Patent Application: US-201013389837-A

Abstract:
the invention relates to polysultone derivatives used as precursors to radiolabelled macromolecules usable for medicine and in nuclear imaging . the aim of the invention is to provide novel prosthetic compounds or groupings , the synthesis of which is straightforward , easy and automatable , enabling access to economical and effective radiolabelled macromolecules . the aim is achieved by the invention , which involves compounds of formula 10 or 11 . said double - sultone derivatives are produced by opening the sultone rings using a nucleophile radical r i which may be a radionucleide r * for one of the sultone rings and an active radical peptide r a for the other sultone ring . the invention also relates to the method for producing the abovementioned compounds , as well as to the drugs or diagnosis products that the latter are capable of forming .

Description:
in the general formulae of the novel compounds 1 , 2 , 3 , reference is made to the following definitions : “ alkyl ” corresponds for example to a linear , branched or cyclic saturated monovalent c1 - c30 alkyl group , preferably c1 - c20 , and , even more preferentially c1 - c10 , optionally substituted , comprising or not comprising heteroatoms . examples of alkyl groups are in particular methyl , ethyl , isopropyl , n - propyl , tert - butyl , isobutyl , n - butyl , n - pentyl , isoamyl and 1 , 1 - dimethylpropyl . “ aryl ” corresponds for example to one or more monocyclic or polycyclic and preferably monocyclic or bicyclic condensed or uncondensed aromatic monovalent groups , having 6 to 18 carbon atoms . it must be understood that , within the framework of the invention , by polycyclic aromatic radical is meant a radical having two or more aromatic rings , condensed ( orthocondensed or ortho - and pericondensed ) with each other , i . e . having , in pairs , at least two carbon atoms in common . said aromatic hydrocarbon group (“ aryl ”) is optionally substituted for example by one or more c 1 - c 3 alkyls , one or more halogenated hydrocarbon radicals ( e . g . cf 3 ), one or more alkoxy ( e . g . ch 3 o ) or one or more hydrocarbon radicals comprising one or more ketone units ( e . g . ch 3 co —). by way of examples of aryls , there can be mentioned the phenyl , naphthyl , anthryl and phenanthryl radicals . “ arylalkyl ” corresponds for example to an alkyl group as defined above , substituted by one or more aryl groups on its hydrocarbon chain , the aryl group being as defined above . examples of this are benzyl and triphenylmethyl . “ alkylaryl ” corresponds for example to monovalent alkyl , substituted or linked to one or more monovalent aromatic groups , optionally substituted . by “ acyl ” is meant an r 0 — co — group where r 0 represents alkyl as defined above ; or an ar — co — group where ar represents an aryl group as defined above , or arylalkyl in which aryl and alkyl are as defined above and in which the aryl part is optionally substituted e . g . by alkyl . by “ cycloalkyl ” is meant a mono - or polycyclic , preferably mono - or bicyclic , saturated hydrocarbon radical preferably having from 3 to 10 carbon atoms , even better from 3 to 8 . by polycyclic saturated hydrocarbon radical is meant a radical having two or more cyclic rings attached to each other by σ bonds and / or condensed in pairs . examples of polycyclic cycloalkyl groups are adamantane and norbornane . examples of monocyclic cycloalkyl groups are cyclopropyl , cyclobutyl , cyclopentyl , cyclohexyl , cycloheptyl and cyclooctyl . by “ alkene ” or “ alkenyl ” is meant e . g . a substituted or unsubstituted , linear or branched , unsaturated hydrocarbon chain , having at least one olefinic double bond , and more preferably a single double bond . preferably , the alkenyl group has 2 to 8 carbon atoms , even better 2 to 6 . this hydrocarbon chain optionally comprises at least one heteroatom such as o , n , s . preferred examples of alkenyl groups are the allyl and homoallyl groups . by “ alkyne ” or “ alkynyl ” is meant e . g . according to the invention , a substituted or unsubstituted , linear or branched , unsaturated hydrocarbon chain , having at least one acetylenic triple bond , and more preferably a single triple bond . preferably , the alkynyl group has 2 to 8 carbon atoms , even better 2 to 6 carbon atoms . by way of example , there can be mentioned the acetylenyl group , as well as the propargyl group . this hydrocarbon chain optionally comprises at least one heteroatom such as o , n , s . the spacer can for example be an aromatic structure ( functionalized or not ), an alkyne structure or any other structure . in the simplest cases , the r 1 - r 6 groups are protons ( in particular in the r 6 and r 3 nucleophilic attack positions ) but they can also be any other group . the precursors 1 according to the invention can be disultones with the following formulae : the structure 1a corresponds to the case where the spacer is an aromatic ring linked to the two sultones by two ketone functions in the para position , where the r 1 - r 6 groups are all hydrogens and where n = m = 1 . the structure 1b corresponds to the case where the spacer is an alkyne linked to the two sultones by two ketone functions , where the r 1 - r 6 groups are all hydrogens and where n = m = 1 . the structure 1c corresponds to the case where the spacer is an aromatic ring linked to the two sultones by two alcohol functions , where the r 1 - r 6 groups are all hydrogens and where n = m = 1 . the structure 1d corresponds to the case where the spacer is an aromatic ring linked to the two sultones by two alcohol functions in the para position , where the r 1 - r 6 groups are all hydrogens and where n = 2 and m = 1 . the structure 1e corresponds to the case where the spacer is a tetrafluoroethane unit directly linking the two sultones , where the r 1 - r 6 groups are all fluorines and where n = m = 0 . thus , the method for synthesizing the compound according to the invention essentially consists of reacting sultones — advantageously butane sultones , propane sultones and / or ethane sultones — and for example a polyester , preferably according to a mechanism of lithiation of each alpha - sulphur of the sultone followed by a reaction with the electrophilic polyester which is added to the reaction medium for this purpose . for example , two equivalents of the corresponding sultone and one equivalent of a diester corresponding to the spacer via a reaction of lithiation of the alpha - sulphur of the sultone followed by the reaction with the diester as electrophile . this synthesis is based on the chemistry of the sultones , cyclic compounds which are sulphur - containing lactone analogues with 4 , 5 or 6 members ( fig4 ). the sultones are well known for their reactivity with nucleophiles 11 , 12 . this type of reactivity was moreover already the subject of a patent filed in 2004 13 . the reaction forms a carbon - nucleophile bond by nucleophilic attack on the alpha - carbon of the oxygen and results in the formation of a sulphonate function . the reactivity of the sultones with the nucleophiles such as the cyanides phosphines 14 , 15 , amines 16 , 17 , 18 , alcoholates 16 , 19 thiolates 20 , iodine , chlorine and bromine 19 , 21 , 22 is well known . the synthesis diagram of the compound 1a is represented below in fig5 . the lithiation of the alpha - sulphur of two equivalents of propane sultone at − 78 ° c . in thf over 30 minutes followed by the addition of 1 equivalent of the methyl ( or paranitrophenyl ) diester leads to the corresponding disultone 1a . the examples which follow illustrate this synthesis . the synthesis diagram of the compound 1b is represented below in fig6 . the lithiation of the alpha - sulphur of two equivalents of propane sultone at − 78 ° c . in thf over 30 minutes followed by the addition of 1 equivalent of commercial butynedioic acid methyl diester . the examples which follow illustrate this synthesis . these polysultones , and , in particular , the double sultones 1a and 1b ) can for example be used : either as a “ linker ”, a binding entity , between two nucleophiles or structures bearing a nucleophile by opening of the sultone rings , or according to a preferred but non - limitative embodiment of the invention : for labelling macromolecules ( bearing the nucleophile opening the second sultone ) with a nucleophilic radionuclide ( opening the first sultone ) such as for example fluorine - 18 , bromine - 76 , iodine - 123 , iodine - 131 or any other nucleophilic radionuclide as well as all combinations of nucleophilic radionuclides . the polysultone precursors 1 defined above give access to novel compounds each substituted by at least one nucleophilic radical by opening at least one of the sultone rings , preferably by at least one ( preferably one ) nucleophilic radical r i on each of the opened sultone rings . advantageously , the nucleophilic radical ( s ) r i is ( are ) one or more radionuclide ( s ) r *, preferably selected from the group comprising fluorine - 18 , bromine - 76 , iodine - 123 , iodine - 131 or any other nucleophilic radionuclide . preferably , the nucleophilic radical ( s ) r i is ( are ) one or more active radical ( s ) r a selected from the group comprising the peptides , the proteins , the oligonucleotides , the polynucleotides or any other macromolecule . even more preferentially , by way of example starting from 1a and 1b these compounds correspond to the following formulae 6a or 6b : the more particularly preferred compounds according to the invention are compounds having at least one substitution by a radioactive group and at least one substitution by an active group of macromolecule type ( for example in therapy or in diagnosis ). each of these compounds is characterized in that it is substituted by one or more radionuclide ( s ), preferably selected from the group comprising fluorine - 18 , bromine - 76 , iodine - 123 , iodine - 131 or any other nucleophilic radionuclide , by opening one of the sultone rings and in that it is substituted by a nucleophilic radical selected from the group comprising the peptides , proteins , oligonucleotides , polynucleotides or any other macromolecule by opening the other sultone ring optionally via the attack of a free amine . more precisely , by way of example starting from 1a and 1b these preferred compounds correspond e . g . to the following formulae 7a or 7b respectively : according to the invention , the method for obtaining a novel compound originating from a poly -( e . g . double ) sultone precursor is characterized in that it comprises the following stages : a . utilization or synthesis of a polysultone , preferably a double sultone , by the method according to claim 10 ; b . opening of one of the sultones with a first nucleophile leading to the formation of a sulphonate ; c . separation of the polysultone ( preferably the double sultone ) precursor and of the formed sulphonate by difference in polarity ; d . collection of the formed sulphonate ; e . coupling of the formed sulphonate with at least one second nucleophile by opening a ( preferably the ) second sultone ; f . collection of the polysulphonate ( preferably disulphonate ) obtained in stage e . by way of example , the labelling of macromolecules with fluorine - 18 is represented in fig7 below . the product 1a is radiofluorinated under standard radiofluorination conditions and the resultant ( very polar ) fluorosulphonate 8 is easily separated from the ( apolar ) disultone precursor 1a via a c18 cartridge due to their differences in polarity . the experimental part of the synthesis of the non - radioactive reference [ 19 f ]- 8 and of the radiosynthesis of the product [ 18 f ]- 8 is described in the examples which follow . the obtained fluorosulphonate 8 is then coupled in water ( solubility of the fluorosulphonate in aqueous medium ) with a peptide by opening the second sultone with a free amine borne by the peptide leading to the radiofluorinated peptide 9 . by way of example , the use of the boc - lysine - nh 2 10 makes it possible to obtain the radiofluorinated boc lysine 11 : fig7 . the procedure for the synthesis of [ 19 f ]- 11 and the radiosynthesis of [ 18 f ]- 11 are described in the examples which follow . this labelling strategy can also be extended to the labelling of the boc - lysine - nh 2 10 with iodine . the iodized analogue 12 is obtained by coupling the boc - lysine 10 with the iodized sultone 13 , itself obtained by radioiodination of the disultone 1a . ( see fig9 below ). a description of the procedure for the synthesis of 13 by non - radioactive chemistry is appended in the examples which follow . in the same way , the use of the rgd amino peptide 14 as a labelling precursor makes it possible to obtain the radiofluorinated peptide 15 : fig1 . this labelling strategy can also be extended to the labelling of the rgd peptide 14 with iodine . the iodized analogue 16 is obtained by coupling the peptide precursor 14 with the iodized sultone 13 , itself obtained by radioiodination of the disultone 1a . a description of the procedure for the synthesis of 13 by non - radioactive chemistry is appended in the experimental part . this labelling strategy will therefore make it possible to have the same single method for labelling , for example , a peptide with iodine - 123 ( for use in spect ) or with fluorine - 18 ( for use in pet ). system a : analytical hplc ( hypersil gold c 18 column , 5 μm , 4 . 6 × 100 mm ) with ch 3 cn and 0 . 1 % trifluoroacetic acid in water as eluents ( 0 . 1 %, v / v , ph 2 . 0 ) [ 100 % tfa ( 5 min ), then linear gradient from 0 % to 80 % ( 40 min ) with ch 3 cn ] at a flow rate of 1 ml / min . the uv detections are carried out at 265 and 254 nm . system b : semi - preparative hplc ( hypersil gold c 18 column , 5 μm , 21 . 2 × 250 mm ) with ch 3 cn and 0 . 1 % trifluoroacetic acid in water as eluents ( 0 . 1 %, v / v , ph 2 . 0 ) [ 100 % tfa ( 10 min ), then linear gradient from 0 % to 5 % with ch 3 cn ( 2 min ), then linear gradient from 5 % to 70 % in ch 3 cn ( 65 min )] at a flow rate of 1 ml / min . the uv detections are carried out at 265 and 254 nm . system c : analytical hplc ( hypersil gold c 18 column , 5 μm , 4 . 6 × 100 mm ) with ch 3 cn and 25 mm of triethylammonium acetate in water as eluents ( teaa , 25 mm , ph 7 . 0 ) [ 100 % teaa ( 10 min ), then linear gradient from 0 % to 80 % ( 40 min ) with ch 3 cn ] at a flow rate of 1 ml / min . the uv detections are carried out in the max - plot mode ( each peak is detected at its absorption maximum ). system d : semi - preparative hplc ( hypersil gold c 18 column , 5 μm , 10 × 250 mm ) with ch 3 cn and 50 mm of aqueous trimethylammonium bicarbonate as eluents ( teab , 50 mm , ph 7 . 5 ) [ 100 % teab ( 10 min ), then linear gradient from 0 % to 5 % ( 2 min ) with ch 3 cn , then linear gradient from 5 % to 18 % with ch 3 cn ( 13 min ), then linear gradient from 18 % to 25 % ( 14 min ) with ch 3 cn , then linear gradient from 25 % to 31 % ( 6 min ) with ch 3 cn , then linear gradient from 31 % to 61 % ( 15 min ) with ch 3 cn ] at a flow rate of 1 ml / min . the uv detection is carried out at 254 and 220 nm . a 1 . 3 m solution of nbuli in hexane ( 2 . 82 ml , 3 . 67 mmol , 1 . 99 eq ) is added dropwise to a solution of 1 , 3 - propane sultone ( 447 mg , 3 . 66 mmol , 2 eq ) in thf ( 5 ml ), under an argon atmosphere , at − 78 ° c . after 1 h at − 78 ° c ., a solution of dimethyl terephthalate ( 358 mg , 1 . 84 mmol , 1 eq ) in 10 ml of thf is added dropwise to the vigorously stirred previous mixture . the mixture is then stirred at − 78 ° c . for 2 h 30 then , still at − 78 ° c ., the reaction is quenched with 1 ml of glacial acetic acid in 3 ml of thf ( ph ˜ 6 ). the mixture is then slowly warmed to at then diluted in 20 ml of saturated nacl in water and 50 ml of ethyl acetate . a solid is observed between the organic phase and the aqueous phase and is filtered off . the aqueous phase is washed with 30 ml of ethyl acetate . the organic phases are combined , dried with mgso 4 , filtered and concentrated under vacuum . the crude product obtained is then purified by flash chromatography on a silica column with an acetone - cyclohexane mixture ( 2 : 3 , v / v ) as mobile phase . the bis - sultone , obtained is isolated in the form of a beige solid ( 270 mg , yield 40 %). r f 0 . 42 ( acetone - cyclohexane , 2 : 3 , v / v ); 1 h nmr ( 300 mhz , acetone - d 6 ,): δ 8 . 20 ( s , 4h ), 5 . 63 - 5 . 57 ( m , 2h ), 4 . 59 - 4 . 49 ( m , 4h ), 3 . 17 - 3 . 08 ( m , 2h ), 2 . 80 - 2 . 71 ( m , 2h ). 13 c nmr ( 75 mhz , dmso - d6 ): δ 195 . 4 , 140 . 8 , 128 . 4 , 63 . 3 , 58 . 8 , 32 . 2 uv ( recorded during the hplc analysis ): λ max 265 nm ; ms ( esi , negative mode ): m / z 373 . 27 [ m + h ] − c 14 h 14 o 8 s 2 374 . 01 200 μl of a solution of potassium fluoride in mq water ( 62 . 2 mg / ml , 12 . 44 mg , 0 . 214 mmol , 1 eq ), kryptofix ( 95 mg , 0 . 252 mmol , 1 . 18 eq ) and 1 ml of acetonitrile are introduced into a flask . the bis - sultone 1a ( 80 mg , 0 . 214 mmol , 1 eq ) in solution in 5 . 5 ml of acetonitrile is then added . the mixture is stirred at at and progress is monitored using hplc ( system a ). the crude product of the reaction is purified by semi - preparative hplc . after identification by mass spectrometry , the fraction containing the product is lyophilized . the product is obtained in the form of a beige solid ( 27 . 4 mg , 32 . 5 %). 1 h nmr ( 300 mhz , meod - d 3 ,): δ 8 . 16 - 8 . 09 ( m , 4h ), 5 . 56 - 5 . 50 ( ddd , j = 2 . 46 , 6 . 6 , 8 . 67 hz , 1h ), 5 . 07 - 5 . 02 ( ddd , j = 0 . 96 , 3 . 96 , 9 . 42 hz , 1h ), 4 . 61 - 4 . 46 ( m , 2h ), 4 . 45 - 4 . 37 ( m , 1h ), 4 . 29 - 4 . 21 ( m , 1h ), 3 . 15 - 3 . 04 ( m , 1h ), 2 . 71 - 2 . 58 ( m , 1h ), 2 . 55 - 2 . 33 ( m , 2h ). hplc ( system a ): t r = 18 . 8 ( dial )− 19 ( dia2 ) min ( purity 85 %); uv ( recorded during the hplc analysis ): λ max 265 nm ; ms ( esi , negative mode ): m / z 393 . 13 [ m − h ] − c 14 fh 15 o 8 s 2 394 . 02 boc - lys - nh 2 ( 21 . 5 mg , 0 . 088 mmol , 1 . 1 eq ) in dmf ( 0 . 8 ml ) is added to a solution of the fluorinated sultone 8 ( 31 . 5 mg , 0 . 08 mmol , 1 eq ) in dmf ( 1 . 8 ml ), at ambient temperature . the reaction is monitored by hplc ( system a ). once the double sultone has been completely consumed , the crude product is purified by semi - preparative hplc ( system b ). the mass analyses carried out have made it possible to isolate the correct fractions . a treatment on a dowex resin ( dowex ® 50wx8 - 400 ) is then necessary in order to reprotonate the sulphonate functions . a last purification on a semi - preparative column is then necessary in order to separate the cleavage product from the boc group due to the acid treatment via the resin . the final product is obtained in the form of a white solid ( 2 . 8 mg , 7 . 3 %). 1 h nmr ( dmso , 300 mhz ) δ 8 . 20 - 8 . 17 ( dd , 2h ), 7 . 82 - 7 . 77 ( t , j = hz , 2h ), 4 . 93 - 4 . 91 ( m , 1h ), 4 . 51 - 4 . 42 ( m , 1h ), 4 . 24 ( t , j = 8 . 6 hz , 2h ), 3 . 77 ( t , j = 7 . 4 hz , 2h ), 2 . 67 - 2 . 60 ( m , 2h ), 1 . 77 - 1 . 74 ( m , 2h ), 1 . 65 - 1 . 55 ( m , 2h ), 1 . 37 ( s , 9h ), 1 . 31 - 1 . 24 ( m , 2h ). hplc ( system c ): t r = 15 . 8 ( dial )− 16 ( dia2 ) min ( purity 97 . 3 %); 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