DE19728954C1 - Saccharide conjugates, pharmaceutical compositions containing them, processes for their preparation and their use - Google Patents

Description

The invention relates to the counter characterized in the claims stand, that is conjugates consisting of an amino group-containing Polymers, 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, these compounds containing pharmaceutical compositions, their use in diagnostics and Process for the preparation of these compounds and agents.

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 clinics and practices, 7th edition, Vol. IV, pp. 475-512, Stuttgart-New York: Georg Thieme Verlag, 1984). The involvement of lymph nodes in a malignancy has been significant influence on the therapeutic measures to be initiated. The most possible Early and accurate diagnosis of lymph node involvement is therefore essential of paramount importance to the patient.

Contrast-enhanced lymphography has been used in medical practice however, so far only unsatisfactory results have been delivered. X-ray examination gene with iodized oils, e.g. B. with iodinated fatty acid esters of poppy oil show on the one hand, good storage capabilities in the lymph nodes and a need bare contrast effect, but on the other hand 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.), Lym phography in malignant tumors, pp. 40-50. Stuttgart: Georg Thieme Verlag, 1976) neither general nor easily applicable. You can, in particular if the application is too fast, in view of the slight vulnerability of the Lymphatic vessels favor lesions. A number of disadvantages are in the oil character of these contrast agents. Since the suspension of the oil first occurs in the blood, the size of the oil droplets cannot be determined in advance. Fall the oil droplets are too large, are often microembolisms, e.g. B. in the lungs, un avoidable. Because the uptake of large oil droplets from the interstitial Space in the lymphatic capillaries is hardly possible, the display succeeds  Lymphatic vessels and lymph nodes, for example after subcutaneous or intra parenchymal injection, only in exceptional cases.

The latter disadvantage is only due to endolymphatic application overcome, but also a previous one, which is also not without complications Dye marking and operative exposure of the lymphatic vessels under local anesthesia must be accepted. Another disadvantage is there too see that the contrast agents based on iodized oils after application can only be eliminated very slowly. Depending on the accessories The form of elimination is weeks to months.

An attempt has been made to emulsify the iodized oils parts to overcome. This achieved the viscosity and droplet size to reduce. In this way the capillary passage could be improved and at Stability of the emulsion can reduce the risk of embolization. This part However, success could only be bought through other disadvantages. Example The effective reduction of the Iodine content per ml of contrast medium and associated inevitable con loss of traction, greater local toxicity at the lymph nodes, hepatotoxic effects and histologically detectable foreign body reaction by the attached Emulsifiers are reinforced.

All contrast media based on iodized oils are not very stable and due to the side effects shown can only be used to a limited extent. Also in EP 0 022 056 derivatives of the triiodinated 5- Aminoisophthalic acid have the ones put into them in medical practice Expectations not met, because after application they come out of the lymph diffuse out the barrel system. Due to their poor storage capacity in these aqueous forms of preparation are at best for the lymph nodes marginally suitable for peripheral lymphography.

Water-soluble iodinated X-ray contrast media such as Iotrolan (Isovist®) have been used for the representation of the lymphatic system is only of limited use area of application. By infusion into the interstitial space (intracutaneous, subcutaneous or intramuscular) are also only those in the infusion catchment area lymphatic vessels or nodes are shown in high contrast. The flood-soluble, low molecular weight substances then diffuse from the lymphatic vessels to the interstitium. This disadvantage cannot either  who compensated by the sensitive method of computer tomography the.

In nuclear medicine, radioactive ^99m 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.

As lymphographics for nuclear magnetic resonance imaging are also superpara magnetic iron oxide particles, e.g. B. AMI-227 (Guimaraes, R., Clement, O., Bittoun, J., Carnot, f., And Frija, G. MR lympho graphy with superparamagnetic iron nanoparticles in rats: pathologic basis for contrast enhancement. At the. J. Roentgenol., 162, 201-207, 1994). The so far on However, animal test results indicate an unsatisfied imaging despite high dosing of the particles.

Various compounds proposed as lymphographics (Hanka, L. et al., Radiology 1996, 198, 365-370) show such animal specificity (rat vs. Guinea pigs and rabbits) that their development is discontinued had to.

The task therefore continued to be to provide connections len, which accumulate in the lymphatic tissue after parenteral application and which Do not have the above disadvantages, e.g. B. compared to the known Structures a more favorable safety distance between diagnostic and have a lethal dose ("margin of safety"). This task is accomplished by the lying invention solved.

It has been found that the conjugates according to the invention consisting of egg nem amino group-containing polymers, metal ion (s) containing si signaling group (s) and mono- or oligosaccharides as well as given if cations of inorganic and / or organic bases, amino acids or Amino acid amides are surprisingly excellent for the production of NMR diagnostics, particularly suitable for lymphography.

Surprisingly, they have the following advantages:

• - Enrichment in the lymph sufficient for good imaging  system,
• - high tolerance (LD ₅₀ ),
• - a short elimination time (they are usually within 14 days more than 98% excreted from the body),
• - their high relaxivity allows them to be applied in low doses,
• - 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 polymer containing a k amino group - of which (n + m) are each reduced by one hydrogen atom - where (n + m) should be ≦ k,
n and m each for the integers 1-149,
k for the numbers 12 to 150,
K for a signaling group containing metal ion (s),
L for a left and
Z represents a mono- or oligosaccharide reduced by one hydroxyl group.

That stands for P, containing k amino groups, of which (n + m) each are reduced by one hydrogen atom, polymer is a dendrimer (e.g. WO 96/01655, EP 0271180), polylysine (e.g. EP 0331616), polypeptide, poly alkylamine, poly [N- (2-aminoethyl)] methacrylamide (e.g. US-5,364,613) or poly saccharide (e.g. EP 0535668). The sum (n + m) of the signal is preferred groups (K) and those bound via L each have a hydroxyl group reduced mono- or polysaccharides (Z) equal to the number k of im Amino groups containing polymers, i.e. H. there is a full cast of the amino groups contained in P. The polymers preferably contain 12 24, 32, 36, 40, 48, 64 or 80 amino groups, of which e.g. B. 3, 4, 6, 7, 8, 11, 13, 14, 16, 18, 19, 21, 47 to signaling groups K and z. B. 9, 8, 18, 17, 16, 13, 11, 10, 20, 6, 29, 27, 33 to each bound by L by one Hy droxyl group reduced mono- or polysaccharides Z are bound.  

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 residue of the general formula II, III, IV, V or VI.

wherein
R ¹ independently of one another is a hydrogen atom or a metal ion equivalent to the elements of the 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 ₇ alkyl radical, a phenyl or benzyl radical,

With
o in the meaning of the numbers 0-10,
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 -CH ₂ -CH (OH) -M ² group with R ³ independently of one another in the meaning of R2 or the group -CH ₂ - (CH ₂ ) _o - COOH and M ¹ and M ² each represent a phenylene radical or a straight-chain, branched, saturated or unsaturated C ₁ -C ₂₀ alkylene chain which is optionally substituted by 1-5 (CH ₂ ) _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 phenyleneoxy groups ent means, 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.

Is the compound of the invention for use in NMR diagnostics determined, the metal ion of the signaling group must be paramagnetic his. These are in particular the divalent and trivalent ions of the elements of the Atomic numbers 21-29, 42, 44 and 58-70. Suitable ions are exemplary show the chrome (III) -, iron (II) -, cobalt (II) -, nickel (II) -, copper (II) -, Praseo dym (III), neodymium (III), samarium (III) and ytterbium (III) ion. Because of her star Magnetic moments are particularly preferred Gadolini um (III) -, terbium (III) -, dysprosium (III) -, holmium (III) -, erbium (III) -, iron (III) - and manganese (II) ions.

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

Is the connection according to the invention for use in X-ray diagnosis stik determined, the metal ion is preferably derived from an element Hö atomic number to ensure adequate absorption of the X-rays to achieve. It has been found that for this purpose diagnostic agents that a physiologically compatible complex salt with metal ions from elements of Contain atomic numbers 25 and 26 as well as 57-83, are suitable.

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

Acidic hydrogen atoms which may be present in R ¹ , that is to say those which have not been substituted by the central ion, can optionally be wholly or partly replaced 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, the calcium ion and especially the sodium ion. Suitable cat Ions of organic bases include those of primary, secondary ones or tertiary amines such as ethanolamine, diethanolamine, morpho lin, glucamine, N, N-dimethylglucamine and in particular N-methylglucamine. Ge suitable cations of amino acids are, for example, those of lysine and Ar ginins and ornithine as well as the amides otherwise acidic or neutral Amino acids.

The compounds of the invention which have a molecular weight of 5,000- 200,000 D, preferably 15,000-60,000 D, have the ge described the desired properties. They contain those for their use The large number of metal ions required in the complex are stably bound.

They accumulate in the lymphatic system and allow statements about perfusion of tissues, give the opportunity to determine the blood volume in tissues and selectively shorten the relaxation times or densities of the blood Zen. Such physiological information is not through the use of extracellular contrast agents such as B. Gd-DTPA [Magnevist®]. The areas of application at mo magnetic resonance imaging and computed tomography phie: more specific diagnosis of metastatic, especially malignant Tumors and early therapy control in cytostatic, anti-inflammatory or vasodilative therapy.

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

As further advantages over extracellular contrast agents, such as. B. Gd- DTPA [Magnevist®], must be more effective as a contrast agent for the core spin tomography (higher relaxivity) are highlighted, resulting in a leads to a significant reduction in the diagnostically necessary dose. At the same time  the contrast media according to the invention can be used as isoosmolar solutions Blood are formulated and thereby reduce the osmotic load on the Body, resulting in a reduced toxicity of the substance (higher toxic Threshold). Lower doses and higher toxic thresholds result to a significant increase in the safety of contrast medium applications in modern imaging processes.

Another advantage of the present invention is that Com plexes with hydrophilic or lipophilic, macrocyclic or open chain, never dermolecular or high molecular weight ligands have become accessible. This gives the possibility of tolerance and pharmacokinetics to control this polymer complex 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 CH ₂ C ₆ H ₄ -, CH ₂ -OC ₆ H ₄ -, (CH ₂ ) ₄ -, (CH ₂ ) ₆ - and (CH ₂ ) _{10 groups} , where the C ₆ H ₄ group is attached to T ¹ .

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

M ¹ may be mentioned as examples

where α indicates the link to the rest -CONR ³ and β the link to T ¹ .

The groups are preferred:

Examples of groups M ² are:

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

The groups are preferred:

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. B. D-Ga lactose, 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-Ga lactose and N-acetyl-neuraminic acid. If Z stands for oligosaccharides, these can be linear or branched from the above-mentioned monosaccharides, which may have been linked terminally or within the chain. Examples of oligosaccharides are: sialyl-Lewis ^x (see BH Ohmoto et al, J. Med. Chem. 1996, 39, 1339); Deoxy-sialyl-Lewis ^x (see BW Stahl et al., Angew. Chemie 1994, 106, 2186); Sialyl-Le ^x (see BW Stahl, J. prak. Chemie 1995, 337, 441); Gentiobiose (Fluka Chemie AG, Switzerland); Lactose; Maltose; Lactulose (Fluka Chemie AG, Switzerland); LactNAc; N, N ', N "-triacetylchitotriose; 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 sugar.

The linker L, via which the mono- or oligosacchaides reduced by one hydroxyl group are bound to P, is written by the group T ² -M ³ -X, where M ^{3 stands} for a direct bond or the meaning given for M ¹ has, T ^{2 stands} for an -NHCS-, -C (= NH) -, -CH ₂ - or CO group and binds to P, X stands for NH, CO, O and S and binds to Z. For example, the groups are M ³

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

The conjugates of the general formula I 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),
  wherein
  K 'for a radical of the general formula II', III ', IV', V 'or 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 ¹ and U have the meaning given above, with n 'mono- or oligosaccharides of the general formula VIII
  L * -Z '(VIII),
  wherein
  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) OCH ₃ , -CHO, a -CO-Fg group
  in which
  Fg for -Cl, -F, -OH,
  -N _{3 '} -O-CO-R ⁴ with R ⁴ meaning a straight-chain or branched C ₁ -C ₇ 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),
  wherein
  L 'has the meaning given for L, but the carboxy groups optionally 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
  wherein
  R ^{1 '} , 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 formulas
  

where m ', R ^1'

, R ²

, V, T ^{1 '}

, and Fg have the meaning given above, is converted, the metal ion introduction which may be necessary taking place before or after the splitting off of the protective groups which may be necessary, with the proviso that if R ^{1 '}

stands for protective groups, these in front of the metal ion introduction must be split off.

If R ^{1 '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 come, for example the methyl, ethyl, propyl, butyl -, Phenyl, Ben zyl, diphenylmethyl, triphenylmethyl, bis (p-nitrophenyl) methyl group, and trialkylsilyl groups in question.

The desired splitting off of the protective groups takes place after the methods known to the person skilled in the art (see, for example, E. Wünsch, Methods der Org. Chemie, Houben-Weyl, Vol XV / 1, 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, acidic ver soaping with mineral acids or in the case of tert-butyl esters with the help of Tri fluoroacetic acid.  

As hydroxyl protective groups such. B. the benzyl, 4-methoxybenzyl, 4-Ni trobenzyl-, trityl-, diphenylmethyl-, trimethylsilyl-, dimethyl-t-butyl-silyl-, di phenyl-t-butysilyl group in question. As a protective group for the 1-OH group of The methyl group is also suitable for mono- and / or oligosaccharides.

The hydroxy groups can also, for. B. as THP ether, alkyl ether, α-Al koxyethyl ether, MEM and MOM ether or as an ester with aromatic or ali phatic carboxylic acids, such as. B. acetic acid and chloroacetic acid, benzoin acid, levulinic acid or 2-chloroacetoxymethyl (or ethyl) benzoic acid (GIT Fachz. Lab. 1996, 46) are available. In the case of polyols, the hydroxy groups also in the form of ketals with z. B. acetone, acetaldehyde, cyclohexanone or be protected by benzaldehyde.

If carboxyl groups are present at the same time, hydroxyl groups also by intramolecular esterification to the corresponding lactones protects present.

The hydroxyl protective groups can be prepared according to the litera known to those skilled in the art tower methods, e.g. B. by hydrogenolysis, reductive cleavage with lithi um / ammonia, acid treatment of ethers and ketals or alkali treatment the esters are released (see, for example, "Protective Groups in Organic Synthe sis ", T. W. Greene, John Wiley and Sons 1981).

The amine protecting groups are the benzyloxycar familiar to the person skilled in the art bonyl, tertiary butoxycarbonyl, trifluoroacetyl, fluorenylmethoxycarbonyl, Ben cyl, formyl, 4-methoxybenzyl, 2,2,2-trichloroethoxycarbonyl, phthaloyl, 1,2- Oxazoline, tosyl, dithiasuccinoyl, allyloxycabonyl, sulfate, pent-4-encarbonyl, 2-chloroacetoxymethyl (or -ethyl) benzoyl, tetrachlorophthaloyl, alkyloxycar bonyl groups [Th. W. Greene, P.G.M. Wuts, Protective Groups in Organic Syntheses, 2nd ed, John Wiley and Sons (1991), pp. 309-385, E. Meinjohanns et al, J. Chem. Soc. Pekin Trans 1, 1995, 405; U. Ellensik et al, Carbohydrates Research 280, 1996, 251; R. Madsen et al, J. Org. Chem. 60, 1995, 7920, R. R. Schmidt, Tetrahedron Letters 1995, 5343].

The implementation specified under a) takes place according to the methods familiar to the person skilled in the art, such as, for. B. 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, 131, 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 be described. It is in solvents such as. B. 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 preferred 1 to 14 hours, optionally with the addition of an organic or inorganic base, such as. B. 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, hydrocarbonate. 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. 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 Bomann 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). As a reducing agent z. B. sodium borohydride, sodium cyanoborohydride, lithium borohydride, calcium borohydride, pyridine-BH ₃ , lithium aluminum hydride and zinc borohydride. A catalytic hydrogenation on z. B. palladium or nickel catalyst is suitable.

If Fg stands for an OH group, an in-situ activation with the Coupling reagents known to those skilled in the art, such as, for. B. 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; Y. M. An  gell et al, Tetrahedron Letters 1994, 35, 5981; L.A. Carpino et at, J. Chem. Soc. Commun. 1994, 201; H-O. 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; be performed.

The reaction indicated under b) of the polymer saccharide conjugate of the general formula IX 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 such as, for. Described, for example, in U.S. 5,135,737, H. Takalo et al, Bioconjugate Chem. 1994, 5,278; EP 0430863; EP 0331616; WO 96/01655; EP 0271 180; U.S. 5,364,613; WO 95/17451 and WO 96/02669. It is in solvents such as. B. 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 temperatures within 5 minutes to 72 hours, preferably 1 to 24 hours, particularly preferably 1 up to 14 hours, optionally with the addition of an organic or inorganic base, such as. B. aromatic or aliphatic amines, alkali or alkaline earth metal hydroxides, carbonates or bicarbonates 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, bicarbonate. 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 value is preferably maintained using a pH state. If T ^{2 'is} 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 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). As a reducing agent z. B. sodium borohydride, sodium cyanoborohydride, lithium borohydride, calcium borohydride, pyridine-BH ₃ , lithium aluminum hydride and zinc borohydride. Catalytic hydrogenation is also suitable.

The reactions indicated under a) and b) are preferably carried out with a Excess of the activated saccharide of formula VIII or the activated Complex or complexing agent K *, preferably with a 1.5 to 3 times Excess.

The dropping-in reaction indicated under c) takes place under that under a) and b) carried out reaction conditions by using a polymer of general Formula X implemented with a mixture of m '= 1-3 m K * and n' = 1-3 n L * -Z '. The occupation ratio of the amino groups contained in the polymer becomes controlled by the ratio n ': m'.

The necessary or already according to the above-mentioned implementations a) -c) in K 'or K * metal ion introduction takes place in the manner as z. B. has been disclosed in German Offenlegungsschrift 34 01 052 by the metal oxide or a metal salt (for example the nitrate, acetate, car bonate, chloride or sulfate) of the element of atomic numbers 20-32, 37-39, 42-44, 49, 57-83 in water and / or a lower alcohol (such as methanol, Ethanol or isopropanol) dissolves or suspended and with the solution or Sus the equivalent amount of the complex-forming ligand and then, if desired, acidic hydrogen atoms present in the acid groups by cations of inorganic and / or organic bases, Amino acids or amino acid amides substituted.

Neutralization takes place with the help of inorganic bases (e.g. hydroxides, Carbonates or bicarbonates) of e.g. B. sodium, potassium, lithium, magnesium or calcium and / or organic bases such as primary, secondary där and tertiary amines, such as. B. ethanolamine, morpholine, glucamine, N-Me thyl- and N, N-dimethylglucamine, and basic amino acids, such as. B. lysine, Arginine and ornithine or from amides originally neutral or acidic Amino acids such as B. hippuric acid, glycine acetamide.  

For example, the acid compounds can be used to prepare the complex compounds Complex salts in aqueous solution or suspension as much of the desired Add bases so that the neutral point is reached. The solution obtained can then concentrated to dryness in vacuo. It is often from Advantage, the neutral salts formed by adding water-miscible Solvents such as B. lower alcohols (methanol, ethanol, isopropanol and others), lower ketones (acetone and others), polar ethers (Tetrahydrofuran, dioxane, 1,2-dimethoxyethane and others) and so on easy to isolate and easy to clean crystals. As a special ders advantageous it has proven to be the desired base already during the Add complex formation of the reaction mixture and thereby a procedure save step.

If the acidic complex compounds contain several free acidic groups, then it often makes sense to produce neutral mixed salts that are both inorganic as well as organic cations as counterions.

This can be done, for example, by using the complex-forming ligan the in aqueous suspension or solution with the oxide or salt of that Central ion supplying element and half of those required for neutralization Reacts amount of an organic base, isolates the complex salt formed, it if necessary, cleans and then for complete neutralization with the be required amount of inorganic base added. The order of the bases addition can also be reversed.

The polysaccharide conjugates obtained in this way are purified if after adjusting the pH by adding an acid or base 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. B. 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 oligosacchaids 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 in the form of the free amines or in the form of their salts, e.g. B. as hydrochlorides, hydrobromides or hydrosulfates be used. They are known from the literature (e.g. polylysine: EP 0331 616; Den drimers: WO 96/01655, EP 0271 180, EP 0430863, polypeptide, polyallylamine, Poly [N- (2-aminoethyl)] methacrylamide: US 5,364,613; Polysaccharide: V. D. Nadkarni et al, Carbohydrate Research 1996, 290, 87, E. A. Yates et al, Carbo hydrate Research 1996, 294, 15, D. J. Tyrrell et al, TiPS 1995, 16, 198, JP- Patent 06080705-A).  

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 II'a: see for example EP 263 059,
III ": see for example DE 195 07 822, DE 195 80 858, DE 195 07 819
IV ": see eg WO 91/14459
V ": see, for example, US-5,053,503, WO 96/02669, WO 96/01655, EP 0430863, EP 255471, US-5,277,895, EP 0232751, US-4,885,363
VI ": see e.g. US-4,923,985

The mono- and oligosaccharides of the general formula required as starting materials VIII are also known from the literature or analogously to methods known from the literature available (see also experimental part): s. e.g. B. EP 0 128097; A.J. Jonas et al. Biochem. J. 1990, 268, 41; JP Patent 04211099; D.H. Buss et al, J. Chem. Soc. C. 1968, 1457; C.M. Hilditch et al, J. Appl. Phycol. 1991, 3, 345; M. Monsigny et al, Biol. Cell 1984, 51, 187; E. P. Dubois et al, Bioorganic and Me dicinal Chemistry Letters 1996, 6, 1387; C.R. 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, 106, 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, 107, 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; E. A.L. 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. Yep 1976, 49, 2639; P.W. Austin et al, J. Chem. Soz. C 1965, 1419; H. Paulsen, Angew. Chem. 1990, 102, 851; R. R. 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, R. R. Schmidt et al, Liebigs Ann. Chem. 1983, 1249; H. A. van Doren et al, Carbohydrate Research 1989, 194, 71, F. D. Tropper et al, J. Car  bohydr. Chem. 1992, 11, 751; T. Sugawara et al, Carbohydrate Research 1992, 230, 117, Y.C. Lee et al, Biochemistry 1976, 15, 3956; Blomberg et al, J. Car bohydr. Chem. 1992, 11, 751.

The pharmaceutical compositions according to the invention are produced if in a conventional manner, by the complex of the invention compounds - if necessary with the addition of the additives common in galenics ze - suspended or dissolved in an aqueous medium and then the suspensions sion or solution sterilized if necessary. Suitable additives are for example wise physiologically harmless buffers (such as tromethamine), additions of Complexing agents or weak complexes (such as diethylene tria minpentaacetic acid or the corresponding Ca polysaccharide polymer Complexes) or - if necessary - electrolytes such as sodium chloride or - if necessary - antioxidants such as ascorbic acid.

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

In principle, it is also possible to use the pharmaceutical compositions according to the invention can also be produced without isolating the complex salts. In any case, special whose care is taken to chelate so that the salts and salt solutions according to the invention are practically free from complexed toxic metal ions.

This can be done, for example, with the help of color indicators such as xylenol orange Control titrations can be guaranteed during the manufacturing process. The invention therefore also relates to methods for producing the complex compound and their salts. As a final security, cleaning of the isolated remains 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 of 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 Suitability as a contrast medium for magnetic resonance imaging. So they are outstanding suitable to increase after oral or parenteral application the signal intensity in the image obtained with the aid of an MRI scanner to improve its informative value. They also show the high effectiveness that is necessary to the body with the smallest possible amount of foreign substances strain, and the good tolerance that is necessary to the non-invasi to maintain the nature of the investigations.

The good water solubility and low osmolality of the invention Medium allows to produce highly concentrated solutions, so the volume keep the stress on the circulatory system within reasonable limits and the dilution by balancing the body fluid, that is, NMR diagnostics must be done Be 100 to 1000 times more water-soluble than for 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 a Release or exchange of those not covalently bound in the complexes - itself toxic - ions within the time when the new contrast agent is full are constantly eliminated, takes place extremely slowly.

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

Particularly low doses (below 1 mg / kg body weight) of organ specific NMR diagnostics are for example for the detection of tumors and usable from heart attack.

The agents according to the invention are due to their favorable radioactive egg properties and the good stability of the complex compound contained in them also suitable as radio diagnostics. Details of their application and Thursday sation z. B. in "Radiotracers for Medical Applications", CRC-Press, Boca Raton, Florida.

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

The compounds of the invention are surprisingly also for Differentiation of malignant and benign tumors in areas without blood Brain barrier suitable. They are also characterized by the fact that they are complete dig are eliminated from the body and are therefore well tolerated.

Since the substances according to the invention accumulate in malignant tumors (no diffusion into healthy tissue, but high permeability of tumor vessels), they can also support the radiotherapy of malignant tumors. This differs from the corresponding diagnostics only in the amount and type of isotope used. The goal is the destruction of 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 (e.g. X-rays) or with neutron beams are used for this purpose. 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 and thus adverse side effects for the patient can be avoided. 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, ⁷² Ga, ⁷³ Ga and ⁹⁰ Y. Suitable α-emitting ions with short half-lives are, for example, ²¹¹ Bi, ²¹² Bi, ²¹³ Bi and ²¹⁴ Bi, whereby ²¹² Bi is preferred. A suitable photon and electron emitting ion is ¹⁵⁸ Gd, which can be obtained from ¹⁵⁷ Gd by neutron capture.

In the in vivo application of the therapeutic agents according to the invention these together with a suitable carrier such as serum or physiological saline and along with another pro such as human serum albumin. The Dosie tion depends on the type of cellular disorder, the one used Metal ion and the type of imaging method.

The therapeutic agents according to the invention are preferred parenterally wise i. v., applied.

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

The agents according to the invention are outstanding as X-ray contrast agents suitable, especially for computed tomography (CT), especially here It should be emphasized that they show no signs of iodine-containing Contrast agents known anaphylaxis-like reactions in biochemical pharmacological examinations. Are particularly valuable they because of the favorable absorption properties in areas of higher tubes voltage for digital subtraction techniques.

In general, the agents according to the invention for use as X-ray contrast media in analogy to meglumine diatrizoate in amounts of 0.1 5 mmol / kg, preferably 0.25-1 mmol / kg dosed.

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

Overall, new polymer-metal complexes have been successfully synthesized new opportunities in diagnostic and therapeutic medicine conclude.

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

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;
in which
⇐ 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) DTPA monoanhydride monoethyl ester:

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-tria zaundecanoic acid 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 ester, sodium bromide

7) Gd-Gly-Me-DOTA:

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

8) Sialyl Lewis ^x = [sLe ^x ]: (α-Neu5Ac- (2-3) -β-D-Gal- (1-4) - [a-L-Fuc- (1-3)] - β-GlcNAc)

The polyamines used in the respective syntheses are as follows named once and the corresponding literature for their synthesis cited once:

a) 12-cascade polyamine (12-mer polyamine):

12-polyamine based on the N, N, N ', N', N ", N" - hexakis [2- (trilysylamino) - ethyl] -trimesic acid amide cores with N, N'-dibenzyloxycarbonyl-L-lysine

Synthesis description in an analogous manner as in DE 195 25 924 A1 in example 1d) described for the representation of the 24-cascade polyamine, only under Use of N, N'-dibenzyloxycarbonyl-L-lysine instead of one completely Benzyloxycarbonyl protected "tri-lysine".

b) 24-cascade polyamine (24-mer polyamine):

24-polyamine based on the N, N, N ', N', N ", N" - hexakis [2- (trilysylamino) - ethyl] trimesic acid amides  

Synthesis description in: DE 195 25 924 A1; EP 0430 863 A1; WO 97 / 02051; DE 195 18 222 A1

c) 36-cascade polyamine (36-mer polyamine):

36-polyamine based on the N, N, N ', N', N ", N" - hexakis [2- (trilysylamino) - ethyl] -trimesic acid amide cores with six 1- (carboxymethoxyacetyl) -4,7,10- tris (N, N'-dibenzyloxycarbonyl-lysyl) -1,4,10-tetraazacyclododecane

Synthesis description in: EP 0430 863 A1; WO 97/02051

d) 48-cascade polyamine (48-mer polyamine):

48-polyamine based on 1- (carboxymethoxyacetyl) -4,7,10- tris (N, N'-dibenzyloxycarbonyl-lysyl) -1,4,7,10-tetraazacyclododecane Hexamine cores with 1,4,7,10-tetraazacyclododecane tetrakis (N, N'- dibenzyloxycarbonyl-lysyl)

Synthesis description in: EP 0271180

f) 24 Starburst Dendrimer:

Tomalia D.A. et al .; Polym. J. (Tokyo) 1985, 17, 117; Tomalia D.A. et al .; Macromolecules 1986, 19, 2466; Tomalia D.A., et al .; Macromolecules 1987, 20, 1164; Tomalia D.A. et al., J. Am. Chem. Soc. 1987, 109, 1601; Tomalia D. A. et al .; J. Org. Chem. 1987, 52, 5305.

e) 48er starburst dendrimer:

Tomalia D.A. et al .; Polym. J. (Tokyo) 1985, 17, 117; Tomalia D.A. et al .; Macromolecules 1986, 19, 2466; Tomalia D.A. et al .; Macromolecules 1987, 20, 1164.

example 1 13 kD poly-L-lysine (Gd-DTPA amide) ₄₇ - [1- (4-thioureidophenyl) -β-D-galactopyranosyl] ₃₃ conjugate a) Polyamide from 13 kD poly-L-lysine and DTPA

A solution of 5.0 g (0.38 mmol; corresponding to 30.4 mmol of free amino functions) poly-L-lysine hydrobromide (13 kD, purchased from Sigma) is dissolved in 400 ml of distilled water and a pH of 1 molar sodium hydroxide solution is used. Value set to 9.5. Then a total of 14.7 g (36.5 mmol) of DTPA mono-anhydride mono-ethyl ester (preparation as described in EP 0263051 A1, EP 0450742 A1 and EP 0413405 A1) are added, the pH of the reaction solution being determined by Addition 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, the volume is made up to 800 ml with distilled water. The aqueous product solution thus obtained is cut three times using a YM3 ultrafiltration membrane [cut off 3000 D; AMICON®] ultrafiltered 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: 10.26 g (84.6%; based on the polyamine used) as an amorphous powder
Water content: 7.44%

Elemental analysis (calculated on anhydrous substance):
calc .: C 42.61; H 5.68; N 13.14; Na 13.48;
Found: C 42.55; H 5.72; N 13.20; Na 13.61;

After elemental analysis as well as colorimetric titration in a complex temperature of 22 ° C with 0.0025 molar gadolinium sulfate standard solution in The presence of xylenol orange as an indicator results in a DTPA loading degree of polymer of 58.7% (corresponding to 47 DTPA units per molecule). After a quantitative ninhydrin reaction [cf. see: Chem. Rev., 60, 39 (1960); Tetrahydron, 34, 1285 (1978), 47, 8791 (1991)] as well as 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)] are on average per po polymer molecule 33 free amino functions.

b) 13 kD poly-L-lysine (DTPA amide) ₄₇ - [1- (4-thioureidophenyl) -β-D-galactopyranosyl] ₃₃ conjugate

1.5 g (0.047 mmol; corresponding to 1.54 mmol of free amino functions) of the titanium 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 2.07 g (6.63 mmol) of p-isothiocyanatophenyl-β-D-galactopyranose (representation according to: M. Ponpipom et al., J. Med. Chem. 24, 1388; 1981. General Formation of isothiocyanates from the corresponding aniline precursors of various hexapyranoses 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 1.84 g (92.6% of theory) of an amorphous and colorless solid.
Water content: 6.78%

Elemental analysis (calculated on anhydrous substance):
calc .: C 44.34; H 5.54; N 11.02; S 2.49; Na 10.18;
Found: C 44.27; H 5.70; N 19.97; S 2.53; Na 10.24;

The ninhydrin reaction as well as the TNBS method, which leads to the quantitative and qualitative determination of free amino functions generally used can become negative, d. H. there are no free amino in the polymer functions before. From the percentage sulfur value of the elementary analysis of the The title compound gives a degree of loading of the polymer with 40% β-D-Ga  lactose (equivalent to 33 galactose residues per molecule). The colorimetric Phenol-sulfuric acid determination (Dubois et al., Anal. Chem. 1956, 28, 3, 350) to determine the average degree of pyranose loading of the polyamine provides an average statistical loading of 32.68% of D-galactose residues per molecule and is therefore to be regarded as identical to the degree of loading, which is determined solely by the percentage element ratio se was determined (found here: 33 D-galactose residues per molecule).

c) 13 kD poly-L-lysine (Gd-DTPA amide) ₄₇ - [1- (4-thioureidophenyl) -β-D-galactopyranosyl] ₃₃ conjugate

1.27 g (0.0411 mmol; 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 (1.1 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) as an amorphous powder
Water content: 3.89%

Elemental analysis (calculated on anhydrous substance):
calc .: C 41.00; H 5.45; N 10.11; S 2.31; Gd 16.53; Na 2.42;
Found: C 39.87; H 5.74; N 9.98; S 2.28; Gd 16.58; Na 2.42;

The molar element ratio of gadolinium to sulfur results in thus an average degree of loading of the polymer of 58.9% Gd-DPTA and 41.1 % D-galactose (corresponding to 47 Gd-DTPA units and 33 β-D-galactose residues per molecule). There are no ninhydrin reactions in the title compound free amino functions more detectable.  

Example 2 13 kD poly-L-lysine (Gd-DTPA) ₄₇ - [1- (4-thioureidophenyl) -α- D-mannopyranosyl] ₃₃ conjugate a) 13 kD poly-L-lysine (DTPA amide) ₄₇ - [1- (4-thioureidophenyl) -α-D-mannopyranosyl] ₃₃ conjugate

In an analogous manner to that described for Example 1b), the reaction of 0.75 g (0.023 mmol; corresponding to 0.77 mmol of free amine functions) of the title compound from Example 1a) with 1.2 g (3.8 mmol) of p- Isothiocyanatophenyl-α-D-mannopyranose [Presentation according to: Monsigny, M. et al., Biol. Cell. 51, (1984), 187] after working up 1.29 g (92.4% of theory) of an amorphous and colorless solid.
Water content: 4.86%

Elemental analysis (calculated on anhydrous substance):
calc .: C 44.34; H 5.54; N 11.02; S 2.49; Na 10.18;
Found: C 44.40; H 5.57; N 10.96; S 2.58; Na 10.20;

The ninhydrin reaction on the title compound is negative, d. H. the polymeric product contains no free amino functions. From the pro the central sulfur value of the elementary analysis of the title compound results a degree of loading of the polymer with 40% D-mannose (corresponding to 33 Man residues per molecule).

b) 13 kD-poly-L-lysine (Gd-DTPA amide) ₄₇ - [1- (4-thioureidophenyl) -α- D-mannopyranosyl] ₃₃ conjugate

0.85 g (0.020 mmol; corresponding to 0.94 mmol of DTPA) of the title compound from Example 2a) are described in analogy as for Example 1c), with 10 ml of an aqueous gadolinium chloride solution which has a total content of 0.25 g (0.05 96 mmol) of gadolinium chloride, complexed. After working up and freeze-drying, 918 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 40.39; H 5.05; N 10.04; S 2.27; Gd 15.86; Na 2.32;
Found: C 40.42; H 5.11; N 10.01; S 2.36; Gd 15.92; Na 2.28;

From the molar elementary ratios of gadolinium to sulfur thus an average degree of loading of the polymer of 60.3% Gd-DPTA and 39.7% D-mannose (equivalent to 47 Gd-DTPA units and 33-D-mannose residues per molecule). In the title link are by means of quantitative Ninhy free amino functions no longer detectable in reactions.

The T ₁ and T ₂ relaxivities of the title compound were determined both in water and in plasma:

The measurements were carried out at 40 ° C, a magnetic field strength of 0.47 Tesla, the T1 sequence: 180 ° -TI-90 ° [inversion recovery]
and the T2 sequence: 90 ° -TE-180 ° [CPMG].

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

R ₁ (water) = 14.4 ± 0.0 (L / mmol.s)
R ₁ (plasma) = 17.4 ± 0.2 (L / mmol.s)
R ₂ (water) = 16.4 ± 0.2 (L / mmol.s)
R ₂ (plasma) = 21.4 ± 1.9 (L / mmol.s)

Example 3 13 kD poly-L-lysine (Gd-DTPA amide) ₄₇ - [1- (4-thioureidophenyl) -β-D-glucopy ranosyl] ₃₃ conjugate a) 13 kD poly-L-lysine (DTPA amide) ₄₇ - [1- (4-thioureidophenyl) -β-D-glucopyranosyl] ₃₃ conjugate

In an analogous manner as described for Example 1b), the reaction of 0.9 g (0.028 mmol; corresponding to 0.92 mmol of free amine functions) of the titanium compound from Example 1a) with 1.44 g (4.56 mmol) p -Isothiocyanatophenyl-β-D-glucopyranose [Representation in analogy to: M. Ponpipom et al., J. Med. Chem. 24, 1388; 1981 and Monsigny et al., Bio. Cell, 51, 187, 1984] after working up to 1.08 g (90.6% of theory) of an amorphous and colorless solid.
Water content: 3.75%

Elemental analysis (calculated on anhydrous substance):
calc .: C 44.34; H 5.54; N 11.02; S 2.49; Na 10.18;
Found: C 44.40; H 5.51; N 11.03; S 2.55; Na 10.20;

The ninhydrin reaction of the title compound is negative, i. H. the polymer Product no longer contains free amino functions. From the percentage Sulfur value of the elemental analysis of the title compound gives a Bela Degree of the polymer with 40% D-glucose (corresponding to 33 glucose residues per molecule).

b) 13 kD poly-L-lysine (Gd-DTPA amide) ₄₇ - [1- (4-thioureidophenyl) -β-D-glucopyranosyl] ₃₃ conjugate

1.0 g (0.023 mmol; corresponding to 1.10 mmol of DTPA) of the title compound from Example 3a) are described in analogy as for Example 1c), with 10 ml of an aqueous gadolinium chloride solution which has a total content of 0.298 g (1.12 mmol) has gadolinium chloride solution, complexed. After working up and freeze-drying, 1.06 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 40.39; H 5.05; N 10.04; S 2.27; Gd 15.86; Na 2.32;
Found: C 40.44; H 5.12; N 10.09; S 2.31; Gd 15.90; Na 2.31;

The molar element ratio of gadolinium to sulfur results in thus an average degree of loading of the polymer of 59.7% Gd-DPTA and 40.3 % D-glucose (corresponding to 47 Gd-DTPA units and 33-D-glucose residues per Molecule). There are no free ones in the title compound by means of a ninhydrin reaction Amino functions more detectable.  

Example 4 24-cascade amine (Gd-DTPA amide) ₁₁ - [1- (4-thioureidophenyl) -β-D-galactopyranosyl] ₁₃ conjugate a) 24-cascade amine (DTPA amide) ₁₁ conjugate

At room temperature, 5.0 g (1.06 mmol; corresponding to 25.44 mmol of free amine functions) 24-cascade amine [as HBr salt] are dissolved in 300 ml of distilled water. The pH of the resulting solution is 1.6. A pH of 9.5 is set by dropwise adding this solution with 1 molar sodium hydroxide solution. Then a total of 12.3 g (30.53 mmol) of DTPA monoanhydride monoethyl ester (preparation as described in European Patent Application No. 0263051 A1, European Patent Application No. 0450742 A1 and European Patent Application No. 0413405 A1) are added in portions, wherein the pH of the reaction solution is kept constant at 9.5 by adding 1 molar sodium hydroxide solution. After a reaction time of 20 minutes at room temperature, the pH of the reaction solution is adjusted to 11.5 by adding 1 molar sodium hydroxide solution. The reaction solution is stirred for 12 hours at room temperature and made up to a total volume of 800 ml by adding 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: 5.50 g (86.4 based on the polyamine used) as an amorphous powder
Water content: 9.33%

Elemental analysis (calculated on anhydrous substance):
calc .: C 43.20; H 5.66; N 13.88; Na 12.86;
Found: C 43.01; H 5.70; N 14.03; Na 12.84;

After elemental analysis as well as colorimetric titration with 0.0025 molar Gadolinium sulfate standard solution in the presence of xylenol orange as an indicator, he there is a DTPA degree of loading of the polymer of 45.8% (corresponding to 11  DTPA units per molecule). After quantitative ninhydrin reaction are in Average of 13 free amino functions per polymer molecule.

b) 24-cascade amine (DTPA amide) ₁₁ - [1- (4-thioureidophenyl) -β-D-galactopyranosyl] ₁₃ conjugate

In an analogous manner as described for Example 1b), the reaction of 1.0 g (0.12 mmol; corresponding to 1.65 mmol of free amine functions) of the title compound from Example 4a) with 2.3 g (7.26 mmol) ) p-isothiocyanatophenyl-β-D-galactopyranose after working up 1.33 g (93.2% of theory) of an amorphous and colorless solid.
Water content: 7.41%

Elemental analysis (calculated on anhydrous substance):
calc .: C 45.46; H 5.38; N 10.67; S 3.49; Na 8.47;
Found: C 45.32; H 5.28; N 10.71; S 3.44; Na 8.51;

The ninhydrin reaction title compound is negative, i. H. the polymeric pro product contains no free amino functions. From the percentage sulfur value the elemental analysis of the title compound reveals a degree of loading of the Polymers with 54.2% D-galactose (corresponding to 13 galactose residues per mole cool).

c) 24-cascade amine (Gd-DTPA amide) ₁₁ - [1- (4-thioureidophenyl) -β-D-galactopyranosyl] ₁₃ conjugate

1.0 g (0.077 mmol; corresponding to 0.85 mmol of DTPA) of the title compound from Example 4b) are analogous to that described for Example 1c) with 10 ml of an aqueous gadolinium chloride solution which has a total content of 0.232 g (0.88 mmol) Contains gadolinium chloride, complexed. After working up and freeze-drying, 978.4 mg (98.4% of theory) of the title compound are obtained as amorphous powder.
Water content: 4.36%

Elemental analysis (calculated on anhydrous substance):
calc .: C 42.04; H 4.97; N 9.87; S 3.23; Gd 13.40; Na 1.96;
Found: C 42.12; H 5.03; N 9.81; S 3.26; Gd 13.54; Na 1.93;

The molar element ratio of gadolinium to sulfur results in thus an average degree of loading of the polymer of 46.4% Gd-DPTA and 53.6 % D-galactose (corresponding to 11 Gd-DTPA units and 13-D-galactose residues per molecule). There are no ninhydrin reactions in the title compound free amino functions more detectable.

Example 5 24-cascade amine (Gd-DTPA amide) ₁₁ - [1- (4-thioureidophenyl) -α- D-mannopyranosyl] ₁₃ conjugate a) 24-cascade amine (DTPA amide) ₁₁ - [1- (4-thioureidophenyl) -α- D-mannopyranosyl] ₁₃ conjugate

In an analogous manner as described for Example 1b), the reaction of 1.0 g (0.12 mmol; corresponding to 1.65 mmol of free amine functions) of the title compound from Example 4a) with 2.3 g (7.26 mmol) ) p-isothiocyanatophenyl-α-D-mannopyranose after working up 1.31 g (91.8% of theory) of an amorphous and colorless solid.
Water content: 4.89%

Elemental analysis (calculated on anhydrous substance):
calc .: C 45.46; H 5.38; N 10.67; S 3.49; Na 8.47;
Found: C 45.27; H 5.40; N 10.58; S 3.40; Na 8.60;

The ninhydrin reaction of the title compound is negative, i. H. the polymer Product does not contain any free amino functions. From the percentage sulfur The elemental analysis of the title compound gives a degree of loading  of the polymer with 53.8% D-mannose (corresponding to 20 α-D-mannose units per molecule).

b) 24-cascade amine (Gd-DTPA amide) ₁₁ - [1- (4-thioureidophenyl) -α- D-mannopyranosyl] ₃₃ conjugate

1.0 g (0.077 mmol; corresponding to 0.85 mmol of DTPA) of the title compound from Example 5a) are described in analogy as for Example 1c) with 10 ml of an aqueous gadolinium chloride solution which has a total content of 0.232 g (0.88 mmol ) Contains gadolinium chloride, complexed. After working up and freeze-drying, 952.2 mg (96.4% of theory) of the title compound are obtained as an amorphous powder.
Water content: 7.89%

Elemental analysis (calculated on anhydrous substance):
calc .: C 42.04; H 4.97; N 9.87; S 3.23; Gd 13.40; Na 1.96;
Found: C 41.98; H 5.02; N 10.01; S 3.27; Gd 13.42; Na 1.91;

The molar element ratio of gadolinium to sulfur results in an average degree of loading of the polymer of 45% Gd-DPTA and 55% D- Mannose (corresponding to 11 Gd-DTPA units and 13-D-mannose residues per Molecule). There are no free ones in the title compound by means of a ninhydrin reaction Amino functions more detectable.

The T ₁ and T ₂ relaxivities of the title compound were determined both in water and in plasma:

The measurements were carried out at 40 ° C, a magnetic field strength of 0.47 Tesla and the T ₁ sequence: 180 ° -TI-90 ° [inversion recovery]
and the T ₂ sequence: 90 ° -TE-180 ° [CPMG].

All relaxivity determinations were carried out on a Minispec PC 20. In all cases, a solution of Gd-DTPA with a concentration of 1 mmol / L was used as the reference sample.

R ₁ (water) = 15.4 ± 0.0 (L / mmol.s)
R ₁ (plasma) = 17.7 ± 1.1 (L / mmol.s)
R ₂ (water) = 17.6 ± 0.2 (L / mmol.s)
R ₂ (plasma) = 19.3 ± 1.2 (L / mmol.s)

Example 6 24-cascade amine (Gd-DTPA amide) ₁₁ - [1- (4-thioureidophenyl) -N-acetyl-β-D-glucosaminopyranosyl] ₁₃ conjugate a) 24-cascade amine (DTPA amide) ₁₁ - [1- (4-thioureidophenyl) -N-acetyl β-D-glucosaminopyranosyl] ₁₃ conjugate

In an analogous manner to that described for Example 1b), the reaction of 1.3 g (0.156 mmol; corresponding to 2.03 mmol of free amine functions) of the title compound from Example 4a) with 3.15 g (8.92 mmol) of 2- [Acetylamino-2-deoxy] -1- [4- (isothiocyanatophenyl)] - β-D-glucosaminopyranose [Representation in analogy to: M. Ponpipom et al., J. Med. Chem 24, 1388; 1981 and Monsigny et al., Biol. Cell, 51, 187, 1984] after working up 1.74 g (89.6% of theory) of an amorphous and colorless solid.
Water content: 3.94%

Elemental analysis (calculated on anhydrous substance):
calc .: C 45.97; H 5.56; N 11.66; S 3.34; Na 8.10;
Found: C 46.03; H 5.60; N 11.62; S 3.30; Na 8.07;

The ninhydrin reaction of the title compound is negative, i. H. the polymer Product does not contain any free amino functions. From the percentage sulfur The elemental analysis of the title compound gives a degree of loading of the polymer of 54.1% N-acetyl-glucose units (corresponding to 13 N- Acetylglucosamine residues per molecule).

b) 24-cascade amine (DTPA amide) ₁₁ - [1- (4-thioureidophenyl) -N-acetyl-β-D-glucosaminopyranosyl] ₁₃ conjugate

1.5 g (0.120 mmol; corresponding to 1.32 mmol of DTPA) of the title compound from Example 6a) are complexed in analogy as described for Example 1c) with 10 ml of an aqueous gadolinium chloride solution which has a total content of 0.356 g (1.35 mmol ) Contains gadolinium chloride. After working up and freeze-drying, 1.57 g (97.3% of theory) of the title compound are obtained as an amorphous powder.
Water content: 4.91%

Elemental analysis (calculated on anhydrous substance):
calc .: C 42.65; H 5.16; N 10.82; S 3.10; Gd 12.85; Na 1.88;
Found: C 42.71; H 5.20; N 10.86; S 3.14; Gd 12.91; Na 1.85;

The molar element ratio of gadolinium to sulfur results in an average degree of loading of the polymer of 46% Gd-DTPA and 54% N- Acetyl-D-glucosamine, d. H. there are on average 11 Gd-DTPA units and 13- N-acetyl-D-glucosamine units per molecule. In the title link are no free amino functions can be detected using the ninhydrin reaction.

Example 7 24-cascade amine (Gd-DTPA amide) ₁₁ - [1- (4-thioureidophenyl) -β-D-glucopyranosyl] ₁₃ conjugate a) 24-cascade amine (DTPA amide) ₁₁ - [1- (4-thioureidophenyl) -β-D-glucopyranosyl] ₁₃ conjugate

In an analogous manner as described for Example 1b), the reaction of 1.2 g (0.144 mmol; corresponding to 1.872 mmol of free amine functions) of the title compound from Example 4a) with 2.61 g (8.24 mmol) of p-isothiocyanatophenyl β -D-glucopyranose after working up 1.49 g (87.1% of theory) of an amorphous and colorless solid.
Water content: 8.37%

Elemental analysis (calculated on anhydrous substance):
calc .: C 45.46; H 5.38; N 10.67; S 3.49; Na 8.47;
Found: C 45.52; H 5.44; N 10.73; S 3.55; Na 8.60;

The ninhydrin reaction of the title compound is negative, i. H. the polymer Product does not contain any free amino functions. From the percentage sulfur  The elemental analysis of the title compound gives a degree of loading of the polymer with 54.9% D-glucose (corresponding to 13 glucose residues per mole cool).

b) 24-cascade amine (Gd-DTPA amide) ₁₁ - [1- (4-thioureidophenyl) -β-D-glucopyranosyl] ₁₃ conjugate

1.0 g (0.077 mmol; corresponding to 0.85 mmol of DTPA) of the title compound from Example 7a) are complexed with 232 mg (0.88 mmol) of an aqueous gadolinium chloride solution (10 ml) in a manner analogous to that described for Example 1c). After working up and freeze-drying, 952.2 mg (96.4% of theory) of the title compound are obtained as an amorphous powder.
Water content: 7.89%

Elemental analysis (calculated on anhydrous substance):
calc .: C 42.04; H 4.97; N 9.87; S 3.23; Gd 13.40; Na 1.96;
Found: C 41.98; H 5.02; N 10.01; S 3.27; Gd 13.42; Na 1.91;

The molar element ratio of gadolinium to sulfur results in an average degree of loading of the polymer of 45% Gd-DPTA and 55% D- Glucose (corresponding to 11 Gd-DTPA units and 13-D-glucose residues per Molecule). There are no free ones in the title compound by means of a ninhydrin reaction Amino functions more detectable.

Example 8 36-cascade amine (Gd-DTPA amide) ₁₆ - [1- (4-thioureidophenyl) -β-D-galactopyranosyl] ₂₀ conjugate a) 36-cascade amine (DTPA amide) ₁₆ conjugate

At room temperature, 4.0 g (0.54 mmol; corresponding to 19.41 mmol of free amine functions) 36-cascade amine [used as hydrobromide; Representation according to: Schmitt-Willich et al., Schering AG, European Patent Application No. 0430 863 A1; WO 97/02051; German patent application No. 195 18 222 A1] dissolved in 300 ml of distilled water. The pH of the resulting solution is 1.8. A pH of 9.5 is set by dropwise adding this solution with 1 molar sodium hydroxide solution. Then a total of 9.38 g (23.3 mmol) of DTPA monoanhydride monoethyl ester are added in portions, the pH of the reaction solution being kept constant at 9.5 by adding 1 molar sodium hydroxide solution. After a reaction time of 20 minutes at room temperature, the pH of the reaction solution is adjusted to 11.5 by setting with 1 molar sodium hydroxide solution. The reaction solution is stirred for 12 hours at room temperature and made up to a total volume of 800 ml by adding 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: 5.50 g (86.4% of theory; based on the polyamine used) as an amorphous powder
Water content: 9.33%

Elemental analysis (calculated on anhydrous substance):
calc .: C 43.23; H 5.63; N 13.88; Na 12.46;
Found: C 43.40; H 5.68; N 13.92; Na 12.30;

After elemental analysis as well as colorimetric titration with 0.0025 molar Gadolinium sulfate standard solution in the presence of xylenol orange as an indicator, he there is a DTPA loading level of the polymer of 45.1% (corresponding to 16 DTPA units per molecule). After the ninhydrin reaction of the title compound gene on average in the polymer 20 free amino functions per molecule.

b) 36-cascade amine (DTPA amide) ₁₆ - [1- (4-thioureidophenyl) -β-D-galactopyranosyl] ₂₀ conjugate

In an analogous manner as described for Example 1b), the reaction of 1.5 g (0.12 mmol; corresponding to 2.54 mmol of free amine functions) of the title compound from Example 8a) with 3.55 g (11.2 mmol) ) p-isothiocyanatophenyl-α-D-mannopyranose after working up 1.87 g (86.3% of theory) of an amorphous and colorless solid.
Water content: 4.93%

Elemental analysis (calculated on anhydrous substance):
calc .: C 45.52; H 5.35; N 10.62; S 3.55; Na 8.14;
Found: C 45.60; H 5.41; N 10.58; S 3.49; Na 8.22;

The ninhydrin reaction of the title compound is negative, i. H. the polymer Product does not contain any free amino functions. From the percentage sulfur The elemental analysis of the title compound gives a degree of loading of the polymer with 59% N-acetyl-D-glucosamine (corresponding to 20 glucose residues per molecule).

c) 36-cascade amine (Gd-DTPA amide) ₁₆ - [1- (4-thioureidophenyl) -β-D-galactopyranosyl] ₂₀ conjugate

1.5 g (0.10 mmol; corresponding to 1.32 mmol of DTPA) of the title compound from Example 8b) are described in analogy as for Example 1c) with a solution of 358.5 mg (1.36 mmol) gadolinium chloride in 10 ml of water. After working up and freeze-drying, 1.57 g (97.3% of theory) of the title compound are obtained as an amorphous powder.
Water content: 7.71%

Elemental analysis (calculated on anhydrous substance):
calc .: C 42.22; H 4.96; N 9.85; S 3.29; Gd 12.91; Na 1.89;
Found: C 42.30; H 5.02; N 9.78; S 3.31; Gd 13.01; Na 1.78;

The molar element ratio of gadolinium to sulfur results in thus an average degree of loading of the polymer of 44% Gd-DPTA and 56% D-mannose (corresponding to 16 Gd-DTPA units and 20 D-mannose residues per molecule). There are no free ones in the title compound by means of a ninhydrin reaction Amino functions more detectable.  

Example 9 36-cascade amine (Gd-DTPA amide) ₁₆ - [1- (4-thioureidophenyl) -β-D-glucopy ranosyl] ₂₀ conjugate a) 36-cascade amine (DTPA amide) ₁₆ - [1- (4-thioureidophenyl) -β-D-glucopyranosyl] ₂₀ conjugate

In an analogous manner as described for Example 1b), the reaction of 2.5 g (0.20 mmol; corresponding to 4.0 mmol of free amine functions) of the title compound from Example 8a) with 5.58 g (17.6 mmol) ) p-isothiocyanatophenyl-β-D-glucopyranose after working up 3.26 g (90.2% of theory) of an amorphous and colorless solid.
Water content: 8.44%

Elemental analysis (calculated on anhydrous substance):
calc .: C 45.52; H 5.35; N 10.62; S 3.55; Na 8.14;
Found: C 45.61; H 5.42; N 10.57; S 3.49; Na 8.13;

The ninhydrin color reaction test for the presence of free amino radio ions in the title link is negative, i. H. contains the polymeric product thus no more free amino functions. From the percentage sulfur value the elemental analysis of the title compound reveals a degree of loading of the Polymers with 56.2% D-glucose (corresponding to 20 glucose residues per molecule).

b) 36-cascade amine (Gd-DTPA amide) ₁₆ - [1- (4-thioureidophenyl) -β-D-glucopyranosyl] ₂₀ conjugate

3.0 g (0.166 mmol; corresponding to 2.65 mmol of DTPA) of the title compound from Example 9a) are complexed in a manner analogous to that described for Example 1c) with a solution of 717 mg (2.72 mmol) of gadolinium chloride in 15 ml of water. After working up and freeze-drying, 3.18 g (98.4% of theory) of the title compound are obtained as an amorphous powder.
Water content: 6.43%

Elemental analysis (calculated on anhydrous substance):
calc .: C 42.22; H 4.96; N 9.85; S 3.29; Gd 12.91; Na 1.89;
Found: C 42.24; H 4.93; N 10.01; S 3.27; Gd 12.99; Na 1.91;

The molar element ratio of gadolinium to sulfur results in an average degree of loading of the polymer of 44% Gd-DPTA and 56% β-D- Glucose (corresponding to 16 Gd-DTPA units and 20 β-D-glucose residues per Molecule). There are no free ones in the title compound by means of a ninhydrin reaction Amino functions more detectable.

Example 10 24-cascade amine (Gd-Gly-Me-DOTA amide) ₁₁ - (α-D-mannopyranosyl-1-thio-3-propionic acid amide) ₁₃ conjugate a) 24-cascade amine (Gly-Me-DOTA-tri-t.-butyl ester amide) ₁₁

20.0 g (26.8 mmol) Gly-Me-DOTA-tri-tert.butyl ester (title compound from Bei game 31f) are at room temperature in 100 ml of absolute dimethylformamide dissolved and mixed with 3.7 g (32.16 mmol) of N-hydroxysuccinimide. The clear reaction mixture is then cooled to 0 ° C. and at this temperature in portions with a total of 6.6 g (32.16 mmol) of dicyclo hexylcarbodiimide added. After a reaction time of 30 minutes at 0 ° C and The precipitated dicyclohexylurea is removed for 2.5 hours at room temperature filtered off. The filtrate thus obtained is now slowly added dropwise at room temperature to a stirred solution of 8.8 g (1.86 mmol; corresponding to 44.8 mmol free amino functions) 24-part cascade polyamine and 4.5 g (44.8 mmol) Triethylamine in 60 ml of absolute DMF. After a response time of 12 hours at room temperature is evaporated to dryness in vacuo and the ver remaining residue with 250 ml of dichloromethane. After filtering off of insoluble parts, the organic phase is successively with 5% Hydrochloric acid, saturated sodium bicarbonate solution and twice with What washed. After drying the organic product phase over Na Triumsulfat is filtered and the solvent removed in vacuo. Man he holds 14.03 g (84.3% of theory; based on the polyamine used) easily yellow colored oil, which without further cleaning immediately for the after following synthesis steps is used. After quantitative ninhydrin freak  tion of the above title compound are on average 13 free amine functions per molecule.

Elemental analysis:
calc .: C 63.06; H 9.68; N 15.65;
Found: C 63.20; H 10.02; N 15.71;

b) 24-cascade amine (Gly-Me-DOTA-tri-t-butyl ester amide) ₁₁ - [3- (2,3,4,6-tetra-O-acetyl-1-thio) -α-D -mannopyranosyl-3-propion-amide] ₁₃ - conjugate

A stirred solution of 3.5 g (0.39 mmol; corresponding to 5.1 mmol of free Amine functions) of the title compound from Example 10a) in 100 ml of absolute Dichloromethane, at room temperature with 617 mg (6.1 mmol) of triethylamine transferred. Then with a solution of 2.97 g (5.61 mmol) of 3- (2,3,4,6- Tetra-O-acetyl-1-thio-α-D-mannopyranosyl) propionic acid N-hydroxysuccinimide ester (representation in analogy to: J. Haensler et al., Bioconjugate Chem. 4, 85, (1993); Chipowsky, S., and Lee, Y. C (1973), Synthesis of 1-Thio-aldosides; Carbohydrate Research 31, 339-346) in 60 ml dichloromethane, at room temperature ratured in drops. After a reaction time of 12 hours at Room temperature is adjusted to 99999 00070 552 001000280000000200012000285919988800040 0002019728954 00004 99880 dichloromethane to a total volume of 500 ml replenished. The organic phase is washed twice with water and washed twice with saturated sodium bicarbonate solution and over Dried sodium sulfate. After filtering, the solvent in Va vacuum stripped to dryness. 5.0 g (89.7% of theory) of the title ver binding as a colorless oil. The ninhydrin reaction of the title compound proceeds negative, d. H. free amino functions are no longer detectable in the polymer.

Elemental analysis:
calc .: C 57.67; H 8.03; N 9.73; S 2.90;
Found: C 57.53; H 7.92; N 9.79; S 2.87;

Hence the percentage sulfur value of the elementary analysis gives the above-mentioned title compound with an average degree of loading of the polymer 55% D-mannose tetraacetate residues (corresponding to 13 D-mannose units per Molecule).  

c) 24-cascade amine (Gly-Me-DOTA amide) ₁₁ - (-α-D-mannopyranosyl-1-thio-3-propione amide) ₁₃ conjugate

4.5 g (0.312 mmol) of the title compound from Example 10b) are dissolved in 100 ml of pure 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 concentrated 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 at room temperature with 50 ml of a 32% aqueous ammonia solution. 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 brought to pH 3.0 by adding 10% hydrochloric acid and 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: 2.80 g (81.6% of theory) as an amorphous and colorless powder
Water content: 7.78%

Elemental analysis (calculated on anhydrous substance):
calc .: C 49.42; H 7.09; N 12.67; S 3.77;
Found: C 49.18; H 7.13; N 12.81; S 3.69;

After elemental analysis as well as colorimetric titration with 0.0025 molar Gadolinium sulfate standard solution, in the presence of xylenol orange as an indicator, the degree of loading with Gly Me-DOTA of the polymer is 44.8% (corresponding to 11 Gly-Me DOTA units per molecule). After Ninhydrinreak tion there are no free amino functions in the polymer.

d) 24-cascade amine (Gd-Gly-Me-DOTA amide) ₁₁ - (α-D-mannopyranosyl-1-thio-3-propionic acid amide) ₁₃ conjugate

2.5 g (0.22 mmol; corresponding to 2.48 mmol of Gly-Me-DOTA) of the title compound solution from Example 10c) are dissolved in 70 ml of distilled water and the pH  The value of the solution is determined by dropwise addition with 10% aqueous salt acidity set to 2.0. Finally, 670 mg are added at room temperature (2.55 mmol) gadolinium chloride dissolved in 10 ml water and stirred the Re Action solution for 12 hours at 60 ° C. At room temperature the reak tion solution brought to pH 7.2 by adding 1 molar sodium hydroxide solution made up to a total volume of 500 ml with distilled water. After three times ultrafiltration against distilled water via a YM3 ultrafiltra tion membrane (AMICON®), the remaining residue is freeze-dried net. 2.7 g (97.3% of theory) of the title compound are obtained as an amorphous, colorless Powder with a water content of 9.37%.

Elemental analysis (calculated on anhydrous substance):
calc .: C 42.77; H 6.05; N 19.96; S 3.26; Gd 13.54;
Found: C 42.69; H 6.18; N 11.04; S 3.38; Gd 13.67;

From the molar element ratios of sulfur to gadolinium (1.21) he there is thus an average degree of loading of the polymer of 45.2% Gd-Gly- Me-DOTA and 54.8% α-D-mannose (corresponding to 11 Gd-Gly-Me-DOTA-Ein units and 13-α-D-mannose residues per molecule). In the title link are no free amino functions can be detected using the ninhydrin reaction.

Example 11 24-cascade amine (Gd-Gly-Me-DOTA amide) ₁₁ - (β-D-glucopyranosyl-1-thio-3-propionic acid amide) ₁₃ conjugate a) 24-cascade amine (Gly-Me-DOTA-tri-t-butyl ester amide) ₁₁ - [3- (2,3,4,6-tetra-O-acetyl-1-thio) -β-D -glucopyranosyl-3-propion-amide] ₁₃ - conjugate

In an analogous manner as described for Example 10b), the implementation of 3.0 g (0.33 mmol; corresponding to 4.3 mmol of free amine functions) of the title Compound from Example 10a) with 2.76 g (5.2 mmol) of 3- (2,3,4,6-tetra-O-acetyl-1- thio-β-D-glucopyranosyl) propionic acid N-hydroxysuccinimide ester, 4.0 g (85.3 % d. Th.) Of the title compound as a colorless oil.  

Elemental analysis:
calc .: C 57.67; H 8.03; N 9.73; S 2.90;
Found: C 57.39; H 8.14; N 9.87; S 2.96;

The title compound shows a negative ninhydrin reaction. From the percent len sulfur value of the elementary analysis gives an average degree of loading of Polymers with 13 D-glucose residues per molecule.

b) 24-cascade amine (Gly-Me-DOTA amide) ₁₁ - (β-D-glucopyranosyl-1-thio-3-propionic acid amide) ₁₃ conjugate

In an analogous manner to that described for Example 10c), the deprotection of 3.5 g (0.24 mmol) of the title compound from Example 11a) leads to the formation of 2.2 g (83.6% of theory) after freeze drying. ) the above-mentioned title compound as an amorphous powder.
Water content: 4.73%

Elemental analysis (calculated on anhydrous substance):
calc .: C 49.42; H 7.09; N 12.67; S 3.77;
Found: C 50.01; H 7.14; N 12.62; S 3.81;

c) 24-cascade amine (Gd-Gly-Me-DOTA amide) ₁₁ - (β-D-glucopyranosyl-1-thio-3-propionic acid amide) ₁₃ conjugate

In an analogous manner as described for Example 10d), the complexation of 2.0 g (0.176 mmol; corresponding to 1.94 mmol of Gly-Me-DOTA) of the title compound from Example 11b) with 520 mg (1.98 mmol) Gadolinium chloride dissolved in 10 ml of water to form the title compound mentioned above.
Yield: 2.75 g (98.1% of theory) as a colorless and amorphous powder
Water content: 9.36%

Elemental analysis (calculated on anhydrous substance):
calc .: C 42.77; H 6.05; N 10.96; S 3.26; Gd 13.54;
Found: C 42.73; H 6.14; N 11.06; S 3.30; Gd 13.61;

Average degree of loading per molecule of the above title compound:
11 Gd-Gly-Me-DOTA units
and 13 D-glucopyranosyl residues

Example 12 24-cascade amine (Gd-Gly-Me-DOTA amide) ₁₁ - (β-D-galactopyranosyl-1-thio-3-propionic acid amide) ₁₃ conjugate a) 24-cascade amine (Gly-Me-DOTA-tri-t-butyl ester amide) ₁₁ - [3- (2,3,4,6-tetra-O-acetyl-1-thio) -β-D -galactopyranosyl-3-propion-amide] ₁₃ - conjugate

In an analogous manner as described for Example 10b), the implementation of 1.5 g (0.165 mmol; corresponding to 2.14 mmol of free amine functions) Title compound from Example 10a) with 1.36 g (2.57 mmol) of 3- (2,3,4,6-tetra-O- acetyl-1-thio-β-D-galactopyranosyl) propionic acid N-hydroxysuccinimide ester 2.0 g (87.2% of theory) of the title compound as a colorless oil.

Elemental analysis:
calc .: C 57.67; H 8.03; N 9.73; S 2.90;
Found: C 57.78; H 8.14; N 9.80; S 2.97;

The title compound shows a negative ninhydrin reaction. From the percent len sulfur value of the elementary analysis results in an average degree of loading of the polymer with 13 D-galactose residues per molecule.

b) 24-cascade amine (Gly-Me-DOTA amide) ₁₁ - (β-D-galactopyranosyl-1-thio-3-propionic acid amide) ₁₃ conjugate

In an analogous manner to that described for Example 10c), the deprotection of 1.8 g (0.12 mmol) of the title compound from Example 12a) leads to the formation of 1.15 g (87.2% of theory) after freeze drying. ) the above-mentioned title compound as an amorphous powder.
Water content: 4.22%

Elemental analysis (calculated on anhydrous substance):
calc .: C 49.42; H 7.09; N 12.67; S 3.77;
Found: C 49.66; H 7.13; N 12.54; S 3.70;

c) 24-cascade amine (Gd-Gly-Me-DOTA amide) ₁₁ - (β-D-galactopyranosyl-1-thio-3-propionic acid amide) ₁₃ conjugate

In an analogous manner to that described for Example 10d), the complexation of 1.0 g (0.088 mmol; corresponding to 1.94 mmol of Gly-Me-DOTA) of the title compound from Example 12b) with 262 mg (1.0 mmol) Gadolinium chloride dissolved in 10 ml of water to form the title compound mentioned above.
Yield: 1.08 g (96.4% of theory) as a colorless and amorphous powder
Water content: 9.32%

Elemental analysis (calculated on anhydrous substance):
calc .: C 42.77; H 6.05; N 10.96; S 3.26; Gd 13.54;
Found: C 42.56; H 6.12; N 10.93; S 3.24; Gd 13.51;

Average degree of loading per molecule of the above title compound:
11 Gd-Gly-Me-DOTA units
and 13 D-galactopyranosyl residues

Example 13 24-cascade amine (Gd-Gly-Me-DOTA amide) ₁₁ - (N-acetyl-β-D-glucosamino pyranosyl-1-thio-3-propionic acid amide) ₁₃ conjugate a) 24-cascade amine (Gly-Me-DOTA-tri-t-butyl ester amide) ₁₁ - [3- (N-acetyl-3,4,6-tetra-O-acetyl-1-thio) -β -D-glucosaminopyranosyl-3-propione-amide] ₁₃ conjugate

In an analogous manner as described for Example 10b), the implementation of 2.0 g (0.22 mmol; corresponding to 2.86 mmol of free amine functions) the title Compound from Example 10a) with 1.74 g (3.43 mmol) 4 - [(Acetylamino-4-deoxy)] - 3- (2,3,4,6-tetra-O-acetyl-1-thio-β-D-glucosaminopyranosyl) propionic acid-N-hy droxysuccinimide ester 2.6 g (83.5% of theory) of the title compound as a colorless oil.  

Elemental analysis:
calc .: C 57.77; H 8.04; N 11.02; S 2.90;
Found: C 57.68; H 8.12; N 11.13; S 2, 87;

The title compound shows a negative ninhydrin reaction. From the percent len sulfur value of the elementary analysis results in an average degree of pollution of the polymer with tetraacetyl-glucosamine residues per molecule.

b) 24-cascade amine (Gly-Me-DOTA amide) ₁₁ - (N-acetyl-β-D-glucosaminopyranosyl-1-thio-3-propionic acid amide) ₁₃ conjugate

In an analogous manner as described for Example 10c), the deprotection of 2.5 g (0.17 mmol) of the title compound from Example 13a) leads to the formation of 1.74 g (88.3% of theory) after freeze drying. ) the above-mentioned title compound as an amorphous powder.
Water content: 10.02%

Elemental analysis (calculated on anhydrous substance):
calc .: C 49.84; H 7.10; N 13.65; S 3.60;
Found: C 50.02; H 7.18; N 13.72; S 3.67;

c) 24-cascade amine (Gd-Gly-Me-DOTA amide) ₁₁ - (N-acetyl-β-D-glucosaminopyranosyl-1-thio-3-propionic acid amide) ₁₃ conjugate

In an analogous manner as described for Example 10d), the complexation of 1.5 g (0.13 mmol; corresponding to 1.42 mmol of Gly-Me-DOTA) of the title compound from Example 13b) leads to 380 mg (1.45 mmol) gadolinium chloride, dissolved in 10 ml of water, to form the above-mentioned title compound.
Yield: 1.69 g (98.0% of theory) as a colorless and amorphous powder
Water content: 7.44%

Elemental analysis (calculated on anhydrous substance):
calc .: C 43.48; H 5.95; N 11.91; S 3.14; Gd 13.02;
Found: C 43.55; H 6.04; N 11.88; S 3.18; Gd 12.98;

Average degree of loading per molecule of the above title compound:
11 Gd-Gly-Me-DOTA units and
13 N-acetyl-D-glucosaminopyranosyl residues

Example 14 48-Starburst dendrimer (Gd-DTPA amide) ₁₉ - [1- (4-thioureidophenyl) -β-D-galactopyranosyl] ₂₉ conjugate a) 48-Starburst-Dendrimer (DTPA amide) ₁₉ conjugate

5.0 g (0.47 mmol; corresponding to 22.6 mmol of free amine functions) 48-star burst cascade amine (Tomalia DA, et al .; Polym. J. (Tokyo) 1985, 17, 117; Tomalia DA et al. ; Macromolecules 1986, 19, 2466; Tomalia DA, et al .; Macromolecules 1987, 20, 1164) are dissolved in 400 ml of distilled water and the pH is adjusted to 9.5 with 1 molar sodium hydroxide solution. Finally, a total of 10.9 g (27.12 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 is ultra-filtered 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: 8.06 g (88.3%; based on the polyamine used) as an amorphous powder
Water content: 7.32%

Elemental analysis (calculated on anhydrous substance):
calc .: C 45.18; H 6.50; N 17.59; Na 8.99;
Found: C 45.26; H 6.61; N 17.32; Na 9.04;

After elemental analysis as well as colorimetric titration with 0.0025 molar Gadolinium sulfate standard solution, in the presence of xylenol orange as an indicator, the polymer has a DTPA loading level of 40.1% (correspondingly 19 DTPA units per molecule). After quantitative ninhydrin reaction [Lit .: Chem. Rev., 60, 39 (1960); Tetrahedron, 34, 1285 (1978); 47, 8791 (1991)] lie gene on average in the polyamine 29 free amino functions per molecule.

b) 48-Starburst dendrimer (DTPA amide) ₁₉ - [1- (4-thioureidophenyl) β-D-galactopyranosyl] ₂₉ conjugate

1.0 g (0.051 mmol; corresponding to 1.5 mmol of free amino functions) of the titanium compound from Example 14a) are dissolved in 80 ml of deionized water and the pH of the solution is adjusted to 9.5 with 1 molar hydrochloric acid. A total of 1.55 g (4.9 mmol) of p-isothiocyanatophenyl-β-D-galactopyranose is then added in portions, the pH of the reaction solution being kept constant at 9.5 by means of 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 1.34 g (92% of theory) of an amorphous and colorless solid.
Water content: 4.9%

Elemental analysis (calculated on anhydrous substance):
calc .: C 46.62; H 6.06; N 13.39; S 3.26; Na 6.12;
Found: C 46.71; H 6.13; N 13.28; S 3.20; Na 6.21;

The ninhydrin reaction of the title compound is negative, i. H. lie in the polymer no free amino functions. From the percentage of sulfur Elemental analysis of the title compound reveals a degree of loading of the poly mer with 60% D-galactose (corresponding to 29 galactose residues per molecule).  

c) 48-Starburst dendrimer (Gd-DTPA amide) ₁₉ - [1- (4-thioureidophenyl) - β-D-galactopyranosyl] ₂₉ conjugate

1.0 g (6.035 mmol; corresponding to 0.66 mmol of DTPA) of the title compound from Example 14b) are dissolved in 40 ml of sodium citrate buffer (pH 5.3) and at room temperature with a solution of 0.18 g (0.7 mmol) of gadolinium chloride, dissolved 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: 1.0 g (95% of theory) as an amorphous powder
Water content: 5.23%

Elemental analysis (calculated on anhydrous substance):
calc .: C 44.06; H 5.67; N 12.66; S 13.08; Gd 9.89; Na 1.45;
Found: C 44.20; H 5.71; N 12.59; S 13.11; Gd 10.01; Na 1.47;

The molar element ratio of gadolinium to sulfur results in thus an average degree of loading of the polymer of 40% Gd-DPTA and 60% D-galactose (corresponding to 19 Gd-DTPA units and 29-D-galactose residues per molecule). In the title compound, none are free by means of ninhydrin reactions Amino functions more detectable.

Example 15 48-Starburst dendrimer (Gd-DTPA amide) ₁₉ - [1- (4-thioureidophenyl) -α- D-mannopyranosyl] ₂₉ conjugate a) 48-Starburst dendrimer (DTPA amide) ₁₉ - [1- (4-thioureioureidophenyl) -α- D-mannopyranosyl] ₂₉ conjugate

In an analogous manner as described for Example 14b), the reaction of 0.75 g (0.038 mmol; corresponding to 1.12 mmol of free amine functions) of the title compound from Example 14a) with 1.2 g (3.8 mmol) p -Isothiocyanatophenyl-α-D-mannopyranose [representation according to: Monsigny, M. et al., Biol. Cell, 51, 187 (1984)] after working up 0.94 g (89.2% of theory) of an amorphous and colorless solid.
Water content: 5.03%

Elemental analysis (calculated on anhydrous substance):
calc .: C 46.62; H 6.06; N 13.39; S 3.26; Na 6.12;
Found: C 46.70; H 6.01; N 13.42; S 3.24; Na 6.09;

The ninhydrin reaction of the title compound is negative, i. H. the polymer Product does not contain any free amino functions. From the percentage sulfur The elemental analysis of the title compound gives a degree of loading of the polymer with 60% D-mannose (corresponding to 29 mannose residues per mo lekül).

b) 48-Starburst dendrimer (Gd-DTPA amide) ₁₉ - [1- (4-thioureidophenyl) -α- D-mannopyranosyl] ₂₉ conjugate

0.85 g (0.029 mmol; corresponding to 0.56 mmol of DTPA) of the title compound from Example 15a) are described in analogy as described for Example 14c) with 0.15 g (0.58 mmol) of gadolinium chloride, dissolved in 10 ml of distilled water , complex. After working up and freeze-drying, 854 mg (97.5% of theory) of the title compound are obtained as an amorphous powder.
Water content: 6.14%

Elemental analysis (calculated on anhydrous substance):
calc .: C 44.06; H 5.76; N 12.66; S 3.08; Gd 9.89; Na 1.45;
Found: C 44.10; H 5.66; N 12.71; S 3.12; Gd 9.94; Na 1.38;

The molar element ratio of gadolinium to sulfur results in thus an average degree of loading of the polymer of 40% Gd-DPTA and 60% D-mannose (corresponding to 19 Gd-DTPA units and 29-D-mannose residues per molecule). There are no ninhydrin reactions in the title compound free amino functions more detectable.  

Example 16 48-Starburst dendrimer (Gd-DTPA amide) ₁₉ - [1- (4-thioureidophenyl) -β-D-glucopyranosyl] ₂₉ conjugate a) 48-Starburst dendrimer (DTPA amide) ₁₉ - [1- (4-thioureidophenyl) β-D-glucopyranosyl] ₂₉ conjugate

In an analogous manner as described for Example 14b), the reaction of 1.0 g (0.051 mmol; corresponding to 1.5 mmol of free amine functions) of the title compound from Example 14a) with 1.44 g (4.56 mmol) p -Isothiocyanatophenyl-β-D-glucopyranose [purchased goods: Sigma or representation analogous to: Monsigny et al., Biol. Cell., 51, 187 (1984)] after working up 1.36 mg (94% of theory) of one amorphous and colorless solid.
Water content: 5.08%

Elemental analysis (calculated on anhydrous substance):
calc .: C 46.62; H 6.06; N 13.39; S 3.26; Na 6.12;
Found: C 46.55; H 6.12; N 13.29; S 3.24; Na 6.01;

The ninhydrin reaction of the title compound is negative, i. H. the polymer Product does not contain any free amino functions. From the percentage sulfur The elemental analysis of the title compound gives a degree of loading of the polymer with 60% D-glucose (corresponding to 29 glucose residues per mole cool).

b) 48-Starburst dendrimer (Gd-DTPA amide) ₁₉ - [1- (4-thioureidophenyl) -β-D-glucopyranosyl] ₂₉ conjugate

1.0 g (0.034 mmol; corresponding to 0.646 mmol of DTPA) of the title compound from Example 16a) are analogous to that described for Example 14c) with 0.17 g (0.66 mmol) dissolved in 10 ml of distilled water, a gadolinium chloride solution complexed. After working up and freeze-drying, 998 mg (97.2% of theory) of the title compound are obtained as an amorphous powder.
Water content: 3.94%

Elemental analysis (calculated on anhydrous substance):
calc .: C 44.06; H 5.67; N 12.66; S 3.08; Gd 9.89; Na 1.45;
Found: C 44.12; H 5.67; N 12.74; S 3.00; Gd 10.01; Na 1.38;

The molar element ratio of gadolinium to sulfur results in thus an average degree of loading of the polymer of 40% Gd-DPTA and 60% D-glucose (corresponding to 19 Gd-DTPA units and 29-D-glucose residues per Molecule). In the title compound, none are free by means of the ninhydrin reactions Amino functions more detectable.

Example 17 48-Starburst dendrimer (Gd-DTPA amide) ₁₉ - [1- (4-thioureidophenyl) -α-L-fucopyranosyl] ₂₉ conjugate a) 48-Starburst dendrimer (DTPA amide) ₁₉ - [1- (4-thioureidophenyl) -α- L-fucopyranosyl] ₂₉ conjugate

In an analogous manner as described for Example 14b), the reaction of 1.0 g (0.051 mmol; corresponding to 1.48 mmol of free amine functions) of the titanium compound from Example 14a) with 1.65 g (5.57 mmol) 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 1.36 g (94% of theory) of an amorphous and colorless solid.
Water content: 7.24%

Elemental analysis (calculated on anhydrous substance):
calc .: C 47.39; H 6.16; N 13.62; S 3.31; Na 6.22;
Found: C 47.42; H 6.08; N 13.57; S 2.98; Na 6.30;

The ninhydrin reaction of the title compound is negative, i. H. the polymer Product does not contain any free amino functions. From the percentage sulfur The elemental analysis of the title compound gives a degree of loading of the polymer with 60% L-fucose (corresponding to 29 α-L-fucose residues per mo lekül).  

b) 48-Starburst dendrimer (Gd-DTPA amide) ₁₉ - [1- (4-thioureidophenyl) - α-L-fucopyranosyl] ₂₉ conjugate

1.2 g (0.042 mmol; corresponding to 0.81 mmol of DTPA) of the title compound from Example 17a) are analogous to that described for Example 14c) with 0.21 g (0.83 mmol) of an aqueous gadolinium chloride solution, dissolved in 10 ml distilled water, complexed. After working up and freeze-drying, 1.25 g (98.5% of theory) of the title compound are obtained as an amorphous powder.
Water content: 4.39%

Elemental analysis (calculated on anhydrous substance):
calc .: C 44.72; H 5.81; N 12.89; S 3.12; Gd 10.04; Na 1.47;
Found: C 44.81; H 5.79; N 12.78; S 3.16; Gd 10.09; Na 1.40;

The molar element ratio of gadolinium to sulfur results in thus an average degree of loading of the polymer of 40% Gd-DPTA and 60% α-L-fucose (corresponding to 19 Gd-DTPA units and 29 α-L-fucose residues per molecule). In the title compound, none are free by means of ninhydrin reactions Amino functions more detectable.

Example 18 24-Starburst dendrimer (Gd-DTPA amide) ₁₀ - [1- (4-thioureidophenyl) -β-D-galactopyranosyl] ₁₄ conjugate a) 24-Starburst dendrimer (DTPA amide) ₁₀ conjugate

A solution of 5.0 g (0.97 mmol, corresponding to 23.3 mmol of free amine functions) 24-star burst dendrimer polyamine (representation according to: Tomalia DA et al .; Macromolecules 1986, ii, 2466; Tomalia DA et al .; Macromo lecules 1987, 20, 1164) is dissolved in 400 ml of distilled water and adjusted to a pH of 9.5 with 1 molar sodium hydroxide solution. A total of 11.2 g (27.9 mmol) of DTPA monoanhydride monoethyl ester are then 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 YMB 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: 4.6 g (92% of theory; based on the polyamine used) as an amorphous powder
Water content: 8.37%

Elemental analysis (calculated on anhydrous substance):
calc .: C 44.72; H 6.58; N, 17.29; Na 9.38;
Found: C 44.64; H 6.51; N 17.36; Na 9.42;

After elemental analysis as well as colorimetric titration with 0.0025 molar Gadolinium sulfate standard solution in the presence of xylenol orange as an indicator, he there is a DTPA loading level of the polymer of 41.6% (corresponding to 10 DTPA units per molecule). After quantitative ninhydrin reaction are in Means 14 free amino functions per molecule in the polymer.

b) 24-Starburst dendrimer (DTPA amide) ₁₀ - [1- (4-thioureidophenyl) β-D-galactopyranosyl] ₁₄ conjugate

In an analogous manner as described for Example 1b), the reaction of 1.0 g (0.10 mmol; corresponding to 1.43 mmol of free amine functions) of the title compound from Example 18a) with 1.60 g (5.03 mmol) ) p-isothiocyanatophenyl-β-D-galactopyranose after working up 1.32 g (93.3% of theory) of an amorphous and colorless solid.
Water content: 7.31%

Elemental analysis (calculated on anhydrous substance):
calc .: C 46.25; H 6.13; N 13.31; S 3.16; Na 6.47;
Found: C 46.32; H 6.19; N 13.28; S 3.09; Na 6.51;

The ninhydrin reaction of the title compound is negative, i. H. the polymer Product does not contain any free amino functions. From the percentage sulfur  The elemental analysis of the title compound gives a degree of loading of the polymer with 59% D-galactose (corresponding to 14 galactose residues per Molecule).

c) 24-Starburst dendrimer (Gd-DTPA amide) ₁₀ - [1- (4-thioureidophenyl) -β-D-galactopyranosyl] ₁₄ conjugate

1.0 g (0.07 mmol; corresponding to 0.7 mmol of DTPA) of the title compound from Example 18b) are dissolved in analogy as described for Example 14c) with 0.18 g (0.72 mmol) of an aqueous gadolinium chloride solution 10 ml distilled water, complexed. After working up and freeze-drying, 1.05 g (97.3% of theory) of the title compound is obtained as an amorphous powder.
Water content: 5.67%

Elemental analysis (calculated on anhydrous substance):
calc .: C 43.59; H 5.77; N 12.53; S 2.98; Gd 10.42; Na 1.52;
Found: C 43.61; H 5.81; N 12.49; S 2.94; Gd 10.48; Na 1.47;

The molar element ratio of gadolinium to sulfur results in thus an average degree of loading of the polymer of 41% Gd-DPTA and 59% D-galactose (corresponding to 10 Gd-DTPA units and 14-D-galactose residues per molecule). There are no ninhydrin reactions in the title compound free amino functions more detectable.

Example 19 24-Starburst dendrimer (Gd-DTPA amide) ₁₀ - [1- (4-thioureioureidophenyl) -α- D-mannopyranosyl] ₁₄ conjugate a) 24-Starburst dendrimer (DTPA amide) ₁₀ - [1- (4-thioureidophenyl) -α- D-mannopyranosyl] ₁₄ conjugate

In an analogous manner to that described for Example 14b), the reaction of 0.95 g (0.096 mmol; corresponding to 1.344 mmol of free amine functions) of the title compound from Example 18a) with 1.29 g (94.6 mmol) of p-isothiocyanato phenyl -α-D-mannopyranose after working up 1.29 g (94.6% of theory) of an amorphous and colorless solid.
Water content: 6.88%

Elemental analysis (calculated on anhydrous substance):
calc .: C 46.25; H 6.13 N 13.31; S 3.16; Na 6.41;
Found: C 46.09; H 6.07; N 13.33; S 3.24; Na 6.51;

The ninhydrin reaction of the title compound is negative, i. H. the polymer Product does not contain any free amino functions. From the percentage sulfur The elemental analysis of the title compound gives a degree of loading of the polymer with 59% D-mannose (corresponding to 14 mannose residues per mo lekül).

b) 24-Starburst dendrimer (Gd-DTPA amide) ₁₀ - [1- (4-thioureioureidophenyl) -α- D-mannopyranosyl] ₁₄ conjugate

1.1 g (0.077 mmol; corresponding to 0.77 mmol of DTPA) of the title compound from Example 19a) are analogous to that described for Example 14c) with 194.2 mg (0.79 mmol) of gadolinium chloride, dissolved in 10 ml of distilled water , complexed. After working up and freeze-drying, 1.14 g (98.6% of theory) of the title compound is obtained as an amorphous powder.
Water content: 4.72%

Elemental analysis (calculated on anhydrous substance):
calc .: C 43.55; H 5.77; N 12.53; S 2.98; Gd 10.42; Na 1.52;
Found: C 43.61; H 5.82; N 12.48; S 3.04; Gd 10.38; Na 1.49;

The molar element ratio of gadolinium to sulfur results in thus an average degree of loading of the polymer of 41% Gd-DPTA and 59 D-mannose (corresponding to 10 Gd-DTPA units and 14-D-mannose residues per molecule). There are no ninhydrin reactions in the title compound free amino functions more detectable.  

Example 20 24-Starburst dendrimer (Gd-DTPA amide) ₁₀ - [1- (4-thioureidophenyl) -β-D-glucopyranosyl] ₁₄ conjugate a) 24-Starburst dendrimer (DTPA amide) ₁₀ - (1- (4-thioureidophenyl) -β-D-glucopyranosyl] ₁₄ conjugate

In an analogous manner to that described for Example 14b), the reaction of 1.2 g (0.12 mmol; corresponding to 1.21 mmol of free amine functions) of the title compound from Example 18a) gives 1.92 g (6.03 mmol) ) p-isothiocyanatophenyl-β-D-glucopyranose after working up 1.59 g (93.6% of theory) of an amorphous and colorless solid.
Water content: 5.97%

Elemental analysis (calculated on anhydrous substance):
calc .: C 46.25; H 6.13; N 13.31; S 3.16; Na 6.27;
Found: C 46.20; H 6.21; N 13.32; S 3.08; Na 6.37;

The ninhydrin reaction of the title compound is negative, i. H. the polymer Product does not contain any free amino functions. From the percentage sulfur The elemental analysis of the title compound gives a degree of loading of the polymer with 59% D-glucose (corresponding to 14 glucose residues per mole cool).

b) 24-Starburst dendrimer (Gd-DTPA amide) ₁₀ - [1- (4-thioureidophenyl) -β-D-glucopyranosyl] ₁₄ conjugate

1.40 g (0.089 mmol; corresponding to 0.89 mmol of DTPA) of the title compound from Example 20a) are in analogy as described for Example 14c) with 219 mg (0.90 mmol) of gadolinium chloride, dissolved in 10 ml of distilled water plexed. After working up and freeze-drying, 1.32 g (98.7% of theory) of the title compound is obtained as an amorphous powder.
Water content: 7.33%

Elemental analysis (calculated on anhydrous substance):
calc .: C 43.55; H 5.77; N 12.53; S 2.98; Gd 10.42; Na 1.52;
Found: C 43.62; H 5.80; N 12.49; S 2.96; Gd 10.47; Na 1.58;

The molar element ratio of gadolinium to sulfur results in thus an average degree of loading of the polymer of 41% Gd-DPTA and 59% D-glucose (corresponding to 10 Gd-DTPA units and 14-D-glucose residues per Molecule). There are no free ninhydrin reactions in the title compound Amino functions more detectable.

Example 21 24-Starburst dendrimer (Gd-DTPA amide) ₁₀ - [1- (4-thioureidophenyl) -α-L-fucopyranosyl] ₁₄ conjugate a) 24-Starburst dendrimer (DTPA amide) ₁₀ - [1- (4-thioureidophenyl) -α-L-fucopyranosyl] ₁₄ conjugate

In an analogous manner as described for Example 14b), the reaction of 1.0 g (0.10 mmol; corresponding to 1.4 mmol of free amine functions) of the title compound from Example 18a) with 1.50 g (5.03 mmol) ) p-isothiocyanatophenyl-α -L-fucopyranose after working up 1.3 g (94.2% of theory) of an amorphous and colorless solid.
Water content: 6.08%

Elemental analysis (calculated on anhydrous substance):
calc .: C 47.00; H 6.23; N 13.53; S 3.21; Na 6.58;
Found: C 46.94; H 6.19; N 13.55; S 3.25; Na 6.60;

The ninhydrin reaction of the title compound is negative, i. H. the polymer Product does not contain any free amino functions. From the percentage sulfur The elemental analysis of the title compound gives a degree of loading of the polymer with 59% L-fucose (corresponding to 14 fucose residues per molecule).  

b) 24-Starburst dendrimer (Gd-DTPA amide) ₁₀ - [1- (4-thioureidophenyl) -α- L-fucopyranosyl] ₁₄ conjugate

1.1 g (0.078 mmol; corresponding to 0.78 mmol of DTPA) of the title compound from Example 21a) are combined in a manner analogous to that described for Example 14c) with 197 mg (0.79 mmol) of gadolinium chloride, dissolved in 10 ml of distilled water xiert. After working up and freeze-drying, 1.14 g (98.6% of theory) of the title compound is obtained as an amorphous powder.
Water content: 6.97%

Elemental analysis (calculated on anhydrous substance):
calc .: C 44.20; H 5.86; N 12.72; S 3.02; Gd 10.58; Na 1.55;
Found: C 44.28; H 5.88; N 12.69; S 2.99; Gd 10.62; Na 1.51;

The molar element ratio of gadolinium to sulfur results in thus an average degree of loading of the polymer of 41% Gd-DPTA and 59% D-glucose (corresponding to 10 Gd-DTPA units and 14-D-glucose residues per Molecule). There are no free ones in the title compound by means of a ninhydrin reaction Amino functions more detectable.

Example 22 24-Starburst dendrimer (Gd-DTPA amide) ₁₀ - [1- (4-thioureidophenyl) -N-acetyl-β-D-glucosaminopyranosyl] ₁₄ conjugate a) 24-Starburst dendrimer (DTPA amide) ₁₀ - [1- (4-thioureidophenyl) -N-acetyl-β-D-glucosaminopyranosyl] ₁₄ conjugate

In an analogous manner as described for Example 14b), the reaction of 1.0 g (0.10 mmol; corresponding to 1.4 mmol of free amine functions) of the title compound from Example 18a) with 2.50 g (7.04 mmol) ) p-isothiocyanatophenyl-β-D-glucopyranose after working up 1.4 g (95% of theory) of an amorphous and colorless solid.
Water content: 8.06%

Elemental analysis (calculated on anhydrous substance):
calc .: C 46.73; H 6.18; N 14.12; S 3.04; Na 6.22;
Found: C 46.69; H 6.21; N 14.20; S 3.00; Na 6.24;

The ninhydrin reaction of the title compound is negative, i. H. the polymer Product does not contain any free amino functions. From the percentage sulfur The elemental analysis of the title compound gives a degree of loading of the polymer with 59% N-acetyl-D-glucosamine (corresponding to 14 N-acetyl-D- Residues of glucosamine per molecule).

b) 24-Starburst dendrimer (Gd-DTPA amide) ₁₀ - [1- (4-thioureidophenyl) -N-acetyl-β-D-glucosaminopyranosyl] ₁₄ conjugate

1.6 g (0.10 mmol; corresponding to 1.08 mmol of DTPA) of the title compound from Example 22a) are described in analogy as described for Example 14c) with 278 mg (1.10 mmol) of gadolinium chloride, dissolved in 10 ml of distilled water , complexed. After working up and freeze-drying, 1.54 g (98.7% of theory) of the title compound is obtained as an amorphous powder.
Water content: 5.51%

Elemental analysis (calculated on anhydrous substance):
calc .: C 44.10; H 5.83; N 13.33; S 2.87; Gd 10.04; Na 1.47;
Found: C 44.19; H 5.91; N 13.41; S 2.80; Gd 10.08; Na 1.44;

The molar element ratio of gadolinium to sulfur results in thus an average degree of loading of the polymer of 41% Gd-DPTA and 59% N-acetyl-D-glucosamine (corresponding to 10 Gd-DTPA units and 14 N-acetyl D-glucosamine residues per molecule). In the title compound are using ninhydrin reaction no free amino functions detectable.

Example 23 36-cascade amine (Gd-Gly-Me-DOTA amide) ₁₆ - (α-D-mannopyranosyl-1-thio-3-propionic acid amide) ₂₀ conjugate a) 36-cascade amine (Gly-Me-DOTA-tri-t.-butyl ester amide) ₁₁

20.0 g (26.8 mmol) Gly-Me-DOTA-tri-tert.butyl ester (title compound from Bei game 31f) are at room temperature in 100 ml of absolute dimethylformamide dissolved and mixed with 3.7 g (32.16 mmol) of N-hydroxysuccinimide. The clear reaction mixture is then cooled to 0 ° C. and at this temperature in portions with a total of 6.6 g (32.16 mmol) of dicyclo hexylcarbodiimide added. After a reaction time of 30 minutes at 0 ° C and The precipitated dicyclohexylurea is removed for 2.5 hours at room temperature filtered off. The filtrate thus obtained is now slowly added dropwise at room temperature to a stirred solution of 8.8 g (1.18 mmol; corresponding to 42.8 mmol free amino functions) 36 cascade polyamine and 4.3 g (44.8 mmol) Triethylamine in 60 ml of absolute DMF. After a response time of 12 hours at room temperature is evaporated to dryness in vacuo and the ver remaining residue with 250 ml of dichloromethane. After filtering off of insoluble parts, the organized phase becomes successively with 5% Hydrochloric acid, saturated sodium bicarbonate solution twice with water washed. After drying the organic product phase over sodium sulfate is filtered and the solvent removed in vacuo. You get 12.5 g (86.2% of theory; based on the polyamine used) of a slightly yellow colored oil, which without further cleaning immediately for the subsequent Synthetic steps are used. After quantitative ninhydrin reaction the above title compound has an average of 20 free amine functions per Molecule before.

Elemental analysis:
calc .: C 67.65; H 10.32; N 8.72;
Found: C 67.50; H 10.41; N 8.91;

b) 36-cascade amine (Gly-Me-DOTA-tri-t.-butyl ester amide) ₁₆ - [3- (2,3,4,6-tetra-O-acetyl-1-thio) thio) - α-D-mannopyranosyl-3-propione-amide] ₂₀ conjugate

Stirred solution of 3.5 g (0.28 mmol; corresponding to 5.6 mmol of free Amine functions) of the title compound from Example 23a) in 100 ml of absolute Dichloromethane is at room temperature with 708 mg (7.0 mmol) of triethylamine transferred. Then with a solution of 3.6 g (6.8 mmol) of 3- (2,3,4,6- Tetra-O-acetyl-1-thio-α-D-mannopyranosyl) propionic acid N-hydroxysuccinimide  ester (representation in analogy to: J. Haensler et al., Bioconjugate Chem 4, 85, (1993); Chipowsky, S., and Lee, Y. C (1973) "Synthesis of 1-Thio-aldosides"; Carbohydrate Research 31, 339-346) in 60 ml dichloromethane dropwise transferred. After a reaction time of 12 hours at room temperature made up to a total volume of 500 ml with dichloromethane. The organi cal phase is successively twice with water and twice with saturated Washed sodium bicarbonate solution and dried over sodium sulfate. After filtering, the solvent is removed to dryness in vacuo drawn. 5.25 g (90.4% of theory) of the title compound are obtained as a colorless oil. The ninhydrin reaction of the title compound is negative, i. H. are in the polymer free amino functions no longer detectable.

Elemental analysis:
calc .: C 59.99; H 8.38; N 5.19; S 3.09;
Found: C 60.41; H 8.52; N 5.08; S 3.00;

From the percentage sulfur value of the elemental analysis of the title compound there is an average degree of loading of the polymer with 55.5% D-mannose tetraacetate (equivalent to 20 D-mannose residues per molecule).

c) 36-cascade amine (Gly-Me-DOTA amide) ₁₆ - (α-D-mannopyranosyl-1-thio-3-propionic acid amide) ₂₀ conjugate

5.0 g (0.24 mmol) of the title compound from Example 23b) 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 concentrated 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 at room temperature with 50 ml of a 32% aqueous ammonia solution. 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 brought to pH 3.0 by adding 10% hydrochloric acid and 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: 2.69 g (94.1% of theory) as an amorphous and colorless powder
Water content: 7.78%

Elemental analysis (calculated on anhydrous substance):
calc .: C 47.03; H 6.62; N 9.06; S 5.19;
Found: C 46.98; H 6.73; N 9.10; S 5.24;

d) 36-cascade amine (Gd-Gly-Me-DOTA amide) ₁₆ - (α-D-mannopyranosyl-1-thio-3-propionic acid amide) ₂₀ conjugate

2.0 g (0.16 mmol; corresponding to 2.6 mmol of Gly-Me-DOTA) of the title compound solution from Example 23c) are dissolved in 70 ml of distilled water and the pH The value of the solution is determined by dropwise addition with 10% aqueous salt acidity set to 5.0. Finally, 709 mg are added at room temperature (2.7 mmol) gadolinium chloride, dissolved in 10 ml of distilled water, and the reaction solution is stirred at 60 ° C. for 12 hours. At room temperature the reaction solution by adding 1 molar sodium hydroxide solution to pH 7.2 ge brought and made up with distilled water to a total volume of 500 ml fills. After three times ultrafiltration against distilled water over a YM3 Ultrafiltration membrane (AMICON®) freezes the remaining residue dried. 2.31 g (97.5% of theory) of the title compound are obtained as an amorphous, colorless powder with a water content of 6.7%.

Elemental analysis (calculated on anhydrous substance):
calc .: C 39.24; H 5.11; N 7.56; S 4.33; Gd 16.98;
Found: C 39.42; H 5.21; N 7.52; S 4.39; Gd 17.02;

The molar element ratio of sulfur to gadolinium results thus an average degree of loading of the polymer of 44% Gd-Gly-Me-DOTA and 56% D-mannose (corresponding to 16 Gd-Gly-Me-DOTA units and 20-D- Mannose residues per molecule). In the title link are using ninhydrin crack tion no free amino functions detectable.  

Example 24 36-cascade amine (Gd-Gly-Me-DOTA amide) ₁₆ - (β-D-glucopyranosyl-1-thio-3-propionic acid amide) ₂₀ conjugate a) 36-cascade amine (Gd-Gly-Me-DOTA-tri-tert-butyl ester amide) ₁₆ - [- (2,3,4,6-tetra-O-acetyl-1-thio) -β-D -glucopyranosyl-3-propionic acid amide)] ₂₀ conjugate

In an analogous manner as described for Example 23b), the conversion of 3.5 g (0.28 mmol; corresponding to 5.6 mmol of free amine functions) of the title Compound from Example 23a) with 3.6 g (6.8 mmol) of 3- (2,3,4,6-tetra-O-acetyl-1- thio-β-D-glucopyranosyl) propionic acid N-hydroxysuccinimide ester (shown in Analogy to: J. Haensler et al., Bioconjugate Chem. 4, 85, (1993); Chipowsky, S., and Lee, Y.C. (1973), Synthesis of 1-Thio-aldosides; Carbohydrate Re search 31, 339-346) 5.4 g (93% of theory) of the title compound as a colorless oil.

Elemental analysis:
calc .: C 59.99; H 8.38; N 5.19; S 3.09;
Found: C 60.32; H 8.30; N 5.09; S 3.18;

The title compound shows a negative quantitative ninhydrin reaction. From the percentage sulfur value of the elementary analysis results in an average Bela degree of polymer with 20 D-glucose residues per molecule.

b) 36-cascade amine (Gly-Me-DOTA amide) ₁₆ - (β-D-glucopyranosyl-1-thio-3-propionic acid amide) ₂₀ conjugate

In an analogous manner as described for Example 23c), the deprotection of 5 g (0.24 mmol) of the title compound from Example 24b) after freeze-drying leads to the formation of 2.6 g (88.6% of theory) of the above title compound as an amorphous powder.
Water content: 6.03%

Elemental analysis (calculated on anhydrous substance):
calc .: C 47.03; H 6.62; N 9.06; S 5.19;
Found: C 47.12; H 6.71; N 9.02; S 5.30;

c) 36-cascade amine (Gd-Gly-Me-DOTA amide) ₁₆ - (β-D-glucopyranosyl-1-thio-3-propionic acid amide) ₂₀ conjugate

In an analogous manner as described for Example 23d), the complexation of 2.0 g (0.16 mmol; corresponding to 2.6 mmol of Gly-Me-DOTA) of the title compound from Example 24b) with 709 mg (2.7 mmol) gadolinium chloride, dissolved in 10 ml of distilled water, to form the title compound mentioned above.
Yield: 2.35 g (98% of theory) as a colorless and amorphous powder
Water content: 5.2%

Elemental analysis (calculated on anhydrous substance):
calc .: C 39.24; H 5.11; N 7.56; S 4.33; Gd 16.98;
Found: C 39.61; H 5.20; N 7.49; S 4.41; Gd 16.94;

Average degree of loading per molecule of the above title compound:
16 Gd-Gly-Me-DOTA units (degree of polymer loading of 44.4%)
and 20 D-glucopyranosyl residues (degree of polymer loading of 55.6%)

The polymer loading levels were calculated from the molar element ratios determined from sulfur to gadolinium.

Example 25 36-cascade amine (Gd-Gly-Me-DOTA amide) ₁₆ - (β-D-galactopyranosyl-1-thio-3-propionic acid amide) ₂₀ conjugate a) 36-cascade amine (Gd-Gly-Me-DOTA-tri-tert-butyl ester amide) ₁₆ - [3- (2,3,4,6-tetra-O-acetyl-1-thio) -β- D-galactopyranosyl-3-propionic acid amide)] ₂₀ conjugate

In an analogous manner as described for Example 23b), the conversion of 3.5 g (0.28 mmol; corresponding to 5.6 mmol of free amine functions) of the title  Compound from Example 23a) with 3.6 g (6.8 mmol) of 3- (2,3,4,6-tetra-O-acetyl-1- thio-β-D-galactopyranosyl) propionic acid N-hydroxysuccinimide ester [representation in analogy to: J. Haensler et al., Bioconjugate Chem. 4, 85, (1993); Chipowsky, S., and Lee, Y. C (1973), "Synthesis of 1-Thio-aldosides; Carbohydrate Re search 31, 339-346] 5.1 g (87.8% of theory) of the title compound as a colorless oil.

Elemental analysis:
calc .: C 59.99; H 8.38; N 5.19; S 3.09;
Found: C 60.03; H 8.46; N 5.24; S 3.14;

The title compound shows a negative ninhydrin reaction. From the percent len sulfur value of the elementary analysis results in an average degree of loading of the polymer with 20 D-galactose residues per molecule.

b) 36-cascade amine (Gly-Me-DOTA amide) ₁₆ - (β-D-galactopyranosyl-1-thio-3-propionic acid amide) ₂₀ conjugate

In an analogous manner as described for Example 23c), the deprotection of 5.0 g (0.24 mmol) of the title compound from Example 25b) after freeze drying leads to the formation of 2.64 g (90.2% of theory). ) the above-mentioned title compound as an amorphous powder.
Water content: 8.3%

Elemental analysis (calculated on anhydrous substance):
calc .: C 47.03; H 6.62; N 9.06; S 5.19;
Found: C 47.12; H 6.71; N 9.00; S 5.07;

c) 36-cascade amine (Gd-Gly-Me-DOTA amide) ₁₆ - (β-D-galactopyranosyl-1-thio-3-propionic acid amide) ₂₀ conjugate

In an analogous manner as described for Example 23d), the complexation of 2.5 g (0.2 mmol; corresponding to 3.2 mmol of Gly-Me-DOTA) of the title compound from Example 24b) with 867 mg (3.3 mmol) gadolinium chloride, dissolved in 10 ml of distilled water, to form the title compound mentioned above.
Yield: 2.88 g (97.3% of theory) as a colorless and amorphous powder
Water content: 9.3%

Elemental analysis (calculated on anhydrous substance):
calc .: C 39.24; H 5.11; N 7.56; S 4.33; Gd 16.98;
Found: C 39.21; H 5.04; N 7.63; S 4.41; Gd 17.07;

Average degree of loading per molecule of the above title compound:
16 Gd-Gly-Me-DOTA units (degree of polymer loading of 43.8%)
and 20 D-galactopyranosyl residues (degree of polymer loading of 56.2%)

The polymer loading levels were calculated from the molar element ratios determined from sulfur to gadolinium.

Example 26 48-cascade amine (Gd-Gly-Me-DOTA amide) ₂₁ - (α-D-mannopyranosyl-1-thio-3-propionic acid amide) ₂₇ conjugate a) 48-cascade amine (Gly-Me-DOTA-tri-tert.butyl ester amide) ₂₁

20.0 g (26.8 mmol) Gly-Me-DOTA-tri-tert.butyl ester (title compound from Bei game 31f) are at room temperature in 100 ml of absolute dimethylformamide dissolved and mixed with 3.7 g (32.16 mmol) of N-hydroxysuccinimide. The clear reaction mixture is then cooled to 0 ° C. and at this temperature in portions with a total of 6.6 g (32.16 mmol) of dicyclo hexylcarbodiimide added. After a reaction time of 30 minutes at 0 ° C and The precipitated dicyclohexylurea is removed for 2.5 hours at room temperature filtered off. The filtrate thus obtained is now slowly added dropwise at room temperature to a stirred solution of 6.0 g (0.92 mmol; corresponding to 4.4 mmol free amino functions) 48-part cascade polyamine and 4.5 g (46.8 mmol) Triethylamine in 60 ml of absolute DMF. After a response time of 12 hours at room temperature is evaporated to dryness in vacuo and the ver remaining residue with 250 ml of dichloromethane. After filtering off of insoluble parts, the organized phase becomes successively with 5% Hydrochloric acid, saturated sodium bicarbonate solution twice with water washed. After drying the organic product phase over sodium sulfate is filtered and the solvent removed in vacuo. You get  13.75 g (83.2% of theory; based on the polyamine used) of a slightly yellow colored oil, which without further cleaning immediately for the subsequent Synthetic steps are used. After quantitative ninhydrin reaction the above title compound has an average of 27 free amine functions per Molecule before.

Elemental analysis:
calc .: C 63.35; H 9.61; N 16.36;
Found: C 63.18; H 9.70; N 16.44;

b) 48-cascade amine (Gd-Gly-Me-DOTA-tri-tert-butyl ester amide) ₂₁ - [3- (2,3,4,6-tetra-O-acetyl-1-thio) -α- D-mannopyranosyl-3-propion-amide] ₂₇ - conjugate

A stirred solution of 3.5 g (0.19 mmol; corresponding to 5.13 mmol of free Amine functions) of the title compound from Example 26a) in 100 ml of absolute Dichloromethane is at room temperature with 657 mg (6.5 mmol) of triethylamine transferred. Then with a solution of 3.3 g (6.3 mmol) of 3- (2,3,4,6- Tetra-O-acetyl-1-thio-α-D-mannopyranosyl) propionic acid N-hydroxysuccinimide ester added dropwise in 60 ml of dichloromethane. After a response time of 12 hours at room temperature is made up to a total with dichloromethane volume of 500 ml filled. The organic phase becomes two in succession times with water and twice with saturated sodium bicarbonate solution washed and dried over sodium sulfate. After filtering it will Stripped solvent to dryness in vacuo. 5.07 g (92.6 % d. Th.) Of the title compound as a colorless oil. The quantitative ninhydrin freak tion of the title link is negative, i. H. there are no free ones in the polymer Amino functions more detectable.

Elemental analysis:
calc .: C 58.65; H 7.98; N 10.21; S 3.00;
Found: C 58.90; H 8.08; N 10.14; S 2.96;

From the percentage sulfur value of the elemental analysis of the title compound there is an average degree of loading of the polymer with 56.2% D-mannose tetraacetate (equivalent to 27 D-mannose residues per molecule).  

c) 48-cascade amine (Gly-Me-DOTA amide) ₂₁ - (α-D-mannopyranosyl-1-thio-3-propionic acid amide) ₂₇ conjugate

5.0 g (0.17 mmol) of the title compound from Example 26b) 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 concentrated 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 at room temperature with 50 ml of a 32% aqueous ammonia solution. 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 brought to pH 3.0 by adding 10% hydrochloric acid and 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: 3.47 g (90.6% of theory) as an amorphous and colorless powder
Water content: 9.11%

Elemental analysis (calculated on anhydrous substance):
calc .: C 51.08; H 7.12; N 13.32; S 3.92;
Found: C 50.97; H 7.21; N 13.30; S 3.86;

d) 48-cascade amine (Gd-Gly-Me-DOTA amide) ₂₁ - (α-D-mannopyranosyl-1-thio-3-propionic acid amide) ₂₇ conjugate

3.0 g (0.136 mmol; corresponding to 285 mmol of Gly-Me-DOTA) of the title compound solution from Example 26c) are dissolved in 70 ml of distilled water and the pH The value of the solution is determined by dropwise addition with 10% aqueous salt acidity set to 5.0. Finally, 787 mg are added at room temperature (3.0 mmol) gadolinium chloride, dissolved in 10 ml of distilled water, and the reaction solution is stirred at 60 ° C. for 12 hours. At room temperature  the reaction solution by adding 1 molar sodium hydroxide solution to pH 7.2 ge brought and made up with distilled water to a total volume of 500 ml fills. After three times ultrafiltration against distilled water over a YM3 Ultrafiltration membrane (AMICON®) freezes the remaining residue dried. 3.31 g (96.3% of theory) of the title compound are obtained as an amorphous, colorless powder with a water content of 4.9%.

Elemental analysis (calculated on anhydrous substance):
calc .: C 44.54; H 5.96; N 11.62; S 3.42; Gd 13.04;
Found: C 44.59; H 6.04; N 11.55; S 3.41; Gd 13.10;

The molar element ratio of sulfur to gadolinium results thus an average degree of loading of the polymer of 43.8% Gd-Gly-Me-DOTA and 56.2% D-mannose (corresponding to 21 Gd-Gly-Me-DOTA units and 27- D-mannose residues per molecule). In the title compound are using ninhydrin reaction no free amino functions detectable.

Example 27 48-cascade amine (Gd-Gly-Me-DOTA amide) ₂₁ - (β-D-glucopyranosyl-1-thio-3-propionic acid amide) ₂₇ conjugate a) 48-cascade amine (Gd-Gly-Me-DOTA-tri-tert-butyl ester amide) ₂₁ - [3- (2,3,4,6-tetra-O-acetyl-1-thio) -β- D-glucopyranosyl-3-propionamide] ₂₇ conjugate

In an analogous manner as described for Example 26b), the implementation of 3.5 g (0.19 mmol; corresponding to 5.13 mmol of free amine functions) the title Compound from Example 26a) with 3.3 g (6.3 mmol) of 3- (2,3,4,6-tetra-O-acetyl-1- thio-β-D-glucopyranosyl) propionic acid N-hydroxysuccinimide ester 5.57 g (93 d. Th.) Of the title compound as a colorless oil.

Elemental analysis:
calc .: C 58.65; H 7.98; N 10.21; S 3.00;
Found: C 58.68; H 8.10; N 10.20; S 3.07;

The title compound shows a negative ninhydrin reaction. From the percent len sulfur value of the elementary analysis results in an average degree of pollution of the polymer with 27 D-glucose residues per molecule.

b) 48-cascade amine (Gly-Me-DOTA amide) ₂₁ - (β-D-glucopyranosyl-1-thio-3-propionic acid amide) ₂₇ conjugate

In an analogous manner as described for Example 26c), the deprotection of 5 g (0.17 mmol) of the title compound from Example 27a) after freeze-drying leads to the formation of 2.23 g (84.7% of theory) of the above title compound as an amorphous powder.
Water content: 7.44%

Elemental analysis (calculated on anhydrous substance):
calc .: C 51.08; H 7.12; N 13.32; S 3.92;
Found: C 51.11; H 7.10; N 13.26; S 4.01;

c) 48-cascade amine (Gd-Gly-Me-DOTA amide) ₂₁ - (β-D-glucopyranosyl-1-thio-3-propionic acid amide) ₂₇ conjugate

In an analogous manner to that described for Example 26d), the complexation of 3.0 g (0.136 mmol; corresponding to 2.85 mmol of Gly-Me-DOTA) of the title compound from Example 27b) leads to 787 mg (3.0 mmol) Gadolinium chloride dissolved in 10 ml of distilled water to form the title compound mentioned above.
Yield: 3.36 g (98% of theory) as a colorless and amorphous powder
Water content: 4.41%

Elemental analysis (calculated on anhydrous substance):
calc .: C 44.54; H 5.96; N 11.62; S 3.42; Gd 13.04;
Found: C 44.39; H 5.88; N 11.74; S 3.40; Gd 13.00;

Average degree of loading per molecule of the above title compound:
21 Gd-Gly-Me-DOTA units (degree of polymer loading of 43.8%)
and 27 D-glucopyranosyl residues (degree of polymer loading of 56.2%)

The polymer loading levels were calculated from the molar element ratios determined from sulfur to gadolinium.  

Example 28 48-cascade amine (Gd-Gly-Me-DOTA amide) ₂₁ - (β-D-galactopyranosyl-1-thio-3-propionic acid amide) ₂₇ conjugate a) 48-cascade amine (Gly-Me-DOTA-tri-tert-butyl ester amide) ₂₁ - [3- (2,3,4,6-tetra-O-acetyl-1-thio) -β-D- galactopyranosyl-3-propione-amide] ₂₇ conjugate

In an analogous manner as described for Example 26b), the implementation of 3.5 g (0.19 mmol; corresponding to 5.13 mmol of free amine functions) the title Compound from Example 26a) with 3.3 g (6.3 mmol) of 3- (2,3,4,6-tetra-O-acetyl-1- thio-β-D-galactopyranosyl) propionic acid N-hydroxysuccinimide ester 5.23 g (90.2 % d. Th.) Of the title compound as a colorless oil.

Elemental analysis:
calc .: C 58.65; H 7.98; N 10.21; S 3.00;
Found: C 58.66; H 8.00; N 10.31; S 3.11;

The title compound shows a negative ninhydrin reaction. From the percent len sulfur value of the elementary analysis results in an average degree of loading of the polymer with 27 D-galactose residues per molecule.

b) 48-cascade amine (Gly-Me-DOTA amide) ₂₁ - (β-D-galactopyranosyl-1-thio-3-propionic acid amide) ₂₇ conjugate

In an analogous manner to that described for Example 26c), the deprotection of 5.0 g (0.17 mmol) of the title compound from Example 28a) leads to the formation of 3.48 g (91.4% of theory) after freeze-drying. ) the above-mentioned title compound as an amorphous powder.
Water content: 6.79%

Elemental analysis (calculated on anhydrous substance):
calc .: C 51.08; H 7.12; N 13.32; S 3.92;
Found: C 51.12; H 7.19; N 13.27; S 3.99;

c) 48-cascade amine (Gd-Gly-Me-DOTA amide) ₂₁ - (β-D-galactopyranosyl-1-thio-3-propionic acid amide) ₂₇ conjugate

In an analogous manner as described for Example 26d), the complexation of 3.0 g (0.136 mmol; corresponding to 2.85 mmol of Gly-Me-DOTA) of the title compound from Example 28b) with 787 mg (3.3 mmol) Gadolinium chloride dissolved in 10 ml of distilled water to form the title compound mentioned above.
Yield: 3.32 g (96.9% of theory) as a colorless and amorphous powder
Water content: 8.70%

Elemental analysis (calculated on anhydrous substance):
calc .: C 44.54; H 5.96; N 11.62; S 3.42; Gd 13.04;
Found: C 44.50; H 5.94; N 11.58; S 3.37; Gd 12.98;

Average degree of loading per molecule of the above title compound:
21 Gd-Gly-Me-DOTA units (degree of polymer loading of 43.6%)
and 27 D-galactopyranosyl residues (degree of polymer loading of 56.4%)

The polymer loading levels were calculated from the molar element ratios determined from sulfur to gadolinium.

Example 29 48-Starburst dendrimer (Gd-DTPA amide) ₁₉ - (O-β-D-galactopyranosyl (1 → 4) - D-glucono-1,5-lactone amide) ₂₉ conjugate a) 48-Starburst dendrimer (DTPA amide) ₁₉ - (O-β-D-galactopyranosyl (1 → 4) -D-glucono-1,5-lactone amide) ₂₉ conjugate

To a stirred solution of 4.0 g (0.20 mmol; corresponding to 6.0 mmol of free amino functions) of the title compound from Example 14a) are suspended in 40 ml of absolute dimethyl sulfoxide at room temperature. Then a solution of 13.3 g (37.2 mmol, corresponding to 6.26 g lactone per amino function present) O-β-D-galactopyranosyl- (1-4) -D-glucono-1,5-lactone ( Representation according to (a) Williams, TJ; Plessas, NR, Goldstein, IJ Carbohydr. Res. 1978, 67, CI. (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 40 ml of absolute dimethyl sulfoxide at 22 ° C. added dropwise. The reaction solution thus obtained is then stirred at 40 ° C. for 14 hours. For working up, 500 ml of absolute methanol are added at room temperature, and the resulting colorless precipitate is filtered off with a G4 frit and washed thoroughly with a total of 250 ml of absolute methanol. The solid obtained in this way is then dissolved in 500 ml of distilled water and ultrafiltered a total of three times through an ultrafiltration membrane (YM-3, Amicon, 3 kD exclusion limit). The remaining residue is completely dissolved in distilled water and freeze-dried.
Yield: 5.42 g (92.7% of theory) as a colorless lyophilisate
Water content: 8.34%

Elemental analysis (calculated on anhydrous substance):
calc .: C 44.23; H 6.31; N 11.66; Na 5.96;
Found: C 44.41; H 6.28; N 11.60; Na 6.03;

After elemental analysis as well as colorimetric titration with 0.0025 molar Gadolinium sulfate standard solution in the presence of xylenol orange as an indicator, he there is a DTPA loading level of the polymer of 39.4% (corresponding to 19th DTPA units per molecule). After ninhydrin reaction are mentioned in the above No more amino functions can be detected in the title compound.

b) 48-Starburst dendrimer (Gd-DTPA amide) ₁₉ - (O-β-D-galactopyranosyl (1 → 4) -D-glucono-1,5-lactone-amide) ₂₉ conjugate

4.0 g (0.136 mmol; corresponding to 2.6 mmol of DTPA) of the title compound from Example 29a) are combined in a manner analogous to that described for Example 1c) with 691 mg (2.65 mmol) of gadolinium chloride, dissolved in 10 ml of distilled water xiert. After working up and freeze-drying, 4.14 g (98.7% of theory) of the title compound are obtained as an amorphous powder.
Water content: 6.94%

Elemental analysis (calculated on anhydrous substance):
calc .: C 41.84; H 5.96; N 11.03; Gd 9.64; Na 1.41;
Found: C 41.90; H 6.03; N 10.97; Gd 9.61; Na 1.39;

The ninhydrin reaction as well as the TNBS method of the title compound are reasonable fen negative, d. H. the polymeric product contains no free amino functions.

Example 30 24-Starburst dendrimer (Gd-DTPA amide) ₁₀ - (O-β-D-galactopyranosyl- (1 → 4) -D-glucono-1,5-lactone amide) ₁₄ conjugate a) 24-Starburst dendrimer (DTPA amide) ₁₀ - (O-β-D-galactopyranosyl (1 → 4) -D-glucono-1,5-lactone amide) ₁₄ conjugate

To a stirred solution of 4.0 g (0.4 mmol), corresponding to 5.7 mmol of free amino functions) of the title compound from Example 18a), are suspended in 40 ml of absolute dimethyl sulfoxide at room temperature. Then a solution of 12.66 g (35.4 mmol, corresponding to 6.2 g lactone per amino function present) O-β-D-galactopyranosyl- (1-4) -D-glucono-1,5-lactone in 40 ml of absolute dimethyl sulfoxide added dropwise at 22 ° C. The reaction solution thus obtained is then stirred at 40 ° C. for 14 hours. For working up, 500 ml of absolute methanol are added at room temperature and the resulting colorless precipitate is filtered off with a G4 frit and washed well with a total of 200 ml of absolute methanol. The solid obtained in this way is then dissolved in 350 ml of distilled water and ultrafiltered a total of three times through an ultrafiltration membrane (YM-3, Amicon, 3 kD exclusion limit). The remaining residue is completely dissolved in distilled water and freeze-dried.
Yield: 5.6 g (96.0% of theory) as a colorless lyophilisate
Water content: 10.2%

Elemental analysis (calculated on anhydrous substance):
calc .: C 43.95; H 6.37; N 11.63; Na 6.31;
Found: C 44.06; H 6.22; N 11.59; Na 6.40;

After elemental analysis as well as colorimetric titration with 0.0025 molar Gadolinium sulfate standard solution in the presence of xylenol orange as an indicator, the polymer has a DTPA loading level of 41.5% (correspondingly 10 DTPA units per molecule).

After ninhydrin reaction as well as TNBS method are in the above Title compound no longer detectable amino functions.

b) 24-Starburst dendrimer (Gd-DTPA amide) ₁₀ - (O-β-D-galactopyranosyl- (1 → 4) -D-glucono-1,5-lactone amide) ₁₄ conjugate

5.0 g (0.343 mmol; corresponding to 10.34 mmol of DTPA) of the title compound from Example 30a) are described in analogy to that described for Example 1c) with 2.72 g (10.45 mmol) of gadolinium chloride, dissolved in 10 ml of distilled water , complexed. After working up and freeze-drying, 5.17 g (97.6% of theory) of the title compound are obtained as an amorphous powder.
Water content: 8.11%

Elemental analysis (calculated on anhydrous substance):
calc .: C 41.44; H 6.00; N 10.97; Gd 10.18; Na 1.49;
Found: C 41.51; H 5.96; N 11.03; Gd 10.24; Na 1.46;

The ninhydrin reaction as well as the TNBS method of the title compound are reasonable fen negative, d. H. the polymeric product contains no free amino functions.

Example 31 24-cascade amine (Gd-Gly-Me-DOTA amide) ₁₀ - [1- (4-thioureidophenyl) -β-D-galactopyranosyl] ₁₄ conjugate a) 24-cascade amine (Gd-Gly-Me-DOTA amide) ₁₀ conjugate

A stirred suspension of 5.0 g (7.95 mmol) of the title compound from example 31 h) 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-hydroxy succinimide and the reaction mixture is kept at this temperature for 1 hour. After cooling to 0 ° C, 4.52 g (23.85 mmol) of dicyclohexylcarbodiimide is added and the reaction solution is stirred for a further 1 hour at 0 ° C, followed by 12 hours at 22 ° C. The reaction solution thus obtained of the N-hydroxysuccinimide ester of the title compound from Example 31h) is now added dropwise at 22.degree. C. with a solution of 2.61 g (0.55 mmol, corresponding to 13.23 mmol of free amine functions, 24 cascade polyamine) 15 ml of absolute dimethyl sulfoxide are added and the mixture is stirred for a further 12 hours at room temperature, and the reaction solution is added dropwise in 1.5 l of acetone at 22 ° C., the title compound precipitating as a colorless precipitate, the precipitate is filtered off with suction, dissolved in 200 ml of distilled water and Ultrafilter three times through a YM3 ultrafiltration membrane (AMICON®), the remaining residue is dissolved in 300 ml of distilled water, filtered and freeze-dried.
Yield: 4.0 g (82.0% based on the polyamine used) as an amorphous and colorless powder.
Water content: 7.38%

Elemental analysis (calculated on anhydrous substance):
calc .: C 43.10; H 6.07; N 14.97; Gd 17.69;
Found: C 43.18; H 5.99; N 15.03; Gd 17.74;

According to elemental analysis, the degree of loading of the polymer is 41.5% Gd-Gly-Me-DOTA (corresponding to 10 Gd-Gly-Me-DOTA units per molecule). The determination of the free amino functions remaining in the polymer by means of Ninhydrin and the TNBS method provide an average of 58.1% of free aminogrup pen, d. H. There are 14 free amino functions in the polymer molecule.

b) 24-cascade amine (Gd-Gly-Me-DOTA amide) ₁₀ - [1- (4-thioureidophenyl) - β-D-galactopyranosyl] ₁₄ conjugate

In an analogous manner to that described for Example 1d), the reaction of 1.0 g (0.112 mmol; corresponding to 1.57 mmol of free amine functions) of the title compound from Example 31a) with 2.5 g (7.87 mmol; corresponding to 5 Mol equivalents based on the polyamine used p-isothiocyanatophenyl-β-D-galactopyranose after working up 1.33 g (93.2% of theory) of an amorphous and colorless solid.
Water content: 7.41%

Elemental analysis (calculated on anhydrous substance):
calc .: C 45.32; H 5.66; N 11.50; S 3.78; Gd 11.84;
Found: C 45.40; H 5.72; N 11.61; S 3.42; Gd 11.88;

The ninhydrin reaction as well as in the TNBS method of the title compound proceed negative, d. H. the polymeric product contains no free amino functions. Out the molar sulfur value of the elemental analysis of the title compound a degree of loading of the polymer with 58.3% D-galactopyranose (corresponding to 14 galactose residues per molecule) and 41.7% Gd-Gly-DOTA (corresponding to 10 Gd-Gly-Me-DOTA complexes per molecule).

c) N- (2-bromopropionyl) glycine benzyl ester

60.97 g (355.7 mmol) of α-bromopropionyl chloride are 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 in 500 ml of methylene chloride. The temperature is kept between 0 ° C-5 ° C. 1000 ml of 5% aqu. Hydrochloric acid and separates the organic phase. The organic phase is again with 500 ml of 5% aqu. Hydrochloric acid extracted, 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)

Elemental analysis:
calc .: C 48.02; H 4.70; N 4.67; Br 26.62;
Found: C 47.91; H 4.82; N 4.51; Br 26.47;

d) 1- [1-Benzyloxycarbonylmethylcarbamoyl) ethyl] -1,4,7,10-tetraazacyclodo decan

50 g (166.6 mmol) of the title compound from Example 31c) are added to 86.14 g (500 mmol) of 1,4,7,10-tetraazacyclododecane dissolved in 1000 ml of chloroform and the mixture is stirred for 24 hours at room temperature. It is extracted 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;

e) 10- [1-Benzyloxycarbonylmethylcarbamoyl) ethyl] -1,4,7,10-tetraazacyclodo tri-tert-butyl decane-1,4,7-triacetic acid, sodium bromide

To 50 g (127.7 mmol) of the title compound from Example 31d) and 54.14 g (510.8 mmol) of sodium carbonate in 500 ml of acetonitrile are added 82.20 g (421.4 mmol) of tert-butyl bromoacetate and stir for 12 hours at 60 ° C. 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;

f) 10- [1-carboxymethylcarbamoyl) ethyl] -1,4,7,10-tetraazacyclododecane 1,4,7-triacetic acid, tri-tert-butyl ester, sodium bromide

90 g (107.5 mmol) of the title compound from Example 31e) are dissolved in 1000 ml of isopropanol and 5 g of palladium catalyst (10% Pd / C) are added. It is hydrogenated overnight at room temperature. It is filtered off from the catalyst and the Fil stepped 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;

g) 10- [1-carboxymethylcarbamoyl) ethyl] -1,4,7,10-tetraazacyclododecane 1,4,7-triacetic acid

77 g (103.1 mmol) of the title compound from Example 31f) are dissolved in 500 ml of trifluoroacetic acid and stirred for 3 hours at room temperature. It is evaporated to dryness, the residue is taken up in 300 ml of water and the solution is placed on a column filled with Reillex® 425 PVP. You elute with water. The product-containing fractions are combined and evaporated to dryness, the residue is recrystallized 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):
calc .: C 47.99; H 6.99; N 14.73;
Found: C 47.83; H 7.12; N 14.55;

h) Gadolinium complex of 10- [1-carboxymethylcarbamoyl) ethyl] -1,4,7,10- tetraazacyclododecane-1,4,7-triacetic acid

15.27 g (42.06 mmol) of gadolinium oxide are added to 40 g (84.12 mmol) of the title compound from Example 31 g), dissolved in 400 ml of water, and the mixture is heated at 90 ° C. for 3 h. It is evaporated to dryness (vacuum) and the residue is crystallized from 90% aqu. Ethanol around. The crystals are filtered off with suction, washed once with ethanol, then with acetone and finally with dimethyl 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;

Example 32 24-cascade amine (Gd-Gly-Me-DOTA amide) ₁₀ - [1- (4-thioureidophenyl) - α-D-mannopyranosyl] ₁₄ conjugate a) 24-cascade amine (Gd-Gly-Me-DOTA amide) ₁₀ - [1- (4-thioureidophenyl) - α-D-mannopyranosyl] ₁₄ conjugate

In an analogous manner as described for Example 1b), the reaction of 1.0 g (0.11 mmol; corresponding to 1.57 mmol of free amine functions) of the title compound from Example 31g) with 2.5 g (7.87 mmol) ) p-isothiocyanatophenyl-α-D-mannopyranose after working up 1.38 g (94.7% of theory) of an amorphous and colorless solid.
Water content: 5.76%

Elemental analysis (calculated on anhydrous substance):
calc .: C 45.32; H 5.66; N 11.50; S 3.38; Gd 11.84;
Found: C 45.42; H 5.72; N 11.47; S 3.41; Gd 11.81;

The ninhydrin reaction as well as in the TNBS method of the title compound proceed negative, d. H. the polymeric product contains no free amino functions. Out the sulfur percentage of the elemental analysis of the title compound there is a degree of loading of the polymer with 56.8% D-mannopyranose (corresponding to 14 mannose residues per molecule) and 41.2% (corresponding to 10 Gd-Gly-Me-DOTA complexes per molecule).  

Example 33 24-cascade amine (Gd-Gly-Me-DOTA amide) ₁₀ - [1- (4-thioureidophenyl) -β-D-glucopyranosyl] ₁₄ conjugate a) 24-cascade amine (Gd-Gly-Me-DOTA amide) ₁₀ - [1- (4-thioureidophenyl) - β-dglucopyranosyl] ₁₄ conjugate

In an analogous manner to that described for Example 1b), the reaction of 1.2 g (0.13 mmol; corresponding to 1.89 mmol of free amine functions) of the title compound from Example 31g) gives 2.34 g (7.87 mmol) ) p-isothiocyanatophenyl-β-D-glucopyranose after working up 1.53 g (88.9% of theory) of an amorphous and colorless solid.
Water content: 6.44%

Elemental analysis (calculated on anhydrous substance):
calc .: C 46.05; H 5.85; N 11.68; S 7.44; Gd 12.03;
Found: C 45.96; H 5.90; N 11.71; S 3.40; Gd 12.10;

The ninhydrin reaction as well as in the TNBS method of the title compound proceed negative, d. H. the polymeric product contains no free amino functions. Out the sulfur percentage of the elemental analysis of the title compound there is a degree of loading of the polymer with 57.9% D-glucopyranose (corresponding to 14 glucose residues per molecule) and 42.1% (corresponding to 10 Gd-Gly-Me-DOTA complexes per molecule).

Example 34 24-cascade amine (Gd-Gly-Me-DOTA amide) ₁₀ - [1- (4-thioureidophenyl) -α-L-fucopyranosyl] ₁₄ conjugate a) 24-cascade amine (Gd-Gly-Me-DOTA amide) ₁₀ - [1- (4-thioureidophenyl) - α-L-fucopyranosyl] ₁₄ conjugate

In an analogous manner as described for Example 1b), the reaction of 1.0 g (0.112 mmol; corresponding to 1.57 mmol of free amine functions) of the titanium compound from Example 31a) with 2.34 g (7.87 mmol) p -Isothiocyanatophenyl- α-L-fucopyranose after working up 1.37 g (93.7% of theory) of an amorphous and colorless solid.
Water content: 4.92%

Elemental analysis (calculated on anhydrous substance):
calc .: C 46.05; H 5.85; N 11.68; S 7.44; Gd 12.03;
Found: C 45.96; H 5.90; N 11.71; S 3.40; Gd 12.10;

The ninhydrin reaction as well as in the TNBS method of the title compound proceed negative, d. H. the polymeric product contains no free amino functions. Out the sulfur percentage of the elemental analysis of the title compound there is a degree of loading of the polymer with 54.1% α-L-fucose (corresponding to 13 α-L-fucose residues per molecule) and 40.6% (corresponding 10 Gd-Gly-Me-DOTA complexes per molecule).

Example 35 a) 1- [4-Nitrophenyl] tetraethylene glycol

863.2 (5 mol) tetraethylene glycol are dissolved in 2.5 l of anhydrous toluene and at room temperature in succession with 159.4 g (2.5 mol) of 88% potassium hydroxide powder and 7.5 g of tetrabutylammonium hydrogen sulfate were added. At 0 ° C a solution of 53 ml (500 mmol) of 4-fluorine is then stirred vigorously Nitrobenzene in 500 ml of anhydrous toluene added dropwise. After finish The addition is after 4 hours at 0 ° C and 12 hours at room temperature stirred. The reaction mixture is worked up in 2000 ml of ice water stirred in and then the toluene phase is separated and the aqueous Phase exhaustively extracted with toluene. After washing the united organic phases with water is dried over sodium sulfate, filtered and the solvent is removed to dryness in vacuo. The remaining one a slightly yellowish, oily residue is purified by column chromatography nigt (eluent: ethyl acetate / hexane = 3: 1). 135.8 g (86.1% of theory) of one are obtained light yellowish oil.  

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 (purchased from Aldrich) is dissolved in 40 ml of absolute 1,2- Dissolved dichloroethane and with a solution of 8.0 g (25.6 mmol) of the title compound tion from Example 35a) in 15 ml of 1,2-dichloroethane. Then be 3.9 ml (33.3 mmol) of tin tetrachloride were slowly added dropwise at room temperature and the reaction solution thus obtained is left for 12 hours at room temperature stirred. For working up, water is added, the organic phase separated and the remaining aqueous phase exhaustively with dichloro extracted methane. After drying the organic phases over sodium sulfate is filtered and the solvent removed in vacuo to dryness The remaining oily residue is by means of column chromatography purified (eluent: dichloromethanol / methanol = 20: 1). 14.6 g are obtained (88.2% of theory) of 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-galactopyranosyl-1-O - [(4-nitrophenyl) tetraethylene glycol]

33.93 g (52.56 mmol) of the title compound from Example 35b) are dissolved in 680 ml of absolute methanol and mixed with 0.8 g of sodium methoxide. 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 removed in vacuo to dryness. 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 35c) is carried out according to the procedures described in the literature (see: 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 catalyst and anhydrous methanol as Lö means. After working up, 13.15 g (96% of theory) of the title compound are obtained binding as a colorless oil.

Elemental analysis (calculated on anhydrous substance):
calc .: C 53.04; H 6.57; S 6.74;
Found: C 53.13; H 6.60; S 6.84;

f) 24-cascade amine (Gd-Gly Me-DOTA amide) ₁₀ - [1-O- (4-thioureido tetraethylene glycolyl-pheny) -β-D-galactopyranosyl] ₁₄ conjugate

In an analogous manner as described in Example 1b), the conversion of 4.0 g (0.448 mmol of the title compound from Example 31a) corresponding to 6.28 mmol of free amine functions) with 14.93 g (31.4 mmol) of the title compound from example 35e) after working up 6.01 g (86.3% of theory, based on the set polyamine) as a colorless lyophilisate.

Elemental analysis (calculated on anhydrous substance):
calc .: C 47.36; H 6.28; N 8.56; S 2.89; Gd 10.11;
Found: C 47.41; H 6.31; N 8.49; S 2.93; Gd 10.18;

The ninhydrin reaction as well as the TNBS method are negative, i. H. the polymeric product contains no free amino functions. From the molar Sulfur value of the elemental analysis of the title compound gives a Bela degree of polymer with 58.3% D-galactopyranose (corresponding to 14  Galactose residues per molecule) and 41.7% Gd-Gly-DOTA (corresponding to 10 Gd-Gly-Me-DOTA complexes per molecule).

Example 36 24-cascade amine (Gd-Gly Me-DOTA amide) ₁₀ - [1-O- (4-thioureido tetraethylene glycolyl-pheny) -α-D-mannopyranosyl] ₁₄ conjugate a) α-D-Mannopyrannosyl-1-O - [(4-isothiocyanatophenyl) tetraethylene glycol]

In an analogous manner, as in Examples 35b) -35e) for the synthesis of the Ti described from Example 35e), the reaction of 15 g of α- D-mannose pentaacetate (38.4 mmol, purchased from Sigma) to form 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 53.04; H 6.57; S 6.74;
Found: C 53.14; H 6.60; S 6.68;

b) 24-cascade amine (Gd-Gly Me-DOTA amide) ₁₀ - [1-O- (4-thioureido tetraethylene glycolyl-pheny) -α-D-mannopyranosyl] ₁₄ conjugate

By carrying out the reaction analogously, as described in Example 1b), the Reaction of 5.0 g (0.56 mmol, corresponding to 7.85 mmol of free amino functions, the title compound from Example 31a) with 18.66 g (39.25 mmol) of Title compound from Example 36a) based on the formation of 7.5 g (86.3% of theory) on polyamine of the above title compound used as a colorless Lyophilisate.

Elemental analysis (calculated on anhydrous substance):
calc .: C 47.36; H 6.28; N 8.56; S 2.89; Gd 10.11;
Found: C 47.31; H 6.36; N 8.49; S 2.94; Gd 10.22;

The ninhydrin reaction as well as the TNBS method are negative, i. H. the polymeric product contains no free amino functions. From the molar  Sulfur value of the elemental analysis of the title compound gives a Bela degree of polymer expansion with 58.8% D-mannopyranose (corresponding to 14 Man residues per molecule) and 41.2% Gd-Gly-DOTA (corresponding to 10 Gd-Gly- Me-DOTA complexes per molecule).

Example 37 24-cascade amine (Gd-Gly Me-DOTA amide) ₁₀ - [1-O- (4-thioureido tetraethylene glycolyl-pheny) -N-acetyl-β-D-glucosaminopyranosyl] ₁₄ conjugate a) 1-O- [4-isothiocyanatophenyl] tetraetylene glycolyl-N-acetyl-β-D- glucosaminopyranose

In a manner analogous to that described in Examples 35b-35e), Um Putting 10 g (25.68 mmol) of N-acetyl-β-D-glucosamine pentaacetate into the bil of 9.85 g (74.3% of theory over 4 synthesis steps) of the above Title compound as a colorless oil.

Elemental analysis (calculated on anhydrous substance):
calc .: C 53.48; H 6.63; N 2.71; S 6.21;
Found: C 53.60; H 6.59; N 2.74; S 6.28;

b) 24-cascade amine (Gd-Gly Me-DOTA amide) ₁₀ - [1-O- (4-thioureido tetraethylene glycolyl-pheny) -N-acetyl-β-D-glucosaminopyranosyl] ₁₄ conjugate

By carrying out the reaction analogously to that described in Example 1b), the Reaction of 4.0 g (0.45 mmol, corresponding to 6.3 mmol of free amine radio ions) of the title compound from Example 31a) with 16.2 g (31.5 mmol) of the title Compound from Example 37a) to form 6.15 g (85% of theory, based on used polyamine) of the above-mentioned title compound as a colorless lyo philisat.

Elemental analysis (calculated on anhydrous substance):
calc .: C 47.75; H 6.32; N 9.47; S 2.78; Gd 9.75;
Found: C 47.84; H 6.38; N 9.59; S 2.86; Gd 9.80;

Example 38 24-cascade amine (Gd-Gly Me-DOTA amide) ₁₀ - [1-O- (4-thioureido tetraethylene glycolyl-pheny) -β-D-glucopyranosyl] ₁₄ conjugate a) 1-O- [4-isothiocyanatophenyl] tetraetylene glycolyl-β-D-glucopyranose

In a manner analogous to that described in Examples 35b-35e), Um setting of 8.52 g (25.68 mmol) of β-D-glucopyranosyl tetraacetate (representation according to: Rosemann S., et al., Biochemistry, Vol. 8, No. 4, 1969, 1351-1359; Pri har, Behrmann, Biochemistry 1973, 12, 997-1000) to form 9.92 g (76.8 % d. Th. Over 4 synthesis steps) of the above title compound as color loose oil (for analog synthesis see Ref. Monsigny, M., et al., Biol. Cell., 51, 187 (1984)).

Elemental analysis (calculated on anhydrous substance):
calc .: C 50.09; H 5.80; N 2.78; S 6.37;
Found: C 50.14; H 5.86; N 2.86; S 6.40;

b) 24-cascade amine (Gd-Gly Me-DOTA amide) ₁₀ - [1-O- (4-thioureido tetraethylene glycolyl-pheny) -β-D-glucopyranosyl] ₁₄ conjugate

The reaction is carried out analogously as described in Example 1b) Reaction of 4.0 g (0.45 mmol, corresponding to 6.3 mmol of free amine functions) of the title compound from Example 31a) with 15.86 g (31.5 mmol) of Title compound from Example 38a) to form 6.04 g (84.2% of theory, bezo gene on polyamine used) of the above-mentioned title compound as colorless this 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 46.28; H 5.91; N 9.63; S 2.76; Gd 9.80;

Example 39 12-cascade polyamine ( ¹¹¹ In-DTPA amide) ₄ - [β-1-O- (6-thioureidohexyl) sialyl-Le ^x ] ₈ conjugate a) 12-cascade polyamine (DTPA amide) ₄ conjugate

A solution of 5.0 g (4.07 mmol; corresponding to 48.8 mmol of free amino functions) 12-part cascade polyamine (preparation according to: Schmitt-Willich et al .; EP 0271180) is dissolved in 400 ml of distilled water and with 1 molar Sodium hydroxide adjusted a pH of 9.5. A total of 14.7 g (36.5 mmol) of DTPA monoanhydride monoethyl ester is then added, 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, the volume is made up to 800 ml with distilled water. The aqueous product solution thus obtained is ultrafiltered three times using 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: 11.2 g (89.3% of theory; based on the polyamine used) as an amorphous powder
Water content: 9.41%

Elemental analysis (calculated on anhydrous substance):
calc .: C 43.99; H 5.78; N 14.98; Na 11.92;
Found: C 44.08; H 5.82; N 14.90; Na 11.87;

After elemental analysis as well as colorimetric titration with 0.0025 molar Gadolinium sulfate standard solution in the presence of xylenol orange as an indicator, this results in a DTPA degree of loading of the polymer of 32.94% (corresponding 4 DTPA units per molecule). After quantitative ninhydrin reaction as well quantitative amino group determination using the TNBS method (cf.  Fields, R., J. Biochem. 1971, 124, 581-590) are on average per Polymer molecule 13 free amino functions before.

b) β-1-O- [6-isothiocyanatohexyl] sialyl Le ^x glycoside

150 mg (0.146 mmol) of β-1-O- [6-aminohexyl] sialyl-Le ^x glycoside (representation according to: Kunz, H. et al., Angew. Chem. 1994, 106, No. 20; Kretzschmar et al., Tetrahedron, Vol. 51, No. 47, 13015-13030, 1995) are dissolved in 20 ml of 80% aqueous ethanol, and 0.028 ml (0.365 mmol) of thiophosgene are added at 0 ° C. by addition via a microsyringe. After a reaction time of 1 hour at 22 ° C, the ethanol is drawn off in vacuo and the aqueous residue of the crude isothiocyanate obtained is used directly for the subsequent reaction, the content of the above-mentioned title compound in the aqueous crude product solution thus obtained being 0.138 mmol (corresponding to a 95 % sales) is assumed. Thin layer chromatography (ethyl acetate / methanol / water / glacial acetic acid = 5: 3: 3: 0.5; R _f = 0.72) shows a uniform reaction to the title compound mentioned above.

Elemental analysis (sulfur determination; based on anhydrous substance):
calculated: S 3.00;
found: S 2.92;

c) 12-cascade polyamine (DTPA amide) ₄ - [β-1-O- (6-thioureidohexyl) sialyl-Le ^x ] ₈ conjugate

The reaction of 26.6 mg (0.00842 mmol; corresponding to 0.0673 mmol of free amino functions) of the title compound from Example 39a) with the entire aqueous solution of the crude product from Example 39b) (0.138 mmol) yields 88 after working up and freeze drying. 7 mg (90.6% of theory) of an amorphous and colorless solid.
Water content: 8.74%

Elemental analysis (determination of sodium and sulfur; based on anhydrous substance):
calc .: S 2.20; Na 3.16;
found: S 2.17; Na 3.24;

The ninhydrin reaction as well as the TNBS determination of the title compound are negative, ie there are no free amino functions in the polymer. The percentage sulfur value of the elemental analysis of the title compound gives a degree of loading of the polymer with 67.4% sialyl-Le ^x units (corresponding to 8 sialyl-Le ^x residues per molecule).

From quantitative sialic acid determination using TBA (thiobarbituric acid) using a procedure known from the literature [cf. Lit .: Aminoff, D. (1961); "Methods for the quantitative estimation of N-acetylneuraminic acid and their application to hydrolysates of sialomucoids", J. Biochem., 81: 384-392; Uschida, Y. and Su gimori, T. (1977); "Distribution of neuraminidase in Arthrobacter and its purification by affinity chromatography", J. Biochem. (Tokyo), 82, 1425-1433] the degree of loading of the abovementioned title compound with 69.2% of tetrasaccharide (corresponding to 8 sialyl-Le ^x residues per molecule) is determined. After another colorimetric titration of the title compound with 0.0025 molar gadoliniumsulfate standard solution, in the presence of xylenol orange as an indicator, the degree of DTPA loading of the polymer of 32.9% (corresponding to 4 DTPA units per molecule) was confirmed.

d) 12-cascade polyamine ( ¹¹¹ In-DTPA amide) ₄ - [β-1-O- (6-thioureidohexyl) sialyl-Le ^x ] ₈ conjugate

1 mg of the title compound from Example 39c) are dissolved in 2 ml of sodium citrate buffer (pH 5.3) and a solution of ¹¹¹ indium chloride, which has an initial activity of 20.1 MBq, is added at room temperature. After a stirring time of 15 minutes at room temperature, the reaction solution is adjusted to pH 7.2 with 1 molar sodium hydroxide solution and then sterile filtered. The solution thus prepared can be used directly for the corresponding radio diagnostic purposes.

Example 40 12-cascade polyamine ( ¹¹¹ In-DTPA amide) ₄ - [1- (4-thioureidophenyl) -α- D-mannopyranosyl] ₈ conjugate a) 12-cascade polyamine (DTPA amide) ₄ - [1- (4-thioureidophenyl) -α- D-mannopyranosyl] ₈ conjugate

In an analogous manner to that for the synthesis of the title compound from Example 1b) described, provides the reaction of 0.5 g (0.16 mmol; corresponding to 1.3 mmol of free amine functions) of the title compound from Example 39a) with 2.07 g (6.63 mmol) p-isothio-cyanatophenyl-α-D-mannopyranose after working up tion and freeze-drying 794 mg (88.7% of theory) of the title compound as color loose and amorphous powder with a water content of 8.31%.

Elemental analysis (calculated on anhydrous substance):
calc .: C 46.61; H 5.35; N 10.27; S 4.59; Na 6.58;
Found: C 46.57; H 5.41; N 10.30; S 4.63; Na 6.52;

Both the ninhydrin reaction and the TNBS determination of the title compound dung are negative, d. H. there are no free amino functions in the polymer more before. From the percentage sulfur value of the elementary analysis of the titles connection results in a degree of loading of the polymer with 67.3% D-mannose (corresponding to 8 mannose residues per molecule). The colorimetric phenolic Determination of sulfuric acid to determine the mean pyranose load degree of conjugate gives a loading level of 8 α-D- Mannose residues per molecule and is therefore identical to the degree of loading (8th D-mannose residues per molecule), which is determined by the percentage sulfur the elementary analysis was determined. With the degree of loading determined in this way the title compound from Example 39a) of 32.94% thus results in a stati tical, average total loading of the polymer of 4-DTPA- and 8-α-D-Man residue residues per molecule.  

b) 12-cascade polyamine ( ¹¹¹ In-DTPA amide) ₄ - [1- (4-thioureioureidophenyl) -α- D-mannopyranosyl] ₈ conjugate

In an analogous manner to that described for the synthesis of the title compound from Example 39d), the title compound from Example 40a) is complexed with ^111- indium chloride (as a solution with a specific initial activity of 19.83 MBq). After sterile filtration, a preparation is obtained which can be used directly for the corresponding radio diagnostic purposes.

Example 41 24-cascade amine (Gd-Gly-Me-DOTA amide) ₁₁ - [(β-D-galactopyranosyl) -1-thio-2-imino-2-amidine] ₁₃ conjugate a) 24-cascade amine (Gly-Me-DOTA amide) ₁₁ conjugate

2.0 g (0.22 mmol) of the title compound from Example 10a) are dissolved in 60 ml of absolute dichloromethane. A total of 75 ml of tri fluoroacetic 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 drawn off to dryness in vacuo, then dissolved in 200 ml of distilled water and the aqueous product solution extracted twice with 60 ml of diethyl ether each time. The aqueous product solution thus obtained is made up to a total volume of 300 ml by adding water and ultra-filtered three times using a YM3 ultrafiltration membrane (AMICON®) against distilled water. The remaining residue is made up to a volume of 300 ml with deionized water and a pH of 10.0 is established by adding 15% sodium hydroxide solution. The basic product solution is ultrafiltered again twice and the remaining residue, after filling up with distilled water to a total volume of 500 ml, freeze-dried. 1.42 g (82.8% of theory) of the title compound are obtained as amorphous lyophilisate in the form of the free polyamine.
Water content: 5.37%

Elemental analysis (calculated on anhydrous substance):
calc .: C 51.94; H 7.83; N 17.92; S 3.77; Na 4.32;
Found: C 51.89; H 7.80; N 18.01; S 3.69; Na 4.28;

According to elemental analysis, the title compound is free of fluoride ions.

After a quantitative ninhydrin reaction, there are an average of 13 free amino functions in the title link. After colorimetric titration of the title compound with 0.0025 molar gadolinium sulfate standard solution at 60 ° C, in the presence of Xylenol orange as an indicator, there is a degree of loading of the polymer with Gly-Me-DOTA of 46.99999 00070 552 001000280000000200012000285919988800040 0002019728954 00004 998803% (corresponding to 11 Gly-Me-DOTA units) per month lekül.

b) 24-cascade amine (Gly-Me-DOTA amide) ₁₁ - [(β-D-galactopyranosyl) - 1-thio-2-imino-2-amidine] ₁₃ conjugate

In an analogous manner, as in the literature for the reactions with proteins (see: Lee, Y.C. et al., Methods of Enzymology, Vol. 83, 1982, 278) provides the reaction of 1.0 g (0.128 mmol; corresponding to 1.66 mmol of free amine functions) of the title compound from Example 41a) with 11.4 g (42.8 mmol) 2-imino-2-methoxyethyl-1-thio-β-D-galactopyranose (IME-thio galactoside) 1.06 g (76.3% of theory) of the title compound as a colorless lyophilisate [ For the synthesis and the reaction conditions of analogous coupling reactions of IME thiopyranosides on proteins see: Y. C. Lee, C. P. Stowell, M. J. Krantz, Biochemistry 1976, 15, 3956; R. Roger, D.G. Neilson, Chem. Rev. 1961, 61, 179; L. Wofsy, S.J. Singer, Biochemistry 1963, 2,104; M. J. Hunter, M. L. Lud wig, this series, vol. 25, 585; J.E. Zull, J. Chung, J. Biol. Chem. 1975, 250, 1668; C. P. Stowell, Doctoral Dissertation, The Johns Hopkins University, Balti more, Maryland, 1978].

Elemental analysis:
calc .: C 57.67; H 8.03; N 9.73; S 2.90;
Found: C 57.78; H 8.14; N 9.80; S 2.97;

The title compound shows a negative ninhydrin reaction. From the percent len sulfur value of the elementary analysis results in an average degree of loading of the polymer with 13 D-galactose residues per molecule.  

c) 24-cascade amine (Gd-Gly-Me-DOTA amide) ₁₁ - [(β-D-galactopyra nosyl) -1-thio-2-imino-2-amidine] ₁₃ conjugate

In an analogous manner as described for Example 23d), the complexation of 900 mg (0.082 mmol; corresponding to 0.90 mmol of Gly-Me-DOTA) of the title compound from Example 41b) with 241 mg (0.92 mmol) of gadolinium chloride leads to dissolved in 10 ml of distilled water to form the above title compound.
Yield: 990 mg (96.0% of theory) as a colorless and amorphous powder
Water content: 9.32%

Elemental analysis (calculated on anhydrous substance):
calc .: C 42.19; H 6.06; N 12.58; S 3.31; Gd 13.75;
Found: C 42.23; H 6.10; N 12.57; S 3.28; Gd 13.80;

The average degree of loading of the polymer of the above title compound was made from the molar element ratios of sulfur to gadolinium 11 Gd-Gly-Me-DOTA units and 13 β-D-glucopyranosyl residues determined, what about the loading levels of the title compound determined on the preliminary stages correlation very well.

Example 42 24-cascade amine (Yb-DTPA amide) ₁₁ - [1- (4-thioureidophenyl) -β-D-galacto pyranosyl] ₁₃ conjugate

1.0 g (0.077 mmol; corresponding to 0.85 mmol of DTPA) of the title compound from Example 4b) are dissolved in 40 ml of sodium citrate buffer (pH 5.3) and at room temperature with a solution of 243 mg (0.87 mmol) Ytterbium 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: 969 mg (96.2% of theory) as an amorphous powder
Water content: 4.93%

Elemental analysis (calculated on anhydrous substance):
calc .: C 41.48; H 4.91; N 9.74; S 3.19; Yb 14.54; Na 1.93;
Found: C 41.52; H 4.96; N 9.80; S 3.22; Yb 14.60; Na 1.88;

The ninhydrin reaction as well as the TNBS determination of the title compound ver run negative, d. H. there are no longer any free amino functions in the polymer. From the percentage sulfur value of the elemental analysis of the title compound the degree of loading of the polymer is 55.2% D-galactose (corresponding to 13 galactose residues per molecule) while from the percentage Ytterbium Value a degree of loading of the polymer with ytterbium DTPA of 44.8 % results (corresponding to 11 Yb-DTPA units per molecule). The colorimetri phenol-sulfuric acid determination [Dubois et al., Anal. Chem. 1956, 28, 3, 350; Koch et al., Cereal Chem. 1951, 28, 424] as well as the quantitative Carbohydrate determination method according to Nelson and Somogyi [Cheronis et al., Microchim. Acta, 6, 769 (1957); Somogyi, J. Biol. Chem. 195, 19 (1952); Nel son, J. Biol. Chem., 153, 375 (1944); see. also Hodge et al., Meth. Carbohydra te Chem., Vol. 1 ,; Whistler and Wolfrom, eds. p. 388, Academic Press, New York, 1962; So and Goldstein, J. Biol. Chem. 242, 1617 (1967)] for the determination the average pyranose loading level of the polyamine provides egg as a result degree of loading of 32.68 D-galactose residues per molecule and is thus identical to the degree of loading, which is determined by the percentage element was determined.

Example 43 24-cascade amine (Dy-DTPA amide) ₁₁ - [1- (4-thioureidophenyl) -α- D-mannopyranosyl] ₁₃ conjugate

1.4 g (0.108 mmol; corresponding to 1.18 mmol of DTPA) of the title compound from Example 5a) are dissolved in 40 ml of sodium citrate buffer (pH 5.3) and at room temperature with a solution of 325 mg (1.21 mmol) Dysprosium 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: 1.35 g (97.0% of theory) as an amorphous powder
Water content: 6.74%

Elemental analysis (calculated on anhydrous substance):
calc .: C 41.86; H 4.95; N 9.83; S 3.21; Dy 13.78; Na 1.95;
Found: C 41.84; H 4.99; N 9.78; S 3.25; Dy 13.81; Na 1.89;

From the molar elementary ratio of dysprosium to sulfur thus an average degree of loading of the polymer of 46.2% Gd-DPTA and 53.8% D-mannose (corresponding to 11 Dy-DTPA units and 13 D-mannose residues per molecule).

The qualitative ninhydrin reaction as well as the TNBS determination of the title ver bond run negative, d. H. there are no free amino functions in the polymer more before. The colorimetric phenol-sulfuric acid determination, as well as the quantitative carbohydrate determination method according to Nelson and Somogyi for Determination of the average pyranose loading level of the polyamine delivers as The result is a degree of loading of 32.68 D-mannose residues per molecule thus identical to the degree of loading, which is determined by the percentage el was determined.

Example 44 24-cascade amine (Mn-DTPA amide) ₁₁ - [1- (4-thioureidophenyl) -N-acetyl-β-D-glucosaminopyranosyl] ₁₃ conjugate

1.0 g (0.077 mmol; corresponding to 0.85 mmol of DTPA) of the title compound from Example 4b) are dissolved in 40 ml of sodium citrate buffer (pH 5.3) and at room temperature with 100 mg (0.87 mmol) of manganese-II -carbonate added. 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: 911 mg (98.3% of theory) as an amorphous powder
Water content: 7.46%

Elemental analysis (calculated on anhydrous substance):
calc .: C 45.09; H 5.33; N 10.59; S 3.46; Mn 5.02; Na 4.20;
Found: C 45.11; H 5.38; N 10.61; S 3.42; Mn 5.10; Na 4.13;

The ninhydrin reaction as well as the TNBS determination of the title compound ver run negative, d. H. there are no longer any free amino functions in the polymer. From the percentage sulfur value of the elemental analysis of the title compound there is a degree of loading of the polymer with 53.8% N-acetylglucosamine (equivalent to 13 N-acetyl-β-D-glucosamine residues per molecule) while from the percentage of manganese a degree of loading of the polymer with manganese DTPA of 46.0% results (corresponding to 11 Mn DTPA units per mole cool). The colorimetric phenol-sulfuric acid determination, as well as the quantitative carbohydrate determination method according to Nelson and Somogyi for Determination of the average pyranose loading level of the polyamine, delivers as Result a loading level of 13 N-acetyl-β-D-glucosamine residues per mo lekül and is therefore identical to the degree of loading, which is determined by the pro elemental relationships were determined.

Example 45 24-cascade amine (Eu-DTPA amide) ₁₁ - [1- (4-thioureidophenyl) -β-D-gluco pyranosyl] ₁₃ conjugate

1.4 g (0.108 mmol; corresponding to 1.18 mmol of DTPA) of the title compound from Example 7b) are dissolved in 40 ml of sodium citrate buffer (pH 5.3) and at room temperature with a solution of 312 mg (1.21 mmol) Europium 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: 1.30 g (97.6% of theory) as an amorphous powder
Water content: 8.34%

Elemental analysis (calculated on anhydrous substance):
calc .: C 42.23; H 4.99; N 9.92; S 3.24; Eu 13.00; Na 1.97;
Found: C 42.24; H 5.03; N 9.89; S 3.19; Eu 13.07; Na 1.93;

The molar element ratio of europium to sulfur results in thus an average degree of loading of the polymer of 45% Eu-DTPA units and 55% β-D-glucose residues (corresponding to 11 DTPA units and 13 β-D- Glucose units per molecule). In the title compound are using ninhydrin reaction no free amino functions detectable.

The qualitative ninhydrin reaction as well as the TNBS determination of the title ver bond run negative, d. H. there are no free amino functions in the polymer more before. The colorimetric phenol-sulfuric acid determination, as well as the quantitative carbohydrate determination method according to Nelson and Somogyi for Determination of the mean pyranose loading level of the conjugate delivers as The result is an average loading of 13 D-glucose residues per molecule and is therefore identical to the degree of loading, which is determined by the percentage Element ratios was determined.

Example 46 Amide conjugate of 24-polyamine based on the N, N, N ', N', N ", N" - Hexakis [2- (trilysyl-amino) -ethyl] trimesic acid triamide with the Gadolinium complex of 10- (4-carboxy-1-methyl-2-oxo-3-azabutyl) -1,4,7, 10-tetraazacyclododecane-1,4,7-triacetic acid and 1-carboxymethyl-β-D- glucopyranoside (m = 13, n = 11) a) 1-Tert.butoxycarbonylmethyl-2,3,4,6-tetra-O-benzyl-β-D-glucopyranoside

To a stirred solution of 661 mg (5.0 mmol) of tert-butyl glycolate in 25 ml of absolute 1,2-dichloroethane are added in succession at room temperature the finely powdered molecular sieve 4 O (5.0 g) and silver silicate (5.0 g). On finally, dropwise at 0 ° C with a solution of 3.02 g (5.0 mmol) 2,3,4,6-tetra-O-benzyl-α-D-glucopyranosyl bromide, dissolved in 40 ml solute 1,2-dichloroethane. The reaction solution thus obtained is on finally stirred at -10 ° C for 3 hours. The mixture is worked up at 0 ° C mixed with 250 ml of 1,2-dichloroethane and the reaction solution on Celite sucked. The filtrate is washed in succession with 200 ml of aqueous sodium hydro gene carbonate solution and 220 ml of water, washed over sodium sulfate  dries and filtered. After removing the solvent in vacuo the remaining residue is chromatographed on silica gel (eluent: toluene: Ethyl acetate = 10: 1).

488.7 mg (15.3% of theory) of the above-mentioned title compound are obtained, as a colorless oil.

Elemental analysis:
calc .: C 73.37; H 7.08;
Found: C 73.21; H 7.24;

b) 1-carboxymethyl-2,3,4,6-tetra-O-benzyl-β-D-glucopyranoside

50 ml of trifluoroacetic acid are added to 30 g (45.81 mmol) of the title compound from Example 46a) dissolved in 300 ml of 1,2-dichloroethane at -10 ° C. and the mixture is stirred for 2 hours at this temperature. The mixture is then stirred at 0 ° C for one hour. It is evaporated to the dry state in a vacuum and the residue is chromatographed on silica gel (mobile solvent: n-hexane / 2-propanol = 5: 1).
Yield: 3.02 g (11.4% of theory) of a glassy solid.

Elemental analysis:
calc .: C 72.22; H 6.40;
Found: C 72.09; H 6.61;

c) Amide conjugate of the 24-polyamine based on the N, N, N ', N', N ", N" - Hexakis [2- (trilysyl-amino) -ethyl] trimesic acid triamide with the Gadolinium complex of 10- (4-carboxy-1-methyl-2-oxo-3-azabutyl) - 1,4,7,10-tetraazacyclododecane-1,4,7-triacetic acid and 1- Carboxymethyl-2,3,4,6-tetra-O-benzyl-β-D-glucopyranoside (m = 13, n = 11)

12.28 g (19.5 mmol) of the Gd complex of 10- (4-carboxy-1-methyl-2-oxo-3-azabutyl) -1,4,7,10-tetraazacyclododecane-1 described in Example 31h), 4,7-triacetic acid, 2.0 g sodium bromide (19.5 mmol), 8.28 g (72 mmol) N-hydroxy succinimide and 9.88 g (16.5 mmol) the title compound from Example 46b) who in 200 ml of dimethyl sulfoxide 50 ° C solved. After cooling to room temperature, 14.86 g (72 mmol) of N, N'-dicyclohexylcarbodiimide are added and the mixture is preactivated for 60 minutes. A solution of 1.39 g (0.5 mmol) of the 24-mer polyamine based on the N, N, N ', N', N ", N" - is added to the N-hydroxysuccinimide ester solution thus prepared. Hexakis [2. (trilysylamino) ethyl] tri mesinsäuretriamids and 7.28 g (72 mmol) triethylamine in 10 ml of water and stirred overnight at room temperature. The suspension obtained is filtered and then mixed with sufficient acetone until precipitation is complete, the precipitate is filtered off with suction, dried, taken up in water, filtered off from the insoluble dicyclohexylurea and the filtrate is desalted and cut through an AMICON® YM-3 ultrafiltration membrane (cut off 3000 Da) cleaned of low-molecular components. The retentate is then freeze-dried.
Yield: 8.13 g (95% of theory)
H ₂ O content (Karl Fischer): 7.8%

Elemental analysis (calculated on anhydrous substance):
calc .: C 54.19; H 5.98; N 9.00; Gd 11.95;
found: C 54.06; H 6.10; N 8.87; Gd 11.81;

A ratio of m: n = 13:11 can be calculated from this.

d) amide conjugate of the 24-polyamine based on the N, N, N ', N', N ", N" - Hexakis [2- (trilysyl-amino) -ethyl] trimesic acid triamide with the Gadolinium complex of 10- (4-carboxy-1-methyl-2-oxo-3-azabutyl) - 1,4,7,10-tetraazacyclododecane-1,4,7-triacetic acid and 1-carboxy methyl-β-D-glucopyranoside (m = 13, n = 11)

8.0 g (0.467 mmol) of the title compound from Example 46c) are dissolved in a mixture of 50 ml. Ethanol / 100 ml water and 2 g palladium catalyst (10% Pd on activated carbon) are added. It is hydrogenated for 12 hours at room temperature. The catalyst is filtered off and the filtrate is evaporated to dryness in vacuo. The residue is dissolved in 300 ml of water and freeze-dried.
Yield: 6.08 g (99% of theory) of a glassy solid.
H ₂ O content (Karl Fischer): 11%

Elemental analysis (calculated on anhydrous substance):
calc .: C 42.39; H 5.76; N 11.72; Gd 15.55;
Found: C 42.21; H 5.89; N 11.61; Gd 15.42;

A ratio of m: n = 13:11 can be calculated from this.

Example 47 Thiourea conjugate of 24-polyamine based on the N, N, N ', N', N ", N" -Hexakis [2- (trilysylamino) ethyl] trimesic acid triamide with the gadolinium complex based on the 4-isothiocyanatobenzyl-DOTA and 1-carboxymethyl-α-D-mannopyranoside (m = 10, n = 14) a) Thiourea conjugate of 24mer polyamine based on N, N, N ', N', N ", N" -Hexakis [2- (trilysylamino) ethyl] trimesic acid triamide with 1-carboxymethyl-2,3,4,6-tetra-O-benzyl-α-D-mannopyranoside (m = 10, n = 14).

Dissolved to 1.39 g (0.5 mmol) of the 24mer polyamine based on the N, N, N ', N', N ", N" - hexakis (2- (trilysylamino) ethyl) trimesic acid triamide 5.46 g (54 mmol) of triethylamine are added to 50 ml of water. A mixture of 4.11 g (7.5 mmol) of 4-isothiocyanatobenzyl-DOTA (US Pat. No. 4,923,985; Gansow OA et al., J. Chem. Soc. Perkin Trans. 1, 1992, 1173) is added dropwise to this solution at 10 ° C. - 1178) and 3.28 g (10.5 mmol) of p-isothiocyanatophenyl-α-D-mannopyranose, dissolved in 75 ml of water / 50 ml of ethanol (dropping time: 2 hours). The mixture is stirred at room temperature for 12 hours. Then add 3.54 g (10.6 mmol) of gadolinium acetate and adjust to pH 4.5 with 10% hydrochloric acid. The mixture is stirred for 5 hours at 80 ° C., allowed to come to room temperature and adjusted to pH 7.5 with 5% aqueous NaOH. The solution is desalted using an AMICON® YM-3 Ultra filtration membrane (cut off 3000 Da) and cleaned of low molecular weight components. The retentate is then freeze-dried.
Yield: 6.76 g (94% of theory) of a glassy solid.
H ₂ O content (Karl Fischer): 9.8%

Elemental analysis: (calculated on anhydrous substance):
calc .: C 46.01; H 4.91; N 10.61; Gd 10.93; S 5.35; Na 1.60;
Found: C 45.87; H 4.99; N 10.50; Gd 10.82; S 5.21; Na 1.42;

A ratio of m: n = 10: 14 can be calculated from this.

Example 48 Thiourea conjugate of 24-polyamine based on the N, N, N ', N', N ", N" - Hexakis [2- (trilysyl-amino) -ethyl] trimesic acid triamide with the Gadolinium complex based on 4-isothiocyanatobenzyl-DTPA and 1-carboxymethyl-β-D-galactopyranoside (m = 8, n = 16) a) Thiourea conjugate of 24mer polyamine based on the N, N, N ', N', N ", N" -Hexakis [2- (trilysylamino) ethyl] trimesic acid triamide with 4-isothiocyanatobenzyl-DTPA (n = 8, n = 16)

To 1.39 g (0.5 mmol) of the 24mer-polyamine based on the N, N, N ', N', N ", N" - hexakis [2- (trilysylamino) ethyl] -trimesic acid triamide dissolved in 50 ml of water are added to 5.46 g (54 mmol) of triethylamine. A mixture of 3.22 g (6.0 mmol) of 4-isothiocyanotobenzyl-DTPA (WO 91/14459; Gansow, OA et al., Bioconjugate Chem. 1992, 3, 108-117) is added dropwise to this solution at 10 ° C. ) and 3.75 g (12.0 mmol) of p-isothiocyanatophenyl-β-D-galactopyranose, dissolved in 75 ml of water / 50 ml of ethanol (dropping time: 2 hours). The mixture is stirred at room temperature for 12 hours. Then 2.04 g (6.1 mmol) of gadolinium acetate are added and the pH is adjusted to 4.5 with 10% hydrochloric acid. The mixture is stirred at 60 ° C. for 3 hours. The mixture is allowed to come to room temperature and adjusted to pH 7.5 with 5% NaOH. The solution is desalted via an AMICON® YM-3 ultrafiltration membrane (cut off 3000 Da) and cleaned of low molecular weight components. The residue is then freeze-dried.
Yield: 6.55 g (96% of theory) of a glassy solid.
H ₂ O content (Karl Fischer): 10%

Elemental analysis: (calculated on anhydrous substance):
calc .: C 45.15; H 4.66; N 9.54; Gd 9.22; S 5.64; Na 2.70;
Found: C 45.03; H 4.81; N 9.44; Gd 9.10; S 5.48; Na 2.51;

A ratio of m: n = 8:16 can be calculated from this.

Example 49 Amide conjugate of 24-polyamine based on the N, N, N ', N', N ", N" - Hexakis [2- (trilysyl-amino) -ethyl] trimesic acid triamide with the Gadolinium complex of 10- (4-carboxy-1-methyl-2-oxo-3-azabutyl) -1,4,7,10- tetraazacyclododecane-1,4,7-triacetic acid and 1-carboxymethyl-α-D- mannopyranoside (m = 13, n = 11) a) 1-Tert.butoxycarbonylmethyl-2,3,4,6-tetra-O-benzyl-α-D-mannopyranoside

To a stirred solution of 661 mg (5.0 mmol) of tert-butyl glycolate in 25 ml of absolute 1,2-dichloroethane are added in succession at room temperature the finely powdered molecular sieve 4 Å (5.0 g) and silver silicate (5.0 g). On finally, dropwise at 0 ° C with a solution of 3.02 g (5.0 mmol) 2,3,4,6-tetra-O-benzyl-α-D-mannopyranosyl bromide, dissolved in 40 ml absolute 1,2-dichloroethane. The reaction solution thus obtained becomes then stirred for 3 hours at -10 ° C. The mixture is worked up at 0 ° C mixed with 250 ml of 1,2-dichloroethane and the reaction solution on Celite sucked. The filtrate is washed in succession with 200 ml of aqueous sodium hydro gene carbonate solution and 220 ml of water, washed over sodium sulfate dries and filtered. After removing the solvent in vacuo the remaining residue is chromatographed on silica gel (eluent: toluene: Ethyl acetate = 10: 1).

472.7 mg (14.8% of theory) of the above-mentioned title compound are obtained, as a colorless oil.

Elemental analysis:
calc .: C 73.37; H 7.08;
Found: C 73.25; H 7.19;

b) 1-carboxycarbonylmethyl-2,3,4,6-tetra-O-benzyl-α-D-mannopyranoside

To 28 g (42.76 mmol) of the title compound from Example 49a) dissolved in 300 ml of 1,2-dichloroethane, 50 ml of trifluoroacetic acid are added at -10 ° C. and the mixture is stirred for 2 hours at this temperature. Then one hour at 0 ° C. It is evaporated to the dry state in a vacuum and the residue is chromatographed on silica gel (mobile solvent: n-hexane / 2-propanol = 5: 1).
Yield: 2.58 g (10.4% of theory) of a glassy solid.

Elemental analysis:
calc .: C 72.22; H 6.40;
found: C 72.10; H 6.58;

c) Amide conjugate of the 24-polyamine based on the N, N, N ', N', N ", N" - Hexakis [2- (trilysyl-amino) -ethyl] trimesic acid triamide with the Gadolinium complex of 10- (4-carboxy-1-methyl-2-oxo-3-azabutyl) -1,4,7, 10-tetraazacyclododecane-1,4,7-triacetic acid and 1-carboxymethyl-2,3, 4,6-tetra-O-benzyl-α-D-mannopyranoside (m = 13, n = 11)

10.39 g (16.5 mmol) of the Gd complex of 10- (4-carboxy-1-methyl-2-oxo-3-azabutyl) -1,4,7,10-tetraazacyclododecane-1 described in Example 31h), 4,7-triacetic acid, 1.7 g of sodium bromide (16.5 mmol), 8.28 g (72 mmol) of N-hydroxy succinimide and 11.67 g (19.5 mmol) of the title compound from Example 49b) who in 200 ml of dimethyl sulfoxide 50 ° C solved. After cooling to room temperature, 14.86 g (72 mmol) of N, N'-dicyclohexylcarbodiimide are added and the mixture is preactivated for 60 minutes. A solution of 1.39 g (0.5 mmol) of the 24-mer polyamine based on the N, N, N ', N', N ", N" -Hexakis [2] is added to the N-hydroxysuccinimide ester solution thus prepared (trilysylamino) ethyl] trimesin acid triamide and 7.28 g (72 mmol) triethylamine in 10 ml water and stir overnight at room temperature. The suspension obtained is filtered and then sufficient acetone is added until precipitation is complete, the precipitate is filtered off with suction, dried, taken up in water, filtered off from the insoluble dicyclohexylurea and the filtrate is desalted via an AMICON® YM-3 Ultra filtration membrane (cut off 3000 Da) and cleaned of low molecular weight components. The retentate is then freeze-dried.
Yield: 8.1 g (95% of theory)
H ₂ O content (Karl Fischer): 9.7%

Elemental analysis (calculated on anhydrous substance):
calc .: C 56.78; H 6.09; N 8.21; Gd 10.14;
Found: C 56.64; H 6.18; N 8.14; Gd 10.01;

A ratio of m: n = 11:13 can be calculated from this.

d) amide conjugate of the 24-polyamine based on the N, N, N ', N', N ", N" - Hexakis [2- (trilysyl-amino) -ethyl] trimesic acid triamide with the Gadolinium complex of 10- (4-carboxy-1-methyl-2-oxo-3-azabutyl) -1,4,7, 10-tetraazacyclododecane-1,4,7-triacetic acid and 1-carboxymethyl-α-D- mannopyranoside (m = 13, n = 11)

7.0 g (0.41 mmol) of the title compound from Example 49c) are dissolved in a mixture of 50 ml. Ethanol / 100 ml water and 2 g palladium catalyst (10% Pd on activated carbon) are added. It is hydrogenated for 12 hours at room temperature and a hydrogen pressure of 1 atm. The catalyst is filtered off and the filtrate is evaporated to dryness in vacuo. The residue is dissolved in 300 ml of water and freeze-dried.
Yield: 5.03 g (99% of theory) of a glassy solid.
H ₂ O content (Karl Fischer): 9.8%

Elemental analysis (calculated on anhydrous substance):
calc .: C 42.94; H 5.86; N 11.33; Gd 13.99;
Found: C 42.86; H 5.97; N 11.24; Gd 13.74;

A ratio of m: n = 11:13 can be calculated from this.

Example 50 Amide conjugate of 24-polyamine based on the N, N, N ', N', N ", N" - Hexakis [2- (trilysyl-amino) -ethyl] trimesic acid triamide with the Gadolinium complex of 10- (4-carboxy-1-methyl-2-oxo-3-azabutyl) -1,4,7,10- tetraazacyclododecane-1,4,7-triacetic acid and 1-carboxymethyl-β-D- galactopyranoside (m = 6, n = 18) a) 1-Tert.butoxycarbonylmethyl-2,3,4,6-tetra-O-benzyl-β-D-galactopyranoside

To a stirred solution of 661 mg (5.0 mmol) of tert-butyl glycolate in 25 ml of absolute 1,2-dichloroethane are added in succession at room temperature the finely powdered molecular sieve 4 Å (5.0 g) and silver silicate (5.0 g). On finally, dropwise at 0 ° C with a solution of 3.02 g (5.0 mmol) 2,3,4,6-tetra-O-benzyl-α-D-galactopyranosylbromide, dissolved in 40 ml absolute 1,2-dichloroethane. The reaction solution thus obtained becomes then stirred for 3 hours at -10 ° C. The mixture is worked up at 0 ° C mixed with 250 ml of 1,2-dichloroethane and the reaction solution poured off over Celite sucks. The filtrate is washed in succession with 200 ml of aqueous sodium hydrogen washed carbonate solution and 220 ml of water, dried over sodium sulfate net and filtered. After the solvent has been stripped off in vacuo, the remaining residue chromatographed on silica gel (eluent: toluene: Es ethyl acetate = 10: 1).

456.7 mg (14.3% of theory) of the above-mentioned title compound are obtained, as a colorless oil.

Elemental analysis:
calc .: C 73.37; H 7.08;
Found: C 73.18; H 7.19;

b) 1-carboxymethyl-2,3,4,6-tetra-O-benzyl-β-D-galactopyranoside

To 29 g (44.29 mmol) of the title compound from Example 50a) dissolved in 300 ml of 1,2-dichloroethane, 50 ml of trifluoroacetic acid are added at -10 ° C. and the mixture is stirred for 2 hours at this temperature. Then one hour at 0 ° C. It is evaporated to the dry state in a vacuum and the residue is chromatographed on silica gel (mobile solvent: n-hexane / 2-propanol = 5: 1.
Yield: 3.79 g (14.7% of theory) of a glassy solid.

Elemental analysis:
calc .: C 72.22; H 6.40;
found: C 72.10; H 6.51;

c) Amide conjugate of the 24-polyamine based on the N, N, N ', N', N ", N" - Hexakis [2- (trilysyl-amino) -ethyl] trimesic acid triamide with the Gadolinium complex of 10- (4-carboxy-1-methyl-2-oxo-3-azabutyl) -1,4,7, 10-tetraazacyclododecane-1,4,7-triacetic acid and 1-carboxymethyl-β-D- 2,3,4,6-tetra-O-benzyl-galactopyranoside (m = 6, n = 18)

5.67 g (9 mmol) of the Gd complex of 10- (4-carboxy-1-methyl-2-oxo-3-azabutyl) -1,4,7,10-tetraazacyclododecane-1 described in Example 31 h) , 4,7-tri acetic acid, 0.93 g of sodium bromide (9 mmol), 8.28 g (72 mmol) of N-hydroxysuccinimide and 16.16 g (27 mmol) of the title compound from Example 50b) are in 200 ml of dimethyl sulfoxide dissolved at 50 ° C. After cooling to room temperature, 14.86 g (72 mmol) of N, N'-dicyclohexylcarbodiimide are added and the mixture is preactivated for 60 minutes. A solution of 1.39 g (0.5 mmol) of the 24-mer polyamine based on the N, N, N ', N', N ", N" -Hexakis is added to the N-hydroxysuccinimide ester solution thus prepared [2. (trilysyl-amino) -ethyl] -trimesic acid triamide and 7.28 g (72 mmol) of triethylamine in 10 ml of water and stirred overnight at room temperature. The suspension obtained is filtered and then sufficient acetone is added until precipitation is complete, the precipitate is filtered off with suction, dried, taken up in water, filtered off from the insoluble dicyclohexylurea and the filtrate is desalted via an AMICON® YM-3 ultrafiltration membrane (cut off 3000 Da) and cleaned of low molecular weight constituents. The retentate is then freeze-dried.
Yield: 8.03 g (95% of theory)
H ₂ O content (Karl Fischer): 9.0%

Elemental analysis (calculated on anhydrous substance):
calc .: C 63.34; H 6.39; N 6.22; Gd 5.58;
Found: C 63.18; H 6.47; N 6.13; Gd 5.47;

A ratio of m: n = 6:18 can be calculated from this.

d) amide conjugate of the 24-polyamine based on the N, N, N ', N', N ", N" - Hexakis [2- (trilysyl-amino) -ethyl] trimesic acid triamide with the Gadolinium complex of 10- (4-carboxy-1-methyl-2-oxo-3-azabutyl) -1,4,7, 10-tetraazacyclododecane-1,4,7-triacetic acid and 1-carboxymethyl-β-D- galactopyranoside (m = 6, n = 18)

7.0 g (0.414 mmol) of the title compound from Example 50c are dissolved in a mixture of 75 ml. Ethanol / 100 ml water and 2 g palladium catalyst (10% Pd on activated carbon) are added. It is hydrogenated for 12 hours at room temperature at a hydrogen pressure of 1 atm. The catalyst is filtered off and the filtrate is evaporated to dryness in vacuo. The residue is dissolved in 300 ml of water and freeze-dried.
Yield: 4.27 g (99% of theory) of a glassy solid.
H ₂ O content (Karl Fischer): 10.3%

Elemental analysis (calculated on anhydrous substance):
calc .: C 44.67; H 6.19; N 10.09; Gd 9.07;
Found: C 44.53; H 6.28; N 9.91; Gd 8.94;

A ratio of m: n = 6:18 can be calculated from this.

Example 51 Amide conjugate of 24-polyamine based on the N, N, N ', N', N ", N" - Hexakis [2- (trilysyl-amino) -ethyl] trimesic acid triamide with the Gadolinium complex of 10- (4-carboxy-1-methyl-2-oxo-3-azabutyl) -1,4,7, 10-tetraazacyclododecane-1,4,7-triacetic acid and 1-carboxymethyl-α-L- fucopyranoside (m = 10, n = 14) a) 1-Tert.butoxycarbonylmethyl-2,3,4-tri-O-benzyl-α-L-fucopyranoside

To a stirred solution of 661 mg (5.0 mmol) of tert-butyl glycolate in 25 ml of absolute 1,2-dichloroethane are added in succession at room temperature the finely powdered molecular sieve 4 Å (5.0 g) and silver silicate (5.0 g). On finally, dropwise at 0 ° C. with a solution of 2.49 g (5.0 mmol) 2,3,4,6-tetra-O-benzyl-α-D-fucopyranosyl bromide, dissolved in 40 ml of absolute 1,2-dichloroethane. The reaction solution thus obtained is then stirred for 3 hours at -10 ° C. For working up, at 0 ° C with 250 ml 1,2- Dichloroethane was added and the reaction solution was suction filtered through Celite. The Filtrate is successively with 200 ml of aqueous sodium bicarbonate solution and 220 ml of water, dried over sodium sulfate and  filtered. After the solvent has been stripped off in vacuo, the lead becomes lead The residue was chromatographed on silica gel (eluent: toluene: acetic acid ethyl ester = 10: 1).

407.5 mg (15.3% of theory) of the above-mentioned title compound are obtained as colorless oil.

Elemental analysis:
calc .: C 72.24; H 7.35;
Found: C 72.10; H 7.45;

b) (1-carboxymethyl) -2,3,4-tri- (O-benzyl) -α-L-fucopyranoside

To 24 g (43.74 mmol) of the title compound from Example 51a) dissolved in 300 ml of 1,2-dichloroethane, 50 ml of trifluoroacetic acid are added at -10 ° C. and the mixture is stirred for 2 hours at this temperature. Then one hour at 0 ° C. It is evaporated to the dry state in a vacuum and the residue is chromatographed on silica gel (mobile solvent: n-hexane / 2-propanol = 5: 1).
Yield: 3.36 g (16.2% of theory) of a glassy solid.

Elemental analysis:
calc .: C 70.71; H 6.55;
Found: C 70.58; H 6.71;

c) Amide conjugate of the 24-polyamine based on the N, N, N ', N', N ", N" - Hexakis [2- (trilysyl-amino) -ethyl] trimesic acid triamide with the Gadolinium complex of 10- (4-carboxy-1-methyl-2-oxo-3-azabutyl) -1,4,7, 10-tetraazacyclododecane-1,4,7-triacetic acid and 1-carboxymethyl-2,3,4- tri-O-benzyl-α-L-fucopyranoside (m = 10, n = 14)

9.45 g (15 mmol) of the Gd complex of 10- (4-carboxy-1-methyl-2-oxo-3-azabutyl) -1,4,7,10-tetraazacyclododecane-1 described in Example 31 h) , 4,7-triacetic acid, 1.54 g sodium bromide (15 mmol), 8.28 g (72 mmol) N-hydroxy succinimide and 10.34 g (21 mmol) of the title compound from Example 51b) are dissolved in 200 ml dimethyl sulfoxide dissolved at 50 ° C. After cooling to room temperature, 14.86 g (72 mmol) of N, N'-dicyclohexylcarbodiimide are added and the mixture is preactivated for 60 minutes. A solution of 1.386 g (0.5 mmol) of the 24-mer polyamine based on the N, N, N ', N', N ", N" -Hexakis is added to the N-hydroxysuccinimide ester solution thus prepared [2. (trilysylamino) ethyl] trimesic acid triamids and 7.28 g (72 mmol) triethylamine in 10 ml water and stirred overnight at room temperature. The suspension obtained is filtered and then sufficient acetone is added until precipitation is complete, the precipitate is filtered off with suction, dried, taken up in water, filtered off from insoluble dicyclohexylurea and the filtrate is desalted via an AMICON® YM-3 ultra filtration membrane (cut off 3000 Da) and cleaned of low molecular weight components. The retentate is then freeze-dried.
Yield: 7.3 g (94% of theory)
H ₂ O content (Karl Fischer: 8.6%)

Elemental analysis (calculated on anhydrous substance):
calc .: C 56.06; H 6.20; N 8.57; Gd 10.12;
Found: C 55.91; H 6.32; N 8.49; Gd 9.97;

A ratio of m: n = 10: 14 can be calculated from this.

d) amide conjugate of the 24-polyamine based on the N, N, N ', N', N ", N" - Hexakis [2- (trilysyl-amino) -ethyl] trimesic acid triamide with the Gadolinium complex of 10- (4-carboxy-1-methyl-2-oxo-3-azabutyl) -1,4,7, 10-tetraazacyclododecane-1,4,7-triacetic acid and 1-carboxymethymethyl-α-L- fucopyranoside (m = 10, n = 14)

7.0 g (0.41 mmol) of the title compound from Example 51c) are dissolved in a mixture of 75 ml. Ethanol / 100 ml water and 2 g palladium catalyst (10% Pd on activated carbon) are added. It is hydrogenated for 12 hours at room temperature and a hydrogen pressure of 1 atm. The catalyst is filtered off and the filtrate is evaporated to dryness in vacuo. The residue is dissolved in 300 ml of water and freeze-dried.
Yield: 5.24 g (99% of theory) of a glassy solid.
H ₂ O content (Karl Fischer): 11.1%

Elemental analysis (calculated on anhydrous substance):
calc .: C 44.06; H 6.03; N 11.33; Gd 13.38;
Found: C 43.95; H 6.17; N 11.24; Gd 13.29;

A ratio of m: n = 10: 14 can be calculated from this.

Example 52 Amide conjugate of 24-polyamine based on the N, N, N ', N', N ", N" - Hexakis [2- (trilysyl-amino) -ethyl] trimesic acid triamide with the Gadolinium complex of 10- (4-carboxy-1-methyl-2-oxo-3-azabutyl) -1,4,7, 10-tetraazacyclododecane-1,4,7-triacetic acid and 6-carboxyhexyl-β-D- galactopyranoside (m = 10, n = 14) a) 6-Tertbutoxycarbonylhexyl-2,3,4,6-tetra-O-benzyl-β-D-galactopyranoside

To a stirred solution of 3.54 g (5.0 mmol) of 1-tert-butoxycarbonylhexane 6-ol in 25 ml of absolute 1,2-dichloroethane is added at room temperature each other finely powdered molecular sieve 4 Å (5.0 g) and silver silicate (5.0 g). On finally, dropwise at 0 ° C with a solution of 3.02 g (5.0 mmol) 2,3,4,6-tetra-O-benzyl-α-D-galactopyranosylbromide, dissolved in 40 ml absolute 1,2-dichloroethane. The reaction solution thus obtained becomes then stirred for 3 hours at -10 ° C. The mixture is worked up at 0 ° C mixed with 250 ml of 1,2-dichloroethane and the reaction solution on Celite sucked. The filtrate is washed in succession with 200 ml of aqueous sodium hydro gene carbonate solution and 220 ml of water, washed over sodium sulfate dries and filtered. After removing the solvent in vacuo the remaining residue is chromatographed on silica gel (eluent: toluene: Ethyl acetate = 10: 1).

449.6 mg (12.7% of theory) of the above-mentioned title compound are obtained, as a colorless oil.

Elemental analysis:
calc .: C 76.27; H 7.96;
Found: C 76.41; H 7.80;

b) 6-carboxyhexyl-2,3,4,6-tetra-O-benzyl-β-D-galactopyranoside

To 32.4 g (45.81 mmol) of the title compound from Example 52a) dissolved in 300 ml of 1,2-dichloroethane, 50 ml of trifluoroacetic acid are added at -10 ° C. and the mixture is stirred for 2 hours at this temperature. Then stirred for one hour at 0 ° C. It is evaporated to the dry state in a vacuum and the residue is chromatographed on silica gel (mobile solvent: n-hexane / 2-propanol = 5: 1).
Yield: 3.52 g (11.8% of theory) of a glassy solid.

Elemental analysis:
calc .: C 75.43; H 7.41;
Found: C 75.56; H 7.32;

c) Amide conjugate of the 24-polyamine based on the N, N, N ', N', N ", N" - Hexakis [2- (trilysyl-amino) -ethyl] trimesic acid triamide with the Gadolinium complex of 10- (4-carboxy-1-methyl-2-oxo-3-azabutyl) -1,4,7, 10-tetraazacyclododecane-1,4,7-triacetic acid and 6-carboxyhexyl-2,3,4,6- tetra-O-benzyl-β-D-galactopyranoside (m = 10, n = 14)

9.45 g (15 mmol) of the Gd complex of 10- (4-carboxy-1-methyl-2-oxo-3-azabutyl) -1,4,7,10-tetraazacyclododecane-1 described in Example 31h), 4,7-triacetic acid, 1.54 g of sodium bromide (15 mmol) and 8.28 g (72 mmol) of N-hydroxy succinimide and 14.88 g (21 mmol) of 6-carboxyhexyl-2,3,4,6-tetra- O-benzyl-β-D-galactopyranoside are dissolved in 200 ml of dimethyl sulfoxide at 50 ° C. After cooling to room temperature, 14.86 g (72 mmol) of N, N'-dicyclohexyl carbodiimide are added and the mixture is preactivated for 60 minutes. A solution of 1.386 g (0.5 mmol) of the 24-mer polyamine based on the N, N, N ', N', N ", N" -Hexakis is added to the N-hydroxysuccinimide ester solution thus prepared [2 - (Trilysyl-amino) -ethyl] -trimesic acid triamide and 7.28 g (72 mmol) of triethylamine in 10 ml of water and stirred overnight at room temperature. The suspension obtained is filtered and then sufficient acetone is added to complete the precipitation, the precipitate is filtered off with suction, dried, taken up in water, filtered off from the insoluble dicyclohexylurea and the filtrate is filtered through an AMICON® YM-3 ultrafiltration membrane (cut off 3000 Da) desalted and cleaned of low molecular components. The retentate is then freeze-dried.
Yield: 8.55 g (96% of theory)
H ₂ O content (Karl Fischer): 9.2%

Elemental analysis (calculated on anhydrous substance):
calc .: C 59.30; H 6.52; N 7.47; Gd 8.83;
Found: C 59.15; H 6.69; N 7.38; Gd 8.74;

A ratio of m: n = 10: 14 can be calculated from this.

d) amide conjugate of the 24-polyamine based on the N, N, N ', N', N ", N" - Hexakis [2- (trilysyl-amino) -ethyl] trimesic acid triamide with the Gadolinium complex of 10- (4-carboxy-1-methyl-2-oxo-3-azabutyl) -1,4,7, 10-tetraazacyclododecane-1,4,7-triacetic acid and 6-carboxyhexyl-β-D- galactopyranoside (m = 10, n = 14)

8.0 g (0.449 mmol) of the title compound from Example 52c) are dissolved in a mixture of 75 ml. Ethanol / 100 ml water and 2 g palladium catalyst (10% Pd on activated carbon) are added. It is hydrogenated for 12 hours at room temperature and a hydrogen pressure of 1 atm. The catalyst is filtered off and the filtrate is evaporated to dryness in vacuo. The residue is dissolved in 300 ml of water and freeze-dried.
Yield: 5.67 g (99% of theory) of a glassy solid.
H ₂ O content (Karl Fischer): 10.5%

Elemental analysis (calculated on anhydrous substance):
calc .: C 45.85; H 6.44; N 10.43; Gd 12.33;
Found: C 45.74; H 6.55; N 10.39; Gd 12.23;

A ratio of m: n = 10: 14 can be calculated from this.

Example 53 Amide conjugate of 24-polyamine based on the N, N, N ', N', N ", N" - Hexakis [2- (trilysyl-amino) -ethyl] trimesic acid triamide with the Gadolinium complex of 10- (4-carboxy-1-methyl-2-oxo-3-azabutyl) -1,4,7, 10-tetraazacyclododecane-1,4,7-triacetic acid and 6-carboxyhexyl-α-D- mannopyranoside (m = 7, n = 17) a) 6-Tertbutoxycarbonyl-hex-1-ol-2,3,4,6-tetra-O-benzyl-α-D-mannopyranoside

In an analogous manner to that described for the title compound from Example 52a), performs the reaction of 3.54 g (5.0 mmol) of 1-tert-butoxycarbonylhexan-6-ol with 3.02 g (5.0 mmol) 2,3,4,6-tetra-O-benzyl-α-D-mannopyranosyl bromide in absolute 1,2-dichloroethane at room temperature to form 368.5 mg (11.9% of theory) of the title compound as a colorless oil.

Elemental analysis:
calc .: C 76.27; H 7.96;
Found: C 76.38; H 7.89;

b) 1-carboxymethyl-2,3,4,6-tetra-O-benzyl-α-D-mannopyranoside

50 ml of trifluoroacetic acid are added to 30 g (45.81 mmol) of the title compound from Example 53a) dissolved in 300 ml of 1,2-dichloroethane at -10 ° C. and the mixture is stirred for 2 hours at this temperature. The mixture is then stirred at 0 ° C for one hour. It is evaporated to the dry state in a vacuum and the residue is chromatographed on silica gel (mobile solvent: n-hexane / 2-propanol = 5: 1).
Yield: 3.86 g (14.6% of theory) of a glassy solid.

Elemental analysis:
calc .: C 75.43; H 7.41;
found C 75.40; H 7.54;

c) Amide conjugate of the 24-polyamine based on the N, N, N ', N', N ", N" - Hexakis [2- (trilysyl-amino) -ethyl] trimesic acid triamide with the Gadolinium complex of 10- (4-carboxy-1-methyl-2-oxo-3-azabutyl) -1,4,7, 10-tetraazacyclododecane-1,4,7-triacetic acid and 6-carboxyhexyl-2,3,4,6- tetra-O-benzyl-α-D-mannopyranoside (m = 7, n = 17)

6.61 g (10.5 mmol) of the Gd complex of 10- (4-carboxy-1-methyl-2-oxo-3-azabutyl) -1,4,7,10-tetraazacyclododecane described in Example 31 h) -1,4,7- triacetic acid, 1.08 g sodium bromide (10.5 mmol), 8.28 g (72 mmol) N-hydroxy succinimide and 16.70 g (25.5 mmol) the title compound from Example 53b) are dissolved in 200 ml of dimethyl sulfoxide at 50 ° C. After cooling to room temperature, 14.86 g (72 mmol) of N, N'-dicyclohexylcarbodiimide are added and the mixture is preactivated for 60 minutes. A solution of 1.386 g (0.5 mmol) of the 24-mer polyamine based on the N, N, N ', N', N ", N" -Hexakis [2] is added to the N-hydroxy succinimide ester solution thus prepared - (trilysyl-amino) - ethyl] -trimesic acid triamide and 7.28 g (72 mmol) of triethylamine in 10 ml of water and stirred overnight at room temperature. The suspension obtained in this way is filtered and then sufficient acetone is added until precipitation is complete, the precipitate is filtered off with suction, dried, taken up in water, filtered off from the insoluble dicyclohexylurea and the filtrate is desalted via an AMICON® YM-3 ultrafiltration membrane (cut off 3000 Da) and cleaned of never dermolecular components. The retentate is then freeze-dried.
Yield: 8.49 g (95% of theory)
H ₂ O content (Karl Fischer): 7.6%

Elemental analysis (calculated on anhydrous substance):
calc .: C 63.28; H 6.76; N 6.27; Gd 6.16;
Found: C 63.17; H 6.83; N 6.15; Gd 6.09;

A ratio of m: n = 7:17 can be calculated from this.

d) amide conjugate of the 24-polyamine based on the N, N, N ', N', N ", N" - Hexakis [2- (trilysyl-amino) -ethyl] trimesic acid triamide with the Gadolinium complex of 10- (4-carboxy-1-methyl-2-oxo-3-azabutyl) -1,4,7,10- tetraazacyclododecane-1,4,7-triacetic acid and 6-carboxyhexyl-α-D- mannopyranoside (m = 7, n = 17)

8.0 g (0.447 mmol) of the title compound from Example 53c) are dissolved in a mixture of 75 ml of ethanol / 100 ml of water and 2 g of palladium catalyst (10% Pd on activated carbon) are added. It is hydrogenated for 12 hours at room temperature and a hydrogen pressure of 1 atm. The catalyst is filtered off and the filtrate is evaporated to dryness in vacuo. The residue is dissolved in 300 ml of water and freeze-dried.
Yield: 5.21 g (99% of theory) of a glassy solid.
H ₂ O content (Karl Fischer): 12%

Elemental analysis (calculated on anhydrous substance):
calc .: C 47.63; H 6.78; N 9.54; Gd 9.37;
Found: C 47.52; H 6.88; N 9.48; Gd 9.28;

A ratio of m: n = 7:17 can be calculated from this.

Example 54 24-cascade amine (Gd-Gly-Me-DOTA amide) ₁₁ - (N-glutaroyl-2-aminoethyl-1-thio-β-D-lactose amide) ₁₃ conjugate a) N-Glutaroyl-2-aminoethyl-1-thio-β-D-lactose-2,3,6,2 ", 3", 4 ", 6" -heptaacetate

To a stirred solution of 12.0 g (15 mmol) (2-aminoethyl) -1-thio-β-D- lactose-heptaacetate (for synthesis see: Yoshiaki Miura, Tsutomu Arai, Tatsuya Yamagata, Carbohydrate Reserarch 289 (1996) 193-199) in 200 ml absolute Tetrahydrofuran is added at a total of 2.05 g (18 mmol) glutaric acid at 22 ° C anhydride added in portions and leaves the reaction solution after 12 hours Stir in room temperature.

For the preparation, the solvent is removed in vacuo and the ver remaining residue with dichloromethane (250 ml) and 1 N hydrochloric acid (100 ml) transferred. After separating the organic product phase, it is still washed twice with 75 ml of water and dried over sodium sulfate net. The desiccant is filtered off and the solvent is removed in vacuo drawn. Crystallization from diethyl ether / pentane (1:10) yields 13.86 g (92.4% d. Th.) Of the title compound as a colorless and crystalline solid.

Elemental analysis:
calc .: C 49.01; H 5.73; N 1.73; S 3.96;
Found: C 49.10; H 5.89; N 1.83; S 3.92;

b) 24-cascade amine (Gly-Me-DOTA-tri-t-butyl ester amide) ₁₁ - [N-glutaroyl- (2-aminoethyl-1-thio-β-D-lactose-2,3,6, 2 ", 3", 4 ", 6" -hepta acetyl) amide] ₁₃ conjugate

14 mg (18.96 mmol) of the compound from Example 54a) are at room temperature rature in 100 ml of absolute dimethylformamide and at 0 ° C with 2.56 g  (22.2 mmol) N-hydroxysuccinimide followed by 4.55 g (22.2 mmol) dicyclo hexylcarbodimide added. After a reaction time of 60 minutes at 0 ° C and 3 hours at 22 ° C., the insoluble dicyclohexylurea is filtered off and drops the clear active ester solution of the above title ver bond slowly at 0 ° C to a stirred solution of 4.0 g (0.446 mmol; corresponding to 5.8 mmol of free amine functions) of the title compound from Bei game 10a). After a reaction time of 12 hours at room temperature the solvent removed in vacuo and with dichloromethane on a Ge filled up to a total volume of 500 ml. The organic phase is successively twice with water and twice with saturated sodium hydrogen carbonate washed solution and dried over sodium sulfate. After filtering the solvent is removed to dryness in vacuo. 7.7 g are obtained (92.6% of theory) of the title compound as a colorless oil. The ninhydrin reaction as TNBS determination of the title compound is also negative, i. H. in the bum lymer has no free amino functions. From the percentage sw The elemental analysis of the title compound gives a degree of loading of the polymer with 53.4% lactose heptaacetate units (corresponding to 13 Lactose esters per molecule).

Elemental analysis:
calc .: C 57.35; H 7.74; N 8.41;
Found: C 57.41; H 7.78; N 8.37;

c) 24-cascade amine (Gly-Me-DOTA amide) ₁₁ - (N-glutaroyl-2-aminoethyl-1-thio-β-D-lactose amide) ₁₃ conjugate

5.87 g (0.312 mmol) of the title compound from Example 54b) are dissolved in 100 ml absolute dichloromethane and then at 0 ° C with a total of 75 ml Trifluoroacetic acid added dropwise. After a response time of 12 hours is evaporated to dryness at room temperature in vacuo. The ver remaining residue is mixed with 100 ml of water and again in vacuo peeled off to dryness. The residue is dissolved in 100 ml of methanol and at room temperature with 50 ml of a 32% aqueous ammonia solution puts. After a reaction time of 12 hours at room temperature Vacuum removed from the solvent and the remaining residue in 200 ml of distilled water dissolved. The aqueous product solution is twice with each  Weil extracted 60 ml of diethyl ether and then by adding 10 % hydrochloric acid brought to pH 3.0. It is filled with water to a total volume of 800 ml and three times using a YM3 ultrafiltration membrane bran (AMICON®) ultrafiltered against distilled water. The remaining one Residue is made up to a volume of 500 ml with deionized water filled and freeze-dried.

Elemental analysis:
calc .: C 47.58; H 6.61; N 8.27; S 3.62;
Found: C 47.61; H 6.58; N 8.32; S 3.68;
Water content: 6.43%
Yield: 3.29 (89.6% of theory) as an amorphous and colorless powder.

An average degree of loading of the Polymers of 45.4% Gly-Me-DOTA and 54.6% lactose (corresponding to 11 Gly- Me-DOTA units and 13-D-lactose residues per molecule). In the Ti are not free amino functions by means of the ninhydrin reaction more detectable.

d) 24-cascade amine (Gd-Gly-Me-DOTA amide) ₁₁ - (N-glutaroy-2-aminoethyl-1-thio-β-D-lactose amide) ₁₃ conjugate

2.53 g (0.22 mmol; corresponding to 2.42 mmol of Gly-Me-DOTA) of the title compound solution from Example 54c) are dissolved in 70 ml of distilled water and the pH The value of the solution is determined by dropwise addition with 10% aqueous salt acidity set to 5.0. Finally, 670 mg are added at room temperature (2.55 mmol) gadolinium chloride and stir the reaction solution for 12 Hours at 60 ° C. At room temperature, the reaction solution by Ver put with 1 molar sodium hydroxide solution to pH 7.2 and with distilled Water made up to a total volume of 500 ml. After three times ultra filtration against distilled water over a YM3 ultrafiltration membrane (AMICON®) the remaining residue is freeze-dried. You get 2.7 g (98.6% of theory) of the title compound as an amorphous, colorless powder with a Water content of 6.12%.  

Elemental analysis (calculated on anhydrous substance):
calc .: C 41.47; H 5.51; N 7.21; S 3.16; Gd 13.10;
Found: C 41.54; H 5.60; N 7.16; S 3.09; Gd 13.18;

The molar element ratio of sulfur to gadolinium results thus an average degree of loading of the polymer of 45.2% Gd-Gly-Me-DOTA and 54.8% lactose (corresponding to 11 Gd-Gly-Me-DOTA units and 13-D- Lactose residues per molecule). In the title link are using ninhydrin crack tion no free amino functions detectable.

Example 55 24-cascade amine (Gd-Gly-Me-DOTA amide) ₁₁ - (N-glutaroyl-2-aminoethyl-1-O-β-D-lactose amide) ₁₃ conjugate a) N-Glutaroyl-2-aminoethyl-1-O-β-D-lactose-2,3,6,2 ", 3", 4 ", 6" -heptaacetate

In an analogous manner as described for Example 54a), the implementation of 14.0 g (21.6 mmol) (2-aminoethyl) -β-latosideheptaacetat (Yoshiaki Miura, Tsu tomu Arai, Tatsuya Yamagata, Carbohydrate Reserarch 289 (1996) 193-199) in 250 ml of anhydrous THF with 2.96 g (26 mmol) of glutaric anhydride Working up 14.9 g (87.0% of theory) of the title compound as a colorless solid.

Elemental analysis (calculated on anhydrous substance):
calc .: C 50.00; H 5.85; N 1.77;
Found: C 49.89; H 5.92; N 1.80;

b) 24-cascade amine (Gly-Me-DOTA-tri-t-butyl ester amide) ₁₁ - [N-glutamoyl- (2-aminoethyl-1-O-β-D-lactose-2,3,6 , 2 ", 3", 4 ", 6" -heptaacetyl) amide] ₁₃ conjugate

14.4 g (18.16 mmol) of the compound from Example 55a) are at room temperature rature in 100 ml of absolute dimethylformamide and at 0 ° C with 2.51 g (21.8 mmol) N-hydroxysuccinimide followed by 4.46 (28.8 mmol) dihydro  hexylcarbodimide added. After a reaction time of 60 minutes at 0 ° C and 3 hours at 22 ° C., the insoluble dicyclohexylurea is filtered off and drops the clear active ester solution of the above title ver bond slowly at 0 ° C to a stirred solution of 3.9 g (0.435 mmol; corresponding to 5.65 mmol of free amine functions) from the title compound Example 10a). After a reaction time of 12 hours at room temperature the solvent is removed in vacuo and with dichloromethane Total volume of 500 ml filled up. The organic phase is successively the twice with water and twice with saturated sodium bicarbonate washed solution and dried over sodium sulfate. After filtering the solvent is removed to dryness in vacuo. 7.48 g are obtained (90.4% of theory) of the title compound as a colorless oil. The ninhydrin reaction as TNBS determination of the title compound is also negative, i. H. in the bum lymer has no free amino functions. From the percentage sw The elemental analysis of the title compound gives a degree of loading of the polymer with 54.5% lactose heptaacetate units (corresponding to 13 Lactose residues per molecule.)

Elemental analysis:
calc .: C 56.72; H 7.65; N 8.31;
Found: C 56.82; H 7.69; N 8.41;

c) 24-cascade amine (Gly-Me-DOTA amide) ₁₁ - (N-glutaroyl-2-aminoethyl-1-O-β-D-lactose amide) ₁₃ conjugate

5.93 g (0.312 mmol) of the title compound from Example 55b) are dissolved in 100 ml of pure dichloromethane and a total of 75 ml of trifluoroacetic acid are 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 oily residue thus obtained 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 in each case 60 ml of diethyl ether and then brought to pH 3.0 with 10% hydrochloric acid. It is 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.
Water content: 9.30%
Yield: 3.23 g (91.7% of theory) as an amorphous and colorless powder.

Elemental analysis (calculated on anhydrous substance):
calc .: C 48.46; H 6.73; N 7.33;
Found: C 48.52; H 6.59; N 7.40;

d) 24-cascade amine (Gd-Gly-Me-DOTA amide) ₁₁ - (N-glutaroyl-2-aminoethyl-1-O-β-D-lactose amide) ₁₃ conjugate

2.48 g (0.22 mmol; corresponding to 2.42 mmol of Gly-Me-DOTA) of the title compound solution from Example 55c) are dissolved in 70 ml of distilled water and the pH The value of the solution is determined by dropwise addition with 10% aqueous salt acidity set to 5.0. Then 670 mg are added at room temperature (2.55 mmol) gadolinium chloride, dissolved in 10 ml of distilled water and stirred the reaction solution at 60 ° C for 12 hours. At room temperature the reaction solution is adjusted to pH 7.2 by adding 1 molar sodium hydroxide solution brought and with distilled water to a total volume of 500 ml filled. After ultrafiltration three times against distilled water over a YM3 ultrafiltration membrane (AMICON®) becomes the remaining residue freeze-dried. 2.75 g (96.1% of theory) of the title compound are obtained as amorphous, colorless powder with a water content of 4.19%.

Elemental analysis (calculated on anhydrous substance):
calc .: C 42.13; H 5.50; N 7.33; Gd 13.31;
Found: C 42.20; H 5.60; N 7.26; Gd 13.28;

The molar element ratio of sulfur to gadolinium results thus an average degree of loading of the polymer of 45.7% Gd-Gly-Me-DOTA and 54.3% lactose (corresponding to 11 Gd-Gly-Me-DOTA units and 13-D- Lactose residues per molecule). In the title compound are quantitative Ninhydrin reaction no longer detectable free amino functions.  

Example 56 24-cascade amine (Gd-DTPA amide) ₁₁ - (α-D-mannopyranose-1-thio-3-propionate amide) ₁₃ conjugate a) 24-cascade amine (Gd-DTPA-tetra-t-butyl ester amide) ₁₁

20.0 g (32.37 mmol) 3,9-bis (t-butoxycarbonylmethyl) -3,6,9-triazaundecandi carboxylic acid di-t-butyl ester (representation as in European patent applications solutions: EP 0430863; EP 0331616 and EP 0271180) are described in Room temperature dissolved in 100 ml of absolute dimethylformamide and with stirring 4.47 g (38.84 mmol) of N-hydroxysuccinimide were added. Then will the clear reaction mixture cooled to 0 ° C and porti at this temperature on a total of 7.97 g (38.84 mmol) of dicyclohexylcarbodiimide. After a reaction time of 30 minutes at 0 ° C and 2.5 hours in the room temperature is filtered off from the precipitated dicyclohexylurea. So he held filtrate is now slowly added dropwise at room temperature to a stirred one Solution of 9.0 g (1.9 mmol; corresponding to 44.8 mmol of free amino function nen) 24-cascade polyamine and 4.6 g (45.8 mmol) triethylamine in 60 ml abso lutem DMF. After a reaction time of 12 hours at room temperature evaporated to dryness in vacuo and the remaining residue with 250 ml of dichloromethane are added. After filtering off insoluble matter the organic phase successively with 5% hydrochloric acid, saturated sodium hydrogen carbonate solution and washed twice with water. After this Drying the organic product phase over sodium sulfate is filtered and the Stripped off solvent in vacuo. 14.0 g (79.2% of theory; based on to the polyamine used) of a slightly yellow colored oil, which without further cleaning immediately for the subsequent synthesis steps is set. After quantitative ninhydrin reaction of the above title ver Binding there are an average of 13 free amine functions per molecule.

Elemental analysis:
calc .: C 58.84; H 9.01; N 11.66;
Found: C 57.79; H 9.24; N 11.80;

b) 24-cascade amine (DTPA-tetra-tert-butyl ester amide) ₁₁ - [(3- (2,3,4, 6-tetra-O-acetyl) -1-thio-α-D-mannopyranosyl-3 -propion-amide) ₁₃ - conjugate

A stirred solution of 3.5 g (0.27 mmol; corresponding to 3.5 mmol of free Amine functions) of the title compound from Example 56a) in 100 ml of absolute Dichloromethane is at room temperature with 617 mg (6.1 mmol) of triethylamine transferred. Then with a solution of 2.97 g (5.61 mmol) of 3- (2,3,4,6- Tetra-O-acetyl-1-thio-α-D-mannopyranosyl) propionic acid N-hydroxysuccinimide ester (Lee, Y.C. et al., Biochemi., Vol. 15, No. 18, 1976, 3956-3963; Krohn, K.A. et al., J. Nucl. Med., Vol. 26, 10, 1985, 1157-1167) in 60 ml dichloromethane added dropwise. After a reaction time of 12 hours at room temperature temperature is made up to a total volume of 500 ml with dichloromethane. The organic phase is successively twice with water and twice with saturated sodium bicarbonate solution and washed over sodium sulfate dried. After filtering, the solvent is removed in vacuo to Dry peeled. 3.55 g (92.6% of theory) of the title compound are obtained as colorless oil. The ninhydrin reaction of the title compound is negative, i. H. in the Free amino functions are no longer detectable in polymer.

Elemental analysis:
calc .: C 55.10; H 7.73; N 7.37; S 2.81;
Found: C 55.20; H 7.78; N 7.31; S 2.79;

From the percentage sulfur value of the elemental analysis of the title compound there is an average degree of loading of the polymer with 55% D-mannose tetraacetate (corresponding to 13 D-mannose residues per molecule).

c) 24-cascade amine (DTPA amide) ₁₁ - (α-D-mannopyranose-1-thio-3-propione amide) ₁₃ conjugate

4.5 g (0.30 mmol) of the title compound from Example 56b) 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 concentrated 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 at room temperature with 50 ml of a 32% aqueous ammonia solution. 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 and then brought to pH 3.0 by adding 10% hydrochloric acid, 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: 2.80 g (81.3% of theory) as an amorphous and colorless powder.
Water content: 6.39%

Elemental analysis (calculated on anhydrous substance):
calc .: C 43.20; H 5.65; N 9.82; Na 9.09; S 3.75;
Found: C 43.22; H 5.71; N 9.79; Na 9.12; S 3.70;

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

d) 24-cascade amine (Gd-DTPA amide) ₁₁ - (α-D-mannopyranose-1-thio-3-propionamide) ₁₃ conjugate

2.5 g (0.22 mmol; corresponding to 2.47 mmol of DTPA) of the title compound Example 56c) are dissolved in 70 ml of distilled water and the pH of the Solution is added by dropwise addition with 10% aqueous hydrochloric acid 5.2 set. Finally, a solution of is added at room temperature Add 670 mg (2.55 mmol) of gadolinium chloride in 10 ml of distilled water and stir the reaction solution for 12 hours at room temperature. The Reak tion solution is added to pH 7.2 by adding 1 molar sodium hydroxide solution brought and made up with distilled water to a total volume of 500 ml fills. After three times ultrafiltration against distilled water over a YM3  Ultrafiltration membrane (AMICON®) freezes the remaining residue dried. 2.6 g (98.3% of theory) of the title compound are obtained as an amorphous, colorless powder with a water content of 8.11%.

Elemental analysis (calculated on anhydrous substance):
calc .: C 39.73; H 5.20; N 9.03; S 3.45; Gd 14.30;
Found: C 39.81; H 5.20; N 9.07; S 3.49; Gd 14.28;

From the molar elementary ratios of sulfur to gadolinium thus an average degree of loading of the polymer of 45.2% Gd-DTPA and 54.8% D-mannose (corresponding to 11 Gd-DTPA units and 13-D-mannose residues per molecule). There are no ninhydrin reactions in the title compound free amino functions more detectable.

Example 57 24-cascade amine (Gd-DTPA amide) _11- (β-D-galactopyranose-1-thio-3-propionate amide) ₁₃ conjugate a) 24-cascade amine (DTPA-tetra-tert-butyl ester amide) ₁₁ - [(3- (2,3,4,6-tetra-O-acetyl) -1-thio-β-D-galactopyranosyl-3 -propion-amide) ₁₃ conjugate

In an analogous manner as described for Example 56b), the implementation of 3.0 g (0.32 mmol; corresponding to 4.2 mmol of free amine functions) of the title Compound from Example 56a) with 2.76 g (5.2 mmol) of 3- (2,3,4,6-tetra-O-acetyl-1- thio-β-D-galactopyranosyl) propionic acid N-hydroxysuccinimide ester (representation according to: Lee, Y.C. et al., Biochemi., Vol. 15, No. 18, 1976, 3956-3963; Krohn, K.A. et al., J. Nucl. Med., Vol. 26, 10, 1985, 1157-1167) to form 4.15 g (87.6% of theory) of the above-mentioned title compound as a colorless oil.

Elemental analysis:
calc .: C 55.10; H 7.73; N 7.37; S 2.81;
Found: C 55.14; H 7.80; N 7.31; S 2.88;

The title compound shows a negative ninhydrin reaction. From the percent len sulfur value of the elementary analysis results in an average degree of loading of the polymer with 13 D-galactose residues per molecule.

b) 24-cascade amine (DTPA amide) ₁₁ - (β-D-galactopyranose-1-thio-3-propione amide) ₁₃ conjugate

In an analogous manner as described for example 56c), the deprotection of 3.5 g (0.23 mmol) of the title compound from example 56b) after freeze-drying leads to the formation of 2.26 g (88.6% of theory). ) the above-mentioned title compound as an amorphous powder.
Water content: 5.74%

Elemental analysis (calculated on anhydrous substance):
calc .: C 43.20; H 5.65; N 9.82; Na 9.09; S 3.75;
Found: C 43.22; H 5.68; N 9.77; Na 9.12; S 3.70;

c) 24-cascade amine (Gd-DTPA amide) ₁₁ - (β-D-galactopyranose-1-thio-3-propione amide) ₁₃ conjugate

In an analogous manner to that described for Example 56d), the complexation of 2.0 g (0.180 mmol; corresponding to 1.97 mmol of DTPA) of the title compound from Example 57b) with 525 mg (2.00 mmol) of gadolinium chloride, dissolved in 10 ml of distilled water to form the above title compound.
Yield: 2.14 g (98.4% of theory) of the title compound as a colorless and amorphous powder
Water content: 7.22%

Elemental analysis (calculated on anhydrous substance):
calc .: C 39.73; H 5.20; N 9.03; Na 2.09; S 3.45; Gd 14.30;
Found: C 39.84; H 5.22; N 9.10; Na 2.12; S 3.40; Gd 14.27;

Average degree of loading per molecule of the above title compound:
11 Gd DTPA units
and 13 D-galactopyranosyl residues

Example 58 24-cascade polyamine ( ¹¹¹ indium DTPA amide) ₈ - [β- (1-deoxy-1-thioureido) sialyl-Le ^x ] ₁₆ conjugate a) 24-cascade polyamine (DTPA-tetra-t.butyl ester amide) ₈ conjugate

12.35 g (20.00 mmol) 6,9-bis (t-butoxycarbonylmethyl) -3-carboxymethyl-3,6,9- triazaundecanedicarboxylic acid di-t-butyl ester at room temperature in 100 ml of absolute dimethylformamide dissolved and with stirring with 2.9 g (24 mmol) of N- Hydroxysuccinimide added. Then the clear reaction mixture Cooled 0 ° C and at this temperature in portions with a total of 49 g (24 mmol) dicyclohexylcarbodiimide added. After a response time of 30 Minutes at 0 ° C and 2.5 hours at room temperature is from the failed Filtered off dicyclohexylurea. The filtrate obtained in this way is now slowly added dropwise at room temperature to a stirred solution of 9.0 g (1.9 mmol; chend 45.8 mmol of free amino functions) 24-cascade polyamine and 4.6 g (45.8 mmol) triethylamine in 60 ml absolute DMF. After a response time from 12 hours at room temperature is vacuumed to dryness concentrated and the remaining residue with 250 ml of dichloromethane. After after filtering off insoluble constituents, the organic phase becomes in succession the with 5% hydrochloric acid, saturated sodium bicarbonate solution twice with Washed water. After drying the organic product phase over Sodium sulfate is filtered and the solvent is removed in vacuo. Man easily obtains 12.38 g (94.6% of theory; based on salted polyamine) yellow colored oil, which without further cleaning immediately for the after following synthesis steps is used. After quantitative ninhydrin freak tion of the above-mentioned title compound have an average of 16 free amine functions per molecule.

b) 24-cascade polyamine (DTPA amide) ₈ conjugate

4.5 g (0.58 mmol) of the title compound from Example 58a) 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 concentrated 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 200 ml of distilled water. The aqueous product solution is extracted twice with 60 ml of diethyl ether each. The aqueous product solution is then brought to pH 3.0 by adding 10% hydrochloric acid, made up to a total volume of 800 ml with water and ultrafiltered three times using a YMA ultrafiltration membrane (AMICON®) against distilled water. 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. After renewed ultrafiltration, the remaining residue is made up to a volume of 500 ml with deionized water and freeze-dried.
Yield: 3.16 g (84.3% of theory) as an amorphous and colorless powder.
Water content: 7.33%

Elemental analysis (calculated on anhydrous substance):
calc .: C 44.70; H 6.0; N 14.92; Na 11.36;
Found: C 44.77; H 6.6; N 14.90; Na 11.31;

c) 1-deoxy-1-isothiocyanato-sialyl-Le ^x -

270 mg (0.293 mmol) 1-deoxy-1-amino-sialyl-Le ^x (representation according to: Kunz, H .; et al., Angew. Chem. 1994, 106. No. 20; Kunz, H .; et al ., Angew. Chem. 1996, 108, No. 3, 359-362; Kunz, H. et al., Angew. Chem. 1995, 107, No. 9; Kretzschmar et al., Tetrahedron, Vol. 51, No . 47, 13015-13030, 1995) are dissolved in 40 ml of 80% aqueous ethanol and 0.056 ml (0.73 mmol) of thiophosgene are added at 0 ° C. by addition via a microsyringe. After a reaction time of 1 hour at 22 ° C, the ethanol is removed in vacuo and the aqueous residue of the crude isothiocyanate obtained is used directly for the subsequent reaction, the content of the above-mentioned title compounding the aqueous crude product solution thus obtained with 0.276 mmol (corresponds to 95% conversion) is assumed, which are contained in a total volume of 7.6 ml of aqueous solution.

Thin layer chromatography (ethyl acetate / methanol / water / glacial acetic acid = 5: 3: 3: 0.5; R _f = 0.54) of the reaction solution indicates complete and uniform conversion to the above-mentioned title compound.

Sulfur determination (based on anhydrous sample):
calc .: S 3.31;
found: S 3.24;

d) 24-cascade polyamine (DTPA amide) ₈ - [β- (1-deoxy-1-thioureido) sialyl-Le ^x ] ₁₆ conjugate

The reaction of 54.5 mg (0.00842 mmol; corresponding to 0.134 mmol of free amino functions) of the title compound from Example 58b) with the entire aqueous solution of the crude product of the title compound from Example 58c, corresponding to 0.276 mmol), yields 164.7 after working up and freeze-drying mg (89% of theory) of an amorphous and colorless solid.
Water content: 10.01%

Elemental analysis (determination of sodium and sulfur; based on water-free sample):
calc .: S 2.34; Na 3.35;
found: S 2.36; Na 3.29;

The ninhydrin reaction as well as the TNBS determination of the title compound are negative, ie there are no free amino functions in the polymer. The percentage sulfur value of the elemental analysis of the title compound gives a degree of loading of the polymer with 67.4% sialyl-Le ^x (corresponding to 8 sialyl-Le ^x residues per molecule). The degree of loading of the above-mentioned title compound with 67.2% of tetrasaccharide (corresponding to 16 sialyl-Le ^x residues per molecule) is determined from the quantitative determination of sulfuric acid using TBA (thiobarbituric acid) by a procedure known from the literature (cf. Example 39). After another colorimetric titration of the title compound 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 of 32.8% (corresponding to 8 DTPA units per molecule) could be confirmed.

e) 24-cascade polyamine ( ¹¹¹ indium DTPA amide) ₈ - [β- (1-deoxy-1-thioureido) sialyl-Le ^x ] ₁₆ conjugate

1 mg of the title compound from Example 58d) is dissolved in 2 ml of sodium citrate buffer (pH 5.3) and a solution of ¹¹¹ indium chloride (initial activity: 17.2 MBq) is added at room temperature. 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 and sterile filtered.

The solution thus produced is direct for the corresponding radiodiagnosti purposes

Example 59 24-cascade polyamine ( ¹⁵⁷ gadolinium DTPA amide) ₈ - [β-1-O- (6-thioureidohexyl) sialyl-Le ^x ] ₁₆ conjugate a) 24-cascade polyamine (DTPA amide) ₈ conjugate

5.0 g (1.80 mmol; corresponding to 43.2 mmol of free amino functions) 24-cascade polyamine are dissolved in 400 ml of distilled water and adjusted to a pH of 9.5 with 1 molar sodium hydroxide solution. A total of 8.0 g (20.0 mmol) of DTPA monoanhydride monoethyl ester is then added, 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 resulting aqueous product solution 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: 10.63 g (91.2% of theory; based on the polyamine used) as an amorphous powder
Water content: 7.88%

Elemental analysis (calculated on anhydrous substance):
calc .: C 44.70; H 6.04; N 14.92; Na 11.36;
Found: C 44.81; H 6.10; N 14.90; Na 11.32;

After elemental analysis as well as colorimetric titration with 0.0025 molar Gadolinium sulfate standard solution in the presence of xylenol orange as an indicator, this results in a DTPA loading level of the polymer of 33.47% (corresponding 8 DTPA units per molecule). After quantitative ninhydrin reaction as well the quantitative determination of amino groups with the TNBS method are in the statistical means per polymer molecule 16 free amino functions.

b) β-1-O- (6-isothiocyanatohexyl) sialyl-Le ^x

250 mg (0.243 mmol) β-1-O- [6-aminohexyl] sialyl-Le ^x (representation according to: Kunz, H .; et al., Angew. Chem. 1994, 106. No. 20; Kretzschmar et al ., Tetra hedron, Vol. 51, No. 47, 13015-13030, 1995) are dissolved in 30 ml of 80% aqueous ethanol and at 0 ° C. with 0.046 ml (0.60 mmol) of thiophosgene by addition via a microsyringe transferred. After a reaction time of 1 hour at 22 ° C, the ethanol is removed in vacuo and the aqueous residue of the crude isothiocyanate obtained is used directly for the subsequent reaction, the content of the title compound mentioned above corresponding to the aqueous crude product solution thus obtained at 0.23 mmol ( a 95% conversion) is assumed, which are contained in a total volume of 5.6 ml of aqueous solution.

Thin layer chromatography (ethyl acetate / methanol / water / glacial acetic acid = 5: 3: 3: 0.5; R _f = 0.72) of the reaction solution shows complete and uniform conversion to the above-mentioned title compound.

Elemental analysis (sulfur determination; based on an anhydrous sample):
calculated: S 3.00;
found: S 2.90;

c) 24-cascade polyamine (DTPA amide) ₈ - [β-1-O- (6-thioureidohexyl) sialyl-Le ^x ] ₁₆ conjugate

The reaction of 50.0 mg (0.00770 mmol; corresponding to 0.123 mmol of free amino functions) of the title compound from Example 59a) with the entire aqueous solution of the crude product of the title compound from Example 59b) (0.23 mmol) provides after working up and freeze drying 167 7 mg (92.4% of theory) of an amorphous and colorless solid.
Water content: 9.04%

Elemental analysis (determination of sodium and sulfur; based on water-free sample):
calc .: S 2.18; Na 6.95;
found: S 2.29; Na 7.03;

The ninhydrin reaction as well as the TNBS determination of the title compound are negative, ie there are no free amino functions in the polymer. The percentage sulfur value of the elemental analysis of the title compound gives a degree of loading of the polymer with 67.0% sialyl-Le ^x units (corresponding to 16 sialyl-Le ^x residues per molecule).

From quantitative sialic acid determination using TBA (thiobarbituric acid) using a procedure known from the literature [cf. Lit .: Aminoff, D., (1961), "Methods for the quantitative estimation of N-acetylneuraminic acid and their application to hydrolysates of sialomucoids", J. Biochem., 81, 384-392; Uschi da, Y. and Sugimori, T. (1977), "Distribution of neuraminidase in Arthrobacter and its purification by affinity chromatography", J. Biochem. (Tokyo), 82, 1425-1433] the degree of loading of the above-mentioned title compound with 68.1 of tetrasaccharide (corresponding to 16 sialyl-Le ^x residues per molecule) is determined. After another colorimetric titration of the title compound with 0.0025 molar standard gadolinium sulfate solution, in In the presence of xylenol orange as an indicator, the DTPA loading level of the polymer of 32.1% (corresponding to 8 DTPA units per molecule) was confirmed.

d) 24-cascade polyamine ( ¹⁵⁷ gadolinium DTPA amide) ₈ - [sialyl-Le ^x -1- (6-hexyl-thiourea)] ₁₆ conjugate

1 mg of the title compound from Example 59c) are dissolved in 2 ml of sodium citrate buffer (pH 5.3) and a solution of ¹⁵⁷ gadolinium chloride (initial activity: 16.2 MBq) is added at room temperature. 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 and sterile filtered. The solution prepared in this way can be used directly for the relevant radio diagnostic purposes

Example 60 24-cascade polyamine (Gd-DTPA amide) ₁₁ - (N-glutaroyl-3-aminoethyl-β-1-thiolactoside amide) ₁₃ conjugate a) 24-cascade polyamine (DTPA tetra-t-butyl ester amide) ₁₁ - (N-glutaroyl-3-aminoethyl-β-1-thiolactoside amide) ₁₃ conjugate

In an analogous manner to that described in Example 54b), the conversion of 5.40 g (0.58 mmol, 7.54 mmol of free amine) of the title compound from Example 56a), which was previously dissolved in 25 ml of absolute N, N-dimethylformamide and the solution so contained was cooled to 0 ° C, with a solution of 12.16 g (13.92 mmol) N-glutaroyl-3-aminoethyl-β-1-thiolactosylheptaacetate-glutaric acid - N-hydroxysuccinimide ester [(for illustration see Example 54b)] in 75 ml abso lutem N, N-dimethylformamide to form the desired title compound. This is done after a reaction time of 12 hours at room temperature Stripped solvent in vacuo and with dichloromethane to a total 500 ml lumen filled. The organic phase is processed successively twice with water and twice with saturated sodium hydro Gene carbonate solution washed and dried over sodium sulfate. After this The solvent is filtered off in vacuo to dryness. Man receives 10.0 g (90.1% of theory) of the title compound as a colorless oil. The ninhydrin reaction as well as the TNBS determination of the title compound are negative, d. H. there are no longer any free amino functions in the polymer. From the pro the central sulfur value of the elementary analysis of the title compound results  a degree of loading of the polymer with 52.9% lactose heptaacetate units (corresponding to 13 lactose residues per molecule).

b) 24-cascade polyamine (DTPA amide) ₁₁ - (N-glutaroyl-3-aminoethyl-β-1-thiolactoside amide) ₁₃ conjugate

6.1 g (0.312 mmol) of the title compound from Example 60a) are dissolved in 100 ml of pure dichloromethane and then a total of 75 ml of trifluoroacetic acid are added dropwise at 0 ° C. After a reaction time of 12 hours at room temperature, the mixture is concentrated to dryness in vacuo. The remaining residue is mixed with 100 ml of water and stripped again to dryness in vacuo. The oily residue thus obtained 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 and then brought to pH 3.0 by adding 10% hydrochloric acid. It is 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: 3.95 g (88.7% of theory) as an amorphous and colorless powder
Water content: 4.88%

Elemental analysis (calculated on anhydrous substance):
calc .: C 44.44; H 5.85; N 8.90; S 2.91; Na 7.06;
Found: C 44.49; H 5.92; N 8.79; S 2.87; Na 6.98;

c) 24-cascade polyamine (Gd-DTPA amide) ₁₁ - (N-glutaroyl-3-aminoethyl β-1-thiolactoside amide) ₁₃ conjugate

3.15 g (0.22 mmol; corresponding to 2.42 mmol of DTPA) of the title compound Example 60b) are dissolved in 70 ml of distilled water and the pH of the  Solution is added by dropwise addition with 10% aqueous hydrochloric acid 4.0 set. 670 mg (2.55 mmol) are then added at room temperature. Add gadolinium chloride, dissolved in 10 ml of distilled water, and stir Reaction solution for 12 hours at 60 ° C. At room temperature the reak tion solution brought to pH 7.2 by adding 1 molar sodium hydroxide solution made up to a total volume of 500 ml with distilled water. After three times ultrafiltration against distilled water via a YM3 ultrafiltra tion membrane (AMICON®), the remaining residue is freeze-dried net. 3.3 g (98.4% of theory) of the title compound are obtained as an amorphous, colorless Powder with a water content of 7.39%.

Elemental analysis (calculated on anhydrous substance):
calc .: C 41.71; H 5.28; N 8.35; Na 1.66; S 2.73; Gd 11.33;
Found: C 41.81; H 5.34; N 8.29; Na 1.62; S 2.69; Gd 11.40;

The molar element ratio of sulfur to gadolinium results an average degree of loading of the polymer of 46% DTPA and 54% lactose (corresponding to 11 Gd-DTPA units and 13 lactose residues per molecule). In of the title compound are no free amino functions by means of a ninhydrin reaction more detectable.

Example 61 48-cascade polyamine (Gd-DTPA amide) ₁₈ - (α-D-mannopyranosyl-1-thiopropionic acid amide) ₃₀ conjugate a) 48-cascade polyamine- [DTPA-tetra-t-butyl ester amide (terminal)] ₁₈ conjugate

16.55 g (26.8 mmol) of DTPA-tetra-tert-butyl ester (representation: German patent application no. 195 08 058 A) are absolute at room temperature in 100 ml dissolved dimethylformamide and with stirring with 3.7 g (32.16 mmol) of N-Hy droxysuccinimide added. Then the clear reaction mixture Cooled 0 ° C and at this temperature in portions with a total of 6.6 g (32.16 mmol) dicyclohexylcarbodiimide added. After a response time of 30  Minutes at 0 ° C and 2.5 hours at room temperature is from the failed Filtered off dicyclohexylurea. The filtrate obtained in this way is now slowly added dropwise at room temperature to a stirred solution of 6.5 g (0.99 mmol; correspond chend 47.8 mmol of free amino functions) 48-cascade polyamine and 4.8 g (50.0 mmol) triethylamine in 60 ml absolute DMF. After a response time from 12 hours at room temperature is vacuumed to dryness concentrated and the remaining residue with 250 ml of dichloromethane. After after filtering off insoluble constituents, the organic phase becomes in succession that with 5% hydrochloric acid, saturated sodium bicarbonate solution twice washed with water. After drying the organic product phase It is filtered over sodium sulfate and the solvent is removed in vacuo. 14.5 g (85.2% of theory; based on the polyamine used) of one are obtained light yellow colored oil, which can be used immediately without further cleaning subsequent synthesis steps is used.

After quantitative ninhydrin reaction of the above title compound gene on average 30 free amine functions per molecule.

Elemental analysis:
calc .: C 59.87; H 9.20; N 8.88;
Found: C 59.80; H 9.28; N 8.90;

b) 48 cascade polyamine- [DTPA-tetra-t-butyl ester amide (terminal)] ₁₈ - [3- (2,3,4,6-tetra-O-acetyl-α-D-mannopyranosyl) -1- thiopropionic acid amide] ₃₀ conjugate

A stirred solution of 3.26 g (0.19 mmol; corresponding to 5.7 mmol of free Amine functions) of the title compound from Example 61a) in 100 ml of absolute Dichloromethane is at room temperature with 657 mg (6.5 mmol) of triethylamine transferred. Then with a solution of 3.3 g (6.3 mmol) of 3- (2,3,4,6- Tetra-O-acetyl-1-thio-α-D-mannopyranosyl) propionic acid N-hydroxysuccinimide ester added dropwise in 60 ml of dichloromethane. After a response time of 12 hours at room temperature is made up to a total with dichloromethane volume of 500 ml filled. The organic phase becomes two in succession times with water and twice with saturated sodium bicarbonate solution washed and dried over sodium sulfate. After filtering it will Stripped solvent to dryness under vacuum. 5.07 g are obtained (90.3% of theory) of the title compound as a colorless oil. The ninhydrin reaction of  Title connection is negative, i. H. there are no free amino radio in the polymer tion more detectable.

Elemental analysis:
calc .: C 53.52; H 7.48; N 7.22; S 3.16;
Found: C 53.90; H 7.52; N 7.18; S 3.21;

From the percentage sulfur value of the elemental analysis of the title compound there is an average degree of loading of the polymer with 62.83% α-D-Manno se-tetraacetate units (corresponding to 30 α-D-mannose residues per molecule).

c) 48-cascade polyamine (DTPA amide) ₁₈ - (α-D-mannopyranosyl-1-thio-3-propionic acid amide) ₃₀ conjugate

5.17 g (0.17 mmol) of the title compound from Example 61b) are dissolved in 100 ml of solute 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 concentrated 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 at room temperature with 50 ml of a 32% aqueous ammonia solution. After a reaction time of 12 hours at room temperature, the solvent is removed in vacuo and the remaining residue is dissolved in 200 m) of distilled water. The aqueous product solution is extracted twice with 60 ml of diethyl ether each time. The aqueous product solution is brought to pH 3.0 by adding 10% hydrochloric acid and 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: 3.47 g (91.6% of theory) as an amorphous and colorless powder.
Water content: 7.34%

Elemental analysis (calculated on anhydrous substance):
calc .: C 43.91; H 6.09; N 9.82; S 4.29; Na 7.39;
Found: C 43.89; H 6.12; N 9.84; S 4.34; Na 7.42;

d) 48-cascade polyamine (Gd-DTPA amide) ₁₈ - (α-D-mannopyranosyl-1-thio-3-propionic acid amide) ₃₀ conjugate

3.04 g (0.136 mmol; corresponding to 2.45 mmol of DTPA of the title compound Example 61c) are dissolved in 70 ml of distilled water and the pH of the Solution by dropwise addition with 10% aqueous hydrochloric acid to 5.0 set. Then 655.8 mg (2.5 mmol) are added at room temperature. Gadolinium chloride, in the form of 10 ml of an aqueous solution, is added and stirred the reaction solution for 12 hours at 60 ° C. At room temperature the Reaction solution by adding 1 molar sodium hydroxide solution to pH 7.2 ge brought and made up with distilled water to a total volume of 500 ml fills. After three times ultrafiltration against distilled water over a YM3 Ultrafiltration membrane (AMICON®) freezes the remaining residue dried.

3.21 g (98.6% of theory) of the title compound are obtained as an amorphous, colorless Powder with a water content of 7.30%.

Elemental analysis (calculated on anhydrous substance):
calc .: C 41.00; H 5.68; N 9.17; S 4.01; Gd 11.80; Na 1.72;
Found: C 41.09; H 5.71; N 9.10; S 4.08; Gd 11.74; Na 1.74;

The molar element ratio of sulfur to gadolinium results thus an average degree of loading of the polymer of 37.8% Gd-DTPA and 62.2 % α-D-mannose (corresponding to 18 Gd-DTPA units and 30 α-D-mannose residues per molecule). There are no ninhydrin reactions in the title compound free amino functions more detectable.

Example 62 48 cascade polyamine (Gd-DTPA amide) ₁₈ - [3- (1-thio-β-D-glucopyranosyl) propionic acid amide] ₃₀ conjugate a) 48 cascade polyamine- [DTPA-tetra-t-butyl ester amide (terminal)] ₁₈ and [3- (2,3,4,6-tetra-O-acetyl-1-thio-β-D-glucopyranosyl ) -propionate] ₃₀ - conjugate

In an analogous manner as described for Example 61b), the conversion of 3.26 g (0.19 mmol; corresponding to 5.70 mmol of free amine functions) of the Ti tel compound from Example 61a) with 3.3 g (6.3 mmol) of 3- (2,3,4,6-tetra-O-acetyl- 1-thio-β-D-glucopyranosyl) propionic acid N-hydroxysuccinimide ester 5.45 g (94.3 % d. Th.) Of the title compound as a colorless oil.

Elemental analysis:
calc .: C 53.52; H 7.48; N 7.22; S 3.16;
Found: C 53.88; H 7.46; N 7.20; S 3.20;

The title compound shows a negative ninhydrin reaction. From the percent len sulfur value of the elementary analysis results in an average degree of loading of the polymer of 30 β-D-glucose residues per molecule.

b) 48 cascade polyamine (DTPA amide) ₁₈ - [3- (1-thio-β-D-glucopyra nosyl) propionic acid amide] ₃₀ conjugate

In an analogous manner to that described for Example 61c), the deprotection of 5.17 g (0.17 mmol) of the title compound from Example 62a) leads to the formation of 3.35 g (88.0% of theory) after freeze drying. ) the above-mentioned title compound as an amorphous powder.
Water content: 7.44%

Elemental analysis (calculated on anhydrous substance):
calc .: C 43.91; H 6.09; N 9.82; S 4.29;
Found: C 43.94; H 6.11; N 9.77; S 4.30;

c) 48 cascade polyamine (Gd-DTPA amide) ₁₈ - [3- (1-thio-β-D-glucopyranosyl) propionic acid amide] ₃₀ conjugate

In an analogous manner as described for Example 61d), the complexation of 3.04 g (0.136 mmol; corresponding to 2.45 mmol of DTPA) of the title compound from Example 62b) with 556 mg (2.5 mmol) of gadolinium chloride, dissolved in 10 ml of distilled water to form the title compound mentioned above.
Yield: 3.21 g (98.4% of theory) as a colorless and amorphous powder
Water content: 5.03%

Elemental analysis (calculated on anhydrous substance):
calc .: C 41.00; H 5.68; N 9.17; S 4.01; Gd 11.80; Na 1.72;
Found: C 40.96; H 5.75; N 9.20; S 3.97; Gd 11.78; Na 1.69;

Average degree of loading per molecule of the above title compound:
18 Gd-DTPA units (polymer loading level of 43.8%)
and 30 D-glucopyranosyl residues (degree of polymer loading of 56.2%)

The polymer loading levels were calculated from the molar element ratios determined from sulfur to gadolinium.

Example 63 13 kD poly-L-lysine (Gd-DTPA amide) ₄₇ - [1- (4-thioureidophenyl) -N-acetyl-β-D-glucosaminopyranosyl] ₃₃ conjugate a) 13 kD poly-L-lysine (DTPA amide) ₄₇ - [1- (4-thioureidophenyl) -N-acetyl-β-D-glucosaminopyranosyl] ₃₃ conjugate

1.5 g (0.047 mmol; corresponding to 1.54 mmol of free amino functions) 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 2.35 g (6.63 mmol) of p-iso thiocyanatophenyl-N-acetyl-β-D-glucosaminopyranose (representation according to: Jonas Adam J, Jobe Helgi, Biochem. J. (1990), 268 (1), 41-5), where the pH of the reaction solution using 0.1 molar sodium hydroxide solution at 9.5 con is kept constant. After a reaction time of 12 hours at 22 ° C, fil trated and made up to a total volume of 250 ml with distilled water. Ultrafiltration of the reaction solution three times via a YM3 ultrafiltration membrane (AMICON®) followed by freeze-drying the remaining back stands delivers 1.93 g (93.8% of theory) of an amorphous and colorless solid fabric.  

Water content: 7.43%

Elemental analysis (calculated on anhydrous substance):
calc .: C 44.78; H 5.60; N 11.74; S 2.42; Na 9.87;
Found: C 44.82; H 5.63; N 11.70; S 2.37; Na 9.79;

The ninhydrin reaction as well as the TNBS method, which is quantitative as also generally used for the qualitative determination of free amino functions can be placed, are negative, d. H. there are no free ones in the polymer Amino functions. From the percentage sulfur of the Elementarana lysis of the title compound gives a degree of loading of the polymer with 41.6% N-acetylglucosamine (corresponding to 26 N-acetylglucosamine residues per molecule). The colorimetric phenol-sulfuric acid determination to determine the average pyranose loading level of the conjugate gives one as a result average statistical loading of 41.1% N-acetylglucosamine per Molecule and is therefore to be regarded as identical with the degree of loading (33 N- Acetyl-glucosamine units per molecule), which consists of the percentage Sulfur value of the elementary analysis was determined.

b) 13 kD poly-L-lysine (Gd-DTPA amide) ₄₇ - [1- (4-thioureidophenyl) -N-acetyl-β-D-glucosaminopyranosyl] ₃₃ conjugate

1.03 g (0.023 mmol; corresponding to 1.50 mmol of DTPA) of the title compound from Example 63a) are described in an analogy as in Example 1c), with 0.40 g (1.51 mmol) of gadolinium chloride, dissolved in 10 ml of distilled Water, complex. After working up and freeze-drying, 1.48 g (99.4% of theory) of the title compound are obtained as an amorphous powder.
Water content: 0.31%

Elemental analysis (calculated on anhydrous substance):
calc .: C 40.90; H 5.11; N 10.72; S 2.21; Gd 15.41; Na 2.25;
Found: C 40.87; H 5.03; N 10.68; S 2.27; Gd 15.48; Na 2.30;

From the molar elementary ratios of gadolinium to sulfur an average degree of loading of the polymer of 60% Gd-DPTA and 40% N-acetyl-glucosamine (corresponding to 47 Gd-DTPA units and 33 -N-acetyl-β-  D-glucosamine residues per molecule). In the title compound are using ninhydrin reaction no free amino functions detectable.

The T ₁ and T ₂ 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, and
the T ₁ sequence: 180 ° -TI-90 ° [inversion recovery]
and the T ₂ sequence: 90 ° -TE-180 ° [CPMG].

All relaxivity determinations were carried out on a Minispec PC 20. In all cases, a solution of Gd-DTPA with a concentration of 1 mmol / L was used as the reference sample.

R ₁ (water) = 13.8 ± 0.2 (L / mmol.s)
R ₁ (plasma) = 16.1 ± 0.3 (L / mmol.s)
R ₂ (water) = 15.8 ± 0.2 (L / mmol.s)
R ₂ (plasma) = 18.0 ± 0.1 (L / mmol.s)

Example 64 24-cascade polyamine- {10- [7- (4-isothiocyanatophenyl) -2-hydroxy-5-oxo-7- (carboxymethyl) -4-aza-heptyl] -1,4,7-tris (carboxymethyl) -1, 4,7,10-tetraazacyclododecane-gadolinium complex} ₁₂ - [1- (4-thioureidophenyl) -β-D-galactopyranosyl] ₁₂ conjugate a) 24-cascade polyamine- {10- [7- (4-thioureidophenyl) -2-hydroxy-5-oxo 7- (carboxymethyl) -4-aza-heptyl] -1,4,7-tris (carboxymethyl) - 1,4,7,10-tetraazacyclododecane-gadolinium complex} ₁₂ conjugate

At room temperature, 5.0 g (1.06 mmol; corresponding to 25.44 mmol of free amine functions) of 24-cascade polyamine are dissolved in 300 ml of distilled water. The pH of the resulting solution is adjusted by adding this solution dropwise with 1 molar sodium hydroxide solution to a pH of 9.5. Then a total of 15.77 g (20.0 mmol) of the 4-iso-thiocyanatophenyl-DO3A-β-hydroxy-gadolinium complex (representation according to: German Patent Application No. 44 24 923 A1) are added in portions, where at pH -Value of the reaction solution is kept constant at 9.5 by adding 1 molar sodium hydroxide solution. The reaction solution is stirred for 12 hours at room temperature and made up to a total volume of 800 ml by adding 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: 10.2 g (78.6% based on the polyamine used) as an amorphous powder
Water content: 6.94%

Elemental analysis (calculated on anhydrous substance):
calc .: C 45.38; H 5.77; N 12.58; S 3.05; Gd 14.35;
Found: C 45.42; H 5.81; N 13.02; S 2.98; Gd 14.90;

From the content obtained by elemental analysis for gadolinium and sulfur a degree of loading of the polymer with gadolinium results Complexes of 39.7% (corresponding to 12 Gd-DO3A units per molecule). 06762 00070 552 001000280000000200012000285910665100040 0002019728954 00004 06643 After a quantitative ninhydrin reaction, there are still an average of one polymer molecule 12 free amino functions.

b) 24-cascade polyamine- {10- [7- (4-isothiocyanatophenyl) -2-hydroxy-5-oxo-7- (carboxymethyl) -4-aza-heptyl] -1,4,7-tris (carboxymethyl) - 1,4,7,10-tetraazacyclododecane-gadolinium complex} ₁₂ - [1- (4-thioureidophenyl) - β-D-galactopyranosyl] ₁₂ conjugate

In an analogous manner as described for Example 1b), the implementation of 2.0 g (0.16 mmol; corresponding to 1.90 mmol of free amine functions) the title Compound from Example 64a) with 2.65 g (8.36 mmol) of p-isothiocyanatophenyl-β- D-galactopyranose after working up 2.34 g (89.3% of theory) of an amorphous and colorless solid. Water content: 4.97%

Elemental analysis (calculated on anhydrous substance):
calc .: C 46.40; H 5.55; N 11.03; S 4.70; Gd 11.52;
Found: C 46.91; H 5.60; N 11.00; S 4.68; Gd 11.49;

The ninhydrin reaction of the title compound is negative, i. H. the polymer Product does not contain any free amino functions. From the percentage sulfur worth as well as the gadolinium content of the elemental analysis of the title compound the degree of loading of the polymer is 51.2% β-D-galactose (corresponding to 12 β-D-galactose residues per molecule) and a complex degree of loading of the polymer of 48.8% (corresponding to 12 gadolinium com plexing per molecule).

Example 65 48 cascade polyamine (Gd-DTPA amide) ₁₈ - [1- (4-thioureidophenyl) -α- D-mannopyranosyl) ₃₀ conjugate a) 48 cascade polyamine (DTPA amide) ₁₈

A solution of 5.0 g (1.7 mmol; corresponding to 81.7 mmol of free amino functions) 48-cascade polyamine is dissolved in 400 ml of distilled water and a pH of 9.5 is set with 1 molar sodium hydroxide solution. A total of 16.5 g (41.0 mmol) of DTPA monoanhydride monoethyl ester is then added, 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 resulting aqueous product solution 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: 15.63 g (81.4%; based on the polyamine used) as an amorphous powder
Water content: 6.94%

Elemental analysis (calculated on anhydrous substance):
calc .: C 40.82; H 5.26; N 12.64; Na 14.65;
Found: C 40.91; H 5.30; N 12.59; Na 14.58;

After elemental analysis as well as colorimetric titration with 0.0025 molar Gadolinium sulfate standard solution in the presence of xylenol orange as an indicator, the result is a DTPA loading level of the polymer of 38.2% (corresponding 18 DTPA units per molecule). After quantitative ninhydrin reaction as well the quantitative determination of amino groups with the TNBS method are in the Average 30 free amino functions per polymer molecule.

b) 48-cascade polyamine (DTPA amide) ₁₈ - [1- (4-thioureidophenyl) -α- D-mannopyranosyl] ₃₀ conjugate

3.0 g (0.134 mmol; corresponding to 4.03 mmol of free amino functions) of the title compound from Example 65a) are dissolved in 150 ml of deionized water and the pH of the solution is adjusted to 9.5 with 1 molar hydrochloric acid. A total of 5.4 g (17.33 mmol) of p-isothiocyanatophenyl-α-D-mannopyranose is then added in portions, 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 2.45 g (87.9% of theory) of an amorphous and colorless solid.
Water content: 7.48%

Elemental analysis (calculated on anhydrous substance):
calc .: C 44.65; H 5.18; N 8.88; S 4.62; Na 8.39;
Found: C 44.70; H 5.23; N 8.81; S 4.57; Na 8.44;

The ninhydrin reaction as well as the TNBS determination of the title compound are negative, d. H. there are no free amino functions in the polymer. Out the sulfur percentage of the elemental analysis of the title compound there is a degree of loading of the polymer with 61.9% α-D-mannose (corresponding to 30 α-mannose residues per molecule). The colorimetric phenolic Determination of sulfuric acid to determine the mean pyranose load degree of polyamine results in a degree of loading of 62.3% α- D-mannose, d. H. there are 30 α-D-mannose residues per molecule. The so he the average degree of loading of the title compound is identical to the loading degree, which was determined by the percentage element ratios.  

c) 48 cascade polyamine (Gd-DTPA amide) ₁₈ - [1- (4-thioureidophenyl) -α- D-mannopyranosyl] ₃₀ conjugate

2.0 g (0.096 mmol; corresponding to 1.73 mmol of DTPA) of the title compound from Example 64b) are dissolved in 60 ml of sodium citrate buffer (pH 5.3) and at room temperature with a solution of 465 mg (1.76 mmol) of gadolinium chloride in 10 ml of distilled water are added. 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: 2.1 g (98.2% of theory) as an amorphous powder
Water content: 4.55%

Elemental analysis (calculated on anhydrous substance):
calc .: C 41.65; H 4.83; N 8.28; S 4.31; Gd 12.68; Na 1.85;
Found: C 41.72; H 4.88; N 8.24; S 4.30; Gd 12.71; Na 1.79;

From the molar elementary ratios of gadolinium to sulfur thus an average degree of loading of the polymer of 36.5% Gd-DPTA and 63.1% α-D-Mannose (corresponding to 18 Gd-DTPA units and 30-α-D-Man residues per molecule). In the title link are by means of the Ninhydrinreak tion no free amino functions detectable.

Claims (20)

1. conjugates consisting of an amino group-containing polymer, Signaling chelate complex group (s) containing metal ion (s) for the diagnostic imaging and mono- or oligosaccharides. 2. Conjugates according to claim 1, characterized in that they are constructed according to the general formula I.
P (K) _m (LZ) _n (I),
wherein
P for a polymer containing k amino groups - of which (n + m) are each reduced by one hydrogen atom - where (n + m) should be ≦ 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 left and
Z represents a mono- or oligosaccharide reduced by one hydroxyl group. 3. conjugates of general formula I according to claim 2, characterized records that the sum (n + m) of the signaling groups (K) and the mono- bound by L each reduced by one hydroxyl group or polysaccharides (Z) equal to the number k of those contained in the polymer Is amino groups. 4. Conjugates according to claim 3, characterized in that k for the numbers 12, 24, 32, 36, 40, 48, 64 or 80. 5. Conjugates of the general formula I according to claim 2, characterized records that P for a k amino groups, of which (n + m) each by one Hydrogen atom are reduced, containing dendrimer, polylysine, poly peptide, polyallylamine, poly [N- (2-aminoethyl)] methacrylamide or polysaccha rid stands.   6. Conjugates of the general formula I according to claim 2, characterized in that they contain a signaling group K consisting of a chelate complex consisting of a radical of the general formula II, III, IV, V or VI.
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 ₁ -C ₇ -alkyl radical, a phenyl or benzyl radical,
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 -CH ₂ -CH (OH) -M ² - group with R ³ independently of one another in the meaning of R ² or the group -CH ₂ - (CH ₂ ) _o - COOH and M ¹ and M ² each have the meaning of a phenylene radical or a straight-chain, branched, saturated or unsaturated C ₁ -C ₂₀ alkylene chain which is optionally substituted by 1-5 (CH ₂ ) 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 contains 1-2 phenyleneoxy groups, with the proviso that at least two of the radicals R ¹ stand for metal ion equivalents of the elements of the above-mentioned atomic numbers. 7. Conjugates according to claim 6, characterized in that M ¹ for a group
stands, where α indicates the linkage to the rest-CONR ³ and β indicates the linkage to T ¹ . 8. Conjugates according to claim 6, characterized in that M ² for a group
stands, where α indicates the linkage to the rest-CH (OH) - and β the linkage point to T ¹ . 9. Conjugates according to claim 1, characterized in that they are mono saccharide hexoses, pentoses or N-acetyl-neuramino acid, each are in the pyranose or furanose form, or their derivatives ent hold. 10. Conjugates according to claim 1, characterized in that they are mono saccharide hexoses, which are in the pyranose form, or their deri vate included.   11. Conjugates according to claim 9, characterized in that the mono saccharide 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 are oligo saccharide constructed from the monosaccharides according to claim 9 ten. 13. Conjugates of the general formula I according to claim 2, characterized records that Z for a at the C-1 atom reduced by one hydroxyl group Mono- or oligosaccharide. 14. Conjugates of the general formula I according to claim 2, characterized records that Z for a at the C-2 atom reduced by one hydroxyl group Mono- or oligosaccharide. 15. Conjugates of the general formula I according to claim 2, characterized in that L represents a group T ² -M ³ -X, where M ^{3 stands} for a direct bond or has the meaning given for M1, T ² for a - NHCS-, -C (= NH) -, -CH ₂ - or CO group and binds to P, X stands for NH, CO, O and S and binds to Z. 16. Conjugates according to claim 15, characterized in that M ³ for a direct bond or for the groups
stands, where γ indicates the junction at X and δ the junction at T ² . 17. Conjugates according to claim 1, characterized in that not by the central ion partially or completely substituted acidic hydrogen atoms by cations of inorganic and / or organic bases or amino acids or amino acid amides are replaced. 18. 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 polymer containing k amino groups - of which (n + m) are each reduced by one hydrogen atom - where (n + m) should be k,
n and m each for the 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
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 ₁ -C ₇ -alkyl radical, a phenyl or benzyl radical,
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 -CH ₂ -CH (OH) -M ² - group with R ³ independently of one another in the meaning of R ² or the group -CH ₂ - (CH ₂ ) _o - COOH and M ¹ and M ² each in the meaning of a phenylene radical or a straight-chain, branched, saturated or unsaturated C ₁ -C ₂₀ alkylene chain, which is optionally substituted by 1-5 (CH ₂ ) _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 contains 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,
L for a left 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),
  wherein
  K 'for a radical of the general formula II', III ', IV', V 'or 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 ¹ and U have the meaning given above,
  with n 'mono- or oligosaccharides of the general formula VIII
  L * -Z '(VIII),
  wherein
  n '= 1-3 n,
  L * for a group T ^{2 '} -M ^3' -X with
  T ^{2 '} in the meaning of -NCS, -C (= NH) OCH ₃ , -CHO, a -CO-Fg group where
  Fg for -Cl, -F, -OH,
  -N ₃ , O-CO-R ⁴ with R ⁴ meaning a straight-chain or branched C ₁ -C ₇ 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),
  wherein
  L 'has the meaning given for L, but the carboxy groups optionally 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
  wherein
  R ^{1 '} , 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 formulas
  wherein
  m ', R ^1' , R ² , V, T ^{1 '} , U and Fg have the meaning given above, is implemented, the metal ion introduction which may be necessary taking place before or after the splitting off of the protective groups which may be necessary, with the proviso that if R ^{1 '} stands for protective groups, these must be split off before the metal ion is introduced.

19. Use of at least one physiologically compatible conjugate according to claim 1 for the production of diagnostic agents for the NMR diagnostics. 20. Use of at least one physiologically compatible conjugate according to claim 1 for the production of diagnostic agents for the NMR lymphography.