U.S. patent application number 11/517676 was filed with the patent office on 2007-01-11 for boat tropanes.
This patent application is currently assigned to President and Fellows of Harvard College. Invention is credited to Paul Blundell, Alan J. Fischman, Alun G. Jones, Bertha K. Madras, Ashfaq Mahmood, Peter C. Meltzer.
Application Number | 20070009432 11/517676 |
Document ID | / |
Family ID | 22460191 |
Filed Date | 2007-01-11 |
United States Patent
Application |
20070009432 |
Kind Code |
A1 |
Meltzer; Peter C. ; et
al. |
January 11, 2007 |
Boat tropanes
Abstract
Radiopharmaceutical compounds are disclosed. A tropane compound
is linked through the N atom at the 8-position to a chelating
ligand capable of complexing technetium or rhenium to produce a
neutral labeled complex that selectively binds to the dopamine
transporter over the serotonin transporter with a ratio of 10 or
more. These compounds can be prepared as separate diastereoisomers
as well as a mixture of diastereoisomers. Also disclosed are
radiopharmaceutical kits for preparing the labeled
radiopharmaceutical compounds.
Inventors: |
Meltzer; Peter C.;
(Lexington, MA) ; Blundell; Paul; (Winchester,
MA) ; Madras; Bertha K.; (Newton, MA) ;
Fischman; Alan J.; (Boston, MA) ; Jones; Alun G.;
(Newton Centre, MA) ; Mahmood; Ashfaq; (Newton
Center, MA) |
Correspondence
Address: |
EDWARDS & ANGELL, LLP
P.O. BOX 55874
BOSTON
MA
02205
US
|
Assignee: |
President and Fellows of Harvard
College
Cambridge
MA
02138
Organix, Inc.
Woburn
MA
01801
General Hospital Corporation, The
Boston
MA
02114
|
Family ID: |
22460191 |
Appl. No.: |
11/517676 |
Filed: |
September 8, 2006 |
Related U.S. Patent Documents
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Application
Number |
Filing Date |
Patent Number |
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09975586 |
Oct 11, 2001 |
7105678 |
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11517676 |
Sep 8, 2006 |
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09568106 |
May 10, 2000 |
6548041 |
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09975586 |
Oct 11, 2001 |
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09875523 |
Jun 6, 2001 |
6670375 |
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09975586 |
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09671534 |
Sep 27, 2000 |
6417221 |
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09875523 |
Jun 6, 2001 |
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09314441 |
May 19, 1999 |
6353105 |
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09671534 |
Sep 27, 2000 |
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08893921 |
Jul 11, 1997 |
5948933 |
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09314441 |
May 19, 1999 |
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08552584 |
Nov 3, 1995 |
6171576 |
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09671534 |
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60133761 |
May 12, 1999 |
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Current U.S.
Class: |
424/1.11 ;
534/14; 546/124 |
Current CPC
Class: |
A61K 51/0497 20130101;
A61P 25/28 20180101; A61B 6/506 20130101; A61P 25/00 20180101; A61P
43/00 20180101; A61K 51/0478 20130101; C07D 451/02 20130101; A61K
51/0448 20130101; A61K 31/46 20130101; C07F 13/005 20130101 |
Class at
Publication: |
424/001.11 ;
534/014; 546/124 |
International
Class: |
A61K 51/00 20060101
A61K051/00; C07F 13/00 20060101 C07F013/00; C07D 451/02 20060101
C07D451/02 |
Claims
1. A radiopharmaceutical compound which is capable of complexing
with .sup.99mTc, said compound having the following structural
formula: ##STR18## wherein R.sub.1 is .alpha. or .beta. and is
selected from COOR.sup.a, COR.sup.a, and CON(CH.sub.3)OR.sup.a;
R.sub.2 is .alpha. or .beta. and is selected from the group
consisting of C.sub.6H.sub.4X, C.sub.6H.sub.3XY, C.sub.10H.sub.7X,
and C.sub.10H.sub.6XY; R.sup.a is C.sub.1-C.sub.5 alkyl; X and Y
are independently selected from the group consisting of R.sup.a, H,
Br, Cl, I, F, OH, and OCH.sub.3; L is --(CH.sub.2).sub.n where n is
an integer from 1 to 6, or --(CH.sub.2).sub.n--(aryl, arylalkyl,
ethenyl or ethynyl)-(CH.sub.2).sub.m -where m and n are integers
and the sum of n plus m is an integer from 1 to 6; and Ch is a
tridentate or tetradentate chelating ligand that forms a neutral
complex with technetium or rhenium.
2. A compound according to claim 1 labeled with a radionuclide that
is complexed with the chelating ligand.
3. A compound according to claim 2, wherein the radionuclide is
.sup.99mTc.
4. A compound according to claim 2, wherein the radionuclide is
rhenium.
5. A compound according to claim 1, wherein the tropane analog has
a 3.alpha.-group.
6. A compound according to claim 1, wherein the tropane analog has
a 3.beta.-group.
7. A compound according to claim 1, wherein the chelating ligand
comprises a bisamido-bisthiol group, a monoamide,
monoamino-bisthiol group or a bisamino-bisthiol group covalently
attached to linker L.
8. (canceled)
9. A compound according to claim 1, wherein the chelating ligand is
N-(2-((2-((triphenylmethyl)thio)-ethyl)amino)acetyl)-S-(triphenylmethyl)--
2-aminoethanethiol.
10.-17. (canceled)
18. A method for detecting the density of tropane recognition sites
in a mammal as an indication of neurodegenerative or
neuropsychiatric disorders characterized by changes in the density
of dopamine transporters or dopamine neurons, said method
comprising providing in a suitable pharmacological carrier a
radiopharmaceutical compound according to claim 1 labeled with
.sup.99mTc, injecting the compound into the mammal and scanning the
mammal using a radiodiagnostic imaging apparatus.
19. A method for monitoring in a mammal neurodegenerative or
neuropsychiatric disorders characterized by changes in the density
of dopamine transporters or dopamine neurons, said method
comprising providing in a suitable pharmacological carrier a
radiopharmaceutical compound according to claim 1 labeled with
.sup.99mTc, injecting the compound into the mammal and scanning the
mammal using a radiodiagnostic imaging apparatus.
20. A radiopharmaceutical kit for preparing a radiopharmaceutical
preparation, said kit comprising a sealed, sterile, apyrogenic vial
containing a radiopharmaceutical compound of claim 1 and a reducing
agent for labeling said compound with a radionuclide.
21. (canceled)
22. A compound having the following structural formula: ##STR19##
wherein R.sub.1 is .alpha. or .beta. and is selected from
COOR.sup.a, COR.sup.a, and CON(CH.sub.3)OR.sup.a; R.sub.2 is
.alpha. and is selected from C.sub.6H.sub.4X, C.sub.6H.sub.3)(Y,
C.sub.10H.sub.7X, and C.sub.10H.sub.6XY; R.sup.a is a
C.sub.1-C.sub.5 alkyl; X and Y are independently selected from
R.sup.a, H, Br, Cl, I, F, OH, and OCH.sub.3; wherein the compound
is in the 1R or 1S configuration; and wherein N.sub.8 is
substituted with either H or CH.sub.3.
Description
FIELD OF THE INVENTION
[0001] The present invention relates to coordination complexes
comprising a radiolabeled ligand with high binding affinity and
good selectivity for the dopamine transporter (DAT). Such agents
can be useful for the early diagnosis and treatment of
neurodegenerative disorders.
BACKGROUND OF THE INVENTION
[0002] The dopamine transporter (DAT) plays a critical role in
physiological, pharmacological and pathological processes in brain.
The transport system is a primary mechanism for terminating the
effects of synaptic dopamine, thereby contributing to the
maintenance of homeostasis in dopamine systems. It also appears to
be a principal target of cocaine in the brain. (Kennedy and
Hanbauer, J. Neurochem. 1983, 41, 172-178; Shoemaker et al.,
Naunyn-Schmeideberg's Arch. Pharmacol. 1985, 329, 227-235; Reith et
al., Biochem Pharmacol. 1986, 35, 1123-1129; Ritz et al., Science
1987, 237, 1219-1223; Madras et al., J. Pharmacol. Exp. Ther.
1989a, 251, 131-141; Bergman et al., J. Pharmacol. Exp. Ther. 1989,
251, 150-155; Madras and Kaufman, Synapse 1994, 18, 261-275).
Furthermore, the dopamine transporter may be a conduit for entry of
neurotoxins into dopamine containing cells.
[0003] The striatum has the highest levels of dopamine terminals in
the brain. A high density of DAT is localized on dopamine neurons
in the striatum and appears to be a marker for a number of
physiological and pathological states. For example, in Parkinson's
disease, dopamine is severely reduced and the depletion of DAT in
the striatum has been an indicator for Parkinson's disease
(Schoemaker et al., Naunyn-Schmeideberg's Arch. Pharmacol. 1985,
329, 227-235; Kaufman and Madras, Synapse 1991, 9, 43-49).
Consequently, early or presymptomatic diagnosis of Parkinson's
disease can be achieved by the quantitative measurement of DAT
depletion in the striatum. (Kaufman and Madras, Synapse 1991, 9,
43-49). Simple and noninvasive methods of monitoring the DAT are
quite important. Depletion could be measured by a noninvasive means
such as brain imaging using a scintillation camera system and a
suitable imaging agent (Frost et al., Ann. Neurology 1993, 34,
423-431; Hantraye et al., Neuroreport 1992, 3, 265-268). Imaging of
the dopamine transporter also would enable the monitoring of
progression of the disease and of reversal of the disease such as
with therapies consisting of implants of dopamine neurons or drugs
that retard progression of the disease.
[0004] Other neuropsychiatric disorders, including Tourette's
Syndrome and Lesch Nyhan Syndrome and possibly Rett's syndrome, are
also marked by changes in DAT density. The DAT also is the target
of the most widely used drug for attention deficit disorder,
methylphenidate. The capacity to monitor the transporter in persons
suffering from this disorder can have diagnostic and therapeutic
implications. Furthermore, an age-related decline in dopamine
neurons can be reflected by a decline in the dopamine transporter
(Kaufman and Madras, Brain Res. 1993, 611, 322-328; van Dyck et
al., J. Nucl. Med. 1995, 36, 1175-1181) and may provide a view on
dopamine deficits that lie outside the realm of neuropsychiatric
diseases.
[0005] The density of the DAT in the brains of substance abusers
has also been shown to deviate from that in normal brain. For
example, the density is elevated in post-mortem tissues of cocaine
abusers (Little et al., Brain Res. 1993, 628, 17-25). On the other
hand, the density of the DAT in chronic nonviolent alcohol abusers
is decreased markedly. (Tiihonen et al., Nature Medicine 1995, 1,
654-657). Brain imaging of substance abusers can be useful for
understanding the pathological processes of cocaine and alcohol
abuse and monitoring restoration of normal brain function during
treatment.
[0006] Accordingly, a radiopharmaceutical that binds to the DAT can
provide important clinical information to assist in the diagnosis
and treatment of these various disease states.
[0007] In order to be effective as an imaging agent for the
disorders described above, it must have a specific binding affinity
and selectivity for the transporter being targeted, e.g. DAT. Brain
imaging agents must also have blood brain barrier (BBB)
permeability. Yet, it has been difficult to produce a metal chelate
which can cross the blood brain barrier while still retaining
binding affinity and selectivity for its receptor site. Therefore,
it is very desirable to find a suitable agent that satisfies these
criteria and will complex with a desired radionuclide, such as
.sup.99mTc.
[0008] In addition, to be an effective imaging agent, a specific
target:nontarget ratio is necessary. In the case of an agent
selective for DAT one must take into account the fact that the
striatum, the region of the brain having the highest density of the
dopamine transporter, also contains serotonin transporter (SET).
The SET is normally present at one-tenth to one-fifteenth the
concentration of the dopamine transporter. Imaging agents that bind
very strongly to DAT sometimes also exhibit a degree of binding to
SET. Although such a nontarget binding typically poses no serious
problem in the imaging of normal brains due to the greater number
of DAT compared to SET, under disease conditions in which DAT are
selectively reduced (or in which SET may be selectively increased),
binding to the SET may make it difficult to quantify DAT. Moreover,
binding to SET in other brain regions such as the hypothalamus and
thalamus can reduce striatal contrast and diminish accuracy in
localizing and imaging the striatum. Therefore, the target to
nontarget binding ratio of DAT:SET can be important. Presently,
among the most effective compounds for viewing and quantifying the
DAT are phenyltropane derivatives that are labelled with positron
emitters, such as .sup.11C and .sup.18F, and gamma emitters, such
as 123I.
[0009] The radionuclide, technetium-99m, .sup.99mTc (T.sub.1/26.9
h, 140 KeV gamma ray photon emission) is a preferred radionuclide
for use in imaging because of its excellent physical decay
properties and its chemistry. For example, its half-life of about 6
hours provides an excellent compromise between rate of decay and
convenient time frame for an imaging study. Thus, it is much
preferred to other radionuclides such as .sup.123I, which has a
substantially longer half life, or .sup.18F, which has a
substantially shorter half-life, and which are much more difficult
to use. Its emission characteristics also make it easy to image.
Further, it can be conveniently generated at the site of use.
.sup.99mTc is currently the radionuclide of choice in diagnostic
centers around the world. It would be desirable to have a
coordination complex with technetium for imaging DAT. Such a
complex could be used for detecting conditions in which the DAT is
useful as a marker.
[0010] However, a number of difficulties arise in the use of
technetium for radioimaging agents because of its chemistry. For
example, .sup.99mTc must typically be bound by a chelating agent.
Consequently it is much more difficult to design and prepare a
.sup.99mTc radioligand than it is to prepare a radioligand using
other radionuclides such as .sup.123I, which can be attached
covalently to the ligand. The size of the chelating agent for
technetium also can create problems when using this radionuclide in
imaging agents. This can be an especially difficult problem when
attempting to design receptor-based imaging agents using Tc.
[0011] Imaging agents being tested to determine their ability as
diagnostic tools for neurodegenerative diseases typically are
.sup.123I labeled radioiodinated molecules. See, for example,
RTI-55 (Boja, J. W., et al., Eur. J. Pharmacol. 1991, 194, 133-134;
Kaufman and Madras, Synapse, 1992, 12, 99-111) or .beta.-CIT
(Neumeyer, J. L., et al., Med. Chem. 1991, 34, 3144-3146) and an
iodoallyltropane, altropane (Elmaleh, D. R., et al., U.S. Pat. No.
5,493,026.
[0012] Although the tropane family of compounds are known to bind
to the dopamine transporter, the addition of bulky chelating
ligands for binding technetium or rhenium would be expected to
affect potency and ability to cross the blood brain barrier of the
resulting labeled complex. Kung, et al., in Technetium and Rhenium
in Chemistry and Nuclear Medicine 4, eds. M. Nicolini, G. Bandoli,
U. Mazzi, Servizi Grafici Editoriali, Padua, 1995, report that a
.sup.99Tc-labelled N.sub.2S.sub.2 ligand complexed with an
arylpiperazine known to have selective binding to serotonin.sub.1A
had only moderate binding affinity in vitro and failed to penetrate
the intact blood-brain barrier.
[0013] It would be desirable to have a technetium or rhenium
radio-labelled DAT imaging agent which is capable of crossing the
blood brain barrier and has a high binding affinity and selectivity
for the DAT.
SUMMARY OF THE INVENTION
[0014] The present invention provides radiopharmaceutical compounds
that form coordination complexes with a technetium or rhenium
radionuclide and that selectively bind to dopamine transporters,
thereby providing novel radio labeled agents. Preferred such agents
include radioimaging agents which are capable of crossing the blood
brain barrier to image DAT in the brain.
[0015] The compounds of the present invention comprise a tropane
compound linked through the N atom at the 8-position to a chelating
ligand capable of complexing a technetium or rhenium radionuclide
to produce a neutral labeled complex that selectively binds to the
dopamine transporter. These compounds can be prepared as separate
diastereoisomers as well as a mixture of diastereoisomers.
[0016] Tropane compounds useful in the practice of the present
invention bind to the dopamine transporter. Preferred
radiopharmaceutical compounds of the invention can be represented
by the following structural formula: ##STR1## wherein [0017]
R.sub.1 is .alpha. or .beta. and is selected from COOR.sup.a,
COR.sup.a, and CON(CH.sub.3)OR.sup.a; [0018] R.sub.2 is .alpha. or
.beta. and is selected from C.sub.6H.sub.4X, C.sub.6H.sub.3XY,
C.sub.10H.sub.7X, and C.sub.10H.sub.6XY; [0019] R.sup.a is selected
from C.sub.1-C.sub.5 alkyl, e.g. methyl, ethyl, propyl, isopropyl,
etc.; [0020] X and Y are independently selected from R.sup.a, H,
Br, Cl, I, F, OH, and OCH.sub.3;
[0021] L is --(CH.sub.2).sub.n where n is an integer from 1 to 6,
or --(CH.sub.2).sub.n-(aryl, arylalkyl, ethenyl or
ethynyl)-(CH.sub.2).sub.m where the sum of n plus m is an integer
from 1 to 6; and [0022] Ch is a tridentate or tetradentate
chelating ligand that forms a neutral complex with technetium or
rhenium, and further wherein the bond between C.sub.2 and C.sub.3
is either a single bond or a double bond. Thus, R.sub.1 and R.sub.2
can be in the .alpha. or .beta. configuration. Further, R.sub.1
preferably can be substituted at the C.sub.2 or C.sub.4 when the
tropane has a 1R or 1S configuration, respectively. The chelating
ligand, if chiral, can be syn or anti with R, S, or RS.
[0023] The imaging agents of the present invention are useful for
detecting tropane recognition sites including neuronal transporters
such as the dopamine transporter. For purposes of the present
invention, a tropane recognition site is any receptor or
transporter site that binds to the tropane compound. Thus, the
compounds of this invention can be used as diagnostic agents,
prognostic agents and therapeutic agents for neurodegenerative
diseases.
[0024] The present invention also provides a method of using the
coordination complex as an imaging agent for detecting
neurodegenerative and neuropsychiatric disorders characterized by a
change in density of DAT or dopamine neurons. For example, a method
for detecting the change in DAT resulting from a neurodegenerative
disease, such as Parkinson's disease, comprises injecting a labeled
compound of the present invention in a dose effective amount for
detecting DAT in the particular mammal and obtaining images of the
labeled compound bound to DAT. Rhenium labeled compounds can also
be useful for therapeutic treatments.
[0025] The present invention also provides kits for producing the
compounds of the present invention labeled with technetium or
rhenium. The kits typically comprise a sterile, non-pyrogenic
container containing lyophilized compound and a reducing agent to
form a complex of the compound with technetium or rhenium. The kits
permit ready reconstitution and labeling with aqueous solutions
containing the radionuclide, e.g. pertechnetate, preferably having
a pH in the range of about 5 to about 8.
BRIEF DESCRIPTION OF THE DRAWINGS
[0026] FIG. 1 is an illustration of alternative general reaction
schemes for linking a N.sub.2S.sub.2 chelating ligand to a
nortropane analog and labeling with a metal "M", which can be
technetium or rhenium.
[0027] FIG. 2 is an illustration of a general scheme for
preparation of 2-carbomethoxy tropanes (Scheme 1) comprising an
aryl octene in accord with the present invention and subsequent
preparation of 3.alpha. and 3.beta. diasteriomers thereof.
[0028] FIGS. 3 and 4 are illustrations of a general scheme for
preparation of 2-ethylketo analogs (Schemes 2 and 3) of
radiopharmaceutical compounds in accord with a preferred embodiment
of the present invention.
[0029] FIG. 5 is an illustration of a general scheme for
preparation of 2-carboxamido 3.alpha.- or 3.beta.-aryl analogs
(Scheme 4) of radiopharmaceutical compounds in accord with a
preferred embodiment of the present invention.
[0030] FIG. 6 is an illustration of a general reaction scheme for
preparation of 3-aryl-2-ethylketo-2,3-ene analogs (Scheme 5) of
radiopharmaceutical compounds in accord with a preferred embodiment
of the present invention.
[0031] FIG. 7 is an illustration of a general reaction scheme for
preparation of 3-aryl-2-carbomethoxy-2,3-ene analogs (Scheme 6) of
radiopharmaceutical compounds in accord with a preferred embodiment
of the present invention.
[0032] FIG. 8 is an illustration of a general scheme for converting
the 3.alpha. and 3.beta. diasteriomers of FIG. 2 to bistrityl
protected N.sub.2S.sub.2 tropanes and labeling with technetium or
rhenium (Scheme 7).
[0033] FIG. 9 is an illustration of an alternative general reaction
scheme for preparation of 3-aryl-2-ethylketo-2,3-ene analogs
(Scheme 8) of radiopharmaceutical compounds in accord with a
preferred embodiment of the present invention.
[0034] FIG. 10 is an illustration of a general reaction scheme for
preparation of 3-naphthyl-2-carbomethoxy-2,3-ene and
3.alpha.-naphthyl or 3.beta.-naphthyl analogs (Scheme 9) of
radiopharmaceutical compounds in accord with a preferred embodiment
of the present invention.
[0035] FIG. 11 is a HPLC chromatogram of .sup.99mTc labeled
0-1505T.
[0036] FIG. 12 is a HPLC chromatogram of .sup.99mTc labeled
0-1508T.
[0037] FIG. 13 is a HPLC chromatogram of .sup.99mTc labeled
0-1561T.
[0038] FIG. 14 is a HPLC chromatogram of .sup.99mTc labeled
0-1560T.
DETAILED DESCRIPTION OF THE INVENTION
[0039] The compounds of the present invention comprise a tropane
compound or ligand that selectively binds to tropane recognition
sites, e.g., neuron transporters such as the DAT. The tropane
ligand is radiolabeled with a radioactive technetium or rhenium by
a chelating ligand which is attached to the tropane ligand by a
linker. The unlabeled compounds of this invention are schematically
represented by the formula Ch--L--Tr, wherein Ch is the chelating
ligand, L is the linker and Tr is the tropane ligand.
[0040] Tropane compounds or ligands useful in the practice of the
present invention can generally be represented by formula II where
R.sub.1 and R.sub.2 are defined as above and where R.sub.1 can also
be substituted at the C.sub.4 position of the tropane ring:
##STR2##
[0041] Any tropane compound of the general formula II is useful in
the present invention so long as it binds to DAT. Examples of
particularly useful tropanes are:
2-carbomethoxy-3-(4-fluorophenyl)-N-methyltropane ("WIN 35,428")
(Clarke, R. L., et al., J. Med. Chem. 1973, 16, 1260-1267) which
binds potently (IC.sub.50=11.0 nM) and with specificity to the DAT
(Meltzer, P. C., et al., J. Med. Chem. 1993, 36, 855-862);
2-carbomethoxy-3-(3,4-dichlorophenyl)-N-methyltropane ("O-401";
IC.sub.50=1.09 nM) (Meltzer, P. C., et al., J. Med. Chem. 1993, 36,
855-862). Tropane analogs that have a 3.alpha.-group are of the
boat configuration. Other tropanes having a 3.beta.-oriented group
are of the chair configuration.
[0042] Chelating ligands useful in the practice of the present
invention comprise any tridentate or tetradentate ligand that binds
technetium or rhenium to form a neutral complex. The chelating
ligand is covalently attached to the linker L, as described below.
Preferred chelating ligands contain a plurality of N or S atoms for
complexing with the radionuclide.
[0043] Examples of suitable ligands are the N.sub.2S.sub.2
compounds represented by the following structural formulas:
##STR3## wherein R, R.sup.6, and R.sup.10 are each selected from
hydrogen, substituted or unsubstituted lower alkyl, alkylR.sup.9,
or --COR.sup.9 where R.sup.9 is selected from hydroxy, substituted
lower alkoxy, substituted or unsubstituted amino, glycine ester,
halide (chloro, bromo, iodo) or OR (OR is a leaving group such as
mesylate, triflate, or tosylate) or an activated leaving group;
R.sup.1 is selected from hydrogen, or substituted or unsubstituted
lower alkyl; R.sup.2 and R.sup.3 are each selected from hydrogen or
a thiol protecting group, or an inter or intramolecular disulfide;
and R.sup.4, R.sup.5, R.sup.7 and R.sup.8 are each selected from
hydrogen or lower alkyl.
[0044] When R, R.sup.6 or R.sup.10 is a carboxylic acid derivative,
R.sup.9 can be an activated leaving group. For purposes of this
invention the leaving group R.sup.9 is defined such that
(compound)-COR.sup.9 is an acylating agent. Examples of activated
leaving groups suitable for the practice of this invention include,
for example: halide; substituted or unsubstituted aryloxy groups
such as phenoxy, pentachlorophenoxy, etc,; oxy-heterocyclic groups
such as N-oxy-succinimido, etc.; mercapto; lower alkylthio;
arylthio; oxyphosphonium; and other groups known to those skilled
in the art to be useful as leaving groups.
[0045] R.sup.2 and R.sup.3 can be hydrogen or any known thiol
protecting group. Examples of such groups include lower
alkylaminocarbonyl such as ethylaminocarbonyl, lower
alkanoylaminomethyl, aroylaminomethyl, t-butyl, acetamidomethyl,
arylmethyl such as triphenylmethyl (trityl) and diphenylmethyl,
aroyl such as benzoyl, aryloxycarbonyl such as phenoxycarbonyl,
arylloweralkoxycarbonyl, preferably arylmethoxycarbonyl,
benzyloxycarbonyl, and lower alkoxycarbonyl such as
t-butoxycarbonyl. Preferred thiol protecting groups include trityl,
t-butyl, diphenylmethyl, acetamidomethyl and benzoyl and an inter
or intramolecular disulfide.
[0046] The term "lower alkyl" when used herein designates aliphatic
saturated branched or straight chain hydrocarbon monovalent
substituents containing from 1 to 6 carbon atoms such as methyl,
ethyl, isopropyl, n-propyl, n-butyl, etc., more preferably 1 to 4
carbons. The term "lower alkoxy" designates lower alkoxy
substituents containing from 1 to 6 carbon atoms such as methoxy,
ethoxy, isopropoxy, etc., more preferably 1 to 4 carbon atoms.
[0047] The terms substituted lower alkyl or substituted lower
alkoxy when used herein include alkyl and alkoxy groups substituted
with halide, hydroxy, carboxylic acid, or carboxamide groups, etc.
such as, for example, --CH.sub.2OH, --CH.sub.2CH.sub.2COOH,
--CH.sub.2CONH.sub.2, --OCH.sub.2CH.sub.2OH, --OCH.sub.2COOH,
--OCH.sub.2CH.sub.2CONH.sub.2, etc.
[0048] The term substituted amino when used herein includes such
groups mono or di and tri-substituted with lower alkyl, and
--NH.sub.3.sup.+ or mono, di and tri-substituted ammonium groups
substituted with lower alkyl with a pharmacologically suitable
anion.
[0049] The term glycine ester as used herein means the lower alkyl
esters of glycine, preferably the methyl and ethyl esters.
[0050] These chelating ligands can be complexed with a
radionuclide, e.g., technetium, to form the following complexes:
##STR4## where the R groups are defined as above.
[0051] Preferred embodiments of the invention use chelating ligands
that are formed from monoaminomonoamide compounds having structures
of formula V, VI or VII, e.g.,
N-{2-((2-((triphenylmethyl)-thio)-ethyl)amino)acetyl}-S-(triphenylmethyl)-
-2-aminoethanethiol ("MAMA'").
[0052] Any organic linker having a backbone chain length of 1 to
about 6 carbon atoms can be used to attach the chelating ligand,
typically through its nitrogen, sulfur, R, R.sup.1 or R.sup.6, to
the 8-nitrogen atom of the tropane ligand (which binds the dopamine
transporter). Examples of linkers include --(CH.sub.2).sub.n where
n is an integer from 1 to 6, or --(CH.sub.2).sub.n-(aryl,
arylalkyl, ethenyl or ethynyl)-(CH.sub.2).sub.m where the sum of n
plus m is an integer from 1 to 6.
[0053] Preferred radiolabeled compounds of the present invention
cross the blood brain barrier and exhibit desired target:non-target
specificity. Preferably, the selectivity ratio of binding (DAT:SET)
is about 30 or more, more preferebly 50 or more. Thus, they are
useful as brain imaging agents, for example, for imaging DAT.
[0054] The tropane ligands can be linked to the chelating ligand by
an initial conversion to nortropanes. Syntheses of nortropanes are
known in the art, for example, as disclosed in Meltzer, P. C., et
al., J. Med. Chem. 1993, 36, 855-862; Meltzer, P. C., et al., J.
Med. Chem. 1994, 37, 2001-2010 (the disclosure of which is
incorporated herein by reference). Tropanes can be synthesized from
tropinone or cocaine by techniques known in the art. Synthesis of
the nortropanes can then be achieved by N-demethylation of the
tropane, which can be readily accomplished by various methods known
in the art, e.g., with .alpha.-chloroethyl chloro formate
(ACE-Cl).
[0055] The chelating ligand is preferably prepared separately and,
then, either attached to the nortropane and metallated, or
metallated first followed by attachment to the appropriate
nortropane. When the radiolabeled compounds of the invention are
required to cross the blood brain barrier, the chelating ligands
useful in the present invention form neutral complexes with the
radionuclide and are lipid soluble. Chelating ligands that form
neutral .sup.99mTc(V) complexes which are useful in the present
invention include a substituted oxime (Loberg, M. D., et al., J.
Nucl. Med. 1979, 20, 1181-1188), N.sub.2S.sub.2 compounds (Davison,
A., et al., Inorg. Chem. 1981, 20, 1629-1632; Davison, A., et al.,
J. Nucl. Med. 1979, 20, 641 (abstr)), bisaminoethanethiol ("BAT")
(Kung, H. F., et al., J. Med. Chem. 1985, 28, 1280-1284; Kung, H.
F., et al., J. Nucl. Med. 1986, 27, 1051; Kung, H. F., et al., J.
Med. Chem. 1989, 32, 433-437; Kung, H. F., et al., J. Nucl. Med.
1984, 25, 326-332; Francesconi, L. C., et al., Inorg. Chem. 1993,
32, 3114-3124), and diaminodithiol ("DADT") (Lever, S. Z., et al.,
J. Nucl. Med. 1985, 26, 1287-1294). Additional examples of useful
chelating ligands include
N,N'-bis(2-mercapto-1-methyl)-2-aminobenzylamine ("U-BAT")
(Francesconi, L. C., et al., J. Med. Chem. 1994, 37, 3282-3288),
propylene amine oximes ("HMPAO"), diamidodithiol ("DADS") (Rao, T.
N., et al., J. Am. Chem. Soc. 1990, 112, 5798-5804;
Stepniak-Biniakiewicz, D., et al., J. Med. Chem. 1992, 35,
274-279), phenylenediamine-thiol-thioether ("PhAT") (McBride, B.
J., et al., J. Med. Chem. 1993, 36, 81-86),
bis(mercaptoethyl)-2-aminoethylamine ("SNS") or
bis(mercaptoethyl)-2-thioethylamine (Mastrostamatis, S. G., et al.,
J. Med. Chem. 1994, 37, 3212-3218), monoamine amide ("MAMA")
(Gustavson, L. M., et al., Tet. Lett. 1991, 32, 5485-5488) and
N-{2-((2-((triphenylmethyl)thio)ethyl)amino)acetyl}-S-(triphenylmethyl)-2-
-aminoethanethiol ("MAMA"') (O'Neil, J. P., et al., Inorg. Chem.
1994, 33, 319-323). For example, when MAMA' is attached to a
lipophilic tropane by a linker in accord with the present
invention, a neutral, moderately lipophilic and aqueous stable
compound suitable for radiolabeling is formed.
[0056] Compounds of formula III and IV can be synthesized according
to the methods described in U.S. Pat. No. 4,673,562 which is
incorporated herein by reference. Compounds of formula V can be
synthesized by methods known in the art (see Fritzberg et al., J.
Nucl. Med. 1981, 22, 258-263). Compounds of formula VI can also be
synthesized by methods known in the art. (See O'Neil, J. P., et
al., Inorg. Chem. 1994, 33, 319-323).
[0057] Radiolabeled complexes of the present invention can be
prepared via three general preparation procedures as outlined in
the General Scheme (FIG. 1). The general preparation scheme
exemplifies the use of trityl protecting groups for the
sulfhydryls, however, other protecting groups that are known to be
useful for sulfhydryl protection can also be used such as, for
example, lower alkylaminocarbonyl such as ethylaminocarbonyl, lower
alkanoylaminomethyl, arylaminomethyl, t-butyl, acetamidomethyl,
arylmethyl such as triphenylmethyl(trityl) and diphenylmethyl, aryl
such as benzoyl, aryloxycarbonyl such as phenoxycarbonyl, aryl
loweralkoxycarbonyl, preferably arylmethoxycarbonyl such as
benzyloxycarbonyl, and lower alkoxycarbonyl such as
t-butoxycarbonyl. Preferred sulfhydryl protecting groups include
trityl, t-butyl, diphenylmethyl, acetamidomethyl, disulfide and
benzoyl.
[0058] The compounds of the invention can be prepared by known
means based upon the present disclosure. For example, starting with
an appropriate chelating ligand, such as an N.sub.2S.sub.2
compound, illustrated in the general scheme in FIG. 1 as
N-{2-((2-((triphenylmethyl)thio)ethyl)amino)acetyl}-S-(triphenylmethyl)-2-
-aminoethanethiol, (MAMA': Katzenellenbogen et al., Inorg. Chem.,
1994, 33, 319), the N.sub.2S.sub.2 compound can be alkylated with
either the haloalkyl triflate or the haloalkylnortropane (prepared
from the nortropane: Meltzer et al., J. Med. Chem., 1993, 36, 855),
producing the chloroalkyl (propyl shown) MAMA', or the tropanalkyl
(propyl shown) MAMA', compounds, respectively. The chloroalkyl
(propyl shown) MAMA' compound, can then be attached to a suitable
nortropane to provide the tropanalkyl (propyl shown) MAMA'
compounds, as shown. Alternatively, the chloroalkyl (propyl shown)
MAMA', can be treated to incorporate a metal atom, preferably a
radionuclide (such as .sup.99Tc, .sup.99mTc, .sup.188Re or
.sup.186Re) to provide the M-labeled complex. The resulting complex
can then be attached to a suitable nortropane to provide
radiopharmaceutical compounds of the present invention, as
shown.
[0059] Alternatively, the tropanalkyl (propyl shown) MAMA'
compounds, can be treated to incorporate a radionuclide (such as
.sup.99Tc, .sup.99mTc, .sup.188Re or .sup.186Re) to form
radiopharmaceutical compounds of the present invention, as
shown.
[0060] The compounds of the present invention can be either
diastereoisomer as well as a mixture of both diastereomers. The
diastereoisomers can be separated by column chromatography.
[0061] More specifically, alkylation of the N.sub.2S.sub.2, with
haloalkyl triflate to produce the chloroalkyl (propyl shown) MAMA',
can be used to prepare the linker which is used to bind the
chelating ligand to the tropane ligand, which selectively binds the
dopamine transporter. This alkylation step can be modified by those
of ordinary skill in organic chemistry to create various linkers
having a backbone chain length of 1 to about 6 carbon atoms, as
described above.
[0062] Deprotection of the chloroalkyl compound can be accomplished
by standard methods well known in the art, e.g., with
H.sub.2S/Hg(OAc).sub.2 (O'Neil, J. P., et al., Inorg. Chem. 1994,
33, 319-323) or AgNO.sub.3/Py (DiZio, J. P., et al., Bioconj. Chem.
1991, 2, 353-366), with TFA and phenol, or HBr in acetic acid
(Zervas, L., et al., J. Amer. Chem. Soc. 1962, 84, 3887-3897) to
result in the unprotected bisthiol which can then be immediately
treated with a solution of tin (II) chloride (SnCl.sub.2) and
sodium perrhenate (Na.sub.2ReO.sub.7) or an agent such as
Na(.sup.99mTcO.sub.4)/stannous tartrate (Francesconi, L. C., et
al., Inorg. Chem. 1993, 32, 3114-3124; Canney, D. J., et al., J.
Med. Chem. 1993, 36, 1032-1040) to produce the labeled complexes.
Purification of these chelates can be accomplished by flash
chromatography as described by O'Neil (O'Neil, J. P., et al.,
Inorg. Chem. 1994, 33, 319-323). The chloroalkyl chelate, can then
be reacted (O'Neil, J. P., et al., Bioconj. Chem. 1994, 5, 182-193)
with the appropriate nortropane to provide the labeled coordination
complexes of the present invention. Alkylation of nortropanes can
be accomplished by methods known in the art, e.g., acetonitrile
(CH.sub.3CN), potassium iodide (KI) and potassium carbonate
(K.sub.2CO.sub.3). The use of strong base can cause epimerization
of the carbomethoxy group at C-2, although sodium carbonate in a
solvent such as dimethyl formamide (DMF) can yield alkylated
products in reasonable yield.
[0063] These compounds can be prepared either as free bases or as a
pharmacologically active salt thereof such as hydrochloride,
tartrate, sulfate, naphthalene-1,5-disulfonate or the like.
[0064] Reaction schemes for preparation of various classes of
compounds of the present invention are described with reference to
the drawings. In Scheme 1, as illustrated in FIG. 2, Keto ester 1
{Meltzer et al., J. Med. Chem, 1994, 37, 2001} is converted to the
enol triflate 2 by reaction with
N-phenyltrifluoromethanesulfonimide and sodium
bis(trimethylsilyl)amide in tetrahydrofuran. The enol triflate 2 is
then coupled with the appropriate commercial or preformed
arylboronic acids by Suzuki coupling in diethoxymethane in the
presence of lithium chloride, sodium carbonate and
tris(dibenzylideneacetone)dipalladium(0) to provide aryl octenes 3
in excellent yield.
[0065] Reduction of the octenes 3 with samarium iodide in
tetrahydrofuran/methanol at low temperature (-78.degree. C.)
provides a mixture of the 3.beta.- and 3.alpha.-diastereomers, 4
and 12 respectively. These diastereomers are readily separated by
flash column chromatography.
[0066] In Scheme 7, as illustrated in FIG. 8, the 3.beta.- and
3.alpha.-diastereomers 4 and 12 are then treated similarly in their
conversion to the bistrityl protected N.sub.2S.sub.2 tropanes 33
and 36 respectively and thence to the rhenium analogs 34R and 37R
and the technetium analogs 34T and 37T. Thus, the tropanes are
N-dealkylated by treatment with ACE-chloride {Meltzer et al., J.
Med. Chem, 1993, 36, 855} to provide the nortropanes 32 and 35.
Introduction of the N.sub.2S.sub.2 protected, ligand is then
achieved by reaction of the nortropanes with the preformed
N-[[[2-[2-(triphenylmethyl)thio]ethyl](N'-3'-chloropropyl)amino]acetyl]-S-
-(triphenylmethyl)-2- aminoethanethiol (MAMA'-Cl){Meltzer et al. J.
Med. Chem., 40, 1835, 1997} in the presence of potassium iodide and
potassium carbonate. Rhenium can then be introduced upon reaction
with tin (II) chloride in 0.05 M HCl, followed by sodium perrhenate
in 0.05 M HCl. The product is purified by silica gel column
chromatography and obtained as a mixture of diastereomers. An
alternative approach utilizes N-alkylation of the nortropanes with
preformed N-[(2-((3'-chloropropyl)
(2-mercaptoethyl)amino)acetyl)-2-aminoethane-thiolato]rhenium (V)
oxide {Meltzer et al. J. Med. Chem., 40, 1835, 1997}. Both
diastereomers of the metal chelate are prepared.
[0067] In Schemes 2 and 3, as illustrated in FIGS. 3 and 4, the
2-ethylketo analogs in both the 3.alpha.- and 3.beta.-series are
prepared as shown. This discussion exemplifies the
3.beta.-diastereomer, as illustrated in FIG. 3. The
3.beta.-diastereomer 4 is hydrolyzed in dioxane/water or with
lithium hydroxide to provide the acid, 5 which is converted to the
amide through conversion to the acid chloride with oxalyl chloride
and then reaction with (MeO)MeNH.HCl to provide 6. Further reaction
with alkyl Grignard at low temperature provides the desired ethyl
or alkyl ketone 7. The alkyl ketone 7 can be made alternatively by
reaction of the ester-tropane (4 or 12) with the appropriate
alkyl-Grignard-(ethylmagnesium bromide in this case) (Ref: I.
Kikkawa and T. Vorifuji, Synthesis (1980), p. 877). Demethylation
is by standard treatment with ACE-Cl to obtain 8. The chelating
unit is attached by reaction of 8 with MAMA'-Cl in the presence of
a base such as potassium carbonate or potassium bicarbonate and
potassium iodide. Insertion of rhenium to provide 10 or technetium
to provide 11 is accomplished with sodium perrhenate under
reductive conditions, of the technetium heptogluconate. Both
diastereomers of the metal chelate can be prepared in a similar
manner. As seen by comparing Schemes 2 and 3, the
3.alpha.-diastereomer of the tropane is prepared in a similar
manner.
[0068] In Scheme 4, as illustrated in FIG. 5, the 2-carboxamide
analogs 22 and 22A are prepared from 14 by N-demethylation with
ACE-Cl as described earlier.
[0069] Attachment of the MAMA' group and insertion of the metal
(rhenium or technetium) is conducted as described earlier for
Schemes 2 and 3.
[0070] In Scheme 5, as illustrated in FIG. 6, the 2-ethylketo
trop-2-ene analogs are prepared as shown in the scheme. Thus, ester
3 is hydrolyzed in dioxane/water or with lithium hydroxide to
provide the acid 23 which is converted to the amide through
conversion to the acid chloride with oxalyl chloride and then
reaction with (MeO)MeNH.HCl to provide 24. Further reaction with
alkyl Grignard at low temperature provides the desired ethyl or
alkyl ketone 25. The alkyl ketone 25 can be formed altneratively by
reaction of the ester tropene (3) with the appropriate
alkyl-Grignard-(ethylmagnesium bromide) (Ref: I. Kikkawa and T.
Vorifuji, Synthesis (1980), p. 877). Demethylation is by standard
treatment with ACE-Cl to obtain 26. The chelating unit is attached
by reaction of 26 with MAMA'-Cl in the presence of a base such as
potassium carbonate or potassium bicarbonate and potassium iodide.
Insertion of rhenium or technetium to provide 28 is accomplished
with sodium perrhenate under reductive conditions of the technetium
heptogluconate. Both diastereomers of the metal chelate 28 are
prepared.
[0071] In Scheme 6, as illustrated in FIG. 7, the 2-carbomethoxy
trop-2-ene analogs are prepared as shown in the scheme.
Demethylation of compound 3 is by standard treatment with ACE-Cl to
obtain compound 29. The chelating unit is attached by reaction of
compound 30 with MAMA'-Cl in the presence of a base such as
potassium carbonate or potassium bicarbonate and potassium iodide.
Insertion of rhenium or technetium to provide compound 31 is
accomplished with sodium perrhenate under reductive conditions, of
the technetium heptogluconate. Both diastereomers of the metal
chelate 31 are prepared.
[0072] In Scheme 8, as illustrated in FIG. 9, the 2-ethylketo
trop-2-ene analogs are prepared as shown in the scheme. Thus,
compound 3 is reduced with LAH and reoxidized to obtain the
aldehyde 39. Reaction of the aldehyde with ethyl lithium or ethyl
Grignard provides the alcohol 40 which is oxidized once again to
obtain the ethyl ketone 26. Compounds 27 and 28 are then obtained
as above.
[0073] In Scheme 9, as illustrated in FIG. 10, the 3-naphthyl
trop-2-ene and 3.alpha.-and 3.beta.-tropanes are obtained by
similar chemistry to that described earlier. Thus, the trop-2-ene
38 is N-demethylated with ACE-Cl and the MAMA' is attached to
provide compound 40. Rhenium or technetium are inserted as before
to obtain the diastereomers 41. Alternatively, compound 38 is first
reduced with samarium iodide to obtain both the boat and chair
configured compounds 43 and 42. The same sequence of reactions then
provides the rhenium and technetium diastereomers of both the
3.alpha.- and 3.beta.-tropanes, 49 and 48.
[0074] The technetium or rhenium radionuclide complexes of this
invention can be formed by reacting suitable precursor compounds
with either pertechnetate or perrhenate in the presence of a
suitable reducing agent in a conventional manner. For example, the
compound can be dissolved in a suitable solvent with a reducing
agent and then pertechnetate added. The mixture is then heated for
a suitable length of time to complete the reaction. Typically,
heating in a boiling water bath for about 10 minutes has been found
sufficient to obtain very good yields of the radionuclide complex.
To form rhenium complexes, (Ph.sub.3P).sub.2ReOCl.sub.3 is added in
the presence of basic (NaOAc) methanol. Examples of reducing agents
useful in the practice of this invention include stannous salts
such as stannous chloride, sodium dithionite, and ferrous salts
such as ferrous sulfate.
[0075] Rhenium behaves similarly to Tc. Thus, N.sub.2S.sub.2
complexes of Re or Tc are equally stable. Both metals form square
pyramidal complexes with N.sub.2S.sub.2 ligands. (Francesconi, L.
C., et al., Inorg. Chem. 1993, 32, 3114-3124). Rhenium is a
preferred metal for use in studies which do not require the
presence of a short half life radiolabel. For complexes with both
technetium and rhenium, the oxygen occupies an apical position,
therefore both syn and anti-isomers of the metal complexes are
possible. The biological activity of Tc and Re chelates are
generally similar. (O'Neil, J. P., et al., Bioconjugate Chem. 1994,
5, 182-193). .sup.99mTc is a preferred radionuclide for use as an
imaging agent. Rhenium is an excellent model for .sup.99mTc and is
also useful as a therapeutic agent.
[0076] A preferred method for introducing the technetium
radionuclide was by reaction of the bistrtityl protected compounds
in presence of anhydrous trifluoroacetic acid and then
triethylsilane. A portion of the aqueous solution thus obtained was
then incubated with .sup.99mTc-glucoheptonate solution (Glucoscan
kits from Du Pont, Billerica, Mass.). HPLC separation on a C.sub.8
reverse phase column equipped provides the major .sup.99mTc labeled
product which was reconstituted in sterile saline for
injection.
[0077] The compounds of this invention are typically
enantiomerically pure tropanes (either 1S or 1R configuration)
attached by an achiral linker to a chiral chelating ligand. The
chiral chelating ligand can be cis or trans with respect to the
metal oxo and the linker, but is preferably cis. Each of the cis
and trans chelating ligands exist as a pair of two enantiomers. By
virtue of the chiral ligand which can exist in each of two
enantiomeric forms, and a chiral tropane, each of the whole
molecules exists as diastereoisomers. Radiopharmaceutical
compositions of the present invention include the separate
diastereoisomers of each, as well as mixtures of diastereomeric
pairs.
[0078] The compounds of the present invention preferably have a
target:nontarget ratio, such as a DAT:SET selectivity ratio of
greater than 10, and preferably at least 30, to minimize binding of
trace levels of the drug to the nontarget, e.g., serotonin
transporter.
[0079] The present invention also provides pharmaceutical kits,
preferably comprising the compounds of formula I with a reducing
agent in lyophilized form in a pyrogen-free, sterilized container
or vial. In this form the lyophilized composition can be readily
reconstituted by adding only water, saline, or a buffer preferably
having a pH in the range of 5 to 8, more preferably physiological
pH. If technetium is the metal to be used as the radionuclide,
pertechnetate solution from a technetium generator can be used for
reconstitution.
[0080] In general, the radiopharmaceutical preparation kit
comprises a sterilized unit dose (or multidose) vial containing the
purified compound of formula I and a reducing agent for technetium,
preferably lyophilized. Each dose should consist of a sufficient
amount of compound and reducing agent to prepare the required dose
for imaging, normally about 5 to about 30 mCi of .sup.99mTc
depending upon body weight of the mammal to be imaged. In use, the
technetium, preferably as .sup.99mTc-pertechnetate in saline, is
injected aseptically into the vial and the mixture reacted for a
sufficient time to form the labeled complex. After reaction,
typically, the resulting radiopharmaceutical is ready for use.
[0081] To image a desired target, a radiopharmaceutical preparation
in accord with this invention having an effective dose of
radioactivity for the particular mammal is prepared in a suitable
pharmacological carrier, such as normal saline. Preferably, the
radiopharmaceutical preparation is injected intravenously into the
mammal. The target, e.g., the brain, is then imaged by positioning
the mammal under a gamma camera or other suitable device.
[0082] In order to obtain high quality images, the radiochemical
yield of bound technetium in the desired radiopharmaceutical should
preferably be greater than 70% after reconstituting the lyophilized
mixture and labelling. Lower yields may result in poorer image
quality and undesirable purification steps may be required to
produce high quality images.
[0083] This invention will be illustrated further by the following
examples. These examples are not intended to limit the scope of the
claimed invention in any manner.
[0084] The final compounds were characterized and their purity
analyzed prior to biological evaluation. High field nuclear
magnetic resonance (NMR) spectra were measured as well as low and
high resolution mass spectra (MS) and infrared spectra (IR).
Elemental analyses, thin layer chromatography (TLC) and/or high
performance liquid chromatography (HPLC) were used as a measure of
purity. A purity of >98% was obtained before biological
evaluation of these compounds was undertaken.
[0085] In the following examples, NMR spectra were recorded on
either a Bruker 100, a Varian XL 400, or a Bruker 300, or a Jeol
300 NMR spectrometer. TMS was used as internal standard. Melting
points are uncorrected and were measured on a Gallenkamp melting
point apparatus. Optical rotations were measured at the sodium D
line at 21.degree. C. using a JASCO DIP 320 polarimeter (1 dcm
cell). Thin layer chromatography (TLC) was carried out on Baker Si
250F plates. Visualization was accomplished with either iodine
vapor, UV exposure or treatment with phosphomolybdic acid (PMA).
Preparative TLC was carried out on Analtech uniplates Silica Gel GF
2000 microns. Flash chromatography was carried out on Baker Silica
Gel 40 M (SiO.sub.2). Elemental Analyses were performed by Atlantic
Microlab, Atlanta, Ga. A Beckman 1801 Scintillation Counter was
used for scintillation spectrometry. 0.1% Bovine Serum Albumin and
(-)-Cocaine were purchased from Sigma Chemicals. All reactions were
conducted under an atmosphere of dry nitrogen.
[0086] [.sup.3H]WIN 35,428 and
2.beta.-carbomethoxy-3.beta.-(4-fluorophenyl)-N-[.sup.3H]methyltropane
(79.4-87.0 Ci/mmol), and [.sup.3H]citalopram (86.8 Ci/mmol) were
purchased from DuPont-New England Nuclear (Boston, Mass.). TEA is
triethylamine. (-)-Cocaine hydrochloride for the pharmacological
studies was donated by the National Institute on Drug Abuse [NIDA].
Fluoxetine was donated by E. Lilly & Co. HPLC analyses were
carried out on a Waters 510 system with detection at 254 nm on a
Waters 8 mm, C-18, 10 m reverse phase column. Pd.sub.2dba.sub.3 is
trisdibenzylideneacetone dipalladium, TFA is trifluoroacetic acid,
THF is tetrahydrofuran, EtOAc is ethyl acetate.
EXAMPLE 1
(1R)-2-(Methoxycarbonyl)-3-[[(trifluoromethyl)sulfonyl]oxy]trop-2-ene
(Compound 2, FIG. 2)
[0087] (1R)-(-)-2-Methoxycarbonyl-3-tropinone, 1 {Meltzer et al.,
J. Med. Chem, 1994, 37, 2001} (1 g, 5.07 mmol) was dissolved in
anhydrous THF (20 mL) and the resulting solution cooled to
-78.degree. C. A solution of sodium bistrimethylsilylamide (1 M,
5.58 mL, 5.58 mmol) was then added to the solution slowly. After 30
min, N-phenyltrifluoromethane sulfonamide (1.94 g, 5.43 mmol) was
added. The resulting solution stirred for a further 45 min at
-78.degree. C. and then allowed to attain room temperature and
stirred at room temperature for 2 h. All solvent was evaporated and
the residue pumped to dryness. Column chromatography was performed
on the residue (SiO.sub.2 60 g; 2%-16% methanol in ethyl acetate)
and gave 1.62 g (97%) of a yellow oil which crystallized on
standing.
[0088] R.sub.f 0.65 (10% MeOH/EtOAc). .sup.1H-NMR (CDCl.sub.3)
.delta. 1.58 (m, 1H), 1.97 (m, 2H), 2.1-2.2 (m, 2H), 2.39 (s, 3H).
2.84 (dd, J=18, 4 Hz, 1H), 3.42 (t, J=6 Hz, 1 H), 3.8 (s, 3H), 3.92
(d, J=5 Hz, 1H).
EXAMPLE 2
(1R)-N-Methyl-2-methoxycarbonyl-3-(3,4-dichlorophenyl)-8-azabicyclo[3.2.1]-
oct-2-ene (Compound 3, FIG. 2)
[0089] (1R)-2-Methoxycarbonyl-3-[[(trifluoromethyl)sulfonyl]oxy]
tropene 2 (620 mg, 1.88 mmol), LiCl (171 mg, 4.03 mmol),
Pd.sub.2dba.sub.3 (69 mg, 0.075 mmol), aq. Na.sub.2CO.sub.3 (2.0 M,
2 mL), diethoxymethane (6.2 mL) were all charged to a flask and
stirred vigorously. To this solution was added 3,4-dichlorophenyl
boronic acid (474 mg, 2.49 mmol). The reaction was then brought to
reflux for 2 h and filtered through celite. The cake was washed
with ether and the organic solution was washed with concentrated
NH.sub.4OH. The washed solvent was dried with K.sub.2CO.sub.3,
filtered, and evaporated. The residue was charged to a column
(SiO.sub.2, 60 g, eluted with 5-6% Et.sub.3N/EtOAc) and gave 512 mg
(83%) of a yellow oil which solidified upon standing.
[0090] R.sub.f 0.56 (10% Et.sub.3N/EtOAc). IR (KBr) 2941, 1724,
1460, 1418, 1333, 1250, 1212, 1124 cm.sup.-1. .sup.1H-NMR
(CDCl.sub.3) .delta. 1.61 (m, 1H), 1.9-2.05 (m, 2H), 2.1-2.3 (m,
2H), 2.43 (s, 3H), 2.76 (dd, J=19, 4.7 Hz, 1H), 3.36 (t, J=4.9 Hz,
1H), 3.52 (s, 3H), 3.86 (d, J=5.5 Hz, 1H), 6.96 (dd, J=8.3, 1.9 Hz,
1H), 7.2 (d, J=2.2 Hz, 1H), 7.37 (d, J=8.2 Hz, 1H). Elemental
analysis: calculated C, 58.91; H, 5.25; N, 4.29; found C, 58.84; H,
5.24; N, 4.24.
EXAMPLE 3
(1R)-N-Methyl-2.beta.-methoxycarbonyl-3.beta.-(3,4-dichlorophenyl)-8-azabi-
cyclo[3.2.1]octane (Compound 4 (R=3,4-Cl.sub.2), FIG. 2), and
(1R)-N-Methyl-2.beta.-methoxycarbonyl-3.alpha.-(3,4-dichlorophenyl)-8-azab-
icyclo[3.2.1]octane (Compound 12 (R=3,4-Cl.sub.2), FIG. 2)
[0091] To
(1R)-N-Methyl-2-methoxycarbonyl-3-(3,4-dichlorophenyl)-8-azabic-
yclo[3.2.1]oct-2-ene, 3 (4 g, 12.3 mmol) in THF (43 mL) at
-78.degree. C. was added SmI.sub.2 solution (0.1 M in THF, 400 mL,
40 mmol) dropwise. After 30 min at -78.degree. C., MeOH (140 mL)
was added and the resulting solution stirred at -78.degree. C. for
a further 1 h. The reaction was then quenched with TFA (28 mL) and
water (285 mL), the cold bath was removed and the solution allowed
to attain room temperature. The reaction was then made basic with
NH.sub.4OH and diluted with ether and filtered through celite. The
filter cake was washed with ether and all the organic phases were
combined and washed with a sodium thiosulfate solution and then a
brine solution. After drying with Na.sub.2SO.sub.4 the solution was
filtered and concentrated and gave 3.8 g of the crude products. The
title compounds 4 and 12 were isolated by column chromatography
(SiO.sub.2, 10 g; 2.5% EtOH in CHCl.sub.3). Compound 12 was
isolated as colorless crystals (1.15 g, 29%).
[0092] Mp. 89-91.degree. C. R.sub.f 0.64 (1% NH.sub.4OH/EtOAc).
.sup.1H-NMR (CDCl.sub.3) .delta. 1.28 (ddd, J=1.6, 10.4, 14, 1H),
1.4-1.6 (m, 2H), 2.23 (s, 3H), 2.05-2.3 (m, 2H), 2.35-2.5 (m, 2H),
3.2-3.47 (m, 3H), 3.59 (s, 3H), 7.04 (dd, J=2.2, 8.2 Hz, 1H), 7.27
(d, J=2.2 Hz, 1H), 7.30 (d, J=8.2 Hz, 1H). Elemental analysis:
calculated (0.1 C.sub.6H.sub.14) C, 59.18; H, 6.10; N, 4.16; Cl,
21.05; found C, 59.11; H, 5.90; N, 4.08; Cl, 21.01.
[0093] Compound 4 (R=3,4-Cl.sub.2) (Meltzer et al., J. Med. Chem.,
1993, 36, 855-862) was isolated as a yellow solid (1.04 g,
26%).
[0094] Mp. 82.5-83.5.degree. C. R.sub.f 0.43
(IPA/Et.sub.2O/pentane; 3/30/67); .sup.1H NMR (400 MHz, CDCl.sub.3)
.delta. 1.6-1.7 (m, 3H), 2.0-2.1 (m, 2H), 2.21 (s, 3H), 2.50 (ddd,
1H, H-4), 2.86 (m, 1H), 2.92 (m, 1H), 3.33 (m, 1H, H-5), 3.52 (s,
3H), 3.55 (m, 1H), 7.07-7.32 (m, 3H).
[.alpha.].sub.D.sup.21-27.0.degree. (c=1, CH.sub.3OH). Anal.
(C.sub.16H.sub.19NO.sub.2Cl.sub.2) C, H, N, Cl.
EXAMPLE 4
(1R)-N-Methyl-2.beta.-methoxycarbonyl-3.beta.-(4-fluorophenyl)-8-azabicycl-
o[3.2.1]octane (Compound 4 (R=4-F), FIG. 2) and
(1R)-N-Methyl-2.beta.-methoxycarbonyl-3.alpha.-(4-fluorophenyl)-8-azabicyc-
lo[3.2.1]octane (Compound 12 (R=4-F), FIG. 2; O-1204)
[0095] Using the same general procedure as described above for
Example 3, except substituting
(1R)-N-Methyl-2-methoxycarbonyl-3-(4-fluorophenyl)-8-azabicyclo[3.2.1]oct-
-2-ene, 3, compounds 4 and 12 (R=4-F) were obtained.
[0096] Compound 4 (R=4-F): white solid; Mp 93-94.degree. C.;
[0097] R.sub.f 0.42 (i-PrNH.sub.2:Et.sub.2O:pentane::5:30:65);
.sup.1H-NMR (400 MHz, CDCl.sub.3) .delta. 1.57-1.75 (m, 3H),
2.0-2.2 (m, 2H), 2.23 (s, 3H), 2.54 (ddd, 1H), 2.84 (t, 1H), 2.95
(ddd 1H, J=5.3, 12.7 Hz), 3.36 (m, 1H), 3.50 (s, 3H), 3.55 (m, 1H),
6.9-7.25 (m, 4H). [.alpha.].sub.D.sup.21-45.6.degree. (c=1,
CH.sub.3OH). Anal. (C.sub.16H.sub.20NO.sub.2F) C, H, N.
[0098] Compound 12 (R=4-F): colorless oil;
[0099] R.sub.f 0.5 (10% MeOH in EtOAc); Elemental analysis:
calculated C, 69.29; H, 7.27; N, 5.05; found C, 69.35; H, 7.26; N,
5.00; IR 2900, 1750, 1500 cm-1, .sup.1H-NMR (100 MHz, CDCl.sub.3)
.delta. 1.1-1.8 (m, 4H), 1.9-2.6 (m, 3H), 2.25 (s, 3H), 3.1-3.7 (m,
3H), 3.59 (s, 3H), 6.8-7.3 (m, 4H).
EXAMPLE 5
2.beta.-Carboxy-3.beta.-(4-fluorophenyl)tropane (Compound 5
(R=4-F), FIG. 3)
[0100] 2.beta.-Methoxycarbonyl-3.beta.-(4-fluorophenyl)tropane (WIN
35,428), 4 (1.25 g, 4.54 mmol) was refluxed for 24 h in a 1:1
dioxane-water (80 mL) solution. The solvent was removed in vacuo
and the residue was almost completely dissolved in CHCl.sub.3 (275
mL). Remaining undissolved solid was filtered off, toluene (30 mL)
was added, and the solution was reduced in vacuo by approximately
75%. After cooling the resulting white suspension in the freezer
for 2 h, the white solid was removed by filtration and was washed
with cold 1:1 CHCl.sub.3-toluene. The solid was pumped dry to yield
the product 5 as a white solid (1.11 g, 95%).
[0101] .sup.1H-NMR (CDCl.sub.3) .delta. 1.7-1.8 (m, 1H), 1.94 (dd,
J=9 Hz, 2H) 2.24-2.34 (m, 2H), 2.25 (m, 3H), 2.57 (ddd, J=13.7 Hz,
1H), 2.62-2.68 (m, 1H), 3.16 (ddd, J=13 Hz, 1H), 3.5-3.6 (m, 2H),
6.8 (m, 2H), 7.18-7.24 (m, 2H).
EXAMPLE 6
2.beta.-Carboxy-3.beta.-(3,4-dichlorophenyl)tropane (Compound 5
(R=3,4-Cl.sub.2), FIG. 3)
[0102] 2.beta.-Methoxycarbonyl-3.beta.-(3,4-dichlorophenyl)tropane,
4 (1.14 g, 3.47 mmol) was dissolved in THF:MeOH (1:1; 46 mL) to
which was added a solution of LiOH.H.sub.2O (153 mg) in water (11
mL). The solution was heated to reflux for 24 h, cooled to
0.degree. C. and neutralized (pH=7) with conc. HCl. Silica (1.75 g)
was added directly to the solution and solvent was removed in
vacuo. The material was purified by flash chromatography column
(eluent 20% MeOH/CHCl.sub.3 (900 mL) followed by 30%
MeOH/CHCl.sub.3 (2 L)). Fractions containing the product were
collected and combined, evaporated in vacuo and the residue was
dried at high vacuum. The product was obtained (310 mg; 28%).
[0103] R.sub.f 0.08 (30% MeOH/CHCl.sub.3); .sup.1H-NMR (CDCl.sub.3)
.delta. 1.8-1.9 (m, 1H), 2.1-2.2 (m, 2H) 2.3-2.5 (m, 2H), 2.6-2.9
(m, 2H), 2.78 (s, 3H), 3.3-3.4 (m, 1H), 3.93 (m, 2H), 7.25 (dd,
J=8.2 Hz, 2.2 Hz, 1H), 7.41 (d, J=8.2 Hz, 1H), 7.48 (d, J=2.2 Hz,
1H).
EXAMPLE 7
2.beta.- Methoxymethylcarbamoyl-3.beta.-(4-fluorophenyl)tropane
(Compound 6 (R=4-F), FIG. 3)
[0104] To a stirred suspension of the acid 5 (1.1 g) in anhydrous
CH.sub.2Cl.sub.2 (80 mL) containing DMF (50 .mu.L) was added oxalyl
chloride (1 mL, 11.4 mmol) dropwise resulting in copious bubbling
and dissolution of the suspension. The reaction was allowed to stir
for 45 min during which time the solution became yellow. The
solution was then reduced in vacuo and pumped at high vacuum
overnight, care being taken to bleed nitrogen into the evacuated
flask when transferring from the rotary to the pump.
[0105] To the acid chloride dissolved in CH.sub.2Cl.sub.2 (80 mL)
was added (MeO)MeNH HCl (450 mg; dried immediately prior to use
over P.sub.2O.sub.5 under high vacuum followed immediately by
pyridine (1.1 mL). The reaction was allowed to stir for 1 h and was
partitioned across CHCl.sub.3 (20 mL) and 2M Na.sub.2CO.sub.3 (20
mL). The aqueous layer was extracted CHCl.sub.3 (2.times.10 mL) and
the combined organic extracts were dried over Na.sub.2SO.sub.4,
filtered and reduced in vacuo to yield 1.09 g of a yellow solid.
The crude product was dissolved in CH.sub.2Cl.sub.2 and purified by
flash chromatography (SiO.sub.2, 43 g; 20% hexanes/EtOAc, 5%
Et.sub.3N). Product containing fractions were combined and
concentrated to yield a light yellow solid 6 (960 mg; 75%).
[0106] Mp. 120.1-122.5.degree. C.; R.sub.f 0.14 (25% hexanes/EtOAc,
5% TEA); Elemental analysis: calculated C, 66.65; H, 7.57; N, 9.14;
found C, 66.78; H, 7.63; N, 9.01; IR (KBr) 2900, 1680, 1500
cm.sup.-1; .sup.1H-NMR (CDCl.sub.3) .delta. 1.5-1.8 (m, 3H),
2.0-2.3 (m, 2H), 2.24 (m, 3H), 2.79 (ddd, 1H), 3.0 (m, 1H), 3.05
(s, 3H), 3.1 (m, 1H), 3.4 (m, 1H), 3.47 (m, 1H), 3.57 (s, 3H), 6.9
(m, 2H), 7.25 (m, 2H).
EXAMPLE 8
2.beta.-Methoxymethylcarbamoyl-3.beta.-(3,4-dichlorophenyl)tropane
(Compound 6 (R=3,4-Cl.sub.2), FIG. 3)
[0107] To a stirred solution of
2.beta.-carboxy-3.beta.-(3,4-dichlorophenyl)tropane, 5 (300 mg,
9.55 mmol) in anh. CH.sub.2Cl.sub.2 (30 mL) was added anh. DMF (40
.mu.L) and oxalyl chloride (450 .mu.L) dropwise. The solution was
stirred at room temperature for 40 min. The solvent was removed in
vacuo and the residue dried at high vacuum overnight.
[0108] To the dry residue was added methoxy methylamine
hydrochloride (103 mg, 1.05 mmol). The flask was flushed with
nitrogen and CH.sub.2Cl.sub.2 (30 mL) was added by cannula followed
immediately by pyridine (400 .mu.L). The reaction was stirred for
2.5 h. The resultant mixture was partitioned between
CH.sub.2Cl.sub.2 (40 mL) and 1M Na.sub.2CO.sub.3 (25 mL). The
aqueous layer was extracted with CHCl.sub.3 and the combined
organic extracts were dried and concentrated to yield a yellow
solid (298 mg). The solid was purified on a chromatography column
(eluent 50% EtOAc/hexane/5% Et.sub.3N). Like fractions were
combined, solvent removed and the product dried at high vacuum to
yield Compound 6 (39 mg; 11%).
[0109] R.sub.f 0.1 (50% EtOAc/hexane/5% Et.sub.3N); .sup.1H-NMR
(CDCl.sub.3) .delta. 1.5-1.74 (m, 3H), 2.0-2.3 (m, 2H), 2.21 (m,
3H), 2.72 (ddd, 1H), 2.93 (m, 1H), 3.04 (s, 3H), 3.13 (m, 1H), 3.38
(m, 1H), 3.5 (m, 1H), 3.62 (s, 3H), 7.13 (dd, 1H), 7.30 (d, 1H),
7.31 (d, 1H).
EXAMPLE 9
2.beta.-(1-Propanoyl)-3.beta.-(4-fluorophenyl)tropane (Compound 7
(R=4-F), FIG. 3)
[0110] A solution of
2.beta.-(N-methoxy-N-methylcarbamoyl)-3.beta.-(4-fluorophenyl)-tropane,
6 (823 mg, 2.7 mmol) in THF (10 mL) was cooled to 0.degree. C. and
EtMgBr/Et.sub.2O (3M; 3 mL) was added dropwise over 4 min. The
reaction was warmed to room temperature for 30 min and then heated
to 65.degree. C. for 45 min. The mixture was cooled to 0.degree. C.
and quenched by addition of ethereal HCl (3M). The resulting cloudy
solution was basified with 2M Na.sub.2CO.sub.3. Ether (5 mL) was
added and the layers separated and the aqueous layer washed with
Et.sub.2O (1.times.10 mL) and CHCl.sub.3 (2.times.10 mL). The
combined organic extracts were dried over Na.sub.2SO.sub.4,
filtered and concentrated. The product was purified by flash column
chromatography (eluent 25% EtOAc/hexanes/5% TEA) to provide 7 (485
mg, 65%).
[0111] R.sub.f 0.3 (20% EtOAc/hexanes/5% Et.sub.3N); mp
118-119.5.degree. C.; Elemental analysis: calculated C, 74.15; H,
8.05; N, 5.09; found C, 74.05; H, 8.09; N, 5.00. IR (KBr) 2900,
1710, 1500. 1250 cm.sup.-1; .sup.1H-NMR (CDCl.sub.3) .delta. 0.85
(t, 3H), 1.5-1.8 (m, 4H), 2.0-2.4 (m, 3H), 2.23 (m, 3H), 2.5-2.6
(m, 1H), 2.9-3.0 (m, 2H), 3.36 (m, 1H), 3.48 (m, 1H), 6.69 (m, 2H),
7.17 (m, 2H).
EXAMPLE 10
2.beta.-(1-Propanoyl)-3.beta.-(3,4-dichlorophenyl)tropane (Compound
7 (R=3,4-Cl.sub.2), FIG. 3)
[0112] A solution of
2.beta.-methoxymethylcarbamoyl-3.beta.-(3,4-dichlorophenyl), 6 (168
mg, 0.47 mmol) in THF (30 mL) was cooled to 0.degree. C. and
EtMgBr/Et.sub.2O (3M; 1 mL) was added dropwise over 3 min. The
reaction was warmed to room temperature for 60 min and then heated
to reflux for 10 min. The mixture was cooled to room temperature
and quenched by addition to ethereal HCl (3M, 20 mL). The resulting
cloudy solution was basified with saturated aqueous NaHCO.sub.3 and
brought to pH=10 by addition of Na.sub.2CO.sub.3. Ether was added
and the layers separated and the aqueous layer washed with
Et.sub.2O (3.times.25 mL). The combined organic extracts were dried
over Na.sub.2SO.sub.4, filtered and concentrated. The product (166
mg) was purified by flash column chromatography (eluent 30%
EtOAc/hexanes/5% Et.sub.3N) to provide 7 (108 mg, 71%).
[0113] R.sub.f 0.32 (50% EtOAc/hexanes/5% Et.sub.3N); .sup.1H-NMR
(CDCl.sub.3) .delta. 0.9 (t, 3H), 1.5-1.8 (m, 4H), 2.0-2.3 (m, 3H),
2.2 (m, 3H), 2.35-2.55 (m, 2H), 2.82-2.92 (m, 1H), 2.96 (m, 1H),
3.34 (m, 1H), 3.54 (m, 1H), 7.75 (dd, 1H), 7.27 (d, 1H), 7.29 (d,
1H).
EXAMPLE 11
2.beta.-(1-Propanoyl)-3.beta.-(4-fluorophenyl)nortropane (Compound
8 (R=4-F), FIG. 3)
[0114] 2.beta.-(1-Propanoyl)-3.beta.-(4-fluorophenyl)tropane, 7
(335 mg) was combined with 1-chloroethyl chloroformate (5 mL) and
the solution was heated to reflux for 5 h. The excess chloroformate
was removed in vacuo and the residue was refluxed in methanol for
1.5 h. The methanol was removed in vacuo and the residue was
dissolved in CHCl.sub.3 (15 mL) and shaken with 2M
Na.sub.2CO.sub.3. The aqueous layer was extracted CHCl.sub.3
(2.times.15 mL) and the combined organic extracts were dried
(Na.sub.2SO.sub.4), filtered and concentrated to yield 369 mg. This
residue was chromatographed (eluent: 100 mL EtOAc, 150 mL 5%
Et.sub.3N/EtOAc, 100 mL 10% Et.sub.3N/EtOAc, 300 mL 20%
Et.sub.3N/EtOAc, and 300 mL 30% Et.sub.3N/EtOAc). Like fractions
were combined to yield 8 (104 mg, 33%).
[0115] R.sub.f 0.36 (10% Et.sub.3N/EtOAc); .sup.1H-NMR (CDCl.sub.3)
.delta. 0.70 (t, 3H), 1.4-1.8 (m, 5H), 1.9-2.3 (m, 3H), 2.4 (m,
1H), 2.88(m, 1H), 3.18 (m, 1H), 3.56 (m, 1H), 3.70 (m, 1H), 6.94
(m, 2H), 7.10 (m, 2H).
EXAMPLE 12
2.beta.-(1-Propanoyl)-3.beta.-(3,4-dichlorophenyl)nortropane
(Compound 8 (R=3,4-Cl.sub.2), FIG. 3)
[0116] 2.beta.-(1-Propanoyl)-3.beta.-(3,4-dichlorophenyl)tropane 7
(107 mg, 0.32 mmol) was combined with 1-chloroethyl chloroformate
(2 mL) and the solution was heated to reflux for 5 h. The excess
chloroformate was removed in vacuo and the residue was refluxed in
methanol for 45 min. The methanol was removed in vacuo and the
residue was dissolved in CH.sub.2Cl.sub.2 and shaken with
NaHCO.sub.3/Na.sub.2CO.sub.3 (pH=9). The aqueous layer was
extracted CH.sub.2Cl.sub.2 (4.times.10 mL) and the combined organic
extracts were dried (Na.sub.2SO.sub.4), filtered and concentrated
to yield 127 mg. This residue was chromatographed (1.times.100 mL
5% Et.sub.3N/EtOAc; 3.times.100 mL 10% Et.sub.3N/EtOAc). Like
fractions were combined to yield 8 (30 mg, 29%).
[0117] .sup.1H-NMR (CDCl.sub.3) .delta. 0.76 (t, 3H), 1.4-2.5 (m,
9H), 2.92 (m, 1H), 3.09-3.2 (m, 1H), 3.6 (m, 1H), 3.72 (m, 1H), 7.0
(m, 1H), 7.25 (d, 1H), 7.32 (d, 1H).
EXAMPLE 13
N-[2-(3'-N'-Propyl-(1''R)-3''.beta.-(4-fluorophenyl)tropane-2''.beta.-(1-p-
ropanoyl))((2-((triphenylmethyl)
thio)ethyl)amino)acetyl]-S-(triphenyl)-2-aminoethanethiol (Compound
9 (R=4-F), FIG. 3 (O-1507)
[0118] 2.beta.-(1-Propanoyl)-3.beta.-(4-fluorophenyl)nortropane 8
(27 mg) was combined with MAMA'-Cl (86 mg), KI (34 mg, 2.0 eq.),
and NaHCO.sub.3 (43 mg, 5 eq.) in anhydrous MeCN (4 mL) and brought
to reflux for 4 h. The solvent was removed under vacuum and the
residue was partitioned between CHCl.sub.3 and saturated aqueous
NaHCO.sub.3. The aqueous layer was extracted CHCl.sub.3 (2.times.5
mL) and the combined organic extracts were dried
(Na.sub.2SO.sub.4), filtered and concentrated to yield a brown
foam. The foam was applied to a chromatography column (10 g silica;
40% EtOAc in hexanes/1% TEA). Fractions containing the product were
combined and concentrated to yield 9 as a light foam (47 mg,
46%).
[0119] R.sub.f 0.09 (60% EtOAc/hexanes, 1% Et.sub.3N). Elemental
analysis: calculated C, 72.15; H, 6.34; N, 3.96; found C, 71.99; H,
6.41; N, 3.92. .sup.1H-NMR (CDCl.sub.3) .delta. 0.77 (t, 3H),
1.2-3.1 (m, 29H), 3.3-3.5 (m, 2H), 6.9-7.0 (m, 2H), 7.1-7.6 (m, 32
H).
EXAMPLE 14
N-[2-(3'-N'-Propyl-(1''R)-3''.beta.-(3,4-dichlorophenyl)tropane-2''.beta.--
(1-propanoyl))((2-((triphenylmethyl)
thio)ethyl)amino)acetyl]-S-(triphenyl)-2-aminoethanethiol (Compound
9 (R=3,4-Cl.sub.2), FIG. 3
[0120] 2.beta.-(1-Propanoyl)-3.beta.-(3,4-dichlorophenyl)nortropane
8 (30 mg, 0.096 mmol) was combined with MAMA'-Cl (87 mg, 0.115
mmol, 1.2 eq.), KI (32 mg, 0.19 mmol, 2.0 eq.), and NaHCO.sub.3 (40
mg, 0.48 mmol, 5 eq.) in anhydrous MeCN (4 mL) and brought to
reflux for 4 h then cooled to room temperature and allowed to stir
overnight. The solvent was removed under vacuum and the residue was
partitioned between CH.sub.2Cl.sub.2 (15 mL) and saturated aqueous
NaHCO.sub.3 (15 mL). The aqueous layer was extracted with
CH.sub.2Cl.sub.2 (3.times.10 mL) and the combined organic extracts
were dried (Na.sub.2SO.sub.4), filtered and concentrated to yield a
yellow oil (115 mg). The oil was applied to a chromatography column
(10 g SiO.sub.2; 30% EtOAc in hexanes/1% Et.sub.3N). Fractions
containing the product were combined and concentrated to yield 9 as
a light foam (41 mg, 41%).
[0121] R.sub.f 0.14 (60% EtOAc/hexanes, 1% Et.sub.3N). .sup.1H-NMR
(CDCl.sub.3) .delta. 0.81 (t, 3H), 1.2-3.1 (m, 29H), 3.34 (m, 1H),
3.52 (m, 1H), 7.0-7.6 (m, 33H).
EXAMPLE 15
N-[(2-((3'-N'-Propyl-(1''R)-3''.beta.-(4-fluorophenyl)tropane-2''.beta.-1--
propanoyl)(2-mercaptoethyl)amino)acetyl)-2-aminoethanethiolato]rhenium
(V) oxide (Compound 10 (R=4-F), FIG. 3 (O-1508R)
[0122]
N-[2-(3'-N'-Propyl-(1''R)-3''.beta.-(4-fluorophenyl)tropane-2''.be-
ta.-(1-propanoyl))((2-((triphenylmethyl)
thio)ethyl)amino)acetyl]-S-(triphenyl)-2-aminoethanethiol (22 mg,
0.023 mmol) was dissolved in boiling EtOH (abs. 4 mL) and
SnCl.sub.2 (8.5 mg, in 0.5 mL of 0.05M HCl). The reaction was
maintained at reflux for a further 6 h and silica was added and the
solvents removed by evaporation. The silica adsorbed product was
applied to a silica column (3 g eluent: 30% EtOAc in hexanes with
5% Et.sub.3N). The compound was obtained as a foam (6.7 mg,
44%).
[0123] R.sub.f 0.07 (60% EtOAc in hexanes+NH.sub.4OH (0.5%))
Accurate Mass calc for C.sub.25H.sub.35FN.sub.3O.sub.3ReS.sub.2:
695.172; found 695.162. .sup.1H-NMR (CDCl.sub.3) .delta. 0.7-0.9
(2t, 2H), 1.4-4.1 (m, 26H), 4.5-4.6 (m, 1H), 4.73 (d, J=16.5 Hz,
0.5H) 4.87 (d, J=16.5 Hz, 0.5H), 6.93 (m, 2H), 7.14 (m, 2H).
EXAMPLE 16
2.beta.-(Carboxylic acid)-3.alpha.-(4-fluorophenyl)tropane
(Compound 13 (R=4-F), FIG. 4)
[0124] A solution of
2.beta.-methoxycarbonyl-3.alpha.-(4-fluorophenyl)-tropane 12 (1.0
g, 3.6 mmol) was refluxed for 24 h in 80 mL dioxane water (1:1).
The solvent was removed in vacuo and the brown solid was purified
by column chromatography (eluent 30% MeOH/CHCl.sub.3). The product
(890 mg, 94%) was obtained as a white foam.
[0125] R.sub.f 0.43 (30% MeOH/CHCl.sub.3); NMR .sup.1H-NMR
(CDCl.sub.3) .delta. 1.48-1.62 (m, 1H), 1.75-2.1 (m, 3H), 2.36-2.46
(m, 1H), 2.60 (s, 3H), 2.7-2.82 (m, 1H), 3.22 (brs, 1H), 3.5-3.62
(m, 2H), 3.76-3.84 (m, 1H), 6.9-7.1 (m, 2H), 7.4-7.5 (m, 2H).
EXAMPLE 17
2.beta.-(Carboxylic acid) 3.alpha.-(3,4-dichlorophenyl)tropane
(Compound 13 (R=3,4-Cl.sub.2), FIG. 4)
[0126] A solution of
2.beta.-methoxycarbonyl-3.alpha.-(3,4-dichlorophenyl)tropane 12
(750 mg, 2.29 mmol) and LiOH (335 mg, 8.0 mmol) was brought to
reflux for 4 h in water (10 mL) and THF:MeOH (33 mL; 1:1). The
reaction was neutralized by dropwise addition of conc. HCl. The
solvent was removed in vacuo and the product purified by column
chromatography-(eluent 15% MeOH/CHCl.sub.3). The product 13 (521
mg, 72%) was obtained as a solid.
[0127] R.sub.f 0.15 (30% MeOH/CHCl.sub.3); .sup.1H-NMR (CD.sub.3OD)
.delta. 1.59 (m, 1H), 1.8-1.9 (m, 1H), 2.1-2.2 (m, 2H), 2.6-2.7 (m,
2H), 2.76 (s, 3H), 3.25 (brs, 1H), 3.35 (brs, 1H), 3.85 (m, 1H),
3.93 (m, 1H), 7.5 (d, 1H), 7.53 (dd, 1H), 7.8 (d, 1H).
EXAMPLE 18
2.beta.-Methoxymethylcarbamoyl-3.alpha.-(4-fluorophenyl)tropane
(Compound 14 (R=4-F), FIG. 4) (O-1403)
[0128] To a stirred suspension of Compound 13 (930 mg, 3.6 mmol) in
anhydrous CH.sub.2Cl.sub.2 (80 mL) containing DMF (50 .mu.L) was
added oxalyl chloride (1 mL, 11.4 mmol) dropwise resulting in
copious bubbling and dissolution of the suspension. The reaction
was allowed to stir for 45 min during which time the solution
became yellow. It was then reduced in vacuo and pumped at high
vacuum overnight, care being taken to bleed nitrogen into the
evacuated flask when transferring from the rotary to the pump.
[0129] To the acid chloride dissolved in CH.sub.2Cl.sub.2 (70 mL)
was added (MeO)MeNH.HCl (382 mg, 3.9 mmol) followed immediately by
pyridine (1 mL). The reaction was allowed to stir for 1 h and then
partitioned between CHCl.sub.3 (20 mL) and 2M Na.sub.2CO.sub.3 (20
mL). The aqueous layer was extracted with CHCl.sub.3 (2.times.10
mL) and the combined organic extracts were dried over
Na.sub.2SO.sub.4, filtered and reduced in vacuo to yield a yellow
oil. The oil was dissolved in toluene and concentrated to yield a
yellow solid (653 mg). The crude product was dissolved in a minimum
volume of CHCl.sub.3, and applied to a chromatography column (28 g
SiO.sub.2; eluent 25% hexanes in EtOAc, 5% Et.sub.3N; followed by
25% MeOH in CHCl.sub.3). Product containing fractions were combined
and concentrated to yield the amide (540 mg; 50%).
[0130] R.sub.f 0.25 (25% hexanes/EtOAc, 5% Et.sub.3N); mp.
139.8-141.7.degree. C.; .sup.1H-NMR (CDCl.sub.3) .delta. 1.24 (dd,
1H), 1.4-1.8 (m, 3H), 2.1-2.3 (m, 2H), 2.23 (s, 3H); 2.4-2.6 (m,
1H), 2.7 (brd, 1H), 3.05 (s, 3H), 3.05-3.1 (m, 1H), 3.25-3.5 (m,
2H), 3.41 (s, 3H), 6.86-6.96 (m, 2H), 7.13-7.22 (m, 2H); IR (KBr)
2900, 1656, 1500 cm.sup.-1. Elemental analysis: calculated C,
66.65; H, 7.57; N, 9.14; found C, 66.37; H, 7.59; N, 9.0.
EXAMPLE 19
2.beta.-Methoxymethylcarbamoyl-3.alpha.-(3,4-dichlorophenyl)tropane
(Compound 14 (R=3,4-Cl.sub.2), FIG. 4)
[0131] To a stirred suspension of the acid 13 (210 mg, 0.67 mmol)
in anhydrous CH.sub.2Cl.sub.2 (10 mL) containing DMF (30 .mu.L) was
added oxalyl chloride (0.3 mL, 2.0 mmol) dropwise. The reaction was
allowed to stir for 1 h and then reduced in vacuo and pumped at
high vacuum overnight, care being taken to bleed nitrogen into the
evacuated flask when transferring from the rotary to the pump.
[0132] To the acid chloride in CH.sub.2Cl.sub.2 (10 mL) was added
(MeO)MeNH. HCl (72 mg, 0.74 mmol) followed immediately by pyridine
(0.3 mL). The reaction was allowed to stir for 1.5 h and was
partitioned between CH.sub.2Cl.sub.2 (10 mL) and 1M
Na.sub.2CO.sub.3 solution (5 mL). The aqueous layer was extracted
with CH.sub.2Cl.sub.2 (10 mL) and the combined organic extracts
were dried over Na.sub.2SO.sub.4, filtered and reduced in vacuo to
yield a yellow solid. The solid was purified by column
chromatography (14 g SiO.sub.2, eluent 25% hexanes/EtOAc, 5%
Et.sub.3N). Product containing fractions were combined and
concentrated to yield the amide (115 mg; 48%).
[0133] R.sub.f 0.14 (40% hexanes/EtOAc, 5% Et.sub.3N); .sup.1H-NMR
(CDCl.sub.3) .delta. 1.17 (ddd, 1H), 1.48 (ddd, 1H), 1.63 (ddd,
1H), 2.1-2.34 (m, 2H), 2.21 (s, 3H), 2.42-2.54 (m, 1H), 2.65 (brd,
1H), 3.06 (s, 3H), 3.08 (brs, 1H), 3.22-3.32 (m, 1H), 3.34-3.46 (m,
1H), 3.48 (s, 3H), 7.05 (dd, 1H), 7.25 (d, 1H), 7.27 (d, 1H).
EXAMPLE 20
2.beta.-(1-Propanoyl)-3.alpha.-(4-fluorophenyl)-tropane (Compound
15 (R=4-F), FIG. 4) (O-1369)
[0134] A 250 mL round bottom flask containing the
2.beta.-methoxymethylcarbamoyl-3.alpha.-(4-fluorophenyl)tropane 14
(471 mg) was flushed with nitrogen and charged with anhydrous THF
(70 mL). At room temperature, EtMgBr/Et.sub.2O (3.0 mL; 3.0M) was
added dropwise over 3 min. The reaction was stirred at room
temperature for 30 min and was then heated to 65.degree. C. for 1 h
at which point no starting material was observed by TLC (TLC sample
was prepared by adding an aliquot of the reaction to ethereal HCl,
and basifying with 2M Na.sub.2CO.sub.3; R.sub.f (product) 0.42;
R.sub.f (starting material) 0.13 (20% EtOAc/hexanes, 5% Et.sub.3N).
The reaction was cooled in an ice bath and quenched by slow
addition of ethereal HCl. The cloudy solution was basified with 2M
Na.sub.2CO.sub.3 and diluted with ether (25 mL). The layers were
separated and the aqueous layer was extracted with ether
(1.times.10 mL) and CHCl.sub.3 (2.times.20 mL). The combined
organic extracts were dried (Na.sub.2SO.sub.4), filtered and
reduced in vacuo to yield the crude residue (484 mg). This residue
was then chromatographed (25 g SiO.sub.2; eluent 25% EtOAc/hexanes,
5% Et.sub.3N). Fractions containing the product were combined and
concentrated to yield 15 (300 mg, 70%);
[0135] Mp. 60.5-61.3.degree. C.; R.sub.f 0.49 (33% EtOAc/hexanes;
5% Et.sub.3N); .sup.1H-NMR (CDCl.sub.3) .delta. 0.86 (t, 3H), 1.27
(ddd, 1H), 1.4-1.6 (m, 2H), 2.0-2.5 (m, 6H), 2.23 (s, 3H), 3.12
(brd, 1H), 3.2-3.3 (m, 2H), 6.85-7.0 (m, 2H), 7.05-7.15 (m, 2H); IR
(KBr) 2900, 1740, 1500 cm.sup.-1; Elemental analysis: calculated C,
74.15; H, 8.05; N, 5.09; found C, 74.00; H, 8.13; N, 4.98
EXAMPLE 21
2.beta.-(1-Propanoyl)-3.alpha.-(3,4-dichlorophenyl)tropane
(Compound 15 (R=3,4-Cl.sub.2), FIG. 4)
[0136]
2-Methoxymethylcarbamoyl-3.alpha.-(3,4-dichlorophenyl)tropane, 14
(105 mg, 0.29 mmol) was flushed with nitrogen and charged with
anhydrous THF (15 mL). At room temperature, EtMgBr/Et.sub.2O (0.8
mL; 3.0M) was added dropwise over 3 min. The reaction was stirred
at room temperature for 1 h and was then heated to 55.degree. C.
for 30 min at which point no starting material was observed by TLC.
The reaction was cooled in an ice bath and quenched by slow
addition of ethereal HCl. The cloudy solution was basified with 2M
Na.sub.2CO.sub.3 and diluted with ether (15 mL) and water (15 mL).
The layers were separated and the aqueous layer was extracted with
CHCl.sub.3 (2.times.15 mL). The combined organic extracts were
dried (Na.sub.2SO.sub.4), filtered and reduced in vacuo to yield a
residue (95 mg) which was chromatographed (5 g SiO.sub.2, eluent
25% EtOAc in hexanes, 5% Et.sub.3N). Fractions containing the
product were combined and concentrated to yield 15 (80 mg,
80%).
[0137] R.sub.f 0.28 (30% EtOAc/hexanes; 5% Et.sub.3N); .sup.1H-NMR
(CDCl.sub.3) 0.93 (t, J=7.4 Hz, 3H), 1.27 (ddd, 1H), 1.42-1.62 (m,
2H), 2.06-2.30 (m, 6H), 2.21 (s, 3H), 3.32-2.52 (m, 3H), 3.14 (brd
1H), 3.2-3.36 (m, 2H), 7.10 (dd, 1H), 7.24 (d, 1H), 7.29 (d,
1H).
EXAMPLE 21a
2.beta.-(1-Propanoyl)-3.alpha.-(3,4-dichlorophenyl)tropane
(Compound 15 (R-3,4-Cl.sub.2), FIG. 4)
[0138] To commercially available ethylmagnesium bromide (1M in THF,
12.6 mL, 12.6 mmol) in a flask equipped with an addition funnel
under nitrogen was added triethylamine (5.0 g, 50.4 mmol). To the
resulting mixture was added drop-wise a solution of compound 12
(R=Cl.sub.2, 750 mg, 2.29 mmol) in benzene (10 mL) at 5-10.degree.
C. over a period of 1 hour. The reaction mixture was then stirred
at 5-10.degree. C. for 5 hours and then treated with 4 M HCl (2.9
mL, 11.6 mmol). The organic layer was washed with water (1.times.50
mL), 5% NaHCO.sub.3 (aq) (1.times.50 mL) and water (2.times.50 mL).
The organic phase was then dried (K.sub.2CO.sub.3), filtered and
the concentrated. The residue was chromatographed (SiO.sub.2. 25%
EtOAc in hexanes with 5% Et.sub.3N) and gave 670 mg (85%) of
compound 15 with the same physical and spectral characteristics as
previously reported (Example 21).
EXAMPLE 22
2.beta.-(1-Propanoyl)-3.alpha.-(4-fluorophenyl)nortropane Compound
16 (R=4-F), FIG. 4) (O-1370
[0139] 2.beta.-(1-Propanoyl)-3.alpha.-(4-fluorophenyl)tropane 15
(O-1369) (556 mg, 2 mmol) and ACE-Cl (7 mL) were combined and
brought to reflux for 4 h. All volatiles were removed by
evaporation and methanol (100 mL) was added to the residue. The
resulting mixture was brought to reflux for 90 min. Volatiles were
removed and the residue was taken up in CHCl.sub.3 and washed with
aq. 2M Na.sub.2CO.sub.3. The aqueous mixture was extracted with
CHCl.sub.3 (2.times.10 mL) and organic fractions combined, dried
(Na.sub.2SO.sub.4), filtered and reduced. The dark brown oil (615
mg) was chromatographed (SiO.sub.2, 20 g; 0-10% Et.sub.3N in EtOAc)
and gave 390 mg (74%) of a yellow oil.
[0140] R.sub.f 0.25 (5% Et.sub.3N in EtOAc); .sup.1H-NMR
(CDCl.sub.3) .delta. 0.86 (t, 3H), 1.27 (ddd, 1H), 1.50-1.72 (m,
2H), 1.8-2.1 (m, 3H), 2.2-2.4 (m, 2H), 2.56 (dd, J=10.7, 1 Hz, 1H),
3.07 (ddd, J=11.3, 10.9, 7.1 Hz, 1H), 3.43 (brd 1H), 3.62 (brt,
1H), 6.89-6.99 (m, 2H), 7.09-7.16 (m, 2H). IR (KBr) 2900, 1711,
1500 cm.sup.-1; Elemental analysis: calculated (0.2 H.sub.2O) C,
72.52; H, 7.77; N, 5.25; found C, 72.65; H, 7.81; N, 5.27
EXAMPLE 23
2.beta.-(1-Propanoyl)-3.alpha.-(3,4-dichlorophenyl)nortropane
(Compound 16 (R=3,4-Cl.sub.2), FIG. 4)
[0141] 2.beta.-(1-Propanoyl)-3-(3,4-dichlorophenyl)tropane, 15 (80
mg, 2.45 mmol) was combined with 1-chloroethyl chloroformate (3 mL)
and the solution was heated to reflux for 5 h. The excess
chloroformate was removed in vacuo and the residue was refluxed in
methanol for 1 h. The methanol was removed in vacuo and the residue
was dissolved in CHCl.sub.3 and washed with 2M Na.sub.2CO.sub.3.
The aqueous layer was extracted with CHCl.sub.3 (2.times.5 mL) and
the combined organic extracts were dried (Na.sub.2SO.sub.4),
filtered and concentrated to yield pure compound (77 mg; 100%).
[0142] R.sub.f 0.3 (5% Et.sub.3N/EtOAc); .sup.1H-NMR (CDCl.sub.3)
.delta. 0.91 (t, 3H), 1.25 (ddd, 1H, 1.52-1.74 (m, 2H), 1.8-2.56
(m, 7H), 3.06-3.2 (m, 1H), 3.45 (d, 1H), 3.62 (brt, 1H), 7.01 (dd,
1H), 7.25 (d, 1H), 7.31 (d, 1H).
EXAMPLE 24
N-[2-(3'-N'-Propyl-(1''R)-3''.alpha.-(4-fluorophenyl)tropane-2''.beta.-(1--
propanoyl)) ((2-((triphenylmethyl)
thio)ethyl)amino)acetyl]-S-(triphenyl)-2-aminoethanethiol (compound
17 (R=4-F), FIG. 4) (O-1506
[0143] 2.beta.-(1-Propanoyl)-3.alpha.-(4-fluorophenyl)nortropane 16
(24 mg, 0.09 mmol), MAMA'-Cl (147 mg, 0.19 mmol), KI (31 mg, 19
mmol) were all dissolved in CH.sub.3CN (3 mL) and brought to reflux
for 3 h. Solvent was removed in vacuo and the residue partitioned
between satd. aq NaHCO.sub.3 and CHCl.sub.3. The aqueous layer was
further extracted with CHCl.sub.3, all organic fractions were
combined and dried (Na.sub.2SO.sub.4), filtered and concentrated.
Column chromatography (20% EtOAc, 79% Hexane, 1% Et.sub.3N) gave
pure product as a golden foam (11.5 mg, 13%).
[0144] R.sub.f 0.15 (50% EtOAc in hexanes+5% Et.sub.3N); Elemental
analysis: calculated (0.1H.sub.2O) C, 75.79; H, 6.87; N, 4.21;
found C, 75.35; H, 6.81; N, 4.13. .sup.1H-NMR (CDCl.sub.3) .delta.
0.85 (t,3H), 1.19 (ddd, 1H), 1.3-1.5 (m, 6H), 1.8-2.6 (m, 14H),
2.85 (brs, 2H), 3.0 (m, 2H), 3.08-3.30 (m, 3H), 6.88-6.96 (m, 2H),
7.06-7.42 (m, 32H).
EXAMPLE 25
N-[2-(3'-N'-Propyl-(1''R)-3''.alpha.-(3,4-dichlorophenyl)tropane-2''.beta.-
-(1'''-propanoyl))((2-((triphenylmethyl)
thio)ethyl)amino)acetyl]-S-(triphenyl)-2-aminoethanethiol (Compound
17 (R=3,4-Cl.sub.2), FIG. 4) (O-1546)
[0145]
2.beta.-(1-Propanoyl)-3.alpha.-(3,4-dichlorophenyl)nortropane 16
(75 mg, 0.24 mmol) was combined with N-(((2-(2-(triphenylmethyl)
thio)ethyl)(N'-3'-chloropropyl)amino)acetyl)-S-(triphenylmethyl)-2-aminoe-
thanethiol (MAMA'-Cl) (218 mg, 0.29 mmol), KI (80 mg, 0.48 mmol),
and NaHCO.sub.3 (101 mg, 1.2 mmol) in anhydrous MeCN (20 mL) and
brought to reflux for 4 h then cooled to room temperature. The
solvent was removed under vacuum and the residue was partitioned
between CH.sub.2Cl.sub.2 (20 mL) and saturated aqueous NaHCO.sub.3
(15 mL). The aqueous layer was extracted with CH.sub.2Cl.sub.2
(1.times.10 mL) and the combined organic extracts were dried
(Na.sub.2SO.sub.4), filtered and concentrated to yield a brown oil.
The oil was applied to a chromatography column (30 g SiO.sub.2;
eluent: 25% EtOAc/hexanes/1% Et.sub.3N). Fractions containing the
product were combined and concentrated to yield 17 (51 mg,
17%).
[0146] R.sub.f 0.3 (60% EtOAc in hexanes, 1% Et.sub.3N); Elemental
analysis: calculated (1.33 H.sub.2O) C, 71.71; H, 6.46; N, 3.98;
found C, 71.85; H, 6.52; N, 3.91. .sup.1H-NMR (CDCl.sub.3) .delta.
0.89 (t, 3H), 1.0-1.7 (m, 8H), 1.8-2.5 (m, 14H), 2.6-3.4 (m, 6H),
6.9-7.6 (m, 33H).
EXAMPLE 26
N-[(2-((3'-N'-Propyl-(1''R)-3''.alpha.-(4-fluorophenyl)tropane-2''.beta.-1-
'''-propanoyl)
(2-mercaptoethyl)amino)acetyl)-2-aminoethanethiolato]rhenium (V)
oxide (Compound 18 (R=4-F), FIG. 4) (O-1505)
[0147] A solution of
N-[2-(3'-N'-Propyl-(1''R)-3''.alpha.-(4-fluorophenyl)tropane-2''.beta.-(1-
'''-propanoyl))((2-((triphenylmethyl)
thio)ethyl)amino)acetyl]-S-(triphenyl)-2-aminoethanethiol, 17 (22
mg) in EtOH (4 mL) was heated to reflux. A solution of SnCl.sub.2
(7.8 mg, 0.04 mmol) in 0.005 M HCl (0.5 mL) was added quickly
followed immediately by NaReO.sub.4 (12.4 mg, 0.04 mmol) in 0.005 M
HCl (0.5 mL). The cloudy solution was refluxed for 6 h and was then
loaded onto 0.5 g silica and pumped overnight. The silica-adsorbed
material was applied to a chromatography column (3 g SiO.sub.2;
eluent 30% EtOAc/hexanes/5% TEA). The purple brown solid obtained
was triturated with pentane and dried at high vacuum overnight to
yield 18 as a foam (9.9 mg; 71%).
[0148] Accurate Mass calc 695.161 (found, 695.172). R.sub.f 0.30
(75% EtOAc in hexane+0.5% NH.sub.4OH); .sup.1H-NMR (CDCl.sub.3)
.delta. 0.75-0.95 (2t, 3H), 1.0-2.0 (m, 8H), 2.0-2.5 (m, 6H),
2.8-3.0 (m, 1H), 3.0-3.5 (m, 7H), 3.6-3.8 (m, 1H), 3.9-4.2 (m, 3H),
4.5-4.65 (m, 1H), 4.73, 5.12 (2d, J=16.7 Hz), 6.9-7.0 (m, 2H),
7.08-7.15 (m, 2H).
EXAMPLE 27
N-[(2-((3'-N'-Propyl-(1''R)-3''.alpha.-(3,4-dichlorophenyl)tropane-2''.bet-
a.-1'''-propanoyl)
(2-mercaptoethyl)amino)acetyl)-2-aminoethanethiolato]rhenium (V)
oxide (Compound 18 (R=3,4-Cl.sub.2), FIG. 4)
[0149] A solution of
N-[2-(3'-N'-Propyl-(1''R)-3''.alpha.-(3,4-dichlorophenyl)tropane-2''.beta-
.-(1-propanoyl)) (2-((triphenylmethyl)
thio)ethyl)amino)acetyl]-S-(triphenyl)-2-aminoethanethiol, 17, (24
mg, 0.024 mmol) in EtOH (5 mL) was heated to reflux. A solution of
SnCl.sub.2 (9 mg, 0.05 mmol) in 0.005 M HCl (0.5 mL) was added
quickly followed immediately by NaReO.sub.4 (14.5 mg, 0.05 mmol) in
0.005 M HCl (0.5 mL). The solution was refluxed for 4 h and was
then loaded onto 0.5 g silica and pumped at high vacuum overnight.
The silica-adsorbed material was applied to a chromatography column
(4 g SiO.sub.2; eluent 30% EtOAc/hexanes/5% Et.sub.3N). The solid
obtained was concentrated to yield 18 (4.6 mg; 27%).
[0150] .sup.1H-NMR (CDCl.sub.3) .delta. 0.8-1.0 (2t, 3H), 1.1-2.5
(m, 14H), 2.8-3.0 (m, 1H), 3.0-3.5 (m, 7H), 3.6-3.8 (m, 1H),
3.9-4.2 (m, 3H), 4.4-4.65 (m, 1H), 4.70, 5.06 (2d, 2.times.J=16.7
Hz, 1H), 7.0 (dd, 1H), 7.3 (d, 1H), 7.34 (d, 1H).
EXAMPLE 28
2.beta.-(Methoxymethylcarbamoyl-3.alpha.-(4-fluorophenyl)nortropane
(Compound 20 (R=4-F), FIG. 5)
[0151] 2.beta.-(1-Propanoyl)-3.alpha.-(4-fluorophenyl)tropane 14
(112 mg, 0.37 mmol) was combined with 1-chloroethyl chloroformate
(2.2 mL) and the solution was heated to reflux for 5 h. The excess
chloroformate was removed in vacuo and the residue was refluxed in
methanol for 45 min. The MeOH was removed in vacuo and the residue
was dissolved in CHCl.sub.3 and NaHCO.sub.3/Na.sub.2CO.sub.3
(pH=9). The aqueous layer was extracted CHCl.sub.3 (3.times.10 mL)
and the combined organic extracts were dried (Na.sub.2SO.sub.4),
filtered and concentrated to yield 104 mg. This residue was
chromatographed (eluent: 10-20% Et.sub.3N in EtOAc). Like fractions
were combined to yield 20 (65 mg, 60%).
[0152] R.sub.f 0.13 (10% Et.sub.3N in EtOAc); .sup.1H-NMR
(CDCl.sub.3) .delta. 1.30 (ddd, 1H), 1.5-1.75 (m, 2H), 1.85-2.15
(m, 2H), 2.2-2.35 (m, 1H), 2.7-2.9 (m, 1H), 3.05 (s, 3H), 3.1-3.22
(m, 1H), 3.38 (s, 3H), 3.4-3.5 (m, 1H), 3.6-3.7 (m, 1H), 6.9-7.0
(m, 2H), 7.13-7.23 (m, 2H).
EXAMPLE 29
N-[2-(3'-N'-Propyl-(1''R)-3''.alpha.-(4-fluorophenyl)tropane-2''.beta.-(me-
thoxymethylcarbamoyl))((2-((triphenylmethyl)thio)ethyl)amino)acetyl]-S-(tr-
iphenyl)-2-aminoethanethiol (Compound 21 (R=4-F), FIG. 5)
(O-1450)
[0153] 2.beta.-(1-Propanoyl)-3.alpha.-(4-fluorophenyl)nortropane 20
(65 mg, 0.22 mmol) was combined with MAMA'-Cl (203 mg, 0.27 mmol,
1.2 eq.), KI (74 mg, 0.45 mmol), and K.sub.2CO.sub.3 (309 mg, 2.2
mmol) in anhydrous MeCN (10 mL) and brought to reflux for 6 h then
cooled to room temperature. The solvent was removed under vacuum
and the residue was partitioned between CHCl.sub.3 and water. The
aqueous layer was extracted with CHCl.sub.3 (3.times.5 mL) and the
combined organic extracts were dried (Na.sub.2SO.sub.4), filtered
and concentrated to yield a yellow oil which was applied to a
chromatography column (15 g SiO.sub.2 eluent 120 mL of 50%
EtOAc/hexanes/5% Et.sub.3N). Fractions containing the product were
combined and concentrated to yield 21 as a light yellow oil (142
mg, 41%).
[0154] R.sub.f 0.7 (5% Et.sub.3N/EtOAc); Elemental analysis:
calculated (0.3H.sub.2O) C, 71.02; H, 6.96; N, 5.26; found C,
71.20; H, 6.45; N, 5.14. .sup.1H-NMR (CDCl.sub.3) .delta. 1.2-3.5
(m, 24H), 2.85 (s, 2H), 3.02 (s, 3H), 3.42 (s, 3H), 6.85-7.0 (m,
2H), 7.1-7.7 (m, 32H).
EXAMPLE 30
N-[(2-((3'-N'-Propyl-(1''R)-3''.alpha.-(4-fluorophenyl)
tropane-2''.beta.-methoxymethylcarbamoyl)
(2-mercaptoethyl)amino)acetyl)-2-aminoethanethiolato]rhenium (V)
oxide (Compound 22 (Re: R=4-F), FIG. 5) (O-1451)
[0155]
N-[2-(3'-N'-propyl-(1''R)-3''.alpha.-(4-fluorophenyl)tropane-2''.b-
eta.-(methoxymethylcarbamoyl))
(2-((triphenylmethyl)thio)ethyl)amino)acetyl]-S-(triphenyl)-2-aminoethane-
thiol 21 (21 mg, 0.02 mmol) in EtOH (4 mL) was heated to reflux. A
solution of SnCl.sub.2 (8.4 mg, 0.04 mmol) in 0.005 M HCl (0.5 mL)
was added quickly followed immediately by NaReO.sub.4 (13 mg, 0.04
mmol) in 0.005 M HCl (0.5 mL). The solution was refluxed for 10 h
and was then loaded onto 0.5 g silica and pumped overnight. The
silica-adsorbed material was applied to a chromatography column (3
g SiO.sub.2, eluent 30% EtOAc/hexanes/5% Et.sub.3N). Like fractions
were combined and concentrated to yield as a purple foam (3.7 mg;
26%).
[0156] R.sub.f 0.5 (25% hexanes in EtOAc); .sup.1H-NMR (CDCl.sub.3)
.delta. 1.1-4.2 (m, 30H), 4.5-4.6 (m, 1H), 4.75, 5.12 (2d, J=16.7
Hz, 2.times.6.beta., 1H), 6.8-7.0 (m, 2H), 7.1-7.2 (m, 2H).
EXAMPLE 31
2.beta.-Methoxymethylcarbamoyl-3.beta.-(4-fluorophenyl)nortropane
(Compound 20A (R=4-F), FIG. 5)
[0157] 2.beta.-(1-Propanoyl)-3.beta.-(4-fluorophenyl)tropane (143
mg, 0.50 mmol) was combined with 1-chloroethyl chloroformate (2 mL)
and the solution was heated to reflux for 2 h. The excess
chloroformate was removed in vacuo and the residue was refluxed in
methanol (30 mL) for 45 min. The methanol was removed in vacuo and
the residue was dissolved in CHCl.sub.3 and
NaHCO.sub.3/Na.sub.2CO.sub.3 (pH=9). The aqueous layer was
extracted CHCl.sub.3 (3.times.10 mL) and the combined organic
extracts were dried (Na.sub.2SO.sub.4), filtered and concentrated.
The residue was chromatographed (10-20% Et.sub.3N in EtOAc). Like
fractions were combined to yield 80 mg (58%).
[0158] .sup.1H-NMR (CDCl.sub.3) .delta. 1.50 (m, 1H), 1.7 (m, 2H),
1.95-2.22 (m, 2H), 2.5 (ddd, 1H), 2.92 (s, 3H), 3.1-3.4 (m, 2H),
3.3 (s, 3H), 3.6 (m, 1H), 3.72 (m, 1H), 6.9-7.0 (m, 2H), 7.1-7.3
(m, 2H).
EXAMPLE 32
N-[2-(3'-N'-Propyl-(1''R)-3''.beta.-(4-fluorophenyl)tropane-2''.beta.-meth-
oxymethylcarbamoyl)((2-((triphenylmethyl)thio)ethyl)amino)acetyl]-S-(triph-
enyl)-2-aminoethanethiol (Compound 21A (R=4-F), FIG. 5)
[0159] 2.beta.-(1-Propanoyl)-3.beta.-(4-fluorophenyl)nortropane (80
mg, 0.28 mmol) was combined with MAMA'-Cl (244 mg, 0.32 mmol,), KI
(84 mg, 0.5 mmol, 2.0 eq.), and K.sub.2CO.sub.3 (322 mg, 2.3 mmol)
in anhydrous MeCN (10 mL) and brought to reflux overnight then
cooled to room temperature. The solvent was removed under vacuum
and the residue was applied to a chromatography column (5%
Et.sub.3N in EtOAc). Fractions containing the product were combined
and concentrated to yield the product as a light yellow foam (80
mg, 29%).
[0160] R.sub.f 0.65 (5% Et.sub.3N in EtOAc); .sup.1H-NMR
(CDCl.sub.3) .delta. 1.2-3.1 (m, 28H), 3.37 (m, 1H), 3.51 (s, 3H),
3.55 (m, 1H), 6.9-7.0 (m, 2H), 7.1-7.5 (m, 32H).
EXAMPLE 33
N-((2-((3'-N'-Propyl-(1''R)-3''.beta.-(4-fluorophenyl)tropane-2''.beta.-me-
thoxymethylcarbamoyl)
(2-mercaptoethyl)amino)acetyl)-2-aminoethanethiolato]rhenium (V)
oxide (Compound 22A (Re: R=4-F), FIG. 5) (O-1451)
[0161]
N-[2-(3'-N'-propyl-(1''R)-3''.beta.-(4-fluorophenyl)tropane-2''.be-
ta.-methoxymethylcarbamoyl)
((2-((triphenylmethyl)thio)ethyl)amino)acetyl]-S-(triphenyl)-2-aminoethan-
ethiol (22 mg, 0.02 mmol) in EtOH (10 mL) was heated to reflux. A
solution of SnCl.sub.2 (8.4 mg, 0.05 mmol) in 0.005 M HCl (1.0 mL)
was added quickly followed immediately by NaReO.sub.4 (13 mg, 0.05
mmol) in 0.005 M HCl (0.5 mL). The solution was refluxed for 10 h
and was then applied to a chromatography column (65%
EtOAc/hexanes/5% Et.sub.3N). Like fractions were combined and
concentrated to yield a foam (10 mg; 65%).
[0162] R.sub.f 0.44 (5% Et.sub.3N, 30% hexanes in EtOAc);
.sup.1H-NMR (CDCl.sub.3) .delta. 1.5-4.2 (m, 30H), 4.4-4.6 (m, 1H),
4.72, 5.05 (2d, J=16.5 Hz, 1H), 6.8-7.0 (m, 2H), 7.1-7.3 (m,
2H).
EXAMPLE 34
(1
R)-N-Methyl-2-hydroxymethyl-3-(4-fluorophenyl)-8-aza-bicyclo[3.2.1]oct--
2-ene (Compound 38 (R=4-F), FIG. 9) (O-1337)
[0163]
(1R)-N-Methyl-2-methoxycarbonyl-3-(4-fluorophenyl)-8-azabicyclo
[3.2.1]oct-2-ene 3 (500 mg, 1.82 mmol) was dissolved in benzene (15
mL) and LAH (70 mg, 1.82 mmol) was added. The reaction was heated
to 60.degree. C. overnight. The reaction was cooled to 0.degree.
C., and water (70 .mu.L), 15% NaOH (70 .mu.L), water (200 .mu.L)
were added. The reaction was stirred for 15 min and then the salts
were filtered off through a pad of celite. The filtrate was dried
over Na.sub.2SO.sub.4 filtered and concentrated to yield 38 (454
mg, 90%) which was purified by column chromatography (30 g
SiO.sub.2; eluent: 4% Et.sub.3N in 10% MeOH/CHCl.sub.3). Like
fractions were combined, reduced and pumped at high vacuum
overnight to yield 38 (336 mg, 74%).
[0164] Mp. 106.6-108.7.degree. C.; R.sub.f 0.2 (10%
MeOH/CHCl.sub.3; 5% Et.sub.3N); .sup.1H-NMR (CDCl.sub.3) .delta.
1.5-1.7 (m, 1H), 1.9-2.0 (m, 3H), 2.1-2.3 (m, 2H), 2.4 (s, 3H),
2.72 (brd, J=18 Hz, 1H), 3.31 (br s, 1H), 3.55 (d, J=4.1 Hz, 1H),
3.89 (d, J=12.3 Hz, 1H), 4.02 (d, J=12.3 Hz, 1H), 6.9-7.0 (m, 2H),
7.1-7.2 (m, 2H). Elemental analysis: calculated C, 72.85; H, 7.34;
N, 5.66; found C, 72.94; H, 7.33; N, 5.73.
EXAMPLE 35
(1R)-N-Methyl-2-hydroxymethyl-3-(3,4-dichlorophenyl)-8-aza-bicyclo[3.2.1]o-
ct-2-ene (Compound 38 (R=3,4-Cl.sub.2), FIG. 9)
[0165]
(1R)-N-Methyl-2-methoxycarbonyl-3-(3,4-dichlorophenyl)-8-azabicycl-
o[3.2.1]-oct-2-ene 3 (508 mg, 1.56 mmol) was dissolved in benzene
(20 mL) and LAH (62 mg, 1.56 mmol) was added. The reaction was
heated to reflux overnight. The reaction was cooled to 0.degree.
C., and water (80 .mu.L), 10% KOH (90 .mu.L), water (300 .mu.L)
were added.
[0166] The reaction was stirred for 15 min and then the salts were
filtered off through a pad of celite. The filtrate was dried over
Na.sub.2SO.sub.4 filtered and concentrated to yield a yellow foam
(420 mg, 90%) and purified by column chromatography (3%
Et.sub.3N/7% MeOH in CHCl.sub.3) to give 280 mg (60%).
[0167] R.sub.f 0.2 (5% MeOH/CHCl.sub.3; 3% Et.sub.3N); .sup.1H-NMR
(CDCl.sub.3) .delta. 1.5-1.66 (m, 1H), 1.84-1.98 (m, 2H), 2.0 (m,
2H), 2.08-2.24 (m, 2H), 2.39 (s, 3H), 2.7 (dd, 1H), 3.3 (m, 1H),
3.54 (d, J=5.5 Hz, 1H), 3.89 (d, J=11.8 Hz, 1H), 4.01 (d, J=11.8
Hz, 1H), 6.99 (dd, 1H), 7.25 (d, 1H), 7.37 (d, 1H).
EXAMPLE 36
(1R)-N-Methyl-2-carbonyl-3-(4-fluorophenyl)-8-azabicyclo[3.2.1]-oct-2-ene
(Compound 39 (R=4-F), FIG. 9)
[0168] A solution of (COCl).sub.2 (55 mg, 40 .mu.L, 0.43 mmol) and
CH.sub.2Cl.sub.2 (2 mL) was cooled to -78.degree. C. and DMSO (65
.mu.L) was added dropwise over 3 min. The reaction was stirred for
a further 5 min at -78.degree. C. and compound 38 (90 mg, 0.36
mmol) in CH.sub.2Cl.sub.2 (2 mL) was added dropwise over 10 min.
After a further 20 min, Et.sub.3N (250 .mu.L, 1.80 mmol) was added
over 30 min. The reaction was stirred for a further 10 min and then
allowed to warm to room temperature. CH.sub.2Cl.sub.2 (20 mL) and
1M NaOH was added, the layers partitioned and the aqueous layer was
washed with CH.sub.2Cl.sub.2 (1.times.15 mL). Combined organic
extracts were dried over Na.sub.2SO.sub.4, filtered, concentrated
and pumped at high vacuum. The yellow oil obtained (86 mg) was
purified by column chromatography (4.5g SiO.sub.2; 30%
EtOAc/hexanes/5% Et.sub.3N) and like fractions were combined,
concentrated and dried at high vacuum to yield the product 39 (62
mg, 63%).
[0169] R.sub.f 0.2 (20% EtOAc/hexane, 5% Et.sub.3N); .sup.1H-NMR
(CDCl.sub.3) .delta. 1.55-1.65 (m, 1H), 1.72-1.82 (m, 2H), 2.14-2.3
(m, 2H), 2.40 (s, 3H), 2.9-3.0 (m, 1H), 3.36-3.44 (m, 1H),
4.02-4.30 (m, 1H),7.0-7.1 (m, 2H), 7.16-7.24 (m, 2H), 9.45 (s,
1H).
EXAMPLE 37
(1R)-N-Methyl-2-carbonyl-3-(3,4-dichlorophenyl)-8-azabicyclo[3.2.1]oct-2-e-
ne (Compound 39 (R=3,4-Cl.sub.2), FIG. 9)
[0170] A solution of (COCl).sub.2 (125 mg, 87 .mu.L, 1.0 mmol) and
CH.sub.2Cl.sub.2 (20 mL) was cooled to -78.degree. C. and DMSO (150
.mu.L) was added dropwise over 3 min. The reaction was stirred for
a further 5 min at -78.degree. C. and compound 38 (245 mg, 0.82
mmol) in CH.sub.2Cl.sub.2 (5 mL) was added dropwise over 7 min.
After a further 30 min, Et.sub.3N (600 .mu.L, 4.0 mmol) was added
over about 15 min. The reaction was stirred for a further 10 min.
and then allowed to warm to room temperature. CH.sub.2Cl.sub.2 (20
mL) and Na.sub.2CO.sub.3 (20 mL; 1M) was added, the layers
partitioned and the aqueous layer was washed with CH.sub.2Cl.sub.2
(1.times.20 mL). Combined organic extracts were dried
Na.sub.2SO.sub.4, filtered, concentrated and pumped at high vacuum.
The residue was purified by column chromatography (25g SiO.sub.2;
60% EtOAc/hexanes/5% Et.sub.3N) and like fractions were combined,
concentrated and dried at high vacuum to yield the product 39 (154
mg, 63%).
[0171] R.sub.f 0.2 (75% EtOAc/hexane, 5% Et.sub.3N); .sup.1H-NMR
(CDCl.sub.3) .delta. 1.5-1.7 (m, 1H), 1.7-1.85 (m, 1H), 2.1-2.3 (m,
2H), 2.38 (s, 3H), 2.93 (dd, 1H), 3.39 (m, 1H), 4.05 (dd, 1H),7.08
(dd, 1H), 7.34 (d, 1H), 7.46 (d, 1H), 9.45 (s, 1H).
EXAMPLE 38
(1R)-N-Methyl-2-(2-hyroxypropyl)-3-(4-fluorophenyl)-8-azabicyclo[3.2.1]oct-
-2-ene (Compound 40 (R=4-F), FIG. 9)
[0172] Compound 39 (62 mg, 0.26 mmol) was dissolved in anhydrous
THF at room temperature and EtMgBr/Et.sub.2O (3M; 900 .mu.l) was
added dropwise. The reaction was stirred at room temperature for 20
min and then at 65.degree. C. overnight, and then added slowly to a
mixture of 1M HCl (50 mL) and Et.sub.2O (50 mL) at 0.degree. C. The
mixture was basified with saturated aq. Na.sub.2CO.sub.3 and
separated. The aqueous layer was extracted with CHCl.sub.3
(2.times.20 mL), dried (Na.sub.2SO.sub.4), filtered, evaporated and
dried at high vacuum. The residue (90 mg) was purified by column
chromatography (6 g SiO.sub.2; 5% Et.sub.3N/EtOAc) and like
fractions were combined, evaporated and dried at high vacuum to
yield 40 (82 mg, 52%).
[0173] R.sub.f 0.4 (10% MeOH/CHCl.sub.3/5% Et.sub.3N); .sup.1H-NMR
(CDCl.sub.3) .delta. 0.83 (t, J=7.6 Hz, 3H), 1.4-1.7 (m, 4H), 1.81
(d, J=18 Hz, 1H), 2.0 (m, 1H), 2.1-2.3 (m, 2H), 2.42 (s, 3H), 2.7
(m, 1H), 3.3 (m, 1H), 3.53 (m, 1H), 4.1-4.2 (m, 1H), 6.9-7.1 (m,
4H)
EXAMPLE 39
(1R)-N-Methyl-2-(2-hyroxypropyl)-3-(3,4-dichlorophenyl)-8-azabicyclo[3.2.1-
]oct-2-ene (Compound 40 (R=3,4-Cl.sub.2), FIG. 9)
[0174] Compound 39 (150 mg, 0.5 mmol) was dissolved in anhydrous
THF (25 mL) at room temperature and EtMgBr/Et.sub.2O (3M; 350
.mu.l, 1.05 mmol) was added dropwise over 2 min. The reaction was
stirred at room temperature for 2.75 h and then added slowly to a
mixture of 1M HCl (20 mL) and Et.sub.2O (20 mL). The mixture was
basified with saturated aq. Na.sub.2CO.sub.3 and separated. The
aqueous layer was extracted with CHCl.sub.3 (3.times.20 mL) dried
(Na.sub.2SO.sub.4), filtered, evaporated and dried at high vacuum.
The residue (197 mg) was purified by column chromatography (20 g
SiO.sub.2; 5% Et.sub.3N/EtOAc) and like fractions were combined,
evaporated and dried at high vacuum to yield 40 (109 mg, 66%).
[0175] R.sub.f 0.07 (5% Et.sub.3N, 20% hexane in EtOAc);
.sup.1H-NMR (CDCl.sub.3) .delta. 0.85 (t, J=7.4 Hz, 3H), 1.4-1.7
(m, 4H), 1.79 (d, J=17.6 Hz, 1H), 1.95-2.1 (m, 1H), 2.1-2.3 (m,
2H), 2.42 (s, 3H), 2.68 (dd, J=4, 18 Hz, 1H), 3.3 (m, 1H), 3.38 (d,
1H), 3.52 (d, J=5 Hz, 1H), 4.1-4.2 (m, 1H), 6.94 (dd, 1H), 7.20
(dd, 1H).
EXAMPLE 40
(1R)-2-Propanoyl-3-(4-fluorophenyl)-8-azabicyclo[3.2.1]-oct-2-ene
(Compound 26 (R=4-F), FIG. 9)
[0176] Tropane 40 (28 mg, 0.1 mmol) in CH.sub.2Cl.sub.2 was added
dropwise to a cold (-78.degree. C.) solution of (COCl).sub.2 (5.5
.mu.L, 0.12 mmol) and DMSO (20 .mu.L, 0.26 mmol) in
CH.sub.2Cl.sub.2. After stirring for 30 min at -78.degree. C.,
Et.sub.3N (75 .mu.L, 0.5 mmol) was added and the reaction was
warmed to room temperature. CH.sub.2Cl.sub.2 and 1M NaOH were added
and the layers separated. The organic layer was dried
(Na.sub.2SO.sub.4), filtered and evaporated and the product applied
to a chromatography column (5% Et.sub.3N, 25% hexane, 70% EtOAc) to
give 18.6 mg of product (66%).
[0177] R.sub.f 0.2 (5% Et.sub.3N, 25% Hexanes, 70% EtOAc);
.sup.1H-NMR (CDCl.sub.3) .delta. 0.80 (t, 3H), 1.6 (m, 1H), 1.8-2.3
(m, 6H), 2.40 (s, 3H), 2.75 (dd, 1H), 3.35 (m, 1H), 3.73 (m, 1H),
7.0 (m, 2H), 7.05-7.15 (m, 2H).
EXAMPLE 41
(1R)-2-Methoxycarbonyl-3-(4-fluorophenyl)-8-norazabicyclo[3.2.1]-oct-2-ene
(Compound 29 (R=4-F), FIG. 7) (O-1131)
[0178] 2-Methoxycarbonyl-3-(4-fluorophenyl)tropene 3 (362 mg, 1.3
mmol) was combined with 1-chloroethyl chloroformate (1 mL) and the
solution was heated to reflux for 2 h. The excess chloroformate was
removed in vacuo and the residue was refluxed in methanol (30 mL)
for 45 min. The methanol was removed in vacuo and the residue was
dissolved in CHCl and NaHCO.sub.3/Na.sub.2CO.sub.3 (pH=9). The
aqueous layer was extracted CHCl (3.times.10 mL) and the combined
organic extracts were dried (Na.sub.2SO.sub.4), filtered and
concentrated. The residue was chromatographed (eluent: 1-3%
Et.sub.3N in EtOAc+0.5% NH.sub.4OH). Like fractions were combined
to yield a yellow solid (234 mg, 68%).
[0179] R.sub.f 0.7 (10% MeOH in hexanes+0.5% NH.sub.4OH); mp.
67-68.degree. C.; .sup.1H-NMR (CDCl.sub.3) .delta. 1.50-3.0 (m,
6H), 3.8 (m, 1H), 4.2 (m, 1H), 6.8-7.2 (m, 4H). Elemental analysis:
calculated C, 68.95; H, 6.17; N, 5.36; found C, 68.81; H, 6.24; N,
5.40.
EXAMPLE 42
(1R)-2-Methoxycarbonyl-3-(3,4-dichlorophenyl)-8-norazabicyclo[3.2.1]oct-2--
ene (Compound 29 (R=3,4-Cl.sub.2), FIG. 7) (O-1130)
[0180] 2-Methoxycarbonyl-3-(3,4-dichlorophenyl)tropene (200 mg,
0.61 mmol) was combined with 1-chloroethyl chloroformate (4 mL) and
the solution was heated to reflux for 2 h. The excess chloroformate
was removed in vacuo and the residue was refluxed in methanol (30
mL) for 45 min. The methanol was removed in vacuo and the residue
was dissolved in CHCl.sub.3 and NaHCO.sub.3/Na.sub.2CO.sub.3
(pH=9). The aqueous layer was extracted CHCl.sub.3 (3.times.10 mL)
and the combined organic extracts were dried (Na.sub.2SO.sub.4),
filtered and concentrated. The residue was chromatographed (eluent:
5-15% Et.sub.3N in EtOAc). Like fractions were combined to yield a
yellow oil.
[0181] R.sub.f 0.3 (10% Et.sub.3N in EtOAc); .sup.1H-NMR
(CDCl.sub.3) .delta. 1.50-2.3 (m, 5H), 2.5-2.9 (m, 1H), 3.53 (s,
3H), 3.8 (m, 1H), 4.2 (m, 1H), 6.9 (dd, 1H), 7.2 (dd, 1H), 7.4 (d,
1H). Elemental analysis: calculated C, 57.71; H, 4.84; N, 4.49;
found C, 57.45; H, 4.87; N, 4.47.
EXAMPLE 43
N-[2-(3'-N'-Propyl-(1''R)-3''-(4-fluorophenyl)trop-2-ene-2''-(methoxycarbo-
nyl))((2-((triphenylmethyl)thio)ethyl)amino)acetyl]-S-(triphenyl)-2-aminoe-
thanethiol (Compound 30 (R=4-F), FIG. 7)
[0182] 2-(1-Methoxycarbonyl)-3-(4-fluorophenyl)nortrop-2-ene (134
mg, 0.5 mmol) was combined with MAMA'-Cl (387 mg, 0.5 mmol,), KI
(85 mg, 0.5 mmol), and K.sub.2CO.sub.3 (707 mg, 5 mmol) in
anhydrous MeCN (10 mL) and brought to reflux overnight then cooled
to room temperature. The solvent was removed under vacuum and the
residue was applied to a chromatography column (1-6% Et.sub.3N in
EtOAc). Fractions containing the product were combined and
concentrated to yield the product as a light yellow foam (148 mg,
29%).
[0183] R.sub.f 0.18 (10% Et.sub.3N in hexane); .sup.1H-NMR
(CDCl.sub.3) .delta. 1.3-3.15 (m, 22H), 3.2-3.4 (m, 1H), 3.5 (s,
3H), 3.8-3.9 (m, 1H), 6.8-7.8 (m, 34H).
EXAMPLE 44
N-[2-(3'-N'-Propyl-(1''R)-3''-(3,4-dichlorophenyl)trop-2-ene-2''-(methoxyc-
arbonyl)((2-((triphenylmethyl)
thio)ethyl)amino)acetyl]-S-(triphenyl)-2-aminoethanethiol (Compound
30 (R=3,4-Cl.sub.2), FIG. 7)
[0184] 2-(1-Methoxycarbonyl)-3-(3,4-dichlorophenyl)nortrop-2-ene
(107 mg, 0.34 mmol) was combined with MAMA'-Cl (258 mg, 0.34
mmol,), KI (57 mg, 0.34 mmol), and K.sub.2CO.sub.3 (472 mg, 3.4
mmol) in anhydrous MeCN (10 mL) and brought to reflux overnight
then cooled to room temperature. The solvent was removed under
vacuum and the residue was applied to a chromatography column (1-6%
Et.sub.3N in EtOAc). Fractions containing the product were combined
and concentrated to yield the product as a foam (111 mg, 31%).
[0185] R.sub.f 0.18 (5% Et.sub.3N in hexane); .sup.1H-NMR
(CDCl.sub.3) .delta. 1.4-3.2 (m, 22H), 3.2-3.4 (m, 1H), 3.5 (s,
3H), 3.8-3.9 (m, 1H), 6.8-7.6 (m, 33H).
EXAMPLE 45
N-[(2-((3'-N'-Propyl-(1''R)-3''-(4-fluorophenyl)trop-2-ene-2''-methoxycarb-
onyl) (2-mercaptoethyl)amino)acetyl)-2-aminoethanethiolato]rhenium
(V) oxide (Compound 31 (Re: R=4-F), FIG. 7) (O-1135)
[0186] A solution of
N-[2-(3'-N'-propyl-(1''R)-3''-(4-fluorophenyl)trop-2-ene-2''-(methoxycarb-
onyl) (2-((triphenylmethyl)
thio)ethyl)amino)acetyl]-S-(triphenyl)-2-aminoethanethiol (130 mg,
0.13 mmol) in EtOH (10 mL) was heated to reflux. A solution of
SnCl.sub.2 (28 mg, 0.15 mmol) in 0.005 M HCl (1.0 mL) was added
quickly followed immediately by NaReO.sub.4 (40 mg, 0.15 mmol) in
0.005 M HCl (0.5 mL). The solution was refluxed for 10 h and was
then applied to a chromatography column (1-10% Et.sub.3N in EtOAc).
Like fractions were combined and concentrated to yield a foam (34
mg; 37%).
[0187] R.sub.f 0.09 (10% Et.sub.3N in EtOAc); .sup.1H-NMR
(CDCl.sub.3) .delta. 1.4-4.3 (m, 23H), 3.5 (s, 3H), 4.5-4.9 (m,
2H), 6.9-7.2 (m, 4H). Elemental analysis: calculated C, 41.49; H,
4.50; N, 6.05; found C, 41.77; H, 4.44; N, 5.93. Accurate mass
calculated 696.1348, found 696.1405.
EXAMPLE 46
N-[(2-((3'-N'-Propyl-(1''R)-3''-(3,4-dichlorophenyl)trop-2-ene-2''-methoxy-
carbonyl)
(2-mercaptoethyl)amino)acetyl)-2-aminoethanethiolato]rhenium (V)
oxide (Compound 31 (Re: R=3,4-Cl.sub.2), FIG. 7) (O-1136)
[0188] A solution of
N-[2-(3'-N'-propyl-(1''R)-3''-(3,4-dichlorophenyl)trop-2-ene-2''-(methoxy-
carbonyl) (2-((triphenylmethyl)
thio)ethyl)amino)acetyl]-S-(triphenyl)-2-aminoethanethiol (100 mg,
0.1 mmol) in EtOH (10 mL) was heated to reflux. A solution of
SnCl.sub.2 (20 mg, 0.1 mmol) in 0.005 M HCl (1.0 mL) was added
quickly followed immediately by NaReO.sub.4 (29 mg, 0.11 mmol) in
0.005 M HCl. (0.5 mL). The solution was refluxed for 10 h and was
then applied to a chromatography column (1-10% Et.sub.3N in EtOAc).
Like fractions were combined and concentrated to yield a foam (56
mg; 78%).
[0189] R.sub.f 0.09 (10% Et.sub.3N in EtOAc); .sup.1H-NMR
(CDCl.sub.3) .delta. 1.4-4.2 (m, 23H), 3.55 (s, 3H), 4.4-4.9 (m,
2H), 6.95 (dd, 1H), 7.2 (d, 1H), 7.4 (d, 1H). Elemental analysis:
calculated C, 38.65; H, 4.05; N, 5.63; found C, 38.91; H, 3.96; N,
5.57. Accurate mass calculated 746.0663, found 746.0689.
EXAMPLE 47
N-[2-(3'-N'-Propyl-(1''R-3''.beta.-(4-fluorophenyl)-2''.beta.-methoxycarbo-
nyltropane))
((2-((triphenylmethyl)thio)ethyl)amino)acetyl]-2-aminoethanethiol
(Compound 33 (R=4-F), FIG. 8)
[0190] To a solution of
nor-3.beta.-(4-fluorophenyl)-2.beta.-methoxycarbonyltropane (52.6
mg, 0.2 mmol) in dry acetonitrile (10 mL) was added in succession
N-[2-((3-chloropropyl)-(2-((triphenylmethyl)thio)ethyl)amino)acetyl]-S-(t-
riphenylmethyl)-2-aminoethanethiol (151 mg, 0.2 mmol), KI (33 mg,
0.2 mmol), and K.sub.2CO.sub.3 (280 mg, 2.0 mmol). The resulting
slurry was then boiled overnight. Once the reaction was complete
then the solution was allowed to cool to room temperature and then
2 g of silica gel was added and the solvent evaporated. The
resulting solid was layered onto a silica gel column and eluted
with 0.5% NH.sub.4OH in 1:1 solution of EtOAc and hexanes. The
title compound was recovered as a foam in 72% yield (141 mg). This
was converted to the dihydrochloride.
[0191] Mp. 166-168.degree. C.; IR (KBr) 1666 cm.sup.-1; .sup.1H-NMR
(CDCl.sub.3) .delta. 1.8-3.8, (m, 24H), 3.3 (s, 3H), 3.9-4.0 (m,
1H), 4.2-4.3 (m, 1H), 4.4-4.5 (m, 1H), 6.9-7.4 (m, 34 H). Elemental
analysis: calculated (2 HCl.2 H.sub.2O): C, 68.24; H, 6.47; N,
3.85. Found: C, 38.03; H, 6.40; N, 3.82.
EXAMPLE 48
(RS)-N-[2-((3'-N'-Propyl-(1''R-3''.beta.-(3,4-dichlorophenyl)-2''.beta.-me-
thoxycarbonyltropane))
(2-mercaptoethyl)amino)acetyl)-2-aminoethanethiol (Compound 33
(R=3,4-Cl.sub.2), FIG. 8) (O-863)
[0192] Prepared identically to the compound in Example 47
above.
[0193] Mp. 108.degree. C., IR (KBr) 1724, 1653, 957 cm.sup.-1;
.sup.1H-NMR (CDCl.sub.3) .delta. 1.5-3.9 (m, 22H), 3.55 & 3.50
(2s, 3H), 3.9-4.2 (m, 2H), 4.5-4.7 (m, 1H), 4.80 (d, 16.4 Hz, 1H),
7.0-7.4 (m, 3H); Accurate mass calculated for
C.sub.24H.sub.33Cl.sub.2N.sub.3S.sub.2O.sub.4Re [MH].sup.+748.0819,
found 748.0856. Elemental analysis: calculated, C, 38.55; H, 4.31;
N, 5.62. Found C, 38.79; H, 4.38; N, 5.41.
EXAMPLE 49
(RS)-N-[2-((3'-N'-Propyl-(1''R-3''.beta.-(4-fluorophenyl)-2''.beta.-methox-
ycarbonyltropane))
(2-mercaptoethyl)amino)acetyl)-2-aminoethanethiolato]rhenium (V)
oxide (Compound 34 (Re: R=4-F), FIG. 8) (O-861)
[0194]
N-[2-((3'-N'-Propyl-3''.beta.-(4-fluorophenyl)tropane-2''.beta.-ca-
rboxylic acid methyl ester)
(2-((triphenylmethyl)thio)ethyl)amino)acetyl]-S-(triphenylmethyl)-2-amino-
ethanethiol (98 mg, 0.1 mmol) were dissolved in boiling ethanol
(1.5 mL). To this was added a solution of SnCl.sub.2 (21 mg, in 200
mL of 0.05 M HCl), followed immediately by a solution of
NaReO.sub.4 (30 mg in 200 mL of 0.05 M HCl). Boiling was continued
overnight, after which boiling CH.sub.3CN (10 mL) was added and the
resulting solution filtered through a pad of celite. The cake was
further washed two more times with boiling CH.sub.3CN (2.times.20
mL). To the filtrate was added silica gel (1 g) and the solvent
evaporated. The solid was then layered onto a silica gel column and
eluted with EtOAc. The title compound was isolated as a mixture of
diastereomers in 90% yield (608 mg).
[0195] Mp 101.9.degree. C., IR (KBr) 1720, 1666, 957 cm.sup.-1;
.sup.1H-NMR (CDCl.sub.3) .delta. 1.4-4.2, (m, 24H), 3.46 & 3.5
(2s, 3H), 4.4-4.7 (m, 1H), 4.80 & 4.82 (2d, 16 Hz, 1H), 6.8-7.3
(m, 4H); Accurate mass calculated for
C.sub.24H.sub.34FN.sub.3S.sub.2O.sub.4Re [MH].sup.+698.1505, found
698.1557. This was converted to a hydrochloride for analysis:
Elemental analysis: calculated (HCl.2 H.sub.2O) C, 37.47; H, 4.98;
N, 5.46. Found C, 37.45; H, 4.95; N, 5.40.
EXAMPLE 50
(RS)-N-[2-((3'-N'-Propyl-(1''R-3''.beta.-(3,4-dichlorophenyl)-2''.beta.-me-
thoxycarbonyltropane))
(2-mercaptoethyl)amino)acetyl)-2-aminoethanethiolato]rhenium (V)
oxide (Compound 34 (Re: R=3,4-Cl.sub.2), FIG. 8) (O-863)
[0196] Prepared identically to the compound in Example 49
above.
[0197] Mp. 108.degree. C., IR (KBr) 1724, 1653, 957 cm.sup.-1;
.sup.1H-NMR (CDCl.sub.3) .delta. 1.5-3.9 (m, 22H), 3.55 & 3.50
(2s, 3H), 3.9-4.2 (m, 2H), 4.5-4.7 (m, 1H), 4.80 (d, 16.4 Hz, 1H),
7.0-7.4 (m, 3H); Accurate mass calculated for
C.sub.24H.sub.33Cl.sub.2N.sub.3S.sub.2O.sub.4Re [MH].sup.+748.0819,
found 748.0856. Elemental analysis: calculated, C, 38.55; H, 4.31;
N, 5.62. Found C, 38.79; H, 4.38; N, 5.41.
EXAMPLE 51
(1R)-2.beta.-Methoxycarbonyl-3.alpha.-(3,4-dichlorophenyl)-8-azabicyclo[3.-
2.1]octane (Compound 35 (R=3,4-Cl.sub.2), FIG. 8)
[0198]
(1R)-N-Methyl-2.beta.-methoxycarbonyl-3.alpha.-(3,4-dichlorophenyl-
)-8-azabicyclo[3.2.1]-octane 12 (375 mg, 1.14 mmol) and
.alpha.-chloroethyl chloroformate (ACE-Cl) (3 mL) were combined and
heated at 100.degree. C. (oil bath temperature) for 1 h. Excess
ACE-Cl was then removed under reduced pressure, and methanol (50
mL) was added to the residue. The mixture was then heated at reflux
for 30 min and then concentrated to dryness. The residue obtained
was dissolved in CH.sub.2Cl.sub.2 (75 mL), washed with aqueous
NH.sub.4OH, dried over Na.sub.2SO.sub.4, filtered, and concentrated
to afford the crude demethylated product. Purification by flash
chromatography (1% NH.sub.4OH, 50-0% hexanes, 50-90% EtOAc 0-10%
methanol) gave 250 mg (70%) of 35.
[0199] R.sub.f 0.14, (5% Et.sub.3N/EtOAc/hexanes 1:1); .sup.1H-NMR
(CDCl.sub.3) .delta. 1.1-2.5 (m, 8H), 2.9-3.3 (m, 1H), 3.5-3.8 (m,
1H), 3.6 (s, 3H), 6.95-7.40 (m, 3H).
EXAMPLE 52
(1
R)-2.beta.-Methoxycarbonyl-3.alpha.-(4-fluorophenyl)-8-azabicyclo[3.2.1-
]-octane (Compound 35 (R=4-F), FIG. 8)
[0200]
(1R)-N-Methyl-2-methoxycarbonyl-3.alpha.-(4-fluorophenyl)-8-azabic-
yclo[3.2.1]-octane (95 mg, 0.34 mmol) and ACE-Cl (7 mL) were
combined and heated at 100.degree. C. (oil bath temperature) for 1
h. Excess ACE-Cl was then removed under reduced pressure, and
methanol (50 mL) was added to the residue. The mixture was then
heated at reflux for 30 min and then concentrated to dryness. The
residue obtained was dissolved in CH.sub.2Cl.sub.2 (75 mL), washed
with aqueous NH.sub.4OH, dried over Na.sub.2SO.sub.4, filtered, and
concentrated to afford the crude demethylated product. Purification
by flash chromatography (0-5% NH.sub.4OH, 10% MeOH in EtOAc) gave
86 mg (95%) of 35.
[0201] R.sub.f 0.66, (10% MeOH/EtOAc+0.5% NH.sub.4OH); .sup.1H-NMR
(CDCl.sub.3) .delta. 1.2 (ddd, 1H), 1.2-2.8 (m, 5H), 3.3-3.6 (m,
2H), 3.5 (s, 3H), 3.8-4.2 (m, 2H), 6.9-7.3 (m, 4H).
EXAMPLE 53
N-[2-(3'-N'-Propyl-(1''R)-3''.beta.-(3,4-dichlorophenyl)-2''.beta.-methoxy-
carbonyltropane))
((2-((triphenylmethyl)thio)ethyl)amino)acetyl]-S-(triphenyl)-2-aminoethan-
ethiol (Compound 36 (R=3,4-Cl.sub.2), FIG. 8)
[0202] To a solution of
(1R)-2.beta.-methoxycarbonyl-3.alpha.-(3,4-dichlorophenyl)-8-azabicyclo
[3.2.1]octane 35 (250 mg, 0.79 mmol) in dry CH.sub.3CN (40 mL) was
added in succession N-[[[2-[2-(triphenylmethyl)
thio]ethyl](N'-3'-chloropropyl)amino]acetyl]-S-(triphenylmethyl)-2-aminoe-
thanethiol (601 mg, 0.79 mmol), KI (132 mg), and K.sub.2CO.sub.3
(1.1 g). The resulting slurry was maintained at reflux overnight.
Once the reaction was complete, the solution was allowed to cool to
room temperature and then partitioned between concentrated aqueous
NH.sub.4OH and CH.sub.2Cl.sub.2. The layers were separated and the
organic phase dried with Na.sub.2SO.sub.4. The solution was
filtered and concentrated and the residue purified by flash
chromatography (15 g, SiO.sub.2; 0-5% Et.sub.3N in a 1:1 mixture of
EtOAc/hexanes) which gave 100 mg (12%) of a white foam.
[0203] R.sub.f 0.26 (2.5% NH.sub.4OH in EtOAc/hexanes 1/1);
.sup.1H-NMR (CDCl.sub.3) .delta. 1.20-1.54 (m, 5H), 1.80-2.50 (m,
15H), 2.81 (d, J=17 Hz, 1H), 2.88 (d, J=17 Hz, 1H), 3.00 (m, 2H),
3.16-3.40 (m, 3H), 3.54 (s, 3H), 7.03 (dd, J=2.2, 8.5 Hz, 1H),
7.14-7.53 (m, 32H). Elemental analysis: calculated, C, 72.07; H,
6.15; N, 4.07; Cl, 6.86; found C, 72.18; H, 6.21; N, 3.97; Cl,
6.75.
EXAMPLE 54
N-[2-(3'-N'-Propyl-(1''R)-3''.alpha.-(4-fluorophenyl)-2''.beta.-methoxycar-
bonyl-tropane)) ((2-((triphenylmethyl)
thio)ethyl)amino)acetyl]-S-(triphenyl)-2-aminoethanethiol (Compound
36 (R=4-F), FIG. 8)
[0204] To a solution of
(1R)-2.beta.-methoxycarbonyl-3.alpha.-(4-fluorophenyl)-8-azabicyclo
[3.2.1]octane 35 (230 mg, 0.87 mmol) in dry CH.sub.3CN (25 mL) was
added in succession N-[[[2-[2-(triphenylmethyl)
thio]ethyl](N'-3'-chloropropyl)amino]acetyl]-S-(triphenylmethyl)-2-aminoe-
thanethiol (660 mg, 0.87 mmol), KI (145 mg, 0.87 mmol), and
K.sub.2CO.sub.3 (1.21 g, 8.7 mmol). The resulting slurry was then
maintained at reflux overnight. Once the reaction was complete, the
solution was allowed to cool to room temperature and then
partitioned between concentrated aqueous NH.sub.4OH and
CH.sub.2Cl.sub.2. The layers were separated and the organic phase
dried with Na.sub.2SO.sub.4. The solution was filtered and
concentrated and the residue purified by flash chromatography (50%
EtOAc/hexanes+1% NH.sub.4OH) which gave 435 mg (51%) of a foam.
[0205] R.sub.f 0.45 (1% Et.sub.3N, 50% EtOAc, 49% hexanes);
.sup.1H-NMR (CDCl.sub.3) .delta. 1.20 1.3 (m, 1H), 1.3-2.5 (m,
19H), 2.81 (d, J=17 Hz, 1H), 2.86 (d, J=17 Hz, 1H), 2.95-3.05 (m,
2H), 3.10-3.4 (m, 3H), 3.50 (s, 3H), 6.89-6.96 (m, 2H), 7.1-7.5 (m,
32H). Elemental analysis: calculated (0.75 H.sub.2O), C, 74.78; H,
6.63; N, 4.22; found C, 74.80; H, 6.64; N, 4.17.
EXAMPLE 55
N-[(2-((3'-N'-Propyl-(1''R)-3''.alpha.-(4-fluorophenyl)-2''.beta.-methoxyc-
arbonyltropane)
(2-mercaptoethyl)amino)acetyl)-2-aminoethanethiolato]-rhenium (V)
oxide (Compound 37 (R=4-F), FIG. 8) (O-1186)
[0206] A solution of
N-[2-(3'-N'-propyl-(1''R)-3''.alpha.-(4-fluorophenyl)-2''.beta.-methoxyca-
rbonyltropane) (2-((triphenylmethyl)
thio)ethyl)amino)acetyl]-S-(triphenyl)-2-aminoethane-thiol (226 mg,
0.23 mmol) in EtOH (10 mL) was heated to reflux. A solution of
SnCl.sub.2 (48 mg, 0.25 mmol) in 0.05 M HCl (1.0 mL) was added
quickly followed immediately by NaReO.sub.4 (69 mg, 0.25 mmol) in
0.05 M HCl (0.5 mL). The solution was refluxed for 10 h and was
then applied to a chromatography column (5% Et.sub.3N in
Et.sub.2O). Like fractions were combined and concentrated to yield
a foam (33 mg; 21%).
[0207] R.sub.f 0.38 (10% Et.sub.3N in EtOAc); .sup.1H-NMR
(CDCl.sub.3) .delta. 0.9-4.3 (m, 24H), 3.53, 3.56 (2s, 3H), 4.4-4.7
(m, 1H), 4.75, 5.00 (2s, 1H), 6.8-7.4 (m, 4H). Elemental analysis:
calculated (2/7 Et.sub.2O) C, 42.06; H, 5.03; N, 5.85; found C,
42.08; H, 4.93; N, 5.85.
EXAMPLE 56
N-[(2-((3'-N'-Propyl-(1''R)-3''.alpha.-(3,4-dichlorophenyl)-2''.beta.-meth-
oxycarbonyltropane)
(2-mercaptoethyl)amino)acetyl)-2-aminoethanethiolato]rhenium (V)
oxide (compound 37 (R=3,4-Cl.sub.2), FIG. 8) (O-1196)
[0208] Prepare identically to the compound in Example 55.
[0209] R.sub.f 0.51 (10% Et.sub.3N/EtOAc); .sup.1H-NMR (CDCl.sub.3)
.delta. 1.2-1.4 (m, 1H), 1.4-2.5 (m, 11H), 2.89 (m, 1H), 3.10-3.40
(m, 5H), 3.56 & 3.60 (2s, 3H), 3.6-3.8 (m, 0.5H), 3.90-4.15 (m,
3.5H), 4.60 (m, 1H), 4.73 & 4.95 (2d, J=16.2 & 16.2 Hz,
2.times.0.5H), 7.0-7.1 (m, 1H), 7.24-7.36 (m, 2H). Elemental
analysis: calculated (H.sub.2O), C, 37.64; H, 4.48; N, 5.49; found
C, 37.59; H, 4.31; N, 5.55.
EXAMPLE 57
(1R)-N-Methyl-2-methoxycarbonyl-3-(2-naphthyl)-8-azabicyclo[3.2.1]oct-2-en-
e (Compound 50, FIG. 10)
[0210]
(1R)-2-(Methoxycarbonyl)-3-[[(trifluoromethyl)sulfonyl]oxy]trop-2--
ene 2 (500 mg, 1.52 mmol), LiCl (142 mg, 3.34 mmol),
Pd.sub.2dba.sub.3 (56 mg, 0.06 mmol), Na.sub.2CO.sub.3 (2.0 M
solution in water, 2 mL), diethoxymethane (6 mL) were all charged
to a flask and stirred vigorously. To this solution was added
2-naphthyl boronic acid (340 mg, 1.97 mmol). The reaction was then
brought to reflux for two hours and then filtered through celite.
The cake was washed with ether and all the organic solution was
washed with concentrated ammonium hydroxide solution. The washed
solvent was dried with potassium carbonate, filtered, and
evaporated. The residue was charged to a column (1-4%
Et.sub.3N/EtOAc) and gave 240 mg (51%) of compound 50.
[0211] R.sub.f 0.48 (10% Et.sub.3N/EtOAc). .sup.1H-NMR (CDCl.sub.3)
.delta. 1.3-3.6 (m, 7H), 2.5 (s, 3H), 3.45 (s, 3H), 3.8-4.0 (m,
1H), 7.2-8.0 (m, 7H). Elemental analysis: calculated (0.5 H.sub.2O)
C, 77.24; H, 6.94; N, 4.50; found C, 77.27; H, 6.94; N, 4.48.
EXAMPLE 58
(1R)-2-Methoxycarbonyl-3-(2-naphthyl)-8-azabicyclo[3.2.1]-oct-2-ene
(Compound 51, FIG. 10)
[0212]
(1R)-N-Methyl-2-methoxycarbonyl-3-(2-naphthyl)-8-azabicyclo[3.2.1]-
-oct-2-ene 50 (100 mg, 0.33 mmol) was combined with ACE-Cl (0.25
mL) and the solution was heated to reflux for 5 h. The excess
chloroformate was removed in vacuo and the residue was refluxed in
methanol for 45 min. The methanol was removed in vacuo and the
residue was dissolved in CH.sub.2Cl.sub.2 and shaken with
NaHCO.sub.3/Na.sub.2CO.sub.3 (pH=9). The aqueous layer was
extracted CH.sub.2Cl.sub.2 (4.times.10 mL) and the combined organic
extracts were dried (Na.sub.2SO.sub.4), filtered and concentrated.
The residue was chromatographed (1-10% Et.sub.3N/EtOAc). Like
fractions were combined to yield compound 51 (27 mg, 28%).
[0213] R.sub.f0.15 (10% Et.sub.3N/EtOAc); .sup.1H-NMR (CDCl.sub.3)
.delta. 1.6-2.5 (m, 5H), 2.7-3.1 (m, 1H), 3.45 (s, 3H), 3.8-3.9 (m,
1H), 4.2-4.35 (m, 1H), 7.1-8.0 (m, 7H).
EXAMPLE 59
N-[2-(3'-N'-Propyl-(1''R)-2''-methoxycarbonyl-3''-(2-naphthyl)-trop-2-ene)-
)((2-((triphenylmethyl)
thio)ethyl)amino)acetyl]-S-(triphenyl)-2-aminoethanethiol (Compound
52, FIG. 10)
[0214] To a solution of
(1R)-2-methoxycarbonyl-3-(2-naphthyl)-8-azabicyclo[3.2.1]-oct-2-ene
51 (27 mg, 0.09 mmol) in dry CH.sub.3CN (10 mL) was added in
succession N-[[[2-[2-(triphenylmethyl)
thio]ethyl](N'-3'-chloropropyl)amino]acetyl]-S-(triphenylmethyl)-2-aminoe-
thanethiol (70 mg), KI (15 mg), and K.sub.2CO.sub.3 (127 mg). The
resulting slurry was maintained at reflux overnight. Once the
reaction was complete, the solution was allowed to cool to room
temperature and partitioned between conc. aq. NH.sub.4OH and
CH.sub.2Cl.sub.2. The layers were separated and the organic phase
dried with Na.sub.2SO.sub.4. The solution was filtered and
concentrated and the residue purified by flash chromatography
(50-90% EtOAc/hexanes+3% NH.sub.4OH) which gave 27 mg (29%) of
52.
[0215] R.sub.f 0.44 (1% NH.sub.4OH in EtOAc); .sup.1H-NMR
(CDCl.sub.3) .delta. 1.5-3.2 (m, 20H), 2.87 (s, 2H), 3.3-3.4 (m,
1H), 3.4 (s, 3H), 3.9-4.0 (m, 1H), 7.0-7.8 (m, 37H).
EXAMPLE 60
N-[(2-((3'-N'-Propyl-(1''R)-2''-methoxycarbonyl-3''-(2-naphthyl)trop-2-ene-
) (2-mercaptoethyl)amino)acetyl)-2-aminoethanethiolato]-rhenium (V)
oxide (Compound 41, FIG. 10) (O-1185)
[0216] A solution of
N-[2-(3'-N'-propyl-(1''])-2''-methoxycarbonyl-3''-(2-naphthyl)trop-2-ene--
))((2-((triphenylmethyl)
thio)ethyl)amino)acetyl]-S-(triphenyl)-2-aminoethanethiol 52 (27
mg, 0.027 mmol) in EtOH (10 mL) was heated to reflux. A solution of
SnCl.sub.2 (5.7 mg, 0.03 mmol) in 0.05 M HCl (1.0 mL) was added
quickly followed immediately by NaReO.sub.4 (8.2 mg, 0.03 mmol) in
0.05 M HCl (0.5 mL). The solution was refluxed for 10 h and was
then applied to a chromatography column (5% Et.sub.3N in
Et.sub.2O). Like fractions were combined and concentrated to yield
a mixture of the diastereomers (15 mg; 76%).
[0217] R.sub.f 0.15 (10% Et.sub.3N in EtOAc); IR (KBr) 967,
cm.sup.-1; Accurate mass calculated: 728.1599, found 728.1664;
.sup.1H-NMR (CDCl.sub.3) .delta. 1.3-4.3 (m, 23H), 3.45 (s, 3H),
4.5-4.9 (m, 2H), 7.1-8.0 (m, 7H).
EXAMPLE 61
(1R)-N-Methyl-2.beta.-methoxycarbonyl-3.beta.-(2-naphthyl)-8-azabicyclo[3.-
2.1]octane (Compound 42, FIG. 10) (O-1229) and
(1R)-N-Methyl-2.beta.-methoxycarbonyl-3.alpha.-(2-naphthyl)-8-azabicyclo[3-
.2.1]octane (Compound 43, FIG. 10) (O-1228)
[0218] To
(1R)-N-Methyl-2.beta.-methoxycarbonyl-3-(2-naphthyl)-8-azabicyc-
lo[3.2.1]-oct-2-ene 50 (510 mg, 1.66 mmol) in THF (15 mL) at
-78.degree. C. was added SmI.sub.2 solution (0.1 M in THF, 116 mL,
11.6 mmol) dropwise. After 30 min at -78.degree. C. MeOH (42 mL)
was added and the resulting solution stirred at -78.degree. C. for
a further hour. The reaction was then quenched by adding TFA and
water, the cold bath was also removed and the solution allowed to
attain room temperature. The reaction was then made basic with
NH.sub.4OH and diluted with ether and then filtered through celite.
The filter cake was washed with more ether and then all the organic
phases were combined and washed with a sodium thiosulfate solution
and then a brine solution. After drying with sodium sulfate the
solution was filtered and concentrated and gave the crude products
which were isolated by column chromatography (0-2% Et.sub.3N in
EtOAc). Compound 42 was isolated as a light yellow solid (110 mg,
22%).
[0219] Mp. 94-95.degree. C. R.sub.f 0.28 (10% MeOH/CHCl.sub.3); IR
(KBr) 2900, 1750 cm.sup.-1; .sup.1H-NMR (CDCl.sub.3) .delta.
1.5-2.3 (m, 5H), 2.25 (s, 3H), 2.7 (ddd, 1H), 2.9-3.3 (m, 2H), 3.45
(s, 3H), 3.3-3.4 (m, 1H), 4.5-4.6 (m, 1H), 7.3-7.5 (m, 3H), 7.7-7.9
(m, 4H). Elemental analysis: calculated (0.25 H.sub.2O) C, 76.52;
H, 7.55; N, 4.46; found C, 76.63; H, 7.57; N, 4.44.
[0220] Compound 43 was isolated as an off-white solid (113 mg,
23%).
[0221] M.p. 113-114.degree. C., R.sub.f 0.56 (10% MeOH/CHCl.sub.3);
IR (KBr) 3000, 1750 cm.sup.-1; .sup.1H NMR (CDCl.sub.3) .delta.
1.4-1.6 (ddd, 1H), 1.4-1.8 (m, 3H), 2.0-2.4 (m, 2H), 2.3 (s, 3H),
2.4-2.7 (m, 1H), 2.65 (dd, 1H), 3.2-3.4 (m, 3H), 3.57 (s, 3H),
7.2-7.5 (m, 3H). Elemental analysis: calculated C, 77.64; H, 7.49;
N, 4.53; found C, 77.48; H, 7.50; N, 4.45.
EXAMPLE 62
(1R)-2.beta.-Methoxycarbonyl-3.beta.-(.sup.2-naphthyl)-8-azabicyclo[3.2.1]-
-octane (Compound 44, FIG. 10)
[0222]
(1R)-N-Methyl-2.beta.-methoxycarbonyl-3.beta.-(2-naphthyl)-8-azabi-
cyclo[3.2.1]octane 42 (146 mg) was combined with ACE-Cl (5.5 mL)
and the solution was heated to reflux for 5 h. The excess
chloroformate was removed in vacuo and the residue was refluxed in
methanol for 45 min. The methanol was removed in vacuo and the
residue was dissolved in CH.sub.2Cl.sub.2 and shaken with
NaHCO.sub.3/Na.sub.2CO.sub.3 (pH=9). The aqueous layer was
extracted CH.sub.2Cl.sub.2 and the combined organic extracts were
dried (Na.sub.2SO.sub.4), filtered and concentrated. The residue
was chromatographed (2% MeOH/EtOAc). Like fractions were combined
to yield 44 (109 mg, 78%).
[0223] R.sub.f 0.27 (10% MeOH/EtOAc); .sup.1H-NMR (CDCl.sub.3)
.delta. 1.5-2.4 (m, 5H), 2.68 (ddd, 1H),4.8-4.9 (m, 1H), 3.30 (s,
3H), 3.3-3.4 (m, 1H), 3.7-3.9 (m, 2H), 7.1-8.0 (m, 7H). Elemental
analysis: calculated (0.25 H.sub.2O) C, 76.10; H, 7.23; N, 4.67;
found C, 75.98; H, 7.23; N, 4.60.
EXAMPLE 63
(1R)-2.beta.-Methoxycarbonyl-3.alpha.-(2-naphthyl)-8-azabicyclo[3.2.1]-oct-
ane (Compound 45, FIG. 10)
[0224]
(1R)-N-Methyl-2.beta.-methoxycarbonyl-3.alpha.-(2-naphthyl)-8-azab-
icyclo[3.2.1]-octane, 43 (90 mg, 0.29 mmol) was combined with
ACE-Cl (4 mL) and the solution was heated to reflux for 5 h. The
excess chloroformate was removed in vacuo and the residue was
refluxed in methanol for 45 min. The methanol was removed in vacuo
and the residue was dissolved in CH.sub.2Cl and shaken with
NaHCO.sub.3/Na.sub.2CO.sub.3 (pH=9). The aqueous layer was
extracted CH.sub.2Cl and the combined organic extracts were dried
(Na.sub.2SO.sub.4), filtered and concentrated. The residue was
chromatographed (2% MeOH/CHCl.sub.3). Like fractions were combined
to yield 45 (109 mg, 78%).
[0225] R.sub.f 0.29 (10% MeOH/CHCl.sub.3).
EXAMPLE 64
N-[2-(3'-N'-Propyl-(1''R)-2''.beta.-methoxycarbonyl-3''.beta.-(2-naphthyl)-
tropane)) ((2-((triphenylmethyl)
thio)ethyl)amino)acetyl]-S-(triphenyl)-2-aminoethanethiol (Compound
46, FIG. 10)
[0226] To a solution of
(1R)-2.beta.-methoxycarbonyl-3.beta.-(2-naphthyl)-8-azabicyclo[3.2.1]octa-
ne 44 (81 mg, 0.27 mmol) in dry CH.sub.3CN (20 mL) was added in
succession
N-[[[2-[2-(triphenylmethyl)thio]ethyl](N'-3'-chloropropyl)amino]acetyl]-S-
-(triphenylmethyl)-2-aminoethanethiol (207 mg), KI (46 mg, 0.27
mmol), and K.sub.2CO.sub.3 (378 mg). The resulting slurry was then
maintained at reflux overnight. Once the reaction was complete, the
solution was allowed to cool to room temperature and then
partitioned between concentrated aqueous NH.sub.4OH and
CH.sub.2Cl.sub.2. The layers were separated and the organic phase
dried with Na.sub.2SO.sub.4. The solution was filtered and
concentrated and the residue purified by flash chromatography
(0-90% EtOAc/hexanes+3% NH.sub.4OH) which gave 150 mg (54%) of
46.
[0227] R.sub.f 0.36 (10% Et.sub.3N in EtOAc); .sup.1H-NMR
(CDCl.sub.3) .delta. 1.4-3.8 (m, 24H), 2.85 (s, 2H), 3.38 (s, 3H),
7.0-7.8 (m, 37H).
EXAMPLE 65
N-[2-(3'-N'-Propyl-(1''R)-2''.beta.-methoxycarbonyl-3''.alpha.-(2-naphthyl-
)tropane)) ((2-((triphenylmethyl)
thio)ethyl)amino)acetyl]-S-(triphenyl)-2-aminoethanethiol (Compound
47, FIG. 10)
[0228] To a solution of
(1R)-2.beta.-methoxycarbonyl-3.alpha.-(2-naphthyl)-8-azabicyclo[3.2.1]oct-
ane 45 (17 mg, 0.06 mmol) in dry CH.sub.3CN (10 mL) was added in
succession N-[[[2-[2-(triphenylmethyl) thiol
ethyl](N'-3'-chloropropyl)amino]acetyl]-S-(triphenylmethyl)-2-aminoethane-
thiol (4.3 mg), KI (9.4 mg), and K.sub.2CO.sub.3 (79 mg). The
resulting slurry was then maintained at reflux overnight. Once the
reaction was complete, the solution was allowed to cool to room
temperature and then partitioned between concentrated aqueous
NH.sub.4OH and CH.sub.2Cl.sub.2. The layers were separated and the
organic phase dried with Na.sub.2SO.sub.4. The solution was
filtered and concentrated and the residue purified by flash
chromatography (50-80% EtOAc/hexanes+3% NH.sub.4OH) which gave 18
mg (31%) of 47.
[0229] R.sub.f 0.16 (50% EtOAc), 47% hexane, 3% NH.sub.4OH).
EXAMPLE 66
N-[(2-((3'-N'-Propyl-(1''R)-2''.beta.-methoxycarbonyl-3''.beta.-(2-naphthy-
l)-tropane)
(2-mercaptoethyl)amino)acetyl)-2-aminoethanethiolato]-rhenium (V)
oxide (Compound 48, FIG. 10) (O-1339)
[0230] A solution of
N-[2-(3'-N'-propyl-(1R)-2''.beta.-methoxycarbonyl-3''-(2-naphthyl)-trop-2-
-ene))((2-((triphenylmethyl)
thio)ethyl)amino)acetyl]-S-(triphenyl)-2-aminoethanethiol 46 (150
mg, 0.15 mmol) in EtOH (10 mL) was heated to reflux. A solution of
SnCl.sub.2 (31 mg, 0.16 mmol) in 0.05.M HCl (1.0 mL) was added
quickly followed immediately by NaReO.sub.4 (45 mg, 0.16 mmol) in
0.05 M HCl (0.5 mL). The solution was refluxed for 10 h and was
then applied to a chromatography column (5% Et.sub.3N in EtOAc).
Like fractions were combined and concentrated to yield a mixture of
the diastereomers (34 mg; 41%).
[0231] R.sub.f 0.39 (10% Et.sub.3N in EtOAc); .sup.1H-NMR
(CDCl.sub.3) .delta. 1.4-4.2 (m, 24H), 3.40, 3.48 (2s, 3H), 4.4-4.7
(m, 1H), 8.25, 8.30 (2d, 1H), 7.2-8.0 (m, 7H). Elemental analysis:
calculated (0.5 EtOAc) C, 46.97; H, 5.30; N, 5.39; found C, 46.91;
H, 5.12; N, 5.19.
EXAMPLE 67
N-[(2-((3'-N'-Propyl-(1''R)-2''.beta.-methoxycarbonyl-3''.alpha.-(2-naphth-
yl)-tropane)
(2-mercaptoethyl)amino)acetyl)-2-aminoethanethiolato]-rhenium (V)
oxide (Compound 49, FIG. 10)
[0232] A solution of
N-[2-(3'-N'-propyl-(1''R)-2''.beta.-methoxycarbonyl-3''.alpha.-(2-naphthy-
l)-trop-2-ene))((2-((triphenylmethyl)
thio)ethyl)amino)acetyl]-S-(triphenyl)-2-amino-ethanethiol 47 (18
mg, 0.02 mmol) in EtOH (10 mL) was heated to reflux. A solution of
SnCl.sub.2 (5.3 mg) in 0.05 M HCl (1.0 mL) was added quickly
followed immediately by NaReO.sub.4 (3.7 mg) in 0.05 M HCl (0.5
mL). The solution was refluxed for 10 h and was then applied to a
chromatography column (5% Et.sub.3N in Et.sub.2O). Like fractions
were combined and concentrated to yield a mixture of the
diastereomers (40%).
[0233] R.sub.f 0.46 (5% Et.sub.3N in EtOAc); .sup.1H-NMR
(CDCl.sub.3) .delta. 1.4-1.8 (m, 24H), 1.8-2.0 (m, 2H), 2.02-2.20
(m, 1H), 2.3-2.5 (m, 3H), 2.69, 2.71 (2s, 3H), 2.9-3.0 (2dd, 1H),
3.1-3.4 (m, 3H), 3.4-3.6 (m, 1H), 3.5,3.55 (2s, 3H), 3.6-3.8 (2ddd,
1H), 3.9-4.2 (m, 3H), 4.5-4.7 (m, 1H), 4.75, 4.98 (2d, 1H), 7.32
(d, 1H), 7.3-7.5 (m, 2H), 7.64 (s, 1H), 7.7-7.8 (m, 3H).
EXAMPLES 68-80
[0234] Tests were conducted to determine the binding affinity and
selectivity of certain compounds for the dopamine transporter. The
results are tabulated below. TABLE-US-00001 ##STR5## ##STR6##
##STR7## ##STR8## ##STR9## ##STR10## ##STR11## ##STR12## ##STR13##
##STR14## ##STR15## ##STR16## ##STR17## IC50 IC50 DAT/ Example
Compound DAT SERT SERT 68 O-1136 60 1693 28 69 O-861 2.9 80 28 70
O-863 40 -- -- 71 O-927 5.0 200 40 72 O-928 3.0 30 10 73 O-1185 63
960 15 74 O-1186 3.8 1300 340 75 O-1196 10 640 64 76 O-1339 3.6 59
16 77 O-1451 40 3260 80 78 O-1505 2.0 497 249 79 O-1508 5.9 200 34
80 O-1561 5.3 337 63
EXAMPLES 81-91
.sup.99mTc Labeling
[0235] The following compounds were prepared by identical methods
described below.
[0236] 81
N-[(2-((3'-N'-Propyl-(1''R)-2''.beta.-methoxycarbonyl-3''.alpha-
.-(3,4-dichlorophenyl)-tropane)-(2-mercaptoethyl)amino)acetyl)-2-aminoetha-
nethiolato]-technetium (V) oxide (Compound 37 (R=3,4-Cl.sub.2),
FIG. 8) (O-1196)
[0237] 82
N-[(2-((3'-N'-Propyl-(1''R)-2''.beta.-1-propanoyl-3''.alpha.-(4-
-fluorophenyl)-tropane)-(2-mercaptoethyl)amino)acetyl)-2-aminoethanethiola-
to]technetium (V) oxide (Compound 19 (R=4-F), FIG. 4)
[0238] 83
N-[(2-((3'-N'-Propyl-(1''R)-2''.beta.-1-propanoyl-3''.alpha.-(3-
,4-dichlorophenyl)-tropane)
(2-mercaptoethyl)amino)acetyl)-2-aminoethanethiolato]-technetium
(V) oxide (Compound 19 (R=3,4-Cl.sub.2), FIG. 4)
[0239] 84
N-[(2-((3'-N'-Propyl-(1''R)-2''.beta.-methoxymethylcarbamoyl-3'-
'.alpha.-(4-fluorophenyl)-tropane)
(2-mercaptoethyl)amino)acetyl)-2-aminoethanethiolato]-technetium
(V) oxide (Compound 22 (R=4-F), FIG. 5) (O-1451)
[0240] 85
(RS)-N-[2-((3-N'-Propyl-(1''R-2''.beta.-carbomethoxy-3''.beta.--
(4-fluorophenyl)-tropane))
(2-mercaptoethyl)amino)acetyl)-2-aminoethanethiolato]-technetium
(V) oxide (Compound 34 (R=4-F), FIG. 8)
[0241] 86
(RS)-N-[2-((3-N'-Propyl-(1''R-2''.beta.-carbomethoxy-3''.beta.--
(3,4-dichlorophenyl)-tropane))
(2-mercaptoethyl)amino)acetyl)-2-aminoethanethiolato]-technetium
(V) oxide (Compound 34 (R=3,4-Cl.sub.2), FIG. 8)
[0242] 87
N-[(2-((3'-N'-Propyl-(1''R)-2''-carbomethoxy-3''-(4-fluoropheny-
l)-tropene)
(2-mercaptoethyl)amino)acetyl)-2-aminoethanethiolato]-technetium
(V) oxide (Compound 31 (R=4-F), FIG. 7)
[0243] 88
N-[(2-((3'-N'-Propyl-(1''R)-2''-methoxycarbonyl-3''-(3,4-dichlo-
rophenyl)-tropene)
(2-mercaptoethyl)amino)acetyl)-2-aminoethanethiolato]-technetium
(V) oxide (Compound 31 (R=3,4-Cl.sub.2), FIG. 7)
[0244] 89
N-[(2-((3'-N'-Propyl-(1''R)-2''.beta.-methoxycarbonyl-3''.alpha-
.-(4-fluorophenyl)-tropane)
(2-mercaptoethyl)amino)acetyl)-2-aminoethanethiolato]-technetium
(V) oxide (Compound 37 (R=4-F), FIG. 8)
[0245] 90
N-[(2-((3'-N'-Propyl-(1''R)-3''.beta.-(4-fluorophenyl)tropane-2-
''.beta.-1-propanoyl)-(2-mercaptoethyl)amino)acetyl)-2-aminoethanethiolato-
]technetium (V) oxide (Compound 11 (R=4-F), FIG. 3)
[0246] 91
N-[(2-((3'-N'-Propyl-(1''R)-2''.beta.-1-propanoyl-3''.beta.-(3,-
4-dichlorophenyl)-tropane)
(2-mercaptoethyl)amino)acetyl)-2-aminoethanethiolato]-technetium
(V) oxide (Compound 11 (R=3,4-Cl.sub.2), FIG. 3)
Deprotection of Tropane Analog for Radiolabeling:
[0247] The following is a description of the procedures that were
employed:
[0248] Twenty microliter aliquots of TFA, CH.sub.2Cl.sub.2, and
(C.sub.2H.sub.5).sub.3SiH were added to 5.0 mg of each trityl
protected precursor to cleave the thiol protecting group. After a
20 min. incubation, 200 L of 1.0 M HCl/Ether was added to protonate
the thiols. The solvent was evaporated off, followed by successive
washes with hexanes to remove the trityl groups. The deprotected
compound was then dissolved in DMSO to produce a stock solution at
a concentration of 1 mg/mL.
Radiolabeling the Tropane Analog:
[0249] Approximately 200 mCi of sodium .sup.99mTc pertechnetate was
added to a Glucoheptonate kit (Du Pont Pharmaceuticals, Billerica,
Mass.) and allowed to incubate at room temperature for 15 min. 150
mCi of the resulting .sup.99mTc-Glucoheptonate was added to an
equal volume of 50 mM Acetate Buffer, pH 5.2 (approximately 2 mL)
and 50 .mu.L of deprotected precursor stock solution (1 mg/mL in
DMSO, 50 .mu.g). This solution was incubated at room temperature
for 20 min. The course of the radiolabeling was monitored with a
Rainin HPLC system using a reverse phase Vydac C8 column
(4.6.times.250 mm 5 .mu.m). The column was eluted with a 0.1M
ammonium acetate/acetonitrile mobile phase; 1.5 mL/min. flow rate,
and a gradient of 5-100% acetonitrile over 15 minutes. Radioactive
detection is achieved with a Frisk - Tech Rate meter (Bicron
Corp.). The final radiolabeled product was purified using a C18
Sep-Pak (Waters Inc.) eluted with ethanol. For all compounds,
labeling yields and radiochemical purities were greater than 85%
and 98%, respectively. Each product was diluted with sterile saline
to yield a <10% ethanol solution followed by filtration through
a 0.22 .mu.m filter prior to injection. Typical HPLC chromatograms
are shown in FIGS. 11-14.
Summary of analytical results, HPLC:
[0250] With the above methodology, it was observed that the
retention times were dependent upon two features of the analog,
i.e., the conformation and the substituents. As expected the
dichloro-analogs eluted later than the monofluorinated compounds
and the boat conformers eluted later than the respective chair
compounds. Retention times for four specific .sup.99mTc labeled
compounds are tabulated below. The reproducibility was .+-.0.1 min.
TABLE-US-00002 Example No. Compound Retention Time(min.) Example 82
0-1505T 16.6 Example 90 0-1508T 15.2 Example 83 0-1561T 17.1
Example 91 0-1506T 16.1
Animal Model of Parkinson's Disease:
[0251] The neurotoxin MPTP (N-methyl-1,2,3,6-tetrahydropyridine),
when administered to monkeys, produces a spectrum of motor,
cognitive, biochemical and morphological changes that is not
replicated in rodents. MPTP-treated monkeys develop neurological
deficits (resting tremor, rigidity, akinesia, postural
abnormalities), morphological changes (cell loss in the substantia
nigra, ventral tegmental area, retrorubal fields) and biochemical
changes (severe depletion of DA and decreases in norepinephrine and
serotonin) that closely, parallel idiopathic PD and
post-encephalitic Parkinsonism.
[0252] Two monkeys were treated with MPTP; 3-5 doses of 0.6 mg/kg
administered over 10 days. Treatment was performed under ketamine
anesthesia (15 mg/kg). This dose of MPTP, has previously been used
to produce Parkinsonism within 2-3 weeks and depletion of .sup.3H
or .sup.11C CFT binding sites by one month. Animals treated in this
manner were expected to show an inverse relationship between SPECT
tracer binding and motor dysfunction. The marked depletion of DA
terminals produced by this treatment should provide definitive
evidence of the selectivity and sensitivity of the SPECT tracers to
detect full depletion of the nerve terminals.
SPECT Imaging
[0253] All SPECT images were acquired with a MultiSPECT 2 gamma
camera (Siemens, Hoffman Estates, IL) equipped with fan-beam
collimators and peaked to the 140 KeV photopeak of .sup.99mTc (15%
window). This camera has intrinsic resolution of 4.6 mm (FWHM), and
a sensitivity of .about.240 cps/mCi. Images were acquired over
360.degree. (60 projections/head, 128.times.128 matrix) in the
continuous mode. Image reconstruction was performed using a
conventional filtered back-projection algorithm to an in-plane
resolution of 10 mm FWHM and attenuation correction via the Chang
method.
[0254] Rhesus monkeys weighing approximately 7 kg were anesthetized
with ketamine/xylazine (15.0 and 1.5 mg/kg) and positioned prone on
the imaging bed of the SPECT camera. Before the start of imaging, a
venous catheter was inserted in a peripheral vein for
radiopharmaceutical administration. The heads of the animals were
immobilized with a custom fabricated head holder. Approximately
20-25 mCi of .sup.99mTc labeled O-1505T, O-1508T, O-1561T or
O-1560T was injected intravenously over 60 seconds. Dynamic SPECT
imaging was initiated at the end of the infusion and consisted of 2
min. acquisitions during the first hour and 5 min. acquisitions
thereafter.
Selectivity of for DAT Sites in the Monkey Brain
[0255] For this study, a monkey was positioned supine on the
imaging table of the MultiSPECT 2 gamma camera and injected with
.about.20 mCi of .sup.99mTc O-1560T as described above. At
approximately 20 min. after radiopharmaceutical administration, the
animal was injected with CFT (1.0 mg/kg) and imaging was continued
for an additional 70 min.
Image Analysis
[0256] SPECT slices with greatest striatal activity or in which the
occipital cortex was well visualized were summed and regions of
interest (ROI's) were constructed. In the striatal planes, ROI's
were placed on the right and left striatum (caudate +putamen).
[0257] Radioactivity in the right and left striatum were averaged.
ROI radioactivities (cpm/cc) were decay corrected to the time of
injection. The difference between striatal and occipital cortex
activity (specific binding) was calculated for each image and was
plotted as a function of time. Corrections for scattered fraction
and partial volume effects were not performed.
[0258] Radiation Dosimetry
[0259] Groups of male Sprague-Dawley rats were in injected with
.about.10 mCi of .sup.99mTc labeled O-1505T and O-1561T. At time
intervals between 5 min. and 24 hrs after injection, groups of 6
animals injected with each radiopharmaceutical were sacrificed and
biodistribution was measured. Samples of blood, heart, lung, liver,
spleen, kidney, adrenal, stomach, GI tract, testes, bone, bone
marrow and skeletal muscle were weighed and radioactivity was
measured with a well type gamma counter (LKB model # 1282, Wallac
Oy, Finland). To correct for radioactive decay and permit
calculation of the concentration of radioactivity in each organ as
a fraction of the administered dose, aliquots of the injected doses
were counted simultaneously. The results were expressed as percent
injected dose per gram (% I.D./g) and radiation dosimetry was
estimated by the MIRdose method.
RESULTS
SPECT Imaging with 99mTc Labeled O-1505T, O-1508T, O-1561 T and
O-1560T
[0260] In the images acquired early after injection of .sup.99mTc
O-1505T and .sup.99mTc O-1561T, diffuse accumulation of
radioactivity was observed throughout the brain. Over the first
several minutes after injection, accumulation of tracer in the
striatum intensified and the level of radioactivity in all other
structures decreased. By 30 minutes after injection there was
excellent contrast between striatum and the rest of the brain.
Trans-axial, sagittal and coronal images of monkey brains were
acquired between 30 and 50 minutes after injection of .sup.99mTc
labeled O-1505T, O-1508T, O-1561T and O-1560T. From these data, it
is clear that the images acquired after injection of .sup.99mTc
0-1505T and .sup.99mTc O-1561T (boat forms of the monofluoro and
dichloro compounds, respectively) display high concentrations of
radiopharmaceutical in the striatum with minimal accumulation in
other areas of the brain. In particular, lack of accumulation in
the thalamus, hypothalamus or midbrain, regions that are rich in
5-HT transporters, supports the specificity of this tracer for DAT
sites. Region of interest analysis yielded striatal to occipital
cortex ratios of approximately .about.2.5 to 1. In this experiment,
images acquired after injection of .sup.99mTc O-1508T and
.sup.99mTc O-1560T (chair forms of the monofluoro and dichloro
compounds, respectively) differed with O-1508T failing to
demonstrate significant accumulation of radioactivity in striatum
or elsewhere in the brain while O-1560T provided results more like
O-1505T and O-1561T.
SPECT of MPTP Treated Monkeys
[0261] (Mid-striatal) trans-axial, sagittal and coronal SPECT
images of the brain of a rhesus monkey injected with .sup.99mTc
O-1505T were obtained one month after MPTP treatment. Compared with
normal animals, after MPTP treatment, the level of accumulation
decreased markedly and the striatum could not be differentiated
from surrounding structures.
[0262] Selectivity of 99mTc 0-1505T for DAT sites in monkey brain
SPECT images of monkey brain showed significant accumulation of
.sup.99mTc O-1505T in the striatum. In the early images there was
diffuse accumulation of radioactivity throughout the brain. Over
the first several minutes after injection, accumulation of tracer
in the striatum intensified and the level of radioactivity in all
other structures decreased. By 30 minutes after injection there was
excellent contrast between striatum and the rest of the brain.
After injection of a receptor saturating dose of unlabeled CFT,
striatal accumulation of radioactivity decreased and by 60 minutes
after injection, there was no evidence of focal accumulation in the
striatum.
Radiation Dosimetry
[0263] The biodistribution studies demonstrated that both
.sup.99mTc -1505T and .sup.99mTc O-1561 cleared rapidly from all
tissues of the rat. For .sup.99mTc O-1505T, MIRdose calculations
revealed urinary bladder to be the target organ with a dose of 0.29
rem/mCi. Total body effective dose was estimated at 0.037
rem/mCi
Summary:
[0264] These results demonstrate that .sup.99mTc labeled O-1505T
and O-1561T are excellent SPECT ligands for DAT sites. These
radiopharmaceuticals combine the the following important
characteristics for obtaining useful diagnostic images: (1) high
striatal to occipital cortex ratios; (2) high selectivity for DAT
vs. 5-HT transporter (SET) sites; (3) convenient preparation at
high specific activity and radiochemical purity; (4) favorable
radiation dosimetry and (5) striatal localization rate that is well
matched to the physical t1/2 of .sup.99mTc.
[0265] The present invention has been described in detail,
including the preferred embodiments thereof. However, it will be
appreciated that those skilled in the art, upon consideration of
the present disclosure, may make modifications and/or improvements
of this invention and still be within the scope and spirit of this
invention as set forth in the following claims:
* * * * *