Classifications

• • A—HUMAN NECESSITIES
  • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
  • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
  • A61K49/00—Preparations for testing in vivo
  • A61K49/06—Nuclear magnetic resonance [NMR] contrast preparations; Magnetic resonance imaging [MRI] contrast preparations
  • A61K49/08—Nuclear magnetic resonance [NMR] contrast preparations; Magnetic resonance imaging [MRI] contrast preparations characterised by the carrier
  • A61K49/10—Organic compounds
• • A—HUMAN NECESSITIES
  • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
  • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
  • A61K49/00—Preparations for testing in vivo
  • A61K49/04—X-ray contrast preparations
  • A61K49/0433—X-ray contrast preparations containing an organic halogenated X-ray contrast-enhancing agent
  • A61K49/0438—Organic X-ray contrast-enhancing agent comprising an iodinated group or an iodine atom, e.g. iopamidol
• • A—HUMAN NECESSITIES
  • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
  • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
  • A61K49/00—Preparations for testing in vivo
  • A61K49/06—Nuclear magnetic resonance [NMR] contrast preparations; Magnetic resonance imaging [MRI] contrast preparations
  • A61K49/08—Nuclear magnetic resonance [NMR] contrast preparations; Magnetic resonance imaging [MRI] contrast preparations characterised by the carrier
  • A61K49/085—Nuclear magnetic resonance [NMR] contrast preparations; Magnetic resonance imaging [MRI] contrast preparations characterised by the carrier conjugated systems
• • A—HUMAN NECESSITIES
  • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
  • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
  • A61K49/00—Preparations for testing in vivo
  • A61K49/06—Nuclear magnetic resonance [NMR] contrast preparations; Magnetic resonance imaging [MRI] contrast preparations
  • A61K49/08—Nuclear magnetic resonance [NMR] contrast preparations; Magnetic resonance imaging [MRI] contrast preparations characterised by the carrier
  • A61K49/10—Organic compounds
  • A61K49/101—Organic compounds the carrier being a complex-forming compound able to form MRI-active complexes with paramagnetic metals
  • A61K49/103—Organic compounds the carrier being a complex-forming compound able to form MRI-active complexes with paramagnetic metals the complex-forming compound being acyclic, e.g. DTPA
• • C—CHEMISTRY; METALLURGY
  • C07—ORGANIC CHEMISTRY
  • C07F—ACYCLIC, CARBOCYCLIC OR HETEROCYCLIC COMPOUNDS CONTAINING ELEMENTS OTHER THAN CARBON, HYDROGEN, HALOGEN, OXYGEN, NITROGEN, SULFUR, SELENIUM OR TELLURIUM
  • C07F9/00—Compounds containing elements of Groups 5 or 15 of the Periodic Table
  • C07F9/02—Phosphorus compounds
  • C07F9/28—Phosphorus compounds with one or more P—C bonds
  • C07F9/38—Phosphonic acids [RP(=O)(OH)2]; Thiophosphonic acids ; [RP(=X1)(X2H)2(X1, X2 are each independently O, S or Se)]
  • C07F9/3804—Phosphonic acids [RP(=O)(OH)2]; Thiophosphonic acids ; [RP(=X1)(X2H)2(X1, X2 are each independently O, S or Se)] not used, see subgroups
  • C07F9/3808—Acyclic saturated acids which can have further substituents on alkyl
• • C—CHEMISTRY; METALLURGY
  • C07—ORGANIC CHEMISTRY
  • C07F—ACYCLIC, CARBOCYCLIC OR HETEROCYCLIC COMPOUNDS CONTAINING ELEMENTS OTHER THAN CARBON, HYDROGEN, HALOGEN, OXYGEN, NITROGEN, SULFUR, SELENIUM OR TELLURIUM
  • C07F9/00—Compounds containing elements of Groups 5 or 15 of the Periodic Table
  • C07F9/02—Phosphorus compounds
  • C07F9/547—Heterocyclic compounds, e.g. containing phosphorus as a ring hetero atom
  • C07F9/553—Heterocyclic compounds, e.g. containing phosphorus as a ring hetero atom having one nitrogen atom as the only ring hetero atom
  • C07F9/572—Five-membered rings
  • C07F9/5728—Five-membered rings condensed with carbocyclic rings or carbocyclic ring systems
• • C—CHEMISTRY; METALLURGY
  • C07—ORGANIC CHEMISTRY
  • C07J—STEROIDS
  • C07J51/00—Normal steroids with unmodified cyclopenta(a)hydrophenanthrene skeleton not provided for in groups C07J1/00 - C07J43/00
• • Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
  • Y10—TECHNICAL SUBJECTS COVERED BY FORMER USPC
  • Y10S—TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
  • Y10S977/00—Nanotechnology
  • Y10S977/902—Specified use of nanostructure
  • Y10S977/904—Specified use of nanostructure for medical, immunological, body treatment, or diagnosis
  • Y10S977/927—Diagnostic contrast agent
  • Y10S977/93—MRI contrast agent

Definitions

• This invention refers to new compounds which can chelate paramagnetic bi- and trivalent metal ions, their chelates with said metal ions and their use as contrast agents in magnetic resonance imaging (M.R.I.) .
• an improvement in the radiographic image which means a better contrast enhancement between healthy and diseased tissues, is seen as an aid to the diagnosis which can be obtained through previous administration of suitable exogenous substances.
• contrast agents for M.R.I.
• a number of chelated complexes of linear and cyclic polyaminopolycarboxylic ligands with paramagnetic metals are known to be useful as M.R.I, contrast agents.
• Said compounds generally derive from the two basic polyaminopolycarboxylic structures, namely diethylenetriaminopentaacetic acid (DTP A) and l,4,7,10-tetraazacyclododecane-l,4,7,10-tetraacetic acid (DOTA).
• the compounds of the present invention are novel polyamino derivatives comprising at least one phosphonic residue as one of the binding site in the chelating agent structure.
• Relaxivity is an intrinsic property of paramagnetic complexes which characterizes their ability to increase the nuclear magnetic relaxation rate of vicinal protons.
• Gd(III) chelates with q ⁇ 1, wherein q is the number of coordinated water molecules, a remarkable contribution to the increase in relaxation observed for water protons of the solvent derives from the exchange between the molecule(s) of bound water and the molecules of the remaining solvent (S. Aime et al., Chem. Soc. Rev., 1998, 27, 19).
• This contribution ( ⁇ ) is related to the relaxation time (T w ) and to the residence time ( ⁇ ) of the protons of the water molecule(s) which are coordinated in the inner coordination sphere according to the following equation:
• T w receives contributions from the reorientation of the paramagnetic species, ⁇ R ,_through the residence time of the coordinated water protons, ⁇ M , and the electronic relaxation time of the metal ion, ⁇ g .
• r" p is the highest when T w > ⁇ M (fast exchange conditions) and T w is as short as possible.
• the residence time of the water molecule(s) coordinated to a Gd(III) complex plays a particularly important role, in that it directly contributes to the nucleus-electron dipolar interaction and controls the transfer efficiency of the paramagnetic effect to the water molecules of the solvent.
• ⁇ M values generally comprised between 200 and 2500 ns, where such values are significantly higher than the 30 ns optimum one. Optimization and harmonization of the above parameters are still remarkably important objects for everyone dealing with the development of novel M.R.I, contrast agents.
• the present invention relates to polyamino derivatives comprising as the binding site in the structure of the chelating agent at least one phosphonic residue, capable of causing an increase in the proton exchange rate and therefore advantageously low ⁇ values.
• the object of the present invention is acyclic polyamino derivative chelating agents of formula (I), both in the racemic and the optically active forms, wherein
• R is a hydrogen atom, or -(CH 2 ) m -0-R 2 , (C ⁇ -C 5 )-alkyl-(C 6 -C ⁇ o)- aryl or (C 1 -C 5 )-alkyl-heteroaryl whose aryl or heteroaryl moiety comprises 1 or 2 fused rings optionally substituted with one or more halogen atoms, OH groups, alky ⁇ C ⁇ Cs) groups and/or an OR3 group, wherein R 2 is (Ci[-C5)-alkyl-(C6-C ⁇ o)- ar yl > optionally substituted with one or more halogen atoms, OH and (C ⁇ -C5)-alkyl groups;
• R3 is (Cg-Cio) aryl optionally substituted with one or more halogen atoms, OH and/or (C ⁇ -C5)-alkyl groups; m ranges from 1 to 5;
• R ⁇ can have the same meanings as R with the proviso that when Y is PO(OH) 2 , Ri is selected from H, (CH 2 ) m NH2 5 (CH2) m COOH or an amido derivative thereof.
• a further object of the invention are the chelates of said compounds of formula (I) with the bi- and trivalent ions of metal elements having atomic number ranging between 20 and 31, 39, 42, 43, 44, 49, or between 57 and 83, as well as the salts thereof with physiologically compatible organic bases selected from primary, secondary, tertiary amines or basic amino acids, or with inorganic bases whose cations selected from sodium, potassium, magnesium, calcium or mixtures thereof.
• a further object of the present invention is the use of the compounds of formula (I), their complexes with paramagnetic metals and the physiologically compatible salts thereof for the preparation of pharmaceutical formulations for the imaging of organs and/or tissues of the human or animal body, by use of M.R.L.
• Examples of (C ⁇ -C5)-alkyl-(C6-C ⁇ o)-aryl groups comprise benzyl, phenethyl, naphthylmethyl wherein the aryl moiety is optionally substituted with one or more halogen atoms or OR3 groups wherein R3 is as defined above.
• Examples of (C 1 -C 5 )-alkyl-heteroaryl groups comprise pyridylmethyl or indolylm ethyl.
• Examples of (Cg-Cjo) aryl groups comprise phenyl or naphthyl optionally substituted with one or more halogen atoms, OH and/or (C1-C5)- alkyl groups.
• Examples of (C1-C5) alkyl groups preferably comprise methyl, ethyl, isopropyl.
• R and Ri have the above defined meanings.
• compounds of formula (II) particularly preferred are those in which Ri; is an hydrogen atom and R can assume all previously defined meanings. Also preferred are the compounds of formula (III)
• Rl has all the values defined above, as well as the compounds of general formula (IV),
• R has the values defined above. Particularly preferred are the following compounds:
• Preferred chelates are those in which the bi- or trivalent metal ion is selected from Gd (3+) , Dy (3+) , Fe (3+) , Fe (2+) and Mn (2+) . Particularly preferred are
• Gd (3+) chelates Preferred cations of inorganic bases optionally suitable for salifying the chelated complexes of the invention particularly comprise the ions of alkali or alkaline-earth metals such as potassium, sodium, calcium, magnesium, and mixtures thereof.
• Preferred cations of organic bases suitable for this purpose comprise, inter alia, those obtained by protonation of primary, secondary and tertiary amines such as ethanolamine, diethanolamine, morpholine, glucamine, N- methylglucamine, N,N-dimethylgluc amine.
• Preferred cations of amino acids comprise, for example, those of lysine, arginine or ornithine.
• the chelated complexes of the invention are characterized by ⁇ ⁇ 100 ns values, preferably values between 10 and 100 ns, most preferably between 20 and 50 ns.
• ⁇ ⁇ 100 ns values preferably values between 10 and 100 ns, most preferably between 20 and 50 ns.
• R has the values defined above for compounds (I).
• the synthetic process of Scheme 1 comprises t the following steps: a) Esterif ⁇ cation of a suitable amino acid. Said esterif ⁇ cation can be advantageously carried out by reacting the amino acid with an alkyl acetate and an acid such as HCIO4.
• the amino acid previously N-protected by reaction with CBZC1
• reaction is carried out in an organic solvent preferably selected among acetonitrile, THF, EtOAc and in the presence of a pH 8 buffer solution; c) Bromoalkylation of the intermediate 2 by reacting it with trifluoromethanesulfonic acid 2-bromoethyl ester (intermediate 3) previously prepared from bromoethanol, trifluoromethanesulfonic anhydride and 2,6- lutidine.
• organic solvent preferably selected among acetonitrile, THF, EtOAc and in the presence of a pH 8 buffer solution
• intermediate 3 previously prepared from bromoethanol, trifluoromethanesulfonic anhydride and 2,6- lutidine.
• the bromoalkylation is carried out in an organic solvent suitably selected among, for example, toluene, acetonitrile, dichloroethane, and in the presence of an amine selected among ethylenediamine, diisopropylethylamine, triethylamine, to give the intermediate 4.
• organic solvent suitably selected among, for example, toluene, acetonitrile, dichloroethane, and in the presence of an amine selected among ethylenediamine, diisopropylethylamine, triethylamine, to give the intermediate 4.
• compound 4 can be alternatively prepared starting from the corresponding N- (2 -hydroxy ethyl) derivative, obtained as described in WO 98/05625, (incorporated herein by reference in its entirety), by reacting it with a brominating agent such as NBS, in the presence of triphenylphosphine; d) Preparation of aminomethylphosphonic acid diethyl ester (intermediate 5) by direct condensation of tribenzylhexahydrotriazine with a suitable dialkyl phosphite and subsequent debenzylation by catalytic hydrogenation of the condensation product; e) Bis alkylation of intermediate 5 by reaction with intermediate 4 and isolation of hexaester 6.
• a brominating agent such as NBS
• the bis alkylation reaction is preferably carried out in an organic solvent such as acetonitrile, ethyl acetate, and in the presence of a pH 8 buffer solution; f) Deprotection of the acidic functions of intermediate 6 and isolation of the acid chelating agent 7. Said deprotection can be obtained by reacting the hexaester with iodotrimethylsilane in an organic solvent, such as CH3CN.
• the synthetic process of SCHEME Ibis essentially comprises the following steps: a) Preparation of 2,2'-(Iminodi-2,l-ethanediyl)bis-lH-isoindole-l,3(2H)- dione) (intermediate 1) by reacting phthalic anhydride with diethylenetriamine in acetic acid; b) N-alkylation of the bis-phthalimido derivative 1 by reacting it with 3-benzyloxycarbonylpropionaldehyde (intermediate 2) in a suitable organic medium, and then with tris (tert-butyl) phosphite to give intermediate 3; c) Removal of phthalic groups to give corresponding diamine (intermediate 4) by reaction, for example, with hydrazine; d) N-alkylation of the diamine 4 by reaction with a suitable halo acetate, such as, for example, tert-butyl bromoacetate, to give intermediate 5.
• This reaction is carried out in an organic solvent preferably selected from acetonitrile, ethyl acetate, and in the presence of a suitable tertiary amine such as, for example, diisopropylethylamine; e) Debenzylation by catalytic hydrogenation of the intermediate 5 and isolation of the hexaester monocarboxylic acid 6. In a preferred process this hydrogenation is carried out in an organic solvent such as, for example, THF and catalysed by 10% Pd-C; f) reaction of the hexaester monocarboxylic acid 6 with a suitable amino compound (compound 7) and isolation of the corresponding amide (derivative 8).
• an organic solvent preferably selected from acetonitrile, ethyl acetate, and in the presence of a suitable tertiary amine such as, for example, diisopropylethylamine; e) Debenzylation by catalytic hydrogenation of the intermediate 5 and isolation of the he
• said reaction is performed in presence of HATU (0-(7-Azabenzotriazol- 1 -yl)-N,N,N' ,N'-tetramethyluronium hexafluoro-phosphate); g) Deprotection of the acidic functions of the hexaester and recovery of the acid chelating agent (compound 10).
• the deprotection of the acidic functions is performed on the hexaester derivative prepared at step e) (intermediate 6) to give, for example, the acid chelating agent of EXAMPLE 7, disclosed later on in the experimental section of the invention.
• the deprotection includes a first debenzylation step, by catalytic hydrogenation, of the benzylester contained in the amido derivative 8 and a second step, including the deprotection of the residual acidic functions of the hexaester 9, to give the chelating agent 10.
• the hydrogenation is preferentially carried out in an organic solvent such as, for example, THF and catalysed by 10% Pd-C.
• the subsequent deprotection can be performed, for example, by reacting the hexaester 9 with trifluoroacetic acid.
• the synthetic process of Scheme 2 comprises the following steps: a) Preparation of aminomethylphosphonic acid bis tert-butyl ester (bis N-alkyl) derivative (intermediate 3) by reacting bis tert-butyl phosphite, suitably activated (intermediate 1), with aminal (intermediate 2).
• the phosphonic acid tert-butyl ester is advantageously activated for example with Me3SiCl in a reaction carried out in organic solvent and in the presence of an amine, such as triethylamine.
• the resulting trimethylsilyl derivative is reacted with intermediate 2 obtained from 2-benzylaminoethanol and aqueous formaldehyde.
• This reaction is activated by the presence of a catalytic amount of a lanthanide triflate. Particularly preferred is ytterbium triflate.
• the resulting trimethylsilyl derivative is not isolated but it is directly transformed into the corresponding hydroxy derivative by treatment with a suitable aqueous acid, such as aqueous acetic acid.
• a suitable aqueous acid such as aqueous acetic acid.
• step b) N-alkylation of the resulting compound from step b), (intermediate 4), by reacting it with a suitable haloacetate, such as tert-butyl bromoacetate.
• a suitable haloacetate such as tert-butyl bromoacetate.
• This reaction is carried out in an organic solvent preferably selected from acetonitrile, ethyl acetate, and in the presence of a buffer solution pH 8.
• This reaction is carried out in an organic solvent selected from THF, acetonitrile, ethyl acetate, under nitrogen atmosphere and in the presence of an amine selected from triethylamine, diisopropylethylamine, at temperatures ranging from 20 to -5°C.
• an organic solvent selected from THF, acetonitrile, ethyl acetate, under nitrogen atmosphere and in the presence of an amine selected from triethylamine, diisopropylethylamine, at temperatures ranging from 20 to -5°C.
• Said reaction is advantageously carried out in an organic solvent preferably selected from acetonitrile, THF, ethyl acetate and in the presence of a buffer solution pH 8.
• the compounds of this invention have a wide range of applications, since they can be used for intravasal, (for instance i.v., intraarterial, intracoronaric, intraventricular administration and so on), intrathecal, intraperitoneal, intralymphatic and intracavital administrations. Furthermore, the compounds are suitable for the oral or enteral administration, and therefore, specifically for the imaging of the gastrointestinal tract.
• aqueous solutions or suspensions whose pH can range from 6.0 to 8.5.
• aqueous solutions or suspensions can be administered in concentrations ranging between 0.002 and 1.0 M.
• These formulations can be lyophilized and supplied as they are to be reconstituted before use.
• these agents can be formulated as a solution or suspension optionally containing suitable excipients in order, for example, to control viscosity.
• suitable excipients for example, to control viscosity.
• they can be formulated according to preparation methods routinely used in the pharmaceutical technique or as coated formulations to gain additional protection against the stomach acidic pH, thus preventing the chelated metal ion from release, which takes place particularly at the pH values typical of gastric juices.
• excipients such as sweeteners and/or flavouring agents, can also be added, according to known techniques of pharmaceutical formulations.
• solutions or suspensions of the compounds of this invention can also be formulated as aerosols to be used in aerosol-broncho graphy and instillation.
• the compounds of the present invention can optionally be chemically conjugated to suitable macromolecules, targeting vectors or inglobated into suitable carriers.
• they can also be encapsulated in liposomes or they can be constituents of their chemical structure and used as uni- or multilamellar vesicles.
• TLC Carrier: silica gel plate 60F 254 Merck Eluent: CHCI3/CH3OH 95:5.
• TLC Carrier: silica gel plate 60F 254 Merck
• the intermediate prepared at point C) (147.2 g; 573 mol) is added under nitrogen to a solution of N-[2-(l,l-dimethylethoxy)-2-oxoethyl]-L- phenylalanine 1,1 -dimethyl ethyl ester (65.93 g; 197 mol) and 2,6-lutidine (72 mL; 0.62 mol) in 600 mL of dry toluene at -15°C. After 16 h at room temperature, 200 mL of EtOAc, 200 mL of H2O and 50 mL of ethylenediamine are added to the mixture.
• TLC Carrier: silica gel plate 60F 254 Merck
• the mixture is then cooled to room temperature, taken up with ethyl ether (150 mL) and acidified with 6N HCl (20 mL).
• the organic phase is extracted with IN HCl (10 mL), the aqueous phases are basified with 5N KOH, then extracted with Et 2 0 (300 + 150 mL), washed with brine (100 mL) and finally dried over Na2S ⁇ 4- After evaporation of the solvent under vacuum, the product is recovered as a colourless oil (25.05 g; 97.37 mol) which is stored at a temperature of -18°C.
• TLC Carrier: silica gel plate 60F 254 Merck
• TLC Carrier: silica gel plate 60F 254 Merck
• TLC Carrier: silica gel plate 60F 254 Merck
• intermediate 4 is prepared as follows: to a solution of N-[2-(l,l-dimethylethoxy)-2-oxoethyl]-N-(2- hydroxyethyl)-0-(phenylmethyl)-L-serine 1,1-dimethylethyl ester (prepared as described in WO 98/05625) (61.7 g; 150.7 mol) and triethylamine (31 L; 0.22 mol) in dry THF (600 mL) are slowly added, under nitrogen atmosphere, methanesulfonyl chloride (12.5 mL; 160 mol) and lithium bromide (111 g; 1.25 mol) at -15/-10°C.
• the solvent is evaporated under vacuum to obtain the product as a colourless oil (44.84 g; 47.19 mol).
• a jacketed reactor is loaded with a solution of N-[2-(l,l- dimethylethoxy)-2-oxoethyl]-L-tryptophan 1,1-dimethylethyl ester (5 g; 13.35 mol) prepared at point B, in CH3CN (25 mL).
• ethylene oxide 13 mL; 0.26 mol
• ytterbium triflate 0.83 g; 1.34 mol
• TLC Carrier: silica gel plate 60F 254 Merck
• Residual methanol is evaporated by azeotropic distillation with cyclohexane to obtain the desired intermediate as a colourless oil (39.45 g) which, although containing traces of solvents, can be used (as it is) in the subsequent reaction without further purification.
• TLC Carrier: silica gel plate 60F 254 Merck
• polyester E (24.9 g; 19.6 mol) in 1,4-dioxane (100 mL) is added 4N HCl (180 mL). The solution is heated to 70°C for 3 hours then at 90°C for 1 hour, following the progress of the reaction by HPLC. The mixture is cooled to room temperature, concentrated to 200 mL and slowly
• HPLC assay 99.5% (in % area)
• the product is prepared by reacting the aminophosphonate (15.09 g; 90.28 mol) prepared as described in IF and the bromide (70.09 g; 199.0 mol) prepared as described in J. Org. Chem. 1993, 58, 1151. The reaction is carried out as reported in 1G and the product is recovered as a colourless oil (35.59 g;
• the hexaester prepared at point A (29.23 g; 41.18 mol) is deprotected with iodomethylsilane, under the same conditions as reported in 1H.
• the resulting crude is loaded onto a Relite 3 AS/fb column which is eluted with water until elimination of the residual iodide and then onto a column of
• step B To a solution of the chelating agent obtained as in step B (6.0 g; 13.97 mol) in H2O (60 mL) is added 2N NaOH (15 mL) and Gd2 ⁇ 3 (2.53 g; 6.99 mol). The suspension is heated at 70°C for one hour, then filtered through
• the resulting hexaester is deprotected under the same conditions as reported in 4F.
• the isolated acidic chelating agent is complexed according to the procedure reported in 3G.
• An analytical sample is crystallized from ethanol.
• a solution of the 3-benzyloxycarbonylpropionaldehyde (4.0 g; 0.0208 • mol) in acetonitrile (25 mL) is added to a slurry of bis(phthalimido) derivative prepared at point A) (6.8 g; 0.0187 mol) in acetonitrile (75 mL) over a period of 30 min.
• the temperature of the reaction mixture is maintained at 80-90°C during the addition.
• the reaction mixture is stirred at 80°C for an additional 30 min.
• the colour of the reaction mixture turns yellow during the addition of the aldehyde.
• Tris(tert-butyl) phosphite (5.2 g; 0.0208 mol) in acetonitrile (15 mL) is added dropwise and the reaction mixture is stirred at room temperature for 48 h. The reaction mixture becomes a clear yellow solution after the addition of tris(tert- butyl)phosphite ( ⁇ 3 h). Acetonitrile is then removed and the residue is treated with EtOAc (50 mL). The solid formed is filtered and the EtOAc solution is directly applied to a column of silica gel (packed in 50:50 hexane- EtOAc).
• the acid chelating ligand TFA salt obtained at step F) (0.25 g) is dissolved in a mixture of acetonitrile water 1:1 (5 mL) and Gd (OAc) 3 (0.25 g; 0.006 mol) is then added to the reaction mixture.
• the initial pH ofthe reaction mixture is found to be 1.29.
• the pH of the solution is then adjusted to 5.0 by the addition of IN NaOH and the mixture is stirred at room temperature for 24 h.
• the Gd complex is then purified by DEAE Sephadex chromatography using triethylamine bicarbonate buffer. The fractions eluted at 800-mM buffer are collected and freeze dried to give the Gd complex as a triethylamine salt (0.2 g).
• HPLC assay 100% (in % area) Elemental Analysis: Calcd. for C 16 H 25 N 3 0 13 PGd ⁇ 2.6 C 6 H 15 N • 3.H 2 0 C H Gd N H 2 0
• Trifluoroacetic acid (4 mL) is added to the hexa-(tert-butyl) ester (0.9 g; 0.75 mol) prepared at point B) and the mixture is stirred at room temperature for 12 h. Diethylether (30.0 mL) is then added to the reaction mixture and the precipitate is filtered and dried under vacuum to give a white solid (0.8 g) (Yield: 97%).
• HPLC analysis of the TFA salt indicates that it is fairly pure and useful for Gd chelation without further purification. An analytical sample is obtained from TFA salt (100 mg) by preparative HPLC. Fractions containing the pure product are collected and freeze dried to give a white fluffy solid (50 mg). Purification yield: 50% Elemental analysis (calcd.for C 4 ⁇ H 55 F 3 N 5 0 17 P * 2H 2 0)
• TFA salt obtained as disclosed at point C (0.3 g; 0.275 mol) is added to a mixture of acetonitrile (7.0 mL) and water (2 mL).
• a solution of Gd(OAc) 3 (0.132 g; 0.325 mol) in water (2.0 mL) is added dropwise.
• the initial pH ofthe solution is found to be 1.29.
• the solution becomes turbid and the pH of the solution is adjusted to 5.0 by adding a solution of meglumine in water.
• the turbid reaction mixture is stirred at room temperature for 48 h.
• the pH of the reaction mixture is then raised to 9.0 by the addition of meglumine solution and then purified by preparative HPLC using acetonitrile and water. Fractions containing the pure product were collected and freeze dried to give a fluffy solid (220 mg).. Yield: 78.5 %
• EXAMPLE 7 (1.2 g; 1.4 mol) in CH 2 C1 2 is added HATU (0.6 g; 1.5 mol) and the mixture is stirred at 0.°C for 10 min.
• Diisopropylethylamine (1 mL) is then added and the mixture is stirred at 0°C for further 10 min.
• 3 ⁇ -Aminocholic acid methyl ester (compound disclosed in WO 9532741) ( 0.526 g; 1.25 mol) is then added to the mixture and stirred at 0°C for 2 h and at room temperature for 48 h.
• CH 2 C1 2 is removed on a rotary evaporator and the residue is extracted with EtOAc.
• Trifluoroacetic acid (3 mL) is added to the hexa (tert-butyl) ester prepared at step B) (0.662 g; 0.5 mol) and the mixture is stirred at room temperature for 30 min and kept at 4°C for 18 h. Trifluoroacetic acid is then removed to give the desired deprotected intermediate (0.52 g) as an oil which is dried under vacuum.
• To a solution of said compound in ethanol water (1:1,5 mL) is added sodium hydroxide (20%, 5 mL) and the mixture is stirred at room temperature for 24 h. The completion of the reaction is followed by HPLC. The solvents are removed and the residue is neutralized with IN HCl to give the completely deprotected polyacid as hydrochloride, which is filtered and dried under vacuum to give the desired chelating compound (0.35 g).

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• 1999
  • 1999-12-21 IT IT1999MI002656A patent/IT1315263B1/it active
• 2000
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  • 2000-12-20 AU AU30148/01A patent/AU3014801A/en not_active Abandoned
  • 2000-12-20 DE DE60018390T patent/DE60018390T2/de not_active Expired - Lifetime
  • 2000-12-20 US US09/913,711 patent/US6509324B1/en not_active Expired - Fee Related
  • 2000-12-20 JP JP2001547116A patent/JP2003518131A/ja active Pending

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