U.S. patent application number 14/428876 was filed with the patent office on 2015-09-03 for imaging agents.
The applicant listed for this patent is THE REGENTS OF THE UNIVERSITY OF MICHIGAN. Invention is credited to Keun-Sam Jang, Yong-Woon Jung, David M. Raffel.
Application Number | 20150246142 14/428876 |
Document ID | / |
Family ID | 50388933 |
Filed Date | 2015-09-03 |
United States Patent
Application |
20150246142 |
Kind Code |
A1 |
Raffel; David M. ; et
al. |
September 3, 2015 |
IMAGING AGENTS
Abstract
Provided herein is technology relating to imaging agents and
particularly, but not exclusively, to methods of manufacturing
fluorine-18-labeled phenethylguanidines and uses thereof.
Inventors: |
Raffel; David M.; (Ann
Arbor, MI) ; Jung; Yong-Woon; (Ann Arbor, MI)
; Jang; Keun-Sam; (Ann Arbor, MI) |
|
Applicant: |
Name |
City |
State |
Country |
Type |
THE REGENTS OF THE UNIVERSITY OF MICHIGAN |
Ann Arbor |
MI |
US |
|
|
Family ID: |
50388933 |
Appl. No.: |
14/428876 |
Filed: |
September 25, 2013 |
PCT Filed: |
September 25, 2013 |
PCT NO: |
PCT/US2013/061681 |
371 Date: |
March 17, 2015 |
Related U.S. Patent Documents
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Application
Number |
Filing Date |
Patent Number |
|
|
61705477 |
Sep 25, 2012 |
|
|
|
Current U.S.
Class: |
424/1.89 ;
544/11; 544/12; 544/179; 544/2; 544/235; 544/237; 544/283; 544/5;
544/50; 544/63; 544/65; 544/66; 544/90; 546/145; 548/122; 548/123;
548/124; 548/126; 548/257; 548/309.7; 548/362.5; 548/505;
564/237 |
Current CPC
Class: |
C07B 2200/05 20130101;
C07C 277/08 20130101; C07C 279/06 20130101; C07B 59/001 20130101;
A61K 31/155 20130101; C07C 279/08 20130101; A61K 51/04 20130101;
C07C 277/06 20130101; C07B 59/002 20130101 |
International
Class: |
A61K 51/04 20060101
A61K051/04; C07B 59/00 20060101 C07B059/00; C07C 279/08 20060101
C07C279/08 |
Goverment Interests
STATEMENT REGARDING FEDERALLY SPONSORED RESEARCH OR DEVELOPMENT
[0002] This invention was made with government support under
HL079540 awarded by the National Institutes of Health. The
government has certain rights in the invention.
Claims
1.-205. (canceled)
206. A composition comprising an .sup.18F-labeled
arylalkylguanidine having a structure according to ##STR00063## or
a salt, a free base, or a combination thereof, wherein: a) R.sub.1,
R.sub.2, R.sub.3, R.sub.4, and R.sub.5 are independently selected
from the group consisting of .sup.18F-labeled, .sup.19F, hydrogen,
halogen, hydroxyl, guanyl, alkoxy, haloalkoxy, .sup.18F-labeled
alkoxy, alkyl, haloalkyl, .sup.18F-labeled alkyl, amine, and an
amine comprising a protecting group; b) n is equal to 0 or 2; c) Z
is independently selected from the group comprising hydrogen,
hydroxyl, alkoxy, haloalkoxy, .sup.18F-labeled alkoxy, halogen,
amino, alkyl, haloalkyl, and .sup.18F-labeled alkyl; and d)
R.sub.6, R.sub.7, R.sub.8, and R.sub.9 are independently selected
from the group consisting of hydrogen, carbamate, cyclic carbamate,
amide, cyclic amide, and a nitrogen-protecting group.
207. The composition of claim 206 wherein R.sub.6 is a hydrogen,
R.sub.7 is a hydrogen, R.sub.8 is a hydrogen, and/or R.sub.9 is a
hydrogen.
208. The composition of claim 206 further comprising a biologically
compatible carrier.
209. The composition of claim 206 further comprising a
physiological buffered saline solution or a physiological buffered
sodium acetate carrier.
210. The composition of claim 206 comprising 0.1 to 100 mCi of
labeled radiotracing agent.
211. The composition of claim 206 comprising 50 to 500 mCi of
labeled radiotracing agent.
212. A composition comprising an .sup.18F-labeled
aryl-Y-alkylguanidine having a structure according to ##STR00064##
or a salt, a free base, or a combination thereof, wherein: a)
R.sub.1, R.sub.2, R.sub.3, R.sub.4, and R.sub.5 are independently
selected from the group consisting of .sup.18F-labeled, .sup.19F,
hydrogen, halogen, hydroxyl, guanyl, alkoxy, haloalkoxy,
.sup.18F-labeled alkoxy, alkyl, haloalkyl, .sup.18F-labeled alkyl,
amine, and an amine comprising a protecting group; b) Y is
independently selected from the group comprising O, S or NH. c)
R.sub.6, R.sub.7, R.sub.8, and R.sub.9 are independently selected
from the group consisting of hydrogen, carbamate, cyclic carbamate,
amide, cyclic amide, and a nitrogen-protecting group; and d) n is
equal to 1, 2 or 3.
213. The composition of claim 212 wherein R.sub.6 is a hydrogen,
R.sub.7 is a hydrogen, R.sub.8 is a hydrogen, and/or R.sub.9 is a
hydrogen.
214. The composition of claim 212 further comprising a biologically
compatible carrier.
215. The composition of claim 212 further comprising a
physiological buffered saline solution or a physiological buffered
sodium acetate carrier.
216. The composition of claim 212 comprising 0.1 to 100 mCi of
labeled radiotracing agent.
217. The composition of claim 212 comprising 50 to 500 mCi of
labeled radiotracing agent.
218. A composition comprising an .sup.18F-labeled
heteroarylalkylguanidine having a structure according to:
##STR00065## or a salt, a free base, or a combination thereof,
wherein: e) R.sub.1, R.sub.2, R.sub.3, and R.sub.4 are
independently selected from the group consisting of
.sup.18F-labeled, .sup.19F, hydrogen, halogen, hydroxyl, guanyl,
alkoxy, haloalkoxy, .sup.18F-labeled alkoxy, alkyl, haloalkyl,
.sup.18F-labeled alkyl, amine, and an amine comprising a protecting
group; f) L, M, N and Q are independently selected from the group
comprising CH.sub.2, CH, O, N, NH, S, CO, alkyl, haloalkyl, alkoxy,
haloalkoxy, .sup.18F-labeled alkyl, and .sup.18F-alkoxy; g)
R.sub.5, R.sub.6, R.sub.7, and R.sub.8 are independently selected
from the group consisting of hydrogen, carbamate, cyclic carbamate,
amide, cyclic amide, and a nitrogen-protecting group; and h) n is
equal to 0, 1, 2 or 3.
219. The composition of claim 218 wherein R.sub.5 is a hydrogen,
R.sub.6 is a hydrogen, R.sub.7 is a hydrogen, and/or R.sub.8 is a
hydrogen.
220. The composition of claim 218 further comprising a biologically
compatible carrier.
221. The composition of claim 218 further comprising a
physiological buffered saline solution or a physiological buffered
sodium acetate carrier.
222. The composition of claim 218 comprising 0.1 to 100 mCi of
labeled radiotracing agent.
223. The composition of claim 218 comprising 50 to 500 mCi of
labeled radiotracing agent.
Description
[0001] This application claims priority to U.S. Patent Application
Ser. No. 61/705,477 filed on Sep. 25, 2012, which is incorporated
herein by reference in its entirety for all purposes.
FIELD OF TECHNOLOGY
[0003] Provided herein is technology relating to imaging agents and
particularly, but not exclusively, to methods of producing
fluorine-18-labeled guanidine compounds.
BACKGROUND
[0004] Medical radionuclide imaging (e.g., nuclear medicine) is a
key component of modern medical practice. This methodology involves
the administration, typically by injection, of tracer amounts of a
radioactive substance (e.g., radiotracer agents, radiotherapeutic
agents, and radiopharmaceutical agents), which subsequently
localize in the body in a manner dependent on the physiologic
function of the organ or tissue system being studied. The
radiotracer emissions, most commonly gamma photons, are imaged with
a detector outside the body, creating a map of the radiotracer
distribution within the body. When interpreted by an appropriately
trained physician, these images provide information of great value
in the clinical diagnosis and treatment of disease. Typical
applications of this technology include detection of coronary
artery disease (e.g., thallium scanning) and detection of cancerous
involvement of bones (e.g., bone scanning). The overwhelming bulk
of clinical radionuclide imaging is performed using gamma emitting
radiotracers and detectors known as "gamma cameras".
[0005] Recent advances in diagnostic imaging, such as magnetic
resonance imaging (MRI), computerized tomography (CT), single
photon emission computerized tomography (SPECT), and positron
emission tomography (PET), have made a significant impact in
cardiology, neurology, oncology, and radiology. Although these
diagnostic methods employ different techniques and yield different
types of anatomic and functional information, this information is
often complementary in the diagnostic process.
[0006] Imaging agents are generally classified as either being
diagnostic or therapeutic in their application. Although diagnostic
imaging agents have historically been a mainstay in the nuclear
pharmacy industry, during the past decade there has been increased
interest in the development and use of radioactive imaging agents
for radiotherapy. This shift in focus has been elicited primarily
from research involving combining radioactive isotopes with
sophisticated molecular carriers. Because of radiation's damaging
effect on tissues, it is important to target the biodistribution of
radiopharmaceuticals as accurately as possible. Generally speaking,
PET uses imaging agents labeled with positron-emitters such as
.sup.18F, .sup.11C, .sup.13N, .sup.15O, .sup.75Br, .sup.76Br, and
.sup.124I; SPECT uses imaging agents labeled with
single-photon-emitters such as .sup.201Tl, .sup.99mTc, .sup.123I,
and .sup.131I.
[0007] In the art, glucose-based and amino acid-based compounds
have been used as imaging agents. Amino acid-based compounds are
more useful in analyzing tumor cells due to their faster uptake and
incorporation into protein synthesis. Of the amino acid-based
compounds, .sup.11C- and .sup.18F-containing compounds have been
used with success. .sup.11C-containing radiolabeled amino acids
suitable for imaging include, for example, L-[1-.sup.11C]leucine
(Keen et al. J. Cereb. Blood Flow Metab. 1989 (9):429-45; herein
incorporated by reference in its entirety), L-[1-.sup.11C]tyrosine
(Wiesel et al. J. Nucl. Med. 1991 (32):2041-49; herein incorporated
by reference in its entirety), L-[methyl-.sup.11C]methionine (Comar
et al. Eur. J. Nucl. Med. 1976 (1):11-14; herein incorporated by
reference in its entirety) and L-[1-.sup.11C]methionine (Bolster et
al. Appl. Radiat. Isot. 1986 (37):1069-70; herein incorporated by
reference in its entirety).
[0008] PET involves the detection of gamma rays in the form of
annihilation photons from short-lived positron emitting radioactive
isotopes including but not limited to .sup.18F with a half-life of
approximately 110 minutes, .sup.11C with a half-life of
approximately 20 minutes, .sup.13N with a half-life of
approximately 10 minutes, and .sup.15O with a half-life of
approximately 2 minutes, using the coincidence method.
[0009] For PET imaging studies of cardiac sympathetic innervation,
carbon-11 (.sup.11C) labeled compounds such as
[.sup.11C]meta-hydroxyephedrine (HED) are frequently used at major
PET centers that have in-house cyclotrons and radiochemistry
facilities. However, the nuclear medicine market has recently seen
a substantial increase in stand-alone PET imaging centers that do
not have cyclotrons and that primarily use
2-[.sup.18F]fluoro-2-deoxy-D-glucose (FDG) for PET imaging of
cancerous tumors.
[0010] SPECT, on the other hand, uses longer-lived isotopes
including but not limited to .sup.99mTc with a half-life of
approximately 6 hours and .sup.201Tl with a half-life of
approximately 74 hours. However, the resolution in present SPECT
systems is lower than that presently available in PET systems.
[0011] Radio-iodinated meta-iodobenzylguanidine (MIBG) is a
radiotracing agent that is used, for example, in nuclear medicine
imaging studies of sympathetic nerve fibers in the human heart.
Studies with MIBG allow clinicians to map the regional distribution
of nerve fibers in the heart using imaging devices found in all
nuclear medicine clinics. MIBG is also used for diagnostic imaging
and radiotherapy of adrenergic tumors, such as neuroblastoma and
pheochromocytoma.
[0012] New compounds that find use as imaging agents within nuclear
medicine applications (e.g., PET imaging and SPECT imaging) have
been described: for example, fluorine-18-labeled
phenethylguanidines. See, e.g., U.S. Pat. No. 7,534,418,
incorporated herein by reference in its entirety for all
purposes.
[0013] While useful, introducing fluorine-18 into a phenyl ring
moiety at high specific activities is a notoriously challenging
radiolabeling task, especially in electron-rich aromatic systems.
In the last 10 years, the use of diaryliodium salt precursors as a
one-step method of introducing fluorine-18 into ring structures
with high radiochemical yields has received considerable attention.
Although this method has been used to prepare small model compounds
with relatively simple structures, as the structures of the
compounds being radiolabeled become more complex, radiochemical
yields drop substantially. Accordingly, a need exists for methods
of preparing fluorine-18 labeled phenethylguanidines and related
compounds with high specific activity to make practical the
production of such compounds on a commercial scale.
SUMMARY
[0014] Accordingly, provided herein is technology related to
preparing ring-fluorinated guanidine radiotracers at high specific
activities and high yields. As provided herein, a first step in
particular embodiments of the technology uses a novel
diaryliodonium salt precursor to introduce fluorine-18 into the
ring structure, followed by removal of a N-Boc protecting group to
yield a radiolabeled 4-[.sup.18F]fluoro-meta-tyramine derivative in
which the meta-hydroxy group remains protected by a benzyl group.
This intermediate is then converted from a primary amine to a
guanidine using N--N'-diBoc-5-chlorobenzotriazole. A further step
deprotects the meta-hydroxy group to yield
4-[.sup.18F]fluoro-meta-hydroxy-phenethylguanidine
([.sup.18F]4F-MHPG; in some contexts, referred to by the name
4-[.sup.18F]-MHPG).
[0015] As such, the technology relates to new methods of producing
radioactive compounds; in particular, the methods relate to using a
diaryliodium salt precursor to introduce fluorine-18 into the
structure of a radiolabeled phenethylguanidine and related
compounds and, in some embodiments, the methods relate to preparing
a .sup.18F-labeled primary amine intermediate followed by
conversion to a guanidine. As provided herein, the methods are used
to produce an exemplary novel compound, [.sup.18F]4F-MHPG. The
technology is contemplated to encompass related and generalized
structures defining a unified set of novel diaryliodium salt
precursors that are used to prepare .sup.18F-labeled
phenethylguanidines as disclosed in, e.g., U.S. Pat. No. 7,534,418,
which is incorporated by reference in its entirety for all
purposes.
[0016] Accordingly, provided herein is technology related in one
aspect to compositions comprising an .sup.18F-labeled
phenethylguanidine having a structure according to
##STR00001##
or a salt, a free base, or a combination thereof, wherein R.sub.1,
R.sub.2, R.sub.3, R.sub.4, and R.sub.5 are independently selected
from the group consisting of .sup.18F-labeled, .sup.19F, hydrogen,
halogen, hydroxyl, guanyl, alkoxy, haloalkoxy, .sup.18F-labeled
alkoxy, alkyl, haloalkyl, .sup.18F-labeled alkyl, amine, and an
amine comprising one or more protecting groups (e.g., a protected
amine); R.sub.6 and R.sub.7 are independently selected from the
group consisting of hydrogen, hydroxyl, alkoxy, haloalkoxy,
.sup.18F-labeled alkoxy, halogen, amino, alkyl, haloalkyl, and
.sup.18F-labeled alkyl; and R.sub.8, R.sub.9, R.sub.10, and
R.sub.11 are independently selected from the group consisting of
hydrogen, carbamate, cyclic carbamate, amide, cyclic amide, and a
nitrogen-protecting group. In some embodiments of the composition,
R.sub.8 is a hydrogen, R.sub.9 is a hydrogen, R.sub.10 is a
hydrogen, and/or R.sub.11 is a hydrogen.
[0017] Also provided are embodiments related to compositions
comprising a .sup.18F-labeled phenethylguanidine, wherein the
.sup.18F-labeled phenethylguanidine is produced by a method
comprising radiofluorinating an iodonium salt with an
[.sup.18F]fluoride ion source, e.g., as described by Reaction
1:
##STR00002##
[0018] In some embodiments, the iodonium salt has a structure
according to
##STR00003##
or is a salt, a free base, or a combination thereof, wherein
R.sub.1, R.sub.2, R.sub.3, R.sub.4, and R.sub.5 are independently
selected from the group consisting of R.sub.12--I, hydrogen,
halogen, hydroxyl, guanyl, alkoxy, haloalkoxy, alkyl, haloalkyl,
amine, and an amine comprising one or more protecting groups (e.g.,
a protected amine); R.sub.6 and R.sub.7 are independently selected
from the group consisting of hydrogen, hydroxyl, alkoxy,
haloalkoxy, halogen, amino, alkyl, and haloalkyl; and R.sub.8,
R.sub.9, R.sub.10, and R.sub.11 are independently selected from the
group consisting of hydrogen, carbamate, cyclic carbamate, amide,
cyclic amide, and a nitrogen-protecting group. Furthermore, in some
embodiments, R.sub.12 is a phenyl ring or a heterocyclic ring
comprising hydrogen, hydroxyl, alkyl, halogen, alkoxy, carbonyl,
cyano, and/or a nitro group. Some embodiments, moreover, comprise a
.sup.18F-labeled phenethylguanidine wherein R.sub.12 comprises a
solid-support bound linker, e.g.,
##STR00004##
[0019] In some embodiments, the [.sup.18F]fluoride ion source is a
no-carrier-added [.sup.18F]fluoride ion source. For example, in
some embodiments, the no-carrier-added [.sup.18F]fluoride ion
source is selected from the group consisting of potassium
fluoride/Kryptofix[2,2,2], cesium fluoride, tetraalkylammonium
fluoride, and a solid phase fluoride. In some embodiments, X- is a
counter ion selected from the group consisting of halide, sulfate,
formate, bromate, tosylate, trifluoracetate, triflate, mesylate,
hexaflate, acetate, ascorbate, benzoate, and phosphate.
Compositions are also provided that furthermore comprise a free
radical scavenger, e.g., as a component of a reaction in which the
compositions and compounds according to the technology are made.
Examples of free radical scavengers include, in some embodiments,
2,2,6,6-tetramethylpiperidine-N-oxide, 4-aminobenzoic acid,
1,1-diphenylethylene, galvinoxyl, gentisic acid, hydroquinone,
thiophenol, DL-alpha-tocopherol, and
2,6-di-tert-butyl-4-methylphenol (BHT). In some embodiments, the
compositions provided further comprise water. In some embodiments,
a reaction is heated, e.g., in some embodiments, compositions are
produced according to a method that comprises heating or microwave
irradiation of a reaction vessel holding the iodonium salt and the
[.sup.18F]fluoride ion source.
[0020] In another aspect, the technology relates to compositions
comprising an .sup.18F-labeled phenethylguanidine, wherein the
.sup.18F-labeled phenethylguanidine is produced by a method
comprising radiofluorinating an iodonium salt of phenethylamine
with a [.sup.18F]fluoride ion source to produce an
[.sup.18F]-labeled phenethylamine; and coupling the
[.sup.18F]-labeled phenethylamine with a guanidinating reagent to
produce a [.sup.18F]-labeled phenethylguanidine, e.g., as described
by Reaction 2:
##STR00005##
[0021] For example, in some embodiments, the iodonium salt of
phenethylamine
##STR00006##
or a salt, free base, or combination thereof, wherein R.sub.1',
R.sub.2', R.sub.3', R.sub.4', and R.sub.5' are independently
selected from the group consisting of R.sub.10'--I, hydrogen,
halogen, hydroxyl, guanyl, alkoxy, haloalkoxy, alkyl, haloalkyl,
amine, and an amine comprising one or more protecting groups (e.g.,
a protected amine); R.sub.6' and R.sub.7' are independently
selected from the group consisting of hydrogen, hydroxyl, alkoxy,
haloalkoxy, halogen, amino, alkyl, and haloalkyl; and R.sub.8' and
R.sub.9' are independently selected the group from consisting of
hydrogen, carbamate, cyclic carbamate, amide, cyclic amide, and
nitrogen-protecting group.
[0022] The technology is not limited in the guanidinating agent
that can be used: for example, in some embodiments, the
guanidinating reagent is selected from the group consisting of a
cyamide, cyanobromide/ammonia, an S-alkylisothiouronium salt, a
carboimide, a chloroformamidine, a dichloroisocyanide, an
aminoimnomethanesulfonic acid, O-methylisourea hydrogen sulfate,
1H-pyrazole-1-carboxamidine hydrochloride,
benzotriazole-1-carboxamidinium tosylate,
1H-pyrazole-1-[N,N'-Bis(tert-butoxy/benzyloxycarbonyl)]-carboxamidine,
N,N'-bis(tert-butoxy/benzyloxycarbonyl)-N''-trifly guanidine,
N,N'-bis(tert-butoxy/benzyloxycarbonyl)-2-methyl-2-thiopseudourea,
N,N'-bis(tert-butoxy/benzyloxycarbonyl)-thiourea,
N,N'-bis(tert-butoxy/benzyloxycarbonyl)-carboimide, and
N,N'-bis(tert-butoxy/benzyloxycarbonyl)-1H-benzotriazole-1-carboxamidine.
[0023] Embodiments of compositions are provided wherein R.sub.10'
is a phenyl ring or a heterocyclic ring comprising a hydrogen,
hydroxyl, alkyl, halogen, alkoxy, carbonyl, cyano, and/or a nitro
group. In addition, in some embodiments, R.sub.10' comprises a
solid support linker, e.g.,
##STR00007##
Certain embodiments comprise a composition produced using an
iodonium salt of phenethylamine that is an [.sup.18F]-labeled
phenethylamine derivative according to a structure
##STR00008##
In some embodiments, the [.sup.18F]-labeled phenethylguanidine has
the structure
##STR00009##
In some embodiments, the [.sup.18F]fluoride ion source is a
no-carrier-added [.sup.18F]fluoride ion source, e.g., a potassium
fluoride/Kryptofix [2,2,2], cesium fluoride, and/or
tetraalkylammonium fluoride. In some embodiments, X- is a counter
ion selected from the group consisting of halide, sulfate, formate,
borate, tosylate, trifluoroacetate, triflate, mesylate, hexaflate,
acetate, ascorbate, benzoate, and phosphate. In some embodiments,
the compositions comprise a free radical scavenger, e.g., as a
component of a reaction to produce embodiments of the technology
comprising compositions and compounds. Exemplary free radical
scavengers include 2,2,6,6-tetramethylpiperidine-N-oxide,
4-aminobenzoic acid, 1,1-diphenylethylene, galvinoxyl, gentisic
acid, hydroquinone, thiophenol, DL-alpha-tocopherol, and
2,6-di-tert-butyl-4-methylphenol (BHT). In some embodiments, the
compositions comprise water. In some embodiments, a reaction is
heated, e.g., in some embodiments, compositions according to the
technology are produced by a method that comprises heating or
microwave irradiation of a reaction vessel holding the iodonium
salt and the [.sup.18F]fluoride ion source.
[0024] In another aspect, embodiments of the technology comprise
compositions wherein the iodonium salt is produced by a method
comprising reacting a first compound having the structure
##STR00010##
with a second compound that is R.sub.12-Koser's Reagent, e.g., as
described by Reaction 3:
##STR00011##
[0025] In some embodiments, R.sub.1, R.sub.2, R.sub.3, R.sub.4, and
R.sub.5, are independently selected from the group consisting of
X.sub.3Sn, hydrogen, halogen, hydroxyl, guanyl, alkoxy, haloalkoxy,
alkyl, haloalkyl, amine, and an amine comprising one or more
protecting groups (e.g., a protected amine); R.sub.6 and R.sub.7
are independently selected from the group consisting of hydrogen,
hydroxyl, alkoxy, haloalkoxy, halogen, amino, alkyl, and haloalkyl;
R.sub.8, R.sub.9, R.sub.10, and R.sub.11 are independently selected
the group from hydrogen, carbamate, cyclic carbamate, amide, cyclic
amide, and nitrogen-protecting group; and X is an alkyl group.
Furthermore, in some embodiments, R.sub.12 is a phenyl ring or a
heterocyclic ring comprising a hydrogen, hydroxyl, alkyl, halogen,
alkoxy, carbonyl, cyano, and/or a nitro group.
[0026] In another aspect, embodiments comprise compositions (e.g.,
a reaction pathway intermediate) that are produced by a method
comprising reacting a compound having the structure
##STR00012##
e.g., according to the reaction described by Reaction 4:
##STR00013##
[0027] In some embodiments, R.sub.1, R.sub.2, R.sub.3, R.sub.4, and
R.sub.5, are independently selected from the group consisting of
halogen (e.g., iodo, bromo), hydrogen, hydroxyl, guanyl, alkoxy,
haloalkoxy, alkyl, haloalkyl, amine, and an amine comprising one or
more protecting groups (e.g., a protected amine); R.sub.6 and
R.sub.7 are independently selected from the group consisting of
hydrogen, hydroxyl, alkoxy, haloalkoxy, halogen, amino, alkyl, and
haloalkyl; R.sub.8, R.sub.9, R.sub.10, and R.sub.11 are
independently selected the group consisting of hydrogen, carbamate,
cyclic carbamate, amide, cyclic amide, and nitrogen-protecting
group; and X is an alkyl group.
[0028] In another aspect, the iodonium salt of phenethylamine is
produced by a method comprising reacting a first compound having
the structure
##STR00014##
with a second compound that is R.sub.10'-Koser's Reagent, e.g.,
according to a reaction described in Reaction 5:
##STR00015##
Furthermore, in some embodiments, R.sub.1', R.sub.2', R.sub.3',
R.sub.4', and R.sub.5' are independently selected from the group
consisting of X.sub.3Sn, hydrogen, halogen, hydroxyl, guanyl,
alkoxy, haloalkoxy, alkyl, haloalkyl, amine, and an amine
comprising one or more protecting groups (e.g., a protected amine);
R.sub.6' and R.sub.7' are independently selected from the group
consisting of hydrogen, hydroxyl, alkoxy, haloalkoxy, halogen,
amino, alkyl, and haloalkyl: R.sub.8' and R.sub.9' are
independently selected the group from hydrogen, carbamate, cyclic
carbamate, amide, cyclic amide, and nitrogen-protecting group; and
X is an alkyl group. In some embodiments, R.sub.10' is a phenyl
ring or a heterocyclic ring comprising hydrogen, hydroxyl, alkyl,
halogen, alkoxy, carbonyl, cyano, and/or a nitro group.
[0029] Some aspects of the technology are related to embodiments of
compositions wherein a compound is produced by a method comprising
reacting a compound having the structure
##STR00016##
In some embodiments, R.sub.1', R.sub.2', R.sub.3', R.sub.4', and
R.sub.5' are independently selected from the group consisting of
iodo, bromo, hydrogen, halogen, hydroxyl, guanyl, alkoxy,
haloalkoxy, alkyl, haloalkyl, amine, and an amine comprising one or
more protecting groups (e.g., a protected amine); R.sub.6' and
R.sub.7' are independently selected from the group consisting of
hydrogen, hydroxyl, alkoxy, haloalkoxy, halogen, amino, alkyl, and
haloalkyl; R.sub.8' and R.sub.9' are independently selected from
the group consisting of hydrogen, carbamate, cyclic carbamate,
amide, cyclic amide, and nitrogen-protecting group.
[0030] The compounds and compositions provided by the technology
find use in imaging a tissue, cell, organ, e.g., in a subject.
Accordingly, the technology relates, in some embodiments, to
methods of imaging comprising contacting a tissue to be imaged with
an .sup.18F-labeled phenethylguanidine, or salt or derivative
thereof, and imaging the tissue. In some embodiments, the tissue is
selected from the group consisting of heart and adrenal medulla.
Furthermore, some embodiments provide that the tissue is suspected
of comprising a cancer. In some specific embodiments, the imaging
is positron emission tomography (PET).
[0031] Furthermore, the technology relates to the use of an
.sup.18F-labeled phenethylguanidine, or salt, free base, or
derivative thereof, to image a subject. In additional aspects, the
technology relates to embodiments of methods for manufacturing an
.sup.18F-labeled phenethylguanidine having a structure according
to
##STR00017##
or a salt, a free base, or a combination thereof, comprising
radiofluorinating an iodonium salt with an [.sup.18F]fluoride ion
source. In another aspect, the technology relates to embodiments of
methods for manufacturing an [.sup.18F]-labeled phenethylguanidine
having a structure according to
##STR00018##
or a salt, a free base, or a combination thereof, comprising the
steps of radiofluorinating an iodonium salt of phenethylamine with
a [.sup.18F]fluoride ion source to produce an [.sup.18F]-labeled
phenethylamine; and coupling the [.sup.18F]-labeled phenethylamine
with a guanidinating reagent to produce a [.sup.18F]-labeled
phenethylguanidine. In related embodiments, the technology provides
an .sup.18F-labeled phenethylguanidine, or salt, free base, or
derivative thereof, for use as an imaging agent. Moreover, in some
embodiments, provided herein is an .sup.18F-labeled
phenethylguanidine, or salt, free base, or derivative thereof, for
use as an imaging agent for the diagnosis of cancer or
cardiovascular disease.
[0032] The labeling technology is not limited to labeled
phenethylguanidines. In addition, the technology is applicable to
produce .sup.18F-labeled arylalkylguanidines, .sup.18F-labeled
aryl-Y-alkylguanidines, and/or .sup.18F-labeled
heteroarylalkylguanidines. For example, the technology relates to
embodiments of arylalkylguanidine compounds having a general
structure:
##STR00019##
embodiments of aryl-Y-alkylguanidine compounds having the
structure:
##STR00020##
and embodiments of heteroarylalkylguanidine compounds having the
structures:
##STR00021## [0033] n=0, 1, 2 or 3 [0034] L, M, N or Q=CH.sub.2,
CH, O, N, NH, S, CO, alkyl, haloalkyl, alkoxy, haloalkoxy,
.sup.18F-labeled alkyl or .sup.18F-labeled alkoxy
##STR00022##
[0035] Provided herein are technologies related to methods of
producing and/or manufacturing .sup.18F-labeled
arylalkylguanidines, .sup.18F-labeled aryl-Y-alkylguanidines,
and/or .sup.18F-labeled heteroarylalkylguanidines, e.g., for use as
imaging agents, e.g., in PET imaging. For example, some embodiments
provide methods in which an .sup.18F-labeled arylalkylguanidine is
produced from an iodonium salt precursor by a single step reaction
in solution, e.g.,
##STR00023##
The technology provides related embodiments in which an
arylalkylguanidine is produced from an iodonium salt precursor in a
single step using a linker, e.g.,
##STR00024##
and embodiments in which an arylalkylguanidine is produced from an
iodonium salt precursor in solution by the two-step reaction:
##STR00025##
[0036] In addition, some embodiments provide methods in which an
.sup.18F-labeled aryl-Y-alkylguanidine is produced from an iodonium
salt precursor by a single step reaction in solution, e.g.,
##STR00026##
The technology provides related embodiments in which an
aryl-Y-alkylguanidine is produced from an iodonium salt precursor
in a single step using a linker, e.g.,
##STR00027##
and embodiments in which an aryl-Y-alkylguanidine is produced from
an iodonium salt precursor in solution by the two-step
reaction:
##STR00028##
[0037] Finally, some embodiments provide methods in which an
.sup.18F-labeled heteroarylalkylguanidine is produced from an
iodonium salt precursor by a single step reaction in solution,
e.g.,
##STR00029##
[0038] The technology provides related embodiments in which a
heteroarylalkylguanidine is produced from an iodonium salt
precursor in a single step using a linker, e.g.,
##STR00030##
and embodiments in which a heteroarylalkylguanidine is produced
from an iodonium salt precursor in solution by the two-step
reaction:
##STR00031##
[0039] Additional embodiments will be apparent to persons skilled
in the relevant art based on the teachings contained herein.
BRIEF DESCRIPTION OF THE DRAWINGS
[0040] These and other features, aspects, and advantages of the
present technology will become better understood with regard to the
following drawings:
[0041] FIG. 1 shows a reaction scheme depicting a radiosynthetic
method for preparing .sup.18F-labeled phenethylguanidines using
diaryliodium salt precursors containing a phenethylguanidine
moiety.
[0042] FIG. 2 shows reaction schemes depicting radiosynthetic
methods for preparing
4-[.sup.18F]fluoro-meta-hydroxyphenethylguanidine using a
diaryliodium salt precursor containing a protected phenethylamine
moiety. This method can be generalized to prepare many other
.sup.18F-phenethylguanidine structures, such as those described in
U.S. Pat. No. 7,534,418. FIG. 2A and FIG. 2B show two exemplary
reaction schemes related to the technology provided herein, e.g.,
FIG. 2 shows an embodiment for the automated radiosynthesis of
4-[.sup.18F]fluoro-meta-hydroxyphenethylguanidine
([.sup.18F]4F-MHPG, compound 1).
[0043] FIG. 3 is a plot showing the kinetics of [.sup.11C]4F-MHPG
and [.sup.18F]4F-MHPG in isolated rat hearts.
[0044] FIG. 4 is a plot showing reverse-phase HPLC analysis of
[.sup.18F]4F-MHPG and its metabolites in rhesus macaque plasma.
FIG. 4A shows a representative HPLC trace for a blood sample drawn
at t=2 min after tracer injection. FIG. 4B shows that 100% of the
compound was in the sulfur-conjugated form after in vitro
incubation of the parent compound with a monkey liver cytosol
fraction and 3'-phospho-adenosine-5'phosphosulfate (PAPS). FIG. 5C
shows the structure of the sulfur conjugated form.
[0045] FIG. 5 is a plot showing metabolic breakdown of
[.sup.18F]4F-MHPG in the plasma of a rhesus macaque monkey.
[0046] FIG. 6 is a plot showing kinetics of [.sup.18F]4F-MHPG in
whole blood (lower trace) and left ventricle (top trace) in a
rhesus macaque monkey.
[0047] FIG. 7 is a diagram showing a compartmental model used to
analyze the myocardial kinetics of [.sup.18F]4F-MHPG.
[0048] FIG. 8 is a series of plots showing compartmental modeling
of [.sup.18F]4F-MHPG kinetics in monkeys for control (left),
moderate desipramine (DMI) dose blockade of cardiac NET (middle),
and high DMI dose (right).
[0049] FIG. 9 is a plot showing dose-response curves of net uptake
constants K.sub.i (ml/min/g) derived from either kinetic
compartmental modeling (circles) or Patlak graphical analysis
(triangles) of [.sup.18F]4F-MHPG kinetics in rhesus macaque
monkeys.
[0050] FIG. 10 is a plot showing Patlak analysis of the myocardial
kinetics of [.sup.18F]4F-MHPG kinetics in rhesus macaque
monkey.
[0051] FIG. 11 is a plot showing Patlak analysis of the myocardial
kinetics of [.sup.18F]4F-MHPG in rhesus macaque monkey.
[0052] FIG. 12 is a plot showing rapid neuronal uptake of
.sup.11C-labeled phenoxyethylguanidines in an isolated rat heart
model.
[0053] FIG. 13 is a plot showing the rapid neuronal uptake and long
neuronal retention times of .sup.11C-labeled
fluoro-phenoxyethylguanidines in the isolated rat heart model.
[0054] FIG. 14 is a plot showing the kinetics of .sup.11C-guanoxan
(GOX) and two ring-hydroxylated analogs,
.sup.11C-7-hydroxy-guanoxan (7H-GOX) and
.sup.11C-6-hydroxy-guanoxan (6H-GOX).
[0055] It is to be understood that the figures are not necessarily
drawn to scale, nor are the objects in the figures necessarily
drawn to scale in relationship to one another. The figures are
depictions that are intended to bring clarity and understanding to
various embodiments of apparatuses, systems, and methods disclosed
herein. Wherever possible, the same reference numbers will be used
throughout the drawings to refer to the same or like parts.
Moreover, it should be appreciated that the drawings are not
intended to limit the scope of the present teachings in any
way.
DETAILED DESCRIPTION
[0056] Provided herein is technology related to the preparation of
.sup.18F-labeled phenethylguanidines. In some embodiments, the
methods employ a .sup.18F-labeling step followed by one or two
simple steps to yield the final radiolabeled product. It is
contemplated that these methods permit automation of the process
and thus allow for the routine commercial preparation of the target
radiophamaceuticals at central distribution facilities. The
radiosynthetic methods provided here (see, e.g., FIGS. 1 & 2)
utilize a diaryliodonium salt precursor as a means of incorporating
fluorine-18 into the phenyl ring of a phenethylguanidine structure.
Embodiments of the methods differ in the specific structures of the
side chains of the precursors and of the .sup.18F-labeled
intermediate compounds that are ultimately converted into the
target .sup.18F-phenethylguanidine.
[0057] In the description of the technology, the section headings
used herein are for organizational purposes only and are not to be
construed as limiting the described subject matter in any way.
[0058] In this detailed description of the various embodiments, for
purposes of explanation, numerous specific details are set forth to
provide a thorough understanding of the embodiments disclosed. One
skilled in the art will appreciate, however, that these various
embodiments may be practiced with or without these specific
details. In other instances, structures and devices are shown in
block diagram form. Furthermore, one skilled in the art can readily
appreciate that the specific sequences in which methods are
presented and performed are illustrative and it is contemplated
that the sequences can be varied and still remain within the spirit
and scope of the various embodiments disclosed herein.
[0059] All literature and similar materials cited in this
application, including but not limited to, patents, patent
applications, articles, books, treatises, and internet web pages
are expressly incorporated by reference in their entirety for any
purpose. Unless defined otherwise, all technical and scientific
terms used herein have the same meaning as is commonly understood
by one of ordinary skill in the art to which the various
embodiments described herein belongs. When definitions of terms in
incorporated references appear to differ from the definitions
provided in the present teachings, the definition provided in the
present teachings shall control.
DEFINITIONS
[0060] To facilitate an understanding of the present technology, a
number of terms and phrases are defined below. Additional
definitions are set forth throughout the detailed description.
[0061] Throughout the specification and claims, the following terms
take the meanings explicitly associated herein, unless the context
clearly dictates otherwise. The phrase "in one embodiment" as used
herein does not necessarily refer to the same embodiment, though it
may. Furthermore, the phrase "in another embodiment" as used herein
does not necessarily refer to a different embodiment, although it
may. Thus, as described below, various embodiments of the
technology may be readily combined, without departing from the
scope or spirit of the technology.
[0062] In addition, as used herein, the term "or" is an inclusive
"or" operator and is equivalent to the term "and/or" unless the
context clearly dictates otherwise. The term "based on" is not
exclusive and allows for being based on additional factors not
described, unless the context clearly dictates otherwise. In
addition, throughout the specification, the meaning of "a", "an",
and "the" include plural references. The meaning of "in" includes
"in" and "on."
[0063] As used herein the term, "in vitro" refers to an artificial
environment and to processes or reactions that occur within an
artificial environment. In vitro environments may include, but are
not limited to, test tubes and cell cultures. The term "in vivo"
refers to the natural environment (e.g., an animal or a cell) and
to processes or reactions that occur within a natural
environment.
[0064] As used herein, the terms "subject" and "patient" refer to
any animal, such as a mammal like a dog, cat, bird, livestock, and
preferably a human (e.g., a human with a disease such as obesity,
diabetes, or insulin resistance).
[0065] As used herein, the term "effective amount" refers to the
amount of a composition sufficient to effect beneficial or desired
results. An effective amount can be administered in one or more
administrations, applications, or dosages and is not intended to be
limited to a particular formulation or administration route.
[0066] As used herein, the term "administration" refers to the act
of giving a drug, prodrug, or other agent, or therapeutic treatment
to a subject. Exemplary routes of administration to the human body
can be through the eyes (ophthalmic), mouth (oral), skin
(transdermal, topical), nose (nasal), lungs (inhalant), oral mucosa
(buccal), ear, by injection (e.g., intravenously, subcutaneously,
intratumorally, intraperitoneally, etc.), and the like.
[0067] As used herein, the term "co-administration" refers to the
administration of at least two agents or therapies to a subject. In
some embodiments, the co-administration of two or more agents or
therapies is concurrent. In other embodiments, a first
agent/therapy is administered prior to a second agent/therapy.
Those of skill in the art understand that the formulations and/or
routes of administration of the various agents or therapies used
may vary. The appropriate dosage for co-administration can be
readily determined by one skilled in the art. In some embodiments,
when agents or therapies are co-administered, the respective agents
or therapies are administered at lower dosages than appropriate for
their administration alone. Thus, co-administration is especially
desirable in embodiments where the co-administration of the agents
or therapies lowers the requisite dosage of a potentially harmful
(e.g., toxic) agent.
[0068] As used herein, the term "pharmaceutical composition" refers
to the combination of an active agent with a carrier, inert or
active, making the composition especially suitable for therapeutic
use.
[0069] The terms "pharmaceutically acceptable" or
"pharmacologically acceptable", as used herein, refer to
compositions that do not substantially produce adverse reactions,
e.g., toxic, allergic, or immunological reactions, when
administered to a subject.
[0070] As used herein, the term "sample" is used in its broadest
sense. In one sense, it is meant to include a specimen or culture
obtained from any source, as well as biological and environmental
samples. Biological samples may be obtained from animals (including
humans) and encompass fluids, solids, tissues, and gases.
Biological samples include blood products, such as plasma, serum
and the like. Environmental samples include environmental material
such as surface matter, soil, water, crystals and industrial
samples. Such examples are not however to be construed as limiting
the sample types applicable to the present technology.
[0071] As used herein, the terms "alkyl" and the prefix "alk-" are
inclusive of both straight chain and branched chain saturated or
unsaturated groups, and of cyclic groups, e.g., cycloalkyl and
cycloalkenyl groups. Unless otherwise specified, acyclic alkyl
groups are from 1 to 6 carbons. Cyclic groups can be monocyclic or
polycyclic and preferably have from 3 to 8 ring carbon atoms.
Exemplary cyclic groups include cyclopropyl, cyclopentyl,
cyclohexyl, and adamantyl groups. Alkyl groups may be substituted
with one or more substituents or unsubstituted. Exemplary
substituents include alkoxy, aryloxy, sulfhydryl, alkylthio,
arylthio, halogen, alkylsilyl, hydroxyl, fluoroalkyl,
perfluoroalkyl, amino, aminoalkyl, disubstituted amino, quaternary
amino, hydroxyalkyl, carboxyalkyl, and carboxyl groups. When the
prefix "alk" is used, the number of carbons contained in the alkyl
chain is given by the range that directly precedes this term, with
the number of carbons contained in the remainder of the group that
includes this prefix defined elsewhere herein. For example, the
term "C.sub.1-C.sub.4 alkaryl" exemplifies an aryl group of from 6
to 18 carbons (e.g., see below) attached to an alkyl group of from
1 to 4 carbons.
[0072] As used herein, the term "aryl" refers to a carbocyclic
aromatic ring or ring system. Unless otherwise specified, aryl
groups are from 6 to 18 carbons. Examples of aryl groups include
phenyl, naphthyl, biphenyl, fluorenyl, and indenyl groups.
[0073] As used herein, the term "heteroaryl" refers to an aromatic
ring or ring system that contains at least one ring heteroatom
(e.g., O, S, Se, N, or P). Unless otherwise specified, heteroaryl
groups are from 1 to 9 carbons. Heteroaryl groups include furanyl,
thienyl, pyrrolyl, imidazolyl, pyrazolyl, oxazolyl, isoxazolyl,
thiazolyl, isothiazolyl, triazolyl, tetrazolyl, oxadiazolyl,
oxatriazolyl, pyridyl, pyridazyl, pyrimidyl, pyrazyl, triazyl,
benzofuranyl, isobenzofuranyl, benzothienyl, indole, indazolyl,
indolizinyl, benzisoxazolyl, quinolinyl, isoquinolinyl, cinnolinyl,
quinazolinyl, naphthyridinyl, phthalazinyl, phenanthrolinyl,
purinyl, and carbazolyl groups.
[0074] As used herein, the term "heterocycle" refers to a
non-aromatic ring or ring system that contains at least one ring
heteroatom (e.g., O, S, Se, N, or P). Unless otherwise specified,
heterocyclic groups are from 2 to 9 carbons. Heterocyclic groups
include, for example, dihydropyrrolyl, tetrahydropyrrolyl,
piperazinyl, pyranyl, dihydropyranyl, tetrahydropyranyl,
dihydrofuranyl, tetrahydrofuranyl, dihydrothiophene,
tetrahydrothiophene, and morpholinyl groups.
[0075] Aryl, heteroaryl, or heterocyclic groups may be
unsubstituted or substituted by one or more substituents selected
from the group consisting of C.sub.1-6 alkyl, hydroxy, halo, nitro,
C.sub.1-6 alkoxy, C.sub.1-6 alkylthio, trifluoromethyl, C.sub.1-6
acyl, arylcarbonyl, heteroarylcarbonyl, nitrile, C.sub.1-6
alkoxycarbonyl, alkaryl (where the alkyl group has from 1 to 4
carbon atoms), and alkheteroaryl (where the alkyl group has from 1
to 4 carbon atoms).
[0076] As used herein, the term "alkoxy" refers to a chemical
substituent of the formula OR, where R is an alkyl group. By
"aryloxy" is meant a chemical substituent of the formula OR', where
R' is an aryl group.
[0077] As used herein, the term "C.sub.x-y alkaryl" refers to a
chemical substituent of formula RR', where R is an alkyl group of x
to y carbons and R' is an aryl group as defined elsewhere
herein.
[0078] As used herein, the term "C.sub.x-y alkheteroaryl" refers to
a chemical substituent of formula RR'', where R is an alkyl group
of x to y carbons and R'' is a heteroaryl group as defined
elsewhere herein.
[0079] As used herein, the term "halide" or "halogen" or "halo"
refers to bromine, chlorine, iodine, or fluorine.
[0080] As used herein, the term "non-vicinal O, S, or N" refers to
an oxygen, sulfur, or nitrogen heteroatom substituent in a linkage,
where the heteroatom substituent does not form a bond to a
saturated carbon that is bonded to another heteroatom.
[0081] As used herein, the group "R.sub.n--I" or "R--I" represents
a group wherein the iodine atom ("I") is bonded to the main
structure, unless specified otherwise. The group "X.sub.3Sn"
represents a group wherein the tin atom ("Sn") is bonded to the
main structure, unless specified otherwise.
[0082] As used herein, "Koser's reagent" refers to
hydroxy(tosyloxy)iodobenzene ("HTIB"; PhI(OTs)OH)), e.g., as
described in Koser, et al. (1982) J. Org. Chem. 47: 2487.
[0083] For structural representations where the chirality of a
carbon has been left unspecified it is to be presumed by one
skilled in the art that either chiral form of that stereocenter is
possible.
Embodiments of the Technology
[0084] The present technology provides novel compounds and novel
methods for producing compounds that find use as imaging agents
within nuclear medicine applications (e.g., PET imaging and SPECT
imaging). The present technology also provides methods for
producing imaging compositions for use within nuclear medicine
applications. Exemplary compounds and methods of the present
technology are described in more detail in the following
sections.
I. Radiotracing Agents
[0085] Nuclear Radiology is a sub-specialty of Radiology in which
radiotracing agents (e.g., compounds containing radioactive forms
of atoms) are introduced into the body for the purpose of imaging,
evaluating organ function, or localizing disease or tumors.
Radiolabelled compounds are used, for example, for both tumor
detection and tumor therapy. Many tumor cells have a higher density
of cell receptors for various circulating compounds than do
non-tumor cells; e.g., endocrine tumors show a high density of cell
surface receptors for somatostatin and brain gliomas show a high
density of receptors for epidermal growth factor. Thus, a
radiolabeled compound that binds to these cellular receptors
preferentially binds to the tumor cells. Additionally,
angiogenesis, the formation of new blood vessels from established
microvasculature, is a critical process for tumor growth. Primary
tumors and metastases will not grow beyond 2 mm in diameter without
an enhanced vascular supply. Angiogenic cells also have a higher
density of cell receptors for various circulating compounds than do
non-angiogenic vascular tissue; e.g., receptors for both
somatostatin and vascular endothelial growth factor are higher in
angiogenic tissue. Thus, a tumor can also be detected by
radiolabeled compounds binding to the angiogenic cells that are
closely associated with the tumor cells.
[0086] The present technology provides new compounds and new
methods of producing compounds useful as radiotracing agents. In
preferred embodiments, the compounds are structurally related to
meta-iodobenzylguanidine (MIBG) and possess kinetic properties
superior to MIBG for nuclear medicine applications. In particular,
the radiotracing agents of the present technology provide a slower
cellular uptake rate and a longer cellular retention length. In
preferred embodiments, the present technology provides radiolabeled
phenethylguanidines, arylalkylguanidines, aryl-Y-alkylguanidines,
and heteroarylalkylguanidines. These compounds can be radiolabeled
with several radioisotopes, including, but not limited to,
radio-halogens such as iodine-123 (.sup.123I) for single photon
imaging (e.g., SPECT imaging), iodine-131 (.sup.131I) for
radiotherapy of adrenergic tumors, and carbon-11 (.sup.11C) or
fluorine-18 (.sup.18F) for positron emission tomography imaging
(e.g., PET imaging). The technology is particularly applicable to
.sup.18F compounds.
[0087] Phenethylguanidines differ from benzylguanidines in that
they have an additional carbon atom in the side chain of the
molecule. The two-carbon side chain structure of
phenethylguanidines is similar to that of norepinephrine (NE), the
endogenous neurotransmitter of sympathetic neurons in the
heart:
##STR00032##
Additional exemplary compounds related to the technology include,
but are not limited to, (-)-beta-hydroxyphenethylguanidine,
para-methoxy-phenethylguanidine, meta-hydroxyphenethylguanidine,
para-hydroxyphenethylguanidine, 3,4-dihydroxyphenethylguanidine,
"N-guanyl-meta-octopamine", "N-guanyl-norepinephrine",
"N-guanyl-(-)-metaraminol", meta-fluorophenethylguanidine,
para-fluorophenethylguanidine, ortho-fluorophenethylguanidine,
para-fluoro-meta-hydroxy-phenethylguanidine,
ortho-fluoro-meta-hydroxy-phenethylguanidine,
meta-iodophenethylguanidine, and
para-hydroxy-meta-iodo-phenethylguanidine. In preferred
embodiments, the compounds of the present technology are
radio-labeled (e.g., .sup.11C, .sup.14C, .sup.18F, .sup.131I and
.sup.123I).
[0088] In preferred embodiments, the compounds of the present
technology are described by the following chemical formula:
##STR00033##
wherein R.sub.1, R.sub.2, R.sub.3, R.sub.4 and R.sub.5 are the same
or different and are independently selected from the group
consisting of H, halogen, hydroxyl, guanyl, methoxy, methyl, amino,
and nitro, wherein R.sub.6 is selected from the group consisting of
H and hydroxyl, and wherein R.sub.7 is H or CH.sub.3. In preferred
embodiments, the compound is selected from the group consisting of
[.sup.11C](-)-beta-hydroxyphenethylguanidine,
[.sup.11C]para-methoxy-phenethylguanidine,
[.sup.11C]meta-hydroxyphenethylguanidine,
[.sup.11C]para-hydroxyphenethylguanidine,
[.sup.11C]3,4-dihydroxyphenethylguanidine,
"N-[.sup.11C]guanyl-meta-octopamine",
"N-[.sup.11C]guanyl-norepinephrine",
"N-[.sup.11C]guanyl-(-)-metaraminol",
[.sup.11C]meta-fluorophenethylguanidine,
[.sup.11C]para-fluorophenethylguanidine, [.sup.11C]ortho
fluorophenethylguanidine,
[.sup.11C]para-fluoro-meta-hydroxy-phenethylguanidine,
[.sup.11C]ortho-fluoro-meta-hydroxy-phenethylguanidine,
[.sup.11C]meta iodophenethylguanidine, and
[.sup.11C]para-hydroxy-meta-iodo-phenethylguanidine. In preferred
embodiments, the halogen is selected from the group consisting of
.sup.18F, .sup.211At, .sup.76Br, .sup.131I, and .sup.123I.
[0089] Additional exemplary embodiments include, but are not
limited to, methods of producing compounds such as the
following:
(1) [.sup.18F], [.sup.76Br], [.sup.211At], [.sup.131I] or
[.sup.123I]-Phenethylguanidines
##STR00034##
[0090] (2) [.sup.18F], [.sup.76Br], [.sup.211At], [.sup.131I] or
[.sup.123I]-3-Hydroxyphenethylguanidines
##STR00035##
[0091] (3) [.sup.18F] or
[.sup.76Br]-3,4-Dihydroxyphenethylguanidines
##STR00036##
[0092] (4) [.sup.18F], [.sup.76Br], [.sup.211At], [.sup.131I] or
[.sup.123I]-.beta.-Hydroxyphenethylguanidines
##STR00037##
[0093] (5) [.sup.18F], [.sup.76Br], [.sup.211At], [.sup.131I] or
[.sup.123I]-3-Hydroxy-.beta.-hydroxyphenethylguanidines
##STR00038##
[0095] In some embodiments, the invention includes methods and
compounds related to arylalkylguanidines, aryl-Y-alkylguanidines,
and heteroarylalkylguanidines. Arylalkylguanidines are generally
described by the following formula:
##STR00039##
[0096] In some embodiments, .sup.18F is added to an
arylalkylguanidine compound. One compound resulting from such
fluorinations is as follows:
##STR00040##
Aryl-Y-alkylguanidines are generally described by the following
formula:
##STR00041##
[0097] In some embodiments, .sup.18F is added to an
aryl-Y-alkylguanidine compound. One compound resulting from such
fluorinations is as follows:
##STR00042##
Heteroarylalkylguanidines are generally described by the following
formulae:
##STR00043##
[0098] In some embodiments, .sup.18F is added to an
heteroarylalkylguanidine compound. Exemplar compounds resulting
from such fluorinations are as follows:
##STR00044##
II. Uses of Radiotracing Agents
[0099] The radiotracing agents of the present technology find many
uses. In particular, the radiotracing agents of the present
technology find use as imaging agents within nuclear medicine
imaging protocols (e.g., PET imaging, SPECT imaging).
[0100] In preferred embodiments, the radiotracing agents of the
present technology are useful as imaging agents within PET imaging
studies. PET is the study and visualization of human physiology by
electronic detection of short-lived positron emitting
radiopharmaceuticals. It is a non-invasive technology that
quantitatively measures metabolic, biochemical, and functional
activity in living tissue.
[0101] The PET scan is a vital method of measuring body function
and guiding disease treatment. It assesses changes in the function,
circulation, and metabolism of body organs. Unlike MRI (Magnetic
Resonance Imaging) or CT (Computed Tomography) scans that primarily
provide images of organ anatomy, PET measures chemical changes that
occur before visible signs of disease are present on CT and MRI
images.
[0102] PET visualizes behaviors of trace substances within a
subject (e.g., a living body) having a radioimaging agent
administered therein by detecting a pair of photons occurring as an
electron/positron annihilation pair and moving in directions
opposite from each other (see, e.g., U.S. Pat. No. 6,674,083,
herein incorporated by reference in its entirety). A PET apparatus
is equipped with a detecting unit having a number of small-size
photon detectors arranged about a measurement space in which the
subject is placed. The detecting unit detects frequencies of the
generation of photon pairs in the measurement space on the basis of
the stored number of coincidence-counting information items, or
projection data, and then stores photon pairs occurring as
electron/positron annihilation pairs by coincidence counting and
reconstructs an image indicative of spatial distributions. The PET
apparatus plays an important role in the field of nuclear medicine
and the like, whereby biological functions and higher-order
functions of brains can be studied by using it. Such PET
apparatuses can be roughly classified into two-dimensional PET
apparatuses, three-dimensional PET apparatuses, and
slice-septa-retractable type three-dimensional PET apparatuses.
[0103] In general, a PET detector or camera typically consists of a
polygonal or circular ring of radiation detection sensors placed
around a patient area (see, e.g., U.S. Pat. No. 6,822,240, herein
incorporated by reference in its entirety). Radiation detection
begins by injecting isotopes with short half-lives into a patient's
body placed within the patient area. The isotopes are absorbed by
target areas within the body and emit positrons. In the human body,
the positrons annihilate with electrons. As a result thereof, two
essentially monoenergetic gamma rays are emitted simultaneously in
opposite directions. In most cases the emitted gamma rays leave the
body and strike the ring of radiation detectors.
[0104] The ring of detectors includes typically an inner ring of
scintillation crystals and an outer ring of light detectors, e.g.,
photomultiplier tubes. The scintillation crystals respond to the
incidence of gamma rays by emitting a flash of light (photon
energy), so-called scintillation light, which is then converted
into electronic signals by a corresponding adjacent photomultiplier
tube. A computer, or similar, records the location of each light
flash and then plots the source of radiation within the patient's
body by comparing flashes and looking for pairs of flashes that
arise simultaneously and from the same positron-electron
annihilation point. The recorded data is subsequently translated
into a PET image. A PET monitor displays the concentration of
isotopes in various colors indicating level of activity. The
resulting PET image then indicates a view of neoplasms or tumors
existing in the patient's body.
[0105] Such detector arrangement is known to have a good energy
resolution, but relatively bad spatial and temporal resolutions.
Early PET detectors required a single photomultiplier tube to be
coupled to each single scintillation crystal, while today, PET
detectors allow a single photodetector to serve several crystals,
see e.g. U.S. Pat. Nos. 4,864,138; 5,451,789; and 5,453,623, each
herein incorporated by reference in their entireties). In such
manner the spatial resolution is improved or the number of
photodetectors needed may be reduced.
[0106] Single Photon Emission Computed Tomography (SPECT) is a
tomographic nuclear imaging technique producing cross-sectional
images from gamma ray emitting radiopharmaceuticals (single photon
emitters or positron emitters). SPECT data are acquired according
to the original concept used in tomographic imaging: multiple views
of the body part to be imaged are acquired by rotating the camera
detector head(s) (e.g., of an Anger camera) around a craniocaudal
axis. Using backprojection, cross-sectional images are then
computed with the axial field of view (FOV) determined by the axial
field of view of the gamma camera. SPECT cameras are either
standard gamma cameras that can rotate around the patient's axis or
consist of two or even three camera heads to shorten acquisition
time. Data acquisition is over at least half a circle (180.degree.)
(used by some for heart imaging), but usually over a full circle.
Data reconstruction takes into account the fact that the emitted
rays are also attenuated within the patient, e.g., photons
emanating from deep inside the patient are considerably attenuated
by surrounding tissues. While in CT, absorption is the essence of
the imaging process, in SPECT, attenuation degrades the images.
Thus, data of the head reconstructed without attenuation correction
may show substantial artificial enhancement of the peripheral brain
structures relative to the deep ones. The simplest way to deal with
this problem is to filter the data before reconstruction. A more
elegant but elaborate method used in triple head cameras is to
introduce a gamma-ray line source between two camera heads, which
are detected by the opposing camera head after being partly
absorbed by the patient. This camera head then yields transmission
data while the other two collect emission data. Note that the
camera collecting transmission data has to be fitted with a
converging collimator to admit the appropriate gamma rays.
[0107] SPECT is routinely used in clinical studies. For example,
SPECT is usually performed with a gamma camera comprising a
collimator fixed on a gamma detector that traces a revolution orbit
around the patient's body. The gamma rays, emitted by a radioactive
tracer accumulated in certain tissues or organs of the patient's
body, are sorted by the collimator and recorded by the gamma
detector under various angles around the body. From the acquired
planar images, the distribution of the activity inside the
patient's body is computed using certain reconstruction algorithms.
Generally, the so-called Expectation-Maximization of the
Maximum-Likelihood (EM-ML) algorithm is used, as described by Shepp
et al. (IEEE Trans. Med. Imaging 1982; 2:113-122) and by Lange et
al. (J. Comput. Assist. Tomogr. 1984; 8:306-316). This iterative
algorithm minimizes the effect of noise in SPECT images.
[0108] In preferred embodiments, the radiotracing agents of the
present technology are used as imaging agents for PET imaging and
SPECT imaging. It is contemplated that the radiotracing agents of
the present technology are provided to a nuclear pharmacist or a
clinician in kit form.
[0109] A pharmaceutical composition produced according to the
present technology comprises use of one of the aforementioned
radiotracing agents and a carrier such as a physiological buffered
saline solution or a physiologically buffered sodium acetate
carrier. It is contemplated that the composition will be
systemically administered to the patient as by intravenous
injection. Suitable dosages for use as a diagnostic imaging agent
are, for example, from about 0.2 to about 2.0 mCi of I-131 labeled
radiotracing agent for the adrenal medulla or tumors therein, and
from about 2.0 to about 10.0 mCi of the I-123 labeled agent for
imaging of the heart and adrenal medulla or tumors therein. For use
as a therapeutic agent, a higher dosage is required, for example,
from about 100 to about 300 mCi of the radiotracing agent
material.
[0110] It will be appreciated by those skilled in the art that the
imaging agents of the present technology are employed in accordance
with conventional methodology in nuclear medicine in a manner
analogous to that of the aforementioned radiotracing agents. Thus,
a composition of the present technology is typically systemically
applied to the patient, and subsequently the uptake of the
composition in the selected organ is measured and an image formed,
for example, by means of a conventional gamma camera.
[0111] Further understanding of use of the present technology can
be obtained from the following examples and from Kline, et al.:
"Myocardial Imaging in Man with [123 I]-Meta-Iodobenzylguanidine,"
J. Nucl. Med. 22:129-132, 1981; Wieland, et al: "Myocardial Imaging
with a Radioiodinated Norepinephrine Storage Analog," J. Nucl. Med.
22:22-31, 1981; Valk, et al: "Spectrum of Pheochromocytoma in
Multiple Endocrine Neoplasia: A Scintigraphic Portrayal Using
.sup.131I-Meta-Iodobenzylguanidine," Ann. Intern. Med., Vol. 94,
pp. 762-767 (1981); Sisson, et al.: "Scintigraphic Localization of
Pheochromocytoma," New Eng. J. Med., Vol. 305, pp. 12-17, (1981);
and Lynn, et al., "Portrayal of Pheochromocytoma and Normal Human
Adrenal Medulla by m-[I-123]-iodobenzylguanidine", J. Nucl. Med.,
Vol. 25, Vol. 436-440 (1984); and U.S. Pat. Nos. 4,584,187 and
4,622,217; these articles are specifically incorporated by
reference herein.
III. Methods of Producing 18-F Radiolabeled Phenethylguanidines
[0112] To prepare some embodiments of compounds according to the
technology described herein, a radiosynthetic scheme was used for
preparing [.sup.18F]4F-MHPG in which an intermediate fluorine-18
labeled compound 4-[.sup.18F]fluoro-meta-tyramine ([.sup.18F]4F-MT)
was produced using a conventional method for preparing
6-[.sup.18F]fluoro-dopamine (see, e.g., Ding et al. (1991)
"Synthesis of high specific activity 6-[.sup.18F]fluorodopamine for
PET studies of sympathetic nervous tissue", J Med Chem. 34: 861-3)
as modified by Langer (see, e.g., Langer et al. (2000) "High
specific radioactivity (1R,2S)-4-[.sup.18F]fluorometaraminol: a PET
radiotracer for mapping sympathetic nerves of the heart", Nucl Med
Biol. 27: 233-8; Langer et al. (2001) "Synthesis of
high-specific-radioactivity 4- and
6-[.sup.18F]fluorometaraminol-PET tracers for the adrenergic
nervous system of the heart", Bioorg Med Chem. 9: 677-94). Then,
purified [.sup.18F]4F-MT was reacted with cyanogen bromide (CNBr)
for 3 minutes at 120.degree. C. to prepare the cyanamide
intermediate. This was then reacted with NH.sub.4Br/NH.sub.4OH for
15 minutes at 130.degree. C., followed by HPLC purification, to
yield the target compound [.sup.18F]4F-MHPG. The radiosynthesis was
reproducible and the final product was routinely prepared at
>95% radiochemical purity. Some steps of this synthesis were
performed manually; in addition, the synthesis took approximately 4
hours to complete and the synthesis provided relatively low
radiochemical yields and low specific activities. See, e.g., Jang
et al (2013) "Synthesis and bioevaluation of
[.sup.18F]4-fluoro-m-hydroxyphenethylguanidine ([.sup.18F]4F-MHPG):
a novel radiotracer for quantitative PET studies of cardiac
sympathetic innervation", Bioorg Med Chem Lett. 23: 1612-6.
[0113] Conventional reaction schemes, or reaction schemes
comprising one or more conventional steps, for producing compounds
according to the technology (e.g., a .sup.18F-labeled
phenethylguanidine) were tested and found to be unsatisfactory for
automated synthesis, e.g., due to high pressures developed in the
reaction vial when heating with NH.sub.4Br/NH.sub.4OH at
130.degree. C., due to the use of LiAlH.sub.4, steps requiring
manual steps performed by hand, and/or requiring long reaction
and/or work-up times. Thus, alternative approaches were developed
for an automated radiosynthesis.
[0114] For example, in one embodiment, the methods related to the
technology involve using a diaryliodonium salt precursor in which
the entire side chain, including the guanidine moiety itself, is
unprotected (R.sub.8, R.sub.9, R.sub.10, and/or R.sub.11=hydrogen)
or protected (one or more of R.sub.8, R.sub.9, R.sub.10, and/or
R.sub.11=N-protecting group), directly yielding a .sup.18F-labeled
phenethylguanidine structure (see FIG. 1). During the development
of this technology, a model compound (R.sub.3=R.sub.12--I, R.sub.1,
R.sub.2, R.sub.4-R.sub.11=hydrogen) was tested and found to produce
a 7% radiochemical yield in the .sup.18F-labeling step. Similarly,
a second model compound (R.sub.3=R.sub.12--I, R.sub.1, R.sub.2,
R.sub.4-R.sub.7=hydrogen, R.sub.8-R.sub.11=tert-Boc) was tested.
After .sup.18F-labeling and treatment with mild acid for simple
N-Boc deprotection, followed by HPLC purification, this second
model compound was found to produce the final target product
(4[.sup.18F]fluoro-phenethylguanidine) at 10% radiochemical yield
(end of synthesis).
[0115] In another embodiment, the methods relate to a two-step
reaction to prepare a .sup.18F-phenethylguanidine (see FIG. 2A).
The .sup.18F-labeling step uses a diaryliodonium salt precursor
with an N-protected phenethylamine moiety instead of the guanidine
group used in the first method. The
N-Boc-aminoethylphenyl(2-thienyl) iodonium salt provides very high
radiochemical yields in the .sup.18F-labeling step. Treatment with
mild acid for simple N-Boc deprotection followed by purification
using a C18 Sep-Pak delivers 4-[.sup.18F]fluoro-meta-tyramine as an
intermediate. This is then reacted with a guanylating agent
(N,N'-diBoc-5-chlorobenzotriazole) to convert the primary amine
into a guanidine. Cleavage of both of the benzyloxy and N-Boc
protecting groups with acid in the final step generates the
.sup.18F-labeled phenethylguanidine, e.g.,
4-[.sup.18F]fluoro-meta-hydroxy
phenethylguanidine([.sup.18F]4F-MHPG).
[0116] In yet another embodiment, an automated method was developed
for the preparation of [.sup.18F]4F-MHPG (see, e.g., FIG. 2B). In
this embodiment, a single .sup.18F-labeling step is followed by
additional steps to yield the final product. In some embodiments,
two reaction modules (e.g., GE TRACERlab FX.sub.FN modules) (in
adjacent hot-cells) were used in sequence for a fully automated
synthesis of the [.sup.18F] compound 1 (FIG. 2B). The first
FX.sub.FN module was used for production of
3-benzyloxy-4-[.sup.18F]fluoro-meta-tyramine [.sup.18F](FIG. 2B,
compound 4), while the second FX.sub.FN module was used to convert
the [.sup.18F] compound 4 (FIG. 2B) into the final product 1, as
shown in FIG. 2B. In the first step of the reaction, the iodonium
salt precursor (FIG. 2B, compound 2) was reacted with Cs[.sup.18F]F
in DMF containing the radical scavenger TEMPO to prepare the
[.sup.18F] compound 3 (FIG. 2B). Removal of the Boc protecting
group from 3 using 1.0 N HCl provided the intermediate [.sup.18F]
compound 4. In general, approximately 1.4 Ci of [.sup.18F]F- was
used and 170-250 mCi of [.sup.18F] compound 4 was obtained with a
15.+-.3% radiochemical yield and >95% radiochemical purity.
Compound 4 was purified using a C-18 Sep-Pak cartridge and eluted
into the reaction vial of the second FX.sub.FN module.
[0117] The transferred solution comprising [.sup.18F] compound 4
(170-250 mCi) was evaporated under a stream of nitrogen and then
cooled. A solution of
N,N'-bis-(tert-butoxycarbonyl)-5-chloro-1H-benzotriazole-1-carboxamidine
(FIG. 2B, compound 5) in a mixed solution of DIEA and MeCN was
added to the reactor vessel containing [.sup.18F] compound 4 and
the resulting mixture was heated at 45.degree. C. for 15 minutes to
form [.sup.18F] compound 6 (FIG. 2B). Next, the simultaneous
removal of the benzylether protecting group and the N,N'-di-Boc
protecting groups was achieved by adding 1.0 N HBr to the reactor
vessel and heating at 120.degree. C. for 15 minutes. After cooling,
a mixture of 1.0 N NaOH and a buffer solution was added to the
reaction vial. The crude product was injected onto a reverse-phase
HPLC column (Phenomenex Synergi 10 micron Hydro-RP 80A,
250.times.10 mm, 5% EtOH in 60 mM NaH.sub.2PO.sub.4 buffer, flow
rate 4.0 mL/min, .lamda..sub.224 nm) and [.sup.18F] compound 1
(FIG. 2B) was collected, e.g., at approximately 28-30 minutes.
Typically, 55-125 mCi of [.sup.18F] compound 1 was obtained with a
7.+-.3% overall radiochemical yield and >99% radiochemical
purity. The specific activity (SA) averaged 1.2.+-.0.3 Ci/.mu.mol.
Total synthesis time from end of bombardment (EOB) was
approximately 150 minutes. Some embodiments contemplate similar
syntheses on a synthesizer having two reaction vials (e.g., a GE
TRACERlab FX N Pro Synthesizer) to perform the synthesis in a
single automated radiosynthesis module.
[0118] It is contemplated that these methods are used in certain
embodiments to produce other .sup.18F-phenethylguanidine structures
such as those provided in U.S. Pat. No. 7,534,418, which is
incorporated herein by reference in its entirety for all
purposes.
[0119] Moreover, based on the simplicity and robust yields of this
method, it is contemplated that this second approach is automated
in particular embodiments to produce sufficiently large batches of
[.sup.18F]4F-MHPG at high specific activities and high
radiochemical yields to prepare and distribute the compounds daily
to stand-alone PET centers.
IV. Application of 18F-Labeling Method to Producing Additional
Guanidine Imaging Agents
[0120] In addition to the embodiment of the technology described
above for preparing .sup.18F-labeled phenethylguanidines, the
technology comprises embodiments in which the same approach (or
similar approaches) is used to prepare other .sup.18F-labeled
guanidine compounds that are useful as PET imaging agents. These
include, but are not limited to: [0121] A. .sup.18F-labeled
arylalkylguanidines, including, but not limited to,
.sup.18F-benzylguanidines and .sup.18F-arylpropylguanidines, such
as 4-[.sup.18F]fluoro-meta-hydroxy-benzylguanidine and
4-[.sup.18F]fluoro-meta-hydroxy-phenpropylguanidine. [0122] B.
.sup.18F-labeled aryl-Y-alkylguanidines, in which Y is O, S, or NH,
such as 4-[.sup.18F]fluoro-meta-hydroxy-phenoxyethylguanidine.
[0123] C. .sup.18F-labeled heteroarylalkylguanidines, such as
6-[.sup.18F]fluoro-7-hydroxy-guanoxan.
[0124] The .sup.18F-labeling methodology described above is used in
some embodiments to prepare .sup.18F-labeled benzylguanidines.
Embodiments of these methods are distinct from conventional
approaches for preparing compounds such as
meta-[.sup.18F]fluoro-benzylguanidine (see, e.g., Garg, et al.
(1994) "Synthesis and preliminary evaluation of para- and
meta-[.sup.18F]fluorobenzylguanidine", Nucl Med Biol 21: 97-103).
It is contemplated that novel imaging agents comprising
[.sup.18F]-labeled arylpropylguanidines are useful PET imaging
agents of cardiac sympathetic innervation and adrenergic
tumors.
[0125] The labeling technology is not limited to particular
.sup.18F-labeled arylalkylguanidines, aryl-Y-alkylguanidines,
and/or heteroarylalkylguanidines. For example, the technology
relates to embodiments of arylalkylguanidine compounds having a
general structure:
##STR00045##
embodiments of aryl-Y-alkylguanidine compounds having the
structure:
##STR00046##
and embodiments of heteroarylalkylguanidine compounds having the
structures:
##STR00047##
[0126] Provided herein are technologies related to methods of
producing and/or manufacturing .sup.18F-labeled
arylalkylguanidines, aryl-Y-alkylguanidines, and/or
heteroarylalkylguanidines, e.g., for use as imaging agents, e.g.,
in PET imaging. For example, some embodiments provide methods in
which an .sup.18F-labeled arylalkylguanidine is produced from an
iodonium salt precursor by a single step reaction in solution,
e.g.,
##STR00048##
The technology provides related embodiments in which an
arylalkylguanidine is produced from an iodonium salt precursor in a
single step using a linker, e.g.,
##STR00049##
(wherein in this and other structures X- is a counterion) and
embodiments in which an arylalkylguanidine is produced from an
iodonium salt precursor in solution by the two-step reaction:
##STR00050##
[0127] In addition, some embodiments provide methods in which an
.sup.18F-labeled aryl-Y-alkylguanidine is produced from an iodonium
salt precursor by a single step reaction in solution, e.g.,
##STR00051##
The technology provides related embodiments in which an
aryl-Y-alkylguanidine is produced from an iodonium salt precursor
in a single step using a linker, e.g.,
##STR00052##
and embodiments in which an aryl-Y-alkylguanidine is produced from
an iodonium salt precursor in solution by the two-step
reaction:
##STR00053##
[0128] Finally, some embodiments provide methods in which an
.sup.18F-labeled heteroarylalkylguanidine is produced from an
iodonium salt precursor by a single step reaction in solution,
e.g.,
##STR00054##
The technology provides related embodiments in which a
heteroarylalkylguanidine is produced from an iodonium salt
precursor in a single step using a linker, e.g.,
##STR00055##
and embodiments in which a heteroarylalkylguanidine is produced
from an iodonium salt precursor in solution by the two-step
reaction:
##STR00056##
[0129] Example 2 demonstrates that compositions comprising
.sup.18F-labeled aryl-Y-alkylguanidine compounds are successful as
PET imaging agents. These results are based on evaluating
.sup.11C-labeled analogs of phenoxyethylguanidine as potential
imaging agents for cardiac sympathetic innervation and adrenergic
tumors in an isolated rat heart system.
[0130] Heterocyclic compounds with side chains terminating with a
guanidine group are good substrates of the norepinephrine
transporter (NET) and have pharmacological activity in cardiac
sympathetic innervation. See, e.g., Broadley K J. Autonomic
Pharmacology. London: Taylor & Francis (1996). For example, the
heteroarylalkylguanidine compound
2-(2,3-dihydro-1,4-benzodioxin-2-ylmethyl) guanidine, which is
known as "guanoxan" has pharmacological activity, in particular,
guanoxan exhibits "sympatholytic" activity that prevents the
release of norepinephrine from nerve terminals. Example 3 shows
that a guanoxan compound is useful as an imaging agent. As shown in
FIG. 14, ring-hydroxylated analogs of guanoxan showed favorable
uptake and retention time. As such, these data show that an
.sup.18F-labeled analog of a ring hydroxylated analog of guanoxan,
(e.g., 6-[.sup.18F]fluoro-7-hydroxy-guanoxan) is useful as an
imaging agent, for example to image adrenergic tumors and cardiac
sympathetic innervation.
[0131] In summary, in addition to the technology related to the
.sup.18F-labeling method for preparing ring .sup.18F-fluorinated
phenethylguanidines for PET imaging, the technology provided herein
finds use to synthesize several other classes of guanidine
compounds with a ring .sup.18F-fluorine moiety. For example, as
shown by the examples and as discussed herein, several classes of
imaging agents with structural similarity to phenethylguanidines
(e.g., arylalkylguanidines, aryl-Y-alkylguanidines, and
heteroarylalkylguanidines) were evaluated and the results
demonstrate that these additional agents share a unified clinical
application with .sup.18F-labeled phenethylguanidines as imaging
agents. The specific imaging targets of these imaging agents
include, but are not limited to, the sympathetic innervation of the
heart and other organs, the adrenal medulla, and neuroendocrine
tumors such as neuroblastoma and pheochromocytoma.
[0132] Although the disclosure herein refers to certain illustrated
embodiments, it is to be understood that these embodiments are
presented by way of example and not by way of limitation.
Examples
Example 1
Biological Evaluations of [.sup.18F]4F-MHPG
[0133] During the development of the technology provided herein,
experiments were performed to evaluate high specific activity
[.sup.18F]4F-MHPG made with embodiments of the radiosynthetic
methods described herein. These results demonstrate that the
methods provide the compounds at sufficiently high specific
activities for clinical studies.
[0134] 1.1. Isolated Rat Heart Study of High Specific Activity
[.sup.18F]4F-MHPG.
[0135] Data were collected to determine the kinetics of
[.sup.18F]4F-MHPG in the isolated working rat heart model and these
data were compared with a previous study of high specific activity
[.sup.11C]4F-MHPG (FIG. 3). The radiotracer was infused into the
heart for 10 minutes to measure the tracer uptake rate into cardiac
sympathetic neurons (K.sub.up, shown in units of ml/min/g tissue
wet). Then, the heart was infused with normal heart perfusate to
measure the tracer clearance rate from the heart over 2 hours. As
shown in FIG. 3, the kinetics of the two compounds are almost
identical, independent of whether the 4F-MHPG compound is
radiolabeled with carbon-11 or fluorine-18. As such, the kinetics
of [.sup.18F]4F-MHPG are almost identical to those of
[.sup.11C]4F-MHPG in the isolated rat heart. Moreover, these
results demonstrate that [.sup.18F]4F-MHPG has a long retention
time inside sympathetic neurons (T.sub.1/2>24 hours), which
makes tracer kinetic modeling of myocardial kinetics in humans
simpler and more robust than is currently possible with existing
cardiac sympathetic nerve imaging agents.
[0136] 1.2. Biological Studies of High Specific Activity
[.sup.18F]4F-MHPG in Non-Human Primates.
[0137] During the development of embodiments of the technology,
several studies of [.sup.18F]4F-MHPG were performed in rhesus
macaque monkeys. These include assessments of image quality, in
terms of relative uptake of the radiotracer into heart, lungs and
liver; metabolic breakdown of [.sup.18F]4F-MHPG in blood;
myocardial kinetics of [.sup.18F]4F-MHPG and their quantitative
analysis; specificity of [.sup.18F]4F-MHPG for sympathetic nerve
terminals through pharmacological blocking studies; and assessment
of [.sup.18F]4F-MHPG uptake into adrenal glands (to assess its
potential application for oncology in imaging adrenergic tumors
like pheochromocytoma and neuroblastoma).
[0138] 1.2.1. MicroPET Imaging Studies of [.sup.18F]4F-MHPG.
[0139] A series of microPET imaging studies were performed in the
macaque model to assess image quality and tracer kinetics. In most
of these the time course of the metabolic breakdown of
[.sup.18F]4F-MHPG in plasma was also determined. The studies are
summarized below in Table 1.
TABLE-US-00001 TABLE 1 MicroPET studies of [.sup.18F]4F-MHPG Organ
Imaging Time Metabolism Date Conditions Imaged (min) study? Feb. 3,
2011 Control Heart 90 No Mar. 3, 2011 Control Heart 90 Yes Mar. 17,
2011 Control Heart 90 Yes Mar. 31, 2011 Control Heart 90 Yes Apr.
7, 2011 Control Heart 90 Yes May 5, 2011 Control Adrenal 90 No
glands Jun. 9, 2011 Desipramine Heart & 90 Yes (DMI) block
Adrenals of NET
[0140] Orthogonal slice images of cardiac sympathetic innervation
from three of the control studies were acquired. The high quality
images showed uniform left ventricular uptake of [.sup.18F]4F-MHPG,
consistent with uniform innervation in the left ventricle. There
was also very low uptake of the compound in the lungs and liver.
Liver uptake of [.sup.18F]4F-MHPG is much lower than is typically
seen with either [.sup.11C]meta-hydroxyephedrine (HED) or
[.sup.123I]metaiodobenzylguanidine (MIBG), providing improved image
contrast between heart tissue and nearby organs. Faint uptake in
the spinal column bones suggests there is little in vivo
defluorination of [.sup.18F]4F-MHPG.
[0141] 1.2.2 Metabolism of [.sup.18F]4F-MHPG in Monkeys.
[0142] In some of the microPET studies, venous blood samples (e.g.,
n=6) were drawn to assess the metabolic breakdown of
[.sup.18F]4F-MHPG in plasma (see Table 1). Plasma was processed for
injection onto a reverse-phase HPLC system equipped with a
radiation detector. The percentages of intact parent tracer and
radiolabeled metabolites were determined as a function of time. A
representative HPLC trace for a blood sample drawn at t=2 minutes
after injection is shown in FIG. 4A. In this system,
[.sup.18F]4-MHPG has a retention time R.sub.t of approximately 11.2
minutes while the main polar radiometabolite formed has an R.sub.t
of approximately 7.9 minutes. These data show that [.sup.18F]4-MHPG
is metabolized relatively rapidly, e.g., in the rhesus monkey.
Together, all of the data show that [.sup.18F]4F-MHPG is rapidly
metabolized in the plasma of rhesus monkeys (see, e.g., the example
shown in FIG. 5).
[0143] The main radiometabolite formed is more polar than the
parent compound, [.sup.18F]4-MHPG. The metabolite has been
identified as the sulfur conjugate of [.sup.18F]4-MHPG based on in
vitro incubations of the parent compound with a monkey liver
cytosol fraction and the required cofactor
3'-phospho-adenosine-5'phosphosulfate (PAPS). After only 1 minute
of in vitro incubation, approximately 70% of [.sup.18F]4-MHPG was
sulfur conjugated, and after 20 minutes of incubation, 100% was in
the sulfur-conjugated form (FIG. 4B). Sulfur conjugation occurs at
the meta-hydroxyl group (FIG. 4C). Without being bound by theory
and without requiring an understanding of the mechanism to practice
the technology, it is contemplated that rapid metabolism of
[.sup.18F]4F-MHPG is a reason for its low liver uptake. In
comparison, the structurally similar carbon-11 compound,
[.sup.11C]guanyl-meta-octopamine (GMO), has a slower metabolic
breakdown in monkey plasma, with a T.sub.1/2 for breakdown of
approximately 13 minutes.
[0144] Many tracers are metabolized more quickly in monkeys than in
humans, e.g., tracer metabolism rates are typically 2 to 3 times
slower in humans than in monkeys. In some embodiments, a longer
lifetime in human subjects is advantageous, e.g., to allow cardiac
neurons to accumulate the tracer for a longer period of time, thus
providing more kinetic data for quantitative analyses. In addition,
a metabolism rate that is 2 to 3 times slower in humans allows an
imaging study of approximately 30 minutes to provide all the
kinetic data needed for quantifying cardiac nerve density. Also,
metabolic breakdown essentially "inactivates" the tracer, which
could increase its safety in humans. Since [.sup.18F]4-MHPG may
have some activity as a depletor of norepinephrine from storage
vesicles in sympathetic neurons, it may prove beneficial to have it
completely metabolized to the sulfur conjugate within 1 hour after
injection into a human subject.
[0145] 1.2.3 Myocardial Kinetics of [.sup.18F]4F-MHPG
[0146] The kinetics of [.sup.18F]4-MHPG in monkeys are shown in
FIG. 6. In blood, after the peak of the bolus injection, blood
activity levels stay roughly constant from approximately 5-10
minutes to the end of the study (e.g., approximately 85 minutes).
Because of the relatively rapid metabolism (e.g., by sulfur
conjugation) of [.sup.18F]4-MHPG, after 10-20 minutes most of the
activity in blood is in the form of the sulfur conjugate of
[.sup.18F]4-MHPG. Myocardial activity climbs for about 15-30
minutes after injection and then remains constant. The lack of any
further accumulation of [.sup.18F]4-MHPG into cardiac nerve
terminals after this time is indirect evidence that the sulfur
conjugate of [.sup.18F]4-MHPG is inactive at the norepinephrine
transporter (NET). Thus only intact `parent` tracer molecules of
[.sup.18F]4-MHPG are capable of being taken up into presynaptic
sympathetic nerve terminals. Based on a predicted slower metabolism
of [.sup.18F]4-MHPG in humans than in monkeys, the tracer is
expected to accumulate for a longer time than is seen in
monkeys.
[0147] During the development of embodiments of the technology
provided herein, the kinetics of [.sup.18F]4F-MHPG in monkey hearts
was assessed using either compartmental modeling or Patlak
graphical analysis to obtain quantitative measures of regional
cardiac sympathetic nerve density.
[0148] For some analyses, a simplified compartmental model that
assumed irreversible trapping inside nerve terminals was used to
estimate the model rate constants K.sub.1 (ml/min/g), k.sub.2
(min.sup.-1), k.sub.3 (min.sup.-1), and a blood volume fraction BV
(dimensionless) (see, e.g., FIG. 7). In both the control studies
and DMI blocking studies, the model quickly converged to a single
global minimum in just a few iterations of the nonlinear regression
analysis (FIG. 8). The rate constant k.sub.3, which reflects the
neuronal uptake rate of the tracer, did not decline in proportion
to increasing doses of DMI. This shows that a fit of the model to
the data is provided by adjusting rate constants other than
k.sub.3. Combining the rate constants K.sub.1, k.sub.2, and k.sub.3
into a `net uptake rate constant` K.sub.i
(ml/min/g)=(K.sub.1k.sub.3)/(k.sub.2+k.sub.3) provides for control
studies that K=0.341.+-.0.041 ml/min/g. This aggregate parameter
provides a stable measure of cardiac nerve density that declined in
a dose-dependent manner with increasing DMI dose (FIG. 9, "Kinetic
modeling"). These results show that compartmental modeling of
[.sup.18F]4F-MHPG kinetics in human heart provides accurate and
sensitive quantitative measures of regional cardiac sympathetic
nerve density.
[0149] For some analyses, Patlak graphical analysis was used, which
uses a mathematical transformation of the kinetic data C.sub.p(t)
and C.sub.t(t) to generate a `Patlak plot` having a characteristic
linear phase (see, e.g., Patlak and Blasberg (1985) "Graphical
evaluation of blood-to-brain transfer constants from multiple-time
uptake data. Generalizations." J Cereb Blood Flow 5: 584-90). The
slope of the linear portion of a Patlak plot, K.sub.p (ml/min/g),
provides an alternate estimate of the `net uptake rate constant`
K.sub.i (ml/min/g). Thus, for the model structure used (FIG. 7),
the slope of the Patlak plot is given by
K.sub.p.apprxeq.(K.sub.1k.sub.3)/(k.sub.2+k.sub.3).
[0150] During the development of embodiments of the technology
provided herein, the myocardial kinetics of [.sup.18F]4F-MHPG was
assessed using Patlak graphical analysis and metabolite-corrected
plasma input functions. Patlak graphical analysis of one of the
control studies in a rhesus macaque monkey is shown in FIG. 10. In
this example, the Patlak plot is highly linear (r.sup.2=1.00) and
the estimated Patlak slope K.sub.patlak=0.252 ml/min/g. These data
show that a tracer possessing a relatively slow NET transport rate
and very efficient intraneuronal trapping inside storage vesicles
is appropriate for analysis using the Patlak graphical method. As
such, regional estimates of the Patlak slope from Patlak analysis
of the kinetics of [.sup.18F]4F-MHPG serve as robust measures of
regional cardiac sympathetic nerve density in patients with heart
diseases.
[0151] Additional data were collected and Patlak analyses
performed. In these experiments, for the control studies, the
Patlak plots were highly linear (r.sup.2>0.99) with an average
measured Patlak slope K.sub.p=0.302.+-.0.031 ml/min/g (FIG. 11).
For the four different DMI block studies, the data were best fit
with linear Patlak plots and the measured Patlak slopes declined in
a DMI-dose dependent manner (FIG. 9, "Patlak analysis"). The
decline in the estimated K.sub.i or K.sub.p values with increasing
doses of DMI each followed a sigmoidal dose-response curve (FIG.
9). For the K.sub.i data from compartmental modeling, a
half-maximal inhibitory concentration (IC.sub.50) of 0.051 mg/kg
DMI was estimated, with a Hill slope n.sub.H=-0.79. For the K.sub.p
data, IC.sub.50=0.025 mg/kg DMI and n.sub.H=-0.68.
[0152] These results suggest that estimates of the `net uptake
rate` constant K.sub.i, obtained from either compartmental modeling
or Patlak analysis of [.sup.18F]4F-MHPG kinetics, are sensitive and
robust measures of regional cardiac sympathetic nerve density. In
addition, these data demonstrate that the quantitative analysis
methods work over a wide range of nerve densities, as produced
experimentally with pharmacological blockade of norepinephrine
transporters (NET) using the drug desipramine (DMI).
[0153] 1.2.4 Desipramine (DMI) Block of the Neuronal Uptake of
[.sup.18F]4F-MHPG
[0154] To verify that presynaptic sympathetic nerve terminals are
the main locus of retention of [.sup.18F]4F-MHPG
([.sup.18F]4F-MHPG) in the heart, data were collected from microPET
imaging in a monkey that had been administered a blocking dose of
the potent NET inhibitor desipramine (DMI). Orthogonal slice images
were acquired for comparison with data from a control study. By
comparing microPET images of [.sup.18F]4F-MHPG in a control monkey
following different levels of DMI block of NET, the data show that
increased levels of DMI block of NET transporters on presynaptic
sympathetic nerve terminals led to a progressive decline in the
cardiac retention of the [.sup.18F]4F-MHPG tracer in myocardial
tissue and/or prevents any significant cardiac retention of the
[.sup.18F]4F-MHPG tracer. At the highest DMI block conditions (1.0
mg/kg), myocardial levels of [.sup.18F]4F-MHPG are lower than the
levels in blood, indicating complete blockade of the retention of
the tracer in the heart. The data showed that [.sup.18F]4F-MHPG is
100% specific for cardiac sympathetic neurons, with very low
nonspecific binding.
[0155] As such, myocardial uptake of [.sup.18F]4F-MHPG under
control conditions is 100% specific to sympathetic nerve terminals.
Also, in the DMI block images, data from the highest dose DMI block
study demonstrated that nonspecific binding in heart tissue outside
sympathetic nerve terminals is extremely low. Thus, in some
embodiments, quantitative measures based on kinetics and/or
retention of [.sup.18F]4F-MHPG provide accurate measures of nerve
density over a wide dynamic range, from normal levels down to the
very low levels seen in diseases that cause severe denervation,
such as diabetic autonomic neuropathy and Parkinson's disease. As
such, for this and other reasons the compounds are contemplated to
find use in diseases that cause severe denervation, such as
diabetic autonomic neuropathy and Parkinson's disease
[0156] 1.2.5 Oncology Applications: Uptake of [.sup.18F]4F-MHPG
into Adrenal Glands
[0157] To assess the ability of [.sup.18F]4F-MHPG to serve an
additional clinical role as a marker of adrenergic tumors such as
neuroblastoma and pheochromocytoma, data were collected to examine
the uptake of [.sup.18F]4F-MHPG into the adrenal gland. In control
studies, the adrenal glands took up sufficient [.sup.18F]4F-MHPG to
be visualized, especially on the right side of the transaxial
images, which was better separated from the kidney than the
contralateral adrenal gland. There was also significant uptake in
the cortex of the kidney. The kinetics of [.sup.18F]4F-MHPG in the
adrenal gland are very similar to those seen in the heart. In the
DMI block study above, the adrenal glands were scanned for 10
minutes following the 90 minute cardiac study. No uptake of
[.sup.18F]4F-MHPG was seen in the adrenal glands, demonstrating
that [.sup.18F]4F-MHPG uptake into the adrenal gland is primarily
mediated by NET transport.
[0158] In images comparing the uptake of [.sup.18F]4F-MHPG into the
adrenal glands of a control monkey and following DMI block of NET
uptake, controls show a high uptake into the renal cortex as well
as into the adrenal gland while the images from experiments testing
DMI block conditions showed that [.sup.18F]4F-MHPG uptake into the
adrenals is completely absent. These results indicate that adrenal
uptake of [.sup.18F]4F-MHPG is specific to NET transport.
[0159] These data demonstrate that [.sup.18F]4F-MHPG images adrenal
glands and is thus a tracer for adrenergic tumors (e.g.,
pheochromocytoma, neuroblastoma, etc.) using PET scanning
technologies. Together, these data show that this agent finds use
in oncology applications in the nuclear medicine clinic, e.g., for
use similar to the use of .sup.123I-meta-iodobenzyl-guanidine
(under the trade name `AdreView`, GE Healthcare) for oncology
applications, and now, more recently as a potential cardiac
sympathetic nerve imaging agent (see, e.g., Jacobson et al. (2010)
"Myocardial iodine-123 meta-iodobenzylguanidine imaging and cardiac
events in heart failure: results of the prospective ADMIRE-HF
(AdreView Myocardial Imaging for Risk Evaluation in Heart Failure)
study", J Am Coll Cardiol. 55: 2212-21).
[0160] 1.2.6 Biodistribution in a Mammal
[0161] During the development of embodiments of the technology,
data were collected from bioevaluation studies of
[.sup.18F]4F-MHPG. In particular, biodistribution studies in rats
showed that the tissue distribution of low specific activity
[.sup.18F]4F-MHPG was similar to the distribution of the carbon-11
labeled analog [.sup.11C]4F-MHPG (Table 2). Table 2 shows the
tissue concentrations of the compounds (expressed as a percentage
of the injected dose per gram) at 30 minutes after injection in
rats. A number of 5 rats was tested in each study (n=5).
TABLE-US-00002 TABLE 2 Biodistribution in rat left adrenal Tracer
ventricle lung liver muscle spleen medulla blood [.sup.11C]4F-MHPG
1.77 .+-. 0.27 0.56 .+-. 0.15 1.22 .+-. 0.18 0.07 .+-. 0.01 0.52
.+-. 0.10 0.56 .+-. 0.21 0.08 .+-. 0.01 [.sup.18F]4F-MHPG 1.45 .+-.
0.13 0.43 .+-. 0.08 1.14 .+-. 0.16 0.07 .+-. 0.01 0.56 .+-. 0.10
0.54 .+-. 0.08 0.07 .+-. 0.01
[0162] 1.2.7 Radiation Absorbed Dose Estimates
[0163] During the development of embodiments of the technology
provided herein, the biodistribution of [.sup.18F]4F-MHPG into the
organs of rats was assessed. In particular, four time points (Table
3) were collected and used to derive human radiation absorbed dose
estimates using the OLINDA/EXM 1.0 software package (see, e.g.,
Stabin et al. (2005) "OLINDA/EXM: the second-generation personal
computer software for internal dose assessment in nuclear
medicine", J Nucl Med. 46: 1023-7). From the acquired data, the
maximum percentage of the injected dose observed in the
gastrointestinal tract (12.6%) was assumed to enter the small
intestine using the ICRP 30 GI Tract model incorporated into the
OLINDA/EXM program. Also, in separate microPET imaging studies in
rats, 19.9.+-.0.2% of the injected dose was found to be excreted
through the bladder with a biological half-time of 0.20.+-.0.02
hours. These values were entered into the Dynamic Bladder model of
OLINDA/EXM, assuming a 4.0-hour void interval. Absorbed dose
estimates for the reference adult male organ model of OLINDA/EXM
are shown in Table 4. Organs with the highest absorbed dose
estimates included the urinary bladder wall (0.666 rad/mCi), upper
lower intestine (0.221 rad/mCi), small intestine (0.201 rad/mCi),
and the heart wall (0.201 rad/mCi). An `effective dose` of 0.091
rem/mCi was estimated for [.sup.18F]4F-MHPG. Under United States
Federal Regulations governing research with a new
radiopharmaceutical (21 CFR .sctn.361.1), the maximum allowable
radiation absorbed dose to an individual organ (other than the
gonads) is 5 rad. Considering the maximum estimated absorbed dose
(urinary bladder wall; 0.666 rad/mCi), initial PET studies in human
subjects could use a maximum of 7.5 mCi of [.sup.18F]4F-MHPG. This
would correspond to an `effective dose` of 0.68 rem.
TABLE-US-00003 TABLE 3 Biodistribution of [.sup.18F]4F-MHPG into
major organs in rats (% ID kg/g) Organ 5 min 30 min 60 min 120 min
Brain 0.004 .+-. 0.001 0.003 .+-. 0.000 0.002 .+-. 0.000 0.001 .+-.
0.001 Eyeballs 0.039 .+-. 0.006 0.040 .+-. 0.004 0.041 .+-. 0.006
0.026 .+-. 0.006 Heart 0.746 .+-. 0.067 0.602 .+-. 0.111 0.526 .+-.
0.116 0.347 .+-. 0.055 Lung 0.267 .+-. 0.045 0.136 .+-. 0.032 0.091
.+-. 0.015 0.036 .+-. 0.006 Liver 0.477 .+-. 0.110 0.406 .+-. 0.071
0.350 .+-. 0.077 0.233 .+-. 0.049 Pancreas 0.137 .+-. 0.024 0.145
.+-. 0.021 0.172 .+-. 0.042 0.121 .+-. 0.016 Spleen 0.268 .+-.
0.044 0.262 .+-. 0.055 0.253 .+-. 0.054 0.235 .+-. 0.066 Adrenal
0.203 .+-. 0.083 0.181 .+-. 0.069 0.193 .+-. 0.053 0.133 .+-. 0.021
Kidney 1.398 .+-. 0.530 0.183 .+-. 0.055 0.186 .+-. 0.090 0.078
.+-. 0.024 Stomach 0.116 .+-. 0.021 0.110 .+-. 0.004 0.115 .+-.
0.029 0.089 .+-. 0.019 Ovary* 0.194 .+-. 0.048 0.171 .+-. 0.020
0.142 .+-. 0.052 0.129 .+-. 0.029 Uterus* 0.268 .+-. 0.044 0.155
.+-. 0.001 0.112 .+-. 0.046 0.081 .+-. 0.006 Testes* 0.018 .+-.
0.002 0.010 .+-. 0.001 0.008 .+-. 0.003 0.004 .+-. 0.001
Muscle.sup..dagger. 0.019 .+-. 0.005 0.025 .+-. 0.011 0.016 .+-.
0.002 0.015 .+-. 0.002 Bone.sup..dagger. 0.060 .+-. 0.015 0.039
.+-. 0.015 0.027 .+-. 0.009 0.016 .+-. 0.005 Blood.sup..dagger.
0.062 .+-. 0.004 0.022 .+-. 0.003 0.016 .+-. 0.002 0.009 .+-. 0.001
*(n = 2) animals used for these values. .sup..dagger.Values
expressed as % ID/g.
TABLE-US-00004 TABLE 4 Radiation absorbed dose estimates (rem/mCi)
for [.sup.18F]4F-MHPG for the reference adult male model of
OLINDA/EXM 1.0 Total dose Dose for 10 mCi Target organ (rad/mCi)
(rad) Adrenals 0.059 0.59 Brain 0.025 0.25 Breasts 0.026 0.26
Gallbladder wall 0.061 0.61 LLI wall 0.107 1.07 Small intestine
0.201 2.01 Stomach wall 0.048 0.48 ULI wall 0.221 2.21 Heart wall
0.109 1.09 Kidneys 0.078 0.78 Liver 0.096 0.96 Lungs 0.035 0.35
Muscle 0.037 0.37 Ovaries 0.088 0.88 Pancreas 0.063 0.63 Red marrow
0.037 0.37 Osteogenic cells 0.049 0.49 Skin 0.026 0.26 Spleen 0.079
0.79 Testes 0.041 0.41 Thymus 0.033 0.33 Thyroid 0.029 0.29 Urinary
bladder wall 0.666 6.66 Uterus 0.103 1.03 Total body 0.041 0.41
(rem/mCi) (rem) Effective dose 0.091 0.91
Example 2
Studies of .sup.18F-Labeled Aryl-Y-Alkylguanidine Compounds
[0164] During the development of the technology provided herein,
data were collected to characterize .sup.18F-labeled
aryl-Y-alkylguanidine compounds (in which Y is O, S, or NH) for use
as PET tracers. Experiments were conducted to evaluate closely
related analogs such as .sup.11C-labeled analogs of
phenoxyethylguanidine as imaging agents for cardiac sympathetic
innervation and adrenergic tumors in an isolated rat heart system
(Table 5).
TABLE-US-00005 TABLE 5 .sup.11C-labeled phenoxyethylguanidines Name
Structure (carbon-11 = *) .sup.11C-phenoxyethylguanidine
(.sup.11C-PEG) ##STR00057##
.sup.11C-meta-hydroxy-phenoxyethylguanidine (.sup.11C- MHPEG)
##STR00058## .sup.11C-4-fluoro-phenoxyethylguanidine
(.sup.11C-4F-PEG) ##STR00059##
[0165] C-11 compounds are generally easier to synthesize than F-18
compounds. However, F-18 compounds demonstrate better imaging
characteristics. As such, for some studies, C-11 compounds are
first synthesized and tested, and then promising candidates are
synthesized using F-18 rather than C-11. Experiments testing C-11
compounds at least demonstrate that a particular class of compounds
has members that function well as imaging agents. Then, effective
C-11 compounds having a ring fluorine indicate that placing an F-18
into that particular location will also produce an effective
compound with improved imaging characteristics. As such, the use of
C-11 labeled analogs is an efficient tool for screening compounds
that could potentially be F-18 labeled.
[0166] Experiments were conducted to test
.sup.11C-phenoxyethylguanidine (PEG), e.g., to establish baseline
values for the NET transport rate (K.sub.up, expressed in units of
mL perfusate/min/g wet) and the clearance rate from sympathetic
neurons (T.sub.1/2, expressed in units of hours). See FIG. 12. This
compound had a neuronal uptake rate of 5.37 mL/min/g wet, which is
amongst the fastest observed in any experiment, and cleared with a
major half-time of 5.7 hours.
[0167] Next, experiments tested a ring-fluorine analog,
.sup.11C-4-fluoro-phenoxyethylguanidine (4F-PEG), to assess the
influence of a fluorine on the neuronal uptake and clearance
kinetics in the isolated rat heart. 4F-PEG had a faster transport
rate that the PEG analog, viz., 7.25 mL/min/g wet, and cleared with
a major half-time of 3.5 hours (FIG. 12). The
4-[.sup.18F]fluoro-phenoxyethylguanidine (4-[.sup.18F]F-PEG) is
anticipated and contemplated to have nearly identical uptake and
clearance, thus providing attractive characteristics for an imaging
agent. It is further contemplated that the methods provided herein
are applicable to synthesize
4-[.sup.18F]fluoro-phenoxyethylguanidine.
[0168] A .sup.11C-meta-hydroxy-phenoxyethylguanidine (MHPEG) was
also tested to evaluate if a small structural change in the
compound led to efficient vesicular trapping, as had been observed
previously in studies with a series of phenethylguanidine
compounds. As predicted, this structural change led to complete
trapping of MHPEG (FIG. 12), while still retaining a rapid neuronal
uptake rate of 5.37 mL/min/g wet.
[0169] Based on the results with
.sup.11C-4-fluoro-meta-hydroxyphenethylguanidine
(.sup.11IC-4F-MHPG) and
4-[.sup.18F]fluoro-meta-hydroxy-phenethylguanidine
([.sup.18F]4F-MHPG), it was predicted that the corresponding
analogs, .sup.11C-4-fluoro-meta-phenoxyethylguanidine
(.sup.11C-4F-MHPEG) and
4-[.sup.18F]fluoro-meta-hydroxy-phenoxyethylguanidine
(4-[.sup.18F]-MHPEG) would likely be tracers with rapid NET
transport and efficient storage in vesicles. To test this
prediction, .sup.11C-4F-MHPEG and its positional isomer,
.sup.11C-3-fluoro-para-hydroxy-phenoxyethylguanidine
(11C-3F-PHPEG), were synthesized. Isolated rat heart studies with
these two agents demonstrated rapid uptake rates with very long
retention times (FIG. 13). .sup.11C-4F-MHPEG had a neuronal uptake
rate of 2.66 mL/min/g wet and was effectively trapped inside
storage vesicles. .sup.11C-3F-PHPEG had a faster uptake rate of
4.26 mL/min/g wet and cleared with a major half time of 16.9
hour.
[0170] Because of their rapid uptake rates and long retention
times, these compounds (e.g., labeled with either carbon-11 or
fluorine-18) are contemplated to find use as PET imaging agents,
e.g., for localizing adrenergic tumors. Additional biological
studies may reveal other uses of these imaging agents. According to
the technology provided herein, it is straightforward to use the
new .sup.18F-labeling methods described herein to prepare
4-[.sup.18F]-MHPEG, as well as other compounds in this class.
Example 3
Studies of Guanoxan Compounds
[0171] During the development of embodiments of the technology
provided herein, .sup.11C-guanoxan (.sup.11C-GOX) was synthesized
and experiments were conducted to evaluate its kinetics in the
isolated rat heart (Table 6). In addition, data were acquired in
studies of the kinetics of two ring-hydroxylated analogs of
guanoxan: .sup.11C-7-hydroxy-guanoxan (.sup.11C-7H-GOX) and
.sup.11C-6-hydroxy-guanoxan (.sup.11C-6H-GOX) (FIG. 14). Of the two
ring hydroxylated analogs, the 7-hydroxy analog demonstrated a more
rapid uptake and a longer retention time.
TABLE-US-00006 TABLE 6 .sup.11C-labeled guanoxans Name Structure
(carbon-11 = *) .sup.11C-guanoxan (.sup.11C-GOX) ##STR00060##
.sup.11C-7-hydroxy-guanoxan (.sup.11C-7H-GOX) ##STR00061##
.sup.11C-6-hydroxy-guanoxan (.sup.11C-7H-GOX) ##STR00062##
[0172] All publications and patents mentioned in the above
specification are herein incorporated by reference in their
entirety for all purposes. Various modifications and variations of
the described compositions, methods, and uses of the technology
will be apparent to those skilled in the art without departing from
the scope and spirit of the technology as described. Although the
technology has been described in connection with specific exemplary
embodiments, it should be understood that the technology as claimed
should not be unduly limited to such specific embodiments. Indeed,
various modifications of the described modes for carrying out the
technology that are obvious to those skilled in pharmacology,
biochemistry, medical science, or related fields are intended to be
within the scope of the following claims.
* * * * *