CH ₂ -O-CH ₂ -ß, α- (CH ₂ ) ι.2-CO-N-CH ₂ -ß,

CH ₂ COOH

-CH2-CH ₂ -CO-NH (CH ₂ ) 6-ß, α- (CH ₂ ) 5-ß,

Examples of groups M ² are:

where α indicates the link to the rest-CH (OH) - and ß the link to T ¹ .

The groups are preferred:

α -CH ₂ -ß,

CH ₂ COOH

α -CH ₂ -NHCO- (CH ₂ ) _2.3 -ß

^α - CH - OP ß, ^α - CH ₂ -NHCO-CH ₂ -O-CH ₂ - ß, a (CH ₂ ) _g - ß The group is particularly preferred:

Groups Z for the monosaccharides are the hexoses, pentoses and N-acetyl-neuraminic acid, each reduced by one hydroxyl group, which are each in the pyranose or furanose form, and their derivatives. Preferred are hexoses which are in the pyranose form, such as, for example, D-galactose, D-mannose, D-glucose, L-fucose, galactose-6-sulfate, mannose-6-sulfate, D-glucosamine. 6-sulfate, D-glucosamine-3,4,6-trisulfate and N-acetyl-glucosamine. Examples of preferred monosaccharides are N-acetyl-2-amino-2-deoxy-D-glucose, -D-galactose and D-mannose, 6-deoxy-L-galactose and N-acetyl-neuraminic acid. If Z stands for oligosaccharides, these can be linear or branched from the monosaccharides mentioned above, and the linkage can be terminal or within the chain. Examples of oligosaccharides are: sialyl-Lewis ^x (see, for example, H. Ohmoto et al, J. Med. Chem. 1996, 39, 1339); Deoxy-sialyl-Lewis ^x

(see, for example, W. Stahl et al, Angew. Chemie 1994, 106, 2186); Sialyl-Le ^x (e.g. W.

Stahl, J. prak. Chemistry 1995, 337, 441); Gentiobiose (Fluka Chemie AG,

Switzerland); Lactose; Maltose; Lactulose (Fluka Chemie AG, Switzerland); LactNAc; α N, N ', N "-triacetylchitothose; Gal (1 ^~ * 4) Gal, N, N'-diacetylchitobiose; N, N', N", N '"- tetraacetylchitotetraose; maltotriose; digalacturonic acid; trigalacturonic acid (carbohydrates International AB, Sweden).

The polymer is preferably bound (via the linker L) to the mono- or polysaccharides via the C-2 position, particularly preferably via the C-1 position of the sugars.

The linker L, via which the mono- or oligosacchaides reduced by one hydroxyl group are bound to P, is described by the group T ² -M -X, where M ^{3 stands} for a direct bond or the one given for M ^" ! T ^{2 is} an -NHCS-, -C (= NH) -, -CH2- or CO group and binds to P, X stands for NH, CO, O and S and binds to Z. The groups M ³ are, for example

γ- (CH ₂ ) ii ₅ - δ,

γ- (CH ₂ CH ₂ O) ₁ .5-CH2-δ

γ-CH ₂ - (C = NH) -δ, γ-C (= NH) -NH- (CH ₂ ) 3-δ,

γ-NHCO- (CH ₂ ) 2-δ, γ-CH ₂ CH2-O-CH ₂ CH2-δ, γ-CH ₂ CH2-S-CH ₂ CH2-δ,

γ -OCH ₂ - δ, γ -NH-CH (CH ₂ - // V_J

called, where γ indicates the junction at X and δ the junction at T ² .

M ³ preferably stands for a direct bond and for the groups

T— (CH ₂ CH ₂ O) ₁ . ₄ - CH ₂ - (CH ₂ ) _0.1 - _δ , γ_ (CH ₂ ) ₂ -CONH- (CH ₂ ) ₆ -δ _ι

CH (OH) CH ₂ OH I y - CH-CH (OH) -CH (OH) -δ, γ- (CH2) 2-CO- (CH ₂ ) 3-δ The conjugates of the general formula according to the invention are prepared in a manner known per se that

a) a polymer complex (former) conjugate of the general formula VII

P (K ') m (VII), where K' is a radical of the general formula II ', III', IV, V or VI '

OC - \ / - \ / - \ ^ - COOR

<■■■ "> -

wherein

Rl 'independently of one another for a hydrogen atom, an acid protecting group or a metal ion equivalent of the elements of atomic numbers 20-29,

39, 42, 44 or 57-83 stands and

R ² , V, o, p, T and U have the meaning given above, with n 'mono- or oligosaccharides of the general formula VIII

L ^* -2 (VIII),

where n '= 1-3 n, Z 'has the meaning given for Z, but the carboxy, amino and hydroxyl groups which may be present in Z are optionally protected,

L * for a group T ² '-M ³ ' -X with T ² 'in the meaning of -NCS, -C (= NH) OCH3, - CHO, a -CO-Fg group where

-O-CO-R ^ with R ^ meaning a straight-chain or branched C <| -C7 alkyl chain,

M ³ 'has the meaning given for M ³ , but the carboxy groups which may be present in M ³ are optionally protected, and X represents NH, CO, O and S, or b) a polymer saccharide conjugate of the general formula IX

P (L'-Z ') _n (IX),

where L 'has the meaning given for L, but the carboxy groups which may be present in L are optionally protected and P, Z' and n have the meaning given above, with m 'complexes or complexing agents K *, where m' = 1-3 m is and K ^* for compounds of the general formulas

Fg— OC - / - ^ - x ^ COOR ^{1 '} NNN

R OOC ^ I ^ - COOR ^{1 '}

COOR ¹ _{(| r) ι}

Fg (III "),

(VI ")

wherein

R1 ', R ² , V, U and Fg have the meaning given above, and T ¹ ' stands for -NCS, -CO-Fg, -C (= NH) OCH ₃ or -CHO or

c) a polymer of the general formula X

P- (H) _k (X),

wherein

P and k have the meanings given above, in a one-pot reaction with n 'mono- or oligosaccharides of the general formula VIII

L ^* -Z '(VIII),

wherein n ', L ^* and Z' have the meanings given above, with m 'complexes or complexing agents K ^* , where K ^* for compounds of the general formula

Fg— OC- / —χ - \ ^ -COOR ¹ NNN

R ^r OOC ^ ^ ^ COOR ¹

COOR

(II "),

R ^{1 '} θOC- \ / \ / \ _/ -COOR ¹

N N N

R'OOC- ⁷ "l ^ -COOR ^{1 '}

R 'C = O i Fg (III "),

(VI ")

wherein m ', R ¹ ', R ² , V, T1 ', and Fg have the meaning given above, the metal ion introduction which may be necessary being carried out before or after the splitting off of the protective groups which may be necessary, with the proviso that if Rl 'stands for protective groups, these must be split off before the metal ion is introduced.

If R1 'represents an acid protecting group or if the caboxy groups contained in L', M ³ 'and Z' are protected, lower alkyl, aryl and aralkyl groups, for example the methyl, ethyl, propyl, butyl, Phenyl, benzyl, diphenylmethyl, triphenylmethyl, bis (p-nitrophenyl) methyl group, and trialkylsilyl groups in question.

If desired, the protective groups are split off by the processes known to the person skilled in the art (see, for example, E. Wünsch, Methods of Org. Chemistry, Houben-Weyl, Vol XV / 1st 4th Edition 1974, p. 315), for example by hydrolysis, hydrogenolysis, alkaline saponification of the esters with alkali in aqueous-alcoholic solution at temperatures from 0 ° C to 50 ° C, acid saponification with mineral acids or in the case of tert-butyl esters with the help of trifluoroacetic acid. The hydroxyl protective groups are, for example, the benzyl, 4-methoxybenzyl, 4-nitrobenzyl, tπtyl, diphenylmethyl, tπmethylsilyl, dimethyl-t-butylsilyl-diphenyl-t-butysilyl group as a protective group for the 1 -OH group of the mono- and / or oligosaccharides is also suitable for the methyl group

The hydroxyl groups can also, for example, as THP ether, alkyl ether, α-alkoxyethyl ether, MEM and MOM ether or as an ester with aromatic or aliphatic carboxylic acids, such as acetic acid and chloroacetic acid, benzoic acid, levulinic acid or 2 Chloroacetoxymethyl (or ethyl) benzoic acid (GIT Fachz Lab 1996, 46) are present. In the case of polyols, the hydroxyl groups can also be protected in the form of ketals with, for example, acetone, acetaldehyde, cyclohexanone or benzaldehyde

If carboxyl groups are present at the same time, hydroxyl groups can also be protected by intramolecular esterification to give the corresponding lactones

The hydroxyl protective groups can be released using the literature methods known to the person skilled in the art, for example by hydrogenolysis, reductive cleavage with lithium / ammonia, acid treatment of the ethers and ketals or alkali treatment of the esters (see, for example, “Protective Groups in Organic Synthesis”, TW Greene, John Wiley and Sons 1981)

The amine protecting groups are the benzyloxycarbonyl, tertiary-butoxycarbonyl, trifluoroacetyl, fluorenylmethoxycarbonyl, benzyl, formyl, 4-methoxybenzyl, 2,2,2-trιchlorethoxycarbonyl, phthaloyl, 1,2- Oxazoline, tosyl, dithiasuccinoyl .AIIyloxycabonyl, sulfate, pent-4-encarbonyl, 2-chloroacetoxymethyl (or ethyl) benzoyl, tetrachlorophthaloyl, alkyloxycarbonyl groups [Th W Greene, PGM Wuts, Protective Groups in Organic Syntheses, 2nd ed, John Wiley and Sons (1991), S 309 - 385, E Meinjohanns et al, J Chem Soc Pekin Trans 1, 1995, 405, U Ellensik et al, Carbohydrate Research 280, 1996, 251, R Madsen et al, J Org Chem 60, 1995, 7920, R R. Schmidt, Tetrahedron Letters 1995, 5343]

The reaction indicated under a) is carried out according to the methods familiar to the person skilled in the art, such as, for example, in P Erbacher et al, Bioconjugate Chem 1995, 6, 401, G Molema et al, J Med Chem 1991, 34, 1137 JL Montero et al, Bull Soc. Chim. France, 1994, 1_31_, 854; P. Midoux et al, Nucleic Acids Research 1993, 21, 871; Andersson, M. et al., Bioconjugate Chem. 1993, 4, 246; R. Roy et al, Tetrahedron Letters 1995, 36 4377; UK Saha et al, J. Chem. Soc. Chem. Comm. 1995, 2571; CR McBroom et al, Methods Enzymol. 1972, 28, 212 are described. It is in solvents such as water, methylene chloride, acetonitrile, chloroform, DMSO, pyridine, ethanol / water, ethanol / acetonitrile, dioxane, DMF, THF, lower alcohols, tetramethylurea, N-methylpyrrolidone, polyethylene glycols, 12-dimethoxyethane, dimethylacetamide , 1, 2-dichloroethane or - if possible - their mixtures with water at temperatures from -10 ° C to 100 ° C, preferably 0 to 50 ° C, particularly preferably at room temperature within 5 minutes to 72 hours, preferably 1 to 24 Hours, particularly preferably 1 to 14 hours, optionally with the addition of an organic or inorganic base, such as aromatic or aliphatic amines, alkali or alkaline earth metal hydroxides, carbonates or hydrogen carbonates and quaternary ammonium hydroxides. Examples include triethylamine, di-isopropyl-N-ethylamine (Hünig base), N-methylmorpholine, tributylamine, tetramethylethylenediamine, pyridine, lutedine, 2,4,6-trimethylpyridine, 4-dimethylaminopyridine, N-methylimidazole, tetramethylguanidine, DBU, Lithium, sodium, potassium, calcium, magnesium, barium hydroxide, carbonate, and hydrogen carbonate. The reaction can also be carried out in the manner known to those skilled in the art

Buffer solutions, preferably at pH 8 to 11, particularly preferably at pH 8.5 to 9. The pH value is preferably maintained using a pH state. If T ² 'stands for a CHO group, coupling is carried out by reductive amination with the aid of a reducing agent according to methods known to the person skilled in the art (J.-P. Sabri et al, Tetrahedron Letters 1994, 35, 1181; MD Bomaπn et al, J Org. Chem. 1995, 60, 5995; S. Bhattacharyya et al, J. Chem. Soz. Pekin Trans. I 1994, 1; OS Artyushin et al., Ser. Khim 1991, 9, 2154; RF Borch et al , JACS 1971, 93 2897). Examples of reducing agents used are sodium borohydride, sodium cyanoborohydride, lithium borohydride, calcium borohydride, pyridine-BH3, lithium aluminum hydride and zinc borohydride. Catalytic hydrogenation on, for example, palladium or nickel catalyst is also suitable.

If Fg stands for an OH group, an in-situ activation with the coupling reagents known to the person skilled in the art, such as e.g. DCCI, EEDQ,

Staab reagent, BOP, PyBOP, TBTU, TDBTU, HBTU (see e.g. Fournic-Zaluski et al, J Med Chem 1996, 39, 2596, Houben-Weyl, Volume XV / 2, Part II, 1974, YM Angell et al, Tetrahedron Letters 1994, 35, 5981, LA Carpino et at, J Chem Soc Commun 1994, 201 , HO Kim et al, Tetrahedron Letters 1995, 36, 6013, D Papaioannou et al, Tetrahedron Letters, 1995, 36, 5187, G Stemple et al, Bioorg Med Letters 1996, 6, 55

The reaction of the polymer saccharide conjugate of the general formula IX indicated under b) with the complexes or complexing agents K ^{* of} the general formula II "to VII" and II'a is also carried out in a manner known per se, as described, for example, in US Pat. No. 5,135,737 , H Takalo et al, Bioconjugate Chem 1994, 5, 278, EP 0430863, EP 0331616, WO 96/01655 EP 0271 180, US-5,364,613, WO 95/17451 and WO 96/02669 It is in solvents such as water, Methylenchloπd, Acetonitnl, chloroform, DMS0, pyridine, ethanol / water, ethanol / Acetonitπl, dioxane, DMF, THF, lower alcohols, tetramethylurea, N-methylpyrrolidone, polyethylene glycols, 12-dimethoxyethane, dimethylacetamide, 1, or 2-dimethylchloride possible - Their mixtures with water at temperatures from -10 ° C to 100 ° C, preferably 0 to 50 ° C, particularly preferably at room temperatures within 5 minutes to 72 hours, preferably 1 to 24 hours, particularly preferably 1 to 14 hours, if appropriate with the addition of an organic n or inorganic base, such as aromatic or aliphatic amines, alkali or alkaline earth metal hydroxides, carbonates or hydrogen carbonates and quaternary ammonium hydroxides, examples which may be mentioned are methylamine, di-isopropyl-N-ethylamine (Hunig base), N-methylmorpholine, Tπbutylamine, tetramethylethylenediamine, pyridine, lutedin, 2,4,6-Tπmethylpyπdιn, 4-Dιmethylamιnopyrιdιn, N-methyl midazole, tetramethylguanidine, DBU, lithium, sodium, potassium, calcium, magnesium, tree hydride , -carbonate, -hydrogencarbonate The reaction can also be carried out in the buffer solutions known to the person skilled in the art, preferably at pH 8 to 11, particularly preferably at pH 8.5 to 9. The pH is preferably maintained using a pH state of T. ² 'for a CHO group, a coupling is carried out by reductive amination with the aid of a reducing agent according to methods known to the person skilled in the art (J-P Sabri et al, Tetrahedron Letters 1994, 35, 1181, MD Bomann et al, 7 Org Chem 1995, 60, 5995, S Bhattacharyya et al, J Chem Soz Pekin Trans I 1994, 1, OS Artyushin et al, Ser Khim 1991, 9, 2154, RF Borch et al, JACS 1971, 93 2897) The reducing agents used are, for example, sodium borohydride, sodium cyanoboron hydride, lithium borohydride, calcium borohydride, pyridine-BH3, lithium aluminum hydride and zinc borohydride. Catalytic hydrogenation is also suitable.

The reactions indicated under a) and b) are preferably carried out with an excess of the activated saccharide of the formula VIII or the activated complex or complexing agent K ^*, preferably with a 1.5 to 3-fold excess.

The dropping reaction indicated under c) takes place under the reaction conditions listed under a) and b) by using a polymer of the general

Formula X implemented with a mixture of m '= 1 - 3 m K ^* and n' = 1 - 3 n L ^* - Z '. The occupancy ratio of the amino groups contained in the polymer is controlled by the ratio n ': m'.

The metal ion introduction which is necessary after the above-mentioned reactions a) - c) or has already been carried out in K 'or K * takes place in the manner described, for example, has been disclosed in German Offenlegungsschrift 34 01 052 by using the metal oxide or a metal salt (for example the nitrate, acetate, carbonate, chloride or sulfate) of the element of atomic numbers 20-32, 37-39, 42-44, 49 , 57 - 83 dissolved or suspended in water and / or a lower alcohol (such as methanol, ethanol or isopropanol) and reacted with the solution or suspension of the equivalent amount of the complex-forming ligand and then, if desired, acidic hydrogen atoms of the acid groups present by cations of inorganic and / or organic bases, amino acids or amino acid amides.

The neutralization takes place with the aid of inorganic bases (for example hydroxides, carbonates or bicarbonates) of, for example, sodium, potassium, lithium, magnesium or calcium and / or organic bases such as, inter alia, primary, secondary and tertiary amines, for example ethanolamine, morpholine, Glucamine, N-methyl- 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, for example, hippuric acid, glycine acetamide.

To prepare the complex compounds, for example, the acidic complex salts in aqueous solution or suspension can be added with as much of the desired base that the neutral point is reached. The solution obtained can then be evaporated to dryness in vacuo. It is often advantageous to remove the neutral salts formed by adding water-miscible solvents, e.g. to precipitate lower alcohols (methanol, ethanol, isopropanol and others), lower ketones (acetone and others), polar ethers (tetrahydrofuran, dioxane, 1, 2-dimethoxyethane and others) and thus to obtain crystals that are easy to isolate and easy to clean. It has proven to be particularly advantageous to add the desired base already during the complex formation of the reaction mixture and thereby to save one process step.

If the acidic complex compounds contain several free acidic groups, it is often expedient to prepare neutral mixed salts which contain both inorganic and organic cations as counterions.

This can be done, for example, by reacting the complex-forming ligand in aqueous suspension or solution with the oxide or salt of the element providing the central ion and half of the amount of an organic base required for neutralization, isolating the complex salt formed, cleaning it if desired and then to complete it Neutralization with the required amount of inorganic base. The order of base addition can also be reversed.

The polysaccharide conjugates obtained in this way are purified, if necessary after adjusting the pH, by adding an acid or base to pH 6 to 8, preferably about 7, preferably by ultrafiltration with membranes suitable pore size (e.g. Amicon-®XM30, Amicon®-YM10, Amicon®-YM3) or gel filtration on e.g. suitable Sephadex® gels.

In the case of neutral compounds, it is often advantageous to give the polymeric complexes via an anion exchanger, for example IRA 67 (OH ^" form) and, if appropriate, additionally via a cation exchanger, for example IRC 50 (H ⁺ form), in order to remove ionic components.

The preparation of the polymer-complex (former) conjugates of the general formula VII required as starting substances

P (K ') _m (VII)

and the polymer saccharide conjugates of the general formula IX

P (L '- Z') _n (IX)

is carried out in a manner known per se by using a polymer of the general formula X

P- (H) _k (X) with m 'complexes or complexing agents K ^{* of} the general formulas II "-VI" and II'a or with N' activated mono- or oligosaccharides of the general formula VIII

L ^* - Z '(VIII)

under the reaction conditions given under a) and b) above.

The polymers of the general formula X can be used in the form of the free amines or in the form of their salts, for example as hydrochlorides, hydrobromides or hydrosulfates. They are known from the literature (e.g. WO 93/14147, WO 93/12073, WO 95/02008, WO 95/20619, WO 96/02588, EP 684044, EP 672703 and US-5,530,092).

The complexes or complexing agents K * used are known from the literature or can be obtained analogously to methods known from the literature:

II "and ll'a: see e.g. EP 263 059,

III ": see e.g. DE 19507822, DE 19580858, DE19507819

IV ": see e.g. WO 91/14459

V ": see e.g. US-5,053,503, WO 96/02669, WO 96/01655,

EP 0430863, EP 255471, US-5,277,895, EP 0232751,

U.S. 4,885,363

VI ": see e.g. U.S. 4,923,985

The mono- and oligosaccharides of the general formula VIII required as starting materials are likewise known from the literature or can be obtained analogously to methods known from the literature (see also experimental part): see, for example, EP 0 128097; AJ Jonas et al. Biochem. J. 1990, 268, 41; JP Patent 04211099; DH Buss et al, J. Chem. Soc. C. 1968, 1457; CM. Hilditch et al, J. Appl. Phycol. 1991, 3, 345; M. Monsigny et al, Biol. Cell 1984, 51, 187; EP Dubois et al, Bioorganic and Medical Chemistry Letters 1996, 6, 1387; CR McBroom et al, Methods Enzymol. 1972, 28, 212, M. Ponpipom et al, J. Med. Chem. 1981, 24, 1388; J. Haensler et al, Bioconjugate Chem. 1993, 4, 85; Y. Miura et al, Carbohydrate Research 1996, 289, 193; R. Roy et al. Tetrahedron Letters 1995, 36, 4377; W. Stahl et al, Angew. Chemistry 1994, 1_06, 2186; U. Sprengard et al, Angew. Chemistry 1996, 108, 359; A. Toepfer et al, Tetrahedron Letters 1995, 36, 9161; U. Sprengard et al, Angew. Chemistry 1995, 1_07, 1104; D. Zanini et al, Tetrahedron Letters 1995, 36, 7383; R. Roy et al, J. Chem. Commun. 1996, 201; W. Spevak et al, J. Med. Chem. 1996, 39, 1018; EAL Biessen et al, J. Med. Chem. 1995, 38, 1846; K. Yamada et al, Tetrahedron Letters 1995, 36, 9496; Lemieux et al, JACS 1975, 97, 4076; S. Koto et al, Bull. Chem. Soc. Jap. 1976, 49, 2639; PW Austin et al, J. Chem. Soz. C 1965, 1419; H. Paulsen, Angew. Chem. 1990, 1_02, 851; RR Schmidt et al, Angew. Chem. 1986, 98, 213; H. Kunz, Angew. Chem. 1987, 99, 297; J. Banoub, Chem. Review 1992, 92, 1167; H. Paulsen, Angew. Chem. 1982, 94, 184, RR Schmidt et al, Liebigs Ann. Chem. 1983, 1249; HA van Doren et al, Carbohydrate Research 1989, 194, 71, FD Trapper et al, J. Carbohydr. Chem. 1992, H, 751; T. Sugawara et al, Carbohydrate Research 1992, 230, 117, YC Lee et al, Biochemistry 1976, 5, 3956; Blomberg et al, J. Carbohydr. Chem. 1992, H, 751.

The pharmaceutical compositions according to the invention are likewise prepared in a manner known per se by suspending or dissolving the complex compounds according to the invention - if appropriate with addition of the additives customary in galenicals - in an aqueous medium and then optionally sterilizing the suspension or solution. Suitable additives are, for example, physiologically harmless buffers (such as tromethamine), additions of complexing agents or weak complexes (such as diethylenetriaminepentaacetic acid or the corresponding Ca-polysaccharide-polymer complexes) or - if necessary - electrolytes such as sodium chloride or - if necessary required - antioxidants such as ascorbic acid.

If suspensions or solutions of the agents according to the invention in water or physiological saline solution are desired for enteral administration or other purposes, they are mixed with one or more adjuvants common in galenics [e.g. Methyl cellulose, lactose, mannitol] and / the surfactant (s) [e.g. Lecithins, Tween®, Myrj®] and / or flavoring (s) for flavor correction [e.g. essential oils] mixed.

In principle, it is also possible to prepare the pharmaceutical compositions according to the invention without isolating the complex salts. In any case, special care must be taken to carry out the chelation so that the salts and salt solutions according to the invention are practically free of non-complexed toxic metal ions.

This can be ensured, for example, with the help of color indicators such as xylenol orange through control titrations during the manufacturing process become. The invention therefore also relates to processes for the preparation of the complex compounds and their salts. The final security is cleaning the isolated complex salt.

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

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

2. for radio diagnostics and radiotherapy in the form of their complexes with the radioisotopes of the elements with atomic numbers 27, 29, 31, 32,

37-39, 43, 49, 62, 64, 70, 75 and 77.

The agents according to the invention meet the diverse requirements for their suitability as contrast agents for magnetic resonance imaging. For example, after oral or parenteral application, they are excellently suited to improve the meaningfulness of the image obtained with the aid of an MRI scanner. They also show the high effectiveness that is necessary to burden the body with the smallest possible amount of foreign substances and the good tolerance that is necessary to maintain the non-invasive character of the examinations.

The good water solubility and low osmolality of the agents according to the invention make it possible to produce highly concentrated solutions, so that the volume of the circulatory system can be kept within reasonable limits and to compensate for the dilution with the body fluid, i.e. NMR diagnostics must be 100 to 1000 times more water soluble than for them NMR spectroscopy. Furthermore, the agents according to the invention not only have a high stability in vitro, but also a surprisingly high stability in vivo, so that the release or exchange of the ions which are not covalently bound in the complexes - in themselves toxic - within the time who the new Contrast agents are completely excreted again, takes place only extremely slowly.

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

Particularly low doses (below 1 mg / kg body weight) of organ-specific NMR diagnostics can be used, for example, to detect tumors and heart attacks.

Because of their favorable radioactive properties and the good stability of the complex compounds contained in them, the agents according to the invention are also suitable as radio diagnostic agents. Details of their application and dosage are e.g. in "Radiotracers for Medical Applications", CRC-Press, Boca Raton, Florida.

Another imaging method with radioisotopes is positron emission tomography, which is positron emitting isotopes such as ⁴³ Sc, ^ ⁴ Sc,

52p _θ] 55QO and 68Qa used (Heiss, WD; Phelps, ME; Positron Emission Tomography of Brain, Springer Verlag Berlin, Heidelberg, New York 1983).

The compounds according to the invention are surprisingly also suitable for differentiating 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 invention accumulate in malignant tumors (no diffusion into healthy tissues, but high permeability of tumor vessels), they can also support the radiation therapy of malignant tumors. This differs from the corresponding diagnostics only in the amount and type of isotope used. The aim is to destroy tumor cells by high-energy short-wave radiation with the shortest possible range. Interactions of the metals contained in the complexes (such as iron or gadolinium) with ionizing radiation (eg X-rays) or with neutron beams are used for this purpose. By This effect significantly increases the local radiation dose at the location where the metal complex is located (e.g. in tumors). In order to generate the same radiation dose in the malignant tissue, the radiation exposure for healthy tissue can be considerably reduced when using such metal complexes, thus avoiding adverse side effects for the patient. The metal complex conjugates according to the invention are therefore also suitable as a radio-sensitive substance in radiation therapy of malignant tumors (Mössbauer effects or neutron capture therapy can also be used). Suitable β-emitting ions are, for example, ⁴⁶ Sc, ⁴⁷ Sc, ⁴⁸ Sc, ^ ² Ga, ³ Ga _unc j 90γ. Suitable α-emitting ions with short half-lives are, for example, 211 Bi, ^{2 "} ! ² Bi, ² 1 ³ Bi and 14ßi, with ² 1 ² Bi being preferred. A suitable photon and electron emitting ion is ¹ ^ ³ Gd, which can be obtained from 157 (3 (3 by neutron capture.

When the therapeutic agents according to the invention are applied in vivo, they can be administered together with a suitable carrier such as serum or physiological saline and together with another protein such as human serum albumin. The dosage depends on the type of cellular disorder, the metal ion used and the type of imaging method.

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

Details of the use of radiotherapeutics are e.g. in R.W. Kozak et al. TIBTEC, October 1986, 262.

The agents according to the invention are outstandingly suitable as X-ray contrast agents, in particular for computed tomography (CT), it being particularly noteworthy that they do not show any signs of the anaphylaxis-like reactions known from the iodine-containing contrast agents in biochemical-pharmacological examinations. They are particularly valuable because of the favorable absorption properties in areas of higher tube voltages for digital subtraction techniques. In general, the agents according to the invention for use as X-ray contrast agents are dosed in amounts of 0.1-5 mmol / kg, preferably 0.25-1 mmol / kg, in analogy to meglumine diatrizoate.

Details of the use of X-ray contrast media are discussed, for example, in Barke, X-ray contrast media, G. Thieme, Leipzig (1970) and P. Thurn, E. Bücheier "Introduction to X-ray diagnostics", G. Thieme, Stuttgart, New York (1977).

Overall, it has been possible to synthesize new polymer-metal complexes that open up new possibilities in diagnostic and therapeutic medicine.

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

The abbreviations known to the person skilled in the art and known in the following examples, certain, frequently recurring compound names, are explained below by name and / or by structural formulas;

where = indicates the position in the molecules described below, via which the binding to the polymer P takes place.

1) DTPA: [bis- (2-aminoethyl) amine-N, N, N ', N ", N" - pentaacetic acid]

2) Gd-DTPA: Gadolinium complex of [bis- (2-aminoethyl) amine-N, N, N ', N ", N" - pentaacetic acid] monosodium salt

3) D

4) DTPA-tetra-tert-butyl ester (central carboxymethyl function):

3,9-bis (tert-butoxycarbonylmethyl) -6-carboxymethyl-3,6,9-triazaundecanoic acid di-tert-butyl ester

5) DTPA-tetra-tert-butyl ester (terminal carboxymethyl function):

6,9-bis (tert-butoxycarbonylmethyl) -3-carboxymethyl-3,6,9-triazaundecanoic acid 3,9-di-tert-butyl ester

6) Gly-Me-DOTA-tri-tert-butyl ester:

10- [1 - (Carboxymethylcarbamoyl) -ethyl] -1, 4.7, 10-tetraazacyclododecane-1, 4,7-tri-tert-butyl triacetic acid, sodium bromide:

7) Gd-Gly-e-DOTA:

Gadolinium complex of 10- [1 - (carboxymethylcarbamoyl) ethyl] -1, 4.7, 10- tetraaza-cyclododecane-1, 4,7-triacetic acid

8) 64 DSM polyamine:

9) 32mm DSM polyamine:

10) lactobionolactone:

O-ß-D-galactopyranosyl- (1 → 4) -D-glucono-1,5-lactone

11) Lactobionolactone Amide:

O-ß-D-galactopyranosyl- (1 → 4) -D-glucono-1,5-lactone amide

example 1

64-DSM polyamine (Gd-DTPA amide) 38 - [1- (4-thioureidophenyl) - ß-D-galactopyranosyl] 26-onjugate [as sodium salt]

a) Polyamide from 64-DSM polyamine and DTPA [as sodium salt]

5.0 g (0.7 mmol; corresponding to 44.8 mmol of free amine functions) 64-DSM polyamine are dissolved in 400 ml of distilled water and a pH of 9.5 is set with 1 molar sodium hydroxide solution. A total of 21.6 g (53.7 mmol) of DTPA monoanhydride monoethyl ester (preparation as described in EP 0263051 A1, EP 0450742 A1 and EP 0413405 A1) are then added in portions, the pH of the reaction solution being added of 1 molar sodium hydroxide solution is kept constant at 9.5. After the addition has ended, the mixture is stirred for a further 15 minutes at room temperature and then the pH of the reaction solution is adjusted to 11.5 by adding 32% sodium hydroxide solution. After a reaction time of 12 hours at room temperature, make up to a total volume of 800 ml with distilled water. The aqueous product solution thus obtained is ultrafiltered three times using a YM3 ultrafiltration membrane (Amicon) against distilled water. The remaining residue is made up to a volume of 500 ml with deionized water and the aqueous product solution is freeze-dried.

Yield: 14.04 g (88.3%; based on the polyamine used) as amorphous

powder

Water content: 8.46%

Elemental analysis (calculated on anhydrous substance): calc .: C 47.16 H 6.63 N 14.41 Na 7.69 found: C 46.88 H 6.79 N 14.32 Na 7.01

After elemental analysis and colorimetric titration with 0.0025 molar gadolinium sulfate standard solution in the presence of xylenol orange as an indicator, the degree of DTPA loading of the polymer is 59.9% (corresponding to 38 DTPA units per molecule). After quantitative ninhydrin reaction as well quantitative amino group determination using the TNBS method (see: Fields, R., J. Biochem. 1971, 124, 581-590 and further references: Chem. Rev., 60, 39 (1960); Tetrahedron, 34, ^ 2Q (1978); 47, 8791 (1991)], an average of 25.8 free amino functions per polymer molecule are present.

b) 64-DSM-polyamine (DTPA amide) 38 - [1 - (4-thioureidophenyl) - ß-D-galactopyranosyl] 26 conjugate [as sodium salt]

1.0 g (0.044 mmol; corresponding to 1.14 mmol of free amino functions) of the title compound from Example 1a) are dissolved in 80 ml of deionized water and the pH of the solution is adjusted to 9.5 with 1 molar hydrochloric acid. Then, in portions with a total of 1.55 g (4.9 mmol) of p-isothiocyanate-phenyl-β-D-galactopyranose (representation according to: M. Poπpipom et al., J. Med. Chem. 24, 1388; 1981 and J. Haeπsler et al., Bioconjugate Chem. 4, 85, 1993; For the general formation of isothiocyanates from the corresponding aniline precursors, see for example: DH Buss, IJ Goldstein; J. Chem. Soc., 1968, 1457-1461, McBroom CR, Samanen CH and Goldstein IJ, Methods Enzymol. 1972, 28, 212-219), the pH of the reaction solution being kept constant at 9.5 using 0.1 molar sodium hydroxide solution. After a reaction time of 12 hours at 22 ° C., the mixture is filtered and made up to a total volume of 250 ml with distilled water. Ultrafiltration of the reaction solution three times over a YM3 ultrafiltration membrane (Amicon) followed by freeze-drying of the remaining residue gives 1.30 g (95% of theory) of an amorphous and colorless solid. Water content: 3.8%

Elemental analysis (calculated on anhydrous substance): calc .: C 47.83 H 6.23 N 11, 79 S 2.70 Na 5.66 found: C 47.04 H 6.46 N 11, 53 S 2.77 Na 5.58

The ninhydrin reaction as well as the TNBS method, which are generally used for the quantitative determination of free amino functions, are negative, i. H. there are no free amino functions in the polymer. The percentage sulfur value of the elemental analysis of the titanium compound gives a degree of loading of the polymer with 40% β-D-galactose (corresponding to 26 galactose residues per molecule). The colorimetric phenolic

ERSÄΓZBLÄΓT (RULE 26) Determination of sulfuric acid (Dubois et al., Anal. Chem. 1956, 28, 3, 350) to determine the average pyranose loading level of the polyamine gives a mean statistical loading level of 39.68% of D-galactose residues per molecule and is thus to be regarded as identical to the degree of loading, which was determined solely by the percentage element ratio (found here: 26 D-galactose residues per molecule).

c) 64-DSM polyamine (Gd-DTPA amide) 38 - [1 - (4-thioureidophenyl) - ß-D-galactopyranosyl] 26-onjugate [as sodium salt]

1.27 g (0.0411 m mol; corresponding to 1.56 mmol of DTPA) of the title compound from Example 1b) are dissolved in 40 ml of sodium citrate buffer (pH 5.3) and at room temperature with a solution of 0.42 g (1 , 59 mmol) of gadolinium chloride in 10 ml of distilled water. After a reaction time of 15 minutes at room temperature, the reaction solution is adjusted to pH 7.2 with 1 molar sodium hydroxide solution. After three times of ultrafiltration through a YM3 ultrafiltration membrane (Amicon), the remaining residue is freeze-dried. Yield: 1.44 g (97.2% of theory) of the above-mentioned title compound as an amorphous and colorless solid. Water content: 3.89%

Elemental analysis (calculated on anhydrous substance): calc .: C 41.00 H 5.45 N 10.11 S 2.31 Gd 16.53Na 2.42 found: C 39.87 H 5.74 N 9.98 S 2, 28 Gd 16.58Na 2.42

The molar element ratio of gadolinium to sulfur thus results in an average degree of loading of the polymer of 60% Gd-DPTA and 40% D-galactose (corresponding to 38 Gd-DTPA units and 20-D-galactose residues per molecule). No free amino functions can be detected in the title compound by means of the quantitative ninhydrin reaction.

Example 2

64-DSM-polyamine- (Gd-DTPA) 38- [1- (4-thioureidophenyl) - -D-mannopyranosyl] 26 conjugate [as sodium salt] a) 64-DSM polyamine (DTPA amide) 38 - (1- (4-thioureidophenyl) - α-D-mannopyranosyl] 26 conjugate [as sodium salt]

In an analogous manner to that described for Example 1 b), the reaction of 0.75 g (0.033 mmol; corresponding to 0.855 mmol of free amine functions) of the title compound from Example 1 a) with 1.2 g (3.8 mmol) of p- lsothiocyaπatophenyl-α-D-mannopyranose [Presentation according to: Monsigny et al., Bio. Cell. 51, (1984), 187] after working up 0.93 g (92.4% of theory) of the above-mentioned title compound as an amorphous and colorless solid. Water content: 4.86%

Elemental analysis (calculated on anhydrous substance): calculated: C 47.83 H 6.23 N 11, 79 S 2.70 Na 5.66 found: C 47.76 H 6.51 N 11, 48 S 2.81 Na 5.74

The quantitative ninhydrin reaction of the title compound is negative, i. H. the polymeric product contains no free amino functions. The percentage sulfur of the elemental analysis of the title compound gives a degree of loading of the polymer with 40% -D-mannose (corresponding to 26 mannose residues per molecule).

b) 64-DSM-polyamine- (Gd-DTPA amide) 38- [1 - (4-thioureidophenyl) - -D- manπopyranosyl] 26 conjugate [as sodium salt]

0.85 g (0.027 mmol; corresponding to 1.04 mmol of DTPA) of the title compound from Example 2a) are described in analogy to that described for Example 1c), with 0.28 g (1.06 mmol) of gadolinium chloride used in the form of an aqueous Gadolinium chloride solution, complexed. After working up and freeze-drying, 958.2 mg (98.5% of theory) of the title compound are obtained as an amorphous powder. Water content: 6.14%

Elemental analysis (calculated on anhydrous substance): calc .: C 41.00 H 5.45 N 10.11 S 2.31 Gd 16.59Na 2.42 found: C 40.02 H 5.61 N 9.82 S 2.24 Gd 16.63 Na 2.46

ÖSÄΓZBLÄΓT (RULE 26) The molar elemental ratios of gadolinium to sulfur thus result in an average degree of loading of the polymer of 60.3% Gd-DPTA and 39.7% D-mannose (corresponding to 38 Gd-DTPA units and 26 D-mannose residues per molecule). No free amino functions can be detected in the title compound by means of the quantitative ninhydrin reaction. The T- | and T2 relaxivities of the title compound were determined in both water and plasma. The measurements were carried out at 40 ° C, a magnetic field strength of 0.47 Tesla, the T1 sequence: 180 ° - Tl - 90 ° [inversion recovery] and the T2 sequence: 90 ° - TE - 180 ° [CPMG].

All relaxivity determinations were carried out on a Minispec PC 20 and in all cases a solution of Gd-DTPA with a concentration of 1 mmol / L served as a reference sample.

Rl (water) = 13.9 ± 0.0 (L / mmol-s) R <l (plasma) = 17.6 ± 0.2 (L / mmol-s) R2 (water) = 16.8 ± 0.3 (L / mmol-s) R2 (plasma ) = 22.9 + 1.3 (L / mmol-s)

Example 3

64-DSM polyamine (Gd-DTPA amide) 38 - [1 - (4-thioureidophenyl) - ß-D-glucopyranosyl] 26 conjugate [as sodium salt]

a) 64-DSM polyamine (DTPA amide) 38 - [1 - (4-thioureidophenyl) - ß-D-glucopyranosyl] 26 conjugate [as sodium salt]

In an analogous manner to that described for Example 1b), the reaction of 0.9 g (0.04 mmol; corresponding to 1.4 mmol of free amine functions) of the title compound from Example 1 a) with 1.44 g (4.56 mmol) ) p-isothiocyanatophenyl-β-D-glucopyranose [Presentation according to: Monsigny et al., Bio. Cell. 51, (1984), 187; M. Ponpipom et al., J. Med. Chem. 24, 1388, (1981)] after working up 1, 12 g (90.6% of theory) of an amorphous and colorless solid. Water content: 3.75% Elemental analysis (calculated on anhydrous substance): calc .: C 47.83 H 6.23 N 11, 79 S 2.70 Na 5.66 found: C 47.76 H 6.51 N 11, 48 S 2.81 Na 5.74

The quantitative ninhydrin reaction of the title compound is negative, i. H. the polymeric product contains no free amino functions. The percentage sulfur value of the elemental analysis of the title compound gives a degree of loading of the polymer with 40% D-glucose (corresponding to 26 glucose residues per molecule).

b) 64-DSM-polyamine (Gd-DTPA amide) 38 - [1- (4-thioureidophenyl) - ß-D-glucopyranosyl] 26 conjugate [as sodium salt]

1.0 g (0.032 mmol; corresponding to 1.23 mmol of DTPA) of the title compound from Example 3a) are used in an analogous manner as described for Example 1 c) with 0.33 g (1.24 mmol) of gadolinium chloride, used in the form of an aqueous Gadolinium chloride solution, complexed. After working up and freeze-drying, 1.14 g (99.2% of theory) of the title compound is obtained as an amorphous powder. Water content: 5.47%

Elemental analysis (calculated on anhydrous substance): calc .: C 1.00 H 5.45 N 10.11 S 2.31 Gd 16.59Na 2.42 found: C 40.98 H 5.38 N 10.22 S 2, 40 Gd 16.43 Na 2.40

The molar elemental ratios of gadolinium to sulfur thus result in an average degree of loading of the polymer of 60% Gd-DPTA and 40% D-glucose (corresponding to 38 Gd-DTPA units and 26-D-glucose residues per molecule). No free amino functions can be detected in the title compound by means of the quantitative ninhydrin reactions. Example 4

64-DSM-polyamine- (Gd-DTPA amide) 38- [1- (4-thioureidophenyl) -α-L-fucopyranosyl] 26 conjugate [as sodium salt]

a) 64-DSM-polyamine (DTPA amide) 38- [1 - (4-thioureidophenyl) -α-L-fuco pyranosyl] 26 conjugate [as sodium salt]

In an analogous manner as described for Example 1 b), the implementation of

1.1 g (0.048 mmol; corresponding to 1.26 mmol of free amine functions) of the title compound from Example 1 a) with 1.65 g (5.57 mmol) of p-isothiocyanatophenyl-α-L-fucopyranose [representation according to: Tamio et al. , Jpn. Kokai Tokyo Koho, Jp 4211099 and Hilditch et al., J. Appl. Phycol. 3 (4), 345, (1991)] after working up 1.35 g (92.4% of theory) of the above-mentioned title compound as an amorphous and colorless solid. Water content: 8.03%

Elemental analysis (calculated on anhydrous substance): calc .: C 48.48 H 6.32 N 11, 95 S 2.74 Na 5.73 found: C 48.02 H 6.28 N 12.01 S 2.70 Na 5.81

The quantitative ninhydrin reaction of the title compound is negative, i. H. the polymeric product contains no free amino functions. The percentage sulfur value of the elemental analysis of the title compound gives a degree of loading of the polymer with 40% α-L-fucose (corresponding to 26 -L-fucose residues per molecule).

b) 64-DSM-polyamine (Gd-DTPA amide) 38- [1 - (4-thioureidophenyl) -α-L-fucopyranosyl] 26 conjugate [as sodium salt]

1.2 g (0.039 mmol; corresponding to 1.49 mmol of DTPA) of the title compound from Example 4a) are complexed with 0.40 g (1.51 mmol) of gadolinium chloride, used in the form of an aqueous gadolinium chloride solution, in analogy to that described for Example c) . After working up and freeze-drying, 1.37 g (98.5% of theory) of the title compound is obtained as an amorphous powder. Water content: 5.81% Elemental analysis (calculated on anhydrous substance) over C 41, 48 H 5.51 N 10.23 S 2.34 Gd 16.78 Na 2.45 found C 41, 31 H 5.54 N 10.17 S 2.38 Gd 16.55 Na 2.48

The molar elemental ratios of gadolinium to sulfur thus result in an average degree of loading of the polymer of 60% Gd-DPTA and 40% α-L-fucose (corresponding to 38 Gd-DTPA units and 26-α-L-fucose residues per molecule) In no quantitative amino functions can be detected in the title compound by means of the quantitative ninhydrin reaction

Example 5

64-DSM-Polyamιn (Gd-DTPA-Amιd) 38- [1- (4-Thιoureιdophenyl) -N-acetyl-ß-D-glucosamιπopyranosyl] 26-conjugate [as sodium salt]

a) 64-DSM-Polyamιn- (DTPA-Amιd) 38 - [1 - (4-Thιoureιdophenyl) -N-acetyl-ß-D-glucopyranosyl] 26- conjugate [as sodium salt]

In an analogous manner to that described for Example 1 b), the reaction of 0.9 g (0.04 mmol, corresponding to 1.26 mmol of free amine functions) of the title compound from Example 1a) with 1.61 g (4.56 mmol) ) 2- [acetylamine-2-deoxy] -1 - [4- (ιsothιocyanatophenyl)] -ß-D-glucopyranose [illustration Jonas, AJ et al, Biochem J (1990) 268, 1] after working up 926 mg (92, 2% d Th) of the abovementioned title compound as an amorphous and colorless solid, water content 7.68%

Elemental analysis (calculated on anhydrous substance) over C 44.10 H 4.62 N 9.26 S 3.32 Na 6.96 found C 44.21 H 6.58 N 9.31 S 3.28 Na 6.92

The quantitative ninhydrin reaction of the title compound is negative, ie the polymeric product does not contain any free amino functions. The percentage sulfur value of the elemental analysis of the title compound gives a result Degree of loading of the polymer with 40% 2- [acetylamino-2-deoxy] -1-phenyl-ß-glucopyranose (corresponding to 26 2- [acetylamino-2-deoxy] -1-phenyl-ß-D-glucopyranose residues per molecule).

b) 64-DSM polyamine (Gd-DTPA amide) 38 - [1 - (4-thioureidophenyl) -N-acetyl-ß-D-glucosaminopyranosyl] 26 ^_ conjugate [as sodium salt]

0.9 g (0.036 mmol; corresponding to 1.36 mmol of DTPA) of the title compound from Example 5a) are used in the same way as described for Example 1 c) with 0.38 g (1.45 mmol) of gadolinium chloride, used in the form of an aqueous Gadolinium chloride solution, complexed. After working up and freeze-drying, 1.26 g (92.3% of theory) of the title compound are obtained as an amorphous powder. Water content: 8.46%

Elemental analysis (calculated on anhydrous substance): calc .: C 41, 54 H 5.44 N 10.81 S 2.25 Gd 16.12 Na 2.36 found: C 41, 74 H 5.40 N 10.96 S 2.27 Gd 15.99 Na 2.28

The molar elemental ratios of gadolinium to sulfur thus result in an average degree of loading of the polymer of 60% Gd-DPTA and 40% N-acetyl-D-glucosamine (corresponding to 38 Gd-DTPA units and 26 N-acetyl-D-glucosamine residues per Molecule). No free amino functions can be detected in the title compound by means of the quantitative ninhydrin reaction.

Example 6

32-DSM polyamine (Gd-DTPA amide) s - [1 - (4-thioureidophenyl) -ß-D-galactopyranosyl] - _| 4-conjugate [as sodium salt]

a) Polyamide from 32-DSM polyamine and DTPA [as sodium salt]

A solution of 5.0 g (1.42 mmol, corresponding to 45.5 mmol of free

Amine functions) 32-DSM polyamine is dissolved in 400 ml of distilled water and adjusted to a pH of 9.5 with 1 molar sodium hydroxide solution. Subsequently a total of 21.9 g (54.6 mmol) of DTPA monoanhydride monoethyl ester is added in portions, the pH of the reaction solution being kept constant at 9.5 by adding 1 molar sodium hydroxide solution. After the addition has ended, the mixture is stirred for a further 15 minutes at room temperature and then the pH of the reaction solution is adjusted to 11.5 by adding 32% sodium hydroxide solution. After a reaction time of 12 hours at room temperature, make up to a total volume of 800 ml with distilled water. The aqueous product solution thus obtained was ultrafiltered three times using a YM3 ultrafiltration membrane (Amicon) against distilled water. The remaining residue is made up to a volume of 500 ml with deionized water and the aqueous product solution is freeze-dried.

Yield: 15.5 g (91.2% of theory; based on the polyamine used) as an amorphous powder. Water content: 7.43%

Elemental analysis (calculated on anhydrous substance): calc .: C 43.74 H 7.21 N 13.57 Na 13.83 found: C 43.21 H 7.18 N 13.82 Na 13.94

After elemental analysis and colorimetric titration with 0.0025 molar gadolinium sulfate standard solution in the presence of xylenol orange as an indicator, the degree of DTPA loading of the polymer is 55.8% (corresponding to 18 DTPA units per molecule). After quantitative ninhydrin reaction and quantitative amino group determination using the TNBS method (see: Fields, R., J. Biochem. 1971, 124, 581-590 and further references: Chem. Rev., 60, 39 (1960) ; Tetrahedron, 34, 1285 (1978); 47, 8791 (1991)] there are an average of 14 free amino functions per polymer molecule.

b) 32-DSM-polyamine (DTPA amide) ι Q - [1 - (4-thioureidophenyl) - ß-D-galactopyranosyl] i4 conjugate [as sodium salt]

In an analogous manner to that described for Example 1b), the reaction of 1.0 g (0.0835 mmol; corresponding to 1.17 mmol of free amine functions) of the title compound from Example 6a) with 1.60 g (5.03 mmol) ) p-isothiocyanato- phenyl-β-D-galactopyranose after working up 1.25 g (91.3% of theory) of the above-mentioned title compound as an amorphous and colorless solid. Water content: 8.36%

Elemental analysis (calculated on anhydrous substance): calc .: C 45.34 H 6.65 N 11, 12 S 2.74 Na 10, 11 found: C 44.98 H 6.51 N 11, 02 S 2.68 Na 10.14

The quantitative ninhydrin reaction of the title compound is negative, i. H. the polymeric product contains no free amino functions. The percentage of sulfur in the elemental analysis of the title compound gives a degree of loading of the polymer with 44% β-D-galactose (corresponding to 14 β-galactose residues per molecule). The colorimetric determination of phenol-sulfuric acid (Dubois et al., Anal. Chem. 1956, 28, 3, 350) for determining the average pyranose loading level of the polyamine provides a mean statistical loading level of 43.75% of β-D -Galactose residues per molecule and is therefore to be regarded as identical to the degree of loading, which was determined solely by the percentage element ratios (found here: 26 β-D-galactose residues per molecule).

c) 32-DSM-polyamine- (Gd-DTPA-amide) ι β- [1 - (4-thioureidophenyl) -ß-D-galactopyranosyl] - | 4-conjugate [as sodium salt]

1.0 g (0.061 mmol; corresponding to 1.10 mmol of DTPA) of the title compound from Example 6b) are described in analogy to Example 1c) with 0.29 g (1.15 mmol) of gadolinium chloride used in the form of an aqueous gadolinium chloride solution , complexed. After working up and freeze-drying, 1.05 g (96.2% of theory) of the title compound is obtained as an amorphous powder. Water content: 4.93%

Elemental analysis (calculated on anhydrous substance): calc .: C 41, 33 H 6.06 N 10.14 S 2.50 Gd 15.76 Na 2.30 found: C 41, 28 H 5.97 N 10.08 S 2.52 Gd 15.80 Na 2.28

The molar element ratio of gadolinium to sulfur thus results in an average degree of loading of the polymer of 56% Gd-DPTA and 44% ß-D-galactose (corresponding to 18 Gd-DTPA units and 14 D-galactose residues per molecule). No free amino functions can be detected in the title compound by means of the quantitative ninhydrin reaction.

Example 7

32-DSM-polyamine- (Gd-DTPA) i 8- [1 - (4-thioureidopheπyl) -α-D-mannopyranosyl] i 4-conjugate [as sodium salt]

a) 32-DSM polyamino (DTPA amide) -ι s - [1 - (4-thioureidophenyl) -α-D-mannopyraπosyl] - | 4-conjugate [as sodium salt]

In an analogous manner to that described for Example 1), the reaction of 0.95 g (0.079 mmol; corresponding to 1.11 mmol of free amine functions) of the title compound from Example 6a) with 1.52 g (4.77 mmol) p - Isothiocyanatophenyl-ß-D-maππopyranose after working up 1.22 g (96.2% of theory) of the above-mentioned title compound as an amorphous and colorless solid.

Water content: 5.93%

Elemental analysis (calculated on anhydrous substance): calc .: C 45.34 H 6.65 N 11, 12 S 2.74 Na 10.1 1 found: C 45.20 H 6.49 N 1 1, 20 S 2 , 66 Na 10.21

The quantitative ninhydrin reaction of the title compound is negative, i. H. the polymeric product contains no free amino functions. The percentage sulfur value of the elemental analysis of the title compound gives a degree of loading of the polymer with 44% αD-mannose (corresponding to 14 α-D-maππose residues per molecule).

b) 32-DSM-Poiyamiπ- (Gd-DTPA amide) -ι s- [1 - (4-thioureidophenyl) -α -D- mannopyraπosylji 4 conjugate [as sodium salt]

REPLACEMENT SHEET (RULE 21 1.1 g (0.067 mmol; corresponding to 1.21 mmol of DTPA) of the title compound from Example 7a) are used in the same way as described for Example 1 c) with 0.30 g (1.22 mmol) of gadolinium chloride, used in the form of an aqueous Gadolinium chloride solution, complexed. After working up and freeze-drying, 1.18 g (98.6% of theory) of the title compound are obtained as an amorphous powder. Water content: 3.98%

Elemental analysis (calculated on anhydrous substance): calc .: C 41, 33 H 6.06 N 10.14 S 2.50 Gd 15.76 Na 2.30 found: C 41, 27 H 6.10 N 10.21 S 2 , 54 Gd 15.83 Na 2.36

The molar elemental ratios of gadolinium to sulfur thus result in an average degree of loading of the polymer of 56% Gd-DPTA and 44% α-D-mannose (corresponding to 18 Gd-DTPA units and 14-D-mannose residues per molecule). No free amino functions can be detected in the title compound by means of the quantitative ninhydrin reaction.

Example 8

32-DSM polyamine (Gd-DTPA amide) i 8 - [1 - (4-thioureidophenyl) - ß-D-glucopyranosyl] -] 4-conjugate [as sodium salt]

a) 32-DSM-polyamine- (DTPA amide) ι 8 - [1 - (4-thioureidophenyl) - ß-D-glucopyranosyl] i4 conjugate [as sodium salt]

In an analogous manner to that described for Example 1 b), the reaction gives 1.2 g (0.10 mmol; corresponding to 1.4 mmol of free amine functions)

Title compound from Example 6a) with 1.92 g (6.03 mmol) p-

Isothiocyanatophenyl-β-D-glucopyranose after working up 1.50 g (91.6% of theory

Th.) Of the above title compound as amorphous and colorless

Solid. Water content: 6.49%

Elemental analysis (calculated on anhydrous substance): calc .: C 45.34 H 6.65 N 11, 12 S 2.74 Na 10.11 found: C 44.98 H 6.60 N 11, 19 S 2.70 Na 10.19

The quantitative ninhydrin reaction of the title compound is negative, i. H. the polymeric product contains no free amino functions. The percentage of sulfur in the elemental analysis of the title compound gives a degree of loading of the polymer with 44% β-D-glucose (corresponding to 14 β-D-glucose residues per molecule).

b) 32-DSM-polyamine- (Gd-DTPA-amide) ι Q - [1 - (4-thioureidophenyl) - ß-D-glucopyranosyl] i4 conjugate [as sodium salt]

1.40 g (0.085 mmol; corresponding to 1.54 mmol of DTPA) of the title compound from Example 8a) are complexed with 0.38 g (1.56 mmol) of an aqueous gadolinium chloride solution in analogy to that described for Example 1 c). After working up and freeze-drying, 1.50 g (98.4% of theory) of the title compound are obtained as an amorphous powder. Water content: 7.33%

Elemental analysis (calculated on anhydrous substance): calc .: C 41, 33 H 6.06 N 10.14 S 2.50 Gd 15.76 Na 2.30 found: C 41, 22 H 6.10 N 10.22 S 2 , 48 Gd 15.71 Na 2.28

The molar elemental ratios of gadolinium to sulfur thus result in an average degree of loading of the polymer of 56% Gd-DPTA and 44% β-D-glucose (corresponding to 18 Gd-DTPA units and 14 β-D-glucose residues per molecule). No free amino functions can be detected in the title compound by means of the quantitative ninhydrin reaction.

Example 9

32-DSM-Polyamiπ- (Gd-DTPA amide) -i 8- [1- (4-thioureidophenyl) -α-L-fucopyranosyl] - | 4-conjugate [as sodium salt] a) 32-DSM-polyamine (DTPA amide) ι 8- [1 - (4-thioureidophenyl) -α-L-fucopyranosyl] i4 conjugate [as sodium salt]

In an analogous manner to that described for example 1b), the reaction gives 1.0 g (0.083 mmol; corresponding to 1.17 mmol of free amine functions)

Title compound from Example 6a) with 1.50 g (5.03 mmol) of p-

Isothiocyanatophenyl-α-L-fucopyranose after working up 1.25 g (93.6% of theory

Th.) Of the above title compound as amorphous and colorless

Solid. Water content: 6.74%

Elemental analysis (calculated on anhydrous substance): calc .: C 45.97 H 6.74 N 11, 28 S 2.78 Na 10.25 found: C 46.03 H 6.81 N 11, 32 S 2.75 Na 10.19

The quantitative ninhydrin reaction of the title compound is negative, i. H. the polymeric product contains no free amino functions. The percentage sulfur value of the elemental analysis of the title compound gives a degree of loading of the polymer with 44% α-L-fucose (corresponding to 14 α-L-fucose residues per molecule).

b) 32-DSM-polyamine (DTPA amide) 8- [- (4-thioureidophenyl) -α-L-fucopyranosyl] -] 4-conjugate [as sodium salt]

1.1 g (0.068 mmol; corresponding to 1.22 mmol of DTPA) of the title compound from Example 9a) are used in the same way as described for Example 1c) with 0.31 g (1.24 mmol) of gadolinium chloride, used in the form of an aqueous gadolinium chloride solution , complexed. After working up and freeze-drying, 1.19 g (98.6% of theory) of the title compound is obtained as an amorphous powder. Water content: 7.23%

Elemental analysis (calculated on anhydrous substance): calc .: C 41, 85 H 6.14 N 10.27 S 2.53 Gd 15.96 Na 2.33 found: C 41, 81 H 6.08 N 10.21 S 2.50 Gd 16.00 Na 2.34 The molar elemental ratios of gadolinium to sulfur thus result in an average degree of loading of the polymer of 56% Gd-DPTA and 44% α-L-fucose (corresponding to 18 Gd-DTPA units and 14 α-L-fucose residues per molecule). No free amino functions can be detected in the title compound by means of the quantitative ninhydrin reaction.

Example 10

32-DSM polyamine (Gd-DTPA amide) ι g- [1 - (4-Thioureidoρheπyl) -N-acetyl-ß-D-glucosaminopyranosyl] i4 conjugate [as sodium salt]

a) 32-DSM-polyamine (DTPA amide) ι g- [1 - (4-thioureidophenyl) -N-acetyl- ß-D-glucopyranosyl] i4 conjugate [as sodium salt]

In an analogous manner to that described for Example 1 b), the reaction of 1.4 g (0.11 mmole; corresponding to 1.63 mmol of free amine functions) of the title compound from Example 6a) with 2.50 g (7.04 mmol ) 2- [Acetylamino-2-deoxy] -1 - [4- (isothiocyanatophenyl)] -ß-D-glucosaminopyranose [Illustration: Jonas, AJ et. al., Biochem. J. (1990) 268, 1] after working up 1.25 g (93.6% of theory) of the above-mentioned title compound as an amorphous and colorless solid. Water content: 7.33%

Elemental analysis (calculated on anhydrous substance): calc .: C 44.11 H 6.43 N 15.13 S 2.55 Na 9.41 found: C 44.08 H 6.22 N 15.11 S 2.58 Na 9.38

The quantitative ninhydrin reaction of the title compound is negative, i. H. the polymeric product contains no free amino functions. From the percentage sulfur value of the elemental analysis of the title compound, a degree of loading of the polymer with 44% N-acetyl-D-glucosamine (corresponding to 14 N-acetyl-β-D-glucosamine residues per molecule) results.

b) 32-DSM-polyamine (Gd-DTPA amide) ₁ 3 - [1 - (4-thioureidophenyl) -N-acetyl-ß-D-giucosaminopyranosyl] <i 4 conjugate [as sodium salt]

S - SÄΓZBLAΓT (RULE 26) 1.6 g (0.09 mmol; corresponding to 1.64 mmol of DTPA) of the title compound from Example 10a) are used in the same way as described for Example 1 c) with 0.42 g (1.66 mmol) of gadolinium chloride used in the form an aqueous gadolinium chloride solution, complexed. After working up and freeze-drying, 1.70 g (98.7% of theory) of the title compound are obtained as an amorphous powder. Water content: 5.51%

Elemental analysis (calculated on anhydrous substance): calc .: C 40.45 H 5.90 N 13.88 S 2.34 Gd 14.76 Na 2.16 found: C 40.36 H 5.87 N 13.94 S 2.34 Gd 14.81 Na 2.20

The molar elemental ratios of gadolinium to sulfur thus result in an average degree of loading of the polymer of 56% Gd-DPTA and 44% N-acetyl-ß-D-glucosamine (corresponding to 18 Gd-DTPA units and 14-N-acetyl-ß -D-glucosamine residues per molecule). No free amino functions can be detected in the title compound by means of the quantitative ninhydrin reaction.

Example 11

64-DSM-polyamine (Gd-Gly-Me-DOTA amide) 28 - (α-D-mannopyranosyl-1-thio-3-propionic acid amide) 3ß conjugate

a) N- (2-Bromopropionyl) glycine benzyl ester

60.97 g (355.7 mmol) α are slowly added dropwise at 0 ° C. to 100 g (296.4 mmol) of glycine benzyl ester of p-toluenesulfonic acid salt and 89.98 g (889.2 mmol) of triethylamine, dissolved in 500 ml of methylene chloride -Bromopropionyl chloride too. The temperature is kept between 0 ° C - 5 ° C. 1000 ml of 5% aqu. Add hydrochloric acid and separate the organic phase. The organic phase is again with 500 ml of 5% aqu. Washed hydrochloric acid, dried over magnesium sulfate and evaporated to dryness in vacuo. The residue is recrystallized from di-isopropyl ether. Yield: 69.39 g (78% of theory) Egg entaranalysis: calc .: C 48.02 H 4.70 N 4.67 Br 26.62 found: C 47.91 H 4.82 IM 4.51 Br 26.47

b) 1 - [1-Benzyloxycarbonylmethylcarbamoyl) -ethyl] -1, 4.7, 10-tetraazacyclododecane

To 86.14 g (500 mmol) 1, 4,7,10-tetraazacyclododecane, dissolved in 1000 ml

Chloroform, 50 g (166.6 mmol) of the title compound from Example 11a) are added and the mixture is stirred at room temperature for 24 hours. It is washed 3 times with 600 ml of water, the organic phase is dried over magnesium sulfate and evaporated to dryness in vacuo. Yield: 53.48 g (82% of theory) of a light yellow oil. Water content: 1.5%

Elemental analysis (calculated on anhydrous substance): calc .: C 61, 36 H 8.50 N 17.89 found: C 62.03 H 8.75 N 17.36

c) 10- [1-Benzyloxycarbonylmethylcarbamoyl) -ethyl] -1, 4.7, 10-tetraazacyclododecane-1, 4,7-tri-tert-butyl ester, sodium bromide

To 50 g (127.7 mmol) of the title compound from Example 11b) and 54.14 g (510.8 mmol) of sodium carbonate in 500 ml of acetonitrile are added dropwise 82.20 g (421.4 mmol) of bromoacetic acid tert at room temperature. -butyl ester and stirred at 60 ° C for 12 hours. The mixture is cooled to 0 ° C. and the salts are filtered off. The filtrate is evaporated to dryness and the residue is chromatographed on silica gel (mobile solvent: methylene chloride / methanol = 20: 1). Yield: 90.83 g (85% of theory) of a colorless solid

Elemental analysis: calc .: C 54.54 H 7.59 N 8.37 Na 2.75 Br 9.55 found: C 54.37 H 7.71 N 8.21 Na 2.83 Br 9.69 d) 10- [1-carboxymethylcarbamoyl) ethyl] -1, 4.7, 10-tetraazacyclododecane-1, 4,7-tri-tert-butyl ester, sodium bromide

90 g (107.5 mmol) of the title compound from Example 11 c) are dissolved in 1000 ml of isopropanol and 5 g of palladium catalyst (10% Pd / C) are added. One hydrates overnight at room temperature. The catalyst is filtered off and the filtrate is evaporated to dryness. The residue is recrystallized from dioxane. Yield: 77.06 g (96% of theory) of a crystalline solid

Elemental analysis: calc .: C 49.86 H 7.69 N 9.38 Na 3.08 Br 10.70 found: C 49.73 H 7.79 N 9.21 Na 3.19 Br 10.83

e) 64-DSM polyamine (Gly-Me-DOTA-tri-t.-butyl ester amide) 28-onjugate

20.0 g (26.8 mmol) of Gly-Me-DOTA-tri-tert-butyl ester (title compound of Examples 11d) are dissolved in 100 ml of absolute dimethylformamide at room temperature and with stirring, 3.7 g (32.16 mmol ) N-

Hydroxysuccinimide added. The clear reaction mixture is then cooled to 0 ° C. and a total of 6.6 g (32.16 mmol) of dicyclohexylcarbodiimide is added at this temperature in portions. After a reaction time of 30 minutes at 0 ° C and 2.5 hours at room temperature, the precipitated dicyclohexylurea is filtered off. The filtrate thus obtained is then slowly added dropwise at room temperature to a stirred solution of 5.0 g (0.7 mmol; corresponding to 44.8 mmol of free amino functions) 64-DSM polyamine and 4.5 g (44.8 mmol) Triethylamine in 40 ml of absolute DMF. After a reaction time of 12 hours at room temperature, the mixture is evaporated to dryness in vacuo and 200 ml of dichloromethane are added to the remaining residue. After filtering off insoluble constituents, the organic phase is washed with saturated sodium bicarbonate solution and twice with water. After drying the organic product phase over sodium sulfate, the mixture is filtered and the solvent is stripped off in vacuo. 13.03 g (82.4% of theory; based on the polyamine used) of a slightly yellow-colored oil are obtained, which is immediately used for the subsequent synthesis steps without further purification. After quantitative Ninhydrin reaction of the above-mentioned title compound has an average of 35.8 free amine functions per molecule.

Elemental analysis: calc .: C 65.29 H 10.65 N 16.12 found: C 64.95 H 10.58 N 16.22

f) 64-DSM-polyamine- (Gly-Me-DOTA-tri-tert-butyl ester amide) 28- (3- (2,3,4,6-tetra-O-acetyl) -1 -thio-α- D-mannopyranosyl-3-propione-amide) 36 conjugate

587 mg (5.8 mmol) of triethylamine are added to a stirred solution of 3.0 g (0.132 mmol; corresponding to 4.8 mmol of free amine functions) of the title compound from Example 12a), dissolved in 100 ml of absolute dichloromethane. Then with a solution of 2.8 g (5.28 mmol) of 3- (2,3,4,6-tetra-O-acetyl-1-thio-α-D-mannopyranosyl) propionic acid N-hydroxysuccinimide ester ( Representation according to: J. Haensler et al., Bioconjugate Chem. 4, 85, (1993); Chipowsky, S., and Lee.YC (1973), Synthesis of 1-thio-aldosides; Carbohydrate Research 31, 339-346) added dropwise in 60 ml of dichloromethane. After a reaction time of 12 hours at

The room temperature is made up to a total volume of 500 ml with dichloromethane. The organic phase is washed successively twice with water and twice with saturated sodium bicarbonate solution and dried over sodium sulfate. After filtering, the solvent is removed to dryness in vacuo. 4.60 g (92.6% of theory) of the title compound are obtained as a colorless oil. The quantitative ninhydrin reaction of the title compound is negative, i. H. free amino functions are no longer detectable in the polymer.

Elemental analysis: calc .: C 58.69 H 8.51 N 8.51 S 3.07 found: C 58.73 H 8.60 N 8.60 S 3.10

The percentage sulfur value of the elemental analysis of the title compound gives an average degree of loading of the polymer with 56.7% α-D-mannose tetraacetate (corresponding to 36 α-D-mannose residues per molecule). g) 64-DSM-polyamine (Gly-Me-DOTA amide) 28 - (- α-D-mannopyranosyl-1 - thio-3-propionamide) 36 conjugate

4.0 g (0.10 mmol) of the title compound from Example 12b) are dissolved in 100 ml of absolute dichloromethane. A total of 75 ml of trifluoroacetic acid is then added dropwise at 0 ° C. After a reaction time of 12 hours at room temperature, the mixture is evaporated to dryness in vacuo. The remaining residue is mixed with 100 ml of water and stripped again to dryness in vacuo. The residue is dissolved in 100 ml of methanol and 50 ml of a 32% aqueous ammonia solution are added at room temperature. After a reaction time of 12 hours at room temperature, the solvent is removed in vacuo and the remaining residue is dissolved in 200 ml of distilled water. The aqueous product solution is extracted twice with 60 ml of diethyl ether each time. The aqueous product solution is obtained by adding 10% aqu. Hydrochloric acid brought to pH 3.0, made up to a total volume of 800 ml with water and ultrafiltered three times using a YM3 ultrafiltration membrane (Amicon) against distilled water. The remaining residue is made up to a volume of 500 ml with deionized water and freeze-dried. Yield: 2.2 g (76.8% of theory) as an amorphous powder Water content: 7.37%

Elemental analysis (calculated on anhydrous substance): calc .: C 50.68 H 7.76 N 13.12 S 4.01 found: C 50.42 H 7.91 N 13.30 S 4.11

h) 64-DSM-polyamine- (Gd-Gly-Me-DOTA-amide) 28 - (α-D-mannopyranosyl-1-thio-3-propionic acid amide) 3g conjugate

2.0 g (0.07 mmol; corresponding to 1.94 mmol of Gly-Me-DOTA) of the title compound from Example 12c) are dissolved in 50 ml of distilled water and the pH of the solution is adjusted by dropwise addition with 10% aqueous hydrochloric acid adjusted to 2.0. Then 0.52 g (1.98 mmol) of gadolinium chloride is added at room temperature and the mixture is stirred Reaction solution for 12 hours at 60 ° C. At room temperature, the reaction solution is brought to pH 7.2 by adding 1 molar sodium hydroxide solution and made up to a total volume of 800 ml with distilled water. After three times ultrafiltration against distilled water over a YM3 ultrafiltration membrane (Amicon), the remaining residue is freeze-dried. 2.27 g (98.2% of theory; based on the polyamine used) of the title compound are obtained as an amorphous, colorless powder with a water content of 6.39%.

Elemental analysis (calculated on anhydrous substance): calc .: C 44.08 H 6.50 N 11, 41 S 3.48 Gd 13.29 found: C 44.15 H 6.61 N 11, 37 S 3.54 Gd 13.35

The molar element ratio of sulfur to gadolinium (1, 30) results in an average degree of loading of the polymer of 43.7% Gd-Gly-Me-DOTA and 56.3% α-D-mannose (corresponding to 28 Gd-Gly-Me -DOTA units and 36 α-D-mannose residues per molecule). No free amino functions can be detected in the title compound by means of a quantitative ninhydrin reaction.

Example 12

64-DSM-polyamine (Gd-Gly-Me-DOTA amide) 28- (α-L-fucopyranosyl-1-thio-3-propionic acid amide) 35 conjugate

a) 64-DSM-polyamine- (Gly-Me-DOTA-tri-tert-butyl ester amide) 28- [(3- (2.3 _I 4 ₎ 6-tri-O-acetyl) -1 -thio- α -L-fucopyranosyl-3-propion-amide) 36 conjugate

A stirred solution of 2.5 g (0.11 mmol; corresponding to 3.96 mmol of free amine functions) of the title compound from Example 11 e) in 80 ml of absolute dichloromethane is mixed with 486 mg (4.8 mmol) of triethylamine at room temperature _. transferred. Then with a solution of 2.0 g (4.35 mmol) of 3- (2,3,4,6-tri-O-acetyl-1-thio-α-L-fucopyranosyl) propionic acid N-hydroxysuccinimide ester ( Representation in analogy to: J. Haensler et al., Bioconjugate Chem. 4, 85, (1993); Chipowsky, S., and Lee.YC (1973), Synthesis of 1 -thio-aidosides; Carbohydrate Research 31, 339- 346) dropwise in 60 ml dichloromethane

SPARE BLADE (RULE 26) transferred. After a reaction time of 12 hours at room temperature, the mixture is made up to a total volume of 500 ml with dichloromethane. The organic phase is washed successively twice with water and twice with saturated sodium bicarbonate solution and dried over sodium sulfate. After filtering, the solvent is removed to dryness in vacuo. 3.32 g (85.1% of theory; based on the polyamine used) of the title compound are obtained as a colorless oil. The quantitative ninhydrin reaction of the title compound is negative, ie no free amino functions can be detected in the polymer.

Elemental analysis: calc .: C 59.71 H 8.81 N 10.24 S 3.23 found: C 59.09 H 8.60 N 9.98 S 3.17

b) 64-DSM-polyamine (Gly-Me-DOTA amide) 28 ~ (α-L-fucopyraπosyl-1-thio-3-propionic acid amide) 36 conjugate

4.0 g (0.112 mmol) of the title compound from Example 12a) are dissolved in 80 ml of absolute dichloromethane. A total of 60 ml of trifluoroacetic acid is then added dropwise at 0 ° C. After a reaction time of 12 hours at room temperature, the mixture is evaporated to dryness in vacuo. The remaining residue is mixed with 80 ml of water and again stripped to dryness in vacuo. The residue is dissolved in 100 ml of methanol and 50 ml of a 32% aqueous ammonia solution are added at room temperature. After a reaction time of 12 hours at room temperature, the solvent is removed in vacuo and the remaining residue is dissolved in 200 ml of distilled water. The aqueous product solution is extracted twice with 60 ml of diethyl ether each time. The aqueous product solution is mixed with 10% aqu. Hydrochloric acid brought to pH 3.0, made up to a total volume of 800 ml with water and ultrafiltered three times using a YM3 ultrafiltration membrane (Amicon) against distilled water. The remaining residue is made up to a volume of 500 ml with deionized water and freeze-dried. Yield: 2.37 g (76.2% of theory) of the above-mentioned title compound colorless and amorphous powder. Water content: 7.43% Elemental analysis (calculated on anhydrous substance): calc .: C 52.79 H 8.09 N 13.67 S 4.17 found: C 52.66 H 7.99 N 13.40 S 4.14

c) 64-DSM-polyamine (Gd-Gly-Me-DOTA amide) 28- (α-L-fucopyranosyl-1 - thio-3-propionic acid amide) 3g conjugate

2.0 g (0.072 mmol; corresponding to 2.02 mmol of Gly-Me-DOTA) of the title compound from Example 12b) are dissolved in 40 ml of distilled water and the pH of the solution is brought to 2.0 by dropwise addition with 10% aqueous hydrochloric acid set. Then 534 mg (2.04 mmol) of gadolinium chloride are added at room temperature and the reaction solution is stirred at 60 ° C. for 12 hours. At room temperature, the reaction solution is brought to pH 7.2 by adding 1 molar sodium hydroxide solution and made up to a total volume of 500 ml with distilled water. After three times ultrafiltration against distilled water over a YM3 ultrafiltration membrane (Amicon), the remaining residue is freeze-dried. 2.26 g (98.1% of theory; based on the polyamine used) of the title compound are obtained as an amorphous, colorless powder with a water content of 9.09%.

Elemental analysis (calculated on anhydrous substance): calc .: C 45.57 H 6.72 N 11, 80 S 3.80 Gd 13.74 found: C 45.68 H 6.86 N 11.08 S 3.59 Gd 13.56

The molar element ratio of sulfur to gadolinium (1, 32) results in an average degree of loading of the polymer of 43.8% Gd-Gly-Me-DOTA and 56.2% α-L-fucose (corresponding to 28 Gd-Gly-Me -DOTA units and 36 α-L-fucose residues per molecule). No free amino functions can be detected in the title compound by means of a quantitative ninhydrin reaction. Example 13

64-DSM-polyamine (Gd-Gly-Me-DOTA amide) 28- (ß-D-glucopyranosyl-1-thio-3-propionic acid amide) 3g conjugate

a) 64-DSM-polyamine- (Gly-Me-DOTA-tri-tert-butyl ester amide) 28- [(3- (2,3,4,6-tetra-O-acetyl) -1 -thio- ß -D-glucopyranosyl-3-propione-amide] 36 conjugate

A stirred solution of 2.5 g (0.11 mmol; corresponding to 3.96 mmol of free amine functions) of the title compound from Example 11e) in 80 ml of absolute dichloromethane is mixed with 486 mg (4.8 mmol) of triethylamine at room temperature. Then with a solution of 2.31 g (4.35 mmol) of 3- (2, 3,4,6-tetra-O-acetyl-1-thio- ß-D-glucopyranosyl) -propionic acid-N-hydroxysuccinimide- ester (representation in analogy to: J. Haensler et al., Bioconjugate Chem. 4, 85, (1993); Chipowsky, S., and Lee.YC (1973), Synthesis of 1 -thio-aldosides; Carbohydrate Research 31, 339-346) added dropwise in 60 ml of dichloromethane. After a reaction time of 12 hours at room temperature, the mixture is made up to a total volume of 500 ml with dichloromethane. The organic phase is washed successively twice with water and twice with saturated sodium bicarbonate solution and dried over sodium sulfate. After filtering, the solvent is removed to dryness in vacuo. 3.70 g (89.4% of theory) of the title compound are obtained as a colorless oil. The quantitative ninhydrin reaction of the title compound is negative, i. H. free amino functions are no longer detectable in the polymer.

Elemental analysis: calc .: C 58.69 H 8.51 N 9.67 S 3.07 found: C 58.60 H 8.42 N 9.88 S 3.14

b) 64-DSM-polyamine- (Gly-Me-DOTA-amide) 28- (ß-D-glucopyranosyl-1-thio-3-propionic acid amide) 36 conjugate

3.5 g (0.087 mmol) of the title compound from Example 13a) are dissolved in 80 ml of absolute dichloromethane. A total of 60 ml of trifluoroacetic acid is then added dropwise at 0 ° C. After a reaction time of 12 hours at room temperature, the mixture is evaporated to dryness in vacuo. The the remaining residue is mixed with 80 ml of water and again stripped to dryness in vacuo. The residue is dissolved in 100 ml of methanol and 50 ml of a 32% strength aqueous ammonia solution are added at room temperature. After a reaction time of 12 hours at room temperature, the solvent is removed in vacuo and the remaining residue is dissolved in 200 ml of distilled water. The aqueous product solution is extracted twice with 60 ml of diethyl ether each time. The aqueous product solution is mixed with 10% aqu. Hydrochloric acid brought to pH 3.0, made up to a total volume of 800 ml with water and ultrafiltered three times using a YM3 ultrafiltration membrane (Amicon) against distilled water. The remaining residue is made up to a volume of 500 ml with deionized water and freeze-dried. Yield: 1.97 g (78.8% of theory) as an amorphous powder Water content: 5.45%

Elemental analysis (calculated on anhydrous substance): calc .: C 50.68 H 7.76 N 13.12 S 4.01 found: C 50.48 H 7.89 N 13.42 S 4.14

c) 64-DSM-polyamine (Gd-Gly-Me-DOTA amide) 28- (ß -D-glucopyranosyl-1 - thio-3-propionic acid amide) 35 conjugate

1.5 g (0.052 mmol; corresponding to 1.47 mmol of Gly-Me-DOTA) of

The title compound from Example 13b) are dissolved in 40 ml of distilled water and the pH of the solution is adjusted to 2.0 by dropwise addition with 10% aqueous hydrochloric acid. Then 393 mg (1.50 mmol) of gadolinium chloride are added at room temperature and the reaction solution is stirred at 60 ° C. for 12 hours. At room temperature, the reaction solution is brought to pH 7.2 by adding 1 molar sodium hydroxide solution and made up to a total volume of 500 ml with distilled water. After three times ultrafiltration against distilled water over a YM3 ultrafiltration membrane (Amicon), the remaining residue is freeze-dried. 1.66 g (96.3% of theory) of the title compound are obtained as an amorphous, colorless powder with a water content of 8.47%. Elemental analysis (calculated on anhydrous substance): calc .: C 44.08 H 6.50 N 11, 41 S 3.48 Gd 13.29 found: C 44.18 H 6.59 N 1 1, 52 S 3, 55 Gd 13.12

The molar element ratio of sulfur to gadolinium (1, 32) results in an average degree of loading of the polymer of 43.4% Gd-Gly-Me-DOTA and 56.6% ß-D-glucose (corresponding to 28 Gd-Gly-Me -DOTA units and 36 ß-D-glucose residues per molecule). No free amino functions can be detected in the title compound by means of a quantitative ninhydrin reaction.

Example 14

64-DSM-polyamine (Gd-Gly-Me-DOTA amide) 28 - (- ß-D-galactopyranosyl-1-thio-3-propionic acid amide) 36 conjugate

a) 64-DSM-polyamine- (Gly-Me-DOTA-tri-tert-butyl ester amide) 28 - [(3- (2,3,4,6-tetra-O-acetyl) -1 -thio- ß -D-galactopyranosyl-3-propionic acid amide) 36 conjugate

587 mg (5.8 mmol) of triethylamine are added to a stirred solution of 3.0 g (0.132 mmol; corresponding to 3.96 mmol of free amine functions) of the title compound from Example 11e) in 100 ml of absolute dichloromethane at room temperature . Then with a solution of 2.8 g (5.28 mmol) of 3- (2, 3,4,6-tetra-O-acetyl-1-thio-ß-D-galactopyranosyl) -propionic acid-N-hydroxysuccini- midester (representation in analogy to: J. Haensler et al., Bioconjugate Chem. 4, 85, (1993); Chipowsky, S., and Lee.YC (1973), Synthesis of 1 -thio- aldosides; Carbohydrate Research 31, 339-346) in 60 ml dichloromethane added dropwise. After a reaction time of 12 hours at

The room temperature is made up to a total volume of 500 ml with dichloromethane. The organic phase is washed successively twice with water and twice with saturated sodium bicarbonate solution and dried over sodium sulfate. After filtering, the solvent is removed to dryness in vacuo. 4.44 g (89.4% of theory) of the title compound are obtained as a colorless oil. The quantitative ninhydrin reaction of the The title compound is negative, ie no free amino functions can be detected in the polymer.

Eggs entaranaiy se: calc .: C 58.69 H 8.51 N 9.67 S 3.07 found: C 58.58 H 8.62 N 9.80 S 3.02

An average degree of loading of the polymer with 56.5% ß-D-galactose tetraacetate (corresponding to 36 ß-D-galactose residues per molecule) results from the percentage sulfur value of the elemental analysis of the title compound.

b) 64-DSM-Polyamiπ- (Gly-Me-DOTA amide) 28- (ß -D-galactopyranosyl-1 ^• thio-3-propionic acid amide) 36 conjugate

4.0 g (0.10 mmol) of the title compound from Example 14a) are dissolved in 100 ml of absolute dichloromethane. A total of 75 ml of trifluoroacetic acid is then added dropwise at 0 ° C. After a reaction time of 12 hours at room temperature, the mixture is evaporated to dryness in vacuo. The remaining residue is mixed with 100 ml of water and again in

Vacuum drawn to dryness. The residue is dissolved in 100 ml of methanol and 50 ml of a 32% aqueous ammonia solution are added at room temperature. After a reaction time of 12 hours at room temperature, the solvent is removed in vacuo and the remaining residue is dissolved in 200 ml of distilled water. The aqueous product solution is extracted twice with 60 ml of diethyl ether each time. The aqueous product solution is mixed with 10% aqu. Hydrochloric acid brought to pH 3.0, made up to a total volume of 800 ml with water and ultrafiltered three times using a YM3 ultrafiltration membrane (Amicon) against distilled water. The remaining residue is made up to a volume of 500 ml with deionized water and freeze-dried. Yield: 2.3 g (80.36% of theory) as an amorphous powder Water content: 5.44%

Elemental analysis (calculated on anhydrous substance): calc .: C 50.68 H 7.76 N 13.12 S 4.01 found: C 50.91 H 7.90 N 13.10 S 3.98 c) 64-DSM-polyamine (Gd-Gly-Me-DOTA amide) 28 - (- ß-D-galactopyranosyl-1-thio-3-propionic acid amide) 36 conjugate

2.0 g (0.07 mmol; corresponding to 1.94 mmol of Gly-Me-DOTA) der

The title compound from Example 14b) is dissolved in 50 ml of distilled water and the pH of the solution is adjusted to 2.0 by dropwise addition with 10% aqueous hydrochloric acid. Then 520 mg (1.98 mmol) of gadolinium chloride are added at room temperature and the reaction solution is stirred at 60 ° C. for 12 hours. At room temperature, the reaction solution is brought to pH 7.2 by adding 1 molar sodium hydroxide solution and brought to a total volume of 800 ml with distilled water. After three times ultrafiltration against distilled water over a YM3 ultrafiltration membrane (Amicon), the remaining residue is freeze-dried. 2.20 g (95.3% of theory) of the title compound are obtained as an amorphous, colorless powder with a water content of 6.38%.

Elemental analysis (calculated on anhydrous substance): calc .: C 44.08 H 6.50 N 11, 41 S 3.48 Gd 13.29 found: C 43.86 H 6.39 N 11, 48 S 3.65 Gd 13.14

The molar element ratio of sulfur to gadolinium (ratio: 1.36) gives an average degree of loading of the polymer of 42.2% Gd-Gly-Me-DOTA and 57.8% ß-D-galactose (corresponding to 27 Gd-Gly -Me-DOTA units and 36 ß-D-galactose residues per molecule). No free amino functions can be detected in the title compound by means of a quantitative ninhydrin reaction.

Example 15

32-DSM-polyamine- (Gd-DTPA-amide) -i4- (α-D-mannopyranosyl-1-thiopropionic acid amide) -i 8- conjugate [as sodium salt]

a) 32-DSM polyamine (DTPA-tetra-tert-butyl ester amide) - | 4 20.0 g (32.37 mmol) of 3,9-bis (t-butoxycarbonylmethyl) -6-carboxymethyl-3,6,9-triazaundecanedicarboxylic acid di-t-butyl ester (preparation as in the European patent applications: EP 0430863; EP 0331616 and EP 0271180) are at room temperature in 100 ml of absolute

Dissolved dimethylformamide and mixed with 4.47 g (38.84 mmol) of N-hydroxysuccinimide with stirring. The clear reaction mixture is then cooled to 0 ° C. and a total of 7.97 g (38.84 mmol) of dicyclohexylcarbodiimide is added at this temperature in portions. After a reaction time of 30 minutes at 0 ° C. and 2.5 hours at room temperature, the precipitated dicyclohexylurea is filtered off. The filtrate obtained in this way is now slowly added dropwise at room temperature to a stirred solution of 6.7 g (1.9 mmol; corresponding to 60.8 mmol of free amino functions) 32-DSM polyamine and 6.5 g (64.7 mmol) Triethylamine in 60 ml of absolute DMF. After a reaction time of 12 hours at room temperature, the mixture is evaporated to dryness in vacuo and 250 ml of dichloromethane are added to the remaining residue. After filtering off insolubles, the organic phase is washed with saturated sodium bicarbonate solution and twice with water. After the organic product phase has dried over sodium sulfate, the mixture is filtered and the solvent is stripped off in vacuo. You get

18.1 g (80.4% of theory; based on the polyamine used) of a slightly yellow-colored oil, which is used immediately for the subsequent synthesis steps without further purification. After quantitative ninhydrin reaction of the above-mentioned title compound, an average of 18 free amine functions per molecule are present.

Elemental analysis: calc .: C 60.01 H 10.04 N 12.22 found: C 60.13 H 9.91 N 12.34

b) 32-DSM-polyamine-DTPA-tetra-tert-butyl ester amide) - | 4 - [(3- (2,3,4,6-tetra-O-acetyl) -1 -thio-α-D- manπopyranosyl-3-propion-amide] - | Q

A stirred solution of 3.5 g (0.29 mmol; corresponding to 5.3 mmol of free amine functions) of the title compound from Example 15a) in 100 ml of absolute Dichloromethane is mixed with 820 mg (8.1 mmol) of triethylamine at room temperature. Then with a solution of 3.97 g (7.49 mmol) of 3- (2, 3,4,6-tetra-O-acetyl-1-thio-α-D-mannopyranosyl) propionic acid N-hydroxysuccinimide ester ( Lee, YC et al., Biochemi., Vol. 15, No. 18, 1976, 3956-3963; Krohn, KA et al., J. Nucl. Med., Vol. 26, 10, 1985, 1157-1167) added dropwise in 60 ml of dichloromethane. After a reaction time of 12 hours at room temperature, the mixture is made up to a total volume of 500 ml with dichloromethane. The organic phase is washed successively twice with water and twice with saturated sodium bicarbonate solution and dried over sodium sulfate. After filtering, the solvent is removed to dryness in vacuo. 5.06 g (89.4% of theory) of the title compound are obtained as a colorless oil. The ninhydrin reaction of the title compound is negative, ie no free amino functions can be detected in the polymer.

Elemental analysis: calc .: C 56.13 H 8.29 N 7.48 S 2.96 found: C 56.35 H 8.40 N 7.36 S 2.89

An average degree of loading of the polymer with 56% D-mannose tetraacetate (corresponding to 18 D-mannose residues per molecule) results from the percentage sulfur value of the elemental analysis of the title compound.

c) 32-DSM polyamine (DTPA amide) - | 4- (α-D-mannopyranose-1-thio-propionamide) - _| 8-conjugate [as sodium salt]

5.84 g (0.30 mmol) of the title compound from Example 15b) are dissolved in 100 ml of absolute dichloromethane. A total of 75 ml of trifluoroacetic acid is then added dropwise at 0 ° C. After a reaction time of 12 hours at room temperature, the mixture is evaporated to dryness in vacuo. The remaining residue is mixed with 100 ml of water and stripped again to dryness in vacuo. The residue is dissolved in 100 ml of methanol and 50 ml of a 32% aqueous ammonia solution are added at room temperature. After a reaction time of 12 hours at room temperature, the solvent is removed in vacuo and the remaining residue is dissolved in 200 ml of distilled water. The watery Product solution is extracted twice with 60 ml of diethyl ether each, then by adding 10% aqueous solution. Hydrochloric acid brought to pH 3.0, made up to a total volume of 800 ml with water and ultrafiltered three times using a YM3 ultrafiltration membrane (Amicon) against distilled water. After renewed ultrafiltration, the remaining residue is made up to a volume of 500 ml with deionized water and freeze-dried. It is then taken up in 300 ml of water and the pH of the resulting product solution is adjusted to 7.2 by adding 1 molar sodium hydroxide solution. Yield: 3.16 g (87.0% of theory) as an amorphous and colorless powder. Water content: 5.89%

Elemental analysis (calculated on anhydrous substance): calc .: C 42.48 H 4.48 N 6.91 Na 10.59 S 4.75 found: C 42.22 H 4.56 N 7.03 Na 10.23 S 4.70

The ninhydrin reaction of the title compound is negative, i. H. free amino functions are no longer detectable in the polymer.

d) 32-DSM-polyamine- (Gd-DTPA-amide) i4- (α-D-mannopyranose-1-thio-3-propionamide) ι Q [as sodium salt]

2.67 g (0.22 mmol; corresponding to 3.08 mmol of DTPA) of the title compound from Example 15 c) are dissolved in 70 ml of distilled water and the pH of the solution is adjusted to 5 by dropwise addition with 10% aqueous hydrochloric acid. 2 set. Finally, a solution of 815 mg (3.10 mmol) of gadolinium chloride, dissolved in 10 ml of distilled water, is added and the reaction solution is stirred for 12 hours at room temperature. The reaction solution is brought to pH 7.2 by adding 1 molar sodium hydroxide solution and made up to a total volume of 500 ml with distilled water. After three times ultrafiltration against distilled water over a YM3 ultrafiltration membrane (Amicon), the remaining residue is freeze-dried. 2.88 g (98.0% of theory) of the title compound are obtained as an amorphous, colorless powder with a water content of 7.09%.

Elementary anemia (calculated on anhydrous substance): calc .: C 38.66 H 4.06 N 6.27 S 4.31 Gd 16.44 Na 2.48 found: C 38.78 H 4.14 N 6.12 S 4.49 Gd 16.60 Na 2.38

The molar elemental ratios of sulfur to gadolinium thus result in an average degree of loading of the polymer of 44% Gd-DTPA and 56% α-D-mannose (corresponding to 14 Gd-DTPA units and 18 α-D-mannose residues per molecule). No free amino functions can be detected in the title compound by ninhydrin reaction.

Example 16

32-DSM polyamine (Gd-DTPA amide) ι 4- (ß-D-glucopyranosyl-1 ■ thiopropionic acid amide) i 8 conjugate [as sodium salt]

a) 32-DSM polyamine (DTPA-tetra-tert-butyl ester amide) ι 4

10.0 g (16.35 mmol) 3,9-bis (t-butoxycarbonylmethyl) -6-carboxymethyl-3,6,9-triazaundecanedicarboxylic acid di-t-butyl ester (representation as in the European patent applications: EP 0430863; EP 0331616 and EP 0271180) are dissolved in 50 ml of absolute dimethylformamide at room temperature and 2.23 g (19.42 mmol) of N-hydroxysuccinimide are added with stirring. The clear reaction mixture is then cooled to 0 ° C. and a total of 3.99 g (19.42 mmol) of dicyclohexylcarbodiimide is added at this temperature in portions. After a reaction time of 30 minutes at 0 ° C. and 2.5 hours at room temperature, the precipitated dicyclohexylurea is filtered off. The filtrate obtained in this way is then slowly added dropwise at room temperature to a stirred solution of 3.35 g (0.95 mmol; corresponding to 30.4 mmol of free amino functions) 32-DSM polyamine and 3.25 g (32.35 mmol) Triethylamine in 30 ml of absolute DMF. After a reaction time of 12 hours at room temperature, the mixture is evaporated to dryness in vacuo and 150 ml of dichloromethane are added to the remaining residue. After filtering off insolubles, the organic phase is washed with saturated sodium bicarbonate solution and twice with water. After drying the organic

The product phase over sodium sulfate is filtered and the solvent removed in vacuo. 8.87 g (78.8% of theory; based on the amount used Polyamine) of a light yellow colored oil, which is used immediately without further purification for the subsequent synthesis steps. After quantitative ninhydrin reaction of the above-mentioned title compound, an average of 18 free amine functions per molecule are present.

Elemental analysis: calc .: C 60.01 H 10.04 N 12.22 found: C 60.20 H 10.12 N 12.38

b) 32-DSM-polyamine-DTPA-tetra-tert-butyl ester amide) i4 - [(3- (2,3,4,6-tetra-O-acetyl) -1 -thio-ß-D-glucopyranosyl- 3-propion-amide] - | 8

A stirred solution of 3.5 g (0.29 mmol; corresponding to 5.3 mmol of free amine functions) of the title compound from Example 16a) in 100 ml of absolute dichloromethane is mixed with 820 mg (8.1 mmol) of triethylamine at room temperature. Then with a solution of 3.97 g (7.49 mmol) of 3- (2, 3,4,6-tetra-O-acetyl-1-thio-ß-D-glucopyranosyl) propionic acid N-hydroxysuccinimide ester ( Analog representation as described in: Lee, YC et al., Biochemi., Vol. 15, No. 18, 1976, 3956-3963; Krohn, KA et al., J. Nucl. Med., Vol. 26, 10, 1985, 1157-1167; J. Haensler et al., Bioconjugate Chem. 4, 85, (1993); Chipowsky, S., and Lee. YC (1973), Synthesis of 1-thio-aldosides; Carbohydrate Research 31, 339 -346) added dropwise in 60 ml of dichloromethane. After a reaction time of 12 hours at room temperature, the mixture is made up to a total volume of 500 ml with dichloromethane. The organic phase is washed successively twice with water and twice with saturated sodium bicarbonate solution and dried over sodium sulfate. After filtering, the solvent is removed to dryness in vacuo. 4.94 g (87.3% of theory) of the title compound are obtained as a colorless oil. The ninhydrin reaction of the title compound is negative, i. H. free amino functions are no longer detectable in the polymer.

Elemental analysis: calc .: C 56.13 H 8.29 N 7.48 S 2.96 found: C 56.04 H 8.33 N 7.40 S 2.90 An average degree of loading of the polymer with 56% ß-D-glucose tetraacetate (corresponding to 18 ß-D-glucose residues per molecule) results from the percentage sulfur value of the elemental analysis of the title compound.

c) 32-DSM-polyamine- (DTPA-amide) i4- (ß-D-glucopyranose-1-thio-propionamide) - | 8-conjugate [as sodium salt]

5.84 g (0.30 mmol) of the title compound from Example 16b) are dissolved in 100 ml of absolute dichloromethane. Then at 0 ° C with a total

75 ml of trifluoroacetic acid are added dropwise. After a reaction time of 12 hours at room temperature, the mixture is evaporated to dryness in vacuo. The remaining residue is mixed with 100 ml of water and stripped again to dryness in vacuo. The residue is dissolved in 100 ml of methanol and 50 ml of a 32% aqueous ammonia solution are added at room temperature. After a reaction time of 12 hours at room temperature, the solvent is removed in vacuo and the remaining residue is dissolved in 200 ml of distilled water. The aqueous product solution is extracted twice with 60 ml of diethyl ether, then by adding 10% aqu. Hydrochloric acid brought to pH 3.0, made up to a total volume of 800 ml with water and ultrafiltered three times using a YM3 ultrafiltration membrane (Amicon) against distilled water. After renewed ultrafiltration, the remaining residue is made up to a volume of 500 ml with deionized water and freeze-dried. It is then taken up in 300 ml of water and the pH of the resulting product solution is adjusted to 7.2 by adding 1 molar sodium hydroxide solution. Yield: 3.16 g (87.0% of theory) as an amorphous and colorless powder.

Water content: 5.89%

Elemental analysis (calculated on anhydrous substance): calc .: C 42.48 H 4.48 N 6.91 Na 10.59 S 4.75 found: C 42.36 H 4.50 N 6.98 Na 10.41 S 4.78

The ninhydrin reaction of the title compound is negative, ie no free amino functions can be detected in the polymer. d) 32-DSM-polyamine- (Gd-DTPA-amide) -ι - (ß-D-glucopyranose-1-thio-3-propionamide) -] 8 [as sodium salt]

2.67 g (0.22 mmol; corresponding to 3.08 mmol of DTPA) of the title compound from Example 16 c) are dissolved in 70 ml of distilled water and the pH of the solution is adjusted to 5 by dropwise addition with 10% aqueous hydrochloric acid. 2 set. A solution of 815 mg (3.10 mmol) of gadolium chloride in 10 ml of distilled water is then added at room temperature and the reaction solution is stirred for 12 hours at room temperature. The reaction solution is brought to pH 7.2 by adding 1 molar sodium hydroxide solution and made up to a total volume of 500 ml with distilled water. After three times ultrafiltration against distilled water over a YM3 ultrafiltration membrane (Amicon), the remaining residue is freeze-dried. 2.79 g (95.0% of theory) of the title compound are obtained as an amorphous, colorless powder with a water content of 8.96%.

Elemental analysis (calculated on anhydrous substance): calc .: C 38.66 H 4.06 N 6.27 S 4.31 Gd 16.44 Na 2.48 found: C 38.60 H 4.06 N 6.22 S 4.21 Gd 16.54 Na 2.34

The molar elemental ratios of sulfur to gadolinium thus result in an average degree of loading of the polymer of 44% Gd-DTPA and 56% β-D-glucose (corresponding to 14 Gd-DTPA units and 18 β-D-glucose residues per molecule). No free amino functions can be detected in the title compound by ninhydrin reaction.

Example 17

32-DSM-polyamine- (Gd-DTPA-amide) -i3- (α-D-manπopyraπose-1-thio-3-propion-amide) -] g conjugate [as sodium salt]

a) 32 DSM polyamine (DTPA tetra-t-butyl ester amide) - _| 3-conjugate

ERSÄTZBLÄΓT (RULE 26) 25.0 g (40.46 mmol) 3,9-bis (t-butoxycarbonylmethyl) -3-carboxymethyl-3,6,9-triazaundecanedicarboxylic acid di-t-butyl ester (preparation as in the European patent applications: EP 0430863; EP 0331616 and EP 0271180) are dissolved in 100 ml of absolute dimethylformamide at room temperature and 5.58 g (48.50 mmol) of N-hydroxysuccinimide are added with stirring. The clear reaction mixture is then cooled to 0 ° C. and a total of 10.0 g (48.50 mmol) of dicyclohexylcarbodiimide is added at this temperature in portions. After a reaction time of 30 minutes at 0 ° C and 2.5 hours at room temperature, the precipitated dicyclohexylurea is filtered off. The filtrate obtained in this way is slowly added dropwise at room temperature to a stirred solution of 9.0 g (2.56 mmol; corresponding to 82.0 mmol of free amino functions) 32-DSM polyamine and 8.2 g (82.0 mmol) Triethylamine in 100 ml of absolute DMF. After a reaction time of 12 hours at room temperature, the mixture is evaporated to dryness in vacuo and 300 ml of dichloromethane are added to the remaining residue. After filtering off insolubles, the organic phase is washed with saturated sodium bicarbonate solution and twice with water. After drying the organic product phase over sodium sulfate, the mixture is filtered and the solvent is stripped off in vacuo. 25.1 g (86.7% of theory; based on the polyamine used) of a slightly yellow-colored oil are obtained, which is immediately used for the subsequent synthesis steps without further purification. After a quantitative ninhydrin reaction of the above-mentioned title compound, an average of 19 free amine functions per molecule are present.

Elemental analysis: calc .: C 60.95 H 9.99 N 12.51 found: C 61.04 H 10.06 N 12.58

b) 32-DSM-Polyamiπ- (DTPA-tetra-tert-butyl ester -amide) ₁ 3 - [(3- (2,3,4,6-tetra-O-acetyl) -1 -thio-α-D- maπnopyraπosyl-3-propion-amide) - | g conjugate

A stirred solution of 4.0 g (0.35 mmol; corresponding to 6.72 mmol of free amine functions) of the title compound from Example 17a) in 100 ml of absolute dichloromethane is mixed with 815 mg (8.0 mmol) of triethylamine at room temperature. Then with a solution of 3.91 g (7.4 mmol) of 3- (2, 3,4,6-

E - RSARZBLATT (RULE 21) Tetra-O-acetyl-1-thio-α-D-mannopyranosyl) propionic acid N-hydroxysuccinimide ester (Lee, YC et al., Biochemi., Vol. 15, No. 18, 1976, 3956-3963; Krohn , KA et al., J. Nucl. Med., Vol. 26, 10, 1985, 1157-1167) in 100 ml of dichloromethane. After a reaction time of 12 hours at room temperature, the mixture is made up to a total volume of 500 ml with dichloromethane. The organic phase is washed successively twice with water and twice with saturated sodium bicarbonate solution and dried over sodium sulfate. After filtering, the solvent is removed to dryness in vacuo. 5.82 g (86.4% of theory) of the title compound are obtained as a colorless oil. The ninhydrin reaction of the title compound is negative, ie no free amino functions can be detected in the polymer.

Elemental analysis: calc .: C 55.94 H 8.05 N 7.34 S 3.16 found: C 5.02 H 8.13 N 7.29 S 3.14

An average degree of loading of the polymer with 58% α-D-mannose tetraacetate (corresponding to 19 α-D-mannose residues per molecule) results from the percentage sulfur value of the elemental analysis of the title compound.

c) 32-DSM-polyamine (DTPA amide) ι 3- (α-D-mannopyranose-1-thio-3-propionamide) ι g conjugate [as sodium salt]

5.0 g (0.26 mmol) of the title compound from Example 17b) are dissolved in 120 ml of absolute dichloromethane. A total of 85 ml of trifluoroacetic acid is then added dropwise at 0 ° C. After a reaction time of 12 hours at room temperature, the mixture is evaporated to dryness in vacuo. The remaining residue is mixed with 125 ml of water and again in

Vacuum drawn to dryness. The residue is dissolved in 100 ml of methanol and 50 ml of a 32% aqueous ammonia solution are added at room temperature. After a reaction time of 12 hours at room temperature, the solvent is removed in vacuo and the remaining residue is dissolved in 200 ml of distilled water. The aqueous product solution is extracted twice with 80 ml of diethyl ether each time. The aqueous product solution is then mixed with 10% aqu. hydrochloric acid

Θ - töATZBLÄTT (RULE 26) brought to pH 3.0, made up to a total volume of 800 ml with water and ultrafiltered three times using a YM3 ultrafiltration membrane (Amicon) against distilled water. After renewed ultrafiltration, the remaining residue is made up to a volume of 500 ml with deionized water and freeze-dried. It is then taken up in 300 ml of water and the pH of the resulting product solution is adjusted to 7.2 by adding 1 molar sodium hydroxide solution. Yield: 2.78 g (84.7% of theory) as an amorphous and colorless powder. Water content: 8.94%

Elemental analysis (calculated on anhydrous substance): calc .: C 46.78 H 6.72 N 6.60 S 4.08 Na 8.67 found: C 46.59 H 6.88 N 6.39 S 4.33 Na 8.81

d) 32-DSM-polyamine- (Gd-DTPA-amide) ι 3- (α-D-mannopyranose-1-thio-3-propione-amide) -] g conjugate [as sodium salt]

2.5 g (0.19 mmol; corresponding to 2.57 mmol of DTPA) of the title compound from Example 17c) are dissolved in 70 ml of distilled water and the pH of the solution is adjusted to 5 by dropwise addition with 10% aqueous hydrochloric acid , 2 set. Then a solution of 683 mg (2.60 mmol) of gadolinium chloride in 10 ml of distilled water is added at room temperature and the reaction solution is stirred for 12 hours at room temperature. The reaction solution is brought to pH 7.2 by adding 1 molar sodium hydroxide solution and made up to a total volume of 500 ml with distilled water. After three times ultrafiltration against distilled water over a YM3 ultrafiltration membrane (Amicon), the remaining residue is freeze-dried. 2.80 g (98.3% of theory) of the title compound are obtained as an amorphous, colorless powder with a water content of 6.72%.

Elemental analysis (calculated on anhydrous substance): calc .: C 43.15 H 6.28 N 6.09 S 4.08 Gd 13.68 Na 1.98 found: C 43.22 H 6.34 N 6.14 S 4.17 Gd 13.72 Na 2.05 The molar elemental ratio of sulfur to gadolinium thus results in an average degree of loading of the polymer of 42% Gd-DTPA and 58% α-D-mannose (corresponding to 13 Gd-DTPA units and 19 α-D-mannose residues per molecule). No free amino functions can be detected in the title compound by ninhydrin reaction.

Example 18

32 DSM polyamine (Gd-DTPA amide) - | 3- (ß-D-galactopyranose-1-thio-3-propionamide) ι g conjugate [as sodium salt]

a) 32-DSM polyamine (DTPA tetra tert.butyl ester amide) - | 3 - [(3- (2, 3,4,6-Tetra-O-acetyl) -1 -thio- ß-D-galactopyranosyl-3-propione-amide) - | g conjugate

In an analogous manner as described for Example 17b), the reaction of 5.0 g (0.44 mmol; corresponding to 8.4 mmol of free amine functions) of the title compound from Example 17a) with 4.88 g (9.23 mmol) 3- (2,3,4,6-Tetra-O-acetyl-1-thio-β-D-galactopyranosyl) propionic acid N-hydroxysuccinimide ester (representation according to: Lee.YC et al., Biochemi., Vol. 15 , No. 18, 1976, 3956-3963; Krohn, KA et al., J. Nucl. Med., Vol. 26, 10, 1985, 1157-1167) to form 7.48 g (88.3% of theory) . Th.) Of the above-mentioned title compound as a colorless oil.

Elemental analysis: calc .: C 55, 94 H 8.05 N 7.34 S 3.16 found: C 56.02 H 8.11 N 7.38 S 3.20

The title compound shows a negative ninhydrin reaction. The percentage sulfur value of the elemental analysis gives an average degree of contamination of the polymer with 19 β-D-galactose residues per molecule.

b) 32-DSM-polyamine (DTPA amide) i 3- (ß-D-galactopyranose-1-thio-3-propionamide) conjugate [as sodium salt] In an analogous manner to that described for Example 17c), the deprotection of 7.0 g (0.36 mmol) of the title compound from Example 18a) after freeze-drying leads to the formation of 3.93 g (86.3% of theory) of the above title compound as an amorphous powder.

Water content: 4.58%

Elemental analysis (calculated on anhydrous substance): calc .: C 46.78 H 6.72 N 6.60 S 4.08 Na 8.67 found: C 46.83 H 6.90 N 6.54 S 3.98 Na 8.87

c) 32-DSM-Polyamiπ- (Gd-DTPA amide) - _j 3- (ß-D-galactoρyranose-1-thio-3-propioπ-amide) -] g conjugate [as sodium salt]

In an analogous manner to that described for Example 17d), the complexation of 3.5 g (0.27 mmol; corresponding to 3.60 mmol of DTPA) of the title compound from Example 18b) with 956 mg (3.64 mmol) of gadolinium chloride leads to the formation of the above Title link. Yield: 3.92 g (97.3% of theory) of the title compound as a colorless and amorphous powder

Water content: 7.33%

Elemental analysis (calculated on anhydrous substance): calc .: C 43.15 H 6.28 N 6.09 S 4.08 Gd 13.68 Na 1.98 found: C 43.20 H 6.38 N 6.14 S 4.13 Gd 13.75 Na 2.05

Average degree of loading per molecule of the above-mentioned title compound:

13 Gd-DTPA units and

19 β-D-galactopyranose units. Example 19

32-DSM-Polyamiπ- (Gd-DTPA amide) -i3- (ß-D-glucopyranose-1-thio-3-propion-amide) ι g conjugate [as sodium salt]

a) 32-DSM-polyamine- (DTPA-tetra-tert-butiester-amide) -] 3 - [(3- (2,3,4,6-tetra-O-acetyl) -1-thio-ß-D - Glucopyranosyl-3-propion-amide) -j9-conjugate

In an analogous manner as described for Example 17b), the reaction of 6.0 g (0.53 mmol; corresponding to 10.03 mmol of free amine functions) of the title compound from Example 17a) with 5.83 g (11.03 mmol) 3- (2,3,4,6-Tetra-O-acetyl-1-thio-β-D-glucopyranosyl) propioic acid N-hydroxysuccinimide ester (representation according to: Lee.YC et al., Biochemi., Vol. 15 , No. 18, 1976, 3956-3963; Krohn, KA et al., J. Nucl. Med., Vol. 26, 10, 1985, 1157-1167) to form 8.70 g (85.6% of theory) . Th.) Of the above-mentioned title compound as a colorless oil.

Elemental analysis: calc .: C 55, 94 H 8.05 N 7.34 S 3.16 found: C 55.96 H 8.00 N 7.32 S 3.14

The title association shows a negative ninhydrin reaction. From the percentage sulfur value of the elemental analysis, an average degree of exposure of the polymer with 19 ß-D-glucose residues per molecule results.

b) 32-DSM-polyamine (DTPA amide) ι 3- (ß-D-glucopyranose-1-thio-3-propionamide) -j g-conjugate [as sodium salt]

In an analogous manner to that described for Example 17c), the deprotection of 8.0 g (0.41 mmol) of the title compound from Example 19a) after freeze-drying leads to the formation of 4.54 g (87.6% of theory) of Above title compound as an amorphous powder.

Water content: 3.97%

Elemental analysis (calculated on anhydrous substance): calc .: C 46.78 H 6.72 N 6.60 S 4.08 Na 8.67

SPARE BLADES (RULE 26) found: C 46.70 H 6.91 N 6.33 S 4.25 Na 8.66

c) 32-DSM-polyamine- (Gd-DTPA-amide) i3- (ß-D-glucopyranose-1-thio-3-propione-amide) -] g conjugate

In an analogous manner as described for Example 17d), the complexation of 4.0 g (0.31 mmol; corresponding to 4.11 mmol of DTPA) of the title compound from Example 19b) with 1100 mg (4.20 mmol) of gadolinium chloride leads to the formation the above title compound.

Yield: 4.55 g (96.3% of theory) of the title compound as a colorless and amorphous powder

Water content: 6.82% elemental analysis (calculated on anhydrous substance): calc .: C 43.15 H 6.28 N 6.09 Na 2.00 S 4.08 Gd 13.68 found: C 43.20 H 6.36 N 6.14 Na 2.04 S 4.12 Gd 13.72

Average degree of loading per molecule of the above-mentioned title compound: 13 Gd-DTPA units and

19 β-D-glucopyranose units.

Example 20

32-DSM-polyamine- (Gd-DTPA-amide) - | 8- (0-ß-D-galactopyranosyl- (1- → 4) -D- glucono-1,5-lactoπ-amide) i 4 conjugate [ as sodium salt]

a) 32-DSM-polyamine (DTPA amide) ι 8- (0-ß-D-galactopyranosyl- (1 → 4) -D- glucono-1, 5-lactone amide) i4 conjugate [as sodium Salt ]

1.0 g (0.0835 mmol; corresponding to 1.17 mmol of free amino functions) of the titanium compound from Example 6a) are suspended in 10 ml of absolute dimethyl sulfoxide at room temperature. Then a solution of 2.60 g (7.25 mmol; O-ß-D-galactopyranosyl- (1 → 4) -D-glucono-1, 5-lactone [preparation according to the methods described in the literature: ( a) Williams, T. et al., (b) Kobayashi, K., Sumitomo, H., Ina, Y., Polym. J. 1985, 17, 567, (c) Hiromi Kitano, Katsuko Sohda, and Ayako Kosaka, Bioconjugate Chem. 1995, 6, 131-134] in 20 ml of absolute dimethyl sulfoxide are added dropwise and the reaction solution is stirred at 40 ° C. for 1 h. The above-mentioned title compound is obtained by pouring the reaction solution into 250 ml of absolute methanol with stirring at room temperature, which leads to the formation of the

Title compound leads as a colorless precipitate. The precipitate obtained in this way is filtered off with suction, washed well with methanol and then dissolved in 300 ml of distilled water and filtered through an ultrafiltration membrane (YM-3, Amicon). The remaining residue is taken up in water and freeze-dried.

Yield: 1.22 g (87.2% of theory) as a colorless lyophilisate. Water content: 5.98%

Elemental analysis (calculated on anhydrous substance): calc .: C 43.35 H 6.84 N 9.71 Na 9.89 found: C 43.54 H 6.92 N 9.65 Na 10.01

After elemental analysis and colorimetric titration with 0.0025 molar gadolinium sulfate standard solution in the presence of xylene orange as an indicator, the degree of DTPA loading of the polymer is 56.8% (corresponding to 18 DTPA units per molecule). After a quantitative ninhydrin reaction, no free amino functions can be detected in the title compound.

b) 32-DSM-polyamine (Gd-DTPA amide) i 8- (0-ß-D-galactopyranosyl- (1 → 4) - D-glucono-1, 5-lactone-amide) i4 conjugate [as Sodium salt]

0.5 g (0.03 mmol; corresponding to 0.54 mmol of DTPA) of the title compound from Example 20a) are analogous to that described for Example 1c) with 143 mg (0.55 mmol) of gadolinium oxide, dissolved in 7.5 ml distilled water, complexed. After working up and freeze-drying, 0.53 g (98.0% of theory) of the title compound is obtained as an amorphous powder. Water content: 7.23% Elemental analysis (calculated on anhydrous substance): calc .: C 39.59 H 6.26 N 8.87 Gd 15.45 Na 2.26 found: C 39.66 H 6.18 N 8.97 Gd 15.60 Na 2.30

Example 21

64-DSM-polyamine- (Gd-DTPA-amide) 38- (O-ß-D-galactopyranosyl- (1-> 4) -D-glucono-1,5-lactone-amide) 26-conjugate [as sodium Salt ]

a) 64-DSM-polyamine (DTPA amide) 38- (O-ß-D-galactopyranosyl- (1-4) -D-glucono-1, 5-lactone amide) 26 conjugate [as sodium salt ]

1.0 g (0.044 mmol; corresponding to 1.14 mmol of free amino functions) of the title compound from Example 1 a) are suspended in 10 ml of absolute dimethyl sulfoxide at room temperature. Then a solution of 2.55 g (7.12 mmol; O-β-D-galactopyranosyl- (1 → - 4) -D-glucono-1, 5-lactone [preparation according to the methods described in the literature: (a) Williams, T. et al., (b) Kobayashi, K., Sumitomo, H., Ina, Y., Polym. J. 1985, 17, 567, (c) Hiromi Kitano, Katsuko Sohda, and Ayako Kosaka, Bioconjugate Chem. 1995, 6, 131-134] in 20 ml of absolute dimethyl sulfoxide and dropwise the reaction solution for 1 hour at 40 ° C. The above-mentioned title compound is obtained by submerging the reaction solution in 250 ml of absolute methanol at room temperature The precipitate thus obtained is filtered off with suction, washed well with methanol and then dissolved in 300 ml of distilled water and filtered through an ultrafiltration membrane (YM-3, Amicon). The residue which remains is filtered off taken up in water and freeze-dried. Yield: 1.32 g (90.4% of theory . Th.) As a colorless lyophilisate. Water content: 3.92%

Elemental analysis (calculated on anhydrous substance): calc .: C 43.41 H 6.09 N 9.84 Na 10.49 found: C 43.60 H 6.31 N 9.95 Na 10.70 After elemental analysis and colorimetric titration with 0.0025 molar gadolinium sulfate standard solution in the presence of xylene orange as an indicator, the degree of DTPA loading of the polymer is 59.8% (corresponding to 38 DTPA units per molecule). After a quantitative ninhydrin reaction, no free amino functions can be detected in the title compound.

b) 64-DSM-polyamine (Gd-DTPA-amide) 38- (O-ß-D-galactopyranosyl- (1-4) - D-glucono-1, 5-lactone-amide) 26 conjugate [as sodium -Salt ]

1.0 g (0.03 mmol; corresponding to 1.14 mmol of DTPA of the title compound from Example 21 a) are, in analogy to that described for Example 1 c), with 0.30 g (1.15 mmol) gadolinium oxide, dissolved in 10 ml of distilled water, complexed. After working up and freeze-drying, 1.06 g (96.3% of theory) of the title compound is obtained as an amorphous powder. Water content: 7.23%

Elemental analysis (calculated on anhydrous substance): calc .: C 39.44 H 5.45 N 8.94 Gd 16.30 Na 2.38 found: C 39.53 H 5.53 N 9.02 Gd 16.44 Na 2.41

Example 22

64-DSM-polyamine (Gd-Gly Me-DOTA amide) 38 - [1-O- (4-thioureido-tetra-ethylene glycolyl-pheny) -β-D-galactopyranosyl] 26 conjugate

a) 1 -O- [4-nitrophenyl] tetraethylene glycol

863.2 g (5 mol) of tetraethylene glycol are dissolved in 2.5 l of anhydrous toluene, and 159.4 g (2.5 mol) of 88% potassium hydroxide powder and 7.5 g of tetrabutylammonium hydrogen sulfate are added in succession at room temperature. A solution of 53 ml (500 mmol) of 4-fluomitrobenzene in 500 ml of anhydrous toluene is then added dropwise at 0 ° C. with vigorous stirring. After the addition has ended, the mixture is stirred for a further 4 hours at 0 ° C. and for 12 hours at room temperature. For working up, the reaction mixture is stirred into 2000 ml of ice water and then the toluene phase is separated off and the aqueous phase is exhaustively extracted with toluene. After washing the combined organic phases with water, it is dried over sodium sulfate, filtered and the solvent is removed to dryness in vacuo. The remaining, slightly yellowish oily residue is purified by column chromatography (eluent: ethyl acetate / hexane = 3: 1). 135.8 g (86.1% of theory) of a slightly yellowish oil are obtained.

Elemental analysis: calc .: C 53.33 H 6.71 N 4.44 found: C 53.27 H 6.68 N 4.50

b) 2,3,4,6-tetraacetyl-β-D-galactopyranosyl-1-O - [(4-nitrophenyl) - tetraethylene glycol]

10 g of β-D-galactose pentaacetate (commercially available; Aldrich) are dissolved in 40 ml of absolute 1, 2-dichloroethane and with a solution of 8.0 g (25.6 mmol) of the title compound from Example 22a) in 15 ml of 1, 2-dichloroethane added. Then 3.9 ml (33.3 mmol) of tin tetrachloride

Room temperature slowly added dropwise and the reaction solution thus obtained is stirred for 12 h at room temperature. For working up, water is added, the organic phase is separated off and the remaining aqueous phase is exhaustively extracted with dichloromethane. After the organic phases have been dried over sodium sulfate, the mixture is filtered and the solvent is stripped off to dryness in vacuo. The remaining oily residue is purified by column chromatography (eluent: dichloromethanol / methanol = 20: 1). 14.6 g (88.2% of theory) of the above-mentioned title compound are obtained as a colorless oil.

Elemental analysis: calc .: C 52.09 H 6.09 N 2.17 found: C 52.13 H 6.11 N 2.21 c) β-D-galactopyranosyi-1-O - [(4-nitrophenyl) -tetraethyiengiycol]

33.93 g (52.56 mmol) of the title compound from Example 22b) are dissolved in 680 ml of absolute methanol, and 0.8 g (14.8 mmol) of sodium methoxide are added. After a reaction time of 12 hours at room temperature, the reaction solution is neutralized by adding IRA-120- (H ⁺ ) cation exchanger® in portions, suctioned off from the exchanger and the solvent is removed to dryness in vacuo. The title compound thus obtained (23.3 g; 95.5% of theory) is used directly for the subsequent reduction step without further purification.

Elemental analysis: calc .: C 50.31 H 6.54 N 2.93 found: C 50.48 H 6.74 N 3.07

d) β-D-galactopyranosyl-1-O - [(4-aminophenyl) tetraethylene glycol]

The reduction of 14.61 g (30.6 mmol) of the title compound from Example 22c) is carried out according to the procedures described in the literature, [see for example:

Taylor, J.K. et al., J. Chem. Soc, Chem. Commun .; 1993, 1410; Goldstein I.J., Methods in Enzymology, Vol. XXVII, Part B., 212-219, 1972] using hydrogen (1 atm.) As a reducing agent, palladium (10%) on barium sulfate as a catalyst and anhydrous methanol as a solvent. After working up as described in the literature, 13.15 g (96% of theory) of the title compound are obtained as a colorless oil.

Elemental analysis (calculated on anhydrous substance): calc .: C 53.68 H 7.43 N 3.13 found: C 53.43 H 7.60 N 3.20

e) β-D-galactopyranosyl-1 -O - [(4-isothiocyanatophenyl) tetraethylene glycol]

Through implementation known from the literature [cf. J.Goidstein, D.H. Buss, J. Chem. Soc. C, (1968), 1457] of 13.24 g (30.0 mmol) of the title compound from Example 22d) with a total of 5.1 ml of Thiophosgeπ in 850 ml of 80%

SPARE BLADE (RULE 26) aqueous ethanol, 12.62 g (86% of theory) of the above-mentioned title compound is obtained as a colorless oil after working up

Elemental analysis (calculated on anhydrous substance) over C 53.26 H 6.60 N 2 96 S 6.77 found C 53.13 H 6.69 N 3 04 S 6.84

f) 10- [1-Carboxymethylcarbamoyl) ethyl] -1, 4.7, 10-tetraazacyclododecane-1, 4,7-trιessιgsaure

77 g (103.1 mmol) of the title compound from Example 11d) are dissolved in 500 ml of trifluoroacetic acid and stirred for 3 hours at room temperature. The mixture is evaporated to dryness, the residue is taken up in 300 ml of water and the solution is poured onto a column. filled with Reillex® 425 PVP. Eluted with water. The product-containing fractions are combined and evaporated to dryness. The residue is converted from methanol / acetone. Yield 44.04 g (84% of theory) of a colorless, hygroscopic solid, water content 6.5%.

Elemental analysis (calculated on anhydrous substance) over C 47.99 H 6.99 N 14.73 found C 47.83 H 7.12 N 14.55

g) Gadolinium complex of 10- [1-carboxymethylcarbamoyl) -ethyl] -1, 4,7,10-tetraazacyclododecan-1, 4,7-trιessιgsaure

15 27 g (42.06 mmol) of gadolinium oxide are added to 40 g (84.12 mmol) of the title compound from Example 22f), dissolved in 400 ml of water, and the mixture is heated at 90 ° C. for 3 h. It is evaporated to dryness (vacuum) and recrystallizes the residue from 90% aqu ethanol. The crystals are filtered off with suction, washed once with ethanol, then with acetone and finally with diethyl ether and dried in a vacuum oven at 130 ° C. (24 hours). Yield 50.53 g (93% of theory) ) of a colorless crystalline powder water content 2.5% Elemental analysis (calculated on anhydrous substance): calc .: C 36.24 H 4.80 N 11, 12 Gd 24.97 found: C 36.35 H 4.95 N 10.98 Gd 24.80

h) 64-DSM polyamine (Gd-Gly-Me-DOTA amide) 38 conjugate

A stirred suspension of 5.0 g (7.95 mmol) of the title compound from Example 22g) in 15 ml of absolute dimethyl sulfoxide is mixed at 70 ° C. with 0.68 g (15.9 mmol) of lithium chloride. After 30 minutes at 70 ° C., the now clear reaction solution is mixed in portions with a total of 1.83 g (15.9 mmol) of N-hydroxysuccinimide and the reaction mixture is kept at this temperature for 1 hour. After cooling to 13 ° C., 4.52 g (23.85 mmol) of dicyclohexylcarbodiimide are added and the reaction solution is stirred for a further hour at 13 ° C., followed by 12 hours at 22 ° C. The reaction solution of the N-hydroxysuccinimide ester of the title compound from Example 22g) thus obtained is now added dropwise at 22 ° C. with a solution of 1.43 g (0.20 mmol, corresponding to 13.23 mmol of free amine functions), 64-DSM polyamine , dissolved in 15 ml of absolute dimethyl sulfoxide. and stirred for a further 12 hours at room temperature. For working up, the reaction solution is added dropwise in 1.5 l of acetone at 22 ° C., the title compound precipitating out as a colorless precipitate. The precipitate is suctioned off, dissolved in 200 ml of distilled water and ultrafiltered three times over a YM3 ultrafiltration membrane (AMICON ®). The remaining residue is dissolved in 300 ml of distilled water, filtered and freeze-dried.

Yield: 4.0 g (86.0% of theory, based on the polyamine used) of the above-mentioned title compound as an amorphous and colorless solid. Water content: 7.67%

Elemental analysis (calculated on anhydrous substance): calc .: C 37.06 H 5.24 N 11.37 Gd 25.54 found: C 37.13 H 5.37 N 11.22 Gd 25.32

According to elemental analysis, the degree of loading of the polymer is 59.3% Gd-Gly-Me-DOTA (corresponding to 38 Gd-Gly-Me-DOTA units per molecule). The determination of the free amino functions remaining in the polymer by means of The mean value of ninhydrin and the TNBS method is 40.7% of free amino groups, ie there are 26 free amino functions in the polymer molecule.

i) 64-DSM-polyamine- (Gd-Gly Me-DOTA-amide) 38 - [1 -O- (4-thioureido-tetra-ethylene glycolyl-pheny) - ß-D-galactopyranosyl] 26 conjugate

In an analogous manner to that described in Example 1 b), the reaction of 4.0 g (0.17 mmol, corresponding to 4.42 mmol of free amine functions) of the title compound from Example 22h) with 10.92 g (23.0 mmol) of After working up, the title compound from Example 22e) 5.04 g (83.0% of theory, based on the polyamine used) of the above-mentioned title compound as a colorless lyophilisate.

Elemental analysis (calculated on anhydrous substance): calc .: C 42.59 H 5.70 N 7.44 S 2.33 Gd 16.71 found: C 42.38 H 5.84 N 7.63 S 2.43 Gd 16.89

The quantitative ninhydrin reaction as well as the TNBS method are negative, i. H. the polymeric product contains no free amino functions. A degree of loading of the polymer with 26 β-D-galactose residues and 38 Gd-Gly-Me-DOTA complexes per molecule results from the molar sulfur value of the elemental analysis of the title compound.

Example 23

64-DSM-Poiyamin- (Gd-Gly Me-DOTA amide) 38 - [1 -O- (4-thioureidotetraethylene glycolyl-pheny) -α-L-fucopyranosyl] 26 conjugate

a) α-L-Fucopyrannosyl-1 -O - [(4-isothiocyanatophenyl) tetraethylene glycol]

In an analogous manner, as described in Examples 22a) to 22e) for the formation of the title compound from Example 22e), the conversion of 12.76 g of α-L-fucose tetraaacetate (38.4 mmol; commercially available; Sigma) 13, 16 g (72.4% of theory; over 4 synthesis steps) of the above-mentioned title compound as a colorless oil. Elemental analysis (calculated on anhydrous substance): calc .: C 52.26 H 6.60 N 2.96 S 6.77 found: C 52.44 H 6.69 N 3.08 S 6.68

b) 64-DSM-Po! yamin- (Gd-Gly Me-DOTA-Amide) 38 - [1 -O- (4-Thioureido-tetra-ethylene glycolyl-pheny) -α -L-fucopyranosyl] 26 conjugate

The reaction of 5.0 g (0.21 mmol, corresponding to 5.58 mmol of free amino functions), the title compound from Example 22h), with 13.73 g (29 , 0 mmol) of the title compound from Example 23a) to form 6.48 g (86.3% of theory; based on the polyamine used) of the above-mentioned title compound as a colorless lyophilisate.

The quantitative ninhydrin reaction as well as the TNBS method are negative, i. H. the polymeric product contains no free amino functions. A degree of loading of the polymer with 26 α-L-fucose residues and 38 Gd-Gly-Me-DOTA complexes per molecule results from the molar sulfur value of the elemental analysis of the title compound.

Example 24

64-DSM-polyamine (Gd-Gly Me-DOTA amide) 38 - [1 -O- (4-thioureidotetraethylene glycolyl-pheny) -α-D-mannopyranosyl] 26 conjugate

a) α-D-Mannopyrannosyl-1 -O - [(4-isothiocyanatophenyl) tetraethylene glycol]

In an analogous manner, as described in Examples 22a) to 22e) for the formation of the title compound from Example 22e), the reaction of 15 g of α-D-mannose pentaacetate (38.4 mmol; commercially available; Sigma) leads to Formation of 13.22 g (72.4% of theory; over 4 synthesis steps) of the above-mentioned title compound as a colorless oil.

Elemental analysis (calculated on anhydrous substance): calc .: C 53.26 H 6.60 N 2.96 S 6.77 found: C 53.13 H 6.69 N 3.04 S 6.84

b) 64-DSM polyamine (Gd-Gly Me-DOTA amide) 38 - [1 -O- (4-thioureido-tetra-ethylene glycolyl-pheny) -α-D-mannopyranosyl] 26-onjugate

The reaction of 5.0 g (0.21 mmol, corresponding to 5.58 mmol of free amino functions), the title compound from Example 22h), with 13.8 g (29 , 0 mmol) of the title compound from Example 24a) to form 6.48 g (86.3% of theory; based on the polyamine used) of the above-mentioned title compound as a colorless lyophilisate.

The quantitative ninhydrin reaction as well as the TNBS method are negative, i. H. the polymeric product contains no free amino functions. A degree of loading of the polymer with 26 α-D-mannose residues and 38 Gd-Gly-Me-DOTA complexes per molecule results from the molar sulfur value of the elemental analysis of the title compound.

Example 25

64-DSM-polyamine- (Gd-Gly Me-DOTA-amide) 38 - [1 -O- (4-thioureido-tetra-ethylene glycolyl-pheny) -ß-D-glucopyranosyl] 26 conjugate

a) 1 -O- [4-Isothiocyanatophenyl] tetraetylene glycolyl-β-D-glucopyranose In an analogous manner to that described in Examples 22a) to 22e) for the formation of the title compound from Example 22e), the reaction of 8.52 g (21.77 mmol) of β-D-glucose pentaacetate [presentation according to: Rosemann S., et al., Biochemistry, vol. 8, no. 4, 1969, 1351-1359; Prihar, Behrmann, Biochemistry 1973, 12: 997-1000; for analog syntheses cf. also:

Monsigny, M., et al., Biol. Cell., 51, 187 (1984)] to form 8.17 g (79% of theory over 4 synthesis steps) of the above-mentioned title compound as a colorless oil.

Elemental analysis (calculated on anhydrous substance): calc .: C 53.26 H 6.60 N 2.96 S 6.77 found: C 53, 18 H 6.72 N 3.09 S 6.84

b) 64-DSM-polyamine- (Gd-Gly Me-DOTA-amide) 38 - [1 -O- (4-thioureidotetraethylene glycolyl-pheny) - ß-D-glucopyranosyl] 26 conjugate

By carrying out the reaction in a manner analogous to that described in Example 1b), the reaction of 4.0 g (0.17 mmol, corresponding to 4.42 mmol of free amine functions) of the title compound from Example 22h) with 10.92 g (23.0 mmol) of the title compound from Example 25a) to form 4.86 g (80.5% of theory, based on the polyamine used) of the above-mentioned title compound as a colorless lyophilisate.

Elemental analysis (calculated on anhydrous substance): calc .: C 46.19 H 5.95 N 9.58 S 2.82 Gd 9.87 found: C 46.30 H 5.90 N 9.69 S 2.66 Gd 9.78

Example 26

Lymph node enrichment after i.v. injection

The title compound from Example 2 [64-DSM-Polyamiπ- (Gd-DTPA) 38- [1- (4-thioureidophenyl) -α-D-mannopyranosyl] 26-conjugate] is in a dosage of 200 μmol Gd / kg body weight in Rats injected. To different At times after the substance application, the Gd concentration in the tissues of the mononuclear phagocytic system (MPS; liver, spleen, mesenteric and peripheral lymph nodes) is determined using ICP-AES.

Gd-yArichment [/ mol II in tissues of the MPS 10 min, 1 h and 24 h after intravenous application of 200 μmol / kg of the title compound from Example 2 (MW + SD, n = 3)

The increase in Gd concentrations is most pronounced in the lymph nodes. From 1 h p.i. the absolute Gd concentration in the lymph nodes is also significantly higher than in the liver and spleen. The substance accumulates similarly well in all lymph node groups, because the Gd concentration is evenly distributed in the mesenteric and peripheral lymph nodes. Thus, the goal of substance accumulation in all lymph node groups is met by this, but also by the other substances according to the invention described here. Due to the extremely high Gd concentration in the lymph nodes 24h p.i. and a very high relaxivity of the substances according to the invention (R1 (in water) = 17.0 L / mmol * sec), these substances can also be used for intravenous MR lymphography with very low doses (> 10 μmol Gd / kg).

Example 27

MR imaging after i.v. injection

The title compound from Example 24 [64-DSM-polyamine (Gd-Gly Me-DOTA amide) 38 - [1 -O- (4-thioureido-tetraethylene glycolyl-pheny) -α-D-mannopyranosyl] 26 conjugate is shown in a dose of 200 μmol Gd / kg body weight injected into rats. The signal influence is examined ex vivo in various lymph node groups.

ERSAΓZBLAΓT (RULE 26) T1-weighted ex vivo MR image (SE 400/15) of lymph node groups embedded in agar after intravenous application of 200 μmol of the title compound from Example 24 in rats (Figures 1 and 2)

The suitability of the substances according to the invention for intravenous MR lymphography is also evident in imaging, because in all examined lymph node groups 24 h p.i. significant enhancement can be observed.

Claims

claims

1. Conjugates consisting of an amino group-containing dendritic polymer, the branches of which are made from vinyl cyanide units, metal ion (s) containing signaling group (s) and mono- or oligosaccharides and optionally cations of inorganic and / or organic bases, amino acids or amino acid amides.

2. Conjugates according to claim 1, characterized in that they are constructed according to the general formula I.

P (K) _m (LZ) _n (I),

where P for a k amino groups - of which (n + m) are each reduced by one hydrogen atom - containing dendritic polymer, the branches of which

Vinyl cyanide units are produced, where (n + m) <k, n and m each for the integers 1 to 149, k for the numbers 12 to 150, K for a signaling group containing metal ion (s), L for a linker and Z represent a mono- or oligosaccharide reduced by one hydroxyl group.

3. Conjugates of the general formula I according to claim 2, characterized in that the sum (n + m) of the signaling groups (K) and the mono- or polysaccharides (Z) bound by L each reduced by one hydroxyl group is equal to the number k of in the polymer-containing amino groups.

4. Conjugates according to claim 3, characterized in that k stands for the numbers 32 or 64.

5. conjugates of the general formula I according to claim 2, characterized in that P for a k amino groups, of which (n + m) each by one

Hydrogen atom are reduced, containing dendritic polymer, the branches of which are made up of acrylonitrile or methacrylonitrile.

6. Conjugates of the general formula I according to claim 2, characterized in that they contain, as signaling group K, a chelate complex consisting of a radical of the general formula II, III, IV, V or VI.

(IN),

wherein

Rl independently of one another is a hydrogen atom or a metal ion equivalent of the elements of atomic numbers 20-32, 37-39, 42-44, 49 or 57-83,

R ^{2 is} a hydrogen atom, a straight-chain or branched C 1 -C 18 -alkyl radical, a phenyl or benzyl radical,

V a -CH2- (CH ₂ ) ₀ - (0) p // \\ group with

o in the meaning of the numbers 0 to 10, p and I in each case in the meaning of the numbers 0 or 1, with the proviso that p only stands for the number 1 if I means the number 1,

T ^{1 is} an -NHCS or -CO group,

U is a -CHR3-CONR3-M ¹ - or -CH ₂ -CH (OH) -M ² - group with R3 independently of one another in the meaning of R ² or the group -CH2- (CH2) ₀ - COOH and Ml and M ² each meaning a phenylene radical or a straight-chain, branched, saturated or unsaturated C-1 -C20-

Alkylene chain which is optionally substituted by 1 -5 (CH2) ₀ -COOH, 1 - 5 OR ² radicals or 1 -8 oxygen atoms, 1 -2-NH-, 1 -2-C (= NH) -, 1 - Contains 5-CONR ³ -, 1 -5-NR ³ CO-, 1 -2 phenylene or 1 -2 phenyleneoxy groups, with the proviso that at least two of the radicals R ^{1 are} metal ion equivalents of the elements of the above-mentioned atomic numbers stand.

7 conjugates according to claim 6, characterized in that M ^" ! For a group

CH ₂ COOH

α CH ₂ ß, α (CH ₂ ) ₂ ß, _α - ff ß, α CH-CH ₂ - ß, α-CH ₂ -CH ₂ -0-CH ₂ -ß, ^^ α - (CH ₂ ) i -2-CO-N-CH ₂ -ß,

CH ₂ COOH

α - CH ₂ -CH ₂ -CO-NH (CH ₂ ) 6-ß, - (CH ₂ ) 5-ß,

stands, where α indicates the linkage to the rest-CONR ³ and ß the linkage to T ¹ .

8 conjugates according to claim 6, characterized in that M ² for a group

CH ₂ COOH

α - CH ₂ ß ■ ^α , _α CH ₂ -NHCO- (CH ₂ ) _{2 3} -ß

- CH, -NHCO-CH, -0-CH, ß

stands, where α indicates the linkage to the rest-CH (OH) - and ß the linkage to T ^" !

9 conjugates according to claim 1, characterized in that they contain as monosaccharides hexoses, pentoses or N-acetyl-neurammosaure, each of which is in the pyranose or furanose form, or their derivatives

10. Conjugates according to claim 1, characterized in that they contain as monosaccharides hexoses, which are in the pyranose form, or their derivatives.

1 1. Conjugates according to claim 9, characterized in that the monosaccharides as 2-amino-2-deoxy, N-acetyl-2-amino-2-deoxy, 6-deoxy, 2-deoxy, 5-carboxy - Or 1-amino-1-deoxy derivatives are present.

12. Conjugates according to claim 1, characterized in that they contain oligosaccharides constructed from the monosaccharides according to claim 9.

13. Conjugates of the general formula I according to claim 2, characterized in that Z represents a mono- or oligosaccharide reduced by one hydroxyl group on the C-1 atom.

14. Conjugates of the general formula I according to claim 2, characterized in that Z represents a mono- or oligosaccharide reduced by one hydroxyl group on the C-2 atom.

15. Conjugates of the general formula I according to claim 2, characterized in that L stands for a group T -M ³ -X, where M ^{3 stands} for a direct bond or has the meaning given for M ¹ , T ² for a - NHCS -, -C (= NH) -, -CH2 or CO group and bind to P, X stands for NH, CO, O and S and bind to Z.

16. Conjugates according to claim 15, characterized in that M ³ for a direct bond or for the groups

δ

- (CH ₂ CH ₂ O) ₁ . ₄ - CH ₂ - (CH ₂ ) _0.1 - _δ , γ_ (CH ₂ ) ₂ -CONH- (CH ₂ ) ₆ -δ_

CH (OH) CH ₂ OH

y— CH-CH (OH) -CH (OH) -δ, γ - (CH ₂ ) 2-CO- (CH ₂ ) 3-δ

stands, where γ indicates the junction at X and δ the junction at T ² .

17. A process for the preparation of conjugates of the general formula I according to claim 1,

P (K) _m (LZ) _n (I),

wherein

P for a k amino groups - of which (n + m) are each reduced by one hydrogen atom - containing dendritic polymer, the branches of which are made from vinyl cyanide units, where (n + m) <k, n and m each for integers 1 to 149, k for the numbers 12 to 150,

K as a signaling group containing a magnetic metal ion (s) for a chelate complex consisting of a radical of the general formula II, III, IV, V or VI

- OC -x - \ / - \ ^ - COOR ¹

N N N

R ¹ OOC - ^ l ^ - COOR ¹

COOR ¹

(N),

R'OOC - ^ / - \ / - \ / - COOR ¹

NNNR ^ OC - ₂ ^ ^ COOR ¹

R C = O

(Holy),

wherein

R ¹ independently of one another is a hydrogen atom or a metal ion equivalent of the elements of atomic numbers 20-32, 37 - 39, 42 - 44, 49 or 57-83, R ^{2 is} a hydrogen atom, a straight-chain or branched C- | -C7-alkyl- residue, a phenyl or benzyl residue,

V a -CH2- (CH ₂ ) ₀ - (0) _p / A group with

o in the meaning of the numbers 0 to 10, p and I each have the meaning of the numbers 0 or 1, with the proviso that p only stands for the number 1 if I means the number 1, T ^{1 is} an -NHCS or -CO group,

U is a -CHR ³ -CONR ³ -M ¹ - or -CH2-CH (OH) -M ² - group with R ³ independently of one another in the meaning of R ² or the group -CH2- (CH2) o-COOH and M ^" ! and M ² each mean a phenylene radical or a straight-chain, branched, saturated or unsaturated C -] - C20 alkylene chain, which is optionally substituted by 1-5 (CH2) o-COOH, 1-5 OR ² - residues or 1-8 oxygen atoms, 1-2-NH-, 1-2-C (= NH) -, 1-5-CONR ³ -, 1-5- NR ³ CO-, 1 -2 phenylene or 1 -2 contains phenyleneoxy groups, with the proviso that at least two of the radicals Rl stand for metal ion equivalents of the elements of the above-mentioned atomic numbers, L for a linker and

Z represents a mono- or oligosaccharide reduced by one hydroxyl group, characterized in that a) a polymer-complex (former) conjugate of the general formula VII

P (K ^, ) _m (VII), where K 'is a radical of the general formula II', 111 ', IV, V or VI'

RO

R ^r OOOC - _Λ / - \ / - \ ^ COOR ^{1 '} NNN

R ^{1 *} OOC - ^y I ^ -COOR ^{1 '}

R ² "^ C = O

¹ dir),

(VI ')

wherein R ¹ 'independently of one another represents a hydrogen atom, an acid protecting group or a metal ion equivalent of the elements of atomic numbers 20-29, 39, 42, 44 or 57-83 and R ² , V, o, p, T and U have the meaning given above to have, L ^* -Z '(VIII),

where n '= 1-3 n, Z' has the meaning given for Z, but the carboxy, amino and hydroxyl groups which may be present in Z are optionally protected, L ^* for a group T ² '-M ³ ' -X With

T ² 'in the meaning of -NCS, -C (= NH) OCH3, - CHO, a -CO-Fg group where

-O-CO-R4 with R ⁴ meaning a straight-chain or branched C- | -C7-alkyl chain, M ³ 'has the meaning given for M ³ , but the carboxy groups optionally contained in M ³ are optionally protected, and X represents NH, CO, O and S, or

b) a polymer saccharide conjugate of the general formula IX

P (L'-Z ') _n (IX),

where L 'has the meaning given for L, but the carboxy groups which may be present in L are optionally protected and P, Z' and n have the meaning given above, with m 'complexes or complexing agents K ^* . where m '= 1-3 m and K ^* for compounds of the general formulas

R ^r OOC- / - \ / - ^ / -COOR ^{1 '}

N N N

R ^r OOC- ^/ ϊ ^ -COOR ^{1 '}

R ^ = 0 i

Fg

I "),

(VI ")

wherein R1 '> R ² , V, U and Fg have the meaning given above, and T' stands for -NCS, -CO-Fg, -C (= NH) OCH ₃ or -CHO or

c) a polymer of the general formula X

P- (H) k (X),

wherein P and k have the meanings given above, in a one-pot reaction with n 'mono- or oligosaccharides of the general formula VIII

L ^* -Z '(VIII),

wherein n ', L ^* and Z' have the meanings given above, with m 'complexes or complexing agents K ^* . where K ^* for compounds of the general form

Fg (III "),

(VI ")

in which m ', R ¹ ', R ² , V, T ¹ ', U and Fg have the meaning given above, the metal ion introduction which may be necessary being carried out before or after the splitting off of the protective groups which may be necessary, with the proviso that if R1 'stands for protective groups, these must be split off before the metal ion is introduced.

18. A pharmaceutical composition comprising at least one conjugate according to claim 1, optionally with the additives customary in galenics.

19. Diagnostic agent containing at least one conjugate according to claim 1, optionally with the additives commonly used in galenics for NMR imaging.

20. Diagnostic agent according to claim 19, characterized in that it is used for NMR lymphography.

21. Diagnostic agent containing at least one conjugate according to claim 1, optionally with the additives common in galenics for X-ray diagnostics.

22. Diagnostic agent containing at least one conjugate according to claim 1, optionally with the additives common in galenics for radio diagnostics and radio therapy.

23. Use of at least one physiologically compatible conjugate according to claim 1 for the production of diagnostic agents for NMR diagnostics.

24. Use of at least one physiologically compatible conjugate according to claim 1 for the production of diagnostic agents for NMR lymphography.

25. A process for the preparation of the pharmaceutical composition according to claim 18, characterized in that one in water or physiological

Bring dissolved or suspended conjugate, optionally with the additives common in galenics, into a form suitable for enteral or parenteral administration.