U.S. patent application number 13/702007 was filed with the patent office on 2013-08-15 for method for production of f-18 labeled amyloid beta ligands.
This patent application is currently assigned to PIRAMAL IMAGING SA. The applicant listed for this patent is Mathias Berndt, Rainer Braun, Matthias Friebe, Gunnar Garke, Marianne PATT, Fabrice Samson, Andreas Schildan, Christoph Smuda. Invention is credited to Mathias Berndt, Rainer Braun, Matthias Friebe, Gunnar Garke, Marianne PATT, Fabrice Samson, Andreas Schildan, Christoph Smuda.
Application Number | 20130209363 13/702007 |
Document ID | / |
Family ID | 44584930 |
Filed Date | 2013-08-15 |
United States Patent
Application |
20130209363 |
Kind Code |
A1 |
Berndt; Mathias ; et
al. |
August 15, 2013 |
METHOD FOR PRODUCTION OF F-18 LABELED AMYLOID BETA LIGANDS
Abstract
This invention relates to methods, which provide access to
[F-18]fluoropegylated (aryl/heteroaryl vinyl)-phenyl methyl amine
derivatives.
Inventors: |
Berndt; Mathias; (Berlin,
DE) ; Friebe; Matthias; (Berlin, DE) ; Samson;
Fabrice; (Yongsan-gu, KR) ; Braun; Rainer;
(Berlin, DE) ; Garke; Gunnar; (Haan, DE) ;
PATT; Marianne; (Leipzig, DE) ; Schildan;
Andreas; (Leipzig, DE) ; Smuda; Christoph;
(Schlieren, CH) |
|
Applicant: |
Name |
City |
State |
Country |
Type |
Berndt; Mathias
Friebe; Matthias
Samson; Fabrice
Braun; Rainer
Garke; Gunnar
PATT; Marianne
Schildan; Andreas
Smuda; Christoph |
Berlin
Berlin
Yongsan-gu
Berlin
Haan
Leipzig
Leipzig
Schlieren |
|
DE
DE
KR
DE
DE
DE
DE
CH |
|
|
Assignee: |
PIRAMAL IMAGING SA
Matran
CH
|
Family ID: |
44584930 |
Appl. No.: |
13/702007 |
Filed: |
May 30, 2011 |
PCT Filed: |
May 30, 2011 |
PCT NO: |
PCT/EP11/58820 |
371 Date: |
May 2, 2013 |
Current U.S.
Class: |
424/1.89 |
Current CPC
Class: |
A61K 51/121 20130101;
C07B 2200/05 20130101; A61K 51/0455 20130101; A61K 51/04 20130101;
C07B 59/00 20130101; C07C 213/08 20130101; C07C 213/08 20130101;
C07C 217/80 20130101 |
Class at
Publication: |
424/1.89 |
International
Class: |
A61K 51/04 20060101
A61K051/04 |
Foreign Application Data
Date |
Code |
Application Number |
Jun 4, 2010 |
EP |
10164949.9 |
Claims
1. A Method for producing a compound of Formula I ##STR00017##
comprising the steps of: Step 1: Radiolabeling compound of Formula
II with a F-18 fluorinating agent, to obtain compound of Formula I,
if R.dbd.H or to obtain compound of Formula III, if R.dbd.PG
##STR00018## Step 2: If R.dbd.PG, cleavage of the protecting group
PG to obtain a compound of Formula I Step 3: Purification and
Formulation of a compound of Formula I wherein: n=1-6, X is
selected from the group consisting of a) CH, b) N, R is selected
from the group consisting of a) H, b) PG, PG is an
"Amine-protecting group", LG is a leaving group, wherein in step 3
a HPLC method is used, wherein the HPLC solvent or solvent mixture
is part of an injectable Formulation of compound I suitable for
injection into humans.
2. A method according to claim 1, wherein PG is selected from the
group consisting of: a) Boc, b) Trityl and c) 4-Methoxytrityl.
3. A method according to claim 1, wherein LG is selected from the
group consisting of: a) Halogen and b) Sulfonyloxy, Halogen is
chloro, bromo or iodo.
4. A method according to claim 3, wherein Sulfonyloxy is selected
from the group comprising: a) Methanesulfonyloxy, b)
p-Toluenesulfonyloxy, c) (4-Nitrophenyl)sulfonyloxy, d)
(4-Bromophenyl)sulfonyloxy.
5. A method according to claim 1, wherein n=3 and X.dbd.CH.
6. A method according to claim 1, wherein n=3, X.dbd.CH, R=Boc, and
LG=Methanesulfonyloxy.
7. A method according to claim 1, wherein the HPLC solvent is
selected from the group consisting of ethanol, an aqueous buffer or
an ethanol/aqueous buffer mixture.
8. A method according to claim 7, wherein the aqueous buffer is
selected from the group of solutions of sodium chloride, sodium
phosphate buffer, ascorbic acid, ascorbate buffer, or mixtures
thereof.
9. A method according to claim 1, wherein 10-50 .mu.mol of a
compound of Formula II is used.
10. A method according to claim 1, wherein the method is performed
as a fully automated process.
11. A kit, comprising a sealed vial containing a compound of
Formula II and a sealed vial containing a solution for HPLC that
can be part of an injectable Formulation of compound I suitable for
injection into humans.
12. A kit according to claim 10, wherein the solution for HPLC that
can be part of an injectable Formulation of compound I suitable for
injection into humans is selected from the group consisting of
ethanol, an aqueous buffer or an ethanol/aqueous buffer
mixture.
13. A kit according to claim 12, wherein the aqueous buffer is
selected from the group consisting of sodium chloride, sodium
phosphate buffer, ascorbic acid, ascorbate buffer, or mixtures
thereof.
Description
FIELD OF INVENTION
[0001] This invention relates to methods, which provide access to
[F-18]fluoropegylated (aryl/heteroaryl vinyl)-phenyl methyl amine
derivatives.
BACKGROUND
[0002] 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
(A.beta.) and neurofibrillary tangles comprised of paired helical
filaments of hyperphosphorylated tau. The 39-43 amino acids
comprising A.beta. peptides are derived from the larger amyloid
precursor protein (APP). In the amyloidogenic pathway, A.beta.
peptides are cleaved from APP by the sequential proteolysis by
beta- and gamma-secretases. A.beta. 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
A.beta. 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).
[0003] 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.
[0004] 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.
[0005] 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
.beta.+energy (634 keV) is also advantageous.
[0006] 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).
[0007] 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.
[0008] Fluoropegylated (aryl/heteroaryl vinyl)-phenyl methyl amines
such as
4-[(E)-2-(4-{2-[2-(2-fluoroethoxy)ethoxy]ethoxy}phenyl)vinyl]-N-methyl-
aniline and
4-[(E)-2-(6-{2-[2-(2-fluoroethoxy)ethoxy]ethoxy}pyridin-3-yl)vinyl]-N-met-
hylaniline have been labeled with F-18 fluoride and are covered by
patent applications WO2006066104, WO2007126733 and members of the
corresponding patent families.
##STR00001##
[0009] The usefulness of this radiotracers for the detection of
A.beta. 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).
[0010] 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.
[0011] Syntheses of F-18 labeled fluoropegylated (aryl/heteroaryl
vinyl)-phenyl methyl amines have been described before:
4-[(E)-2-(4-{2-[2-(2-[F-18]fluoroethoxy)ethoxy]ethoxy}phenyl)vinyl]-N-meth-
ylaniline
##STR00002##
[0013] a) W. Zhang et al., Nuclear Medicine and Biology 32 (2005)
799-809 [0014] 4 mg precursor 2a
(2-[2-(2-{4-[(E)-2-{4-[(tert-butoxycarbonyl)(methyl)amino]-phenyl}vinyl]p-
henoxy}ethoxy)ethoxy]ethyl methanesulfonate) in 0.2 mL DMSO were
reacted with [F-18]fluoride/kryptofix/potassium carbonate complex.
The intermediate was deprotected with HCl and neutralized with
NaOH. The mixture was extracted with ethyl acetate. The solvent was
dried and evaporated. The residue was dissolved in acetonitrile and
purified by semi-preparative HPLC (acetonitrile/5 mM
dimethylglutarate buffer pH 7 9/1). 20% (decay corrected), 11% (not
corrected for decay)
4-[(E)-2-(4-{2-[2-(2-[F-18]fluoroethoxy)ethoxy]ethoxy}phenyl)vinyl]-N-met-
hylaniline were obtained within 90 min. An additional
re-Formulation, necessary to obtain a solution suitable for
injection into human is not described.
[0015] b) WO2006066104 [0016] 4 mg precursor 2a
(2-[2-(2-{4-[(E)-2-{4-[(tert-butoxycarbonyl)(methyl)amino]-phenyl}vinyl]p-
henoxy}ethoxy)ethoxy]ethyl methanesulfonate) in 0.2 mL DMSO were
reacted with [F-18]fluoride/kryptofix/potassium carbonate complex.
The intermediate was deprotected with HCl and neutralized with
NaOH. The mixture was extracted with ethyl acetate. The solvent was
dried and evaporated, the residue was dissolved in acetonitrile and
purified by semi-preparative HPLC. 30% (decay corrected), 17% (not
corrected for decay)
4-[(E)-2-(4-{2-[2-(2-[F-18]fluoroethoxy)ethoxy]ethoxy}phenyl)vinyl-
]-N-methylaniline were obtained in 90 min. An additional
re-Formulation, necessary to obtain a solution suitable for
injection into human is not described.
[0017] c) C. C. Rowe et al., Lancet Neurology 7 (2008) 129-135
[0018] After radiolabeling, acidic hydrolysis and purification by
semi-preparative H PLC,
4-[(E)-2-(4-{2-[2-(2-[F-18]fluoroethoxy)ethoxy]ethoxy}phenyl)vinyl]-N-met-
hylaniline was Formulated via solid-phase extraction (SPE).
[0019] d) H. Wang et al., Nuclear Medicine and Biology 38 (2011)
121-127
[0020] 5 mg (9.33 .mu.mol) precursor 2a
(2-[2-(2-{4-[(E)-2-{4-[(tert-butoxycarbonyl)(methyl)amino]-phenyl}vinyl]p-
henoxy}ethoxy)ethoxy]ethyl methanesulfonate) in 0.5 mL DMSO were
reacted with [F-18]fluoride/kryptofix/potassium carbonate complex.
The intermediate was deprotected with HCl and neutralized with
NaOH. The crude product was diluted with acetonitrile/0.1 M
ammonium formate (6/4) and purified by semi-preparative HPLC. The
product fraction was collected, diluted with water, passed through
a C18 cartridge and eluted with ethanol, yielding 17% (not
corrected for decay)
4-[(E)-2-(4-{2-[2-(2-[F-18]fluoroethoxy)ethoxy]ethoxy}phenyl)vinyl]-N-met-
hylaniline within 50 min. In the same paper, the conversion of an
unprotected mesylate precursor (is described: [0021] 5 mg (11.48
.mu.mol) unprotected mesylate precursor
(2-{2-[2-(4-{(E)-2-[4-(methylamino)phenyl]vinyl}phenoxy)ethoxy]-ethoxy}et-
hyl 4-methanesulfonate) in 0.5 mL DMSO were reacted with
[F-18]fluoride/kryptofix/potassium carbonate complex. The crude
product was diluted with acetonitrile/0.1 M ammonium formate (6/4)
and purified by semi-preparative HPLC. The product fraction was
collected, diluted with water, passed through a C18 cartridge and
eluted with ethanol, yielding 23% (not corrected for decay)
4-[(E)-2-(4-{2-[2-(2-[F-18]fluoroethoxy)ethoxy]ethoxy}phenyl)vinyl]-N-met-
hylaniline within 30 min. [0022] A process wherein the radiotracer
was purified by SPE (without HPLC) only, was found to afford a
product with acceptable radiochemical purity (>95%), however,
the chemical purity was too low, e.g. side products derived from
the excess of precursor could not be removed.
[0023] e) US20100113763 [0024] 2a
(2-[2-(2-{4-[(E)-2-{4-[(tert-butoxycarbonyl)(methyl)amino]phenyl}vinyl]-p-
henoxy}ethoxy)ethoxy]ethyl methanesulfonate) was reacted with
[F-18]fluoride reagent in a mixture of tert-alcohol and
acetonitrile. After fluorination, the solvent was evaporated and a
mixture of HCl and acetonitrile was added. After deprotection
(heating at 100-120.degree. C.), the crude product mixture was
purified by HPLC(C18, 60% acetonitrile, 40% 0.1M ammonium formate).
An additional re-Formulation, necessary to obtain a solution
suitable for injection into human is not described.
4-[(E)-2-(6-{2-[2-(2-[F-18]fluoroethoxy)ethoxy]ethoxy}pyridin-3-yl)vinyl]--
N-methylaniline
##STR00003##
[0026] a) S. R. Choi et al., The Journal of Nuclear Medicine 50
(2009) 1887-1894. [0027] 1 mg precursor 2b
(2-{2-[2-({5-[(E)-2-{4-[(tert-butoxycarbonyl)(methyl)amino]-phenyl}vinyl]-
pyridin-2-yl}oxy)ethoxy]ethoxy}ethyl 4-methylbenzenesulfonate) in 1
mL DMSO was reacted with [F-18]fluoride/kryptofix/potassium
carbonate complex. The intermediate was deprotected with HCl and
neutralized with NaOH. DMSO and inorganic components were removed
by solid-phase-extraction on SepPak light C18 cartridge (Waters).
The crude product was purified by semi-preparative HPLC (55%
acetonitrile, 45% 20 mM NH.sub.4OAc+0.5% w/v sodium ascorbate). The
product fraction was diluted with water and passed through a SepPak
light C18 cartridge. The radiotracer was eluted with ethanol. The
yield for
4-[(E)-2-(6-{2-[2-(2-[F-18]fluoroethoxy)ethoxy]ethoxy}pyridin-3-yl)vinyl]-
-N-methylaniline was 10-30% (decay corrected).
[0028] b) WO2010078370 [0029] 1.5 mg (2.45 .mu.mol) precursor 2b
(2-{2-[2-({5-[(E)-2-{4-[(tert-butoxycarbonyl)(methyl)amino]-phenyl}vinyl]-
pyridin-2-yl}oxy)ethoxy]ethoxy}ethyl 4-methylbenzenesulfonate) in 2
mL DMSO was reacted with [F-18]fluoride/kryptofix/potassium
carbonate complex. The intermediate was deprotected with HCl and
diluted with 1% NaOH solution for neutralization. The mixture was
loaded onto a reverse phase cartridge. The cartridge was washed
with water (containing 5% w/v sodium ascorbate). The crude product
was eluted with acetonitrile into a reservoir containing water+5%
w/v sodium ascorbate and HPLC solvent. After purification by
semi-preparative HPLC, the product fraction was collected into a
reservoir containing water+0.5% w/v sodium ascorbate. The solution
was passed trough a C18 cartridge, the cartridge was washed with
water (containing 0.5% w/v sodium ascorbate and the final product
was eluted with ethanol into a vial containing 0.9% sodium chloride
solution with 0.5% w/v sodium ascorbate.
[0030] c) Y. Liu et al., Nuclear Medicine and Biology 37 (2010)
917-925 [0031] 1 mg (1.63 .mu.mol) precursor 2b
(2-{2-[2-({5-[(E)-2-{4-[(tert-butoxycarbonyl)(methyl)amino]-phenyl}vinyl]-
pyridin-2-yl}oxy)ethoxy]ethoxy}ethyl 4-methylbenzenesulfonate) in 1
mL DMSO was reacted with [F-18]fluoride/kryptofix/potassium
carbonate complex. The intermediate was deprotected with HCl and
diluted with 1% NaOH solution. The mixture was loaded onto a Oasis
HLB cartridge. The cartridge was washed with water, dried under a
flow of argon and the product was eluted with ethanol into a vial
containing a saline solution. Although, radiochemical impurities
were removed by this procedure, non-radioactive by-products derived
from hydrolysis of the excess of precursor, remained in the final
product solution. [0032] The yield for
4-[(E)-2-(6-{2-[2-(2-[F-18]fluoroethoxy)ethoxy]ethoxy}pyridin-3-yl)vinyl]-
-N-methylaniline was 34% (non-decay corrected) within 50 min at a
radioactive level from 10-100 mCi (370-3700 MBq) of
[F-18]fluoride.
[0033] d) L. Silva et al., Abstract/Poster EANM 2010 [0034] An IBA
Synthera platform was adapted for the synthesis of
4-[(E)-2-(6-{2-[2-(2-[F-18]fluoroethoxy)ethoxy]ethoxy}pyridin-3-yl)vinyl]-
-N-methylaniline. Additionally, a semi-preparative HPLC system and
a further Synthera module for re-Formulation was integrated.
[0035] e) G. Casale et al. World Journal of Nuclear Medicine, 9 S1
(2010), S-174 (Abstract of 10.sup.th Congress of WFNMB, Cape Town,
South Africa, 18-23 Sep. 2010) [0036] The manufacturing of
4-[(E)-2-(6-{2-[2-(2-[F-18]fluoroethoxy)ethoxy]ethoxy}pyridin-3-yl)vinyl]-
-N-methylaniline have been accomplished by use of an IBA Synthera
synthesis module, combined with an HPLC semi preparative
purification system and an additional module for Formulation
(dilution of HPLC fraction, trapping on a C18 cartridge, washing
and elution with ethanol).
[0037] 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. So far, F-18
labeled fluoropegylated (aryl/heteroaryl vinyl)-phenyl methyl
amines have been purified by HPLC using solvent systems consisting
of acetonitrile and aqueous buffer. Obviously, collected product
fractions can not directly used for administration into patient.
Acetonitrile and further compounds of the solvent systems that are
not tolerated for injection into human have to be removed. This
could be accomplished by evaporation or by solid phase extraction
(e.g. trapping on C18 solid phase extracting cartridge and elution
with ethanol, see FIG. 1: final solid-phase extraction cartridge
C3, elution with ethanol from V8; see also FIG. 7, final
solid-phase extraction cartridge 11, elution with ethanol from one
of the vials 9).
[0038] However, especially at higher levels of radioactivity,
decomposition of the radiotracer due to radiolysis processes might
be an issue. This problem is well known, to prevent radiolysis
during the purification of
4-[(E)-2-(6-{2-[2-(2-[F-18]fluoroethoxy)ethoxy]ethoxy}pyridin-3-yl)vinyl]-
-N-methylaniline sodium ascorbate (as an radical scavenger) was
added to the HPLC solvent and to washing solutions (S. R. Choi et
al, WO2010078370). However, the concentration of the radiotracer
after HPLC by evaporation or by solid-phase extraction is a
critical step of the manufacturing. In upscaling experiments,
higher radiochemical purities of F-18 labeled fluoropegylated
(aryl/heteroaryl vinyl)-phenyl methyl amines can be found after
HPLC, before the solid phase extraction compared to the composition
after solid phase extraction.
[0039] The general setup of the manufacturing process for F-18
labeled fluoropegylated (aryl/heteroaryl vinyl)-phenyl methyl
amines as previously described is illustrated in FIG. 7. The
manufacturing process can be divided into three major parts: [0040]
A) Synthesis [0041] B) Purification by HPLC [0042] C)
Formulation
[0043] The manufacturing steps of drying of [F-18]fluoride,
radiolabeling of the precursor molecule and deprotection are
performed on the part A of the synthesis device (FIG. 7). The crude
product mixture is transferred to the second part B for
purification by HPLC (on reversed phase silica gel using
acetonitrile/buffer eluent). To obtain the radiotracer in a
Formulation, suitable for injection into human. The solvent
(acetonitrile) present in the product fraction needs to be removed
and exchanged by a composition that is appropriate for the
manufacturing of a medicament. Typically (and described in the
references above), the product fraction is diluted with water
(vessel "8", FIG. 7, part C) and then passed through a reversed
phase cartridge ("11", FIG. 7, part C). The cartridge is washed
with a aqueous solution from one of the reservoirs 9 (FIG. 7, part
C) and finally eluted from the cartridge with an ethanolic solution
(or ethanol) from another of the reservoirs 9 into the product
vial, that optionally comprises further parts and excipients of the
final Formulation. It is obvious to those skilled in the art, that
the illustration in FIG. 7 is a simplification of process and
equipment and that further parts such as valves, vials, tubing ect.
can be part of such process or equipment.
[0044] 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%.
[0045] Although ascorbate/ascorbic acid is added to solvents
involved in the purification, radiochemical purity was only about
95.3.+-.2.2% at product activity levels of up to 18.5 GBq (Yao et
al.)--probably due to decomposition by radiolysis. At higher
product activity levels an even higher variation of radiochemical
purity was found for the manufacturing of
4-[(E)-2-(4-{2-[2-(2-[F-18]fluoroethoxy)ethoxy]-ethoxy}phenyl)vinyl]-N-
-methylaniline (Example 7, FIG. 9, method A).
[0046] Beside of the variation of radiochemical purity, the
re-Formulation during the current process (conversion of the
radiotracer from HPLC media into an injectable solution) requires
additional process time and demands more complex equipment. For
example, the process for the synthesis of
4-[(E)-2-(6-{2-[2-(2-[F-18]fluoroethoxy)ethoxy]ethoxy}pyridin-3-yl)vinyl]-
-N-methylaniline described by Silva et al. and Casale et al.
demands three modules for the overall manufacturing procedure. The
Synthesis of the crude product (schematically illustrated in FIG.
7, Part A) was accomplished on an IBA Synthera module, a
semi-preparative HPLC system was used for purification
(schematically illustrated in FIG. 7, Part B) and an additional IBA
Synthera synthesis module was used for re-Formulation
(schematically illustrated in FIG. 7, Part C).
[0047] The problem to be solved by the present invention is to
provide an improved HPLC purification process for F-18 labeled
fluoropegylated (aryl/heteroaryl vinyl)-phenyl methyl amines that
provides high chemical and radiochemical purities of the
radiotracer, avoiding a concentration of the labeled product after
purification to prevent radiolysis, especially at higher levels of
radioactivity. Such process should be suitable for the
manufacturing of larger quantities (radioactivity) of the
radiotracer to allow a distribution to imaging facilities without
own radiopharmaceutical production. So far the maximum activity for
a F-18 labeled fluoropegylated (aryl/heteroaryl vinyl)-phenyl
methyl amine was reported to be 18.5 GBq (Yao et al.). However,
even higher yields would be supportive for a widespread use and
availability of the radiotracer. A prerequisite of the new
manufacturing method should be a high radiochemical purity (e.g.
>95%) within a broad range of radioactivity. More precisely,
such process should be suitable for the manufacturing of higher
activity levels of the radiotracer than previously described (e.g.
>20 GBq, or even >50 GBq, or even >100 GBq) with
radiochemical purities reliably .gtoreq.95%. As an additional
feature such process should be less complex than the processes
described before.
[0048] The problems described above were solved by an modified
purification procedure. To simplify the overall setup for
manufacturing, the solvent composition for HPLC purification was
modified. Instead of an acetonitrile/buffer mixture, an
ethanol/buffer mixture is used. An advantage of the new HPLC
solvent mixture is, that all constituents of the HPLC solvent--in
contrast to previously described compositions--are well tolerated
as part of a Formulation, thereby suitable for injection into
human. Therefore a re-Formulation to remove constituents of the
HPLC solvent (as illustrated in FIG. 7, Part C) is not longer
required. This further advantage of the new process--the simplified
setup--is schematically illustrated in FIG. 8. (Obviously, this
illustration is a simplification that shows a general setup of the
new method described herein.) Following the drawing in FIG. 8, the
product fraction is collected directly (by switching valve "7")
into the product vial (that could contain further parts of the
final Formulation). Due to the reduced complexity, the overall
manufacturing time by using the new method described herein is
shorter, directly contributing to higher non decay corrected yields
compared to the previous used process wherein a HPLC purification
with additional (time consuming) re-Formulation on a solid-phase
cartridge (SPE) is used.
[0049] The 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. In Example 7 and FIG. 9 the
radiochemical purity in dependence of purification method and
amount (radioactivity) of radiolabeled product at end of synthesis
is demonstrated. The dots/squares (each representing an individual
experiment) and the trendlines in FIG. 9 clearly demonstrate that
the radiochemical purity obtained after HPLC with re-Formulation by
SPE varies significantly (FIG. 9, empty squares). Especially at
higher radioactivity levels (>20 GBq) the radiochemical purity
often is even .ltoreq.95%. In contrast, variability of
radiochemical purities obtained by the new method of the present
invention is much lower and high radiochemical purities of >95%
were achieved, even at radioactivity levels of the product of
greater than 50 GBq or even greater than 100 GBq (FIG. 9, filled
dots).
SUMMARY OF THE INVENTION
[0050] The present invention provides a Method for production of
radiolabeled compound of Formula I and suitable salts of an
inorganic or organic acid thereof, hydrates, complexes, esters,
amides, solvates and prodrugs thereof and a optionally a
pharmaceutically acceptable carrier, diluent, adjuvant or
excipient. [0051] The method comprises the steps of: [0052]
Radiofluorination of compound of Formula II [0053] Optionally,
cleavage of a protecting group [0054] Purification and Formulation
of compound of Formula I by HPLC using a solvent system that can be
part of an injectable Formulation
[0054] ##STR00004## [0055] The Method provided by the present
invention is schematically illustrated in FIG. 8. Radiofluorination
of compound of Formula II and optionally, the cleavage of a
protecting group are performed on the left-hand part of the
equipment (FIG. 8, part A). The purification of compound of Formula
I is performed in a way, that the product fraction obtained by HPLC
(FIG. 8, part B) can be directly transferred into the product vial,
wherein the product vial optionally contains further
pharmaceutically acceptable carriers, diluents, adjuvant or
excipients. A further part of process and equipment as illustrated
in FIG. 7 (Part C) is not longer required by the Method of the
present invention. [0056] The present invention also provides
compositions comprising a radiolabeled compound of Formula I or
suitable salts of an inorganic or organic acid thereof, hydrates,
complexes, esters, amides, solvates and prodrugs thereof and
optionally a pharmaceutically acceptable carrier, diluent, adjuvant
or excipient. [0057] The present invention also provides a Kit for
preparing a radiopharmaceutical preparation by the herein described
process, said Kit comprising a sealed vial containing a
predetermined quantity of the compound of Formula II.
DESCRIPTION OF THE INVENTION
[0058] In a first aspect the present invention is directed to a
Method for producing compound of Formula I
##STR00005##
comprising the steps of: [0059] Step 1: Radiolabeling compound of
Formula II with a F-18 fluorinating agent, to obtain compound of
Formula I, if R.dbd.H or to obtain compound of Formula III, if
R.dbd.PG
[0059] ##STR00006## [0060] Step 2: Optionally, if R.dbd.PG,
cleavage of the protecting group PG to obtain compound of Formula I
[0061] Step 3: Purification and Formulation of compound of Formula
I wherein:
[0062] n=1-6, preferably 2-4, more preferably 3.
[0063] X is selected from the group comprising
a) CH,
b) N.
[0064] In one preferred embodiment, X.dbd.CH.
[0065] In another preferred embodiment, X.dbd.N.
[0066] R is selected from the group comprising
a) H,
b) PG.
[0067] PG is an "Amine-protecting group".
[0068] In a preferred embodiment, PG is selected from the group
comprising:
a) Boc,
b) Trityl and
c) 4-Methoxytrityl.
[0069] In a more preferred embodiment, R is H.
[0070] In another more preferred embodiment, R is Boc.
[0071] LG is a Leaving group.
[0072] In a preferred embodiment, LG is selected from the group
comprising:
a) Halogen and
b) Sulfonyloxy.
[0073] Halogen is chloro, bromo or iodo. Preferably, Halogen is
bromo or chloro.
[0074] 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.
[0075] In a more preferred embodiment, Sulfonyloxy is selected from
the group comprising:
a) Methanesulfonyloxy,
[0076] b) p-Toluenesulfonyloxy,
c) (4-Nitrophenyl)sulfonyloxy,
d) (4-Bromophenyl)sulfonyloxy.
[0077] In a even more preferred embodiment LG is
Methanesulfonyloxy.
[0078] In another even more preferred embodiment LG is
p-Toluenesulfonyloxy.
[0079] A preferred compound of Formula I is:
##STR00007##
4-[(E)-2-(4-{2-[2-(2-[F-18]fluoroethoxy)ethoxy]ethoxy}phenyl)vinyl]-N-met-
hylaniline
[0080] Another preferred compound of Formula I is:
##STR00008##
4-[(E)-2-(6-{2-[2-(2-[F-18]fluoroethoxy)ethoxy]ethoxy}pyridin-3-yl)vinyl]-
-N-methylaniline
[0081] A preferred compound of Formula II is:
##STR00009##
2-[2-(2-{4-[(E)-2-{4-[(tert-butoxycarbonyl)(methyl)amino]phenyl}vinyl]phe-
noxy}-ethoxy)ethoxy]ethyl methanesulfonate
[0082] Another preferred compound of Formula II is:
##STR00010##
2-[2-(2-{4-[(E)-2-{4-[(tert-butoxycarbonyl)(methyl)amino]phenyl}vinyl]phe-
noxy}-ethoxy)ethoxy]ethyl 4-methylbenzenesulfonate
[0083] Another preferred compound of Formula II is:
##STR00011##
2-{2-[2-(4-{(E)-2-[4-(methylamino)phenyl]vinyl}phenoxy)ethoxy]ethoxy}ethy-
l 4-methylbenzenesulfonate
[0084] Another preferred compound of Formula II is:
##STR00012##
2-{2-[2-(4-{(E)-2-[4-(methylamino)phenyl]vinyl}phenoxy)ethoxy]ethoxy}ethy-
l 4-methylbenzenesulfonate
[0085] Another preferred compound of Formula II is:
##STR00013##
2-{2-[2-({5-[(E)-2-{4-[(tert-butoxycarbonyl)(methyl)amino]phenyl}vinyl]py-
ridin-2-yl}oxy)ethoxy]ethoxy}ethyl 4-methylbenzenesulfonate
[0086] Step 1 comprises a straight forward [F-18]fluoro labeling
reaction from compounds of Formula II for obtaining compound of
Formula I (if R.dbd.H) or compound of Formula III (if
R.dbd.PG).
[0087] 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]F.sub.2,
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]F.sub.2, most preferably K[F-18],
Cs[F-18]F or [F-18]tetrabutylammonium fluoride.
[0088] An even more preferred F-18 fluorinating agent is
kryptofix/potassium[F-18]fluoride, preferably generated from
[F-18]fluoride, kryptofix and potassium carbonate.
[0089] 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).
[0090] In one embodiment, 7.5-75 .mu.mol, preferably 10-50 .mu.mol,
more preferably 10-30 .mu.mol and even more preferably 12-25
.mu.mol and even more preferably 13-25 .mu.mol of compound of
Formula II are used in Step 1.
[0091] In another embodiment, more than 7.5 .mu.mol, preferably
more than 10 .mu.mol, and more preferable more than 12 .mu.mol and
even more preferably more than 13 .mu.mol of compound of Formula II
are used in Step 1.
[0092] 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 1.
[0093] In another embodiment 7 mg of compound of Formula II are
used in Step 1.
[0094] In another embodiment 8 mg of compound of Formula II are
used in Step 1.
[0095] 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%.
[0096] Optionally, if R.dbd.PG, Step 2 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.degree. C.-180.degree. C.; addition of an base
and heating at 0.degree. C.-180.degree. C.; or a combination
thereof.
[0097] Preferably the step 1 and step 2 are performed in the same
reaction vessel.
[0098] Step 3 comprises the purification and Formulation of
compound of Formula I using a HPLC separation system, wherein, the
HPLC solvent eluent (e.g. mixtures of ethanol and aqueous buffers)
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.
[0099] 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.
[0100] Examples for such aqueous buffer are solutions of sodium
chloride, sodium phosphate buffer, ascorbic acid, ascorbate buffer
or mixtures thereof.
[0101] 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.
[0102] 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 and 3 are described above.
[0103] 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.
[0104] In a third aspect the present invention is directed to a Kit
for the production of a pharmaceutical composition of compound of
Formula I.
[0105] In one embodiment the Kit comprising a sealed vial
containing a predetermined quantity of the compound of Formula II.
Preferably, the Kit contains 1.5-75 .mu.mol, preferably 7.5-50
.mu.mol, more preferably 10-50 .mu.mol and even more preferably
12-25 .mu.mol and even more preferably 12-25 .mu.mol and even more
preferably 13-25 .mu.mol of compound of Formula II.
[0106] In another embodiment the Kit contains more than 7.5
.mu.mol, preferably more than 10 .mu.mol and more preferably more
than 12 .mu.mol and even more preferably more than 13 .mu.mol of
compound of Formula II.
[0107] 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.
[0108] In another embodiment the Kit contains 7 mg of compound of
Formula II.
[0109] In another embodiment the Kit contains 8 mg of compound of
Formula II.
[0110] The kit also contains a solvent or solvent mixture or the
components for the solvent (mixture) for HPLC purification, wherein
those solvent, solvent mixture or components are appropriate for
the direct use for injection into patient.
[0111] 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.
[0112] In one embodiment, the Kit contains a platform (e.g.
cassette) for a "cassette-type module" (such as Tracerlab MX or IBA
Synthera).
DEFINITIONS
[0113] 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.
[0114] 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.
[0115] 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.
[0116] The term Halogen or halo refers to Cl, Br, F or I.
[0117] 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.
[0118] 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-C.sub.4F.sub.9S(O).sub.2--O-- nonaflat" instead of
"n-C.sub.4H.sub.9S(O).sub.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, FIG. 7
pp 33).
[0119] The term Sulfonyloxy refers to
--O--S(O).sub.2-Q wherein Q is optionally substituted aryl or
optionally substituted alkyl.
[0120] The term "alkyl" as employed herein by itself or as part of
another group refers to a C.sub.1-C.sub.10 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
C.sub.1-C.sub.6 straight chain or branched alkyl or
C.sub.7-C.sub.10 straight chain or branched alkyl. Lower alkyl is a
C.sub.1-C.sub.6 straight chain or branched alkyl.
[0121] 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.
[0122] 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.
[0123] 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.
[0124] The term "F-18" means fluorine isotope .sup.18F. The
term"F-19" means fluorine isotope .sup.19F.
EXAMPLES
Determination of Radiochemical and Chemical Purity
[0125] Radiochemical and chemical purities of
4-[(E)-2-(4-{2-[2-(2-[F-18]fluoroethoxy)ethoxy]-ethoxy}phenyl)vinyl]-N-me-
thylaniline and
4-[(E)-2-(4-{2-[2-(2-[F-18]fluoroethoxy)ethoxy]-ethoxy}phenyl)vinyl]-N-me-
thylaniline were determined by analytical HPLC (column: Atlantis
T3; 150.times.4.6 mm, 3 .mu.m, Waters; solvent A: 5 mM
K.sub.2HPO.sub.4 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). [0126] Retention time of
4-[(E)-2-(4-{2-[2-(2-[F-18]fluoroethoxy)ethoxy]-ethoxy}phenyl)-vinyl]-N-m-
ethylaniline: 3.5-3.9 min depending on the HPLC system used for
quality control. Due to different equipment (e.g. tubing) a
difference in retention time is observed between the different HPLC
systems. The identity of
4-[(E)-2-(4-{2-[2-(2-[F-18]fluoroethoxy)ethoxy]-ethoxy}phenyl)vinyl]-N-me-
thylaniline was proofed by co-injection with the non-radioactive
reference
4-[(E)-2-(4-{2-[2-(2-[F-19]fluoroethoxy)ethoxy]-ethoxy}phenyl)vinyl]-N-me-
thylaniline. [0127] Retention time of
4-[(E)-2-(6-{2-[2-(2-[F-18]fluoroethoxy)ethoxy]-ethoxy}pyridin-3-yl)vinyl-
]-N-methylaniline: 3.47 min. The identity of
4-[(E)-2-(6-{2-[2-(2-[F-18]fluoroethoxy)ethoxy]-ethoxy}pyridin-3-yl)vinyl-
]-N-methylaniline was proofed by co-elution with the
non-radioactive reference
-[(E)-2-(6-{2-[2-(2-[F-19]fluoroethoxy)ethoxy]-ethoxy}pyridin-3-
-yl)vinyl]-N-methylaniline.
Example 1
Synthesis of
4-[(E)-2-(4-{2-[2-(2-[F-18]fluoroethoxy)ethoxy]-ethoxy}phenyl)vinyl]-N-me-
thylaniline Radiosynthesis on Eckert&Ziegler Modular Lab
##STR00014##
[0129] The synthesis of
4-[(E)-2-(4-{2-[2-(2-[F-18])fluoroethoxy)ethoxy]-ethoxy}phenyl)vinyl]-N-m-
ethylaniline have been performed on a Eckert&Ziegler modular
lab synthesizer. [F-18]Fluoride (60362 MBq) was trapped on a QMA
cartridge. The activity was eluted with potassium
mesylate/kryptofix/n-Bu.sub.4NHCO.sub.3/methanol mixture into the
reactor. The solvent was removed while heating under gentle
nitrogen stream and vacuum. Drying was repeated after addition of
acetonitrile. A solution of 4 mg 2a in 1 mL
tert-amylalcohol/acetonitrile (9:1) was added to the dried residue
and the mixture was heated for 20 min at 120.degree. C. During
heating, the exhaust of the reactor was opened to allow the
evaporation of the solvent. A mixture of 2.2 mL 1.5M HCl, 1.1 mL
acetonitrile and 30 mg sodium ascorbate was added and the reactor
was heated at 100.degree. C. for 10 min. The crude product was
neutralized (1.5 mL 2M NaOH+0.3 mL buffer) and transferred to a
semi-preparative HPLC column (Synergy Hydro-RP, 250.times.10 mm,
Phenomenex). A mixture of 60% ethanol and 40% ascorbate buffer (pH
7.0) was flushed through the column with 3 mL/min. The product
fraction at .apprxeq.18 min (FIG. 2) was directly collected into
the product vial containing 8.5 mL Formulation basis (phosphate
buffer, ascorbic acid, PEG400). Analytical HPLC of the final
product (FIG. 3) showed excellent radiochemical and chemical
purity. Only cold
4-[(E)-2-(4-{2-[2-(2-[F-18]fluoroethoxy)ethoxy]-ethoxy}phenyl)vinyl]-N-me-
thylaniline have been detected in the UV chromatogram (FIG. 3,
bottom), all non-radioactive impurities have been separated. The
radiochemical purity was determined to be 99.6%.
Example 2
Synthesis of
4-[(E)-2-(4-{2-[2-(2-[F-18]fluoroethoxy)ethoxy]-ethoxy}phenyl)vinyl]-N-me-
thylaniline Radiosynthesis on Tracerlab FX.sub.N
[0130] A Tracerlab FX.sub.N synthesizer have been adopted to the
"direct cut HPLC approach" (FIG. 4).
[0131] [F-18]Fluoride (3700 MBq) was trapped on a QMA cartridge.
The activity was eluted with potassium
carbonate/kryptofix/acetonitrile/water mixture into the reactor.
The solvent was removed while heating under gentle nitrogen stream
and vacuum. Drying was repeated after addition of acetonitrile. A
solution of 7 mg 2a in 1 mL acetonitrile was added to the dried
residue and the mixture was heated for 8 min at 120.degree. C.
After cooling to 60.degree. C., a mixture of 0.5 mL 2M HCl, and 0.5
mL acetonitrile was added and the reactor was heated at 110.degree.
C. for 4 min. The crude product was neutralized (1 mL 1M NaOH+2 mL
buffer) and transferred to a semi-preparative HPLC column (Synergy
Hydro-RP, 250.times.10 mm, Phenomenex). A mixture of 60% ethanol
and 40% ascorbate buffer (pH 7.0) was flushed through the column
with 3 mL/min. The product fraction at .apprxeq.16 min (FIG. 2) was
directly collected into the product vial containing 8.5 Formulation
basis (phosphate buffer, ascorbic acid, PEG400). Radiochemical
purity was determined to be >99%.
Example 3
Synthesis of
4-[(E)-2-(4-{2-[2-(2-[F-18]fluoroethoxy)ethoxy]-ethoxy}phenyl)vinyl]-N-me-
thylaniline Radiosynthesis on Tracerlab MX and Eckert&Ziegler
Purification Unit
[0132] A Kit have been assembled for the synthesis of
4-[(E)-2-(4-{2-[2-(2-[F-18]fluoroethoxy)ethoxy]-ethoxy}phenyl)vinyl]-N-me-
thylaniline (Table 1).
TABLE-US-00001 TABLE 1 Composition of Kit for manufacturing of
4-[(E)-2-(4-{2-[2-
(2-[F-18]fluoroethoxy)ethoxy]-ethoxy}phenyl)vinyl]-N- methylaniline
on tracerlab MX and Eckert&Ziegler Purification unit Eluent
vial 22 mg kryptofix. 7 mg potassium carbonate in 300 .mu.L water +
300 .mu.L acetonitrile Blue capped vial 8 mL acetonitrile Red
capped vial 8 mg precursor 2a Green capped vial 2 mL 1.5M HCl + 30
mg sodium ascorbate 2 mL syringe 1.5 mL 2M NaOH + 0.3 mL phosphate
buffer Water bag Water Product line to Tube with two luer lock
fittings Eckert&Ziegler purification unit Anion exchange
cartridge QMA light, Waters (pre-conditioned) Disposable 3-way
valve With tubing and needle to product vial, incl. sterile filters
Product vial 20 mL vial Formulation basis 8.5 mL (PEG 400,
Na.sub.2HPO.sub.4.cndot.H.sub.2O, ascorbic acid in water) HPLC
solvent ethanol water sodium ascorbate ascorbic acid HPLC flow rate
3 mL/min
[0133] The design of the Tracerlab MX cassette has been adopted
(FIG. 5). [F-18]Fluoride was trapped on the QMA cartridge. The
activity was eluted with potassium
carbonate/kryptofix/acetonitrile/water mixture (from "eluent vial")
into the reactor. The solvent was removed while heating under
gentle nitrogen stream and vacuum. Drying was repeated after
addition of acetonitrile. A solution of 8 mg 2a in 1.8 mL
acetonitrile (acetonitrile from "blue capped vial" was added to
solid 2a in the "red capped vial" during the sequence) was added to
the dried residue and the mixture was heated for 10 min at
120.degree. C. 1.5M HCl (from "green capped vial") was added and
the reactor was heated at 110.degree. C. for 5 min. The crude
product was neutralized (1 mL 1M NaOH+0.3 mL buffer, from "2 mL
syringe") and transferred to the injection valve of the
Eckert&Ziegler HPLC (FIG. 6) by the left syringe pump of the MX
module. The crude product was purified on a Synergy Hydro-RP,
250.times.10 mm, Phenomenex HPLC column using a mixture of 60%
ethanol and 40% ascorbate buffer (pH 7.0). The product fraction at
.apprxeq.17.5 min (FIG. 2) was directly collected into the product
vial containing 8.5 Formulation basis (phosphate buffer, ascorbic
acid, PEG400).
Example 4
Synthesis of
4-[(E)-2-(4-{2-[2-(2-[F-18]fluoroethoxy)ethoxy]-ethoxy}phenyl)vinyl]-N-me-
thylaniline Radiosynthesis on Eckert&Ziegler Modular Lab
[0134] The synthesis has been performed on Eckert & Ziegler
ModularLab synthesizer using acetonitrile as solvent for
fluorination. The setup of the synthesizer and the results are
summarized in Table 2.
[0135] [F-18]Fluoride was trapped on a QMA cartridge (C1). The
activity was eluted with a kryptofix mixture (from "V1") into the
reactor. The solvent was removed while heating under gentle
nitrogen stream and vacuum. Drying was repeated after addition of
100 .mu.L acetonitrile (from "V2"). The solution of precursor 2a
(from "V3") was added to the dried residue and the mixture was
heated for 10 min at 120.degree. C. After cooling to 40.degree. C.,
2 mL 1.5M HCl (from "V4") was added and solution was heated for 5
min at 110.degree. C.
[0136] The crude product mixture was diluted with 1.2 mL 2M NaOH
and 0.8 mL ammonium formate (1 M) from vial "V5" and then
transferred to the HPLC vial ("Mix-Vial") containing previously 1
mL acetonitrile and 0.5 mL ethanol. The mixture was transferred to
the 10 mL sample injection loop of the semi-preparative HPLC using
a nitrogen overpressure in the HPLC vial ("Mix-Vial") and via a
liquid sensor which controlled the end of the loading. The mixture
is loaded to the semi-preparative HPLC column (Synergi Hydro-RP,
250.times.10 mm, Phenomenex). A mixture of 60% ethanol and 40%
ascorbate buffer was flushed through the column with 6 mL/min. The
product fraction at .apprxeq.7 min was collected directly into the
product vial containing 15 mL Formulation basis (consisting of
phosphate buffer, PEG400 and ascorbic acid). Analytical HPLC of the
final product showed excellent radiochemical and chemical purity.
No impurity higher than 0.3 .mu.g/mL was quantified.
TABLE-US-00002 TABLE 2 Vial V1 22 mg kryptofix 7 mg potassium
carbonate 300 .mu.L acetonitrile 300 .mu.L water Vial V2 100 .mu.L
acetonitrile Vial V3 8 mg precursor 2a in 1.8 mL acetonitrile Vial
V4 2 mL HCl 1.5M Vial V5 1.2 mL NaOH 2.0M 800 .mu.L ammonium
formate 1M Cartridge C1 QMA light (waters) conditioned with
potassium carbonate 0.5M Mix-Vial 1 mL acetonitrile 500 .mu.L
ethanol HPLC column Synergi Hydro-RP, 250*10 mm, 10 .mu.m 80 .ANG.,
Phenomenex HPLC solvent 60% ethanol, 40% ascorbate buffer (5 g/l
sodium ascorbate and 50 mg/l ascorbic acid) HPLC flow 6 mL/min
Start activity of 46.0 GBq [F-18]fluoride Product activity 19.4 GBq
Product radio- 99% purity (RCP) Radiochemical 42% (not corrected
for decay) yield
Example 5
Synthesis of
4-[(E)-2-(4-{2-[2-(2-[F-18]fluoroethoxy)ethoxy]-ethoxy}phenyl)vinyl]-N-me-
thylaniline Radiosynthesis on Tracerlab MX and Eckert&Ziegler
Purification Unit
[0137] The synthesis has been performed on GE TracerLab MX
synthesizer, purification of 4 has been performed on Eckert &
Ziegler Purification Unit. The filling of the injection loop of the
HPLC was controlled using the syringe of the MX module. The setup
of both automates and the results are summarized in the Table
below. [F-18]Fluoride was trapped on a QMA cartridge (C1). The
activity was eluted with a kryptofix mixture (from "V1") into the
reactor. The solvent was removed while heating under gentle
nitrogen stream and vacuum. Drying was repeated after addition of
acetonitrile (from "V2"). The solution of precursor 2a (from "V3")
was added to the dried residue and the mixture was heated for 10
min at 120.degree. C. After cooling to 40.degree. C., 2 mL 1.5M HCl
(from "V4") was added and solution was heated for 5 min at
110.degree. C.
[0138] The crude product mixture was diluted with 1.2 mL 2M NaOH
and 0.8 mL ammonium formate (1 M) from syringe "Si" and then
transferred to the HPLC vial ("Mix-Vial") in which 1 mL
acetonitrile (from "V2") and 0.5 mL ethanol (from "V5") are added
separately.
[0139] The average 6-7 mL mixture was transferred to a 30 mL
syringe which then pushed the totality of the volume into the 10 mL
sample injection loop of the semi-preparative HPLC. The mixture is
loaded to the semi-preparative HPLC column (Synergi Hydro-RP,
250.times.10 mm, Phenomenex). A mixture of 60% ethanol and 40%
ascorbate buffer was flushed through the column with 6 mL/min. The
product fraction at .apprxeq.9 min was collected for 50 sec
directly into the product vial containing 15 mL Formulation basis
(consisting of phosphate buffer, PEG400 and ascorbic acid).
Analytical HPLC of the final product showed excellent radiochemical
and chemical purity. No impurity higher than 0.5 .mu.g/mL was
quantified.
TABLE-US-00003 TABLE 3 Vial V1 22 mg kryptofix 7 mg potassium
carbonate 300 .mu.L acetonitrile 300 .mu.L water Vial V2 8 mL
acetonitrile Vial V3 8 mg precursor in 1.8 mL acetonitrile Vial V4
2 mL HCl 1.5M Vial V5 8 mL ethanol Syringe S1 1.2 mL NaOH 2.0M 800
.mu.L ammonium formate 1M Cartridge C1 QMA light (waters)
conditioned with potassium carbonate 0.5M HPLC column Synergi
Hydro-RP, 250* .times. 10 mm, 10 .mu.m 80 .ANG., Phenomenex HPLC
solvent 60% ethanol, 40% ascorbate buffer (5 g/l sodium ascorbate
and 50 mg/l ascorbic acid) HPLC flow 6 mL/min Start activity of
36.9 GBq [F-18]fluoride Product activity 14.2 GBq Product radio-
100% purity (RCP) Radiochemical 38% (not corrected for decay)
yield
Example 6
Synthesis of
4-[(E)-2-(4-{2-[2-(2-[F-18]fluoroethoxy)ethoxy]-ethoxy}phenyl)vinyl]-N-me-
thylaniline Radiosynthesis on Tracerlab MX and Eckert&Ziegler
Purification Unit
[0140] The synthesis has been performed on GE TracerLab MX
synthesizer, purification of 4 has been performed on Eckert &
Ziegler Purification Unit. The filling of the injection loop of the
HPLC was controlled by a fluid detector of the Eckert&Ziegler
Purification unit. The setup of both automates and the results are
summarized in the Table below. [F-18]Fluoride was trapped on a QMA
cartridge (C1). The activity was eluted with a kryptofix mixture
(from "V1") into the reactor.
[0141] The solvent was removed while heating under gentle nitrogen
stream and vacuum. Drying was repeated after addition of
acetonitrile (from "V2"). The solution of precursor (from "V3") was
added to the dried residue and the mixture was heated for 10 min at
120.degree. C. After cooling to 40.degree. C., 2 mL 1.5M HCl (from
"V4") was added and solution was heated for 5 min at 110.degree.
C.
[0142] The crude product mixture was diluted with 1.2 mL 2M NaOH
and 0.8 mL ammonium formate (1 M) from syringe "S1". 1 mL
acetonitrile (from "V2") and 0.5 mL ethanol (from "V5") are added
separately to the mixture and then transferred to the right syringe
of the GE TracerLab MX automate.
[0143] The mixture was 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 controlled the end
of the loading. The mixture was loaded to the semi-preparative HPLC
column (Synergi Hydro-RP, 250.times.10 mm, Phenomenex). A mixture
of 60% ethanol and 40% ascorbate buffer was flushed through the
column with 6 mL/min. The product fraction at .apprxeq.9 min was
collected directly during 50 sec into the product vial containing
15 mL Formulation basis (consisting of phosphate buffer, PEG400 and
ascorbic acid). Analytical HPLC of the final product showed
excellent radiochemical and chemical purity. No impurity higher
than 0.7 .mu.g/mL was quantified.
TABLE-US-00004 TABLE 4 Vial V1 22 mg kryptofix 7 mg potassium
carbonate 300 .mu.L acetonitrile 300 .mu.L water Vial V2 8 mL
acetonitrile Vial V3 8 mg precursor in 1.8 mL acetonitrile Vial V4
2 mL HCl 1.5M Vial V5 8 mL ethanol Syringe S1 1.2 mL NaOH 2.0M 800
.mu.L ammonium formate 1M Cartridge C1 QMA light (waters)
conditioned with potassium carbonate 0.5M HPLC column Synergi
Hydro-RP, 250* .times. 10 mm, 10 .mu.m 80 .ANG., Phenomenex HPLC
solvent 60% ethanol, 40% ascorbate buffer (5 g/l sodium ascorbate
and 50 mg/l ascorbic acid) HPLC flow 6 mL/min Start activity of
62.2 GBq [F-18]fluoride Product activity 24.8 GBq Product radio-
100% purity (RCP) Radiochemical 40% (not corrected for decay)
yield
Example 7
Influence of Purification Method on Radiochemical Purity
[0144] A series of
4-[(E)-2-(4-{2-[2-(2-[F-18]fluoroethoxy)ethoxy]-ethoxy}phenyl)vinyl]-N-me-
thylaniline syntheses was performed on two different synthesizers
(Eckert & Ziegler modular lab and GE tracerlab MX) as generally
described in the examples 1, 3-6. The radiolabelings have been
performed using 4-10 mg precursor 2a in acetonitrile as well as in
tert-amylalcohol/acetonitrile mixture at 100-120.degree. C. for
10-20 min. (in case of radiolabelings in tert-amylalcohol the
solvent was evaporated prior deprotection). The N-Boc protecting
group was removed by heating with HCl (1.5M-2M).
[0145] Crude product mixtures were individually purified by one of
the two methods A) or B).
Method A):
[0146] The crude product mixture obtained after deprotection is
neutralized with a mixture of 2M NaOH and 0.1M ammonium formate and
injected onto a semipreparative HPLC (e.g. column: Gemini C18,
10.times.250 mm, 5 .mu.m, Phenomenex; solvent: 70% acetonitrile,
30% ammonium formate buffer 0.1M with 5 mg/mL sodium ascorbate,
flow rate 3 mL/min). The product fraction is collected into a flask
containing approx. 160 mL water with 10 mg/mL sodium ascorbate. The
mixture is passed through a C18 cartridge (tC18 SepPak
environmental, Waters). The cartridge is washed with approx. 8-10
mL 20% EtOH in water (containing 10 mg/mL sodium ascorbate).
Finally, the product is eluted with 1.5 to 3 mL ethanol into a vial
containing 8.5 to 17 mL "Formulation basis" (comprising PEG400,
phosphate buffer and ascorbic acid).
Method B):
[0147] The crude product mixture obtained after deprotection is
neutralized with a mixture of 2M NaOH and 0.1M ammonium formate and
injected onto a semipreparative HPLC (column: e.g.: Gemini C18,
10.times.250 mm, 5 .mu.m, Phenomenex or Synergi Hydro-RP,
250.times.10 mm, 10 .mu.m 80 .ANG., Phenomenex or Synergi Hydro-RP,
250.times.10 mm, 4 .mu.M 80 .ANG., Phenomenex; solvent: 60-70%
ethanol, 40-30% ascorbate buffer .infin.5 mg/mL ascorbate; flow 3
mL/min or 4 mL/min or 6 mL/min). The product fraction is directly
collected into a vial containing "Formulation basis" (comprising
PEG400, phosphate buffer and ascorbic acid) to provide 10-24 mL of
the final Formulation. The peak-cutting time was adjusted in the
software to obtain a Formulation comprising 15% EtOH.
[0148] Every empty square (each one result for a synthesis
comprising a purification by method A, 110 experiments) and every
filled dot (each one result for a synthesis comprising a
purification by method B, 105 experiments) in FIG. 9 represents an
individual experiment for the manufacturing of
4-[(E)-2-(4-{2-[2-(2-[F-18]fluoroethoxy)ethoxy]-ethoxy}phenyl)vinyl]-N-me-
thylaniline. The tendency of radiochemical purity in correlation
with radioactivity of the final product is illustrated by linear
trendlines.
[0149] The radiochemical purity obtained after HPLC with
re-Formulation by SPE (method A) varies significantly (FIG. 9,
empty squares). Especially at higher radioactive levels (>20
GBq) the radiochemical purity often is even 95%.
[0150] In contrast, variability is much lower for method B).
Consistently high radiochemical purities of >95% were achieved
at activity levels of the product of greater than 50 GBq, and even
greater than 100 GBq (FIG. 9, filled dots).
Example 8
Synthesis of
4-[(E)-2-(6-{2-[2-(2-[F-18]fluoroethoxy)ethoxy]-ethoxy}pyridin-3-yl)vinyl-
]-N-methylaniline on Tracerlab FX.sub.N
##STR00015##
[0152] A Tracerlab FX.sub.N synthesizer has been adopted to the
"direct cut HPLC approach" (FIG. 4).
[0153] [F-18]Fluoride (10 GBq) was trapped on a QMA cartridge. The
activity was eluted with potassium
carbonate/kryptofix/acetonitrile/water mixture into the reactor.
The solvent was removed while heating under gentle nitrogen stream
and vacuum. Drying was repeated after addition of acetonitrile. A
solution of 8 mg 2b in 1.5 mL acetonitrile was added to the dried
residue and the mixture was heated for 10 min at 120.degree. C.
After cooling to 60.degree. C., 1 mL 1.5M HCl was added and the
reactor was heated at 110.degree. C. for 5 min. The crude product
was neutralized (1 mL 1M NaOH/ammonium formate), diluted (with 0.5
mL EtOH and 1.5 mL MeCN) and transferred to a semi-preparative HPLC
column (Synergy Hydro-RP, 250.times.10 mm, Phenomenex). A mixture
of 60% ethanol and 40% ascorbate buffer (5 g/l sodium ascorbate and
50 mg/l ascorbic acid, pH 7.0) was flushed through the column with
3 mL/min. The product fraction at .apprxeq.10 min (see FIG. 10) was
directly collected for 100 sec and mixed with 15 mL Formulation
basis (phosphate buffer, ascorbic acid, PEG400).
[0154] 4.2 GBq (42% not corrected for decay) were obtained in 61
min overall synthesis time. Radiochemical purity (determined by
HPLC, t.sub.R=3.42 min) was determined to be >99%.
Example 9
Synthesis of
4-[(E)-2-(4-{2-[2-(2-[F-18]fluoroethoxy)ethoxy]-ethoxy}phenyl)vinyl]-N-me-
thylaniline on Tracerlab FX.sub.N
##STR00016##
[0156] A Tracerlab FX.sub.N synthesizer have been adopted to the
"direct cut HPLC approach" (FIG. 4).
[0157] [F-18]Fluoride (6.85 GBq) was trapped on a QMA cartridge.
The activity was eluted with potassium
carbonate/kryptofix/acetonitrile/water mixture into the reactor.
The solvent was removed while heating under gentle nitrogen stream
and vacuum. Drying was repeated after addition of acetonitrile. A
solution of 8 mg 2c in 1.5 mL acetonitrile was added to the dried
residue and the mixture was heated for 10 min at 120.degree. C.
After cooling to 60.degree. C., the crude product was diluted with
4 mL HPLC eluent and transferred to a semi-preparative HPLC column
(Synergy Hydro-RP, 250.times.10 mm, Phenomenex). A mixture of 60%
ethanol and 40% ascorbate buffer (5 g/l sodium ascorbate and 50
mg/l ascorbic acid, pH 7.0) was flushed through the column with 3
mL/min. The product fraction at .apprxeq.12 min was directly
collected for 100 sec and mixed with 15 mL Formulation basis
(phosphate buffer, ascorbic acid, PEG400).
[0158] 2.54 GBq (37% not corrected for decay) were obtained in 53
min overall synthesis time. Radiochemical purity (determined by
HPLC, t.sub.R=3.78 min) was determined to be >99%.
DESCRIPTION OF THE FIGURES
[0159] FIG. 1 Setup of Tracerlab FX.sub.N for purification with
re-Formulation (adopted from tracerlab software)
[0160] FIG. 2 Chromatogramm of purification using Synergy column on
Eckert&Ziegler modular lab (Radioactivity channel)
[0161] FIG. 3 Analytical HPLC of radiolabeled product (top
radioactivity channel, bottom UV channel)
[0162] FIG. 4 Setup of Tracerlab FX.sub.N for purification without
re-Formulation (adopted from tracerlab software)
[0163] FIG. 5 Setup of Tracerlab MX (adopted from Coincidence FDG
software)
[0164] FIG. 6 Setup of Eckert&Ziegler purification unit
(adopted from Modual-Lab software)
[0165] FIG. 7 Schematic illustration of process and equipment for
manufacturing of F-18 labeled fluoropegylated (aryl/heteroaryl
vinyl)-phenyl methyl amines comprising three parts: A) Synthesis,
B) HPLC, C) Formulation; including (1) vials for reagents and
solvents, (2) a reaction vessel, (3) target line for F-18,
optionally gas lines, vacuum ect., (4) optionally fluid detector or
filter ect., (5) injection valve, (6) HPLC column, (7) valve for
peak cutting, (W) waste line(s), (8) vessel for collection/dilution
of HPLC fraction, (9) solvent vials for washing and elution, (10)
valve, (11) cartridge, e.g. C18 cartridge for trapping of the
product, (12) valve.
[0166] FIG. 8 Schematic illustration of process and equipment for
manufacturing of F-18 labeled fluoropegylated (aryl/heteroaryl
vinyl)-phenyl methyl amines comprising two parts: A) Synthesis, B)
HPLC; including (1) vials for reagents and solvents, (2) a reaction
vessel, (3) target line for F-18, optionally gas lines, vacuum
ect., (4) optionally fluid detector or filter ect., (5) injection
valve, (6) HPLC column, (7) valve for peak cutting.
[0167] FIG. 9 Influence of purification method on radiochemical
purity
[0168] FIG. 10 Chromatogramm of purification of
4-[(E)-2-(6-{2-[2-(2-[F-18]fluoro-ethoxy)ethoxy]ethoxy}pyridin-3-yl)vinyl-
]-N-methylaniline on Eckert&Ziegler modular lab (Radioactivity
channel)
* * * * *