U.S. patent application number 14/223782 was filed with the patent office on 2014-11-20 for beta-amyloid pet imaging agents.
This patent application is currently assigned to The United States of America, as represented by the Secretary, Department of Health and Human Serv. The applicant listed for this patent is The United States of America, as represented by the Secretary, Department of Health and Human Serv, The United States of America, as represented by the Secretary, Department of Health and Human Serv. Invention is credited to Lisheng Cai, Robert B. Innis, Victor W. Pike.
Application Number | 20140341812 14/223782 |
Document ID | / |
Family ID | 38566135 |
Filed Date | 2014-11-20 |
United States Patent
Application |
20140341812 |
Kind Code |
A1 |
Cai; Lisheng ; et
al. |
November 20, 2014 |
BETA-AMYLOID PET IMAGING AGENTS
Abstract
Novel derivatives of imidazopyridinylbenzeneamines and novel
derivatives of benzothiazolylbenzeneamines are disclosed that offer
improved behavior when used as imaging agents for positron emission
tomography of beta-amyloids. Also disclosed is a
palladium-catalyzed reaction scheme under microwave conditions for
aryl thioethers in general that provides a high ratio of
substitution relative to reduction and can be used for the
imidazopyridinylbenzeneamine derivatives as well as other compounds
of related structure.
Inventors: |
Cai; Lisheng; (Derwood,
MD) ; Pike; Victor W.; (Bethesda, MD) ; Innis;
Robert B.; (Rockville, MD) |
|
Applicant: |
Name |
City |
State |
Country |
Type |
The United States of America, as represented by the Secretary,
Department of Health and Human Serv |
Bethesda |
MD |
US |
|
|
Assignee: |
The United States of America, as
represented by the Secretary, Department of Health and Human
Serv
Bethesda
MD
|
Family ID: |
38566135 |
Appl. No.: |
14/223782 |
Filed: |
March 24, 2014 |
Related U.S. Patent Documents
|
|
|
|
|
|
Application
Number |
Filing Date |
Patent Number |
|
|
12293340 |
Sep 22, 2010 |
8703096 |
|
|
PCT/US2007/066939 |
Apr 21, 2006 |
|
|
|
14223782 |
|
|
|
|
60793807 |
Apr 21, 2006 |
|
|
|
Current U.S.
Class: |
424/9.44 ;
548/178 |
Current CPC
Class: |
A61P 25/28 20180101;
A61K 51/00 20130101; C07D 277/66 20130101; C07D 471/04 20130101;
A61K 51/0459 20130101; A61K 49/04 20130101; A61K 49/0433 20130101;
A61K 51/0453 20130101; A61K 51/0455 20130101 |
Class at
Publication: |
424/9.44 ;
548/178 |
International
Class: |
A61K 49/04 20060101
A61K049/04 |
Claims
1-10. (canceled)
11. A compound having the formula ##STR00020## wherein: R.sup.11 is
a member selected from the group consisting of C.sub.1-C.sub.6
alkyl, aryl- or halo-substituted C.sub.1-C.sub.6 alkyl,
hydroxyl-substituted C.sub.1-C.sub.6 alkyl, and carbamoyl
C.sub.1-C.sub.6alkyl; R.sub.12 is a member selected from the group
consisting of H, C.sub.1-C.sub.6 alkyl, halo-substituted
C.sub.1-C.sub.6 alkyl, halo, and C.sub.1-C.sub.6 alkylthio, and
R.sup.13 is a member selected from the group consisting of H and
C.sub.1-C.sub.3 alkyl, or R.sup.12 and R.sup.13 are joined to form
a thio-C.sub.1-C.sub.3 alkylene or alkylene linkage, and R.sup.14
is a member selected from the group consisting of H and
C.sub.1-C.sub.6 alkyl, and X is CH or N.
12. The compound of claim 11 wherein R.sup.11 is a member selected
from the group consisting of C.sub.1-C.sub.3 alkyl,
fluoro-substituted C.sub.1-C.sub.3 alkyl, hydroxyl-substituted
C.sub.1-C.sub.3 alkyl, and carbamoyl C.sub.1-C.sub.3 alkyl.
13. The compound of claim 11 wherein R.sup.11 is a member selected
from the group consisting of methyl, ethyl, fluoromethyl,
fluoroethyl, hydroxymethyl, hydroxyethyl, carbamoylmethyl, and
carbamoylethyl.
14. The compound of claim 11 wherein R.sup.12 is a member selected
from the group consisting of H, C.sub.1-C.sub.3 alkyl, chloro,
bromo, chloro-substituted C.sub.1-C.sub.3alkyl, bromo-substituted
C.sub.1-C.sub.3 alkyl, and C.sub.1-C.sub.3 alkylthio, and R.sup.13
is a member selected from the group consisting of H and
C.sub.1-C.sub.3 alkyl, or R.sup.12 and R.sup.13 are joined to form
a thiomethylene or alkylene linkage.
15. The compound of claim 11 wherein R.sup.12 is a member selected
from the group consisting of H, bromo, and methylthio, and R.sup.13
is a member selected from the group consisting of H and methyl, or
R.sup.12 and R.sup.13 are joined to form a thiomethylene or
alkylene linkage.
16. The compound of claim 11 wherein R.sup.14 is a member selected
from the group consisting of H and methyl.
17. The compound of claim 11 wherein R.sup.11 is a member selected
from the group consisting of C.sub.1-C.sub.3 alkyl,
fluoro-substituted C.sub.1-C.sub.3 alkyl, hydroxyl-substituted
C.sub.1-C.sub.3 alkyl, and carbamoyl C.sub.1-C.sub.3 alkyl;
R.sup.12 is a member selected from the group consisting of H,
C.sub.1-C.sub.3 alkyl, chloro, bromo, chloro-substituted
C.sub.1-C.sub.3 alkyl, bromo-substituted C.sub.1-C.sub.3 alkyl, and
C.sub.1-C.sub.3 alkylthio, and R.sup.13 is a member selected from
the group consisting of H and C.sub.1-C.sub.3 alkyl, or R.sup.12
and R.sup.13 are joined to form a thiomethylene or alkylene
linkage; and R.sup.14 is a member selected from the group
consisting of H and methyl.
18. A method for imaging amyloid deposits in a patient, said method
comprising administering to said patient a compound of claim 11 and
imaging portions of said patient where amyloid deposits occur when
said patient is suffering from Alzheimer's disease.
19. A method for imaging amyloid deposits in a patient, said method
comprising administering to said patient a compound of claim 11 and
imaging portions of said patient where amyloid deposits occur when
said patient is suffering from Alzheimer's disease.
Description
CROSS REFERENCE TO RELATED APPLICATION
[0001] This Application is a Divisional of U.S. patent application
Ser. No. 12/293,340, filed Sep. 22, 2010, which is a U.S. National
Stage Application of International Application No.
PCT/US2007/066939, filed Apr. 19, 2007, which claims priority from
U.S. Provisional Patent Application No. 60/793,807, filed Apr. 21,
2006 all of which are incorporated by reference in their
entirety.
BACKGROUND OF THE INVENTION
[0002] All publications cited herein, including patents and
published articles, are hereby incorporated herein by
reference.
[0003] Alzheimer's disease (AD) is the most common cause of
dementia and is characterized by progressive impairment in
cognitive function and behavior. This progressive, irreversible
brain disorder affects millions of lives and imposes a devastating
burden on the health care around the world. Over the past two
decades, significant progress has been made in deciphering the
pathogenesis and developing new therapeutic approaches. The
pathological features of AD include neuritic plaques composed of
amyloid-.beta. peptide (A.beta.) fibrils, neurofibrillary tangles
of hyper-phosphorylated tau, and neurotransmitter Deficits. Recent
efforts of managing AD have been focused on the prevention of
production, aggregation, and deposition of amyloid-.beta. peptides
in the brain and the acceleration of clearance of A.beta. from the
brain.
[0004] Non-invasive detection and quantitation of amyloid deposits
in the brain has been used to develop anti-amyloid therapies.
Direct imaging of amyloid load in vivo inpatients with AD is useful
for the early diagnosis of AD and the development and assessment of
treatment strategies. To this end, compounds suitable for in vivo
imaging of amyloid deposits in human brains have been developed.
Among these compounds are monoclonal antibodies against A.beta. and
peptide fragments, but these have had limited uptake by the brain
when tested in patients with AD, Putrescine-gadolinium-A.beta. has
been injected into transgenic mice overexpressing amyloid, and this
has resulted in labeling observed with MRI. Amyloid deposition can
also be non-invasively imaged and quantitated with a radiotracer
that readily enters the brain and selectively binds to amyloid
deposits.
[0005] The small molecule approach for amyloid imaging has so far
been the most Successful. Some of the promising compounds that have
been used to image amyloid are based on Congo red, thioflavin,
stilbene, and FDDNP. The binding of different derivatives of Congo
red and thioflavin has been studied in human autopsy brain tissue
and in transgenic Mice. Two compounds in advanced testing,
fluorine-18-labelled-FDDNP
(2-(1-{6-[(2-[.sup.18F]fluoroethyl)(methyl)amino]-2-naphthyl}ethylidene)m-
alononitrile) and carbon-11-labelled-PIB
(6-hydroxy-2-(4'-methylaminophenyl)benzothiazole; Pittsburg
compound B), both show more binding in the brains of patients with
AD than in those of healthy people. Table 1 below, which is taken
from Nordberg, A., "PET imaging of amyloid in Alzheimer's disease,"
The Lancet Neurology 3(9), September 2004, pp, 519-527, provides a
list of compounds presently undergoing evaluation as amyloid
imaging agents.
TABLE-US-00001 TABLE 1 Prior Art Compounds Used for Imaging of
Amyloid in the Brains of Living Organisms (reference numerals refer
to citations in the Nordberg paper) Imaging Reference Imaging
compound technique Study Friedland et al..sup.40 .sup.99Tc-10H3
SPECT Patients with AD Shoghi-Jadid et .sup.18F-FDDNP PET Patients
with AD al..sup.41 Klunk et al..sup.39 .sup.11C-PIB PET Patients
with AD Mathis et al..sup.42 .sup.11C-BTA-1 PET Baboons Poduslo et
al..sup.43 MION-A.beta..sub.1-40 MRI Mice PS Poduslo et al..sup.43
PUT-Gd-A.beta. MRI Mice PS Wadghiri et al..sup.44
Gd-DTPA-A.beta..sub.1-40 NMRI Mice APP/PS Mathis et al..sup.45
BTA-1 Multiphoton Mice APP/PS Bacskai et al..sup.46 Thioflavin-S
Multiphoton Mice APP Bacskai et al..sup.46 PIB Multiphoton Mice
APP
[0006] Two tertiary amines, [.sup.18F]FEM-IMPY
[N-(2-fluoroethyl)-4-(6-iodo-H-imidazo[1,2-A]pyridin-2-yl)-N-methylbenzen-
eamine], and its 3-fluoropropyl analog, [.sup.18F]FPM-IMPY, have
been previously evaluated by inventors herein as .beta.-amyloid
radioligands and reported in Cai, L., et al, "Synthesis and
evaluation of two .sup.18F-labeled
6-iodo-2-(4'-N,N-dimethylamino)phenylimidazo[1,2-a]pyridine
derivatives as prospective radioligands for beta-amyloid in
Alzheimer's disease," J. Med. Chem. 2004, 47, 2208-2218. After
intravenous injection of either radioligand into rodent or monkey
there is rapid and high uptake of radioactivity into brain with an
SUV (standardized uptake value, % I.D.kg/g) of about 160 followed
by biphasic clearance with a fast and a very slow component. These
radioligands were rapidly metabolized by processes involving
de-alkylation of the tertiary aromatic amino group, culminating in
defluoridation and high uptake of radioactivity in bone.
Tetra-deuteration of the fluoroethyl group did not lead to a
significant reduction in the residual brain radioactivity, but
reduced the bone uptake of radioactivity, presumably due to an
isotope effect on metabolism. An object of the present invention is
to provide radioligands that do not undergo rapid defluoridation
and do not produce residual radioactivity in the brain. The
invention therefore resides in further analogs of
imidazo[1,2-a]pyridine (IMPY) useful as radioligands for the
detection of A.beta. amyloid aggregates in Alzheimer's Disease (AD)
patients. Preferred among these analogs are imidazo[1,2-a]pyridine
N-methylbenzeneamines in which a methyl group or a group similar in
size to a methyl group (such as a bromine atom) is bonded to the
ortho ring position nearest to the amine.
[0007] The effective management of Alzheimer's disease requires
tools to diagnose, monitor, treat and prevent the disease. An
object of the present invention is therefore to provide amyloid
imaging agents with high specificity of binding to beta amyloid,
low background noise, better entry into brains, and improved
labeling efficiencies.
THE INVENTION
[0008] Two novel classes of amyloid imaging agents are disclosed
herein. The first class are derivatives of
imidazopyridinylbenzeneamine (IMPY), and are defined by Formula
(I):
##STR00001##
[0009] In Formula (I): [0010] R.sup.1 is C.sub.1-C.sub.6 alkyl, or
aryl-, or halo-substituted C.sub.1-C.sub.6 alkyl methyl; preferably
C.sub.1-C.sub.6 alkyl or fluoro-substituted C.sub.1-C.sub.6 alkyl;
and most preferably methyl, ethyl, fluoromethyl, or fluoroethyl,
[0011] R.sup.2 is H, C.sub.1-C.sub.6 alkyl, halo-substituted
C.sub.1-C.sub.6 alkyl, halo, or C.sub.1-C.sub.6 alkylthio, and
R.sup.3 is H or C.sub.1-C.sub.6 aklyl, or R.sup.2 and R.sup.3 are
joined to form a thio-C.sub.1-C.sub.6 allylene linkage (--S-alkyl-)
linkage or alkylene linkage between the 2-position (to which
R.sup.2 is attached in the formula) on the phenyl ring and the
amine nitrogen (to which R.sup.3 is attached) with the S atom
attached to the 2-position carbon; and preferably R.sup.2 is
hydrogen, bromo, or methylthio, R.sup.3 is hydrogen or methyl, or
R.sup.2 and R.sup.3 are joined to form a thiomethylene
(--SCH.sub.2--) linkage between the 2-position (to which R.sup.2 is
attached in the formula) on the phenyl ring and the amine nitrogen
(to which R.sup.3 is attached) with the S atom attached to the
2-position carbon, and [0012] R.sup.4 is H or C.sub.1-C.sub.3
alkyl, and R.sup.5 is H or C.sub.1-C.sub.3 alkyl, or R.sup.4 and
R.sup.5 are joined to form a C.sub.1-C.sub.3 alkylenethio linkage
(-alkyl--S--) linkage between the amine nitrogen (to which R.sup.4
is attached) and the 6-position (to which R.sup.3 is attached in
the formula) on the phenyl ring with the S atom attached to the
2-position carbon; and preferably R.sup.4 is H and R.sup.5 is H, or
R.sup.4 and R.sup.5 are joined to form a methylenethio
(--CH.sub.2S--) linkage between the amine nitrogen (to which
R.sup.4 is attached) and the 6-position (to which R.sup.5 is
attached in the formula) on the phenyl ring with the S atom
attached to the 2-position carbon.
TABLE-US-00002 [0012] TABLE 1 Examples of compounds within the
scope of Formula (I) are: Example R.sup.1 R.sup.2 R.sup.3 R.sup.4
R.sup.5 1.1 CH.sub.3 H CH.sub.3 H H 1.2 C.sub.2H.sub.5 H CH.sub.3 H
H 1.3 CH.sub.3 Br H H H 1.4 FCH.sub.2 H CH.sub.3 H H 1.5 FCH.sub.2
Br H H H 1.6 FCH.sub.2CH.sub.2 H CH.sub.3 H H 1.7 FCH.sub.2CH.sub.2
Br H H H 1.8 FCH.sub.2 SCH.sub.3 H H H 1.9 FCH.sub.2 SCH.sub.2 H H
1.10 FCH.sub.2 SCH.sub.2 SCH.sub.3
[0013] The second class are derivatives of
benzothiazolylbenzeneamine (BTA), and are defined by Formula
(II):
##STR00002##
[0014] In Formula (II); [0015] R.sup.11 is C.sub.1-C.sub.6 alkyl,
halo-substituted C.sub.1-C.sub.6 alkyl, hydroxy-substituted
C.sub.1-C.sub.6 alkyl, or carbamoyl C.sub.1-C.sub.6 alkyl;
preferably C.sub.1-C.sub.3 alkyl, fluoro-substituted
C.sub.1-C.sub.3alkyl, hydroxy-substituted C.sub.1-C.sub.3 alkyl, or
carbamoyl C.sub.1-C.sub.3 alkyl; and most preferably methyl, ethyl,
fluoromethyl, fluoroethyl, hydroxymethyl, hydroxyethyl,
carbamoylmethyl, or carbamoylethyl; [0016] R.sup.12 is H,
C.sub.1-C.sub.6 alkyl, halo-substituted C.sub.1-C.sub.6 alkyl,
halo, and C.sub.1-C.sub.6 alkylthio, and R.sup.13 is a member
selected from the group consisting of H and C.sub.1-C.sub.6 alkyl,
or R.sup.12 and R.sup.13 are joined to form a thio-C.sub.1-C.sub.3
alkylene linkage (--SCH.sub.2--) between the 2-position (to which
R.sup.12 is attached in the formula) on the phenyl ring and the
amine nitrogen (to which R.sup.13 is attached) with the S atom
attached to the 2-position carbon; preferably H, C.sub.1-C.sub.3
alkyl, chloro, bromo, chloro-substituted C.sub.1-C.sub.3 alkyl,
bromo-substituted C.sub.1-C.sub.3 alkyl, and C.sub.1-C.sub.3
alkylthio, and R.sup.13 is a member selected from the group
consisting of H and C.sub.1-C.sub.3 alkyl, or R.sup.12 and R.sup.13
are joined to form a thiomethylene linkage (--SCH.sub.2--) between
the 2-position (to which R.sup.12 is attached in the formula) on
the phenyl ring and the amine nitrogen (to which R.sup.13 is
attached) with the S atom attached to the 2-position carbon; and
most preferably R.sup.12 is hydrogen, bromo, or methylthio,
R.sup.13 is hydrogen or methyl, or R.sup.12 and R.sup.13 are joined
to form a thiomethylene (--SCH.sub.2--) linkage between the
2-position (to which R.sup.12 is attached in the formula) on the
phenyl ring and the amine nitrogen (to which R.sup.13 is attached)
with the S atom attached to the 2-position carbon, and [0017]
R.sup.14 is H or C.sub.1-C.sub.6 alkyl, preferably H or methyl, and
[0018] X is CH or N,
TABLE-US-00003 [0018] TABLE 2 Examples of compounds within the
scope of Formula (II) are: Example R.sup.11 R.sup.12 R.sup.13
R.sup.14 X 2.1 CH.sub.3 H CH.sub.3 H N 2.2 H.sub.2N--C(O)--CH.sub.2
H CH.sub.3 H CH 2.3 HOC.sub.2H.sub.4 Br H H CH 2.4 FCH.sub.2 H
CH.sub.3 H N 2.5 FCH.sub.2 Br H H N 2.6 FCH.sub.2CH.sub.2 H
CH.sub.3 H N 2.7 FCH.sub.2CH.sub.2 Br H H N 2.8 FCH.sub.2 SCH.sub.3
H H N 2.9 FCH.sub.2 SCH.sub.2 H N 2.10 FCH.sub.2 SCH.sub.2 CH.sub.3
N
[0019] This invention resides in the above compounds as novel
imaging agents for the imaging of amyloid deposits, new methods of
synthesis of imidazopyridinylbenzeneamine (IMPY) derivatives, and
methods for diagnosing Alzheimer's disease in vivo by positron
emission tomography, magnetic resonance imaging and other imaging
methods involving the use of the imaging agents of this
invention,
BRIEF DESCRIPTION OF THE DRAWINGS
[0020] FIG. 1. Synthesis of ketones and .alpha.-bromoketones as
intermediates in the preparation of IMPY derivatives.
[0021] FIG. 2. Synthesis of 2-aminopyridine derivatives and
2-aminopyrazines. The synthetic methods of pyridine derivatives
depend on the substituents on the ring.
[0022] FIG. 3. Aromatic substitutions with the methoxy anion. The
method uses salts of copper (I) and a small amount of ester to form
a stabilized tetrahedral adduct.
[0023] FIG. 4. Synthesis of 5-chloro-2-aminopyrazine using mild
conditions,
[0024] FIG. 5. Synthsesis of IMPY Derivatives I. Most of the IMPY
derivatives were synthesized through the direct condensation of
.alpha.-bromoketones and 2-amino-pyridines.
[0025] FIG. 6. Synthesis of IMPY Derivatives II using direct
methoxylation of halide in preformed IMPY derivatives.
[0026] FIG. 7. Synthesis of IMPY Derivatives II using catalytic
substitution of halides by thiolate-based palladium catalysts,
EXAMPLES
[0027] Materials and Methods
[0028] Preparation of Compounds
[0029] Chemicals and Reagents, Common reagents used in the
syntheses were purchased from Aldrich Chemical Company, Fluka
Chemical Company (Milwaukee, Wis.), Acros (Hampton, N.H.), or Strem
Chemicals (Newburyport, Mass.), and were used without further
purification, unless otherwise indicated. DiPPF
[1;1'-bis(diisopropylphosphino)ferrocene) and hexamethylditin were
from Strem Chemicals. 2-Mercaptoethanol, 5-iodopyridin-2-amine,
5-bromopyridin-2-amine, pyrazin-2-amine, 2-bromoacetyl bromide,
o-toluidine, 1-(4-aminophenyl)ethanone,
N-methyl-N-(trimethylstannyl)methanamine, sodium methoxide, and
Pd.sub.2dba.sub.3 (dipalladium dibenzylideneacetone), were from
Aldrich, 2-mercaptoacetamide and 6-aminopyridine-3-carbonitrile
(P4) were from Acros. Water was purified through a Millipore water
purification system, comprising a combination of two filters, one
Rio.TM., one reservoir, and one Milli-Q.RTM., synthesis system
(Bedford, Mass.). Common solvents were obtained from Fisher
Scientific (Pittsburgh, Pa.), Brain tissue from deceased
Alzheimer's Disease (AD) patients was obtained from Brain Bank of
the Clinical Brain Disorders Branch, National Institute of Mental
Health, National Institutes of Health.
[0030] 1-(4-(methylamino)phenyl)ethanone (2),.sup.2
1-(4-amino-3-bromophenyl)ethanone (3), N,2-dimethylbenzenamine
(K2), 2,2,2-trifluoro-N-o-totylacetamide (A3), N-o-tolylacetamide
(A4), 6-aminopyridine-2-carbonitrile (P3),
6-amino-5-bromopyridine-3-carbonitrile (P11),
5-bromo-3-iodopyridin-2-amine (B3), were synthesized as reported in
the literature.
[0031] Instrument and general conditions. Analytical HPLC was
performed using a reverse phase column (X-Tera C18; 5 .mu.m;
10.0.times.250 mm; Waters) eluted with c. ammonia (0.25%) in
acetonitrile-water at 6.2 mL/min. The chromatography system was
fitted with a continuous wavelength UV-vis detector (Beckman System
Gold 168 Detector) and an autosampler (Beckman System Gold 508
Autosampler), For semi-preparative Beckman HPLC, a reverse phase
column (X-Tera C18; 5 .mu.m 19.times.250 mm; Waters) was eluted at
20.0 mL/min. The HPLC system was fitted with a manual injector (5
mL injection loop) and a third delivery pump using acetonitrile as
eluant at 1 mL/min. The purity of compounds was determined with
HPLC monitored for UV absorbance at 280 nm (for IMPY derivatives)
or 254 nm (for other aromatic compounds) and expressed as area
percentage of all peaks. The .sup.1H and .sup.13C NMR spectra of
all compounds were acquired on a Jeol GSX 270 (270 MHz .sup.1H and
68 MHz .sup.13C), on a Bruker DRX 300 (300 MHz .sup.1H and 75 MHz
.sup.13C), on a Bruker DRX 400 (400 MHz .sup.1H and 100 MHz
.sup.13C) and on a Bruker AM500 (500 MHz .sup.1H and 125 MHz
.sup.13C), using the chemical shifts of residual deuterated solvent
as the internal standard; chemical shift (.delta.) data for the
proton and carbon resonance were reported in parts per million
(ppm) relative to the internal standard. Thin-layer chromatography
(TLC) was performed using Silica Gel 60 F254 plates from EM Science
and compounds visualized under UV light at either 250 or 360 nm.
Flash chromatography was carried out using a Biotage Horizon.TM.
HPFC.TM. system (Charlottesville, Va., column sizes: 12
mm.times.150 mm, 25 mm.times.150 mm, 40 Mm.times.150 mm) with
hexanes and ethyl acetate (EtOAc) as eluents with chromatographic
solvent proportions expressed on a volume: volume basis. IR spectra
were recorded using a Perkin-Elmer Spectrum One FT-IR spectrometer,
and UV/vis spectra were recorded using a Lambda 40 UV/vis
spectrometer. Mass spectra were acquired using either Thermo
Finnigan LCQ.sup.DECA LC-MS (MS-HPLC column: Luna C18; 5
.mu.m2.0.times.150 mm; Phenomenex, flow rate; 150 .mu.L/min,
eluent; MeOH and H.sub.2O mixture) or Thermo Finnigan PolarisQ
GC-MS (GC column: capillary RTX-5 ms 30 m.times.0.25 mm, flow rate:
1 mL/min, carrier gas; He), or VG Micromass 7070E and AutoSpec-Q
spectrometers. High-resolution mass spectra (HRMS) were acquired
from Mass Spectrometry Laboratory, University of Illinois at
Urbana-Champaign (Urbana, Ill.). Elemental analyses of selected
compounds were carried out by Midwest Microlab (Indianapolis, Ind.)
or Galbraith Laboratories, Inc, (Knoxville, Tenn.), or Mr Stephen
Boyer, SACS, University of London. The melting points were measured
using Electrothermal Mel-Temp Manual Melting Point Apparatus
(Fisher Scientific), and uncorrected. A CEM Discover microwave
system was used for microwave synthesis (Matthews, N.C.).
[0032] The following syntheses are illustrated by molecular
formulas and reaction schemes in the Figures attached hereto, and
the structures represented by the acronyms and other symbols used
below for the various intermediates and compounds are shown in the
Figures.
[0033] 1-(3-bromo-4-(methylamino)phenyl)ethanone (K3), The compound
was synthesized by bromination of the aryl amine by NBS
(N-Bromosuccinimide).
[0034] The following are syntheses of ketones and
.alpha.-bromoketones according to FIG. 1 as intermediates in the
preparation of the IMPY derivatives.
[0035] N-(4-acetyl-2-methylphenyl)-2,2,2-trifluoroacetamide (K4),
2,2,2-trifluoro-N-o-tolylacetamide (8.0 g; 39.4 mmol) was dissolved
in 20 mL CS.sub.2. Bromoacetic bromide (8.4 g; 68.3 mmol) was added
dropwise. AlCl.sub.3 (16.0 g; 120 mmol) was added in portions, with
drying tube attached to the reaction flask. The reaction mixture
was refluxed overnight. After reaction, the solvent was removed.
Ice and water were added, and some yellow solid Precipitated. The
solid was isolated by filtration, and purified by silica gel column
chromatography, to afford 3.8 g (yield 39%) of product as off-white
solid. .sup.1H NMR (300 MHz; CDCl.sub.3), .delta.8.30 (s, 1H,
Ar--H), 7.91 (brs, 1H, NH), 7.81 (d, .sup.3J.sub.HH=8.2 Hz, 1H,
Ar--H), 7.36 (d, .sup.3J.sub.HH=7.9 Hz, 1H, Ar--H), 2.60 (s, 3H,
COCH.sub.3), 2.36 (s, 3H, --CH.sub.3).
[0036] N-(4-acetyl-2-bromophenyl)-2,2,2-trifluoroacetamide (K5),
1-(4-aminophenyl)ethanone (2.50 g; 18.5 mmol) was dissolved in 100
mL toluene. NBS (3.30 g; 18.8 mmol) was added portionwise, After 15
min stirring at RT, the organic phase was washed with water twice,
and dried over Na.sub.2SO.sub.4. The solvent was removed by rotavap
to get oil, which was purified by silica gel chromatography, to
afford 1.7 g of a white solid, 1-(4-amino-3-bromophenyl)ethanone
(1.7 g; 7.94 mmol), The intermediate was dissolved in
CH.sub.2Cl.sub.2. The reaction mixture was cooled to 0.degree. C..
Trifluoroacetic anhydride (2.0 g; 9.52 mmol) and NEt.sub.3 (1.0 g;
9.90 mmol) were added sequentially. The reaction mixture was
stirred at RT for another hour. The organic phase was washed twice
with water, dried over MgSO.sub.4. The solvent was removed, to
afford 2.4 g (yield 42%) of product as white solid. .sup.1H NMR
(400 MHz, CDCl.sub.3) .delta.8.61 (s, 1H, N-H), 8.46, (d,
.sup.3J.sub.HH=8.6 Hz, 1H, Ar--H), 8.21 (d, .sup.4J.sub.HH2.0 Hz,
1H, Ar--H), 7.95 (dd, .sup.3J.sub.HH=8.6 Hz, .sup.4J.sub.HH=1.9 Hz,
1H, Ar--H), 2.58 (s, 3H, CH.sub.3).
[0037] N-(4-acetyl-2-bromophenyl)-2,2,2-trifluoro-N-methylacetamide
(K6). 1-(3-bromo-4-(methylamino)phenyl)ethanone (1.4 g; 6.14 mmol)
was dissolved in 15 mL CH.sub.2Cl.sub.2. The reaction mixture was
cooled to 0.degree. C. Trifluoroacetic anhydride and Et.sub.3N were
added Sequentially. The mixture mixture was stirred at RT for 3
hrs, washed with water three times, and dried over
Na.sub.2SO.sub.4. The solvent was removed, and product was dried to
afford 1.9 g (yield 96%) of a yellow solid. .sup.1H NMR (300 MHz;
CDCl.sub.3), .delta.8.13 (s, 1H, Ar--H), 7.83 (d,
.sup.3J.sub.HH=8.8 Hz, 1H, Ar--H), 7.02 (d, .sup.3J.sub.HH=8.6 Hz,
1H, Ar--H), 2.91 (s, 3H, NCH.sub.3), 2.53 (s, 3H, COCH.sub.3).
[0038]
N-(4-(2-bromoacetyl)-2-methylphenyl)-2,2,2-trifluoroacetamide (B2).
2,2,2-trifluoro-N-o-tolylacetamide (5.0 g; 24.6 mmol) was dissolved
in 20 mL CS.sub.2. The mixture was cooled below 10.degree. C.
Bromoacetic bromide (10.1 g; 50.0 mmol) was added dropwise.
AlCl.sub.3 (10.0 g; 75.0 mmol) was added in portions, with drying
tube attached to the reaction Flask. The reaction mixture was
refluxed overnight. After the reaction is complete as monitored by
TLC, the upper CS.sub.2 layer was removed. The lower layer was
added into 7.0 mL ice water mixed with HCl. The product was
extracted into CH.sub.2Cl.sub.2, and dried over MgSO.sub.4. The
solvent was removed, and the solid was dried to afford 5.0 g of
crude product, which was purified by silica gel column
chromatography, to afford 3.0 g (yield 38%) of product as yellow
solid. .sup.1H NMR (300 MHz; CDCl.sub.3), .delta.8.40 (s, 1H,
Ar--H), 7.87 (brs+d, .sup.3J.sub.HH=8.0 Hz, 2H, Ar--H), 7.43 (d,
.sup.3J.sub.HH=8.0 Hz, 1H, Ar--H), 4.46 (s, 2H, COCH.sub.2Br), 2.41
(s, 3H, CH.sub.3).
[0039] N-(4-(2-bromoacetyl)-2-methylphenyl)-N-methylacetamide (B3),
.sup.1H NMR (300 MHz; CDCl.sub.3), .delta.7.89 (dd,
.sup.3J.sub.HH=8.0 Hz, .sup.4J.sub.HH=1.7 Hz, 1H, Ar--H), 7.78 (d,
.sup.4J.sub.HH=1.7 Hz, .sup.1H, Ar--H), 7.45 (d, .sup.3J.sub.HH=8.0
Hz, 1H, Ar--H), 4.41 (AB, .sup.2J.sub.HH=11.7 Hz, 2H,
COCH.sub.2Br), 3.20 (s, 3H, NCH.sub.3), 2.33 (s, 3H, Ar-CH.sub.3),
1.78 (s, 3H, COCH.sub.3).
[0040] N-(2-bromo-4-(2-bromoacetyl)phenyl)-2,2,2-trlfluoroacetamide
(B4), N-(4-acetyl-2-bromophenyl)-2,2,2-trifluoroacetamide (12.1 g;
39.0 mmol) and tetrabutylammonium tribromide (18.8 g; 39.0 mmol)
were dissolved separately in ethanol. After combination, and
refluxing for 4 hrs, there was precipitate formation. The solvent
was Removed. NaHCO.sub.3 solution was added to neutralize any
residue acid. The solid was filtered, and washed with
CH.sub.2Cl.sub.2, to afford 7.0 g (yield 46%) of product as pale
green solid. .sup.1H NMR (400 MHz, CDCl.sub.3) .delta.8.64 (s, 1H,
N-H), 8.51 (d, 3J.sub.HH=8.6 Hz, 1H, Ar--H), 8.25 (d,
.sup.4J.sub.HH=2.0 Hz, 1H, Ar--H), 7.98 (dd, .sup.3J.sub.HH=8.6 Hz,
.sup.4J.sub.HH=2.0 Hz, 1H, Ar--H), 4.38 (s, 2H,
--CH.sub.2--Br).
[0041]
N-(2-bromo-4-(2-bromoacetyl)phenyl)-2,2,2-trifluoro-N-methylacetami-
de(B5). CuBr.sub.2 (6.0 g; 26.9 mmol) was suspended in 20 mL ethyl
acetate. The mixture was heated to reflux.
N-(4-acetyl-2-bromophenyl)-2,2,2-trifluoro-N-methylacetamide (4.5
g; 13.9 mmol) dissolved in 20 mL CHCl.sub.3 was added. The mixture
was refluxed for 4 hrs, and filtered. The solvent was removed from
the filtrate to get an orange solid, which was purified by silica
gel column chromatography using the mixture of ethyl acetate and
petroleum ether from 1:100 to 1:50, to afford 1.9 g (yield 34%) of
product as yellow solid. .sup.1H NMR (400 MHz, CDCl.sub.3) .delta.
Isomer .alpha. (80%) 8.33 (d, .sup.4J.sub.HH=2.0 Hz, 1H, Ar--H),
8.02 (dd, .sup.3J.sub.HH=8.2 Hz, .sup.4J.sub.HH=2.0 Hz, 1H, Ar--H),
7.48 (dd, .sup.3J.sub.HH=8.2 Hz, .sup.5J.sub.HH=0.5 Hz, 1H, Ar--H),
4.46 (AB, .sup.2J.sub.HH=11.8 Hz, 1H, CH.sub.2), 4.42 (AB,
.sup.2J.sub.HH=11.8 Hz, 1H, CH.sub.2), 3.36 (d, .sup.4J.sub.HF=0.4
Hz, 3H, CH.sub.3). Isomer .beta. (20%) 8.33 (d, .sup.4J.sub.HH=2.0
Hz, 1H, Ar--H), 8.05 (dd, .sup.3J.sub.HH=8.2 Hz, .sup.4J.sub.HH=2.0
Hz, 1H, Ar--H), 7.44 (dd, .sup.3J.sub.HH=8.2 Hz, .sup.5J.sub.HH=0.5
Hz, 1H, Ar--H), 4.46 (AB, .sup.2J.sub.HH=11.8 Hz, 1H, CH.sub.2),
4.42 (AB, .sup.2J.sub.HH=11.8 Hz, 1H, CH.sub.2), 3.49 (q,
.sup.4J.sub.HF=1.6 Hz,3H, CH.sub.3).
[0042] The following are syntheses of 2-aminopyridine derivatives
according to FIGS. 2, 3, and 4 as further intermediates in the
preparation of the IMPY derivatives and the IMPY derivatives
themselves.
[0043] 5-methoxypyridin-2-amine (P5). Method a:
2-Amino-4-bromopyridine (0.10 g; 0.58 mmol), sodium methoxide (0.16
g; 2.9 mmol) and copper powder nanosized activated (0.11 g; 1.74
mmol) were introduced in a screw cap vial (Pyrex glass) together
with 2.0 mL of anhydrous MeOH and a stirrer bar. The vial was
closed and put in an oil bath at 135.degree. C. and stirred for 14
h. The mixture was cooled, diluted with MeOH (5.0 mL) and filtered
through an SPE silica gel cartridge and the product eluted with
AcOEt. The fractions were collected and evaporated obtaining a
crude of 92 mg of product which was further purified by FCC
(DCM/AcOEt=1:1) to give 26 mg (yield 36%) of the title compound as
brown oil. Method b: 2-Amino-4-bromopyridine (0.10 g; 0.58 mmol),
sodium methoxide (0.16 g; 2.9 mmol) and copper powder nanosized
activated (0.11 g; 1.74 mmol) were introduced in a microwave glass
tube with 1.5 mL of anhydrous DMF and sealed. The tube was
introduced in the microwave cavity and heated for 30 min at
140.degree. C. (140C30M75W300Psi). Although DMF is a high boiling
solvent, high pressure was observed, probably caused by the partial
methanolysis of the DMF resulting in low boiling products such as
methyl formate and dimethylamine. The mixture was diluted with 10
mL of 2 M NH.sub.4Cl solution and extracted 3 times with AcOEt. The
organic phase was washed 2 times with 2 M NH.sub.4Cl solution and 1
time with water to remove the remaining DMF, dried on NaSO.sub.4,
and filtered. After the solvent was removed, the crude product was
purified by FCC (AcOEt) to afford 12 mg (yield 17%) of the title
compound as brown oil. .sup.1H NMR (270 MHz; CDCl.sub.3),
.delta.7.74 (1 H, d, .sup.3J.sub.HH=3.0 Hz), 7.06 (1H, dd,
.sup.3J.sub.HH=9.0 Hz, .sup.4J.sub.HH=3.0 Hz), 6.45 (1 H, d,
.sup.3J.sub.HH=9.0 Hz), 3.95 (2H, bs, NH.sub.2), 3.74 (3H, s,
OCH.sub.3); m/z (EI/MS): 124 (M.sup.+), 109
([M-CH.sub.3].sup.+).
[0044] 5-(ethylthio)pyridin-2-amine (P9). 2-Amino-5-iodo-pyridine
(2.20 g; 10 mmol), sodium ethanethiolate 80% (1.7 g; 16 mmol) and
copper powder (190 mg; 3.00 mmol) were loaded into a 100 mL round
bottom flask under nitrogen. Ethylene glycol (40 mL; 0.25 mol) was
added and the solution stirred at 150.degree. C. for 26 h. The
cooled solution was filtered and partitioned between ethyl acetate
and water twice. The organic layer was dried over barium oxide,
filtered and the solvent removed in vacuo to afford 1.2 g (yield
76%) of product as yellow oil. .sup.1H-NMR (300 MHz; CDCl.sub.3),
.delta.8.06 (1H, d, .sup.4J.sub.HH=2.1 Hz, Ar--H), 7.48 (1H, dd,
.sup.3J.sub.HH=8.7 Hz, .sup.4J.sub.HH=2.4 Hz, Ar--H), 6.42 (1H, dd,
.sup.3J.sub.HH=8.7 Hz, .sup.5J.sub.HH=0.6 Hz, Ar--H), 2.68 (2H, q,
.sup.3J.sub.HH=7.3 Hz, CH.sub.2), 1.15 (3H, t, .sup.3J.sub.HH=7.2
Hz, CH.sub.3).
[0045] 6-amino-3-bromopyridine-2-carbonitrile (P10). .sup.1H NMR
(300 MHz; DMSO-d.sub.6), .delta.7.75 (d, .sup.3J.sub.HH=9.1 Hz, 1H,
Ar--H), 6.69 (d, .sup.3J.sub.HH=9.1 Hz, 1H, Ar--H).
[0046] 5-chloropyrazin-2-amine (P13). Method a: In a double necked
dry flask equipped with condenser and dropping funnel, under
nitrogen flow, were added 2-amino-pyrazine (0.50 g, 5.25 mmol) and
13 mL of anhydrous CHCl.sub.3 previously passed on basic
Al.sub.2O.sub.3. This solution was heated at 60.degree. C. under
stirring. A solution of NCS (0.35 g, 2.12 mmol) in 7 mL of the
anhydrous CHCl.sub.3 was added to the mixture through dropping
funnel during 1.5 hr. After another 30 minutes, the reaction was
stopped and the solvent evaporated. The residue was dissolved in
methanol and absorbed on silica (2 g); This crude product was
purified by FCC (Hexane/DCM/AcOEt=1:1:1) to afford 73 mg (yield
26%) of product as yellow solid. Method b: 2-Amino-pyrazine (0.10
g, 1.05 mmol) and NCS (80 mg, 0.60 mmol) were added in a microwave
tube with 1.5 mL of the anhydrous CHCl.sub.3 and sealed. The tube
was placed in the microwave cavity and heated at 70.degree. C. for
10 min. (70C10M60W300 Psi). After the work-up, the crude (0.15 g)
was purified by FCC (Hexane/DCM/AcOEt=1:1:1) to afford 35 mg (yield
45%) of the product as yellow solid. .sup.1H NMR (270 MHz;
d.sub.6-DMSO), .delta.7.99 (1H, s, H-6), 7.67 (1H, s, H-3), 6.65
(2H, bs, NH.sub.2), m/z (EI-MS): 129 (M.sup.+), 99, 94
([M-Cl].sup.+).
[0047]
4-(6-chloro-H-imidazo[1,2-a]pyridin-2-yl)-N)N-dimethylbenzenamine
(L5). 2-Bromo-1-(4-(dimethylamino)phenylethanone (11) (1.21 g; 5.00
mmol) and 5-chloro-2-aminopyridine (7c) (0.668 g; 5.20 mmol) were
used to give 12c (0.65 g; yield 48%); Mp: 234-236.degree. C.;
.sup.1H NMR (400 MHz, CDCl.sub.3).delta.8.10 (s, 1H, Ar--H), 7.76
(d, .sup.3J.sub.HH=8.9 Hz, 2H, Ar--H), 7.65 (s, 2H, Ar--H), 7.14
(d, .sup.3J.sub.HH=9.2 Hz, 1H, Ar--H), 6.71 (d, .sup.3J.sub.HH=8.6
Hz, 2H, Ar--H), 2.95 (s, 6H, CH.sub.3). .sup.13C.sup.13 NMR (400
MHz, CDCl.sub.3) .delta.149.3, 146.2, 142.6, 125.8 (s, 2C), 124.2,
121.8, 120.0, 118.8, 116.0, 111.2 (s, 2C), 105.7, 39.2 (s, 2C,
NCH.sub.3), m/z (ES-MS): 275.1 (4%), 274.1 (51%), 273.1 (18%),
272.1 (100%, [M+H].sup.+). HRMS m/z (TOF.sup.+): Calc.
C.sub.15H.sub.15N.sub.3Cl=272.0955. Found: 272.0960. Error (ppm):
+1.8.
[0048]
4-(6-fluoro-H-imidazo[1,2-a]pyridin-2-yl)-N,N-dimethylbenzenamine
(L6). 2-Bromo-1-(4-(dimethylamino)-phenylethanone (11) (1.21 g;
5.00 mmol) and 5-fluoro-2-aminopyridine (7d) (0.583 g; 5.17 mmol)
were stirred in reflux ethanol (50 mL) for 4 h. Pale yellow
precipitate formed. NaHCO.sub.3 (0.41 g; 4.9 mmol) was added to the
reaction mixture after cooling (15 min.). The mixture was refluxed
again for another 2 h. After cooling, the solvents were removed.
The solid was washed with water, CH.sub.2Cl.sub.2, and
recrystallized from ethyl acetate, to get 0.42 g of product. Yield
33%. Mp: 228-229.degree. C.; .sup.1H NMR (400 MHz, CD.sub.3OD)
.delta.8.35 (m, .sup.3J.sub.FH=4.2 Hz, .sup.4J.sub.HH=2.4 Hz,
.sup.5J.sub.HH=0.7 Hz, 1H, Ar--H), 7.93 (d, .sup.4J.sub.HH=0.3 Hz,
1H, Ar--H), 7.66-7.62 (m, 2H, Ar--H), 7.41 (m, .sup.3J.sub.HH=9.8
Hz, .sup.4J.sub.FH=5.0 Hz, .sup.5J.sub.HH=0.6 Hz, 1H, Ar--H), 7.14
(m, .sup.3J.sub.HH=9.8 Hz, .sup.3J.sub.FH=8.3 Hz,
.sup.4J.sub.HH=2.4 Hz, 1H, Ar--H), 6.74-6.70 (m, 2H, Ar--H), 2.89
(s, 6H, CH.sub.3). .sup.13C NMR (400 MHz, CDCl.sub.3) .delta.153.2
(d, .sup.1J.sub.FC=236.4 Hz, 1C, Ar), 150.0 (s, 1C, Ar), 147.5 (s,
1C, Ar), 143.1 (s, 1C, Ar), 127.0 (s, 2C, Ar), 121.2 (s, 1C, Ar),
117.2 (d, .sup.3J.sub.FC=9.3 Hz, 1C, Ar), 116.0 (d,
.sup.2J.sub.FC=25.4 Hz,1C, Ar), 112.4 (s, 2C, Ar), 112.0 (d,
.sup.2J.sub.FC=40.6 Hz, 1C, Ar), 107.9 (d, .sup.4J.sub.FC=1.8 Hz,
1C, Ar), 40.5 (s, 2C, CH.sub.3). m/z (ES-MS): 257.1 (7%), 256.1
(100%, [M+H].sup.+). HRMS m/z (TOF.sup.+): Calc.
C.sub.15H.sub.15N.sub.3F=256.1250. Found: 256.1261. Error (ppm):
+4.3.
[0049]
N,N-dimethyl-4-(6-nitro-H-imidazo[1,2-a]pyridin-2-yl)benzenamine
(L7). 2-Bromo-1-(4-(dimethylamino)phenylethanone (11) (0.40 g; 1.65
mmol) and 5-nitro-pyridin-2-amine (7f) (0.30 g; 2.15 mmol) were
stirred in reflux anhydrous acetonitrile (25 mL) at 90-95.degree.
C. ander dinitrogen for 2 h, NaHCO.sub.3 (0.25 g; 2.97 mmol) was
added to the reaction mixture after cooling (15 min). The mixture
was refluxed for another 9 h. After cooling, the mixture was
filtered through a Busch funnel. The precipitate was washed with
acetonitrile and water, to afford 0.11 g of the red product. Yield
23%. Mp: 275-277.degree. C.; .sup.1H NMR (400 MHz, DMSO-d.sub.6)
.delta.9.77 (dd, .sup.4J.sub.HH=2.3 Hz, .sup.5J.sub.HH=0.8 Hz, 1H,
Ar--H), 8.42 (s, 1H, Ar--H), 7.91 (dd, .sup.3J.sub.HH=9.8 Hz,
.sup.4J.sub.HH=2.3 Hz, 1H, Ar--H), 7.83-7.80 (m, 2H, Ar--H),
7.68-7.65 (m, 1H, Ar--H), 6.83-6.80 (m, 2H, Ar--H), 2.97 (s, 6H,
CH.sub.3). .sup.13C.sup.13 NMR (400 MHz, DMSO-d.sub.6)
.delta.150.6, 148.5, 144.9, 136.0, 127.3, 126.9 (s, 2C, Ar), 120.2,
118.4, 115.2, 112.2 (s, 2C, Ar), 109.3, 40.2 (s, 2C, CH.sub.3), m/z
(ES-MS): 284.1 (29%), 283.1 (100%, [M+H].sup.+), HRMS m/z
(TOF.sup.+): Calc. C.sub.15H.sub.15N.sub.4O.sub.2=283.1195. Found
283.1206. Error (ppm); +3.9.
[0050]
2-(4-(dimethylamino)phenyl)-H-imidazo[1,2-a]pyridine-6-carbonitrile
(L8). The same method as described above was used.
2-Bromo-1-(4-(dimethylamino)phenylethanone (11) (1.21 g; 5.00 mmol)
and 6-aminopyridine-3-carbonitrile (7e) (0.63 g; 5.3 mmol) were
used to give 12e (0.85 g; yield 65%); Mp: 269-271.degree. C.;
.sup.1H NMR (400 MHz, CDCl.sub.3) .delta.8.42 (dd,
.sup.4J.sub.HH=1.6 Hz, .sup.5J.sub.HH0.6 Hz, 1H, Ar--H), 7.76-7.73
(m, 2H, Ar--H), 7.72 (s, 1H, Ar--H), 7.58-7.55 (m, 1H, Ar--H), 7.15
(dd, .sup.3J.sub.HH=9.3 Hz, .sup.4J.sub.HH=1.7 Hz, 1H, Ar--H),
6.72-6.69 (m, 2H, Ar--H), 2.95 (s, 6H, CH.sub.3). .sup.13C.sup.13
NMR (400 MHz, CDCl.sub.3) .delta.149.9, 148.1, 143.7, 129.9, 126.3
(s, 2C, Ar), 123.0, 119.2, 116.6, 115.8, 111.2 (s, 2C, Ar), 106.2,
96.9, 39.3 (s, 2C, CH.sub.3). m/z (ES-MS): 364.0 (8%), 264.1 (29%),
263.1 (100%, [M+H].sup.+). HRMS m/z (TOF.sup.+); Calc.
C.sub.16H.sub.15N.sub.4=263.1297. Found: 263.1303.Error (ppm):
+2.3.
[0051]
4-(6-methoxy-H-imidazo[1,2-a]pyridin-2-yl)-N,N-dlmethylbenzenamine
(L9). In a 2-necked round bottomed flask equipped with a condenser
and under nitrogen flow were introduced the
2-amino-5-methoxypyridine (0.15 g, 1.17 mmol), the
2-bromo-4'-dimethylamino-acetophenone (0.31 g, 1.29 mmol) and 14 mL
of absolute EtOH. The reaction mixture was stirred at reflux for 2
h. After it cooled down, NaHCO.sub.3 (0.15 g, 1.75 mmol) was added,
the mixture was refluxed for another 6 hrs. After the solvent was
removed, the residue was dissolved in AcOEt. The organic phase was
washed with water, dried over MgSO.sub.4, and filtered. The solvent
was removed to afford 0.271 g crude product, which was purified by
FCC (DCM/AcOEt=1:1) to give 0.129 g (yield 41%) of the product as
yellow solid, M.p.=177-179.degree. C. .sup.1H NMR (270 MHz;
CDCl.sub.3), .delta.7.82 (d, .sup.3J.sub.HH=8.7 Hz, 2H, Ar--H),
7.71 (s, 1H, H-3), 7.65 (s, 1H, Ar--H), 7.53 (d, .sup.3J.sub.HH=9.6
Hz, 1H, Ar--H), 6.96 (dd, .sup.3J.sub.HH=9.6, .sup.4J.sub.HH=1.8,
1H, Ar--H), 6.81 (d, .sup.3J.sub.HH=8.8, 2H, Ar--H), 3.84 (s, 3H,
OCH.sub.3), 3.02 (s, 6H, NCH.sub.3); .sup.13C{.sup.1H} NMR (125
MHz, CDCl.sub.3), .delta.151.0, 150.4, 149.2, 146.0, 143.0, 126.8
(Ph-2,6), 119.2 (C7), 117.0 (C8), 112.5 (Ph-3,5), 107.6 (C3), 106.2
(C5), 56.2 (OCH.sub.3), 40.5 (NCH.sub.3), m/z(EI-MS): 267
(M.sup.+), 252 (M.sup.+-CH.sub.3); Calc. C.sub.16H.sub.17N.sub.3O;
C, 71.89; H, 6.41; N, 15.72. Found: C, 71.60; H, 6.62; N,
15.96.
[0052] 2-(4-(dimethylamino)phenyl)-H-imidazo[1,2-a]pyridin-6-ol
(L12). In a dried 10 mL flask under nitrogen flow, was added the
6-methoxy-imidazopyridine (62 mg, 0.22 mmol) and 2 mL of anhydrous
DCM. The stirring solution was cooled at -78.degree. C. with a
CO.sub.2/acetone bath and BBr.sub.3 (80 mg, 0.32 mmol) was added
dropwise by a disposable syringe. (Cautionl BBr.sub.3 reacts
violently with water). The reaction mixture was allowed to stir at
RT for 2 hrs, and poured into 15 mL ice/water. The mixture of
CH.sub.2Cl.sub.2 and CH.sub.3OH (15:1) was added and the mixture
stirred for 10 min. The organic phase was separated, dried over
MgSO.sub.4, and filtered. The solvent was removed to give 39 mg
(yield 70%) of the product as yellow solid. M.p.=262-264.degree.
C., .sup.1H NMR (270 MHz; DMSO-d.sub.6), .delta.9.70 (s, 1H, OH),
8.16 (s, 1H, H-3), 8.01 (s, 1H, H-5), 7.71 (d, .sup.3J.sub.HH=8.9
Hz, 2H, Ar--H), 7.45 (d, .sup.3J.sub.HH=9.6 Hz, 1H, Ar--H); 7.03
(d, .sup.3J.sub.HH=9.4, 1H, Ar--H), 6.78 (d, .sup.3J.sub.HH=8.9 Hz,
2H, Ar--H), 2.95 (s, 6H, NCH.sub.3); .sup.13C{.sup.1H} NMR (125
MHz, DMSO-d.sub.6), .delta.150.2, 146.2, 143.2, 140.3, 126.4
(Ph-2,6), 120.7 (C7), 120.4, 115.3 (C8), 112.4 (Ph-3,5), 110.5
(C3), 107.8 (C5), 39.9 (NCH.sub.3). m/z (CI-MS): 254 ([M+1].sup.+),
238 ([M-CH.sub.3].sup.+).
[0053]
4-(6-(ethylthio)-H-imidazo[1,2-a]pyridin-2-yl)-N,N-dimethylbenzenam-
ine (T7). 2-Bromo-1-(4-(dimethylamino)phenylethanone (11) (1.21 g;
5.00 mmol) and 5-(ethylthio)pyridin-2-amine (7h) (0.80 g; 5.2 mmol)
were used to give 12h (0.91 g; yield 61%); mp: 168-171.degree. C.;
.sup.1H NMR (400 MHz, DMSO-d.sub.6) .delta.8.56 (dd,
.sup.4J.sub.HH=1.9 Hz, .sup.5J.sub.HH=0.9 Hz, 1H, Ar--H), 8.14 (s,
1H, Ar--H), 7.78-7.74 (m, 2H, Ar--H), 7.51-7.49 (m, 1H, Ar--H), 723
(dd, .sup.3J.sub.HH=9.3 Hz, .sup.4J.sub.HH=1.9 Hz, 1H, Ar--H), 6.76
(m, 2H, Ar--H), 2.94 (s, 6H, CH.sub.3), 2.92 (q, .sup.3J.sub.HH=7.3
Hz, 2H, CH.sub.2), 1.21 (t, .sup.3J.sub.HH=7.3 Hz, 3H, CH.sub.3).
.sup.13C NMR (100 MHz, DMSO-d.sub.6) .delta.150.1, 145.7, 143.6,
128.0, 127.3, 126.5 (s, 2C, Ar), 121.5, 118.3, 116.1, 112.2 (s, 2C,
Ar), 107.1, 40.0 (s, 2C, CH.sub.3), 28.4 (s, 1C, CH.sub.2), 14.4
(s, 1C, CH.sub.3). m/z (ES-MS): 300.1 (6%), 299.1 (39%), 298.1
(100%, [M+H].sup.+). HRMS m/z (TOF.sup.+): Calc.
C.sub.17H.sub.20N.sub.3S=298.1378. Found: 298.1386. Error (ppm):
+2.7.
[0054] 4-(6-bromo-8-iodoindolizin-2-yl)-N,N-dimethylbenzenamine
(L13). 2-Bromo-1-(4-(dimethylamino)phenylethanone (11) (1.21 g;
5.00 mmol) and 2-amino-3-iodo-5-bromopyridine (7i) (1.58 g; 5.3
mmol) were used to give 12i (1.21 g; yield 55 %); mp:
218-220.degree. C.; .sup.1H NMR (400 MHz, CDCl.sub.3) .delta.8.17
(d, .sup.4J.sub.HH=1.7 Hz, 1H, Ar--H), 7.85-7.81 (m, 2H, Ar--H),
7.79 (s, 1H, Ar--H), 7.66 (d, .sup.4J.sub.HH=1.7 Hz, 1H, Ar--H),
6.79 (d, .sup.3J.sub.HH=7.9 Hz, 2H, Ar--H), 2.99 (s, 3H, CH.sub.3),
.sup.13C NMR (100 MHz, CDCl.sub.3) .delta.150.5, 147.9, 143.9,
135.8, 127.5 (s, 2C, Ar), 125.4, 121.5, 112.8 (s, 2C, Ar), 108.8,
106.0, 84.0 (s, 1C, Ar-I), 40.9 (s, 1C, CH.sub.3). m/z (ES-MS):
444.9 (6%), 443.9 (99%), 442.9 (9%), 441.9 (100%, [M+H].sup.+),
HRMS m/z (TOF.sup.+): Calc. C.sub.15H.sub.14N.sub.3BrI=441.9416.
Found: 441.9398. Error (ppm): -4.1.
[0055] 6-bromo-2-(4-(dimethylamino)phenyl)indolizine-8-carbonitrile
(L14). 2-Bromo-1-(4-(dimethylamino)phenylethanone (11) (1.21 g;
5.00 mmol) and2-amino-5-bromonicotinonitrile (7j) (1.03 g; 5.2
mmol) were used to give 12j (0.77 g; yield 45%); mp:
240-246.degree. C.; .sup.1H NMR (400 MHz, DMSO-d.sub.6) .delta.9.12
(d, .sup.4J.sub.HH=1.8 Hz, 1H, Ar--H), 8.32 (s, 1H, Ar--H), 8.13
(d, .sup.4J.sub.HH=1.8 Hz, 1H, Ar--H), 7.81 (d, .sup.3J.sub.HH=8.9
Hz, 2H, Ar--H), 6.79 (d, .sup.3J.sub.HH=8.9 Hz, 2H, Ar--H), 2.96
(s, 3H, CH.sub.3), .sup.13C NMR (100 MHz, DMSO-d.sub.6)
.delta.150.6, 147.4, 141.3, 133.6, 131.1, 127.0 (s, 2C, Ar), 119.9,
114.8, 112.1 (s, 2C, Ar), 109.0, 103.5, 99.9, 39.9 (s, 2C,
NCH.sub.3). m/z (ES-MS): 443.9 (4%), 441.9 (3%), 344.0 (11%), 343.0
(98%), 342.0 (11%), 341.0 (100%, [M+H].sup.+), HRMS m/z
(TOF.sup.+): Calc. C.sub.16H.sub.14N.sub.4Br=341.0402.Found:
341.0389. Error (ppm): -3.8.
[0056]
N-(2-bromo-4-(6-bromo-H-imidazo[1,2-a]pyridin-2-yl)phenyl)-2,2,2-tr-
ifluoro-N-methylacetamide (L15).
N-(2-Bromo-4-(2'-bromoacetyl)phenyl)-2,2,2-trifluoro-N-methylacetamide
(4a) (4.0 g; 9.93 mmol) and 5-bromopyridin-2-amine (7b) (1.7 g;
9.83 mmol) were dissolved in MeOH (20 mL). The reaction mixture was
refluxed for 8 h and solvent then removed. A small amount of
CH.sub.2Cl.sub.2 was added and the resultant precipitate filtered
off to afford 13a (2.8 g; yield 59%) as a yellow solid; mp:
95-98.degree. C.; .sup.1H NMR (400 MHz, CD.sub.3OD) .delta. Isomer
.alpha. (84%) 8.73-8.72 (m, 1H, Ar--H), 8.34-8.31 (m, 2H, Ar--H),
8.01 (dd, .sup.3J.sub.HH=8.2 Hz, .sup.4J.sub.HH=2.0 Hz, 1H, Ar--H),
7.58 (d, .sup.3J.sub.HH=8.3 Hz, 1H, Ar--H), 7.55-7.52 (m, 1H,
Ar--H), 7.45 (dd, .sup.3J.sub.HH=9.6 Hz, .sup.4J.sub.HH=1.9 Hz, 1H,
Ar--H), 2.66 (s, 3H, CH.sub.3). Isomer .beta. (16%) 8.73-8.72 (m,
1H, Ar--H), 8.34-8.31 (m, 2H, Ar--H), 8.03 (dd,
.sup.3J.sub.HH=8.0Hz, .sup.4J.sub.HH=1.9 Hz, 1H, Ar--H), 7.55-7.52
(m, 1H, Ar--H), 7.51 (d, .sup.3J.sub.HH=8.3 Hz, 1H, Ar--H), 7.44
(dd, .sup.3J.sub.HH=9.6 Hz, .sup.4J.sub.HH=1.9 Hz, 1H, Ar--H), 2.66
(s, 3H, CH.sub.3). .sup.13C NMR (100 MHz, DMSO-d.sub.6) .delta.
Isomer .alpha. 155.5 (q, .sup.2J.sub.CF=35.1 Hz, 1C, CO), 143.5,
142.3, 137.9, 136.3, 130.8, 129.8, 128.6, 125.8, 122.8, 117.9,
115.9 (q, .sup.1J.sub.CF=286 Hz, 1C, CF.sub.3), 111.1, 106.5, 37.7
(s, 1C, CH.sub.3). Isomer .beta. 155.5 (q, .sup.2J.sub.CF=35.1 Hz,
1C, CO), 143.4, 142.6, 139.8, 135.6, 129.9, 129.4, 128.4, 127.1,
126.3, 121.3, 117.8, 115.9 (q, .sup.1J.sub.CF=286 Hz, 1C,
CF.sub.3), 110.8, 109.4, 37.7 (s, 1C, CH.sub.3). m/z (ES-MS): 479.9
(27%), 477.9 (100%, [M+H].sup.+), 475.9 (28%), HRMS m/z
(TOF.sup.+): Calc.
C.sub.16H.sub.11N.sub.3OF.sub.3Br.sub.2=475.9221. Found: 475.9228.
Error (ppm): +1.5.
[0057]
N-(4-(6-bromoindolizin-2-yl)-2-methylphenyl)-N-methylacetamide
(L16). N-(4-(2'-Bromoacetyl)-2-methylphenyl)-N-methylacetamide (4c)
(1.0 g; 3.52 mmol) and 5-bromopyridin-2-amine (7b) (0.73 g; 4.22
mmol) were used. Yield 37%, M.p. 197-198.degree. C.; .sup.1H NMR
(400 MHz, DMSO-d.sub.6) .delta. Isomer .alpha. (90%) 8.88 (dd,
.sup.4J.sub.HH=2.0 Hz, .sup.5J.sub.HH=0.8 Hz, 1H, Ar--H), 8.39 (d,
.sup.5J.sub.HH=0.4 Hz, 1H, Ar--H), 7.89 (dd, .sup.3J.sub.HH=7.8 Hz,
.sup.4J.sub.HH=1.8 Hz, 1H. Ar--H), 7.84 (d, .sup.4J.sub.HH=1.8 Hz,
1H. Ar--H), 7.58-7.54 (m, 1H, Ar--H), 7.43 (d, .sup.3J.sub.HH=8.0
Hz, 1H, Ar--H), 7.37 (dd, .sup.3J.sub.HH=9.5 Hz, .sup.4J.sub.HH=2.0
Hz, 1H, Ar--H), 3.11 (s, 3H, NCH.sub.3), 2.21 (s, 3H, Ar-CH.sub.3),
1.70 (s, 3H, CH.sub.3). Isomer .beta. (10%) 8.87 (dd,
.sup.4J.sub.HH=1.9 Hz, .sup.5J.sub.HH=0.8 Hz, 1H, Ar--H), 8.32 (d,
.sup.5J.sub.HH=0.3 Hz, 1H, Ar--H), 7.76 (dd, .sup.3J.sub.HH=7.8 Hz,
.sup.4J.sub.HH=1.8 Hz, 1H. Ar--H), 7.73 (d, .sup.4J.sub.HH=1.8 Hz,
1H. Ar--H), 7.56-7.53 (m, 1H, Ar--H), 7.36 (dd, .sup.3J.sub.HH=9.5
Hz, .sup.4J.sub.HH=2.0 Hz, 1H, Ar--H), 7.31 (d, .sup.3J.sub.HH=8.0
Hz, 1H, Ar--H), 3.28 (s, 3H, NCH.sub.3), 2.13 (s, 3H, Ar-CH.sub.3),
1.70 (s, 3H, CH.sub.3), .sup.13C NMR (100 MHz, DMSO-d.sub.6)
.delta. Isomer .alpha. 169.0 (s, 1C, CO), 144.2, 143.3, 143.2,
134.7, 133.0, 131.7, 127.9, 126.9, 125.3, 125.1, 117.6, 109.8,
106.0, 35.3 (s, 1C, NCH.sub.3), 21.6 (s, 1C, CH.sub.3), 16.7 (s,
1C, Ar-CH.sub.3). Isomer .beta. 169.6 (s, 1C, CO), 144.6, 143.8,
143.2, 134.8, 132.2, 130.9, 127.8, 126.8, 124.5, 124.1, 117.5,
109.3, 105.9, 38.7 (s, 1C, NCH.sub.3), 21.9 (s, 1C, CH.sub.3), 17.1
(s, 1C, Ar-CH.sub.3), m/z (ES-MS); 361.0 (4%), 360.0 (100%,
[M+H].sup.+), 359.0 (47%), 358.0 (96%). HRMS m/z (TOF.sup.+); Calc.
C.sub.17H.sub.17N.sub.3OBr=358.0555. Found: 358.0557. Error (ppm):
+0.6.
[0058]
N-(4-(6-bromoindolizin2-yl)-2-methylphenyl)-2,2,2-trifluoroacetamid-
e (L17).
N-(4-(2-bromoacetyl)-2-methylphenyl)-2,2,2-trifluoroacetamide (3.0
g; 9.26 mmol) and 5-bromopyridin-2-amine (1.5 g; 8.67 mmol) were
dissolved in 20 mL ethanol. The mixture was refluxed overnight, and
the solvent was removed. CH.sub.2Cl.sub.2 was added to precipitate
a solid, which was filtered and washed with CH.sub.2Cl.sub.2, to
afford 1.0 g (yield 28%) of product as a yellow powder. .sup.1H NMR
(300 MHz; DMSO-d.sub.6), .delta.11.1 (s, 1H, NH), 9.14 (s, 1H,
Ar--H), 8.60 (s, 1H, Ar--H), 7.90-7.76 (m, 4H, Ar--H), 7.46 (d,
.sup.3J.sub.HH=8.0 Hz, 1H, Ar--H), 2.19 (s, 3H, Ar-CH.sub.3).
[0059]
2-bromo-4-(6-bromo-H-imidazo[1,2-a]pyridin-2-yl)-N-methylbenzenamin-
e (L18).
N-(2-bromo-4-(6-bromo-H-imidazo[1,2-a]pyridin-2-yl)phenyl)-2,2,2--
trifluoro-N-methylacetamide (1.0 g; 2.10 mmol) and K.sub.2CO.sub.3
(2.0 g; 14.5 mmol) were suspended in 30 mL ethanol and 15 mL water.
The mixture was refluxed over 8 hrs. The organic solvent was
removed, and CH.sub.2Cl.sub.2 was added. The organic phase was
washed with H.sub.2O twice, and dried over MgSO.sub.4. The solvent
was removed to afford 0.60 g (yield 80%) of product as a yellow
solid. .sup.1H NMR (300 MHz; DMSO-d.sub.6), .delta.8.80 (s, 1H,
Ar--H), 8.20 (s, 1H, Ar--H), 7.98 (d, .sup.4J.sub.HH=1.8 Hz, 1H,
Ar--H), 7.76 (dd, .sup.3J.sub.HH=8.5 Hz, .sup.4J.sub.HH=1.8 Hz, 1H,
Ar--H), 7.49 (d, .sup.3J.sub.HH=9.5 Hz, 1H, Ar--H), 7.30 (dd,
.sup.3J.sub.HH=9.4 Hz, .sup.4J.sub.HH=1.7 Hz, 1H, Ar--H), 6.67 (d,
.sup.3J.sub.HH=8.5 Hz, 1H, Ar--H), 2.78 (s, 3H, CH.sub.3).
[0060]
4-(6-bromo-H-imidazo[1,2-a]pyridin-2-yl)-N,2-dimethylbenzenamine
(L19).
N-(4-(6-Bromoimidazo[2-a]pyridin-2-yl)-2-methylphenyl)-N-methylace-
tamide (13c, 1.0 g; 2.79 mmol) and KOH (1.0 g; 17.8 mmol) were
used, to afford 0.68 g. Yield 77%, M.p. 165-166.degree. C.; .sup.1H
NMR (400 MHz, DMSO-.sub.6) .delta.8.82 (dd, .sup.4J.sub.HH=1.8 Hz,
.sup.5J.sub.HH=0.7 Hz, 1H, Ar--H), 8.27 (s, 1H, Ar--H), 7.56 (d,
.sup.3J.sub.HH=9.5 Hz, 1H, Ar--H), 7.33 (dd, .sup.3J.sub.HH=9.5 Hz,
.sup.4J.sub.HH=1.9 Hz, 1H, Ar--H), 7.10-7.09 (m, 2H, Ar--H), 7.01
(dd, .sup.3J.sub.HH=8.3 Hz, .sup.5J.sub.HH=0.32 Hz, 1H, Ar--H),
5.12 (s, 1H, N--H), 2.82 (s, 1H, CH.sub.3), 2.10 (s, 1H, CH.sub.3),
.sup.13C NMR (100 MHz, DMSO-d.sub.6) .delta.147.8, 146.4, 143.1,
131.9, 129.8, 127.3, 126.6, 121.8, 117.5, 113.2, 108.9, 105.7,
105.6, 30.2 (s, 1C, N--CH.sub.3), 17.5 (s, 1C, Ar-CH.sub.3), m/z
(ES-MS): 319.0 (17%), 318.0 (96%), 317.0 (22%), 316.0 (100%,
[M+H].sup.+). HRMS m/z (TOF.sup.+): Calc.
C.sub.15H.sub.15N.sub.3Br=316.0449. Found: 316.0453. Error (ppm):
+1.3.
[0061] 4-(6-bromo-H-imidazo[1,2-a]pyridin-2-yl)-2-methylbenzenamine
(L20).
N-(4-(6-bromo-H-imidazo[1,2-a]pyridin-2-yl)-2-methylphenyl)-2,2,2-trifluo-
roacetamide (1.0 g; 2.51 mmol) and K.sub.2CO.sub.3 (4.0 g; 29.0
mmol) were suspended in 30 mL ethanol and 15 mL water. The mixture
was refluxed overnight. The solvent was removed, and
CH.sub.2Cl.sub.2 was added. The organic phase Was washed with
H.sub.2O twice, and dried over MgSO.sub.4. The solvent was removed
to afford a solid which was purified by silica gel column
chromatography to obtain 0.60 g (yield 79%) of product as yellow
solid. .sup.1H NMR (300 MHz; DMSO-d.sub.6), .delta.8.81 (s, 1H,
Ar--H), 8.13 (s, 1H, Ar--H), 7.49 (d, .sup.3J.sub.HH=9.5 Hz, 1H,
Ar--H), 7.30 (d, .sup.3J.sub.HH=9.5 Hz, 1H, Ar--H), 7.25 (s, 1H,
Ar--H), 7.03 (d, .sup.3J.sub.HH=7.4 Hz, 1H, Ar--H), 6.95 (d,
.sup.3J.sub.HH=7.7 Hz, 1H, Ar--H), 2.06 (s, 3H, Ar-CH.sub.3).
[0062]
4-(6-chloroimidazo[1,2-a]pyrazin-2-yl)-N,N-dimethylbenzenamine
(L21). In a double necked round bottomed flask equipped with
condenser and under nitrogen flow were introduced
5-chloro-2-amino-pyrazine (61 mg, 0.47 mmol),
2-bromo-4'-dimethylamino-acetophenone (170 mg, 0.71 mmol) and
anhydrous acetonitrile (7 mL). The reaction mixture was refluxed
for 3 hrs. After it cooled down, NaHCO.sub.3 (71 mg, 0.85 mmol) was
added. The reaction was refluxed for another 6 hrs. The solvent was
removed and the crude product (221 mg) was purified by FCC
(hex/DCM/AcOEt=2:2:1), to afford 9 mg (yield 7%) of the product as
yellow solid. .sup.1H NMR (270 MHz; CDCl.sub.3), .delta.8.82 (s,
1H, Ar--H), 8.09 (s, 1H, Ar--H), 7.822 (s, 1H, Ar--H), 7.818 (d,
.sup.3J.sub.HH=8.9 Hz, 2H, Ar--H), 6.77 (d, .sup.3J.sub.HH=8.7 Hz,
2H, Ar--H), 3.02 (s, 6H, NCH.sub.3). m/z (CI-MS): 273
([M+1].sup.+), 261,239 ([M--Cl+1].sup.+).
[0063]
4-(6-chloroimidazo[1,2-a]pyrimidin-2-yl)-N,N-dimethylbenzenamine
(L22). 2-Amino-5-chloro-pyrimidine (0.150 g, 1.16 mmol),
2-bromo-4'-dimethylamino-acetophenone (0.56 g, 2.32 mmol) and 16 mL
of anhydrous acetonitrile were added to a double-neck flask
equipped with a condenser under dinitrogen. A small amount of
anhydrous DMF (0.5 mL) was added to completely dissolve the amine.
After 3 hrs of stirring and refluxing, the mixture was cooled down
and NaHCO.sub.3 (0.19 g, 2.32 mmol) was added. The reaction mixture
was refluxed for another 6 hrs and was filtered hot on a Busch
funnel to collect the precipitate, which was washed with portions
of EtOH, water, EtOH, and dried overnight, to afford 72 mg (yield
23%) of the product as yellow powder. M.p.=256-258.degree. C.
.sup.1H NMR (270 MHz; CDCl.sub.3), .delta.8.37 (d, 2H,
.sup.4J.sub.HH=2.7 Hz, Ar--H), 7.88 (d, .sup.3J.sub.HH=9.2 Hz, 2H,
Ar--H), 7.67 (s, 1H, H-3), 6.79 (d, .sup.3J.sub.HH=8.7, 2H, Ar--H),
3.01 (s, 6H, NCH.sub.3). .sup.13C{.sup.1H} NMR(125 MHz .sup.13C,
CDCl.sub.3), .delta.150.5, 149.3, 147.9 (C7), 147.0, 129.6 (C5),
127.5 (Ph-2,6), 117.5, 112.8 (Ph-3,5), 106.2 (C--Cl), 104.8 (C3),
40.7 (NCH.sub.3), m/z (CI-MS): 273 ([M+1].sup.+), 239
([M--Cl+1].sup.+). Calc. C.sub.14H.sub.13ClN.sub.4; C, 61.75; H,
4.82; N, 20.59. Found: C, 58.32; H, 4.88; N, 18.81.
[0064] General procedure for thiolate substitution: A 10 mL
microwave tube was charged with Pd.sub.2dba.sub.3 (13-58 mg,
0.014-0.063 mmol, 10-20 mol % Pd), DiPPF (6-27 mg, 0.014-0.063
mmol, 10-20 mol %), IMPY derivative (0.14--0.32 mmol), and
tin-thiolate (0.29-0.63 mmol). The tube was capped, and put in a
microwave system for desired temperature and time as specified in
the text. A small sample of the resulting suspension was analyzed
by HPLC to confirm the conversion. The suspension was partitioned
between CHCl.sub.3 and K.sub.2CO.sub.3 solution. The organic layer
was dried over MgSO.sub.4 and filtered. The solvent was removed,
and the residue was dissolved in DMSO, and loaded on a reversed
phase HPLC. After proper band was collected, the solvents were
removed, and the product was dried by azetrope with CH.sub.3CN to
provide the desired product.
[0065]
2-(2-(4(methylamino)pheny)-H-imidazo[1,2-a]pyridin-6-ylthio)acetami-
de (T1).
4-(6-Iodo-H-imidazo[1,2-a]pyridin-2-yl)-N-methylbenzenamine (100
mg, 0.29 mmol), N-methyl-N-(trimethylstannyl)methanamine (60 mg,
0.29 mmol), 2-mercaptoacetamide (26 mg, 0.29 mmol),
Pd.sub.2(dba).sub.3 (26.2 mg, 0.029 mmol), DiPPF (12 mg, 0.029
mmol), and 8.0 mL of toluene were used.
[0066] Mp: 198-202.degree. C.; .sup.1H NMR (400 MHz, DMSO-d.sub.6)
.delta.8.68 (d, .sup.4J.sub.HH=4.4 Hz, 1H, Ar--H), 8.11 (s, 1H,
Ar--H), 7.68 (d, .sup.3J.sub.HH=8.0 Hz, 2H, Ar--H), 7.48 (d,
.sup.3J.sub.HH=9.6 Hz, 1H, Ar--H), 7.26 (d, .sup.3J.sub.HH=9.2 Hz,
1H, Ar--H), 7.13 (s, 1H, Ar--H), 6.59 (d, .sup.3J.sub.HH=8.0 Hz,
2H, Ar--H), 5.86 (s, 2H, NH.sub.2), 3.53 (s, 2H, CH.sub.2S), 2.71
(s, 3H, CH.sub.3). .sup.13C{.sup.1H} NMR (400 MHz, DMSO-d.sub.6)
.delta.169.8 (--CO--N), 149.8, 146.2, 143.6, 128.0, 127.7, 126.6,
120.9, 118.2, 115.9, 111.6, 106.8, 38.5 (C--S), 29.6 (NCH.sub.3).
m/z (LC-MS): 314.3 (21%), 313.2 (100%, [M+H].sup.+), 255.5 (32%,
[M--CH.sub.2CONH.sub.2].sup.+). HRMS m/z (TOF.sup.+): Calc.
C.sub.16H.sub.17N.sub.4OS=313.1123. Found: 313.1126. Error (ppm):
+0.9.
[0067]
2-(2-(4-(dimethylamino)phenyl)-H-imidazo[1,2-a]pyridin-6-ylthio)ace-
tamide (T2).
4-(6-Iodo-H-imidazo[1,2-a]pyridin-2-yl)-N,N-dimethylbenzenamine (50
mg, 0.14 mmol), N-methyl-N-(trimethylstannyl)methanamine (30 mg,
0.14 mmol), 2-mercaptoacetamide (13 mg, 0.14 mmol),
Pd.sub.2(dba).sub.3 (13 mg, 0.014 mmol), DiPPF (6 mg, 0.014 mmol),
and 5.0 mL of toluene were used.
[0068] Mp: 180-183.degree. C.; .sup.1H NMR (400 MHz, DMSO-d.sub.6)
.delta.8.60 (s, 1H, Ar--H), 8.17 (s, 1H, Ar--H), 7,77 (d,
.sup.3J.sub.HH=8.8 Hz, 2H, Ar--H), 7.50 (d, .sup.3J.sub.HH=7.9 Hz,
1H, Ar--H), 7.27 (d, .sup.3J.sub.HH=9.4 Hz, 1H, Ar--H), 7.13 (brs,
2H, NH.sub.2), 6.78 (d, .sup.3J.sub.HH=8.7 Hz, 2H, Ar--H), 3.54 (s,
2H, CH.sub.2S), 2.94 (s, 6H, N(CH.sub.3).sub.2), .sup.13C{.sup.1H}
NMR (400 MHz, DMSO-d.sub.6) .delta.168.7 (--CO--N), 149.0, 144.7,
142.5, 129.4, 127.8, 126.9, 126.6, 125.3, 120.3, 117.2, 114.9,
111.1, 109.5, 106.1, 38.5 (C--S), 37.3 (NCH.sub.3). m/z (LC-MS);
329.3 (7%), 328.2 (18%), 327.2 (100%, [M+H].sup.+), 325.1 (12%),
323.4 (7%), 321.7 (7%), 320.9 (5%), 269.4 (14%,
[M--CH.sub.2CONH.sub.2].sup.+). HRMS m/z (TOF.sup.+); Calc.
C17H.sub.19N.sub.4OS=327.1280. Found: 327.1278. Error (ppm);
-0.5.
[0069]
2-(2-(4-(methylamino)phenyl)-H-imidazo[1,2-a]pyridin-6-ylthio)ethan-
ol (T3).
4-(6-Iodo-H-imidazo[1,2-a]pyridin-2-yl)-N-methylbenzenamine (100
mg, 0.29 mmol),N-methyl-N-(trimethylstannyl)methanamine (60 mg,
0.29 mmol), 2-mercaptoethanol (23 mg, 0.29 mmol),
Pd.sub.2(dba).sub.3 (28 mg, 0.031 mmol), DiPPF (22 mg, 0.039 mmol),
and 5.0 mL of toluene were used.
[0070] Mp: 158-161.degree. C.; .sup.1H NMR (400 MHz, CD.sub.3OD)
.delta.8.51 (s, 1H, Ar--H), 7.95 (s, 1H, Ar--H), 7.68 (d,
.sup.3J.sub.HH=8.6 Hz, 2H, Ar--H), 7.45 (d, .sup.3J.sub.HH=8.6 Hz,
1H, Ar--H), 7.34 (d, .sup.3J.sub.HH=8.6 Hz, 1H, Ar--H), 6.67 (d,
.sup.3J.sub.HH=8.6 Hz, 2H, Ar--H), 3.69 (t, .sup.3J.sub.HH=6.5 Hz,
2H, OCH.sub.2), 3.01 (t, .sup.3J.sub.HH=6.5 Hz, 2H, SCH.sub.2),
2.81 (s, 3H, NCH.sub.3), .sup.13C{.sup.1H} NMR (400 MHz,
CD.sub.3OD) .delta.151.8, 148.0, 145.7, 130.9, 129.5, 128.1, 122.6,
121.2, 116.5, 113.4, 108.5, 61.5 (C-O), 38.8 (C-S), 30.6
(NCH.sub.3). m/z (LC-MS): 302.2 (5%), 301.3 (15%), 300.2 (100%,
[M+H].sup.+). HRMS m/z (TOF.sup.+): Calc.
C.sub.16H.sub.18N.sub.3OS=300.1171. Found: 300.1163. Error (ppm):
-2.5.
[0071]
2-(2-(4-(dimethylamino)phenyl)-H-imidazo[1,2-a]pyridin-6-ylthio)eth-
anol(T4).
4-(6-Iodo-H-imidazop[1,2-a]pyridin-2-yl)-N,N-dimethylbenzenamine
(50 mg, 0.14 mmol), N-methyl-N-(trimethylstannyl)methanamine (29
mg, 0.14 mmol), 2-mercaptoethanol (11 mg, 0.14 mmol),
Pd.sub.2(dba).sub.3 (13 mg, 0.014 mmol), DiPPF (6 mg, 0.014 mmol),
and 5.0 mL of toluene were used.
[0072] Mp: 183-185.degree. C.; .sup.1H NMR (400 MHz, CD.sub.3OD)
.delta.8.50 (s, 1H, Ar--H), 8.05 (s, 1H, Ar--H), 7.78 (d,
.sup.3J.sub.HH=8.9 Hz, 2H, Ar--H), 7.45 (d, .sup.3J.sub.HH=8.9 Hz,
1H, Ar--H), 7.35 (d, .sup.3J.sub.HH=8.9 Hz, 1H, Ar--H), 6.83 (d,
.sup.3J.sub.HH=8.9 Hz, 2H, Ar--H), 3.70 (t, .sup.3J.sub.HH=6.7 Hz,
2H, OCH.sub.2), 3.03 (t, .sup.3J.sub.HH=6.7 Hz, 2H, SCH.sub.2),
3.00 (s, 6H, NCH.sub.3), .sup.13C{.sup.1H} NMR (400 MHz,
DMSO-d.sub.6) .delta.150.1, 145.7, 143.6, 128.1, 127.4, 126.4,
121.5, 118.5, 116.1, 112.2, 107.1, 59.8 (C--O), 55.9 (C--S), 37.2
(NCH.sub.3). m/z (LC-MS): 316.1 (5%), 315.2 (23%), 314.2 (100%,
[M+H].sup.+). HRMS m/z (TOF.sup.+): Calc.
C.sub.17H.sub.20N.sub.3OS=314.1327. Found: 314.1317. Error (ppm):
-3,4.
[0073]
4-(6-(4-methoxybenzylthio)-H-imidazo[1,2-a]pyridin-2-yl)-N,N-dimeth-
ylbenzenamine (T5).
4-(6-Iodo-H-imidazo[1,2-a]pyridin-2-yl)-N,N-dimethylbenzenamine (80
mg, 0.22 mmol), N-methyl-N-(trimethylstannyl)methanamine (46 mg,
0.22 mmol), (4-methoxyphenyl)methanethiol (34 mg, 0.22 mmol),
Pd.sub.2(dba).sub.3 (40 mg, 0.044 mmol), DiPPF (18.4 mg, 0.044
mmol), and 5.0 mL of toluene were used.
[0074] Mp: 184-186.degree. C.; .sup.1H NMR(400 MHz, CD.sub.3OD)
.delta.8.43 (s, 1H, Ar--H), 8.10 (s, 1H, Ar--H), 7.75 (d,
.sup.3J.sub.HH=8.4 Hz, 2H, Ar--H), 7.47 (d, .sup.3J.sub.HH=8.0 Hz,
1H, Ar--H), 7.19 (d, .sup.3J.sub.HH=8.0 Hz, 1H, Ar--H), 7.16 (d,
.sup.3J.sub.HH=8.0 Hz, 2H, Ar--H), 6.83 (d, .sup.3J.sub.HH=8.0 Hz,
1H, Ar--H), 6.77 (d, .sup.3J.sub.HH=8.0 Hz, 1H, Ar--H), 4.13 (s,
2H, CH.sub.2S), 3.71 (s, 3H, CH.sub.3O), 2.94 (s, 6H, NCH.sub.3).
.sup.13C{.sup.1H} NMR (400 MHz, DMSO-d.sub.6) .delta.158.2, 150.0,
145.6, 143.5, 129.9, 129.2, 128.2, 127.7, 126.3, 121.3, 118.0,
115.8, 113.9, 112.1, 107.0, 54.8 (OCH.sub.3), 39.9 (NCH.sub.3),
38.3 (C--S). m/z (LC-MS): 392.1 (7%), 391.1 (23%), 390.1 (100%,
[M+H].sup.+), HRMS m/z (TOF.sup.+): Calc.
C.sub.23H.sub.24N.sub.3OS=390.1640. Found: 390.1634. Error (ppm);
-0.6.
[0075]
N,N-dimethyl-4-(6-(methylthio)-H-imidazo[1,2-a]pyridin-2-yl)benzena-
mine (T6).
4-(6-Iodo-H-imidazo[1,2-a]pyridin-2-yl)-N,N-dimethylbenzenamine (90
mg, 0.25 mmol), 1,2-dimethyldisulfane (28 mg, 0.30 mmol),
1,1,1,2,2,2-hexamethyldistannane (97 mg, 0.30 mmol),
Pd.sub.2(dba).sub.3 (22.8 mg, 0.025 mmol), DiPPF (10.5 mg, 0.025
mmol), and 4.0 mL of toluene were used.
[0076] Mp: 155-164.degree. C.; .sup.1H NMR (400 MHz, CD.sub.3OD)
.delta.8.34 (s, 1H, Ar--H), 7.96 (s, 1H, Ar--H), 7.73 (d,
.sup.3J.sub.HH=9.4 Hz, 2H, Ar--H), 7.44 (d, .sup.3J.sub.HH=9.4 Hz,
1H, Ar--H), 7.26 (dd, .sup.3J.sub.HH=9.4 Hz, .sup.4J.sub.HH=2.2 Hz,
1H, Ar--H), 6.82 (d, .sup.3J.sub.HH=8.9 Hz, 2H, Ar--H),
.sup.13C{.sup.1H} NMR (400 MHz, CD.sub.3OD) .delta.152.3, 147.6,
145.6, 129.0, 128.1, 125.9, 124.6, 122.8, 116.7, 113.9, 108.8, 40.9
(C-S), 17.9 (NCH.sub.3), m/z (LC-MS): 286.2 (4%), 285.3 (13%),
284.2 (100%, [M+H].sup.+). HRMS m/z (TOF.sup.+): Calc.
C.sub.16H.sub.18N.sub.3S=284.1221. Found: 284.1215. Error (ppm):
-2.4.
[0077]
2-(2-(3-methyl-4-(methylamino)phenyl)-H-imidazo[1,2-a]pyridin-6-ylt-
hio)acetamide (T8).
4-(6-Bromo-H-imidazo[1,2-a]pyridin-2-yl)-N2-dimethylbenzenamine
(100 mg, 0.32 mmol), N-methyl-N-(trimethylstannyl)methanamine (99
mg, 0.48 mmol), 2-mercaptoacetamide (43 mg, 0.48 mmol),
Pd.sub.2(dba).sub.3 (43 mg, 0.048 mmol), DiPPF (20 mg, 0.048 mmol),
and 6.0 mL of toluene were used.
[0078] Mp: 181-182.degree. C.; .sup.1H NMR (400 MHz, CD.sub.3OD)
.delta.8.58 (s, 1H, Ar--H), 8.10 (s, 1H, Ar--H), 7.49 (d,
.sup.3J.sub.HH=8.0 Hz, 1H, Ar--H), 7.41 (d, .sup.3J.sub.HH=8.0 Hz,
1H, Ar--H), 7.17 (s, 1H, Ar--H), 7.12 (d, .sup.3J.sub.HH=8.0 Hz,
1H, Ar--H), 7.06 (d, .sup.3J.sub.HH=8.0 Hz, 1H, Ar--H), 3.46 (s,
1H, NH), 3.55 (s,2H, SCH.sub.2), 2.93 (s, 3H, NCH.sub.3), 2.16 (s,
3H, Ar-CH.sub.3). .sup.13C{.sup.1H} NMR (400 MHz, DMSO-d.sub.6)
.delta.168.7 (--CO--), 146.6, 144.9, 142.4, 131.0, 128.6, 127.1,
126.6, 120.5, 117.5, 115.3, 112.0, 107.5, 104.5, 37.2 (CH.sub.2S),
29.0 (NCH.sub.3), 16.3 (CH.sub.3). m/z (LC-MS): 329.2 (6%), 328.3
(22%), 327.2 (100%, [M+H].sup.+). HRMS m/z (TOF.sup.+): Calc.
C.sub.17H.sub.19N.sub.4OS=327.1280. Found: 327.1278. Error (ppm):
-0.4.
[0079]
2-(2-(3-methyl-4-(methylamino)phenyl)-H-imidazo[1,2-a]pyridin-6-ylt-
hio)ethanol (T9).
4-(6-Bromo-H-imidazo[1,2-a]pyridin-2-yl)-N,2-dimethylbenzenamine
(100 mg, 0.32 mmol), N-methyl-N-(trimethylstannyl)methanamine (132
mg, 0.63 mmol), 2-mercaptoethanol (50 mg, 0.63 mmol),
Pd.sub.2(dba).sub.3 (58 mg, 0.063 mmol), DiPPF (27 mg, 0.063 mmol),
and 6.0 mL of toluene were used.
[0080] Mp: 149-153.degree. C.; .sup.1H NMR (400 MHz, CD.sub.3OD)
.delta.8.55 (s, 1H, Ar--H), 8.10 (s, 1H, Ar--H), 7.50 (d,
.sup.3J.sub.HH=8.0 Hz, 1H, Ar--H), 7.38 (d, .sup.3J.sub.HH=8.0 Hz,
1H, Ar--H), 7.18 (s, 1H, Ar--H), 7.15 (d, .sup.3J.sub.HH=8. 0 Hz,
1H, Ar--H), 7.08 (d, .sup.3J.sub.HH=8.0 Hz, 1H, Ar--H), 3.80 (t,
.sup.3J.sub.HH=7.2 Hz, 2H, CH.sub.2O), 3.03 (t, .sup.3J.sub.HH=7.2
Hz, 2H, CH.sub.2S), 2.94 (s, 3H, NCH.sub.3), 2.16 (s, 3H,
Ar-CH.sub.3). .sup.13C{.sup.1H} NMR (400 MHz, DMSO-d.sub.6)
.delta.147.7, 146.0, 143.4, 132.2, 129.7, 128.2, 127.4, 121.6,
118.8, 116.5, 113.1, 108.6, 105.6, 59.5 (OCH.sub.2), 37.1
(CH.sub.2S), 30.1 (NCH.sub.3), 17.5 (CH.sub.3), m/z (LC-MS): 329.2
(6%), 328.3 (22%), 327.2 (100%, [M+H].sup.+), m/z (LC-MS): 316.2
(8%), 315.2 (24%), 314.2 (100%, [M+H].sup.+), HRMS m/z (TOF.sup.+);
Calc. C.sub.17H.sub.20N.sub.3OS=314.1327. Found: 314.1316. Error
(ppm): -3.5.
[0081] Isolation of human AD amyloid. Postmortem brain times were
obtained from a confirmed AD patient at the age of 80. Pre- and
postmortem consent was obtained from next of kin, and brains were
removed at autopsy, sectioned and frozen at -70.degree. C. Bags of
brain tissue were removed from the freezer to a -20.degree. C.
freezer one day before, and then placed on ice 3 h before the
experiment. While working on ice, the brain tissue was cut into
small sections, and meningies from outer layer of cortex and from
fissures between the lobes of the gray matter was removed. After
removing the white matter, the grey matter were chopped into small
pieces and placed into plastic bags. The tissue was stored in
-70.degree. C. freezer for further use.
[0082] In a typical isolation, 60 g of brain tissue were used.
Initially, an equal volume of 1% SDS was added to homogenize. More
1% SDS was added later until 200 mL. The mixture was vortexed,
distributed evenly among 10 Beckman ultracentrifuge tubes, and
centrifuged by a Coulter type 55.2 Ti ultracentrifuge rotor using
RPM: 40,000; G force; 180,000; temp: 4.degree. C.; time: 30 min.
The collected bottom pellets were further homogenized and
centrifuged twice more as described above using 50 mL and 20 mL of
1% SDS each, and finally in 20 mL 1.2 M sucrose. The supernatant
along the top pellet was discarded. The pellets at the bottom were
homogenized and centrifuged in 20 mL 1.9 M sucrose. The top pellets
were collected using a cell scrapper, and the bottom pellets were
discarded. The collected top pellets were homogenized and
centrifuged using 20 mL deionized water. The bottom pellets were
further homogenized and centrifuged using 1.3 M sucrose solution
into one centrifuge tube. The bottom pellets were homogenized and
centrifuged using 20 mL deionized water. The collected pellets were
homogenized in 30 mL of 50 mM Tris-HCl, pH 8.0, 2 mM CaCl.sub.2.
Collagenase CLS3 (120 mg) and DNAase I (10-15 mg) were added. The
mixture was vortexed and incubated for 24-56 hrs at 37.degree. C.
in a water bath with shaking. After incubation, the suspension was
filtered through 41 .mu.m Milllpore filter paper. The collected
filtrate was centrifuged and the bottom pellets were homogenized
and centrifuged one more time using the Tris-HCl solution. The
supernatant was discarded, and the pellets were homogenized and
centrifuged using 10 mL 1.9 M sucrose. The top pellet and the 1.9M
sucrose layer were transferred to a new tube (this is to remove
heavy particles such as metals and salts). Deionized water (40 mL)
was added. After homogenization and centrifugation, the collected
pellets were put into 500 .mu.L of deionized water to form a
homogeneous suspension. It was stored in a refrigerator or freezer
for further use.
[0083] In vitro Binding Assay
[0084] The amyloid suspension prepared above was diluted by a
factor of 40,000 to 50,000 in PBS, and 800 .mu.L suspension was
used in each tube. [.sup.3H]6-OH-BTA1 with a concentration of 1
mCi/mL stock solution was diluted using ethanol to give an
intermediate solution of 1 .mu.Ci/100 .mu.L, which was further
diluted using PBS to result in a dilute stock solution of
2.7.times.10.sup.-2 .mu.Ci/100 .mu.L, and 100 .mu.L was used in
each tube (2 vials with 100 .mu.L of [.sup.3H]6-OH-BTA-1 each and
scintillation fluid as references). Cold 6-OH-BTA-1 or other
displacer was dissolved in ethanol to result in a stock solution of
1.times.10.sup.-3 M, which was further diluted using PBS or ethanol
to result in solutions with concentrations ranging from
1.times.10.sup.-5 to 10.sup.-10 M, and 100 .mu.L was used in each
tube. After assembly, the tubes were vortexed, and incubated for 3
hrs at room temperature. After separation using a cell harvester,
the filter paper (GF/B filter paper pretreated with 0.5% polyimine
solution) was washed with 10% ethanol in PBS (3.times.3 mL). The
filters were placed into 20 mL glass vials and 10 mL scintillation
fluid each was added. After overnight incubation, the samples were
counted. The data were analyzed using GraphPad Prism 4 or
KaleidaGraph 3.6.
[0085] Results and Discussion.
[0086] Chemical Synthesis
[0087] Synthesis of Bromoketones. The nucleus of
imidazo[1,2-a]pyridines is made through the condensation between a
2-amino pyridine and an .alpha.-haloketone in a manner described
before..sup.2 The reaction tolerates a variety of substituents,
including amide and imine groups in both substrates. Two strategies
have been developed to synthesize the .alpha.-bromoketones (FIG.
1). One is from bromination of ketones. Mono-methylation of the
amino group of 1-(4-aminophenyl)ethanone follows the method
established, to give 1-(4-methylaminophenyl)ethanone, KI..sup.2
Ortho-bromination of the aromatic amine was accomplished using NBS,
a mild bromination agent for activated phenyl group, to give K2and
K3..sup.3 The amino group was protected using either
trifluoroacetyl or acetyl group to give K5 and K6 before
bromination of the .alpha.-methyl ketone. Three bromination
procedures have been reported..sup.4 One is through tetra-n-butyl
ammonium tribromide, the second is through copper (II) bromide, and
the third is through the reduction of di-bromides, to afford
.alpha.-bromoketones B2, B3, B4, B5. The other method to synthesize
the .alpha.-bromoketones is through Friedel-Crafts reaction to
introduce .alpha.-bromoacetyl group directly in the aromatic
nucleus. The aromatic amino group in A1 and A2 were protected using
trifluoroacetyl or acetyl group, to give A3 and A4. Friedel-Crafts
reactions using trifluoroacetyl or bromoacetyl bromide catalyzed by
AlCl.sub.3 in CS.sub.2 went smoothly to give B2, B3, B4,
B5..sup.5-7
[0088] Synthesis of 2-Aminopyridines and 2-Aminopyrazines. The
synthetic methods of pyridine derivatives depend on the
substituents on the ring (FIG. 2). For CN group, halide
substitution using CuCN is an optimal choice, such as the synthesis
of P3.sup.8-10 and P4(available from Acros). Some halo group could
be introduced through electrophilic substitution of either
bromination or iodination, such as the synthesis of P10, P11 and
P12.
[0089] Substitution of halide by OCH.sub.3 and SEt group is more
challenging. The first attempts to prepare P5 by a nucleophilic
aromatic displacement of the bromide with a methoxy group catalyzed
by Cu powder in a conventional way.sup.11 or with microwave heating
or an ultrasound bath were frustrated by the apparent unreactivity
of the substrate. An attempt was carried out with an ultrasound
high power probe (Branson 450 Sonifier) beaming the 40 dB sound
waves directly in the solution, but even this technology did not
enable the expected transformation (eq 1).
##STR00003##
[0090] Another literature procedure to perform aromatic
substitutions with the methoxy anion was followed..sup.12 The
method uses salts of copper (I) and a small amount of ester to form
a stabilized tetrahedral adduct which should act as a powerful
methoxide donor (FIG. 3). Unfortunately, due to the presence of the
free amino group on the bromopyridine and the ethyl acetate as
co-catalyst, the reaction product was just the acetamide of the
substrate (eq 2).
##STR00004##
[0091] Attempts to improve the result were made by increasing the
equivalents of catalyst Used. It was believed that copper could be
coordinated and thus inactivated by the nitrogen atoms of the
substrate. By this means the expected product was achieved in 36%
yield (eq 3). The reaction mixture, after being heated for 14 hrs,
still contained a significant amount of starting material together
with black polymers. The reaction was quenched to avoid further
formation of polymers and degradation of the formed product. In
order to decrease the reaction time, the transformation in eq 3 was
carried out with microwave irradiation according to Table 3.
##STR00005##
TABLE-US-00004 TABLE 3 Microwave reaction of eq 3 at 410.degree. C.
Starting Ratio of Time (h) Solvent Recovered
Substitution:Reduction:Polymer 1.0 MeOH Nothing small:small:large
0.5 DME 70% 0:0:large 0.5 DMF Trace 17%:0:large
[0092] Some modifications to this method were tried to increase the
yield. 5-Iodo-2-aminopyridine was used in the hope that the weaker
C-I bond could help to increase the rate of the displacement. The
consumption of aromatic iodide was faster (6 h) but unfortunately
only polymers formed. Decreasing the reaction temperature did not
help (eq 4).
##STR00006##
[0093] A different strategy was designed using 5-pyridyl boronate
and subsequently oxidized to P5 in the presence of NaOCH.sub.3.
After a first attempt to prepare the boronate P6 in DMSO.sup.13
with conventional heating failed, the same coupling was tried with
microwave heating using DME as the solvent..sup.14 This procedure
gave the desired boronate in 20 min instead of 20 h, but only in
10% yield due to problems found in the work-up (eq 5).
##STR00007##
[0094] The free amino group was protected to avoid undesirable
coordination of the Pd catalyst and simplify the reaction work-up.
In addition, 5-iodo-2-aminopyridine was used to facilitate the Pd
insertion in the C-halogen bond. Therefore P1 was protected with
trifluoroacetic anhydride to give trifluoroacetamide P7 in 77%
yield. This was reacted with pinacolborane in dioxane at 80.degree.
C. for 3 h to give the protected pyridyl boronate P8 in 67% yield
(eq 6)..sup.15
##STR00008##
[0095] Oxidation Studies did not generate any desired product. The
route was Discontinued.
[0096] Substitution of halide by SEt group was similar to that of
OCH.sub.3 group when using copper as catalysts. Although syntheses
of P9.sup.16;17 have been reported, the reported methods made use
of copper as catalyst and methanol as solvent at 150.degree. C.
under pressure..sup.18 Due to the inconvenience of the procedure,
other solvents were evaluated to replace the low boiling point
methanol. Ethylene glycol proved to be an excellent choice in this
reaction, giving high yield in a short reaction time. The benefits
of ethylene glycol as solvent in other copper or
copper(I)-catalyzed reaction have been noted..sup.19 Ethylene
glycol was found to be the ligand and solvent when CuI was used as
catalyst, to give the aromatic thio-ether,
2-amino-5-ethylthio-pyridine.
[0097] The synthesis of 5-chloro-2-aminopyrazine described in the
literature is not a very efficient multi-step process..sup.20
Direct chlorination of the 2-aminopyrazine using
N-chlorosuccinimide was used, although the method is known to give
over chlorinated by-products. Initial attempt showed that the
reaction gave a lot of black polymers after a few min., with the
main product being 3,5-dichloro-2-amino pyrazine. Only small amount
of 5-chloro-2-aminopyrazine was isolated (eq 7). This reaction
could not be scaled up because it was not reproducible. To avoid
over chlorination, milder conditions were examined. No reaction was
observed at RT, but a complete transformation into the dichloro
derivative was observed at 50.degree. C.
##STR00009##
[0098] When the distilled chloroform used for the reaction was
pre-treated over basic Al.sub.2O.sub.3, the black polymers formed
in the reaction decreased dramatically. When NCS was added slowly
into the refluxing 2-aminopyrazine, the product was isolated at
26%. Moreover, when microwave irradiation was used at 70.degree. C.
for 10 min., the yield reached 45% (FIG. 4).
[0099] Synthesis of IMPY Derivatives I. Most of the IMPY
derivatives were synthesized through the direct condensation of
.alpha.-bromoketones and 2-amino-pyridines (FIG. 5)..sup.2 In
selected examples, when trifluoroacetyl group was used to protect
aromatic amino group, no base was needed for the condensation. The
difficulty facing the synthesis of L22 came from separation. When
excess amine was used, the product could not be separated from the
starting amine. When excess .alpha.-bromoketone was used, a
persistent amount of the starting pyrimidine existed. A small
amount of DMF was added in the reaction mixture to dissolve the
amine completely, and to induce the precipitation of the product at
the same time. By filtering this solution hot, even before reaction
complete, we were able to isolate the desired imidazopyrimidine L22
in 23% yields (eq 8).
##STR00010##
[0100] Imidazopyrazine L21 was synthesized from condensation of
chloropyrazine P13 and bromoacetophenone B1 (eq 9). The amine 67
was very unreactive though it is very soluble in acetonitrile. Long
reaction time and stronger bases such as triethylamine were
examined with limited success.
##STR00011##
[0101] The synthesis of L12 was through demethylation of the
methoxy group in L9. The most effective reagent to perform the
reaction is BBr.sub.3 in DCM (eq 10)..sup.21
##STR00012##
[0102] The trifluoroacetyl group on the aromatic amino group in
L15, L16, and L17, was easily removed under mild condition of
K.sub.2CO.sub.3 in methanol or ethanol (FIG. 4). This provides an
excellent method for the synthesis of IMPY derivatives with amino
group(s).
[0103] Synthesis of IMPY Derivatives II. For L9, we also attempted
direct methoxylation of halide in pre-formed IMPY derivatives.
Unfortunately the reaction produced a low yield mixture of reduced
(L11), cine (L10) and ipso (L9) substituted analogues which were
difficult to separate (FIG. 6). Catalytic substitution of halides
by thiolate based on palladium-catalysts was developed to overcome
the reduction of the halide (FIG. 7)..sup.22 The new method has
been applied to the synthesis of a number of IMPY derivatives with
thiolate group.
[0104] An advantage of the IMPY derivatives of the present
invention relative to the prior art compound [.sup.11C]PIB
(6-OH-BTA-1) is that the derivatives of the present invention can
be easily labeled with [.sup.18F], which has a half-life of about
two hours as compared with twenty minutes for [.sup.11C]. A process
for radiolabeling the IMPY derivatives with either label is shown
below.
##STR00013##
[0105] Isolation of Human Amyloid
[0106] A method for isolating human .beta.-amyloid plaques from
brain tissue.sup.23;24 was modified by incorporating repeated
homogenization and centrifugation in 1% SDS buffer and by using
DNase and collagenase for digestion. The concentration of
.beta.-amyloid plaques was gauged from the concentration of
A.beta..sub.1-40 and A.beta..sub.1-42 monomers, using ELISA to
detect dissolved plaques in formic acid.
[0107] In Vitro Assay
[0108] Determination of the binding affinities of new ligands for
.beta.-amyloid is the first step in selecting candidate
radioligands for PET studies in humans. Three types of amyloid
plaques have been used to assay ligand binding in vitro, namely
synthetic aggregates of A.beta..sub.1-40, A.beta..sub.1-42, amyloid
plaques from transgenic mice and amyloid plaques from human
Alzheimer's disease (AD) brain tissue. Results from the three types
and also from different batches of synthetic amyloid plaques vary
with respect to binding site concentration..sup.25 These findings
may reflect variations in binding site architecture, though no
significant difference in ligand binding affinity has been detected
among the three types of amyloid plaques for the binding site
typical of 6-OH-BTA-1..sup.26 Since the ultimate test for a
radioligand is successful application in humans, in vitro
evaluation using human AD brain tissue is highly appropriate.
However, it should be noted that other proteins with similar
binding sites might interfere with the binding of the ligand to
.beta.-amyloid plaques. This may then be reflected in a lower
percentage of displaceable radioactivity in the binding
assay..sup.27 The use of isolated human amyloid plaques may also
help to identify other native binding sites.
[0109] Isolated plaques were used in developing an in vitro binding
assay. Tritiated-6-OH-BTA-1 was selected as the reference
radioligand based on its use in human PET imaging and high
affinity..sup.28 Displacement curves were created using
non-radioactive 6-OH-BTA-1 and other novel ligands.
[0110] The displacement of [.sup.3H]6-OH-BTA-1 by non-radioactive
6-OH-BTA-1 or other ligands resulted in classical displacement
curves. Some lipophilic ligands required ethanol in the medium to
increase their solubility and to effect displacement of reference
radioligand. Non-radioactive 6-OH-BTA-1 achieved >95%
displacement of reference radioligand, indicating that the presence
of competing binding sites for the isolated amyloid plaques was
negligible. This assay could therefore be used to screen compounds
for amyloid binding without interference from other proteins, such
as tau-tangles. This displacement curve was also analyzed using a
homologous displacement mathematical model to extract the B.sub.max
of the amyloid plaques. The measured B.sub.max is linear with the
amount of amyloid used in the experiment. The ratio between
B.sub.max and the amount of A.beta..sub.1-42 monomer measured by
ELISA is about 1:2, somewhat less than that reported
previously..sup.25 The denaturing agents used to aid dissolution of
the amyloid plaques may account for the observed difference.
[0111] The binding affinities for a variety of IMPY derivatives are
listed in Table 3. In this table, Ki is a measure (in nM units) of
the binding affinity of the compound toward beta-amyloid plaques in
AD brain tissue, measured through a competitive radioligand
displacement in vitro assay using [.sup.3H]6-OH-BTA-1 as the
reference radioligand. The lipophilicity of the compound is
represented by cLogD.sub.7.4 which is calculated through a software
package (ACD/LogD version 8.0) at pH=7.4, a close mimic of
physiological conditions.
##STR00014##
TABLE-US-00005 TABLE 4 IMPY derivatives. Ligand R.sup.1 R.sup.2
R.sup.3 R.sup.4 R.sup.5 cLogD.sub.7.4 Ki (nM) L1 (IMPY) I H H Me Me
4.37 .+-. 0.88 8.9 .+-. 0.7 L4 Br H H Me Me 4.11 .+-. 0.88 5.9 .+-.
0.4 L5 Cl H H Me Me 3.93 .+-. 0.84 24.2 .+-. 5.6 L6 F H H Me Me
3.38 .+-. 0.88 13.0 .+-. 1.6 L7 NO.sub.2 H H Me Me 3.09 .+-. 1.30
7.6 .+-. 0.7 L8 CN H H Me Me 2.80 .+-. 1.31 8.2 .+-. 1.0 L9 OMe H H
Me Me 3.05 .+-. 1.30 38.5 .+-. 5.0 L12 OH H H Me Me 1.26 .+-. 1.29
177 .+-. 31 T7 SEt H H Me Me 4.24 .+-. 1.31 8.3 .+-. 0.5 L13 Br I H
Me Me 5.17 .+-. 0.93 183 .+-. 61 L14 Br CN H Me Me 3.28 .+-. 1.35
>180 L18 Br H Br H Me 4.30 .+-. 0.94 7.4 .+-. 0.6 L19 Br H Me H
Me 4.16 .+-. 0.88 >1000 L20 Br H Me H H 3.28 .+-. 0.87 658 .+-.
47 T1 SCH.sub.2CONH.sub.2 H H H Me 1.59 .+-. 1.33 1840 .+-. 497 T2
SCH.sub.2CONH.sub.2 H H Me Me 2.02 .+-. 1.33 391 .+-. 76 T3
SCH.sub.2CH.sub.2OH H H H Me 2.54 .+-. 1.35 645 .+-. 75 T4
SCH.sub.2CH.sub.2OH H H Me Me 2.96 .+-. 1.35 88 .+-. 6 T5
SCH.sub.2C.sub.6H.sub.4OCH.sub.3 H H Me Me 4.76 .+-. 1.35 8.3 .+-.
1.8 T6 SMe H H Me Me 3.67 .+-. 1.31 7.9 .+-. 0.6 T8
SCH.sub.2CONH.sub.2 H Me H Me 2.12 .+-. 1.33 >1000 T9
SCH.sub.2CH.sub.2OH H Me H Me 3.07 .+-. 1.35 >1000 L2 I H H Me
CH.sub.2CH.sub.2F 4.52 .+-. 0.92 31 .+-. 5 L3 I H H Me
CH.sub.2CH.sub.2CH.sub.2F 4.90 .+-. 0.92 41 .+-. 5
[0112] The BTA derivatives were synthesized through the
condensation of 2-amino-aryl-thiols and aryl carboxylic acid under
acidic conditions.
##STR00015##
TABLE-US-00006 TABLE 5 Compound oLogD.sub.7.4 Ki (nM) L21 2.54 .+-.
1.30 88 .+-. 22 L22 3.14 .+-. 1.43 147 .+-. 20 P1 4.36 5.7 .+-. 0.7
PIB 3.31 7.23 .+-. 1.0
[0113] Comparison of the Ki values of L1, L18 and L19 in Table 4
shows that the "isosteric effect" works partially. Since Br and Me
are similar in size, the data show that a polar group increases
binding. Similar analysis of the binding affinities of L1, L5, L6,
and T7 shows that the thiol ether group provides optimal electronic
and steric effects for binding, but the effect disappears as soon
as a hydrophilic group is attached (c.f. results for T4 and T8).
Increasing the size of substituents on the aromatic amino group
decreases binding affinity, as reflected in L1, L2 and L3. However,
there is substantial requirement for size in the 6-position (c.f.
L6, L12 and L9). For L12, the combination of small and polar
properties for the 6-substituent abolishes all binding affinity.
Interestingly, introduction of a CN group into 8-position also
completely removes the binding affinity (as in L14),
reflecting-minimal tolerance for any substituents at the periphery
of the IMPY skeleton.
[0114] The reference citations above are identified below under
Reference List No. 1.
Reference List No. 1
[0115] 1. Selkoe, D. J. Alzheimer's disease; genes, proteins, and
therapy, Physiol Rev, 2001, 81, 741-766.
[0116] 2. Cai, L.; Chin, F. T.; Pike, V. W.; Toyama, H.; Liow, J.
S.; Zoghbi, S. S.; Modell, K.; Briard, E.; Shetty, H. U.; Sinclair,
K.; Donohue, S.; Tipre, D.; Kung, M. P.; Dagostin, C.; Widdowson,
D. A.; Green, M.; Gao, W.; Herman, M. M.; Ichise, M.; and Innis, R.
B. Synthesis and evaluation of two .sup.18F-labeled
6-iodo-2-(4'-N,N-dimethylamino)phenylimidazo[1,2-a]pyridine
derivatives as prospective radioligands for beta-amyloid in
Alzheimer's disease. J. Med. Chem. 2004, 47, 2208-2218.
[0117] 3. Hu, Jianguo, Li, Zuoy, Li, Yulin, Tang, Luoxiang, and Wu,
Meijuan. Method of synthesizing
5-[1-hydroxy-2-(isopropylamino)ethyl]aminobenzonitrile. Faming
Zhuanli Shenqing Gongkai Shuomingshu
CN95-103024-19950403[CN1120535A19960417]. 1996. CAN130;139172.
[0118] 4. Cai, L.; Brouwer, C.; Sinclair, K.; Cuevas, J.; and Pike,
V. W. Titanium(IV) chloride-promoted synthesis of new
imidazo[1,2a]pyridine derivatives under microwave conditions.
Synthesis 2005, 61, 0000.
[0119] 5. Hu, Z. Process for synthesizing ketoprofen. Faming
Zhuanli Shenqing Gongkai Shuomingshu
CN-95-109877-19950824[CN1143624A19970226]. 1997. CAN128:153928.
[0120] 6. Kuhla, D. E., Campbell, H. F., Studt, W. L., and Molino,
B. F., Bicyolic heteroaryl thiazole compounds and their cardiotonic
uses. PCT Int.Appl. WO 85-US2522-19851218[WO8603749A1-19860703].
1986. CAN105:226538.
[0121] 7Otsuka-Pharmaceutical-Co, Ltd. Carbostyril derivatives and
a cardiotonic composition containing them. Belg.
BE82-207321-19820215[BE892148A1-19820816]. 1982. CAN98:34510.
[0122] 8. Ellis, G. P. and Romneyalexander, T. M. Cyanation of
aromatic halides. Chem. Rev. 1987, 87, 779-794.
[0123] 9Liang, C. -H., Duffield, J., Romero, A., Chiu, Y. -H.,
Rabuka, D., Yao, S., Sucheck, S., Marby, K, Shue, Y. -K., Ichikawa,
Y., and Hwang, C. -K. Preparation of macrolide erythronolide
carbamates as antitumor and antibacterial agents. (Optimer
Pharmaceuticals, Inc. USA. 2004-US6645[2004080391], 108. 2004, WO,
CAN141;296244, Mar. 5, 2004.
[0124] 10. Ueno, T., Kimura, Y., and Kasuga, Y. Process for the
preparation of Aminocyanopyridines. (Nippon Fine Chemical Co., Ltd.
Japan and Mitsubishi Chemical Corp.). 2000-128683[2001302639], 4.
2001, JP, CAN135:331348, Apr. 28, 2000.
[0125] 11. Van de Poel, H.; Guillaumet, G.; and Viaud-Massuard, M.
C. Synthesis of melatonin analogues derived from furo[2,3-b]- and
[2,3-c]pyridines by use of a palladium-copper catalyst system.
Heterocycles 2002, 57, 55-71.
[0126] 12. Capdevielle, P. and Maumy, M. Esters are effective
cocatalysts in copper-catalyzed methanolysis of aryl bromides.
Tetrahedron Letters 1993, 34, 1007-1010.
[0127] 13. Ishiyama, T.; Murata, M.; and Miyaura, N.
Palladium(O)-catalyzed cross-coupling reaction of alkoxydiboron
with haloarenes--A direct procedure for arylboronic esters. J. Org.
Chem. 1995, 60, 7508-7510.
[0128] 14. Appukkuttan, P.; Van der Eycken, E.; and Dehaen, W.
Microwave enhanced formation of electron rich arylboronates,
Synlett 2003, 1204-1206.
[0129] 15. Baudoin, O.; Guenard, D.; and Gueritte, F.
Palladium-catalyzed borylation of ortho-substituted phenyl halides
and application to the one-pot synthesis of 2,2'-disubstituted
biphenyls. J. Org. Chem. 2000, 65, 9268-9271.
[0130] 16. Sundberg, R. J.; Dahlhausen, D. J.; Manikumar, G.;
Mavunkel, B.; Biswas, A.; Srinivasan, V.; King, F., Jr.; and Waid,
P. Preparation of 2-aryl- and
2-(aryloxymethyl)imidazo[1,2-a]pyridines and related compounds. J.
Heterocycl. Chem. 1988, 25, 129-137.
[0131] 17. Gol'dfarb, Y.; Stoyanovich, F. M.; Marakatkina, M. A.;
and Gorushkina, G. I. Synthesis of
6-(alkylthio)imidazo[1,2-a]pyridines. Khimiya Geterotsiklicheskikh
Soedinenii 1979, 634-638.
[0132] 18. Bochis, R. J.; Olen, L. E.; Waksmunski, F. S.; Mrozik,
H.; Eskola, P.; Kulsa, P.; Wilks, G.; Taylor, J. E.; Egerton, J.
R.; Ostlind, D. A.; and Olson, G. Substituted
Imidazo[1,2-a]pyridine-2-carbamate anthelmintics. J. Med. Chem.
1981, 24, 1518-1521.
[0133] 19. Enguehard, C.; Allouchi, H.; Gueiffier, A.; and
Buchwald, S. L. Easy access to novel substituted
6-aminoimidazo[1,2-a]pyridines using palladium- and
copper-catalyzed animations. J. Org. Chem. 2003, 68, 4367-4370.
[0134] 20. Barlin, G. B.; Davies, L. P.; Ireland, S. J.; Ngu, M. M.
L.; and Zhang, J. K. Imidazo[1,2-b]pyridazines .12. Syntheses and
central-nervous-system activities of some substituted
imidazo[1,2-b]pyridazines and related imidazo[1,2-a]pyridines,
imidazo[1,2-a]pyrimidines and imidazo[1,2-a]pyrazines, Austr. J.
Chem. 1992, 45, 877-888.
[0135] 21. Vickery, E. H.; Pahler, L. F.; and Eisenbraun, E. J.
Selective O-demethylation of catechol ethers--comparison of boron
tribromide and iodotrimethylsilane. J. Org. Chem. 1979, 44,
4444-4446.
[0136] 22. Cai, L.; Cuevas, J.; Peng, Y.; and Pike, V. W. Rapid
palladium-catalyzed cross-coupling in the synthesis of aryl
thioethers under microwave conditions. Org. Letters 2005.
[0137] 23. Soderberg, L.; Zhukareva, V.; Bogdanovic, N.; Hashimoto,
T.; Winblad, B.; Iwatsubo, T.; Lee, V. M.; Trojanowski, J. Q.; and
Naslund, J. Molecular identification of AMY, an Alzheimer disease
amyloid-associated protein. J. Neuropathol. Exp. Neurol. 2003, 62,
1108-1117.
[0138] 24. Roher, A. E. Structural alterations in the peptide
backbone of beta-amyloid core protein may account for its
deposition and stability in Alzheimer's disease. J. Biol. Chem.
1993, 268, 3072-3083.
[0139] 25, Mathis, C. A.; Wang, Y.; Holt, D. P.; Huang, G. F.;
Debnath, M. L.; and Klunk, W. E. Synthesis and evaluation of
.sup.11C-labeled 6-substituted 2-arylbenzothiazoles as amyloid
imaging agents. J. Med. Chem. 2003, 46, 2740-2754.
[0140] 26. Klunk, W. E.; Wang, Y.; Huang, G. F.; Debnath, M. L.;
Holt, D. P.; Shao, L.; Hamilton, R. L.; Ikonomovic, M. D.; DeKosky,
S. T.; and Mathis, C. A. The binding of
2-(4'-methylaminophenyl)benzothiazole to postmortem brain
homogenates is dominated by the amyloid component. J. Neurosci.
2003, 23, 2086-2092.
[0141] 27. Klunk, W. E.; Wang, Y.; Huang, G. F.; Debnath, M. L.;
Holt, D. P.; and Mathis, C. A. Uncharged thioflavin-T derivatives
bind to amyloid-beta protein with high affinity and readily enter
the brain. Life Sci. 2001, 69, 1471-1484.
[0142] 28. Klunk, W. E.; Engler, H.; Nordberg, A.; Wang, Y.;
Blomqvist, G.; Holt, D. P.; Bergstrom, M.; Savitoheva, I.; Huang,
G. F.; Estrada, S.; Ausen, B.; Debnath, M. L.; Barletta, J.; Price,
J. C.; Sandell, J.; Lopresti, B. J.; Wall, A.; Koivisto, P.;
Antoni, G.; Mathis, C. A.; and Langstrom, B. Imaging brain amyloid
in Alzheimer's disease with Pittsburgh Compound-B. Ann. Neurol.
2004, 55, 306-319.
[0143] A further aspect of this invention is the novel synthesis of
aryl thioether derivatives of IMPY compounds by a rapid
palladium-catalyzed cross-coupling under microwave conditions. In
the following description, the cited references are identified
under Reference List No. 2.
[0144] Aromatic thioethers of IMPY compounds are valuable as
synthetic intermediates and as therapeutic drags..sup.1;2
Homogeneous catalysts based on either copper(I) or palladium have
been developed for their syntheses..sup.2;3 Mechanistic studies of
palladium-catalyzed aromatic substitution of halo or triflate
groups by thiolate show that both three and four coordinate
intermediates might be involved..sup.4-6 Indeed, both mono and
bidentate phosphine ligands have been developed for this type of
reaction..sup.7-12 Copper(I)-based batalysts for similar syntheses
are mechanistically much less clear..sup.13-17
[0145] In the synthesis of these compounds, introducing the
thioether group in the last step avoids manipulation and protection
of this sensitive group in the synthesis of the IMPY skeleton. The
aromatic halide substrates to be substituted are generally
accessible in multiple step syntheses..sup.18 Successful
substitution of aryl halides with thiolates requires the avoidance
of a competing reductive removal of the halogen substituent. The
objects of this aspect of the invention are to achieve (a) an
one-step introduction of a thioether group into a halogen position
in an aryl ring; (b) selective introduction of a thioether group at
an iodo position in the presence of other halogen substituents,
such as bromo; (c) tolerance of functional groups, especially amino
groups; and (d) fast microwave-assisted reaction conditions.
Disclosed herein is a general, efficient, and operationally simple
palladium-catalyzed aryl thioether synthesis that achieves these
objects.
[0146] Two reaction schemes are described below. The first is a
copper(I)-mediated coupling of aryl iodides with thiols (Scheme
B1), while the second is a palladium-catalyzed coupling of aryl
iodides with tin thiolates (Scheme B2).
##STR00016##
##STR00017##
[0147] The aryl halide 4-(6-iodoindolizin-2-yl)-N-methylbenzenamine
was used as the substrate, with either 2-mercaptoacetamide or
2-mercaptoethanol as the thiolating agent. In the copper-catalyzed
reaction (Scheme B1), the catalyst used was CuI. Two ligands,
ethylene glycol and ethylene diamine, were evaluated along with a
variety of bases and solvents (Table B1). Low conversion of the
iodo compound was observed with a catalytic amount of CuI. The
reaction went to completion only when a greater than stoichiometric
amount of CuI was used. For 2-mercaptoacetamide, the majority of
the reaction product arose from reductive removal of the iodo
group. For 2-mercaptoethanol, substitution of the iodo group was
nearly quantitative for one substrate when 5 equivalents of CuI
were used. However, no catalytic or general reaction was observed
and so no further effort was expended on this approach for other
substrates.
TABLE-US-00007 TABLE B1 The copper(I)-mediated coupling of aryl
iodides (1 or 2) with thiols: Percent yield of conversion followed
by ratio of substitution to reduction in parentheses. Method;
Substituents: (1) (2) (3) R.sup.2 = --CH.sub.2CONH.sub.2 100 81 R =
Me (1:4.0) (0:1) R.sup.2 = --CH.sub.2CONH.sub.2 81 100 100 R = H
(1:8.3) (1:4.1).sup.a,b (0:1) R.sup.2 = --CH.sub.2CH.sub.2OH 60 R =
Me (messy) R.sup.2 = --CH.sub.2CH.sub.2OH 96 R = H (1:0.038)
[0148] The general conditions were 130.degree. C., 10 min, 50 W,
300 p.s.i., substrate: thiol=1: 1.2-5. In Method (1), 1-5 equiv
CuI, 2 equiv ethylene diamine, 2 equiv t-BuOK or K.sub.2CO.sub.3 in
DMSO, and N-methylpyrrolidin-2-one (NMP) or dioxane were used. In
Method (2), 1-5 equiv CuI, 2 equiv ethylene glycol, and 2 equiv
t-BuOK in Py were used. In Method (3), 0.05 equiv CuI, and 2 equiv
Cs.sub.2CO.sub.3 in NMP were used. .sup.aWithout the thiol, the
reaction generated reduced and HOCH.sub.2CH.sub.2O-substituted
products. .sup.bNo reaction was observed when 1,2-dimethoxyethane
was used instead of ethylene glycol.
[0149] The palladium-catalyzed coupling of aryl iodides with thiols
(Scheme B2) was evaluated using the protocol originally established
by Buchwald et al,.sup.7 who used aryl bromides as substrates.
Variations here included the use of i) catalysts, such as
(DPPF)PdCl.sub.2; ii) catalyst precursors, such as
Pd.sub.2dba.sub.3 and Pd(OAc).sub.2; iii) ligands (L) as shown in
Scheme B2; iv) bases, such as NEt.sub.3, t-BuOK; and v) solvents,
such as NMP, dioxane, or toluene. Multiple products in similar
amounts were generated, including the desired substitution
Products. Given the success of a number of bulky monophosphines in
the catalysis of aryl C-N and C-O formation,.sup.6;19 a number of
bulky monophosphines were evaluated, as shown below:
##STR00018##
[0150] Under the general conditions used (150.degree. C., 10 min,
300 W, 300 p.s.i, with 0.05 equiv Pd.sub.2dba.sub.3, 0.1 equiv L,
2-4 equiv of NEt3 or t-BuOK, and ethanol or toluene as solvent),
the new ligands behave similarly to PPh.sub.3, giving less than 10%
total conversion of aryl iodide and ratios of substitution and
reduction ranging from 0.4 to 15. No further reaction progress was
observed on extended reaction time.
[0151] A number of thiolates have been used for the aromatic
substitution reaction..sup.1;20 When the tin thiolates were used in
Scheme B2, mainly substitution products accompanied by minor
reduction products were observed. Different kinds of bis-phosphine
ligands were evaluated, as shown below:
##STR00019##
[0152] The results are shown in Table B2.
TABLE-US-00008 TABLE B2 Scheme B2 with R.sup.1 = SnMe, R.sup.3 =
R.sup.4 = R.sup.5 = R.sup.6 = H, R.sup.7 = Me, X = I: The
palladium-catalyzed coupling of aryl iodides (1 or 2) with tin
thiolates. General conditions: 150.degree. C., 10 min, 300 W, 300
psi, substrate:thiolate = 1:1, in toluene. Results shown are
percent yield of conversion followed by ratio of substitution to
reduction in parentheses. R.sup.2; R.sup.6: CH.sub.2CONH.sub.2;
CH.sub.2CONH.sub.2; CH.sub.2CONH.sub.2; CH.sub.2CONH.sub.2; Ligand:
H Me H Me BINAP 17 (1:0.13).sup.a BINAP-Tol 81 (1:0.12).sup.b DPPF
100 (1:0).sup.c 56 (1:0.15).sup.d DiPPF 51 (1:0.019).sup.e 76
(1:0.14).sup.f 6 (1:0.083).sup.g .sup.a0.1 equiv
Pd.sub.2(dba).sub.3, 0.1 equiv L. .sup.b0.2 equiv
Pd.sub.2(dba).sub.3, 0.4 equiv L. .sup.c0.11 equiv
Pd.sub.2(dba).sub.3, 0.14 equiv L. .sup.d0.1 equiv
Pd.sub.2(dba).sub.3, 0.1 equiv L. .sup.e0.1 equiv
Pd.sub.2(dba).sub.3, 0.2 equiv L. .sup.f0.2 equiv
Pd.sub.2(dba).sub.3, 0.4 equiv L. .sup.g0.2 equiv
Pd.sub.2(dba).sub.3, 0.4 equiv L.
[0153] BINAP-Tol, DPPF, and DiPPF are efficient ligands for the
reaction. Since a number of chelating phosphines have been used in
enantioseleotive hydrogenation,.sup.21 they provide further
excellent candidates for evaluation in this reaction. The yields
and relative degrees of substitution and reduction for various
phosphines when used in Scheme B2 are shown below in Table B3. Once
again, ferrocene-based bis-phosphine,f-binaphane, provided the best
combination of reactivity and selectivity.
TABLE-US-00009 TABLE B3 Scheme B2 with R.sup.1 = SnMe, R.sup.2 =
CH.sub.2CONH.sub.2, R.sup.3 = R.sup.4 = R.sup.5 = R.sup.6 = H,
R.sup.7 = Me, X = I: The palladium-catalyzed coupling of aryl
iodide with tin thiolates. Ligands Products T-Phos 22 (1:0.91)
TangPhos 7 (1:0.71) Binapine 1 (1:0.63) f-Binaphane 11 (1:0.077)
C4TunePhos 8 (1:0.77) DuanPhos 0 Binaphane 0
General conditions; 150.degree. C., 10 min, 300W, 300 psi,
substrate:thiolate=1:1, 0.1 equiv Pd.sub.2(dba).sub.3 and 0.1
equiv. ligand in toluene. Results shown are percent yield of
conversion followed by ratio of substitution to reduction in
parentheses. The structures of the ligand are shown in Ref. 21.
[0154] Palladium catalysts based on phosphine oxide ligands are a
new type of efficient, versatile, air-stable and preformed
homogeneous catalysts for the C-S bond formation..sup.8-10 Ten of
the most common catalysts were evaluated in Scheme B2. The results
are shown in Table B4 which indicates that low reactivities and
selectivities were generally observed.
TABLE-US-00010 TABLE B4 The palladium-catalyzed coupling of aryl
iodide (2) with tin thiolates, using Scheme B2 with R.sup.1 = SnMe,
R.sup.2 = CH.sub.2CONH.sub.2, R.sup.3 = R.sup.4 = R.sup.5 = R.sup.6
= H, R.sup.7 = Me, X = I. General conditions: 150.degree. C., 10
min, 300 W, 300 psi, substrate:thiolate = 1:1.5, in toluene. In
Method (i), 2.5 equiv K.sub.2CO.sub.3 was used; in Method (ii), 2.5
equiv K.sub.2CO.sub.3 and 2.5 equiv TEA were used, and in Method
(iii), 2 equiv t-BuOK were used. The catalyst structures are shown
in Refs. 8-10. The results shown are percent yield of conversion
followed by ratio of substitution to reduction in parentheses.
Method: Catalyst: (i) (ii) (iii) POPd 0 0 PXPd 2 (1:0.63) PXPd7 7
(1:0.77) 1 (1:0.83) 3 (1:0.50) POPd2 0.sup.a,b 0 1 (1:0) PXPd2 2
(1:1.7) POPd6 0 PXPd6 2 (1:2.5) POPd1 0 0 0 POPd7 0 0 Ph1-Phoxide 3
(1:0.59) .sup.aWhen the thiol was used, no reaction was observed.
.sup.bOther thiols, such as HSCH.sub.2CH.sub.2OH, also did not
react.
[0155] When aryl bromides were used to compare the reactivity of
thiols vs. tin-thiolates, the thiols showed better reactivity but
less selectivity, as in reactions with the iodides. The results are
shown in Table B5.
TABLE-US-00011 TABLE B5 The palladium-catalyzed coupling of aryl
bromides with thiols or tin thiolates, using Scheme B2 with R.sup.3
= R.sup.4 = H, R.sup.5 = Me, R.sup.6 = H, R.sup.7 = Me, X = Br.
General conditions: 150.degree. C., 10 min, 300 W, 300 psi,
substrate:thiolate = 1:1.5, in toluene. The results shown are
percent yield of conversion, followed by ratio of substitution to
reduction in parentheses. Ligand: R.sup.1; R.sup.2: BINAP BINAP Tol
DPPF DiPPF R.sup.1 = SnMe.sub.3 6 (1:0).sup.a 16 (1:0).sup.b 0.4 14
(1:0).sup.d R.sup.2 = CH.sub.2CH.sub.2OH R.sup.1 = SnMe.sub.3 26
(1:0).sup.e R.sup.2 = CH.sub.2CONH.sub.2 R.sup.1 = H 100 (1:0.06)
R.sup.2 = CH.sub.2CH.sub.2OH R.sup.1 = H 26 (1:0.67).sup.h R.sup.2
= CH.sub.2CONH.sub.2 .sup.a0.1 equiv Pd.sub.2(dba).sub.3 and 0.1
equiv L. .sup.b0.2 equiv Pd.sub.2(dba).sub.3 and 0.2 equiv L.
.sup.c0.1 equiv Pd.sub.2(dba).sub.3 and 0.1 equiv L. .sup.d0.2
equiv Pd.sub.2(dba).sub.3 and 0.4 equiv L. .sup.e0.2 equiv
Pd.sub.2(dba).sub.3 and 0.4 equiv L. General conditions for thiol
reaction: 150.degree. C., 10 min, 300 W, 300 p.s.i, 11.2 equiv
t-BuOK; .sup.f0.2 equiv Pd(O.Ac).sub.2 and 0.23 equiv L in dioxane,
.sup.g0.2 equiv Pd.sub.2(dba).sub.3 and 0.23 equiv L in toluene,
.sup.h0.2 equiv Pd.sub.2(dba).sub.3 and 0.23 equiv L in
dioxane.
[0156] The combination of Pd.sub.2dba.sub.3 (dipalladium
dibenzylideneacetone) as catalyst, DiPPF as ligand and tin thiolate
as reagent was used to synthesize several aryl thiolates in
moderate to high yield. The results are shown in Table B6.
TABLE-US-00012 TABLE B6 The palladium-catalyzed synthesis of
thioethers with Pd.sub.2dba.sub.3 as catalyst, DiPPF as ligand, and
tin thiolate, using Scheme B2 with R.sup.1 = SnMe.sub.3, R.sup.3 =
R.sup.4 = H, R.sup.7 = Me, X, R.sup.2, R.sup.5, and R.sup.6 are as
shown in the table. General conditions: 150.degree. C., 10 min, 300
W, 300 psi, substrate:thiolate = 1:1-1.5, in toluene. In Method C1,
0.1 equiv Pd.sub.2(dba).sub.3 and 0.1 equiv DiPPF were used; in
Method C2, 0.2 equiv Pd.sub.2(dba).sub.3 and 0.4 equiv DiPPF were
used; in Method C3, 0.2 equiv Pd.sub.2(dba).sub.3 and 0.2 equiv
DiPPF were used; in Method C4, 0.2 equiv Pd.sub.2(dba).sub.3 and
0.8 equiv DiPPF were used. The results shown are the isolated yield
of the substitution product from reverse phase HPLC. X R.sup.5
R.sup.6 R.sup.2 Method Yield (%) I H H CH.sub.2CONH.sub.2 C1 78 I H
Me CH.sub.2CONH.sub.2 C2 85 I H H CH.sub.2CH.sub.2OH C1 91 I H Me
CH.sub.2CH.sub.2OH C2 89 I H Me CH.sub.2C.sub.6H.sub.4OCH.sub.3 C3
.sup. 85.sup.a Br Me H CH.sub.2CONH.sub.2 C4 69 Br Me H
CH.sub.2CH.sub.2OH C4 69 .sup.aWhen B.sub.2O.sub.3 was used with
the thiol instead of tin thiolate, no reaction was observed.
[0157] When bromo and iodo groups were both present in the same
compound, selective substitution of the iodo group was realized
using the same reaction conditions developed above, using
stoichiometric quantities of tin-thiolates. The results are shown
in Table B7.
TABLE-US-00013 TABLE B7 The palladium-catalyzed synthesis of
thioethers with Pd.sub.2(dba).sub.3 as catalyst, DiPPF as ligand,
and tin thiolate, using Scheme B2 with R.sup.1 = SnMe.sub.3,
R.sup.3R.sup.4 = CH.sub.2CH.sub.2, R.sup.5 = Br, R.sup.6 = Me, and
R.sup.7 = Ac, R.sup.2 is as shown in the table. General conditions:
150.degree. C., 10 min, 300 W, 300 psi, substrate:thiolate = 1:1,
0.1 equiv Pd.sub.2(dba).sub.3, 0.1 equiv DiPPF, in toluene. The
results shown are the isolated yield of the substitution product
from reverse phase HPLC. X R.sup.2 Yield (%) I CH.sub.2CONH.sub.2
60 I CH.sub.2CH.sub.2OH 94
[0158] The process method for the rapid, selective and efficient
substitution of bromo or iodo groups in aryl halides by
tin-thiolates with microwave heating, as disclosed herein, is
applicable to substrates with an easily reducible iodo group, in
either the presence or absence of a bromo group. Lower reactivity
and higher selectivity were observed as compared with those using
thiols as reagents. The corresponding reactions under conventional
heating did not generate any appreciable amount of products except
for those without any heteroatoms in the substrates (no data
shown).
Reference List No. 2
[0159] 1. Dickens, M. J.; Gilday, J. P.; Mowlem, T. J.; Widdowson,
D. A. Tetrahedron 1991, 47, 8621-8634.
[0160] 2. Kondo, T.; Mitsudo, T. Chem.Rev. 2000, 100,
3205-3220.
[0161] 3. Baranano, D.; Mann, G.; Hartwig, J. F. Curr. Org. Chem.
1997, 1, 187-305.
[0162] 4. Baranano, D.; Hartwig, J. F. J. Am. Chem. Soc. 1995, 117,
2937-2938.
[0163] 5. Louie, J.; Hartwig, J. F. J. Am. Chem. Soc. 1995, 117,
11598-11599.
[0164] 6. Hartwig, J. F. Acc. Chem. Res. 1998, 31, 852-860.
[0165] 7. Murata, M.; Buchwald, S. L. Tetrahedron 2004, 60,
7397-7403.
[0166] 8. Li, G. Y. J. Org. Chem. 2002, 67, 3643-3650.
[0167] 9. Li, G. Y.; Zheng, G.; Noonan, A. F. J. Org. Chem. 2001,
66, 8677-8681.
[0168] 10. Li, G. Y. Angew.Chem.Int.Ed. 2001, 40, 1513-1516.
[0169] 11. Schopfer, U.; Schlapbach, A. Tetrahedron 2001, 57,
3069-3073.
[0170] 12. Itoh, T.; Mase, T. Organic Letters 2004, 6,
4587-4590.
[0171] 13. Kwong, F. Y.; Buchwald, S. L. Organic Letters 2002, 4,
3517-3520.
[0172] 14. Bates, C. G.; Gujadhur, R. K.; Venkataraman, D. Organic
Letters 2002, 4, 2803-2806.
[0173] 15. Hickman, R. J. S.; Christie, B. J.; Guy, R. W.; White.
T. J. Austr. J. Chem. 1985, 38, 899-904.
[0174] 16. Palomo, C.; Oiarbide, M.; Lopez, R.; Gomez-Bengoa, E.
Tetrahedron Letters 2000, 41, 1283-1286.
[0175] 17. Wu, Y. J.; He, H. Synlett 2003, 1789-1790.
[0176] 18. Cai, L.; Chin, F. T.; Pike, V. W.; Toyama, H.; Liow, J.
S.; Zoghbi, S. S.; Modell, K.; Briard, E.; Shetty, H. U.; Sinclair,
K.; Donohue, S.; Tipre, D.; Kung, M. P.; Dagostin, C.; Widdowson,
D. A.; Green, M.; Gao, W.; Herman, M. M.; Ichise, M.; Innis, R. B.
J. Med Chem. 2004, 47, 2208-2218.
[0177] 19. Wolfe, J. P.; Wagaw, S.; Marcoux, J. -F.; Buchwald, S.
L. Acc. Chem. Res 1998, 31, 805-818.
[0178] 20. Ishiyama, T.; Mori, M.; Suzuki, A.; Miyaura, N. J.
Organomet. Chem. 1996, 525, 225-231.
[0179] 21. Tang, W.; Zhang; X. Chem. Rev. 2003, 103, 3029-3069.
* * * * *