SK1542003A3 - Perfluoroalkyl-containing complexes comprising sugar residues, method for producing the same and use thereof - Google Patents

Abstract

The invention relates to perfluoroalkyl-containing complexes comprising sugar residues of the general formula (I), wherein R represents a mono- or oligosaccharide residue bound in the 1-OH or 1-SH position, Rf represents a perfluorated carbon chain, K is a metal complex and Y and Z are linker groups. The inventive complexes are suitable for use in intravenous lymphography, in tumor diagnosis and for infarct and necrosis imaging.

Description

Technical field

The invention relates to the subjects as claimed in the claims, i.e. perfluoroalkylated metal complexes with sugar residues of the general formula I, their preparation and their use in NMR diagnostics, X-ray diagnostics, radiodiagnostics and radiotherapy, MRT lymphography and blood-pooling agents. The compounds of the invention are particularly suitable for intravenous lymphography, for tumor diagnosis and for imaging of infarction and necrosis.

BACKGROUND OF THE INVENTION

In nuclear magnetic resonance, the most important element after hydrogen is fluorine:

(1) fluorine has a high sensitivity of 83% hydrogen sensitivity;

2) fluorine has only one NMR-active isotope;

3) the resonant frequency of fluorine is close to the resonant frequency of hydrogen, and therefore both fluorine and hydrogen can be measured on the same instrument,

4) fluorine is biologically inert,

(5) Fluorine does not occur in biological material (except teeth) and can therefore be used as a probe or contrast medium without background disturbance.

These properties have led to the fact that fluorine is given much attention in the patent literature concerning diagncsrrky based on nuclear magnetic resonance: it c ^S ' imsging, functional diagnosis and spectroscopy.

In U.S. Patent 4,639,364 (Mallinckrodt) CF ₃ SO ₂ NH ₂ , CF ₃ SO ₂ NH-CH ₂ - (CHOH) ₄ -CH ₂ OH are suggested as contrast agents for ¹⁹ F-imaging as contrast agents.

¹⁹ German patent DE 4203254 (Max Planck-Gesellschart), in which an aniline derivative is proposed, also deals with F-imaging.

CF

CF.

CF.

NHCOCH, F-imaging is also the subject of patent application WO 93/07907 (Mallinckrodt), where phenyl derivatives are claimed as contrast agents

CO ₂ H

Compounds with a substantially simpler structure are also proposed for * ⁹ E-imaging. U.S. Pat. No. 4,586,511 (Children's Hospital Medical Center) mentions perfluorooctyl bromide Cl ₃ (CF ₃ ) -Br, in European patent E? 307863 (Air Products) is perfluoro-15-crown-5-ether

U. S. Patent No. 4,588,279 (University of Cincinnati Children's Hospital Reasearch Foundation) discloses prefluorocarbons such as perfluorocyclononane or perfluorocycl looktane, perfluorinated ethers such as perfluorotetrahydrofuran, or diethers such as perfluorinated propylene glycol diether ^CF 3

F - ^ - - O - CF ₂ CF ₂ CF - O CF ₃

cf ₃

Likewise, for the ¹⁹ L-imaging compounds, compounds listed in WO 94/22368 in the audio unit (Molecular Biosystems), e.g., -O-CO- (CH ₂₎ -R ₆

CH ₃ (CH ₂ ) ₇ -CH = CH- (CH ₂ ) ₇ -CO-o

O-CO- (CH _{2) p} -R

which contain a perfluoro-1H, If-neopentyl group as the fluorinated residue.

Another structural type with extended diagnostic use is disclosed in US Patent No. 5,362,478 (Vivorx) in which a fluorocarbon / polymeric coating combination is claimed for imaging purposes. Perfiuononan and human serum albumin are named. In addition, this combination can be used to utilize the fluorine atom as a probe for local temperature measurement and for determining the oxygen partial pressure.

Perfluorocarbons are also claimed in U.S. Pat. No. 4,586,511 for the determination of oxygen.

In German patent DE 4008179 (Schering), fluorinated benzenesulfonamides of the formula

For NMR diagnostics, compounds containing iodine and fluorine atoms are also claimed as contrast enhancers in WO 94/05335 and WC 94/22368 (both Molecular Biosystems):

Also, a combination of a fluorine-paramagnetic metal ion is claimed for ¹⁹ F-imaging for open-chain complexes in WO 94/22368 (Molecular Biosystems), for example:

what;

and in EP 292 306 (Terumo Kabushiki Kaisha), for example:

'0 ₂ C' 0 ₂ C

R

I .N, 'CO,

WHAT,

Gd

3+ δ

but also for cyclic compounds as listed

EP 628 316 (Terumo Kabushiki Kaisha)

R = - <= ^Η , - <θ - (- CF,

CF,

Combination of fluorine atom also on NMR-spectroscopic ring):

- Rare earth metal is claimed for temperature measurements (DE 4317588, ScheνΛ Z '\ Z ^c ° 2

what;

Ln: rare earth metal: La, Pr, Dy, Eu

While n compounds containing fluorine and iodine atoms, there are no interactions between the two nuclei, fluorine and paramagnetic centers (radicals, metal ions) exhibit an intense interaction resulting in a shortened relaxation time of the fluorinated nucleus. The magnitude of this effect depends on the number of unpaired electrons of the metal ion (Gd ³⁺ > Mn ² > Fe ^; > Cu ²⁺ ) and the distance between the paramagnetic ion and the 'F' atom.

The more unpaired electrons have a metal ion and the closer they are to a fluorine atom, the greater the relaxation time of the fluorine nucleus.

The reduction in relaxation time as a function of the distance from the paramagnetic ion is observable in all nuclei with an odd spin number (thus also in the proton) and therefore the gadolinium compounds find wide application as contrast agents in nuclear spin tomography (Magnevist®, Prohance®, Omniscan®, Dctarem® ). However, the 1 H-MR-imaging ( ¹ H-MRI) measures and uses the proton relaxation time, T ¹ or T ² , i.e., water protons, rather than the fluorine nucleus relaxation time, to display. The quantitative measure to shorten the relaxation time is re-toxicity (liter / mmol.s).

Paramagnetic ion complexes have been used successfully to reduce relaxation times. The following table shows the re-toxicity of some commercial preparations.

table

δ -Reactivity in water [1 / mmol.s, 39 ° C, 0.47 T) δ -Reactivity in plasma [1 / mmol.s, 39 ° C, 0.47 T] Magnevist.RTM 3.8 4.8 Dotarem.RTM 3.5 4.3 Omniscan.RTM 3.8 4.4 Prohance® 3.7 4.9

In these compounds, there are only interactions between the protons and the gadolinium ion. Thus, for these contrast agents a re-toxicity of about 4 [1 / mmol.s] is observed in water.

Also used for MR-imaging are both fluorine compounds for ¹⁹ F-imaging, which utilize a reduced relaxation time of the fluorine core, as well as fluorine-free compounds for which the relaxation time of water protons is determined.

When the perfluorocarbon residue is introduced into the paramagnetic contrast medium, i.e., by combining the properties known hitherto only as suitable for F-imaging compounds, with the compounds used for proton-imaging, the re-toxicity of protons in water surprisingly also increases substantially and reaches 10 -50 [1 / mmol.s], compared to the values of 3.5 to 3.8 [1 / mmol.s] shown in the table for some commercial preparations.

Perfluoroalkylated metal complexes are already known from DE 196 03 033.1. However, these compounds cannot be satisfactorily used in all cases, so there is still a need for contrast media to represent malignant tumors, lymph nodes and necrotic tissue.

Malignant tumors often metastasize in regional lymph nodes, where more nodes may participate. Thus, lymph node metastases were found in about 50-69% of all malignant tumor patients (Elke, Lymphographie, in the book: Frommhold, Stender, Thurn (eds.), Radiologic Diagnostics in Klinik und Praxis, Vol. IV, Thieme Verlag, Stuttgart) , 7 ^h eds., 434-496, (1984)). The diagnosis of metastatic invasion in lymph nodes is of great importance for the therapy and prognosis of malignant diseases. With modern imaging methods (CT, US and MRI), lymphogenic colonies of malignant tumors are poorly recognized, since in most cases only lymph node size can be used as a diagnostic criterion. Therefore, it is not possible to distinguish small metastases in non-enlarged lymph nodes (<2 cm) from lymph node hyperplasia without malignant invasion (Steinkamp et al., Sonographie and Kernspin tomography: Differentialdiagnostics of Lymphknoten-Vergrönder Metering and Lymphknoten Metering Diseases). 33, 158 (1992)).

It is desirable to be able to distinguish lymph nodes with metastatic invasion from hyperplastic lymph nodes using specific contrast media.

Direct X-ray lymphography (injection of an oil suspension of the contrast agent into a prepared lymphatic vessel) is known; however, it is still a rarely used invasive method that is capable of imaging only a few lymphatic flow sites.

In animal experiments, fluorescently labeled dextrans are used experimentally so that lymphatic effluent can be observed after their interstitial administration. All useful markers for imaging of lymphatic pathways and lymph nodes following interstitial / intracutaneous application also have the common feature that they are particulate (for example, particle-like nanoparticle emulsions and suspensions) or large polymers (see also WO 90 / 14346). However, the preparations described so far are not optimal for indirect lymphography due to their lack of local and systemic tolerability and their lack of diagnostic effectiveness.

Because lymph node imaging is crucial for early recognition of metastatic invasion in cancer patients, there is still a great demand for specific contrast agents to diagnose corresponding changes in the lymphatic system.

A high concentration of the contrast agent and its great stability are just as desirable as being as diagnostically relevant as possible enrichment in the lymph at multiple lymphatic sites. The overall burden on the body should be low due to the rapid and complete excretion of the contrast agent. A rapid onset of action is also important for radiological practice, if possible within a few hours after application of the contrast agent. The patient should also be well tolerated.

Last but not least, it is desirable to have lymph-specific contrast agents which, in a single diagnostic examination, will allow both the primary tumor and possible lymph node metastases to be displayed.

Another important field of medicine is derection, localization and monitoring of necroses or heart attacks. Myocardial infarction is not a stationary process but a dynamic process lasting for a long time (weeks to months). The disease takes place in roughly three phases which are not sharply separated but overlap. The first phase, the development of myocardial infarction, involves 24 hours after the heart attack;

in this phase, destruction proceeds as a shock wave (the frontal wave phenomenon) from subendocard to myocardium. The second phase, an already developed heart attack, involves stabilizing the area in which fibrosis occurs as a healing process. The third phase, the healed heart attack, begins after all the destroyed tissue is replaced by fibrous scar tissue. During this period, extensive restructuring takes place.

To date, no precise and reliable method is known to be able to diagnose the actual phase of a myocardial infarction in a living patient. In assessing a heart attack, it is critical to know the extent of the lost tissue and where the loss occurred, as the treatment depends on this information.

Heart attacks occur not only in the myocardium, but also in other tissues, especially the brain.

While infarction is treatable to some extent, necrosis, which is locally restricted by tissue death, can only prevent or mitigate the harmful consequences for the rest of the organism. Necrosis can occur in many ways: injuries, chemicals, oxygen deficiency or radiation. As with a heart attack, knowledge of the extent and nature of necrosis is important for further medical care.

Previously, there have been attempts to improve the localization of infarction and necrosis by using contrast agents in non-invasive methods such as scintigraphy or nuclear spin tmography. There are many attempts in the literature to use porphyrins for imaaing necrosis, but the results are contradictory. Thus, Winkelmann and Hoyes [Nature, 200, 903 (1967)] describe that manganese-5,10,15,20-tetrakis (4-sulfonatophenyl) porphyrin (TPPS) selectively accumulates in the necrotic portion of the tumor.

In contrast, Lyon et al., [Magn. Res. Med. 4, 24 (1987)] observed that manganese-TPPS was dispersed throughout the body into the kidneys, liver, tumor, and only to a minor extent in the muscles. It is interesting to note that the concentration in the tumor reaches a maximum of day 4 only after the authors have increased the dose from 0.12 mmol / kg to 0.2 mmol / kg. The authors therefore also talk about non-specific uptake of TPPS in the tumor. Bockhurst et al. [Acta Neurochir. 60, 347 (1994, Suppl.)] That Mn-TPPS selectively binds to tumor cells.

Ioster et al., [J. Nucl. Med. 26, 756 (1985)] have found that the enrichment ^lu yn-5,10,15,20-tetrakis (4-N-methyl-pyridinium) porphyrin (TMPyP) does not occur in the necrotic portion, but in the living edge layers. While it can be inferred that porphyrin-tissue interaction occurs, this is not proven.

Ni et al. published by [Circulation, Vol. 90, no. 4, vol. 2, p. 1468; Abstract 2512 (1994)] that are capable of imaging infarcted areas with manganese-tetrafenylporphyrin (Mn-TPP) and gadolinium-mesoporphyrin (Gd-MP). In International Patent Application WO 95/31219, both compounds are proposed for imaging infarction and necrosis. The authors, Marchal and Ni, found (Example 3) that for Gd-MP, the metal content of the infarcted tubule was similar to that of a healthy organ, while myocradle was nine times higher for the infarcted tissue (Example 1). Remarkably, the ratio of signal intensities in MRI for infarcted tissue compared to healthy tissue was comparable in both cases (2.10 and 2.19, respectively). Other metalloporphyrins are described in patent application DE 19835082 (Schering AG).

Porphyrins tend to deposit in the skin, leading to photosensitization. This sensitization can take days, but also weeks, and is an adverse effect when using porphyrins for diagnostic purposes. Nevertheless, the therapeutic index for porphyrins is only very small because, for example, for Mn-TPPS, the effective dose is up to 0.2 mmol / kg, while the LD50 is already 0.5 mmol / kg.

Contrast agents for imaging necroses and infarcts not derived from porphyrins are described in DE 19744003 (Schering AG), DE 19744004 (Schering AG) and WO 99/17809 (ΞΡΙΧ). However, to date, there are no compounds that can be used satisfactorily as contrast agents for imaging heart attacks and necroses.

It is therefore an object of the present invention to find contrast agents which are particularly useful for MRT lymphography, but also for the diagnosis of tumors and imaging of necroses and heart attacks. This object is solved in the present invention.

SUMMARY OF THE INVENTION

The invention relates to perfluoroalkylated complexes containing sugar moieties of the general formula I (K) 1 - G - (ZR ^f ) _m

I (I) (Y-R) p wherein

R is a monosaccharide or oligosaccharide residue linked through the 1-OH or 1-SH position,

R ^f is a perfluorinated, straight or branched hydrocarbon chain of the formula -C _n F _2n E, where E is a terminal fluorine, chlorine, bromine, iodine or hydrogen atom, n equals a number from 4 to 30,

K is a metal complex of formula II

CO OR ¹ where

R ¹ is a hydrogen atom or a metal ion equivalent with atomic number 21-29, 31-33, 37-39, 42-44, 49 or 57-83, provided that at least two R ¹ are metal ion equivalents,

R ² and R ³ independently of one another are hydrogen, C 1 -C ₇ -alkyl, benzyl, phenyl, -CH ₂ OH or -CH ₂ OCH 3, and

U is -CsH ^-O-CH?; ^{- (-)} -, - (CH ₂ ) i-5- (O, phenylene, -CR ₂ NHCC-CH ₂ -CH (CH ₂ COOH) -C ₈ H ₄ -co-, -C ₅ K ₄ - (- OCH 2 CH 2) 0-1 -N (CH 2 COOH) -CH ₂ - (O, or C 1 -C ₁₂ -alkylene or C 7 -C 12 -C 8 H ₄ -O-, the latter two groups being optionally interrupted by one or more oxygen atoms, one to three -NHCO- groups, one to three -CONH- groups, and / or are substituted by one to three - (Cb ₂ ) 0-5COOH groups, where ω is the point of attachment to -CO-, or the radical of formula III

where

R ¹ is as defined above, R ⁴ is a hydrogen atom or a metal ion equivalent defined for R ¹ and U ¹ is -C ₆ H -O-CH ₂ -co-, where ω is the point of attachment to -CO-, (IV ) or is a radical of formula IV

R ¹ OOC

wherein R ¹ and R ² are as defined above, or is a radical of formula VA or VB

ROOC '/''' -COOR ¹ '

-N \ _n ^ COOR ^ - COOR ¹ (VA)

ABOUT

II

I / O — C '^^ COOR ¹

Z '- COOR ¹ -N

V ^ COOR ¹ (UK)

-COOR ¹ wherein R ¹ is as defined above, or is a radical of formula VI

ROOC— \ CO— «λ.

r.

ROOC- 'COOR ¹ wherein R ¹ is as defined above, or a radical of formula VII (VI)

ROOC

ROOC

ABOUT

ROOC (VII)

Where R ¹ is as defined above and

The ^X is -ΟβΗ ^ -Ο-ΟΗζ-ω-, ω represents the point of attachment to -CO-, or a radical of the formula VIII

COOR ¹

COOR ¹ wherein R ¹ is as defined above, and optionally free acid groups present in K may optionally exist as salts of organic and / or inorganic bases or amino acids or amino acid amides,

G when K is a metal complex II to VII, is at least three times a functionalized moiety selected from the group of the following moieties a) to j)

a)

β y = CO-CHCH 2 (b) NH 1

zz a

NH-CO-CH-CH-N W-C '

I, N- * Y

H

N-CO-C- (CH 2) ₄ -Nv * -a

NH

I

NH, NH-CO-CH- (CH ₂ ) ₄ -NH 4

g)

N-CO-CH- (CH ₂ ) ₄ -NH-

NH

Y

NH-CO-CH- _(CH2) ^ β -ΝΗ-

NH

-N- (CH ₂ ) ₄ -CN ^H CO ^H

CO-C- (CH ₄ -N-H)

h)

NH β

j) α ^ ΝΗ-CH -CO Y Y β a

G, when K is a metal complex VIII, is at least three times a functionalized moiety selected from k) or 1),

k) p 1 NH- (CH ₂ ) _r CH-CO- v

NH

I

Y

1) α-CO-CH 2 -CH-CH 2 CO 2 Y ² I ²

NH β

where a is the point of attachment of residue G to the complex K, β is the point of attachment of residue G to residue Y and γ is the point of attachment of residue G to residue Z,

Y is -CH 2 -, δ- (CH ₂ ) _η -00-β [where n = 1-5], δ-ΟΗ ₂ -ΟΗΟΗ- (3Ο-β) or δ-CH (CHOH-ClbCH) ) -CHOH-CHOH-CO-β, where δ is the point of attachment to the sugar residue R and β is the point of attachment to the residue G,

Z is a group

y-n

N-SO ₂ -S-yl COCH ₂ -O- _(CH) 2 -SO ₂ -E,

ABOUT

or

7-NH-CH ₂ CH-O-CH ₂ CH ₂ S wherein γ is the point of attachment of Z to radical G, and ε is the point of attachment of Z to the radical R ^{perflučrovanému:} and

1, m independently of each other are an integer of 1 or 2, and p is an integer of 1 to 4.

If the compound of the invention is intended for use in NMR diagnostics, the metal ion in the signal giving group must be paramagnetic. Such ions are especially divalent and trivalent ions of the elements of atomic numbers 21-29, 42, 44 and 58-70. Suitable ions are, for example, chromium (III), iron (II), cobalt (II), nickel (II), copper (II), prazeodymium (III), neodymium (III), samarium (III), and yterbium (III) ions. Gadolinium (III), terbium (III), dysprosium (111), holmium (III), erbium (III), iron (III) and manganese (II) ions are particularly preferred for their strong magnetic moment.

For the use of the compounds of the invention in nuclear medicine (radiodiagnostics and radiotherapy), the metal ion must be radioactive. For example, radioisotopes of atomic number elements are suitable

27, 29, 51-33, 37-39, 43, 49, 62, 64, 70, 75 and 77. Technetium, gallium, indium, rhenium and yttrium are preferred.

If the compound of the invention is to be used in X-ray diagnostics, it is preferable to work with a metal ion of a higher atomic number, so that sufficient X-ray absorption has been achieved. It has been found that diagnostic means which contain a physiologically acceptable complex salt with metal ions with atomic numbers 25, 26, 39 and 57-83 are suitable for this purpose.

Preferred are the manganese (II), iron (II), iron (III), prazeodymium (III), neodymium (III), samarium (III), gadolinium (111), yterbium (III) or bismuth (III) ions, particularly the ions dysprosia (III) and yttria (III).

The acidic hydrogen atoms optionally present in the radical R ^{@ 1} , i.e. those which are not substituted by a central ion, can optionally be wholly or partially exchanged with cations of inorganic and / or organic bases or amino acids or amino acid amides.

Suitable inorganic cations are, for example, lithium ion, potassium ion, calcium ion, and especially sodium ion. Suitable organic base cations are derived, inter alia, from primary, secondary or tertiary amines such as ethanolamine, diethanolamine, morpholine, glucamine, N, N-dimethylglucamine, and especially N-methylglucamine. Suitable amino acid cations are derived, for example, from lysine, arginine and ornithine, as well as amides of otherwise acidic or neutral amino acids.

Particularly preferred compounds of formula I are those containing the macrocycle K of formula II.

The radical U in the metal complex K is preferably -CH 2. ^- or -C 8 H 4 -O-CH ₂ - (ω, where ω is the point of attachment to -CO-.

The alkyl groups R @ ² and R @ ³ in the macrocycle of formula II may be unbranched or branched. They are, for example, methyl, ethyl, propyl, isopropyl, n-butyl, 1-methylpropyl, 2-methylpropyl, n-pentyl, 1-methylbutyl, 2-methylbutyl, 3-methylbutyl or 1,2-dimethylpropyl. Preferably, R ² and R ³ are each independently hydrogen or C 1 -C 4 -alkyl.

In a particularly preferred variant of the invention, R ^{2 is a} methyl group and R ³ is a hydrogen atom.

The benzyl or phenyl group R ² or R ^J in the macrocycle K of formula II can also be substituted in the ring.

The radical R in formula I is a monosaccharide or olicosaccharide or thiosaccharide radical which is bonded via the 1-OH position or the 1-SH position, and according to the invention there may also be c-deoxy sugars which contain a hydrogen atom instead of one or more OH groups. In one preferred variant of the invention, R is a monosaccharide residue of 5 or 6 carbon atoms, preferably a residue of glucose, mannose, galactose, ribose, arabinose or xylose, or a corresponding deoxycucre such as c-decxvalactose (fucose) or 6-deoxymanose (rhamnose) or a corresponding thiocug, with glucose, mannose and galactose being particularly preferred.

Z C of the compounds of formula I of the invention are also preferred are those in which ^Rf is -C _n F _2r. ^ I, wherein n is 4-15 Desirably. Particularly particularly preferred are -C ₄ F 9, -C ₆ F 11, -C ₃ F 17, -C ₁₂ F 15, and -C ₄ F ₁₁ , just as the groups in the compounds described in the Examples.

At least three times the functionalized moiety G in formula I, which represents a backbone, is in one preferred variant of the invention lysine residue (a) or (b).

The radicals Y and Z in the general formula I represent the above-mentioned junction, wherein independently of one another the radical y-n-so _2- is preferred for Z and the radical δ-ΟΗ ₂ ΟΟ-β is preferred for Y.

Perfluoroalkyl metal complexes with sugar radicals of general formula I (K) i - G - (ZR ^f) _m (Ϊ) (YR) p wherein A is a metal complex of general formulas II to VII and G is of formula a) to j), wherein Y , Z, R, R ^f, m, AR 1 are defined as above, are prepared by a manner known per se, a carboxylic acid of the formula IIa

wherein R ⁵ is a metal ion equivalent having atomic number 21-29, 31-33, 37-39, 42-44, 49 or 57-83, or a carboxyl protecting group, and R ² , R ³ and U have the foregoing or a carboxylic acid of the formula IIIa

wherein R ⁴ , R ⁵ and U ¹ are as defined above, or a carboxylic acid of formula IVa

wherein R ⁵ and R ² are defined as above, or a carboxylic acid of formula Va or Vb

-N

HOOC / (Va) (Vb) wherein R ⁵ is as previously defined, or a carboxylic acid of Formula VIa

COOR ⁵

COOR ³

COOR ⁵

COOR ⁵

R ^S OOC

R ^S OOC

CO-OH

COOR ⁵

VIa) wherein R ⁵ is as defined above, or a carboxylic acid of formula VIIa

OH (VHa) wherein R ⁵ and U ¹ have the above meanings, optionally in activated form, are subjected to a condensation reaction with an amine of formula IX

H-G- (ZR ^f ) _m

I (IX) (YR) p wherein G is as defined in formulas (a) to (j) and R, R 1, Y, Z, and m are as previously defined, then optionally protecting groups present are cleaved to form a metal complex of formula I, or when R 'is a protecting group, after cleavage of these protecting groups, they are reacted, in a known manner, with at least one metal oxide or metal salt of atomic number 21-29, 31-33, 37-39, 42- 44, 49 or 57-53, and eventually, if desired, the acidic hydrogen atoms present are replaced by cations of inorganic and / or organic bases, amino acids or amino acid amides.

Compounds of formula (I) according to the invention in which K is a metal complex of formula (VIII) and G is defined by formulas (k) or (1) are prepared by a method known in the art for an amine of formula (IIa)

(VIIIa) wherein R ⁵ is a metal ion equivalent having atomic number 21-29, 31-33, 37-39, 42-44, 49 or 57-83, or a carboxyl protecting group, is subjected to a condensation reaction with a carboxylic acid, optionally activated, of formula X

HO - G - (Z - R ^f ) _m

I (X) (YR) p wherein G is defined by formula k) or 1) and R, R ^f , Y, Z, m have the meanings given above, then optionally protecting groups present are cleaved to give a metal complex of formula I, or when R ⁵ is a protecting group, after cleavage of these protecting groups in a subsequent step, it is reacted in a manner known per se with at least one metal oxide or metal salt of the element number 21-29, 31-33, 37- 39, 42-44, 49 or 57-83, then optionally acidic hydrogen atoms present are replaced with cations of inorganic and / or organic bases, amino acids or amino acid amides as appropriate.

The carboxylic acids IIa to VIIa used are either known compounds or are prepared according to the procedures described in the Examples. For example, the preparation of carboxylic acids of formula IIa is described in DE 196 52 386. Carboxylic acids of formula IIIa can be prepared analogously to Example 3 of this application. Carboxylic acids of the formula IVa can be prepared according to DE 197 28 954.

A precursor for compounds of formula VA is the N ³ - (2,6-dioxomorpholinoethyl) -N 0 - (ethoxycarbonylmethyl) -3,6-diazaoctanedioic acid described in EP 263 059.

Compounds of formula (VB) are derived from isomeric diethylenetriaminepentaacetic acid, which binds through acetic acid at a medium nitrogen atom. This DTPA is described in DE 195 07 819 and DE 195 08 058.

Compounds of formula VI are derived from N- (carboxymethyl) -N- [2- (2,6-dioxo-4-morpholinyl) ethyl] glycine, the preparation of which is described in J. Team. Oil. Chem. Soc. (1982) 59 (2)

104-107.

The compounds of formula VII are derived from 1- (4-carboxymethoxybenzyl) ethylenediaminetetraacetic acid, the preparation of which is described in U.S. Pat. No. 4,622,420.

The preparation of amines of formula (IX) and carboxylic acids of formula (X) is described in detail in the Examples and may be carried out analogously to the procedures described therein. The amine of formula (IIa) is a known compound.

Perbenzylated sugar acids used as starting compounds can be prepared analogously to Lockhoff (Angew.

Chem. 1998, 110, Nr. 24, p. 3634 et seq.). Thus, for example, 1-O-acetic acid is prepared from perbenzylglucose in two stages through trichloroacetimidate and reacted with ethyl hydroxyacetate to catalyze BF ₃ in tetrahydrofuran, followed by saponification with sodium hydroxide in methanol-tetrahydrofuran.

In one more preferred process, the starting perbenzylated sugar acids can also be prepared by dissolving perbenzyl 1-OH-sugars in a water-immiscible organic solvent and reacting with an alkylating agent of formula XI

Nu-L-COO-Sg (XI), where Nu is a nucleophagous group, L is a - (CH 2) _n - (where n = 1-5) group, -CH ₂ -CHOH- group, CH (CHOH-CH group) _And OH represents a protecting group in the presence of a base and optionally a phase transfer catalyst. The nucleofusion group in the alkylating agent of formula XI may be, for example, -Cl, -Br, -I, -OTs, OMs, -OSO ₂ CF _; , A group -OSO ₂ C ₄ F: - or -OSC CSF: ^ ?.

The protecting group is a conventional acid protecting group. These protecting groups are well known to those skilled in the art (see Protective Groups in Organic Syntheses, 2nd Edition, T.W. Greene and P.G.M. Wuts, John Wiley & Sons Inc., New York 1991).

The reaction according to the invention can be carried out at temperatures of 0 to 50 ° C, preferably at 0 ° C to room temperature. The reaction time is 10 minutes to 24 hours, preferably 20 minutes to 12 hours.

The base is added either in solid form, in particular as a fine powder, or in the form of a 10-70%, especially 30-50%, aqueous solution. Preferred bases are NaOH and KOH.

As organic water-immiscible solvents, for example, toluene, benzene, CF ₃ -benzene, hexane, cyclohexane, diethyl ether, tetrahydrofuran, dichloromethane, MTB or mixtures of these solvents can be used in the alkylation process of the invention.

The quaternary ammonium or phosphonium salts as well as crown ethers such as [15] -crown-5 or [18] -crown-6 serve as phase transfer catalysts according to the invention. Preferred are quaternary ammonium salts containing four identical or different hydrocarbon groups selected from the group consisting of methyl, ethyl, propyl, isopropyl, butyl or isobutyl. The hydrocarbon groups in the cation must be large enough to allow good solubility of the alkylating agent in the organic solvent. N (butyl) ₄ ⁺ -C 1, N (butyl) 4 ⁺ -HSO 4, but also N- (methyl) 1 -C 1, are particularly preferred according to the invention.

The metal complexes according to the invention have proved to be particularly suitable for NMR and X-ray diagnostics, but also for radiodiagnostics and radiotherapy. The invention therefore also relates to the use of the perfluoroalkylated metal complexes with sugar residues according to the invention for the preparation of contrast agents for use in NMR and X-ray diagnostics, in particular in lymphography, tumor diagnostics and imaging of infarcts and necroses, as well as in radiodiagnostics and radiotherapy. The compounds of the invention are well suited for use in interstitial and especially intravenous lymphography. In addition, they can also serve to display the vascular system (as blood-pool reagents).

The invention also relates to pharmaceutical compositions comprising at least one physiologically acceptable compound according to the invention, optionally together with galenically customary additives.

The compounds of the present invention are characterized by very good systemic tolerability and high enrichment in lymph nodes at three consecutive sites (which is particularly important for i.v. lymphography). They are therefore particularly suitable for use in MRT lymphography.

The compounds of the invention are also well suited for the recognition and localization of vascular diseases, since they are dispersed exclusively therein when applied to the intravasal space. The compounds according to the invention make it possible, by means of nuclear spin tomography, to distinguish well-perfused tissue from poorly perfused tissue and thereby to diagnose ischemia. Using the contrast agents of the invention, the infarcted tissue can also be distinguished from the surrounding of healthy or ischemic tissue due to its anemia. This is particularly important when, for example, a heart attack is to be distinguished from ischemia.

Compared to macromolecular compounds such as Gd-DTPA-polylysine, which have hitherto been used as a block-pool reagent, the compounds of the invention also exhibit a higher T ¹ re-toxicity and thus have a higher signal intensity in NMR imaging. In addition, since they have longer retention in the bloodstream, they can be administered at relatively low doses (from, for example, <50 μιηοΐ Gd / kg body weight). In particular, however, the compounds of the invention are rapidly and completely excreted from the body.

It has further been found that the compounds of the invention concentrate in areas with increased vascular permeability, such as in tumors, thereby making conclusions about tissue perfusion, allowing the determination of tissue blood volume, selectively shortening relaxation times or blood densities, and displaying vascular permeability. Such physiological information cannot be obtained using extracellular contrast media such as Gd-DTPA (Magnevist®). These areas also imply application areas in the field of modern imaging techniques, nuclear spin tomography and computed tomography: it is a specific diagnosis of malignant tumors, early therapy control for cytostatic, anti-inflammatory or vasodilative therapy, early recognition of poorly perfused areas (eg in myocardium), angiography in vascular diseases and the recognition and diagnosis of sterile or infectious inflammations.

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

If suspensions or solutions of the compositions of the invention in water or physiological saline are desired for enteral or parenteral administration or for other needs, they are mixed with one or more galenically customary excipients (such as methylcellulose, lactose or mannitol) and / or surfactants (such as lecithin, Tween® or Myrj®) and / or flavorings (such as essential oils).

In principle, the pharmaceutical compositions of the invention may also be prepared without isolation of the complexes. In any case, however, care must be taken to ensure that the chelate formation is carried out in such a way that the formed complexes according to the invention contain practically no uncomplexed toxic metal ions.

This can be achieved, for example, by color indicators, such as xylene orange, in control titrations during preparation. The invention therefore also relates to a process for the preparation of complex compounds and their salts. The purification of the isolated complex remains the safest.

When administered in vivo, the compositions of the invention may be administered together with a suitable carrier, such as serum or saline, and other pro-teine such as human serum albumin (HSA).

The compositions of the invention are usually administered parenterally, preferably intravenously. They may also be administered intravascularly or interstitially / intracutaneously, depending on whether vessels or tissues are being examined.

The pharmaceutical compositions according to the invention preferably contain 0.1 pmol - 2 mol kmplex / liter and are generally applied in amounts of 0.0001 - 5 mmol / kg.

The compositions of the invention meet the multilateral requirements for nuclear spin tomography contrast media. They are preferably suitable for increasing the signal intensity after oral or parenteral administration, thereby improving the performance of the image obtained by nuclear spin tomcryria. Furthermore, these compositions exhibit the high efficacy necessary to burden the organism with, as far as possible, only small doses of foreign substances, as well as the good tolerability necessary to maintain the non-invasive nature of the examination.

Due to the good solubility of the compositions of the invention in water and low osmolality, it is possible to prepare highly concentrated solutions in order to maintain the volumetric load of the bloodstream within reasonable ranges and to compensate for dilution with body fluid. In addition, the compositions of the present invention not only exhibit high in vitro stability but also surprisingly high in vivo stability, so that release or exchange in the coupled ion complexes (which are themselves toxic) over a period of time until these new contrast agents are completely eliminated from the body .

In general, the compositions according to the invention are applied as NMR diagnostics in an amount of 0.0001-5 mmol / kg, preferably 0.005-0.5 mmol / kg.

Furthermore, the complex compounds of the invention can be advantageously used as susceptible reagents and as shift reagents in in vivo NMR spectroscopy.

Due to their favorable radioactive properties and the good stability of the complex compounds contained therein, the compositions of the invention are also suitable as radiodiagnostics. Details of such use and dosage are described, for example, in Radiotracers for Medical Applications, CRC-Press, Boca Raton, FL.

The compounds and compositions of the invention can also be applied in positron emission tomography using isotopes that emit positrons, such as ⁴³ Sc, ^4, Sc, ⁵² Fe, ⁵⁵ Co, ¹⁸ Ga and ^CS Y (Heiss, WD, Phelps, ME). : Positron Emission Tomography of the Brain, Springer Verlag Berlin, Heidelberg, New York, 1983).

Surprisingly, the compounds of the invention are also useful for distinguishing malignant and benign tumors in areas without a blood-mczog barrier.

The contrast media of the invention can also be used, after intravenous administration, to quantitatively determine hyperoxia-induced changes in capillary integrity (including acute capillary holes and restoration of normal endothelial integrity after hyperoxic damage).

Histological studies confirm regional microvascular hyperpermeability.

Thus, the contrast agents of the invention can also be used to display abnormal capillary permeability.

These compositions are also characterized in that they are completely excreted from the body and consequently are well tolerated.

Since the compounds of the invention accumulate in malignant tumors (no diffusion into healthy tissue, but high permeability in tumor vessels), they may also promote radiation therapy for malignant tumors. This differs from the corresponding diagnosis only in the amount and nature of the isotope used. The aim is to destroy the tumor cells by high-energy short-wave radiation and with as little dispersion as possible. Interactions in a complex of contained metals (such as iron or gadolinium) with ionizing radiation (e.g. X-rays) or neutrons are utilized for this purpose. As a result of this effect, the local dose of irradiation at the site where the metal complex is located (e.g. in tumors) is substantially increased. To achieve the desired dose of irradiation in malignant tissues, the use of such metal complexes can significantly reduce the radiation load of healthy tissue and thus avoid the side effects of the burden on the patient. The metal complex conjugates of the invention are therefore also suitable as radiosensitizing agents in the radiation therapy of malignant tumors (e.g., the use of Mossbauer effects or in neutron absorption therapy). Suitable β-emitting ions are, for example, ^4c Sc, ^q 'Sc, ⁴⁸ Sc, ² Ca,' ^J Ca and ⁵⁰ Y. Suitable α-emitting ions with low half-lives are, for example, 2 ^µ Bi, ²¹² Bi, ²¹³ Bi and ^2η Βί. with ²¹² Bi being preferred. Suitable photons and electron emitting ions are ¹⁵⁸ Gd, which can be obtained from ¹⁵⁷ Gd by neutron absorption.

When the composition of the invention is for use in a radiation therapy variant proposed by RL Millscm et al. [Nature, Vol. 336, (1988), p. 787], the central ion must be derived from the Mossbauer isotope such as ^{: 7} Fe or * ⁵ Eu.

For in vivo administration, the compositions of the invention may be administered together with a suitable carrier such as serum or saline, and other proteins, such as human serum albumin. The dosage depends on the nature of the cell disorder, the metal ion used and the nature of the imaging method used.

The compositions of the invention are usually administered parenterally, preferably intravenously. However, as mentioned above, they can also be administered intravasally or interstitially / intracutaneously, depending on whether the vessels or tissues are being examined.

The compositions of the invention are particularly useful as X-ray contrast agents, with particular attention being drawn to the fact that no signs of anaphylactic reactions known to iodine-containing contrast agents have been observed in biochemical and pharmacological studies. They are particularly valuable for their favorable absorption properties in the high voltage areas of the lamp for digital subtraction techniques.

As x-ray contrast agents, the compositions according to the invention, in analogy to, for example, meglumine di-isoisoate, are applied in an amount of 0.1-5 mmol / kg, preferably 0.25-1 mmol / kg.

In particular, higher blood concentrations are achieved with the compounds of the invention than with extracellular contrast media. After i.v. application is dispersed only in the intravasal space, which is a decisive advantage over extracellular contrast media.

DETAILED DESCRIPTION OF THE INVENTION

Example 1

a) 2-N-Trifluoroacetyl-6-N-benzyloxycarbonyl-L-lysine

100 c (356.7 mmol) of 6-N-benzyloxycarbonyl-L-lysine were dissolved in a mixture of 1000 ml of trifluoroacetic acid ethyl ester and 500 ml of ethanol, and the resulting solution was stirred at room temperature for 24 hours. After evaporation to dryness, the residue is crystallized from diisopropyl ether to give 128.9 g (96%) of a colorless crystalline powder, m.p. 98.5 ° C.

analysis:

Calculated: C 51.07, H 5.09, N 7.44, F 15.14;

Found: C 51.25%, H 5.18%, N 7.58%, F 15.03%.

b) 2-N-Trifluoroacetyl-6-N-benzyloxycarbonyl-L-lysine- [1- (4-perfluorooctylsulfonyl) piperazine] amide

To 125 g (332.0 mmol) of the title compound of Example 1a and 188.7 g (332.0 mmol) of 1-perfluorooctylsulfonylpiperazine (prepared according to DE 19603033) in 750 mL of tetrahydrofuran at 0 ° C was added 164.2 g ( 0.664 mmol) of EEDQ (ethyl 2-etcxy-1,2-dihydroquinoline-1-carboxylic acid ethyl ester) and stirred overnight at room temperature. After evaporation in vacuo to dryness, the residue is chromatographed on silica gel with dichloromethane-methanol (20: 1). 286.0 g (93%) of a colorless solid are obtained, m.p. 92 ° C.

analysis:

Calculated: C 36.30, H 2.83, N 6.05, F 41.01, S 3.46; Found: C, 36.18; H, 2.94; N, 5.98; F, 40.87; S, 3.40.

c) 6-N-Benzyloxycarbonyl-L-lysine- [1- (4-perfluorooctylsulfonyl) piperazine] amide

To a solution of 280.0 g (302.2 mmol) of the title compound of Example 1b in 2000 mL of ethanol was added ammonia gas for one hour at 0 ° C. The mixture was stirred at 0 ° C for 4 hours, evaporated to dryness and the residue was triturated with water. The solid was filtered off and dried under vacuum at 50 ° C. Yield 243.5 g (97%) of an amorphous solid.

analysis:

H, 3.28; N, 6.75; F, 38.89; S, 3.86; Found: C 37.55, H 3.33, N 6.68, F 38.78, S 3.81.

d) 6-N-Benzyloxycarbonyl-2-N- [1-O-α-D-carbonylmethyl- (2,3,4,6-tetra-O-benzylmanopyranose) -L-lysine- [1- (4-perfluorooctylsulfonyl) piperazine] amide

To a solution of 100.0 g (120.4 mmol) of the title compound of Example 1c, 72.1 g (120.4 mmol) of 1-O-α-D-carboxymethyl- (2,3,4,6-tetra37)

Of O-benzylmanopyranose and 13.86 g (120.4 mmol) of N-hydroxysuccinimide in 500 ml of dimethylformamide at 0 ° C are added 41.27 g (200 mmol) of N, N-dicyclohexylcarbodiimide. The mixture was stirred at 0 ° C for 3 hours and then at room temperature overnight. The precipitated urea is filtered off, the filtrate is evaporated to dryness in vacuo and the residue is chromatographed on silica gel with dichloromethane-ethanol (20: 1) to give 136.1 g (87%) of the product as a viscous oil.

analysis:

Calculated: C 57.32, H 4.89, N 4.31, F 24.86, S 2.47;

Found: C 57.38, H 5.07, N 4.22, F 24.78, S 2.39.

e) 2-N- [1-O-α-D-carbonylmethyl-mannopyranose] -L-lysine- [1- (4-perfluorooctylsulfonyl) piperazine] amide

To a solution of 130.0 g (100.0 mmol) of the title compound of Example 1d in 2000 mL of ethanol was added 10.0 g of palladium catalyst (10% Pd / C) and the mixture was hydrogenated at room temperature for 12 hours. The catalyst was filtered off and the filtrate was evaporated to dryness in vacuo to give 91.7 g (100%) of a colorless solid.

analysis:

Calculated: C 34.07, H 3.63, N 6.11, S 3.50, F 35.24; Found: C, 33.91; H, 3.72; N, 6.04; S, 3.40; F, 35.31.

f) 6-N- [1,4,7-Tris (carboxylatomethyl) -1,4,7,10-terraazacyclododecane-10- (pentanoyl-3-aza-4-oxo-5-methyl-5-yl)) -2-N- [1-Oa-D-carbonylmethyl-mancpyranose] -L-lysine- [1- (4-perfluorooctylsulfonyl) piperazine] amide, gadolinium complex

50.0 g (54.55 mmol) of the title compound of Example 1e,

6.28 g (54.55 mmol) of N-hydroxysuccinimide, 4.62 g (109.0 mmol) of lithium chloride and 34.35 g (54.55 mmol) of the 1,4,7-tris (carboxylatomethyl) gadolinium complex - 10- (Carboxy-3-aza-4-oxo-5-methylpent-5-yl) -1,4,7,10-tetraazacyclododecane was dissolved in 400 mL of dimethyl sulfoxide with gentle warming. 15.88 g (81.8 mmol) of N, N-dicyclohexylcarbodiimide are then added at 10 ° C and the mixture is stirred overnight at room temperature. The solution was poured into 3000 mL of acetone and stirred for 10 minutes. The precipitated solid is filtered off with suction and purified by chromatography on a RP-18 phase in a water-ethanol-acetonitrile (gradient) system. 75.9 g (91.0%) of a colorless solid are obtained, containing 8.6% water.

Analysis (calculated on the anhydrous substance):

Calculated: C 35.34, H 4.09, N 8.24, S 2.10, F 21.12, Gd 10.28;

Found: C 35.28, H 4.15, N 8.19, S 2.15, F 21.03,

10.14% Gd.

Example 2

a) 6-N- [1,4,7-tris (carboxylatomethyl) -1,4,7,10-tetraazacyclododecane-10-Ν- (pentanoyl-3-aza-4-o-5-yl)] - 2 -N- [1-Oa-D-carbonylmethyl-1-manopyranose and] -L-lysine- [1- (4-perfluoroctylsulfonyl) piperazine] amide, gadolinium complex

50.0 g (54.55 mmol) of the title compound of Example I,

6.28 g (54.55 mmol) of N-hydroxysuccinimide, 4.62 g (109.0 mmol) of lithium chloride and 34.35 g (54.55 mmol) of the gadolinium complex 1,4,7-tris (carboxylatomethyl) -10- (carboxy-3-aza-4-oxo-5-methylpent-5-yl) -1,4,7,10-tetraazacyclododecane was dissolved in 400 mL of dimethyl sulfoxide with gentle heating. 16.88 g (81.8 mmol) of N, N-dicyclohexylcarbodiimide are added at 10 ° C and the mixture is stirred overnight at room temperature. The solution was then poured into 3000 ml of acetone and stirred for 10 minutes. The precipitated solid is filtered off and purified by chromatography on RP-18 phase in a cda-ethanol-acetonitrile gradient. 76.0 g (92.0%) of a colorless solid are obtained, containing 6.88% water.

Analysis (calculated on the anhydrous substance):

Calculated: C 34.90, H 3.93, N 8.32, S 2.12, F 21.33,

10.38% Gd;

Found: C 34.81, H 4.02, N 8.27, S 2.09, F 21.22,

10.19% Gd.

Example 3

a) 2- [4- (Ethoxycarbonylmethyloxy)] phenylacetic acid methyl ester

To 200 g (1204.0 mmol) of 4-hydroxyphenylacetic acid methyl ester and 212.0 g (2000.0 mmol) of sodium carbonate in 2000 ml of acetone are added 233.8 g (1400.0 mmol) of ethyl bromoacetate and the mixture is boiled. 5 hours under reflux. The solid was filtered off and the filtrate was evaporated to dryness in vacuo. The residue was chromatographed on silica gel with n-hexane-ethyl acetate - (15: 1) to give 288.5 g (95.0%) of a colorless oil.

analysis:

Calculated: C 61.90, H 6.39;

Found:% C, 61.75;% H, 6.51.

b) 2- (4-Ethoxycarbonylmethyloxy) phenyl-2-bromoacetic acid methyl ester

To a solution of 285.0 g (1130.0 mmol) of the title compound of Example 3a in 2000 mL of tetrahydrofuran was added 201.0 g (1130.0 mmol) of N-bromosuccinimide and 100.0 mg of dibenzoyl peroxide and the mixture was refluxed for 8 hours. condenser. After cooling in an ice bath, the precipitated succinimide is filtered off, the filtrate is evaporated to dryness in vacuo and the residue is purified by chromatography on silica gel, eluting with n-hexane-acetone (15: 1). 359.2 g (96.0%) of a colorless viscous oil are obtained.

analysis:

Calculated: C 47.28, H 4.57, Br 24.16;

Found: C, 47.19; H, 4.71; Br, 24.05.

c) 2- (4-Ethoxycarbonylmethyloxy) phenyl-2- [1- (1,4,7,10-tetraazacyclododecan-1-yl)] acetic acid methyl ester

To a solution of 603.0 g (3500.0 mmol) of 1,4,7,10-tetraazacyclododecane in 6000 mL of chloroform was added 350.0 g (1057.0 mmol) of the title compound of Example 3b and stirred overnight at room temperature. . The mixture is extracted three times with 3000 ml of water, the organic phase is dried over magnesium sulphate and evaporated to dryness in vacuo. The residue was used in the next reaction without further purification (Example 3d). Yield 448.0 g (100%) of a viscous oil.

analysis:

H, 8.11; N, 13.26.

Found:% C, 59.58;% H, 8.20;% N, 13.18.

d) 2- [4- (carboxymethyloxy)] phenyl-2- [1,4,7-tris- (carboxymethyl) -1,4,7,10-tetraazacyclododecan-10-yl] acetic acid

A solution of 445.0 g (1053.0 mmol) of the title compound of Example 3c and 496.0 g (5270.0 mmol) of chloroacetic acid in 4000 mL of water was adjusted to pH 10 with 30% aqueous NaOH and stirred at 70 ° for 8 hours. C. It is then adjusted to pH 13 with 30% aqueous NaOH and refluxed for 30 minutes. The solution is cooled in an ice bath and adjusted to pH 1 with concentrated hydrochloric acid. After evaporation in vacuo to dryness, the residue is taken up in 4000 ml of methanol and stirred for one hour at room temperature. The precipitated sodium chloride is filtered off, the filtrate is evaporated to dryness and the residue is purified by chromatography on RP-18 phase in a water-ethanol-acetonitrile (gradient) system. 403.0 g (69.0%) of a colorless solid are obtained, containing 10.2% water.

Analysis (calculated on the anhydrous substance):

H, 6.18; N, 10.10.

Found:% C, 51.80;% H, 6.31;% N, 10.01.

e) 2- (4- (carboxymethyloxy)] phenyl-2- [1,4,7-tris- (carboxylatomethyl) -1,4,7,10-tetraazacyclododecan-10-yl] acetic acid, gadolinium complex

To 400 g (721.3 mmol) of the title compound of Example 3d in 2000 mL of water was added 130.73 g (360.65 mmol) of gadolinium oxide and the mixture was stirred at 80 ° C for 5 hours. After filtration, the filtrate is lyophilized. Yield 511 g (100%) of an amorphous solid containing 11.0% water.

Analysis (calculated on the anhydrous substance):

Calculated: C 40.67, H 4.41, N 7.98, Gd 22.19;

Found: C 40.51, H 4.52, N 8.03, Gd 22.05.

f) 6-N- [2- [4- (3-oxapropionyl) phenyl] -2- [1,4,7-tris (carboxylatomethyl) -1,4,7,10-tetraazacyclododecan-10-yl] acetic acid ]] - 2-N- (1-Oa-D-carbonylmethyl-manopyranose) -L-lysine- [1- (4-perfluorooctylsulfonyl) piperazine] amide, gadolinium complex, sodium salt

50.0 g (54.55 mmol) of the title compound of Example 1e,

6.28 g (54.55 mmol) of N-hydroxysuccinimide, 4.62 g (109.0 mmol) of lithium chloride and 38.66 g (54.55 mmol) of the title compound of Example 3e are dissolved in 400 ml with slight heating. DMSO. 16.88 g (81.8 mmol) of N, N-dicyclohexylcarbodiimide are then added at 10 ° C and stirred overnight at room temperature. The solution was poured into 3000 mL of acetone and stirred for 10 minutes. The precipitated solid was filtered off and purified by chromatography on RP-18 in a water-ethanol-acetonitrile (gradient) system. The product obtained is dissolved in a small amount of water, adjusted to pK 7.4 by addition of aqueous sodium hydroxide, and subjected to lycfilization. This gave 79.1 g (89%) of a colorless solid containing 10.3% water.

Analysis (calculated on the anhydrous substance):

Calculated: C 36.86, H 3.77, N 6.88, S 1.97, F 19.82, Gd 9.65;

found; C, 36.75; H, 3.84; N, 6.80; S, 2.03; F, 19.75; C-d, 9.57.

Example 4

a) 6-N- [1,4,7-Tris (tert-butyloxycarbonylmethyl) -1,4,7,10-thesraazacyclododecane-10-carbonylmethyl] -2-N- (1-Oa-D-carbonylmethyl-mannopyranose) -L -lysine- [1- (4-perfluorooctylsulfonyl) piperazine] amide

To a solution of 15.0 g (26.19 mmol) of 1,4,7-tris (tert-butyloxycarbonylmethyl) -10-carboxymethyl-1,4,7,10-tetraazacyclododecane (prepared according to WO 91/05762), 24.0 g (26.19 mmol) of the title compound of Example 1e and 3.01 g (26.19 mmol) of N-hydroxysuccinimide in 150 ml of dimethylformamide are added at 0 ° C 8.25 g (40.0 mmol) of N, N- of dicyclohexylcarbodiimide and stirred overnight at room temperature. The precipitated urea is filtered off and the filtrate is evaporated to dryness in vacuo. The residue was chromatographed on silica gel with dichloromethane-methanol (20: 1). 35.45 g (92.0%) of the product are obtained as a colorless solid.

analysis;

Calculated: C 44.08, H 5.69, N 7.62, F 21.95, S 2.13; Found: C, 44.01, H, 5.81, N, 7.53, F, 21.87, S, 2.03.

b) 6-N- [1,4,7-Tris (carboxylatomethyl) -1,4,7,10-tetraazacyclecadane-10-carbonylmethyl] -2-N- (10aD-carbonylmethyl-manopyranose) -L-lysine - [ 1- (4-perfluorooctylsulfonyl) piperazine] amide, gadolinium complex

To a solution of 30.0 g (20.39 mmol) of the title compound of Example 4a in 50 mL of chloroform was added 300 mL of trifluoroacetic acid. The mixture is stirred for 10 minutes at room temperature, then evaporated to dryness in vacuo and the residue is dissolved in 300 ml of water. 3.69 g (10.19 mmol) of gadolinium oxide are added and stirred at 80 ° C for 5 hours. The solution is evaporated to dryness in vacuo and the product is purified by chromatography on silica gel (water / ethanol / acetonitrile). 11.0 g (37.0%) of a colorless amorphous solid are obtained, containing 11.3% water.

Analysis (calculated on the anhydrous substance):

Calculated: C 34.62, H 3.87, N 7.69, F 22.16, S 2.20, Gd 10.97;

Found: C 34.57, H 3.95, M 7.60, F 22.05, S 2.13, Gd 10.90.

Example 5

a) 6-N- [3,6,9-Tris (carboxymethyl) -3,6,9-triazaundecanedioic acid-1-carboxy-11-oyl] -2-N- [1-O-α-carbonylmethyl-manopyranose 1-L-lysine- [1- (4-perfluorooctylsulfonyl) piperazine] amide

To a solution of 24.0 g (26.19 mmol) of the title compound of Example 1e in a mixture of 100 mL of dimethylformamide and 30 mL of pyridine was added 12.10 g (30.0 mmol) of 3-N- (2,6-dioxomorpholinoethyl) - 6-N- (ethoxycarbonylmethyl) -3,6-diazaoctanedioic acid and stirred at 50 ° C for 5 hours. The mixture was then evaporated to dryness in vacuo. The residue is dissolved in 200 ml of water, the pH of the solution is adjusted to 13 by addition of 20% aqueous sodium hydroxide and stirred for 8 hours at 22 ° C and pH 13. The solution is then adjusted to pH 7.2 by addition of concentrated hydrochloric acid and evaporated under vacuum to dryness. The residue is chromatographed on RP-18 silica gel with water-ethanol-acetonitrile (gradient). 17.26 g (51.0%) of a colorless solid are obtained, containing 9.3% water.

Analysis (calculated on the anhydrous substance):

Calculated: C 37.19, H 4.21, N 7.59, F 25.00, S 2.48; Found: C 37.10, H 4.30, N 7.48, F 25.07, S 2.42.

b) 6-N- [3,6,9-Tris (carboxylatomethyl) -3,6,9-triazaundecanedioic acid-1-carboxy-11-oyl] -2-N- (1-Oa-D-carbonylmethyl-manopyranose) - L-lysine- [1- (4-perfluorooctylsulfonyl) piperazine] amide, gadolinium complex, sodium salt

To 10.0 g (7.74 mmol) of the title compound of Example 5a in 100 mL of water was added 1.40 g (3.87 mmol) of gadolinium oxide and the mixture was stirred at 70 ° C for 2 h. The solution is filtered, the filtrate is adjusted to pH 7.4 by addition of 2N sodium hydroxide and lyophilized. 11.36 g (100%) of an amorphous solid are obtained, containing 10.5% water.

Analysis (calculated on the anhydrous substance):

Calculated: C 32.72, H 3.43, N 6.68, S 2.18, Gd 10.71, Na 1.57, F 22.00;

Found: C, 32.65; H, 3.51; N, 6.71; S, 2.08; Gd, 10.61; Na, 1.68; F, 21.87.

Example 6

a) 6-N-Benzyloxycarbonyl-2-N- [1,4,7-tris (carboxylatomethyl) -1,4,7,10-tetraazacyclododecane] -10- (pentanoyl-3-aza-4-oxo-5) -methyl-5-yl)] - L-lysine- [1- (4-perfluorooctylsulfonyl) piperazine] amide, gadolinium complex

50.0 g (60.20 mmol) of the title compound of Example 1c, 6.93 g (60.20 mmol) of N-hydroxysuccinimide, 5.09 g (120, C mmol) of lithium chloride and 37.91 g (60, 20 mmol) of 1,4,7-tris (carboxylatomethyl) -10 - [(pentanoyl-3-aza-4-oxo-5-ethyl-5-yl)] - 1,4,7,10-tetracyclododecane gadolinium complex is dissolved in 400 ml of dimethylsulfoxide with gentle heating. 20.63 g (100.0 mmol) of N, N-dicyclohexylcarbodiimide are then added at 10 ° C and stirred overnight at room temperature. The solution was poured into 3000 mL of acetone and stirred for 10 minutes. The precipitated solid is filtered off with suction and purified by chromatography on RP-18 silica gel (water / ethanol / acetonitrile). 75.53 g (87%) of a colorless solid are obtained, containing 10.1% water.

Analysis (calculated on the anhydrous substance):

Calculated: C 37.48, H 3.84, N 8.74, S 2.22, F 22.39, Gd 10.90;

Found: C, 37.39; H, 4.02; N, 8.70; S, 2.16; F, 22.29; Gd, 10.75.

b) 2-N- [1,4,7-Tris (carboxylatomethyl)] - 1,4,7,10-tetraazacyclododecane-Gd complex, 10- (pentanoyl-3-aza-4-oxo-5-methyl- 5-yl) -L-lysine- [1- (4-perfluorooctylsulfonyl) -piperazine] amide

70.0 g (48.53 mmol) of the title compound of Example 1d are dissolved in a mixture of 500 ml of water and 100 ml of ethanol, 5.0 g of palladium catalyst (10% Pd / C) are added and subjected to hydrocation at room temperature and pressure of 1 atm when hydrogen is absorbed. The catalyst was filtered off with suction, washed well with ethanol (2 x 75 ml) and the filtrate was evaporated to dryness in vacuo. The product is obtained in the form of a strongly viscous colorless oil containing 9.8% water. Yield 63.5 g (100%).

Analysis (calculated on the anhydrous substance):

Calculated: C 37.48, H 3.84, N 8.74, S 2.22, F 22.39, Gd 10.90;

Found: C 37.39, K 4.03, N 8.65, S 2.20, F 22.31, Gd 10.78.

c) 6-N- (10α-D-carbonylmethyl-2,3,4,6-tetra-O-benzyl-manopyranose) -2-N- [1,4,7-tris (carboxylatomethyl)] - 1,4,7 10-tetraazacyclododecane], Gd-complex-10- (pentanoyl-3-aza-4-oxo-5-methyl-5-yl)] - L-lysine- [1- (4-perfluorooctylsulfonyl) piperazine] amide

50.0 g (38.22 mmol) of the title compound of Example 6b, 4.40 g (38.22 mmol) of N-hydroxysuccinimide, 3.39 g (80.0 mmol) of lithium chloride and 22.88 g (38, 22 mmol) 1-OaD-carboxymethyl-2,3,6,6-tetra-O-benzyl-manopyranose was dissolved in 400 ml of dimethylsulfoxide with gentle heating (30-40 ° C). 10.32 g (50.0 mmol) of N, N-dicyclohexylcarbodiimide are then added at 10 ° C and stirred overnight at room temperature. The solution was poured into 3000 mL of acetone and stirred for 10 minutes. The precipitated solid is filtered off with suction and purified by chromatography on RP-18 silica gel with 46 ethanol / acetonitrile (gradient). 64.25 g (89.0%) of a colorless solid are obtained, containing 10.9% water.

Analysis (calculated on the anhydrous substance):

Calculated: C 46.42, H 4.54, N 6.67, S 1.70, F 17.10, Gd 8.33;

Found: C, 46.36; H, 4.71; N, 6.60; S, 1.61; F, 17.19; Gd, 8.21.

d) 6-N- (10aD-Carbonylmethyl-2,3,4,6-tetra-O-benzyl-manopyranose) -2-N- [1,4,7-tris (carboxylatomethyl) -1,4,7, 10-tetraazacyclododecane], Gd-complex-10- (pentanoyl-3-aza-4-oxo-5-methyl-5-yl)] - 1 '-

-lysine- [1- (4-perfluorooctylsulfonyl) piperazine] amide

60.0 g (31.77 mmol) of the title compound of Example 6c are dissolved in 500 ml of ethanol, 6.0 g of palladium catalyst (10% Pd / C) are added, and hydrogenated at room temperature and 1 atm until it absorbs hydrogen. The catalyst was filtered off with suction, washed well with ethanol (2 x 150 ml) and the filtrate was evaporated to dryness in vacuo. The product was obtained as a colorless solid containing 3.9% water. Yield 48.55 g (100%).

Analysis (calculated on the anhydrous substance):

Calculated: C 35.37, H 4.02, N 8.25, S 2.10, F 21.13, Gd 10.29;

Found: C 35.28, H 4.13, N 8.17, S 2.03, F 21.05, Gd 10.20.

Example 7

a) 1,7-Bis (benzyloxycarbonyl) -4- (2- (N-ethyl-N-perfluorooctylsulfonyl)) amino] acetyl] -1,4,7,10-tetraazacyclododecane

To 50.0 g (113.5 mmol) of 1,7-bis (benzyloxycarbonyl) -1,4,7,10-tetraazacyclododecane and 66.42 g (113.5 mmol) of 2- (N-ethyl-N-) acid perfluorooctylsulfonyl) aminoacetic acid (prepared according to

DE 196 03 033) in 300 ml of tetrahydrofuran at 0 ° C is added 49.46 g (200.0 mmol) of EEDQ (ethyl 2-ethoxy-1,2-dihydroquinoline-1-carboxylic acid ethyl ester) and the mixture is stirred at room temperature during the night. It is evaporated to dryness in vacuo and the residue is chromatographed on silica gel with dichloromethane-methanol (20: 1). Yield: 65.2 g (57%) as a colorless solid.

analysis:

Calculated: C 42.91, H 3.80, N 6.95, F 32.05, S 3.18; Found: C 42.85, H 3.90, N 6.87, F 31.98, S 3.15.

b) 1,7-Bis (benzyloxy) -4- [2- (N-ethyl-N-perfluorooctlsulfonyl) amino] acetyl-10- [10α-D-carbonylmethyl-2,3,4,6-tetra-O-benzyl- mannopyranose] -1,4,7,10 tetraazacyclododecane

To 60.0 g (59.53 mmol) of the title compound of Example 7a and 35.64 g (59.53 mmol) of 10aD-carboxymethyl-2,3,4,6-tetra-O-benzyl-manopyranose (prepared according to DE) 19728954) in 300 mL of tetrahydrofuran at 0 ° C was added 24.73 g (100 mmol) of EEDQ (2-ethoxy-1,2-dihydroquinoline-1-carboxylic acid ethyl ester) and the mixture was stirred at room temperature overnight. It is evaporated to dryness in vacuo and the residue is chromatographed on silica gel with dichloromethane-methanol (20: 1). 76.6 g (81.0%) of product are obtained as a colorless solid.

analysis:

Calculated: C 54.44, H 4.70, N 4.41, F 20.33, S 2.02; Found: C 54.37, H 4.81, N 4.35, F 20.27, S 1.96.

c) 1- [2- (N-Ethyl-N-perfluorooctylsulfonyl) amino] acetyl-7- (1-O-α-D-carbonylmethyl-manopyranose) -1,4,7,10-tetraazacyclododecane g (44.07 mmol) of the resulting compound of Example 7b is dissolved in 800 ml of ethanol, 8 g of palladium catalyst (10% Pd / C) are added, and subjected to hydrogenation at room temperature.

The catalyst is then filtered off with suction and the filtrate is evaporated to dryness in vacuo. The product is obtained as a colorless solid. Yield 42.3 g (100%).

analysis:

H, 3.99; N, 7.30; F, 33.65; S, 3.34. Found: C 35.15, H 4.13, N 7.13, F 33.48, S 3.26.

d) 1,7-Bis [1,4,7-tris (carboxylatomethyl) -1,4,7,10-tetraazacyclodocecane-Gd complex-10- (pentanoyl-3-aza-4-oxo-5-methyl- 5-yl) -4- [2- (N-ethyl-N-perfluorooctylsulfonyl) amino] acetyl-10- (1-Oa-D-carbonylmethyl-manopyranose) -1,4,7,10-tetraazacyclododecene g (20.84 mmol) of the title compound of Example 7c, 5.09 g (120 mmol) of lithium chloride and 37.78 g (60 mmol) of the 1,4,7-tris (carboxylatomethyl) -10-pentanoyl-3-aza-4-oxo gadolinium complex -5-methyl-5-yl) -1,4,7,10-tetraazacyclododecane was dissolved in 400 mL of dimethylsulfoxide with gentle heating. Then 29.67 g (120 mmol) of EEDQ are added at 10 ° C and stirred overnight at room temperature. The solution was poured into 3000 mL of acetone and stirred for 10 minutes. The precipitated solid is filtered off with suction and purified by chromatography on RP-18 silica gel with water-ethanol-acetonitrile (gradient). 13.2 g (29.0%) of a colorless solid are obtained, containing 11.8% water.

Analysis (calculated on the anhydrous substance):

Calculated: C 36.31, H 4.34, N 9.62, S 1.47, F 14.79, Gd 14.41%;

Found: C, 36.24; H, 4.27; N, 9.58; S, 1.51; F, 14.85; Gd, 14.25.

Example 8

a) 1,7-Bis (benzyloxycarbonyl) -4- [2- (N-ethyl-N-perfluorooctylsulfonyl) amino] acetyl-10- [pentanoyl-3-aza-4-oxo-5-methyl-5-yl- [1,4,7-tris (carboxylatomethyl) -Gd-complex-1,4,7,10-tetraazacyclododec49 kan-10-yl] -1,4,7,10-tetraazacyclododecane

50.0 g (49.61 mmol) of the title compound of Example 7a, 5.71 g (49.61 mmol) of N-hydroxysuccinimide, 4.24 g (100 mmol) of lithium chloride and 31.24 g (49.61 mmol) gadolinium complex

1,4,4-Tris (carboxylatomethyl) -10- (pentanoyl-3-aza-4-oxo-5-methyl-5-yl) -1,4,7,10-tetraazacyclododecane is dissolved in 350 with gentle heating ml of dimethylsulfoxide. Then 15.47 g (75 mmol) of N, N-dicyclohexylcarbodiimide are added at 10 ° C and stirred overnight at room temperature. The solution is poured into 2000 ml of acetone and stirred for 10 minutes. The precipitated solid was filtered off with suction and purified by chromatography on RP-18 silica gel with water-ethanol-acetonitrile (gradient). 65.1 g (81.0%) of a colorless solid are obtained, containing 7.9% water.

Analysis (calculated on the anhydrous substance):

Calculated: C 40.79, H 4.11, N 8.65, S 1.98, F 19.94, Gd 9.72;

Found: C 40.71, H 4.20, N 8.58, S 2.03, F 19.87, Gd 9.68.

b) 1- [2- (N-Ethyl-N-perfluorooctylsulfonyl) amino] acetyl-7- (pentanoyl-3-aza-4-oxo-5-methyl-5-yl- [tris (carboxylatomethyl) -1,4] , 7,10-tetraazacyclododecan-10-yl] -Gd complex} -1,4,7,10-tetraazacyclododecane

60.0 g (37.05 mmol) of the title compound of Example 8a are dissolved in 600 ml of ethanol, 6.0 g of a palladium catalyst (10% Pd / C) are added and subjected to hydrogenation at room temperature. The catalyst is then filtered off with suction and the filtrate is evaporated to dryness in vacuo. The product is obtained as a colorless solid containing 3.9% water. Yield 50.06 g (100%).

Analysis (calculated on the anhydrous substance):

H, 4.03; N, 10.37; S, 2.37; F, 23.90.

II, 64% Gd;

Found: C 34.58, H 4.15, N 10.28, S 2.30, F 23.84, Gd 11.57%.

c) 1- [2- (N-Ethyl-N-perfluorooctylsulfonyl) amino] acetyl-4,10-di [1-Oa-D-carbonylmethyl-2,3,4,6-tetra-O-benzyl-manopyranose] - 2- (pentanoyl-3-aza-4-oxo-5-methyl-5-yl- [1,4,7-tris- (carboxylactomethyl) -1,4,7,10-tetraazacyclododecan-10-yl] -Gd (complex) -1,4,7,10-tetraazacyclododecane

40.0 g (29.60 mmol) of the title compound of Example 8b, 2.54 g (60.0 mmol) of lithium chloride and 44.9 g (75.0 mmol) of 1-O-α-D-carboxymethyl-2 3,4,6-tetra-O-benzyl-manopyranose was dissolved in 300 ml of dimethylsulfoxide with gentle heating. Then, 24.73 g (100.0 mmol) of EEDQ are added at 10 ° C and stirred overnight at room temperature. The solution was poured into 3000 mL of acetone and stirred for 10 minutes. The precipitated solid was filtered off with suction and purified by chromatography on RP-18 silica gel with water-ethanol-acezonirrile (gradient). 31.98 g (43.0%) of a colorless solid are obtained; substances containing 3,5% water.

Analysis (calculated on the anhydrous substance):

Calculated: C 53.06, H 5.05, N 5.57, S 1.28, F 12.85, Gd 6.26;

Found: C, 52.95; H, 5.19; N, 5.48; S, 1.23; F, 12.77; Gd, 6.14.

d) 1- [2- (N-Ethyl-N-perfluorooctylsulfonyl) amino] acetyl-4,10-bis- [10a-D-carbonylmethyl-2,3,4,6-tetra-O-benzylmanopyranose] -7- {pentanoyl -3-aza-4-oxo-5-methyl-5-yl- [1,4,7-tris- (carboxylactomethyl) -1,4,7,10-tetraazacyclododecan-10-yl] -Gd-complex} - 1,4,7,10-tetraazacyclododecane

30.0 g (11.94 mmol) of the title compound of Example 8c are dissolved in a mixture of 300 ml of ethanol and 30 ml of water, 4.0 g of palladium catalyst (10% Pd / C) are added and hydrogenated at room temperature. The catalyst is then filtered off with suction and the filtrate is evaporated to dryness in vacuo. The product was obtained as a colorless solid containing 3.4% water. Yield 21.39 g (100%).

Analysis (calculated on the anhydrous substance):

H, 4.39; N, 7.82; S, 1.79; F, 18.03; Gd, 8.78;

Found: C 36.80, H 4.50, N 7.85, S 1.68, F 17.91, Gd 8.70.

Example 9

a) 6-N- [3,6-Bis (carboxymethyl) octane-1,8-dicarboxylic acid-1-carboxy-8-oyl] -2-N- (1-O-α-D-carboxymethylmanopyranose) -lysine- [ 1- (4-perfluorooctylsulfonyl) -piperazine] -amide

To a solution of 27.5 g (30.0 mmol) of the title compound of Example 1e in a mixture of 300 mL of dimethylformamide and 100 mL of pyridine was added 25.62 g (100.0 mmol) of ethylenediamine-N, N, Ν ', N dianhydride. 'tetraacetic acid, and the mixture was stirred for 5 h at 50 ³ C. After evaporation in vacuo to dryness, the residue is dissolved in 300 ml of water, treated with 20% aqueous NaOH until pH 10 with cDNA and the alkaline product solution was then adjusted with concentrated hydrochloric acid to pH 3 and evaporated to dryness in vacuo. The residue is chromatographed on RP-18 silica gel with water-ethanol-acetonitrile (gradient). 18.22 g (51.0%) of product are obtained as a colorless solid containing 7.9% of science.

Analysis (calculated on the anhydrous substance):

Calculated: C 36.31, H 3.98, N 7.06, F 27.12, S 2.69; Found: C, 36.23; H, 4.07; N, 6.98; F, 27.05; S, 2.62.

b) 6-N- [3,6-Bis (carboxylatomethyl) octane-1,8-dicarboxylic acid-1-carboxylate-8-oyl-Mn-complex, sodium salt of 2-N- (1-OaD) -L-lysine- [1- (4-perfluorooctylsulfonyl; piperazine) amide]

9a the carbonate is adjusted to a filtered solid g (8.397 mmol) of the title compound in 200 ml of water. Manganese (965 mg, 8.397 mmol) was added and stirred at 60 ° C for 3 hours. The solution was pH 7.4 by addition of 5% aqueous sodium hydroxide, and lyophilized. 10.52 g (99.01) of a colorless product containing 7.8% water is obtained.

Analysis (calculated on the anhydrous substance):

Calculated: C 34.16, H 3.50, M 6.64, S 2.53, F 25.52, M 4.34, Na 1.82;

Found: C, 34.06; H, 3.61; N, 6.58; S, 2.47; F, 25.47; M, 4.30; Na, 1.97.

Example 10

(a) 1,2,3,4,6-Penta-O-acetyl-α, β-D-manopyranose

In an analogous manner to that described in the literature [ML Wolfrom and A. Thompson in Methods in Carbohydrate Chemistry (RL Whistler, ML Wolfrom and JN BeMiller, Eds.), Academic Press, New York, Vol. IZ, 53, p. 211-215 (1963) 1, 150 g (832.5 mmol) of α, β-D-manopyranose are reacted with a mixture of 1500 ml of absolute pyridine and 1500 ml of acetic anhydride. After work-up, 315 g (36.7%) of the crude title compound are obtained as a viscous colorless oil. According to the ¹ H-NMR spectrum, the thus obtained product and the β anomers contained 4: 1. It is not necessary to separate the anomers for use in further reaction steps.

analysis:

Calculated: C 49.21, H 5.68;

Found: C, 49.12; H, 5.78.

b) 1-O-α-D- (5-Ethoxycarbonyl) pentyl-2,3,4,6-tetra-O-acetylmanopyranose

In an analogous manner to that described in the literature for the synthesis of arylglycopyranosides [J. Conchie and G.A. Levvy in Methods in Carbohydrate Chemistry (R. L. Wistler, M. L. Wolfrom and J. N. BeMiller, Eds.), Academic Press, New York, Vol. II, 90, p. 345-347 (1963)], a reaction of 156.2 g (400 mmol) of the title compound of Example 10a (as aa and β anomers) with 67 mL (400 mmol) of 6-hydroxyhexanoic acid ethyl ester and 60.8 mL (520 mL) was carried out. mmol) of tin tetrachloride in a total of 600 ml of 1,2-dichloroethane. Purification by silica gel column chromatography (2: 1 hexane-ethyl acetate) gave the title compound as a colorless viscous oil (100.05 g, 51%). According to 1 H-NMR spectroscopy, the product is exclusively pure α-anomer.

analysis:

Calculated: C 52.94, H 6.77;

Found:% C, 52.80;% H, 6.78.

c) 1-O-α-D- (5-Carboxy) pentyl-2,3,4,6-tetra-O-benzylmanopyranose

To a stirred suspension of the title compound of Example 10b (141.0 g, 289 mmol) in dioxane (200 mL) at room temperature under stirring was added a total of 238.5 g (4.26 mol) of finely powdered potassium hydroxide in portions. An additional 200 mL of dioxane was added to the mixture for better mixing. The suspension obtained is finally heated to boiling and a total of 372 ml (3.128 mol) of benzyl bromide is added dropwise at this temperature over 2 hours. After 4 hours at 110 ° C and 12 hours at room temperature, the reaction mixture is poured slowly into 2.5 liters of ice water and the aqueous shock is extracted completely with diethyl ether. After washing the ether phase, drying over sodium sulfate and filtration, the diethyl ether is evaporated in vacuo. Excess benzyl bromide is distilled off quantitatively from the evaporator at 180 ° C (oil bath temperature) under oil pump vacuum. The oily residue thus obtained was purified by chromatography on silica gel (ethyl acetate / hexane 1:10). Yield: 172.2 g (91.0%) of the title compound as a colorless, very viscous oil.

Analysis:

Calculated: C 75.68, H 7.16; Found:% C, 75.79;% H, 7.04.

d) 6-N-Benzyloxycarbonyl-2-N- [1-O-α- (5-carbonyl) pentyl-2,3,4,6-tetra-O-benzylmanopyranose] -L-lysine- [1- (4) -perfluóroktylsuifonyl) -piperazine] amide

100.0 g (134.0 mmol) of the carboxylic acid prepared in Example 10c and 32.4 g (281.4 mmol) of N-hydroxysuccinimide are dissolved in 500 ml of dimethylformamide and added in portions at 0 ° C in total. 1.0 g (281.4 mmol) of N, N'-dicyclohexylcarbodiimide. The mixture was stirred at this temperature for 3 hours. To the thus prepared active ester solution, a cooled solution of 111.3 g (134.0 mmol) of the title compound of Example 1c in 300 ml of dimethylformamide was added dropwise at 0 ° C and the mixture was stirred for 2 hours at 0 ° C and 12 hours at room temperature. The precipitated dicyclohexylurea is filtered off and the filtrate is evaporated to dryness. The residue is chromatographed on silica gel with dichloromethane-ethanol (20: 1; gradient with continuous increase in ethanol). 132.5 g (67.4%) of the title compound are obtained as a colorless, very viscous oil.

analysis:

Calculated: C 54.02, H 4.88, N 3.82, F 22.01, S 2.19; Found: C 53.87, H 4.85, N 4.02, F 22.55, S 2.06.

e) 2-N- [1-O-α-D- (5-carbonyl) pentyl-manopyranose] -L-lysine - [1- (4-perfluorooctylsulfonyl) piperazine] amide

120.0 g (81.77 mmol) of the title compound of Example 10d are dissolved in 800 ml of ethanol, 4.5 g of Pearlman catalyst (20% Pd / C) are added and hydrogenated at room temperature and 1 atm until it absorbs hydrogen (about 8 hours). The catalyst was filtered off with suction, washed well with ethanol (about 200 ml) and the filtrate was evaporated to dryness in vacuo. Yield 78.5 g (98.7%) of a colorless, highly viscous oil.

analysis:

Calculated: C 37.04, H 4.25, N 5.76, F 33.20, S 3.30; Found: C 36.96, H 4.85, N 5.41, F 34.13, S 3.22.

f) 2-N- [1-O-α-D- (5-carbonyl) pentyl-manopyranose] -6-N- [1,4,7-tris (carboxylatomethyl) -10- (3-aza-4-oxo) 5-Methyl-5-yl-pentanoyl) -1,4,7,10-tetraazacyclododecane] -L-lysine- [1- (4-perfluoro-oylsulfonyl) -piperazine] -amide, gadolinium complex

99.8 g (158.4 mmol; 2.2 mol equiv., Based on the amine component of Example 10e) gadolinium complex of 10- (4-carboxy-1-methyl-2-oxo-3-azabutyl) -1,4, 7,10-tetraazacyclododecane-1,4,7-triacetic acid (described in patent application

DE 197 28 954 Cl in Example 31h) and 6.7 g of anhydrous lithium chloride (158.4 mmol) were dissolved in 800 ml of absolute dimethylsulfoxide at 40 ° C with stirring. A total of 18.2 g (158.4 mmol) of N-hydroxysuccinimide and 70.0 g (71.96 mmol) of the title compound of Example 10e dissolved in 250 ml of absolute dimethylsulfoxide are then added at this temperature. After cooling to room temperature, 32.7 g (158.4 mmol) of N, N'-dicyclohexylcarbodiimide was added to the reaction solution, and the mixture was stirred at room temperature for 12 hours. the precipitate is filtered off with suction, dried, admixed with water, filtered, insoluble dicyclohexylurea and the filtrate is desalted and freed from low molecular weight components on an Amicon® YM-3 ultrafiltration membrane (cut off: 3000 Da). 0 g (81.6%) of a colorless lyophilizate containing 9.53% water (Kari-Fischer).

Analysis (calculated on the anhydrous substance):

Calculated: C 37.15, H 4.39, N 7.96, F 20.38, S 2.02, 9.92%

Gd;

Found: C 36.92, H 4.50, N 7.68, F 19.77, S 1.91, Gd 10.08%.

Example 11

a) 2-N- [1-OaD- (5-carbonyl) pentyl-mannopranose] -6-N- (2- [4- (3-oxapropionyl) phenyl] -2- [1,4,7-tris] (carboxylateomethyl) -1,4,7,10-tetraacyclododecan-10-yl] acetic acid) -L-lysine- [1- (4-perfluorooctylsulfonyl) piperazine] amide, gadclinium complex, sodium salt

To a stirred suspension of 5.0 g (9.06 mmol) of the title compound of Example 3e in 15 mL of absolute dimethylsulfoxide at 70 ° C was added 0.68 g (15.9 mmol) of lithium chloride. After stirring at 70 ° C for thirty minutes, a total of 1.83 g (15.9 mmol) of N-hydroxysuccinimide was added portionwise to the resulting clear reaction solution, and the reaction mixture was maintained at this temperature for 1 hour. After cooling to 0 ° C, dicyclohexylcarbodiimide (4.52 g, 23.85 mmol) was added and the reaction solution was stirred at 0 ° C for an additional hour and then at 22 ° C for 12 hours. A solution of 4.0 g (4.12 mmol) of the title compound of Example 10e in 15 mL of absolute dimethylsulfoxide was added dropwise at 22 ° C to the thus obtained N-hydroxysuccinimide ester ester solution of Example 3e and stirred for an additional 12 hours at room temperature. The reaction solution was then added dropwise at 22 ° C to 900 ml of acetone, whereupon the title compound precipitated as a colorless precipitate. The precipitate is filtered off with suction, dissolved in 200 ml of distilled water and adjusted to pH 7.2 by addition of 1M sodium hydroxide. The aqueous product solution thus obtained is subjected to a three-fold ultrafiltration through a YM3 ultrafiltration membrane (Amicon®, cut off: 3000 Da) to achieve desalting and to remove the low molecular weight components. The retentate is then lyophilized. Yield 6.33 g (92.4% based on the charged amine component) of the product as a colorless lyophilisate containing 7.38% water.

Analysis (calculated on the anhydrous substance):

Calculated: C 38.48, H 4.13, N 6.65, F 19.16, S 1.90, Gd 9.33, Na 1.36;

Found: C 39.52, K 4.12, N 6.67, F 19.70, S 1.89, Gd 9.30, Na 1.41.

Example 12

a) 3,5-bis (benzyloxycarbonylamino) benzoic acid N- (3-oxa-1H, 1H, 2H, 2H, 4H, 4H, 5H, 5H-perfluorotridyl) amide

20 g (47.5 mmol) of 3,5-bis (benzyloxycarbonylamino) benzoic acid (prepared according to: Skulnick, Harvey I .; Johnson, Paul D .; Aristoff, Paul) are dissolved in a mixture of 125 ml of dry tetrahydrofuran and 125 ml of dry dioxane. A. Morris, Jeanette K., Lovasz, Kristine D. et al .: J. Med. Chem. 40; 7; 1997; 1149-1164) and 4.78 g (47.5 mmol) of triethylamine. After cooling to -15 ° C, a solution of 6.56 g (48 mmol) of isobutyl chloroformate in 30 ml of dry tetrahydrofuran is slowly added dropwise with stirring, keeping the internal temperature below -10 ° C. After 15 minutes at -15 ° C, a solution of 58.6 g (47.5 mmol) of 1-amino-1H, 1H, 2H, 2H, 4H, 4H, 5H, 5H-3-oxaperfluorotridecane, and 4.78 g (47.5 mmol) of triethylamine in 100 ml of dry tetrahydrofuran. After 1 hour at -15 ° C and 2 hours at room temperature, the reaction solution is evaporated to dryness in vacuo. The residue is taken up in 300 ml of ethyl acetate and extracted twice with 200 ml of saturated sodium bicarbonate solution and once with 300 ml of water. The organic phase is dried over sodium sulphate, filtered and evaporated in vacuo. The oily residue was purified on silica gel with dichloromethane-hexane-2-propanol (10: 5: 1) to give 36.2 g (82.5%) of the title compound as a colorless oil.

analysis:

H, 3.27; N, 4.55; F, 34.97.

Found: C 47.21, H 3.31, N 4.61, F. 34.48.

b) 3,5-diaminobenzoic acid N- (3-Oxa-1 ', 1H, 2H, 2H, 4H, 4H, 5H, 5H-perfluorotridecyl) amide

30.0 g (32.4 mmol) of the amide prepared in Example 12a are dissolved in 300 ml of ethanol, 1.2 g of Pearlman catalyst (20% Pd / C) are added and hydrogenated at room temperature and 1 atm, when hydrogen is absorbed. The catalyst is then filtered off with suction, washed thoroughly with ethanol (about 300 ml) and the filtrate is evaporated to dryness in vacuo. Yield 20.12 g (94.8%) of a yellowish very viscous oil.

analysis:

Calculated: C 36.66, H 2.77, N 6.41, F 49.28;

Found: C 36.07, H 2.87, N 6.23, F 49.43.

c) 3-N - [(1-Oa-D-carbonylmethyl-2,3,4,6-) - (3-Oxa-1 H, 1 H, 2 H, 2 H, 4 H, 4 H, 5 H, 5 H -fluorotridecyl) amide tetra-O-benzyl-r.anopyranóza) -5-aminobenzoic acid

10.95 g (18.30 mmol) of 1-carboxymethyloxy-2,3,4,6-tetra-O-benzyl-α-D-manopyranoside (prepared as described in DE 197 28 954 C1) are dissolved in 150 ml of dimethylformamide to this solution was added a total of 2.09 g (18.3 mmol) of N-hydroxysuccinimide. The mixture was cooled to 0 ° C and dicyclohexylcarbodiimide (3.78 g, 18.3 mmol) was added. The mixture was stirred at 0 ° C for 1 hour and then at room temperature for 4 hours. It is then cooled to 0 [deg.] C. and a solution of 24.0 g (36.6 mmol; 2 mol equivalents, based on the carboxylic acid charged) of the diamino compound described in Example 12b in 350 ml of dimethylformamide is slowly added dropwise over 3 hours. The mixture was stirred for 1 hour at 0 ° C and then overnight at room temperature. After evaporation in vacuo to dryness, the residue is taken up in 300 ml of ethyl acetate, filtered from the precipitated urea and the filtrate is washed twice with 100 ml of 5% aqueous soda solution. The organic phase is dried over magnesium sulphate and evaporated to dryness in vacuo. The residue was chromatographed on silica gel with n-hexane-isopropanol (13: 1). It will be obtained

16.8 g (74.3% based on carboxylic acid) of the title compound as a colorless oil. Increasing the polarity of the eluent to n-hexane-isopropanol (5: 1) gave a total of 10.15 g of unreacted diamino compound 12b in further chromatographic fractions, which could be reused in the above reaction.

analysis:

Calculated: C 54.42, H 4.40, N 3.40, F 26.13;

Found: C, 54.32; H, 4.49; N, 3.48; F., 25.94.

d) 3-N - [(1-O-α-D-carbonylmethyl-mannopyranose)] - 5-aminobenzoic acid N- (3-oxa-1H, 1H, 2H, 2H, 4H, 4H, 5H, 5H-perfluorotridecyl) amide

Analogously to the synthesis of the title compound of Example 12b, hydrogenolysis of 12.0 g (9.70 mmol) of the title compound of Example 12c in the presence of 0.5 g of Pearlman's catalyst (20% Pd / C) in ethanol-water (9). : 1) after work-up, gave 8.08 g (96.7%) of the title compound as a yellowish viscous oil.

analysis:

Calculated: C 37.64, H 3.28, N 4.88, F 37.49;

Found: C, 37.32; H, 3.17; N, 4.97; F. 37.55.

e) 3-N- [(1-Oa-D-carbonylmethyl-manopyranose) -5-N- {2-N- [3-oxa-1H, 1H, 2H, 2H, 4H, 4H, 5H, 5H-perfluorotridecyl] amide] - [4- (3-oxapropionyl) phenyl] -2- [1,4,7-tris (carboxylatomethyl) -1,4,7,10-zetraazacyclododecan-10-yl] acetic acid} benzoic acid, gadolinium complex, sodium salt

13.6 g (19.2 mmol); 2.2 mol equivalents relative to the amine component of Example 12d) of the gadolinium complex described in Example 3e and 0.81 g (19.2 mmol) of anhydrous lithium chloride are dissolved in 100 ml of anhydrous dimethylsulfoxide and stirred at 40 ° C. a total of 2.2 g (19.2 mmol) of N-hydroxysuccinimide and 7.5 g (8.7 mmol) of the title compound of Example 12d in 50 mL of absolute dimethylsulfoxide are added at this temperature. After cooling to room temperature, 3.96 g (19.2 mmol) of N, N'-dicyclohexylcarbodiimide was added to the reaction solution, and the mixture was stirred at room temperature for 12 hours. Sufficient acetone is then added to the resulting suspension to completely precipitate the title compound, the precipitate is filtered off with suction, dried, stirred with water, filtered from insoluble dicyclohexylurea and the filtrate is desalted and freed from the low molecular weight components using an Amicon® YM-3 ultrafiltration membrane. (cut off 3000 Da). The retentate was lyophilized to give 11.51 g (84.5%) of the product as a colorless lyophilisate containing 6.77% water (Kari-Fischer).

Analysis (calculated on the anhydrous substance):

H, 3.94; N, 6.29; F, 20.71; G, 10.08; Na, 1.47;

Found: C, 39.98, H, 4.00, N, 6.31, F, 20.73, Gd, 10.11, Na, 1.42.

Example 13

a) 3,5-Bis (benzyloxycarbonylamino) -1- {N - [1- (4-perfluorooctylsulfonyl) piperazine] [benzamide

10 g (23.75 mmol) of 3,5-bis (benzyloxycarbonylamino) benzoic acid (prepared according to: Skulnick, Harvey I .; Johnson, Paul D .; Aristoff, Paul) are dissolved in a mixture of 60 ml of dry tetrahydrofuran and 70 ml of dry dioxane. A. Morris, Jeanetre K., Lovasz, Kristin D., et al., J. Med. Chem. 40; 7; 1997; 1149-1164) and 2.39 g (23.75 mmol) of triethylamine. After cooling to -15 ° C, a solution of 3.28 g (24 mmol) of isocutyl chloroformate in 20 ml of dry tetrahydrofuran is slowly added dropwise with stirring so that the internal temperature does not exceed -10 ° C. After 15 minutes at -15 ° C, a solution of 23.0 g (23.75 mmol) of perfluorooctylsulfonylpiperazine and 2.39 g (23.75 mmol) of triethylamine in 50 ml of dry tetrahydrofuran is added dropwise at -20 ° C. After 1 hour at -15 ° C and 2 hours at room temperature, the reaction solution is evaporated to dryness in vacuo. The residue is taken up in 200 ml of ethyl acetate and washed twice with 100 ml of saturated sodium bicarbonate solution and once with 300 ml of water. The organic phase is dried over sodium sulphate, filtered and the ethyl acetate is evaporated in vacuo. The oily residue was purified by chromatography on silica gel with dichloromethane-hexane-2-propanol (15: 5: 1) to give 13.35 g (79.6%) of the title compound as a colorless oil.

analysis:

Calculated: 43.31% C, 2.80% H, 5.77% N, 33.27% 1 ', 3.30% S; Found: C 43.21, H 2.75, N 5.61, F 33.38, S 3.22.

b) 3,5-Diamino-1- {N- [1- (4-perfluorooctylsulfonyl) piperazine) oenzamide

The amide of Example 13a (9.70 g, 10.0 mmol) was dissolved in ethanol (100 mL), Pearlman catalyst (20% Pd / C) (0.4 g) was added and hydrogenated at room temperature and 1 atm until it absorbs hydrogen. The catalyst is filtered off with suction, washed thoroughly with ethanol (about 150 ml) and the filtrate is evaporated to dryness in vacuo. Yield 6.9 g (98.2%) of a yellowish very viscous oil.

analysis:

Calculated: C 32.49, H 2.15, N 7.98, F 45.98, S 4.56; Found: C 32.56, H 2.17, N 8.09, F 45.63, S 4.61.

c) 5-Amino-3-Ν- [1-O-α-D-carbonylmethyl-2,3,4,6-tetra-O-benzyl-manccyranose) benzoic acid N- [1- (4-perfluorooctylsulfonyl) piperazine] amide

5.48 g (9.15 mmol) of 1-carboxymethyloxy-2,3,4,6-tetra-O-benzyl-α-D-manopyranoside (prepared as described in the file)

DE 197 28 954 C1) is dissolved in 100 ml of DMF and to this solution is added a total of 1.04 g (9.15 mmol) of N-hydroxysuccinimide. The mixture was cooled to 0 ° C and dicyclohexylcarbodiimide (1.89 g, 9.15 mmol) was added. The mixture was stirred at 0 ° C for 1 hour and then at room temperature for 4 hours. It is then recooled to 0 ° C and a solution of 12.85 g (18.30 mmol; 2 mol equivalents, based on the carboxylic acid) of the diamino compound described in Example 13b in 250 ml of dimethylformamide is slowly added dropwise over 3 hours. The mixture was stirred for 1 hour at 0 ° C and then overnight at room temperature. After evaporation in vacuo to dryness, the residue is taken up in 100 ml of ethyl acetate, filtered from the precipitated urea and the filtrate is washed twice with 100 ml of 5% aqueous soda solution. The organic phase is dried over magnesium sulphate and evaporated to dryness in vacuo. The residue was chromatographed on silica gel with n-hexane-isopropanol (13: 1). Yield: 8.14 g (69.4% based on carboxylic acid) of the title compound as a colorless oil. Increasing the polarity of the solvent elution mixture to n-hexane-isopropanol (6: 1) gave a total of 4.36 g of unreacted diamino compound 13b, which could be reused in the reaction described above, in further chromatographic fractions.

analysis:

Calculated: C 51.49, H 4.01, N 4.37, F 25.17, S 2.50; Found:% C, 51.60;% H, 4.19;% N, 4.28.% F, 2.44%.

d) 5-Amino-3-Ν- [1-O-α-D-carbonylmethyl-mannopyranose) benzoic acid N- [1- (4-perfluorooctylsulfonyl) piperazine] amide

Analogously to the title compound of Example 13b, hydrogenolysis of 6.4 g (5.0 mmol) of the title compound of Example 13c in the presence of 0.3 g of Pearlman's catalyst (20% Pd / C) in ethanol-water (8: 1) gave after working up 4.43 g (96.2%) of the above title compound as a yellowish viscous oil.

Analysis:

Calculated: C 35.15%, H 2.95%, N 6.07%, F 35.01%, S 3.48%; Found: C 35.32, H 3.02, N 5.89, F 35.05, S 3.58.

e) N- [1- (4-Perfluorooctylsulfonyl) piperazine] -amide 3-N- [1---α-D-carbonylmethyl-manopyranose) -5-N- [1,4,7-tris (carboxylatomethyl) - 10- (3-aza-4-oxo-5-methyl-5-yl-pentanoyl) -1,4,7,10-tetraazacyclododecane] benzoic acid

5.54 g (8.8 mmol; 2.2 mol equivalents, based on the amine component used in Example 13d) of the gadolinium complex of 10- (4-carboxy-1-methyl-2-oxo-3-azabutyl) -1,4 7,10-tetraazacyclododecane-1,4,7-triacetic acid (described in patent application

DE 197 28 954 Cl in Example 31h) and 0.37 g (8.8 mmol) of anhydrous lithium chloride are dissolved in 60 ml of absolute dimethylsulfoxide with stirring at 40 ° C and a total of 1.01 g (8, 8 mmol) of N-hydroxysuccinimide and 3.7 g (4.0 mmol) of the title compound of Example 13d in 40 ml of absolute dimethylsulfoxide. After cooling to room temperature, 1.82 g (8.8 mmol) of N, N'-dicyclohexylcarbociimide was added to the reaction solution, and the mixture was stirred at room temperature for 12 hours. Sufficient acetone is then added to the resulting slurry to completely precipitate the title compound, the precipitate is filtered off with suction, dried; it is mixed with water, filtered from insoluble dicyclchexylurea and the filtrate is desalted and freed from low molecular weight components using an Amicon® YM-3 ultrafiltration membrane (cut off 3000 Da). The retentate was lyophilized to give 5.36 g (87.4%) of the product as a colorless lyophilisate containing 6.77% water (Karl-Fischer).

Analysis (calculated on the anhydrous substance):

H, 3.61; N, 8.22; F, 21.05; G, 10.25.

2.09% S;

Found: C 35.87, H 3.70, N 8.22, F 20.91, Gd 10.18, 2.16%

WITH.

Example 14

a) 1,4,7-Triazaheptane-1,7-bis (2-N-trifluoroacetyl-6-N-benzyloxycarbonyl-L-lysine) diamide

100 g (107.9 mmol) of the carboxylic acid prepared in Example 1a and 26.1 g (226.59 mmol) of N-hydroxysuccinimide are dissolved in 500 ml of dimethylformamide. A total of 46.7 g (226.59 mmol) of N, N'-dicyclohexylcarbodiimide was added portionwise at 0 ° C and the mixture was stirred at this temperature for 3 hours. To the thus prepared active ester solution was added dropwise a solution of 5.57 g (53.95 mmol) of diethylenetriamine in 60 ml of dimethylformamide precooled to 0 ° C and the reaction mixture was stirred at 0 ° C for 2 hours and then at room temperature for 12 hours. The precipitated dicyclohexylurea is filtered off and the filtrate is evaporated to dryness. The residue is chromatographed on silica gel with dichloromethane-ethanol (15: 1) using a solvent gradient with a continuous increase in ethanol content. Yield: 26.0 g (58.8% based on amine component) as a colorless, very viscous oil.

analysis:

Calculated: C 52.74, H 5.78, N 11.96, F 13.90;

Found:% C, 52.66;% H, 5.89;% N, 11.88.

b) 1,7-Bis (2-N-trifluoroacetyl-6-N-benzyloxycarbonyl-L-lysine) -diamide 4- [2- (N-ethyl-N-perfluorooctylsulfonyl) amino] acetyl-1,4,7 -triazaheptán

To a solution of 20 g (24.4 mmol) of the diamide prepared in Example 14a in a mixture of 150 ml of tetrahydrofuran and 15 ml of chloroform at 0 ° C under a nitrogen atmosphere is added a solution of 16.13 g (27.0 mmol) of 2- [ Of N-ethyl-N-perfluorooctylsulfonyl] aminoacetic acid (prepared according to DE 196 03 033) in 50 ml of tetrahydrofuran. A total of 18.0 g (36.6 mmol) of EEDQ [2-ethoxy-1-ethoxycarbonyl-1,2-dihydroquinoline] was then added portionwise at 0 ° C and the mixture was stirred overnight at room temperature. After concentration in vacuo, the residual oil under 65 was chromatographed on silica gel with n-hexane-isopropanol (15: 1). 24.74 g (72.4% based on the charged secondary amine) of the title compound are obtained as a colorless oil.

analysis:

Calculated: C 42.01, H 3.96, F 31.19, N 8.00, S 2.29; Found: C 41.92, H 4.07, F 31.22, N 7.87, S 2.34.

c) 1,7-Bis (6-N-benzyloxycarbonyl-L-lysine) diamide 4- [2- (N-ethyl-N-perfluorooctylsulfonyl) amino] acetyl-1,4,7-triazaheptane

22.0 g (15.7 mmol) of the compound prepared in Example 14b are dissolved in 100 ml of ethanol. To this solution was added ammonia gas for 40 minutes at 0 ° C. The mixture was stirred at 0 ° C for 4 hours and then at room temperature for 3 hours. It is then evaporated to dryness in vacuo (bath temperature 40 ° C). The oily residue was purified by chromatography on silica gel with dichloromethane-hexane-2-propanol (20: 10: 1). Yield: 12.92 g (98.4% of theory) of the title compound as a colorless, very viscous oil.

analysis:

Calculated: C 44.22, H 4.64, N 9.38, S 2.68, F 27.03; Found: C 44.31, K 4.72, N 9.30, S 2.74, F. 26.99.

d) 1,7-Bis [6-N-benzyloxycarbonyl-2-N- (1-Oa-D-carbonylmethyl-2,3,4,6-tetra-O-benzyl-manopyranose) -L-lysine] diamide-4- [2- (N-ety1-N-perfluorooctylsulfonyl) -amino] -acetyl-1,4,7-triazaheptane

To a solution of 5.47 g (9.15 mmol) of 1-carboxymethyloxy-2,3,4-tetra-O-benzyl-α-D-manopyranoside (prepared according to DE 197 28 954 C1) in 80 ml of dimethylformamide is added a total of 1 0.05 g (9.15 mmol) of N-hydroxysuccinimide. The mixture is cooled to 0 ° C, 1.9 g (9.15 mmol) of dicyclohexylcarbodiimide is added and stirred for one hour at 0 ° C and then for 4 hours at room temperature. The mixture was cooled to 0 ° C and a solution of 7.65 g (9.15 mmol) of the amino compound described in Example was added dropwise over 3 hours.

14c, in 50 ml of DMF. The mixture was stirred for an additional hour at 0 ° C and then overnight at room temperature. After evaporation to dryness in vacuo, the residue is taken up in 100 ml of ethyl acetate, the precipitated urea is filtered off and the filtrate is washed twice with 50 ml of 5% aqueous soda solution. The organic phase is dried over magnesium sulphate and evaporated to dryness in vacuo. The residue was chromatographed on silica gel with n-hexane-isoprcpanol (20: 1). Yield: 17.01 g (78.9% based on the charged acid) of the title compound as a colorless oil.

analysis:

Calculated: C 59.13, H 5.43, N 4.76, F 13.71, S 1.36; Found: C 59.22, H 5.39, N 4.85, F 13.70, S 1.40.

(e) 1,7-Bis [2-N- (1-O-α-D-carbonylmethyl-manopyranose) -L-lysine] diamide-4- [2- (N-ethyl-N-perfluorooctylsulfonyl) amino] acetyl-1, 4,7-triazaheptan

To a solution of 15.0 g (6.36 mmol) of the amide prepared in Example 14d in 150 mL of ethanol was added 0.5 g of Pearlman's catalyst (20% Pd / C) and hydrogenated at room temperature and at 1 atm pressure as long as no more hydrogen is consumed. The catalyst is filtered off with suction, washed thoroughly with ethanol (about 100 ml) and the filtrate is concentrated to dryness in vacuo. 8.54 g (97.2%) of the title compound are obtained as a very viscous, yellowish oil.

analysis:

Calculated: C 39.13, H 5.04, N 8.11, F 23.38, S 2.32; Found:% C, 39.07;% H, 4.98;% N, 8.18;% F, 2.40%.

f) 1,7-Bis [2-N- (1-Oa-D-carbonyldimethyl-mannopyranose) -6-N- [1,4,7-tris (carboxylatomethyl) -10- (3-aza-4-oxo-5) methyl-5-yl-pentanoyl) -1,4,7,10-tetraazacyclododecane] -L-lysine] -diamide-4- [2- (N-ethyl-N-perfluorooctylsulfonyl) -amino] -acetyl-1,4, 7-triazaheptane, digadolinium complex

To a stirred suspension of 5.7 g (9.06 mmol) of gadolinium complex of 10- (4-carboxy-1-methyl-2-oxo-3-a-butyutyl) -1,4,7,10-tetraazacyclododecane-1,4 Of 7-triacetic acid (described in patent application DE 197 28 954 C1 in Example 31h) in 75 ml of absolute dimethylsulfoxide at 70 ° C is added 0.68 g (15.9 mmol) of lithium chloride. After stirring for 30 minutes at 70 ° C, a total of 1.83 g (15.9 mmol) of N-hydroxysuccinimide was added portionwise to the resulting clear reaction solution, and the reaction mixture was maintained at this temperature for 1 hour. After cooling to 0 ° C, dicyclohexylcarbodiimide (4.52 g, 23.85 mmol) was added and the reaction solution was stirred at 0 ° C for an additional hour and then at 22 ° C for 12 hours. To the thus obtained solution of 10- (4-carboxy-1-methyl-2-oxo-3-azabutyl) -1,4,7,10-tetraazacyclododecane-1,4,7-triacetic acid N-hydroxysuccinimide ester at 22- A solution of 2.84 g (2.06 mmol) of the title compound of Example 14e in 15 mL of absolute dimethylsulfoxide is added dropwise and the mixture is stirred for another 12 hours at room temperature. The reaction solution was then added dropwise at 22 ° C to 500 ml of acetone, whereupon the title compound precipitated as a colorless precipitate. The precipitate is filtered off with suction, dissolved in 200 ml of distilled water and subjected to three-fold ultrafiltration through a YM-3 ultrafiltration membrane (Amicon®, cut off: 3000 Da) to achieve desalination and to remove low molecular weight components. The retentate obtained is then lyophilized. 4.80 g (89.6% based on the charged amine component) of the product are thus obtained in the form of a colorless lyophilisate which contains 8.98% of water.

Analysis (calculated on the anhydrous substance):

Calculated: C 38.28, H 4.84, N 9.68, E 12.40, S 1.23, Gd 12.07;

Found: C, 38.20; H, 4.91; N, 9.77; E, 12.45; S, 1.19; Gd, 12.10.

Example 15

a) 1,7-bis (benzyloxycarbonyl) -4- (3-oxapentane-1,5-dicarboxylic acid-1-yl-5- [1- (4-perfluorooctylsulfonyl) piperazine] amide) -1,4,7,10 -tetraazacyklododekán

To a solution of 1,7-bis (benzyloxycarbonyl) -1,4,7,10-tetraazacyclododecane (10.75 g, 24.4 mmol) in a mixture of 150 ml of tetrahydrofuran and 15 ml of chloroform at 0 ° C under nitrogen is added a solution of 16, A total of 18.0 g of EEDQ (2-ethoxy-1-ethoxycarbonyl-1,2-dihydroquinoline) was added portionwise at 0 ° C, 56 g (24.4 mmol) of the title compound of Example 15e in 150 ml of zetraPot was added and stirred through overnight at room temperature. After evaporation in vacuo, the residual oil is chromatographed on silica gel with n-hexane-isopropanol (12: 1). 17.22 g (64.3% of the charged hydrofuran (36.6 mmol) of the secondary amine) of the monoamide and g (8.8%) of the diamide are obtained. The resulting compound is a colorless oil.

analysis:

H, 3.92; F, 29.18; N, 7.59; S, 2.60. Found:% C, 43.52;% H, 4.07;% F, 29.24;% N, 7.67.

b) 1,7-Bis (benzyloxycarbonyl) -4- (3-oxapentane-1,5-dicarboxylic acid-1-oyl-5- [1- (4-perfluorooctylsulfonyl) piperazine] amino) -10- [1-0 -αD- (5-carbonyl) pentyl-2,3,4,6-tetra-O-benzylmanopyranose] -1,4,7,10-tetraazacyclododecane

10.0 g (13.4 mmol) of the carboxylic acid prepared in Example 10c and 3.24 (28.1 mmol) of N-hydroxysuccinimide are dissolved in 100 ml of dimethylformamide. A total of 5.8 g (28.1 mmol) of N, N'-dicyclohexylcarbodiimide was added portionwise at 0 ° C and the mixture was stirred at this temperature for 3 hours. To the thus prepared active ester solution was added dropwise a solution of 14.83 g (13.4 mmol) of the title compound of Example 15a in 100 ml of dimethylformamide, precooled to 0 ° C, and the reaction mixture was stirred for 2 hours at 0 ^{° C} and then for 12 hours at room temperature. The precipitated dicyclchexylurea is filtered off and the filtrate is evaporated to dryness. The residue is chromatographed on silica gel with dichloromethane-ethyl acetate (20: 1), using a gradient with a continuous increase in the ethyl acetate content. 18.3 g (78.2%) of the title compound are obtained as a colorless, very viscous oil.

analysis:

H, 5.03; N, 4.82; F, 18.52; S, 1.84; Found: C, 54.87, H, 4.85, N, 4.92, F, 18.55, S, 1.86.

c) 1- {3-Oxapentane-1,5-dicarboxylic acid-1-oyl-5- [1- (4-perfluorooctylsulfonyl) piperazine] amide} -7- [1-CaD- (5-carbonyl) pentyl-manopyranose -1,4,7,10-tetraazacyclododecanoate

To a solution of 17.0 g (9.65 mmol) of the compound prepared in Example 14b in 150 mL of ethanol was added 1.0 g of Pearlman's catalyst (20% Pd / C) and hydrogenated at room temperature under 1 atm pressure. as long as hydrogen is no longer consumed. The catalyst is filtered off with suction, washed thoroughly with ethanol (2 x 75 ml) and the filtrate is concentrated to dryness in vacuo. Yield: 10.76 g (99.0% of theory) of the title compound as a very viscous colorless oil.

analysis:

Calculated: C 38.78, H 4.61, N 7.54, F 28.97, S 2.88; Found: C 38.86%, H 4.65%, N 7.41%, F 29.02%, S 2.92%.

d) 1,7-Bis [1,4,7-tris (carboxylatomethyl) -1,4,7,10-tetraazacyclododecane-Gd complex-10- (pentanoyl-3-aza-4-oxo-5-methyl- 5-yl) -4- [2-N-ethyl-N-perfluorooctylsulfonyl) amino] acetyl-2-oxa-acetyl] -10- [10aD-6-carbonylpentyl-manopyranose] -1,4,7,10-tetraazacyclododecane

24.86 g (39.46 mmol; 4.4 mol equivalents based on the amine component 15c) of the gadclinic complex of 10- (4-carboxy-1-methyl-2-oxo-3-azabutyl) -1,4,7, 10-tetraazacyclododecane-1,4,7-triacetic acid (described in patent application

DE 197 28 954 Cl in Example 31h) and 1.67 g (39.46 mmol) of lithium chloride are dissolved with stirring at 40 ° C in 200 ml of absolute dimethylsulfoxide and a total of 4.53 g (39.46) is added at this temperature. mmol) of N-hydroxysuccinimide and 10.0 g (8.97 mmol) of the title compound of Example 14c in 100 mL of absolute dimethylsulfoxide. After cooling to room temperature, 8.14 g (39.46 mmol) of dicyclohexylcarbodiimide was added to the reaction solution and stirred for 12 hours at room temperature. Sufficient acetone is added to the suspension thus obtained to completely precipitate the title compound, the precipitate is filtered off with suction, dried, mixed with distilled water, the insoluble dicyclohexylurea is suctioned off and the filtrate is subjected to ultrafiltration through an Amiccn® YM-3 ultrafiltration membrane ( cut off: 3000 Da) to obtain desalination and remove low molecular weight components. The retentate is then lyophilized. 16.37 g (79.3%) of product are obtained in the form of a colorless lyophilisate which contains 7.65% of water (Karl -ischer).

Analysis (calculated on the anhydrous substance):

Calculated: 38.01% C, 4.61% n, 9.58% N, 13.31% F, 1.37% S, 13.45% Gd;

Found: C 37.92, H 4.55, N 9.58, F 13.77, S 1.31, Gd 13.48.

e) 3-Oxapentane-1,5-dicarboxylic acid mono- [1- (4-perfluorooctylsulfonyl) piperazine] amide

To a solution of 25 g (44.0 mmol) of 1-perfluorooctylsulfonylpiperazine in 150 ml of tetrahydrofuran at room temperature was added a total of 5.1 g (44.0 mmol) of diglycolic anhydride and the reaction solution was refluxed for 12 hours. After cooling to room temperature, it is evaporated to dryness and the oily residue is purified by chromatography on silica gel using dichloromethane-2-propanol (16: 1). Yield: 27.94 g (92.8%) of the title compound as a colorless viscous oil.

analysis:

Calculated: C 58.52, H 4.27, N 1.98, S 2.26, F 22.80;

Found: C 58.42, H 4.41, N 1.80, S 2.28, F. 23.02%.

Example 16

a) 1,7-Bis (benzyloxycarbonyl) -4- (3-oxapenran-1,5-dicarboxylic acid-1-oyl-5- [1- (4-perfluorooctylsulfonyl) piperazine amide) -10- [Ι-Ο-] β-D- (6-carbonyl) pentyl-2,3,4,6-tetra-O-benzylglucopyrano-1] -1,4,7,10-tetraazacyclododecane

68.5 g (91.79 mmol) of 1-carboxymethyloxy-2,3,4-tetra-O-benzyl-α-D-manopyranoside (prepared according to DE 19728954 C1) are dissolved in 750 ml of dry tetrahydrofuran and the solution Add 9.25 g (91.79 mmol) of triethylamine. The reaction solution is cooled to -15 ° C to -20 ° C and a solution of 12.64 g (92.5 mmol ·) of isobutyl chloroformate in 150 ml of dry tetranydrofuran is slowly added dropwise at this temperature to the internal temperature with stirring. does not exceed -10 ° C. After the addition, the mixture was kept at -15 ° C for 15 minutes and then a solution of 101.6 g (91.79 mmol) of the title compound of Example 15a and 9.25 g (91.79 mmol) of triethylamine was slowly added dropwise at -20 ° C. in 500 ml dry tetrahydrofuran. After 1 hour at -15 ° C and 2 hours at room temperature, the reaction solution is evaporated to dryness in vacuo. The residue is taken up in 450 ml of ethyl acetate and the solution is washed with 2 x 300 ml of saturated sodium bicarbonate solution and then once with 400 ml of water. After drying over sodium sulfate, it is filtered and the ethyl acetate is evaporated under vacuum. The oily residue was purified on silica gel with dichloromethane-hexane-2-propanol (10: 20: 1). 130.6 g (81.6%) of the title compound are obtained as a colorless, very viscous oil.

analysis:

H, 5.03; N, 4.82; F, 18.52; S, 1.84; Found: C 55.20, H 5.09, N 4.91, F 18.48, S 1.80.

b) 1- {3-Oxapentane-1,5-dicarboxylic acid-1-oyl-5- [1- (4-per-fluorooctylsulfonyl) piperazine] amide] -7- [1-O-α- (5-carbonyl)] pentyl-manopyranose] -1,4,7,10-tetraazacyclododecane

To a solution of 110.0 g (63.08 mmol) of the compound prepared in Example 16a in 1000 mL of ethanol was added 5.0 g of Pearlman catalyst (20% Pd / C) and hydrogenated to quantitatively consume hydrogen. The catalyst is filtered off with suction, washed with ethanol and the filtrate is evaporated to dryness in vacuo. Yield 92.61 g (99.5%) of the title compound as a viscous colorless oil.

analysis:

Calculated: C 52.10, H 5.12, N 5.70, F 21.89, S 2.17; Found: C, 52.20; H, 5.09; N, 5.71; F, 21.87; S, 2.20.

c) 1,7-Bis [1,4,7-tris (carboxylatomethyl) -10- (3-aza-4-oxo-5-methyl-5-ylpentanoyl) -4- (3-oxapentane- -1,5,7,10-tetraazacyclododecane, 1,5-dicarboxylic acid-1-oyl-5- [1- (4-perfluorooctylsulfonyl) piperazine] amide} -10- [1-OaD-5-carbonylpentyl-manopyranose], digadolinium complex

55.4 g (88.0 mmol; 4.4 mol equivalents based on the charged diamine component of Example 13d) of the gadolinium complex of 10- (4-carboxy-1-methyl-2-oxo-3-azabutyl) -1,4 7,10-tetraazacyclododecane-1,4,7-triacetic acid (described in patent application DE 197 28 954 Cl in Example 31h) and 3.7 g (88.0 mmol) of anhydrous lithium chloride are dissolved with stirring at 40 ° C. in 500 ml of absolute dimethylsulfoxide and at this temperature a total of 10.1 g (88.0 mmol) of N-hydroxysuccinimide and 29.5 g (20.0 mmol) of the title compound of Example 16b in 200 ml of absolute dimethylsulfoxide are added. After cooling to room temperature, 18.2 g (88.0 mmol) of N, N'-dicyclohexylcarbodiimide are added to the solution and stirred for 12 hours at room temperature. Sufficient acetone is added to the resulting suspension to completely precipitate the title compound, the precipitate is filtered off with suction, dried, dissolved in water, the insoluble dicyclohexylurea is filtered off with suction and the filtrate is filtered through an Amicon® YM-3 ultrafiltration membrane (cut off: 3000 Da). to desalinate and remove the low molecular weight components. The retentate is then lyophilized. 35.96 g (76.9%) of the product are obtained in the form of a colorless lyophilisate which contains 5.98% of water (Kari-Fischer).

Analysis (calculated on the anhydrous substance):

Calculated: C 38.01, H 4.61, N 8.22, F 13.81, Gd 13.45, S 1.37;

Found: C 37.87, H 4.70, N 8.22, F 13.90, Gd 13.48, 1.36%

WITH.

Example 17

a) 5- (Ethoxycarbonyl) pentyl-2,3,4,6-tetra-O-acetyl-α-D-manopyranoside

In an analogous manner to that described in the literature for the synthesis of arylglycopyranosides (J. Conchie and GA Lavva in Methods in Carbohydrate Chemistry (RL Whistler, ML Wolfrom and JN BeMiller, Eds.), Academic Press, New York, Vol. II, 90, p. 345-347 (1963)), a reaction of 156.2 g (400 mmol) of D-mannose pentaacetate as a mixture of α, β-anomers (α: β = 4: 1) [synthesis 1,2,3,4, 6-penta-O-acetyl-.alpha.,. Beta.-D-manopyranoses, see: ML Wolr and A. Thompson in Methods in Carbohydrate Chemistry (RL Whistler, ML Wolfrom and JN BeMiller, Eds.), Academic Press, New York, Vol. II, 53, p. 211-215 (1963)] with 67 ml (400 mmol) of ethyl 6-hydroxyhexanoate and 60.8 ml (520 mmol) of tin tetrachloride in a total of 600 ml of 1,2-dichloroethane. Purification by column chromatography on hexane-ethyl acetate (2: 1) gave the title compound as a colorless viscous oil (100.05 g, 51%). By ^X-H NMR spectrum, the product is solely the pure a-anomer.

analysis:

Calculated: C 52.94, H 6.77; Found:% C, 52.80;% H, 6.78.

b) 5- (Carboxy) pentyl-2,3,4,6-tetra-O-benzyl-α-D-manopyranoside

To a suspension of 141.0 g (289 mmol) of the title compound of Example 17a in 200 mL dioxane at room temperature and vigorous stirring was added portionwise 238.5 g (4.26 mmol) of finely powdered potassium hydroxide. An additional 200 mL of dioxane was added for better stirring and the resulting suspension was heated to boiling. At this temperature, a total of 372 ml (3.128 mol) of benzyl bromide was added dropwise over 2 hours. After 4 hours at 110 ° C and 12 hours at room temperature, the reaction mixture is slowly poured into 2.5 liters of ice water and the aqueous phase is extracted thoroughly with diethyl ether. The ether phase is washed, dried over sodium sulphate, filtered and the diethyl ether is evaporated in vacuo. Excess benzylbromic is quantitatively distilled off from the reaction mixture in an oil pump vacuum at an oil bath temperature of 180 ° C. The oily-oily residue obtained is purified by chromatography on silica gel (ethyl acetate / hexane 1:10). Yield: 172.2 g (91.0%) of the title compound as a colorless, highly viscous oil.

analysis:

Calculated: C 75.68, H 7.16;

Found:% C, 75.79;% H, 7.04.

c) 5- (Carboxy) pentyl-2,3,4,6-tetra-O-benzyl-α-D-mannopyranoside mmol) N-hydroxysuccinimide N-hydroxysuccinimide N-hydroxysuccinimide ester was added and total N-hydroxysuccinimide added. The mixture was cooled to 0 ° C (91.5 mmol) dicyclohexylcarbodiimide. The mixture is stirred at 0 ° C for 4 hours at room temperature under vacuum and 100 ml is added to the residue.

60.0 g (91.5) are dissolved in 750 ml of 10.4 g (91.5 mmol) and 18.9 g are added, stirred for 1 hour at room temperature. The solvent is ethyl acetate, after cooling to 0 ° C, the precipitated urea is filtered off. The filtrate was evaporated to dryness in vacuo, and the resulting oily-oily residue was purified by chromatography on silica gel with ethyl acetate-hexane (1:20) to give 61.23 g (89.0%) of the title compound as a colorless viscous oil.

analysis:

H, 6.57; N, 1.86. Found:% C, 70.39;% H, 5.64;% N, 1.91.

d) 2,6-Bis {6- _ε- 2- _alpha - [10aD-6-carbonylpentyl- (2,3,4,6-tetra-O-benzyl) mannopyranose] -L-lysine} methyl ester

To a solution of 4.26 g (18.30 mmol; 0.5 mol equivalents, based on carboxylic acid) of L-lysine methyl ester dihydrochloride (commercially available from Bachem) and 4.05 g (40.26 mmol) of triethylamine in 100 mL of dimethylformamide A solution of 27.51 g (36.6 mmol) of the title compound of Example 17c in 150 mL of DMF was added dropwise at 0 ° C. After the addition was complete, the mixture was stirred for an additional hour at 0 ° C and then overnight at room temperature. The solvent was evaporated in vacuo and the residue was taken up in 300 ml of ethyl acetate. The precipitated urea is filtered off and the filtrate is washed twice with 100 ml of 5% aqueous soda solution. The organic phase is dried over magnesium sulphate and evaporated to dryness in vacuo. Purification by chromatography on silica gel (n-hexane-isopropanol 25: 1) afforded the title compound (39.56 g, 75.4%) as a colorless oil.

analysis:

H, 7.31; N, 1.95.

Found:% C, 72.90;% H, 7.29;% N, 2.02.

e) 2,6-Bis ([6- _ε- 2-Ν _α - [10aD-6-carbonylpentyl- (2,3,4,6-tetra-O-benzyl) mannopyranose]] - L-lysine

30.0 g (20.92 mmol) of the compound prepared in Example 17d are dissolved in 150 ml of ethanol. To this solution was added a solution of 4 g (100.0 mmol) of sodium hydroxide in 10 ml of distilled water and stirred at 50 ° C for 3 hours. According to thin-layer chromatography, the saponification is quantitative. After evaporation in vacuo to dryness, the residue is taken up in 300 ml of ethyl acetate and the organic phase is extracted with dilute aqueous citric acid solution (2 x 100 ml). After drying over sodium sulfate, filtration and evaporation to dryness in vacuo, the residue is chromatographed on silica gel with n-hexane-isopropanol (13: 1). Yield 25.56 g (88.5%) of the title compound as a colorless oil.

analysis:

H, 7.31; N, 1.95.

Found:% C, 72.78;% H, 7.33;% N, 1.96.

f) 2,6-3is [6- _ε- 2-Ν _α - [1-O-α-D-6-carbonylpentyl- (2,3,4,6-tetra-O-benzyl) -manopyranose] -L- lysine] -N-hydroxysuccinimide ester

14.0 g (9.15 mmol) of the title compound of Example 17e are dissolved in 100 ml of dimethylformamide and a total of 1.04 g (9.15 mmol ·) of N-hydroxysuccinimide is added. The mixture was cooled to 0 ° C and dicyclohexylcarbodiimide (1.89 g, 9.15 mmol) was added. The mixture was stirred at 0 ° C for 1 hour and then at room temperature for 4 hours. The solvent was removed in vacuo and ethyl acetate (100 ml) was added. After cooling to 0 ° C, the precipitated urea is filtered off and the filtrate is evaporated to dryness in vacuo. The resulting oily-oily residue was purified by chromatography on silica gel with ethyl acetate-n-hexane (1:20). Thus, 12.94 g (92.4%) of the title compound are obtained as a colorless viscous oil.

analysis:

H, 7.05; N, 2.74.

Found:% C, 71.39;% H, 7.14;% N, 2.81.

g) 2,6-N, N'-Bis [1-O-α- (6-carbonyl) pentyl-2,3,4,6-tetra-O-benzyl-mannopyranose] -L-lysine] -1 , 7- (1,4,7-triazaheptane) diamide

To a solution of 0.47 g (4.57 mmol) of diethylenetriamine in 25 mL of dimethylformamide at 0 ° C was slowly added dropwise a solution of 14.0 g (9.15 mmol; 2 mol equivalents based on the amine used) of the title compound of Example 17f at 100. ml of dimethylformamide. After the addition, the mixture was stirred for an additional hour at 0 ° C and then overnight at room temperature. After evaporation in vacuo to dryness, the residue is taken up in 200 ml of ethyl acetate, the precipitated urea is filtered off and the filtrate is washed twice with 50 ml of 5% aqueous soda solution. The organic phase is dried over magnesium sulphate and evaporated to dryness in vacuo. The residue was chromatographed on silica gel with n-hexane-isopropanol (25: 1) to give 9.53 g (71.4%) of the title compound as a colorless oil.

analysis:

H, 7.42; N, 3.36.

Found:% C, 72.90;% H, 7.39;% N, 3.32.

h) 2-N- [2- (N-Ethyl-N-perfluorooctylsulfonyl) amino] acetyl-6-N- (benzyl oxycarbonyl) -L-lysine methyl ester

20.8 g (35.6 mmol) of 2- (N-ethyl-N-perfluorooctylsulfonyl) aminoacetic acid and 3.60 g (35.6 mmol) of triethylamine are dissolved in 200 ml of dimethylformamide and 4.09 g (35 mmol) are added. , 6 mmol) of N-hydroxysuccinimide. The mixture was cooled to 0 ° C and dicyclohexylcarbodiimide (7.34 g, 35.6 mmol) was added. The mixture was stirred at 0 ° C for 1 hour and then at room temperature for 4 hours. Cool to 0 ° C and add dropwise a solution of 11.77 g (35.6 mmol) of β-Ν-benzyloxycarbonyl-L-lysine-methyl ester hydrochloride and 4.0 g (40.0 mmol) of triethylamine in 100 mL over 10 min. dimethylformamide. The mixture was stirred at 0 ° C for 1 hour and then at room temperature overnight. The solvent was removed in vacuo and ethyl acetate (100 ml) was added. The precipitated urea is filtered off and the filtrate is washed twice with 100 ml of 5% aqueous soda solution. The organic phase is dried over magnesium sulphate and evaporated to dryness in vacuo. Zvy78

the shock clears chromatography on the silica gel in the system n-hexane-ethyl acetate (20: 1). It will be obtained so 27.43 g (88.0%) viscous oil. analysis: Calculated: 38.41% C, 3.45% H, 4.80% M, 36 .89%, E, 3.66% S; Found: 38.45% C, 3.38% H, N, 4.88 37.02 % F, 3.71% S.

i) _2-Ν α - {[2- (N-Ety1-N-perfluorooctylsulfonyl) -amino] -acetyl} _-6-Ν ε - (benzyloxycarbonyl) -L-lysine

25.0 g (28.55 mmol) of the compound prepared in Example 17h were dissolved in 150 ml of ethanol. To this solution was added a solution of 4 g (100.0 mmol) of sodium hydroxide in 10 ml of distilled water and stirred at 50 ° C for 3 hours. According to thin-layer chromatography, the saponification is quantitative. After evaporation in vacuo to dryness, the residue is taken up in 300 ml of ethyl acetate and the organic phase is extracted with dilute aqueous citric acid solution (2 x 100 ml). After drying over sodium sulfate, filtration and evaporation to dryness in vacuo, the residue is chromatographed on silica gel with n-hexane-isopropanol (10: 1). Yield 22.73 g (92.4%) of the title compound as a colorless oil.

analysis:

Calculated: C 37.64, H 3.28, N 4.88, E 37.49, S 3.72; Found: C 37.65, H 3.38, N 4.88, F 37.52, S 3.73.

j) 1,4,7-Triazaheptane-4- {2-N- [2- (N-ethyl-N-perfluorooctylsulfonyl) amino] acetyl-6-N-benzyloxycarbonyl} -L-lysine-amide-1,7- bis {2,6-N, N'-bis [1-O-cis-D- (5-carbonyl) pentyl-2,3,4,6-tetra-O-benzylmanopyranose] -L-lysine-diamide }

15.33 g (17.8 mmol) of the title compound of Example 17i and 1.80 g (17.8 mmol) of triethylamine are dissolved in 250 ml of dry tetrahydrofuran. After cooling the reaction solution to -15 ° C to -20 ° C, a solution of 4.92 g (35.6 mmol) of isobutyl chloroformate in 50 ml of dry tetrahydrofuran is slowly added dropwise with stirring at this temperature, so that the internal temperature does not exceed -10. After 15 minutes at -15 ° C, a solution of 52.0 g (17.8 mmol) of the title compound of Example 17g and 1.80 g (17.8 mmol) of triethylamine in 300 mL of dry tetrahydrofuran is slowly added dropwise at -20 ° C. After 1 hour at -15 ° C and 2 hours at room temperature, the reaction solution is evaporated to dryness in vacuo. The residue is taken up in 500 ml of ethyl acetate and extracted twice with 200 ml of saturated sodium bicarbonate solution and once with 200 ml of water. The organic phase is dried over sodium sulphate, filtered and evaporated in vacuo. The oily residue was purified on silica gel with ethyl acetate-n-hexane (1:20) to give 54.6 g (81.6%) of the title compound as a colorless, very viscous oil.

analysis:

Calculated: C 65.09, H 6.45, N 3.72, F 8.58, S 0.85;

Found:% C, 65.13;% H, 4.41;% N, 3.69.% F, 0.92%.

k) 1,4,7-Triazaheptane-4- (2-N- [2- (N-ethyl-N-perfluorooctylsulfonyl) amino] acetyl} -L-lysine-amide-1,7-bis {2,6- N, N'-bis [1-OaD- (5-carbonyl) -pentyl-mannopyranose] -L-lysine-diamide}

To a solution of 50.0 g (13.28 mmol) of the compound prepared in Example 17j in 500 mL of ethanol was added 4.0 g of Pearlman catalyst (20% Pd / C) and hydrogenated at room temperature and 1 atm pressure as long as no more hydrogen is consumed. The catalyst is filtered off with suction, washed thoroughly with ethanol (about 400 ml) and the filtrate is concentrated to dryness in vacuo. Yield 26.85 g (93.0%) of the title compound as a very viscous colorless oil.

analysis:

Calculated: C 45.85, H 6.35, N 6.44, F 14.86, S 1.47; Found: C 45.76, H 6.35, N 6.41, F 14.92, S 1.39.

1) 1,4,7-Triazaheptane-4- {2-N- [2- (N-ethyl-N-perfluorooctylsulfonyl) amino] acetyl} -6-N- [1,4,7-tris (carboxylatomethyl) - 10- (3-aza-4-oxo-5-methyl-5-yl-pentanoyl) -1,4,7,10-tetraazacyclododecane] -L-lysine-amide-1,7-bis {2,6-N , N'-bis [1-O-α- (5-carbonyl) pentyl-manopyranose] -L-lysine diamide}, gadolinium complex

5.54 g (8.8 mmol; 2.2 mol equivalents, based on the amine component of Example 17k) gadolinium complex of 10- (4-carboxy-1-methyl-2-oxo-3-azabutyl) -1,4 , 7,10-tetraazacyclododecane-1,4, 7-triacetic acid (described in patent application

DE 197 28 954 Cl in Example 31h) and 0.37 g (8.8 mmol) of anhydrous lithium chloride are dissolved in 60 ml of absolute dimethylsulfoxide with stirring at 40 ° C and a total of 1.01 g (8, 8 mmol) of N-hydroxysuccinimide and a solution of 1.84 g (4.0 mmol) of the title compound of Example 17k in 40 ml of absolute dimethylsulfoxide. After cooling to room temperature, 1.82 g (8.8 mmol) of N, N'-dicyclohexylcarbodiimide was added to the reaction solution, and the mixture was stirred at room temperature for 12 hours. Sufficient acetone is then added to the resulting slurry to completely precipitate the title compound, the precipitate is filtered off with suction, dried, mixed with a leader, filtered from insoluble dicyclohexylurea and the filtrate is desalted and freed from the low molecular weight components using an Amicon® YM-3 ultrafiltration membrane. cut off 3000 Da). The retentate was lyophilized to give 8.77 g (78.7%) of a colorless lyophilisate containing 4.43% water (Karl-Fischer).

Analysis (calculated on the anhydrous substance):

Calculated: C 43.98, H 5.97, N 7.54, F 11.59, Gd 5.64, S 1.15;

Found: C 43.97, H 6.02, N 7.62, F 11.61, Gd 10.18, 1.15%

WITH.

Example 18

a) 2-Na-6-Ns-Bis [1-O-α-D-carbonylmethyl-2,3,4,6-tetra-O-benzyl-manopyranose] -L-lysine-methyl ester

10.95 g (18.30 mmol) of 1-carboxymethyloxy-2,3,4,6-tetra-O-benzyl-α-D-manopyranoside (prepared as described in DE 197 28 954 C1) are dissolved in 150 ml Dimethylformamide and a total of 2.09 g (18.3 mmol) of N-hydroxysuccinimide was added to this solution. The mixture was cooled to 0 ° C and 3.78 g (18.3 mmol) of dicyclohexylcarbodiimide was added. The mixture was stirred at 0 ° C for 1 hour and then at room temperature for 4 hours. It is then cooled to 0 [deg.] C. and a solution of 2.13 g (9.15 mmol; 0.5 mol equivalents, based on the carboxylic acid used) of L-lysine methyl ester dihydrochloride, commercially available from Bachem), is added dropwise over 1 hour. 02 g (20.13 mmol) of triethylamine in 70 ml of dimethylformamide. The mixture was stirred for 1 hour at 0 ° C and then overnight at room temperature. After evaporation in vacuo to dryness, the residue is taken up in 300 ml of ethyl acetate, filtered from the precipitated urea and the filtrate is washed twice with 100 ml of 5% aqueous soda solution. The organic phase is dried over magnesium sulphate and evaporated to dryness in vacuo. The residue was chromatographed on silica gel using n-hexane-isopropanol (25: 1). Yield: 10.05 g (82.3%) of the title compound as a colorless oil.

analysis:

H, 6.79; N, 2.10.

Found:% C, 71.90;% H, 6.79;% N, 2.09.

b) 2-N _and -6-N _e -Bis [10α-D-carbonylmethyl-2,3,4,6-tetra-O-benzyl-manopyranose] -L-lysine g (11.23 mmol) of the methyl ester of Example 18a was saponification analogously to Example 17e. Yield: 13.89 g (93.6% of theory) of the title compound as a colorless viscous oil.

analysis:

H, 6.71; N, 2.12. Found:% C, 71.84;% H, 6.69;% N, 2.15.

c) 2-N _and N _£ 6-bis [LOAD-carbonylmethyl-2,3,4,6-tetra-0-benzyl-mannopyranose] -L-lysine-N-hydroxysuccinimide ester

12.09 o (9.15 mmol) of the title compound of Example 18b are dissolved in 100 ml of dimethylformamide and a total of 1.04 g (9.15 mmol) of N-hydroxysuccinimide is added. The mixture was cooled to 0 ° C and dicyclohexylcarbodiimide (1.89 g, 9.15 mmol) was added. The mixture was stirred at 0 ° C for 1 hour and then at room temperature for 4 hours. The solvent was evaporated in vacuo and 100 mL of ethyl acetate was added to the residue. It is then cooled to 0 ° C, the precipitated urea is filtered off and the filtrate is evaporated to dryness in vacuo. The resulting oily-oily residue was purified by chromatography on silica gel with ethyl acetate-n-hexane (1:25). Yield: 12.24 g (94.4% of theory) of the title compound as a colorless viscous oil.

analysis:

H, 6.47; N, 2.96.

Found:% C, 70.31;% H, 6.44;% N, 3.01.

d) 6-N-Benzyloxycarbonyl-2-N - {[2,6-N, N'-bis (10α-D-carbonylmethyl-2,3,4,6-tetra-O-benzyl-manopyranose)] - L-lysyl 1-L-lysine-1- (4-perfluorooctylsulfonyl) piperazine] amide

To a solution of 19.0 g (13.4 mmol) of the carboxylic acid N-hydroxysuccinimide ester prepared in Example 18c in 75 ml of DMF at 0 ° C was added dropwise a solution of 11.13 g (13.4 mmcl ·) of the title compound of Example 1c. in 50.0 ml of dimethylformamide, precooled to 0 ° C. The resulting reaction solution was stirred at 0 ° C for 2 hours and at room temperature for 12 hours. The precipitated cicyclohexylurea is filtered off and the solvent is evaporated to dryness in vacuo. The residue was chromatographed on silica gel with dichloromethane-ethanol (28: 1). Chromatography is performed using a solvent gradient, continuously increasing the proportion of polar solvent (ethanol) in the elution mixture. Yield: 25.28 g (88.4% of theory) of the title compound as a colorless, very viscous oil.

analysis:

Calculated: C 59.10, H 5.34, N 3.96, F 15.13, S 1.50; Found: C 59.18, H 5.35, N 4.02, F 15.15, S 1.56.

e) 2-N - {[2,6-N, N'-Bis (1-O-α-D-carbonylmethyl-manopyranose)] -L-lysyl-L-lysine- [1- (4-perfluorooctylsulfonyl) piperazine] amide

To a solution of 20.0 g (9.37 mmol) of the compound prepared in Example 18d in 200 mL of ethanol was added 1.5 g of Pearlman's catalyst (20% Pd / C) and hydrogenated at room temperature and 1 atm for so long. until hydrogen consumption stops. The catalyst was filtered off with suction, washed well with ethanol (2 x 100 ml) and the filtrate was evaporated to dryness in vacuo. Yield 11.62 g (97.0%) of the title compound as a very viscous colorless oil.

analysis:

H, 4.65; N, 6.57; F, 25.25; S, 2.51; Found: C, 38.46; H, 4.65; N, 6.51; F, 25.23; S, 2.52.

f) 6-N- [1,4,7-Tris (carboxylatomethyl) -10- (3-aza-4-oxo-5-methyl-5-ylpentanoyl) -1,4,7,10-tetraazacyclododecane] -2-N - {[2,6-N, N'-bis (1-O-α-D-carbonylmethyl-manopyranose)] - L-lysyl) -L-lysine- [1- (4-perfluorooctylsulfonyl) piperazine] amide , gadolinium complex

9.98 g (15.84 mmol; 2.2 mol equivalents based on the amine component of Example 18e) gadolinium complex 10- (4-carboxy-1-methyl-2-oxo-3-azabutyl) -1,4, 7,10-tetraazacyclododecane-1,4,7-triacetic acid (described in patent application

DE 197 28 954 Cl in Example 31h) and 0.67 g (15.84 mmol) of anhydrous lithium chloride are dissolved with stirring at 40 ° C in 100 ml of absolute dimethylsulfoxide and at this temperature a total of

1.82 g (15.84 mmol) of N-hydroxysuccinimide and 9.19 g (7.19 mmol) of the title compound of Example 18e in 50 ml of absolute dimethylsulfoxide. After cooling to room temperature, N, N'-dicyclohexylcarbodiimide (3.27 g, 15.84 mmol) was added to the solution and stirred at room temperature for 12 hours. Sufficient acetone is added to the resulting suspension to completely precipitate the title compound, the precipitate is filtered off with suction, dried, mixed with water, the insoluble dicyclchexylurea is filtered off with suction and the filtrate is filtered through an Amicon® YM-3 ultrafiltration membrane (cut off: 3000 Da). ) to desalinate and remove any low molecular weight components that may be present. The retentate is then lyophilized. 11.85 g (87.2%) of product are obtained in the form of a colorless lyophilisate which contains 5.54% of water (Karl-Fischer).

Analysis (calculated on the anhydrous substance):

Calculated: C 38.12, H 4.64, N 8.15, F 20.38, S 1.70, Gd 8.32;

Found: C, 38.16; H, 4.59; N, 8.18; F, 20.37; S, 1.68; Gd, 8.28.

Example 19

a) 1,7-Bis (benzyloxycarbonyl) -4- (3-oxa-2H, 2H, 4H, 4H, 5H, 5H-O-fluorotridecanoyl) -1,4,7,10-tetraazacyclododecane

To a solution of 10.75 g (24.4 mmol) of 1,7-bis (benzyloxycarbonyl) -1,4,7,10-tetraazacyclododecane in 150 ml of tetrahydrofuran and 15 ml of chloroform is added a solution of 12 at 0 DEG C. and under a nitrogen atmosphere. 74 g (24.4 mmol) of the title compound of Example 19g in 150 mL of tetrahydrofuran. A total of 18.0 g (36.6 mmol) of EEDQ (2-ethoxy-1-ethoxycarbonyl-1,2-dihydroquinoline) was then added portionwise at 0 ° C and the mixture was stirred overnight at room temperature. After evaporation in vacuo, the residual oil is chromatographed on silica gel with n-hexane-isopropanol (16: 1). 15.89 g (69.0% based on the charged secondary amine) of the monoamide and 3.8 g (8.8%) of the diamide are obtained as a by-product. The resulting compound is isolated as a colorless oil.

analysis:

H, 3.95; F, 34.19; N, 5.93.

Found: C 45.72, H 4.01, F 34.22, N 5.88%.

b) 1,7-Bis (benzyloxycarbonyl) -4- (3-oxa-2H, 2H, 4H, 4H, 5H, 5'-perfluorotecanoyl) -10- [1-SaD- (2-carbonyl)] ethyl 2,3,4,6-tetra-0-acetyl-mannopyranose] -1,4,7,10-tetraazacyclododecane

7.09 g (13.4 mmol) of 3- (2,3,4,6-tetra-O-acetyl-manopyranosyl) propionic acid N-hydroxysuccinimide ester (prepared according to: J. Haensler et al., Bioconjugate Chem. 4, 85 (1993);

Chipowski, S. and Lee, Y.C., (1973), Synthesis of 1-thioaldosides;

Carbohydrate Research 31, 339-346) was dissolved in 100 ml of dimethylformamide and a pre-cooled solution of 12.65 g (13.4 mmol) of the title compound of Example 19a in 100 ml of dimethylformamide was added dropwise at 0 ° C. The mixture was stirred at 0 ° C for 2 hours and then at room temperature for 12 hours. The solvent was evaporated to dryness in vacuo and the residue was chromatographed on silica gel with dichloromethane-ethyl acetate (20: 1) using a solvent gradient with a continuous increase in the proportion of ethyl acetate in the mixture. Yield: 16.23 g (88.9% of theory) of the title compound as a colorless, very viscous oil.

analysis:

Calculated: C 46.70, H 4.36, N 4.11, F 23.69, S 2.35; Found:% C, 46.66;% H, 4.35;% N, 4.12;% F, 23.65.

c) 1- ^(3-Oxa-R 2B, 2H, 4H, 4H, 5H, 5H-perfluorotridecanoyl) -7- [1-SaD- (2-carbonyl) -ethyl-2,3,4,6-tetra-O -acetylmanopyranóza] -1,4,7,10tetraazacyklododekán

To a solution of 15.0 g (11.0 mmol) of the compound prepared in Example 19b in 150 mL of ethanol was added 1.0 g of Pearlman catalyst 86 (20% Pd / C) and hydrogenated at room temperature and 1 atm pressure until hydrogen consumption stops. The catalyst was filtered off with suction, washed thoroughly with ethanol (2 x 75 ml) and the filtrate was evaporated to dryness in vacuo. Yield 11.56 g (96.0%) of the title compound as a very viscous colorless oil.

analysis:

Calculated: C 40.59, H 4.33, N 5.12, F 29.50, S 2.93; Found:% C, 40.63;% H, 4.35;% N, 5.11;% S, 2.52%.

d) 1- (3-Oxa-2H, 2H, 4H, 4H, 5H, 5H-perfluoridecanoyl) -7- [1-SaD- (2-carbonyl) ethyl-mannopyranose] -1,4,7, 10-tetraazacyclododecane

The resulting compound of Example 19c (10.0 g, 9.13 mmol) was suspended in 100 mL of absolute methanol and a catalytic amount of sodium methoxide was added at 5 ° C. After 3 hours at room temperature, TLC analysis (eluent: chloroform-methanol 4: 1) showed a quantitative conversion. The resulting clear reaction solution was neutralized by addition of Amberlite IR 120 (H ⁺ form), the ion exchanger was filtered off with suction, washed with methanol and the methanolic filtrate was evaporated to dryness in vacuo. The residue was subjected to silica gel column chromatography using dichloromethane-n-hexane-ethyl acetate (15: 20: 1) as the solvent gradient using a continuous increase in ethyl acetate in the elution mixture. According to the size of interaction constants (Ji, ₂ =

0.9 Hz) in the &lt; ³ &gt; H-NMR spectrum can be clearly inferred from the [alpha] -configuration at the anomeric center of D-manopyranose. This .alpha.-configuration is present exclusively, i.e. the amount of any formed anomer with the .beta.-configuration lies below the detection threshold of ¹ H-NMR spectroscopy. Yield: 8.28 g (98.0%) of the title compound as a colorless, very viscous oil.

analysis:

Calculated: C 37.59, H 4.24, N 6.05, F 34.85, S 3.46;

Found: C 37.57, H 4.28, N 6.02, F 34.85, S 3.44.

e) 1- (3-Oxa-2H, 2H, 4H, 4H, 5H, 5H-perfluorotridecanoyl) -7- [1-SaD- (2-carbonyl) ethyl-mannopyranose] - 4,10 bis- [1,4] 7-tris (methyl carboxylate) -10- (3-aza-4-oxo-5-methyl-5-ylpentanoyl)] - 1,4,7,10-tetraazacyclododecane, digadolinium complex

2.48 g (3.94 mmol; 4.4 mol equivalents based on the charged diamine component of Example 19d) of the gadolinium complex of 10- (4-carboxy-1-methyl-2-oxo-3-azabutyl) -1,4, 7, 10-tetraazacyclododecane-1,4,7-triacetic acid (described in patent application

DE 197 28 954 Cl in Example 31h) and 167 mg (3.94 mmol) of anhydrous lithium chloride are dissolved with stirring at 40 ° C in 40 ml of absolute dimethylsulphoxide and at this temperature 453 mg (3.94 mmol) of N- are added. n-hydroxysuccinimide and 890 mg (0.895 mmol) of the title compound of Example 19d in 10 mL of absolute dimethylsulfoxide. After cooling to room temperature, 814 mg (3.946 mmol) of N, N'-dicyclohexylcarbodiimide was added to the solution and stirred for 12 hours at room temperature. Sufficient acetone is added to the resulting suspension to completely precipitate the resulting compound, the precipitate is suctioned off, dried, mixed with water, the insoluble dicyclohexylurea is suctioned off and the filtrate is filtered through an Amicon® YM-3 ultrafiltration membrane (cut off: 3000 Da). to desalinate and remove the low molecular weight components. The retentate is then lyophilized. 1.32 g (69.1%) of product are obtained as a colorless lyophilisate containing 7.65% water (Karl-Fischer).

Analysis (calculated on the anhydrous substance):

Calculated: C 37.43, H 4.45, N 9.12, F 15.02, S 1.49, Gd 14.63;

Found: C, 37.42; H, 4.50; N, 9.18; F, 15.07; S, 1.51; Gd, 14.58.

f) 3-oxa-2H, 2H, 4H, 4H, 5H, 5H-perfluorotridecanoic acid tert-butyl ester

25.0 g (53.8 mmol) of 1H, 1H, 2H, 2H-perfluoro-1-decanol (commercially available from Lancaster) are dissolved in 250 ml of absolute toluene and a catalytic amount (about 0.75) is added at room temperature. g) tetra-n-butylammonium hydrogen sulfate. A total of 7.55 g (134.6 mmol; 2.5 equivalents, based on the alcohol component used) of finely powdered potassium hydroxide is then added at 0 ° C, followed by 15.73 g (80.7 mmol; 1.5 equivalents). based on the alcohol component used) of bromoacetic acid tert-butyl ester. The mixture was stirred at 0 ° C for 2 hours and then at room temperature for 12 hours. The resulting reaction solution is quenched with a total of 500 ml of ethyl acetate and 250 ml of water, the organic phase is separated and washed twice with water, dried over sodium sulphate, filtered and the solvent is evaporated off under vacuum. The oily residue was then purified on silica gel with ethyl acetate-hexane (1:10). Yield: 26.3 g (84.6%) of the title compound as a colorless, very viscous oil.

analysis:

% H, 2.61;% F, 55.85;

Found:% C, 33.29;% H, 2.61;% F, 55.90.

(g) 3-Oxa-2H-2H, AH, 4H, 5H, 5H-perfluorotridecanoic acid

20.0 g (34.58 mmol) of the title compound of Example 19f are suspended in 200 ml of a 2: 1 mixture of methanol and 0.5 M sodium hydroxide at room temperature and the suspension is heated to 60 ° C. After 12 hours at 60 ° C, the clear reaction solution is neutralized with cation exchange resin Amberlite IR 120 (H ⁺ -form), the ion exchanger is filtered off with suction and the resulting aqueous methanolic filtrate is evaporated to dryness in vacuo. The amorphous-oily residue was then purified by chromatography on silica gel with ethyl acetate-n-hexane (1: 3). Yield 16.0 g (88.6%) of a colorless, very viscous oil.

analysis:

H, 1.35; F, 61.85.

Found: C, 27.58; H, 1.36; F, 61.90.

Example 20

a) 6-Benzyloxycarbonyl-2- [2- (N-ethyl-N-perfluorooctylsulfonyl) amino] acetyl-L-lysine methyl ester

To a solution of 8.0 g (24.4 mmol) of ε-carbonyloxybenzyl-L-lysine methyl ester hydrochloride (commercially available from Bachem) in a mixture of 150 mL of tetrahydrofuran, 15 mL of chloroform and 2.62 g (26.0 mmol) of triethylamine A solution of 16.18 g (27.0 mmol) of 2- (N-ethyl-N-perfluorooctylsulfonyl) aminoacetic acid (prepared according to DE 196 03 033) in 50 ml of tetrahydrofuran is added dropwise at 0 ° C under nitrogen. A total of 18.0 g (36.6 mmol) of EEDQ (2-ethoxy-1-ethoxycarbonyl-1,2-dihydroquinoline) was then added portionwise at 0 ° C and the mixture was stirred overnight at room temperature. After evaporation in vacuo, the oily residue is chromatographed on silica gel with n-hexane-isopropanol (15: 1). Yield: 17.0 g (79.6% based on the primary amine used) of the title compound as a colorless oil.

analysis:

Calculated: C 38.41, H 3.45, E 36.89, N 4.80, S 3.66; Found: C 38.42, H 3.47, F 36.92, N 4.87, S 3.64.

b) 2- [2- (N-Ethyl-N-perfluorooctylsulfonyl) amino] acetyl-L-lysine methyl ester

To a solution of 15.0 g (20.23 mmol) of the compound prepared in Example 20a in 200 mL of ethanol was added 800 mg of Pearlman's catalyst (20% Pd / C) and hydrogenated to consume the calculated amount of hydrogen. The catalyst is filtered off with suction, washed thoroughly with ethanol and the filtrate is evaporated to dryness in vacuo. Yield 14.68 g (97.9%) of the product as a very viscous colorless oil.

analysis:

H, 3.26; F, 43.56; N, 5.67; S, 4.32;

Found: C 32.42, H 3.27, F 43.60, N 5.67, S 4.34.

c) β- (1-O-α-D-Carbonylmethyl-2,3,4, β-tetra-O-benzylmanopyranose) -2- [2- (N-ethyl-N-perfluorooctylsulfonyl) amino] acetyl-L-lysine-methyl ester

21.31 g (35.6 mmol) of 1-carboxymethyloxy-2,3,4,6-tetra-O-benzyl-α-D-manopyranoside (prepared according to DE 197 28 954 C1) and 3.60 g (35, 6 mmol) of triethylamine is dissolved in 500 ml of dry tetrahydrofuran. The reaction solution was cooled to -15 to -20 ^C and stirred thereto at this temperature, slowly, a solution of 4.92 g (35.6 mmol) of isobutyl chloroformate in 75 mL of dry THF rate such that the internal temperature does not exceed -10 ° C. After the addition, the mixture was kept at -15 ° C for 15 minutes and then a solution of 26.39 g (35.6 mmol) of the title compound of Example 20b and 3.60 g (35.6 mmol) triethylamine was slowly added dropwise at -20 ° C. in 100 ml dry tetrahydrofuran. After 1 hour at -15 ° C and 2 hours at room temperature, the reaction solution is evaporated to dryness in vacuo. The residue is taken up in 250 ml of ethyl acetate and the solution is washed with 2 x 100 ml of saturated sodium bicarbonate solution and then once with 200 ml of water. After drying over sodium sulfate, the solution was filtered and the ethyl acetate was evaporated in vacuo. The oily residue was purified on silica gel with ethyl acetate-hexane (1:10). 38.12 g (81.0%) of the title compound are obtained as a colorless, very viscous oil.

analysis:

Calculated: C 49.92, H 3.92, N 2.53, F 29.19, S 2.90; Found: 49.99% C, 4.11% H, 2.69% N, 29.22% F, 3.01% ^S.!

d) 6- (1-O-α-D-Carbonylmethyl-2,3,4, β-tetra-O-benzylmanopyranose) -2- [2- (N-ethyl-N-perfluorooctylsulfonyl) amino] acetyl-L-lzine

27.65 g (20.92 mmol) of the compound prepared in Example 20c are dissolved in 250 ml of ethanol. To this solution was added a solution of 4.0 g (100.0 mmol) of sodium hydroxide in 10 ml of distilled water and stirred at 50 ° C for 3 hours. Thin layer chromatography shows that the saponification of the methyl ester is quantitative after this time.

After evaporation in vacuo to dryness, the residue is taken up in 300 ml of ethyl acetate and the organic phase is extracted with dilute aqueous citric acid solution (2 x 100 ml). After drying over sodium sulfate, filtration and evaporation to dryness in vacuo, the residue is chromatographed on silica gel with n-hexane-chloroform-isopropanol (15: 10: 1). Yield 24.31 g (88.9%) of the title compound as a colorless viscous oil.

analysis:

Calculated: C 51.46, H 4.70, M 3.21, F 24.71, S 2.45; Found: C 51.49, H 4.71, N 3.19, F 24.72, S 2.41.

e) 6- (1-O-α-D-Carbonyl-methyl-mannopyrano) -2- [2- (N-ethyl-N-perfluorooctylsulfonyl) amino] acetyl-L-lysine

20.0 g (15.30 mmol) of the title compound of Example 20d are dissolved in a mixture of 250 ml of 2-propanol and 25 ml of water and 1.0 g of palladium catalyst (10% Pd / activated carbon) is added. Hydrogenate for 12 hours at room temperature and 1 atm. The catalyst is then filtered off and the filtrate is evaporated to dryness in vacuo. The residue was dissolved in 200 ml of methanol and the reaction product was separated by the addition of 800 ml of diethyl ether. The product is filtered off with suction and dried under vacuum at 50 ° C. Yield 14.32 g (99.0%) of an amorphous solid.

analysis:

Calculated: C 35.56, H 3.84, N 4.44, S 3.39, F 34.15; Found: C, 35.58; H, 3.81; N, 4.45; S, 3.40; F, 34.17.

f) 6- (1-Oa-D-Carbonylmethyl-manopyranose) -2- [2- (N-ethyl-N-perfluorooctylsulfonyl) amino] acetyl-L-lysine-N- (2-hydroxyprop-3-yl- [1, 2-hydroxyprop-3-yl] - [1,3] 4,7-tris (carboxylatomethyl) -1,4,7,10-tetraazacyclododecan-10-yl]} amide, gadolinium complex

The resulting compound of Example 20e (7.48 g, 7.91 mmol) was dissolved in dimethyl sulfoxide (50 mL) at 40 ° C and N-hydroxysuccinimide (1.00 g, 8.70 mmol) was added. The mixture was cooled to 20 ° C, dicyclohexylcarbodiimide (1.795 g, 8.7 mmol) was added and stirred at 20 ° C for one hour and then at 40 ° C for 4 hours. A solution of 4.53 g (7.91 mmol) of the gadolinium complex of 10 - (2-hydroxy-3-aminopropyl) -4,7,10-tris (carboxymethyl) -1,4 is added dropwise at this temperature over 10 minutes. 7,10-tetraazacyclododecanine (for preparation see WO 97/02051) in 20 ml of dimethylsulfoxide. The mixture was stirred at 40 ° C for 1 hour and then at room temperature overnight. Sufficient acetone is added to the resulting suspension to completely precipitate the title compound, the precipitate is filtered off with suction, dried, stirred with water, the insoluble dicyclohexylurea is filtered off with suction and the filtrate is filtered through an Amicon® YM-3 ultrafiltration membrane (cut off: 3000 Da). to desalinate and remove the low molecular weight components. The retentate is then lyophilized. 9.71 g (81.7%) of product are obtained as a colorless lyophilisate containing 3.97% water (Kari-Fischer).

Analysis (calculated on the anhydrous substance):

Calculated: C 35.16, H 4.16, N 7.45, F 21.48, Gd 10.46, S 2.13;

Found: C 35.17, H 4.20, N 7.42, F 21.49, Gd 10.48, 2.09%

WITH.

Example 21

a) 6-N- [1-OaD- (Ξ-Carbonyl) pentyl-2,3,4,6-tetra-O-benzyl-manopyranose] -2-N- [2- (N-ethyl-N-perfluorooctylsulfenyl) ) amino] acetyl-L-lysine methyl ester

5.23 g (8.0 mmol) of 5- (carboxy) pentyl-2,3,4,6-tetra-O-benzyl-α-D-manopyranoside (described in Example 10c), 1.3 g (8, 0 mmol) of 1-hydroxybenzotriazole and 2.6 g (8.0 mmol) of 2- (1H-benzotriazol-1-yl) -1,1,3,3-tetramethyluronium tetrafluoroborate (TBTU; Peboc Limited, UK) were dissolved in 75 ml of dimethylformamide and stirred for 15 minutes. 5.16 ml (30 mmol) of N-ethyldiisopropylamine and 5.93 g (8.0 mmol) of the amine prepared in Example 20b were added and the mixture was stirred at room temperature for 1.5 days. The solvent was evaporated to dryness in vacuo and the residue was chromatographed on silica gel with dichloromethane-ethyl acetate (30: 1) using a solvent gradient with a continuous increase in the ethyl acetate portion of the eluent. This gave 9.70 g (88.0%) of the title compound as a colorless, very viscous oil.

analysis:

Calculated: C 52.29, H 4.97, N 3.05, F 23.43, S 2.33; Found: C 52.33, H 4.95, N 3.12, F 23.50, S 2.30.

b) 6-N- [1-O-α- (5-Carbonyl) pentyl-2,3,4,6-tetra-O-benzylmanopyranose] -2-N- [2- (N-ethyl-N-perfluorooctylsulfonyl) ) amino] acetyl-L-lysine

9.0 g (12.40 mmol) of the title compound of Example 21a are dissolved in 150 ml of ethanol. To this solution was added a solution of 2.48 g (62.0 mmol) of sodium hydroxide in 15 ml of distilled water and stirred at 50 ° C for 3 hours. Thin layer chromatography shows the saponification of the methyl ester quantitatively. After evaporation in vacuo to dryness, the residue is taken up in 300 ml of ethyl acetate and the organic phase is extracted with dilute aqueous citric acid solution (2 x 100 ml). After drying over sodium sulfate, filtration and evaporation to dryness in vacuo, the residue is chromatographed on silica gel with n-hexane-chloroform-isopropanol (25: 10: 1). Yield 15.88 g (93.9%) of the title compound as a colorless, very viscous oil.

analysis:

Calculated: C 51.95, H 4.88, N 3.08, F 23.67, S 2.35; Found:% C, 51.99;% H, 4.91;% N, 3.09;% F, 23.70.

c) 6-N- [1-O-α-D- (5-Carbonyl) pentyl-manopyranose] -2-N- [2- (N-ethyl-N-perfluorooctylsulfonyl) amino] acetyl-L-lysine

13.0 g (9.52 mmol) of the title compound of Example 21b are dissolved in a mixture of 150 ml of 2-propanol and 25 ml of water and 1.0 g of palladium catalyst (10% Pd / C) is added. The mixture was hydrogenated at 1 atm at room temperature for 12 hours. The catalyst is filtered off and the filtrate is evaporated to dryness in vacuo. The residue is chromatographed on silica gel with n-hexane-chloroform-isopropanol (15: 10: 1). Yield: 9.09 g (95.1%) of the title compound as a colorless, very viscous oil.

analysis:

Calculated: 37.10% Found: 37.09% C,

C, 4.22; H, 4.19; F, 32.18; S, 3.10; H, 4.21; N, 4.19; F, 32.20; S, 3.13%.

d) 6-N- [1-OaD- (5-Carbonyl) pentyl-manopyranose] -2-N- [2- (N-ethyl-N-perfluorooctylsulfonyl) amino] acetyl-L-lysine-N- (2- hydroxyprop-3-yl- [1,4,7-tris (carboxylatomethyl) -1,4,7,10-tetraazacyclododecan-10-yl]} amide, gadolinium complex

The resulting compound of Example 21c (7.93 g, 7.91 mmol) was dissolved in dimethylsulfoxide (75 mL) at 40 ° C and N-hydroxysuccinimide (1.00 g, 8.70 mmol) was added. The mixture was cooled to room temperature, dicyclohexylcarbodiimide (1.795 g, 8.7 mmol) was added and stirred at 20 ° C for one hour and then at 40 ° C for 4 hours. Then a solution of 4.53 g (7.91 mmol) of the gadolinium complex of 10- (2-hydroxy-3-aminopropyl) -4,7,10-tris (carboxymethyl) is added dropwise at 40 ° C over 10 minutes. -1,4,7,10-tetraazacyclododecanine (preparation cf. WO 97/02051) in 20 ml of dimethylsulfoxide. The mixture was stirred at 40 ° C for one hour and then at room temperature overnight. Sufficient acetone-2-propanol (2: 1) was added to the resulting slurry until the title compound completely precipitated. The precipitate is aspirated, dried, mixed with water, the insoluble dicyclohexylurea is aspirated, and the filtrate is filtered through an Amicon® YM-3 ultrafiltration membrane (cut off: 3000 Da) to desalinate and remove the low molecular weight components. The retentate is then lyophilized. 9.71 g (78.8%) of the product are obtained in the form of a colorless lyophilisate containing 6.65% of water (Karl-Fischer).

Analysis (calculated on the anhydrous substance):

Calculated: C 36.97, H 4.52, N 7.19, F 20.71, Gd 10.03, S 2.06;

Found: C 37.02, H 4.50, N 7.22, F 20.69, Gd 10.08, 2.09%

WITH.

Example 22

a) 6-N- (4- [2,3-Bis (N, N-bis (tert-butyloxycarbonylmethyl) amino) propyl) phenyl} -3-oxapropionyl-2-N- (Ι-α-D-carbonylmethyl- manopyranose) -L-lysine- [1- (4-perfluorooctylsulfonyl) piperazine] amide

5.25 g (7.72 mmol) of 1- (4-carboxymethoxybenzyl) -EDTA tetra-tert-butyl ester (see U.S. Pat. No. 4,622,220) and 781 mg (7.72 mmol) of triethylamine are dissolved in 50 ml of methylene chloride. To this solution at -15 ° C was added dropwise a solution of 1.16 g (8.5 mmol) of isobutyl chloroformate in 10 ml of methylene chloride over 5 minutes, and the mixture was stirred at -15 ° C for another 20 minutes. The solution was cooled to -25 ° C and a solution of 7.07 g (7.72 mmol) of the title compound of Example 10e and 2.12 g (21.0 mmol) of triethylamine in 70 mL of tetrahydrofuran was added dropwise over 30 minutes. . Stirring is continued for 30 minutes at -15 ° C and overnight at room temperature. The solvent was distilled off in vacuo and the oily residue was taken up in 250 ml of chloroform. The chloroform phase is extracted with 2 x 100 ml of 10% aqueous ammonium chloride solution, dried over magnesium sulphate and evaporated to dryness in vacuo. Chromatography on silica gel using methylene chloride-ethanol (20: 1) afforded 9.60 g (79.0%) of a colorless, very viscous oil.

A.na ski:

Calculated: C 46.39, H 5.55, N 5.32, F 20.45, S 2.03; Found: C, 46.42, H, 5.51, N, 5.29, F, 20.49, S, 2.09.

b) 6-N- (4- [2,3-Bis (N, N-bis (carboxymethyl) amino) propyl] phenyl} -3-oxapropionyl-2- [1- (1-α-carbonylmethyl-mannopyranose) -L) -lysine- [1- (4-perfluorooctylsulfonyl (piperazine) amide)

9.0 g (5.70 mmol) of the title compound of Example 22a were dissolved in 150 mL of methanol. To this solution was added a solution of 4.0 g (100.0 mmol) of sodium hydroxide in 25 mL of distilled water and stirred at 60 ° C for 6 hours. The point of thin-layer chromatography is the quantification of the saponification of the tetra-tert-butyl ester after this time. After evaporation in vacuo to dryness, the residue is taken up warmly in 50 ml of dimethylsulfoxide and so much acetone-ethyl acetate (1: 1) is added until the resulting compound precipitates completely. The precipitate formed is filtered off with suction, washed well with ethyl acetate, dried, stirred with water and the solution is adjusted to pH 3.5 by addition of 1 M hydrochloric acid. After filtering off any insoluble matter, the filtrate is desalted and freed from the low molecular weight components by filtration on an Amicon® YM-3 ultrafiltration membrane (cut off: 3000 Da). The retentate is then lyophilized. Thus 6.76 g (87.6%) of the product are obtained in the form of a colorless lyophilizate containing 3.30% water (Karl-Fischer).

Analysis (calculated on the anhydrous substance):

Calculated: C 39.89, H 4.09, N 6.20, F 23.84, S 2.37; Found: C 39.92, H 4.15, N 6.22, F 23.92, S 2.29.

c) 6-N- {4- [2,3-Bis (N, N-bis (carboxylatomethyl) amino) propyl] phenyl} -3-oxapropionyl-2-N- (1-α-carbonylmethyl-manopyranose) -L -lysine- [1- (4-perfluorooctylsulfonyl) piperazine] amide, η-complex, disodium salt

3.0 g (2.22 mmol) of the title compound of Example 22b was dissolved in boiling in 150 ml of water-ethanol (3: 1) and 0.25 g (2.22) was added portionwise at 80 ° C. mmol) of manganese carbonate and the reaction solution thus obtained is refluxed for 5 hours. After cooling to room temperature, the solvent is removed in vacuo and the residue is dissolved in 200 ml of distilled water and n-butanol (1: 1). It is then adjusted to pH 7.2 with vigorous stirring by the addition of 1 N sodium hydroxide. After the n-butanol was completely removed in vacuo, the remaining aqueous phase was desalted and freed from the low molecular weight fractions by filtration through an Amicon® YM-3 ultrafiltration membrane (cut off: 3000 Da) and the retentate was lyophilized. 3.19 g (99.0%) of product are obtained as a colorless lyophilisate containing 5.08% water (Karl-Fischer).

Analysis (calculated on the anhydrous substance):

Calculated: C 37.23, H 3.54, F 22.25, M 3.73, N 5.79, Na, 17.17, S, 2.21;

Found: C, 37.30; H, 3.49; F, 22.29; M, 3.81; N, 5.76; Na, 3.19;

Example 23

a) 3-N-Benzyloxycarbonylaminoglutaric acid [1- (4-perfluorooctylsulfonyl) piperazine] monoamide

To a stirred solution of 25.0 g (94.96 mmol) of 3- (N-benzyloxycarbonylamino) glutaric anhydride (for synthesis see Haranaka, Minoru; Yamamoto, Yu-ichi; Nitta, Hajime; Ishimaru, Toshiyasu; Teleay; Tetrahedron Lett., EN, 22; 39; 1981; 3833-3886) in 150 ml of absolute tetrahydrofuran is added dropwise a solution of 53.97 g (95.0 mmol) of 1-perfluorooctylsulfonylpiperazine in 150 ml of terrahydrofuran and the resulting reaction solution is refluxed for 12 hours. After cooling to room temperature, it is evaporated to dryness and the oily residue is purified on silica gel with dichloromethane-2-propanol (20: 1). Yield: 75.80 c (96.0%) of the title compound as a colorless viscous oil.

analysis:

Calculated: C 36.11, H 2.67, N 5.05, S 3.86, F 38.84; Found: C, 36.12; H, 2.61; N, 5.08; S, 3.88; F, 38.82.

b) [1- (4-perfluorooctylsulfonyl) piperazine] monoamide

3-aminoglutaric

31.50 g (37.88 mmol) of the title compound of Example 23b are dissolved in 300 ml of ethanol and 2.5 g of Pearlman catalyst (20% Pd / C) are added. The mixture was hydrogenated at 1 atm until quantitative hydrogen uptake. The catalyst was filtered off with suction, washed with ethanol and the filtrate was evaporated to dryness in vacuo. Yield: 25.22 g (95.5% of theory) of the title compound as a white yellow viscous oil.

analysis:

Calculated: C 29.28, H 2.31, N 6.03, S 4.06, F 46.31; Found: C 29.32, H 2.29, N 6.08, S 4.08, F 46.28.

c) 3-N- (1-α-D-carbonylmethyl-2,3,4,6-tetra-O-benzylmanopyranose) aminoglutaric acid [1- (4-trifluorooctylsulfonyl) piperazine] monoamide

21.52 g (18.96 mmol) of 1-carboxymethyloxy-2,3,4,6-tetra-O-benzyl-α-D-manopyranoside (for preparation see DE 197 28 954 C1) are dissolved in 100 ml at room temperature. of absolute dimethylformamide and to the solution were added 2.56 g (22.2 mmol) of N-hydroxysuccinimide at 0 ° C followed by 4.55 g (22.2 mmol) of dicyclohexylcarbodiimide. After 60 minutes at 0 ° C and 3 hours at 22 ° C, insoluble dicyclohexylurea is filtered off and the clear active ester solution thus obtained is slowly added dropwise at 0 ° C to a stirred solution of 13.22 g (18.96 mmol) of the compound of Example 23b in 100 ml of dimethylformamide. After 12 hours at room temperature, the solvent is removed in vacuo and the residue is stirred with 300 ml of ethyl acetate. The urea is filtered off and the organic filtrate is washed twice with 100 ml of saturated sodium bicarbonate solution, once with 100 ml of 10% aqueous citric acid solution and once with 200 ml of water. After drying the organic phase over sodium sulfate and filtration, ethyl acetate is evaporated in vacuo. The oily residue was purified by chromatography on silica gel with ethyl acetate / n-hexane (1:15). Yield: 21.39 g (88.3% of theory) of the title compound as a colorless, very viscous oil.

analysis:

Calculated: C 49.81, H 4.10, N 3.29, F 25.27, S 2.51; Found: C 49.89, H 4.11, N 3.32, F 25.22, S 2.51.

d) 3-N- (1-α-D-carbonylmethyl-manopyranose) glutaric acid [1- (4-perfluorooctylsulfonyl) piperazine] monoamide

19.55 g (15.30 mmol) of the title compound of Example 23c are dissolved in a mixture of 250 ml of 2-propanol and 25 ml of water and 1.5 g of palladium catalyst (10% Pd / activated carbon) are added. Hydrogenate for 12 hours at room temperature and pressure asm. The catalyst is then filtered off and the filtrate is evaporated to dryness in vacuo. The residue is dissolved in 200 ml of methanol and the reaction product is separated by the addition of a total of 800 ml of diethyl ether. The product is suctioned off and dried under vacuum at 40 ° C. Yield 17.49 g (97.5%) of an amorphous solid.

analysis:

Calculated: C 32.73, H 3.08, N 4.58, S 3.49, F 35.20; Found: C, 32.68; H, 3.15; N, 4.55; S, 3.50; F, 35.17.

e) 5-N- {2-hydroxyprop-3-yl- [1,4,7-] 3-N- (1-α-carbonylmethyl-manopyranose) giutar- [1- (4-perfluorooctylsulfonyl) piperazine] -amide tris (carboxylatomethyl) -1,4,7,10-tetraazacyclododecan-10-yl]} amide, gadolinium complex

14.43 g (15.84 mmol) of the title compound of Example 23d and 0.67 g (15.84 mmol) of anhydrous lithium chloride were dissolved in 100 ml of absolute dimethylsulfoxide at 40 ° C with stirring and a total of 1.82 g (15.84 mmol) of N-hydroxysuccinimide and a solution of 9.08 g (15.84 mmol) of gadolinium complex 10- (2-hydroxy-3-aminopropyl) -4,7,10-tris (carboxymethyl) - 1,4,7,10-tetraazacyclododecanine (preparation cf. WO 97/02051) in 50 ml dimethylsulfoxide. After cooling to room temperature, N, N'-dicyclohexylcarbodiimide (3.27 g, 15.84 mmol) was added to the reaction solution, and the mixture was stirred at room temperature for 12 hours. Sufficient acetone is added to the resulting suspension to completely precipitate the title compound, the precipitate is filtered off with suction, dried, stirred with water, the insoluble dicyclohexylurea is filtered off with suction and the filtrate is filtered through an Amicon® YM-3 ultrafiltration membrane (cut off: 3000 Da). to desalinate and remove any low molecular weight components that may be present. The retentate is then lyophilized. 18.71 g (80.2%) of product are obtained in the form of a colorless lyophilisate containing 4.87% of water (Karl-Fischer).

Analysis (calculated on the anhydrous substance):

Calculated: C 34.24, H 3.83, N 7.61, F 21.92, S 2.18, C-d 10.67;

Found: C 34.26, H 3.79, N 7.58, F 21.87, S 2.18, Gd 10.68.

Example 24

a) 1,7-Bis (benzyloxycarbonyl) -4- (3-oxapentane-1,5-dicarboxylic acid-1-oyl-5- (1- (4-perfluorooctylsulfonyl) piperazine) amide) -10- [2,6] -N, N'-bis (10aD-carbonylmethyl-2,3,4,6-tetra-O-benzyl-manopyranose)] - L-lysyl-1,4,7,10-tetraazacyclododecane

To a solution of the secondary amine prepared in Example 15a (27.0 g, 24.4 mmol) in tetrahydrofuran (150 mL) and chloroform (15 mL) at 0 ° C under nitrogen was added the title compound (33.04 g, 25.0 mmol). of Example 18c in 250 mL of tetrahydrofuran. A total of 18.0 g (36.6 mmol) of EEDQ (2-ethoxy-1-ethoxycarbonyl-1,2-dihydroquinoline) was then added portionwise at 0 ° C and the mixture was stirred overnight at room temperature. After evaporation in vacuo to dryness, the obtained residue is chromatographed on silica gel with n-hexane-isopropanol (25: 1). Yield: 45.87 g (78.0% based on charged secondary amine) of the title compound as a colorless oil.

101

Analysis:

Calculated: C 59.30, H 5.39, F 13.40, N 4.65, S 1.33. Found: C 59.32, H 5.37, F 13.37, N 4.70, S 1.34.

b) 1- {3-Oxapentane-1,5-dicarboxylic acid-1-oyl-5- [1- (4-perfluorooctylsulfonyl) piperazine] amide) -7- [2,6-N, N'-bis- ( 1-O-α-D-carbonylmethyl-manopyranose)] - L-lysyl-1,4,7,10-tetraazacyclododecane

The resulting compound of Example 24a (24.1 g, 10.0 mmol) was dissolved in ethanol (250 mL) and Pearlman's catalyst (20% Pd / C) (1.4 g) was added. The mixture is hydrogenated until quantitative hydrogen uptake. The catalyst is filtered off with suction, washed well with ethanol and the filtrate is evaporated to dryness in vacuo. Yield: 12.80 g (90.1%) of the title compound as a yellowish highly viscous oil.

analysis:

Calculated: C 39.72, H 4.89, F 22.73, N 7.88%, S 2.26; Found: C 39.72, H 4.87, F 22.77, N 7.90, S 2.24.

c) 1- {3-Oxapentane-1,5-dicarboxylic acid-1-oyl-5- [1- (4-perfluorooctylsulfonyl) piperazine] amide} -7- [2,6-N, N'-bis- ( 1-Oa-D-carbonylmethyl-manopyranose)] - L-lysyl-4,10-bis- [1,4,7-tris (carboxymethylmethyl) -10- (3-aza-4-oxo-5-methyl-5) -yl-pentanoyl)] - 1,4,7,10-tetraazacyclododecane, digadolinium complex

5.54 g (8.8 mmol; 2.2 mol equivalents based on the amine component of Example 24b) gadolinium complex of 10- (4-carboxy-1-methyl-2-oxo-3-azabutyl) -1,4, 7, 10-tetraazacyclododecane-1,4,7-triacetic acid (described in patent application DE 197 28 954 Cl in Example 31h) and 0.37 g (8.8 mmol) of anhydrous lithium chloride are dissolved with stirring at 40 ° C in A total of 1.01 g (8.8 mmol) of N-hydroxysuccinimide and 5.68 g (4.0 mmol) of the title compound of Example 24b in 40 ml of absolute dimethylsulfoxide were added at 60 ° C. After cooling to room temperature, the reaction is quenched

To the solution was added 1.82 g (8.8 mmol) of N, N'-dicyclohexylcarbodiimide and stirred at room temperature for 12 hours. Sufficient acetone is added to the resulting suspension to completely precipitate the title compound, the precipitate is filtered off with suction, dried, mixed with water, the insoluble dicyclohexylurea is filtered off and the filtrate is filtered through an Amicon® YM-3 ultrafiltration membrane (eur off: 3000 Da). to desalinate and remove the low molecular weight components. The retentate is then lyophilized. 8.52 g (80.6% based on the charged diamine component) of the product are obtained in the form of a colorless lyophilisate containing 6.09% water (Karl-Fischer).

Analysis (calculated on the anhydrous substance):

Calculated: C 38.61, H 4.76, N 9.53, F 12.21, Gd 11.89, S 1.12;

Found: C 38.57, H 4.82, N 9.52, F 12.21, C-d 11.93, 1.15%

WITH.

Example 25

a) 1,7-Bis (benzyloxycarbonyl) -4- (3-oxapentane-1,5-dicarboxylic acid-1-oyl-5- [1- (4-perfluorooctylsulfonyl) piperazine] amrd) -10-2, 6- N, N'-bis (1-OaD- (5-carbonyl) pentyl-2,3,4,6-tetra-O-benzylmanopyranose)) - L-lysyl-1,4,7,10-tetraazacyclododecane

To a solution of 27.0 g (24.4 mmol) of the secondary amine prepared in Example 15a in a mixture of 150 mL of tetrahydrofuran and 15 mL of chloroform at 0 ° C under a nitrogen atmosphere was added 35, 80 g (25.0 mmol) of the title compound. of Example 17e in 250 ml of tetrahydrofuran. A total of 18.0 g (36.6 mmol) of EEDQ (2-ethoxy-1-ethoxycarbonyl-1,2-dihydroquinoline) was then added in portions at 0 ° C and the mixture was stirred overnight at room temperature. After evaporation in vacuo to dryness, the oily residue obtained is chromatographed on silica gel with n-hexane-isopropanol (20: 1). This gives 49.48 g (80.4% based on the charged secondary amine) of the title compound 103 as a colorless oil.

analysis:

Calculated: C 60.47, H 5.79, F 12.80, N 4.44, S 1.27; Found: C 60.52, H 5.77, F 12.77, N 4.50, S 1.30.

b) 1- {3-Oxapentane-1,5-dicarboxylic acid-1-oyl-5- [1- (4-perfluorooctylsulfonyl) piperazine] amide} -7- [2,6-N, N'-bis- ( 1-0-d- (5-carbonyl) -pentyl-mannopyranose)] - L-lysyl-1,4,7,10- tetraazacyclododecane

25.2 g (10.0 mmol) of the title compound of Example 25a are dissolved in 250 ml of ethanol and 1.8 g of Pearlman catalyst (20% Pd / C) are added. The mixture is hydrogenated until quantitative hydrogen uptake is achieved. The catalyst is filtered off with suction, washed well with ethanol and the filtrate is evaporated to dryness in vacuo. 14.11 g (92.5%) of the title compound are obtained as a yellowish highly viscous oil.

analysis:

Calculated: C 49.60, H 7.20, F 21.17, N 7.34, S 2.10; Found: C 49.62, H 7.17, F 21.20, N 7.30, S 2.14.

c) 1- {3-Oxapentane-1,5-dicarboxylic acid-1-oyl-5- [1- (4-perfluorooctylsulfonyl) piperazine] amide} -7- [2,6-N, N'-bis (1) -O-α- (5-carbonyl) pentyl-manopyranose)] - L-lysyl-4,10-bis- [1,4,7-tris (carboxylatomethyl) -10- (3-aza-4-oxo-5) -methyl-5-yl-pentanoyl) -1,4

7,10-tetraazacyclododecane, digadolinium complex

5.54 g (8.8 mmol; 2.2 mol equivalents based on the amine component of Example 25b) gadolinium complex of 10- (4-carboxy-1-methyl-2-oxo-3-azabutyl) -1,4, 7,10-tetraazacyclododecane-1,4,7-triacetic acid (described in patent application DE 197 28 954 Cl in Example 31h) and 0.37 g (8.8 mmol) of anhydrous lithium chloride are dissolved at 40 ° C under stirring 60 ml of absolute dimethylsulphoxide are added in total at this temperature

104

1.01 g (8.8 mmol) of N-hydroxysuccinimide and 6.10 g (4.0 mmol) of the title compound of Example 25b in 40 ml of absolute dimethylsulfoxide. After cooling to room temperature, 1.82 g (8.8 mmol) of N, N'-dicyclohexylcarbodiimide was added to the reaction solution and stirred at room temperature for 12 hours. Sufficient acetone is added to the resulting suspension to completely eliminate the title compound, the precipitate is filtered off with suction, dried, mixed with water, the insoluble dicyclohexylurea is filtered off and the filtrate is filtered through an Amicon® YM-3 ultrafiltration membrane (cut off: 3000 Da). ) to dewater and remove the low molecular weight components. The retentate is then lyophilized. 9.26 g (84.0% based on the charged diamine component) of the product are obtained in the form of a colorless lyophilisate which contains 5.89% of water (Karl-Fischer).

Analysis (calculated on the anhydrous substance):

Calculated: C 40.52, H 5.16, N 9.15, F 11.72, Gd 11.41, S 1.16;

Found: C 40.57, H 5.20, N 9.12, F 11.69, Gd 11.43, 1.18%

WITH.

Example 26

a) 6-N-tert-butyloxycarbonyl-2-N-benzyloxycarbonyl-L-lysine- [1- (4-perfluorooctylsulfonyl) piperazine] amide

19.02 g (50.0 mmol) of α-N- (benzyloxycarbonyl) -ε-Ν - (tert-butyloxycarbonyl) -L-lysine (commercially available from Bachem) are dissolved in 150 ml of absolute tetrahydrofuran. At 0 ° C, 8.31 g (50.0 mmol) of carbonyldiimidazole and 5.03 g (50.0 mmol) of triethylamine in 75 ml of dry tetrahydrofuran are added dropwise and the mixture is stirred at this temperature for 10 minutes. A solution of 48.42 g (50.0 mmol) of perfluorooctylsulfonylpiperazine and 5.03 g (50.0 mmol) of triethylamine in 250 ml of dry tetrahydrofuran is then added dropwise at 0 ° C. The mixture was stirred overnight, the tetrahydrofuran was evaporated in vacuo

105 and the remaining oil was chromatographed on silica gel with n-hexane-isopropanol (15: 1). Yield: 49.48 g (80.4% based on charged secondary amine) of the title compound as a colorless oil.

analysis:

H, 3.79; N, 6.02; F, 34.70; S, 3.45;

Found: C 40%, H 3.82, N 6.02, F 34.67, S 3.48.

b) 6-N-tert-butyloxycarbonyl-L-lysine- [1- (4-perfluorooctylsulfonyl) piperazine] amide

30.0 g (32.2 mmol) of the title compound of Example 26a was dissolved in 300 ml of isopropanol and 1.5 g of Fearlman catalyst (20% Pd / C) was added. The mixture was hydrogenated for 10 hours at room temperature. After this time, thin-layer analysis showed that the hydrogenolytic cleavage of the benzyloxycarbonyl protecting group proceeded quantitatively. The catalyst is filtered off with suction, the filtrate is evaporated to dryness in vacuo and the residue is chromatographed on silica gel. Yield: 25.13 g (98.0%) of the title compound as a colorless oil.

analysis:

Calculated: C 34.68, H 3.67, F 40.55, N 7.03, S 4.03; Found: C, 34.72; H, 3.70; F, 40.60; N, 7.01; S, 3.98.

c) 6-N-tert-butyloxycarbonyl-2-- [1-SaD- (2-carbonyl) ethyl-2,3,4,6-tetra-O-acetylmanopyranose] -L-lysine- [1- (4) (perfluorooctylsulfonyl) piperazine] amide

Dissolve 15.53 g (35.60 mmol) of 3- (2,3,4,6-tetra-O-acetyl-1-thio-α-D-manopyranosyl) propionic acid (prepared according to: J.) in 300 mL of dry tetrahydrofuran. Haensler et al., Bioconjugate Chem., 4, 85 (1993), Chipowsky, S. and Lee, YC:

Synthesis of 1-thioaldosides; Carbohydrate Research 31, 339-346) and 3.60 g (35.60 mmol) of triethylamine. The solution was cooled to -15 ° C

106 DEG -20 DEG C. and a solution of isobutyl chloroformate in 75 ml of dry tetrahydrofuran is slowly added dropwise with stirring at this temperature, the addition rate being chosen such that the internal temperature does not exceed -10 DEG C. After 15 min at -15 ° C, a solution of the title compound of Example 22b (28.35 g, 35.60 mmol) and ziethylamine (3.60 g, 35.60 mmol) in dry tetrahydrofuran (200 mL) was added dropwise at 20 ° C. After one hour at -15 ° C and two hours at room temperature, the reaction solution is evaporated to dryness in vacuo. The residue is taken up in 250 ml of ethyl acetate and washed twice with 100 ml of saturated sodium bicarbonate solution and once with 200 ml of water. After drying the organic phase over sodium sulfate, the solution is filtered and the ethyl acetate is evaporated in vacuo. The oily residue was then purified by chromatography on silica gel with ethyl acetate-n-hexane (1:25). 34.21 g (79.1%) of the title compound are obtained as a colorless, very viscous oil.

analysis:

Calculated: C 39.54, H 4.23, N 4.61, F 26.58, S 5.28; Found: C 39.49, H 4.21, N 4.59, F 26.52, S 5.31.

d) 6-N-tert-butyloxycarbonyl-2- [1- [S-α-D- (2-carbonyl) ethyl] mannopyranose [-L-lysine- [1- (4-perfluorooctylsulfonyl) piperazine] amide

The resulting compound of Example 26c (29.93 g, 24.64 mmol) was suspended in 400 mL of absolute methanol and a catalytic amount of sodium methoxide was added at 5 ° C. After 3 hours, TLC (9: 1 chloroform eluent) showed the reaction to proceed quantitatively. The resulting clear reaction solution was neutralized by addition of Amberlite ® IR 120 (H ⁺ form), the ion exchanger was filtered off with suction and the methanolic filtrate thus obtained was evaporated to dryness in vacuo. The resulting amorphous residue was purified by silica gel chromatography (2-propanoiethyl acetate-n-hexane, 1: 1: 15) to give 23.42 g (90.8%) of the product as a colorless viscous oil.

107

analysis:

Calculated: C 36.72, H 4.14, N 5.35, F 30.85, S 6.13;

Found: C, 36.69; H, 4.11; N, 5.35; F, 30.82; S, 6.11.

e) 2-N- [1-S-α-D- (2-carbonyl) ethyl-mannopyranose] -L-lysine- [1- (4-perfluorooctylsulfonyl) piperazine] amide

20.93 g (20.0 mmol) of the title compound of Example 26d are dissolved at 0 ° C with vigorous stirring in a mixture of 50 ml of trifluoroacetic acid and 100 ml of dichloromethane and the solution is stirred at this temperature for 10 minutes. It is then evaporated to dryness in vacuo and the residue is taken up in 150 ml of water. The aqueous product solution thus obtained is adjusted to pH 9.5 by dropwise addition of 2 molar aqueous sodium hydroxide solution. The product solution is filtered through an Amicon® YM-3 ultrafiltration membrane (cut off: 3000 Da) to filter off and remove any low molecular components present. The retentate is then lyophilized. 17.79 g (94.2%) of the free amine is obtained in the form of a colorless lyophilisate containing 3.09% water (Karl-Fischer).

Analysis (calculated on the anhydrous substance):

Calculated: C 34.26, H 3.73, N 5.92, F 34.12, S 6.77;

Found: C 34.26, H 3.79, N 5.88, F 34.07, S 6.80.

f) 2-N- [1-SaD- (2-carbonyl) ethyl-mannopyranose] -6-N- [1,4,7-tris (carboxylatomethyl) -10- (3-aza-4-oxo-5- methyl-5-yl-pentancyl) -1,4,7-10-tetraazacyclododecane] -L-lysine- [1- (4-perfluorooctylsulfonyl) piperazine] amide, gadolinium complex

5.54 g (8.8 mmol; 2.2 mol equivalents, based on the amine component of Example 26e) of the gadolinium complex of 10- (4-carboxy-1-methyl-2-oxo-3-azabutyl) -1,4 7, 10-tetraazacyclododecane-1,4,7-triacetic acid (described in patent application DE 197 28 954 Cl in Example 31h) and 0.37 g (8.8 mmol) of anhydrous lithium chloride are dissolved at 40 ° C with stirring. in 60 ml of absolute dimethylsulfoxide and added at this temperature totally

108

1.01 g (8.8 mmol) of N-hydroxysuccinimide and 3.78 g (4.0 mmol) of the title compound of Example 26e in 40 ml of absolute dimethylsulfoxide. After cooling to room temperature, 1.82 g (8.8 mmol) of N, N'-dicyclohexylcarbodiimide was added to the reaction solution and stirred at room temperature for 12 hours. Sufficient acetone is added to the resulting suspension to completely precipitate the title compound, the precipitate is filtered off with suction, mixed with water, the insoluble dicyclohexylurea is filtered off, and the filtrate is filtered through an Amicon® YM-3 ultrafiltration membrane (cut off: 3000 Da). desalted and the low molecular weight components were removed. The retentate is lyophilized. Yield 5.17 g (83.0%) of a colorless lyophilisate containing 4.43% water (Karl-Fischer).

Analysis (calculated on the anhydrous substance):

Calculated: C 35.45, H 4.07, N 8.09, F 20.72, Gd 10.09, S 4.11;

Found: C 35.50, H 4.01, N 8.12, F 20.68, Gd 10.13, 4.14%

WITH.

Example 27

a) 6-N-Benzyloxycarbonyl-2-N- (1-O-β-D-carbonylmethyl-2,3,4,6-tetra-O-benzylglucopyranose) -L-lysine- [1- (4-perfluorooctylsulfonyl) piperazine] amide

8.02 g (13.4 mmol) of 1-carboxymethyloxy-2,3,4,6-tetra-O-benzyl-PB-glucopyranoside (described in patent application DE 197 28 954 C1 in Example 46a) and 3.24 g (28.14 mmol) of N-hydroxysuccinimide was dissolved in 100 ml of dimethylformamide and a total of 5.80 g (28.14 mmol) of N, N'-dicyclohexylcarbodiimide was added portionwise at 0 ° C. The mixture was then stirred at this temperature for a further 3 hours. To the thus obtained active ester solution was added dropwise a solution of 11.13 g (13.4 mmol) of the title compound of Example 1c in 50 ml of DMF, pre-cooled to 0 ° C. The mixture was stirred at 0 ° C for 2 hours and then at room temperature for 12 hours

109 temperature. The precipitated dicyclohexylurea is filtered off and the solvent is evaporated to dryness. The residue is chromatographed on silica gel with dichloromethane-ethanol (20: 1), using a solvent gradient with a continuous increase in the proportion of ethanol in the mixture. Yield: 12.67 g (67.0% of theory) of the title compound as a colorless, very viscous oil.

analysis:

Calculated: C 52.77, H 4.50, N 3.97, F 22.89, S 2.27; Found:% C, 52.75;% H, 4.61;% N, 3.98;% F, 22.94;% S, 2.26%.

b) 2-N- (Ι-Ο-β-D-carbonylmethyl-glucopyranose) -L-lysine- [1- (4-perfluorooctylsulfonyl) piperazine] amide

11.52 g (8.17 mmol) of the compound prepared in Example 27a are dissolved in 100 ml of ethanol and 0.5 g of Pearlman catalyst (20% Pd / C) is added. The mixture is subjected to hydrogenation at room temperature and a hydrogen pressure of 1 atm until there is no more hydrogen uptake. Then, the catalyst is filtered off, washed thoroughly with ethanol (3 x about 40 ml) and the filtrate is evaporated to dryness in vacuo. Yield: 7.36 g (98.4%) of the title compound as a very viscous, colorless oil.

analysis:

Calculated: C 34.07, H 3.63, N 6.11, F 35.24, S 3.50; Found: C, 34.11; H, 3.59; N, 6.08; F, 35.23; S, 3.52.

c) 2-N- (1-O-β-D-2-Carbonylmethyl-glucopvranose) -6-N- [1,4,7-tris (carboxylatomethyl) -10- (3-aza-4-oxo-5-methyl) -5-yl-pentanoyl) -1,4,7,10-tetraazacyclododecane] -L-lysine- [1- (4-perfluorooctylsulfonyl) piperazine] amide, gadolinium complex

9.98 g (15.84 mmol; 2.2 mol equivalents based on the amine component of Example 27b) gadolinium complex of 10- (4-carboxy-1-methyl-2-oxo-3-azabutyl) -1,4 , 7,10-tetraazacyclododecane-1,4,7-triacetic acid (described in patent application

110

DE 197 28 954 Cl in Example 31h) and 0.67 g (15.84 mmol) of anhydrous lithium chloride are dissolved with stirring at 40 ° C in 80 ml of absolute dimethylsulfoxide and a total of 1.82 g (15, 84 mmol) of N-hydroxysuccinimide and 7.25 g (7.19 mmol) of the title compound of Example 27b in 30 ml of absolute dimethylsulfoxide. After cooling to room temperature, N, N'-dicyclohexylcarbodiimide (3.27 g, 15.84 mmol) was added to the reaction solution and stirred at room temperature for 12 hours. Sufficient acetone is added to the resulting suspension to precipitate the final compound completely, the precipitate is filtered off with suction, mixed with water, the insoluble dicyclohexylurea is filtered off and the filtrate is filtered through an Amicon® YM-3 ultrafiltration membrane (cut off: 3000 Da). ) to desalinate and remove the low molecular weight components. The retentate is then lyophilized. 9.11 g (83.0%) of product are obtained in the form of a colorless lyophilisate which contains 4.02% of water (Karl-Fischer).

Analysis (calculated on the anhydrous substance):

Calculated: C 35.37, H 4.02, N 8.25, F 21.13, S 2.10, Gd 10.29;

Found:% C, 35.42;% H, 7;% N, 8.18;% F, 21.09;% S, 2.06;

Example 28

a) 2-N-Trifluoroacetyl-1'-lysine- [1- (4-perfluorooctylsulfonyl) piperazine] amide

10.0 g (11.46 mmol) of the compound prepared in Example 1b were dissolved in 100 ml of ethanol and 1.0 g of Pearlman catalyst (20% Pd / C) was added. The mixture is subjected to hydrogenation until quantitative consumption of hydrogen. Then the catalyst is filtered off, washed with ethanol and the filtrate is evaporated to dryness in vacuo. Yield: 8.85 g (97.5% of theory) of the title compound as a viscous colorless oil.

111

analysis:

Calculated: C 30.31, H 2.54, N 7.07, F 47.95, S 4.05; Found: C 30.36, H 2.50, N 7.11, F 47.99, S 4.00.

b) 2-N-Trifluoroacetyl-6-N- [1-OaD- (5-carbonyl) pentyl-2,3,4,6-tetra-O-benzyl-mannopyranose] -L-lysine- [1- (4) -perfluóroktylsulfonyl) -piperazine] amide

To a solution of 29.0 g (36.6 mmol) of the title compound of Example 28a and 4.05 g (40.26 mmol) of triethylamine in 100 mL of DMF cooled to 0 ° C was added dropwise a solution of 27.51 g (36.6 mmol). mmol) of the title compound of Example 17c in 150 mL of dimethylformamide. After the addition was complete, the mixture was stirred at 0 ° C for one hour and then at room temperature overnight. It is then evaporated to dryness in vacuo and the residue is taken up in 300 ml of ethyl acetate. After insoluble matter is filtered off, the filtrate is washed twice with 10 ml of 5% aqueous soda solution. The organic phase is dried over magnesium sulphate and evaporated to dryness in vacuo, chromatography on silica gel (25: 1). Thus, 42.05 c (80.40%) of the title compound are obtained as a colorless oil.

The residue is treated with n-hexane-isocropanol

analysis:

Calculated: C 50.42, H 4.51, N 7.96, F 26.59, S 2.24; Found:% C, 50.38;% H, 4.50;% N, 7.91;% F, 26.62;

c) 6-N- [1-O-α-D- (5-carbonyl) pentyl-2,3,4,6-tetra-O-benzyl-manopyranose] -L-lysine- [1- (4-perfluorooctylsulfonyl) piperazine] amide

20.0 g (14.0 mmol) of the compound prepared in Example 28b were dissolved in 150 ml of methanol. To this solution was added a solution of 2.8 g (70.0 mmol) of sodium hydroxide in 25 mL of distilled water and stirred at 50 ° C for 0.5 h. According to thin layer chromatography, the cleavage of the protecting groups is quantitative after this time. After evaporation to dryness in vacuo, the residue is traces of water by repeated codistillation with ethanol. The residue is subjected to

112 chromatography on n-hexane-isopropanol (20: 1). Yield: 16.66 g (89.3% of theory) of the title compound as a colorless oil.

analysis:

Calculated: C 52.25, H 4.91, N 4.20, F 24.22, S 2.41; Found:% C, 52.30;% H, 4.90;% N, 4.18;% F, 2.32%.

C; 6-N- [1-O-α-D- (5-Carbonyl) pentyl-manopyranose] -L-lysine- [1- (4-perfluorooctylsulfonyl) piperazine] amide

15.0 g (11.25 mmol) of the compound prepared in Example 28c were dissolved in 150 ml of a 10: 1 mixture of ethanol and water and 1.0 g of Pearlman's catalyst (20% Pd / C) was added. The mixture is subjected to hydrogenation at room temperature and a pressure of 1 atm until quantitative consumption of hydrogen. The catalyst was then filtered off, washed with ethanol: water (10: 1) and the filtrate was evaporated to dryness in vacuo. This gave 10.77 g (98.4%) of the title compound as a viscous colorless oil.

analysis:

Calculated: C 37.04, H 4.25, N 5.76, F 33.20, S 3.30; Found: C 37.06, H 4.20, N 5.81, F 33.19, S 3.30.

e '6-N- [1-O-α- (5-Carbonyl) pentyl-manopyranose] -2-N- [1,4,7-tris (carboxylatomethyl) -10- (3-aza-4-oxo-) 5-methyl-5-yl-pentanoyl) -1,4,7,10-tetraazacyclododecane] -L-lysine- [1- (4-perfluorooctylsulfenyl) piperazine] amide, gadolinium complex

5.54 g (8.8 mmol; 2.2 mol equivalents, based on the amine component of Example 28d) of the gadolinium complex of 10- (4-carboxy-1-methyl-2-oxo-3-azabutyl) -1,4 7,10-tetraazacyclododecane-1,4,7-triacetic acid (described in patent application DE 197 28 954 C1 in Example 31h) and 0.37 g (8.8 mmol) of anhydrous lithium chloride are dissolved at 40 ° C with stirring. in 60 ml of absolute dimethylsulfoxide and added at this temperature totally

113

1.01 g (8.8 mmol) of N-hydroxysuccinimide and 3.89 g (4.0 mmol) of the title compound of Example 28d in 60 ml of absolute dimethylsulfoxide. After cooling to room temperature, 1.82 g (8.8 mmol) of N, N'-dicyclohexylcarbodiimide was added to the reaction solution and stirred for 12 hours at room temperature. Sufficient acetone is then added to the resulting suspension to completely precipitate the title compound, the precipitate is filtered off with suction, mixed with water, the insoluble dicyclohexylurea is filtered off and the filtrate is filtered through an Amicon® YM-3 ultrafiltration membrane (cut off: 3000 Da), to desalinate and remove the low molecular weight components. The retentate is then lyophilized. 4.81 g (75.9%) of product are obtained in the form of a colorless lyophilisate which contains 8.98% of water (Kari-Fischer).

Analysis (calculated on the anhydrous substance):

Calculated: C 37.15, H 4.39, N 7.96, F 20.38, Gd 9.92, S 2.02;

Found: C 37.27, H 4.40, N 8.02, F 20.31, Gd 10.00, 1.98%

WITH.

Example 29

a) 1,7-Bis (benzyloxycarbonyl) -4- (1-O-β-D-carbonylmethyl-2,3,4,6-tetra-O-benzyl-galactopyranose) -10- (3-oxapentane-1,5- -1,4,7,10-tetraazacyclododecane dicarboxylic acid-1-oyl-5- [1- (4-perfluorooctylsulfonyl) piperazine] amide}

To a solution of 27.0 g (24.4 mmol) of the secondary amine prepared in Example 15a in a mixture of 150 mL of tetrahydrofuran and 15 mL of chloroform at 0 ° C under nitrogen was added a solution of 35.80 g (25.0 mmol) of the resulting amine. of the compound of Example 17e in 250 mL of tetrahydrofuran. A total of 18.0 g (36.6 mmol) of EEDQ (2-ethoxy-1-ethoxycarbonyl-1,2-6-hydroquinoline) was added portionwise at 0 ° C and stirred overnight at room temperature. After evaporation in vacuo to dryness, the residual oil is chromatographed on silica gel

114 in n-hexane-isopropanol (20: 1). 32.11 g (73.0% of the charged secondary amine) of the title compound are obtained as a colorless oil.

analysis:

Calculated: C 54.09, H 4.72, F 19.14, N 4.98, S 1.90; Found: C, 54.12, H, 4.77, F, 19.17, N, 5.03, S, 1.90.

b) 1- (Ι-Ο-β-D-Carbonylmethyl-galactopyranose) -7- (3-oxapentane-1,5-dicarboxylic acid-1-oyl-5- [1- (4-perfluorooctylsulfonyl) piperazine] amide) 1,4,7,10-tetraazacyclododecane

30.0 g (17.77 mmol) of the compound prepared in Example 29a are dissolved in 250 ml of ethanol and 3.0 g of Pearlman catalyst (20% Pd / C) are added. The mixture is subjected to hydrogenation until a quantitative amount of hydrogen is absorbed. The catalyst is then filtered off, washed thoroughly with ethanol and the filtrate is evaporated to dryness in vacuo. 17.89 g (95.1%) of the title compound are obtained as a highly viscous yellowish oil.

analysis:

Calculated: C 36.30, H 4.09, F 30.50, N 7.94, S 3.03; Found: C 36.26, H 4.12, F 30.46, N 7.90, 3.04% S.

c) 1- (Ι-Ο-β-D-Carbonylmethyl-galactopyranose) -7- {3-oxapentane-1,5-dicarboxylic acid-1-oyl-5- [1- (4-perfluorooctylsulfonyl) piperazine] amide] -4,10-bis [1,4,7-tris (carboxylatomethyl) -10- (3-aza-4-oxo-5-methyl-5-ylpentanoyl) -1,4,7,10-tetraazacyclododecane] -1,4,7,10-tetraazacyclododecane, digadolinium complex

5.54 g (8.8 mmol; 4.4 mol equivalents, based on the amine component of Example 29b) gadolinium complex 10- (4-carboxy-1-methyl-2-oxo-3-azabutyl) -1,4 7,10-tetraazacyclododecane-1,4,7-triacetic acid (described in patent application DE 197 28 954 C1 in Example 31h) and 0.37 g (8.8 mmol) of anhydrous lithium chloride are dissolved at 40 ° C with stirring. A total of 1.01 g (8.8 mmol) of N-hydroxysuccinimide and 2.11 g (2.0 mmol) of the title compound of Example 29b in 25 ml of absolute dimethylsulfoxide are added at 60 DEG C. in 60 ml of absolute dimethyl sulfoxide. After cooling to room temperature, 1.82 g (8.8 mmol) of N, N'-dicyclohexylcarbodiimide was added to the reaction solution and stirred at room temperature for 12 hours. Sufficient acetone is added to the resulting suspension to completely precipitate the resulting compound, the precipitate is filtered off with suction, mixed with water, the insoluble dicyclohexylurea is filtered off, and the filtrate is filtered through an Amicon® YM-3 ultrafiltration membrane (cut off: 3000 Da). desalted and the low molecular weight components were removed. The retentate is lyophilized. Yield 3.29 g (72.2% based on the charged amine component) of a colorless lyophilisate containing 5.99% water (Karl-Fischer).

Analysis (calculated on the anhydrous substance):

Calculated: C 36.84, H 4.37, N 9.82, F 14.15, Gd 19.63, S 1.40;

Found: C 36.87, H 4.40, N 9.82, F 14.09, Gd 19.59, 1.38%

WITH.

Example 30

a) 3- (1-O-α-D-2,3,4,6-Tetra-O-benzylmanopyranose) -2-N-benzyloxycarbonyl-L-serine methyl ester

21.42 g (39.61 mmol) of 2,3,4,6-tetra-O-benzyl-α-D-mancpyranose (prepared according to: F. Kong et al., J. Carbohydr. Chem., 16; 6; 1997) 877-890) is dissolved in 500 ml of dry acetonitrile. After cooling the solution to 5 ° C, a solution of 13.23 g (59.52 mmol) of trifluoromethanesulfonic acid trimethylsilyl ester in 30 ml of acetonitrile is slowly added dropwise with stirring at this temperature, followed by a solution of 20.06 g (79.21 mmol) of N-benzyloxycarbonyl- Of L-serine methyl ester (commercially available from Bachem) in 50 ml of acetonitrile, the rate of addition being selected such that the internal temperature does not exceed 116 and 10 ° C. After 15 hours at room temperature, the reaction solution is evaporated to dryness in vacuo. The residue is taken up in 250 ml of ethyl acetate, washed twice with 100 ml of saturated sodium bicarbonate solution and once with 200 ml of water. After drying the organic phase over sodium sulfate and filtration, the ethyl acetate is evaporated in vacuo and oils, and the residue is chromatographed on silica gel with ethyl acetate-n-hexane (1: 5). Yield: 23.60 g (76.8%) of the title compound as a colorless oil.

analysis:

H, 6.37; N, 1.81.

Found:% C, 71.19;% H, 6.41;% N, 1.79.

b) 3- (1-O-α-D-2,3,4, β-Tetra-O-benzyl-manopyranose) -2-N-benzyloxycarbonyl-L-serine

10.0 g (12.90 mmol) of the compound prepared in Example 30a are dissolved in a mixture of 20 ml of methanol, 20 ml of water and 50 ml of tetrahydrofuran. Thereafter, 0.47 g (19.35 mmol) of lithium hydroxide dissolved in 25 ml of distilled water is added at room temperature and the mixture is stirred for 6 hours at 60 ° C. Following a thin layer chromatography control (eluent: methylene chloride-methanol 10: 1), the methyl ester of Example 30a is completely saponified after this time. The reaction solution is evaporated to dryness in vacuo and the residue is taken up in 250 ml of ethyl acetate while hot (60 ° C). The ethyl acetate phase thus obtained is washed twice with 50 ml of 15% aqueous hydrochloric acid and then once with 100 ml of distilled water. The organic phase is dried over magnesium sulphate, filtered and the solvent is evaporated to dryness in vacuo. The residue was chromatographed on silica gel with n-hexane-ethyl acetate (5: 1). Yield: 8.40 g (85.7%) of the title compound as a colorless oil.

analysis:

H, 6.22; N, 1.84.

Found:% C, 70.97;% H, 6.30;% N, 1.78.

117

c) 3- (1-O-α-D-2,3,4,6-Tetra-O-benzyl-manopyranose) -2-N-benzyloxycarbonyl-L-serine- [1- (4-perfluorooctylsulfonyl) piperazine] amide

To a solution of 13.86 g (24.40 mmol) of 1-perfluoro-octylsulfonylpiperazine (prepared according to DE 19603033) in a mixture of 150 ml of tetrahydrofuran and 15 ml of chloroform is added dropwise at 0 ° C under nitrogen atmosphere 20.57 g (27.0 mmol). of the acid prepared in Example 30b dissolved in 50 ml of tetrahydrofuran. A total of 18.0 g (36.60 mmol) of EEDQ (2-methoxy-1-ethoxycarbonyl-1,2-dihydroquinoline) was added portionwise at 0 ° C and the mixture was stirred overnight at room temperature. After evaporation in vacuo, the highly viscous residue is chromatographed on silica gel with n-hexane-isopropanol (15: 1). Yield: 17.0 g (79.6% based on the primary amine used) of the title compound as a colorless viscous oil.

analysis:

Calculated: C 51.53, H 4.23, N 3.15, F 25.65, S 2.41; Found: C 51.48, H 4.27, N 3.10, F 25.71, S 2.35.

d) 3- (1-O-α-D-mannopyranose) -L-serine- [1- (4-perfluorooctylsulfonyl) piperazine] amide

To a solution of 15.0 g (11.41 mmol) of the compound prepared in Example 30c in 200 mL of ethanol was added 1.5 g of Pearlman catalyst (20% Pd / C) and hydrogenated at room temperature and 1 atm to the hydrogen consumption stops (about 8 hours). The catalyst is filtered off with suction, washed thoroughly with ethanol (2 * 100 ml) and the ethanolic filtrate is evaporated to dryness in vacuo. It will be obtained

8.79 g (94.0%) of the title compound as a very viscous colorless oil.

analysis:

Calculated: C 30.78, H 3.20, N 5.13, F 39.41, S 3.91; Found: C, 30.87; H, 3.14; N, 5.19; F, 39.50.

118

e) 3- (Ι-Ο-α-D-Manopyranose) -2-N- [1,4,7-tris (carboxylatomethyl) -10- (3-aza-4-oxo-5-methyl-5-yl) -pentanoyl) -1,4,7,10-tetraazacyclododecane [-L-serine- [1- (4-perfluorooctylsulfonyl) piperazine] amide, gadolinium complex

To a stirred suspension of 5.7 g [9.06 mmol; corresponds to 1.5 mol equivalents of the title compound (primary amine) of Example 30d] cadolinium complex of 10- (4-carbonyloxy-1-methyl-2-oxo-3-azabutyl-1,4,7,10-tetraazacyclododecane-1), Of 4,7-triacetoacetic acid (described in patent application DE 197 28 954 Cl in Example 31h) in 75 ml of absolute dimethylsulfoxide at 70 ° C is added 0.68 g (15.9 mmol) of lithium chloride, after stirring for 30 minutes at 70 ° C. a total of 1.83 g (15.9 mmol) of N-hydroxysuccinimide was added portionwise to the resulting clear reaction solution, and the reaction mixture was maintained at this temperature for 1 hour. 85 mmol) of dicyclohexylcarbodiimide and the reaction solution is stirred for an additional hour at 0 [deg.] C. and then for 12 hours at 22 [deg.] C. To the gadolinium acid N-hydroxysuccinimide ester solution thus obtained.

10- (4-carboxy-1-methyl-2-oxo-3-azabutyl) -1,4,7,10-tetraazacyclododecane-1,4,7-triacetic acid is added dropwise at 22 ° C a solution of 4.94 g (6 , 03 mmol) of the title compound of Example 30d in 15 mL of absolute dimethylsulfoxide and stirred for an additional 12 hours at room temperature. The reaction solution was then slowly added dropwise at 22 ° C to a mixture of 250 ml of acetone and 250 ml of 2-propanol, after 12 hours at 10 ° C the title compound completely precipitated as a yellowish oil. After gently decanting the solvent mixture, the oily product is taken up in 200 ml of distilled water and completely dissolved in the yellowish-colored aqueous solution of the title compound. This solution is first filtered through a membrane filter and then three times ultrafiltration through a YM-3 ultrafiltration membrane (Amicon®, cut off: 3000 Da) to achieve desalination and remove low molecular weight components. The retentate is then lyophilized. There was thus obtained 8.63 g (80.2% based on the title compound)

119 of Example 30d) as a colorless lyophilizate containing 7.65% water.

Analysis (calculated on the anhydrous substance):

Calculated: C 33.57, H 3.80, N 7.83, F 22.57, Gd 10.99, S 2.24;

Found: C 33.57, H 3.76, N 7.82, F 22.63, Gd 11.06, 2.18%

WITH.

Example 31

a) 6-N-Benzyloxycarbonyl-2-N- [Ο-β-D-galactopyranosyl- (1-> 4) glucosyl] -L-lysine- [1- (4-perfluorooctylsulfonyl) piperazine] amide

To a stirred solution of 4.98 g (6.0 mmol) of the title compound of Example 1c in 40 mL of absolute dimethylsulfoxide was added dropwise a solution of 13.3 g (37.2 mmol) of Ο-β-D-galactopyranosyl- (1- 4) -D-glucono-1,5-lactone [lactobionolactor .; prepared according to: a) Williams, T.J. Plessas, N.R. Goldstein, l. J. Carbohydr. Res. 1978, 67, Cl; (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 ml of absolute dimethylsulfoxide. The resulting reaction solution was then stirred at 40 ° C for 14 hours. Absolute 2-propanol (500 ml) was added at room temperature and the colorless precipitate formed was filtered off with suction on frit G4 and rinsed well with a total of 250 ml of absolute 2-propanol. The solid obtained is dissolved in 300 ml of distilled water and filtered through a three-pass filter through a YM-3 ultrafiltration membrane (Amicon®, cut off: 3000 Da). This triple ultrafiltration removes excess lactobionclactone and any low molecular weight components still present. The residue retained in the ultrafiltration membrane is washed completely into 300 ml of distilled water and subjected to lyophilization. Yield 6.51 g (92.7%) of a colorless lyophilisate containing 10.03% water.

120

Analysis (calculated on the anhydrous substance):

Calculated: C 38.98, H 4.05, N 4.79, F 27.58, S 2.74; Found: C, 39.04; H, 4.09; N, 4.82; F, 27.61; S, 2.71.

b) 2-N- [Ο-β-D-Galactopyranosyl- (1-> 4) -gluconosyl] -L-lysine- [1- (4-perfluorooctylsulfonyl ·) piperazine] amide

To a solution of 5.0 g (4.27 mmol) of the compound prepared in Example 31a in 100 mL of ethanol was added 0.5 g of Pearlman catalyst (20% Pd / C) and hydrogenated at room temperature and 1 atm to into quantitative hydrogen consumption. The catalyst is filtered off with suction, washed with ethanol and the filtrate is evaporated to dryness in vacuo. Yield: 4.36 g (98.5%) of the title compound as a viscous colorless oil.

analysis:

Calculated: C 34.76, H 3.99, N 5.40, F 31.51, 3.09% 5; Found:% C, 34.78;% H, 4.04;% N, 5.34.% F, 3.15%.

c) 2-N- [β-D-Galactopyranosyl- (1-> 4) -gluconosyl] -6-N- [1,4,7-tris (carboxylatomethyl) -10- (3-aza-4- oxo-5-methyl-5-yl-peranoyl) -1,4,7,10-tetraazacyclododecane] -L-lysine- [1- (4-perfluorooctylsulfonyl) piperazine] amide, gadolinium complex

5.54 g (8.8 mmol; 2.2 mol equivalents based on the amine component of Example 31b) gadolinium complex of 10- (4-carboxy-1-methyl-2-oxo-3-azabutyl) -1,4 7,10-tetraazaoyl-dododecane-1,4,7-triacetic acid (described in patent application DE 197 28 954 Cl under Example 31h) and 0.37 g (8.8 mmol) of anhydrous lithium chloride are dissolved at 40 ° C with stirring. in 60 ml of absolute dimethylsulfoxide and at this temperature a total of 1.01 g (8.8 mmol) of N-hydroxysuccinimide and 3.85 g (4.0 mmol) of the title compound of Example 31b in 60 ml of absolute dimethylsulfoxide are added. After cooling to room temperature, 1.82 g (8.8 mmol) of N, N'-dicyclohexylcarbodiimide was added to the reaction solution and stirred at room temperature for 12 hours. The resulting suspension

Add sufficient acetone-2-propanol (1: 1) to precipitate the title compound, filter the precipitate by suction, mix with 300 ml of water, filter out the insoluble dicyclohexylurea and filter the filtrate three times through an Amicon® YM- ultrafiltration membrane. 3 (cut off: 3000 Da). This filtration, on the one hand, removes from the desired product the excess gadolinium complex and, on the other hand, any low molecular weight components that are still present. The residue in the ultrafiltration membrane is then completely dissolved in 500 ml of distilled water and subjected to lyophilization. Yield 4.64 g (70.4%) of a colorless lyophilizate containing 10.08% water (Karl-Fischer).

Analysis (calculated on the anhydrous substance):

Calculated: C 35.70, H 4.22, N 7.65, F 19.59, Gd 9.54, S 1.95;

Found: C 35.77, H 4.17, N 7.71, F 19.61, Go 9.60, 1.99%

WITH.

Example 32

a) 6-N-Benzyloxycarbonyl-2-N- (2,3,4,5-pentahydroxy-hexanoyl) -L-lysine- [1- (4-perfluorooctylsulfonyl) piperazine] amide

To a solution of the resulting compound of Example 1c (100.0 g, 120.4 mmol) in dry tetrahydrofuran (500 mL) was added dropwise a solution of 5-gluconolactone (21.45 g, 120.4 mmol) in THF (50 mL) at 50 ° C. 3 hours at 60 ° C and then overnight at room temperature. After evaporation in vacuo to dryness, the residue is chromatographed on silica gel with dichloromethane-ethanol (20: 1). 98.37 g (82%) of a viscous oil are obtained.

analysis:

H, 3.70; F, 32.02; N, 5.55; S, 3.18. Found: C 38.22, H 3.79, F 32.02, N 5.42, S 3.29.

122

b) 2-N- (2,3,4,5-pentahydroxy-hexanoyl) -L-lysine- [1- (4-perfluorooctylsulfonyl) piperazine] amide

To a solution of 100.9 g (100.0 mmol) of the title compound of Example 32a in 2000 mL of ethanol was added 10.0 g of palladium catalyst (10% Pd / C) and the mixture was hydrogenated at room temperature for 12 hours. The catalyst is then filtered off and the filtrate is evaporated to dryness in vacuo. Yield 87.46 g (100%) of the product as a colorless solid.

analysis;

Calculated: C 32.96, H 3.57, N 6.41, S 3.67, F 36.93; Found: C 32.91, H 3.72, N 6.34, S 3.50, F. 36.78.

c) 6-N- [1,4,7-Tris (carboxylatomethyl) -1,4,7,10-retraazacyclododecane-10-β- (pentanoyl-3-aza-4-oxo-5-methyl-5-yl) ]] - 2-N- [1-Oa-D-carbonylmethyl-manopyranose] -L-lysine- [1- (4-perfluorooctylsulfonyl) piperazine] amide, gadolinium complex

50.0 g (54.55 mmol) of the title compound of Example 1e,

6.28 g (54.55 mmol) of N-hydroxysuccinimide, 4.62 g (109.0 mmol) of lithium chloride and 34.35 g (54.55 mmol) of gadolinium complex 1,4,4-tris (carboxylatomethyl) - 10- (Carboxy-3-aza-4-oxo-5-methyl-pent-5-yl) -1,4,7,10-tetraazacyclododecane was dissolved in 400 ml of dimethyl sulfoxide with gentle heating. Add at 10 ° C

16.88 g (81.8 mmol) of N, N-dicyclohexylcarbodiimide and the mixture was stirred overnight at room temperature. The solution was then poured into 3000 ml of acetone and stirred for 10 minutes. The precipitated solid bath was filtered off and purified by chromatography on RP-18 phase in a water-ethanol-acetonitrile (gradient) system. 75.9 g (91.0%) of product are obtained as a colorless solid which contains 8.6% water.

Analysis (calculated on the anhydrous substance):

Calculated: C 35.34, H 4.09, N 8.24, S 2.10, F 21.12,

10.28% Gd;

123

Found: C 35.28, H 4.15, N 8.19, S 2.15, F 21.03, Gd 10.14.

Example 33

a) 6-N-Benzyloxycarbonyl-2-N- (2,3,4,5-pentahydroxy-hexancyl) -L-lysine- [1- (4-perfluorooctylsulfonyl) piperazine] amide

To a solution of the title compound of Example 1c (100.0 g, 120.4 mmol) and triethylamine (12.18 g, 120.4 mmol) in dry tetrahydrofuran (500 mL) was added dropwise a solution of 21.45 g (120.4 mmol) at 50 ° C. 5-gluconolactone in 50 ml tetrahydrofuran. The mixture was stirred at 60 ° C for 3 hours and then at room temperature overnight. 400 ml of 5% aqueous hydrochloric acid are then added, stirred for 5 minutes at room temperature, sodium chloride is added, the organic phase is separated, dried over magnesium sulphate, evaporated to dryness in vacuo and the residue is chromatographed on silica gel. dichloromethane-ethanol (20: 1). 100.97 g (82%) of the product are obtained as a viscous oil.

analysis:

Calculated: C 37.58, H 3.45, F 31.58, N 5.48, S 3.14; Found: C 37.72, H 3.59, F 31.72, N 5.42, S 3.29.

b) 2-N- (2,3,4,5-pentahydroxy-hexanoyl) -L-lysine- [1- (4-perfluorooctylsulfonyl) piperazine] amide

To a solution of 100.9 g (100.0 mmol) of the title compound of Example 32a in 2000 mL of ethanol was added 10.0 g of palladium catalyst (10% Pd / C) and the mixture was hydrogenated at room temperature for 12 hours. The catalyst is then filtered off and the filtrate is evaporated to dryness in vacuo. Yield 87.46 g (100%) of the product as a colorless solid.

analysis:

Calculated: C 32.96, H 3.57, N 6.41, S 3.67, F 36.93;

124

Found: C 32.91, H 3.72, N 6.34, S 3.50, F. 36.78.

c) 6-N- [1,4,7-Tris (carboxylatomethyl) -1,4,7,10-tetraazacyclododecane-10-N- (pentanoyl-3-aza-4-oxo-5-methyl-5-yl) )] - 2-N- [1-O-α-D-carbonylmethyl-maropyranose] -L-lysine- [1- (4-perfluorooctylsulfonyl) piperazine] amide, gadolinium complex

50.0 g (54.55 mmol) of the title compound of Example Ie,

6.28 g (54.55 mmol) of N-hydroxysuccinimide, 4.62 g (109.0 mmol) of lithium chloride and 34.35 g (54.55 mmol) of gadolinium complex

1,4,7-tri (carboxylatomethyl) -10- (carboxy-3-aza-4-oxo-5-methylpent-5-yl) -1,4,7,10-tetraazacyclododecane dissolves with gentle warming in 400 ml of dimethylsulfoxide. Add at 10 ° C

16.88 g (81.8 mmol) of N, N-dicyclohexylcarbodiimide and the mixture was stirred overnight at room temperature. The solution was then poured into 3000 ml of acetone and stirred for 10 minutes. The precipitated solid is filtered off and purified by chromatography on RP-18 phase in a water-ethanol-acetonitrile (gradient) system. 75.9 g (91.0%) of product are obtained as a colorless solid which contains 8.6% water.

Analysis (calculated on the anhydrous substance):

Calculated: C 35.34, H 4.09, N 8.24, S 2.10, F 21.12, Gd 10.28;

Found: C 35.28, H 4.15, N 8.19, S 2.15, F 21.03, Gd 10.14.

Under the conditions of Example 1f, mannose is replaced by glucose or galactose.

Example 34

a) 6-N-Benzyloxycarbonyl-2-N- [1-Oa-D-carbonylmethyl- (2,3,4,6-tetra-O-benzyl-glucopvranose)] - L-lysine- [1- (4-perfluorooctylsulfonyl)] piperazine] -amide

125

To a solution of 100.0 g (120.4 mmol) of the title compound of Example 1c, 72.1 g (120.4 mmol) of 10aD-carboxymethyl-2,3,4,6-tetra-O-benzyl-glucopyranose and 13, 86 g (120.4 mmol) of N-hydroxysuccinimide in 500 ml of dimethylformamide are added at 0 ° C with 41.27 g (200.0 mmol) of N, N-dicyclohexylcarbodiimide. The mixture was stirred at 0 ° C for 3 hours and then at room temperature overnight. The precipitated urea is then filtered off and the filtrate is evaporated to dryness in vacuo. The residue is chromatographed on silica gel with dichloromethane-ethanol (20: 1). 136.1 g (87%) of the product are obtained as a viscous oil.

analysis:

Calculated: C 57.32, H 4.89, N 4.31, F 24.86, S 2.47; Found: C 57.48, H 5.04, N 4.20, F 24.69, S 2.38.

d) 2-N- [1-O-α-D-carbonylmethylglucopyranose] -L-kizine- [1- (4-perfluorooctylsulfonyl) piperazine] amide

To a solution of 130.0 g (100.0 mmol) of the title compound of Example 34a in 2000 mL of ethanol was added 10.0 g of palladium catalyst (10% Pd / C) and hydrogenated at room temperature for 12 hours. The catalyst is then filtered off and the filtrate is evaporated to dryness in vacuo. Yield 91.7 g (100%) of the product as a colorless solid.

analysis:

Calculated: C 34.07, H 3.63, N 6.11, S 3.50, F 35.24; Found: C, 33.92; H, 3.71; N, 6.02; S, 3.42; F, 35.33.

c) 6-N- [1,4,7-Tris (carboxylatomethyl)] - 1,4,7,10-tetraazacyclododecane-10-Ν- (pentanoyl-3-aza-4-oxo-5-methyl-5- yl)] - 2-N- [1-O-α-D-carbonylmethyl-glucopyranose [-L-lysine- [1- (4-perfluorooctyisulfonyl) piperazine] amide, gadolinium complex

50.0 g (54.55 mmol) of the title compound of Example 34b,

6.28 g (54.55 mmol) of N-hydroxysuccinimide, 4.62 g (109.0 mmol)

126 lithium chloride and 34.35 g (54.55 mmol) of the gadolinium complex

1,4,7-tris (carboxylatomethyl) -10- (carboxy-3-aza-4-oxo-5-methylpent-5-yl) -1,4,7,10-tetraazacyclododecane is dissolved in 400 with gentle heating ml of dimethylsulfoxide was added at 10 ° C

16.88 g (81.8 mmol) of N, N-dicyclohexylcarbodiimide and the mixture was stirred overnight at room temperature. The solution was then poured into 3000 ml of acetone and stirred for 10 minutes. The precipitated solid is filtered off and purified by chromatography on RP-18 phase in a water-ethanol-acetonitrile (gradient) system. 75.9 g (91.0%) of product are obtained as a colorless solid which contains 8.6% water.

Analysis (calculated on the anhydrous substance):

Calculated: C 35.34, H 4.09, N 8.24, S 2.10, F 21.12, Gd 10.28;

Found: C 35.26, H 4.18, N 8.14, S 2.158, F 21.01, Gd 10.13.

Example 35

a) 6-N-Benzyloxycarbonyl-2-N- [1-Oa-D-carbonylmethyl- (2,3,4,6-tetra-O-benzyl-galactopyranose)] - L-lysine- [1- (4-perfluorooctylsulfonyl)] piperazine] amide

To a solution of 20.0 g (60.2 mmol) of the title compound of Example 1c, 36.05 g (60.2 mmol) of 10α-D-carboxymethyl-2,3,4,6-tetra-O-oenzyl-galactopyranose and 6α. 93.6 g (60.2 mmol) of N-hydroxysuccinimide in 500 ml of dimethylformamide are added at 0 DEG C. 20.64 g (100.0 mmol) of N, N-dicyclohexylcarbodiimide. The mixture was stirred at 0 ° C for 3 hours and then at room temperature overnight. The precipitated urea is then filtered off and the filtrate is evaporated to dryness in vacuo. The residue is chromatographed on silica gel with dichloromethane-ethanol (20: 1). 68.1 g (87%) of the product are obtained as a viscous oil.

127

Analysis:

Calculated: C 57.32, H 4.89, N 4.31, F 24.86, S 2.47; Found: C 57.47, H 5.05, N 4.19, F 24.72, S 2.29.

b) 2-N- [1-O-α-D-carbonylmethyl-galactopyranose] -L-lysine- [1- (4-perfluorooctylsulfonyl) piperazine] amide

To a solution of the title compound of Example 35a (65.0 g, 50.0 mmol) in ethanol (1000 mL) was added palladium catalyst (10% Pd / C, 5.0 g) and the mixture was hydrogenated at room temperature for 12 hours. The catalyst is then filtered off and the filtrate is evaporated to dryness in vacuo. Yield 45.85 g (100%) of a colorless solid.

analysis:

Calculated: C 34.07, H 3.63, N 6.11, S 3.50, F 35.24; Found: C, 33.93; H, 3.74; N, 6.01; S, 3.39; F, 35.05.

c) 6-N- [1,4,7-Tris (carboxylatomethyl) -1,4,7,10-tetraazacyclododecane-10-β- (pentanoyl-3-aza-4-oxo-5-methyl-5-yl) ]] - 2-N- [1-Oa-D-carbonylmethyl-galactopyranose] -L-lysine- [1- (4-perfluorooctylsulfonyl) piperazine] amide, gadolinium complex

50.0 g (54.55 mmol) of the title compound of Example 35b,

6.28 g (54.55 mmol) of N-hydroxysuccinimide, 4.62 g (109.0 mmol) of lithium chloride and 34.35 g (54.55 mmol) of gadolinium complex

1,4,7-Tris (carboxylatomethyl) -10- (carboxy-3-aza-4-oxo-5-methylpent-5-yl) -1,4,7,10-tetraazacyclododecane dissolves in mild heating 400 ml of dimethylsulfoxide. Add at 10 ° C

16.88 g (81.8 mmol) of N, N-dicyclohexylcarbodiimide and the mixture was stirred overnight at room temperature. The solution was then poured into 3000 ml of acetone and stirred for 10 minutes. The precipitated solid is filtered off and purified by chromatography on RP-18 phase in a water-ethanol-acetonitrile (gradient) system. Yield 37.95 g (91.0%) of a colorless solid containing 8.6% water.

Analysis (calculated on the anhydrous substance):

128

Calculated: 35.34% 10.28% Gd; C, 4.09% H, 8.24% N, 2.10% S, 21.12% F, Found: 35.22% C, Gd. 4.17% H, N 8.18%, 2.19% S, 20.91% F, 10.12% Example 36 (a) Monobenzyl ester of N trifluoroacetyl · L-glutamic acid

100 g (421.5 mmol) of L-glutamic acid monobenzyl ester are dissolved in a mixture of 1000 ml of trifluoroacetic acid ethyl ester and 500 ml of ethanol and the solution is stirred at room temperature for 24 hours. It is then evaporated to dryness and the residue is recrystallized from diisopropyl ether. Yield 140.47 g (96%) of a colorless crystalline powder.

analysis:

Calculated: C 50.46, H 4.23, F 17.10, N 4.20;

Found: C 51.35, H 4.18, F 17.03, N 4.28.

b) 2-N-Trifluoroacetyl-L-glutamic acid 5-N- (methyl) -N- (2,3,4,6-pentahydroxyhexyl) amide

To a solution of 24.9 g (24.08 mmol) of the title compound of Example 36a, 2? g (24.08 mmol) of N-methylglucamine and 2.77 g (24.08 mmol)

N-hydroxysuccinimide in 150 ml of dimethylformamide is added at 0 ° C with 8.25 g (40 mmol) of N, N-dicyclohexylcarbodiimide. The mixture was stirred at 0 ° C for 3 hours and then at room temperature overnight. The precipitated urea is filtered off and the filtrate is evaporated to dryness in vacuo. The residue is chromatographed on silica gel with dichloromethane-ethanol (20: 1). Yield 109.40 g (89%) of a viscous oil.

analysis:

Calculated: C 51.43, H 5.51, F 13.56, N 6.66.

Found: C 51.22, H 5.41, F 13.40, N. 6.75.

129

c) 2-N-trifluoroacetyl-L-glutamic acid N- (methyl) -N- (2,3,4,5,6-pentahydroxyhexyl) amide

To a solution of 77.33 g (15.15 mmol) of the title compound of Example 36b in 500 mL of ethanol was added 3 g of palladium catalyst (10% Pd / C) and the mixture was hydrogenated at room temperature. The catalyst is then filtered off and the filtrate is evaporated to dryness in vacuo. 43.0 g (100%) of a colorless solid are obtained.

analysis:

H, 5.19; F, 17.26; N, 8.48.

Found: C, 39.84; H, 5.13; F, 17.09; N, 8.68.

d) 2-N-Trifluoroacetyl-L-glutamic acid 5-N- (methyl) -N- (2,3,4,5,6-pentahydroxyhexyl) amide- [1- (4-perfluorooctylsulfonyl) piperazine] amide

To 10.96 g (33.2 mmol) of the title compound of Example 36c a

18.87 g (33.2 mmol) of 1-perfluorooctylsulfonylpiperazine (prepared according to DE 19603033) in 80 mL of THF at 0 C was added ³ 16.42 g (66.4 mmol) EEDQ (ethyl 2-ethoxy-l, 2 (dihydroquinoline-1-carboxylic acid) was added and the mixture was stirred overnight at room temperature. After evaporation in vacuo to dryness, the residue is chromatographed on silica gel with dichloromethane-methanol (20: 1). 28.67 g (92%) of the product are obtained as a colorless solid.

analysis:

Calculated: C 39.61, H 2.89, F 35.66, N 6.19, S 3.54; Found: C, 39.68; H, 2.74; F, 35.81; N, 6.13; S, 3.40.

e) L-glutamic acid 5-N- (methyl) -N- (2,3,4,5, β-pentahydroxyhexyl) amide- [1- (4-perfluorooctylsulfonyl) piperazine] amide

To a solution of 28.36 g (30.22 mmol) of the title compound of Example 36d in 200 mL of ethanol was added ammonia gas at 0 ° C. The mixture was stirred for 4 hours at 0 ° C, evaporated to dryness and residue

130 is mixed with water. The solid was filtered off and dried under vacuum at 50 ° C. Yield 24.19 g (95%) of an amorphous solid.

analysis:

Calculated: C 41.12, H 2.89, F 35.66, N 6.19, S 3.54; Found: C 41.15%, H 2.83%, F 35.78%, N 6.28%, S 3.71%.

f) N- [1,4,7-tris (5-N- (methyl) -N- (2,3,4,5,6-pentahydroxyhexyl) amide- [1- (4-perfluorooctylsulfonyl) piperazine] amide) carboxymethyl) -1,4,7,10-tetraazacyclododecane-10- (pentanoyl-3-aza-4-oxo-5-methyl-5-yl) -L-glutamic acid

20.43 g (24.25 mmol) of the title compound of Example 36e, 2.79 g (24.25 mmol) of N-hydroxysuccinimide, 2.12 g (50 mmol) of lithium chloride and 15.27 g (24.25 mmol) gadolinium complex

1,4,4-Tris (carboxylatomethyl) -10- (3-aza-4-oxo-5-methyl-5-yl) pentanoic acid] -1,4,7,10-tetraazacyclododecane dissolves in 200 g with gentle warming ml of dimethylsulfoxide. At 10 ° C, 8.25 g (40 mmol) of N, N-dicyclohexylcarbodiimide are added and the mixture is stirred overnight at room temperature. The solution was then poured into 3000 ml of acetone and stirred for 10 minutes. The precipitated solid is filtered off and purified by chromatography on RP-18 phase in a water-ethanol-acetonitrile (gradient) system. 23.45 g (75%) of a colorless solid are obtained, containing 11.0% water.

Analysis (calculated on the anhydrous substance):

Calculated: C 34.41, H 3.83, F 23.13, N 9.03, S 2.30,

II, 26% Gd;

Found: C 34.34, H 3.98, F 23.29, N 9.19, S 2.15, Gd 11.07.

Example 37

a) 6-N-Benzyloxycarbonyl-2-N- [1-O-α-D-carbonylmethyl- (2,3,4-tri-O-benzylbenzylglucuronate) -L-lysine- [1- (4-perfluorooctylsulfonyl) piperazine] amide

131

To a solution of the title compound of Example 1c, 100.0 g (120.4 mmol), 73.77 g (120.4 mmol) of 1-O-α-D-carboxymethyl-2,3,4-tri-O- benzyl ester. of benzylglucurone and 13.86 g (120.4 mmol) of N-hydroxysuccinimide in 500 ml of dimethylformamide at 0 ° C are added 41.27 g (200.0 mmol) of N, N-dicyclohexylcarbodiimide. The mixture was stirred at 0 ° C for 3 hours and then at room temperature overnight. The precipitated urea is then filtered off and the filtrate is evaporated to dryness in vacuo. The residue is chromatographed on silica gel with dichloromethane-ethanol (20: 1). 147.58 g (86%) of the product are obtained as a viscous oil.

analysis:

Calculated: C 52.25, H 4.31, N 3.93, F 22.66, S 2.45; Found: C 52.38, H 4.17, N 4.12, F 22.78, S 2.39.

b) 2-N- [1-O-α-D-carbonylmethyl-glucuronic acid] -L-lysine- [1- (4-perfluorooctylsulfonyl) piperazine] amide

To a solution of 142.52 g (100.0 mmol) of the title compound of Example 37a in 2000 mL of ethanol was added 10.0 g of palladium catalyst (10% Pd / C) and the mixture was hydrogenated at room temperature for 12 hours. The catalyst is then filtered off and the filtrate is evaporated to dryness in vacuo. Yield 93.06 g (100%) of a colorless solid.

analysis:

Calculated: C 33.56, H 3.36, N 6.02, S 3.45, F 34.71; Found: C, 33.31; H, 3.42; N, 6.04; S, 3.40; F, 35.51.

c) 6-N- [1,4,7-Tris (carboxylatomethyl)] - 1,4,7,10-tetraazacyclododecane-10-Ν- (pentanoyl-3-aza-4-oxo-5-methyl-5- yl)] - 2-N- [1-O-α-D-carbonylmethyl-glucuronic acid] -L-lysine- [1- (4-perfluorooctylsulfonyl) piperazine] amide, gadolinium complex, sodium salt

50.76 g (54.55 mmol) of the title compound of Example 37b,

6.28 g (54.55 mmol) of N-hydroxysuccinimide, 4.62 g (109.0 mmol)

132 lithium chloride and 34.35 g (54.55 mmol) of 1,4,7-tris (carboxylatomethyl) -10- (carboxy-3-aza-4-oxo-5-methylpent-5-yl) gadolinium complex -1,4,7,10-tetraazacyclododecane was dissolved in 400 ml of dimethylsulfoxide with gentle heating. Add at 10 ° C

16.88 g (81.8 mmol) of N, N-dicyclohexylcarbodiimide and the mixture was stirred overnight at room temperature. The solution was then poured into 3000 ml of acetone and stirred for 10 minutes. The precipitated solid is filtered off and purified by chromatography on RP-18 phase in a water-ethanol-acetonitrile (gradient) system. Yield 37.149 g (88.0%) of a colorless solid containing 8.6% water.

Analysis (calculated on the anhydrous substance):

Calculated: C 34.53, H 3.80, N 8.05, Na 1.47, S 2.05, F 20.63, Gd 10.05;

Found: C, 34.38; H, 3.95; N, 8.19; Na, 1.63; S, 2.15; F, 20.83; Gd, 10.14.

Example 38

a) 6-N-Benzyloxycarbonyl-2- [2- (N-ethyl-N-perfluorooctylsulfonyl) amino] acetyl-L-lysine

To 31.820 g (113.5 mmol) of 6-N-benzyloxycarbonyl-L-lysine and 66.42 g (113.5 mmol) of 2- (N-ethyl-N-perfluorooctylsulfonyl) aminoacetic acid (prepared according to DE 196 03 033) ) in 300 ml of tetrahydrofuran at 0 ° C was added 49.46 g (200.0 mmol) of EEDQ (2-ethoxy-1,2-dihydroquinoline-1-carboxylic acid ethyl ester) and the mixture was stirred overnight at room temperature. It is then evaporated to dryness and the residue is chromatographed on silica gel with dichloromethane-methanol (20: 1). 55.79 g (58%) of the product are obtained as a colorless solid.

analysis:

H, 3.09; N, 4.96; N, 38.11. Found: C, 36.85; H, 3.19; N, 4.87; F, 38.28.

F, 3.78% S; 3,95% S.

133

b) 6-N-Benzyloxycarbonyl-2-N- [2- (N-ethyl-N-perfluorooctylsulfonyl) amino] acetyl-L-lysine-N-methyl-N- (2,3,4,5,6-pentahydroxyhexyl) ) amid

To a solution of the title compound of Example 38a (51.02 g, 60.2 mmol), N-methylglucamine (11.75 g, 60.2 mmol) and 6.93 g (60.2 mmol) of

N-hydroxysuccinimide in 250 ml of dimethylformamide is added at 0 ° C 20.64 g (100.0 mmol) of N, N-dicyclohexylcarbodiimide. The mixture was stirred at 0 ° C for 3 hours and then at room temperature overnight. The precipitated urea is filtered off, the filtrate is evaporated to dryness in vacuo and the residue is chromatographed on silica gel with dichloromethane-ethanol (20: 1). 53.05 g (86%) of the product are obtained in the form of a viscous oil.

analysis:

Calculated: C 38.68, H 4.03, N 5.47, F 31.52, S 3.13; Found: C 38.49, H 4.17, N 5.32, F 31.70, S 3.29.

c) 2-N- [2- (N-ethyl-N-perfluorooctylsulfonyl) amino] acetyl-L-lysine-N-methyl-N- (2,3,4,5,6-pentahydroxyhexyl) amide

To a solution of 102.48 g (100.0 mmol) of the title compound of Example 38b in 2000 mL of ethanol was added 10.0 g of palladium catalyst (10% Pd / C) and the mixture was hydrogenated at room temperature for 12 hours. The catalyst is then filtered off and the filtrate is evaporated to dryness in vacuo. Yield 89.06 g (100%) of the product as a colorless solid.

analysis:

Calculated: C 33.72, H 3.96, N 6.29, S 3.60, F 36.26; Found: C, 33.91; H, 3.82; N, 6.14; S, 3.47; F, 36.31.

d) 6-N- [1,4,7-Tris (carboxylatomethyl)] - 1,4,7,10-tetraazacyclododecane-10-Ν- (pentanoyl-3-aza-4-oxo-5-methyl-5- yl)] - 2-N- [2- (N-ethyl-N-perfluorooctylsulfonyl) amino] acetyl-L-lysine-N-methyl-N- (2,3,4,5,6-pentahydroxyhexyl) amide, gadolinium complex

134

48.58 g (54.55 mmol) of the title compound of Example 38c,

6.28 g (54.55 mmol) of N-hydroxysuccinimide, 4.62 g (109.0 mmol) of lithium chloride and 34.35 g (54.55 mmol) of the gadolinium complex 1,4,7-tris (carboxylatomethyl) - 10- (Carboxy-3-aza-4-oxo-5-methyl-pent-5-yl) -1,4,7,10-tetraazacyclododecane was dissolved in 400 ml of dimethyl sulfoxide with gentle heating. Add at 10 ° C

16.88 g (81.8 mmol) of N, N-dicyclohexylcarbodiimide and the mixture was stirred overnight at room temperature. The solution was then poured into 3000 ml of acetone and stirred for 10 minutes. The precipitated solid is filtered off and purified by chromatography on RP-18 phase in a water-ethanol-acetonitrile (gradient) system. 73.27 g (89.4%) of the product are obtained as a colorless solid which contains 8.6% water.

Analysis (calculated on the anhydrous substance):

Calculated: 35.18% C, 4.23% H, 4.23% N, 2.13% S, 21.50% ¾, 10.47% Gd, ·

Found: C 35.28, H 4.15, N 4.19, S 2.18, F 21.33, Gd 10.61.

Example 39

Contents of the contrast agent of Example 1 of the invention in various organs (including tumor enrichment and lymph nodes) after intravenous administration in rats with prostate cancer

Rats (Cop-Inzucht who were implanted i.m. of Dunning R3327 MAT-Lu prostate carcinoma 12 days ago) received intravenous administration of the resultant compound of Example 1 at a dose of 225 pmol of total gadolinium / kg body weight. The metal content of the various organs, both tumor and lymph nodes (pooled as mesenteric and peripheral lymph nodes) is then determined 10 minutes, 1 hour and 24 hours after application (mean ± standard deviation, n = 3). The results are shown

135 in Table 1.

Table 1

The resulting compound of Example 1 10 P concentration Gd. i. [Μιηοΐ / ΐ] 10 min % of total dose tissue 24 h P · i min . i. 1 h p 24 h p . i. p. i. 1 h p . i. liver 387 ± 26 364 ± 8 74 6 i 34 5.46 ± 0.16 5.81 + 0.16 U, 65 ± 0.97 spleen 54 8 ± 22 487 ± 25 645 ± 27 0,39 ± 0.03 0,39 ± 0.02 0,57 ± 0, 03 pancreas 229 ± 27 199 + 30 130 ± 13 0,26 ± 0, 05 0,21 + 0, 05 C, 17 ± 0.02 kidney 208 1 + 537 883 ± 94 1178 ± 139 5.02 + 1.29 2.15 ± 0.23 2,97 ± 0.21 lungs 837 ± 32 658 ± 29 370 ± 34 1.69 + 0, C 6 1.38 + 0, 08 0.73 ± 0.04 Heart 438 ± 29 289 ± 24 131 + 9 0.46 + I-1 about about 0,31 ± 0, 03 0, 14 ± 0.02 Brain 47 ± 13 26 i 5 14 ± 2 0.15 ± 0.03 0,08 ± 0.02 0,04 + 0, 00 Muscle '* 99 + 5 78 ± 1 36 ± 1 0.11 ± 0.03 0,09 ± 0, 03 0,04 ± 0.00 i.'áčor 185 ± 36 184 i 13 199 ± 19 0,23 ± 0, 10 0,21 + 0, 02 0,31 + 0.01 thigh 184 ± 4 127 + 9 87 ± 6 0,65 ± 0.01 0.46 ± 0, 03 C, 31 ± 0, 03 bone Mes. LU 359 ± 72 697 ± 42 854 ± 135 0, 11 + 0.04 0,24 ± 0, 02 0, 32 ± 0.04 Feathers. LU 229 ± 15 436 + 44 373 ± 24 0.10 ± 0.01 0, 20 ± 0.03 0, 13 ± 0.01 stomach 231 ± 10 219 i 46 138 ± 9 0,57 ± 0.04 0,54 ± 0, 12 C, 37 ± 0.06 (Emptying.) intestine 342 ± 16 409 ± 67 243 ± 22 2.91 + 0.18 3.41 i 1.02 2.14 ± 0, 14 (Emptying.) blood ' 16S 5 ± 110 825 ± 67 214 ± 9 42,95 ± 2.59 21.47 ± 1.78 1, 73 ± 0.03 Rest - _ __ __ _ - 225 ± 31 30,33 ± 4.05 the body Urine 0 - 2 4 h - - - - _ - 94 ± 20 20.21 ± 4, 41 Stolica 0 - - _ __ __ _ - 3128 ± 204 21.33 + i, 46 24 h

* 58 ml blood / kg body weight, ** just an aliquot of right tibial tissue, *** organs total 10 min and 60 min p.i. without other body, **** other body including residual blood.

Example 40

Imaging of lymph nodes (MRT) following intravenous administration of the contrast agent of Example 1 of the invention in rabbits with VX2-tumor

Figure 1 shows MR imaging of iliac lymph nodes in rabbits with i.m. implanted with VX-2 tumor, from pre-administration to 24 hours after intravenous administration

136 of the title compound of Example 1 in an amount of 200 μπιοί Gd / kg body weight. Τχ-weighted gradient echo-frames (1.5 T; sequence: MPRange; TR 11.1 ms, TE 4.3 ms, and 15 ° C) show a strong signal increase in healthy lymph node tissue. Non-signal-enhancing areas in lymph nodes are diagnosed as metastases, which is also confirmed by histological (H / E-staining of lymph node sections). Surprisingly, after a prolonged period (24 hours) of contrast medium administration, the reversed behavior of the signal is observed: The signal in healthy lymph node tissue is reduced, while the metastasis signal now shows a distinct increase.

Surprisingly, a clear increase in the signal in the primary tumor (especially its periphery) can be observed immediately after application. Later (24 hours after application) this increase also spreads to the center of the tumor.

Example 41

Imaging (MRT) of a heart attack after intravenous administration of the contrast agent of Example 1 according to the invention in rats

Figure 2 shows the MR representation of the heart (in vivo and post mortem) 24 hours after intravenous administration of the contrast agent of Example 1 at a dose of 100 pmol Gd / kg body weight in acute induced heart attack rats. T _: -weighted spin-echo-frames (1.5 T; TR: 400 ms, TE: 6 ms; NA: 4; matrix: 128 * 128; layer thickness: 2.5 mm) show strong signal growth in the heart attack area . Successful induction of acute myocardial infarction is confirmed by NBT-staining.

Claims (4)

PATENT CLAIMS 1. Perfluoroalkylated complexes containing sugar moieties of formula I (K)? - G - (ZR ^f ) _m (YR) o (D where R is a monosaccharide or oligosaccharide residue linked through the 1-OH or 1-SH position, R ^f is a perfluorinated, straight or branched hydrocarbon chain of the formula -C _n F ₂ n E _f where E is a terminal fluorine, chlorine, bromine, iodine or hydrogen atom, n equals a number from 4 to 30, K is a metal complex of formula II R ² R ³ O (II) where R ¹ is a hydrogen atom or a metal ion equivalent having atomic number 21-29, 31-33, 37-39, 42-44, 49 or 57-83, provided that at least two R ¹ are metal ion equivalents, R ² and R ³ independently of one another are hydrogen, C 1 -C 4 -alkyl, benzyl, phenyl, -CH ₂ OH or -CH ₂ OCH ₃ , and 138 U is -C ₆ H 2 -O-CH ₂ -? -, - (CH ₂ ) 1-5 -cu, phenylene, -CH ₂ NHCO-CH ₂ -CH (CH ₂ COOH) -C ₅ H ₄ - <o-, -C 5 H ₄ - (OCH ₂ CH ₂ ) 0-1 -N (CH ₂ COOH) -CH ₂ - (or C 1 -C 4 -alkylene or C 7 -C ₁₂ -C) ₆ H ₄ -O-, which latter two groups are optionally interrupted by one or more oxygen atoms, one to three groups -NHCC-, one to three groups -CONH-, and / or substituted by one to three groups - _(CH2) 0-5COOH, where ω is the point of attachment to -CO-, or is a radical of formula III wherein R ¹ is as defined above, R ⁴ is a hydrogen atom or a metal ion equivalent defined for R ¹ and U ¹ is -C 8 H -O-CH ₂ -o, where ω is the point of attachment to -CO- or is the remainder of the general formula of formula IV 139 (IV) wherein R ¹ and R ² are as hereinbefore defined or is a radical of formula VA or VB / COOR ¹ - / - N ROOC / \ ../- COOR ¹ COOR ¹ ABOUT II COOR COOR (UK) COOR ^{1 -} COOR ¹ wherein R ¹ is as defined above, or is a radical of formula VI ROOC - '\ / - \ ./—CO— / N N (j ROOC-COOR ¹ wherein R ¹ is as defined above, or is a residue of formula VII (VI) 140 ROOC ROOC ROOC ROOC (v: i) wherein R ¹ is as defined above and U ¹ is -C 5 H ₄ -O-CH ₂ -o, where ω is the point of attachment to -CO-, or is a radical of formula VIII wherein R ¹ is as defined above, and in the K moiety the free acid groups optionally present may optionally exist as salts of organic and / or inorganic bases or amino acids or amino acid amides, G when K is a metal complex II to VII, is at least three times a functionalized moiety selected from the group of the following moieties a) to j) H coa-N- _{(CH2) 4-C-co-yl} a) ^H NH and β 141 b) H Y ^ CO-CHCH 3 -N-β 1 H NH k <x 142 NH - CO - (CH3) nN2 ⁴ H N ^ Y NH f) N-CO-C- (CH3) nNv4 and HI ²⁴ H NH ί NHI from NH-CO-CH- (CH ₂ ) ₄ -NH- g) N-CO-CH- _(CH2) -NH- ^ Whose NH Y ^is NH-CO-CH (CH ₂₎ 3 NH NH ^ CX ^ N _{(CH2) 4} <N ^{H H} WHAT h) CO-C- (CH ₂ ) ₄ -NI rf NH β 143 1) α ^ ΝΗ — CH — CO— ^{and V} ~ 7 Á β G, when K is a metal complex VIII, is at least three times a functionalized moiety selected from k) or 1), ^k; pv-NH- (CH ₂ ) -CH-CO 2 a NH i Y 1) N, CO-CH-CH-CH-CO ^ Y ² L ² NH i β where a is the point of attachment of residue G to the complex K, β is the point of attachment of residue G to residue Y and γ is the point of attachment of residue G to residue Z, Y is -CH 2 -, δ- (CH 2) n -CO-β (where n = 1-5), δ-ΟΗ ₂ -ΟΗΟΗ-ΟΟ-β, or δ-CH (CHOH-CH ₂₎ OH) -CHOH-CHOH-CO-β, where δ is the point of attachment to the sugar residue R and β is the point of attachment to the residue G, 144 Z is y-n N-SC ₂ -e y-COCH ₂ -N (C ₂ H ₅ ) -5O ₂ -e, y-COCK ₂ -O- (CH ₂ ) ₂ -SO ₂ —ε, O or γ-NKCH ₂ CH ₂ -O-CH ₂ CH ₂ -S, where γ is the point of attachment of Z to the residue G and ε is the point of attachment of Z to the perfluorinated residue R ^f , and M are independently an integer of 1 or 2, and p is an integer of 1 to 4. Metal complexes according to claim 1, characterized in that the equivalent of the metal ion R ¹ is an element with atomic number 21-29, 33, 42, 44 or 57-83. Metal complexes according to claim 1, characterized in that the equivalent of the metal ion R ¹ is an element with atomic number 27, 29, 31-33, 37-39, 43, 49, 62, 64, 70, 75 and 77. Metal complexes according to one of Claims 1 to 3, characterized in that R is a monosaccharide radical having 5 to 6 carbon atoms or a desoxy compound thereof, in particular glucose, mannose or galactose. 145 5. Metal complexes according to one of claims 1 to 4, hearing that K is metal complex general of formula II. 6. Metal kc mplexes according to claim 5 mark your eyes by that R ³ and R ³ independently of one another himself are hydrogen or C 1 -C 6 -alkyl. 7. Metal complexes according to one of claims 1 until 6. in y z n a - hearing that E vo general formula -C _: rJ2nE is fluorine atom. 8. Metal complexes according to one of claims 1 until 7 in y z n a - hearing that G vo general pattern: T means lysine residue (a) or (b). 9. Metal complexes according to one of claims 1 until 8 in y z n a - characterized in that Z in the general formula I is the radical y-n-s ₂ -e wherein γ is the point of attachment of Z to the radical G and ε is the point of attachment of Z to the per fluorinated radical R ^f . Metal complexes according to one of claims 1 to 9, characterized in that Y in the general formula Ϊ is δ-ΟΗζΟΟ-β, where δ is the point of attachment to the sugar moiety R and β is the point of attachment to the moiety G. Metal complexes according to one of Claims 1 to 10, characterized in that U in the metal complex K is -CH ₂ - or -C ₅ H ₄ -CH ₂ -, wherein ω is the point of attachment to -CO- . 146 Use of the metal complexes according to claim 2 for the preparation of contrast media for use in NMR and X-ray diagnostics. Use of metal complexes according to claim 12 for the preparation of contrast agents for imaging infarction and necrosis. Use of metal complexes according to claim 3 for the preparation of contrast agents for application in radiodiagnostics and radiotherapy. Use of metal complexes according to claim 2 for the preparation of contrast agents for lymphography for diagnosing changes in the lymphatic system. Use of metal complexes according to claim 2 for the preparation of contrast agents for application in indirect lymphography. Use of metal complexes according to claim 2 for the preparation of contrast agents for application in i.v. lymphography. Use of metal complexes according to claim 2 for the preparation of contrast means for imaging the vascular bed. Use of metal complexes according to claim 2 for the preparation of contrast agents for tumor imaging. Use of metal complexes according to claim 2 for the preparation of contrast media for imaging the abnormal permeability of capillaries. Pharmaceutical compositions comprising at least one physiologically acceptable compound according to claims 1 to 11, case 147, not with conventional galenic additives. 22. A method for the production of perfluoroalkyl-containing complexes with sugar radicals of general formula I (K) i - G - (ZR ^f) _n and clay) (YR) p wherein A is a metal complex of formula II to VII as claimed in claim 1, G is of formula a) to j) according to claim 1 and Y, Z, R, R ^f , m, p and p have the meanings according to claim 1, characterized in that, in a manner known per se, a carboxylic acid of the formula IIIa wherein R 1 is a metal ion equivalent having atomic numbers 21-29, 31-33, 37-39, 42-44, 49 or 57-83, or a carboxyl protecting group, and R ² , R ³ and U have the previously defined meanings, or carboxylic acid an acid of formula IIIa 148 wherein R ⁴ , R ⁵ and U ¹ are as defined above, or a carboxylic acid of formula IVa wherein R ⁵ and R ² are as defined above, or a carboxylic acid of formula Va or Vb From N-coor ⁵ -N R OOC / .n; -coor ³ ' -COOR ³ HOOC 'x ^ _N ^ COOR Z''- COOR ⁵ A / 5 -COOR -COOR ⁵ Va) where R ⁵ is as defined above, Vb) or a carboxylic acid of the formula VIa 149 R ^S OOC R ^S OOC CO-OH COOR ^S (VIa) wherein R ⁵ is as hereinbefore defined, or a carboxylic acid of formula (VIIa) U 1 OH (VIIa) wherein R ⁵ and U ¹ have the previously given meanings, optionally in activated form, are subjected to a condensation reaction with an amine of formula IX H - G - (ZR ^f) _m and (ix) (YR) p in which G, R, R ^f, Y, Z, maps are as defined above, optionally removing the optionally present protective groups to form a metal complex of formula I or when R ⁵ is a protecting group, after cleavage of these protective groups in a subsequent step in a way known per se, is reacted with at least one metal oxide or metal salt of atomic numbers 21-29, 31-33, 37-39 , 42-44, 49, or 57-83, and finally, if desired, any acidic hydrogen atoms present are replaced by cations of inorganic and / or organic bases, amino acids or amino acid amides. 150 A process for the preparation of perfluoroalkylated complexes containing sugar moieties of the formula I (K) 1 - G - (Z - R ^f ) _m wherein K is a metal complex of the general formula VIII according to claim 1, G is defined by formulas k) or 1) and wherein Y, Z, R, R, ^f , m, p, 1 are as defined in claim 1, characterized in that, in a manner known per se, an amine of formula VIIa wherein R ⁵ is an atomic metal ion equivalent 21-29, 31-33, 37-39, 42-44, 49 or 57-83, or a carboxyl protecting group, is subjected to a condensation reaction with a carboxylic acid, optionally activated, of formula X OH - C - (ZR ^f ) _m I (X) (YR) p in which G, R, R ^f, Y, Z, maps are defined in the claims above, optionally removing the optionally present protective groups to provide a metal complex of general formula I or when R ⁵ is a protecting group, after cleavage of these protective groups in a subsequent step, it is reacted in a manner known per se with at least one metal oxide or metal salt of the element number 21-29, 31-33, 37-39, 42-44, 49 or 57-83, then, if desired, any acidic hydrogen atoms present are replaced with cations of inorganic and / or organic bases, amino acids or amino acid amides. 1/2 FIG. 1 The title compound of Example 1: 200 pmol / kg, i.v .; 3D-T1-MPRage, TR / TE 11.1 / 4.3ms, and 15 °; (arrow: metastasis; T: tumor) before administration 30 min. p.i. 60 min. p.i. 24 hours p.i. The title compound of Example 1: 200 pmol / kg, i.v .; TR / TE 11.1 / 4.3 ms, and 15 °; primary tumor 2/2) BR. 2 24 hours p.i., in vivo, axial 24 hours p.i., post mortem, axial 24 hours p.i., post mortem coronary The title compound of Example 1, 100 pmol / kg i.v .; SE, TR / TE 400/6 ms; arrow: myocardial infarction