Patent Application
20140243533
This invention relates to an improved method, which provides access to [F-18]fluoropegylated (aryl/heteroaryl vinyl)-phenyl methyl amine derivatives.
Alzheimer's Disease (AD) is a progressive neurodegenerative disorder marked by loss of memory, cognition, and behavioral stability. AD is defined pathologically by extracellular senile plaques comprised of fibrillar deposits of the beta-amyloid peptide (Aβ) and neurofibrillary tangles comprised of paired helical filaments of hyperphosphorylated tau. The 39-43 amino acids comprising Aβ peptides are derived from the larger amyloid precursor protein (APP). In the amyloidogenic pathway, Aβ peptides are cleaved from APP by the sequential proteolysis by beta- and gamma-secretases. Aβ peptides are released as soluble proteins and are detected at low level in the cerebrospinal fluid (CSF) in normal aging brain. During the progress of AD the Aβ peptides aggregate and form amyloid deposits in the parenchyma and vasculature of the brain, which can be detected post mortem as diffuse and senile plaques and vascular amyloid during histological examination (for a recent review see: Blennow et al. Lancet. 2006 Jul. 29; 368(9533):387-403).
Alzheimer's disease (AD) is becoming a great health and social economical problem all over the world. There are great efforts to develop techniques and methods for the early detection and effective treatment of the disease. Currently, diagnosis of AD in an academic memory-disorders clinic setting is approximately 85-90% accurate (Petrella J R et al. Radiology. 2003 226:315-36). It is based on the exclusion of a variety of diseases causing similar symptoms and the careful neurological and psychiatric examination, as well as neuropsychological testing.
Molecular imaging has the potential to detect disease progression or therapeutic effectiveness earlier than most conventional methods in the fields of neurology, oncology and cardiology. Among the several promising molecular imaging technologies, such as optical imaging, MRI, SPECT and PET, PET is of particular interest for drug development because of its high sensitivity and ability to provide quantitative and kinetic data.
For example positron emitting isotopes include e.g. carbon, iodine, nitrogen and oxygen. These isotopes can replace their non-radioactive counterparts in target compounds to produce PET tracers that have similar biological properties. Among these isotopes F-18 is a preferred labeling isotope due to its half life of 110 min, which permits the preparation of diagnostic tracers and subsequent study of biochemical processes. In addition, its low β+ energy (634 keV) is also advantageous.
Post-mortem histological examination of the brain is still the only definite diagnosis of Alzheimer's disease. Thus, the in vivo detection of one pathological feature of the disease—the amyloid aggregate deposition in the brain—is thought to have a strong impact on the early detection of AD and differentiating it from other forms of dementia. Additionally, most disease modifying therapies which are in development are aiming at lowering of the amyloid load in the brain. Thus, imaging the amyloid load in the brain may provide an essential tool for patient stratification and treatment monitoring (for a recent review see: Nordberg. Eur J Nucl Med Mol Imaging. 2008 March; 35 Suppl 1:S46-50).
In addition, amyloid deposits are also known to play a role in amyloidoses, in which amyloid proteins (e.g. tau) are abnormally deposited in different organs and/or tissues, causing disease. For a recent review see Chiti et al. Annu Rev Biochem. 2006; 75:333-66.
Fluoropegylated (aryl/heteroaryl vinyl)-phenyl methyl amines such as 4-[(E)-2-(4-{2-[2-(2-fluoroethoxy)ethoxy]ethoxy}phenyl)vinyl]-N-methylaniline and 4-[(E)-2-(6-{2-[2-(2-fluoroethoxy)ethoxy]ethoxy}pyridin-3-yl)vinyl]-N-methylaniline have been labeled with F-18 fluoride and are covered by patent applications WO2006066104, WO2007126733 and members of the corresponding patent families.
The usefulness of these radiotracers for the detection of Aβ plaques have been reported in the literature (W. Zhang et al., Nuclear Medicine and Biology 32 (2005) 799-809; C. Rowe et al., Lancet Neurology 7 (2008) 1-7; S. R. Choi et al., The Journal of Nuclear Medicine 50 (2009) 1887-1894).
To not limit the use of such F-18 labeled diagnostics, processes are needed, that allow a robust and safe manufacturing of the F-18 labeled tracers. Additionally, such processes should provide high yield of the overall synthesis to allow the production of quantities of the diagnostic to supply the radiotracer, despite of the half life of 110 min, to facilities without cyclotron or radiopharmaceutical production facility.
Syntheses of F-18 labeled fluoropegylated (aryl/heteroaryl vinyl)-phenyl methyl amines have been described before. For most of these radiosyntheses a F-18 eluent mixture typically comprises kryptofix 2.2.2, potassium carbonate, acetonitrile and water.
The general setup of the manufacturing process for F-18 labeled fluoropegylated (aryl/heteroaryl vinyl)-phenyl methyl amines as previously described is illustrated in
During the first manufacturing step [F-18]fluoride is eluted from a ion exchange cartridge (e.g. QMA) usually using an eluent mixture comprising cryptand (e.g. kryptofix 2.2.2), a base (e.g. potassium carbonate), acetonitrile and water. The manufacturing steps are:
These steps outlined above are performed on the part A of the synthesis device (
Typically (and described in the references above), the product fraction is diluted with water (vessel “8”,
A “GMP compliant” manufacturing process for 4-[(E)-2-(6-{2-[2-(2-[F-18]fluoroethoxy)ethoxy]-ethoxy}pyridin-3-yl)vinyl]-N-methylaniline is disclosed in WO2010078370 and C.-H. Yao et al., Applied Radiation and Isotopes 68 (2010) 2293-2297. To prevent the decomposition of 4-[(E)-2-(6-{2-[2-(2-[F-18]fluoroethoxy)ethoxy]-ethoxy}pyridin-3-yl)vinyl]-N-methylaniline, sodium ascorbate was added to the HPLC solvent (45% acetonitrile, 55% 20 mM ammoniumacetate containing 0.5% (w/v) sodium ascorbate) and the final Formulation (0.5% (w/v) sodium ascorbate). The process afforded up to 18.5 GBq (25.4±7.7%, decay corrected) 4-[(E)-2-(6-{2-[2-(2-[F-18]fluoroethoxy)ethoxy]-ethoxy}pyridin-3-yl)vinyl]-N-methylaniline. The radiochemical purity was 95.3±2.2%.
Although, cartridge based purification processes have been investigated, an optimum of product quality regarding radiochemical purity and separation from (non-radioactive) by-products have been demonstrated and proofed only for HPLC purification.
Here, a new eluation method for [F-18]-fluoride in the first manufacturing step is provided.
To take full advantage of the new purification method it turned out to be benefical to start with a crude product which can be purified easily by HPLC whereby the mobile phase comprises of an ethanol/buffer mixture. Typically [F-18]fluoride is trapped on a anion exchange resin (e.g. QMA cartridge or SPE) and then eluted from this anion exchange resin with a basic solution consisting of base, acetonitrile and water and optionally a cryptand. The most commonly used solution for elution of [F-18]fluoride trapped on a anion exchange resin comprise of varying amounts of kryptofix 2.2.2. (cryptand), potassium carbonate (base), acetonitrile and water (conventional eluent mixture). However, in particular if older batches of this conventional eluent mixture or conventional eluent batches from accelerated stability studies are used, in case of the synthesis of 4-[(E)-2-(4-{2-[2-(2-fluoroethoxy)ethoxy]ethoxy}phenyl)vinyl]-N-methylaniline the crude product contains major amounts of a chemical impurity (Example 3 and 4, Table 4 and 5) which also can be found in minor amounts even in the purified product solution (Example 7, Table 9). The chemical impurity has very similar chromatographic properties as F-18 compound and has been identified as 2-{2-[2-(4-{(E)-2-[4-(methylamino)phenyl]vinyl}phenoxy)ethoxy]-ethoxy}ethyl acetate; this compound is formed by the substitution of the mesylate leaving group by acetate present in the eluting solution: This undesired substitution will compete with the [F-18]fluoride substitution affecting both the yield of the desired PET and additionally give more chemical impurities that require removal by HPLC. Surprisingly this acetate substitution and its corresponding impurity can be almost completely avoided if the conventional F-18 eluent mixture is replaced by a solution of kryptofix 2.2.2., potassium carbonate, a low boiling alcohol and water. A. Svadberg et al reported recently on the XII Turku PET Symposium (28-31 May 2011, Turku, Finland) and at the Society of Nuclear Medicine Annual Meeting (4-8 Jun. 2011, San Antonio, USA) about the chemical instability of the conventional eluent mixture. Upon storage of the eluent acetamide and ammonium acetate are formed by alkaline hydrolysis of acetonitrile. The higher the acetate content of the eluent the lower is the radiochemical yield of the F-18 compound and the higher is the content of the formed acetate impurity. So the use of the eluent solution comprising kryptofix 2.2.2, potassium carbonate, a low boiling alcohol and water combines the double advantage giving rise to a cleaner product and a constant good yield which is independent of the storage conditions of the F-18 eluent.
A further major advantage of the new method described herein, is the reliably high radiochemical purity of the F-18 labeled fluoropegylated (aryl/heteroaryl vinyl)-phenyl methyl amines synthesized by the new method. Especially at higher radioactivity levels (>20 GBq) radiochemical purities >95% were achieved.
In a first aspect the present invention is directed to a Method for producing compound of Formula I
comprising the steps of:
X is selected from the group comprising
In one preferred embodiment, X═CH.
In another preferred embodiment, X═N.
R is selected from the group comprising
PG is an “Amine-protecting group”.
In a preferred embodiment, PG is selected from the group comprising:
In a more preferred embodiment, R is H.
In another more preferred embodiment, R is Boc.
LG is a Leaving group.
In a preferred embodiment, LG is selected from the group comprising:
Halogen is chloro, bromo or iodo. Preferably, Halogen is bromo or chloro.
In a preferred embodiment Sulfonyloxy is selected from the group consisting of Methanesulfonyloxy, p-Toluenesulfonyloxy, Trifluormethylsulfonyloxy, 4-Cyanophenylsulfonyloxy, 4-Bromophenylsulfonyloxy, 4-Nitrophenylsulfonyloxy, 2-Nitrophenylsulfonyloxy, 4-Isopropyl-phenylsulfonyloxy, 2,4,6-Triisopropyl-phenylsulfonyloxy, 2,4,6-Trimethylphenylsulfonyloxy, 4-tert-Butyl-phenylsulfonyloxy, 4-Adamantylphenylsulfonyloxy and 4-Methoxyphenylsulfonyloxy.
In a more preferred embodiment, Sulfonyloxy is selected from the group comprising:
b) p-Toluenesulfonyloxy,
In a even more preferred embodiment LG is Methanesulfonyloxy.
In another even more preferred embodiment LG is p-Toluenesulfonyloxy.
A preferred compound of Formula I is:
Another preferred compound of Formula I is:
A preferred compound of Formula II is:
Another preferred compound of Formula II is:
Another preferred compound of Formula II is:
Another preferred compound of Formula II is:
Another preferred compound of Formula II is:
Step 2 comprises a straight forward [F-18]fluoro labeling reaction from compounds of Formula II for obtaining compound of Formula I (if R=H) or compound of Formula III (if R=PG).
The radiolabeling method comprises the step of reacting a compound of Formula II with a F-18 fluorinating agent for obtaining a compound of Formula III or compound of Formula I. In a preferred embodiment, the [F-18]fluoride derivative is 4,7,13,16,21,24-Hexaoxa-1,10-diazabicyclo[8.8.8]-hexacosane K[F-18]F (Kryptofix K[F-18]F), K[F-18]F, H[F-18]F, KH[F-18]F2, Cs[F-18]F, Na[F-18]F or tetraalkylammonium salt of [F-18]F (e.g. [F-18]tetrabutylammonium fluoride). More preferably, the fluorination agent is K[F-18]F, H[F-18]F, [F-18]tetrabutylammonium fluoride, Cs[F-18]F or KH[F-18]F2, most preferably K[F-18], Cs[F-18]F or [F-18]tetrabutylammonium fluoride.
An even more preferred F-18 fluorinating agent is kryptofix/potassium[F-18]fluoride, preferably generated from [F-18]fluoride, kryptofix and potassium carbonate.
The radiofluorination reactions are carried out in acetonitrile, dimethylsulfoxide or dimethylformamide or a mixture thereof. But also other solvents can be used which are well known to someone skilled in the art. Water and/or alcohols can be involved in such a reaction as co-solvent. The radiofluorination reactions are conducted for less than 60 minutes. Preferred reaction times are less than 30 minutes. Further preferred reaction times are less than 15 min. This and other conditions for such radiofluorination are known to experts (Coenen, Fluorine-18 Labeling Methods: Features and Possibilities of Basic Reactions, (2006), in: Schubiger P. A., Friebe M., Lehmann L., (eds), PET-Chemistry—The Driving Force in Molecular Imaging. Springer, Berlin Heidelberg, pp. 15-50).
In one embodiment, 7.5-75 μmol, preferably 10-50 μmol, more preferably 10-30 μmol and even more preferably 12-25 μmol and even more preferably 13-25 μmol of compound of Formula II are used in Step 2.
In another embodiment, more than 7.5 μmol, preferably more than 10 μmol, and more preferable more than 12 μmol and even more preferably more than 13 μmol of compound of Formula II are used in Step 2.
In another embodiment, more than 5 mg, preferably more than 6 mg and more preferably more than 7 mg of compound of Formula II are used in Step 2.
In another embodiment 7 mg of compound of Formula II are used in Step 2.
In another embodiment 8 mg of compound of Formula II are used in Step 2.
In one preferred embodiment, the Radiofluorination of compound of Formula II is carried out in acetonitrile or in a mixture of acetonitrile and co-solvents, wherein the percentage of acetonitrile is at least 50%, more preferably at least 70%, even more preferably at least 90%.
Optionally, if R=PG, Step 3 comprises the deprotection of compound of Formula III to obtain compound of Formula I. Reaction conditions are known or obvious to someone skilled in the art, which are chosen from but not limited to those described in the textbook Greene and Wuts, Protecting groups in Organic Synthesis, third edition, page 494-653, included herewith by reference. Preferred reaction conditions are addition of an acid and stirring at 0° C.-180° C.; addition of an base and heating at 0° C.-180° C.; or a combination thereof.
Preferably the step 2 and step 3 are performed in the same reaction vessel.
Step 4 comprises the purification and optionally formulation of compound of Formula I using a HPLC separation system. Typically, the HPLC solvent eluents are acetonitrile/buffer mixtures. Preferably, a HPLC solvent eluent (e.g. mixtures of ethanol and aqueous buffers) is used, that can be part of the injectable Formulation of compound of Formula I. The collected product fraction can be diluted or mixed with other parts of the Formulation.
In a preferred embodiment, the HPLC solvent mixture is consisting of ethanol or an aqueous buffer or an ethanol/aqueous buffer mixture, wherein the aqueous buffer is consisting of components or excipient that can be injected into human. Examples for such aqueous buffer are solutions of sodium chloride, sodium phosphate buffer, ascorbic acid, ascorbate buffer or mixtures thereof.
In a preferred embodiment, the Method for manufacturing of compound of Formula I is carried out by use of a module (review: Krasikowa, Synthesis Modules and Automation in F-18 labeling (2006), in: Schubiger P. A., Friebe M., Lehmann L., (eds), PET-Chemistry—The Driving Force in Molecular Imaging. Springer, Berlin Heidelberg, pp. 289-316) which allows an automated synthesis. More preferably, the Method is carried out by use of an one-pot module. Even more preferable, the Method is carried out on commonly known non-cassette type modules (e.g. Ecker&Ziegler Modular-Lab, GE Tracerlab FX, Raytest SynChrom) and cassette type modules (e.g. GE Tracerlab MX, GE Fastlab, IBA Synthera, Eckert&Ziegler Modular-Lab PharmTracer), optionally, further equipment such as HPLC or dispensing devices are attached to the said modules.
In a second aspect the present invention is directed to a fully automated and/or remote controlled Method for production of compound of Formula I wherein compounds of Formula I, II and III and Steps 1, 2, 3 and 4 are described above. In a preferred embodiment this method is a fully automated process, compliant with GMP guidelines, that provides a Formulation of Formula I for the use of administration (injection) into human.
In a third aspect the present invention is directed to a Kit for the production of a pharmaceutical composition of compound of Formula I.
The Kit comprises a sealed vial containing kryptofix 2.2.2, potassium carbonate, an alcohol having a boiling point below 100° C. and water.
In one embodiment the Kit comprises a sealed vial containing a predetermined quantity of the compound of Formula II. Preferably, the Kit contains 1.5-75 μmol, preferably 7.5-50 μmol, more preferably 10-50 μmol and even more preferably 12-25 μmol and even more preferably 12-25 μmol and even more preferably 13-25 μmol of compound of Formula II.
In another embodiment the Kit contains more than 7.5 μmol, preferably more than 10 μmol and more preferably more than 12 μmol and even more preferably more than 13 μmol of compound of Formula II.
In another embodiment the Kit contains more than 5 mg, preferably more than 6 mg and more preferably more than 7 mg of compound of Formula II.
In another embodiment the Kit contains 7 mg of compound of Formula II.
In another embodiment the Kit contains 8 mg of compound of Formula II.
The kit also may contain a solvent or solvent mixture or the components for the solvent(mixture) for HPLC purification, such as acetonitrile/buffer mixtures or preferably solvents, solvent mixtures or components that are appropriate for the direct use for injection into patient.
Optionally, the Kit contains further components for manufacturing of compound of Formula I, such as solid-phase extraction cartridges, reagent for fluorination (as described above), acetonitrile or acetonitrile and a co-solvent, reagent for cleavage of deprotection group, solvent or solvent mixtures for purification, solvents and excipient for Formulation.
In one embodiment, the Kit contains a platform (e.g. cassette) for a “cassette-type module” (such as Tracerlab MX or IBA Synthera).
In the context of the present invention, preferred salts are pharmaceutically suitable salts of the compounds according to the invention. The invention also comprises salts which for their part are not suitable for pharmaceutical applications, but which can be used, for example, for isolating or purifying the compounds according to the invention.
Pharmaceutically suitable salts of the compounds according to the invention include acid addition salts of mineral acids, carboxylic acids and sulphonic acids, for example salts of hydrochloric acid, hydrobromic acid, sulphuric acid, phosphoric acid, methanesulphonic acid, ethanesulphonic acid, toluenesulphonic acid, benzenesulphonic acid, naphthalene disulphonic acid, acetic acid, trifluoroacetic acid, propionic acid, lactic acid, tartaric acid, malic acid, citric acid, fumaric acid, maleic acid and benzoic acid.
Pharmaceutically suitable salts of the compounds according to the invention also include salts of customary bases, such as, by way of example and by way of preference, alkali metal salts (for example sodium salts and potassium salts), alkaline earth metal salts (for example calcium salts and magnesium salts) and ammonium salts, derived from ammonia or organic amines having 1 to 16 carbon atoms, such as, by way of example and by way of preference, ethylamine, diethylamine, triethylamine, ethyldiisopropylamine, monoethanolamine, diethanolamine, triethanolamine, dicyclohexylamine, dimethylaminoethanol, procaine, diben-zylamine, N methylmorpholine, arginine, lysine, ethylenediamine and N methylpiperidine.
The term Halogen or halo refers to Cl, Br, F or I.
The term “Amine-protecting group” as employed herein by itself or as part of another group is known or obvious to someone skilled in the art, which is chosen from but not limited to a class of protecting groups namely carbamates, amides, imides, N-alkyl amines, N-aryl amines, imines, enamines, boranes, N—P protecting groups, N-sulfenyl, N-sulfonyl and N-silyl, and which is chosen from but not limited to those described in the textbook Greene and Wuts, Protecting groups in Organic Synthesis, third edition, page 494-653, included herewith by reference. The amine-protecting group is preferably Carbobenzyloxy (Cbz), p-Methoxybenzyl carbonyl (Moz or MeOZ), tert-Butyloxycarbonyl (BOC), 9-Fluorenylmethyloxycarbonyl (FMOC), Benzyl (Bn), p-Methoxybenzyl (PMB), 3,4-Dimethoxybenzyl (DMPM), p-methoxyphenyl (PMP) or the protected amino group is a 1,3-dioxo-1,3-dihydro-2H-isoindol-2-yl (phthalimido) or an azido group.
The term “Leaving group” as employed herein by itself or as part of another group is known or obvious to someone skilled in the art, and means that an atom or group of atoms is detachable from a chemical substance by a nucleophilic agent. Examples are given e.g. in Synthesis (1982), p. 85-125, table 2 (p. 86; (the last entry of this table 2 needs to be corrected: “n-C4F9S(O)2—O— nonaflat” instead of “n-C4H9S(O)2—O— nonaflat”), Carey and Sundberg, Organische Synthese, (1995), page 279-281, table 5.8; or Netscher, Recent Res. Dev. Org. Chem., 2003, 7, 71-83, scheme 1, 2, 10 and 15 and others). (Coenen, Fluorine-18 Labeling Methods: Features and Possibilities of Basic Reactions, (2006), in: Schubiger P. A., Friebe M., Lehmann L., (eds), PET-Chemistry—The Driving Force in Molecular Imaging. Springer, Berlin Heidelberg, pp. 15-50, explicitly: scheme 4 pp. 25, scheme 5 pp 28, table 4 pp 30,
The term Sulfonyloxy refers to
—O—S(O)2-Q wherein Q is optionally substituted aryl or optionally substituted alkyl.
The term “alkyl” as employed herein by itself or as part of another group refers to a C1-C10 straight chain or branched alkyl group such as, for example methyl, ethyl, propyl, isopropyl, butyl, isobutyl, tert-butyl, pentyl, isopentyl, neopentyl, heptyl, hexyl, decyl or adamantyl. Preferably, alkyl is C1-C6 straight chain or branched alkyl or C7-C10 straight chain or branched alkyl. Lower alkyl is a C1-C6 straight chain or branched alkyl.
The term “aryl” as employed herein by itself or as part of another group refers to monocyclic or bicyclic aromatic groups containing from 6 to 10 carbons in the ring portion, such as phenyl, naphthyl or tetrahydronaphthyl.
Whenever the term “substituted” is used, it is meant to indicate that one or more hydrogens on the atom indicated in the expression using “substituted” is/are replaced by one ore multiple moieties from the group comprising halogen, nitro, cyano, trifluoromethyl, alkyl and O-alkyl, provided that the regular valency of the respective atom is not exceeded, and that the substitution results in a chemically stable compound, i.e. a compound that is sufficiently robust to survive isolation to a useful degree of purity from a reaction mixture.
Unless otherwise specified, when referring to the compounds of Formula the present invention per se as well as to any pharmaceutical composition thereof the present invention includes all of the hydrates, salts, and complexes.
The term “F-18” means fluorine isotope 18F. The term“F-19” means fluorine isotope 19F.
Radiochemical and chemical purities of 4-[(E)-2-(4-{2-[2-(2-[F-18]fluoroethoxy)ethoxy]-ethoxy}phenyl)vinyl]-N-methylaniline and 4-[(E)-2-(4-{2-[2-(2-[F-18]fluoroethoxy)ethoxy]-ethoxy}phenyl)vinyl]-N-methylaniline were determined by analytical HPLC (column: Atlantis T3; 150×4.6 mm, 3 μm, Waters; solvent A: 5 mM K2HPO4 pH 2.2; solvent B: acetonitrile; flow: 2 mL/min, gradient: 0:00 min 40% B, 0:00-05:50 min 40-90% B, 05:50-05:60 min 90-40% B, 05:60-09:00 min 40% B).
The synthesis is performed on GE TracerLab MX synthesizer equipped with a customized cassette (
The crude product mixture is diluted with 1.2 mL 2M NaOH and 0.8 mL ammonium formate (1M) from syringe “R5”. 1 mL acetonitrile (from “R2”) and 0.5 mL ethanol (from “R6”) are added separately to the mixture and then transferred to the right syringe of the GE TracerLab MX automate. The reaction mixture containing the crude product is transferred into a 10 ml vial.
The preparation was performed according to the General Procedure for the Synthesis. The composition of the F-18 eluent mixture and the results are listed in Table 2.
The preparation was performed according to the General Procedure for the Synthesis. The composition of the F-18 eluent mixture and the results are listed in Table 3.
Radiosynthesis on Tracerlab MX Using the Conventional QMA Eluent Mixture after Storage of 1 Year at Room Temperature
The preparation was performed according to the General Procedure for the Synthesis. The composition of the F-18 eluent mixture and the results are listed in Table 4.
Radiosynthesis on Tracerlab MX Using the Conventional QMA Eluent Mixture after Storage of 8 Days at 60° C.
The preparation was performed according to the General Procedure for the Synthesis. The composition of the F-18 eluent mixture and the results are listed in the Table 5.
The synthesis is performed on a GE TracerLab MX synthesizer equipped with a customized cassette (
The crude product mixture is diluted with 1.2 mL 2M NaOH and 0.8 mL ammonium formate (1M) from syringe “R5”. 1 mL acetonitrile (from “R2”) and 0.5 mL ethanol (from “R6”) are added separately to the mixture and then transferred to the right syringe of the GE TracerLab MX automate.
The mixture is transferred to the 10 mL sample injection loop of the semi-preparative HPLC using the right syringe of the GE TracerLab MX automate via a liquid sensor which controlls the end of the loading. The mixture is loaded to the semi-preparative HPLC column (Synergi Hydro-RP, 250×10 mm, Phenomenex). A mixture of 60% ethanol and 40% ascorbate buffer (4.32 g/l sodium ascorbate and 0.66 g/l ascorbic acid) is flushed through the column with 6 mL/min. The product fraction is collected directly during 60 sec into the product vial Formulation basis (consisting of 4 ml PEG400 and 16 ml water).
The preparation was performed according to the General Procedure for the Synthesis and purification. The composition of the F-18 eluent mixture and the results are listed in Table 7.
Radiosynthesis on Tracerlab MX Using 2-Propanol as Organic Solvent in the QMA eluent mixture and Eckert&Ziegler Purification Unit
The preparation was performed according to the General Procedure for the Synthesis and purification. The composition of the F-18 eluent mixture and the results are listed in the Table 8.
Radiosynthesis on Tracerlab MX Using the Conventional QMA Eluent Mixture after Storage of 1 Year at Room Temperature and Eckert&Ziegler Purification Unit
The preparation was performed according to the General Procedure for the Synthesis and purification. The composition of the F-18 eluent mixture and the results are listed in Table 9.
| Number | Date | Country | Kind |
|---|---|---|---|
| 11075236.7 | Oct 2011 | EP | regional |
| Filing Document | Filing Date | Country | Kind | 371c Date |
|---|---|---|---|---|
| PCT/EP2012/070669 | 10/18/2012 | WO | 00 | 4/15/2014 |
