U.S. patent application number 10/705723 was filed with the patent office on 2004-08-19 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, Guo, Zhiqiang, Regan, Collin, Tucci, Fabio, Wu, Dongpei.
Application Number | 20040162290 10/705723 |
Document ID | / |
Family ID | 23200637 |
Filed Date | 2004-08-19 |
United States Patent
Application |
20040162290 |
Kind Code |
A1 |
Guo, Zhiqiang ; et
al. |
August 19, 2004 |
Gonadotropin-releasing hormone receptor antagonists and methods
relating thereto
Abstract
GnRH 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: 1 wherein R.sub.1, R.sub.2, R.sub.3a, R.sub.3b, R.sub.4,
R.sub.5, 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: |
Guo, Zhiqiang; (San Diego,
CA) ; Wu, Dongpei; (San Diego, CA) ; Chen,
Chen; (San Diego, CA) ; Tucci, Fabio; (San
Diego, CA) ; Regan, Collin; (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
|
Family ID: |
23200637 |
Appl. No.: |
10/705723 |
Filed: |
November 10, 2003 |
Related U.S. Patent Documents
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Application
Number |
Filing Date |
Patent Number |
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10705723 |
Nov 10, 2003 |
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10211955 |
Aug 2, 2002 |
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6677340 |
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60310018 |
Aug 2, 2001 |
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Current U.S.
Class: |
514/241 ;
544/216 |
Current CPC
Class: |
A61P 19/08 20180101;
A61P 3/14 20180101; A61P 15/10 20180101; C07D 409/06 20130101; A61P
5/24 20180101; A61P 5/30 20180101; A61P 1/00 20180101; C07D 417/10
20130101; C07D 417/14 20130101; A61P 13/08 20180101; A61P 5/32
20180101; A61P 11/00 20180101; A61P 37/02 20180101; A61P 35/00
20180101; A61P 15/08 20180101; A61P 5/34 20180101; C07D 401/12
20130101; C07D 417/04 20130101; A61P 9/12 20180101; A61P 43/00
20180101; A61P 5/36 20180101; A61P 15/18 20180101; C07D 403/06
20130101; C07D 403/12 20130101; A61P 5/26 20180101; A61P 5/06
20180101; A61P 15/00 20180101; C07D 251/34 20130101; A61P 25/20
20180101 |
Class at
Publication: |
514/241 ;
544/216 |
International
Class: |
A61K 031/53 |
Goverment Interests
[0002] Partial funding of the work described herein was provided by
the U.S. Government under Grant No. R43-HD38625 provided by the
National Institutes of Health. The U.S. Government may have certain
rights in this invention.
Claims
We claim:
1. A compound having the following structure: 227or a stereoisomer,
prodrug or pharmaceutically acceptable salt thereof, n is 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 arylalkyl, substituted arylalkyl,
heteroarylalkyl or substituted heteroarylalkyl; R.sub.5 is aryl,
substituted aryl, heteroaryl or substituted heteroaryl; R.sub.8 is
independently hydrogen, alkyl, substituted alkyl, aryl, substituted
aryl, arylalkyl, substituted arylalkyl, heterocycle, substituted
heterocycle, heterocyclealkyl or substituted heterocyclealkyl;
R.sub.9 is independently hydrogen, alkyl, substituted alkyl, aryl,
substituted aryl, arylalkyl, substituted arylalkyl, heterocycle,
substituted heterocycle, heterocyclealkyl or substituted
heterocyclealkyl; R.sub.10 and R.sub.11, are the same or different
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.
2. The compound of claim 1 wherein R.sub.4 is substituted arylalkyl
or substituted heteroarylalkyl.
3. The compound of claim 1 wherein R.sub.5 is substituted aryl or
substituted heteroaryl.
4. The compound of claim 1 wherein n is 2 or 3.
5. The compound of claim 4 wherein R.sub.4 is substituted arylalkyl
or substituted heteroarylalkyl.
6. A pharmaceutical composition comprising a compound of claim 1
and a pharmaceutically acceptable carrier or diluent.
7. A method for antagonizing gonadotropin-releasing hormone in a
subject in need thereof, comprising administering to the subject an
effective amount of a compound of claim 1.
8. A method for treating a sex-hormone related condition of a
subject in need thereof, comprising administering to the subject an
effective amount of the pharmaceutical composition of claim 6.
9. The method of claim 8 wherein the sex-hormone related condition
is cancer, benign prostatic hypertrophy or myoma of the uterus.
10. The method of claim 9 wherein the cancer is prostatic cancer,
uterine cancer, breast cancer or pituitary gonadotroph
adenomas.
11. The method of claim 8 wherein the sex-hormone related condition
is endometriosis, polycystic ovarian disease, uterine fibroids or
precocious puberty.
12. A method for preventing pregnancy of a subject in need thereof,
comprising administering an effective amount of the pharmaceutical
composition of claim 6.
13. 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 the pharmaceutical composition
of claim 6.
Description
CROSS-REFERENCES TO RELATED APPLICATIONS
[0001] This application is a continuation of U.S. application Ser.
No. 10/211,955 filed Aug. 2, 2002, which claims the benefit of U.S.
Provisional Patent Application No. 60/310,018 filed Aug. 2, 2001,
which applications are incorporated herein by reference in their
entirety.
BACKGROUND OF THE INVENTION
[0003] 1. Technical Field
[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-L- eu-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): 2
[0012] including stereoisomers, prodrugs and pharmaceutically
acceptable salts thereof, wherein R.sub.1, R.sub.2, R.sub.3a,
R.sub.3b, R.sub.4, R.sub.5 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): 3
[0017] including stereoisomers, prodrugs and pharmaceutically
acceptable salts thereof, wherein:
[0018] n is 2, 3 or 4;
[0019] 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);
[0020] or R.sub.1 and R.sub.2 taken together with the nitrogen atom
to which they are attached form a heterocycle or substituted
heterocycle;
[0021] 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;
[0022] 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;
[0023] 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;
[0024] R.sub.4 is arylalkyl, substituted arylalkyl, heteroarylalkyl
or substituted heteroarylalkyl;
[0025] R.sub.5 is aryl, substituted aryl, heteroaryl or substituted
heteroaryl;
[0026] R.sub.8 is independently hydrogen, alkyl, substituted alkyl,
aryl, substituted aryl, arylalkyl, substituted arylalkyl,
heterocycle, substituted heterocycle, heterocyclealkyl or
substituted heterocyclealkyl;
[0027] R.sub.9 is independently hydrogen, alkyl, substituted alkyl,
aryl, substituted aryl, arylalkyl, substituted arylalkyl,
heterocycle, substituted heterocycle, heterocyclealkyl or
substituted heterocyclealkyl;
[0028] R.sub.10 and R.sub.11, are the same or different
independently hydrogen, alkyl, substituted alkyl, aryl, substituted
aryl, arylalkyl, substituted arylalkyl, heterocycle, substituted
heterocycle, heterocyclealkyl or substituted heterocyclealkyl;
and
[0029] R.sub.12 is hydrogen, alkyl, or substituted alkyl.
[0030] As used herein, the above terms have the following
meaning:
[0031] "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.
[0032] "Aryl" means an aromatic carbocyclic moiety such as phenyl
or naphthyl.
[0033] "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.
[0034] "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.
[0035] "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.
[0036] "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.
[0037] "Heterocyclealkyl" means an alkyl having at least one alkyl
hydrogen atom replaced with a heterocycle, such as
--CH.sub.2morpholinyl, and the like.
[0038] "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.
[0039] 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.
[0040] "Halogen" means fluoro, chloro, bromo and iodo.
[0041] "Haloalkyl" means an alkyl having at least one hydrogen atom
replaced with halogen, such as trifluoromethyl and the like.
[0042] "Alkoxy" means an alkyl moiety attached through an oxygen
bridge (i.e., --O-alkyl) such as methoxy, ethoxy, and the like.
[0043] "Alkylthio" means an alkyl moiety attached through a sulfur
bridge (i.e., --S-alkyl) such as methylthio, ethylthio, and the
like.
[0044] "Alkylsulfonyl" means an alkyl moiety attached through a
sulfonyl bridge (i.e., --SO.sub.2-alkyl) such as methylsulfonyl,
ethylsulfonyl, and the like.
[0045] "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.
[0046] With regard to the
"R.sub.1R.sub.2N(CR.sub.3aR.sub.3b).sub.n--" moiety of structure
(I), n may be 2, 3 or 4. Accordingly, this moiety may be
represented by the following structure (i) when n is 2, structure
(ii) when n is 3, and structure (iii) when n is 4: 4
[0047] 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).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. 567
[0049] Representative compounds of this invention include those
listed below. With regard to nomenclature, the compounds of
structure (I) may generally be referred to as substituted
isocyanurates or, more specifically, as substituted
[1,3,5]triazinane-2,4,6-triones:
[0050]
1-(2-Amino-2-phenyl-ethyl)-3-(2,6-difluoro-benzyl)-5-(2-fluoro-3-me-
thoxy-phenyl)-[1,3,5]triazinane-2,4,6-trione;
[0051]
1-(2-Amino-2-phenyl-ethyl)-3-(2,6-dichloro-benzyl)-5-(2-fluoro-3-me-
thoxy-phenyl)-[1,3,5]triazinane-2,4,6-trione;
[0052]
1-(2-Amino-2-phenyl-ethyl)-3-(2-fluoro-6-chloro-benzyl)-5-(2-fluoro-
-3-methoxy-phenyl)-[1,3,5]triazinane-2,4,6-trione;
[0053]
1-(2-Amino-2-phenyl-ethyl)-3-(2-fluoro-6-trifluoromethyl-benzyl)-5--
(2-fluoro-3-methoxy-phenyl)-[1,3,5]triazinane-2,4,6-trione;
[0054]
1-(2-Amino-2-phenyl-ethyl)-3-(2-fluoro-6-methylsulfonyl-benzyl)-5-(-
2-fluoro-3-methoxy-phenyl)-[1,3,5]triazinane-2,4,6-trione;
[0055]
1-(2-Amino-2-phenyl-ethyl)-3-(2-fluoro-benzyl)-5-(2-fluoro-3-methox-
y-phenyl)-[1,3,5]triazinane-2,4,6-trione;
[0056]
1-(2-Amino-2-phenyl-ethyl)-3-(2-chloro-benzyl)-5-(2-fluoro-3-methox-
y-phenyl)-[1,3,5]triazinane-2,4,6-trione;
[0057]
1-(2-Amino-2-phenyl-ethyl)-3-(2-trifluoromethyl-benzyl)-5-(2-fluoro-
-3-methoxy-phenyl)-[1,3,5]triazinane-2,4,6-trione;
[0058] 1-(2-Amino-2-phenyl-ethyl)-3-(2-methyl
sulfonyl-benzyl)-5-(2-fluoro-
-3-methoxy-phenyl)-[1,3,5]triazinane-2,4,6-trione;
[0059]
1-(2-Amino-2-phenyl-ethyl)-3-(2,6-difluoro-benzyl)-5-(2-fluoro-phen-
yl)-[1,3,5]triazinane-2,4,6-trione;
[0060]
1-(2-Amino-2-phenyl-ethyl)-3-(2,6-dichloro-benzyl)-5-(2-fluoro-phen-
yl)-[1,3,5]triazinane-2,4,6-trione;
[0061]
1-(2-Amino-2-phenyl-ethyl)-3-(2-fluoro-6-chloro-benzyl)-5-(2-fluoro-
-phenyl)-[1,3,5]triazinane-2,4,6-trione;
[0062]
1-(2-Amino-2-phenyl-ethyl)-3-(2-fluoro-6-trifluoromethyl-benzyl)-5--
(2-fluoro-phenyl)-[1,3,5]triazinane-2,4,6-trione;
[0063]
1-(2-Amino-2-phenyl-ethyl)-3-(2-fluoro-6-methylsulfonyl-benzyl)-5-(-
2-fluoro-phenyl)-[1,3,5]triazinane-2,4,6-trione;
[0064]
1-(2-Amino-2-phenyl-ethyl)-3-(2-fluoro-benzyl)-5-(2-fluoro-phenyl)--
[1,3,5]triazinane-2,4,6-trione;
[0065]
1-(2-Amino-2-phenyl-ethyl)-3-(2-chloro-benzyl)-5-(2-fluoro-phenyl)--
[1,3,5]triazinane-2,4,6-trione;
[0066]
1-(2-Amino-2-phenyl-ethyl)-3-(2-trifluoromethyl-benzyl)-5-(2-fluoro-
-phenyl)-[1,3,5]triazinane-2,4,6-trione;
[0067]
1-(2-Amino-2-phenyl-ethyl)-3-(2-methylsulfonyl-benzyl)-5-(2-fluoro--
phenyl)-[1,3,5]triazinane-2,4,6-trione;
[0068]
1-(2-Amino-2-phenyl-ethyl)-3-(2,6-difluoro-benzyl)-5-(2-chloro-phen-
yl)-[1,3,5]triazinane-2,4,6-trione;
[0069]
1-(2-Amino-2-phenyl-ethyl)-3-(2,6-dichloro-benzyl)-5-(2-chloro-phen-
yl)-[1,3,5]triazinane-2,4,6-trione;
[0070]
1-(2-Amino-2-phenyl-ethyl)-3-(2-fluoro-6-chloro-benzyl)-5-(2-chloro-
-phenyl)-[1,3,5]triazinane-2,4,6-trione;
[0071]
1-(2-Amino-2-phenyl-ethyl)-3-(2-fluoro-6-trifluoromethyl-benzyl)-5--
(2-chloro-phenyl)-[1,3,5]triazinane-2,4,6-trione;
[0072]
1-(2-Amino-2-phenyl-ethyl)-3-(2-fluoro-6-methylsulfonyl-benzyl)-5-(-
2-chloro-phenyl)-[1,3,5]triazinane-2,4,6-trione;
[0073]
1-(2-Amino-2-phenyl-ethyl)-3-(2-fluoro-benzyl)-5-(2-chloro-phenyl)--
[1,3,5]triazinane-2,4,6-trione;
[0074]
1-(2-Amino-2-phenyl-ethyl)-3-(2-chloro-benzyl)-5-(2-chloro-phenyl)--
[1,3,5]triazinane-2,4,6-trione;
[0075]
1-(2-Amino-2-phenyl-ethyl)-3-(2-trifluoromethyl-benzyl)-5-(2-chloro-
-phenyl)-[1,3,5]triazinane-2,4,6-trione;
[0076]
1-(2-Amino-2-phenyl-ethyl)-3-(2-methylsulfonyl-benzyl)-5-(2-chloro--
phenyl)-[1,3,5]triazinane-2,4,6-trione;
[0077]
1-(2-Amino-2-phenyl-ethyl)-3-(2,6-difluoro-benzyl)-5-(3-methoxy-phe-
nyl)-[1,3,5]triazinane-2,4,6-trione;
[0078]
1-(2-Amino-2-phenyl-ethyl)-3-(2,6-dichloro-benzyl)-5-(3-methoxy-phe-
nyl)-[1,3,5]triazinane-2,4,6-trione;
[0079]
1-(2-Amino-2-phenyl-ethyl)-3-(2-fluoro-6-chloro-benzyl)-5-(3-methox-
y-phenyl)-[1,3,5]triazinane-2,4,6-trione;
[0080]
1-(2-Amino-2-phenyl-ethyl)-3-(2-fluoro-6-trifluoromethyl-benzyl)-5--
(3-methoxy-phenyl)-[1,3,5]triazinane-2,4,6-trione;
[0081]
1-(2-Amino-2-phenyl-ethyl)-3-(2-fluoro-6-methylsulfonyl-benzyl)-5-(-
3-methoxy-phenyl)-[1,3,5]triazinane-2,4,6-trione;
[0082]
1-(2-Amino-2-phenyl-ethyl)-3-(2-fluoro-benzyl)-5-(3-methoxy-phenyl)-
-[1,3,5]triazinane-2,4,6-trione;
[0083]
1-(2-Amino-2-phenyl-ethyl)-3-(2-chloro-benzyl)-5-(3-methoxy-phenyl)-
-[1,3,5]triazinane-2,4,6-trione;
[0084]
1-(2-Amino-2-phenyl-ethyl)-3-(2-trifluoromethyl-benzyl)-5-(3-methox-
y-phenyl)-[1,3,5]triazinane-2,4,6-trione;
[0085]
1-(2-Amino-2-phenyl-ethyl)-3-(2-methylsulfonyl-benzyl)-5-(3-methoxy-
-phenyl)-[1,3,5]triazinane-2,4,6-trione;
[0086]
1-(2-Amino-2-phenyl-ethyl)-3-(2,6-difluoro-benzyl)-5-(2-chloro-3-me-
thoxy-phenyl)-[1,3,5]triazinane-2,4,6-trione;
[0087]
1-(2-Amino-2-phenyl-ethyl)-3-(2,6-dichloro-benzyl)-5-(2-chloro-3-me-
thoxy-phenyl)-[1,3,5]triazinane-2,4,6-trione;
[0088]
1-(2-Amino-2-phenyl-ethyl)-3-(2-fluoro-6-chloro-benzyl)-5-(2-chloro-
-3-methoxy-phenyl)-[1,3,5]triazinane-2,4,6-trione;
[0089]
1-(2-Amino-2-phenyl-ethyl)-3-(2-fluoro-6-trifluoromethyl-benzyl)-5--
(2-chloro-3-methoxy-phenyl)-[1,3,5]triazinane-2,4,6-trione;
[0090]
1-(2-Amino-2-phenyl-ethyl)-3-(2-fluoro-6-methylsulfonyl-benzyl)-5-(-
2-chloro-3-methoxy-phenyl)-[1,3,5]triazinane-2,4,6-trione;
[0091]
1-(2-Amino-2-phenyl-ethyl)-3-(2-fluoro-benzyl)-5-(2-chloro-3-methox-
y-phenyl)-[1,3,5]triazinane-2,4,6-trione;
[0092]
1-(2-Amino-2-phenyl-ethyl)-3-(2-chloro-benzyl)-5-(2-chloro-3-methox-
y-phenyl)-[1,3,5]triazinane-2,4,6-trione;
[0093]
1-(2-Amino-2-phenyl-ethyl)-3-(2-trifluoromethyl-benzyl)-5-(2-chloro-
-3-methoxy-phenyl)-[1,3,5]triazinane-2,4,6-trione; and
[0094] 1-(2-Amino-2-phenyl-ethyl)-3-(2-methyl
sulfonyl-benzyl)-5-(2-chloro-
-3-methoxy-phenyl)-[1,3,5]triazinane-2,4,6-trione.
[0095] 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).
[0096] 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.
[0097] 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.
[0098] 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.
[0099] 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).
[0100] For example, effectiveness of a compound as a GnRH receptor
antagonist may be determined by one or more of the following
assays.
[0101] Rat Anterior Pituitary Cell Culture Assay of GnRH
Antagonists
[0102] 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.
[0103] RIA of LH and FSH
[0104] 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
[.sup.125I]-labeled rLH (.about.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.
[0105] Radio-Iodination of GnRH Peptide
[0106] The GnRH analog is labeled by the chloramine-T method. To 10
.mu.g of peptide in 20 .mu.l 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, Mass.). 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,
Calif.) 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.
[0107] GNRE Receptor Membrane Binding Assay
[0108] 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
(.about.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).
[0109] 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: 1 K i = IC 50 1 + L / K D
[0110] 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).
[0111] 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).
[0112] 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.
[0113] 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.
[0114] 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.
[0115] 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
that 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.
[0116] 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.
[0117] 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-[N-(2-PYRIDYLETHYL)-N-METHYLAMINOETHYL]-3-(3-METHOXYPHENYL)-
-5-(2,6-DIFLUOROBENZYL)ISOCYANURATE
[0118] 8
[0119] Step 1A 1-Allyl-3-(3-methoxyphenyl)isocyanurate
[0120] A solution of 1-allyl-3-(m-anisyl)urea (2.06 g, 10 mmol) and
ethyl isocyanatoformate (1.43 g, 12.5 mmol) in anhydrous
bromobenzene (15 mL) was refluxed for 1 hour. The reaction mixture
was cooled to ambient temperature and volatiles were evaporated.
The residue was partitioned between saturated NaHCO.sub.3/H.sub.2O
and dichloromethane. The organic layer was dried (sodium sulfate)
and evaporated to give desired compound (2.36 g, 85.8%), MS (CI)
m/z 276.30 (MH.sup.+).
[0121] Step 1B
1-Allyl-3-(3-methoxyphenyl)-5-(2,6-difluorobenzyl)isocyanur-
ate
[0122] A solution of compound
1-allyl-3-(3-methoxyphenyl)isocyanurate (275 mg, 1 mmol) in
anhydrous tetrahydrofurnan (5 mL) was treated with
tetrabutylammonium fluoride (1M in tetrahydrofuran, 2 mL) for 1
hour. Then 2,6-difluorobenzyl bromide (310 mg, 1.5 mmol) was added
and stirred at room temperature overnight. Volatiles were
evaporated and the residue was partitioned between saturated
NaHCO.sub.3/H.sub.2O and EtOAc. The organic layer was dried (sodium
sulfate), evaporated, and purified by flash chromatography (silica,
1:3 EtOAc/hexanes as elutant) to give compound (350 mg, 87.3%), MS
(CI) m/z 402.0 (MH.sup.+).
[0123] Step 1 C
1-Carbonylmethyl-3-(3-methoxyphenyl)-5-(2,6-difluorobenzyl-
)isocyanurate
[0124] A solution of compound
1-allyl-3-(3-methoxyphenyl)-5-(2,6-difluorob- enzyl)isocyanurate
(282 mg, 0.7 mmol) in tetrahydrofuran/water (10 mL/5 mL) was
treated with sodium periodate (750 mg, 3.5 mmol), followed by
addition of OsO.sub.4 (18 mg, 0.07 mmol). The reaction mixture was
stirred vigorously at ambient temperature for 3 hr. The reaction
mixture was partitioned between saturated NaHCO.sub.3/H.sub.2O and
EtOAc. The organic layer was dried (sodium sulfate), and evaporated
to give the title compound (201 mg, 71.3%), MS (CI) m/z 404.10
(MH.sup.+).
[0125] Step 1 D
1-[N-(2-Pyridylethyl)-N-methylamino]ethyl-3-(3-methoxyphen-
yl)-5-(2,6-difluorobenzyl)isocyanurate
[0126] A solution of
1-carbonylmethyl-3-(3-methoxyphenyl)-5-(2,6-difluorob-
enzyl)isocyanurate (81 mg, 0.2 mmol) in anhydrous dichloroethane (3
mL) was treated with 2-(2-methylaminoethyl)pyridine (55 mg, 0.4
mmol) and sodium triacetoxyborohydride (85 mg, 0.4 mmol). The
reaction mixture was stirred at ambient temperature for 3 hours.
The reaction mixture was partitioned between saturated
NaHCO.sub.3/H.sub.2O and dichloromethane. The organic layer was
washed with brine, dried (sodium sulfate), evaporated, purified by
reverse phase HPLC (C-18 column, 15-75% ACN/water) to give the
title compound, MS (CI) m/z 524.20 (MH.sup.+).
EXAMPLE 2
SYNTHESIS OF
1-[(2R)-AMINO-2-PHENYLETHYL]-3-(3-METHOXYPHENYL)-5-(2,6-DIFLU-
OROBENZYL)ISOCYANURATE
[0127] 9
[0128] Step 2A 1-(2,6-Difluorobenzyl)-3-(3-methoxyphenyl)urea
[0129] A solution of 3-methoxyphenyl isocyanate (1.0 g, 6.7 mmol)
in anhydrous dichloromethane (10 mL) was treated with
2,6-difluorobenzylamine (960 mg, 6.7 mmol) and stirred at ambient
temperature for 1 hour. The reaction mixture was filtered, and
washed with dichloromethane to give a white solid (1.47 g, 50.3%),
MS (CI) m/z 293.0 (MH.sup.+).
[0130] Step 2B
5-(2,6-Difluorobenzyl)-3-(3-methoxyphenyl)isocyanurate
[0131] A solution of 1-(2,6-difluorobenzyl)-3-(3-methoxyphenyl)urea
(292 mg, 1.0 mmol) and ethyl isocyanatoformate (143 mg, 1.25 mmol)
in anhydrous toluene (3 mL) was refluxed for 1 hour. The reaction
mixture was cooled to ambient temperature and volatiles were
evaporated. The residue was partitioned between saturated
NaHCO.sub.3/H.sub.2O and dichloromethane. The organic layer was
dried (sodium sulfate), evaporated to give the desired compound
(296 mg, 82.2%), MS (CI) m/z 362.10 (MH.sup.+).
[0132] Step 2C
1-[(2R)-Tertbutoxycarbonylamino-2-phenylethyl]-3-(3-methoxy-
phenyl)-5-(2,6-difluorobenzyl)isocyanurate
[0133] A solution of
N-(t-butyloxycarbonyl)-D-.alpha.-phenylglycinol (119 mg, 0.5 mmol)
in anhydrous tetrahydrofuran (15 mL) was treated with
5-(2,6-difluorobenzyl)-3-(3-methoxyphenyl)isocyanurate (180 mg, 0.5
mmol) and triphenylphosphine (197 mg, 0.75 mmol) at ambient
temperature, then di-tert-butylazodicarboxylate (173 mg, 0.75 mmol)
was introduced. The reaction mixture was stirred at ambient
temperature for 16 hours and volatiles were evaporated. The residue
was partitioned between saturated NaHCO.sub.3/H.sub.2O and EtOAc.
The organic layer was washed with brine, dried (sodium sulfate),
evaporated, purified by flash chromatography (silica, 3:7
EtOAc/hexanes as elutant) to give the protected compound (267 mg,
92.1%), MS (CI) m/z 481.10 (MH.sup.+-Boc).
[0134] Step 2D
1-[(2R)-Amino-2-phenylethyl]-3-(3-methoxyphenyl)-5-(2,6-dif-
luorobenzyl)isocyanurate
[0135] To a solution of
1-[(2R)-tertbutoxycarbonylamino-2-phenylethyl]-3-(-
3-methoxyphenyl)-5-(2,6-difluorobenzyl)isocyanurate (250 mg, 0.43
mmol) in dichloromethane (2 mL) was added TFA (2 mL) and the
reaction mixture was stirred at ambient temperature for 1 hour.
Volatiles were evaporated and the residue was partitioned between
saturated NaHCO.sub.3/water and EtOAc. The organic layer was dried
(sodium sulfate), evaporated, purified by reverse phase HPLC (C-18
column, 15-75% ACN/water) to give the title compound, MS (CI) m/z
481.10 (MH.sup.+). 1-[(2R)-Amino-2-phenylethyl]-3-(-
2-fluorophenyl)-5-(2,6-difluorobenzyl)isocyanurate was also
synthesized. MS (CI) m/z 469.44 (MH.sup.+).
EXAMPLE 3
1-[(2R)-AMINO-2-PHENYLETHYL]-3-(2-FLUORO-3-METHOXYPHENYL)-5-(2,6-DIFLUOROB-
ENZYL)ISOCYANURATE
[0136] 10
[0137] Step 3A
N-(2-Fluoro-3-methoxyphenyl)-N'-(2,6-difluorobenzyl)urea
[0138] A solution of diphenylphosphoryl azide (3.19 g, 2.50 mL,
11.6 mmol) in toluene (3 mL) was added dropwise to a solution of
2-fluoro-3-methoxybenzoic acid (1.70 g, 10 mmol) and triethylamine
(1.17 g, 1.62 mL, 11.6 mmol) in anhydrous bromobenzene (20 mL). The
reaction mixture was stirred at room temperature for 30 minutes and
heated at 95.degree. C. for 20 minutes. The reaction was cooled to
ambient temperature and a solution of 2,6-difluorobenzylamine (1.43
g, 10 mmol) in dichloromethane (5 mL) was added. The mixture was
stirred at ambient temperature for half of an hour, filtered, and
washed with dichloromethane and ether to give a white solid (2.36
g, 76.1%). .sup.1H NMR (DMSO-d.sub.6) .delta. 3.77 (s, 3H), 4.35
(d, J=5.4 Hz, 2H), 6.69-7.70 (m, 6H), 7.07 (br s, 1H), 8.29 (br s,
1H); MS (CI) m/z 311.0 (MH.sup.+).
[0139] Step 3B
3-(2-Fluoro-3-methoxyphenyl)-5-(2,6-difluorobenzyl)isocyanu-
rate
[0140] N-(Chlorocarbonyl)isocyanate (696 mg, 6.6 mmol) was added
slowly to a solution of
N-(2-fluoro-3-methoxyphenyl)-N'-(2,6-difluorobenzyl)urea (1.86 g,
6.0 mmol) in dichloromethane (15 mL). The reaction mixture was
stirred at ambient temperature overnight. The reaction was then
quenched with saturated NaHCO.sub.3/H.sub.2O and was extracted with
dichloromethane. The organic layer was washed with brine, dried
(sodium sulfate), evaporated to give 2.12 g, (93.0%) of white
solid. .sup.1H NMR (CDCl.sub.3) .delta. 3.90 (s, 3H), 5.22(s, 2H),
6.86-7.26 (m, 6H), 8.32 (br s, 1H); MS (CI) m/z 380.2
(MH.sup.+).
[0141] Step 3 C
1-[(2R)-Tertbutoxycarbonylamino-2-phenylethyl-3-(2-fluoro--
3-methoxyphenyl)-5-(2,6-difluorobenzyl)isocyanurate
[0142] A solution of
N-(t-butyloxycarbonyl)-D-.alpha.-phenylglycinol (48 mg, 0.2 mmol)
in anhydrous tetrahydrofuran (3 mL) was treated with
3-(2-fluoro-3-methoxyphenyl)-5-(2,6-difluorobenzyl)isocyanurate (76
mg, 0.2 mmol) and polymer-supported triphenylphosphine (250 mg,
loading 1 mmol PPh.sub.3/g) at ambient temperature.
Di-tert-butylazodicarboxylate (58 mg, 0.25 mmol) was introduced and
the reaction mixture was stirred at ambient temperature for 16
hours. The reaction mixture was filtered and the solid material was
washed with additional tetrahydrofuran. The organic filtrates were
combined and volatiles were evaporated. MS (CI) m/z 499.10
(MH.sup.+-Boc).
[0143] Step 3D
1-[(2R)-Amino-2-phenylethyl]-3-(2-fluoro-3-methoxyphenyl)-5-
-(2,6-difluorobenzyl)isocyanurate
[0144] Trifluoroacetic acid (1 mL) was added to a solution of
1-[(2R)-tertbutoxycarbonylamino-2-phenylethyl]-3-(2-fluoro-3-methoxypheny-
l)-5-(2,6-difluorobenzyl)isocyanurate in dichloromethane (1 mL) and
the reaction mixture was stirred at ambient temperature for 1 hour.
Volatiles were evaporated and the residue was partitioned between
saturated NaHCO.sub.3/water and EtOAc. The organic layer was dried
(sodium sulfate), evaporated, purified by reverse phase HPLC (C-18
column, 15-75% ACN/water). .sup.1H NMR (CDCl.sub.3) .delta. 3.82
(s, 3H), 3.99 (dd, J=13.5, 6.8 Hz, 1H), 4.35-4.49 (m, 2H), 4.98
(dd, J=25.5, 15.1 Hz, 1H), 5.20(dd, J=25.5, 14.7 Hz, 1H), 6.79-7.37
(m, 11H); MS (CI) m/z 499.10 (MH.sup.+).
[0145] The compounds in table 1 were prepared according to the
procedures outlined in example 3.
1TABLE 1 11 No. --(CR.sub.3aR.sub.3b).sub.n--NR.sub.1R.sub.2
--R.sub.5 MW Ion 3-2 12 13 508.5 509.1 3-3 14 15 532.5 533.1 3-4 16
17 480.4 481.1 3-5 18 19 468.4 469.1 3-6 20 21 498.4 499 3-7 22 23
464.4 465.4 3-8 24 25 450.4 434.2 3-9 26 27 490.5 491.5 3-10 28 29
506.5 507.5 3-11 30 31 586.6 587.2 3-12 32 33 524.5 525.4 3-13 34
35 546.6 547.4 3-14 36 37 540.5 541.3 3-15 38 39 464.4 465.5 3-16
40 41 506.5 507.4 3-17 42 43 715.5 715.4 3-18 44 45 368.3 369.4
3-19 46 47 394.4 395.3 3-20 48 49 504.5 505.4 3-21 50 51 428.5
429.3 3-22 52 53 450.5 451.4 3-23 54 55 619.4 618.6 3-24 56 57
430.4 431.1 3-25 58 59 494.4 495.1 3-26 60 61 508.5 509.1 3-27 62
63 424.3 425.1 3-28 64 65 410.3 411 3-29 66 67 506.4 507.3 3-30 68
69 500.4 501.1 3-31 70 71 424.3 425 3-32 72 73 466.4 467.1 3-33 74
75 560.5 561.2 3-33 76 77 546.5 547.1 3-34 78 79 484.4 485.1 3-35
80 81 530.4 531.1 3-36 82 83 544.5 545.1 3-37 84 85 576.5 577.2
3-38 86 87 456.3 457.1 3-39 88 89 442.3 443 3-40 90 91 538.5 539.2
3-41 92 93 532.4 533.1 3-42 94 95 456.3 457.1 3-43 96 97 498.4
499.1 3-44 98 99 482.4 483.3 3-45 100 101 498.4 499.1 3-46 102 103
578.5 579.2 3-47 104 105 571.5 572 3-48 106 107 608.5 609.2 3-49
108 109 404.3 405.1 3-50 110 111 494.4 495.3 3-51 112 113 418.3
419.1 3-52 114 115 460.4 461.2 3-53 116 117 444.4 445.1 3-54 118
119 460.4 461.1 3-55 120 121 554.6 555.1 3-56 122 123 540.5 541.1
3-57 124 125 478.5 479.1 3-58 126 127 524.5 425.2 3-59 128 129
538.5 539.2 3-60 130 131 480.4 481 3-61 132 133 570.5 571.2 3-62
134 135 512.5 513 3-63 136 137 535 535.1 3-64 138 139 516.4 517.1
3-64 140 141 500.5 501 3-65 142 143 522.5 523.2 3-66 144 145 380.4
381.1 3-67 146 147 464.4 465.1 3-68 148 149 464.4 465.1
[0146] NMR Data
[0147] 3-2 .sup.1H NMR (CDCl.sub.3) .delta. 2.65 (br s, 2H), 4.06
(dd, J=14.5, 2.6 Hz, 1H), 4.43 (dd, J=14.5, 10.6 Hz, 1H), 4.63 (dd,
J=10.6, 2.6 Hz, 1H), 5.15 (s, 2H), 6.83-8.05 (m, 11H); MS (CI) m/z
509.1 (MH.sup.+).
EXAMPLE 4
1-[(2R)-AMINO-2-PHENYLETHYL]-3-(2-FLUORO-3-METHOXYPHENYL)-5-(2,6-DICHLOROB-
ENZYL)ISOCYANURATE
[0148] 150
[0149] Step 4A
1-Azido-N-(2R)-tertbutoxycarbonylamino-2-phenylethane
[0150] A solution of
N-(t-butyloxycarbonyl)-D-.alpha.-phenylglycinol-O-mes- ylate (3.15
g, 10 mmol) in anhydrous DMF (15 mL) was treated with sodium azide
(3.25 g, 50 mmol) and the resulting mixture was heated at
80.degree. C. for 4 hours. After cooling, the mixture was
partitioned between EtOAc and water. The organic layer was washed
with brine, dried and evaporated to give azido compound as a white
solid (2.31 g, 88.2%). .sup.1H NMR (CDCl.sub.3) .delta. 1.43 (s,
9H), 3.63 (d, J=4.3 Hz, 2H), 4.87 (br s, 1H), 5.07 (d, J=4.3 Hz,
1H), 7.26-7.39 (m, 5H).
[0151] Step 4B
1-Amino-N-(2R)-tertbutoxycarbonylamino-2-phenylethane
[0152] A suspension of lithium aluminum hydride (270 mg, 7.1 mmol)
in tetrahydrofuran (16 mL) was cooled down to -40.degree. C. and a
solution of 1-azido-N-(2R)-tertbutoxycarbonylamino-2-phenylethane
(1.03 g, 3.9 mmol) in tetrahydrofuran (10 mL) was added dropwise.
The reaction mixture was gradually warmed from -40.degree. C. to
-10.degree. C. in 3 hours. The mixture was then cooled to
-50.degree. C., quenched with water and extracted with EtOAc. The
organic layer was washed with brine, filtered through celite, dried
(sodium sulfate), and evaporated to give the desired diamine (886
mg, 96.2%). MS (CI) m/z 311.0 (MH.sup.+).
[0153] Step 4C
N-[(2R)-Tertbutoxycarbonylamino-2-phenylethyl]-N'-(2-fluoro-
-3-methoxyphenyl)urea
[0154] A solution of diphenylphosphoryl azide (1.266 g, 1.0 mL, 4.6
mmol) in toluene (3 mL) was added dropwise to a solution of
2-fluoro-3-methoxybenzoic acid (680 mg, 4 mmol) and triethylamine
(468 mg, 0.65 mL, 4.6 mmol) in anhydrous toluene (20 mL). The
reaction mixture was stirred at room temperature for 30 minutes,
and then heated at 95.degree. C. for 20 minutes. The reaction was
cooled down to ambient temperature and a solution of
1-amino-N-(2R)-tertbutoxycarbonylamino-2-ph- enylethane (886 mg,
3.75 mmol) in dichloromethane (5 mL) was added. The reaction
mixture was stirred at ambient temperature for half of an hour,
filtered, and washed with dichloromethane and ether to give white
solid (1.04 g, 68.8%). MS (CI) m/z 304.10 (MH.sup.+-Boc).
[0155] Step 4D
1-[(2R)-Tertbutoxycarbonylamino-2-phenylethyl]-3-(2-fluoro--
3-methoxyphenyl)isocyanurate
[0156] A solution of
N-[(2R)-tertbutoxycarbonylamino-2-phenylethyl]-N'-(2--
fluoro-3-methoxyphenyl)urea (1.0 g, 2.48 mmol) and ethyl
isocyanatoformate (356 mg, 3.1 mmol) in anhydrous bromobenzene (4
mL) was refluxed for 1 hour. The reaction mixture was cooled down
to ambient temperature and volatiles were evaporated. The residue
was partitioned between saturated NaHCO.sub.3/H.sub.2O and
dichloromethane. The organic layer was dried (sodium sulfate),
evaporated, and purified on a silica gel column, eluted with 1:1
EtOAc/hexanes to give the desired compound (937 mg, 80.1%). .sup.1H
NMR (CDCl.sub.3) .delta. 1.37 (s, 9H), 3.92 (s, 3H), 3.86-4.03 (m,
2H), 4.22-4.34 (m, 1H), 5.22 (br s, 1H), 6.86-7.35 (m, 8H); MS (CI)
m/z 373.10 (MH.sup.+-Boc).
[0157] Step 4E
1-[(2R)-Amino-2-phenylethyl]-3-(2-fluoro-3-methoxyphenyl)-5-
-(2,6-dichlorobenzyl)isocyanurate
[0158] A solution of
1-[(2R)-tertbutoxycarbonylamino-2-phenylethyl]-3-(2-f-
luoro-3-methoxyphenyl)isocyanurate (47.2 mg, 0.1 mmol) in anhydrous
DMF (2 mL) was treated with potassium carbonate (35 mg, 0.25 mmol)
and 2,6-dichlorobenzyl bromide (28.8 mg, 0.12 mmol) at ambient
temperature. The reaction mixture was stirred at ambient
temperature for 16 hours and was partitioned between
dichloromethane and water. The organic layer was dried and
evaporated to give BOC-protected compound, which was treated with
1:1 TFA/DCM (2 mL) for 1 hour. The reaction was evaporated,
purified by reverse phase HPLC (C-18 column, 15-75% ACN/water).
.sup.1H NMR (CDCl.sub.3) .delta. 3.82 (s, 3H), 4.03 (t, J=15.5 Hz,
1H), 4.36-4.48 (m, 2H), 5.16 (dd, J=25.5, 15.1 Hz, 1H), 5.38 (dd,
J=25.5, 14.6 Hz, 1H), 6.84-7.35 (m, 11H), MS (CI) m/z 531.0
(MH.sup.+).
[0159] The compounds in table 2 were prepared according to the
procedures outlined in example 4.
2TABLE 2 151 No. --(CR.sub.3aR.sub.3b).sub.n--NR.sub.1R.sub.2
--R.sub.4 --R.sub.5 MW Ion 4-2 H 2,6-di-F-Bz butyl 311.3 312.3 4-3
H butyl allyl 225.3 226.4 4-4 H cyclohexyl allyl 251.3 252.2 4-5
152 2,6-di-F-Bz 2,6-di-F-Ph 539.5 540.1 4-6 H.sub.2NCH.sub.2CH2
allyl butyl 268.3 269.2 4-7 153 Bz allyl 498.6 499.2 4-8 154 Bz
butyl 430.5 431.1 4-9 155 2,6-di-F-Bz Ph 450.4 451.1 4-10 156
2,6-di-F-Bz 4-CF.sub.3-Ph 518.4 519.1 4-11 157 158 159 628.7 623.9
4-12 160 4-OMe-Bz 161 632.7 633.3 4-13 162 4-OMe-Bz 4-OCF.sub.3-Bz
612.6 613.2 4-14 163 4-OMe-Bz 4-t-butyl-Bz 540.7 541.2 4-15 164
4-OCF.sub.3-Bz 165 596.6 597.2 4-16 166 4-OMe-Bz 167 620.7 621.3
4-17 168 2,4,6-tri-Me-Bz 2-F-3-OMe-Ph 504.6 4-18 169
2-F-6-CF.sub.3-Bz 2-F-3-OMe-Ph 548.5 549.1 4.19 170
Cyclopropyl-CH.sub.2-- 2-F-3-OMe-Ph 426.4 427.1 4-20 171 2-Me-Bz
2-F-3-OMe-Ph 476.5 477.1 4-21 172 2-OMe-Bz 2-F-3-OMe-Ph 492.5 493.1
4-22 173 Cyclohexyl-CH.sub.2-- 2-F-3-OMe-Ph 468.5 469.1 4-23 174
2-CF.sub.3-Bz 2-F-3-OMe-Ph 530.5 531.1 4-24 175 2-Cl-6-F-Bz
2-F-3-OMe-Ph 514.9 515.1 4-25 176 3,5-Di-OMe-Bz 2-F-3-OMe-Ph 522.5
523.1 4-26 177 178 2-F-3-OMe-Ph 520.5 521.3 4-27 179 180
2-F-3-OMe-Ph 526.5 527.3 4-28 181 182 2-F-3-OMe-Ph 546.6 547.2 4-29
183 184 2-F-3-OMe-Ph 548.5 549.2 4-30 185 186 2-F-3-OMe-Ph 548.5
549.2 4-31 187 188 2-F-3-OMe-Ph 548.5 549.2 4-32 189 190
2-F-3-OMe-Ph 548.5 549.2 4-33 191 192 2-F-3-OMe-Ph 520.5 521.2 4-34
193 194 2-F-3-OMe-Ph 532.4 532.1 4-35 195 196 2-F-3-OMe-Ph 487.5
488.1
[0160] NMR Data
[0161] 4-2 .sup.1H NMR (CDCl.sub.3) .delta. 0.930 (t, J=7.35 Hz,
3H), 1.30-1.40 (m, 2H), 1.57-1.66 (m, 2H), 3.82-3.87 (d, J=7.8 Hz,
2H), 5.18 (s, 2H), 6.86-6.92 (m, 2H), 7.20-7.30(m, 1H); MS (CI) m/z
312.30 (MH.sup.+).
[0162] 4-3 .sup.1H NMR (CDCl.sub.3) .delta. 0.95 (t, J=7.35 Hz,
3H), 1.30-1.43 (m, 2H), 1.58-1.69 (m, 2H), 3.87 (t, J=7.65 Hz, 2H),
4.47 (d, J=4.5 Hz, 2H), 5.24-5.35 (m, 2H), 5.81-5.94 (m, 1H), 8.76
(br s, 1H); MS (CI) m/z 226.4 (MH.sup.+).
[0163] 4-4 .sup.1H NMR (CDCl.sub.3) .delta. 1.17-1.41 (m, 2H),
1.66-1.69 (m, 4H), 1.82-1.87 (m, 2H), 2.20-2.36 (m, 2H), 4.45 (d,
J=6.0 Hz, 2H), 4.52-4.61 (m, 1H), 5.25 (dd, J=9.9, 1.2 Hz, 1H),
5.31 (dd, J=17.1, 1.2 Hz, 1H), 5.80-6.00 (m, 1H), 8.40 (br s, 1H);
MS (CI) m/z 252.20 (MH.sup.+).
[0164] 4-6 .sup.1H NMR (300 MHz, CDCl.sub.3) .delta. 0.95 (t, 3H),
1.20-60 (m, 4H), 3.30-3.40 (b, 2H), 3.95 (t, 2H), 4.18-4.23(b, 2H),
4.45 (d, 2H), 5.10-5.40 (m, 2H), 5.80-5.95 (m, 1H); MS (CI) m/z
269.20 (MH.sup.+).
[0165] 4-7 .sup.1H NMR (300 MHz, CDCl.sub.3) .delta. 3.00 (d, 2H),
3.90 (br s, 1H), 4.00 (d, 1H), 4.10-4.20 (m, 3H), 4.45 (d, 1H),
4.95-5.05 (m, 4H), 5.05-5.17 (m, 2H), 5.60-5.80 (m, 1H), 6.95 (d,
2H), 7.10-7.50 (m, 12H); MS (CI) m/z 499.20 (MH.sup.+).
[0166] 4-8 .sup.1H NMR (CDCl.sub.3) .delta. 0.95 (t, 3H), 1.10-1.20
(m, 2H), 1.40-1.60 (m, 2H), 3.89 (t, 2H), 3.97 (d, 1H), 4.25 (dd,
1H), 4.65 (d, 1H), 5.05-5.17 (m, 2H), 6.95 (t, 2H), 7.20-7.45 (m,
6H); MS (CI) m/z 431.10 (MH.sup.+).
[0167] 4-9 .sup.1H NMR (CDCl.sub.3) .delta. 3.97 (d, J=12.00 Hz,
1H), 4.35 (dd, J=14.1, 9.9 Hz, 1H), 4.52 (d, J=9.3 Hz, H), 5.06 (s,
2H), 6.82-6.88 (m, 2H), 7.19-7.37 (m, 8H), 7.70 (d, J=8.70 Hz, 2H);
MS (CI) m/z 519.10 (MH.sup.+).
[0168] 4-11 .sup.1H NMR (CDCl.sub.3) .delta. 2.25-2.40 (m, 2H),
2.40-2.50 (m, 2H), 2.60-2.70 (m, 1H), 3.30-3.40 (m, 2H), 3.50-3.60
(m, 2H), 3.90-4.20 (m, 8H), 4.42 (s, 2H), 7.10-7.40 (m, 20H); MS
(CI) m/z 629.30 (MH.sup.+).
[0169] 4-12 .sup.1H NMR (CDCl.sub.3) .delta. 1.40-1.80 (m, 4H),
2.30-2.45 (m, 2H), 3.40 (d, 1H), 3.58-3.65 (m, 1H), 3.77 (s, 3H),
3.90-4.30 (m, 5H), 4.45 (s, 2H), 4.80-4.95 (m, 2H), 6.84 (d, 2H),
7.16-7.38 (m, 17H); MS (CI) m/z 633.30 (MH.sup.+).
[0170] 4-13 .sup.1H NMR (CDCl.sub.3) .delta. 1.82-2.00 (m, 2H),
2.30-2.40 (m, 2H), 3.42 (d, 2H), 3.65-3.90 (m, 5H), 4.05-4.50 (m,
3H), 4.93-4.98 (m, 4H), 6.81 (t, 2H), 7.12-7.44 (m, 11H); MS (CI)
m/z 613.20 (MH.sup.+).
[0171] 4-14 .sup.1H NMR (CDCl.sub.3) .delta. 1.28 (s, 1H), 3.10 (d,
J=7.5 Hz, 2H), 3.77 (s, 3H), 3.91-4.13 (m, 2H), 4.48-4.93 (m, 7H),
6.82 (d, J=9.0 Hz, 2H), 7.09-7.37 (m, 10H); MS (CI) m/z 541.20
(MH.sup.+).
[0172] 4-15 .sup.1H NMR (CDCl.sub.3) .delta. 1.20-1.40 (m, 5H),
2.20-2.35 (m, 1H), 2.60-2.80 (m, 1H), 3.30-3.42 (m, 1H), 3.80-3.95
(m, 2H), 4.30-4.47 (m, 3H), 4.65 (d, 2H), 4.87-4.92 (m, 2H),
7.07-7.39 (m, 14H); MS (CI) m/z 597.20 (MH.sup.+).
[0173] 4-16 .sup.1H NMR (300 MHz, CDCl.sub.3) .delta. 1.39 (d, 3H),
2.25-2.40 (m2H), 3.35 (t, 1H), 3.75 (s, 3H), 3.75-3.95 (m, 5H),
4.15-4.22 (m, 1H), 4.45 (d, 1H), 4.66 (d, 1H), 4.78 (s, 2H), 6.78
(d, 2H), 7.13-7.32 (m, 17H); MS (CI) m/z 621.30 (MH.sup.+).
[0174] 4-17 .sup.1H NMR (CDCl.sub.3) .delta. 3.81-3.84(d, J=7.2 Hz,
3H), 4.05-4.12 (m, 2H), 4.40-4.58 (m, 2H), 5.03 (s, 1H), 5.38 (d,
J=4.2 Hz, 1H), 6.784-7.63 (m, 1H).
[0175] 4-20 .sup.1H NMR (CDCl.sub.3) .delta. 2.305-2.416 (d, J=33.3
Hz, 3H), 3.802-3.830 (d, J=8.4 Hz, 3H), 3.486 (s, 0.5H),
3.972-4.033(m, 1H), 4.351-4.435(m, 2H), 4.824-4.978 (m, 1H), 5.097
(s, 1H), 6.892-7.345 (m, 11H).
[0176] 4-29 .sup.1H NMR (CDCl.sub.3) .delta. 3.814-3.838(d, J=7.2
Hz, 3H), 4.054-4.125(m, 2H), 4.439-4.584(m, 2H), 5.033 (s, 1H),
5.190(d, J=4.2 Hz, 1H), 6.775-7.632(m, 11H).
EXAMPLE 5
1-[(2R)-AMINO-2-PHENYLETHYL]-3-(2-FLUORO-3-METHOXYPHENYL)-5-(CYCLOBUTYLMET-
HYL)ISOCYANURATE
[0177] 197
[0178] Step 4A
1-[(2R)-Amino-2-phenylethyl]-3-(2-fluoro-3-methoxyphenyl)-5-
-(cyclobutylmethyl)isocyanurate
[0179] A solution of
1-[(2R)-tert-butoxycarbonylamino-2-phenylethyl]-3-(2--
fluoro-3-methoxyphenyl)isocyanurate (30 mg, 0.064 mmol) in
anhydrous tetrahydrofuran (1.0 mL) was treated with
cyclobutanemethanol (6.0 uL, 0.064 mmol), Ph.sub.3P (42.4 mg, 0.13
mmol) and di-t-butyl-azodicarboxyla- te (21.97 mg, 0.95 mmol) at
ambient temperature. The reaction mixture was stirred at ambient
temperature for 16 hours. The mixture was filtered through a 0.2
.mu.m filter and was evaporated to give protected compound, which
was treated with 1:1 TFA/DCM (2 mL) for 1 hour. The reaction
mixture was evaporated and purified by reverse phase HPLC (C-18
column, 15-75% ACN/water). .sup.1H NMR (CDCl.sub.3) .delta.
1.69-2.06 (m, 6H), 2.54-2.72 (m, 1H), 3.70-3.72 (m, 1H), 3.82-3.84
(d, J=7.8 Hz, 3H), 3.95-4.01 (m, 2H), 4.42-4.44 (m, 2H), 6.76-7.41
(m, 8H), MS (CI) m/z 441.1 (MH.sup.+).
[0180] The compounds in table 3 were prepared according to the
procedures outlined in example 5.
3TABLE 3 198 No. --R.sub.4 MW Ion 5-2 199 440.4 441.1 5-3 200 454.5
455.2 5-4 201 496.9 497.1 5-5 202 512.5 513.2 5-6 203 512.5 513.2
5-7 204 450.4 451.1 5-8 205 466.5 467.1 5-9 206 476.5 477.1 5-10
207 477.5 478.1 5-11 208 491.5 491.1 5-12 209 506.5 507.1 5-13 210
508.6 509.3 5-14 211 514.6 515.3 5-15 212 519.5 520.3 5-16 213
538.6 539.3 5-17 214 552.6 553.3 5-18 215 497.9 498.1 5-19 216
521.5 522.2 5-20 217 598.5 599.2
[0181] NMR Data
[0182] 5-11 .sup.1H NMR (CDCl.sub.3) .delta.2.00 (m, 2H),
2.603-2.689(m, 2H), 3.83 (s, 1H), 3.949-3.980 (m, 3H),
4.379-4.508(m, 2H), 6.80-7.388(m, 13H), MS (CI) m/z 491.1
(MH.sup.+).
[0183] 5-18 .sup.1H NMR (CDCl.sub.3) .delta.3.832-3.849(d, J=5.1
Hz, 3H), 4.093-4.166(m, 1H), 4.472-4.634(m, 2H), 5.129-5.218(m,
2H), 6.808-8.394(m, 11H).
EXAMPLE 6
1-(2-CYCLOPENTYLAMINO-2-METHYL-PROPYL)-3-(2-FLUORO-3-METHOXY-PHENYL)-5-(2--
FLUORO-6-TRIFLUOROMETHYL-BENZYL)-[1,3,5]TRIAZINANE-2,4,6-TRIONE
[0184] 218
[0185] Step 6A
1-(2-Amino-2-methyl-propyl)-3-(2-fluoro-3-methoxy-phenyl)-u-
rea
[0186] To a 250 mL round bottomed flask was charged 2.0 g of
2-fluoro-3-methoxy-benzoic acid in 80 mL of anhydrous toluene.
Triethylamine (1.15 eq, 1.88 mL) was added after drying over
molecular sieves. After dropwise addition of diphenylphosphoryl
azide (1.15 eq, 2.8 mL) in 12 mL toluene, the reaction was stirred
for 20 minutes at ambient temperature. The reaction was heated to
95.degree. C., then stirred for 20 minutes. After the reaction was
cooled to ambient temperature, 1,2-diamino-2-methyl-propane (1.15
eq, 1.42 mL) was added in 20 mL CH.sub.2Cl.sub.2. Urea formation
was complete in 2 hours. The reaction mixture was diluted with 100
mL H.sub.2O and extracted with EtOAc (3.times.100 mL). The combined
organics were washed with brine (1.times.100 mL), dried over
Na.sub.2SO.sub.4, filtered, and concentrated in vacuo. The residue
was filtered over a pad of silica (65.times.40 mm), impurities
eluted with 100% EtOAc, the product eluted with 50%
MeOH/CH.sub.2Cl.sub.2. The eluent was concentrated in vacuo to
yield 1.85 g (62%) of
1-(2-Amino-2-methyl-propyl)-3-(2-fluoro-3-methoxy-phenyl)-urea as a
brown solid. .sup.1H NMR (CDCl.sub.3) .delta.: 7.61 (dt, 1H), 7.0
(dt, 1H), 6.61 (dt, 1H), 5.75 (bs, 1H), 3.87 (s, 3H), 3.14 (d, 2H),
1.12 (s, 6H); MS (CI) m/z 256 (MH.sup.+).
[0187] Step 6B
{2-[3-(2-Fluoro-3-methoxy-phenyl)-ureido]-1,1-dimethyl-ethy-
l}-carbamic acid tert-butyl ester
[0188]
1-(2-Amino-2-methyl-propyl)-3-(2-fluoro-3-methoxy-phenyl)-urea (1.8
g) was dissolved in 50 mL of CH.sub.2Cl.sub.2 in a 100 mL round
bottomed flask. Di-tert-butyl dicarbonate (1.3 eq, 2.0 g) was added
and the reaction was complete after two hours by thin layer
chromatography. The reaction was diluted with 50 mL of H.sub.2O,
and the organic layer was collected. The aqueous layer was
extracted with CH.sub.2Cl.sub.2 (2.times.50 mL). The combined
organics were washed with brine, dried over Na.sub.2SO.sub.4,
filtered, and concentrated in vacuo. The residue was purified via
flash column chromatography and eluted with 30% EtOAc/hexanes to
yield 1.4 g (56%). .sup.1H NMR (CDCl.sub.3) .delta.: 7.64 (dt, 1H),
6.98 (dt, 1H), 6.91 (bs, 1H), 6.63 (dt, 1H), 6.05 (bs, 1H), 3.86
(s, 3H), 3.42 (d, 2H), 1.42 (s, 9H), 1.24 (s, 6H).
[0189] Step 6C
{2-[3-(2-Fluoro-3-methoxy-phenyl)-2,4,6-trioxo-[1,355]triaz-
inan-1-yl]-1,1-dimethyl-ethyl}-carbamic acid tert-butyl ester
[0190] To 20 mL of anhydrous CH.sub.2Cl.sub.2 was added 0.93 g of
{2-[3-(2-Fluoro-3-methoxy-phenyl)-ureido]-1,1-dimethyl-ethyl}-carbamic
acid tert-butyl ester and triethylamine (1.0 eq, 0.36 mL).
N-(chlorocarbonyl) isocyanate (1.0 eq, 0.11 mL) was added to the
solution in one portion. The reaction stalled after one hour, so
additional triethylamine (1.0 eq, 0.36 mL) and N-(chlorocarbonyl)
isocyanate was added. The reaction was determined to be complete by
LC/MS after two hours. Excess isocyanate was quenched with the
dropwise addition of H.sub.2O (1 mL). The reaction was then diluted
with 20 mL H.sub.2O, and the organic layer was collected. The
aqueous layer was extracted with CH.sub.2Cl.sub.2 (2.times.20 mL).
The combined organics were washed with brine, dried over
Na.sub.2SO.sub.4, filtered, and concentrated in vacuo. The residue
was purified via flash column chromatography and eluted with 60%
EtOAc/hexanes to yield 0.65 g (59%). R.sub.f=0.66 in 60%
EtOAc/hexanes. .sup.1H NMR (CDCl.sub.3) .delta.: 7.19 (dt, 1H),
7.09 (dt, 1H), 6.90 (dt, 1H), 5.15 (ds, 1H), 4.1 (q, 2H), 3.92 (s,
3H), 1.42 (s, 9H), 1.38 (d, 6H).
[0191] Step 6D
{2-[3-(2-Fluoro-3-methoxy-phenyl)-5-(2-fluoro-6-trifluorome-
thyl-benzyl)-2,4,6-trioxo-f
1.35]triazinan-1-yl]-1,1-dimethyl-ethyl}-carba- mic acid tert-butyl
ester
[0192]
{2-[3-(2-Fluoro-3-methoxy-phenyl)-2,4,6-trioxo-[1,3,5]triazinan-1-y-
l]-1,1-dimethyl-ethyl}-carbamic acid tert-butyl ester (370 mg) was
dissolved in 7 mL of anhydrous DMF. To this solution was added
K.sub.2CO.sub.3 (2.0 eq, 226 mg) and
2-fluoro-6-(trifluoromethyl)benzyl bromide (1.0 eq, 222 mg). The
reaction was complete by LC/MS after 14 hours. The DMF was removed
in vacuo. The residue was partitioned between EtOAc and H.sub.2O.
The EtOAc layer was collected and the aqueous layer was extracted
with EtOAc (2.times.20 mL). The combined organics were washed with
brine, dried over Na.sub.2SO.sub.4, filtered, and concentrated in
vacuo. The residue was purified via flash column chromatography and
eluted with 30% EtOAc/hexanes to yield 399 mg (76%). R.sub.f=0.36
in 30% EtOAc/hexanes. .sup.1H NMR (CDCl.sub.3) .delta.: 7.51 (d,
1H), 7.39 (m, 1H), 7.27 (d, 1H), 7.15 (dt, 1H), 7.06 (dt, 1H), 6.88
(dt, 1H), 5.42 (q, 2H), 5.15 (bs, 1H), 4.14 (q, 2H), 3.9 (s, 3H),
1.4 (s, 9H), 1.39 (d, 6H).
[0193] Step 6E
1-(2-Amino-2-methyl-propyl)-3-(2-fluoro-3-methoxy-phenyl)-5-
-(2-fluoro-6-trifluoromethyl-benzyl)isocyanurate
[0194]
{2-[3-(2-Fluoro-3-methoxy-phenyl)-2,4,6-trioxo-[1,3,5]triazinan-1-y-
l]-1,1-dimethyl-ethyl}-carbamic acid tert-butyl ester (360 mg) was
dissolved in 5 mL CH.sub.2Cl.sub.2 and 5 mL trifluoroacetic acid.
The reaction was determined to be complete by LC/MS after stirring
for one hour at ambient temperature. The solvent was removed in
vacuo, and the residue was partitioned between a saturated
NaHCO.sub.3 solution and CH.sub.2Cl.sub.2. The organic layer was
separated and washed with brine, dried over Na.sub.2SO.sub.4,
filtered, and concentrated in vacuo. The residue was filtered over
a plug of silica (33.times.30 mm) and the product eluted with 10%
MeOH/CH.sub.2Cl.sub.2. The filtrate was concentrated in vacuo to
yield 283 mg (94%) of a white foamy solid. R.sub.f=0.43 in 10%
MeOH/CH.sub.2Cl.sub.2. .sup.1H NMR (CDCl.sub.3) .delta.: 7.51 (d,
1H), 7.39 (m, 1H), 7.27 (d, 1H), 7.15 (dt, 1H), 7.05 (dt, 1H), 6.91
(dt, 1H), 5.4 (s, 2H), 4.0 (s, 2H), 3.87 (s, 3H), 3.5 (bs, 2H),
1.22 (d, 6H); MS (CI) m/z 501 (MH.sup.+).
[0195] Step 6F
1-(2-Cyclopentylamino-2-methyl-propyl)-3-(2-fluoro-3-methox-
y-phenyl)-5-(2-fluoro-6-trifluoromethyl-benzyl)isocyanurate
[0196]
1-(2-Amino-2-methyl-propyl)-3-(2-fluoro-3-methoxy-phenyl)-5-(2-fluo-
ro-6-trifluoromethyl-benzyl)isocyanurate (30 mg) was dissolved in 1
mL of 1,2-dichloroethane. Cyclopentanone (2.0 eq, 11 .mu.L) was
added and the solution was stirred for 10 minutes, at which time
Na(OAc).sub.3BH (2.0 eq, 25 mg) was added. The resulting solution
stirred at ambient temperature overnight and additional
cyclopentanone (10 equiv, 55 .mu.L) and Na(OAc).sub.3BH (2.0 eq, 25
mg) were added. The reaction was heated to 50.degree. C. for three
days. The reaction was concentrated under N.sub.2 gas, diluted to
with 1 mL of methanol, and filtered through a 0.45 .mu.m filter.
The product was purified by RP-HPLC to yield 5.4 mg (13%). MS (CI)
m/z 569 (MH.sup.+).
[0197] The compounds in table 4 were prepared according to the
procedures outlined in example 6.
4TABLE 4 219 No. --(CR.sub.3aR.sub.3b).sub.n--NR.sub.1R.sub.2 MW
Ion 6-2 220 568.5 569 6-3 221 591.5 591 6-4 222 657.6 658 6-5 223
583.5 584 6-6 224 633.6 634 6-7 225 556.5 557 6-8 226 605.6 606
[0198] 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.
[0199] 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.
* * * * *