Patent Application
20140039074
The invention generally relates to the preparation of 18F-labeled radiopharmaceuticals. In particular, this invention relates to the advanced processes for an efficient elution of [18F]fluoride trapped in a cartridge filled with a quaternary ammonium polymer which comprises inert non-basic and non-nucleophilic counter anions. The said methods and polymer cartridges allow the rapid preparation of a suitable [18F]fluoride solution which is also less basic to reduce the formation of byproducts, finally to increase the radiochemical yield and purity of 18F-radiopharmaceuticals.
18F-labeled radiopharmaceuticals, tertiary alcohols, quaternary ammonium polymer, eluting solution, rapid drying,
The invention aims to prepare 18F-labeled radiopharmacueticals in high radiochemical yield and purity through the rapid process of separation/elution of [18F]fluoride ion by using an inert quaternary ammonium polymer cartridge and a volatile eluting solution.
Positron emission tomography (PET) is an emerging technology to image and diagnose numerous human diseases at an early stage. [P. W. Miller, N. J. Long, R. Vilar, A. D. Gee, Angew, Chem. Int, Ed, 2008, 47, 8998-9033] Of several positron-emitting radionuclides produced from a cyclotron, [18F]fluoride is thought to have the most suitable chemical and physical properties for diagnostic radiopharmaceuticals. The atomic size of fluorine is similar to hydrogen and the fluorine offers improved lipophilicity to fluorine-containing compounds as well as inertness to metabolic transformations.
[18F]Fluoride can be readily prepared from medical cyclotron, and has a proper half-life of about 110 min. [M. C. Lasne, C. Perrio, J. Rouden, L. Barre, D, Roeda, F. Dolle, C. Crouzel, Contrast Agents II, Topics in Current Chemistry, Springer-Verlag, Berlin, 2002, 222, 201-258.; R. Bolton, J. Labelled Compd. Radiapharm, 2002, 45 485-528].
Commonly, [18F]fluoride produced from the cyclotron exists in a highly diluted enriched O-18 water solution. [M. R. Kilbourn, J. T. Hood, M. J. Welch, Int. J. Appl. Radiat. Isot. 1984, 35, 599.; G. K, Mulholland, R. D, Hichwa, M. R. Kilbourn, J. Moskwa, J. Label, Compd. Radiopharm. 1989, 26, 140] Enriched O-18 water is very expensive and contains trace amount of metal cations after irradiation, which may influence the 18F-labeling reaction.
Some cartridges containing an anion-exchange resin are usually utilized to separate [18F]fluoride from enriched O-18 water and remove trace metal cations by solid phase extraction, [K.—I, Nishijima, Y. Kuge, E, Tsukamoto, K, —I, Seki, K, Ohkura, Y. Magaia, A, Tanaka, K. Nagatsu, N. Tamaki. Appl. Radiat. Isot. 2002, 57, 43.; D. Schoeller, Obes. Res. 1999, 7, 519.; SNM Newsline, J. Nucl. Med, 1991, 32, 15N; D. J. Schlyer, M. Bastos, A. P. Wolf, J. Nucl. Med. 1987, 28, 764.; S. A. Toorongian, G. K. Mulholland, D. M. Jewett, M. A. Bachelor, M. R. Kilbourn, Nucl. Med. Biol. 1990, 17, 273.; D. M. Jewett, S. A. Toorongian, G. K. Mulholland, G. L. Watkins, M. R, Kilbourn, Appl, Radiat. Isot. 1988, 39, 1109,; G. K. Mulholland, R. D. T. J. Mangner, D. M. Jewett, M. R. Kilbourn, J. Label. Compd. Radiopharm. 1989, 26, 378.; K. Ohsaki, Y. Endo, S. Yamazaki, M. Tomoi, R. Iwata, Appl. Radiat. Isot. 1998, 49, 373-378.]
Chromafix® and QMA cartridges are routinely used in automated radiolabeling as well as manual synthesis, and commercially available. They comprise bicarbonate and chloride counter anions, respectively. These anions possess somewhat basic and nucleophilic properties so that they may cause stability problems in long term storage. In other words, these basic anions can attack internally labile benzyl carbon atoms, resulting in free volatile tertiary amines.
To activate QMA cartridges, the chloride counter anions are exchanged with carbonate anions by eluting aqueous potassium carbonate solution before use. After the respective activation process, both Chromafix® and QMA have enough basic anions inside of the cartridge for the nucleophilic [18F]fluorination reaction. In addition, excess potassium carbonate in aqueous solution is usually used for complete release of [18F]fluoride out of these cartridges. The final [18F]fluoride solution after elution contains excess base and water.
Excess base may cause numerous side reactions including elimination and hydroxylation. Such byproduct analogues result in difficult purification of desired 18F-labeled product and low specific activity.
Large amounts of water are needed to be removed using repeated azeotropic evaporation with acetonitrile to make the reactive anhydrous [18F]fluoride ion. Protic solvents including water are known to diminish the nucleophilicity of [18F]-fluoride by building a strong hydrogen bond with [18F]fluoride. Complete evaporation requires 15-20 min, consuming 8-12% radioactivity of [18F]fluoride. This tedious evaporation process also plays a critical role in low and fluctuated reproducibility of both manual and automated synthesis.
A pioneering attempt related to [18F]-fluorination is disclosed in [J. W. Seo, E. P. Hong, B. S. Lee, S. J. Lee, S. J. Oh, D. Y. Chi, J. Labelled Compd. Radiopharm. 2007, 50 (Suppl. 1), S164], wherein a volatile alcoholic solution containing neutral ammonium-based organic salts is used to elute [18F]-fluoride trapped in polymer cartridge, resulting in great reduction of the drying time up to 1-2 min and significant suppression of side reactions.
However, neutral ammonium salts may make HPLC purification difficult by contaminating the HPLC column. That method is, therefore, limited only to the manual radiolabeling with small radioactivity. This practical restriction illustrates the need for a further advanced method suitable for the automated synthetic system.
In the invention described herein, nucleophilic [18F]fluorination is performed using tertiary alcohol solvents to avoid the formation of byproducts according to the state of the art. [D. W. Kim, D.-S. Ahn, Y—H. Oh, S. Lee, H. S. Kil, S. J. Oh, S. J. Lee, J. S. Kim, J.-S. Ryu, D. H. Moon, D. Y. Chi, J. Am. Chem. Soc. 2006, 126, 16394-16397.; D. H. Moon, D. Y. Chi, D. W. Kim, S. J. Oh, J.-S. Ryu. PCT, WO 2006/065038 A1]
The invention relates to pretreatment of [18F]fluoride for an efficient nucleophilic [18F]fluorination reaction.
This invention provides a stable neutral ionic polymer.
This invention further provides a method for the synthesis of the neutral ionic polymer.
This invention provides a cartridge by filling with the said ionic polymer.
This invention further provides a method for the separation of [18F]fluoride from enriched 0-18 water.
This invention provides volatile solutions to release [18F]fluoride trapped in the said cartridge.
This invention further provides a method to formulate the volatile eluting solution.
This invention provides a method to release [18F]fluoride trapped in the said cartridge using the said eluting solution.
This invention further provides a method to reduce the evaporation time using the said cartridge and eluting solution.
This invention provides a method to increase the radiochemical yield (RCY) of the nucleophilic [18F]fluorination by reducing the evaporation time.
This invention further provides a method to increase the RCY of the nucleophilic [18F]fluorination by using less basic said eluting solution.
This invention provides a method to decrease the amount of precursor for the ease of purification by decreasing the basicity of the nucleophilic [18F]fluorination condition.
The present invention generally relates to nucleophilic [18F]fluorination, which takes place in liquid reaction media. As shown in
The detailed present invention is described below.
In the text of the present invention, a series of quaternary ammonium polymer as illustrated in Formula 1.
Wherein R is selected from the group consisting of C1-C4 alkyl chains; 5-membered or 6-membered heterocyclic compounds having a nitrogen atom;
X is an inert alkylsulfonate or perfluoride ion having no nucleophilicity; polystyrene is the copolymer consisted of styrene, styrene derivative, and divinylbenzene (DVB).
More detailed,
NR3 is selected from the group consisting of trimethylamine, triethylamine, tri-n-propylamine, tri-n-butylamine, N-methylimidazole, and pyridine;
X is selected from the group consisting of methanesulfonate (OMs), trifluoromethanesulfonate (OTf), para-toluenesulfonate (OTs), para-nitrobenzenesulfonate (ONs), tekrafluoroborate (BF4), hexafluorophosphate (PF6), hexafluoroantimonate (SbF6), and N,N-bis(trifluoromethanesulfonyl)imide (N(Tf)2);
polystyrene is an insoluble copolymer consisting of styrene and styrene derivative, which are cross-linked with 10-90 v/v % of divinylbenzene.
In embodiments of the present invention, the said polymer may be prepared by two synthetic pathways as shown in Scheme 1.
(Wherein NR3 and polystyrene are defined above)
The first pathway (upper arrow) comprises tandem two steps. The 4-Vinylbenzyl ammonium chloride (3) intermediate is synthesized by the reaction of 4-vinylbenzyl chloride (2) and excess tertiary amine as defined above (step 1). VVithout purification, the intermediate 3 is in situ polymerized with divinylbenzene crosslinker initiated by azobisisobutyronitrile (AIBN) to give the solid polystyrene 5 (step 2). In the first step, the reaction media is selected from the group consisting of THF, CCl4, CHCl3, 1,2-dichloroethane, acetonitrile, DMF. DMSO, and water. The mixed solvent of water and DMF is proper as reaction media. The reaction in step 1 is performed at 50° C. for 3-12 h. In the second step, the reaction is performed at 70° C. for 3-12 h.
The second pathway (low arrow) comprises two separate steps. 4-Vinylbenzyl chloride (2) is polymerized with DVB crosslinker initiated by AIBN to give solid polystyrene 4, which is purified by washing and solid phase extraction using a Soxhlet apparatus (step 3). The ammonium chloride polymer 5 is prepared by quaternization of polymer 4 with excess tertiary amine as defined above (step 4).
In step 3, the reaction media is selected from the group consisting of THF, CCl4, CHCl3, 1,2-dichloroeihane, monochlorobenzene, acetonikrile, DMF. DMSO, and water. Monochlorobenzene or DMF is suitable as reaction media. The reaction in step 3 is performed at 70° C. for 3-12 h. In step 4, the reaction media is selected from the group consisting of THF, CCl4, CHCl3, 1,2-dichloroethane, acetonitrile, DMF. DMSO, and water. The mixed solvent of water and DMF is proper as the reaction media. The reaction in step 4 is performed at 70° C. for 3-24 h.
In embodiments of the present invention, the ammonium chloride polymer 5 is treated with aqueous MX solution for anion exchange from chloride to the inert X anion as shown in Scheme 2.
(Wherein NR3. X, and polystyrene are defined above)
In Scheme 2, M is selected from the group consisting of lithium (Li), sodium (Na), potassium (K), 1-n-butyl-3-methylimidazolium ([bmim]), pyridinium, substituted pyridinium, phosphonium, and NR4 (R=Me, Et, n-Pr, n-Bu). The anion exchanging process is carried out as follows;
In embodiments of the present invention, the said polymer 1 is used to make a more stable and efficient solid phase extraction cartridge to separate [18F]fluoride and to prepare a less basic [18F]fluoride solution.
For complete releasing [18F]fluoride out of the said cartridge and fast evaporation, an effective eluting solution is prepared by composing K222, KOMs (or KOTf, K3PO4), and TBAHCO3 (or TBAOH, or KOH, or K2CO3. or KHCO3). Wherein K222 is the most effective phase transfer catalyst in nucleophilic [18F]fluorination; KOMs and KOTf are the source of inert anion instead of TBAOMs disclosed in KP application #10-2008-0078233 for complete solid phase extraction of [18F]fluoride; K3PO4, TBAHCO3, TBAOH, KOH, K2CO3, and KHCO3 are used to keep reaction solution basic. These components are diluted in an alcohol solvent which is selected form the group consisting of primary alcohol such as methanol, ethanol, n-propanol, n-butanol, n-pentanol, n-hexanol, n-heptanol, and n-octanol; or sencondary alcohol such as isopropanol, isobutanol, isoamyl alcohol, and 3-pentanol; or tertiary alcohol such as t-butanol, t-amyl alcohol, 2,3-dimethyl-2-butanol, 2-(trifluoromethyl)-2-propanol, 3-methyl-3-pentanol, 3-ethyl-3-pentanol, 2-emthyl-2-pentanol, 2,3-dimethyl-3-pentanol, 2,4-dimethyl-2-pentanol, 2-methyl-2-hexanol, 2-cyclopropyl-2-propanol, 2-cyclopropyl-2-butanol, 2-cyclopropyl-3-methyl-2-butanol, 1-methylcyclopentanol, 1-ethylcyclopentanol, 1-propylcyclopentanol, 1-methylcyclohexanol, 1-ethylcyclohexanol, 1-methylcycloheptanol.
In embodiments of the present invention, the eluted [18F]fluoride solution out of the said polymer cartridge is evaporated under a gentle now of N2 or He gas and low vacuum. The small amount of water is then removed by azeotropic evaporation with acetonitrile under a gentle flow of N2 or He gas and low vacuum.
The content of the present invention is not limited to EXAMPLES below.
After dissolving 4-vinylbenzyl chloride (2, 1.00 mL, 7.096 mmol) in a mixed solution of water (0.5 mL) and DMF (5.0 mL), 40% trimethylamine aqueous solution (2.098 mL, 14.190 mmol) was added to the solution. The reaction mixture was stirred at 50° C. for 3 h to give N-(4-vinylbenzyl)trimethylammonium chloride (3-1) (step 1). After cooling to room temperature, divinylbenzene (2.00 mL, 11.233 mmol) and AIBN (301 mg, 1.833 mmol) were added and dissolved completely. The reaction mixture was heated at 70° C. for 5 h, and then cooled to room temperature. The resulting polymeric solid (5-1) was roughly crushed and transferred into a 400 mesh sieve, and then was washed with acetone several times (step 2). After drying the polymeric solid under atmosphere, it was grinded in a mortar to result in small particles, and then sorted by particle size using stacked four different sieves to give trimethylammonium chloride polystyrene (5-1); 50-100 mesh: 2.25 g, 100-200 mesh: 0.248 g, 200-400 mesh 0.208 g.
Using triethylairne (1.978 mL, 14.190 mmol) instead of trimethylamine of example 1 above, and following the same procedure and reaction scale as example 1, triethylammonium chloride polystyrene (5-2) was obtained as follows; 50-100 mesh: 2.374 g, 100-200 mesh: 0.487 g, 200-400 mesh: 0.221 g.
Using N-methylimidazole (1.131 mL, 14.190 mmol) instead of trimethylamine of example 1 above, and following the same procedure and reaction scale as example 1. N-methylimidazolium chloride polystyrene (5-3) was obtained as follows; 50-100 mesh: 1.120 g. 100-200 mesh: 1.377 g, 200-400 mesh: 0.189 g.
Using pyridine (1.148 mL, 14.190 mmol) instead of trimethylamine of example 1 above, and following the same procedure and reaction scale as example 1, pyrimidinium chloride polystyrene (5-4) was obtained as follows; 50-100 mesh: 1.719 g, 100-200 mesh: 0.206 g, 200-400 mesh: 0.582 g.
An elemental analysis of four ammonium chloride polymers obtained from above examples 1-4 was obtained and the amount of ammonium ion of resins was calculated on the basis of nitrogen content (%) as shown in Table 1,
Polymer 5-1 (100-200 mesh, 200 mg) obtained from example 1 was placed into a syringe equipped with a polyethylene frit. Distilled water (10 mL) was added into the syringe and eluted out after 1 min. The syringe was flushed with 0.2 M NaOMs aqueous solution (5 mL) and capped with a tight lid, and then shaked for 3 min. The solution was removed by filtration under reduced pressure and the resin was washed with distilled water. After the ion exchange process was repeated four times, the resin was washed with distilled water (5 mL×5) and acetone (5 mL×5) and then dried under vacuum to give the trimethylammonium methanesulfonate polystyrene (1-1, 235 mg).
From polymer 5-2 (100-200 mesh, 200 mg), triethylammonium methanesulfonate polystyrene (1-2, 222 mg) was prepared by following the same procedure as example 5.
From polymer 5-3 00-200 mesh, 200 mg), N-methylimidazolium methanesulfonate polystyrene (1-3, 225 mg) was prepared by following the same procedure as example 5.
From polymer 5-4 (100-200 mesh, 200 mg), N-methylimidazolium methanesulfonate polystyrene (1-4, 220 mg) was prepared by following the same procedure as example 5.
The neutral ammonium methanesulfonate polymers 1 ranging from 20 mg to 100 mg were filled into a cartridge equipped with a polyethylene frit.
Polymer cartridge 6-1 were prepared by being filled with polymer 1-1
Polymer cartridge 6-2 were prepared by being filled with polymer 1-2
Polymer cartridge 6-3 were prepared by being filled with polymer 1-3
Polymer cartridge 6-4 were prepared by being filled with polymer 1-4
The eluting solutions for releasing [18F]fluoride captured in a cartridge were prepared by composing three ingredients, and dissolved in alcohol solvent.
Ingredient B: 0.05-0.2 M KOMs, KOTf, K3PO4 in water; 0.05-0.2 mL
Each ingredient was selected from each group A, B. and C. and mixed together to make several eluting solutions as follows;
2) 0.2 M KOMs in water; 0.05-0.2 mL
4) alcohol; 1 mL
2) 0.2 M KOTf in water; 0.05-0.2 mL
4) alcohol; 1 mL
2) 0.2 M K3PO4 in water; 0.05-0.2 mL
4) alcohol; 1 mL
2) 0.2 M KOMs in water; 0.05-0.2 mL
4) alcohol; 1 mL
2) 0.2 M KOMs in water; 0.05-0.2 mL
3) 0.05-0.2 M KOH in water; 0.01-0.2 mL
4) alcohol; 1 mL
2) 0.2 M KOMs in water; 0.05-0.2 mL
3) 0.05-0.2 M K2CO3 in water; 0.01-0.2 mL
4) alcohol; 1 mL
2) 0.2 M KOMs in water; 0.05-0.2 mL
3) 0.05-0.2 M KHCO3 in water; 0.01-0.2 mL
4) alcohol; 1 mL
Dilute aqueous [18]fluoride solution (ca. 3-6 mCi) was passed through the cartridges (6-1-6-4) prepared by present invention to trap [18F]fluoride. The [18F]fluoride-trapped cartridge was then washed with distilled water (1.0 mL) and methanol solvent (1.0 mL) in sequence. [18F]Fluoride trapped in the cartridge was released by eluting the solution (Eluent A) prepared in the present invention. The released amount of [18F]fluoride from the cartridge was counted every 0.1 mL elution. The result of elution using present invention is summarized in Table 3.
Aqueous [18F]fluoride solution was passed through the cartridge (6-3) of the present invention. No [18F]fluoride was detected in the filtrate solution and 92.1-115.4 MBq of [18F]fluoride was trapped in the cartridge. The trapped [18F]fluoride was eluted with the Eluent (A, B, or C) solution of the present invention into a reaction vial, Remained radioactivity in the cartridge was 1.85˜2.96 MBq. The eluted solution was heated at 100° C. with a gentle flow of N2 gas to remove volatile solvent, and then acetonitrile (0.5 mL) was added to the reaction vial. Azeotropic evaporation was repeated. Complete removal of solvent including water took in a range from 1 min and 30 seconds to 2 min. A solution of precursor (5 mg) dissolved in a co-solvent of t-amyl alcohol (1.0 mL) and acetonitrile (0.1 mL) was added to the reaction vial. The reaction mixture was heated at 100° C. for 10 min, and then cooled to room temperature. Radio-TLC scanning showed 90.9% of radiolabeling.
Aqueous [18F]fluoride solution was passed through the cartridge (6-3) of the present invention, No [18F]fluoride was detected in the filtrate solution and 195.4 MBq of [18F]fluoride was trapped in the cartridge. The trapped [18F]fluoride was eluted with the Eluent A solution of the present invention into a reaction vial. Remained radioactivity in the cartridge was 11.47 MBq. The eluted solution was heated at 100° C. with a gentle flow of N2 gas to remove volatile solvent, and then acetonitrile (0.5 mL) was added to the reaction vial. Azeotropic evaporation was repeated. Complete removal of solvent including water took in a range from 1 min and 30 seconds to 2 min. A solution of precursor (4 mg) dissolved in a co-solvent of t-amyl alcohol (1.0 mL) and acetonitrile (0.05 mL) was added to the reaction vial. The reaction mixture was heated at 100° C. for 20 min, and then cooled to room temperature. Radio-TLC scanning showed 86.8% of radiolabeling. HPLC purification (Varian, Bondclon C18 column 250 mm×10 mm, H2O: EtOH:Et3N=250:750:2, 4 mL/min, at 229 nm) was performed to give [18]FP-CIT in 67.9% of radiochemical yield (RCY, decay-corrected). Total preparation including HPLC purification spent 50 min.
Aqueous [18F]fluoride solution was passed through the cartridge (6-3) of the present invention. No [18F]fluoride was detected in the filtrate solution and 356.3 MBq of [18F]fluoride was trapped in the cartridge. The trapped [18F]fluoride was eluted with the Eluent D solution of the present invention into a reaction vial. Remained radioactivity in the cartridge was 54.8 MBq. The eluted solution was heated at 100° C. with a gentle flow of N2 gas to remove volatile solvent, and then acetonitrile (0.5 mL) was added to the reaction vial. Azeotropic evaporation was repeated. Complete removal of solvent including water took in a range from 1 min and 30 seconds to 2 min. A solution of precursor (4 mg) dissolved in a co-solvent of t-amyl alcohol (1.0 mL) and acetonitrile (0.05 mL) was added to the reaction vial. The reaction mixture was heated at 120° C. for 20 min, and then cooled to room temperature. Radio-TLC scanning showed 52.2% of radiolabeling. HPLC purification was performed to give [18F]FP-CIT in 42.4% of radiochemical yield (RCY, decay-corrected). Total preparation including HPLC purification spent. 50 min.
Aqueous [18F]fluoride solution was passed through the cartridge (6-3) of the present invention. No [18F]fluoride was detected in the filtrate solution and 207.9 MBq of [18F]fluoride was trapped in the cartridge. The trapped [18F]fluoride was eluted with the Eluent E solution of the present invention into a reaction vial. Remained radioactivity in the cartridge was 9.25 MBq. The eluted solution was heated at 100° C. with a gentle flow of N2 gas to remove volatile solvent, and then acetonitrile (0.5 mL) was added to the reaction vial. Azeotropic evaporation was repeated. Complete removal of solvent including water took in a range from 1 min and 30 seconds to 2 min. A solution of precursor (4 mg) dissolved in a co-solvent of t-amyl alcohol (1.0 mL) and acetonitrile (0.05 mL) was added to the reaction vial. The reaction mixture was heated at 100° C. for 20 min, and then cooled to room temperature. Radio-TLC scanning showed 95.1% of radiolabeling. HPLC purification was performed to give [18F]FP-CIT in 49.5% of radiochemical yield (RCY, decay-corrected). Total preparation including HPLC purification spent 51 min.
Aqueous [18F]fluoride solution was passed through the cartridge (6-3) of the present invention. No [18F]fluoride was detected in the filtrate solution and 147.9 MBq of [18F]fluoride was trapped in the cartridge. The trapped [18F]fluoride was eluted with the Eluent F solution of the present invention into a reaction vial. Remained radioactivity in the cartridge was 1.25 MBq. The eluted solution was heated at 100° C. with a gentle flow of N2 gas to remove volatile solvent, and then acetonitrile (0.5 mL) was added to the reaction vial. Azeotropic evaporation was repeated. Complete removal of solvent including water took in a range from 1 min and 30 seconds to 2 min. A solution of precursor (4 mg) dissolved in a co-solvent of t-amyl alcohol (1.0 mL) and acetonitrile (0.05 mL) was added to the reaction vial. The reaction mixture was heated at 100° C. for 20 min, and then cooled to room temperature. Radio-TLC scanning showed 53.6% of radiolabeling.
Aqueous [18F]fluoride solution was passed through the cartridge (6-3) of the present invention. No [18F]fluoride was detected in the filtrate solution and 214.49 MBq of [18F]fluoride was trapped in the cartridge. The trapped [18F]fluoride was eluted with the Eluent A solution of the present invention into a reaction vial. Remained radioactivity in the cartridge was 61.5 MBq. The eluted solution was heated at 100° C. with a gentle flow of N2 gas to remove volatile solvent, and then acetonitrile (0.5 mL) was added to the reaction vial. Azeotropic evaporation was repeated. Complete removal of solvent including water took in a range from 1 min and 30 seconds to 2 min. A solution of precursor (20 mg) dissolved in a co-solvent. of t-amyl alcohol (1.0 mL) and acetonitrile (0.1 mL) was added to the reaction vial. The reaction mixture was heated at 100° C. for 20 min, and then cooled to room temperature. Radio-TLC scanning showed 92.8% of radiolabeling. The solvent was removed by N2 purging under heat. at 100° C. The residue was dissolved in acetonitrile (0.5 mL), and than diluted with water (20 mL). The diluted solution was passed through a C18 SepPak cartridge, which and then filled with 2 M aqueous NaOH solution (1 mL), and left for 2 min at room temperature for hydrolysis. The reaction mixture was passed through IC-H cartridge and almunia N SepPak cartridge in sequence to give 2-[18F]fluoro-deoxyglucose ([18F]FDG) in 61.9% of RCY (decay-corrected). Total preparation including HPLC purification spent 50 min.
Aqueous [18F]fluoride solution was passed through the cartridge (6-3) of the present invention. No [18F]fluoride was detected in the filtrate solution and 148.0 MBq of [18F]fluoride was trapped in the cartridge. The trapped [18F]fluoride was eluted with the Eluent E solution of the present invention into a reaction vial. Remained radioactivity in the cartridge was 9.25 MBq. The eluted solution was heated at 100° C. with a gentle flow of N2 gas to remove volatile solvent, and then acetonitrile (0.5 mL) was added to the reaction vial. Azeotropic evaporation was repeated. Complete removal of solvent including water took in a range from 1 min and 30 seconds to 2 min. A solution of precursor (5 mg) dissolved in a co-solvent of t-amyl alcohol (1.0 mL) and acetonitrile (0.1 mL) was added to the reaction vial. The reaction mixture was heated at 120° C. for 10 min, and then cooled to room temperature. Radio-TLC scanning showed 77.7% of radiolabeling. The solvent was removed by N2 purging under heat at 100′C. The residue was dissolved in acetonitrile (0.5 mL), and then diluted with water (20 mL). The diluted solution was passed through a C18 SepPak cartridge, which and then filled with 2 M aqueous NaOH solution (1 mL), and left for 2 min at room temperature for hydrolysis. The reaction mixture was passed through IC-H cartridge and almunia N SepPak cartridge in sequence to give 2-[18F]fluoro-deoxyglucose ([18F]FDG) in 48.9% of RCY (decay-corrected). Total preparation including HPLC purification spent 42 min.
Aqueous [18F]fluoride solution was passed through the cartridge (6-3) of the present invention. No [18F]fluoride was detected in the filtrate solution and 192.3 MBq of [18F]fluoride was trapped in the cartridge. The trapped [18F]fluoride was eluted with the Eluent A solution of the present invention to a reaction vial. Remained radioactivity in the cartridge was 15.2 MBq. The eluted solution was heated at 120° C. with a gentle flow of N2 gas to remove volatile solvent, and then acetonitrile (0.5 mL) was added to the reaction vial. Azeotropic evaporation was repeated. Complete removal of solvent including water took in a range from 1 min and 30 seconds to 2 min. A solution of precursor (20 mg) dissolved in a co-solvent of t-amyl alcohol (1.0 mL) and acetonitrile (0.1 mL) was added to the reaction vial. The reaction mixture was heated at 120° C. for 20 min, and then cooled to room temperature. Radio-TLC scanning showed 93.3% of radiolabeling. The solvent was removed by N2 purging under heat at 100° C. The residue was dissolved in acetonitrile (0.1 mL) and diluted with 1 M HCl aqueous solution (0.5 mL). The solution was heated at 85° C. for 5 min, and then treated with 2 M NaOH aqueous solution (0.25 mL). HPLC purification (TSP, Econosil C18 column 250 mm×10 mm, H2O:EtOH=90:10, 5 mL/min. at 267 nm) was performed to give [18F]FLT in 48.6% of radiochemical yield (RCY, decay-corrected). Total preparation including HPLC purification spent 55 min.
Aqueous [18F]fluoride solution was passed through the cartridge (6-3) of the present invention. No [18F]fluoride was detected in the filtrate solution and 212.7 MBq of [18F]fluoride was trapped in the cartridge. The trapped [18F]fluoride was eluted with the Eluent E solution of the present invention to a reaction vial. Remained radioactivity in the cartridge was 16.3 MBq. The eluted solution was heated at 120° C. with a gentle flow of N2 gas to remove volatile solvent, and then acetonitrile (0.5 mL) was added to the reaction vial. Azeotropic evaporation was repeated. Complete removal of solvent including water took in a range from 1 min and 30 seconds to 2 min. A solution of precursor (20 mg) dissolved in a co-solvent of t-amyl alcohol (1.0 mL) and acetonitrile (0.1 mL) was added to the reaction vial, The reaction mixture was heated at 100° C. for 10 min, and then cooled to room temperature. Radio-TLC scanning showed 74.8% of radiolabeling.
Aqueous [18F]fluoride solution was passed through the cartridge (6-3) of the present invention. No [18F]fluoride was detected in the filtrate solution and 375.1 MBq of [18F]fluoride was trapped in the cartridge. The trapped [18F]fluoride was eluted with the Eluent G solution of the present invention to a reaction vial. Remained radioactivity in the cartridge was 27.9 MBq. The eluted solution was heated at 120° C. with a gentle flow of N2 gas to remove volatile solvent, and then acetonitrile (0.5 mL) was added to the reaction vial. Azeotropic evaporation was repeated. Complete removal of solvent including water took in a range from 1 min and 30 seconds to 2 min. A solution of precursor (10 mg) dissolved in a co-solvent of t-amyl alcohol (1.0 mL) and acetonitrile (0.1 mL) was added to the reaction vial. The reaction mixture was heated at 100° C. for 20 min, and then cooled to room temperature. Radio-TLC scanning showed 77.5% of radiolabeling.
Aqueous [18F]fluoride solution was passed through the cartridge (6-3) of the present invention. No [18F]fluoride was detected in the filtrate solution and 145.9 MBq of [18F]fluoride was trapped in the cartridge. The trapped [18F]fluoride was eluted with the Eluent A solution of the present invention to a reaction vial. Remained radioactivity in the cartridge was 12.4 MBq. The eluted solution was heated at 120° C. with a gentle flow of N2 gas to remove volatile solvent, and then acetonitrile (0.5 mL) was added to the reaction vial. Azeotropic evaporation was repeated. Complete removal of solvent including water took in a range from 1 min and 30 seconds to 2 min. A solution of precursor (10 mg) dissolved in a co-solvent of t-amyl alcohol (1.0 mL) and acetonitrile (0.1 mL) was added to the reaction vial. The reaction mixture was heated at 120° C. for 10 min, and then cooled to room temperature. Radio-TLC scanning showed 96.1% of radiolabeling. The solvent was removed by N2 purging under heat at 100° C. The residue was dissolved in acetonitrile (0.1 mL) and diluted with 1 M HCl aqueous solution (0.5 mL). The solution was heated at 85° C. for 5 min, and then treated with 2 M NaOH aqueous solution (0.25 mL). HPLC purification (TSP, Econosil C18 column 250 mm×10 mm H2O:EtOH=95:5, 5 mL/min, at 254 nm) was performed to give [18]FMISO in 42.3% of radiochemical yield (RCY, decay-corrected). Total preparation including HPLC purification spent 45 min.
Aqueous [18F]fluoride solution was passed through the cartridge (6-3) of the present invention. No [18F]fluoride was detected in the filtrate solution and 294.2 MBq of [18F]fluoride was trapped in the cartridge. The trapped [18F]fluoride was eluted with the Eluent A solution of the present invention to a reaction vial. Remained radioactivity in the cartridge was 35.5 MBq. The eluted solution was heated at 120° C. with a gentle flow of N2 gas to remove volatile solvent, and then acetonitrile (0.5 mL) was added to the reaction vial, Azeotropic evaporation was repeated. Complete removal of solvent including water took in a range from 1 min and 30 seconds to 2 min. A solution of precursor (4 mg) dissolved in a co-solvent of t-amyl alcohol (1.0 mL) and acetonitrile (0.1 mL) was added to the reaction vial. The reaction mixture was heated at 120° C. for 10 min, and then cooled to room temperature. Radio-TLC scanning showed 81.1% of radiolabeling. The solvent was removed by N2 purging under heat at 120° C. The residue was dissolved in acetonitrile (0.3 mL) and diluted with 1 M HCl aqueous solution (0.5 mL). The solution was heated at 120° C. for 5 min, and then treated with 2 M NaOH aqueous solution (0.25 mL), HPLC purification (Varian, Gemini C18 column 250 mm×10 mm 0.1 M ammonium formate:MeCN=40:60, 4 mL/min, at 254 nm) was performed to give [18F]BAY94-9172 in 58.1% of radiochemical yield (RCY, decay-corrected). Total preparation including HPLC purification spent 60 min.
Aqueous [18F]fluoride solution was passed through the cartridge (6-3) of the present invention. No [18F]fluoride was detected in the filtrate solution and 154.3 MBq of [18F]fluoride was trapped in the cartridge. The trapped [18F]fluoride was eluted with the Eluent D solution of the present invention to a reaction vial. Remained radioactivity in the cartridge was 13.0 MBq. The eluted solution was heated at 120° C. with a gentle flow of N2 gas to remove volatile solvent, and then acetonitrile (0.5 mL) was added to the reaction vial. Azeotropic evaporation was repeated. Complete removal of solvent including water took in a range from 1 min and 30 seconds to 2 min. A solution of precursor (4 mg) dissolved in a co-solvent of t-amyl alcohol (1.0 mL) and acetonitrile (0.1 mL) was added to the reaction vial. The reaction mixture was heated at 120° C. for 20 min, and then cooled to room temperature. Radio-TLC scanning showed 86.91% of radiolabeling. The solvent was removed by N2 purging under heat at 120° C. The residue was dissolved in acetonitrile (0.3 mL) and diluted with 1 M HCl aqueous solution (0.5 mL). The solution was heated at 120° C. for 5 min, and then treated with 2 M NaOH aqueous solution (0.25 mL). HPLC purification (Varian, Gemini C18 column 250 mm×10 mm 0.1 M ammonium formate:MeCN=40:60, 4 mL/min. at 254 nm) was performed to give [18F]BAY94-9172 in 68.9% of radiochemical yield (RCY, decay-corrected). Total preparation including HPLC purification spent 60 min.
Aqueous [18F]fluoride solution was passed through the cartridge (6-3) of the present invention. No [18F]fluoride was detected in the filtrate solution and 173.2 MBq of [18F]fluoride was trapped in the cartridge. The trapped [18F]fluoride was eluted with the Eluent G solution of the present invention to a reaction vial. Remained radioactivity in the cartridge was 1.48 MBq. The eluted solution was heated at 120° C. with a gentle flow of N2 gas to remove volatile solvent, and then acetonitrile (0.5 mL) was added to the reaction vial. Azeotropic evaporation was repeated. Complete removal of solvent including water took in a range from 1 min and 30 seconds to 2 min. A solution of precursor (4 mg) dissolved in a co-solvent of t-amyl alcohol (1.0 mL) and acetonitrile (0.1 mL) was added to the reaction vial. The reaction mixture was heated at 120° C. for 20 min, and then cooled to room temperature. Radio-TLC scanning showed 86.9% of radiolabeling. The solvent was removed by N2 purging under heat at 120° C. The residue was dissolved in acetonitrile (0.3 mL) and diluted with 1 M HCl aqueous solution (0.5 mL). The solution was heated at 120° C. for 5 min, and then treated with 2 M NaOH aqueous solution (0.25 mL). HPLC purification was performed to give [18]BAY94-9172 in 52.2% of radiochemical yield (RCY, decay-corrected). Total preparation including HPLC purification spent 60 min.
Aqueous [18F]fluoride solution was passed through the cartridge (6-3) of the present invention. No [18F]fluoride was detected in the filtrate solution and 330.8 GBq of [18F]fluoride was trapped in the cartridge. The trapped [18F]fluoride was eluted with the Eluent A solution of the present invention to a reaction vial. Remained radioactivity in the cartridge was 43.3 MBq. The eluted solution was heated at 120° C. with a gentle flow of N2 gas to remove volatile solvent, and then acetonitrile (0.5 mL) was added to the reaction vial. Azeotropic evaporation was repeated. Complete removal of solvent including water took in a range from 1 min and 30 seconds to 2 min. A solution of precursor (2 mg) dissolved in a co-solvent of t-amyl alcohol (1.0 mL) and acetonitrile (0.1 mL) was added to the reaction vial. The reaction mixture was heated at 100° C. for 5 min, and than cooled to room temperature. Radio-TLC scanning showed 92.4% of radiolabeling. HPLC purification (Varian, Econosil C18 column 250 mm×10 mm 50 mM triethylammonium phosphate:MeCN=40:60, 4 mL/min, at 254 nm) was performed to give [18F]FDDNP in 48.5% of radiochemical yield (ROY, decay-corrected). Total preparation including HPLC purification spent 61 min.
Aqueous [18F]fluoride solution was passed through the cartridge (6-3) of the present invention. No [18F]fluoride was detected in the filtrate solution and 259.8 GBq of [18F]fluoride was trapped in the cartridge, The trapped [18F]fluoride was eluted with the Eluent F solution of the present invention to a reaction vial. Remained radioactivity in the cartridge was 23.3 MBq. The eluted solution was heated at 120° C. with a gentle flow of N2 gas to remove volatile solvent, and then acetonitrile (0.5 mL) was added to the reaction vial. Azeotropic evaporation was repeated. Complete removal of solvent including water took in a range from 1 min and 30 seconds to 2 min. A solution of precursor (2 mg) dissolved in a co-solvent of t-amyl alcohol (1.0 mL) and acetonitrile (0.1 mL) was added to the reaction vial. The reaction mixture was heated at 100° C. for 5 min, and then cooled to room temperature. Radio-TLC scanning showed 85.1% of radiolabeling. HPLC purification (Varian. Econosil 018 column 250 mm×10 mm 50 mM triethylammonium phosphate:MeCN=40:60, 4 mL/min, at 254 nm) was performed to give [18F]FDDNP in 48.5% of radiochemical yield (RCY, decay-corrected). Total preparation including HPLC purification spent 61 min.
Aqueous [18F]fluoride solution was passed through the cartridge (6-3) of the present invention, No [18F]fluoride was detected in the filtrate solution and 210.7 GBq of [18F]fluoride was trapped in the cartridge. The trapped [18F]fluoride was eluted with the Eluent G solution of the present invention to a reaction vial. Remained radioactivity in the cartridge was 16,3 MBq. The eluted solution was heated at 120° C. with a gentle flow of N2 gas to remove volatile solvent, and then acetonitrile (0.5 mL) was added to the reaction vial. Azeotropic evaporation was repeated. Complete removal of solvent including water took in a range from 1 min and 30 seconds to 2 min. A solution of precursor (2 mg) dissolved in a co-solvent of t-amyl alcohol (1.0 mL) and acetonitrile (0.1 mL) was added to the reaction vial. The reaction mixture was heated at 100° C. for 5 min, and then cooled to room temperature. Radio-TLC scanning showed 95.9% radiolabeling (RCY, decay-corrected). Total preparation including HPLC purification spent 65 min.
Aqueous [18F]fluoride solution was passed through the cartridge (6-3) of the present invention. No [18F]fluoride was detected in the filtrate solution and 2.49 GBq of [18F]fluoride was trapped in the cartridge. The trapped [18F]fluoride was eluted with the Eluent A solution of the present invention to a reaction vial. Remained radioactivity in the cartridge was 51.8 MBq. The eluted solution was heated at 120° C. with a gentle flow of N2 gas to remove volatile solvent, and then acetonitrile (0.5 mL) was added to the reaction vial. Azeotropic evaporation was repeated. Complete removal of solvent including water took in a range from 1 min and 30 seconds to 2 min. A solution of precursor (4 mg) dissolved in a co-solvent of t-amyl alcohol (1.0 mL) and acetonitrile (0.1 mL) was added to the reaction vial. The reaction mixture was heated at 120° C. for 20 min, and then cooled to room temperature. Radio-TLC scanning showed 92.4% of radiolabeling. The solvent was removed by N2 purging under heat at 120° C. The residue was dissolved in acetonitrile (0.3 mL) and diluted with 1 M HCl aqueous solution (0.5 mL). The solution was heated at 120° C. for 5 min, and then treated with 2 M NaOH aqueous solution (0.25 mL). HPLC purification was performed to give [18]AV-45 in 59.4% of radiochemical yield (RCY, decay-corrected), Total preparation including HPLC purification spent 81 min.
In particular, the invention relates to:
| Number | Date | Country | Kind |
|---|---|---|---|
| 10009380.6 | Sep 2010 | EP | regional |
| Filing Document | Filing Date | Country | Kind | 371c Date |
|---|---|---|---|---|
| PCT/EP2011/065366 | 9/6/2011 | WO | 00 | 10/18/2013 |
