U.S. patent application number 11/166358 was filed with the patent office on 2006-01-26 for gonadotropin-releasing hormone receptor antagonists and methods relating thereto.
This patent application is currently assigned to Neurocrine Biosciences, Inc.. Invention is credited to Chen Chen, Patrick J. Connors, Fabio Tucci, Yun-Fei Zhu.
Application Number | 20060019996 11/166358 |
Document ID | / |
Family ID | 23200303 |
Filed Date | 2006-01-26 |
United States Patent
Application |
20060019996 |
Kind Code |
A1 |
Tucci; Fabio ; et
al. |
January 26, 2006 |
Gonadotropin-releasing hormone receptor antagonists and methods
relating thereto
Abstract
GnPRH receptor antagonists are disclosed which have utility in
the treatment of a variety of sex-hormone related conditions in
both men and women. The compounds of this invention have the
structure: ##STR1## wherein A, R.sub.1, R.sub.2, R.sub.3a,
R.sub.3b, R.sub.4, R.sub.5, R.sub.6 R.sub.7 and n are as defined
herein, including stereoisomers, prodrugs and pharmaceutically
acceptable salts thereof. Also disclosed are compositions
containing a compound of this invention in combination with a
pharmaceutically acceptable carrier, as well as methods relating to
the use thereof for antagonizing gonadotropin-releasing hormone in
a subject in need thereof.
Inventors: |
Tucci; Fabio; (San Diego,
CA) ; Connors; Patrick J.; (Green Bay, WI) ;
Zhu; Yun-Fei; (San Diego, CA) ; Chen; Chen;
(San Diego, CA) |
Correspondence
Address: |
SEED INTELLECTUAL PROPERTY LAW GROUP PLLC
701 FIFTH AVE
SUITE 6300
SEATTLE
WA
98104-7092
US
|
Assignee: |
Neurocrine Biosciences,
Inc.
San Diego
CA
92130-1102
|
Family ID: |
23200303 |
Appl. No.: |
11/166358 |
Filed: |
June 24, 2005 |
Related U.S. Patent Documents
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Application
Number |
Filing Date |
Patent Number |
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10211978 |
Aug 2, 2002 |
6939883 |
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11166358 |
Jun 24, 2005 |
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60309937 |
Aug 2, 2001 |
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Current U.S.
Class: |
514/336 |
Current CPC
Class: |
A61P 37/02 20180101;
A61P 25/20 20180101; C07D 401/12 20130101; A61P 13/08 20180101;
C07D 473/24 20130101; C07D 409/14 20130101; A61P 15/00 20180101;
A61P 5/00 20180101; C07D 401/06 20130101; C07D 213/68 20130101;
C07D 405/12 20130101; A61P 1/00 20180101; A61P 43/00 20180101; C07D
453/02 20130101; C07D 409/04 20130101; C07D 211/86 20130101; A61P
17/00 20180101; A61P 5/04 20180101; A61P 35/00 20180101; C07D
453/06 20130101; C07D 213/69 20130101 |
Class at
Publication: |
514/336 |
International
Class: |
A61K 31/44 20060101
A61K031/44 |
Goverment Interests
STATEMENT OF GOVERNMENT INTEREST
[0002] Partial funding of the work described herein was provided by
the U.S. Government under Grant No. R43-HD38625 provided by the
National Institute of Health. The U.S. Government may have certain
rights in this invention.
Claims
1-5. (canceled)
6. A method for antagonizing gonadotropin-releasing hormone in a
subject in need thereof, comprising administering to the subject an
effective amount of a compound having the following structure:
##STR48## or a stereoisomer, prodrug or pharmaceutically acceptable
salt thereof, wherein: A is 0 or a bond; n is 1, 2, 3 or 4; R.sub.1
and R.sub.2 are the same or different and independently hydrogen
alky, substituted alkyl, aryl, substituted aryl, arylalkyl,
substituted arylalkyl, heterocycle, substituted heterocycle,
heterocyclealkyl, substituted heterocyclealkyl,
--C(R.sub.8)(.dbd.NR.sub.9) or
--C(NR.sub.10R.sub.11)(.dbd.NR.sub.9); or R.sub.1 and R.sub.2 taken
together with the nitrogen atom to which they are attached form a
heterocycle or a substituted heterocycle, R.sub.3a and R.sub.3b are
the same or different and, at each occurrence, independently
hydrogen, alkyl, substituted alkyl, alkoxy, alkylthio, alkylamino,
aryl, substituted aryl, arylalkyl, substituted arylalkyl,
heterocycle, substituted heterocycle, heterocyclealkyl, substituted
heterocyclealkyl, --COOR.sub.12 or --CONR.sub.10R.sub.11; or
R.sub.3a and R.sub.3b taken together with the carbon atom to which
they are attached form a homocycle, substituted homocycle,
heterocycle or substituted heterocycle, or R.sub.3a and the carbon
to which it is attached taken together with R.sub.1 and the
nitrogen to which it is attached form a heterocycle or substituted
heterocycle; R.sub.4 is hydrogen, alkyl or substituted alkyl;
R.sub.5 is arylalkyl, substituted arylalkyl, heteroarylalkyl or
substituted heteroarylalkyl; R.sub.6 is aryl, substituted aryl,
heteroaryl or substituted heteroaryl; R.sub.7 is hydrogen, alkyl or
substituted alkyl, R.sub.8, R.sub.9, R.sub.10 and R.sub.11 are the
same or different and, at each occurrence, independently hydrogen,
alkyl, substituted alkyl, aryl, substituted aryl, arylalkyl,
substituted arylalkyl heterocycle, substituted heterocycle,
heterocyclealkyl or substituted heterocyclealkyl; and R.sub.12 is
hydrogen, alkyl or substituted alkyl.
7. A method for treating a sex-hormone related condition of a
subject in need thereof, comprising administering to the subject an
effective amount of a compound having the following structure:
##STR49## or a stereoisomer, prodrug or pharmaceutically acceptable
salt thereof, wherein: A is 0 or a bond; n is 1, 2, 3 or 4; R.sub.1
and R.sub.2 are the same or different and independently hydrogen,
alkyl, substituted alkyl, aryl, substituted aryl, arylalkyl,
substituted arylalkyl, heterocycle, substituted heterocycle,
heterocyclealkyl, substituted heterocyclealkyl,
--C(R.sub.8)(.dbd.NR.sub.9) or
--C(NR.sub.10R.sub.11)(.dbd.NR.sub.9); or R.sub.1 and R.sub.2 taken
together with the nitrogen atom to which they are attached form a
heterocycle or a substituted heterocycle; R.sub.3a and R.sub.3b are
the same or different and, at each occurrence, independently
hydrogen, alkyl, substituted alkyl, alkoxy, alkylthio, alkylamino,
aryl, substituted aryl, arylalkyl, substituted arylalkyl,
heterocycle, substituted heterocycle, heterocyclealkyl, substituted
heterocyclealkyl, --COOR.sub.12 or --CONR.sub.10R.sub.11; or
R.sub.3a and R.sub.3b taken together with the carbon atom to which
they are attached form a homocycle, substituted homocycle,
heterocycle or substituted heterocycle, or R.sub.3a and the carbon
to which it is attached taken together with R.sub.1 and the
nitrogen to which it is attached form a heterocycle or substituted
heterocycle; R.sub.4 is hydrogen, alkyl or substituted alkyl;
R.sub.5 is arylalkyl, substituted arylalkyl, heteroarylalkyl or
substituted heteroarylalkyl; R.sub.6 is aryl, substituted aryl,
heteroaryl or substituted heteroaryl; R.sub.7 is hydrogen, alkyl or
substituted alkyl; R.sub.8, R.sub.9, R.sub.10 and R.sub.11 are the
same or different and, at each occurrence, independently hydrogen,
alkyl, substituted alkyl, aryl, substituted aryl, arylalkyl,
substituted arylalkyl, heterocycle, substituted heterocycle,
heterocyclealkyl or substituted heterocyclealkyl; and R.sub.12 is
hydrogen, alkyl or substituted alkyl.
8. The method of claim 7 wherein the sex-hormone related condition
is cancer, benign prostatic hypertrophy or myoma of the uterus.
9. The method of claim 8 wherein the cancer is prostatic cancer,
uterine cancer, breast cancer or pituitary gonadotroph
adenomas.
10. The method of claim 7 wherein the sex-hormone related condition
is endometriosis, polycystic ovarian disease, uterine fibroids or
precocious puberty.
11. A method for preventing pregnancy of a subject in need thereof,
comprising administering an effective amount of a compound having
the following structure: ##STR50## or a stereoisomer, prodrug or
pharmaceutically acceptable salt thereof, wherein: A is 0 or a
bond; n is 12, 3 or 4; R.sub.1 and R.sub.2 are the same or
different and independently hydrogen, alkyl substituted alkyl,
aryl, substituted aryl, arylalkyl, substituted arylalkyl,
heterocycle, substituted heterocycle, heterocyclealkyl, substituted
heterocyclealkyl, --C(R.sub.8)(.dbd.NR.sub.9) or
--C(NR.sub.10R.sub.11)(.dbd.NR.sub.9); or R.sub.1 and R.sub.2 taken
together with the nitrogen atom to which they are attached form a
heterocycle or a substituted heterocycle; R.sub.3a and R.sub.3b are
the same or different and, at each occurrence, independently
hydrogen, alkyl, substituted alkyl, alkoxy, alkylthio, alkylamino,
aryl, substituted aryl, arylalkyl, substituted arylalkyl,
heterocycle, substituted heterocycle, heterocyclealkyl, substituted
heterocyclealkyl, --COOR.sub.12 or --CONR.sub.10R.sub.11; or
R.sub.3a and R.sub.3b taken together with the carbon atom to which
they are attached form a homocycle, substituted homocycle,
heterocycle or substituted heterocycle; or R.sub.3a and the carbon
to which it is attached taken together with R.sub.1 and the
nitrogen to which it is attached form a heterocycle or substituted
heterocycle; R.sub.4 is hydrogen, alkyl or substituted alkyl;
R.sub.5 is arylalkyl, substituted arylalkyl, heteroarylalkyl or
substituted heteroarylalkyl; R.sub.6 is aryl, substituted aryl,
heteroaryl or substituted heteroaryl; R.sub.7 is hydrogen, alkyl or
substituted alkyl; R.sub.8, R.sub.9, R.sub.10 and R.sub.11 are the
same or different and, at each occurrence, independently hydrogen,
alkyl, substituted alkyl, aryl, substituted aryl, arylalkyl,
substituted arylalkyl, heterocycle, substituted heterocycle,
heterocyclealkyl or substituted heterocyclealkyl; and R.sub.12 is
hydrogen, alkyl or substituted alkyl.
12. A method for treating lupus erythematosis, irritable bowel
syndrome, premenstrual syndrome, hirsutism, short stature or sleep
disorders of a subject in need thereof, comprising administering to
the subject an effective amount of a compound having the following
structure: ##STR51## or a stereoisomer, prodrug or pharmaceutically
acceptable salt thereof. wherein: A is 0 or a bond; n is 1, 2, 3 or
4; R.sub.1 and R.sub.2 are the same or different and independently
hydrogen, alkyl, substituted alkyl, aryl, substituted aryl,
arylalkyl, substituted arylalkyl, heterocycle, substituted
heterocycle, heterocyclealkyl, substituted heterocyclealkyl,
--C(R.sub.8)(.dbd.NR.sub.9) or
--C(NR.sub.10R.sub.11)(.dbd.NR.sub.9); or R.sub.1 and R.sub.2 taken
together with the nitrogen atom to which they are attached form a
heterocycle or a substituted heterocycle; R.sub.3a and R.sub.3b are
the same or different and, at each occurrence, independently
hydrogen, alkyl, substituted alkyl, alkoxy, alkylthio, alkylamino,
aryl, substituted aryl, arylalkyl, substituted arylalkyl,
heterocycle, substituted heterocycle, heterocyclealkyl, substituted
heterocyclealkyl, --COOR.sub.12 or --CONR.sub.10R.sub.11; or
R.sub.3a and R.sub.3b taken together with the carbon atom to which
they are attached form a homocycle, substituted homocycle,
heterocycle or substituted heterocycle; or R.sub.3a and the carbon
to which it is attached taken together with R.sub.1 and the
nitrogen to which it is attached form a heterocycle or substituted
heterocycle; R.sub.4 is hydrogen, alkyl or substituted alkyl;
R.sub.5 is arylalkyl, substituted arylalkyl, heteroarylalkyl or
substituted heteroarylalkyl; R.sub.6 is aryl, substituted aryl,
heteroaryl or substituted heteroaryl; R.sub.7 is hydrogen, alkyl or
substituted alkyl; R.sub.8, R.sub.9, R.sub.10 and R.sub.11 are the
same or different and, at each occurrence, independently hydrogen,
alkyl, substituted alkyl aryl, substituted aryl, arylalkyl,
substituted arylalkyl, heterocycle, substituted heterocycle,
heterocyclealkyl or substituted heterocyclealkyl; and R.sub.12 is
hydrogen, alkyl or substituted alkyl.
Description
CROSS-REFERENCE TO RELATED APPLICATION
[0001] This application is a divisional of U.S. application Ser.
No. 10/211,978, filed Aug. 2, 2002, now allowed, which claims the
benefit of U.S. Provisional Patent Application No. 60/309,937,
filed Aug. 2, 2001, both of which applications are incorporated
herein by reference in their entireties.
BACKGROUND OF THE INVENTION
[0003] 1. Field of the Invention
[0004] This invention relates generally to gonadotropin-releasing
hormone (GnRH) receptor antagonists, and to methods of treating
disorders by administration of such antagonists to a warm-blooded
animal in need thereof.
[0005] 2. Description of the Related Art
[0006] Gonadotropin-releasing hormone (GnRH), also known as
luteinizing hormone-releasing hormone (LHRH), is a decapeptide
(pGlu-His-Trp-Ser-Tyr-Gly-Leu-Arg-Pro-Gly-NH.sub.2) that plays an
important role in human reproduction. GnRH is released from the
hypothalamus and acts on the pituitary gland to stimulate the
biosynthesis and release of luteinizing hormone (LH) and
follicle-stimulating hormone (FSH). LH released from the pituitary
gland is responsible for the regulation of gonadal steroid
production in both males and females, while FSH regulates
spermatogenesis in males and follicular development in females.
[0007] Due to its biological importance, synthetic antagonists and
agonists to GnRH have been the focus of considerable attention,
particularly in the context of prostate cancer, breast cancer,
endometriosis, uterine leiomyoma, and precocious puberty. For
example, peptidic GnRH agonists, such as leuprorelin
(pGlu-His-Trp-Ser-Tyr-D-Leu-Leu-Arg-Pro-NHEt), have been used to
treat such conditions. Such agonists appear to function by binding
to the GnRH receptor in the pituitary gonadotropins, thereby
inducing the synthesis and release of gonadotropins. Chronic
administration of GnRH agonists depletes gonadotropins and
subsequently down-regulates the receptor, resulting in suppression
of steroidal hormones after some period of time (e.g., on the order
of 2-3 weeks following initiation of chronic administration).
[0008] In contrast, GnRH antagonists are believed to suppress
gonadotropins from the onset, and thus have received the most
attention over the past two decades. To date, some of the primary
obstacles to the clinical use of such antagonists have been their
relatively low bioavailability and adverse side effects caused by
histamine release. However, several peptidic antagonists with low
histamine release properties have been reported, although they
still must be delivered via sustained delivery routes (such as
subcutaneous injection or intranasal spray) due to limited
bioavailability.
[0009] In view of the limitations associated with peptidic GnRH
antagonists, a number of nonpeptidic compounds have been proposed.
For example, Cho et al. (J. Med. Chem. 41:4190-4195, 1998)
discloses thieno[2,3-b]pyridin-4-ones for use as GnRH receptor
antagonists; U.S. Pat. Nos. 5,780,437 and 5,849,764 teach
substituted indoles as GnRH receptor antagonists (as do published
PCTs WO 97/21704, 98/55479, 98/55470, 98/55116, 98/55119, 97/21707,
97/21703 and 97/21435); published PCT WO 96/38438 discloses
tricyclic diazepines as GnRH receptor antagonists; published PCTs
WO97/14682, 97/14697 and 99/09033 disclose quinoline and
thienopyridine derivatives as GnRH antagonists; published PCTs WO
97/44037, 97/44041, 97/44321 and 97/44339 teach substituted
quinolin-2-ones as GnRH receptor antagonists; and published PCT WO
99/33831 discloses certain phenyl-substituted fused
nitrogen-containing bicyclic compounds as GnRH receptor
antagonists.
[0010] While significant strides have been made in this field,
there remains a need in the art for effective small molecule GnRH
receptor antagonists. There is also a need for pharmaceutical
compositions containing such GnRH receptor antagonists, as well as
methods relating to the use thereof to treat, for example,
sex-hormone related conditions. The present invention fulfills
these needs, and provides other related advantages.
BRIEF SUMMARY OF THE INVENTION
[0011] In brief, this invention is generally directed to
gonadotropin-releasing hormone (GnRH) receptor antagonists, as well
as to methods for their preparation and use, and to pharmaceutical
compositions containing the same. More specifically, the GnRH
receptor antagonists of this invention are compounds having the
following general structure (I): ##STR2## [0012] including
stereoisomers, prodrugs and pharmaceutically acceptable salts
thereof, wherein A, R.sub.1, R.sub.2, R.sub.3a, R.sub.3b, R.sub.4,
R.sub.5, R.sub.6, R.sub.7 and n are as defined below.
[0013] The GnRH receptor antagonists of this invention have utility
over a wide range of therapeutic applications, and may be used to
treat a variety of sex-hormone related conditions in both men and
women, as well as a mammal in general (also referred to herein as a
"subject"). For example, such conditions include endometriosis,
uterine fibroids, polycystic ovarian disease, hirsutism, precocious
puberty, gonadal steroid-dependent neoplasia such as cancers of the
prostate, breast and ovary, gonadotrophe pituitary adenomas, sleep
apnea, irritable bowel syndrome, premenstrual syndrome, benign
prostatic hypertrophy, contraception and infertility (e.g.,
assisted reproductive therapy such as in vitro fertilization). The
compounds of this invention are also useful as an adjunct to
treatment of growth hormone deficiency and short stature, and for
the treatment of systemic lupus erythematosis. The compounds are
also useful in combination with androgens, estrogens,
progesterones, and antiestrogens and antiprogestogens for the
treatment of endometriosis, fibroids, and in contraception, as well
as in combination with an angiotensin-converting enzyme inhibitor,
an angiotensin II-receptor antagonist, or a renin inhibitor for the
treatment of uterine fibroids. In addition, the compounds may be
used in combination with bisphosphonates and other agents for the
treatment and/or prevention of disturbances of calcium, phosphate
and bone metabolism, and in combination with estrogens,
progesterones and/or androgens for the prevention or treatment of
bone loss or hypogonadal symptoms such as hot flashes during
therapy with a GnRH antagonist.
[0014] The methods of this invention include administering an
effective amount of a GnRH receptor antagonist, preferably in the
form of a pharmaceutical composition, to a mammal in need thereof.
Thus, in still a further embodiment, pharmaceutical compositions
are disclosed containing one or more GnRH receptor antagonists of
this invention in combination with a pharmaceutically acceptable
carrier and/or diluent.
[0015] These and other aspects of the invention will be apparent
upon reference to the following detailed description. To this end,
various references are set forth herein which describe in more
detail certain background information, procedures, compounds and/or
compositions, and are each hereby incorporated by reference in
their entirety.
DETAILED DESCRIPTION OF THE INVENTION
[0016] As mentioned above, the present invention is directed
generally to compounds useful as gonadotropin-releasing hormone
(GnRH) receptor antagonists. The compounds of this invention have
the following structure (I): ##STR3## including stereoisomers,
prodrugs and pharmaceutically acceptable salts thereof, [0017]
wherein: [0018] A is O or a bond; [0019] n is 1, 2, 3 or 4; [0020]
R.sub.1 and R.sub.2 are the same or different and independently
hydrogen, alkyl, substituted alkyl, aryl, substituted aryl,
arylalkyl, substituted arylalkyl, heterocycle, substituted
heterocycle, heterocyclealkyl, substituted heterocyclealkyl,
--C(R.sub.8)(.dbd.NR.sub.9) or
--C(NR.sub.10R.sub.11)(.dbd.NR.sub.9); [0021] or R.sub.1 and
R.sub.2 taken together with the nitrogen atom to which they are
attached form a heterocycle or a substituted heterocycle; [0022]
R.sub.3a and R.sub.3b are the same or different and, at each
occurrence, independently hydrogen, alkyl, substituted alkyl,
alkoxy, alkylthio, alkylamino, aryl, substituted aryl, arylalkyl,
substituted arylalkyl, heterocycle, substituted heterocycle,
heterocyclealkyl, substituted heterocyclealkyl, --COOR.sub.12 or
--CONR.sub.10R.sub.11; [0023] or R.sub.3a and R.sub.3b taken
together with the carbon atom to which they are attached form a
homocycle, substituted homocycle, heterocycle or substituted
heterocycle; [0024] or R.sub.3a and the carbon to which it is
attached taken together with R.sub.1 and the nitrogen to which it
is attached form a heterocycle or substituted heterocycle; [0025]
R.sub.4 is hydrogen, alkyl or substituted alkyl; [0026] R.sub.5 is
arylalkyl, substituted arylalkyl, heteroarylalkyl or substituted
heteroarylalkyl; [0027] R.sub.6 is aryl, substituted aryl,
heteroaryl or substituted heteroaryl; [0028] R.sub.7 is hydrogen,
alkyl or substituted alkyl; [0029] R.sub.8, R.sub.9, R.sub.10 and
R.sub.11 are the same or different and, at each occurrence,
independently hydrogen, alkyl, substituted alkyl, aryl, substituted
aryl, arylalkyl, substituted arylalkyl, heterocycle, substituted
heterocycle, heterocyclealkyl or substituted heterocyclealkyl; and
[0030] R.sub.12 is hydrogen, alkyl or substituted alkyl.
[0031] As used herein, the above terms have the following
meaning:
[0032] "Alkyl" means a straight chain or branched, noncyclic or
cyclic, unsaturated or saturated aliphatic hydrocarbon containing
from 1 to 10 carbon atoms, while the term "lower alkyl" has the
same meaning as alkyl but contains from 1 to 6 carbon atoms. The
term "higher alkyl" has the same meaning as alkyl but contains from
2 to 10 carbon atoms. Representative saturated straight chain
alkyls include methyl, ethyl, n-propyl, n-butyl, n-pentyl, n-hexyl,
and the like; while saturated branched alkyls include isopropyl,
sec-butyl, isobutyl, tert-butyl, isopentyl, and the like.
Representative saturated cyclic alkyls include cyclopropyl,
cyclobutyl, cyclopentyl, cyclohexyl, and the like; while
unsaturated cyclic alkyls include cyclopentenyl and cyclohexenyl,
and the like. Cyclic alkyls are also referred to herein as a
"homocycles" or "homocyclic rings." Unsaturated alkyls contain at
least one double or triple bond between adjacent carbon atoms
(referred to as an "alkenyl" or "alkynyl," respectively).
Representative straight chain and branched alkenyls include
ethylenyl, propylenyl, 1-butenyl, 2-butenyl, isobutylenyl,
1-pentenyl, 2-pentenyl, 3-methyl-1-butenyl, 2-methyl-2-butenyl,
2,3-dimethyl-2-butenyl, and the like; while representative straight
chain and branched alkynyls include acetylenyl, propynyl,
1-butynyl, 2-butynyl, 1-pentynyl, 2-pentynyl, 3-methyl-1-butynyl,
and the like.
[0033] "Aryl" means an aromatic carbocyclic moiety such as phenyl
or naphthyl.
[0034] "Arylalkyl" means an alkyl having at least one alkyl
hydrogen atoms replaced with an aryl moiety, such as benzyl,
--(CH.sub.2).sub.2phenyl, --(CH.sub.2).sub.3phenyl,
--CH(phenyl).sub.2, and the like.
[0035] "Heteroaryl" means an aromatic heterocycle ring of 5- to 10
members and having at least one heteroatom selected from nitrogen,
oxygen and sulfur, and containing at least 1 carbon atom, including
both mono- and bicyclic ring systems. Representative heteroaryls
are furyl, benzofuranyl, thiophenyl, benzothiophenyl, pyrrolyl,
indolyl, isoindolyl, azaindolyl, pyridyl, quinolinyl,
isoquinolinyl, oxazolyl, isooxazolyl, benzoxazolyl, pyrazolyl,
imidazolyl, benzimidazolyl, thiazolyl, benzothiazolyl,
isothiazolyl, pyridazinyl, pyrimidinyl, pyrazinyl, triazinyl,
cinnolinyl, phthalazinyl, and quinazolinyl.
[0036] "Heteroarylalkyl" means an alkyl having at least one alkyl
hydrogen atom replaced with a heteroaryl moiety, such as
--CH.sub.2pyridinyl, --CH.sub.2pyrimidinyl, and the like.
[0037] "Heterocycle" (also referred to herein as a "heterocyclic
ring") means a 4- to 7-membered monocyclic, or 7- to 10-membered
bicyclic, heterocyclic ring which is either saturated, unsaturated,
or aromatic, and which contains from 1 to 4 heteroatoms
independently selected from nitrogen, oxygen and sulfur, and
wherein the nitrogen and sulfur heteroatoms may be optionally
oxidized, and the nitrogen heteroatom may be optionally
quaternized, including bicyclic rings in which any of the above
heterocycles are fused to a benzene ring. The heterocycle may be
attached via any heteroatom or carbon atom. Heterocycles include
heteroaryls as defined above. Thus, in addition to the heteroaryls
listed above, heterocycles also include morpholinyl,
pyrrolidinonyl, pyrrolidinyl, piperidinyl, hydantoinyl,
valerolactamyl, oxiranyl, oxetanyl, tetrahydrofuranyl,
tetrahydropyranyl, tetrahydropyridinyl, tetrahydroprimidinyl,
tetrahydrothiophenyl, tetrahydrothiopyranyl, tetrahydropyrimidinyl,
tetrahydrothiophenyl, tetrahydrothiopyranyl, and the like.
[0038] "Heterocyclealkyl" means an alkyl having at least one alkyl
hydrogen atom replaced with a heterocycle, such as
--CH.sub.2morpholinyl, and the like.
[0039] "Homocycle" (also referred to herein as "homocyclic ring")
means a saturated or unsaturated (but not aromatic) carbocyclic
ring containing from 3-7 carbon atoms, such as cyclopropane,
cyclobutane, cyclopentane, cyclohexane, cycloheptane, cyclohexene,
and the like.
[0040] The term "substituted" as used herein means any of the above
groups (i.e., alkyl, aryl, arylalkyl, heteroaryl, heteroarylalkyl,
homocycle, heterocycle and/or heterocyclealkyl) wherein at least
one hydrogen atom is replaced with a substituent. In the case of an
oxo substituent (".dbd.O") two hydrogen atoms are replaced. When
substituted one or more of the above groups are substituted,
"substituents" within the context of this invention include
halogen, hydroxy, oxo, cyano, nitro, amino, alkylamino,
dialkylamino, alkyl, alkoxy, alkylthio, haloalkyl, aryl, arylalkyl,
heteroaryl, heteroarylalkyl, heterocycle and heterocyclealkyl, as
well as --NR.sub.aR.sub.b, --NR.sub.aC(.dbd.O)R.sub.b,
--NR.sub.aC(.dbd.O)NR.sub.aNR.sub.b, --NR.sub.aC(.dbd.O)OR.sub.b
--NR.sub.aSO.sub.2R.sub.b, --C(.dbd.O)R.sub.a, --C(.dbd.O)OR.sub.a,
--C(.dbd.O)NR.sub.aR.sub.b, --OC(.dbd.O)NR.sub.aR.sub.b,
--OR.sub.a, --SR.sub.a, --SOR.sub.a, --S(.dbd.O).sub.2R.sub.a,
--OS(.dbd.O).sub.2R.sub.a and --S(.dbd.O).sub.2OR.sub.a. In
addition, the above substituents may be further substituted with
one or more of the above substituents, such that the substituent
substituted alky, substituted aryl, substituted arylalkyl,
substituted heterocycle or substituted heterocyclealkyl. R.sub.a
and R.sub.b in this context may be the same or different and
independently hydrogen, alkyl, haloalkyl, substituted alkyl, aryl,
substituted aryl, arylalkyl, substituted arylalkyl, heterocycle,
substituted heterocycle, heterocyclealkyl or substituted
heterocyclealkyl.
[0041] "Halogen" means fluoro, chloro, bromo and iodo.
[0042] "Haloalkyl" means an alkyl having at least one hydrogen atom
replaced with halogen, such as trifluoromethyl and the like.
[0043] "Alkoxy" means an alkyl moiety attached through an oxygen
bridge (i.e., --O-alkyl) such as methoxy, ethoxy, and the like.
[0044] "Alkylthio" means an alkyl moiety attached through a sulfur
bridge (i.e., --S-alkyl) such as methylthio, ethylthio, and the
like.
[0045] "Alkylsulfonyl" means an alkyl moiety attached through a
sulfonyl bridge (i.e., --SO.sub.2-alkyl) such as methylsulfonyl,
ethylsulfonyl, and the like.
[0046] "Alkylamino" and "dialkylamino" mean one or two alkyl moiety
attached through a nitrogen bridge (i.e., --N-alkyl) such as
methylamino, ethylamino, dimethylamino, diethylamino, and the
like.
[0047] With regard to the
"R.sub.1R.sub.2N(CR.sub.3aR.sub.3b).sub.n--" moiety of structure
(I), n may be 1, 2, 3 or 4. Accordingly, this moiety may be
represented by the following structure (i) when n is 1, (ii) when n
is 2, structure (iii) when n is 3, and structure (iv) when n is 4:
##STR4## wherein each occurrence of R.sub.3a and R.sub.3b above may
be the same or different, and are as defined above. For example,
when each occurrence of R.sub.3a and R.sub.3b in structures (i),
(ii) (iii) and (iv) is hydrogen, the
"R.sub.1R.sub.2N(CR.sub.3aR.sub.3b).sub.n--" moiety has the
structure R.sub.1R.sub.2N(CH.sub.2)--,
R.sub.1R.sub.2N(CH.sub.2).sub.2--,
R.sub.1R.sub.2N(CH.sub.2).sub.3-- and
R.sub.1R.sub.2N(CH.sub.2).sub.4--, respectively.
[0048] The compounds of the present invention may be prepared by
known organic synthesis techniques, including the methods described
in more detail in the Examples. However in general, the compounds
of structure (I) above may be made by the following Reaction
Schemes. All substituents in the following Reaction Schemes are as
defined above unless indicated otherwise. ##STR5##
[0049] Ketone or aldehyde 1 and an amine are dissolved neat or with
a cosolvent in an alkylorthoformate at a temperature of
0-50.degree. C. for 12 to 36 hours to give enamine 2. Compound 2 in
the presence of a substituted dioxinone in a protic solvent at
elevated or reflux temperature for 1-12 hours gives product 3.
Alternatively, compound 1 may be alkylated with a substituted acyl
halide in the presence of a base such as sodium hydride or
potassium t-butoxide in a solvent such as THF or dioxane for 1-24
hours at 0 to 50.degree. C. to give 4. 4 in the presence of a
substituted alkyl orthoester in a protic solvent at reflux for 1-24
hours gives 5. 5 and a primary amine, neat or in an inert solvent
such as DMF or DMSO, are heated at an elevated temperature or at
reflux temperature for 1-24 hours to give compound 3. (T. T.
Tidwell, J. Org. Chem., 1998, 63, 8636) ##STR6##
[0050] Compound 1 in the presence of a substituted aminocrotonate 6
is heated to 75-150.degree. C. in a solvent such as toluene to give
aminocrotonate 7 which cyclizes upon heating to give 8. (T.
Kametani, J. Heterocycl. Chem., 1977, 477) ##STR7##
[0051] Meldrum's acid 9 and a substituted acetyl halide in the
presence of a base such as TEA or pyridine in a solvent such as THF
or dichloromethane is stirred for 1-24 hours at a temperature of
20-75.degree. C. The reaction mix is evaporated and following a
quick workup, the residue is refluxed in an alcohol to give
.beta.-keto ester 10. Compound 10 in the presence of triazine or
DMFDMA followed by ammonium acetate gives compound 11. Use of a
primary amine in place of ammonium acetate gives 11 with a
substitution on the nitrogen. (M. Balogh, J. Heterocycl. Chem.,
1980, 359; R. Kiyama, Chem. Pharm. Bull., 1995, 43, 450)
##STR8##
[0052] Pyridone 12 is alkylated with an alkyl halide in the
presence of a base such as sodium hydride or potassium carbonate in
a solvent such as DMF or acetonitrile at 0 to 50.degree. C. for
1-24 hours to give alkylated compound 3. ##STR9##
[0053] Pyridone 13 may be halogenated using bromine/acetic acid,
ICI in chloroform, or NIS or NBS in a solvent such as chloroform at
0-50.degree. C. for 1-24 hours to 10 give 14. Compound 14 and an
appropriate boronic acid undergo a Suzuki coupling to give compound
3. ##STR10##
[0054] Pyridone ester 15 and a Grignard reagent at -78.degree. C.
in ether or THF for 1/4 to 4 hours, or using DIBAL-H at -78.degree.
C. for approximately 2 hours in THF or ether gives 16. Reductive
amination of 16 with a primary or secondary amine using a reagent
such as sodium cyanoborohydride or sodium triacetoxyborohydride at
0-50.degree. C. in an aprotic solvent such as methylene chloride
for 1-24 hours gives 17. Treatment of 16 with an appropriate
phosphonum salt and a base such as LDA or HMDS gives an enol that
is hydrolized by treatment with an aqueous acid such as HCl to give
18. Reductive amination with an appropriate amine gives compound
19.
[0055] Compounds of structure (I) may generally be referred to as
substituted 1H-pyridin-4-one compounds, representative compounds of
which include the following: [0056]
3-(2-Amino-2-phenyl-ethyl)-1-(2,6-difluoro-benzyl)-2,6-dimethyl-5-(2-fluo-
ro-3-methoxy-phenyl)-1H-pyridin-4-one; [0057]
3-(2-Amino-2-phenyl-ethyl)-1-(2,6-dichloro-benzyl)-2,6-dimethyl-5-(2-fluo-
ro-3-methoxy-phenyl)-1H-pyridin-4-one; [0058]
3-(2-Amino-2-phenyl-ethyl)-1-(2-fluoro-6-chloro-benzyl)-2,6-dimethyl-5-(2-
-fluoro-3-methoxy-phenyl)-1H-pyridin-4-one; [0059]
3-(2-Amino-2-phenyl-ethyl)-1-(2-fluoro-6-trifluoromethyl-benzyl)-2,6-dime-
thyl-5-(2-fluoro-3-methoxy-phenyl)-1H-pyridin-4-one; [0060]
3-(2-Amino-2-phenyl-ethyl)-1-(2-fluoro-6-methylsulfonyl-benzyl)-2,6-dimet-
hyl-5-(2-fluoro-3-methoxy-phenyl)-1H-pyridin-4-one; [0061]
3-(2-Amino-2-phenyl-ethyl)-1-(2-fluoro-benzyl)-2,6-dimethyl-5-(2-fluoro-3-
-methoxy-phenyl)-1H-pyridin-4-one; [0062]
3-(2-Amino-2-phenyl-ethyl)-1-(2-chloro-benzyl)-2,6-dimethyl-5-(2-fluoro-3-
-methoxy-phenyl)-1H-pyridin-4-one; [0063]
3-(2-Amino-2-phenyl-ethyl)-1-(2-trifluoromethyl-benzyl)-2,6-dimethyl-5-(2-
-fluoro-3-methoxy-phenyl)-1H-pyridin-4-one; [0064]
3-(2-Amino-2-phenyl-ethyl)-1-(2-methylsulfonyl-benzyl)-2,6-dimethyl-5-(2--
fluoro-3-methoxy-phenyl)-1H-pyridin-4-one; [0065]
3-(2-Amino-2-phenyl-ethyl)-1-(2,6-difluoro-benzyl)-2,6-dimethyl-5-(2-fluo-
ro-phenyl)-1H-pyridin-4-one; [0066]
3-(2-Amino-2-phenyl-ethyl)-1-(2,6-dichloro-benzyl)-2,6-dimethyl-5-(2-fluo-
ro-phenyl)-1H-pyridin-4-one; [0067]
3-(2-Amino-2-phenyl-ethyl)-1-(2-fluoro-6-chloro-benzyl)-2,6-dimethyl-5-(2-
-fluoro-phenyl)-1H-pyridin-4-one; [0068]
3-(2-Amino-2-phenyl-ethyl)-1-(2-fluoro-6-trifluoromethyl-benzyl)-2,6-dime-
thyl-5-(2-fluoro-phenyl)-1H-pyridin-4-one; [0069]
3-(2-Amino-2-phenyl-ethyl)-1-(2-fluoro-6-methylsulfonyl-benzyl)-2,6-dimet-
hyl-5-(2-fluoro-phenyl)-1H-pyridin-4-one; [0070]
3-(2-Amino-2-phenyl-ethyl)-1-(2-fluoro-benzyl)-2,6-dimethyl-5-(2-fluoro-p-
henyl)-1H-pyridin-4-one; [0071]
3-(2-Amino-2-phenyl-ethyl)-1-(2-chloro-benzyl)-2,6-dimethyl-5-(2-fluoro-p-
henyl)-1H-pyridin-4-one; [0072]
3-(2-Amino-2-phenyl-ethyl)-1-(2-trifluoromethyl-benzyl)-2,6-dimethyl-5-(2-
-fluoro-phenyl)-1H-pyridin-4-one; [0073]
3-(2-Amino-2-phenyl-ethyl)-1-(2-methylsulfonyl-benzyl)-2,6-dimethyl-5-(2--
fluoro-phenyl)-1H-pyridin-4-one; [0074]
3-(2-Amino-2-phenyl-ethyl)-1-(2,6-difluoro-benzyl)-2,6-dimethyl-5-(2-chlo-
ro-phenyl)-1H-pyridin-4-one; [0075]
3-(2-Amino-2-phenyl-ethyl)-1-(2,6-dichloro-benzyl)-2,6-dimethyl-5-(2-chlo-
ro-phenyl)-1H-pyridin-4-one; [0076]
3-(2-Amino-2-phenyl-ethyl)-1-(2-fluoro-6-chloro-benzyl)-2,6-dimethyl-5-(2-
-chloro-phenyl)-1H-pyridin-4-one; [0077]
3-(2-Amino-2-phenyl-ethyl)-1-(2-fluoro-6-trifluoromethyl-benzyl)-2,6-dime-
thyl-5-(2-chloro-phenyl)-1H-pyridin-4-one; [0078]
3-(2-Amino-2-phenyl-ethyl)-1-(2-fluoro-6-methylsulfonyl-benzyl)-2,6-dimet-
hyl-5-(2-chloro-phenyl)-1H-pyridin-4-one; [0079]
3-(2-Amino-2-phenyl-ethyl)-1-(2-fluoro-benzyl)-2,6-dimethyl-5-(2-chloro-p-
henyl)-1H-pyridin-4-one; [0080]
3-(2-Amino-2-phenyl-ethyl)-1-(2-chloro-benzyl)-2,6-dimethyl-5-(2-chloro-p-
henyl)-1H-pyridin-4-one; [0081]
3-(2-Amino-2-phenyl-ethyl)-1-(2-trifluoromethyl-benzyl)-2,6-dimethyl-5-(2-
-chloro-phenyl)-1H-pyridin-4-one; [0082]
3-(2-Amino-2-phenyl-ethyl)-1-(2-methylsulfonyl-benzyl)-2,6-dimethyl-5-(2--
chloro-phenyl)-1H-pyridin-4-one; [0083]
3-(2-Amino-2-phenyl-ethyl)-1-(2,6-difluoro-benzyl)-2,6-dimethyl-5-(3-meth-
oxy-phenyl)-1H-pyridin-4-one; [0084]
3-(2-Amino-2-phenyl-ethyl)-1-(2,6-dichloro-benzyl)-2,6-dimethyl-5-(3-meth-
oxy-phenyl)-1H-pyridin-4-one; [0085]
3-(2-Amino-2-phenyl-ethyl)-1-(2-fluoro-6-chloro-benzyl)-2,6-dimethyl-5-(3-
-methoxy-phenyl)-1H-pyridin-4-one; [0086]
3-(2-Amino-2-phenyl-ethyl)-1-(2-fluoro-6-trifluoromethyl-benzyl)-2,6-dime-
thyl-5-(3-methoxy-phenyl)-1H-pyridin-4-one; [0087]
3-(2-Amino-2-phenyl-ethyl)-1-(2-fluoro-6-methylsulfonyl-benzyl)-2,6-dimet-
hyl-5-(3-methoxy-phenyl)-1H-pyridin-4-one; [0088]
3-(2-Amino-2-phenyl-ethyl)-1-(2-fluoro-benzyl)-2,6-dimethyl-5-(3-methoxy--
phenyl)-1H-pyridin-4-one; [0089]
3-(2-Amino-2-phenyl-ethyl)-1-(2-chloro-benzyl)-2,6-dimethyl-5-(3-methoxy--
phenyl)-1H-pyridin-4-one; [0090]
3-(2-Amino-2-phenyl-ethyl)-1-(2-trifluoromethyl-benzyl)-2,6-dimethyl-5-(3-
-methoxy-phenyl)-1H-pyridin-4-one; [0091]
3-(2-Amino-2-phenyl-ethyl)-1-(2-methylsulfonyl-benzyl)-2,6-dimethyl-5-(3--
methoxy-phenyl)-1H-pyridin-4-one; [0092]
3-(2-Amino-2-phenyl-ethyl)-1-(2,6-difluoro-benzyl)-2,6-dimethyl-5-(2-chlo-
ro-3-methoxy-phenyl)-1H-pyridin-4-one; [0093]
3-(2-Amino-2-phenyl-ethyl)-1-(2,6-dichloro-benzyl)-2,6-dimethyl-5-(2-chlo-
ro-3-methoxy-phenyl)-1H-pyridin-4-one; [0094]
3-(2-Amino-2-phenyl-ethyl)-1-(2-fluoro-6-chloro-benzyl)-2,6-dimethyl-5-(2-
-chloro-3-methoxy-phenyl)-1H-pyridin-4-one; [0095]
3-(2-Amino-2-phenyl-ethyl)-1-(2-fluoro-6-trifluoromethyl-benzyl)-2,6-dime-
thyl-5-(2-chloro-3-methoxy-phenyl)-1H-pyridin-4-one; [0096]
3-(2-Amino-2-phenyl-ethyl)-1-(2-fluoro-6-methylsulfonyl-benzyl)-2,6-dimet-
hyl-5-(2-chloro-3-methoxy-phenyl)-1H-pyridin-4-one; [0097]
3-(2-Amino-2-phenyl-ethyl)-1-(2-fluoro-benzyl)-2,6-dimethyl-5-(2-chloro-3-
-methoxy-phenyl)-1H-pyridin-4-one; [0098]
3-(2-Amino-2-phenyl-ethyl)-1-(2-chloro-benzyl)-2,6-dimethyl-5-(2-chloro-3-
-methoxy-phenyl)-1H-pyridin-4-one; [0099]
3-(2-Amino-2-phenyl-ethyl)-1-(2-trifluoromethyl-benzyl)-2,6-dimethyl-5-(2-
-chloro-3-methoxy-phenyl)-1H-pyridin-4-one; and
[0100]
3-(2-Amino-2-phenyl-ethyl)-1-(2-methylsulfonyl-benzyl)-2,6-dimethy-
l-5-(2-chloro-3-methoxy-phenyl)-1H-pyridin-4-one.
[0101] In addition, representative compounds of the present
invention also include those compounds where the primary amine of
the above named compounds is substituted with a substituted alkyl
group or a cycloalkyl group. As described in the examples, one
method of alkylating amines and amides is by reductive alkylation.
There are many alternative methods well known in the chemical arts
for accomplishing the reductive alkylation procedure, and there are
many alternative alkylation methods. When an aldehyde, ketone,
carboxylic acid, or acid chloride is treated with a primary or
secondary amine in the presence of a reducing agent reductive
alkylation may take place. Suitable reducing agents include (but
are not limited to) sodium borohydride, sodium
triacetoxyborohydride, sodium cyanoborohydride, hydrogen gas and a
hydrogenation catalyst, zinc and hydrochloric acid, iron
pentacarbonyl and alcoholic potassium hydroxide, formic acid,
pyridine borohydride. Amines and amides may also be alkylated by
the reaction of formaldehyde and a Mannich base or by the
nucleophilic displacement of an alkyl halide or other leaving
groups. As an example, the Mitsunobu reaction allows the alkylation
of amines with primary or secondary alcohols and carboxylic acids
by activation of the hydroxyl group with triphenylphosphine to form
the leaving group triphenylphoshine oxide. Other commonly used
alkylation methods are described in March, Advanced Organic
Chemistry, 4th Ed., pp 1276-1277 (1992).
[0102] The compounds of the present invention may generally be
utilized as the free acid or free base. Alternatively, the
compounds of this invention may be used in the form of acid or base
addition salts. Acid addition salts of the free amino compounds of
the present invention may be prepared by methods well known in the
art, and may be formed from organic and inorganic acids. Suitable
organic acids include maleic, fumaric, benzoic, ascorbic, succinic,
methanesulfonic, acetic, trifluoroacetic, oxalic, propionic,
tartaric, salicylic, citric, gluconic, lactic, mandelic, cinnamic,
aspartic, stearic, palmitic, glycolic, glutamic, and
benzenesulfonic acids. Suitable inorganic acids include
hydrochloric, hydrobromic, sulfuric, phosphoric, and nitric acids.
Base addition salts included those salts that form with the
carboxylate anion and include salts formed with organic and
inorganic cations such as those chosen from the alkali and alkaline
earth metals (for example, lithium, sodium, potassium, magnesium,
barium and calcium), as well as the ammonium ion and substituted
derivatives thereof (for example, dibenzylammonium, benzylammonium,
2-hydroxyethylammonium, and the like). Thus, the term
"pharmaceutically acceptable salt" of structure (I) is intended to
encompass any and all acceptable salt forms.
[0103] In addition, prodrugs are also included within the context
of this invention. Prodrugs are any covalently bonded carriers that
release a compound of structure (I) in vivo when such prodrug is
administered to a patient. Prodrugs are generally prepared by
modifying functional groups in a way such that the modification is
cleaved, either by routine manipulation or in vivo, yielding the
parent compound. Prodrugs include, for example, compounds of this
invention wherein hydroxy, amine or sulfhydryl groups are bonded to
any group that, when administered to a patient, cleaves to form the
hydroxy, amine or sulfhydryl groups. Thus, representative examples
of prodrugs include (but are not limited to) acetate, formate and
benzoate derivatives of alcohol and amine functional groups of the
compounds of structure (I). Further, in the case of a carboxylic
acid (--COOH), esters may be employed, such as methyl esters, ethyl
esters, and the like.
[0104] With regard to stereoisomers, the compounds of structure (I)
may have chiral centers and may occur as racemates, racemic
mixtures and as individual enantiomers or diastereomers. Compounds
of structure (I) may also possess axial chirality, which may result
in atropisomers. All such isomeric forms are included within the
present invention, including mixtures thereof. Furthermore, some of
the crystalline forms of the compounds of structure (I) may exist
as polymorphs, which are included in the present invention. In
addition, some of the compounds of structure (I) may also form
solvates with water or other organic solvents. Such solvates are
similarly included within the scope of this invention.
[0105] The effectiveness of a compound as a GnRH receptor
antagonist may be determined by various assay methods. Suitable
GnRH antagonists of this invention are capable of inhibiting the
specific binding of GnRH to its receptor and antagonizing
activities associated with GnRH. For example, inhibition of GnRH
stimulated LH release in immature rats may be measured according to
the method of Vilchez-Martinez (Endocrinology 96:1130-1134, 1975).
Briefly, twenty-five day old male Spraque-Dawley rats are
administered an GnRH antagonist in saline or other suitable
formulation by oral gavage, sub-cutaneous injection, or intravenous
injection. This is followed by subcutaneous injection of 200 ng
GnRH in 0.2 ml saline. Thirty minutes after the last injection, the
animals are decapitated and trunk blood collected. After
centrifugation, the separated plasma is stored at -20.degree. C.
until determination of the LH and FSH by radioimmunoassay. Other
techniques for determining the activity of GnRH receptor
antagonists are well known in the field, such as the use of
cultured pituitary cells for measuring GnRH activity (Vale et al.,
Endocrinology 91:562-572, 1972), and a technique for measuring
radioligand binding to rat pituitary membranes (Perrin et al., Mol.
Pharmacol 23:44-51, 1983).
[0106] For example, effectiveness of a compound as a GnRH receptor
antagonist may be determined by one or more of the following
assays.
Rat Anterior Pituitary Cell Culture Assay of GnRH Antagonists
[0107] Anterior pituitary glands are collected from 7-week-old
female Sprague-Dawley rats and the harvested glands digested with
collagenase in a dispersion flask for 1.5 hr at 37.degree. C. After
collagenase digestion, the glands are further digested with
neuraminidase for 9 min at 37.degree. C. The digested tissue is
then washed with 0.1% BSA/McCoy's 5A medium, and the washed cells
suspended in 3% FBS/0.1 BSA/McCoy's 5A medium and plated into
96-well tissue culture plates at a cell density of 40,000 cells per
well in 200 .mu.l medium. The cells are then incubated at
37.degree. C. for 3 days. One pituitary gland normally yields one
96-well plate of cells, which can be used for assaying three
compounds. For assay of an GnRH antagonist, the incubated cells are
first washed with 0.1% BSA/McCoy's 5A medium once, followed by
addition of the test sample plus 1 nM GnRH in 200 .mu.l 0.1%
BSA/McCoy's 5A medium in triplicate wells. Each sample is assayed
at 5-dose levels to generate a dose-response curve for
determination of its potency on the inhibition of GnRH stimulated
LH and/or FSH release. After 4-hr incubation at 37.degree. C., the
medium is harvested and the level of LH and/or FSH secreted into
the medium determined by RIA.
RIA of LH and FSH
[0108] For determination of the LH levels, each sample medium is
assayed in duplicates and all dilutions are done with RIA buffer
(0.01M sodium phosphate buffer/0.15M NaCl/1% BSA/0.01% NaN3, pH
7.5) and the assay kit is obtained from the Nation Hormone and
Pituitary Program supported by NIDDK. To a 12.times.75 mm
polyethylene test tube is added 100 .mu.l of sample medium diluted
1:5 or rLH standard in RIA buffer and 100 .mu.l of [125I]-labeled
rLH (-30,000 cpm) plus 100 .mu.l of rabbit anti-rLH antibody
diluted 1:187,500 and 100 .mu.l RIA buffer. The mixture is
incubated at room temperature over-night. In the next day, 100
.mu.l of goat anti-rabbit IgG diluted 1:20 and 100 .mu.l of normal
rabbit serum diluted 1:1000 are added and the mixture incubated for
another 3 hr at room temperature. The incubated tubes are then
centrifuged at 3,000 rpm for 30 min and the supernatant removed by
suction. The remaining pellet in the tubes is counted in a
gamma-counter. RIA of FSH is done in a similar fashion as the assay
for LH with substitution of the LH antibody by the FSH antibody
diluted 1:30,000 and the labeled rLH by the labeled rFSH.
Radio-Iodination of GnRH Peptide
[0109] The GnRH analog is labeled by the chloramine-T method. To 10
.mu.g of peptide in 20 til of 0.5M sodium phosphate buffer, pH 7.6,
is added 1 mCi of Na125I, followed by 22.5 .mu.g chloramine-T and
the mixture vortexed for 20 sec. The reaction is stopped by the
addition of 60 .mu.g sodium metabisulfite and the free iodine is
removed by passing the iodinated mixture through a C-8 Sep-Pak
cartridge (Millipore Corp., Milford, MA). The peptide is eluted
with a small volume of 80% acetonitrile/water. The recovered
labeled peptide is further purified by reverse phase HPLC on a
Vydac C-18 analytical column (The Separations Group, Hesperia, CA)
on a Beckman 334 gradient HPLC system using a gradient of
acetonitrile in 0.1% TFA. The purified radioactive peptide is
stored in 0.1% BSA/20% acetonitrile/0.1% TFA at -80.degree. C. and
can be used for up to 4 weeks.
GnRH Receptor Membrane Binding Assay
[0110] Cells stably, or transiently, transfected with GnRH receptor
expression vectors are harvested, resuspended in 5% sucrose and
homogenized using a polytron homogenizer (2.times.15 sec). Nucleii
are removed by centrifugation (3000.times.g for 5 min.), and the
supernatant centrifuged (20,000.times.g for 30 min, 4.degree. C.)
to collect the membrane fraction. The final membrane preparation is
resuspended in binding buffer (10 mM Hepes (pH 7.5), 150 mM NaCl,
and 0.1% BSA) and stored at -70.degree. C. Binding reactions are
performed in a Millipore MultiScreen 96-well filtration plate
assembly with polyethylenimine coated GF/C membranes. The reaction
is initiated by adding membranes (40 ug protein in 130 ul binding
buffer) to 50 ul of [.sup.125I]-labeled GnRH peptide (100,000 cpm),
and 20 ul of competitor at varying concentrations. The reaction is
terminated after 90 minutes by application of vacuum and washing
(2.times.) with phosphate buffered saline. Bound radioactivity is
measured using 96-well scintillation counting (Packard Topcount) or
by removing the filters from the plate and direct gamma counting.
K.sub.i values are calculated from competition binding data using
non-linear least squares regression using the Prism software
package (GraphPad Software).
[0111] Activity of GnRH receptor antagonists are typically
calculated from the IC.sub.50 as the concentration of a compound
necessary to displace 50% of the radiolabeled ligand from the GnRH
receptor, and is reported as a "K.sub.i" value calculated by the
following equation: K i = IC 50 1 + L / K D ##EQU1## where
L=radioligand and K.sub.D=affinity of radioligand for receptor
(Cheng and Prusoff, Biochem. Pharmacol. 22:3099, 1973). GnRH
receptor antagonists of this invention have a K.sub.i of 100 .mu.M
or less. In a preferred embodiment of this invention, the GnRH
receptor antagonists have a K.sub.i of less than 10 .mu.M, and more
preferably less than 1 .mu.M, and even more preferably less than
0.1 .mu.M (i.e., 100 nM).
[0112] As mentioned above, the GnRH receptor antagonists of this
invention have utility over a wide range of therapeutic
applications, and may be used to treat a variety of sex-hormone
related conditions in both men and women, as well as mammals in
general. For example, such conditions include endometriosis,
uterine fibroids, polycystic ovarian disease, hirsutism, precocious
puberty, gonadal steroid-dependent neoplasia such as cancers of the
prostate, breast and ovary, gonadotrophe pituitary adenomas, sleep
apnea, irritable bowel syndrome, premenstrual syndrome, benign
prostatic hypertrophy, contraception and infertility (e.g.,
assisted reproductive therapy such as in vitro fertilization).
[0113] The compounds of this invention are also useful as an
adjunct to treatment of growth hormone deficiency and short
stature, and for the treatment of systemic lupus erythematosis.
[0114] In addition, the compounds are useful in combination with
androgens, estrogens, progesterones, and antiestrogens and
antiprogestogens for the treatment of endometriosis, fibroids, and
in contraception, as well as in combination with an
angiotensin-converting enzyme inhibitor, an angiotensin II-receptor
antagonist, or a renin inhibitor for the treatment of uterine
fibroids. The compounds may also be used in combination with
bisphosphonates and other agents for the treatment and/or
prevention of disturbances of calcium, phosphate and bone
metabolism, and in combination with estrogens, progesterones and/or
androgens for the prevention or treatment of bone loss or
hypogonadal symptoms such as hot flashes during therapy with a GnRH
antagonist.
[0115] In another embodiment of the invention, pharmaceutical
compositions containing one or more GnRH receptor antagonists are
disclosed. For the purposes of administration, the compounds of the
present invention may be formulated as pharmaceutical compositions.
Pharmaceutical compositions of the present invention comprise a
GnRH receptor antagonist of the present invention and a
pharmaceutically acceptable carrier and/or diluent. The GnRH
receptor antagonist is present in the composition in an amount
which is effective to treat a particular disorder--that is, in an
amount sufficient to achieve GnRH receptor antagonist activity, and
preferably with acceptable toxicity to the patient. Typically, the
pharmaceutical compositions of the present invention may include a
GnRH receptor antagonist in an amount from 0.1 mg to 250 mg per
dosage depending upon the route of administration, and more
typically from 1 mg to 60 mg. Appropriate concentrations and
dosages can be readily determined by one skilled in the art.
[0116] Pharmaceutically acceptable carrier and/or diluents are
familiar to those skilled in the art. For compositions formulated
as liquid solutions, acceptable carriers and/or diluents include
saline and sterile water, and may optionally include antioxidants,
buffers, bacteriostats and other common additives. The compositions
can also be formulated as pills, capsules, granules, or tablets
which contain, in addition to a GnRH receptor antagonist, diluents,
dispersing and surface active agents, binders, and lubricants. One
skilled in this art may further formulate the GnRH receptor
antagonist in an appropriate manner, and in accordance with
accepted practices, such as those disclosed in Remington's
Pharmaceutical Sciences, Gennaro, Ed., Mack Publishing Co., Easton,
Pa. 1990.
[0117] In another embodiment, the present invention provides a
method for treating sex-hormone related conditions as discussed
above. Such methods include administering of a compound of the
present invention to a warm-blooded animal in an amount sufficient
to treat the condition. In this context, "treat" includes
prophylactic administration. Such methods include systemic
administration of a GnRH receptor antagonist of this invention,
preferably in the form of a pharmaceutical composition as discussed
above. As used herein, systemic administration includes oral and
parenteral methods of administration. For oral administration,
suitable pharmaceutical compositions of GnRH receptor antagonists
include powders, granules, pills, tablets, and capsules as well as
liquids, syrups, suspensions, and emulsions. These compositions may
also include flavorants, preservatives, suspending, thickening and
emulsifying agents, and other pharmaceutically acceptable
additives. For parental administration, the compounds of the
present invention can be prepared in aqueous injection solutions
which may contain, in addition to the GnRH receptor antagonist,
buffers, antioxidants, bacteriostats, and other additives commonly
employed in such solutions.
[0118] The following example is provided for purposes of
illustration, not limitation. In summary, the GnRH receptor
antagonists of this invention may be assayed by the general methods
disclosed above, while the following Examples disclose the
synthesis of representative compounds of this invention.
EXAMPLE 1
SYNTHESIS OF
1-(2,6-DIFLUOROBENZYL)-2,6-DIMETHYL-3-BROMO-5-ETHOXYCARBONYL-4-PYRIDONE
[0119] ##STR11##
Step 1A Ethyl-3-(2.6-difluorobenzylamino)crotonate:
[0120] 2,6-Difluorobenzylamine (5.00 g, 35.0 mmol) was added to
trimethyl orthoformate (88.0 mL) under N.sub.2. The resulting
solution was stirred at room temperature for 20 minutes. Ethyl
acetoacetate (4.5 mL, 35.0 mmol) was then added dropwise and the
resulting mixture stirred for 24 hours (reaction progress monitored
by LC/MS). The volatiles were removed under vacuum and the product
solidified upon standing. Yield 7.88 g (30.9 mmol, 88%). .sup.1H
NMR (CDCl.sub.3, 300 MHz): 8.86 (br, 1H), 7.30-7.21 (m, 1H),
6.94-6.81 (m, 2H), 4.48 (s, 2H), 4.07 (q, J=6.9 Hz, 2H), 2.03 (s,
3H), 1.23 (t, J=6.9 Hz, 3H); LRMS m/z 256.1 (M.sup.++1).
Step 1B
1-(2.6-Difluorobenzyl)-2,6-dimethyl-3-ethoxycarbonyl-4-pyridone:
[0121] A mixture of ethyl-3-(2,6-difluorobenzylamino)crotonate
(2.55 g, 10.0 mmol) and 2,2,6-trimethyl-1,3-dioxin-4-one (1.85 g,
13.0 mmol) in toluene (100 mL) was heated at reflux for 3 hours.
The reaction progress was monitored by LC/MS. An additional aliquot
of 2,2,6-trimethyl-1,3-dioxin-4-one (2.0 mL) was added and the
reaction mixture was refluxed for another 2 hours. LC/MS indicated
complete conversion to product. The mixture was cooled down and the
solvent was removed in vacuo. The solid residue was triturated with
diethyl ether to yield the product as a tan solid (1.37 g, 4.3
mmol, 43%). .sup.1H NMR (CDCl.sub.3, 300 MHz): 7.40-7.30 (m, 1H),
7.00-6.91 (m, 2H), 6.30 (s, 1H), 5.20 (s, 2H), 4.37 (q, J=6.9 Hz,
2H), 2.37 (s, 3H), 2.31 (s, 3H), 1.35 (t, J=6.9 Hz, 3H); LRMS m/z
322.1 (M.sup.++1).
Step 1C
1-(2.6-Difluorobenzyl-2.6-dimethyl-3-bromo-5-ethoxycarbonyl-4-pyri-
done:
[0122]
1-(2,6-Difluorobenzyl)-2,6-dimethyl-3-ethoxycarbonyl-4-pyridone
(2.43 g, 7.6 mmol) was dissolved in glacial acetic acid (25 mL) and
treated with bromine (490 .mu.L, 9.5 mmol). The resulting mixture
was stirred at room temperature for 8 hours and poured into
ice/water. The product was extracted with dichloromethane, and the
extracts were washed with 10% aqueous NaHCO.sub.3, 5% aqueous
Na.sub.2S.sub.2O.sub.3 and brine. The organics were dried over
anhydrous MgSO.sub.4, filtered and evaporated in vacuo. The residue
was triturated with diethyl ether to give the product as a white
solid (1.32 g, 3.3 mmol, 43%). .sup.1H NMR (CDCl.sub.3, 300 MHz):
7.41-7.31 (m, 1H), 7.00-6.92 (m, 2H), 5.31 (s, 2H), 4.37 (q, J=6.9
Hz, 2H), 2.66 (s, 3H), 2.37 (s, 3H), 1.37 (t, J=6.9 Hz, 3H); LRMS
m/z 400.1 (M.sup.+-1), 402.1 (M.sup.++1).
EXAMPLE 2
SYNTHESIS OF
1-(2,6-DIFLUOROBENZYL)-2,6-DIMETHYL-3-(2-FLUORO-3-METHOXYPHENYL)-5-[2-(2--
PYRIDYL)ETHYLAMINOMETHYL]-4-PYRIDONE
[0123] ##STR12##
Step 2A
1-(2,6-Difluorobenzyl)-2.6-dimethyl-3-(2-fluoro-3-methoxyphenyl)-5-
-ethoxycarbonyl-4-pyridone
[0124]
1-(2,6-Difluorobenzyl)-2,6-dimethyl-3-bromo-5-ethoxycarbonyl-4-pyr-
idone (1.5 g, 3.7 mmol) was evacuated in a pressure vessel with
2-fluoro-3-methoxyphenyl boronic acid (764 mg, 4.5 mmol) and
tetrakis(triphenylphosphine) palladium (0) (428 mg, 370 .mu.mol). A
mixture of benzene/ethanol/1,2-dimethoxyethane (74 ml in a 10/1/11
ratio) and barium hydroxide (saturated aqueous, about 0.5 M) was
then added under vacuum, the vessel sealed under N.sub.2, and the
reaction stirred at 90.degree. C. for twelve hours, protected from
light. The organic layer was concentrated and purified using flash
silica chromatography (EtOAc/hexanes-1/4 to
MeOH/CH.sub.2Cl.sub.2-2/98), with the product eluting as the third
spot. The product was dried as an amber oil (1.12 g, 2.5 mmol,
68%). .sup.1H NMR (CDCl.sub.3, 300 MHz) .delta. 6.80-7.80 (m, 6H),
5.29 (s, 2H), 4.37-4.33 (m, 2H), 3.87 (s, 3H), 3.38 (s, 3H), 2.26
(s, 3H), 1.35 (t, 3H); LCMS=446 (MH.sup.+).
Step 2B
1-(2.6-Difluorobenzyl)-2,6-dimethyl-3-(2-fluoro-3-methoxyphenyl)-5-
-formyl-4-pyridone
[0125] Diisobutylaluminum hydride (3.75 mmol, 3.75 ml of a 1.0M
solution in hexanes) was added dropwise to a stirring solution of
1-(2,6-difluorobenzyl)-2,6-dimethyl-3-(2-fluoro-3-methoxyphenyl)-3-bromo--
5-ethoxycarbonyl-4-pyridone (1.12 g, 2.5 mmol) in THF/DCM (30 mL/12
mL), at -78.degree. C. under N.sub.2. A suspension was formed at
the end of the addition. After 1 hour, an additional aliquot of
DIBAL-H (2.5 mmol, 2.5 mL of a 1.0 M solution in hexanes) was
added. The reaction was stirred for 1 hour at -78.degree. C. MeOH
(50 mL) was carefully added to quench any unreacted DIBAL-H. The
solution was concentrated, then partitioned between EtOAc and 1 M
HCl. The organic layer was concentrated and purified using flash
silica gel chromatography (100% hexanes to EtOAc/hexanes 3/2). The
fourth spot which eluted was evaporated to give product as a cream
colored solid. .sup.1H NMR (CDCl.sub.3, 300 MHz) .delta. 10.55 (s,
1H), 6.80-7.75 (m, 6H), 5.36 (s, 2H), 3.90 (s, 3H), 2.79 (s, 3H),
2.27 (s, 3H); LCMS=402 (MH.sup.+).
Step 2C
1-(2.6-Difluorobenzyl)-2.6-dimethyl-3-(2-fluoro-3-methoxyphenyl)-5-
-[2-(2-pyridyl)ethylamino-4-pyridone
[0126] Sodium borohydride (30 mg, 794 .mu.mmol) was added to a
pre-mixed solution of
1-(2,6-difluorobenzyl)-2,6-dimethyl-3-(2-fluoro-3-methoxybenzyl)-5-formyl-
-4-pyridone (20 mg, 50 .mu.mol) and the amine (excess) in MeOH (1
mL). Upon addition of the reducing agent, the solution quickly
turned dark with the evolution of gas. Product formation was
instantaneous. The crude solution was concentrated, taken up in
acetonitrile, filtered and purified using a Gilson Prep-HPLC
system, with the product obtained as a TFA salt in 32% yield.
.sup.1H NMR (CDCl.sub.3, 300 MHz) .delta. 8.52 (d, 1H), 8.19 (t,
1H), 7.83 (d, 1H), 7.62 (t, 1H), 7.36 (m, 1H), 7.10 (t, 1H), 6.95
(m, 3H), 6.72 (t, 1H), 5.44 (s, 2H), 4.28 (s, 2H), 3.86 (s, 3H),
3.59 (m, 2H), 3.50 (m, 2H), 2.54 (s, 3H), 2.31 (s, 3H); MS=508
(MH.sup.+).
[0127] The following compounds were synthesized according to above
procedure. TABLE-US-00001 TABLE 1 ##STR13## No. R.sub.6 R.sub.1 MS
(MH.sup.+) 2-1 2-F-3- 2-PyridylCH.sub.2CH.sub.2 508 MeOPh 2-2
2-F-3- PhCH.sub.2 493 MeOPh 2-3 2-F-3- 2-MeOPhCH.sub.2 523 MeOPh
2-4 2-F-3- 2-MePhCH.sub.2 507 MeOPh 2-5 2-F-3- 2-PyridylCH.sub.2
494 MeOPh 2-6 3-MeOPh 2-PyridylCH.sub.2CH.sub.2 490 2-7 3-MeOPh
PhCH.sub.2 475 2-8 3-MeOPh 2-MeOPhCH.sub.2 505 2-9 3-MeOPh
2-MePhCH.sub.2 489 2-10 3-MeOPh ##STR14## 494 2-11 3-MeOPh
##STR15## 538 2-12 3-MeOPh Me.sub.2NCH.sub.2CH.sub.2 456 2-13
3-MeOPh (iPr).sub.2NCH.sub.2CH.sub.2 512 2-14 3-MeOPh
(HOCH.sub.2CH.sub.2).sub.2NCH.sub.2CH.sub.2 516 2-15 3-MeOPh
##STR16## 544 2-16 3-MeOPh ##STR17## 586 2-17 3-MeOPh ##STR18##
481.2 2-18 3-MeOPh ##STR19## 465.1 2-19 3-MeOPh ##STR20## 570.3
2-20 3-MeOPh ##STR21## 501.2 2-21 3-MeOPh PhCH(CH.sub.3) 489.2 2-22
3-MeOPh (CH.sub.3).sub.3CCH(CH.sub.3) 469.2 2-23 3-MeOPh
F.sub.3CCH.sub.2 467.1 2-24 3-MeOPh 2-MeO-PhCH.sub.2CH.sub.2 519.2
2-25 3-MeOPh 3,4-di-(OMe)PhCH.sub.2CH.sub.2 549.2 2-26 3-MeOPh
n-Heptyl 483.6 2-27 3-MeOPh 3-NO.sub.2PhCH.sub.2 520.5 2-28 3-MeOPh
CyclopropylCH.sub.2 439.2 2-29 3-MeOPh 4-ClPhCH(CH.sub.3) 523.1
2-30 3-MeOPh Et 413.5 2-31 3-MeOPh Allyl 425.5 2-32 3-MeOPh
Isopropyl 441.5 2-33 3-MeOPh 2-CF.sub.3-4-Cl-PhCH.sub.2 561.2 2-34
3-MeOPh 3,4-di-Cl-PhCH.sub.2 543.1 2-35 3-MeOPh
4-MeO-PhCH.sub.2CH.sub.2 519.2 2-36 3-MeOPh n-Propyl 427.1 2-37
3-MeOPh n-Butyl 441.5 2-38 3-MeOPh 3,4,5-tri-OMe-PhCH.sub.2 565.2
2-39 3-MeOPh CH.sub.3CH.sub.2OCH.sub.2CH.sub.2 457.5 2-40 3-MeOPh
CH.sub.3OCH.sub.2(CH.sub.3CH.sub.2)CH 471.5 2-41 2-F-3-
4-CH.sub.3-Cyclohexyl-CH.sub.2 499.2 MeOPh 2-42 2-F-3-
F.sub.3CCH.sub.2 485.1 MeOPh 2-43 2-F-3-
((CH.sub.3).sub.2CH).sub.2NCH.sub.2CH.sub.2 530.2 MeOPh 2-44 2-F-3-
Cyclopropyl-CH.sub.2 457.1 MeOPh 2-45 2-F-3- 4-Cl-PhCH.sub.2 527.1
MeOPh 2-46 Ph 4-CH.sub.3-Cyclohexyl-CH.sub.2 451.2 2-47 Ph
2-FurylCH.sub.2 435.1 2-48 Ph ##STR22## 540.2 2-49 Ph ##STR23##
471.2 2-50 Ph PhCH(CH.sub.3) 459.2 2-51 Ph
(CH.sub.3).sub.3CCH(CH.sub.3) 439.2 2-52 Ph CF.sub.3CH.sub.2 437.1
2-53 Ph 3,4-di-OMe-PhCH.sub.2CH.sub.2 519.2 2-54 Ph
(iPr).sub.2NCH.sub.2CH.sub.2 482.2 2-55 Ph
2-CF.sub.3-4-F-PhCH.sub.2 531.2 2-56 Ph 2-F-PhCH.sub.2CH.sub.2
477.2 2-57 Ph (S)-2-NaphthylCH(CH.sub.3) 509.2 2-58 Ph
4-OMe-PhCH.sub.2CH.sub.2 489.6 2-59 2-CH.sub.3- 2-FurylCH.sub.2
455.5 Thienyl 2-60 2-CH.sub.3- Thienyl ##STR24## 560.6 2-61
2-CH.sub.3- Thienyl ##STR25## 491.1 2-62 2-CH.sub.3- PhCH(CH.sub.3)
479.1 Thienyl 2-63 2-CH.sub.3- (CH.sub.3).sub.3CCH(CH.sub.3) 459.2
Thienyl 2-64 2-CH.sub.3- CF.sub.3CH.sub.2 457.5 Thienyl 2-65
2-CH.sub.3- 3,4-di-OMe-PhCH.sub.2CH.sub.2 539.7 Thienyl 2-66
2-CH.sub.3- 4-F-PhCH.sub.2CH.sub.2 497.6 Thienyl 2-67 2-CH.sub.3-
(CH.sub.3).sub.2CHCH.sub.2CH.sub.2CH(CH.sub.3) 473.6 Thienyl 2-68
2-CH.sub.3- (iPr).sub.2NCH.sub.2CH.sub.2 502.7 Thienyl 2-69
2-CH.sub.3- 4-Cl-PhCH(CH.sub.3) 514.1 Thienyl 2-70 2-CH.sub.3-
2-CF.sub.3-4-F-PhCH.sub.2 551.6 Thienyl 2-71 2-CH.sub.3-
2-F-PhCH.sub.2CH.sub.2 497.1 Thienyl 2-72 2-CH.sub.3- Thienyl
##STR26## 549.2 2-73 2-CH.sub.3- PhCH.sub.2 465.1 Thienyl 2-74
2-CH.sub.3- 3,4-di-Cl-PhCH.sub.2 535 Thienyl 2-75 2-CH.sub.3-
4-MeO-PhCH.sub.2CH.sub.2 509.2 Thienyl 2-76 2-CH.sub.3- n-Propyl
417.1 Thienyl 2-77 2-CH.sub.3- n-Butyl 431.1 Thienyl 2-78
2-CH.sub.3- CH.sub.3CH.sub.2OCH.sub.2CH.sub.2 447.6 Thienyl 2-79
2-CH.sub.3- CH.sub.3OCH.sub.2CH(CH.sub.2CH.sub.3) 461.6 Thienyl
2-80 2-CH.sub.3- 2-PyridylCH.sub.2CH.sub.2 480.6 Thienyl 2-81 Ph
Propyl 397.5 2-82 Ph CH.sub.3CH.sub.2OCH.sub.2CH.sub.2 427.5 2-83
Ph CH.sub.3OCH.sub.2CH(CH.sub.2CH.sub.3) 441.5 2-84 Ph
2-PyridylCH.sub.2CH.sub.2 460.5 2-85 2-F-3- 2-Pyridyl 480 MeOPh
2-86 2-F-3- 3-Pyridyl 480 MeOPh 2-87 2-F-3- 2-Pyridyl-6-Me 494
MeOPh 2-88 2-F-3- 2-Pyridyl-5-Me 494 MeOPh 2-89 2-F-3-
2-Pyridyl-3,5-diMe 508 MeOPh 2-90 2-F-3- 2-Pyridyl-4-Cl 514 MeOPh
2-91 2-F-3- 2-Pyridyl-4-Me 494 MeOPh 2-92 2-F-3- 2-Pyridyl-3-Me 494
MeOPh 2-93 2-F-3- MeOPh ##STR27## 530 2-94 2-F-3- MeOPh ##STR28##
530 2-95 2-F-3- 2-Pyridyl-5-F 498 MeOPh 2-96 2-F-3-
2-Pyrimidyl-4-Me 495 MeOPh 2-97 2-F-3- 2-Pyrimidyl-4,6-diMe 509
MeOPh 2-98 2-F-3- MeOPh ##STR29## 567 2-99 2-F-3- MeOPh ##STR30##
470 2-100 2-F-3- MeOPh ##STR31## 516 2-101 2-F-3- MeOPh ##STR32##
519 2-102 2-F-3- MeOPh ##STR33## 547 2-103 2-F-3- MeOPh ##STR34##
530 2-104 2-F-3- MeOPh ##STR35## 494 2-105 2-F-3- 2-ClPh 513 MeOPh
2-106 2-F-3- 2-MePh 493 MeOPh 2-107 2-F-3- 3-ClPh 513 MeOPh 2-108
2-F-3- 3-MeOPh 509 MeOPh 2-109 2-F-3- 4-MeOPh 509 MeOPh 2-110
2-F-3- 4-MePh 493 MeOPh
[0128] The following compounds were also synthesized according to
the above procedure. TABLE-US-00002 TABLE 2 ##STR36## No. R.sub.6
R.sub.1R.sub.2N MS (MH.sup.+) 2-111 2-F-3-MeOPh 1-Pyrrolidinyl
457.1 2-112 2-F-3-MeOPh 2,6-Dimethylmorpholinyl 501.2 2-113
2-CH.sub.3-Thienyl 1-Pyrrolidinyl 429.5 2-114 2-CH.sub.3-Thienyl
2,6-Dimethylmorpholinyl 473.2 2-115 Ph 1-Pyrrolidinyl 409.5
EXAMPLE 3
SYNTHESIS OF
1-(2,6-DIFLUOROBENZYL)-2,6-DIMETHYL-3-(2-FLUORO-3-METHOXYPHENYL)-5-[2-(2--
PYRIDYL)ETHYLAMINOMETHYL]-4-PYRIDONE
[0129] ##STR37##
Step 3A
1-(2.6-Difluorobenzyl)-2.6-dimethyl-3-(2-fluoro-3-methoxyphenyl)-5-
-[N-{2-(2-pyridyl)ethyl}-N-methylaminomethyl]-4-pyridone
[0130] Sodium triacetoxyborohydride (30 mg, 142 .mu.mol) was added
to a stirring solution of
1-(2,6-difluorobenzyl)-2,6-dimethyl-3-(2-fluoro-3-methoxybenzyl)-5-formyl-
-4-pyridone (20 mg, 50 .mu.mol) and 2-(2-methylaminoethyl)pyridine
(0.14 mL, 985 .mu.mol) in dichloromethane (1 mL). The solution
stirred overnight at r.t., MeOH (0.5 mL) was added and the solution
concentrated, taken up in acetonitrile, filtered and purified using
a Gilson Prep-HPLC system to give the product as a TFA salt in 34%
yield.
[0131] The following compounds are synthesized according to above
procedure. TABLE-US-00003 TABLE 3 ##STR38## No. R.sub.6
R.sub.1R.sub.2N-- MS (MH.sup.+) 3-1 2-F-3-MeOPh
2-PyCH.sub.2CH.sub.2NMe 522 3-2 2-F-3-MeOPh PhCH.sub.2NMe 507 3-3
2-F-3-MeOPh 2-PyCH.sub.2NMe 508 3-4 2-F-3-MeOPh 2-PyCH.sub.2NEt 522
3-5 2-F-3-MeOPh (2-PyCH.sub.2).sub.2N 585 3-6 3-MeOPh
2-PyCH.sub.2CH.sub.2NMe 504 3-7 3-MeOPh PhCH.sub.2NMe 489 3-8
2-F-3-MeOPh Et.sub.2N 516.2 3-9 2-CH.sub.3-Thienyl
PhCH.sub.2NCH.sub.3 479.6 3-10 2-CH.sub.3-Thienyl
2-PyCH.sub.2CH.sub.2NCH.sub.3 494.6 3-11 2-CH.sub.3-Thienyl
Et.sub.2NCH.sub.2CH.sub.2NCH.sub.3 488.7 3-12 Ph
PhCH.sub.2NCH.sub.3 459.6 3-13 Ph ButylNCH.sub.3 425.5 3-14 Ph
2-PyCH.sub.2CH.sub.2NCH.sub.3 474.2 3-15 Ph
Et.sub.2NCH.sub.2CH.sub.2NCH.sub.3 468.6
EXAMPLE 4
SYNTHESIS OF
1-(2,6-DIFLUOROBENZYL)-2,6-DIMETHYL-3-(2-FLUORO-3-METHOXYPHENYL)-5-[4-PRO-
PYLPIPERAZINYLMETHYL]-4-PYRIDONE
[0132] ##STR39##
Step 4A
1-(2,6-Difluorobenzyl)-2,6-dimethyl-3-(3-methoxyphenyl)-5-formyl-4-
-pyridone
[0133]
1-(2,6-Difluorobenzyl)-2,6-dimethyl-3-bromo-5-formyl-pyrid-4-one
(1.36 g, 3.8 mmol) stirred with 3-methoxyphenyl boronic acid (696
mg, 4.6 mmol) and tetrakis(triphenylphosphine)palladium(0) (439 mg,
380 .mu.mol) in benzene/ethanol/DME (76 mL, 10/1/11) and barium
hydroxide (30 mL, sat., aq.) under N.sub.2 at 90.degree. C.,
protected from light over 8 hours. The organic layer was then
concentrated and purified using flash silica chromatography (ethyl
acetate/hexane--1/9), giving the product as a cream-colored solid
(622 mg, 1.6 mmol, 42% yield). .sup.1H NMR (CDCl.sub.3, 300 MHz)
.delta. 10.55 (s, 1H), 7.41-7.31 (m, 2H), 7.00-6.74 (m, 5H), 5.35
(s, 2H), 3.81 (s, 3H), 2.79 (s, 3H), 2.25 (s, 3H); MS: 384
(MH.sup.+).
Step 4B
1-(2.6-Difluorobenzyl)-2,6-dimethyl-3-(3-methoxyphenyl)-5-[piperaz-
inylmethyl]-4-pyridone
[0134] Sodium triacetoxyborohydride (2.75 g, 13 mmol) was added to
a stirring solution of
1-(2,6-difluorobenzyl)-2,6-dimethyl-3-(3-methoxyphenyl)-5-formyl-4-pyrido-
ne (2.5 g, 6.5 mmol) and 1-boc-piperazine (1.33 g, 7.2 mmol) in
CH.sub.2Cl.sub.2 (45 mL) at r.t. The solution was quenched with 1M
KOH (5 mL) and the organic layer purified using flash silica
chromatography (1% MeOH in CH.sub.2Cl.sub.2) to give the boc
protected product as a dark brown oil. The resulting oil was
treated with trifluoroacetic acid (5 mL, 6.5 mmol) in
CH.sub.2Cl.sub.2 (20 mL) for one hour at r.t. The crude solution
was concentrated and purified using flash silica chromatography (2%
MeOH in CH.sub.2Cl.sub.2 and dried to give the product as a white
foam (1.9 g, 3.4 mmol, 52% yield). .sup.1H NMR (CDCl.sub.3, 300
MHz) .delta. 7.38-7.28 (m, 2H), 6.97-6.74 (m, 5H), 5.30 (s, 2, H),
3.80 (s, 3H), 3.49 (s, 2H), 3.20-2.80 (m, 8H), 2.52 (m, 3H) 2.19
(s, 3H). MS: 568 (MH.sup.+).
Step 4C
1-(2.6-Difluorobenzyl)-2.6-dimethyl-3-(3-methoxyphenyl)-5-[4-(2-et-
hoxycarbonylcyclopropanemethyl)piperazinylmethyl]-4-pyridone
[0135] Sodium triacetoxyborohydride (30 mg, 141 .mu.mol) was added
to a stirring solution of
1-(2,6-difluorobenzyl)-2,6-dimethyl-3-(3-methoxyphenyl)-5-[piperazinylmet-
hyl]-4-pyridone (50 mg, 88 .mu.mol) with
ethyl-2-formyl-1-cyclopropane carboxylate (0.05 mL, 669 .mu.mol) in
CH.sub.2Cl.sub.2 (1 mL). The mixture stirred at r.t. for five hours
before being filtered and purified using the Gilson Prep-HPLC
system, giving the product as a TFA salt (13.4 mg, 19 .mu.mol, 22%
yield). .sup.1H NMR (CDCl.sub.3, 300 MHz) .delta. 7.40-7.32 (m,
2H), 7.00-6.69 (m, 5H), 5.42 (s, 2H), 4.37 (s, 2H), 4.194.07 (m,
2H), 3.80 (s, 3H), 3.68-3.48 (m, 8H), 3.00-2.88 (m, 1H), 2.58 (s,
3H), 2.28 (m, 3H), 1.67-1.54 (m, 3H), 1.36-1.22 (m, 5H). MS: 580
(MH.sup.+). TABLE-US-00004 TABLE 4 ##STR40## No. R MS (MH.sup.+)
4-1 H 454 4-2 EtOCO(cPr)CH.sub.2 580 4-3 5-MeFuranCH.sub.2 548 4-4
3,4-MeOPhCH.sub.2 604 4-5 1-NaphCH.sub.2 594 4-6 iBu 510 4-7 Pr 496
4-8 c-PrCH.sub.2 508
EXAMPLE 5
SYNTHESIS OF
1-(2,6-DIFLUOROBENZYL)-2,6-DIMETHYL-3-(2-FLUORO-3-METHOXYPHENYL)-5-[N-{2--
(2-PYRIDYL)ETHYL}-N-METHYLAMINOMETHYL]4-PYRIDONE
[0136] ##STR41##
Step 5A
1-(2,6-Difluorobenzyl)-2,6-dimethyl-3-bromo-5-formyl-4-pyridone
[0137] DIBAL (5.3 mmol, 5.3 mL of a 1.0 M solution in hexanes) was
added dropwise via syringe to a stirred solution of
1-(2,6-difiuorobenzyl)-2,6-dimethyl-3-bromo-5-ethoxycarbonyl-4-pyridone
(1.40 g, 3.5 mmol) in THFIDCM (60 mL/20 mL), at -78.degree. C.
under N.sub.2. A suspension was formed at the end of the addition.
After 1 hour, an additional aliquot of DIBAL (3.0 mmol, 3.0 mL of a
1.0 M solution in hexanes) was added. The reaction was deemed
complete after 1 hour at -78.degree. C. MeOH (5 mL) was carefully
added to quench any unreacted DIBAL and the mixture was partitioned
between EtOAc and 1 M HCl. The organic layer was separated and
washed with brine, dried over anhydrous MgSO.sub.4 and filtered.
Upon removal of the solvents in vacuum, the crude solid residue was
triturated with Et.sub.2O. The product was obtained as a beige
solid (0.80 g, 2.25 mmol, 64%). .sup.1H NMR (CDCl.sub.3, 3000 MHZ):
10.55 (s, 1H), 7.40-7.33 (m, 1H), 7.00-6.93 (m, 2H), 5.38 (s, 2H),
2.78 (s, 3H), 2.69 (s, 3H); LPMS m/z 356.0 (M.sup.+-1), 358.0
(M.sup.++1).
Step 5B
1-(2,6-Difluorobenzyl)-2,6-dimethyl-3-bromo-5-(2-methoxyethenyl)4--
pyridone
[0138] KHMDS (2.4 mmol, 4.8 mL of a 0.5 M solution in toluene) was
added dropwise to a vigorously stirred suspension of
(methoxymethyl)triphenylphosphonium chloride (771 mg, 2.25 mmol) in
anhydrous THF (8 mL), at -78.degree. C. under N.sub.2. The
resulting orange suspension was stirred at -78.degree. C. for 45
min. Then, a solution of
1-(2,6-difluorobenzyl)-2,6-dimethyl-3-bromo-5-formyl-4-pyridone
(534 mg, 1.50 mmol) in THF (8 mL) was added dropwise and the
reaction was allowed to warm slowly to room temperature. After 1 h
at room temperature, the reaction was quenched with an aqueous
saturated solution of NaHCO.sub.3. The organics were extracted with
EtOAc (50 mL), the organic layer was separated, washed with
H.sub.2O and brine, dried over MgSO.sub.4, filtered and evaporated
under vacuum. The crude residue was utilized in the next step. LRMS
m/z 384.0 (M.sup.+-1), 386.0 (M.sup.++1).
Step
5C.sub.1-(2.6-Difluorobenzyl)-2.6-dimethyl-3-bromo-5-(carbonylmethyl)-
4-pyridone:
[0139] The crude product above was dissolved in THF/H.sub.2O (20
mL, 1:1 v/v) and to that solution TFA (5 mL) was added. The mixture
was heated at reflux for 19 h. After cooling down, the reaction was
extracted with CH.sub.2Cl.sub.2 (50 mL), washed with sat.
NaHCO.sub.3 and brine. After drying over MgSO.sub.4, the solvent
was removed under vacuum and the residue was chromatographed on
silica-gel, eluting with EtOAc. The product was obtained as a pale
yellow oil (431 mg, 1.16 mmol, 78% over 2 steps). .sup.1H NMR
(CDCl.sub.3, 300 MHz) .delta. 9.56 (t, J=1.8 Hz, 1H), 7.38-7.33 (m,
1H), 6.98-6.93 (m, 2H), 5.37 (s, 2H), 3.83 (d, J=1.8 Hz, 2H), 2.68
(s, 3H), 2.33 (s, 3H); LRMS m/z 247.1 [M.sup.+-(Br, CH.sub.2CHO)],
370.0 (M.sup.+-1), 372.0 (M.sup.++1).
Step 5D
1-(2.6-Difluorobenzyl)-2,6-dimethyl-3-bromo-5-[N-{2-(2-pyridyl)eth-
yl}-N-methylaminomethyl]-4-pyridone
[0140] (2-Methylaminoethyl)pyridine (680 mg, 5.00 mmol) was added
to a stirred solution of
1-(2,6-difluorobenzyl)-2,6-dimethyl-3-bromo-5-(carbonylmethyl)4-pyridone
(430 mg, 1.16 mmol) in CH.sub.2Cl.sub.2 (15 mL). After 30 min.,
NaBH(OAc).sub.3 (1.23 g, 5.80 mmol) was added in small portions,
and the resulting mixture was stirred at room temperature for 1 h.
The reaction mixture was diluted with H.sub.2O (15 mL) and the
organic layer was separated and washed with sat. NaHCO.sub.3 and
brine. The organics were dried over MgSO.sub.4, filtered and
concentrated in vacuo. The crude was used in the next step without
any further purification. Crude yield 426 mg (75%). LRMS m/z 490.0
(M.sup.+-1), 492.0 (M.sup.++1).
Step 5E
1-(2,6-Difluorobenzyl)-2.6-dimethyl-3-(2-fluoro-3-methoxyphenyl)-5-
-[N-{2-(2-pyridyl)ethyl}-N-methylaminomethyl]4-pyridone
[0141] The above crude bromide (98 mg, 0.20 mmol) and
3-methoxyphenyl boronic acid (36 mg, 0.24 mmol) were dissolved in
DME/benzene/EtOH (11:10:1, 5 mL). To that, saturated aqueous
Ba(OH).sub.2 (1.4 mL) was added and the mixture was degassed under
N.sub.2 for 30 minutes. Pd[Ph.sub.3P].sub.4 (23 mg, 0.02 mmol) was
then added and the sealed reaction vessel was stirred at 80.degree.
C. for 18 h. The reaction mixture was cooled down, extracted with
EtOAc, washed with H.sub.2O and brine, dried over MgSO.sub.4,
filtered and evaporated. The residue was purified by preparative
TLC plate (0.5 mm, 20.times.20 cm), eluting with a
CHCl.sub.3/MeOH/NH.sub.4OH (88.5:11.0:0.5) mixture to give product
(15 mg, 0.03 mmol, 15% yield). .sup.1H NMR (CDCl.sub.3, 300 MHz)
.delta. 8.50 (d, J=4.2 Hz, 1H), 7.57 (dt, J=1.8 Hz, J.sub.2=7.5 Hz,
1H), 7.36-7.27 (m, 2H), 7.20 (d, J=7.5 Hz, 1H), 7.11-7.07 (m, 1H),
6.97-6.92 (m, 2H), 6.86-6.73 (m, 3H), 5.30 (s, 2H), 3.80 (s, 3H),
3.03-2.97 (m, 2H), 2.92-2.83 (m, 4H), 2.62-2.56 (m, 2H), 2.44 (s,
3H), 2.40 (s, 3H), 2.21 (s, 3H); LRMS m/z 518.3 (M.sup.++1).
[0142] The following compounds are synthesized according to above
procedure. TABLE-US-00005 TABLE 5 ##STR42## No. R.sub.6
R.sub.1R.sub.2N MS (MH.sup.+) 5-1 2-F-3-MeOPh
2-PyCH.sub.2CH.sub.2NMe 536 5-2 3,4-CH.sub.2O.sub.2Ph
2-PyCH.sub.2CH.sub.2NMe 532 5-3 4-iPrPh 2-PyCH.sub.2CH.sub.2NMe 530
5-4 3-MeOPh 2-PyCH.sub.2CH.sub.2NMe 518
EXAMPLE 6
SYNTHESIS OF
1-(2,6-DIFLUOROBENZYL)-2-METHYL-3-[(2R)-AMINO-2-PHENYLETHOXYL]-5-ARYLPYRI-
DIN-4-ONE
[0143] ##STR43##
Step 6A
2-Methyl-3-[(2R)-Boc-amino-2-phenylethoxy]-4H-pyran-4-one
[0144] Diethyl azodicarboxylate (2.36 mL, 15.0 mmol) was added
dropwise to a stirred solution of Maltol (1.26 g, 10.0 mmol),
triphenylphosphine (3.93 g, 15.0 mmol) and (R)-N-boc-phenylglycinol
(2.37 g, 10.0 mmol) in THF (100 mL). The initial yellow color faded
quickly, and the resulting colorless solution was stirred under
N.sub.2 for 15 h at room temperature. The solvent was removed in
vacuo and the residue was purified on silica gel, eluting with a
2:1 v/v mixture of hexanes/EtOAc. The title product was obtained as
a yellow oil, which was contaminated with some triphenylphosphine
oxide. This mixture was carried onto the next step without any
further purification. Yield=2.35 g. LRMS m/z 246 (M.sup.+-boc+1),
229 (246-NH.sub.3).
Step 6B
1-(2,6-Difluorobenzyl)-2-methyl-3-[(2R)-Boc-amino-2-phenylethoxy]--
pyridin-4-one
[0145] Crude
2-methyl-3-[(2R)-Boc-amino-2-phenylethoxy]-4H-pyran-4-one (1.00 g,
.about.2.90 mmol), 2,6-difluorobenzyl amine (700 .mu.L, 5.8 mmol)
and ethanol (3 mL) were combined and heated to 120.degree. C. in a
pressure vessel. After 16 h, the volatiles were removed in vacuo
and the residue purified by column chromatography, eluting with
EtOAc. The product was isolated as a white foam (204 mg, 0.43 mmol,
15%, assuming pure starting material). .sup.1H NMR (CDCl.sub.3-300
MHz) .delta. 7.42-7.19 (m, 7H); 6.98 (t, J=8.1 Hz, 2H); 6.43 (d,
J=7.8 Hz, 1H); 5.06 (s, 2H); 4.87-4.83 (m, 1H); 4.06 (dd, J=9.9 Hz,
J.sub.2=4.2 Hz, 1H); 3.93 (dd, J=9.9 Hz, J.sub.2=6.8 Hz, 1H); 2.34
(s, 3H); 1.42 (s, 9H); LRMS m/z 471.1 (M.sup.++1), 371.0
(M.sup.+-boc+1).
Step 6C
1-(2,6-Difluorobenzyl)-2-methyl-3-[(2R)-Boc-amino-2-phenylethoxy]--
5-bromopyridin-4-one
[0146] N-Bromosuccinimide (79 mg, 0.44 mmol) was added to a stirred
solution of
1-(2,6-difluorobenzyl)-2-methyl-3-[(2R)-Boc-amino-2-phenylethoxy]-5-bromo-
pyridin-4-one (173 mg, 0.37 mmol) in THF (2 mL), under an
atmosphere of N.sub.2. The resulting mixture was refluxed for 2 h
and the mixture was cooled to room temperature and partitioned
between EtOAc and H.sub.2O. The organic layer was separated, washed
with aqueous NaHCO.sub.3 and brine, dried over anhydrous
MgSO.sub.4, filtered and concentrated in vacuo. The product was
obtained as a yellow foam (185 mg, 0.34 mmol, 92%). .sup.1H NMR
(CDCl.sub.3-300 MHz) .delta. 7.76 (s, 1H); 7.47-7.19 (m, 6H); 7.00
(t, J=8.1 Hz, 2H); 5.09 (s, 2H); 4.90-4.87 (m, 1H); 4.09-4.00 (m,
2H); 2.34 (s, 3H); 1.41 (s, 9H); LRMS m/z 551.1 (M++3), 549.1
(M.sup.++1), 451.0 (M.sup.+-boc+3), 449.0 (M.sup.+-boc+1).
Step 6D
1-(2.6-Difluorobenzyl)-2-methyl-3-[(2R)-Boc-amino-2-phenylethoxy]--
5-arylpyridin-4-one (General procedure for Suzuki counlings)
[0147] The aryl boronic acid (1.3 mmol) and aqueous saturated
Ba(OH).sub.2 solution (7.1 mL) were added to a solution of
1-(2,6-difluorobenzyl)-2-methyl-3-[(2R)-Boc-amino-2-phenylethoxy]-5-bromo-
pyridin-4-one (1.0 mmol) in a solvent mixture consisting of
dimethoxyethane, benzene and ethanol, in a 50:45.5:4.5 ratio (20
mL), respectively. The resulting mixture was degassed by bubbling
N.sub.2 for about 20 minutes. Pd[PPh.sub.3].sub.4 (0.1 mmol) was
then added, the vessel was sealed and immersed in an oil bath at
90.degree. C., with stirring. The reaction was monitored by TLC and
LC/MS. When the starting material was consumed (typically 5-10
hours), the reaction was cooled and partitioned between EtOAc and
water. The organic layer was washed with brine, dried over
MgSO.sub.4 and filtered. The filtrates were concentrated in vacuum
and the residue was purified by column chromatography on silica
gel. Yields typically range from 65-95%.
Step 6E
1-(2.6-Difluorobenzyl)-2-methyl-3-[(2R)-amino-2-phenylethoxy]-5-ar-
ylpyridin-4-one (General Procedure for N-boc deprotections)
[0148] Trifluoroacetic acid (2 mL) was added to a solution of
1-(2,6-difluorobenzyl)-2-methyl-3-[(2R)-Boc-amino-2-phenylethoxy]-5-arylp-
yridin-4-one (0.09 mmol) in CH.sub.2Cl.sub.2 (2 mL) and the
resulting mixture was stirred at room temperature for 2 h. TLC and
LC/MS analysis showed that all starting material had been consumed.
The volatiles were removed in vacuo and the residues were either
purified directly by preparative HPLC or neutralized with NH.sub.3
in MeOH and then purified by preparative TLC, eluting with a
88:11:1 v/v mixture of CHCl.sub.3/MeOH/NH.sub.4OH. TABLE-US-00006
TABLE 6 ##STR44## MS No. R.sub.6 .sup.1H NMR(CDCl.sub.3, 300 MHz)
(MH.sup.+) 6-1 3-MeOPh 7.55(s, 1 H); 7.44-7.22 (m, 8H); 477 7.11
(dd, J.sub.1 = 1.0 Hz, J.sub.2 = 7.7 Hz, 1 H); 6.99 (t, J = 8.1 Hz,
2 H); 6.86 (dd, J.sub.1 = 1.7 Hz, J.sub.2 = 8.3 Hz, 1 H); 5.12 (s,
2 H); 4.47-4.43 (m, 1 H); 4.19 (dd, J.sub.1 = 4.1 Hz, J.sub.2 = 9.5
Hz, 1 H); 4.00 (dd, J.sub.1 = 9.0 Hz, J.sub.2 = 9.5 Hz, 1 H); 3.83
(s, 3 H); 2.37 (s, 3 H) 6-2 3,4-CH.sub.2O.sub.2Ph 7.49 (s, 1 H);
7.44-7.22 (m, 6 H), 491 7.16 (d, J = 1.8 Hz, 1 H), 7.01-6.96 (m, 3
H); 6.83 (d, J = 8.1 Hz, 1 H); 5.95 (s, 2 H); 5.11 (s, 2 H); 4.46-
4.42 (m, 1 H); 4.17 (dd, J.sub.1 = 4.0 Hz, J.sub.2 = 9.3 Hz, 1 H);
3.99 (dd, J.sub.1 = 8.7 Hz, J.sub.2 = 9.3 Hz, 1 H); 2.36 (s, 3 H)
6-3 2-F-3-MeOPh 495
EXAMPLE 7
Synthesis of
1-(2,6-difluorobenzyl)-2,6-dimethyl-3-[N-(2-aminoethyl)aminomethyl]-5-(2--
fluoro-3-methoxy)pyridin-4-one
[0149] ##STR45##
Step 7A
1-(2,6-Difluorobenzyl)-2,6-dimethyl-3-(2-fluoro-3-methoxyphenyl)-5-
-(2-aminoethyl)methylamino-4-pyridone
[0150] Butyl(tert)-N-(2-aminoethyl)carbamate (530 mg, 3.3 mmol) was
added to a stirring solution of
1-(2,6-Difluorobenzyl)-2,6-dimethyl-3-(2-fluoro-3-methoxyphenyl)-5-formyl-
-4-pyridone (870 mg, 2.2 mmol) and magnesium sulfate (145 mg, 1.2
mmol) in methanol (29 mL) at room temperature. The resulting
mixture stirred 1 hour before addition of sodium borohydride (145
mg, 3.8 mmol). The crude reaction was concentrated in vacuo and
purified by Flash silica chromatography, and dried as an amber oil.
Yield 460 mg (843 .mu.mol, 38%); LCMS m/z 546 (M.sup.++1).
[0151] Trifluoroacetic acid (10 mL, 130 mmol) was added to the
boc-protected diaminoethylpyridone (460 mg, 843 .mu.mol) in
dichloromethane (20 mL) and stirred for 8 h at r.t. The material
was dried in vacuo as an oil, as TFA salt. Yield 470 mg (840
.mu.mol, 99%). .sup.1H NMR (CDCl.sub.3, 300 MHz) .delta. 7.37-7.29
(m, 1H), 7.13-7.09 (m, 1H), 6.97-6.92 (m, 3H), 6.83-6.79 (m, 1H),
5.34 (s, 2H), 3.89 (s, 3H), 3.84 (s, 2H), 3.39 (m, 2H), 3.08 (m,
2H), 2.50 (s, 3H), 2.25 (s, 3H); LCMS m/z 446 (M.sup.++1).
EXAMPLE 9
Synthesis of
1-(2,6-difluorobenzyl)-2,6-dimethyl-3-[N-(2-phenyl-2-imidiazoline)methyl]-
-5-(2-fluror-3-methoxyphenyl)-4-pyridinone
[0152] ##STR46##
[0153] Ethyl alcohol (1 mL, 17.7 mmol) was added to
1-(2,6-difluorobenzyl)-2,6-dimethyl-3-[N-(2-aminoethyl)aminomethyl]-5-(2--
fluoro-3-methoxy)-4-pyridone TFA salt (95 mg, 170 .mu.mol) with
methyl benzimidate hydrochloride (35 mg, 204 .mu.mol), sealed in a
glass pressure vessel and stirred for 2 h at 75.degree. C. The
crude material was purified using Prep-HPLC-MS, and dried in vacuo
as an oil (TFA salt). Yield 30.7 mg (48 .mu.mol, 28%). .sup.1H NMR
(CDCl.sub.3, 300 MHz) .delta. 7.66-7.60 (m, 3H), 7.54-7.49 (m, 2H),
7.42-7.32 (m, 1H), 7.14-7.08 (m, 1H), 7.03-6.93 (m, 3H), 6.70-6.65
(m, 1H), 5.44 (s, 2H), 4.74 (s, 2H), 3.91(m, 3H), 3.88 (m, 4H),
2.37 (s, 3H), 2.30 (s, 3H); LCMS m/z 532 (M.sup.++1).
EXAMPLE 10
Synthesis of
1-(2,6-difluorobenzyl)-2-methyl-3-[N-(quanyl)-N'-phenylaminomethyl]-5-(2--
fluoro-3-methoxyphenyl)pyridin-4-one
[0154] ##STR47##
[0155] N,N'-bis-boc-1-guanylpyrazole (195 mg, 627 .mu.mol) was
added to
1-(2,6-difluorobenzyl)-2-methyl-3-(N-phenylaminomethyl)-5-(2-fluoro-3-met-
hoxyphenyl)pyridin-4-one (150 mg, 313 .mu.mol) in 1,4-dioxane (6
mL), sealed in a glass pressure vessel and stirred for 8 h at
100.degree. C. The crude material was purified using Flash silica
chromatography, and dried in vacuo as a clear oil. Yield 50 mg (69
mmol, 22%).
[0156] Trifluoroacetic acid (2 mL, 26 mmol) was added to the
boc-protected guanylpyridone (50 mg, 69 .mu.mol) in dichloromethane
(5 mL) and stirred for 2 h at r.t. The crude material was purified
using Flash silica chromatography, and dried in vacuo as a clear
yellow oil. Yield 2 mg (3.8 .mu.mol, 5.5%). .sup.1H NMR
(CDCl.sub.3, 300 MHz) .delta. 7.30-7.20 (m, 4H), 7.05-6.82 (m, 6H),
6.60-6.50 (m, 1H), 5.45 (s, 2H), 5.40-5.15 (m, 2H), 3.85 (s, 3H),
2.40 (s, 3H), 2.15 (s, 3H); LCMS m/z 521 (M.sup.++1).
[0157] It will be appreciated that, although specific embodiments
of the invention have been described herein for purposes of
illustration, various modifications may be made without departing
from the spirit and scope of the invention. Accordingly, the
invention is not limited except as by the appended claims.
[0158] All of the above U.S. patents, U.S. patent application
publications, U.S. patent applications, foreign patents, foreign
patent applications and non-patent publications referred to int his
specification and/or listed in the Application Data Sheet are
incorporated herein by reference, in their entirety.
* * * * *