U.S. patent application number 12/833758 was filed with the patent office on 2010-11-04 for acylated amino acid amidyl pyrazoles and related compounds.
Invention is credited to Darren Dressen, Albert Garofalo, Ashley Guinn, Scott A. Jenkins, Lee H. Latimer, Michael A. Pleiss, Jennifer Sealy, John Tucker, Jay S. Tung, David W.G. Wone, Jing Wu.
Application Number | 20100280066 12/833758 |
Document ID | / |
Family ID | 34102630 |
Filed Date | 2010-11-04 |
United States Patent
Application |
20100280066 |
Kind Code |
A1 |
Tung; Jay S. ; et
al. |
November 4, 2010 |
ACYLATED AMINO ACID AMIDYL PYRAZOLES AND RELATED COMPOUNDS
Abstract
This invention is directed to acylated amino acid amidyl
pyrazoles and related compounds of Formula I. The invention is also
directed to a pharmaceutical formation comprising such compound or
in a pharmaceutically acceptable salt form thereof. The invention
is further directed to a method for inhibiting .beta.-amyloid
peptide release and/or synthesis, a method for inhibiting
.gamma.-secretase activity, and a method for treating neurological
disorders associated with .beta.-amyloid peptide production. The
method comprises administering to a host a pharmaceutical
formulation comprising an effective amount of a compound of Formula
I. The compounds of Formula I are useful in the prevention and
treatment of Alzheimer's disease.
Inventors: |
Tung; Jay S.; (Belmont,
CA) ; Latimer; Lee H.; (Oakland, CA) ; Wu;
Jing; (Redwood City, CA) ; Garofalo; Albert;
(South San Francisco, CA) ; Pleiss; Michael A.;
(Sunnyvale, CA) ; Dressen; Darren; (Fremont,
CA) ; Guinn; Ashley; (Santa Monica, CA) ;
Jenkins; Scott A.; (Tucson, AZ) ; Sealy;
Jennifer; (Oakland, CA) ; Tucker; John; (San
Diego, CA) ; Wone; David W.G.; (Newark, CA) |
Correspondence
Address: |
HOWREY LLP-CA
C/O IP DOCKETING DEPARTMENT, 2941 FAIRVIEW PARK DRIVE, SUITE 200
FALLS CHURCH
VA
22042-2924
US
|
Family ID: |
34102630 |
Appl. No.: |
12/833758 |
Filed: |
July 9, 2010 |
Related U.S. Patent Documents
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Application
Number |
Filing Date |
Patent Number |
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10559823 |
Mar 1, 2007 |
7759382 |
|
|
PCT/US2004/018202 |
Jun 4, 2004 |
|
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12833758 |
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60476369 |
Jun 5, 2003 |
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Current U.S.
Class: |
514/307 ;
435/375; 514/326; 514/341; 514/407; 546/146; 546/211; 546/275.4;
548/364.1; 548/364.4; 548/365.7; 548/371.7 |
Current CPC
Class: |
C07D 401/12 20130101;
C07D 405/04 20130101; C07D 401/06 20130101; C07D 401/04 20130101;
C07D 231/40 20130101; A61P 43/00 20180101; C07D 405/12 20130101;
C07D 409/12 20130101; A61P 25/28 20180101 |
Class at
Publication: |
514/307 ;
548/371.7; 546/275.4; 546/146; 548/365.7; 548/364.4; 546/211;
548/364.1; 514/407; 514/341; 514/326; 435/375 |
International
Class: |
A61K 31/4725 20060101
A61K031/4725; C07D 231/38 20060101 C07D231/38; C07D 401/12 20060101
C07D401/12; C07D 407/04 20060101 C07D407/04; C07D 401/04 20060101
C07D401/04; C07D 403/12 20060101 C07D403/12; A61P 25/28 20060101
A61P025/28; A61K 31/415 20060101 A61K031/415; A61K 31/4439 20060101
A61K031/4439; A61K 31/454 20060101 A61K031/454; C12N 5/07 20100101
C12N005/07 |
Claims
1. A compound of Formula I: ##STR00154## or a pharmaceutically
acceptable salt wherein R is substituted or unsubstituted aryl,
heteroaryl, cycloalkyl, heterocyclic, alkoxy, ##STR00155##
cycloalkoxy, aryloxy, heteroaryloxy, alkylamino, cycloaklylamino,
arylamino, heteroarylamino; or R is wherein X' and X'' are each
independently hydrogen, hydroxy or fluoro, provided when one of X'
and X'' is fluoro, the other is not hydroxy; or X' and X'' together
form an oxo group, Z is selected from the group consisting of
alkyl, nitrogen, oxygen, sulfur and a bond covalently linking
R.sub.1 to --CX'X''-- R.sub.1 is selected from the group consisting
of hydrogen, substituted or unsubstituted alkyl, alkenyl, aryl,
cycloalkyl, cycloalkenyl, heteroaryl, and heterocyclic; R.sub.2 is
selected from the group consisting of hydrogen, C.sub.1-C.sub.4
alkyl, --COOR.sub.2a, and --COR.sub.2a wherein R.sub.2a is
hydrogen, C.sub.1-4 alkyl, cycloalkyl, or heterocycle; R.sub.3 is
H, substituted or unsubstituted linear alkyl, branched alkyl,
cycloalkyl, or phenyl; R.sub.5 is --Y--R.sub.6, wherein Y is
substituted or unsubstituted alkyl, alkenyl, aryl, cycloalkyl,
cycloalkenyl, heteroaryl, or heterocyclyl, or a bond; and R.sub.6
is substituted or unsubstituted aryl, heteroaryl, cycloalkyl,
heterocycle, heterocycloalkyl, aryloxide, heteroaryl N-oxide, or
arylsulfide; provided when Y is a bond, then either R.sub.6 is
cycloalkyl, or R.sub.2 is alkylalkoxy or alkylthioalkoxy.
2. The compound of claim 1, wherein R.dbd.--CR.sub.1X'X'', X' is H
or OH, X'' is H, and R.sub.1 is aryl or substituted aryl.
3. The compound of claim 1, wherein R.sub.3 is H or t-butyl.
4. The compound of claim 1, wherein Y is substituted or
unsubstituted cycloalkyl, cycloalkenyl, or heterocyclyl.
5. The compound of claim 1, wherein Y is --CH.sub.2--CH.sub.2--;
CH.sub.3--CH<; --CH(CH.sub.3)--; --C(CH.sub.3).sub.2CH.sub.2--;
CH.sub.3--CH(phenyl)<; >CH(phenyl); or --CH.dbd.CH--.
6. The compound of claim 2, wherein R.sub.1 is
3,5-difluorophenyl.
7. The compound of claim 3, wherein R.sub.2 is methyl.
8. The compound of claim 1, selected from the group consisting of:
N-[2-tert-butyl-5-(1-methyl-4-phenylpiperidin-4-yl)-2H-pyrazol-3-yl]-2-[2-
-(3,5-difluorophenyl)-2-hydroxyacetylamino]propionamide,
2-[2-(3,5-difluorophenyl)-2-hydroxyacetylamino]-N-[5-(1-methyl-4-phenylpi-
peridin-4-yl)-2H-pyrazol-3-yl]propionamide,
2-[2-(3,5-difluoro-phenyl)-2-hydroxy-acetylamino]-N-[5-(1-phenyl-cyclopro-
pyl)-2H-pyrazol-3-yl]-propionamide,
2-[2-(3,5-difluoro-phenyl)-2-hydroxy-acetylamino]-N-[5-(1-phenyl-cyclopen-
tyl)-2H-pyrazol-3-yl]-propionamide,
2-[2-(3,5-Difluoro-phenyl)-2-hydroxy-acetylamino]-N-{5-[1-(4-fluoro-pheny-
l)-cyclopentyl]-2H-pyrazol-3-yl}-propionamide,
2-[2-(3,5-Difluoro-phenyl)-2-hydroxy-acetylamino]-N-{5-[1-(4-chloro-pheny-
l)-cyclopentyl]-2H-pyrazol-3-yl}-propionamide,
2-[2-(3,5-Difluoro-phenyl)-2-hydroxy-acetylamino]-N-[5-(1-phenyl-cyclohex-
yl)-2H-pyrazol-3-yl]-propionamide,
2-[2-(3,5-Difluoro-phenyl)-2-hydroxy-acetylamino]-N-{5-[1-(2-fluoro-pheny-
l)-cyclopentyl]-2H-pyrazol-3-yl}-propionamide,
2-[2-(3,5-Difluoro-phenyl)-2-hydroxy-acetylamino]-N-[5-(4-phenyl-tetrahyd-
ro-pyran-4-yl)-2H-pyrazol-3-yl]-propionamide,
N-[2-tert-Butyl-5-(4-phenyl-tetrahydro-pyran-4-yl)-2H-pyrazol-3-yl]-2-[2--
(3,5-difluoro-phenyl)-2-hydroxy-acetylamino]-propionamide,
N-(5-Cyclopentyl-2H-pyrazol-3-yl)-2-[2-(3,5-difluror-phenyl)2-2hydroxy-ac-
etylamino]-propionamide, and
N-[5-(1-Cyclopropyl-4-phenylpiperidin-4-yl)-2H-pyrazol-3-yl]-2-[2'-(3,5-d-
ifluorophenyl)-2'-hydroxylacetylamino]-propionamide.
9. A method for inhibiting .beta.-amyloid peptide release or
synthesis in a cell comprising administering to said cell a
compound according to claim 1, in an amount effective in inhibiting
the cellular release and/or synthesis of .beta.-amyloid
peptide.
10. A method for inhibiting .gamma.-secretase activity comprising
administering to a host an effective amount of the compound
according to claim 1.
11. A method for treating or preventing a neurological disorder
associated with .beta.-amyloid peptide production comprising
administering to a host a pharmaceutical formulation comprising a
therapeutically effective amount of the compound according to claim
1.
12. The method according to claim 11, wherein said neurological
disorder is Alzheimer's disease.
Description
[0001] This application is a divisional of U.S. application Ser.
No. 10/559,823, filed Mar. 1, 2007; which is National Stage of
International Application PCT/US2004/018202, filed Jun. 4, 2004,
published Feb. 3, 2005, under PCT Article 21(2) in English; which
claims the priority of U.S. Provisional Application 60/476,369,
filed Jun. 5, 2003. The contents of the above applications are
incorporated herein by reference in their entirety.
FIELD OF THE INVENTION
[0002] This invention relates to acylated amino acid amidyl
pyrazoles and related compounds that inhibit .beta.-amyloid peptide
release and/or its synthesis, and, accordingly, have utility in
treating Alzheimer's disease.
BACKGROUND OF THE INVENTION
[0003] Alzheimer's Disease (AD) is a degenerative brain disorder
characterized clinically by progressive loss of memory, cognition,
reasoning, judgment and emotional stability that gradually leads to
profound mental deterioration and ultimately death. AD is a very
common cause of progressive mental failure (dementia) in aged
humans and is believed to represent the fourth most common medical
cause of death in the United States. AD has been observed in races
and ethnic groups worldwide and presents a major present and future
public health problem. The disease is currently estimated to affect
about two to three million individuals in the United States alone.
AD is at present incurable. No treatment that effectively prevents
AD or reverses its symptoms and course is currently known.
[0004] The brains of individuals with AD exhibit characteristic
lesions termed senile (or amyloid) plaques, amyloid angiopathy
(amyloid deposits in blood vessels) and neurofibrillary tangles.
Large numbers of these lesions, particularly amyloid plaques and
neurofibrillary tangles, are generally found in several areas of
the human brain important for memory and cognitive function in
patients with AD. Smaller numbers of these lesions in a more
restrictive anatomical distribution are also found in the brains of
most aged humans who do not have clinical AD. Amyloid plaques and
amyloid angiopathy also characterize the brains of individuals with
Trisomy 21 (Down's Syndrome) and Hereditary Cerebral Hemorrhage
with Amyloidosis of the Dutch Type (HCHWA-D). At present, a
definitive diagnosis of AD usually requires observing the
aforementioned lesions in the brain tissue of patients who have
died with the disease or, rarely, in small biopsied samples of
brain tissue taken during an invasive neurosurgical procedure.
[0005] The principal chemical constituent of the amyloid plaques
and vascular amyloid deposits (amyloid angiopathy) characteristic
of AD and the other disorders mentioned above is an approximately
4.2 kilodalton (kD) protein of about 39-43 amino acids designated
the .beta.-amyloid peptide (.beta.AP) or sometimes A.beta.,
A.beta.P or .beta./A4. .beta.-Amyloid peptide was first purified
and a partial amino acid sequence was provided by Glenner, et al.
(Biochem. Biophys. Res. Commun., 120: 885-890)1984)).
.beta.-amyloid peptide is a small fragment of a much larger
precursor protein (APP), normally produced by cells in tissues of
various animals.
[0006] A.beta. is derived from cleavage of APP by protease systems,
collectively termed secretases. APP is first cleaved by .beta.
secretase to yield a .beta. stub, which is then cleaved by .gamma.
secretase to yield a .beta.-amyloid fragment that is secreted.
.beta. secretase generates the N-terminus of A.beta.. .gamma.
secretase generates C-terminal fragments ending at position 38, 39,
40, 42, and 43 or generating C-terminal extended precursors that
are subsequently truncated to the above polypeptides.
[0007] U.S. Pat. No. 6,153,652 discloses
N-(aryl/heteroaryl/alkyacetyl) amino acid amides, which inhibit
.beta. amyloid peptide release and/or its synthesis, and methods
for treating Alzheimer's disease with such compounds. U.S. Pat.
Nos. 6,191,166 and 6,211,235 each discloses a class of compounds,
which inhibit .beta. amyloid peptide release and/or its synthesis,
and methods for treating Alzheimer's disease with such compounds.
WO 00/38618 discloses succinoylaminobenzodiazepines and related
structures and methods for inhibiting .gamma.-secretase activity.
WO 00/77030 discloses statine-derived tetrapeptide inhibitors of
beta-secretase. WO 99/66934 discloses certain cyclic amino acid
compounds that inhibit .beta.-amyloid peptide release and/or its
synthesis and methods for treating Alzheimer's disease with such
compounds.
[0008] Despite the progress which has been made in understanding
the underlying mechanisms of AD and other .beta.-amyloid peptide
related diseases, there remains a need to develop methods and
compositions for treatment of the disease. The treatment methods
could be based on drugs that are capable of inhibiting
.beta.-amyloid peptide release and/or its synthesis in vivo.
Methods of treatment could target the formation of A.beta. through
the enzymes involved in the proteolytic processing of
.beta.-amyloid precursor protein. Compounds that inhibit
.gamma.-secretase activity, either directly or indirectly, control
the production of AP. Such inhibition of .gamma. secretase could
thereby reduce production of A.beta., which, thereby, reduces or
prevents the neurological disorders associated with .beta.-amyloid
peptide.
SUMMARY OF THE INVENTION
[0009] This invention is directed to a class of compounds that
inhibit .beta.-amyloid peptide release and/or its synthesis. The
class of compounds having the described properties are defined by
formula I below:
##STR00001##
[0010] The present invention is also directed to a pharmaceutical
formation comprising such compound in a pharmaceutically acceptable
salt form or prodrug thereof.
[0011] The present invention is directed to a method for inhibiting
.beta.-amyloid peptide release and/or synthesis and a method for
inhibiting .gamma.-secretase activity. The present invention is
also directed to a method for treating neurological disorders
associated with .beta.-amyloid peptide production. The method
comprises the steps of administering to a host a pharmaceutical
formulation comprising an effective amount of a compound of Formula
I. The compounds of Formula I are useful in the prevention of AD in
patients susceptible to AD and/or in the treatment of patients with
AD.
DETAILED DESCRIPTION OF THE INVENTION
[0012] This invention is directed to compounds that inhibit
.beta.-amyloid peptide release and/or its synthesis.
[0013] The class of compounds having the described properties is
defined by Formula I below:
##STR00002## [0014] or a pharmaceutically acceptable salt
therereof, [0015] wherein R is substituted or unsubstituted aryl,
cycloalkyl, heterocyclic, alkoxy, cycloalkoxy, aryloxy,
heteroaryloxy, alkylamino, cycloaklylamino, arylamino,
heteroarylamino; or [0016] R is
[0016] ##STR00003## [0017] wherein X' and X'' are each
independently hydrogen, hydroxy or fluoro, provided when one of X'
and X'' is fluoro, the other is not hydroxy; or
[0018] X' and X'' together form an oxo group,
[0019] Z is selected from the group consisting of alkyl, nitrogen,
oxygen, sulfur and a bond covalently linking R.sub.1 to
--CX'X''--
[0020] R.sub.1 is selected from the group consisting of hydrogen,
substituted or unsubstituted alkyl, alkenyl, aryl, cycloalkyl,
cycloalkenyl, heteroaryl, and heterocyclic;
[0021] R.sub.2 is selected from the group consisting of hydrogen,
C.sub.1-C.sub.4 alkyl, alkylalkoxy, alkylthioalkoxy, --COOR.sub.2a,
and --COR.sub.2a wherein R.sub.2a is hydrogen, C.sub.1-4 alkyl
(such as ethyl), cycloalkyl, or heterocycle (such as
pyrrolidinyl);
[0022] R.sub.3 is H, substituted or unsubstituted, linear-,
branched- or cyclo-alkyl or substituted or unsubustituted
phenyl;
[0023] R.sub.5 is --Y--R.sub.6, wherein Y is substituted or
unsubstituted alkyl, alkenyl, aryl, cycloalkyl, cycloalkenyl,
heteroaryl, heterocyclic, or a bond; and
[0024] R.sub.6 is substituted or unsubstituted aryl, heteroaryl,
cycloalkyl, heterocycloalkyl, aryloxide, heteroaryl N-oxide, or
arylsulfide;
[0025] provided when Y is a bond, then either R.sub.6 is
cycloalkyl, or R.sub.2 is alkylalkoxy or alkylthioalkoxy.
[0026] Examples of R.sub.5 include phenylmethyl, benzhydryl,
1-phenylcyclopropyl, 1-phenylcyclopentyl, 1-phenylcyclohexyl,
styryl, 1,1-dimethyl-1-phenylethyl, 1,1-dimethylbutyl,
2,2-dimethyl-2-phenylethyl, 4-phenyltetrahydropyran-4-yl,
1-phenylethyl, 3-phenylpropyl, 1-methyl-1-(phenylthio)ethyl,
1-cyclohexyl-1-methylethyl, 1-methyl-4-phenylpiperidin-4-yl,
1-cyclopropyl-4-phenylpiperidin-4-yl.
[0027] A preferred R.sub.2 is methyl. A preferred R.sub.3 is H or
t-butyl. A preferred R is (R.sub.1 Z)CX'X'', wherein said X' is H
or OH, X'' is H, Z is a bond, and R.sub.1 is alkyl, substituted
alkyl, aryl, or substituted aryl such as 3,5-difluorophenyl.
[0028] Preferred compounds of Formula I are shown as Formula
II:
##STR00004##
[0029] wherein R.sub.1 is alkyl, substituted alkyl, aryl, or
substituted aryl; X' is H or OH; R.sub.2 is CH.sub.3; R.sub.3 is H,
or t-butyl (with H being more preferred); Q is carbon; R.sub.7 is
aryl, substituted aryl, or U-Aryl, wherein U is O or CH.sub.2; and
R.sub.8 and R.sub.9 are independently H, or alkyl (such as
CH.sub.3), or R.sub.8, R.sub.9 and Q taken together form a
cycloalkyl or heterocycloalkyl.
Definition
[0030] The term ".beta.-amyloid peptide" refers to a 39-43 amino
acid peptide having a molecular weight of about 4.2 kD, which
peptide is substantially homologous to the form of the protein
described by Glenner, et al, (Biochem. Biophys. Res. Commun.,
120:885-890 (1984)) including mutations and post-translational
modifications of the normal .beta.-amyloid peptide. .beta.-amyloid
peptide is approximately a 39-43 amino acid fragment of a large
membrane-spanning glycoprotein, referred to as the .beta.-amyloid
precursor protein (APP). Its 43-amino acid sequence is disclosed in
U.S. Pat. No. 6,153,652.
[0031] "Alkyl" refers to monovalent alkyl groups preferably having
from 1 to 10 carbon atoms and more preferably 1 to 6 carbon atoms.
This term is exemplified by groups such as methyl, ethyl, n-propyl,
iso-propyl, n-butyl, iso-butyl, n-hexyl, and the like. The use of
the notation "C" followed by a numerical range preceding a defined
term, indicates a range of atoms intended to add further to the
definition. e.g., (C.sub.1-6)alkyl defines an alkyl group having
from 1 to 6 (inclusive) carbon atoms.
[0032] Unless otherwise constrained by a limitation of the alkyl
group, alkyl can optionally be substituted with from 1 to 3
substituents selected from the group consisting of hydroxy,
(C.sub.1-3) alkoxy, (C.sub.1-3)alkylthioxy, halo, acyl, acyloxy,
amino, aminoacyl, acylamino, alkoxycarbonyl, carboxyl, cyano,
phenyl optionally substituted with 1 to 2 halo atoms and
trifluoromethyl.
[0033] "Alkoxy" refers to the group "alkyl-O--" where alkyl is as
defined herein. Preferred alkoxy groups include, by way of example,
methoxy, ethoxy, n-propoxy, iso-propoxy, n-butoxy, tert-butoxy,
sec-butoxy, n-pentoxy, n-hexoxy, 1,2-dimethylbutoxy, and the
like.
[0034] "Alkylalkoxy" refers to the group "-alkylene-O-alkyl,"
substituted or unsubstituted, which includes by way of example,
methylenemethoxy (--CH.sub.2OCH.sub.3), ethylenemethoxy
(--CH.sub.2CH.sub.2OCH.sub.3), methylene-iso-propoxy
(--CH.sub.2--O--CH(CH.sub.3).sub.2) and the like.
[0035] "Alkylthioalkoxy" refers to the group "-alkylene-S-alkyl,"
substituted or unsubstituted, which includes by way of example,
methylenethiomethoxy (--CH.sub.2SCH.sub.3), ethylenethiomethoxy
(--CH.sub.2CH.sub.2SCH.sub.3), methylene-iso-thiopropoxy
(--CH.sub.2--S--CH(CH.sub.3).sub.2) and the like.
[0036] "Alkenyl" refers to alkenyl groups preferably having from 2
to 10 carbon atoms and more preferably 2 to 6 carbon atoms and
having at least 1 and preferably from 1-2 sites of alkenyl
unsaturation. Preferred alkenyl groups include ethenyl
(--CH.dbd.CH.sub.2), allyl or n-2 propenyl
(--CH.sub.2CH.dbd.CH.sub.2), iso-propenyl
(--C(CH.sub.3).dbd.CH.sub.2), 2-butenyl
(--CH.sub.2CH.dbd.CHCH.sub.3) and the like.
[0037] "Alkynyl" refers to alkynyl groups preferably having from 2
to 10 carbon atoms and more preferably 2 to 6 carbon atoms and
having at least 1 and preferably from 1-2 sites of alkynyl
unsaturation. Preferred alkynyl groups include ethynyl
(--C.ident.CH), propargyl (--CH.sub.2--C.ident.CH) and the
like.
[0038] "Acyl" refers to the groups alkyl-C(O)--, aryl-C(O)--, and
heteroaryl-C(O)-- where alkyl, aryl and heteroaryl are as defined
herein.
[0039] "Acylamino" refers to the group --C(O)NRR where each R is
independently hydrogen or alkyl where alkyl is as defined
herein.
[0040] "Aminoacyl" refers to the group --NRC(O)R where each R is
independently hydrogen or alkyl where alkyl is as defined
herein.
[0041] "Acyloxy" refers to the groups alkyl-C(O)O--, aryl-C(O)O--,
heteroaryl-C(O)O--, and heterocyclic-C(O)O-- where alkyl, aryl,
heteroaryl and heterocyclic are as defined herein.
[0042] "Aryl" refers to an unsaturated aromatic carbocyclic group
of from 6 to 14 carbon atoms having a single ring (e.g., phenyl) or
multiple condensed rings (e.g., naphthyl or anthryl). Preferred
aryls include phenyl, naphthyl and the like.
[0043] "Arylalkyl" refers to aryl-alkyl- groups preferably having
from 1 to 6 carbon atoms in the alkyl moiety and from 6 to 10
carbon atoms in the aryl moiety. Such arylalkyl groups are
exemplified by benzyl, phenethyl and the like.
[0044] "Arylalkenyl" refers to aryl-alkenyl- groups preferably
having from 1 to 6 carbon atoms in the alkenyl moiety and from 6 to
10 carbon atoms in the aryl moiety.
[0045] "Arylalkynyl" refers to aryl-alkynyl- groups preferably
having from 1 to 6 carbon atoms in the alkynyl moiety and from 6 to
10 carbon atoms in the aryl moiety.
[0046] "Aryloxy" refers to the group aryl-O-- wherein the aryl
group is as defined above including optionally substituted aryl
groups as also defined above.
[0047] The terms "amide" and "amido" refer to a functional group
containing a carbon atom double-bonded to an oxygen atom and
additionally singly bonded to a nitrogen atom [--C(O)--N].
"Primary" amide describes an unsubstituted amide group
[--C(O)--NH.sub.2]. "Secondary" and "tertiary" amides are amides in
which nitrogen is substituted with one and two non-hydrogen groups
respectively.
[0048] "Cycloalkyl" refers to cyclic alkyl groups of from 3 to 10
carbon atoms having a single cyclic ring or multiple condensed
rings which can be optionally substituted with from 1 to 3 alkyl
groups. Such cycloalkyl groups include, by way of example, single
ring structures such as cyclopropyl, cyclobutyl, cyclopentyl,
cyclooctyl, 1-methylcyclopropyl, 2-methylcyclopentyl,
2-methylcyclooctyl, and the like, or multiple ring structures such
as adamantyl, and the like.
[0049] "Cycloalkenyl" refers to cyclic alkenyl groups of from 4 to
8 carbon atoms having a single cyclic ring and at least one point
of internal unsaturation which can be optionally substituted with
from 1 to 3 alkyl groups. Examples of suitable cycloalkenyl groups
include, for instance, cyclobut-2-enyl, cyclopent-3-enyl,
cyclooct-3-enyl and the like.
[0050] "Halo" or "halogen" refers to fluoro, chloro, bromo and iodo
and preferably is either chloro or fluoro.
[0051] "Heteroaryl" refers to a monovalent aromatic carbocyclic
group of from 1 to 10 carbon atoms and 1 to 4 heteroatoms selected
from oxygen, nitrogen and sulfur within the ring. Such heteroaryl
groups can have a single ring (e.g., pyridyl or furyl) or multiple
condensed rings (e.g., indolizinyl or benzothienyl).
[0052] "Heterocycle" or "heterocyclic" refers to a saturated or
unsaturated group having a single ring or multiple condensed rings,
from 1 to 8 carbon atoms and from 1 to 4 hetero atoms selected from
nitrogen, sulfur or oxygen within the ring. Such heterocyclic
groups can have a single ring (e.g., piperidinyl or
tetrahydrofuryl) or multiple condensed rings (e.g., indolinyl,
dihydrobenzofuran or quinuclidinyl). Preferred heterocycles include
piperidinyl, pyrrolidinyl and tetrahydrofuryl.
[0053] Examples of heterocycles and heteroaryls include, but are
not limited to, furan, thiophene, thiazole, oxazole, pyrrole,
imidazole, pyrazole, pyridine, pyrazine, pyrimidine, pyridazine,
indolizine, isoindole, indole, indazole, purine, quinolizine,
isoquinoline, quinoline, phthalazine, naphthylpyridine,
quinoxaline, quinazoline, cinnoline, pteridine, carbazole,
carboline, phenanthridine, acridine, phenanthroline, isothiazole,
phenazine, isoxazole, phenoxazine, phenothiazine, imidazolidine,
imidazoline, piperidine, piperazine, pyrrolidine, indoline and the
like.
[0054] "Thiol" refers to the group --SH.
[0055] "Thioalkoxy" refers to the group --S-alkyl.
[0056] "Thioaryloxy" refers to the group aryl-S-- wherein the aryl
group is as defined above including optionally substituted aryl
groups as also defined above.
[0057] Unless otherwise constrained by the definition for the
individual substituent, aryl, cycloalkyl, heteroaryl, and
heterocyclic groups can be optionally substituted with 1 to 3
substituents selected from the group consisting of alkyl, alkenyl,
aryl, cycloalkyl, cycloalkenyl, heteroaryl, heterocyclic, alkaryl,
alkcycloalkyl, alkcycloalkenyl, alkheteroaryl, alkheterocyclic,
alkoxy, aryloxy, halo, nitro, hydroxy, amino, acyl, acyloxy,
aminoacyl, acylamino, carboxy, cyano, alkoxycarbonyl, thioalkoxy,
thioaryloxy and the like. Preferred substituents include alkyl,
alkoxy, halo, cyano, nitro, hydroxy, trihalomethyl, thioalkoxy, and
amino.
[0058] "Pharmaceutically acceptable salt" refers to
pharmaceutically acceptable salts of a compound of Formula I which
salts are derived from a variety of organic and inorganic counter
ions well known in the art and include, by way of example only,
lithium, sodium, potassium, calcium, magnesium, ammonium,
tetraalkylammonium, and the like; and when the molecule contains a
basic functionality, salts of organic or inorganic acids, such as
hydrochloride, hydrobromide, tartrate, mesylate, acetate, maleate,
oxalate and the like.
Pharmaceutical Formulations
[0059] When employed as pharmaceuticals, the compounds of Formula I
are usually administered in the form of pharmaceutical
compositions. These compounds can be administered by a variety of
routes including oral, rectal, transdermal, subcutaneous,
intravenous, intramuscular, and intranasal. These compounds are
effective as both injectable and oral compositions. Such
compositions are prepared in a manner well known in the
pharmaceutical art and comprise at least one active compound.
[0060] This invention also includes pharmaceutical compositions
that contain, as the active ingredient, one or more of the
compounds of Formula I, or a pharmaceutically accepted form or
prodrug thereof, associated with pharmaceutically acceptable
carriers. In making the compositions of this invention, the active
ingredient is usually mixed with an excipient, diluted by an
excipient or enclosed within such a carrier which can be in the
form of a capsule, sachet, paper or other container. When the
excipient serves as a diluent, it can be a solid, semi-solid, or
liquid material, which acts as a vehicle, carrier or medium for the
active ingredient. Thus, the compositions can be in the form of
tablets, pills, powders, lozenges, sachets, cachets, elixirs,
suspensions, emulsions, solutions, syrups, aerosols (as a solid or
in a liquid medium), ointments containing, for example, up to 10%
by weight of the active compound, soft and hard gelatin capsules,
suppositories, sterile injectable solutions, and sterile packaged
powders.
[0061] In preparing a formulation, it may be necessary to mill the
active compound to provide the appropriate particle size prior to
combining with the other ingredients. If the active compound is
substantially insoluble, it ordinarily is milled to a particle size
of less than 200 mesh. If the active compound is substantially
water soluble, the particle size is normally adjusted by milling to
provide a substantially uniform distribution in the formulation,
e.g. about 40 mesh.
[0062] Some examples of suitable excipients include lactose,
dextrose, sucrose, sorbitol, mannitol, starches, gum acacia,
calcium phosphate, alginates, tragacanth, gelatin, calcium
silicate, microcrystalline cellulose, polyvinylpyrrolidone,
cellulose, sterile water, syrup, and methyl cellulose. The
formulations can additionally include: lubricating agents such as
talc, magnesium stearate, and mineral oil; wetting agents;
emulsifying and suspending agents; preserving agents such as
methyl- and propylhydroxy-benzoates; sweetening agents; and
flavoring agents. The compositions of the invention can be
formulated so as to provide quick, sustained or delayed release of
the active ingredient after administration to the patient by
employing procedures known in the art.
[0063] The compositions are preferably formulated in a unit dosage
form, each dosage containing from about 1 to about 100 mg, more
usually about 10 to about 30 mg, of the active ingredient. The term
"unit dosage forms" refers to physically discrete units suitable as
unitary dosages for human subjects and other mammals, each unit
containing a predetermined quantity of active material calculated
to produce the desired therapeutic effect, in association with a
suitable pharmaceutical excipient. Preferably, the compound of
formula I above is employed at no more than about 20 weight percent
of the pharmaceutical composition, more preferably no more than
about 15 weight percent, with the balance being pharmaceutically
inert carriers.
[0064] The active compound is effective over a wide dosage range
and is generally administered in a pharmaceutically effective
amount. The amount of the compound actually administered will be
determined by a physician, in view of the relevant circumstances,
including the condition to be treated, the chosen route of
administration, the actual compound administered, the age, weight,
and response of the individual patient, the severity of the
patient's symptoms, and the like.
[0065] For preparing solid compositions such as tablets, the
principal active ingredient is mixed with a pharmaceutical
excipient to form a solid preformulation composition containing a
homogeneous mixture of a compound of the present invention. When
referring to these preformulation compositions as homogeneous, it
is meant that the active ingredient is dispersed evenly throughout
the composition so that the composition can be readily subdivided
into equally effective unit dosage forms such as tablets, pills and
capsules. This solid preformulation is then subdivided into unit
dosage forms of the type described above containing from, for
example, 0.1 to about 500 mg of the active ingredient of the
present invention.
[0066] The tablets or pills of the present invention can be coated
or otherwise compounded to provide a dosage form affording the
advantage of prolonged action. For example, the tablet or pill can
comprise an inner dosage and an outer dosage component, the latter
being in the form of an envelope over the former. The two
components can be separated by an enteric layer which serves to
resist disintegration in the stomach and permit the inner component
to pass intact into the duodenum or to be delayed in release. A
variety of materials can be used for such enteric layers or
coatings, such materials including a number of polymeric acids and
mixtures of polymeric acids with such materials as shellac, cetyl
alcohol, and cellulose acetate.
[0067] The liquid forms in which the novel compositions of the
present invention can be incorporated for administration orally or
by injection include aqueous solutions, suitably flavored syrups,
aqueous or oil suspensions, and flavored emulsions with edible oils
such as corn oil, cottonseed oil, sesame oil, coconut oil, or
peanut oil, as well as elixirs and similar pharmaceutical
vehicles.
[0068] Compositions for inhalation or insufflation include
solutions and suspensions in pharmaceutically acceptable, aqueous
or organic solvents, or mixtures thereof, and powders. The liquid
or solid compositions may contain suitable pharmaceutically
acceptable excipients as described supra. Preferably the
compositions are administered by the oral or nasal respiratory
route for local or systemic effect. Compositions in preferably
pharmaceutically acceptable solvents may be nebulized by use of
inert gases. Nebulized solutions may be inhaled directly from the
nebulizing device or the nebulizing device may be attached to a
face mask tent, or intermittent positive pressure breathing
machine. Solution, suspension, or powder compositions may be
administered, preferably orally or nasally, from devices which
deliver the formulation in an appropriate manner.
[0069] Another preferred formulation employed in the methods of the
present invention employs transdermal delivery devices ("patches").
Such transdermal patches may be used to provide continuous or
discontinuous infusion of the compounds of the present invention in
controlled amounts. The construction and use of transdermal patches
for the delivery of pharmaceutical agents is well known in the art.
See. e.g., U.S. Pat. No. 5,023,252, herein incorporated by
reference. Such patches may be constructed for continuous,
pulsatile, or on demand delivery of pharmaceutical agents.
[0070] Often, it is desirable or necessary to introduce the
pharmaceutical composition to the brain, either directly or
indirectly. Direct techniques usually involve placement of a drug
delivery catheter into the host's ventricular system to bypass the
blood-brain barrier. One such implantable delivery system used for
the transport of biological factors to specific anatomical regions
of the body is described in U.S. Pat. No. 5,011,472 which is herein
incorporated by reference.
[0071] Indirect techniques, which are generally preferred, usually
involve formulating the compositions to provide for drug
latentiation by the conversion of hydrophilic drugs into
lipid-soluble drugs. Latentiation is generally achieved through
blocking of the hydroxy, carbonyl, sulfate, and primary amine
groups present on the drug to render the drug more lipid soluble
and amenable to transportation across the blood-brain barrier.
Alternatively, the delivery of hydrophilic drugs may be enhanced by
intra-arterial infusion of hypertonic solutions which can
transiently open the blood-brain barrier.
[0072] Other suitable formulations for use in the present invention
can be found in Remington's Pharmaceutical Sciences, Mace
Publishing Company, Philadelphia, Pa., 17th ed. (1985).
Utility
[0073] The present invention is directed to a method for inhibiting
.beta.-amyloid peptide release and/or synthesis in a cell and a
method for inhibiting .gamma.-secretase activity. The invention is
also directed to a method for preventing or treating of
neurological disorders associated with .beta.-amyloid peptide
production. The method comprises the steps of administering to a
host in need of such treatment a pharmaceutical formulation
comprising a therapeutically effective amount of a compound of
Formula I. The compounds of Formula I are useful in the prevention
of AD in patients susceptible to AD and/or in the treatment of
patients with AD.
[0074] A.beta. production has been implicated in the pathology of
Alzheimer's Disease (AD). The compounds of the present invention
have utility for the prevention and treatment of AD by inhibiting
A.beta. production. Methods of treatment target formation of
A.beta. production through the enzymes involved in the proteolytic
processing of .beta.-amyloid precursor protein. Compounds that
inhibit .gamma. secretase activity, either directly or indirectly,
control the production of A.beta.. Such inhibition of .gamma.
secretase reduces production of A.beta., and is expected to reduce
or prevent the neurological disorders associated with A.beta..sub.\
such as Alzheimer's Disease.
[0075] Compounds of Formula I are expected to possess
.gamma.-secretase inhibitory activity or inhibit A.beta.
production. Cellular screening methods for inhibitors of A.beta.
production, testing methods for the in vivo suppression of A.beta.
production, and assays for the detection of secretase activity are
known in the art and have been disclosed in many publications,
including WO 98/22493 and WO 01/19797, EP 0652009, U.S. Pat. Nos.
5,703,129 5,593,846; 6,211,235 and 6,207,710, all hereby
incorporated by reference.
[0076] Compounds provided by this invention are useful as standards
and reagents in determining the ability of a potential
pharmaceutical reagent to inhibit A.beta. production. These can be
provided in a kit comprising a compound of this invention.
[0077] A compound is considered to be active if it has an IC.sub.50
or K.sub.i value of less than about 100 .mu.M for the inhibition of
A.beta. production.
Synthesis and Preparation of Compounds Having Formula I
[0078] The compounds of the present invention can be prepared in a
number of ways known to one skilled in the art of organic
synthesis. The compounds of the present invention can be
synthesized using the methods described below, together with
synthetic methods known in the art of synthetic organic chemistry,
or variations thereon as appreciated by those skilled in the art.
Preferred methods include, but are not limited to, those described
below. All references cited herein are hereby incorporated in their
entirety by reference.
[0079] The novel compounds of this invention may be prepared using
the reactions and techniques described in this section. The
reactions are performed in solvents appropriate to the reagents and
materials employed and are suitable for the transformations being
effected. Also, in the description of the synthetic methods
described below, it is to be understood that all proposed reaction
conditions, including choice of solvent, reaction atmosphere,
reaction temperature, duration of the experiment and workup
procedures, are chosen to be the conditions standard for that
reaction, which should be readily recognized by one skilled in the
art. It is understood by one skilled in the art of organic
synthesis that the functionality present on various portions of the
molecule must be compatible with the reagents and reactions
proposed. Such restrictions to the substituents which are
compatible with the reaction conditions will be readily apparent to
one skilled in the art and alternate methods must then be used.
Construction of Acylated Amino Acid Amidyl Pyrazoles.
[0080] A variety of substituted acylated amino acid amidyl
pyrazoles can be prepared using the corresponding carboxylic acids
in this fashion. The following Methods A-f show general methods for
preparing acylated amino acid amidyl pyrazoles.
Synthesis of Pyrazoles (Type 8 and 9).
##STR00005##
[0082] A variety of 5' substituted pyrazoles can be prepared using
carboxylic acid (1), amino acids (4, natural and unatural) and an
additional carboxylic acid (7). In some instances, protecting group
strategies on R, R.sub.2 and/or R.sub.5 are required, the methods
described in Protective Groups In Organic Synthesis (Theodora W.
Greene and Peter G. M. Wuts, Wiley and Sons) were employed to
address these cases.
##STR00006##
[0083] Method A
[0084] In a dried round bottom flask, a solution of carboxylic acid
1 is prepared. To the reaction solution, N,N'-carbonyldiimidazole
is added. In a separate round bottom flask, a solution of
cyanoacetic acid is added with isopropylmagnesium chloride. The two
reaction mixtures are mixed together and after a time sufficient
for reaction, combined with acetic acid to bring to .about.pH 5.
The crude material is further processed to give a yellowish oil as
ketonitrile product 2.
##STR00007##
[0085] Method B
[0086] A solution of ketonitrile 2, tert-butylhydrazine, and
triethylamine in absolute ethanol is refluxed for a time sufficient
for reaction to proceed. After cooling to room temperature, the
reaction solution is concentrated, extracted with EtOAc, and dried.
The residual crude material is flash chromatographed to afford the
t-butyl amino pyrazole 3.
##STR00008##
[0087] Method C
[0088] General Procedure for POCl.sub.3 Coupling of t-butyl amino
pyrazole 3 with amino acid 4. A solution of 3 and amino acid 4 in
suitable solvent is added with POCl.sub.3. The reaction mixture is
then extracted, dried, and the solvent removed. Purification of the
material affords product 5.
##STR00009##
[0089] Method D
[0090] A solution of N-tert-butyl protected pyrazole 5 is stirred
in a solution of methylene chloride/trifluoroacetic acid. The
volatiles are evaporated off to yield the corresponding salt of the
desired amine 6.
##STR00010##
[0091] Method E
[0092] The carboxylic acid 7, amine 6, triethylamine (TEA) and
hydroxybenzotriazole hydrate (HOBT) are mixed. The mixture is then
added with 1-ethyl-3-(3'-dimethylaminopropyl)carbodiimide
hydrochloride (EDCI). The reaction mixture is then purified to
yield the desired product 8.
##STR00011##
[0093] Method F
[0094] A solution of N-tert-butyl protected pyrazole 8 in formic
acid is refluxed then added to saturated NaHCO.sub.3(aq). The
quenched solution is extracted, dried, and vacuum filtered. The
crude material is chromatographed on silica to give the t-butyl
deprotected product 9. In the event that functionalities on R,
R.sub.2 or R.sub.5 have been formylated as a result of Method F,
the resulting formyl group is removed.
##STR00012##
Examples
Examples A-H Show Methods for Preparing Acylted Amino Acid Amidyl
Pyrazoles
##STR00013## ##STR00014##
[0095] Example A
[0096] 4-(2-Cyanoacetyl)-4-phenylpiperidine-1-carboxylic acid
tert-butyl ester. A solution of 2.52 g (30 mmol) of cyanoacetic
acid in 80 mL of dry THF was stirred at -78.degree. C. as 24.6 mL
(49.2 mmol) of 2.0 M i-PrMgCl in THF was added. After 1 h, a
solution of 3.00 g (10 mmol)
1-tert-butoxycarbonyl-4-phenylpiperidine-4-carboxylic acid
(Maybridge, cat. no. JFD 01929) and 1.93 g (12 mmol) of
1,1'-carbonyldiimidazole in 20 mL of THF was added. The reaction
mixture was allowed to warm to rt. After 16 h, the mixture was
poured into 300 mL of water and the pH lowered to 4 with conc.
AcOH. The mixture was then extracted with EtOAc, dried over
MgSO.sub.4 and concentrated. Purification on silica gel using 1:1
EtOAc-hexanes as eluant afforded
4-(2-cyanoacetyl)-4-phenylpiperidine-1-carboxylic acid tert-butyl
ester.
Example B
[0097]
4-(5-Amino-1-tert-butyl-1H-pyrazol-3-yl)-4-phenylpiperidine-1-carbo-
xylic acid tert-butyl ester. A solution of 1.66 g (5.0 mmol) of
4-(2-cyanoacetyl)-4-phenylpiperidine-1-carboxylic acid tert-butyl
ester, 2.20 mL (16 mmol) of Et.sub.3N and 2.00 g (16 mmol) of
tert-butylhydrazine hydrochloride in 20 mL of absolute EtOH was
heated to 100.degree. C. in a sealed tube. After 64 h, the reaction
mixture was cooled to rt and concentrated. Purification on silica
gel using 30:70 EtOAc-hexanes as eluant afforded
4-(5-amino-1-tert-butyl-1H-pyrazol-3-yl)-4-phenylpiperidine-1-carboxylic
acid tert-butyl ester.
Example C
[0098]
4-[5-(2-Benzyloxycarbonylamino-propionylamino)-1-tert-butyl-1H-pyra-
zol-3-yl]-4-phenylpiperidine-1-carboxylic acid tert-butyl ester. A
solution of 0.68 g (1.7 mmol) of
4-(5-amino-1-tert-butyl-1H-pyrazol-3-yl)-4-phenylpiperidine-1-carboxylic
acid tert-butyl ester and 0.39 g (1.7 mmol) of
carbobenzyloxy-L-alanine in 9.0 mL of dry pyridine was stirred at
-10.degree. C. as 0.18 mL (1.9 mmol) of POCl.sub.3 was added. After
25 min, the reaction mixture was poured into 1M HCl and extracted
with EtOAc. The organic layer was washed with sat. aq. NaHCO.sub.3,
dried over MgSO.sub.4 and concentrated. Purification on silica gel
using 50:50 EtOAc-hexanes as eluant afforded
4-[5-(2-benzyloxycarbonylamino-propionylamino)-1-tert-butyl-1H-pyrazol-3--
yl]-4-phenylpiperidine-1-carboxylic acid tert-butyl ester.
Example D
[0099]
{1-[2-tert-Butyl-5-(4-phenylpiperidin-4-yl)-2H-pyrazol-3-ylcarbamoy-
l]ethyl}carbamic acid benzyl ester. A solution of 0.54 g (0.9 mmol)
of
4-[5-(2-benzyloxycarbonylamino-propionylamino)-1-tert-butyl-1H-pyrazol-3--
yl]-4-phenylpiperidine-1-carboxylic acid tert-butyl ester in 4.0 mL
of neat trifluoroacetic acid was stirred at rt for 15 min. The
reaction mixture was then poured into sat. aq. NaHCO.sub.3 and
extracted with EtOAc. The organic layer was dried over MgSO.sub.4
and concentrated to afford
{1-[2-tert-butyl-5-(4-phenylpiperidin-4-yl)-2H-pyrazol-3-ylcarbamo-
yl]ethyl}-carbamic acid benzyl ester which was used without further
purification.
Example E
[0100]
{1-[2-tert-Butyl-5-(1-methyl-4-phenylpiperidin-4-yl)-2H-pyrazol-3-y-
lcarbamoyl]ethyl}carbamic acid benzyl ester. A solution of 0.67 g
(1.3 mmol) of
{1-[2-tert-butyl-5-(4-phenylpiperidin-4-yl)-2H-pyrazol-3-ylcarba-
moyl]ethyl}carbamic acid benzyl ester, 0.33 mL (4.0 mmol) of 37%
aq. formaldehyde, 0.33 g (4.0 mmol) of NaOAc and 0.27 g (4.3 mmol)
of NaCNBH.sub.3 in 10 mL of MeOH was stirred at rt. After 16 h, the
reaction mixture was concentrated and extracted with water and
EtOAc. The organic layer was dried over MgSO.sub.4, and
concentrated. Purification on silica gel using 4% Et.sub.3N-MeOH as
eluant afforded
{1-[2-tert-butyl-5-(1-methyl-4-phenylpiperidin-4-yl)-2H-pyrazol-3-ylcarba-
moyl]ethyl}carbamic acid benzyl ester.
Example F
[0101]
2-Amino-N-[2-tert-butyl-5-(1-methyl-4-phenylpiperidin-4-yl)-2H-pyra-
zol-3-yl]propionamide. A suspension of 0.36 g (0.7 mmol) of
{1-[2-tert-butyl-5-(1-methyl-4-phenylpiperidin-4-yl)-2H-pyrazol-3-ylcarba-
moyl]ethyl}carbamic acid benzyl ester and 0.36 g of 10% Pd/C in 5.0
mL of absolute EtOH was agitated under 30 psi H.sub.2 for 16 h. The
reaction mixture was filtered through Celite and 0.36 g of fresh
10% Pd/C was added to the filtrate which was again agitated under
30 psi H.sub.2 for 16 h. This process was repeated once more and,
after a final filtration, the filtrate was concentrated.
Purification on silica gel using 4% Et.sub.3N-MeOH as eluant
afforded
2-amino-N-[2-tert-butyl-5-(1-methyl-4-phenylpiperidin-4-yl)-2H-pyrazol-3--
yl]propionamide.
Example G
[0102]
N-[2-tert-Butyl-5-(1-methyl-4-phenylpiperidin-4-yl)-2H-pyrazol-3-yl-
]-2-[2-(3,5-difluorophenyl)-2-hydroxyacetylamino]propionamide. A
solution of 0.14 g (0.4 mmol) of
2-amino-N-[2-tert-butyl-5-(1-methyl-4-phenylpiperidin-4-yl)-2H-pyrazol-3--
yl]propionamide and 0.08 g (0.4 mmol) of (S)-3,5-difluoromandelic
acid (Garofalo, et al. Bioorg. Med. Chem, Lett, 2002, 12, 3051) in
5.0 mL of THF was stirred at rt as 0.06 g (0.4 mmol) of HOBT and
0.08 mL (0.7 mmol) of NMM was added followed by 0.90 g (0.5 mmol)
of EDC.cndot.HCl. After 16 h, the reaction was quenched with 1M HCl
and extracted with EtOAc. The organic layer was washed with sat.
aq. NaHCO.sub.3, dried over MgSO.sub.4 and concentrated.
Purification on silica gel using 4% Et.sub.3N-MeOH as eluant
afforded
N-[2-tert-butyl-5-(1-methyl-4-phenylpiperidin-4-yl)-2H-pyrazol-3-yl]-2-[2-
-(3,5-difluorophenyl)-2-hydroxyacetylamino]propionamide.
Example H
[0103]
2-[2-(3,5-Difluorophenyl)-2-hydroxyacetylamino]-N-[5-(1-methyl-4-ph-
enylpiperidin-4-yl)-2H-pyrazol-3-yl]propionamide. A solution of
0.04 g (0.1 mmol) of
N-[2-tert-butyl-5-(1-methyl-4-phenylpiperidin-4-yl)-2H-pyrazol-3-yl]-2-[2-
-(3,5-difluorophenyl)-2-hydroxyacetylamino]propionamide in 2.0 mL
of neat trifluoroacetic acid was refluxed for 15 min. The reaction
mixture was then cooled to rt, quenched with sat. aq. NaHCO.sub.3
and extracted with EtOAc. The organic layer was dried over
Na.sub.2SO.sub.4 and concentrated. Purification on silica gel
afforded
2-[2-(3,5-difluorophenyl)-2-hydroxyacetylamino]-N-[5-(1-methyl-4-phenylpi-
peridin-4-yl)-2H-pyrazol-3-yl]propionamide.
[0104] Methods A-E or A-F were utilized to prepared the following
compounds (Examples 1-107).
Example 1
##STR00015##
[0106] MS 415.0 (M+H), 436.8 (M+Na), 452.6 (M+K).
[0107] Anal Calcd for C.sub.21H.sub.20F.sub.2N.sub.4O.sub.3: C,
60.87; H, 4.86; N, 13.52. Found: C, 60.73; H, 4.73; N, 13.28.
[0108] .sup.1H NMR (CDCl.sub.3, 300 MHz) .delta. 11.97 (s, 1H),
7.99 (d, J=9.3 Hz, 1H), 7.21 (m, 5H), 7.09 (d, J=6.1 Hz, 2H), 6.79
(m, 1H), 6.37 (s, 1H), 5.11 (s, 1H), 4.74 (m, 1H), 3.85 (m, 2H),
1.35 (d, J=6.6 Hz, 3H).
Example 2
##STR00016##
[0110] MS 415.2 (M+H), 436.8 (M+Na).
[0111] Anal Calcd for
C.sub.21H.sub.20F.sub.2N.sub.4O.sub.3.1/4H.sub.2O: C, 60.21; H,
4.93; N, 13.57. Found: C, 60.24; H, 4.76; N, 13.31.
[0112] .sup.1H NMR (CDCl.sub.3, 300 MHz) .delta. 11.82 (s, 1H),
8.03 (d, J=9.3 Hz, 1H), 7.23 (m, 5H), 6.95 (d, J=6.0 Hz, 2H), 6.57
(m, 1H), 6.21 (s, 1H), 5.16 (s, 1H), 4.87 (m, 1H), 3.84 (m, 2H),
1.41 (d, J=6.6 Hz, 3H).
Example 3
##STR00017##
[0114] MS 440.8 (M+H), 463.2 (M+Na).
[0115] Anal Calcd for C.sub.23H.sub.22F.sub.2N.sub.4O.sub.3: C,
62.72; H, 5.03; N, 12.72. Found: C, 62.69; H, 5.03; N, 12.68.
[0116] .sup.1H NMR (CDCl.sub.3, 300 MHz) .delta. 11.55 (s, 1H),
11.33 (s(br), 1H), 7.70 (d, J=9.9 Hz, 1H), 7.23 (m, 5H), 7.04 (d,
J=7.1 Hz, 2H), 6.76 (t, J=8.2 Hz, 1H), 6.30 (s, 1H), 5.39 (s(br),
1H), 4.71 (m, 1H), 1.33 (m, 7H).
Example 4
##STR00018##
[0118] MS 469.3 (M+H), 491.3 (M+Na).
[0119] Anal Calcd for C.sub.25H.sub.26F.sub.2N.sub.4O.sub.3: C,
64.09; H, 5.59; N, 11.96. Found: C, 64.06; H, 5.52; N,11.66.
[0120] .sup.1H NMR (CDCl.sub.3, 300 MHz) .delta. 12.01 (s, 1H),
11.75 (s(br), 1H), 7.84 (d, J=9.3 Hz, 1H), 7.18 (m, 7H), 6.81 (t,
J=8.8 Hz, 1H), 6.43 (s, 1H), 5.95 (s(br), 1H), 5.14 (s, 1H), 4.73
(m, 1H), 2.35 (m, 2H), 2.14 (m, 2H), 1.65 (m, 4H), 1.41 (d, J=6.6
Hz, 3H).
Example 5
##STR00019##
[0122] MS 477.0 (M+H).
[0123] Anal Calcd for C.sub.23H.sub.23ClF.sub.2N.sub.4O.sub.3: C,
57.93; H, 4.86; N, 11.75. Found: C, 57.68; H, 4.92; N, 11.83.
[0124] .sup.1H NMR (CDCl.sub.3, 300 MHz) .delta. 11.75 (s(br), 1H),
7.75 (d, J=9.3 Hz, 1H), 7.16 (m, 4H), 7.02 (d, J=6.0 Hz, 2H), 6.77
(t, J=8.8 Hz, 1H), 6.50 (s, 1H), 5.08 (s, 1H), 4.74 (m, 1H), 1.64
(s, 6H), 1.43 (d, J=6.6 Hz, 3H).
Example 6
##STR00020##
[0126] MS 488.3 (M+H), 510.1 (M+Na).
[0127] Anal Calcd for
C.sub.23H.sub.23F.sub.2N.sub.5O.sub.5.H.sub.2O: C, 55.64; H, 4.87;
N, 14.11. Found: C, 55.60; H, 4.82; N, 13.88.
[0128] .sup.1H NMR (CDCl.sub.3, 300 MHz) .delta. 12.14 (s(br), 1H),
12.03 (s(br), 1H), 8.04 (d, J=8.9, 2H), 7.95 (d, J=8.2 Hz, 1H),
7.33 (d, J=8.8 Hz, 2H), 7.01 (d, J=6.0 Hz, 2H), 6.77 (m, 1H), 5.98
(s(br), 1H), 5.11 (s, 1H), 4.74 (m, 1H), 1.68 (s, 3H), 1.66 (s,
3H), 1.44 (d, J=6.6 Hz, 3H).
Example 7
##STR00021##
[0130] MS 505.3 (M+H), 527.1 (M+Na).
[0131] Anal Calcd for
C.sub.28H.sub.26F.sub.2N.sub.4O.sub.3.3/4H.sub.2O: C, 64.92; H,
5.35; N, 10.81. Found: C, 64.94; H, 5.24; N, 10.81.
[0132] .sup.1H NMR (CD.sub.3OD, 300 MHz) .delta. 7.24 (m, 6H), 7.09
(m, 6H), 6.83 (m, 1H), 6.13 (s, 1H), 5.04 (s, 1H), 4.46 (m, 1H),
2.10 (s, 3H), 1.36 (d, J=7.14, 3H).
Example 8
##STR00022##
[0134] MS 491.1 (M+H), 513.1 (M+Na).
[0135] Anal Calcd for C.sub.27H.sub.24F.sub.2N.sub.4O.sub.3: C,
66.11; H, 4.93; N, 11.42. Found: C, 65.79; H, 5.22; N, 11.25.
[0136] .sup.1H NMR (CDCl.sub.3, 300 MHz) .delta. 7.74 (d, J=9.3 Hz,
1H), 7.21 (m, 10H), 6.89 (d, J=6.0 Hz, 2H), 6.73 (m, 1H), 6.38 (s,
1H), 5.44 (s, 1H), 4.91 (s, 1H), 4.68 (m, 1H), 1.33 (d, J=7.1 Hz,
3H).
Example 9
##STR00023##
[0138] MS 427.2 (M+H), 449.2 (M+Na).
[0139] Anal Calcd for
C.sub.22H.sub.20F.sub.2N.sub.4O.sub.3.3/4H.sub.2O: C, 60.06; H,
4.93; N, 12.74. Found: C, 60.27; H, 4.80; N, 12.83.
[0140] .sup.1H NMR (CDCl.sub.3, 300 MHz) .delta. 12.10 (s, 1H),
7.91 (d, J=9.9 Hz, 1H), 7.32 (m, 5H), 7.17 (d, J=6.0 Hz, 2H), 7.00
(d, J=7.0 Hz, 1H), 6.72 (m, 2H), 6.62 (s, 1H), 5.67 (s(br), 1H),
5.20 (s, 1H), 4.87 (m, 1H), 1.44 (d, J=6.6 Hz, 3H).
Example 10
##STR00024##
[0142] A solution of 90480 (0.74 mmole) in 15 mL of absolute EtOH
was prepared. To the solution, was added 0.32 g of 10% Pd on
carbon. The reaction mixture was stirred under a balloon of H.sub.2
for 1 hr. The reaction mixture was filtered through a bed of celite
on a fritted filter. The filtrate was concentrated by rotary
evaporation. The crude material was flash chromatographed on silica
using 10% MeOH/CH.sub.2Cl.sub.2 as eluant to afford the reduced
product 90603 (70%).
[0143] MS 457.9 (M+H), 479.5 (M+Na).
[0144] Anal Calcd for
C.sub.23H.sub.25F.sub.2N.sub.5O.sub.3.5/4H.sub.2O; C, 57.55; H,
5.77; N, 14.59. Found: C, 57.33; H, 5.41; N, 14.26.
[0145] .sup.1H NMR (CDCl.sub.3, 300 MHz) .delta. 11.81 (s, 1H),
7.84 (d, J=9.3 Hz, 1H), 6.95 (m, 2H), 6.71 (m, 1H), 6.53 (m, 3H),
5.80 (s(br), 1H), 4.74 (m, 2H), 3.68 (s(br), 1H), 1.61 (s, 3H),
1.57 (s, 3H), 1.36 (d, J=6.6 Hz, 3H).
Example 11
##STR00025##
[0147] A solution of 0.21 mmole (1.0 eq.) of 90603 in 1 mL of
pyridine was prepared. To the solution, was added 0.53 mmole (2.5
eq.) of acetic anhydride. After stirring the solution for 3.5 days
at rt, it was poured into 25 mL of 2 M HCl. The solution was
extracted with EtOAc (3.times.40 mL). The combined organic layers
were dried over Na.sub.2SO.sub.4 and vacuum filtered. The filtrate
was rotary evaporated. The residual solid was dissolved in 5 mL of
MeOH and 0.33 mmole (2 eq.) of LiOH was added to remove undesired
acylation of the hydroxyl group. The mixture was stirred at rt for
20 min and rotary evaporated. The solid was dissolved in 15 mL of
H.sub.2O. The aqueous layer was extracted with EtOAc (3.times.40
mL). The combined organic extracts were dried over Na.sub.2SO.sub.4
and vacuum filtered. The filtrate was rotary evaporated. The crude
material was flash chromatographed on silica using 10%
MeOH/CH.sub.2Cl.sub.2 as eluant to give acylated product 91493
(50%).
[0148] MS 499.5 (M+H), 521.3 (M+Na).
[0149] Anal Calcd for
C.sub.25H.sub.27F.sub.2N.sub.5O.sub.4.3/2H.sub.2O: C, 57.03; H,
5.74; N, 13.30. Found: C, 57.03; H, 5.41; N, 12.98.
[0150] .sup.1H NMR (CD.sub.3OD, 300 MHz) .delta. 7.74 (d, J=6.6 Hz,
2H), 7.22 (d, J=6.6 Hz, 2H), 7.14 (d, J=7.1 Hz, 2H), 6.90 (m, 1H),
6.46 (s(br), 1H), 5.11 (s, 1H), 4.55 (m, 1H), 2.12 (m, 3H), 1.70
(s, 6H), 1.44 (d, J=5.5 Hz, 3H).
Example 12
##STR00026##
[0152] MS 487.3 (M+H), 509.3 (M+Na).
[0153] Anal Calcd for C.sub.25H.sub.25F.sub.3N.sub.4O.sub.3: C,
61.72; H, 5.18; N, 11.52. Found: C, 61.89; H, 5.43; N, 11.14.
[0154] .sup.1H NMR (CDCl.sub.3, 300 MHz) .delta. 12.11 (s, 1H),
11.84 (s(br), 1H), 7.87 (d, J=9.9 Hz, 1H), 7.16 (m, 4H), 6.82 (m,
3H), 6.39 (s, 1H), 5.99 (s(br), 1H), 5.15 (s, 1H), 4.74 (m, 1H),
2.31 (m, 2H), 2.09 (m, 2H), 1.71 (m, 4H), 1.43 (d, J=6.6 Hz,
3H).
Example 13
##STR00027##
[0156] MS 503.1 (M+H), 525.1 (M+Na).
[0157] Anal Calcd for
C.sub.25H.sub.25F.sub.2ClN.sub.4O.sub.3.11/10H.sub.2O: C, 57.44; H,
5.24; N, 10.72. Found: C, 57.79; H, 5.04; N, 10.32.
[0158] .sup.1H NMR (CDCl.sub.3, 300 MHz) .delta. 12.12 (s, 1H),
11.88 (s(br), 1H), 7.90 (d, J=9.9 Hz, 1H), 6.82 (m, 1H), 6.40 (s,
1H), 6.07 (s(br), 1H), 5.17 (s, 1H), 4.74 (m, 1H), 2.33 (m, 2H),
2.08 (m, 2H), 1.68 (m, 4H), 1.44 (d, J=6.6 Hz, 3H).
Example 14
##STR00028##
[0160] .sup.1H NMR (CDCl.sub.3, 300 MHz) .delta. 11.71 (s, 1H),
7.79 (d, J=9.3 Hz, 1H), 7.21 (m, 5H), 7.06 (d, J=6.6 Hz, 2H), 6.77
(m, 1H), 6.43 (s, 1H), 5.11 (s, 1H), 4.71 (m, 1H), 1.58 (d, J=6.6
Hz, 3H), 1.39 (d, J=6.6 Hz, 3H).
Example 15
##STR00029##
[0162] MS 483.3 (M+H), 505.3 (M+Na), 521.3 (M+K).
[0163] Anal Calcd for
C.sub.26H.sub.28F.sub.2N.sub.4O.sub.3.5/4H.sub.2O: C, 61.83; H,
6.09; N, 11.09. Found: C, 62.10; H, 5.97; N, 10.76.
[0164] .sup.1H NMR (CDCl.sub.3, 300 MHz) .delta. 12.06 (s, 1H),
11.75 (s(br), 1H), 7.83 (d, J=9.9 Hz, 1H), 7.24 (m, 5H), 7.20 (d,
J=6.6 Hz, 2H), 6.79 (m, 1H), 6.44 (s, 1H), 5.84 (s(br), 1H), 5.11
(s, 1H), 4.76 (m, 1H), 2.17 (m, 4H), 1.51 (m, 9H).
Example 16
##STR00030##
[0166] MS 486.5 (M+H), 508.5 (M+Na).
[0167] Anal Calcd for C.sub.25H.sub.25F.sub.3N.sub.4O.sub.3: C,
61.72; H, 5.18; N, 11.52. Found: C, 61.45; H, 5.26; N, 11.41.
[0168] .sup.1H NMR (CDCl.sub.3, 300 MHz) .delta. 11.51 (s(br), 1H),
7.60 (m, 1H), 7.34 (m, 1H), 7.06 (m, 4H), 6.90 (m, 1H), 6.79 (m,
1H), 6.47 (s, 1H), 5.12 (s, 1H), 4.73 (m, 1H), 2.52 (m, 2H), 2.17
(m, 2H), 1.80 (m, 4H), 1.39 (d, J=6.6 Hz, 3H).
Example 17
##STR00031##
[0170] MS 461.0 (M+H), 482.2 (M+Na).
[0171] Anal Calcd for C.sub.23H.sub.23F.sub.3N.sub.4O.sub.3: C,
59.99; H, 5.03; N, 12.17. Found: C, 60.08; H, 5.11; N, 11.96.
[0172] .sup.1H NMR (CD.sub.3OD, 300 MHz) .delta. 7.31 (m, 2H), 7.15
(m, 3H), 7.03 (m, 1H), 6.89 (m, 1H), 6.38 (s(br), 1H), 5.12 (s,
1H), 4.57 (m, 1H), 1.76 (s, 6H), 1.44 (d, J=7.1 Hz, 3H).
Example 18
##STR00032##
[0174] MS 471.4 (M+H), 493.2 (M+Na).
[0175] Anal Calcd for C.sub.25H.sub.28F.sub.2N.sub.4O.sub.3: C,
63.82; H, 6.00; N, 11.91. Found: C, 63.45; H, 5.92; N, 11.76.
[0176] .sup.1H NMR (CD.sub.3OD, 300 MHz) .delta. 7.15 (d, J=6.6 Hz,
2H), 6.90 (m, 4H), 6.46 (s, 1H), 5.12 (s, 1H), 4.57 (m, 1H), 2.28
(s, 6H), 1.69 (s, 6H), 1.45 (d, J=7.1 Hz, 3H).
Example 19
##STR00033##
[0178] MS 479.0 (M+H), 501.2 (M+Na).
[0179] Anal Calcd for
C.sub.23H.sub.22F.sub.4N.sub.4O.sub.3.3/2H.sub.2O: C, 54.65; H,
4.99; N, 11.08. Found: C, 54.95; H, 5.00; N, 10.77.
[0180] .sup.1H NMR (CD.sub.3OD, 300 MHz) .delta. 7.33 (m, 1H), 7.14
(d, J=6.0 Hz, 2H), 6.92 (m, 3H), 6.39 (s, 1H), 5.12 (s, 1H), 4.56
(m, 1H), 1.75 (s, 6H), 1.44 (d, J=7.1 Hz, 3H).
Example 20
##STR00034##
[0182] MS 457.2 (M+H), 478.8 (M+Na).
[0183] Anal Calcd for
C.sub.24H.sub.26F.sub.2N.sub.4O.sub.3.1/2H.sub.2O: C, 61.93; H,
5.85; N, 12.04. Found: C, 61.75; H, 5.84; N, 11.88.
[0184] .sup.1H NMR (CDCl.sub.3, 300 MHz) .delta. 11.87 (s, 1H),
7.81 (d, J=9.9 Hz, 1H), 7.18 (m, 3H), 7.08 (m, 3H), 6.83 (m, 1H),
6.11 (s, 1H), 5.07 (s, 1H), 4.53 (m, 1H), 2.81 (d, J=5.3 Hz, 1H),
2.67 (m, 1H), 1.38 (d, J=6.6 Hz, 3H), 1.27 (m, 6H).
Example 21
##STR00035##
[0186] MS 511.0 (M+H), 533.2 (M+Na).
[0187] Anal Calcd for C.sub.24H.sub.23F.sub.5N.sub.4O.sub.3: C,
56.47; H, 4.54; N, 10.98. Found: C, 56.14; H, 4.72; N, 10.79.
[0188] .sup.1H NMR (CDCl.sub.3, 300 MHz) .delta. 12.18 (s, 1H),
7.97 (d, J=9.9 Hz, 1H), 7.47 (d, J=8.2 Hz, 2H), 7.31 (d, J=8.2 Hz,
2H), 7.03 (d, J=6.5 Hz, 2H), 6.76 (m, 1H), 6.47 (s, 1H), 5.10 (s,
1H), 4.76 (m, 1H), 1.67 (d, J=3.9 Hz, 6H), 1.45 (d, J=6.6 Hz,
3H).
Example 22
##STR00036##
[0190] MS 491.4 (M+H), 513.4 (M+Na), 529.4 (M+K).
[0191] Anal Calcd for C.sub.27H.sub.24F.sub.5N.sub.4O.sub.3: C,
66.11; H, 4.93; N, 11.42. Found: C, 65.97; H, 4.91; N, 11.18.
[0192] .sup.1H NMR (CDCl.sub.3, 300 MHz) .delta. 11.89 (s(br), 1H),
7.88 (d, J=9.3 Hz, 1H), 7.44 (m, 1H), 7.28 (m, 2H), 7.10 (m, 6H),
6.96 (d, J=7.1 Hz, 2H), 6.64 (m, 2H), 5.09 (s, 1H), 4.82 (m, 1H),
4.07 (m, 2H) 1.48 (d, J=6.6 Hz, 3H).
Example 23
##STR00037##
[0194] MS 414.0 (M+H), 436.0 (M+Na)
[0195] .sup.1H NMR (CDCl.sub.3, 300 MHz) .delta. 12.08 (s, 1H),
8.74 (d, J=9.9 Hz, 1H), 8.53 (d, J=4.4 Hz, 1H), 7.91 (d, J=8.2 Hz,
1H), 7.80 (m, 1H), 7.42 (m, 1H), 6.86 (d, J=7.1 Hz, 2H), 6.73 (s,
1H), 6.60 (m, 1H), 5.10 (m, 1H). 1.81 (s, 3H), 1.76 (s, 3H), 1.56
(d, J=6.6 Hz, 3H).
Example 24
##STR00038##
[0197] MS 464.0 (M+H), 486.2 (M+Na).
[0198] Anal Calcd for
C.sub.25H.sub.23F.sub.2N.sub.5O.sub.2.3/4H.sub.2O: C, 62.94; H,
5.18; N, 14.68. Found: C, 63.09; H, 5.05; N, 14.77.
[0199] .sup.1H NMR (CDCl.sub.3, 300 MHz) .delta. 12.16 (s, 1H),
9.32 (d, J=8.8 Hz, 1H), 9.01 (d, J=9.9 Hz, 1H), 8.48 (d, J=5.5 Hz,
1H), 7.83 (m, 2H), 7.70 (t, J=7.7 Hz, 1H), 7.54 (t, J=8.2 Hz, 1H),
6.86 (d, J=6.0 Hz, 2H), 6.76 (s, 1H), 6.59 (m, 1H), 5.16 (m, 1H),
1.80 (s, 3H), 1.78 (s, 3H), 1.62 (d, J=6.6 Hz, 3H).
Example 25
##STR00039##
[0201] MS 357.2 (M+H).
[0202] Anal Calcd for C.sub.20H.sub.28N.sub.4O.sub.2: C, 67.39; H,
7.92; N, 15.72. Found: C, 67.05; H, 7.65; N, 15.72.
[0203] .sup.1H NMR (CDCl.sub.3, 300 MHz) .delta. 11.95 (s, 1H),
11.87 (s(br), 1H), 7.23 (m, 5H), 6.87 (d, J=9.3 Hz, 1H), 6.63 (s,
1H), 4.94 (m, 1H), 1.99 (m, 3H), 1.76 (s, 3H), 1.74 (s, 3H), 1.41
(d, J=6.6 Hz, 3H), 0.86 (d, J=6.0 Hz, 3H), 0.80 (d, J=5.5 Hz,
3H).
Example 26
##STR00040##
[0205] MS 343.1 (M+H).
[0206] .sup.1H NMR (CDCl.sub.3, 300 MHz) .delta. 11.86 (s, 1H),
7.23 (m, 5H), 6.60 (m, 2H), 6.63 (s, 1H), 4.93 (m, 1H), 2.19 (m,
2H), 1.75 (s, 3H), 1.74 (s, 3H), 1.50 (m, 2H), 1.40 (d, J=6.6 Hz,
3H), 0.84 (t, J=7.7 Hz, 3H).
Example 27
##STR00041##
[0208] MS 486.1 (M+H).
[0209] .sup.1H NMR (CDCl.sub.3, 300 MHz) .delta. 11.91 (s, 1H),
7.86 (d, J=8.8 Hz, 1H), 7.07 (m, 4H), 6.74 (m, 1H), 6.61 (d, J=8.8
Hz, 2H), 6.49 (s, 1H), 5.06 (s, 1H), 4.75 (m, 1H), 2.83 (s, 6H),
1.63 (s, 6H), 1.40 (d, J=6.6 Hz, 3H).
Example 28
##STR00042##
[0211] MS 478.1 (M+H).
[0212] .sup.1H NMR (CDCl.sub.3, 300 MHz) .delta. 7.22 (m, 4H), 6.57
(s, 1H), 4.86 (m, 1H), 3.78 (s(br), 2H), 2.88 (s, 3H), 1.72 (s,
3H), 1.71 (s, 3H), 1.40 (m, 12H).
Example 29
##STR00043##
[0214] MS 358.1 (M+H).
[0215] .sup.1H NMR (CDCl.sub.3, 300 MHz) .delta. 11.82 (s, 1H),
7.89 (d, J=9.9 Hz, 1H), 7.22 (m, 5H), 6.65 (s, 1H), 4.89 (m, 1H),
2.84 (s, 2H), 2.23 (s, 6H), 1.75 (s, 3H), 1.74 (s, 3H), 1.39 (d,
J=7.1 Hz, 3H).
Example 30
##STR00044##
[0217] MS 432.1 (M+H).
[0218] .sup.1H NMR (CD.sub.3OD, 300 MHz) .delta. 7.31 (m, 9H), 6.50
(s(br), 1H), 4.68 (m, 1H), 4.19 (m, 2H), 3.81 (m, 1H), 3.12 (m,
2H), 1.85 (s, 6H), 1.62 (s, 3H).
Example 31
##STR00045##
[0220] MS 378.1 (M+H).
[0221] .sup.1H NMR (CD.sub.3OD, 300 MHz) .delta. 7.38 (m, 4H), 6.46
(s, 1H), 4.69 (m, 1H), 3.65 (s, 2H), 2.64 (s, 3H), 1.80 (s, 6H),
1.57 (d, J=7.1 Hz, 3H).
Example 32
##STR00046##
[0223]
2-[2-(3,5-Difluoro-phenyl)-2-hydroxy-acetylamino]-N-[5-(1,1-dimethy-
l-2-phenyl-ethyl)-2H-pyrazol-3-yl]-propionamide MS 447 (M+H), 470
(M+Na)
[0224] MS 447 (M+H), 470 (M+Na)
[0225] .sup.1H NMR (CDCl.sub.3, 300 MHz) .delta. 7.13 (m, 5H), 6.88
(m, 3H), 6.28 (s, 1H), 5.096 (s, 1H), 4.55 (m, J=6.9, 1H), 2.87 (s,
2H), 1.41 (d, j=6.9 Hz, 3H), 1.283 (s, 6H)
Example 33
##STR00047##
[0226]
2-[2-(3,5-Difluoro-phenyl)-2-hydroxy-acetylamino]-N-[5-(1,1-dimethy-
l-2-phenyl-ethyl)-2H-pyrazol-3-yl]-propionamide
[0227] MS 447 (M+H), 470 (M+Na)
Example 34
##STR00048##
[0228]
2-[2-(3,5-Difluoro-phenyl)-2-hydroxy-acetylamino]-N-[5-(1-phenyl-et-
hyl)-2H-pyrazol-3-yl]-propionamide
[0229] MS 429 (M+H), 452 (M+Na)
Example 35
##STR00049##
[0230]
2-[2-(3,5-Difluoro-phenyl)-2-hydroxy-acetylamino]-N-{5-[1-(3,5-difl-
uoro-phenyl)-1-methyl-ethyl]-2H-pyrazol-3-yl}-propionamide
[0231] MS 479 (M+H)
[0232] .sup.1H NMR (MeOH, 300 MHz) .delta. 7.11 (m, 2H), 6.81 (m,
3H), 6.43 (s, 1H), 5.09 (s, 1H), 4.54 (m, J=7.2, 1H), 1.67 (s, 6H),
1.41 (d, j=7.2 Hz, 3H)
Example 36
##STR00050##
[0233]
2-[2-(3,5-Difluoro-phenyl)-2-hydroxy-acetylamino]-N-{5-[1-(4-fluoro-
-phenyl)-1-methyl-ethyl]-2H-pyrazol-3-yl}-propionamide
[0234] MS 461 (M+H)
[0235] .sup.1H NMR (MeOH, 300 MHz) .delta. 7.27 (m, 2.3), 7.12 (m,
2H), 7.00 (m, 2H), 6.85 (m, 1H), 6.40 (s, 1H), 5.09 (s, 1H), 4.53
(m, 1H), 1.68 (s, 6H), 1.42 (d, J=6.9 Hz, 3H)
Example 37
##STR00051##
[0236]
N-{1-[5-(1,1-Dimethyl-2-phenyl-ethyl)-2H-pyrazol-3-ylcarbamoyl]-eth-
yl}-2-hydroxy-3-methyl-butyramide
[0237] MS 387 (M+H)
[0238] .sup.1H NMR (DMSO, 300 MHz) .delta. 7.13 (m, 3H), 6.87 (m,
2H), 6.29 (s, 1H), 4.63 (m, 1H), 3.90 (d, J=3.6 Hz, 1H), 2.87 (s,
2H), 2.11 (m, 1H), 1.98 (d, J=3.9 Hz, 1H), 1.42 (d, J=6.9 Hz, 3H),
1.28 (s, 6H), 1.01 (d, J=6.9 Hz, 3H), 0.86 (d, J=6.6 Hz, 3H)
Example 38
##STR00052##
[0239]
N-(1-{5-[1-(3,5-Difluoro-phenyl)-1-methyl-ethyl]-2H-pyrazol-3-ylcar-
bamoyl}-ethyl)-2-hydroxy-3-methyl-butyramide
[0240] MS 409 (M+H)
[0241] .sup.1H NMR (DMSO, 300 MHz) .delta. 6.84 (m, 3H), 6.44 (s,
1H), 4.62 (m, 1H), 3.90 (d, J=1.5, 1H), 2.1 (m, 1H), 1.67 (s, 6H),
1.43 (d, J=6.9 Hz, 3H), 1.00 (d, J=6.9 Hz, 3H), 0.85 (d, J=6.6 Hz,
3H)
Example 39
##STR00053##
[0242]
2-[2-(3,5-Difluoro-phenyl)-2-hydroxy-acetylamino]-N-[5-(4-phenyl-te-
trahydro-pyran-4-yl)-2H-pyrazol-3-yl]-propionamide
[0243] MS 485 (M+H), 506.8 (M+Na)
[0244] .sup.1H NMR (MeOH, 300 MHz) .delta. 7.25 (m, 4H), 7.09 (m,
3H), 6.85 (m, 1H), 6.44 (s, 1H), 5.06 (s, 1H), 4.51 (m, 1H), 3.78
(m, 2H), 3.60 (m, 2H), 2.27 (m, 4H), 1,36 (d, J=6.9 Hz, 3H)
Example 40
##STR00054##
[0245]
2-[2-(3,5-Difluoro-phenyl)-2-hydroxy-acetylamino]-N-{5-[1-(3,5-difl-
uoro-phenyl)-1-methyl-ethyl]-2H-pyrazol-3-yl}-propionamide
[0246] MS 479 (M+H)
Example 41
##STR00055##
[0247]
2-Chloro-N-(1-{5-[1-(3,5-difluoro-phenyl)-1-methyl-ethyl]-2H-pyrazo-
l-3-ylcarbamoyl}-ethyl)-benzamide
[0248] MS 446.8 (M+H), 468.8 (M+Na), 485 (M+K)
[0249] Anal Calcd for C.sub.22H.sub.21ClF.sub.2N.sub.4O.sub.2: C,
59.13; H, 4.74; N, 12.54 Found: C, 58.88; H, 4.94; N, 12.19
[0250] .sup.1H NMR (MeOH, 300 MHz) .delta. 7.50 (m, 1H), 7.33 (m,
3H), 6.71 (m, 2H), 6.58 (m, 1H), 4.77 (m, 1H), 1.62 (s, 6H), 1.47
(d, J=9 Hz, 3H)
Example 42
##STR00056##
[0251]
N-(1-{5-[1-(3,5-Difluoro-phenyl)-1-methyl-ethyl]-2H-pyrazol-3-ylcar-
bamoyl}-ethyl)-2-trifluoromethyl-benzamide
[0252] MS 481 (M+H), 503 (M+Na), 518 (M+K)
[0253] Anal Calcd for C.sub.23H.sub.21F.sub.5N.sub.4O.sub.2: C,
57.50; H, 4.41; N, 11.66 Found: C, 57.20; H, 4.57; N, 11.23
[0254] .sup.1H NMR (MeOH, 300 MHz) .delta. 7.66 (m, 4H), 6.79 (m,
3H), 6.51 (m, 1H), 4.67 (m, 1H), 1.66 (s, 6H), 1.46 (d, J=8.7 Hz,
3H)
Example 43
##STR00057##
[0255]
N-(1-{5-[1-(3,5-Difluoro-phenyl)-1-methyl-ethyl]-2H-pyrazol-3-ylcar-
bamoyl}-ethyl)-3-trifluoromethyl-benzamide
[0256] MS 481 (M+H), 503 (M+Na), 518.8 (M+K)
[0257] Anal Calcd for C.sub.23H.sub.21F.sub.5N.sub.4O.sub.2: C,
57.50; H, 4.41; N, 11.66 Found: C, 57.13, H, 4.46; N, 11.41
[0258] .sup.1H NMR (MeOH, 300 MHz) .delta. 8.19 (s, 1H), 8.10 (d,
J=7.8 Hz, 1H), 7.80 (d, J=7.8 Hz, 1H) 7.61 (t, J=7.8 Hz, 1H), 6.752
(m, 3H), 6.43 (s, 1H), 4.72 (m, 1H), 1.64 (s, 6H), 1.52 (d, J=7.2
Hz, 3H)
Example 44
##STR00058##
[0259]
N-{5-[1-(3,5-Difluoro-phenyl)-1-methyl-ethyl]-2H-pyrazol-3-yl}-2-[2-
-hydroxy-2-(2-trifluoromethyl-phenyl)-acetylamino]propionamide
[0260] MS 511.2 (M+H), 533 (M+Na)
[0261] .sup.1H NMR (MeOH, 300 MHz) .delta. 7.59 (m, 3H), 7.45 (m,
1H), 6.79 (m, 3H), 6.39 (m, 1H), 5.43 (m, 1H), 5.57 (m, 1H), 1.66
(s, 7H), 1.48 (d, J=6.9 Hz, 3H)
Example 45
##STR00059##
[0262]
2-[2-(4-tert-Butyl-phenyl)-2-hydroxy-acetylamino]-N-{5-[1-(3,5-difl-
uoro-phenyl)-1-methyl-ethyl]-2H-pyrazol-3-yl}-propionamide
[0263] MS 499.2 (M+H), 521 (M+Na)
[0264] .sup.1H NMR (MeOH, 300 MHz) .delta. 7.36 (s, 4H), 6.82 (m,
3H), 6.41 (s, 1H), 5.01 (s, 1H), 4.53 (m, 1H), 1.66 (s, 6H), 1.41
(d, J=6.9 Hz, 3H), 1.28 (s, 9H)
Example 46
##STR00060##
[0265]
2-[2-(4-tert-Butyl-phenyl)-2-hydroxy-acetylamino]-N-{5-[1-(3,5-difl-
uoro-phenyl)-1-methyl-ethyl]-2H-pyrazol-3-yl}-propionamide
[0266] MS 499.2 (M+H), 521 (M+Na)
[0267] .sup.1H NMR (MeOH, 300 MHz) .delta. 7.35 (s, 4H), 6.79 (m,
3H), 6.38 (s, 1H), 5.01 (s, 1H), 5.46 (m, 1H), 1.65 (s, 6H), 1.43
(d, J=6.9 Hz, 3H), 1.26 (s, 9H)
Example 47
##STR00061##
[0268]
2-[2-(3-Chloro-phenyl)-2-hydroxy-acetylamino]-N-{5-[1-(3,5-difluoro-
-phenyl)-1-methyl-ethyl]-2H-pyrazol-3-yl}-propionamide
[0269] MS 477.2 (M+H), 499.0 (M+Na), 514.8 (M+K)
[0270] Anal Calcd for C.sub.23H.sub.23ClF.sub.2N.sub.4O.sub.3: C,
57.92; H, 4.86; N, 11.75 Found: C, 57.75; H, 5.27; N, 11.23
[0271] .sup.1H NMR (MeOH, 300 MHz) .delta. 7.47 (s, 1H), 7.37 (m,
1H), 7.27 (m, 2H), 6.78 (m, 3H), 6.42 (s, 1H), 5.04 (s, 1H), 4.51
(m, 1H), 1.65 (s, 6H), 1.40 (d, J=7.2 Hz, 3H)
Example 48
##STR00062##
[0272]
N-{5-[1-(3,5-Difluoro-phenyl)-1-methyl-ethyl]-2H-pyrazol-3-yl}-2-(2-
-hydroxy-2-p-tolyl-acetylamino)-propionamide
[0273] MS 457.2 (M+H), 479 (M+Na)
[0274] .sup.1H NMR (MeOH, 300 MHz) .delta. 7.30 (d, J=8.1 Hz, 2H),
7.13 (d, J=7.8 Hz, 2H), 6.79 (m, 3H), 6.43 (s, 1H), 4.99 (s, 1H),
4.53 (m, 1H), 2.28 (s, 3H), 1.654 (s, 6H), 1.40 (d, J=6.9 Hz,
3H)
Example 49
##STR00063##
[0275]
2-[2-(4-Chloro-phenyl)-2-hydroxy-acetylamino]-N-{5-[1-(3,5-difluoro-
-phenyl)-1-methyl-ethyl]-2H-pyrazol-3-yl}-propionamide
[0276] MS 477.0 (M+H), 498.8 (M+Na), 514.8 (M+K)
[0277] Anal Calcd for C.sub.23H.sub.23ClF.sub.2N.sub.4O.sub.3: C,
57.92; H, 4.86; N, 11.75 Found: C, 57.70; H, 4.92; N, 11.43
[0278] .sup.1H NMR (MeOH, 300 MHz) .delta. 7.43 (d, J=8.4 Hz, 2H),
7.30 (d, J=10.8 Hz, 2H), 6.78 (m, 3H), 6.40 (s, 1H), 5.04 (s, 1H),
4.52 (m, 1H), 1.647 (s, 6H), 1.39 (d, J=7.2 Hz, 3H)
Example 50
##STR00064##
[0279]
N-{5-[1-(3,5-Difluoro-phenyl)-1-methyl-ethyl]-2H-pyrazol-3-yl}-2-[2-
-hydroxy-2-(4-methoxy-phenyl)-acetylamino]-propionamide
[0280] MS 473 (M+H), 495 (M+Na)
[0281] Anal Calcd for C.sub.24H.sub.26F.sub.2N.sub.4O.sub.4: C,
61.01; H, 5.55; N, 11.86 Found: C, 61.63; H, 5.25; N, 11.11
[0282] .sup.1H NMR (MeOH, 300 MHz) .delta. 7.32 (m, 2H), 6.83 (m,
5H), 6.45 (s, 1H), 4.98 (s, 1H), 4.51 (m, 1H), 3.75 (s, 3H), 1.66
(s, 6H), 1.41 (d, J=6.9 Hz, 3H)
Example 51
##STR00065##
[0283]
N-{5-[1-(3,5-Difluoro-phenyl)-1-methyl-ethyl]-2H-pyrazol-3-yl}-2-[2-
-hydroxy-2-(4-trifluoromethyl-phenyl)-acetylamino]-propionamide
[0284] MS 511 (M+H), 532.8 (M+Na)
[0285] Anal Calcd for C.sub.24H.sub.23F.sub.5N.sub.4O.sub.3: C,
56.47; H, 4.54; N, 10.98 Found: C, 56.12; H, 4.68; N, 10.53
[0286] .sup.1H NMR (MeOH, 300 MHz) .delta. 7.64 (m, 4H), 6.78 (m,
3H), 6.48 (s, 1H), 5.14 (s, 1H), 4.52 (m, 1H), 1.65 (s, 6H), 1.39
(d, J=6.9 Hz, 3H)
Example 52
##STR00066##
[0287]
2-[2-(2-Chloro-phenyl)-2-hydroxy-acetylamino]-N-{5-[1-(3,5-difluoro-
-phenyl)-1-methyl-ethyl]-2H-pyrazol-3-yl}-propionamide
[0288] MS 477 (M+H)
[0289] .sup.1H NMR (MeOH, 300 MHz) .delta. 7.36 (m, 2H), 7.27 (m,
2H), 6.79 (m, 3H), 6.51 (s, 1H), 5.51 (s, 1H), 4.57 (m, 1H), 1.66
(s, 6H), 1.46 (d, J=6.9 Hz, 3H)
Example 53
##STR00067##
[0290]
N-{5-[1-(4-Chloro-phenyl)-1-methyl-ethyl]-2H-pyrazol-3-yl}-2-(2-hyd-
roxy-cyclohexylamino)-propionamide
[0291] MS 406 (M+H)
[0292] .sup.1H NMR (CDCl.sub.3, 300 MHz) .delta. 7.24 (m, 4H), 6.53
(s, 1H), 4.67 (m, 6H), 3.82 (m, 1H), 2.66 (m, 1H), 2.09 (m, 1H),
1.58 (m, 13H), 1.17 (m, 3H)
Example 54
##STR00068##
[0293]
N-(1-{5-[1-(4-Fluoro-phenyl)-1-methyl-ethyl]-2H-pyrazol-3-ylcarbamo-
yl}-ethyl)-2-hydroxy-3-methyl-butyramide
[0294] MS 391 (M+H)
[0295] .sup.1H NMR (MeOH, 300 MHz) .delta. 7.27 (m, 2H), 6.99 (m,
2H), 6.40 (s, 1H), 4.59 (m, 1H), 3.89 (d, J=3.6 Hz, 1H), 2.11 (m,
1H), 1.69 (s, 6H), 1.44 (d, J=7.2 Hz, 3H), 1.02 (d, J=6.9 Hz, 3H),
0.87 (d, J=6.9 Hz, 3H)
Example 55
##STR00069##
[0296]
N-{2-tert-Butyl-5-[1-(4-fluoro-phenyl)-1-methyl-ethyl]-2H-pyrazol-3-
-yl}-2-[2-(3,5-difluoro-phenyl)-2-hydroxy-acetylamino]-propionamide
[0297] MS 517 (M+H)
[0298] .sup.1H NMR (CDCl.sub.3, 300 MHz) .delta. 8.18 (s, 1H), 7.24
(m, 4H), 6.96 (m, 2H), 6.91 (m, 2H), 6.77 (m, 1H), 5.97 (s, 1H),
5.01 (s, 1H), 4.49 (m, 1H), 1.61 (s, 6H), 1.56 (s, 9H), 1.38 (d,
J=7.2 Hz, 3H)
Example 56
##STR00070##
[0299]
N-{2-tert-Butyl-5-[1-(3,5-difluoro-phenyl)-1-methyl-ethyl]-2H-pyraz-
ol-3-yl}-2-[2-(3,5-difluoro-phenyl)-2-hydroxy-acetylamino]-propionamide
[0300] MS 535 (M+H)
[0301] .sup.1H NMR (CDCl.sub.3, 300 MHz) .delta. 8.44 (s, 1H), 7.38
(d, J=7.8 Hz, 1H), 6.93 (m, 2H), 6.75 (m, 3H), 6.54 (m, 1H), 5.96
(s, 1H), 4.97 (s, 1H), 4.64 (s, 1H), 4.52 (m, 1H), 1.58 (s, 6H),
1.51 (s, 9H), 1.23 (d, J=6.9 Hz, 3H)
Example 57
##STR00071##
[0302]
N-[2-tert-Butyl-5-(1,1-dimethyl-2-phenyl-ethyl)-2H-pyrazol-3-yl]-2--
[2-(3,5-difluoro-phenyl)-2-hydroxy-acetylamino]-propionamide
[0303] MS 513 (M+H)
[0304] .sup.1H NMR (CDCl.sub.3, 300 MHz) .delta. 8.30 (s, 1H), 7.41
(d, J=7.5 Hz, 1H), 7.14 (m, 3H), 6.97 (m, 4H), 6.72 (m, 1H), 6.01
(s, 1H), 5.00 (s, 1H), 4.52 (m, 1H), 2.85 (s, 2H), 1.52 (s, 9H),
1.38 (d, J=6.9 Hz, 3H), 1.18 (s, 6H)
Example 58
##STR00072##
[0305]
N-[2-tert-Butyl-5-(1,1-dimethyl-2-phenyl-ethyl)-2H-pyrazol-3-yl]-2--
[2-(3,5-difluoro-phenyl)-2-hydroxy-acetylamino]-propionamide
[0306] MS 513 (M+H)
[0307] .sup.1H NMR (CDCl.sub.3, 300 MHz) .delta. 8.15 (s, 1H), 7.43
(d, J=7.5 Hz, 1H), 7.13 (m, 3H), 6.96 (m, 4H), 6.70 (m, 1H), 5.96
(s, 1H), 5.04 (s, 1H), 4.61 (m, 1H), 4.32 (m, 1H), 2.81 (s, 2H),
1.44 (d, J=6.9 Hz, 3H), 1.17 (s, 6H)
Example 58a
##STR00073##
[0308]
{1-[2-tert-Butyl-5-(1,1-dimethyl-2-phenyl-ethyl)-2H-pyrazol-3-ylcar-
bamoyl]-ethyl}-carbamic acid tert-butyl ester
[0309] MS 443 (M+H)
[0310] .sup.1H NMR (CDCl.sub.3, 300 MHz) .delta. 7.18 (m, 3H), 6.97
(m, 2H), 6.17 (s, 1H), 4.32 (m, 1H), 2.88 (s, 2H), 1.58 (s, 9H),
1.46 (m, 12H), 1.22 (s, 6H)
Example 59
##STR00074##
[0311]
N-[2-tert-Butyl-5-(1,1-dimethyl-2-phenyl-ethyl)-2H-pyrazol-3-yl]-2--
phenylacetylamino-propionamide
[0312] MS 390 (M+H)
[0313] .sup.1H NMR (CDCl.sub.3, 300 MHz) .delta. 7.40 (m, 5H), 7.16
(m, 3H), 6.94 (m, 3H), 6.25 (s, 1H), 3.77 (s, 2H), 2.86 (s, 2H),
1.29 (s, 9H), 1.21 (s, 6H)
Example 60
##STR00075##
[0314]
N-{1-[2-tert-Butyl-5-(1,1-dimethyl-2-phenyl-ethyl)-2H-pyrazol-3-ylc-
arbamoyl]-ethyl}-butyramide
[0315] MS 342 (M+H)
[0316] .sup.1H NMR (MeOH, 300 MHz) .delta. 7.11 (m, 3H), 6.92 (m,
2H), 5.82 (s, 1H), 2.86 (s, 2H), 2.34 (t, J=6.6 Hz, 2H), 1.71 (m,
2H), 1.57 (s, 9H), 1.22 (s, 6H) 1.02 (t, J=7.5 Hz, 3H)
Example 61
##STR00076##
[0317]
N-[2-tert-Butyl-5-(4-phenyl-tetrahydro-pyran-4-yl)-2H-pyrazol-3-yl]-
-2-[2-(3,5-difluoro-phenyl)-2-hydroxy-acetylamino]-propionamide
[0318] MS 541.4 (M+H), 563.4 (M+Na)
[0319] Anal Calcd for C.sub.29H.sub.34N.sub.4F.sub.2O.sub.3: C,
64.43; H, 6.34; N, 10.36 Found: C, 64.20; H, 6.22; N, 10.30
[0320] .sup.1H NMR (MeOH, 300 MHz) .delta. 8.43 (s, 1H), 7.24 (m,
5H), 7.09 (m, 1H), 6.91 (m, 2H), 6.70 (m, 1H), 5.91 (s, 1H), 4.90
(s, 1H), 4.78 (s, 1H), 4.42 (m, 1H), 3.73 (m, 4H), 2.38 (m, 2H),
2.12 (m, 3H), 1.51 (s, 9H), 1.20 (d, J=6 Hz, 3H)
Example 62
##STR00077##
[0321]
N-(1-{2-tert-Butyl-5-[1-(3,5-difluoro-phenyl)-1-methyl-ethyl]-2H-py-
razol-3-ylcarbamoyl}-ethyl)-3-trifluoromethyl-benzamide
[0322] MS 537.2 (M+H), 559.2 (M+Na)
[0323] Anal Calcd for C.sub.27H.sub.29F.sub.5N.sub.4O.sub.2: C,
60.44; H, 5.45; N, 10.44 Found: C, 60.27; H, 5.26; N, 10.17
[0324] .sup.1H NMR (CDCl.sub.3, 300 MHz) .delta. 8.92 (s, 1H), 8.01
(s, 1H), 7.88 (d, J=7.8 Hz, 1H), 7.89 (d, J=7.5 Hz, 1H), 7.68 (d,
J=7.8 Hz, 1H), 7.39 (t, J=7.8 Hz, 1H), 6.77 (m, 2H), 6.50 (m, 1H),
5.94 (s, 1H), 5.04 (m, 1H), 1.55 (s, 6H), 1.52 (m, 12H)
Example 63
##STR00078##
[0325]
N-{2-tert-Butyl-5-[1-(3,5-difluoro-phenyl)-1-methyl-ethyl]-2H-pyraz-
ol-3-yl}-2-[2-(4-tert-butyl-phenyl)-2-hydroxy-acetylamino]-propionamide
[0326] MS 555 (M+H), 577.2 (M+Na)
[0327] Anal Calcd for C.sub.31H.sub.40F.sub.2N.sub.4O.sub.3: C,
67.13; H, 7.27; N, 10.10 Found: C, 67.07; H, 7.28; N, 9.90
[0328] .sup.1H NMR (CDCl.sub.3, 300 MHz) .delta. 8.61 (s, 1H), 7.32
(m, 3H), 7.22 (m, 2H), 6.82 (m, 2H), 6.54 (m, 1H), 5.98 (s, 1H),
4.95 (d, J=3 Hz, 1H), 4.56 (m, 1H), 3.98 (d, J=6 Hz, 1H), 1.58 (s,
6H), 1.48 (s, 9H), 1.28 (m, 12H)
Example 64
##STR00079##
[0329]
N-{2-tert-Butyl-5-[1-(3,5-difluoro-phenyl)-1-methyl-ethyl]-2H-pyraz-
ol-3-yl}-2-[2-(3-chloro-phenyl)-2-hydroxy-acetylamino]-propionamide
[0330] MS 533.01 (M+H), 555 (M+Na)
[0331] Anal Calcd for C.sub.27H.sub.31ClF.sub.2N.sub.4O.sub.3: C,
60.84; H, 5.86; N, 10.51 Found: C, 60.62; H, 6.03; N, 10.19
[0332] .sup.1H NMR (CDCl.sub.3, 300 MHz) .quadrature. 8.46 (s, 1H),
7.35 (m, 2H), 7.20 (m, 3H), 6.80 (m, 2H), 6.53 (m, 1H), 5.95 (s,
1H), 4.95 (s, 1H), 4.52 (m, 1H), 1.56 (s, 6H), 1.48 (s, 9H), 1.29
(d, J=6 Hz, 3H)
Example 65
##STR00080##
[0333]
2-(2-Cyclohexyl-2-hydroxy-acetylamino)-N-{5-[1-(3,5-difluoro-phenyl-
)-1-methyl-ethyl]-2H-pyrazol-3-yl}-propionamide
[0334] MS 449 (M+H), 471 (M+Na)
[0335] Anal. Calcd for C.sub.23H.sub.30F.sub.2N.sub.4O.sub.3: C,
61.59; H, 6.74; N, 12.49. Found: C, 61.66; H, 6.80; N, 12.28.
Example 66
##STR00081##
[0336]
N-{5-[1-(3,5-Difluoro-phenyl)-1-methyl-ethyl]-2H-pyrazol-3-yl}-2-[2-
-(4-fluoro-phenyl)-2-hydroxy-acetylamino]-propionamide
[0337] MS 461 (M+H), 483 (M+Na)
[0338] Anal. Calcd for C.sub.23H.sub.23F.sub.3N.sub.4O.sub.3: C,
60.00; H, 5.03; N, 12.17. Found: C, 60.20; H, 5.38; N, 11.96.
Example 67
##STR00082##
[0339]
N-{5-[1-(3,5-Difluoro-phenyl)-1-methyl-ethyl]-2H-pyrazol-3-yl}-2-(2-
-hydroxy-2-phenyl-acetylamino)-propionamide
[0340] MS 443 (M+H), 465 (M+Na)
[0341] Anal. Calcd for C.sub.23H.sub.24F.sub.2N.sub.4O.sub.3: C,
62.43; H, 5.47; N, 12.66. Found: C, 61.89; H, 5.67; N, 12.38.
Example 68
##STR00083##
[0342]
2-(2-Hydroxy-2-phenyl-acetylamino)-N-[5-(1-methyl-1-phenyl-ethyl)-2-
H-pyrazol-3-yl]-propionamide
[0343] MS 407 (M+H), 429 (M+Na)
[0344] Anal. Calcd for C.sub.23H.sub.26N.sub.4O.sub.3: C, 67.96; H,
6.44; N, 13.78. Found: C, 66.58; H, 6.70; N, 13.66.
Example 69
##STR00084##
[0345]
2-(2-Cyclohexyl-2-hydroxy-acetylamino)-N-[5-(1-methyl-1-phenyl-ethy-
l)-2H-pyrazol-3-yl]-propionamide
[0346] MS 413 (M+H), 435 (M+Na)
Example 70
##STR00085##
[0347]
2-[(3,5-Difluoro-phenyl)-acetylamino]-N-{5-[1-(3,5-difluoro-phenyl)-
-1-methyl-ethyl]-2H-pyrazol-3-yl}-propionamide
[0348] MS 463 (M+H), 485 (M+Na)
[0349] Anal. Calcd for C.sub.23H.sub.22F.sub.4N.sub.4O.sub.2: C,
59.74; H, 4.80; N, 12.12. Found: C, 59.51; H, 4.88; N, 11.91.
Example 71
##STR00086##
[0350]
2-(2-Cyclohexyl-acetylamino)-N-[5-(1-methyl-1-phenyl-ethyl)-2H-pyra-
zol-3-yl]-propionamide
[0351] MS 397 (M+H), 419 (M+Na)
[0352] Anal. Calcd for C.sub.23H.sub.33N.sub.4O.sub.2: C, 69.67; H,
8.13; N, 14.13. Found: C, 69.40; H, 8.20; N, 14.01.
Example 72
##STR00087##
[0353]
2-(3,5-Difluoro-phenyl)-N-(1-{5-[1-(3,5-difluoro-phenyl)-1-methyl-e-
thyl]-2H-pyrazol-3-ylcarbamoyl}-ethyl)-isobutyramide
[0354] MS 491 (M+H), 513 (M+Na)
[0355] Anal. Calcd for C.sub.25H.sub.26F.sub.4N.sub.4O.sub.2: C,
61.22; H, 5.34; N, 11.42. Found: C, 61.16; H, 5.60; N, 11.19.
Example 73
##STR00088##
[0356]
N-(1-{5-[1-(3,5-Difluoro-phenyl)-1-methyl-ethyl]-2H-pyrazol-3-ylcar-
bamoyl}-ethyl)-3-methyl-3-phenyl-butyramide
[0357] MS 469 (M+H), 491 (M+Na)
[0358] Anal. Calcd for C.sub.26H.sub.30F.sub.2N.sub.4O.sub.2: C,
66.65; H, 6.45; N, 11.96. Found: C, 66.31; H, 6.50; N, 11.89.
Example 74
##STR00089##
[0359]
(2-Benzo[1,3]dioxol-5-yl-acetylamino)-N-{5-[1-(3,5-difluoro-phenyl)-
-1-methyl-ethyl]-2H-pyrazol-3-yl}-propionamide
[0360] MS 471 (M+H), 493 (M+Na)
[0361] Anal. Calcd for C.sub.24H.sub.24F.sub.2N.sub.4O.sub.4: C,
61.27; H, 5.14; N, 11.91. Found: C, 60.72; H, 5.48; N, 11.59.
Example 75
##STR00090##
[0362]
2-[2-(4-Chloro-phenyl)-2-hydroxy-acetylamino]-N-[5-(1-methyl-1-phen-
yl-ethyl)-2H-pyrazol-3-yl]-propionamide
[0363] MS 463 (M+Na)
[0364] Anal. Calcd for C.sub.23H.sub.25ClN.sub.4O.sub.3: C, 62.65;
H, 5.71; N, 12.71. Found: C, 62.38; H, 5.97; N, 12.38.
Example 76
##STR00091##
[0365]
2-[2-(2,3-Difluoro-phenyl)-2-hydroxy-acetylamino]-N-{5-[1-(3,5-difl-
uoro-phenyl)-1-methyl-ethyl]-2H-pyrazol-3-yl}-propionamide
[0366] MS 479 (M+H)
[0367] Anal. Calcd for C.sub.23H.sub.22F.sub.4N.sub.4O.sub.3: C,
57.74; H, 4.63; N, 11.71. Found: C, 57.41; H, 4.89; N, 11.31.
Example 77
##STR00092##
[0368]
2-[2-(3,5-Dimethoxy-phenyl)-acetylamino]-N-[5-(1-methyl-1-phenyl-et-
hyl)-2H-pyrazol-3-yl]-propionamide
[0369] MS 451 (M+H), 473 (M+Na)
[0370] Anal. Calcd for C.sub.25H.sub.30N.sub.4O.sub.4: C, 66.65; H,
6.71; N, 12.44. Found: C, 66.11; H, 6.62; N, 12.29.
Example 78
##STR00093##
[0371]
2-[2-(3,5-Bis-trifluoromethyl-phenyl)-acetylamino]-N-[5-(1-methyl-1-
-phenyl-ethyl)-2H-pyrazol-3-yl]-propionamide
[0372] MS 527 (M+H)
[0373] Anal. Calcd for C.sub.25H.sub.24F.sub.6N.sub.4O.sub.2: C,
57.03; H, 4.59; N, 10.64. Found: C, 56.82; H, 4.38; N, 10.54.
Example 79
##STR00094##
[0374]
2-(2-Benzo[1,3]dioxol-5-yl-acetylamino)-N-{2-tert-butyl-5-[1-(3,5-d-
ifluoro-phenyl)-1-methyl-ethyl]-2H-pyrazol-3-yl}-propionamide
[0375] MS 527 (M+H), 549 (M+Na)
[0376] Anal. Calcd for C.sub.28H.sub.32F.sub.2N.sub.4O.sub.4: C,
63.87; H, 6.13; N, 10.64. Found: C, 63.77; H, 6.02; N, 10.53.
Example 80
##STR00095##
[0377]
N-[2-tert-Butyl-5-(1-methyl-1-phenyl-ethyl)-2H-pyrazol-3-yl]-2-[2-h-
ydroxy-2-(4-trifluoromethyl-phenyl)-acetylamino]-propionamide
[0378] MS 531 (M+H), 553 (M+Na)
[0379] Anal. Calcd for C.sub.28H.sub.33F.sub.3N.sub.4O.sub.3: C,
63.38; H, 6.27; N, 10.56. Found: C, 62.99; H, 6.56; N, 10.23.
Example 81
##STR00096##
[0380]
N-{2-tert-Butyl-5-[1-(3,5-difluoro-phenyl)-1-methyl-ethyl]-2H-pyraz-
ol-3-yl}-2-[2-(3,5-difluoro-phenyl)-acetylamino]-propionamide
[0381] MS 519 (M+H), 541 (M+Na)
[0382] Anal. Calcd for C.sub.27H.sub.30F.sub.4N.sub.4O.sub.2: C,
62.54; H, 5.83; N, 10.80. Found: C, 62.26; H, 5.74; N, 10.44.
Example 82
##STR00097##
[0383]
N-[2-tert-Butyl-5-(1-methyl-1-phenyl-ethyl)-2H-pyrazol-3-yl]-2-[2-(-
4-chloro-phenyl)-2-hydroxy-acetylamino]-propionamide
[0384] MS 497 (M+H), 519 (M+Na)
Example 83
##STR00098##
[0385]
N-(1-{2-tert-Butyl-5-[1-(3,5-difluoro-phenyl)-1-methyl-ethyl]-2H-py-
razol-3-ylcarbamoyl}-ethyl)-3-methyl-3-phenyl-butyramide
[0386] MS 525 (M+H), 547 (M+Na)
[0387] Anal. Calcd for C.sub.30H.sub.38F.sub.2N.sub.4O.sub.2: C,
68.68; H, 7.30; N, 10.68. Found: C, 68.37; H, 7.54; N, 10.52.
Example 84
##STR00099##
[0388]
N-[2-tert-Butyl-5-(1-methyl-1-phenyl-ethyl)-2H-pyrazol-3-yl]-2-(2-c-
yclohexyl-acetylamino)-propionamide
[0389] MS 453 (M+H), 475 (M+Na)
[0390] Anal. Calcd for C.sub.27H.sub.40N.sub.4O.sub.2: C, 71.65; H,
8.91; N, 12.38. Found: C, 71.35; H, 8.66; N, 11.81.
Example 85
##STR00100##
[0391]
N-{2-tert-Butyl-5-[1-(3,5-difluoro-phenyl)-1-methyl-ethyl]-2H-pyraz-
ol-3-yl}-2-[2-(2,3-difluoro-phenyl)-2-hydroxy-acetylamino]-propionamide
[0392] MS 535 (M+H), 557 (M+Na)
[0393] Anal. Calcd for C.sub.27H.sub.30F.sub.4N.sub.4O.sub.3: C,
60.67; H, 5.66; N, 10.48. Found: C, 60.27; H, 5.71; N, 10.26.
Example 86
##STR00101##
[0394]
N-{2-tert-Butyl-5-[1-(3,5-difluoro-phenyl)-1-methyl-ethyl]-2H-pyraz-
ol-3-yl}-2-(2-hydroxy-2-phenyl-acetylamino)-propionamide
[0395] MS 547 (M+H), 569 (M+Na)
[0396] Anal. Calcd for C.sub.29H.sub.34F.sub.4N.sub.4O.sub.2: C,
63.72; H, 6.27; N, 10.25. Found: C, 63.53; H, 6.05; N, 10.40.
Example 87
##STR00102##
[0397]
N-[2-tert-Butyl-5-(1-methyl-1-phenyl-ethyl)-2H-pyrazol-3-yl]-2-(2-h-
ydroxy-2-phenyl-acetylamino)-propionamide
[0398] MS 521 (M+Na)
[0399] Anal. Calcd for C.sub.27H.sub.32F.sub.2N.sub.4O.sub.3: C,
65.05; H, 6.47; N, 11.24. Found: C, 64.80; H, 6.45; N, 10.88.
Example 88
##STR00103##
[0400]
N-[2-tert-Butyl-5-(1-methyl-1-phenyl-ethyl)-2H-pyrazol-3-yl]-2-(2-c-
yclohexyl-2-hydroxy-acetylamino)-propionamide
[0401] MS 469 (M+H), 491 (M+Na)
[0402] Anal. Calcd for C.sub.27H.sub.40N.sub.4O.sub.3: C, 69.20; H,
8.60; N, 11.96. Found: C, 68.77; H, 8.22; N, 12.04.
Example 88a
##STR00104##
[0403]
N-(5-Cyclopentyl-2H-pyrazol-3-yl)-2-[2-(3,5-difluror-phenyl)2-2hydr-
oxy-acetylamino]-propionamide
[0404] MS 393 (M+H), MS 415 (M+Na).
Example 89
##STR00105##
[0405]
2-[2-(3,5-Difluorophenyl)-2-hydroxy-acetylamino]-N-[5-(1-methyl-1-p-
henyl-ethyl)-2H-pyrazol-3-yl]-propionamide
[0406] MS 443 (M+H), MS 465 (M+Na).
Example 90
##STR00106##
[0407]
(S)-2-(3,5-difluorophenyl)-2-hydroxy-N--((R)-2-oxo-2-(3-(2-phenylpr-
opan-2-yl)-1H-pyrazol-5-ylamino)-1-(thiophen-2-yl)ethyl)acetamide
[0408] MS 511 (M+H), 533 (M+Na).
Example 91
##STR00107##
[0409]
(S)-2-((S)-2-(3,5-difluorophenyl)-2-hydroxyacetylamino)-N-(3-(2-(py-
ridin-3-yl)propan-2-yl)-1H-pyrazol-5-yl)propanamide
[0410] MS 444 (M+H).
Example 92
##STR00108##
[0411]
N-{5-[1-(3-Chloro-phenyl)-1-methyl-ethyl]-2H-pyrazol-3-yl}-2-[2-(3,-
5-difluoro-phenyl)-2-hydroxy-acetylamino]-propionamide
[0412] MS 477.1 (M+H)
Example 93
##STR00109##
[0413]
2-[2-(3,5-Difluoro-phenyl)-2-hydroxy-acetylamino]-N-{5-[1-(3,4-difl-
uoro-phenyl)-1-methyl-ethyl]-2H-pyrazol-3-yl}-propionamide
[0414] MS 479.1 (M+H)
Example 94
##STR00110##
[0415]
2-[2-(3,5-Difluoro-phenyl)-2-hydroxy-acetylamino]-N-{5-[1-(3-fluoro-
-phenyl)-1-methyl-ethyl]-2H-pyrazol-3-yl}-propionamide
[0416] MS 461.1 (M+H)
Example 95
##STR00111##
[0417]
2-[2-(3,5-Difluoro-phenyl)-2-hydroxy-acetylamino]-N-{5-[1-methyl-1--
(3-trifluoromethyl-phenyl)-ethyl]-2H-pyrazol-3-yl}-propionamide
[0418] MS 511.1 (M+H)
Example 96
##STR00112##
[0419]
N-{5-[1-(3,4-Dichloro-phenyl)-1-methyl-ethyl]-2H-pyrazol-3-yl}-2-[2-
-(3,5-difluoro-phenyl)-2-hydroxy-acetylamino]-propionamide
[0420] MS 511.0 (M+H)
Example 97
##STR00113##
[0421]
2-[2-(3,5-Difluoro-phenyl)-2-hydroxy-acetylamino]-N-{5-[1-(3-methox-
y-phenyl)-1-methyl-ethyl]-2H-pyrazol-3-yl}-propionamide
[0422] MS 473.2 (M+H)
Example 98
##STR00114##
[0423]
(2S)--N-(5-Cyclopropyl-2H-pyrazol-3-yl)-2-[2-(3,5-difluorophenyl)-a-
cetylamino]-propionamide
[0424] MS 349 (M+H), 371 (M+Na).
Example 99
##STR00115##
[0425]
(2S,2'S)--N-(5-Cyclopropyl-2H-pyrazol-3-yl)-2-[2-(3,5-difluoropheny-
l)-2-hydroxy-acetylamino]-propionamide
[0426] MS
[0427] .sup.1H NMR (CDCl.sub.3, 300 MHz) .delta. 12.11 (s, 1H),
8.02 (d, J=9.9 Hz, 1H), 7.16 (d, J=6.1 Hz, 2H), 6.79 (m, 1H), 6.20
(s, 1H), 5.18 (s, 1H), 4.80 (m, 1H), 1.75 (m, 1H), 1.41 (d, J=6.6
Hz, 3H), 0.94 (m, 2H), 0.68 (m, 2H).
Example 100
##STR00116##
[0428]
(2S,2'S)-2-[2-(3,5-Difluorophenyl)-2-hydroxyacetylamino]-N-(5-phene-
thyl-2H-pyrazol-3-yl)-propionamide
[0429] MS 429 (M+H), 451 (M+Na).
Example 101
##STR00117##
[0430]
(2S,2'S)--N-{5-[1-(4-Chlorophenylsulfanyl)-1-methylethyl]-2H-pyrazo-
l-3-yl}-2-[2-(3,5-difluorophenyl)-2-hydroxyacetylamino]-propionamide
[0431] MS 509 (M+H), 531 (M+Na).
[0432] Anal Calcd for
C.sub.23H.sub.23ClF.sub.2N.sub.4O.sub.3S.H.sub.2O: C, 52.42; H,
4.78; N, 10.63. Found: C, 52.55; H, 4.90; N, 10.40.
Example 102
##STR00118##
[0433]
(2S,2'S)-2-Hydroxy-3-methyl-N-{1-[5-(1-methyl-1-phenylethyl)-2H-pyr-
azol-3-ylcarbamoyl]ethyl}-butyramide
[0434] MS 373 (M+H).
Example 103
##STR00119##
[0435]
(2S)-2-Acetylamino-N-[5-(1,1-dimethyl-2-phenylethyl)-2H-pyrazol-3-y-
l]-propionamide
[0436] MS 330 (M+H), 351 (M+Na).
[0437] .sup.1H NMR (DMSO-d.sub.6, 300 MHz) .delta. 12.07 (s, 1H),
10.23 (s, 1H), 8.05 (d, J=6.6 Hz, 1H), 7.15 (m, 3H), 6.88 (d, J=7.1
Hz, 2H), 6.17 (s, 1H), 4.36 (m, 1H), 2.83 (s, 2H), 1.81 (s, 3H),
1.19 (m, 9H).
Example 104
##STR00120##
[0438]
(2S)-2-[2'-(3,5-Difluorophenyl)-2'-oxo-acetylamino]-N-[5-(1-methyl--
4-phenylethyl)-2H-pyrazol-3-yl]-propionamide
[0439] MS 441 (M+H).
[0440] Anal Calcd for C.sub.23H.sub.22F.sub.2N.sub.4O.sub.3: C,
62.72; H, 5.03; N, 12.72. Found: C, 62.36; H, 5.00; N, 12.41.
[0441] .sup.1H NMR (CDCl.sub.3, 300 MHz) .delta. 11.63 (s, 1H),
11.06 (s, 1H), 8.27 (d, J=9.3 Hz, 1H), 7.83 (m, 2H), 7.31-7.21 (m,
4H), 7.16-7.05 (m, 2H), 6.70 (s, 1H), 4.90 (m, 1H), 1.74 (s, 6H),
1.53 (d, J=7.1 Hz, 3H).
Example 105
##STR00121##
[0442]
(2S,2'R)-2-[2'-(3,5-Difluorophenyl)-2'-hydroxyacetylamino]-N-[5-(1--
methyl-1-phenylethyl)-2H-pyrazol-3-yl]-propionamide
[0443] MS 443 (M+H), 465 (M+Na).
[0444] Anal Calcd for C.sub.23H.sub.24F.sub.2N.sub.4O.sub.3.0.5
H.sub.2O: C, 61.19; H, 5.58; N, 12.41. Found: C, 61.72; H, 5.54; N,
12.34.
Example 106
##STR00122##
[0445]
(2S,2'S)-2-[2'-(3,5-Difluorophenyl)-2'-hydroxyacetylamino]-N-[5-[1--
2,3-difluorophenyl)-1-methylethyl]-2H-pyrazol-3-yl]-propionamide
[0446] MS 479 (M+H), 501 (M+Na).
[0447] Anal Calcd for C.sub.23H.sub.22F.sub.4N.sub.4O.sub.3.0.75
H.sub.2O: C, 56.15; H, 4.81; N, 11.39. Found: C, 56.23; H, 4.91; N,
11.01.
Example 107
##STR00123##
[0448]
N-[5-(1-Cyclohexyl-1-methyl-ethyl)-2H-pyrazol-3-yl]-2-[2-(3,5-diflu-
oro-phenyl)-2-hydroxy-acetylamino]-propionamide
[0449] MS 449.2 (M+H), 471.3 (M+Na).
[0450] .sup.1H NMR (CD.sub.3OD, 300 MHz) .delta. 7.09 (m, 2H), 6.87
(m, 1H), 6.32 (s, 1H), 5.08 (s, 1H), 1.40-1.80 (m, 5H), 1.41 (d,
J=7.2 Hz, 3H), 1.24 (s, 6H), 0.90-1.35 (m, 6H).
[0451] .sup.13C NMR (CD.sub.3OD, 75 MHz) .delta. 175.1, 173.4,
165.4 (dd, J=12.3, 245.7 Hz), 155.1, 147.0 (t, J=9.5 Hz), 122.1,
111.5 (dd, J=8.3, 17.6 Hz), 104.8 (t, J=25.9 Hz), 96.8, 74.9, 51.1,
50.4, 39.3, 29.8, 28.9, 28.5, 25.8, 19.4.
Example 108
Synthesis of
(2S,2'S)-2-[2'-(3,5-Difluorophenyl)-2'-hydroxyacetylamino]-N-[5-(1-methyl-
-4-phenylpiperidin-4-yl)-2H-pyrazol-3-yl]-propionamide
##STR00124## ##STR00125##
[0452]
(2S)-1-[2-tert-Butyl-5-(1-methyl-4-phenylpiperidin-4-yl)-2H-pyrazol-
-3-ylcarbamoyl]-ethylcarbamic acid benzyl ester (6)
[0453] A solution of 0.67 g (1.3 mmol) of 5 in 10 mL of absolute
MeOH was stirred at RT as 0.33 mL of a 37% aq. solution of
formaldehyde was added followed by 0.33 g (4.0 mmol) of NaOAc and
0.27 g (4.3 mmol) of NaCNBH.sub.3. After 16 h, the solvent was
removed by rotary evaporation and the residue was extracted with
water and EtOAc. The organic layer was dried over MgSO.sub.4,
filtered and the solvent was removed by rotary evaporation.
Purification of the material on silica gel using 4% Et.sub.3N-MeOH
as eluant afforded 0.41 g (60%) of ester 6 as a clear, colorless
glassy solid.
[0454] MS 518 (M+H).
##STR00126##
(2S)-2-Amino-N-[2-tert-butyl-5-(1-methyl-4-phenyl-piperidin-4-yl)-2H-pyra-
zol-3-yl]-propionamide (7)
[0455] A mixture of 0.36 g (0.7 mmol) of ester 6 and 0.36 g of 10%
palladium on carbon in 5.0 mL of absolute EtOH was shaken under 30
psi of H.sub.2. After 16 h, the mixture was filtered through Celite
and 0.36 g of fresh 10% palladium on carbon was added to the
filtrate. The mixture was again shaken under 30 psi of H.sub.2 and
this procedure was repeated until consumption of all starting
material. The reaction mixture was filtered through Celite and the
filtrate was concentrated by rotary evaporation. Purification of
the material on silica gel using 4% Et.sub.3N-MeOH as eluant
afforded 0.14 g (51%) of amine 7 as clear, colorless oil.
##STR00127##
(2S,2'S)-2-[2'-(3,5-Difluorophenyl)-2'-hydroxyacetylamino]-N-[5-(1-methyl-
-4-phenylpiperidin-4-yl)-2H-pyrazol-3-yl]-propionamide
[0456] MS 498 (M+H).
Example 109
Synthesis of
(2S,2'S)--N-[5-(1-Cyclopropyl-4-phenylpiperidin-4-yl)-2H-pyrazol-3-yl]-2--
[2'-(3,5-difluorophenyl)-2'-hydroxylacetylamino]-propionamide
##STR00128## ##STR00129##
[0457]
(S)-1-[2-tert-Butyl-5-(1-cyclopropyl-4-phenyl-piperidin-4-yl)-2H-py-
razol-3-ylcarbamoyl]-ethylcarbamic acid benzyl ester (11)
[0458] A solution of 1.22 g (2.4 mmol) of 5 in 20 mL of absolute
MeOH was stirred at rt as 1.42 g (2.4 mmol) of conc. acetic acid,
1.28 g of 4 .ANG. molecular sieves and 2.15 g (12.3 mmol) of ketal
10 were added followed by 0.68 g (10.8 mmol) of NaCNBH.sub.3. The
reaction mixture was refluxed for 16 h, cooled to RT, and filtered
to remove solids. The filtrate was concentrated by rotary
evaporation and extracted with water and Et0Ac. The organic layer
was dried over MgSO.sub.4, filtered and the solvent was removed by
rotary evaporation to afford 0.97 g (74%) of 11 as a clear,
colorless, glassy solid.
##STR00130##
(2S,2'S)--N-[5-(1-Cyclopropyl-4-phenylpiperidin-4-yl)-2H-pyrazol-3-yl]-2--
[2'-(3,5-difluorophenyl)-2'-hydroxylacetylamino]-propionamide
[0459] MS 524 (M+H), 546 (M+Na).
Example 110
Synthesis of
N-(5-tert-Butyl-2H-pyrazol-3-yl)-3-[(3,5-difluoro-phenyl)-formyl-amino]-b-
enzamide
##STR00131##
[0461] A solution of 4.0 mmoles (1.0 eq.) of
3,5-difluorobenzaldehyde and 4.0 mmole of 2-aminobenzoic acid in 60
mL of MeOH was prepared. While stirring at RT, 8.0 mmole (2.0 eq.)
of NaOAc was added followed by 9.0 mmole of NaCNBH.sub.3. The
reaction solution was stirred at RT for 1.5 hr and rotary
evaporated. The residue was taken up in 30 mL H.sub.2O and 50 mL
EtOAc. The layers were separated. The aqueous layer was extracted
with EtOAc (2.times.50 mL). The combined organic layers were dried
over Na.sub.2SO.sub.4 and vacuum filtered. The filtrate was rotary
evaporated and dried under vacuum to give coupled product 5
(78%).
##STR00132##
[0462] A solution of 3.1 mmoles (1.0 eq.) of 5 and 3.5 mmoles (1.1
eq.) of aminopyrazole in 10 mL of dry pyridine was cooled to
-10.degree. C. To the cooled solution, 3.4 mmoles (1.1 eq.) of
POCl.sub.3 was added. After stirring for 15 min, the orange
reaction mixture was concentrated. The residue was taken up in 30
mL of H.sub.2O, 20 mL of 2 M HCl, and 70 mL of EtOAc. The layers
were separated and the aqueous layer was extracted with EtOAc
(2.times.70 mL). The combined organic extracts were washed with
saturated aqueous NaHCO.sub.3, followed by drying over
Na.sub.2SO.sub.4 and vacuum filtering. The filtrate was rotary
evaporated. The crude product was flash chromatographed on silica
using 20% EtOAc/hexanes as eluant to afford product 6 (25%).
##STR00133##
N-(5-tert-Butyl-2H-pyrazol-3-yl)-3-[(3,5-difluoro-phenyl)-formyl-amino]-b-
enzamide
[0463] A solution of 0.40 mmole of 6 was refluxed in 5 mL of formic
acid for 15 min. After cooling to RT, the solution was added
dropwise to saturated aqueous NaHCO.sub.3. After ensuring pH 8, the
solution was extracted with EtOAc (3.times.70 mL). The combined
organic extracts were dried over Na.sub.2SO.sub.4 and vacuum
filtered. The filtrate was rotary evaporated. The crude material
was flash chromatographed on silica using 75% EtOAc/hexanes as
eluant to afford product 7 (57%). This compound does not fall
within general Formula I; however, it is a compound of interest
because it inhibits .beta.-amyloid peptide release and/or its
synthesis. MS 413.2 (M+H), 435.2 (M+Na).
[0464] .sup.1H NMR (CDCl.sub.3, 300 MHz) .delta. 11.07 (s(br), 1H),
10.16 (s, 1H), 8.86 (s, 1H), 8.16 (s, 1H), 7.79 (d , J=7.7 Hz, 1H),
7.36 (m, 1H), 7.19 (m, 1H), 6.65 (m, 4H), 4.96 (s, 2H), 1.29 (s,
9H).
Example 111
N-(5-tert-Butyl-2H-pyrazol-3-yl)-3-(3,5-difluoro-phenylamino)-benzamide
##STR00134##
[0466] A suspension of 7.6 mmoles (1.0 eq.) of 2-aminobenzoic acid,
8.0 mmoles (1.1 eq.) of 3,5-difluorobromobenzene, 0.16 mmole (0.02
eq.) of Pd(OAc).sub.2, 0.16 mmole (0.02 eq.) of (S)-BINAP, and 31.8
mmoles (4.0 eq.) of sodium tert-butoxide in 20 mL of dry toluene
was heated at 90.degree. C. for 15.5 hrs. The mixture was cooled to
RT and 40 mL of saturated aqueous NH.sub.4Cl was added. The layers
were separated and the aqueous layer was extracted with EtOAc
(3.times.70 mL). The combined organic layers were dried over
Na.sub.2SO.sub.4 and vacuum filtered. The filtrate was rotary
evaporated. The crude material was flash chromatographed on silica
with 75% EtOAc/hexanes as eluant to give coupled product 8
(15%).
##STR00135##
[0467] A solution of 1.4 mmoles (1.0 eq.) of 8 and 1.5 mmoles (1.1
eq.) of aminopyrazole in 5 mL of dry pyridine was cooled to
-10.degree. C. To the cooled solution, 1.5 mmoles (1.1 eq.) of
POCl.sub.3 was added. After stirring for 20 min, the orange
reaction mixture was concentrated. The residue was taken up in 20
mL of H.sub.2O, 10 mL of 2 M HCl, and 50 mL of EtOAc. The layers
were separated and the aqueous layer was extracted with EtOAc
(2.times.50 mL). The combined organic extracts were washed with
saturated aqueous NaHCO.sub.3 followed by drying over
Na.sub.2SO.sub.4 and vacuum filtering. The filtrate was rotary
evaporated. The crude product was flash chromatographed on silica
using 20% EtOAc/hexanes as eluant to afford product 9 (30%).
##STR00136##
N-(5-tert-Butyl-2H-pyrazol-3-yl)-3-(3,5-difluoro-phenylamino)-benzamide
[0468] Hydrogen chloride gas was bubbled through a solution of 0.29
mmole of 9 in 15 mL of MeOH for 2 min. The reaction solution was
refluxed for 5 hrs. The solution was rotary evaporated. The residue
was dissolved in 40 mL of saturated aqueous NaHCO.sub.3 and
extracted with EtOAc (3.times.50 mL). The combined organic extracts
were dried over Na.sub.2SO.sub.4 and vacuum filtered. The filtrate
was rotary evaporated. The crude product was flash chromatographed
on silica with a step gradient of 50% and 75% EtOAc/hexanes as
eluants to afford product 91891 (12%). This compound does not fall
within general Formula I, however, it also inhibits .beta.-amyloid
peptide release and/or its synthesis. .sup.1H NMR (CD.sub.3OD, 300
MHz) .delta. 7.71 (s(br), 1H), 7.55 (d, J=7.7 Hz, 1H), 7.46 (m,
1H), 7.37 (d, J=7.7 Hz, 1H), 6.67 (d, J=7.7 Hz, 2H), 6.47 (s, 1H),
6.38 (m, 1H), 1.38 (s, 9H).
Example 112
Synthesis of Product 14
##STR00137##
[0470] A solution of 1.2 mmole of 13 in 15 mL of TFA was refluxed
overnight. After 17 hrs, the solution was cooled to RT and added
dropwise to saturated aqueous NaHCO.sub.3. After ensuring pH 8, the
solution was extracted with EtOAc (3.times.70 mL). The combined
organic extracts were dried over MgSO.sub.4 and vacuum filtered.
The filtrate was rotary evaporated and dried under vacuum to afford
the product 14.
##STR00138##
[0471] The n-butyl isocyanate intermediate was generated by
treating 0.96 mmole (1.0 eq.) of 1-aminobutane with 1.9 mmole (2.0
eq.) of 1.93 M phosgene in toluene. The 1-aminobutane was dissolved
in 5 mL of CH.sub.2Cl.sub.2 and 5 mL of saturated aqueous
NaHCO.sub.3 and cooled to 0.degree. C. The phosgene was added to
the methylene chloride layer and stirred vigorously for 10 min. The
layers were separated. The organic layer was dried over MgSO.sub.4
and vacuum filtered. The filtrate was rotary evaporated and dried
under vacuum to afford n-butyl isocyanate.
[0472] In a separate flask, a solution of 0.95 mmole (1.0 eq.) of
14, 1.2 mmole (1.3 eq.) of Et.sub.3N in 5 mL of THF was prepared. A
solution of the n-butyl isocyanate in 2 mL of CH.sub.2Cl.sub.2 was
added and stirred at rt for 35 min. The reaction mixture was rotary
evaporated. The crude product was purified on a TLC preparatory
plate using 7% MeOH/CH.sub.2Cl.sub.2 as eluant to give urea product
104228. MS 408.2 (M+H), 429.8 (M+Na). .sup.1H NMR (CDCl.sub.3, 300
MHz) .delta. 11.96 (s(br), 1H), 6.80 (d, J=7.1 Hz, 2H), 6.59 (m,
2H), 5.63 (d, J=8.2 Hz, 1H), 4.79 (m, 2H), 3.01 (m, 2H), 1.73 (s,
3H), 1.71 (s, 3H), 1.41 (d, J=7.1 Hz, 3H), 1.33 (m, 2H), 1.21 (m,
2H), 0.82 (t, J=7.1 Hz, 3H).
Example 113
##STR00139##
[0473]
(2S)-2-(3,4-Dichlorophenylamino)-N-{5-[1-(3,5-difluorophenyl)-1-met-
hylethyl]-2H-pyrazol-3yl}-propionamide
[0474] MS 453 (M+H).
[0475] Anal Calcd for C.sub.21H.sub.20Cl.sub.2F.sub.2N.sub.4O.0.5
H.sub.2O: C, 54.56; H, 4.58; N, 12.12. Found: C, 54.76; H, 4.51; N,
11.83.
Example 114
Synthesis of
5-(2-[(3,5-Difluorophenyl)acetylamino]-2-(isopropylthio)acetylamino)-3-te-
rt-butylpyrazole 18
##STR00140##
[0476] (3,5-Difluorophenyl)acetamide 1
##STR00141##
[0478] A solution of 10 g of (3,5-difluorophenyl)acetic acid in 100
mL of tetrahydrofuran was treated with 9.9 g of
carbonyldiimidazole. The mixture was stirred at 25.degree. C. for 3
hours then a vigorous stream of anhydrous ammonia was bubbled
through the solution for 1 hour. The mixture was stirred for 2 days
and then the solvent was evaporated at reduced pressure. The
residue was suspended in 100 mL of distilled water. The mixture was
filtered and the filtrant was washed with distilled water twice.
After drying in a stream of air, the (3,5-difluorophenyl)acetamide
was obtained as 8.4 g of an ivory solid. .sup.1H-NMR
(DMSO-d.sub.6): .delta. 7.51 (broad s, 1H), 7.15-6.85 (m, 3H), 3.40
(s, 2H). MS (EI): m/z 171. EA Calc'd for C--.sub.8H.sub.7NO: C,
56.13; H, 4.13; N, 8.18. Found: C, 56.16; H, 4.05; N, 8.07.
2-[(3,5-Difluorophenyl)acetylamino]-2-hydroxyacetic acid 2
##STR00142##
[0480] A mixture of 7.2 g of (3,5-difluorophenyl)acetamide 1 and
4.6 g of glyoxylic acid hydrate in 75 mL of acetone was refluxed 24
hours. The solvent was evaporated and the residue was dissolved in
300 mL of ethyl acetate. This solution was washed with 150 mL of
distilled water. The aqueous phase was discarded. The organic phase
was extracted with a solution prepared by diluting 50 mL of a
saturated aqueous solution of sodium bicarbonate with 150 mL of
distilled water. The organic phase was discarded and the aqueous
bicarbonate solution was acidified with 80 mL of 1 N aqueous sodium
bisulfate solution. The acidified solution was extracted with three
successive 125 mL portions of ethyl acetate. The combined organic
extracts were washed with brine and then dried over magnesium
sulfate. The solution was filtered and the solvent was evaporated
to yield 2-[(3,5-difluorophenyl)acetylamino]-2-hydroxyacetic acid
as 5.2 g of a white solid. .sup.1H-NMR (DMSO-d.sub.6): .delta. 8.95
(d, J=8 Hz, 1H), 7.15-6.90 (m, 3H), 5.37 (d, J=8 Hz, 1H), 3.50 (s,
2H). .sup.13C-NMR (DMSO-d6.sub.6) .delta. 171.5, 169.4, 162.4 (dd,
J.sub.C-F=13, 244 Hz), 140.6 (t, J.sub.C-F=10 Hz), 112.6 (dd,
J.sub.C-F=8, 17 Hz), 71.5, 41.7. MS (CI+): m/z 246. EA Calc'd for
C--.sub.10H.sub.9F.sub.2NO.sub.4: C, 48.98; H, 3.71; N, 5.71.
Found: C, 49.02; H, 3.72; N, 5.51.
Methyl 2-[(3,5-difluorophenyl)acetylamino]-2-methoxyacetate 2A
##STR00143##
[0482] A 0.degree. C. mixture of 1.0 g of
2-[(3,5-Difluorophenyl)acetylamino]-2-hydroxyacetic acid 2, 20 mL
of methanol, and 6 mL of dichloromethane was treated with 0.3 mL of
sulfuric acid. The mixture was stirred 2 days at 25.degree. C. The
solvent was evaporated at reduced pressure and the residue was
partitioned between distilled water and ethyl acetate. The aqueous
phase was washed with ethyl acetate, and the combined organic
extracts were washed successively with a saturated aqueous solution
of sodium bicarbonate and brine. The solution was dried over
magnesium sulfate and filtered. The solvent was evaporated at
reduced pressure. The residue was chromatographed on silica gel
eluting with 30% ethyl acetate in hexanes. Fractions containing the
major component of the mixture were combined to yield methyl
2-[(3,5-difluorophenyl)acetylamino]-2-methoxyacetate 2A as 0.52 g
of a white powder. .sup.1H-NMR (DMSO-d.sub.6): .delta. 9.15 (d,
J=8.7 Hz, 1H), 7.15-6.85 (m, 3H), 5.34 (d, J=8.7 Hz, 1H), 3.69 (s,
3H), 3.60 (s, 3H), 3.24 (s, 2H).). .sup.13C-NMR (CDCl.sub.3)
.delta. 169.9, 168.17, 163.0 (dd, J.sub.C-F=12.8, 248 Hz), 137.4
(t, J.sub.C-F=9 Hz), 112.2 (dd, J.sub.C-F=8, 17 Hz), 102.9 (t,
J.sub.C-F=25 Hz), 78.3, 56.8, 52.8, 42.7. MS (CI+): m/z 274. EA
Calc'd for C.sub.12H.sub.13F.sub.2NO.sub.4: C, 52.74; H, 4.81; N,
5.13. Found: C, 52.52; H, 4.87; N, 5.03.
2-[(3,5-Difluorophenyl)acetylamino]-2-methoxyacetic acid 3
##STR00144##
[0484] Methyl 2-[(3,5-difluorophenyl)acetylamino]-2-methoxyacetate
2A (0.4 g) was dissolved in 10 mL of dioxane and treated with 2.8
mL of a 1.0 N aqueous sodium hydroxide solution. After 1.5 h most
of the solvent was removed by evaporation at reduced pressure. The
mixture was acidified with an aqueous solution of sodium bisulfate
and washed with two successive portions of ether. The combined
organic extracts were washed with brine and then dried with
magnesium sulfate. The solution was filtered and the solvent was
evaporated at reduced pressure to yield
2-[(3,5-Difluorophenyl)acetylamino]-2-methoxyacetic acid 3 as 0.35
g of a white powder. .sup.1H-NMR (DMSO-d.sub.6): .delta. 9.06 (d,
J=8.7 Hz, 1H), 7.15-6.85 (m, 3H), 5.23 (d, J=8.7 Hz, 1H), 3.57 (s,
3H), 3.21 (s, 2H).
5-Amino-1,3-di(tert-butyl)pyrazole (2363-46) 17
##STR00145##
[0486] A mixture of 12.5 g of 4,4-dimethyl-3-oxopentanenitrile, 25
g of tert-butylhydrazine hydrochloride, and 27.6 g of potassium
carbonate in 400 mL of ethanol was refluxed for 24 h. The mixture
was cooled to 25.degree. C. and then it was filtered. The solvent
was evaporated from the filtrate and the residue was partitioned
between ethyl acetate and distilled water. The organic phase was
washed with brine and dried (MgSO.sub.4). The solvent was
evaporated at reduced pressure and the residue was dissolved in a
small volume of ether. The mixture was diluted with an equal volume
of hexanes and then the volume of the solution was reduced by one
half by evaporation at reduced pressure. The white solid which
precipitated was collected by filtration to give 8.7 g of
5-Amino-1,3-di(tert-butyl)pyrazole 17. .sup.1H-NMR (CDCl.sub.3):
.delta. 5.43 (s, 1H), 1.62 (s, 9H), 1.25 (s, 9H).
5-(2-[(3,5-Difluorophenyl)acetylamino]-2-(isopropylthio)acetylamino)-3-ter-
t-butylpyrazole 18
##STR00146##
[0488] Phosphorus oxychloride (0.1 mL) was added to a 0.degree. C.
solution of 0.3 g of acid 14 and 0.19 g of
5-amino-1,3-di-tert-butylpyrazole 17 in 5 mL of pyridine. After 20
min the reaction mixture was poured onto ice. The resulting
suspension was washed with ethyl acetate twice. The combined
organic extracts were washed with distilled water, a 1 N aqueous
solution of sodium bisulfate, and brine. The solution was dried
(MgSO.sub.4) and the solvent was evaporated at reduced pressure.
The residue, which had solidified during the solvent evaporation
step, was slurried with a small amount of ethyl acetate. The
mixture was filtered and the solid thus obtained was dried in a
stream of air to give 75 mg of a white solid.
[0489] A 59 mg sample of this material was suspended in formic
acid. The reaction flask was then partially immersed in a preheated
115.degree. C. oil bath. After 20 minutes at reflux the solution
was cooled to 25.degree. C. The formic acid was removed by
evaporation at reduced pressure. After completely removing the
formic acid the residue was dissolved in a few mL of chloroform and
the resulting solution was filtered. The chloroform was evaporated
from the filtrate and the residue suspended in tert-butyl methyl
ether. The suspension was refluxed for 5 min and then it was cooled
to 25.degree. C. It was filtered and the solid thus obtained was
dried in a stream of air.
5-(2-[(3,5-Difluorophenyl)acetylamino]-2-(isopropylthio)acetylamino)-3-te-
rt-butylpyrazole 18 is obtained as 37 mg of a white powder.
[0490] .sup.1H-NMR (CDCl.sub.3): .delta. 7.06 (d, J=10 Hz, 1H),
6.9-6.7 (m, 3H), 6.54 (s, 1H), 5.99 (d, J=10 Hz, 1H), 3.62 (s, 2H),
3.1-3.1 (m, 1H), 1.31 (s, 9H), 1.27 (d, J=5.4 Hz, 3H), 1.23 (d,
J=6.6 Hz, 3H).). .sup.13C-NMR (DMSO-d.sub.6) .delta. 172.4, 168.4,
164.9 (dd, J.sub.C-F=13, 245 Hz), 156.0, 147.6, 141.3 (t,
J.sub.C-F=ca. 10 Hz), 113.7 (dd, J.sub.C-F=7.7, 17 Hz), 103.6 (t,
J.sub.C-F=26 Hz), 94.7, 56.7, 43.2, 37.0, 32.6, 30.8, 27.6, 24.8,
24.6. MS (CI+): m/z 447 (M+Na.sup.+). EA Calc'd for
C.sub.20H.sub.26F.sub.2N.sub.4O.sub.2S.1.7 H.sub.2O: C, 52.78; H,
6.51; N, 12.31. Found: C, 52.57; H, 6.16;N, 12.18.
Example 115
Synthesis of
5-(2-[(3,5-Difluorophenyl)acetylamino]-2-(2,2,2-(trifluoroethyl)thio)acet-
ylamino)-3-tert-butylpyrazole 19
##STR00147##
[0492] This compound was prepared from the acid 2 (Example 114)
using a three step procedure analogous to that used for the
preparation of 18 (Example 114). One difference is that the amide
bond formation was performed using PhOP(O)Cl.sub.2 in place of
POCl.sub.3. After this amide bond forming step, the intermediate
was purified by filtration through silica gel eluting with 20%
ethyl acetate in hexane. Compound 19 gave the following analytical
data: 48.92% C, 4.59% H, 11.80% N.
Example 116
Synthesis of
5-(2-[(3,5-Difluorophenyl)acetylamino]-2-(ethoxyacetypamino-3-tert-butylp-
yrazole 20
##STR00148##
[0494] A suspension of 0.4 grams of compound 18 (Example 114) in 20
mL of 1:1 dichloromethane/ethanol was treated with 0.375 grams of
N-bromosuccinimide. After one hour the solution was treated with 1
mL of a saturated aqueous solution of sodium bisulfite. After an
additional 5 min, the mixture was partitioned between water and
ethyl acetate. The organic phase was washed with brine and dried
with magnesium sulfate. The solvent was evaporated at reduced
pressure and the residue was recrystallized from aqueous methanol
twice. The resulting solid was dissolved in 20 mL of ethanol and
treated with 50 mg of 10% palladium on carbon and 300 mg of sodium
acetate. The mixture was agitated under 50 psi hydrogen gas for 14
h. The mixture was filtered through celite and then the solvent was
evaporated at reduced pressure. The residue was partitioned between
water and ethyl acetate. The organic phase was washed with brine
and dried over magnesium sulfate. The solvent was evaporated at
reduced pressure. The residue was recrystallized from a mixture of
tert-butyl methyl ether and hexanes. The product was isolated as a
white powder. Analysis gave 56.91% C, 6.11% H, and 14.20% N.
Example 117
Synthesis of
5-(2-[(3,5-Difluorophenyl)acetylamino]-2-(methylthio)acetylamino)-3-tert--
butylpyrazole 21
##STR00149##
[0496] This compound was prepared from the acid 11 (Example 114)
using a three step procedure analogous to that used for the
preparation of 18 from 14 (Example 114). Compound 21 gave the
following mass spectral data: m/z=419.2 (M+H).sup.+.
Example 118
Monoethyl 2-(tert-butoxycarbonylamino)malonate 22
##STR00150##
[0498] A commercial sample of diethyl 2-aminomalonate hydrochloride
(7.0 g) was treated with 5.5 mL of triethylamine in a mixture of 50
mL of tetrahydrofuran and 30 mL of dimethylformamide. The resulting
mixture was treated with 6.76 grams of di(tert-butyl)pyrocarbonate
and stirred at 25.degree. C. for 14 hours. The solvent was
evaporated at reduced pressure and the residue was partitioned
between ethyl acetate and water. The organic phase was washed with
a saturated aqueous solution of sodium bicarbonate, a 1 molar
solution of sodium bisulfate and then with brine. It was dried over
magnesium sulfate and then the solvent was evaporated at reduced
pressure. The resulting oil was dissolved in a mixture of 25 mL of
ethanol and 28 mL of acetone. The resulting solution was cooled to
0.degree. C. and treated with 26 mL of a 1.0 N aqueous solution of
sodium hydroxide. The mixture was allowed to warm to 25.degree. C.
and stirred 18 hours. The solvent was evaporated at reduced
pressure and the residue was partitioned between ethyl acetate and
water. The aqueous phase was acidified with aqueous sodium
bisulfate solution and extracted with ethyl acetate. The organic
phase was dried (MgSO.sub.4) and the solvent was evaporated to give
an oil. The properties of this compound were in good accord with
those described previously (Journal of Medicinal Chemistry 1982,
397).
Example 119
Synthesis of
1,3-bis(tert-butyl)-5-[2-(3,5-difluorophenyl)acetylamino-2-(ethoxycarbony-
l)acetylamino]pyrazole 23
##STR00151##
[0500] A mixture of 3.0 g of ester 22 (Example 118) and 2.6 grams
of aminopyrazole 17 (Example 114) in 50 mL of 0.degree. C. pyridine
was treated with 1.2 mL of phosphorus oxychloride. After 1 hour the
mixture was poured into 250 mL of ice water. The mixture was
filtered and the filtrant was washed with water several times. The
solid was dried in a stream of air to give an off white solid.
[0501] This material was dissolved in 100 mL of trifluoroacetic
acid and stirred for 1.5 hours. The solvent was evaporated and the
residue was partitioned between ethyl acetate and an excess of a
saturated aqueous solution of sodium bicarbonate. The aqueous phase
was washed with ethyl acetate and and the combined organic extracts
were washed with brine and dried over magnesium sulfate. The
solvent was evaporated at reduced pressure. The resulting oil was
dissolved in a minimum volume of ethyl acetate and allowed to stand
until seed crystals formed. The solvent was evaporated, and the
residue was dissolved in a minimum volume of ether. The solution
was diluted with a large excess of hexanes and the volume was
reduced by evaporation at reduced pressure. The resulting solid was
collected by filtration and washed with hexanes.
[0502] A solution of 1.0 gram of this solid, 0.43 g of
N-methylmorpholine and 0.53 g of HOBt in 30 mL of dimethylformamide
was treated with 0.75 grams of EDC hydrochloride. After 18 hours
the mixture was partitioned between ethyl acetate and water. The
aqueous phase was washed with water twice and the combined organic
extracts were washed twice with water, once with a 1 N aqueous
solution of sodium bisulfate, once with a saturated aqueous
solution of sodium bicarbonate, and finally with brine. The
solution was dried over magnesium sulfate and the solvent was
evaporated at reduced pressure. Treating the residual oil with
ether gave a solid precipitate, which was collected by
filtration.
Example 120
Synthesis of
3-tert-butyl-5-[2-(3,5-difluorophenyl)acetylamino-2-(ethoxycarbonyl)acety-
lamino]pyrazole 24
##STR00152##
[0504] 250 mg of compound 23 (Example 119) was refluxed in 3 mL of
formic acid for 20 min. The mixture was cooled to 25.degree. C. and
diluted with 12 mL of distilled water. The resulting precipitate
was collected by filtration and dried in a stream of air. It was
suspended in 5 mL of chloroform and filtered. The solvent was
evaporated from the filtrate at reduced pressure. The residue was
stirred with 5 mL of tert-butyl methyl ether for one hour, and then
recovered by filtration. The product was obtained as a white
powder. Analysis gave 56.84% C, 5.87% H, 13.28% N.
Example 121
Synthesis of
3-tert-butyl-5-[2-(3,5-difluorophenyl)acetylamino-2-(pyrrolidinocarbonyl)-
acetylamino]pyrazole 24
##STR00153##
[0506] 0.7 grams of the ester 23 (Example 119) was dissolved in a
mixture of 10 mL of ethanol and 10 mL of acetone. The resulting
solution was treated with 2.46 mL of 1.0 N aqueous sodium hydroxide
solution. After 48 hours the mixture was acidified with aqueous
sodium bisulfate solution. It was partitioned between ethyl acetate
and water. The aqueous phase was washed with ethyl acetate and the
combined organic extracts were washed with brine. The solution was
dried over magnesium sulfate and the solvent was evaporated at
reduced pressure to give an ivory solid.
[0507] A solution 0.3 g of this solid, 0.062 mL of pyrrolidine, and
0.19 gram of HOBt in 15 mL of DMF was treated with 0.14 grams of
EDC hydrochloride. After 20 hours the mixture was partitioned
between ethyl acetate and water. The aqueous phase was washed with
ethyl acetate and the combined organic extracts were washed with
aqueous sodium bisulfate, pH 9 phosphate buffer, and brine. The
solution was dried over magnesium sulfate and the solvent was
evaporated to give a white powder.
[0508] This powder was refluxed in 1 mL of formic acid for 20
minutes. The solvent was evaporated in a stream of nitrogen. The
residue was partitioned between ethyl acetate and water. The
aqueous phase was washed with ethyl acetate, and the combined
organic extracts were washed with brine. The organic phase was
dried over magnesium sulfate, and the solvent was evaporated at
reduced pressure. The residual solid was briefly stirred with 1 mL
of tert-butyl methyl ether and then filtered. This solid gave the
following mass spectral data: m/z--448.2 (M+H).sup.+.
[0509] The following non-limiting formulation examples (Examples
122-131) illustrate representative pharmaceutical compositions of
the present invention.
Example 122
[0510] Hard gelatin capsules containing the following ingredients
are prepared:
TABLE-US-00001 Quantity Ingredient (mg/capsule) Active Ingredient
30.0 Starch 305.0 Magnesium stearate 5.0
[0511] The above ingredients are mixed and filled into hard gelatin
capsules in 340 mg quantities.
Example 123
[0512] A tablet formula is prepared using the ingredients
below:
TABLE-US-00002 Quantity Ingredient (mg/tablet) Active Ingredient
25.0 Cellulose, microcrystalline 200.0 Colloidal silicon dioxide
10.0 Stearic acid 5.0
[0513] The components are blended and compressed to form tablets,
each weighing 240 mg.
Example 124
[0514] A dry powder inhaler formulation is prepared containing the
following components:
TABLE-US-00003 Ingredient Weight % Active Ingredient 5 Lactose
95
[0515] The active ingredient is mixed with the lactose and the
mixture is added to a dry powder inhaling appliance.
Example 125
[0516] Tablets, each containing 30 mg of active ingredient, are
prepared as follows:
TABLE-US-00004 Quantity Ingredient (mg/tablet) Active Ingredient
30.0 mg Starch 45.0 mg Microcrystalline cellulose 35.0 mg
Polyvinylpyrrolidone 4.0 mg (as 10% solution in sterile water)
Sodium carboxymethyl starch 4.5 mg Magnesium stearate 0.5 mg Talc
1.0 mg Total 120 mg
[0517] The active ingredient, starch and cellulose are passed
through a No. 20 mesh U.S. sieve and mixed thoroughly. The solution
of polyvinylpyrrolidone is mixed with the resultant powders, which
are then passed through a 16 mesh U.S. sieve. The granules so
produced are dried at 50-60.degree. C. and passed through a 16 mesh
U.S. sieve. The sodium carboxymethyl starch, magnesium stearate,
and talc, previously passed through a No. 30 mesh U.S. sieve, are
then added to the granules which, after mixing, are compressed on a
tablet machine to yield tablets each weighing 120 mg.
Example 126
[0518] Capsules, each containing 40 mg of medicament are made as
follows:
TABLE-US-00005 Quantity Ingredient (mg/capsule) Active Ingredient
40.0 mg Starch 109.0 mg Magnesium stearate 1.0 mg Total 150.0
mg
[0519] The active ingredient, starch, and magnesium stearate are
blended, passed through a No. 20 mesh U.S. sieve, and filled into
hard gelatin capsules in 150 mg quantities.
Example 127
[0520] Suppositories, each containing 25 mg of active ingredient
are made as follows:
TABLE-US-00006 Ingredient Amount Active Ingredient 25 mg Saturated
fatty acid glyceride 2,000 mg
[0521] The active ingredient is passed through a No. 60 mesh U.S.
sieve and suspended in the saturated fatty acid glycerides
previously melted using the minimum heat necessary. The mixture is
then poured into a suppository mold of nominal 2.0 g capacity and
allowed to cool.
Example 128
[0522] Suspensions, each containing 50 mg of medicament per 5.0 mL
dose are made as follows:
TABLE-US-00007 Ingredient Amount Active Ingredient 50.0 mg Xanthan
gum 4.0 mg Sodium carboxymethyl cellulose (11%) 50.0 mg
Microcrystalline cellulose (89%) Sucrose 1.75 g Sodium benzoate
10.0 mg Flavor and Color q.v. Purified water to 5.0 mL
[0523] The active ingredient, sucrose and xanthan gum are blended,
passed through a No. 10 mesh U.S. sieve, and then mixed with a
previously made solution of the microcrystalline cellulose and
sodium carboxymethyl cellulose in water. The sodium benzoate,
flavor, and color are diluted with some of the water and added with
stirring. Sufficient water is then added to produce the required
volume.
Example 129
TABLE-US-00008 [0524] Quantity Ingredient (mg/capsule) Active
Ingredient 15.0 mg Starch 407.0 mg Magnesium stearate 3.0 mg Total
425.0 mg
[0525] The active ingredient, starch, and magnesium stearate are
blended, passed through a No. 20 mesh U.S. sieve, and filled into
hard gelatin capsules in 425.0 mg quantities.
Example 130
[0526] A subcutaneous formulation may be prepared as follows:
TABLE-US-00009 Ingredient Quantity Active Ingredient 5.0 mg Corn
Oil 1.0 mL
Example 131
[0527] A topical formulation may be prepared as follows:
TABLE-US-00010 Ingredient Quantity Active Ingredient 1-10 g
Emulsifying Wax 30 g Liquid Paraffin 20 g White Soft Paraffin to
100 g
[0528] The white soft paraffin is heated until molten. The liquid
paraffin and emulsifying wax are incorporated and stirred until
dissolved. The active ingredient is added and stirring is continued
until dispersed. The mixture is then cooled until solid.
Example 132
.beta.-amyloid Precursor Protein Accumulation Assay
[0529] An assay to evaluate the accumulation of A.beta. is used to
detect potential inhibitors of secretase. The assay uses the N 9
cell line, characterized for expression of exogenous APP by
immunoblotting and immunoprecipitation.
[0530] The effect of test compounds on the accumulation of A.beta.
in the conditioned medium is tested by immunoprecipitation as
described in WO 01/19797. Briefly, N 9 cells are grown to
confluency in the 6-well plates. Test compounds dissolved in DMSO
are added together with the addition of radiolabel. The cells are
incubated for 4 h at 37.degree. C. in a tissue culture
incubator.
[0531] At the end of the incubation period, the conditioned medium
is harvested and pre-cleared by the addition of normal mouse serum
and of protein A Sepharose, mixed by end-over-end rotation for 30
minutes at 4.degree. C., followed by a brief centrifugation in a
microfuge. The supernatant is then harvested and transferred to
fresh tubes containing of a monoclonal antibody directed against an
internal peptide sequence in A.beta. and protein A Sepharose. After
incubation overnight at 4.degree. C., the samples are washed. The
pellet after the last wash is resuspended in SDS sample buffer and
boiled for 3 minutes. The supernatant is then fractionated on SDS
gels. The gels are dried and exposed to X-ray film or analyzed by
phosphorimaging. The resulting image is analyzed for the presence
of A.beta. polypeptides. The steady-state level of A.beta. in the
presence of a test compound is compared to wells treated with DMSO
(1%) alone. A typical test compound is considered active if it
blocks A.beta. accumulation in the conditioned medium, and has and
IC.sub.50 less than 100 .mu.M.
Example 133
C-Terminus .beta.-Amyloid Precursor Protein Accumulation Assay
[0532] The effect of test compounds on the accumulation of
C-terminal fragments is determined by immunoprecipitation of APP
and fragments thereof from cell lysates as described in WO
01/19797. N 9 cells are metabolically labeled as above in the
presence or absence of test compounds. At the end of the incubation
period, the conditioned medium are harvested and cells lysed in
buffer. Lysates are precleared with normal rabbit serum and protein
A-Sepharose, followed by the addition of BC-1 antiserum and protein
A-Sepharose for 16 hours at 4.degree. C. The immunoprecipitates are
washed bound proteins eluted by boiling in SDS sample buffer and
fractionated by Tris/Tricine SDS-PAGE. After exposure to X-ray film
or phosphorimager, the resulting images are analyzed for the
presence of C-terminal APP fragments. The steady-state level of
C-terminal APP fragments is compared to wells treated with DMSO
(1%) alone. A typical test compound is considered active if it
stimulates C-terminal fragment accumulation in the cell lysates,
and has an IC.sub.50 less than 100 .mu.M.
Example 134
A.beta. Immunoprecipitation Assay
[0533] This immunoprecipitation assay as described in WO 01/19797
is specific for .gamma. secretase (i.e., proteolytic activity
required to generate the C-terminal end of A.beta. either by direct
cleavage or generating a C-terminal extended species which
subsequently further proteolyzed). Briefly, N 9 cells are
pulse-labeled in the presence of a reported .gamma. secretase
inhibitor for 1 h, followed by washing to remove the radiolabel and
the inhibitor. The media is replaced and test compounds are added.
A.beta. is isolated from the conditioned medium and C-terminal
fragments from cell lysates. The test compounds are characterized
to determine whether a stabilization of C-terminal fragments is
observed and whether A.beta. is generated from these accumulated
precursor. A typical test compound is considered active if it
prevents the generation of A.beta. out of accumulated C-terminal
fragments and has an IC.sub.50 less than 100 .mu.M.
Example 135
Cellular Screen for the Detection of Inhibitors of .beta.-Amyloid
Production
[0534] Compounds of Formula I are assayed for their ability to
inhibit .beta.-amyloid peptide production in a cell line possessing
the Swedish mutation. This screening assay employed cells
(K293=human kidney cell line) which were stably transfected with
the gene for amyloid precursor protein 751 (APP751) containing the
double mutation Lys.sub.651 Met.sub.652 to Asn.sub.651 Leu.sub.652
(APP751 numbering) in the manner described in WO 94/10569 and
Citron et al (Nature, 360:672-674 (1992)). This mutation is
commonly called the Swedish mutation. The cells, designated as "293
751 SWE", are plated in Corning 96-well plates at
1.5-2.5.times.10.sup.4 cells per well in Dulbecco's minimal
essential media (Sigma, St. Louis, Mo.) plus 10% fetal bovine
serum. Cell number is important in order to achieve .beta.-amyloid
ELISA results within the linear range of the assay (about 0.2 to
2.5 ng per mL).
[0535] Following overnight incubation at 37.degree. C. in an
incubator equilibrated with 10% carbon dioxide, media were removed
and replaced with 200 .mu.L of a compound of formula I (drug)
containing media per well for a two hour pretreatment period and
cells were incubated as above. Drug stocks were prepared in 100%
dimethyl sulfoxide such that at the final drug concentration used
in the treatment, the concentration of dimethyl sulfoxide did not
exceed 0.5% and, in fact, usually equaled 0.1%.
[0536] At the end of the pretreatment period, the media are again
removed and replaced with fresh drug containing media as above and
cells are incubated for an additional two hours. After treatment,
plates are centrifuged in a Beckman GPR at 1200 rpm for five
minutes at room temperature to pellet cellular debris from the
conditioned media. From each well, 100 .mu.L of conditioned media
or appropriate dilutions thereof are transferred into an ELISA
plate precoated with antibody 266 (Nature, 359:325-327 (1992))
against amino acids 13-28 of .beta.-amyloid peptide as described in
WO 94/10569 and stored at 4.degree. C. overnight. An ELISA assay
employing labelled antibody 6C6 (Nature, 359:325-327 (1992))
against amino acids 1-16 of .beta.-amyloid peptide is run the next
day to measure the amount of .beta.-amyloid peptide produced.
[0537] Cytotoxic effects of the compounds are measured by a
modification of the method of Hansen, et al. (J. Immun. Meth.,
119:203-210 (1989)). To the cells remaining in the tissue culture
plate is added 25 .mu.L of a
3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT)
(Sigma, St. Louis, Mo.) stock solution (5 mg/mL) to a final
concentration of 1 mg/mL. Cells are incubated at 37.degree. C. for
one hour, and cellular activity was stopped by the addition of an
equal volume of MTT lysis buffer (20% w/v sodium dodecylsulfate in
50% dimethylformamide, pH 4.7). Complete extraction is achieved by
overnight shaking at room temperature. The difference in the
OD.sub.562 nm and the OD.sub.650 mn is measured in a Molecular
Device's UV.sub.max microplate reader as an indicator of the
cellular viability.
[0538] The results of the .beta.-amyloid peptide ELISA are fit to a
standard curve and expressed as ng/mL .beta.-amyloid peptide. In
order to normalize for cytotoxicity, these results are divided by
the MTT results and expressed as a percentage of the results from a
drug free control.
Example 136
In Vivo Suppression of .beta.-Amyloid Release and/or Synthesis
[0539] This example illustrates how the compounds of this invention
could be tested for in vivo suppression of .beta.-amyloid release
and/or synthesis. For these experiments, 3 to 4 month old PDAPP
mice are used (Games et al., Nature 373:523-52 (1995)). Depending
upon which compound is being tested, the compound is usually
formulated at either 5 or 10 mg/ml. Because of the low solubility
factors of the compounds, they may be formulated with various
vehicles, such as corn oil (Safeway, South San Francisco, Calif.);
10% EtOH in corn oil (Safeway); 2-hydroxypropyl-.beta.-cyclodextrin
(Research Biochemicals International, Natick M A); and
carboxy-methyl-cellulose (Sigma Chemical Co., St. Louis, Mo.).
[0540] The mice are dosed subcutaneously with a 26 gauge needle and
3 hours later the animals are euthanized via CO.sub.2 narcosis and
blood is taken by cardiac puncture using a 1 cc 25G 5/8''
tuberculin syringe/needle coated with solution of 0.5 M EDTA, pH
8.0. The blood is placed in a Becton-Dickinson vacutainer tube
containing EDTA and spun down for 15 minutes at 1500.times.g at
5.degree. C. The brains of the mice are then removed and the cortex
and hippocampus are dissected out and placed on ice.
1. Brain Assay
[0541] To prepare hippocampal and cortical tissue for enzyme-linked
immunosorbent assays (ELISAs) each brain region is homogenized in
10 volumes of ice cold guanidine buffer (5.0 M guanidine-HCl, 50 mM
Tris-HCl, pH 8.0) using a Kontes motorized pestle (Fisher,
Pittsburgh Pa.). The homogenates are gently rocked on a rotating
platform for three to four hours at room temperature and stored at
-20.degree. C. prior to quantitation of .beta.-amyloid.
[0542] The brain homogenates are diluted 1:10 with ice-cold casein
buffer (0.25% casein, phosphate buffered saline (PBS), 0.05% sodium
azide, 20 .mu.g/ml aprotinin, 5 mM EDTA, pH 8.0, 10 .mu.g/ml
leupeptin), thereby reducing the final concentration of guanidine
to 0.5 M, before centrifugation at 16,000.times.g for 20 minutes at
4.degree. C. The .beta.-amyloid standards (1-40 or 1-42 amino
acids) were prepared such that the final composition equaled 0.5 M
guanidine in the presence of 0.1% bovine serum albumin (BSA).
[0543] The total .beta.-amyloid sandwich ELISA, quantitating both
.beta.-amyloid (aa 1-40) and .beta.-amyloid (aa 1-42) consists of
two monoclonal antibodies (mAb) to .beta.-amyloid. The capture
antibody, 266, is specific to amino acids 13-28 of .beta.-amyloid.
The antibody 3D6, which is specific to amino acids 1-5 of
.beta.-amyloid, is biotinylated and served as the reporter antibody
in the assay. The 3D6 biotinylation procedure employs the
manufacturer's (Pierce, Rockford Ill.) protocol for NHS-biotin
labeling of immunoglobulins except that 100 mM sodium bicarbonate,
pH 8.5 buffer is used. The 3D6 antibody does not recognize secreted
amyloid precursor protein (APP) or full-length APP but detects only
.beta.-amyloid species with an amino terminal aspartic acid. The
assay has a lower limit of sensitivity of about 50 pg/ml (11 pM)
and shows no cross-reactivity to the endogenous murine
.beta.-amyloid peptide at concentrations up to 1 ng/ml.
[0544] The configuration of the sandwich ELISA quantitating the
level of .beta.-amyloid (aa 1-42) employs the mAb 21F12 (which
recognizes amino acids 33-42 of .beta.-amyloid) as the capture
antibody. Biotinylated 3D6 is also the reporter antibody in this
assay which has a lower limit of sensitivity of about 125 pg/ml (28
pM).
[0545] The 266 and 21F12 capture mAbs are coated at 10 .mu.g/ml
into 96 well immunoassay plates (Costar, Cambridge Mass.) overnight
at room temperature. The plates are then aspirated and blocked with
0.25% human serum albumin in PBS buffer for at least 1 hour at room
temperature, then stored desiccated at 4.degree. C. until use. The
plates are rehydrated with wash buffer (Tris-buffered saline, 0.05%
Tween 20) prior to use. The samples and standards are added to the
plates and incubated overnight at 4.degree. C. The plates are
washed three times with wash buffer between each step of the assay.
The biotinylated 3D6, diluted to 0.5 .mu.g/ml in casein incubation
buffer (0.25% casein, PBS, 0.05% Tween 20, pH 7.4) is incubated in
the well for 1 hour at room temperature. Avidin-HRP (Vector,
Burlingame Calif.) diluted 1:4000 in casein incubation buffer is
added to the wells for 1 hour at room temperature. The colorimetric
substrate, Slow TMB-ELISA (Pierce, Cambridge, Mass.), is added and
allowed to react for 15 minutes, after which the enzymatic reaction
is stopped with addition of 2 N H.sub.2SO.sub.4. Reaction product
is quantified using a Molecular Devices Vmax (Molecular Devices,
Santa Clara, Calif.) measuring the difference in absorbance at 450
nm and 650 nm.
2. Blood Assay
[0546] The EDTA plasma is diluted 1:1 in specimen diluent (0.2 gm/l
sodium phosphate. H.sub.2O (monobasic), 2.16 gm/l sodium phosphate.
7H.sub.2O (dibasic), 0.5 gm/l thimerosal, 8.5 gm/l sodium chloride,
0.5 ml Triton X-405, 6.0 g/l globulin-free bovine serum albumin;
and water). The samples and standards in specimen diluent are
assayed using the total .beta.-amyloid assay (266 capture/3D6
reporter) described above for the brain assay except the specimen
diluent is used instead of the casein diluents described.
[0547] Although the invention has been described with reference to
the presently preferred embodiments, it should be understood that
various modifications can be made without departing from the scope
of the invention.
* * * * *