Blood depot agents for making nuclear magnetic resonance diagnosis

FIELD: pharmaceutical engineering; medical engineering.

SUBSTANCE: method involves carrying out nuclear magnetic resonance tomography of human or animal blood circulation system containing chelating ion complexes of bivalent and valence three paramagnetic metals of (I)X-L-Y formula, where X is the polyamide carbonyl ligand residue and Y is the gallic acid derivative, .

EFFECT: high accuracy of diagnosis.

28 cl

 

The present invention relates to new complexes of metal ions conjugates of bile acids with molecules possessing chelating activity, as contrast agents for use in diagnostic methods, known as “magnetic resonance imaging”, in particular as agents for blood depot.

The complexes formed by chelating agents and suitable metals, already used as contrast agents in the following ways diagnostics: imaging using x-rays, imaging using nuclear magnetic resonance imaging (M.R.I.) (NMR-tomography and scintigraphy.

In particular, in medical diagnosis using NMR imaging (M.R.I.), widely recognized as a powerful diagnostic method in clinical practice (Stark, D.D., Bradley, W.G., Jr., Eds. "Magnetic Resonance Imaging", The C.V. Mosby Company, St. Louis, Missouri (USA), 1988), used mainly paramagnetic pharmaceutical composition, preferably containing chelate complexes of ions of two or trivalent paramagnetic metal polyaminopolycarboxylic ligands and/or their derivatives or analogues.

Images obtained essentially at the expense of the NMR signal of water protons, are the result of complex interactions between various parameters such as the density of protons and the relaxation time T1and T2. Strengthening the contras is and can be achieved through the introduction of exogenous chemical substances, which significantly alter the resonant characteristics of the neighboring water protons (see Lauffer, R.B. Chem. Rev. 1987, 87, 901).

Paramagnetic contrast agents used for NMR tomography, change the relaxation time of the protons of water present in the tissues in which these contrast agents are concentrated, and therefore they enhance the contrast between different tissues or between healthy and diseased tissue.

Paramagnetic gadolinium complexes were the purpose of the research, publications and patents due to its high ability to reduce the relaxation time of the protons of the neighboring water molecules in the dipolar interactions.

Some of them are represented in clinical practice as M.R.I. contrast agents:

Gd-DTPA, gadolinium complex of N-metilglyukaminovoy salt diethylenetriaminopentaacetic acid, MAGNEVIST®; Gd-DOTA, gadolinium complex of N-metilglyukaminovoy salt of 1,4,7,10-tetraazacyclododecane-1,4,7,10-tetraoxane acid, DOTAREM®; Gd-HPDO3A, gadolinium complex of [10-(2-hydroxypropyl)-1,4,7,10-tetraazacyclododecane-1,4,7-trioxanes acid, PROHANCE®; Gd-DTPA-BMA, gadolinium complex of bis(methylamide) diethylenetriaminepentaacetic acid, OMNISCAN®.

The above contrast agents are commercially available and are intended for wide use. In fact, after the introduction to get M.R.I. specified to strastnye agents diffuse in the blood and in the extracellular areas of the various parts of the body, before they hatch. Therefore, in this respect they are similar iodirovannoi compounds, which are used in medicine for diagnosis using x-rays.

Currently, health workers need of such contrast agents, which are designed for study of specific organs, or for imaging of the vascular system, which cannot be accurately identified using the products commercially available at the present time. The initial approach to obtain the latter consists in the covalent binding contrast agent with macromolecules, such as proteins, or imprisonment inside their stable aggregates of molecules such as liposomes, or using so-called super-paramagnetic particles.

So, for example, gadolinium complex diethylenetriaminepentaacetic acid (Gd-DTPA) was associated with human albumin (HSA), polylysine or dextran (Oksendal A.N. et al., J. Magn. Reson. Imaging, 157, 1993; Rocklage S.M., "Contrast Agents", Magn. Res. Imaging, Mosby Year Book, 372-437, 1992) to reduce or even stop diffusion from the blood into the extracellular fluid, thereby providing a longer stay agent in the circulatory system. This approach, although it was possible to achieve the desired effect, suffers from unacceptable side effects, because the agent is displayed with difficulty.

Another approach is to IP is the use of super-paramagnetic particles, covered with polyethylene glycols or hydrocarbons to reduce their capture in liver endothelial reticulum or in other systems (C. Tilcock, Biochim. Biophys. Acta, 77, 1993; Bogdanoy A.A. et al., Radiology, 701, 1993), thereby increasing the duration of stay of these agents in the blood. In this case also the above mentioned effects are observed, as well as the problems associated with the high cost of production.

Therefore, there is still a need for an effective agent for blood depot, which would have low toxicity and would be a reasonable price.

The present invention relates to a new use as agents for blood depot specially selected compounds that have already been previously disclosed by the applicant in the international patent application WO-A-95/32741, resulting from conjugation of bile acids with a chelating agent, which can form chelate complexes with paramagnetic ions of two or trivalent metals, as well as to new compounds, method of their production and to their use as agents for blood depot.

These compounds demonstrate good hepatobiliary excretion (see P.L. Anelli et al., Acta Radiologica, 38, 125, 1997), which makes them promising contrast agents for imaging of the hepatobiliary system.

Unexpectedly, it was found that specifically the class of these compounds remains in the vascular system quite a long time, that gives the opportunity to use them as contrast agents for imaging of the vascular system, in particular the coronary system.

This effect can clearly be observed in in vivo tests on animals (such as rabbits and monkeys). The consistent finding of these compounds in the vascular system, indeed, it is possible to immediately detect when plotting the values of the relaxation time of protons (1/T1in samples of blood taken with appropriate time intervals after administration of contrast agent.

As complexes of Gd (III) are paramagnetic, high values of 1/T1are proof of the presence in the blood of high concentrations of contrast agent.

The distinction between conventional extracellular contrast agent and blood agent depot is well illustrated in the article Lauffer et al., Radiology, 529, 1998, where are the curves for T1in the blood as a function of time elapsed after administration of contrast agent.

In particular, the complexes of the present invention with the introduction of, for example, to rabbits at a dose comparable with a reasonable safety factor, can cause changes in the velocity of relaxation (defined as Δ 1/T1) in the blood more than 5 sec-110 minutes after the injection, and thus are promising compounds for use in kachestvennosti agents for imaging the circulatory system.

It was found that this type of effect cannot be attributed to the presence of bile acids, but it depends on the chemical structure of the complexes. Apparently, in fact chelating fragment preferably in contact with the steroid skeleton bond in positions 3, 7 or 12, the bile acids.

Indeed, it was proved that any Association between chelating fragment and bile acid including carboxylic group in position 24, leads to the formation of complexes that differ poor consistency in the vascular system.

Therefore, the aim of the present invention is the use as agents for blood depot complexes of compounds of General formula (I) ions with paramagnetic two-trivalent metals selected from the group consisting of Fe(2+), Fe(3+), Cu(2+), Cr(3+), Gd(3+), Eu(3+), Dy(3+), Yb(3+)or MP(2+),

X-L-Y (I)

where

X represents the residue polyaminopolycarboxylic ligand or its derivative selected from the group consisting of:

ethylendiaminetetraacetic acid (EDTA), diethylenetriaminopentaacetic acid (DTPA), 1,4,7,10-tetraazacyclododecane-1,4,7,10-tetraoxane acid (DOTA), 1,4,7,10-tetraazacyclododecane-1,4,7-trioxanes acid (D3), [10-(2-hydroxyprop the l)-1,4,7,10-tetraazacyclododecane-1,4,7-trioxanes acid (HPDO3A), 4-carboxy-5,8,11-Tris(carboxymethyl)-1-phenyl-2-oxa-5,8,11-trustregion-13-OIC acid (WORTH);

Y is derived bile acids selected from the group consisting of residues holeva, chenodeoxycholic, deoxycholic, ursodeoxycholic, lithocholic acid,

as such, as well as funktsionirovanii on the provisions containing a hydroxyl group as a reactive group, regardless of the stereochemistry of the final products, and specified derivative also includes a conjugate acid group in position 24 with taurine and glycine;

L is a chain associated with any position of X, not necessarily including one of the carboxyl groups, which thus turns into an amide group, and C-3, C-7, C-12 Y-positions, and has the following formula (II):

where

m represents an integer from 1 to 10, where values greater than 1 And can have different values,

And represented by the following formula (III):

n and q can be 0 or 1 but cannot both be zero,

R can take values in the interval from 0 to 10

Z represents an oxygen atom or an-NR-group,

where

R represents a hydrogen atom or a (C1-C5) alkyl group, unsubstituted or zameshano the-COOH group.

The most preferred compounds are those in which the spatial chain L have the following General formulas (IIIa) and (IIIb).

Preferable structures in which Z represents an oxygen atom, and L is thus formed due to the hydroxyl groups present in 3, 7, 12 positions, regardless of the stereochemistry of the final products.

Especially preferred compounds of formula (I), in which the residue X is chosen from the group consisting of: EDTA, DTPA, DOTA, D3, WORTH; L is chosen from the group consisting of (IIIa), (IIIb).

Y is preferably selected from the group consisting of residues holeva, deoxycholic, chenodeoxycholic, lithocholic acid, related to L through the amino group in 3-position, and an acid group at position 24 is present as such or in the form of its touringwagon or glycine derivative.

Y can also function in different ways, such as turning one or more hydroxyl groups in ketogroup.

Particularly preferred complexes with paramagnetic ions of the metals listed above are complexes with gadolinium or manganese.

Preferred compounds of General formula (IV), in which in the General formula (I) residue X is DTPA, substituted on the Central chain, and where R1represents a hydrogen atom or-COOH group,

where Y is chosen from the group consisting of residues holeva, deoxycholic, chenodeoxycholic, lithocholic acid, a L is represented by formula (III).

Especially preferred compounds of General formula (IVa):

where R1represents a-COOH group, and Y takes above for compounds of General formula (IV) values, and L is represented by formula (IIIa) or (IIIb).

The following objectives of the present invention are new compounds belonging to the class of compounds of General formula (IVa), as well as methods for their production;

[3β (S)

5β ]-3-[[4-[bis[2-[bis(carboxymethyl)amino]ethyl]amino]-4-carboxy-1-oxobutyl](carboxymethyl)amino]Holan-24-OIC acid

[3β (S)

5β ]-3-[[4-[bis[2-[bis(carboxymethyl)amino]ethyl]amino]-4-carboxy-1-oxobutyl]amino]Holan-24-OIC acid

[3β (S),5β ,

7β ]-3-[[4-[bis[2-[bis(carboxymethyl)amino]ethyl]amino]-4-carboxy-1-oxobutyl]amino]-7-hydroxypoly-24-OIC acid

[3α (S)

5β ]-3-[2-[[5-[bis[2-[bis(carboxymethyl)amino]ethyl]amino]-5-carboxypentyl]amino]-2-oksidoksi]Holan-24-OIC acid

[3β (S)

5β ]-3-[[4-[bis[2-[bis(carboxymethyl)amino]ethyl]amino]-4-carboxy-1-oxobutyl]the Mino]-12-Oklahoman-24-OIC acid

[3β (S), 5β ,

7α ]-3-[[4-[bis[2-[bis(carboxymethyl)amino]ethyl]amino]-4-carboxy-1-oxobutyl]amino]-7-hydroxypoly-24-OIC acid

N2bis[2-[bis(carboxymethyl)amino]ethyl]-N-[(3β , 5β , 7α , 12α )-7,12-dihydroxy-24-oxo-24-[(2-sulfoethyl)amino]Holan-3-yl]-L-glutamine

N2bis[2-[bis(carboxymethyl)amino]ethyl]-N-[(3β , 5β )-24-oxo-24-[(2-sulfoethyl)amino]Holan-3-yl]-L-glutamine

[3β (S), 5β , 12α ]-3-[[4-[bis[2-[bis(carboxymethyl)amino]ethyl]amino]-4-carboxy-1-oxobutyl]amino]-12-hydroxypoly-24-OIC acid

[3β (R), 5β , 12α ]-3-[[4-[bis[2-[bis(carboxymethyl)amino]ethyl]amino]-4-carboxy-1-oxobutyl]amino]-12-hydroxypoly-24-OIC acid

[3β (RS), 5β , 12α ]-3-[[4-[bis[2-[bis(carboxymethyl)amino]ethyl]amino]-4-carboxy-1-oxobutyl]amino]-12-hydroxypoly-24-OIC acid

[3α (S), 5β , 12α ]-3-[[4-[bis[2-[bis(carboxymethyl)amino]ethyl]amino]-4-carboxy-1-oxobutyl]amino]-12-hydroxypoly-24-OIC acid

[3β (RS), 5β , 7α , 12α ]-3-[[4-[bis[2-[bis(carboxymethyl)amino]ethyl]amino]-4-carboxy-1-oxo is util]amino]-7,12-dihydroxyfuran-24-OIC acid

[3α (S), 5β , 7α , 12α ]-3-[[[[5-[bis[2-[bis(carboxymethyl)amino]ethyl]amino]-5-carboxypentyl]amino]carbonyl]oxy]-7,12-dihydroxyfuran-24-OIC acid

[3α (S), 5β ]-3-[2-[[5-[bis[2-[bis(carboxymethyl)amino]ethyl]amino]-5-carboxypentyl]amino]-2-oksidoksi]Holan-24-OIC acid

Other compounds belonging to this class, complexes with gadolinium disclosed in the patent application WO-A-95/32741 presented below:

[3β and (3), 5β , 7α , 12α ]-3-[[4-[bis[2-[bis(carboxymethyl)amino]ethyl]amino]-4-carboxy-1-oxobutyl]amino]-7,12-dihydroxyfuran-24-OIC acid

[3β (S), 5β , 7α , 12α ]-3-[[4-[[5-[bis[2-[bis(carboxymethyl)amino] ethyl]amino]-5-carboxypentyl]amino]-1,4-DIOXOLANYL]amino]-7,12-dihydroxyfuran-24-OIC acid;

Preferred are also compounds of General formula (IVb), which are also DTPA derivatives, substituted in the Central position,

where Y takes above for compounds of formula (IV) values, a L takes values corresponding to formula (IIIa).

Further, the present invention relates to the following new compounds that belong to a class of compounds which deposits of General formula (IVb), and retrieval method:

(3β , 5β , 7α , 12α )-3-[[[bis[2-[bis(carboxymethyl)amino]ethyl]amino]acetyl]amino]-7,12-dihydroxyfuran-24-OIC acid

(3β , 5β -3-[[[[[bis[2-[bis(carboxymethyl)amino]ethyl]amino]acetyl]amino]acetyl]amino]Holan-24-OIC acid

Other compounds belonging to this class, complexes with gadolinium were disclosed in the patent application WO-A-95/32741 presented below:

(3β , 5β , 7α , 12α )-3-[[[[[bis[2-[bis(carboxymethyl)amino]ethyl]amino]acetyl]amino]acetyl]amino-7,12-dihydroxyfuran-24-OIC acid

(3β , 5β , 7α , 12α )-3-[[6-[[[bis[2-[bis(carboxymethyl)amino]ethyl]amino]acetyl]amino]-1-oxohexyl]amino]-7,12-dihydroxyfuran-24-OIC acid

Especially preferred are also compounds of General formula (V), in which in the General formula (I) residue X is DTPA, Y takes on the values specified above for the compounds of General formula (IV), and L is represented by formula (IIIa).

Other compounds belonging to this class, complexes with gadolinium were disclosed in the patent application WO-A-95/32741 presented below:

(3β , 5β , 7α , 12α )-3-[[N-[N-[2-[[2-[bis(carboxymethyl)aminoethyl](carboxymethyl)amino]ethyl]-N-(carboxymethyl)glycyl]glycyl]amino]-7,12-dihydroxyfuran-24-OIC acid

18-[[(3β , 5β , 7α , 12α )-23-carboxy-7,12-hydroxy-24-norolean-3-yl]amino]-3,6,9-Tris(carboxymethyl)-11,18-dioxo-3,6,9,12-tetraisostearate acid

Preferred are also the compounds of formula (VI), in which in the formula (I) residue X presents D3, Y takes on the values specified above for the compounds of General formula (IV), and L is chosen from compounds of formulas (IIIa) and (IIIb).

Among the compounds of formula (VI) is particularly preferred 10-[3-[[(3α , 5β , 7α , 12β )-23-carboxy-7,12-dihydroxy-24-norolean-3-yl]oxy]-2-hydroxypropyl]-1,4,7,10-tetraazacyclododecane-1,4,7-trioxane acid, complex with gadolinium was disclosed in patent application WO-A-95/32741.

Likewise preferred compounds of General formula (VII), in which in the formula (I) residue X presents EDTA, Y takes on the values specified above for compounds of formula (IV), a L corresponds to formula (III).

Particularly preferred complexes of the compounds of the formula (VII) with manganese

Among the compounds of formula (VII) are particularly preferred are the following:

[3α (S), 5β , 12α ]-3-[[[[5-[[2-[bis(carboxymethyl)amino]ethyl](carboxymethyl)amino]-5-carboxypentyl]amino]carbonyl]oxy]-1-hydroxypoly-24-OIC acid

[3β (S), 5β , 7α , 12α ]-3-[[4-[[5-[[2-[bis(carboxymethyl)amino]ethyl](carboxymethyl)amino]-5-carboxypentyl]amino]-1,4-DIOXOLANYL]amino]-7,12-dihydroxyfuran-24-OIC acid

[3α (S), 5β ]-3-[2-[[5-[[2-[bis(carboxymethyl)amino]ethyl](carboxymethyl)amino]-5-carboxypentyl]amino]-2-oksidoksi]Holan-24-OIC acid

[3β (S), 5β , 12α ]-3-[[4-[[2-[[bis(carboxymethyl)amino]ethyl](carboxymethyl)amino]-4-carboxy-1-oxobutyl]amino]-12-hydroxypoly-24-OIC acid

[3β (S), 5β ]-3-[[4-[[2-[[bis(carboxymethyl)amino]ethyl](carboxymethyl)amino]-4-carboxy-1-oxobutyl]amino]-12-Oklahoman-24-OIC acid

Compounds of General formula (I) can be obtained by way of a convergent synthesis, which includes:

1) synthesis funkzionirovanija ligand, i.e. a ligand that can coordinate one ion of a paramagnetic metal, while remaining stably associated with bile acid through a suitable functional group;

2) synthesis funktsionirovanii bile acids;

3) the reaction mix two different synthons;

4) remove any protective group;

5) the formation of the complex ion of a paramagnetic metal; illustrated in detail the above-mentioned patent application WO-A-95/32741.

Some of the preferred methods of obtaining the ligands of the present invention include the formation of amide linkages between the two synthons, and one of them is a precursor chelat forming system of paramagnetic ion (synthon), and the other is the predecessor of the balance of bile acids in the target complex (synthon).

Following methods should not be construed as limiting the methods of obtaining the compounds of the present invention.

Amide bond can be created:

a) interacting synthons And containing carboxyl functionality, synthon In containing a primary or secondary aminophenol;

b) interacting synthons And containing a primary or secondary aminophenol, synthon In containing carboxyl function;

c) through the interaction of DTPA dianhydride (commercially available product) synthon In containing a primary or secondary aminophenol.

A list of some of the little a and b used in the method of the present invention, shown in the following table.

Naturally, the synthons used should be well protected for the groups, which could be a source of unwanted reactions in conditions that are used to create amide of light is I. After the formation of the amide bond between the two synthons may provide one or more stages removal of the protective groups for conservation groups source.

Alternatively, the methods of this type, chelating subunit can be entered in the result of multi-stage reactions, derived from bile acids, as in the case of the synthesis of compounds disclosed in example 3 experimental section, as illustrated in figure 1.

Scheme 1

The present invention relates also to a new method, which is illustrated in the following scheme 2:

Scheme 2

where

R4is aminosidine group;

R5represents a branched or unbranched C1-C10alkyl or aryl,

R2and R3represent independently a hydrogen atom, a branched or unbranched C1-C20alkyl, unsubstituted or substituted aryl groups, or these groups form With3-C10cycle;

moreover, this method uses the reaction of transaminirovania that allows you to save the stereochemistry at the chiral center adjacent to the nitrogen atom source pyrrolidinone, and get a secondary amide. The combined selection of the groups R4and R5important in the sense that splitting should PR is to proceed under different conditions. Possible examples R4are carboxymethylate (Cbz) group, and examples R5are methyl or tert-bucilina group.

This method is usually used for γ -amides glutaminol acid, and it is very useful for producing compounds of the present invention, in particular γ -amides glutaminol acid with 3-amino residues Y, as defined above. In fact, it allows to obtain the target compound, avoiding the use of expensive condensing agents for education γ -amido communication between glutaminol acid and the appropriate amine.

An example of the application of this new method is the synthesis of [3β (S), 5β , 12α ]-3-[[4-[bis[2-[bis(carboxymethyl)amino]ethyl]amino]-4-carboxy-1-oxobutyl]amino]-12-hydroxypoly-24-OIC acid, a common way of receiving which is presented in example 4 experimental section, while the alternative method shown in example 5, and the full scheme of the synthesis is presented below figure 3.

Scheme 3

Similarly receive derived holeva acid already described in patent application WO-95/32741, [3β (S), 5β , 7α , 12α ]-3-[[4-[bis[2-[bis(carboxymethyl)amino]ethyl]amino]-4-carbox the and-1-oxobutyl]amino]-7,12-dihydroxyfuran-24-OIC acid.

Metal ions capable of forming a complex with the chelating agents of General formula (I) are divalent or trivalent ions of elements selected from the group consisting of Fe(2+), Fe(3+), Cu(2+), Cr(3+), Gd(3+), Eu(3+), Dy(3+), Yb(3+)or MP(2+).

As for the diagnostic use of new chelate complexes of the present invention, they can be used as contrast agents, particularly for use as agents for blood depot for diagnosis using NMR imaging.

Complexes usually get in accordance with the method in which the oxide or a suitable salt of the paramagnetic metal dissolved in water or suspended in a water-alcohol solution is added to aqueous or aqueous-alcoholic solution of a chelating agent with stirring and, if necessary, heated moderately, or up to the boiling point, until the completion of the reaction. If the complex is insoluble in the reaction solvent, it is possible to filter. If it is soluble, it can be selected, viparita solvent to obtain a residue, for example, by spray drying.

In that case, if the received complex still contains free acid groups, it is transformed into a neutral salt by reaction with an organic or inorganic base is m, which form physiologically compatible cations in solutions.

To obtain such neutral salts sufficient grounds can be added to the complexes containing free acid groups, in aqueous solution or suspension to neutrality. The resulting solution can then evaporate, or suitable solvent can be added for crystallization of the salt complex.

Preferred inorganic cations, suitable for salt chelate complexes of the present invention include, in particular, ions of alkali or alkaline earth metals such as potassium, sodium, calcium, magnesium, and mixtures thereof. Particularly preferred sodium ion.

Preferred cations derived from organic bases suitable for the above purposes include (among others) the cations of primary, secondary and tertiary amines, such as ethanolamine, diethanolamine, morpholine, glucamine, N-methylglucamine, N,N-dimethylglycine, with the most preferred N-methylglucamine.

Preferred cations are derived from amino acids include, for example, cations of taurine, glycine, lysine, arginine or ornithine.

The alternative to this method is to obtain compositions for injection without isolating the salt complex. In this case, the final solution does not necessarily have to contain free the metal ions, which is toxic to the organism.

This can be checked by titration, for example, with color indicators, such as Xylenol orange. You can also include a stage of purification of the salt complex.

In this type of way chelating agent, a salt or a metal oxide, and any soleobrazutaya Foundation subjected to interaction in stoichiometric ratios in water for injection, and then, after completion of the reaction, pyrogens filtered and the product is distributed in suitable containers, and then carry out thermal sterilization.

Pharmaceutical compositions for injection are usually obtained by dissolving the active ingredient, obtained as above, and excipients in the water of the required purity from a pharmacological point of view, in order to obtain a pharmaceutical composition suitable for enteral or parenteral administration, in molar concentrations in the range of from 0.01 to 1.0. Received a contrast agent properly sterilized.

Contrast agents injected, depending on the diagnostic requirements in doses from 0.01 to 0.3 mmol/kg of body weight.

In principle doses for parenteral administration are in the range from about 0.001 to about 1.0 mmol/kg of body weight. Preferred doses for parenteral administration are in the range from 0.01 to 0.5 mmol/kg of body weight.

On the PS for enteral introduction are usually in the range of from 0.5 to about 10 mmol/kg body weight, preferably from about 1.0 to about 10 mmol/kg of body weight.

New compositions of the present invention demonstrate good tolerability; moreover, their solubility in water is an additional important feature, which makes them particularly suitable for use in nuclear magnetic resonance.

The diagnostic compositions of the present invention is used in the usual way. Songs you can enter the patient, typically a warm-blooded animal, as systematically and surface in the organ or tissue to be visualized using nuclear magnetic resonance.

Circuit analysis and equipment can be found in works such as Stark, D.D., Bradley, W.G., Magnetic Resonance Imaging, Mosby Year Book, St. Louis, Mo, 1992.

Used conditions of experiments will be illustrated in detail in the experimental section.

Experimental section

EXAMPLE 1

The gadolinium complex of 1-deoxy-1-(methylamino)-D-glaciology salt [3β (S), 5β ]-3-[[4-[bis[2-[bis(carboxymethyl)amino]ethyl]amino]-4-carboxy-1-oxobutyl]amino]Holan-24-OIC acid (1:3)

A) Methyl ester [3β (S), 5β ]-3-[[5-(1,1-dimethylmethoxy)-4-[bis[2-[bis[2-(1,1-dimethylmethoxy)-2-oxoethyl]amino]ethyl]amino]-1,5-dioxaphetyl]amino]Holan-24-OIC acid

3.6 g of methyl ether (3β , 5β )-3-Linkoln-24-OIC sour is s (obtained similarly to the way opened in WO-A-95/32741: example 5) (9,24 mmol), 8.5 g of tert-butyl methyl ether N,N-bis[2-[bis[2-(1,1-dimethylmethoxy)-2-oxoethyl]amino]ethyl]-L-glutamato acid (obtained as described in WO-A-95/32741: example 15) (is 11.39 mmol) and 1.64 g diethylthiophosphate (9,39 mmol) is dissolved in 160 ml of DMF. The solution is cooled to 0° C, are added dropwise to 1.28 ml Et3N (9,24 mmol) and the reaction mixture is left for 30 minutes at room temperature. After 1 hour the solution is evaporated under reduced pressure, the residue is mixed with EtOAc, washed with 5% Panso3and then brine. The organic phase is allocated, dried over Na2SO4and then evaporated under reduced pressure. The crude product is purified using flash chromatography, getting 9.5 g of the desired product (8.50 mmol).

Yield: 92%

K.F.: 3,47%

Elemental analysisCHN
% calculated:66,639,745,01
% found:a67,4210,085,07

andafter drying at 120° With vacuum

TLC: Stationary phase: a plate of silica gel 60F 254 Merck; Eluent = 4:6 EtOAc/n-hexane

Detection: 0.5% KMPO4in 1 M NaOH Rf=0,46

Spectra1H-NMR,13C-NMR, IR and MS correspond to the specified structure.

<> C) Methyl ester [3β (S), 5β ]-3-[[4-carboxy-4-[bis[2-[bis(carboxymethyl)amino]ethyl]amino]-1-oxobutyl]amino]Holan-24-OIC acid

To a stirred solution of 9.3 g of compound obtained in stage A) (8,32 mmol)in 50 ml of CH2CL2add to 5.1 ml of CF3COOH (66,6 mmol); after 10 min at a temperature of 0-5° the solution is evaporated. The residue is placed in 50 ml of CF3COOH and after 24 hours at room temperature, add 30 ml of CF3COOH to complete the reaction. After 5 hours the reaction mixture is evaporated and the residue is treated CH2Cl2, viparita every time the solvent under reduced pressure to obtain a powder. The solid product is washed with H2O, filtered and dried, obtaining the desired product (6.9 g; 8,24 mmol).

A yield of 99%. So PL: 205°

K.F.: 7,78%

Elemental analysisCHN
% calculated:60,278,186,69
% found:a59,288,11of 6.68

andafter drying at 120° With vacuum

TLC: Stationary phase: a plate of silica gel 60F 254 Merck

Eluent=6:4:1 l3/Meon/25% NH4OH.

Detection: 0.5% KMPO4in 1 M NaOH Rf=0,28

Spectra1H-NMR,13The NMR, IR and MS correspond to the specified structure.

C) [3β (S), 5β ]-3-[[4-[bis[2-[bis(carboxymethyl)amino]ethyl]amino]-4-carboxy-1-oxobutyl]amino]Holan-24-OIC acid

To a suspension 6,14 g of compound obtained in stage C) (7,33 mmol)in 50 ml of H2O add 50 ml of 1 M NaOH (50 mmol), maintaining the pH value of 13 by using a pH-stat apparatus. After 2 hours at room temperature, the reaction mixture is acidified (pH 0.5) the addition of 12 M Hcl, receiving the suspension, which was filtered, washed with H2O and dried to obtain the desired product (5,64 g; 6,85 mmol).

Yield: 93%. So pl.: 205°

K.F.: 9,02%

Elemental analysisCHNCl, Na
% calculated:59,848,08for 6.81 
% found:a59,568,15to 6.80<0,1

andafter drying at 120° With vacuum

TLC: Stationary phase: a plate of silica gel 60F 254 Merck

Eluent=6:4:1 l3/Meon/25% NH4OH

Detection: 0.5% KMPO4in 1 M NaOH Rf=0,25

Spectra1H-NMR,13C-NMR, IR and MS correspond to the specified structure.

D) gadolinium Complex of 1-deoxy-1-(methylamino)-D-glaciology salt [3β (S), 5β ]-3-[[4-[bis[2-[bicarboxylic)amino]ethyl]amino]-4-carboxy-1-oxobutyl]amino]Holan-24-OIC acid (1:3)

a 4.53 g of compound obtained in stage C), (5.5 mmol), suspended in 50 ml of N2O and solubilizers 10 ml of 2 M aqueous solution of meglumine (20 mmol), obtaining the solution at pH 6.8. After that, within 1 hour, add 11 ml of 0.5 M aqueous solution Gdl3(5.5 mmol), maintaining the pH value of 6.8 by the addition of 6.5 ml of 2 M aqueous solution of meglumine (13 mmol). The reaction course is monitored by capillary electrophoresis. After 2 hours the solution is filtered through a membrane Millipore®filter through nanofilter and evaporated. The residue is dried, obtaining the desired compound (6,15 g; to 4.17 mmol).

Yield: 76%. T square: 220°

K.F.: 8,44%

CE (capillary electrophoresis) analysis: 100% (area %)

Spectra IR and MS correspond to the specified structure.

The following compounds and the corresponding complexes of gadolinium get a similar way:

The gadolinium complex of 1-deoxy-1-(methylamino)-D-glaciology salt [3β (S), 5β , 7β ]-3-[[4-[bis[2-[bis(carboxymethyl)amino]ethyl]amino]-4-carboxy-1-oxobutyl]amino]-7-hydroxypoly-24-OIC acid (1:3);

The gadolinium complex of 1-deoxy-1-(methylamino)-D-glaciology salt [3α(S), 5β ]-3-[2-[5-[bis[2-[bis(carboxymethyl)amino]ethyl]amino]-5-carboxypentyl]amino]-2-oksidoksi]Holan-24-OIC acid (1:3);

EXAMPLE 2

The gadolinium complex of 1-deoxy-1-(methylamino)-D-glaciology salt is [3β (S), 5β ]-3-[[4-[bis[2-[bis(carboxymethyl)amino]ethyl]amino]-4-carboxy-1-oxobutyl]amino]-12-Oklahoman-24-OIC acid (1:3)

A) Methyl ether (3β , 5β )-3-azido-12-Oklahoman-24-OIC acid

12.5 ml of Jones reagent [33.3 mmol Cr (VI)] is added dropwise to a solution of 17.8 g of methyl ether complex (3β , 5β , 12α )-3-azido-12-hydroxypoly-24-OIC acid (41,1 mmol) (obtained analogously to the method described for the methyl ester (3β , 5β , 7α , 12α )-3-azido-7,12-dihydroxyfuran-24-OIC acid, WO-A-95/32741: example 5) in acetone (600 ml) for 90 minutes at room temperature. After 20 hours the mixture is filtered and the solution evaporated. The residue is dissolved in l3(400 ml) and the solution washed with saturated aqueous Panso3then H2O. the Solution is dried and evaporated, to give crude product which is crystallized from 96% EtOH to obtain 14.1 g of the desired product (from 32.9 mmol).

Yield: 84%. TPL: 153°

K.F.:<0,1%

[α ]

20
D
=+83,25 (2,1, l3)

Elemental analysisCHN
% calculated:69,909,159,78
% found:69,98to 9.32RS 9.69

TLC: Stationary phase: a plate of silica gel 60F 254 Merck

Eluent: 8:2 n-hexane /EtOAc

Detection: 0.5% KMPO4in 1 M NaOH Rf=0,43

Spectra1H-NMR,13C-NMR, IR and MS correspond to the specified structure.

C) Methyl ether (3β , 5β )-3-amino-12-Oklahoman-24-OIC acid

A solution of 16.4 g of compound A) (38,2 mmol) in THF (130 ml) hydronaut in the presence of 5% Pd/C (1.6 g) at room temperature and a pressure of 40 bar for 15 hours in an autoclave Parr®. The reaction mixture is filtered (paper and membrane FH 0.5 µm Millipore®) and evaporated. The crude residue purified using flash chromatography, getting to 11.8 g of the desired product (29.2 mmol).

Yield: 77%. TPL:129-130°

K.F.: 1,04%

[α ]

20
D
=+87,8 (with 2.02, l3)

Elemental analysisCHN
% calculated:74,4010,243,47
% found:72,7210,003,35

TLC: Stationary phase: a plate of silica gel 60F 254 Merck

Eluent: 95:5 MeOH/Et3N

Detection: 0,5% KMnO4in 1 M NaOH Rf0,31

Spectra1H-NMR,13C-NMR, IR and MS correspond to the specified structure.

C) Methyl ester [3β (S), 5β ]-3-[[4-[bis[2-[bis[2-(1,1-dimethylmethoxy)-2-oxoethyl]amino]ethyl]amino]-5-(1,1-dimethylmethoxy)-1,5-dioxaphetyl]amino]-12-Oklahoman-24-OIC acid

A solution of DCC (6,24 g, 30.3 mmol) in CH2Cl2(25 ml) is added dropwise during 30 minutes to a solution of 1-(1,1-dimethylethanol) ether N,N-bis[2-[bis[2-(1,1-dimethylmethoxy)-2-oxoethyl]amino]ethyl]-L-glutamato acid (obtained as described in WO-A-95/32741: example 15) (21,5 g; of 28.9 mmol), connection) (11.1 g, 27.5 mmol) and NOT (1-hydroxybenzotriazole) (3,72 g; 27.5 mmol) in CH2Cl2(300 ml) at 0° C in nitrogen atmosphere. The mixture is left to warm to room temperature. After 21 hours the reaction mixture is filtered and the solution was washed with saturated aqueous Panso3then H2Oh and then evaporated. The crude product is purified using flash chromatography, receiving 24.5 g of the desired product (21,7 mmol).

Yield: 79%

[α ]

20
D
=+12,17 (c 2,07, l3)

TLC: Stationary phase: a plate of silica gel 60F 254 Merck

Eluent: 1:1 EtOAc/n-hexane

Detection: 0.5% KMPO4in 1 M NaOH Rf=0,45

Spectra1H-NMR,13C-NMR, IR and MS correspond to the specified structure.

D) [3β (S), 5β ]-3-[[4-[bis[2-[bis(carboxymethyl)amino]ethyl]amino]-4-carboxy-1-oxobutyl]amino]-12-Oklahoman-24-OIC acid

80 ml TFU (triperoxonane acid) (1.0 mol) is added dropwise to a solution of 23.8 g of compound obtained in stage C), (21,0 mmol)in CH2Cl2(50 ml) at 0° C for 1 hour. The reaction mixture was stirred at room temperature, followed by 2 hours and evaporated. The residue is placed in TFU (100 ml, 1.3 mol) and the solution stirred for further 24 hours. Then the reaction mixture is evaporated, add CH2Cl2and again evaporated. The crude residue is dissolved in 150 ml of 1 M NaOH, cooling in a bath with ice, and the solution is then stirred for 15 h (pH 10) at room temperature. Bring the pH of the reaction mixture to 13 by adding 3,30 ml of 30% NaOH and after 4 hours, filtered through a Millipore membrane® HAS 0.45 µm). The filtrate is acidified with 12.5 ml of 30% Hcl and 19 ml of 1 M Hcl to a pH of at least 1.60. The precipitate is filtered off, washed with N2O and dried, getting to 15.8 g of the desired product (to 18.9 mmol).

Output 90%. TPL 172-175°

K.F.: 1,98%

[α ]

20
D
=+43,54 (with 2.02, 1 M NaOH)

HPLC: 97% (area %)

Stationary phase:Bond ECLIPSE XDB-C8 3.5 µm; 150× 4.6 mm
Temperature:40°
Mobile phase:gradient elution
 A=0,017 M H3RHO4, 0.3 mm EDTA in N2About
 In=CH3SP  
Gradient:min%%
 08515
 406535
 506535
Flow rate:1.5 ml/min  
Detection (UV):210 nm
Elemental analysisNNCLNa
% calculated58,847,716,69  
% found56,577,686,370,250,18

TLC: Stationary phase: a plate of silica gel 60F 254 Merck

Eluent: 5:4:2 l3/Meon/25% NH4OH

Detection: 0.5% KMPO4in 1 M NaOH Rf=0,28

Spectra1H-NMR,13C-NMR, IR and MS correspond to the specified structure.

E) gadolinium Complex of 1-deoxy-1-(methylamino)-D-glaciology salt [3β (S), 5β ]-3-[[4-[bis[2-[bis(carboxymethyl)amino]ethyl]amino]-4-carboxy-1-oxobutyl]amino]-12-Oklahoman-24-OIC acid (1:3)

49,0 ml 0,918 M aqueous solution of meglumine (45,0 mmol) is added dropwise to a suspension of 14.0 g of compound obtained in stage D) at 16.7 mmol), in N2O (100 ml) at room temperature, obtaining a clear solution (pH 6.5). To it is added dropwise to 31.6 ml 0,503 M aqueous solution of GdCl3(15.9 mmol), maintaining a pH of 6.5 by the addition of 55.7 ml 0,918 M aqueous solution of meglumine (51,1 mmol) using a pH-stat. At the end of the addition the mixture is filtered through a membrane Millipore® (HAWP 0.45 µm), passed through nanofilter, pH adjusted to 7.0 by the addition of 0.100 ml 0,918 M aqueous solution of meglumine (0,092 mmol) and evaporated. The residue is dried, getting to 22.0 g of the desired product (14.0 mmol).

Yield 84%. TPL 100-105°

K.F.: 5,06%

HPLC analysis: 97% (area %)

Stationary phase: HYPURITY™ Elite C18 5 μm; 250× 4.6 mm column Hypersil;

Spectra IR and MS correspond to the specified structure.

EXAMPLE 3

The gadolinium complex of 1-deoxy-1-(methylamino)-D-glaciology salt [3β , 5β , 7α , 12α ]-3-[[[bis[2-[bis(carboxymethyl)amino]ethyl]amino]acetyl]amino]-7,12-dihydroxyfuran-24-OIC acid (1:2)

A) Methyl ester [3β , 5β , 7α , 12α ]-3-[[[bis[2-[bis[2-(1,1-dimethylmethoxy)-2-oxoethyl]amino]ethyl]amino]acetyl]amino]-7,12-dihydroxyfuran-24-OIC acid

24.8 g of methyl ether (3β , 5β , 7α , 12α )-3-[(aminoacetyl)amino]-1,12-dihydroxyfuran-24-OIC acid (obtained according to the method, opened in WO-A-95/32741: example 5) (51,9 mmol) are suspended in a mixed solution of 38.7 g of 1,1-dimethylethylene ether N-(2-bromacil)-N-[2-(1,1-dimethylmethoxy)-2-oxoethyl]glycine (obtained by the method, opened in WO-A-95/32741: example 15) (110 mmol) in 390 ml of CH3The JV. Adding 245 ml of 2M phosphate buffer pH 8 results in a two-phase solution, which is intensively stirred at room temperature for 144 hours. The organic phase is isolated and evaporated, the remaining oil is dissolved in 250 ml of CH2Cl2. The solution is washed with N2O, dried (Na2SO4) and evaporated. The crude product is purified using flash chromatography (eluent=95:5 CHCl3/CH3IT), getting the right product (24.8 g; 24,3 m is ol).

Exit; 47%

[α ]

20
D
=+9,45 (1.5, l3)

Elemental analysisCHN
% calculated:64,68for 9.475,49
% found:64,559,445,46

TLC: Stationary phase: a plate of silica gel 60F 254 Merck

Eluent: 88:12 l3/Meon

Detection: 0.5% KMPO4in 1 M NaOH Rf=0,57

Spectra1H-NMR,13C-NMR, IR and MS correspond to the specified structure.

B) (3β , 5β , 7α , 12α )-3-[[[bis[2-[bis(carboxymethyl)amino]ethyl]amino]acetyl]amino]-7,12-dihydroxyfuran-24-OIC acid

318 ml of 2 M aqueous solution of LiOH (636 mmol) is added dropwise over 15 minutes to a solution of compound obtained in stage A) (21,6 g, 21.1 mmol)in 310 ml of EtOH. 23 hours EtOH is evaporated and the reaction mixture is stirred for further 2 hours. The solution is added dropwise in 255 ml of 2.6 M Hcl and bring the pH to 1.4 30% NaOH. After 1.5 hours the precipitate is filtered off, washed with 300 ml of 0.1 M Hcl and dried, obtaining the desired product (13.1 g; of 16.7 mmol).

C) gadolinium Complex of 1-deoxy-1-(methylamino)-D-glaciology salt (3β 5β , 7β , 12α )-3-[[[bis[2-[bis(carboxymethyl)amino]ethyl]amino]acetyl]amino]-7,12-dihydroxyfuran-24-OIC acid (1:2)

11.3 g of the compound obtained in stage C) (13,8 mmol), suspended in 40 ml of H2O and dissolved by the addition of 44.7 ml of 1 M aqueous solution of meglumine (44,7 mmol) up to pH 6. To the mixture is added dropwise to 13.7 ml of 1 M aqueous solution of GdCl3(13.7 mmol) over 1 hour, maintaining the pH at 6.5 by the addition of 73.5 ml of 1 M aqueous solution of meglumine (73,5 mmol). Exercise nanofiltration reaction mixture and bring the pH to 6.8 by adding 0.3 ml of 0.1 M meglumine. After evaporation and drying obtain the desired product (17,2 g; 12.9 mmol).

Spectra IR and MS correspond to the specified structure.

EXAMPLE 4

The gadolinium complex of 1-deoxy-1-(methylamino)-D-glaciology salt [3β (S), 5β , 12α ]-3-[[4-[bis[2-[bis(carboxymethyl)amino]ethyl]amino]-4-carboxy-1-oxobutyl]amino]-12-hydroxypoly-24-OIC acid (1:3)

A) Methyl ester [3β (S), 5β , 12α ]-3-[[4-[bis[2-[bis[2-(1,1-dimethylmethoxy)-2-oxoethyl]amino]ethyl]amino]-5-(1,1-dimethylmethoxy) -1, 5-dioxaphetyl] amino] -12-hydroxypoly-24-OIC acid

The triethylamine (2,23 g; 22 mmol) are added to a solution 8,93 g methyl ester (3β , 5β , 12β )-3-amino-12-hydroxypoly-24-OIC acid (obtained similarly derived the hall of the second acid, described in WO-A-95/32741: example 5) (22 mmol), 16,41 g of 1-(1,1-dimethylethanol) of ester N,N-bis[2-[bis[2-(1,1-dimethylmethoxy)-2-oxoethyl]amino]ethyl]-L-glutamato acid (obtained as described in WO-A-95/32741: example 15) (22 mmol) and 3,91 g diethylthiophosphate (24 mmol) in 300 ml of DMF at 0° C. After 1.5 hours at 0° and 18 hours at room temperature, the reaction mixture is evaporated and the residue dissolved in EtOAc. The solution was washed with saturated aqueous Panso3and H2O, dried (Na2SO4) and evaporated. The crude product is purified using flash chromatography, obtaining the desired product (20,67 g; 18.2 mmol).

Yield: 83%

[α ]

20
D
=-6,75 (with 2.0, l3)

Elemental analysisNN
% calculated:65,699,604,94
% found:66,549,954,99

TLC: Stationary phase: a plate of silica gel 60F 254 Merck

Eluent: 1:1 n-hexane/EtOAc Rf=0,09

Detection: CE(SO4)2·4H2O (0,18%) and (NH4)6Mo7O24·4H2O (3,83%) in 10% H2SO4

Spectra1 H-NMR,13C-NMR, IR and MS correspond to the specified structure.

In) [3β (S), 5β , 12α )-3-[[4-[bis[2-[bis(carboxymethyl)amino]ethyl]amino]-4-carboxy-1-oxobutyl]amino]-12-hydroxypoly-24-OIC acid

The compound obtained in stage A) (of 19.72 g; of 17.4 mmol), dissolved in 105 ml of CF3CO2H at room temperature. After 26 hours the solution is evaporated and the residue is treated with N2About; the hard part is filtered off, washed with N2Oh and partially dried in vacuum. The obtained intermediate compound suspended in N2Oh and dissolve 1 M NaOH to pH 13. After 5 hours at room temperature is added dropwise a 0.5 M Hcl solution until the pH of 1.4. After 15 hours at room temperature the precipitate is filtered off, washed with N2O and dried, give crude product which is purified chromatographically on a column of resin Amberlite® XAD 1600 with obtaining the desired product (9,92 g; and 11.8 mmol).

Yield: 68%. TPL: 184° C (decomposition)

Complexometrically titer (0.1 M GdCl3): 99,3%

Acid titer (0.1 n NaOH): 99,8%

[α ]20λ(2,0; 1 M NaOH)

+23,61
λ (nm)589578546436405365
[α ]+24,59+27,90+46,67+55,61+71,40
Elemental analysisNN 
% calculated:58,70to 7.93of 6.68 
% found:58,228,166,59H2About0,70%

TLC: Stationary phase: a plate of silica gel 60F 254 Merck

Eluent: 5:4:2 l3/Meon/25% NH4OH Rf=0,13

Detection: CE(SO4)2·4H2O (0,18%) and (NH4)6Mo7O24·4H2O (3,83%) in 10% H2SO4

Spectra1H-NMR,13C-NMR, IR and MS correspond to the specified structure.

C) gadolinium Complex of 1-deoxy-1-(methylamino)-D-glaciology salt [3β (S), 5β , 12α )-3-[[4-[bis[2-[bis(carboxymethyl)amino]ethyl]amino]-4-carboxy-1-oxobutyl]amino]-12-hydroxypoly-24-OIC acid (1:3)

The compound obtained in stage C) (8,39 g; 10 mmol), suspender the t in H 2O (30 ml) and dissolved by adding 1 M aqueous solution of meglumine (to 36.5 ml of 36.5 mmol) up to pH 6. To the mixture is added dropwise 1,025 M aqueous solution GdCl3(9,85 ml; 10.1 mmol) over 1 hour, maintaining the pH 6 by addition of 1 M aqueous solution of meglumine (19.3 ml; or 19.3 mmol). Carry out the nanofiltration of the solution and the pH was adjusted to 7.0 1 M aqueous solution of meglumine. After evaporation and drying obtain the desired product (to 8.57 g; 5.4 mmol).

Yield: 54%. TPL: 150-166° (170° With Razlog.)

Elemental analysisNNGd 
% calculated:47,177,286,219,96 
% found:43,407,315, 689,31H2O 7,14%

Spectra IR and MS correspond to the specified structure.

D) Similarly, the compound obtained in stage C), receive a complex of gadolinium with a sodium salt of [3β (S), 5β , 12α )-3-[[4-[bis[2-[bis(carboxymethyl)amino]ethyl]amino]-4-carboxy-1-oxobutyl]amino]-12-hydroxypoly-24-OIC acid (1:3)

The compound obtained in stage C) (26,92 g; 32,08 mmol), suspended in N2O (100 ml) and dissolved by adding 2 M NaOH (56 ml, 112 mmol) to pH 6. To the mixture until ablaut dropwise 0,512 M aqueous solution GdCl 3(58,2 ml; 29,77 mmol) for 3 hours, maintaining the pH 6 by adding 2 M NaOH (28,95 ml; to 57.9 mmol). Bring the solution pH to 6.7 by addition of 2 M NaOH (4 ml, 8 mmol) and carry out the nanofiltration of the solution. After freeze-drying obtain the desired product (29,86 g of 28.2 mmol).

Yield: 88%. TPL: >300°

Elemental analysisNNGdNa 
% calculated:46,495,71of 5.2914,856,51 
% found:43,986,344,9213,866,61H2About 4,63%

Spectra IR and MS correspond to the specified structure.

EXAMPLE 5

An alternative way to obtain [3β (S), 5β , 12α )-3-[[4-[bis[2-[bis(carboxymethyl)amino]ethyl]amino]-4-carboxy-1-oxobutyl]amino]-12-hydroxypoly-24-OIC acid according to scheme 3

A) 2-methyl-1-(fenilmetilovy) W (S)-5-oxo-1,2-pyrrolidinecarbonyl acid

7,1 g CH3I (50 mmol) are added to the solution to 6.58 g of 1-(phenylmethylene) ether (S)-5-oxo-1,2-pyrrolidinecarbonyl acid (25 mmol) and N,N-diisopropylethylamine (3.55 g; 27.5 mmol) in CH2Cl2(33 ml) and the reaction mixture is boiled OBR is Tim fridge for 6.5 hours. After cooling to room temperature and dilution CH2Cl2(50 ml), the reaction mixture was washed with H2Oh, 2% aqueous PA2CO3, 0.2 M Hcl and H2O. After drying over Na2SO4and evaporation to obtain the desired product (6.8 g; 24.5 mmol).

Yield: 98%

HPLC analysis: 98/5% (area %)

Spectra1H-NMR,13C-NMR, IR and MS correspond to the specified structure.

C) Methyl ester [3β (S), 5β , 12α ]-3-[[5-methoxy-1,5-dioxo-4-[[(phenylmethoxy)carbonyl]amino]pentyl]amino]-12-hydroxypoly-24-OIC acid

of 8.92 g methyl ester (3β , 5β , 12α )-3-amino-12-hydroxypoly-24-OIC acid (obtained similarly derived holeva acid, described in WO-A-95/32741: example 5) (22 mmol) are added to a solution of compound A) (6,1 g; 22 mmol) in dioxane (55 ml) and the resulting mixture was heated to 50° within 24 hours, then to 105° With over 29 hours. The solvent is evaporated under reduced pressure and the residue purified using flash chromatography (gradient elution EtOAc/n-hexane) followed by crystallization from a mixture of 1:1 EtOAc/n-hexane, obtaining the desired product (11.2 g; 16.4 mmol).

Yield: 75%. TPL: 140° C.

HPLC analysis: 99,2% (area %)

Stationary phase:Lichosorb RP-Select 5 μm; 250× 4 mm column of Merck KGaA
Temperature:45° C
Mobile phase:gradient elution
A=0,017 M H3RHO4in the water
In=CH3SP
Gradient:min%%
 06535
 251585
 301585

Flow rate: 1 ml/min

Detection (UV): 210 nm

[α ]20λ=(2,01; l3)

λ (nm)589578546365
[α ]+24,14+25,13+28,51+73,81
Elemental analysisNN
% calculated:68,708,434,11
% found:69,368,72 4,13

TLC: Stationary phase: a plate of silica gel 60F 254 Merck

Eluent: EtOAc

Detection: CE(SO4)2·4H2O (0.2%) and (NH4)6Mo7O24·4H2O (3,8%) in 10% H2SO4Rf=0,11

Spectra1H-NMR,13C-NMR, IR and MS correspond to the specified structure.

C) Methyl ester [3β (S), 5β , 12α ]-3-[[4-[bis[2-[bis[2-(1,1-dimethylmethoxy)-2-oxoethyl]amino]ethyl]amino]-5-methoxy-1,5-dioxaphetyl]amino]-12-hydroxypoly-24-OIC acid

1 g of 5% Pd/C are added to a solution of the compound In) (10.4 g; and 15.3 mmol) in Meon (100 ml); the suspension is stirred for 3.5 hours in an atmosphere of hydrogen absorbed in the N2: 348 ml of 15.5 mmol) at room temperature. After filtration through a Millipore filter® FH (0.45 µm) solution is evaporated under reduced pressure, obtaining a residue, which was dissolved in CH3JV (60 ml). Add 2 M of water (pH 8) phosphate buffer (60 ml), then a solution of 1,1-dimethylethylene ether N-(2-bromacil)-N-[2-(1,1-dimethylmethoxy)-2-oxoethyl]glycine (obtained by the method, opened in WO-A-95/32741: example 15) (up 11,86 g; 33,7 mmol) in CH3JV (15 ml) is added dropwise during 10 min at room temperature. The mixture is stirred for 39 hours. After separation the organic phase is evaporated under reduced pressure and the residue is dissolved in AcOEt (200 ml). The solution is washed with N2the, dried (Na2SO4) and evaporated. The crude product is purified using flash chromatography (gradient elution EtOAc/n-hexane)to give the desired product (11,36 g; 10.4 mmol).

Yield: 68%. TPL: 55-58°

HPLC analysis: 100% (area %)

[α ]20λ=(2,01; l3)

λ (nm)589578546365
[α ]-6,97-7,41-8,61-32,89
Elemental analysisNN
% calculated:64,939,425,13
% found:65,069,365,11

TLC: Stationary phase: a plate of silica gel 60F 254 Merck

Eluent: EtOAc

Detection: CE(SO4)2·4H2O (0.2%) and (NH4)6Mo7O24·4H2O (3,8%) in 10% H2SO4Rf=0,45

Spectra1H-NMR,13C-NMR, IR and MS correspond to the specified structure.

D) [3β (S), 5β , 12α)-3-[[4-[bis[2-[bis(carboxymethyl)amino]ethyl]amino]-4-carboxy-1-oxobutyl]amino]-12-hydroxypoly-24-OIC acid

To a solution of compound (8.5 g; 7.8 mmol) in dioxane (50 ml) is added 2 M aqueous solution of LiOH (117 ml, 234 mmol). The resulting mixture was stirred at room temperature for 72 hours, then acidified to pH 6 by slowly adding 37% Hcl. The solution is concentrated to 50 g, pariva under reduced pressure, and diluted with N2O (40 ml). The solution is acidified to a pH of 2.5 by the addition of 37% Hcl, heated to 50-55° and under intensive stirring very slowly acidified to a pH of 1.3, adding 2 N. Hcl. After 5 min heterogeneous mixture is left for slow cooling to room temperature with stirring for 15 hours. The precipitate is filtered off, washed with H2O and dried, obtaining the desired product (of 5.92 g; 7 mmol).

Yield: 90%. TPL: 180-198° C.

HPLC analysis: 99,9% (area %)

Spectra1H-NMR,13C-NMR, IR and MS correspond to the specified structure.

EXAMPLE 6

The gadolinium complex of 1-deoxy-1-(methylamino)-D-glaciology salt (3β , 5β , 7α , 12α )-3-[[[[[bis[2-[bis(carboxymethyl)amino]ethyl]amino]acetyl]amino]acetyl]amino]-7,12-dihydroxyfuran-24-OIC acid (1:2)

A) N-[[bis[2-[bis[2-(1,1-dimethylmethoxy)-2-oxoethyl]amino]ethyl]amino]acetyl]glycine

6.5 g of glycylglycine (to 49.3 mmol) are suspended in 100 ml of a mixture 1:1 H2O/EtOH and dissolved at pH 10 by adding 10 M NaO (4.8 ml). To this is added dropwise 1,1-dimethylethylene ester of N-(2-bromacil)-N-[2-(1,1-dimethylmethoxy)-2-oxoethyl]glycine (42 g; 110,9 mmol) in 40 ml EtOH for 2 hours, maintaining the pH at 10.5 by addition of 10 M NaOH (5.8 ml). The solution is rapidly converted into an emulsion, which is dissolved in 2.5 hours by adding 10 M NaOH. After 22 hours the solvent is evaporated, the mixture is diluted with water and extracted with CH2CL2. The organic phase is washed with N2Oh, dried and evaporated, receiving the remnant that purify using flash chromatography. The residue is dissolved in water, adjusted pH to 4.5 by addition of 1 M Hcl and the solution extracted with chloroform. The organic phase is washed with N2O, dried and evaporated, obtaining 13 g of the desired product (to 19.3 mmol).

Yield: 39%

Elemental analysisNN
% calculated:56,958, 668,30
% found:56, 678, 688,30

TLC: Stationary phase: a plate of silica gel 60F 254 Merck

Eluent: 6:3:1 l3/Meon/25% NH4OH

Rf=0,65

Detection: 1% KMnO4in 1 M NaOH

Spectra1H-NMR,13C-NMR, IR and MS correspond to the specified structure.

C) Methyl ether (3β , 5β , 7α , 12α )-3-[[[[[bis[2-[bis[2-(11-dimethylmethoxy)-2-oxoethyl]amino]ethyl]amino]acetyl]amino]acetyl]amino]-7,12-dihydroxyfuran-24-OIC acid

2,8 ml TEA (triethanolamine) (a 20.2 mmol) is added dropwise over 5 min to a solution containing of 13.6 g of compound obtained in stage A) (a 20.2 mmol), charged 8.52 g methyl ester (3β , 5β , 7α , 12α )-3-amino-7,12-dihydroxyfuran-24-OIC acid (20,2 mmol) and DEPC (3.4 ml; of 22.2 mmol) in DMF (290 ml), stir at 0° C. After 1 hour the reaction mixture is heated to room temperature and the solution stirred for 6.5 hours. Add 0.3 ml of DEPC (2 mmol) and the solution stirred for additional 15.5 hours. DMF is evaporated, the residue is dissolved in EtOAc, washed with water Panso3and finally dried. After purification using flash chromatography gain of 13.7 g of the desired product (12.7 mmol).

Yield: 63%.

[α ]20D=+5,26 (1.5; l3)

Elemental analysisNN
% calculated:63,48a 9.25of 6.49
% found:63,229,406,40

Spectra1H-NMR,13C-NMR, IR and MS correspond to the specified structure.

(C) (3β , 5β , 7α , 12α )-3-[[[[[bis[2-[bis(carboxymethyl)amino]ethyl]amino]acetyl]amino]acetyl]amino]-7,12-dihydroxyfuran-24-OIC acid

is 12.85 g of compound obtained in stage C) (12 mmol), dissolved in TFU (210 ml), paramashiva the Oh-5/0° C. After 16 hours TFU is evaporated, obtaining a residue which is dissolved in 90 ml of 0.8 M NaOH at pH 13 and stirred at room temperature for 15 hours. The solution is concentrated to 50 ml, was added dropwise in 105 ml of 0.6 M Hcl and stirred for 2 hours. The hard part is filtered off, washed with 0.1 M Hcl and dried, obtaining the crude product, which was purified using flash chromatography. Fractions containing the desired compound in the form of a salt, evaporated, obtaining a residue which is dissolved in water, and added dropwise in 1 M Hcl, while maintaining the pH of 1.45. The precipitate is filtered off, washed with 0.1 M Hcl and dried, obtaining 2.6 g of the desired product (3.1 mmol).

Yield: 26%. TPL: 120-125° C.

HPLC assay: 98% (area %)

Spectra1H-NMR,13C-NMR, IR and MS correspond to the specified structure.

D) gadolinium Complex of 1-deoxy-1-(methylamino)-D-glaciology salt (3β , 5β , 7α , 12α )-3-[[[[[bis[2-[bis[2-(carboxymethyl)amino]ethyl]amino]acetyl]amino]acetyl]amino]-7,12-dihydroxyfuran-24-OIC acid (1:2)

2,59 g of compound obtained in stage C) (is 3.08 mmol), suspended in water (20 ml) and dissolved by adding 1 M aqueous solution of meglumine (is 3.08 ml, is 3.08 mmol) to pH 5. To this mixture Gd2O3(0,501 g; 2.77 mmol), heating to 50° C. After 1 hour, add 1 M meglumin (2,8 ml; 2.8 mmol) to dissolve the precipitate. After 24 hours the reaction mixture is filtered and adjusted the pH to 6,dobavleniem 1 M aqueous meglumine (0.4 ml). After evaporation and drying gain of 4.2 g (3.00 mmol) of the desired product.

Yield: 99%. TPL: 209-213° C (decomposition)

HPLC analysis: 99,7% (area %)

The free ligand:<0,1%(0,001 GdCl3)
Elemental analysisHNGd 
% calculated:46,846,997,0811,36 
% found:44,017,35of 6.68accounted for 10.39H2O 4,95

Spectra IR and MS correspond to the specified structure.

EXAMPLE 7

The gadolinium complex of 1-deoxy-1-(methylamino)-D-glaciology salt [3β (S), 5β ]-3-[[4-[bis[2-[bis(carboxymethyl)amino]ethyl]amino]-4-carboxy-1-oxobutyl](carboxymethyl)amino]Holan-24-OIC acid (1:4)

A) Methyl ether (3β , 5β )-3-[[2-(1,1-dimethylmethoxy)-2-oxoethyl]amino]Holan-24-OIC acid

40 g methyl ester (3β , 5β )-3-Linkoln-24-OIC acid (obtained in example 1) (103 mmol) suspended in DMF (1.0 l) at room temperature under nitrogen atmosphere. Add triethylamine (13,0 g, 129 mmol), then the reaction mixture is added dropwise Rast is the PR 1,1-dimethylethylene ether bromoxynil acid (24,0 g; 123 mmol) in DMF (30 ml) for 1 hour to dissolve. After 3 days the mixture was concentrated and diluted with 4% aqueous solution Panso3. The resulting suspension is filtered, the residue washed with N2O and dried, obtaining 33,7 g (66,9 mmol) of the desired product.

Yield: 65%. TPL: 62-64°

[α ]20D=+23,55 (from 1.96, Meon)

Elemental analysisNN 
% calculated:73,91or 10.602,78 
% found:74,6710,852,78H2O<0,1

TLC: Stationary phase: a plate of silica gel 60F 254 Merck

Eluent: 7:3 n-hexane/EtOAc Rf=0,31

Spectra1H-NMR,13C-NMR, IR and MS correspond to the specified structure.

C) Methyl ester [3β (S), 5β ]-3-[[4-[bis[2-[bis[2-(1,1-dimethylmethoxy)-2-oxoethyl]amino]ethyl]amino]-5-(1,1-dimethylmethoxy)-1,5-dioxaphetyl][2-(1,1-dimethylmethoxy)-2-oxoethyl]amino]Holan-24-OIC acid

Diisopropylethylamine (19.5 g; 151 mmol) is added dropwise over 20 minutes to a solution of compound A) (33,0 g; 65,5 mmol), 1-(1,1-dimethylethanol) ether N,N-bis[2-[bis[2-(1,1-dimethylmethoxy)-2-oxoethyl]amino]ethyl]-L-glutamato acid (obtained as described in WO-A-95/32741: example 15) (53,8 g; 72,1 mmol) and sexafter state (benzotriazol-1 yloxy)Tris(dimethylamino)phosphonium (THIEF) (40.6 g; about 91.8 mmol) in DMF (400 ml), stir at room temperature under nitrogen atmosphere. After 2 days the reaction mixture was concentrated and absorb EtOAc. The solution is washed with N2O, dried (Na2SO4and viparitakarani product twice purified using flash chromatography, obtaining the desired product (38,7 g; of 31.4 mmol).

Yield: 48%

[α ]20D=-54,50 (of 2.51, l3)

TLC: Stationary phase: a plate of silica gel 60F 254 Merck

Eluent: 7:3 n-hexane/EtOAc Rf=0,22

Spectra1H-NMR,13C-NMR, IR and MS correspond to the specified structure.

C) [3β (S), 5β ]-3-[[4-[bis[2-[bis(carboxymethyl)amino]ethyl]amino]-4-carboxy-1-oxobutyl](carboxymethyl)amino]Holan-24-OIC acid

Into a solution of the compound obtained in stage C) (38,7 g; of 31.4 mmol)in EtOH (350 ml), stir at room temperature, and added dropwise to 500 ml of 2 M solution of LiOH in 1 hour. After 16 hours the reaction mixture was concentrated to 500 ml and again add 2 M LiOH solution (350 ml), heating to 50° C. After 24 hours the reaction mixture is cooled to room temperature and added dropwise in 320 ml of 6 M Hcl, intensively stirring at 5° C. the pH of the resulting suspension was adjusted to 1.0 by adding 55 ml of 2 M NaOH. The hard part is filtered off, washed with 0.1 M Hcl and dried. The crude product is suspended in N2Oh and solubilizers by adding 1 M NaOH solution, then the basis of the Noi solution is added dropwise in 0.5 M Hcl solution. The precipitate is filtered off, washed with 0.05 M Hcl, H2O and dried, obtaining of 23.5 g of the desired product (to 26.7 mmol).

Yield: 85%. TPL: 178-182°

[α ]

405
20
=+24,10 (1,49, 1 M NaOH)

HPLC analysis: 100% (area %)

Stationary phase:Hypurity Elite C-18, 5 μm; column 250× 4.6 mm
Temperature:40°
Mobile phase:gradient elution
 A=0.01 M KN2RHO4, 0.3 mm EDTA in water
 In=CH3SP
Gradient:min%%
 0955
 406535
 506535

Flow rate: 1 ml/min

Detection (UV): 210 nm

Ale is ntny analysis NNNaCL 
% calculated:58,627,786,36   
% found:57,718,076,200,130,55H2O<0,1

Spectra1H-NMR,13C-NMR, IR and MS correspond to the specified structure.

D) gadolinium Complex of 1-deoxy-1-(methylamino)-D-glaciology salt [3β (S), 5β ]-3-[[4-[bis[2-[bis(carboxymethyl)amino]ethyl]amino]-4-carboxy-1-oxobutyl](carboxymethyl)amino]Holan-24-OIC acid (1:4)

To a suspension of the compound obtained in stage C) (12.9 g; 14.6 mmol), in N2O (100 ml) was added 1 M aqueous solution of meglumine (72,0 ml, 72.0 mmol) at room temperature to obtain a clear solution with a pH of 6.2. To the mixture is added dropwise 0,393 M aqueous solution GdCl3(37,2 ml, 14.6 mmol), maintaining the pH of 6.2 by addition of 1 M aqueous solution of meglumine (30,0 ml; 30.0 mmol) using a pH-stat. At the end of the additions the reaction mixture is filtered through paper, and then through the membrane Millipore® (HAWP 0.45 µm), carry out the nanofiltration of the solution was adjusted pH to 7.0 by adding 1 M aqueous solution of meglumine (of 0.20 ml, 0.20 mmol), and evaporated. The hard part is dried, obtaining the desired product (24,0 g; 13,mmol).

Yield: 91%. TPL: 90-92°

Spectra IR and MS correspond to the specified structure.

EXAMPLE 8

A complex of gadolinium with a sodium salt of [3β (R), 5β , 12α ]-3-[[4-[bis[2-[bis(carboxymethyl)amino]ethyl]amino]-4-carboxy-1-oxobutyl]amino]-12-hydroxypoly-24-OIC acid (1:3)

In accordance with scheme 3 and the experimental method of example 5 1-(fenilmetilovy) ether (R)-5-oxo-1,2-pyrrolidinecarbonyl acid (commercially available product) esterification methyliodide in the presence of N,N-diisopropylethylamine and thus obtained (R) methyl ether is subjected to interaction with the methyl ether (3β , 5β , 12α )-3-amino-12-hydroxypoly-24-OIC acid (obtained similarly derived holeva acid, described in WO-A-95/32741: example 5)to give methyl ester [3β (R), 5β , 12α ]-3-[[4-[bis[2-[bis[2-(1,1-dimethylmethoxy)-2-oxoethyl]amino]ethyl]amino]-5-methoxy-1,5-dioxaphetyl]amino]-12-hydroxypoly-24-OIC acid. After removal of the (H2/Pd) Cbz protective group and alkylation of 1,1-dimethylethylene ether N-(2-bromacil)-N-[2-(1,1-dimethylmethoxy)-2-oxoethyl]glycine (obtained by the method, opened in WO-A-95/32741: example 15) in CH3CN/phosphate buffer pH 8, get hexa (ester), which make (aqueous LiOH/dioxane) to the appropriate hexakis is at. Form a complex of the latter by the method of example 4, paragraph (D), resulting in getting the right product with an overall yield of 44%.

TPL: >300°

Elemental analysisHNGdNa
% calculated:46,495,71of 5.2914,856,51
% found:44,026,135,1014,096,17
 H2O 4,50%

Spectra IR and MS correspond to the specified structure.

EXAMPLE 9

A complex of gadolinium with a sodium salt of [3β (RS), 5β , 12α ]-3-[[4-[bis[2-[bis(carboxymethyl)amino]ethyl]amino]-4-carboxy-1-oxobutyl]amino]-12-hydroxypoly-24-OIC acid (1:3)

The connection is obtained from the methyl ester (3β , 5β , 12α )-3-amino-12-hydroxypoly-24-OIC acid (obtained similarly derived holeva acid, described in WO-A-95/32741: example 5)and 1-(1,1-dimethylethanol) ether N,N-bis[2-[bis[2-(1,1-dimethylmethoxy)-2-oxoethyl]amino]ethyl]-DL-glutaminol acid (obtained from DL-glutaminol acid, as described in WO-A-95/32741: example 15 L-isomer) in accordance with the method detailed in Primera. The product is obtained with a total yield of 61%.

TPL: >300°

HPLC analysis: 99% (area %)

Stationary phase:Bond ECLIPSE XDB-C8 3.5 µm; 150× 4.6 mm column Rockland Technologies, Inc.
Temperature:40° C
Mobile phase:gradient elution
 A=0,005 M KH2PO4, 0.005 M2NRA4for 0.3
 mm EDTA in water
 In=CH3CN
Gradient:min%%
 09010
 59010
 205050

Flow rate: 1.0 ml/min

Detection (UV): 210 nm

Chromatographic method reveals two peaks, almost equal in percentage terms that are associated with diastereoisomers due to the presence of RS stereocenter in DTPA residue.

Elemental analysisNNGdNa
% calculated:46,495,71of 5.2914,856,51
% found:43,986,34to 4.9813,866,16
 H2O 4,63%    

Spectra IR and MS correspond to the specified structure.

EXAMPLE 10

A complex of gadolinium with a sodium salt of [3α (S), 5β , 12α ]-3-[[4-[bis[2-[bis(carboxymethyl)amino]ethyl]amino]-4-carboxy-1-oxobutyl]amino]-12-hydroxypoly-24-OIC acid (1:3)

Methyl ether (3α , 5β , 12α )-3-amino-12-hydroxypoly-24-OIC acid

Methyl ether (3α , 5β , 12α )-3,12-dihydroxyfuran-24-OIC acid is subjected to interaction in the Mitsunobu conditions (Mitsunobu, O. Synthesis 1981, 1-28; Denike, J.K. et al. Chem. Phys. Lipids 1995, 77, 261-267) with triphenylphosphine, diethylazodicarboxylate and formic acid in THF, receiving methyl ester (3β , 5β , 12β )-3-formyloxy-12-hydroxypoly-24-OIC acid. Remove the protective group (MeOH/HCl) to the methyl ester (3β , 5β , 12α )-3,12-dihydroxyfuran-24-OIC acid, which is injected in a series of reactions is, described in WO-A-95/32741: example 5) to obtain the derived holeva acid. Methyl ether (3α , 5β , 12α )-3-amino-12-hydroxypoly-24-OIC acid get with a total yield of 32%.

TPL: 90-92°

[α ]20D=+53,46 (1,3, CH3IT)

Elemental analysisNN
% calculated:74,0310,693,44
% found:74,2010,993,26

TLC: Stationary phase: a plate of silica gel 60F 254 Merck

Eluent: 9:1:0,15 l3/CH3HE/25% NH4OH Rf=0,21

Spectra1H-NMR,13C-NMR, IR and MS correspond to the specified structure.

In accordance with reaction scheme 3 and the experimental method presented in example 5, 2-methyl-1-(fenilmetilovy) W (S)-5-oxo-1,2-pyrrolidinecarbonyl acid is subjected to interaction with the compound (A). Thus obtained methyl ester [3α (S), 5β , 12α ]-3-[[4-[bis[2-[bis[2-(1,1-dimethylmethoxy)-2-oxoethyl]amino]ethyl]amino]-5-methoxy-1,5-dioxaphetyl]amino]-12-hydroxypoly-24-OIC acid hydronaut (H2/PD) to remove the Cbz protective group. Subsequent alkylation of 1,1-dimethylethylene ether N-(2-bromacil)-N-[2-(1,1-dimethylmethoxy)-2-oxoethyl]glycine (the floor is built according to the method, opened in WO-A-95/32741: example 15) in CH3SP/phosphate buffer pH 8, leads to the production of hexa(of ester). The last turn (aqueous LiOH/dioxane) in hexacyclic, which is completed by the method of example 4, paragraph (D), getting the right product with the full output of 33%.

TPL: >300° C.

HPLC analysis of 100% (area %)

Elemental analysisNNGdNa
% calculated:46,495,71of 5.2914,856,51
% found:42,046,374,7613,285,91
 H2O 9,79%    

Spectra IR and MS correspond to the specified structure.

EXAMPLE 11

A complex of gadolinium with a sodium salt of [3β (S), 5β , 7α ]-3-[[4-[bis[2-[bis(carboxymethyl)amino]ethyl]amino]-4-carboxy-1-oxobutyl]amino]-7-hydroxypoly-24-OIC acid (1:3)

The connection is obtained from the methyl ester (3β , 5β , 7α )-3-amino-7-hydroxypoly-24-OIC acid (obtained similarly derived holeva acid, described in WO-A-95/32741: example 5) and 1-(1,1-dimethyle is gross) ether N,N-bis[2-[bis[2-(1,1-dimethylmethoxy)-2-oxoethyl]amino]ethyl]-L-glutamato acid (obtained as described in WO-A-95/32741: example 15) according to the method, described in detail in example 4. The product is obtained with a total of 89% yield.

TPL: >300°

Elemental analysisNNGdNa
% calculated:46,495,71of 5.2914,856,51
% found:43,886,504,9113,626,04
 H2About 7,11%    

Spectra IR and MS correspond to the specified structure.

EXAMPLE 12

[3β (RS), 5β , 7α , 12α ]-3-[[4-[bis[2-[bis(carboxymethyl)amino]ethyl] amino]-4-carboxy-1-oxobutyl]amino]-7,12-dihydroxyfuran-24-OIC acid

The connection is obtained from the methyl ester (3β , 5β , 7α , 12α ]-3-amino-7,12-dihydroxyfuran-24-OIC acid (obtained as described in WO-A-95/32741: example 5) and 1-(1,1-dimethylethanol) ether N,N-bis[2-[bis[2-(1,1-dimethylmethoxy)-2-oxoethyl]amino]ethyl]-DL-glutaminol acid (obtained from DL-glutaminol acid, as described in WO-A-95/32741: example 15 L-isomer) according to the method detailed in example 4 to obtain a connection). The product is obtained with a total of 65% yield./p>

TPL: 224° C (decomposition)

HPLC analysis: 97% (area %)

Stationary phase: Bond ECLIPSE XDB-C8 3.5 µm; 150× 4.6 mm column Rockland Technologies, Inc.

Temperature: 40°

Mobile phase: gradient elution

A=0,017 M H3RHO4, 0.3 mm EDTA in water

In=CH3JV

Gradient:min%%
 0955
 406535
 506535

Flow rate: 1.0 ml/min

Detection (UV): 210 nm

Chromatographic method reveals two peaks, in almost equal percentages that are associated with diastereoisomers due to the presence of RS-stereocenter in DTPA residue.

Elemental analysisNNLi, Cl 
% calculated:57,607,786,55  
% found:54,347,82to 6.19<0,1H2O 5,12%

Spectra of IR, NMR and MS match the specified structure is r.e.

EXAMPLE 13

A complex of gadolinium with a sodium salt of [3α (S), 5β , 7α , 12α ]-3-[[[[5-[bis[2-[bis(carboxymethyl)amino]ethyl]amino]-5-carboxypentyl]amino]carbonyl]oxy]-7,12-dihydroxyfuran-24-OIC acid (1:3)

A) Methyl ester [3α (S), 5β , 7α , 12α ]-3-[[[[6-(1,1-dimethylmethoxy)-5-[bis[2-[bis(2-(1,1-dimethylmethoxy)-2-oxoethyl]amino]ethyl]amino]-6-oxohexyl]amino]carbonyl]oxy]-7,12-dihydroxyfuran-24-OIC acid

Method 1: a Solution of bis(trichloromethyl)carbonate (2.9 g; 9.7 mmol) in anhydrous CH2Cl2(40 ml) is added dropwise under nitrogen atmosphere to a solution of methyl ester (3α , 5β , 7α , 12α )-3,7,12-trihydroxypregn-24-OIC acid (commercial product) (10.0 g; of 23.7 mmol) and pyridine (2.3 ml; 28.4 mmol) in anhydrous CH2CL2(100 ml), cooled to 0° C. the Mixture is left to warm up and after 1 hour at room temperature the solution is cooled again to 0° add N,N-diisopropylethylamine (7.9 ml; of 47.3 mmol), then added dropwise a solution of (1,1-dimethylethanol) ether N2N2bis[2-[bis[2-(1,1-dimethylmethoxy)-2-oxoethyl]amino]ethyl]-L-lysine (Anelli, P.L. et al. Bioconjugate Chem. 1999, 10, 137) (17.6 g; of 23.7 mmol) in anhydrous CH2CL2(50 ml). The solution is stirred for 3 hours at room temperature, then washed with N2About (2× 100 ml), dried over Na2SO4and evaporated. what ever got the product was then purified using flash chromatography getting the right product (14.8 g; 12.4 mmol).

Yield: 52%.

Spectra1H-NMR,13C-NMR, IR and MS correspond to the specified structure.

Method 2: a Solution of methyl ester [3α , 5β , 7α , 12α ]-3-[(chlorocarbonyl)oxy]-7,12-dihydroxyfuran-24-OIC acid (Janout, V., Lanier, M.; Regen, S. L. J. Am. Chem. Soc. 1997, 119, 640) (6,1 g; 12.6 mmol) in anhydrous CH2Cl2(150 ml) cooled to 0°C in an atmosphere of nitrogen and then added N,N-diisopropylethylamine (4.8 ml; 27.6 mmol). Then add dropwise a solution of (1,1-dimethylethanol) ether N2N2bis [2-[bis[2-(1, 1 dimethylmethoxy) -2-oxoethyl]amino] ethyl]-L-lysine (9.3 g; 12.6 mmol) in anhydrous CH2Cl2(20 ml). The solution is stirred for 3 hours at room temperature, then washed with N2About (2× 100 ml), dried over Na2SO4and evaporated. The crude product is purified using flash chromatography, obtaining the desired product (11.9 g; 9.9 mmol).

Yield: 79%.

Spectra1H-NMR,13C-NMR, IR and MS correspond to the specified structure.

In) [3α (S), 5β , 7α , 12α ]-3-[[[[5-[bis[2-[bis(carboxymethyl)amino]ethyl]amino]-5-carboxypentyl]amino]carbonyl]oxy]-7,12-dihydroxyfuran-24-OIC acid

2 M Aqueous solution of LiOH (141 ml) is added dropwise to a solution of compound (A) (11.2 g; 9.4 mmol) in 1,4-dioxane (141 ml) at room temperature. Over 104 hours, the solution concentrate (150 ml) and added drop is 2 M aqueous solution of Hcl (175 ml): final pH value of 1.9. The precipitate is filtered off, washed with N2About (5× 50 ml) and dried in vacuum. The crude product is purified using flash chromatography. The hard part is dissolved in 10% water. CH3CN and pH adjusted to 1 by adding concentrated Hcl, then the solution is injected into the column with resin Amberlite® XAD 1600 (250 ml) and elute with a gradient of CH3SP/N2O. Contains the product fraction is evaporated, obtaining the desired product (4.3 g; 4.8 mmol).

Yield: 51%. TPL: 184-191°

K.F.: 4,36%

[α ]

20
D
=+19,5 (1, 1 M NaOH)

HPLC analysis: 97,3% (% area)

Spectra1H-NMR,13C-NMR, IR and MS correspond to the specified structure.

C) gadolinium Complex with the sodium salt of [3α (S), 5β , 7α , 12α ]-3-[[[[5-[bis[2-[bis(carboxymethyl)amino]ethyl]amino]-5-carboxypentyl]amino]carbonyl]oxy]-7,12-dihydroxyfuran-24-OIC acid (1:3)

The compound obtained in stage C) (4.7 g; 5.2 mmol), suspended in N2O (100 ml) and dissolved by adding 1 M NaOH (10 ml) to pH 6.5. Add a solution of GdCl3(1,9 g; 5.2 mmol) in N2O (17 ml), maintaining the pH value of 6.5 by adding 1 M NaOH (15.6 ml). After 1 hour at room temperature the solution is injected into the column with resin Amberlite® XAD 16.00 (250 ml), elwira gradient of CH3SP/H 2O. Contains the product fraction is evaporated, obtaining the desired product (4,2 g; 3.8 mmol).

Yield 72%. TPL: >300°

K.F.: 9,49%

[α ]

20
D
=+2,63 (2, N2O)

HPLC analysis: 100% (% area) (in the same way that on stage)

Spectra1H-NMR,13C-NMR, IR and MS correspond to the specified structure.

EXAMPLE 14

The complex manganese sodium salt [3β (S), 5β , 12α ]-3-[[4-[[2-[bis(carboxymethyl)amino]ethyl](carboxymethyl)amino]-4-carboxy-1-oxobutyl]amino]-12-hydroxypoly-24-OIC acid (1:3)

A) Methyl ester [3β (S), 5β , 12α ]-3-[[4-amino-5-methoxy-1,5-dioxaphetyl]amino]-12-hydroxypoly-24-OIC acid

3.4 g of 5% Pd/C are added to a solution of methyl ether [3β (S), 5β , 12α ]-3-[[5-methoxy-1,5-dioxo-4-[[(phenylmethoxy)carbonyl]amino]pentyl]amino]-12-hydroxypoly-24-OIC acid (example 5, the product C) (34,14 g; 50 mmol) in Meon (340 ml). The suspension is stirred at room temperature over 3.5 hours in an atmosphere of hydrogen. After filtration through a Millipore filter® FH (0.45 µm) solution is evaporated to dryness, obtaining the desired product (27,1 g; to 49.3 mmol).

Yield: 98.6%of the

Weight loss (50°; high vacuum):<0,1%

Elemental analysisNN
% calculated:67,85of 9.555,10
% found:68,589,725,12

[α ]20D=+36,61 (2,01, l3)

Acid titer (0.1 M Hcl): 94,6%

Spectra1H-NMR,13C-NMR, IR and MS correspond to the specified structure.

C) Methyl ester [3β (S), 5β , 12α ]-3-[[4-[[2-[bis[2-(1,1-dimethylmethoxy)-2-oxoethyl]amino]ethyl][2-(1,1-dimethylmethoxy)-2-oxoethyl]amino]-5-methoxy-1,5-dioxaphetyl]amino]-12-hydroxypoly-24-OIC acid

The compound obtained in stage A) (16.0 g; of 29.1 mmol), and 1,1-dimethylethylene ester of N-(2-bromacil)-N-[2-(1,1-dimethylmethoxy)-2-oxoethyl]glycine (13.3 g; 37.8 mmol) (obtained according to the method, opened in WO-A-95/32741: example 15) was dissolved in EtOAc (120 ml). After adding 2 M phosphate buffer pH 8 (120 ml) the mixture is intensively stirred for 2 hours, then the aqueous phase is replaced with fresh 2 M phosphate buffer pH 8 (120 ml) and stirred for 70 hours. The mixture is heated to 40° C for 12 hours, cooled to room temperature, separated and the organic phase is evaporated, obtaining a residue, which was dissolved in CH2Cl2(150 ml), washed with water (2× 100 ml), dried over Na2SO4and evaporated. The crude product is purified using flash XP is matography, getting the right product (12.3 g; 15.0 mmol).

Yield: 52%.

Elemental analysisNN
% calculated:65,909,465,12
% found:65,999,725,03

TLC: Stationary phase: a plate of silica gel 60F 254 Merck

Eluent: 1:1=EtOAc/n-hexane Rf=0,40

Detection: 1% KMPO4in 1 M NaOH

Spectra1H-NMR,13C-NMR, IR and MS correspond to the specified structure.

C) Methyl ester [3β (S), 5β , 12α ]-3-[[4-[[2-[bis[2-(1,1-dimethylmethoxy)-2-oxoethyl]amino]ethyl][2-(1,1-dimethylmethoxy)-2-oxoethyl]amino]-5-methoxy-1,5-dioxaphetyl]amino]-12-hydroxypoly-24-OIC acid

tert-Butylbromide (3,9 g; 20,1 mmol) is added dropwise to a solution of compound obtained in stage C) (11,0 g; a 13.4 mmol), and N,N-diisopropylethylamine (3.5 ml; of 20.1 mmol) in CH3The JV. The mixture is stirred at room temperature for 24 hours and then add N,N-diisopropylethylamine (0.9 ml; 4.0 mmol) and tert-butylbromide (0.8 g; 4.0 mmol). The mixture is stirred for 70 hours, separated and the organic phase is evaporated, obtaining a residue, which was dissolved in CH2Cl2(150 ml), washed with water (2× 100 ml), dried over Na2SO4and evaporated. The crude product eyes is try using flash chromatography getting the right product (10.7 g; 11.5 mmol).

Yield: 85%.

Elemental analysisNN
% calculated:65,579,394,50
% found:66,279,62to 4.52

TLC: Stationary phase: a plate of silica gel 60F 254 Merck

Eluent: 1:1=EtOAc/n-hexane Rf=0,46

Detection: 1% KMPO4in 1 M NaOH

Spectra1H-NMR,13C-NMR, IR and MS correspond to the specified structure.

D) [3β (S), 5β , 12α ]-3-[[4-[[2-[bis(carboxymethyl)amino]ethyl](carboxymethyl)amino]-4-carboxy-1-oxobutyl]amino]-12-hydroxypoly-24-OIC acid

2 M aqueous LiOH solution (120 ml) is added dropwise to a solution of compound obtained in stage C), (9,2 g; 9.8 mmol)in 1,4-dioxane (120 ml) at room temperature. After 24 hours the solution is concentrated (100 ml) and added dropwise to a 2 M aqueous Hcl (135 ml): final pH value of 1.9. The precipitate is filtered off, washed with N2About (5× 50 ml) and dried in vacuum, obtaining the desired product (7,3 g; 9.6 mmol).

Yield: 98%. TPL: 163-168°

K.F.: 1,81%

[α ]

20
D
=+29,03 (1, 1 M NaOH)

HPLC analysis: 100% (area %)

podvigina phase: Bond Eclipse XDB-C8 3.5 µm; 150× 4.6 mm; column filled Rockland Technologies Inc.;

Temperature: 40° C;

Spectra1H-NMR,13C-NMR, IR and MS correspond to the specified structure.

(E) a Complex of manganese sodium salt [3β (S), 5β , 12α ]-3-[[4-[[2-[bis(carboxymethyl)amino]ethyl](carboxymethyl)amino]-4-carboxy-1-oxobutyl]amino]-12-hydroxypoly-24-OIC acid (1:3)

The compound obtained in stage D) (5,3 g; 7.2 mmol), suspended in N2O (250 ml) and dissolved by adding 1 M NaOH (36 ml) to pH 6.5. Add a solution of MnCl2·4H2O (1.4 g; 7.2 mmol) in N2About (50 ml), maintaining the pH at 6.5 by adding 1 M NaOH (7.9 ml). After 1 hour at room temperature the solution absoluut by nanofiltration, then evaporated, getting the right product.

Yield: 98%. TPL: >300°

K.F.: 13,54%

[α ]20D=+2,63 (2, H2O)

CE analysis: 100%(area %)

Capillary: quartz glass 0,72 m × 50 µm

Voltage: 30 kV

Buffer: 0,07 M borate, pH of 9.3, 0.3 mm EDTA

Temperature: 25°

Stop time: 20 min

Detection (UV): 200 nm

Injection: hydrodynamic (50 mbar, 4 sec)

Concentration of sample: 1 mg/ml

Instrument: Hewlett Packard 3D NRSE

Temporary scheme prior training
t (min) action
2rinsing H2O
2rinsing with 0.1 M NaOH
1wash N2About
5flushing the buffer

Elemental analysisNNMnNaCL
% calculated:51,876,354,906,418,05 
% found:45,506,954,305,286,86<0,1

Spectra IR and MS correspond to the specified structure.

In the same way, on the basis of connections) of example 2, receive gadolinium complex [3β (S), 5β ]-3-[[4-[[2-[[bis(carboxymethyl)amino]ethyl](carboxymethyl)amino]-4-carboxy-1-oxobutyl]amino]-12-Oklahoman-24-OIC acid.

EXAMPLE 15

The complex manganese sodium salt [3α (S), 5β , 12α ]-3-[[[[5-[[2-[bis(carboxymethyl)amino]ethyl](carboxymethyl)amino]-5-carboxypentyl]amino]carbonyl]oxy]-12-hydroxypoly-24-OIC acid (1:3)

A) Methyl ester of N2-[2-[bis[2-(1,1-dimethylmethoxy)-2-oxoethyl]amino]ethyl]-N2-[2-(1,1-dimethylmethoxy)-2-oxoethyl]-N -[(phenylmethoxy)carbonyl]-L-lysine

1,1-dimethylethylene ester of N-(2-bromacil)-N-[2-(1,1-dimethylmethoxy)-2-oxoethyl]glycine (25.6 g; 72,55 mmol) (obtained according to the method, opened in WO-A-95/32741: example 15), dissolved in CH3JV (25 ml)is added dropwise to a solution of methyl ester hydrochloride N6-[(phenylmethoxy) carbonyl]-L-lysine (20 g; 60,46 mmol) and N,N-diisopropylethylamine (12,64 ml; 72,55 mmol) in CH3SP (250 ml). The reaction mixture was stirred at room temperature. 5 days to the solution add N,N-diisopropylethylamine (17,7 ml; 101,6 mmol) and tert-butylbromide (18,8 g; 13,5 ml; 101,6 mmol). After 24 hours the solvent is evaporated and the residue treated with Et2O (200 ml). The mixture is filtered, the solution was washed with 0.1 M Hcl (2× 100 ml), dried over Na2SO4and evaporated under reduced pressure. The crude product is purified using flash chromatography, obtaining the desired product (13,03 g; 19.2 mmol).

Yield: 32%

K.F.: <0,1%

[α ]20D=-22,47 (1,93, l3)

Elemental analysisNN
% calculated:61,838,456,18
% found:61,70charged 8.52of 5.84

TLC: Stationary phase: a plate of silica gel 60F 254 Merck

Eluent: 3:7=EtOAc/n-g is Xan R f=0,40

Detection: CE(SO4)2·4H2O (0,18%) and (NH4)6Mo7O24·4H2O (3,83%) in 10% H2SO4

Spectra1H-NMR,13C-NMR, IR and MS correspond to the specified structure.

C) Methyl ether (3α , 5β , 7α , 12α )-3-[(chlorocarbonyl)oxy]-12-hydroxypoly-24-OIC acid

WARNING: ALL OPERATIONS SHOULD be PERFORMED IN a WELL-VENTILATED FUME hood

a 20% Solution of phosgene in toluene (100 ml; 202,2 mmol) is added dropwise to a solution of methyl ester (3α , 5β , 7α , 12α )-3,12-dihydroxyfuran-24-OIC acid (14,7 g; 36 mmol) in anhydrous CH2Cl2(350 ml), cooled to 0° C in nitrogen atmosphere. The solution is stirred for 3 hours at room temperature, then evaporated (CAREFULLY)to give the desired product (15.2 g; 32,4 mmol).

Yield: 90%.

Spectra1H-NMR,13C-NMR, IR and MS correspond to the specified structure.

C) Methyl ester [3α (S), 5β , 12α ]-3-[[[[5-[[2-[bis[2-(1,1-dimethylmethoxy)-2-oxoethyl]amino]ethyl][2-(1,1-dimethylmethoxy)-2-oxoethyl]amino]-6-methoxy-6-oxohexyl]amino]carbonyl]oxy]-12-hydroxypoly-24-OIC acid

5% Pd/C (1.3 g) are added to a solution of the compound obtained in stage A) (12.3 g; 18,1 mmol), Meon (120 ml), and the suspension is stirred for 3 hours in hydrogen atmosphere at room temperature. P the following filtration through a Millipore filter® (FT 0.45 µm) solution is evaporated under reduced pressure. The crude product is dissolved in anhydrous CH2Cl2(20 ml) and added dropwise to a solution of compound obtained in stage C) (8.7 g; 18,54 mmol) and N,N-diisopropylethylamine (DIEA) (6,5 ml; 37,07 mmol) in anhydrous CH2Cl2(200 ml) at 0° and in nitrogen atmosphere. After 3 hours the reaction mixture was washed with H2About (2× 100 ml), the organic phase is allocated, dried over Na2SO4and evaporated under reduced pressure. The crude product is purified using flash chromatography, obtaining the desired product (8.6 g; 8,8 mmol).

Yield: 49%.

K.F.:<0,1%

Elemental analysisNN
% calculated:65,079,384,30
% found:65,679,524,24

TLC: Stationary phase: plate silica gel 60 F 254 Merck

Eluent: 3:7=EtOAc/n-hexane Rf=0,30

Detection: CE(SO4)2·4H2O (0,18%) and (NH4)6Mo7O24·4H2O (3,83%) in 10% H2SO4

Spectra1H-NMR,13C-NMR, IR and MS correspond to the specified structure.

D) [3α (S), 5β , 12α ]-3-[[[[5-[[2-[bis(carboxymethyl)amino]ethyl](carboxymethyl)amino]-5-carboxypentyl]amino]carbonyl]oxy]-12-hydroxypoly-24-OIC acid is the

2 M LiOH (133 ml, 266 mmol) are added to a solution of the compound obtained in stage C) (8.5 g; at 10.64 mmol) in 1,4-dioxane (130 ml). The resulting solution was stirred at room temperature for 24 hours, and then neutralized to pH 7 by adding 2 N Hcl (120 ml). The solution is concentrated and viparita under reduced pressure to half volume and slowly acidified to a pH of 1.5 with vigorous stirring, obtaining a white precipitate, which is filtered off, washed with N2About (2× 100 ml) and dried, obtaining the desired product (6,45 g; 8,13 mmol).

Yield: 76%. TPL: 188,9°

K.F.: 1,44%

[α ]20D=+35,47 (2,01, 1 M NaOH)

HPLC analysis: 96,8% (area %)

Stationary phase: Bond Eclipse XDB-C8 3.5 µm; 150× 4.6 mm column filled Rockland Technologies Inc.

Temperature: 40° C;

Spectra1H-NMR,13C-NMR, IR and MS correspond to the specified structure.

(E) a Complex of manganese sodium salt [3α (S), 5β , 12α ]-3-[[[[5-[[2-[bis(carboxymethyl)amino]ethyl](carboxymethyl)amino]-5-carboxypentyl]amino]carbonyl]oxy]-12-hydroxypoly-24-OIC acid (1:3)

The compound obtained in stage D) (5 g; 6.3 mmol), suspended in N2About (50 ml) and dissolved by adding 1 M NaOH (18 ml, 18 mmol). Add 0,559 M aqueous solution of MnCl2(11,27 ml; 6.3 mmol) over 3 hours, maintaining the pH at 6.5 by 1 M NaOH (7.8 ml). Bring the solution pH to 6.8, to whom avlee 1 M NaOH (0.9 ml; 0.9 mmol), filtered (membrane 0.45 µm Millipore®) and absoluut by nanofiltration. The solution is evaporated and dried, obtaining the desired product (of 5.45 g, 6.05 mmol).

Yield: 96%. So pl.: >300°

K.F.: 3,78%

[α ]

20
D
=+2,74 (2, N2O)

CE analysis: 100% (area %)

Capillary: quartz glass 0,72 m × 50 µm

Voltage: 30 kV

Buffer: 0,07 M borate, pH of 9.3, 0.3 mm EDTA

Temperature: 25°

Stop time: 20 min

Detection (UV): 200 nm

Injection: hydrodynamic (50 mbar, 4 sec)

Concentration of sample: 1 mg/ml

Instrument: Hewlett Packard 3D NRSE

Temporary scheme prior training
t (min)action
2rinsing H2O
2rinsing with 0.1 M NaOH
1wash N2About
5flushing the buffer
Elemental analysisHNMnNaCL
% calculated:52,00 of 6.49of 4.666,107,66 
% found:49,547,17of 4.44the 5.657,58<0,1

Spectra IR and MS correspond to the specified structure.

EXAMPLE 16

A complex of gadolinium with a sodium salt of N2bis-[2-[bis(carboxymethyl)amino]ethyl]-N-[(3β , 5β , 7α , 12α )-7,12-dihydroxy-24-oxo-24-[(2-sulfoethyl)amino]Holan-3-yl]-L-glutamine (1:3)

A) Fenilmetilovy ether [3β (S), 5β , 7α , 12α ]-3-[[4-[bis[2-[bis[2-[1,1-dimethylmethoxy)-2-oxoethyl]amino]ethyl]amino]-5-(1,1-dimethylmethoxy)-1,5-dioxaphetyl]amino]-7,12-dihydroxyfuran-24-OIC acid

1-(1,1-dimethylethylene) ether N,N-bis[2-[bis[2-(1,1-dimethylmethoxy)-2-oxoethyl]amino]ethyl]-L-glutamato acid (Anelli, P. L. et al. Bioconjugate Chem. 1999, 10, 137) (37 g; 50 mmol), fenilmetilovy ether [3β , 5β , 7α , 12α ]-3-amino-7,12-dihydroxyfuran-24-OIC acid (Anelli, P. L.; Lattuada, L.; Uggeri, F. Synth. Commun. 1998, 28, 109) (31 g; 55 mmol) and diethylthiophosphate (commercial product) (9.6 g; 55 mmol; 9,2 ml) dissolved in DMF (750 ml). The resulting solution was cooled to 0° C and added dropwise Et3N (7.3 ml). After 1 hour at room temperature the solution is evaporated under reduced pressure, the residue is dissolved in EtOAc (300 ml), washed with 5% aqueous Panso3 (2× 200 ml) and then brine (2× 200 ml). The organic phase is allocated, dried over Na2SO4and then evaporated under reduced pressure. The crude product is purified using flash chromatography, obtaining the desired product (36 g; 29 mmol).

The output 58%

K.F.: 0,98%

Elemental analysisNN
% calculated:65,64of 9.214,57
% found:66,319,204,51

TLC: Stationary phase: a plate of silica gel 60F 254 Merck

Eluent: 2:8=EtOAc/n-hexane Rf=0,3

Detection: Asón/conc. H2SO4/p-anisic aldehyde=100:2:1

Spectra1H-NMR,13C-NMR, IR and MS correspond to the specified structure.

In) Triethylammonium salt [3β (S), 5β , 7α , 12α ]-3-[[4-[bis[2-[bis[2-(1,1-dimethylmethoxy)-2-oxoethyl]amino]ethyl]amino]-5-(1,1-dimethylmethoxy)-1,5-dioxaphetyl]amino]-7,12-dihydroxyfuran-24-OIC acid

5% Pd/C (3.6 g) are added to a solution of the compound obtained in stage A) (36 g, 29.4 mmol)in EtOH (1.5 l) and the suspension is stirred for 3 hours in hydrogen atmosphere at room temperature. After filtration through a Millipore filter® FT 0.45 µm) solution is evaporated under reduced pressure. The crude product is purified using flash HRO is ecografia, getting the right product (22 g; 18 mmol).

An output of 60%. TPL: 58°

K.F.: 1,34%

Elemental analysisNN
% calculated:65,079,865,66
% found:64,3410,075,48

TLC: Stationary phase: a plate of silica gel 60F 254 Merck

Eluent: 1:5:95=Et3N/MeOH/CH2Cl2Rf=0,33

Detection: Asón/conc. H2SO4/p-anisic aldehyde=100:2:1

Spectra1H-NMR,13C-NMR, IR and MS correspond to the specified structure.

C) N2bis[2-[bis(carboxymethyl)amino]ethyl]-N-[(3β , 5β , 7α , 12α )-7,12-dihydroxy-24-oxo-24-[(2-sulfoethyl)amino]Holan-3-yl]-L-glutamine

Et3N (1.2 g; 12 mmol; 1.7 ml) is added dropwise to a solution of compound B) (12.5 g; 11 mmol), 2-aminoethanesulfonic acid (commercial product) (1.5 g, 12 mmol) and diethylthiophosphate (commercial product) (2.1 g; 12 mmol; 2 ml) in DMF (400 ml) at 0° and in nitrogen atmosphere. After 20 min the reaction mixture was left to warm to room temperature and stirred for 3 hours. The solution is evaporated under reduced pressure, the residue is dissolved in dioxane (250 ml) and added dropwise to 0.5 M of water (H2SO4(250 ml, 125 mmol). Obtained from the ect heated at 90° C for 2 hours. Bring the solution pH from 1.4 to 7 by adding 2 N. NaOH, and the solvent is evaporated under reduced pressure. The crude product is purified using flash chromatography. The product is dissolved in N2O (250 ml) and 2 N. Hcl (12.5 ml) and absoluut, elwira gradient of CH3SP/N2About through the column with resin Amberlite® XAD-16.00, obtaining the desired product (1.5 g, 1.6 mmol).

Exit 14%. TPL: >200°

K.F.: by 5.87%

Elemental analysisNNS
% calculated:53,687,447,283,33
% found:50,347,716,642,99

Spectra1H-NMR,13C-NMR, IR and MS correspond to the specified structure.

D) gadolinium Complex with the sodium salt of N2bis[2-[bis(carboxymethyl)amino]ethyl]-N-[(3β , 5β , 7α , 12α )-7,12-dihydroxy-24-oxo-24-[(2-sulfoethyl)amino]Holan-3-yl]-L-glutamine (1:3)

Product C) (880 mg; 0,915 mmol) is dissolved in N2O (50 ml) and 1 N. NaOH is added dropwise to achieve pH values of 6.8. Add Gd2O3(165 mg; 0.46 mmol) and the resulting suspension heated at 50° C for 6 hours. The reaction mixture is filtered through a machine Millipore® filter 0.45 μm) and obtained the filtrate is introduced into a column with a weak cation exchange resin Dowex® CCR 3LB (PA +form, 20 ml). The eluate is evaporated under reduced pressure and dried, obtaining the desired product (1,00 g; 0.75 mmol).

Yield 82%. TPL:> 250°

K.F.: 13,95%

HPLC analysis: 100% (area %)

Stationary phase: Lichrospher 100 RP-8, 5 μm; 250× 4 mm column filled with Merck KGaA

Temperature: 40°

Mobile phase: Isocratic elution pre-mixed mobile phase; 1 g n-octylamine added to 300 ml of acetonitrile, mixed with 700 ml of water. The solution will bufferinput to pH 6 by adding N3RHO4

Flow rate: 1 ml/min

Detection (UV): 200 nm

Elemental analysisNNSGdNa
% calculated:43,785,54of 5.922,7113,30of 5.83
% found:36,826,045,112,18at 10.645,35

Spectra IR and MS correspond to the specified structure.

In a similar way we obtain a gadolinium complex of N2bis[2-[bis(carboxymethyl)amino]ethyl]-N-[(3β , 5β )-24-oxo-24-[(2-sulfoethyl)amino]Holan-3-yl]-L-glutamine.

EXAMPLE 17

A complex of gadolinium with a sodium salt of [3α (S), 5β ]-3-[2-[[5-[bis[2-[bis(carboxyl who yl)amino]ethyl]amino]-5-carboxypentyl]amino]-2-oksidoksi]Holan-24-OIC acid (1:3)

A) Methyl ester [3α , 5β ]-3-(carboxymethoxy)Holan-24-OIC acid

Triplet of benzylglycine (Williams, M.A.; Rapoport, H.J. Org. Chem. 1994, 59, 3616) (16,8 g; 56,3 mmol) is added over a period of time longer than 15 minutes to a stirred solution of methyl ester [3α , 5β ]-3-hydroxyfuran-24-OIC acid (Dayal, V. et al. Steroids, 1981, 37, 239) (20 g; a 51.2 mmol) and N,N-diisopropylethylamine (10 ml; 57,4 mmol) in anhydrous CH3SP (400 ml) at -20° C. After 4 hours at -20° the mixture is heated to room temperature and stirred for further 4 hours. The solvent is evaporated and the residue partitioned between EtOAc (300 ml) and saturated NaHCO3(300 ml). The organic phase is dried over Na2SO4and evaporated. The residue is dissolved in EtOH (200 ml), then add 5% Pd/C (3 g) and the mixture is stirred at room temperature for 5 hours in an atmosphere of hydrogen. After filtration through a Millipore filter® FH (0.45 µm) solution is evaporated and the residue purified using flash chromatography, obtaining the desired product (14.2 g; and 31.7 mmol).

Yield: 62%

K.F.:<0,1

Elemental analysisN
% calculated:72,289,89
% found:71,979,81

Spectra1H-NMR,13C-NMR, IR and MS, which correspond to this structure.

In) [3α (S), 5β ]-3-[2-[[5-[bis[2-[bis(carboxymethyl)amino]ethyl]amino]-5-carboxypentyl]amino]-2-oksidoksi]Holan-24-OIC acid

(1,1-Dimethylethylene) ether N2N2bis[2-[bis[2-[1,1-dimethylmethoxy)-2-oxoethyl]amino]ethyl]-L-lysine (Anelli, P. L. et al. Bioconjugate Chem. 1999, 10, 137) (18,56 g; 25 mmol), the compound (A) (11.2 g; 25 mmol) and diethylthiophosphate (commercial product) (4.9 g; 28 mmol) was dissolved in DMF (300 ml). The resulting solution was cooled to 0° and added dropwise Et3N (4 ml). After 6 hours at room temperature the solution is evaporated under reduced pressure, the residue is dissolved in EtOAc (250 ml), washed with 5% aqueous Panso3(2× 100 ml) and then brine (2× 100 ml). The organic phase is allocated, dried over Na2SO4and then evaporated under reduced pressure. The residue is dissolved in dioxane (300 ml) and added dropwise to 0.5 M water H2SO4(300 ml; 150 mmol). The resulting mixture is heated at 90° C for 2 hours. Bring the solution pH to 7 by adding 2 N. NaOH, and the solvent is evaporated under reduced pressure. The crude product is purified using flash chromatography. The product is dissolved in N2O (300 ml) and 2 N. Hcl (30 ml) and absoluut, elwira through the column with resin Amberlite® XAD-16.00 gradient of CH3SN/N2Oh, obtaining the desired product (9,03 g; of 10.25 mmol).

Exit 41%

K.F.: 2,78%

Elemental analysisNN
% calculated:59,988,246,36
% found:58,118,286,14

Spectra1H-NMR,13C-NMR, IR and MS correspond to the specified structure.

C) gadolinium Complex with the sodium salt of [3α (S), 5β ]-3-[2-[[5-[bis[2-[bis(carboxymethyl)amino]ethyl]amino]-5-carboxypentyl]amino]-2-oksidoksi]Holan-24-OIC acid (1:3)

Product) (5 g; 5,67 mmol) is dissolved in N2O (200 ml), adding 1 N. NaOH to achieve a pH of 6.8. Add Gd2O3(1,03 g; 2,84 mmol) and the resulting suspension heated at 50° C for 8 hours. The reaction mixture was filtered through a Millipore apparatus® (0.45 µm filter) and the filtrate is evaporated under reduced pressure, dried, obtaining the desired product (5,74 g; a total of 5.21 mmol).

Yield 92%. TPL: >250°

K.F.: 5,81%

Elemental analysisNNGdNa
% calculated:47,996,045,09of 14.286,26
% found:45,096,154,7713,39of 5.81

Spectra IR and Sootvetstvuut this structure.

Similarly, based on the compound (A) of example 15, receive gadolinium complex [3α (S), 5β ]-3-[2-[[5-[[2-[bis(carboxymethyl)amino]ethyl](carboxymethyl)amino]-5-carboxypentyl]amino]-2-oksidoksi]Holan-24-OIC acid

Similarly, based on the compound (A) of example 15 and methyl ether (3β , 5β , 7α , 12α )-3-[(3-carboxy-1-oxopropyl)amino]-7,12-dihydroxyfuran-24-OIC acid (obtained according to the method, opened in WO-A-95/32741: example 12), receive gadolinium complex [3β (S), 5β , 7α , 12α ]-3-[[4-[[5-[[2-[bis(carboxymethyl)amino]ethyl](carboxymethyl)amino]-5-carboxypentyl]amino]-1,4-DIOXOLANYL]amino]-7,12-dihydroxyfuran-24-OIC acid.

EXAMPLE 18

Measurements of the relaxation rate (Δ 1/T1)

The effectiveness of the compounds of the present invention as agents for blood depot assessed by plotting the relaxation rate 1/T1depending on the time elapsed after injection. The relaxation rate of the protons 1/T1blood samples obtained at predefined points in time, measured at a temperature of 39° With the instrument Bruker Minispec PC120, using the three-parameter sequence “inversion recovery”.

The compound obtained in example 1, a complex of gadolinium with 1-deoxy-1-(methylamino)-D-glaciology salt [3β (S), 5β ]-3-[[4-[bis[2-[bis(carboxymethylamino]ethyl]amino]-4-carboxy-1-oxobutyl]amino]Holan-24-OIC acid (1:3), administered to rabbits in a dose of 0.1 mmol/kg In the diagram (Fig.1) shows a plot of the relaxation rate.

The compound obtained in example 9 of the patent application WO 95/32741, a complex of gadolinium with 1-deoxy-1-(methylamino)-D-glaciology salt [3β , 5β , 7α , 12α ]-3-[[N-[N-[2-[[2-[bis(carboxymethyl)amino]ethyl](carboxymethyl)amino]ethyl]-N-(carboxymethyl)glycyl]glycyl]amino]-7,12-dihydroxyfuran-24-OIC acid (1:2), enter the rabbit at a dose of 0.1 mmol/kg In the diagram (Figure 2) shows a plot of the relaxation rate.

The compound obtained in example 15 the patent application WO 95/32741, a complex of gadolinium with 1-deoxy-1-(methylamino)-D-glaciology salt [3β (S), 5β , 7α , 12α ]-3-[[4-[bis[2-[bis(carboxymethyl)amino]ethyl]amino]-4-carboxy-1-oxobutyl] amino]-7,12-dihydroxyfuran-24-OIC acid (1:3), enter the rabbit at a dose of 0.1 mmol/kg In the diagram (Figure 3) shows a plot of the relaxation rate.

The compound obtained in example 4, a complex of gadolinium with 1-deoxy-1-(methylamino)-D-glaciology salt [3β (S), 5β , 12α ]-3-[[4-[bis[2-[bis(carboxymethyl)amino]ethyl]amino]-4-carboxy-1-oxobutyl]amino]-12-hydroxypoly-24-OIC acid (1:3), enter the monkey at a dose of 0.05 mmol/kg In the diagram (Figure 4) shows a plot of the relaxation rate.

EXAMPLE 19

0.3 M pharmaceutical composition of gadolinium complex of 1-deoxy-1-(methylamino)-D-glaciology salt [3β (S), 5; , 12α ]-3-[[4-[bis[2-[bis(carboxymethyl)amino]ethyl]amino]-4-carboxy-1-oxobutyl]amino]-12-hydroxypoly-24-OIC acid (1:3)

31,775 kg of gadolinium complex of 1-deoxy-1-(methylamino)-D-glaciology salt [3β (S), 5β , 12α ]-3-[[4-[bis[2-[bis(carboxymethyl)amino]ethyl]amino]-4-carboxy-1-oxobutyl]amino]-12-hydroxypoly-24-OIC acid (1:3) (obtained by the method of example 4) and 100 g of tramadolhydrochloride dissolved in 100 l of sterile water at room temperature in the pharmaceutical reactor made of stainless steel. After dissolution, the pH of the solution was adjusted to 7.4 by adding 1 M trometamol. The solution is filtered under sterile conditions through filters with a diameter of 0.22 mm and distribute in 20 ml ampoules which close halogenosilanes stoppers, sealed with aluminum rings and sterilized with steam at Fo=18. HPLC shows 0,294 M title.

EXAMPLE 20

Pre-filled plastic syringes containing the composition of example 19.

Portions of 20 ml of the solution prepared in example 19, is placed in CZ plastic syringe, the tip of which is closed by the tip. The piston is inserted under vacuum, and the pre-filled syringe is sterilized in an autoclave to a value of Fo=18.

Diagnostic agent containing chelated complexes of ions of two or three-valent paramagnetic metal selected from the group consisting of Fe(2+)(3+), C(2+), Cr(3+), Gd(3+), Eu(3+), Dy(3+), Yb(3+)or MP(2+)with compounds of the formula (I)and their physiologically compatible salts of organic bases selected from primary, secondary, tertiary amines or basic amino acids, or inorganic bases, cations are sodium, potassium, magnesium, calcium or mixtures thereof:

X-L-Y (I)

where X represents the residue polyaminocarboxylic ligand or its derivative selected from the group consisting of ethylendiaminetetraacetic acid (EDTA), diethylenetriaminopentaacetic acid (DTPA), 1,4,7,10-tetraazacyclododecane-1,4,7,10-tetraoxane acid (DOTA), 1,4,7,10-tetraaza-cyclododecane-1,4,7-trioxanes acid (DO3A), [10-(2-hydroxypropyl)-1,4,7,10-tetraazacyclododecane-1,4,7-trioxanes acid (HPDO3A), 4-carboxy-5,8,11-Tris(carboxy-methyl)-1-phenyl-2-oxa-5,8,11-trustregion-13-OIC acid (WORTH);

Y is derived bile acids selected from the group consisting of residues holeva, chenodeoxycholic, deoxycholic, ursodeoxycholic, lithocholic acid:

as such, as well as funktsionirovanii on the provisions containing a hydroxyl group as a reactive group, regardless of the stereochemistry of the final products, and specified production the second also includes the conjugate acid group in position 24 with taurine and glycine;

L is a chain associated with any position of X, not necessarily including one of the carboxyl groups, which thus turns into an amide group, and C-3, C-7, C-12 Y-positions, and has the following formula (II):

where m is an integer from 1 to 10, where values greater than 1 And can have different values,

And represented by the following formula (III):

n and q can be 0 or 1 but cannot both be zero,

R can take values in the interval from 0 to 10,

Z represents an oxygen atom or an-NR-group, in which R represents a hydrogen atom or a (C1-C5)alkyl group, unsubstituted or substituted by-COOH group,

to obtain diagnostic compositions for NMR imaging the circulatory system of humans and animals.

2. The diagnostic agent according to claim 1, where the complexes are formed with ions of gadolinium or manganese.

3. The diagnostic agent according to claim 1, where in formula (I) connecting chain L has formula (IIIa) and (IIIb):

4. The diagnostic agent according to claims 1-3, where Z represents an oxygen atom, and L is thus formed due to the hydroxyl groups present in 3, 7, 12 positions, regardless stereohype the final products.

5. The diagnostic agent according to claim 3, where the residue X is selected from the group consisting of EDTA, DTPA, DOTA, DO3A, WORTH; L is selected from the group consisting of (IIIa) and (IIIb) and Y is selected from the group consisting of residues holeva, deoxycholic, chenodeoxycholic, lithocholic acid as they are or in which one or more of the hydroxyl groups were converted into ketogroup associated with L amino group in position 3, and an acid group at position 24 is present as such or as a derivative of taurine or glycine; complexes of these compounds formed with ions of gadolinium or manganese, and the cations are suitable for the neutralization of organic bases selected from the group consisting of ethanolamine, diethanolamine, research, glucamine, N-methylglucamine, N,N-dimethylglycine, or cations of inorganic bases selected from the group consisting of sodium, potassium, magnesium, calcium or mixtures thereof.

6. The diagnostic agent according to claim 5, formed by a compound of General formula (IV)

where in the formula (I) residue X is DTPA, substituted in the Central chain, and where R1represents a hydrogen atom or-COOH group, Y is selected from the group consisting of residues holeva, deoxycholic, chenodeoxycholic, lithocholic acid,

L is represented by structural formula (III).

7. Diagnostic Agay is t 6, formed by a compound of General formula (IVa)

where Y takes the values specified above for the compounds of General formula (IV)

L represents the structural formula (IIIa) and (IIIb).

8. The diagnostic agent according to claim 7 where the compound of General formula (IVa) is selected from the group consisting of

[3(β(S), 5β]-3-[[4-[bis[2-[bis(carboxymethyl)amino]ethyl]amino]-4-carboxy-1-oxobutyl](carboxymethyl)amino]Holan-24-OIC acid;

[3β(S), 5β]-3-[[4-[bis[2-[bis(carboxymethyl)amino]ethyl]amino]-4-carboxy-1-oxobutyl]amino]Holan-24-OIC acid;

[3β(S), 5β]-3-[[4-[bis[2-[bis(carboxymethyl)amino]ethyl]amino]-4-carboxy-1-oxobutyl]amino]-12-Oklahoman-24-OIC acid;

[3β(S), 5β, 7α]-3-[[4-[bis[2-[bis(carboxymethyl)amino]ethyl]amino]-4-carboxy-1-oxobutyl]amino]-7-hydroxypoly-24-OIC acid;

N2bis[2-[bis(carboxymethyl)amino]ethyl]-N-[(3β, 5β)-24-oxo-24-[(2-sulfoethyl)amino]Holan-3-yl]-L-glutamine;

N2bis[2-[bis(carboxymethyl)amino]ethyl]-N-[(3β, 5β, 7α, 12α)-7,12-dihydroxy-24-oxo-24-[(2-sulfoethyl)amino]Holan-3-yl]-L-glutamine;

[3β(S), 5β, 7β]-3-[[4-[bis[2-[bis(carboxymethyl)amino]ethyl]amino]-4-carboxy-1-oxobutyl]amino]-7-hydroxypoly-24-OIC acid;

[3β(R), 5β, 12α]-3-[[4-[bis[2-[bis(carboxymethyl)amino]e is Il]amino]-4-carboxy-1-oxobutyl]amino]-12-hydroxypoly-24-OIC acid;

[3β(RS), 5β, 12α]-3-[[4-[bis[2-[bis(carboxymethyl)amino]ethyl]amino]-4-carboxy-1-oxobutyl]amino]-12-hydroxypoly-24-OIC acid;

[3β(RS), 5β, 7α, 12α]-3-[[4-[bis[2-[bis(carboxymethyl)amino]ethyl]amino]-4-carboxy-1-oxobutyl]amino]-7,12-dihydroxy-Holan-24-OIC acid;

[3α(S), 5β, 7α, 12α]-3-[[[[5-[bis[2-[bis(carboxymethyl)amino]ethyl]amino]-5-carboxypentyl]amino]carbonyl]hydroxy-7,12-dihydroxyfuran-24-OIC acid;

[3α(S), 5β]-3-[2-[[5-[bis[2-[bis(carboxymethyl)amino]ethyl]amino]-5-carboxypentyl]amino]-2-oksidoksi]Holan-24-OIC acid;

[3β(S), 5β, 7α, 12α]-3-[[4-[bis[2-[bis(carboxymethyl)amino]ethyl]amino]-4-carboxy-1-oxobutyl]amino]-7,12-dihydroxyfuran-24-OIC acid;

[3β(S), 5β, 7α, 12α]-3-[[4-[[5-[bis[2-[bis(carboxy-methyl)amino]ethyl]amino]-5-carboxypentyl]amino]-1,4-DIOXOLANYL]amino]-7,12-dihydroxyfuran-24-OIC acid.

9. The diagnostic agent according to claim 6, formed by a compound of General formula (IVb)

where Y takes the values specified above for the compounds of General formula (IV)

L represents the structural formula (IIIa).

10. The diagnostic agent according to claim 9, where the compound of General formula (IVb) is selected from the group consisting of

(3β, 5β, 7α, 12α)-3-[[[bis[2-[bis(carboxymethyl)amino]the Tyl]amino]acetyl]amino]-7,12-dihydroxyfuran-24-OIC acid;

(3β, 5β)-3-[[[[[bis[2-[bis(carboxymethyl)amino]ethyl]amino]acetyl]amino]acetyl]amino]Holan-24-OIC acid;

(3β, 5β, 7α, 12α)-3-[[[[[bis[2-[bis(carboxymethyl)amino]ethyl]amino]acetyl]amino]acetyl]amino]-7,12-dihydroxyfuran-24-OIC acid;

(3β, 5β, 7α, 12α)-3-[[6-[[[bis[2-[bis(carboxymethyl)amino]ethyl]amino]acetyl]amino]-1-oxohexyl]amino]-7,12-dihydroxyfuran-24-OIC acid.

11. The diagnostic agent according to claim 5, formed by a compound of General formula (V)

where in the General formula (I) residue X is DTPA,

Y takes the values specified above for the compounds of General formula (IV)

L represents the structural formula (IIIa).

12. The diagnostic agent according to claim 11 where the compound of General formula (V) selected from the group consisting of

(3β, 5β, 7α, 12α)-3-[[N-[N-[2-[[2-[bis(carboxymethyl)amino]ethyl](carboxymethyl)amino]ethyl]-N-(carboxymethyl)glycyl]glycyl]amino]-7,12-dihydroxyfuran-24-OIC acid;

18-[[(3β, 5β, 7α, 12α)-23-carboxy-7,12-hydroxy-24-norolean-3-yl]amino]-3,6,9-Tris(carboxymethyl)-11,18-dioxo-3,6,9,12-tetraazabicyclo acid.

13. The diagnostic agent according to claim 5, formed by a compound of General formula (VI)

where in the General formula (I) residue X PR is dstable DO3A,

Y takes the values specified above for the compounds of General formula (IV)

L is selected from formulas (IIIa) and (IIIb).

14. The diagnostic agent according to claim 11 where the compound is 10-[3-[[(3α, 5β, 7α, 12α)-23-carboxy-7,12-dihydroxy-24-norolean-3-yl]oxy]-2-hydroxy-propyl]-1,4,7,10-tetraazacyclododecane-1,4,7-trioxanes acid.

15. The diagnostic agent according to claim 5, formed by a compound of General formula (VII)

where in the General formula (I) residue X is EDTA,

Y takes the values specified above for the compounds of General formula (IV)

L represents a group of formula (III).

16. The diagnostic agent according to item 15, where the compound of General formula (VII) is selected from the group consisting of

[3α(S), 5β, 12α]-3-[[[[5-[[2-[bis(carboxymethyl)amino]ethyl](carboxymethyl)amino]-5-carboxypentyl]amino]carbonyl]oxy]-12-hydroxypoly-24-OIC acid;

[3β(S), 5β, 7α, 12α]-3-[[4-[[5-[[2-[bis (carboxymethyl)amino]ethyl](carboxymethyl)amino]-5-carboxypentyl]amino]-1,4-DIOXOLANYL]amino]-7,12-dihydroxyfuran-24-OIC acid;

[3β(S), 5β]-3-[2-[[5-[[2-[bis(carboxymethyl)amino]ethyl](carboxymethyl)amino]-5-carboxypentyl]amino]-2-oksidoksi]Holan-24-OIC acid;

[3β(S), 5β, 12α]-3-[[4-[[2-[[bis(carboxymethyl)amino]ethyl](carboxymethyl)amino]-4-carboxy-1-OK is Outil]amino]-12-hydroxypoly-24-OIC acid;

[3β(S), 5β]-3-[[4-[[2-[[bis(carboxymethyl)amino]ethyl](carboxymethyl)amino]-4-carboxy-1-oxobutyl]amino]-12-Oklahoman-24-OIC acid.

17. The diagnostic agent according to claim 1 to 16, where the chelate complex salts formed with sodium and N-methylglucamine.

18. The intermediate compound of General formula (IVa)

where Y is selected from the remnants of holeva, deoxycholic, chenodeoxycholic, lithocholic acid as such or such, in which one or more hydroxyl groups were converted into ketogroup associated with L amino group in position 3, and an acid group at position 24 is present as such or in the form of its touringwagon or glycine derivative; and complexes of these compounds formed with ions of gadolinium or manganese and base cations suitable for neutralization selected from the group consisting of ethanolamine, diethanolamine, research, glucamine, N-methylglucamine, N,N-dimethylglycine, or from group consisting of inorganic bases, cations are sodium, potassium, magnesium and calcium,

L has the structural formula (IIIa) and (IIIb)

where m is an integer from 1 to 10,

with the exception of [3β(S), 5β, 7α, 12α]-3-[[4-[bis[2-[bis(carboxy-methyl)amino]ethyl]amino]-4-carboxy-1-about libutil]amino]-7,12-dihydroxyfuran-24-OIC acid and [3β (S), 5β, 7α, 12α]-3-[[4-[[5-[bis[2-[bis(carboxymethyl)amino]ethyl]amino]-5-carboxypentyl]amino]-1,4-DIOXOLANYL]amino]-7,12-dihydroxyfuran-24-OIC acid.

19. The intermediate connection p selected from the group consisting of

[3β(S), 5β]-3-[[4-[bis[2-[bis(carboxymethyl)amino]ethyl]amino]-4-carboxy-1-oxobutyl](carboxymethyl)amino]Holan-24-OIC acid;

[3β(S), 5β]-3-[[4-[bis[2-[bis(carboxymethyl)amino]ethyl]amino]-4-carboxy-1-oxobutyl]amino]Holan-24-OIC acid;

[3β(S), 5β]-3-[[4-[bis[2-[bis(carboxymethyl)amino]ethyl]amino]-4-carboxy-1-oxobutyl]amino]-12-Oklahoman-24-OIC acid;

[3βand(3), 5β, 7α]-3-[[4-[bis[2-[bis(carboxymethyl)amino]ethyl]amino]-4-carboxy-1-oxobutyl]amino]-7-hydroxypoly-24-OIC acid;

N2bis[2-[bis(carboxymethyl)amino]ethyl]-N-[(3β, 5β)-24-oxo-24-[(2-sulfoethyl)amino]Holan-3-yl]-L-glutamine;

N2bis[2-[bis(carboxymethyl)amino]ethyl]-N-[(3β, 5β, 7α, 12α)-7,12-dihydroxy-24-oxo-24-[(2-sulfoethyl)amino]Holan-3-yl]-L-glutamine;

[3β(S), 5β, 7β]-3-[[4-[bis[2-[bis(carboxymethyl)amino]ethyl]amino]-4-carboxy-1-oxobutyl]amino]-7-hydroxypoly-24-OIC acid;

[3β(R), 5β, 12α]-3-[[4-[bis[2-[bis(carboxymethyl)amino]ethyl]amino]-4-carboxy-1-oxobutyl]amino]-12-hydroxypoly-24-OIC acid;

[3β(RS), 5β, 12± ]-3-[[4-[bis[2-[bis(carboxymethyl)amino]ethyl]amino]-4-carboxy-1-oxobutyl]amino]-12-hydroxypoly-24-OIC acid;

[3β(RS), 5β, 7α, 12α]-3-[[4-[bis[2-[bis(carboxymethyl)amino]ethyl]amino]-4-carboxy-1-oxobutyl]amino]-7,12-dihydroxy-Holan-24-OIC acid;

[3α(S), 5β, 7α, 12α]-3-[[[[5-[bis[2-[bis(carboxymethyl)amino]ethyl]amino]-5-carboxypentyl]amino]carbonyl]oxy]-7,12-dihydroxyfuran-24-OIC acid;

[3α(S), 5β]-3-[2-[[5-[bis[2-[bis(carboxymethyl)amino]ethyl]amino]-5-carboxypentyl]amino]-2-oksidoksi]Holan-24-OIC acid.

20. The intermediate compound of General formula (IVb)

where Y takes the values specified in p,

L takes values corresponding to formula (IIIa)as defined in p,

except (3β, 5β, 7α, 12α)-3-[[[[[bis[2-[bis(carboxymethyl)amino]ethyl]amino]acetyl]amino]acetyl]amino]-7,12-dihydroxyfuran-24-OIC acid and (3β, 5β, 7α, 12α)-3-[[6-[[[bis[2-[bis(carboxymethyl)amino]ethyl]amino]acetyl]amino]-1-oxohexyl]amino]-7,12-dihydroxyfuran-24-OIC acid.

21. The intermediate connection according to claim 20, selected from the group consisting of

(3β, 5β, 7α, 12α)-3-[[[bis[2-[bis(carboxymethyl)amino]ethyl]amino]acetyl]amino]-7,12-dihydroxyfuran-24-OIC acid;

(3β, 5β)]-3-[[[[[bis[2-[bis(carb is kemetyl)amino]ethyl]amino]acetyl]amino]acetyl]amino]Holan-24-OIC acid.

22. The intermediate compound of General formula (VII)

where Y takes the values specified in p,

L denotes the formula (III) as defined in claim 1.

23. The intermediate connection p.22 selected from the group consisting of

[3α(S), 5β, 12α]-3-[[[[5-[[2-[bis(carboxymethyl)amino]ethyl](carboxymethyl)amino]-5-carboxypentyl]amino]carbonyl]oxy]-12-hydroxypoly-24-OIC acid;

[3β(S), 5β, 7α, 12α]-3-[[4-[[5-[[2-[bis (carboxymethyl)amino]ethyl](carboxymethyl)amino]-5-carboxypentyl]amino]-1,4-DIOXOLANYL]amino]-7,12-dihydroxyfuran-24-OIC acid;

[3β(S), 5β]-3-[2-[[5-[[2-[bis(carboxymethyl)amino]ethyl](carboxymethyl)amino]-5-carboxypentyl]amino]-2-oksidoksi]Holan-24-OIC acid;

[3β(S), 5β, 12α]-3-[[4-[[2-[[bis (carboxymethyl)amino]ethyl](carboxymethyl)amino]-4-carboxy-1-oxobutyl]amino]-12-hydroxypoly-24-OIC acid;

[3β(S), 5β]-3-[[4-[[2-[[bis(carboxymethyl)amino]ethyl](carboxymethyl)amino]-4-carboxy-1-oxobutyl]amino]-12-Oklahoman-24-OIC acid.

24. Contratulations diagnostic pharmaceutical composition comprising at least one of the diagnostic agents according to claims 1 to 17 or its salt according to claims 1 and 4.

25. The pharmaceutical composition according to paragraph 24, which includes, in addition, not more than 5 mol % chelate complex is a salt of calcium or magnesium and/or physiologically acceptable buffer, selected from trometamol and glycylglycine.

26. The method of obtaining compounds of formula (IV) according to the following scheme:

where R4is aminosidine group,

R5represents an unbranched or branched C1-C10alkyl or aryl,

R2and R3independently represent a hydrogen atom;

this method is characterized by the fact that in the original pyrrolidone introduce a protective group, R4and R5capable chipped off in different conditions, then the action of an amine of the formula R-NH2where R is Y, conduct the reaction transaminirovania received a secure connection, allowing you to save the stereochemistry at the chiral center adjacent to the nitrogen atom of the original pyrrolidone, after which the product transaminirovania enter into reaction with the compound of the formula

with the removal of the protective group R4and the formation of the corresponding N,N-disubstituted amino, and further, if necessary, removing the protective groups present in the fragment of R, to obtain the final product.

27. The method according to p obtain compounds

[3β(S), 5β, 7α, 12α]-3-[[4-[bis[2-[bis(carboxymethyl)amino]ethyl]amino]-4-carboxy-1-oxobutyl]amino]-12-hydro is sholan-24-OIC acid and [3β (S), 5β, 7α, 12α]-3-[[4-[bis[2-[bis(carboxy-methyl)amino]ethyl]amino]-4-carboxy-1-oxobutyl]amino]-7,12-dihydroxyfuran-24-OIC acid.



 

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