U.S. patent application number 15/104821 was filed with the patent office on 2016-11-03 for 3-(5-chloro-2-oxobenzo[d]oxazol-3(2h)-yl) propanoic acid derivatives as kmo inhibitors.
This patent application is currently assigned to GLAXOSMITHKLINE INTELLECTUAL PROPERTY DEVELOPMENT LIMITED. The applicant listed for this patent is GLAXOSMITHKLINE INTELLECTUAL PROPERTY DEVELOPMENT LIMITED. Invention is credited to Anne Marie Jeanne BOUILLOT, John LIDDLE, Olivier MIRGUET, Anne Louise WALKER.
Application Number | 20160318884 15/104821 |
Document ID | / |
Family ID | 50071094 |
Filed Date | 2016-11-03 |
United States Patent
Application |
20160318884 |
Kind Code |
A1 |
BOUILLOT; Anne Marie Jeanne ;
et al. |
November 3, 2016 |
3-(5-CHLORO-2-OXOBENZO[D]OXAZOL-3(2H)-YL) PROPANOIC ACID
DERIVATIVES AS KMO INHIBITORS
Abstract
A compound of formula (I) or a salt thereof are provided:
##STR00001## wherein R.sup.1, X and R.sup.3 are defined in the
specification, useful in the treatment of disorders mediated by KMO
such as acute pancreatitis, chronic kidney disease, other
conditions associated with systemic inflammatory response syndrome
(SIRS), Huntington's disease, Alzheimer's disease, spinocerebellar
ataxias, Parkinson's disease, AIDS-dementia complex, amylotrophic
lateral sclerosis (ALS), depression, schizophrenia, sepsis,
cardiovascular shock, severe trauma, acute lung injury, acute
respiratory distress syndrome, acute cholecystitis, severe burns,
pneumonia, extensive surgical procedures, ischemic bowel, severe
acute hepatic disease, severe acute hepatic encephalopathy or acute
renal failure.
Inventors: |
BOUILLOT; Anne Marie Jeanne;
(Les Ulis, FR) ; MIRGUET; Olivier; (Les Ulis,
FR) ; LIDDLE; John; (Stevenage, Hertfordshire,
GB) ; WALKER; Anne Louise; (Stevenage, Hertfordshire,
GB) |
|
Applicant: |
Name |
City |
State |
Country |
Type |
GLAXOSMITHKLINE INTELLECTUAL PROPERTY DEVELOPMENT LIMITED |
Brentford, Middlesex |
|
GB |
|
|
Assignee: |
GLAXOSMITHKLINE INTELLECTUAL
PROPERTY DEVELOPMENT LIMITED
Brentford, Middlesex
GB
|
Family ID: |
50071094 |
Appl. No.: |
15/104821 |
Filed: |
December 17, 2014 |
PCT Filed: |
December 17, 2014 |
PCT NO: |
PCT/EP2014/078221 |
371 Date: |
June 15, 2016 |
Current U.S.
Class: |
1/1 |
Current CPC
Class: |
C07C 223/02 20130101;
A61P 25/18 20180101; A61P 13/12 20180101; A61P 31/04 20180101; C07C
309/30 20130101; A61P 1/18 20180101; A61P 31/12 20180101; A61P 9/00
20180101; A61P 25/28 20180101; A61P 43/00 20180101; A61P 21/00
20180101; C07C 215/10 20130101; C07C 211/13 20130101; C07C 279/14
20130101; A61P 25/24 20180101; A61P 11/00 20180101; A61P 1/04
20180101; C07C 211/27 20130101; A61P 25/00 20180101; A61P 1/00
20180101; A61P 17/02 20180101; A61P 25/14 20180101; A61P 25/16
20180101; C07D 413/12 20130101; A61P 1/16 20180101; A61P 37/02
20180101; C07C 229/26 20130101; C07D 263/58 20130101 |
International
Class: |
C07D 263/58 20060101
C07D263/58; C07C 215/10 20060101 C07C215/10; C07C 211/27 20060101
C07C211/27; C07C 211/13 20060101 C07C211/13; C07C 279/14 20060101
C07C279/14; C07C 229/26 20060101 C07C229/26; C07C 309/30 20060101
C07C309/30; C07D 413/12 20060101 C07D413/12; C07C 223/02 20060101
C07C223/02 |
Foreign Application Data
Date |
Code |
Application Number |
Dec 19, 2013 |
GB |
1322512.3 |
Claims
1. (canceled)
2. A compound of formula (I) or a salt thereof ##STR00104##
wherein: X is a bond and R.sup.1 is --H, -halo or --CN; or X is
--CH.sub.2-- and R.sup.1 is --H or --C.sub.1-3alkyl; or X is --O--
and R.sup.1 is --C.sub.1-4alkyl, --(CH.sub.2).sub.mCF.sub.3,
--CHR.sup.2CH.sub.2OMe, --(CH.sub.2).sub.nC.sub.3-4cycloalkyl,
--(CH.sub.2).sub.noxetane, -benzyl or --CHR.sup.2heteroaryl;
wherein heteroaryl may be additionally substituted by halo, methyl,
ethyl or O; m=1 or 2; n=0 or 1; R.sup.2=--H, -methyl or -ethyl; and
R.sup.3.dbd.H or -methyl, provided that the compound of formula (I)
or a salt thereof is not
3-(5-chloro-2-oxobenzo[d]oxazol-3(2H)-yl)propanoic acid or
3-(5-chloro-2-oxobenzo[d]oxazol-3(2H)-yl)-2-methylpropanoic
acid.
3. A compound of formula (I) as defined in claim 2 wherein X is
--CH.sub.2-- and R.sup.1 is --H or --C.sub.1-3alkyl.
4. A compound of formula (I) as defined in claim 2 wherein X is
--CH.sub.2-- and R.sup.1 is --H.
5. A compound of formula (I) as defined in claim 2 wherein X is
--O-- and R.sup.1 is --C.sub.1-4alkyl, --(CH.sub.2).sub.mCF.sub.3,
--CHR.sup.2CH.sub.2OMe, --(CH.sub.2).sub.nC.sub.3-4cycloalkyl,
--(CH.sub.2).sub.noxetane, -benzyl or --CHR.sup.2heteroaryl;
wherein heteroaryl may be additionally substituted by methyl or
O.
6. A compound of formula (I) as defined in claim 2, wherein X is
--O-- and R1 is --CHR2heteroaryl.
7. A compound of formula (I) or a salt thereof which is a compound
selected from the group consisting of:
3-(5-chloro-2-oxobenzo[d]oxazol-3(2H)-yl)propanoic acid;
3-(5-chloro-6-methyl-2-oxobenzo[d]oxazol-3(2H)-yl)propanoic acid;
3-(5-chloro-2-oxo-6-(1-(pyridin-2-yl)ethoxy)benzo[d]oxazol-3(2H)-yl)propa-
noic acid;
(R)-3-(5-chloro-2-oxo-6-(1-(pyridin-2-yl)ethoxy)benzo[d]oxazol--
3(2H)-yl)propanoic acid;
(S)-3-(5-chloro-2-oxo-6-(1-(pyridin-2-yl)ethoxy)benzo[d]oxazol-3(2H)-yl)p-
ropanoic acid;
3-(5,6-dichloro-2-oxobenzo[d]oxazol-3(2H)-yl)propanoic acid;
3-(5-chloro-6-fluoro-2-oxobenzo[d]oxazol-3(2H)-yl)propanoic acid;
3-(5-chloro-6-methoxy-2-oxobenzo[d]oxazol-3(2H)-yl)propanoic acid;
3-(5-chloro-6-cyano-2-oxobenzo[d]oxazol-3(2H)-yl)propanoic acid;
3-(6-bromo-5-chloro-2-oxobenzo[d]oxazol-3(2H)-yl)propanoic acid;
3-(5-chloro-6-ethoxy-2-oxobenzo[d]oxazol-3(2H)-yl)propanoic acid;
3-(5-chloro-6-ethyl-2-oxobenzo[d]oxazol-3(2H)-yl)propanoic acid;
3-(5-chloro-6-isobutoxy-2-oxobenzo[d]oxazol-3(2H)-yl)propanoic
acid;
3-(5-chloro-6-(cyclopropylmethoxy)-2-oxobenzo[d]oxazol-3(2H)-yl)propanoic
acid;
3-(5-chloro-6-(cyclobutylmethoxy)-2-oxobenzo[d]oxazol-3(2H)-yl)prop-
anoic acid;
3-(6-(benzyloxy)-5-chloro-2-oxobenzo[d]oxazol-3(2H)-yl)propanoic
acid;
3-(5-chloro-6-(2-methoxyethoxy)-2-oxobenzo[d]oxazol-3(2H)-yl)propanoic
acid; 3-(5-chloro-2-oxo-6-propoxybenzo[d]oxazol-3(2H)-yl)propanoic
acid;
3-(5-chloro-2-oxo-6-(3,3,3-trifluoropropoxy)benzo[d]oxazol-3(2H)-yl)propa-
noic acid;
3-(5-chloro-2-oxo-6-(pyridin-2-ylmethoxy)benzo[d]oxazol-3(2H)-y-
l)propanoic acid;
3-(5-chloro-6-isopropoxy-2-oxobenzo[d]oxazol-3(2H)-yl)propanoic
acid;
3-(5-chloro-6-cyclobutoxy-2-oxobenzo[d]oxazol-3(2H)-yl)propanoic
acid;
3-(5-chloro-6-cyclopropoxy-2-oxobenzo[d]oxazol-3(2H)-yl)propanoic
acid;
3-(5-chloro-2-oxo-6-(2,2,2-trifluoroethoxy)benzo[d]oxazol-3(2H)-yl)propan-
oic acid;
3-(5-chloro-6-(oxetan-3-yloxy)-2-oxobenzo[d]oxazol-3(2H)-yl)prop-
anoic acid;
3-(5-chloro-6-isobutyl-2-oxobenzo[d]oxazol-3(2H)-yl)propanoic acid;
3-(5-chloro-6-(oxetan-3-ylmethoxy)-2-oxobenzo[d]oxazol-3(2H)-yl)pro-
panoic acid;
3-(5-chloro-6((1-methoxypropan-2-yl)oxy)-2-oxobenzo[d]oxazol-3(2H)-yl)pro-
panoic acid; 3-(5-chloro-6-((6-methyl
pyridin-3-yl)methoxy)-2-oxobenzo[d]oxazol-3(2H)-yl)propanoic acid;
3-(5-chloro-6-((2-methyl
pyridin-4-yl)methoxy)-2-oxobenzo[d]oxazol-3(2H)-yl)propanoic acid;
2-(((3-(2-carboxyethyl)-5-chloro-2-oxo-2,3-dihydrobenzo[d]oxazol-6-yl)oxy-
)methyl)pyridine 1-oxide;
3-(5-chloro-6((1-methyl-1H-imidazol-2-yl)methoxy)-2-oxobenzo[d]oxazol-3(2-
H)-yl)propanoic acid; 3-(5-chloro-6-((2-methyl
pyridin-3-yl)methoxy)-2-oxobenzo[d]oxazol-3(2H)-yl)propanoic acid;
3-(5-chloro-2-oxo-6-(pyrimidin-2-ylmethoxy)benzo[d]oxazol-3(2H)-yl)propan-
oic acid;
3-(5-chloro-6((2-methyloxazol-5-yl)methoxy)-2-oxobenzo[d]oxazol--
3(2H)-yl)propanoic acid;
3-(5-chloro-6((5-methyloxazol-2-yl)methoxy)-2-oxobenzo[d]oxazol-3(2H)-yl)-
propanoic acid;
3-(6-((1H-imidazol-2-yl)methoxy)-5-chloro-2-oxobenzo[d]oxazol-3(2H)-yl)pr-
opanoic acid; and
3-(5-chloro-2-oxo-6-(pyridazin-3-ylmethoxy)benzo[d]oxazol-3(2H)-yl)propan-
oic acid;
(R)-3-(5-chloro-2-oxo-6-(1-(pyridin-2-yl)ethoxy)benzo[d]oxazol-3-
(2H)-yl)propanoic acid;
3-(5-chloro-6-(1-(2-methyloxazol-5-yl)ethoxy)-2-oxobenzo[d]oxazol-3(2H)-y-
l)propanoic acid (racemic);
(R)-3-(5-chloro-6-(1-(5-fluoropyridin-2-yl)ethoxy)-2-oxobenzo[d]oxazol-3(-
2H)-yl)propanoic acid;
(R)-3-(5-chloro-6-(1-(5-methylpyridin-2-yl)ethoxy)-2-oxobenzo[d]oxazol-3(-
2H)-yl)propanoic acid;
3-(5-chloro-6((5-fluoropyridin-2-yl)methoxy)-2-oxobenzo[d]oxazol-3(2H)-yl-
)propanoic acid;
3-(5-chloro-6((5-chloropyridin-2-yl)methoxy)-2-oxobenzo[d]oxazol-3(2H)-yl-
)propanoic acid;
(R)-3-(5-chloro-6-(1-(5-chloropyridin-2-yl)ethoxy)-2-oxobenzo[d]oxazol-3(-
2H)-yl)propanoic acid;
3-(5-chloro-6((5-methylpyridin-2-yl)methoxy)-2-oxobenzo[d]oxazol-3(2H)-yl-
)propanoic acid;
(R)-3-(5-chloro-6-(1-(6-methylpyridin-2-yl)ethoxy)-2-oxobenzo[d]oxazol-3(-
2H)-yl)propanoic acid;
(R)-3-(5-chloro-2-oxo-6-(1-(pyridin-2-yl)propoxy)benzo[d]oxazol-3(2H)-yl)-
propanoic acid;
(R)-3-(5-chloro-6-(1-(4-methylpyridin-2-yl)ethoxy)-2-oxobenzo[d]oxazol-3(-
2H)-yl)propanoic acid;
(R)-3-(5-chloro-2-oxo-6-(1-(pyridazin-3-yl)ethoxy)benzo[d]oxazol-3(2H)-yl-
)propanoic acid;
(S)-3-(5-chloro-2-oxo-6-(1-(pyridazin-3-yl)ethoxy)benzo[d]oxazol-3(2H)-yl-
)propanoic acid;
3-(5-chloro-6((6-methylpyridazin-3-yl)methoxy)-2-oxobenzo[d]oxazol-3(2H)--
yl)propanoic acid;
(S)-3-(5-chloro-6-(1-(6-methylpyridazin-3-yl)ethoxy)-2-oxobenzo[d]oxazol--
3(2H)-yl)propanoic acid;
(R)-3-(5-chloro-6-(1-(6-methylpyridazin-3-yl)ethoxy)-2-oxobenzo[d]oxazol--
3(2H)-yl)propanoic acid;
(R)-3-(5-chloro-6-(1-(5-methylpyridin-2-yl)propoxy)-2-oxobenzo[d]oxazol-3-
(2H)-yl)propanoic acid;
(S)-3-(5-chloro-6-(1-(5-methylpyridin-2-yl)ethoxy)-2-oxobenzo[d]oxazol-3(-
2H)-yl)-2-methylpropanoic acid;
(R)-3-(5-chloro-6-(1-(5-methylpyridin-2-yl)ethoxy)-2-oxobenzo[d]oxazol-3(-
2H)-yl)-2-methylpropanoic acid;
(S)-3-(5-chloro-6-(1-(5-methylpyridin-2-yl)ethoxy)-2-oxobenzo[d]oxazol-3(-
2H)-yl)-2-methylpropanoic acid;
(R)-3-(5-chloro-6-(1-(5-methylpyridin-2-yl)ethoxy)-2-oxobenzo[d]oxazol-3(-
2H)-yl)-2-methylpropanoic acid;
3-(5-chloro-2-oxo-6-(1-(pyrimidin-2-yl)ethoxy)benzo[d]oxazol-3(2H)-yl)pro-
panoic acid (single unknown enantiomer);
(S)-3-(5-chloro-6-(1-(oxazol-2-yl)ethoxy)-2-oxobenzo[d]oxazol-3(2H)-yl)pr-
opanoic acid; and
(R)-3-(5-chloro-6-(1-(oxazol-2-yl)ethoxy)-2-oxobenzo[d]oxazol-3(2H)-yl)pr-
opanoic acid or a pharmaceutically acceptable salt thereof.
8. A compound of formula (I) which is
3-(5-chloro-6-methyl-2-oxobenzo[d]oxazol-3(2H)-yl)propanoic acid of
formula ##STR00105## or a pharmaceutically acceptable salt
thereof.
9. A compound of formula (I) or a pharmaceutically acceptable salt
thereof which is the 2-amino-2-(hydroxymethyl)-1,3-propanediol salt
of 3-(5-chloro-6-methyl-2-oxobenzo[d]oxazol-3(2H)-yl)propanoic acid
of formula ##STR00106##
10. (canceled)
11. (canceled)
12. (canceled)
13. (canceled)
14. (canceled)
15. A method of treatment of conditions or disorders mediated by
KMO which comprises administering to a host in need thereof an
effective amount of a compound of formula (I) or a pharmaceutically
acceptable salt thereof as defined in claim 1 or claim 7.
16. A method of treatment of acute pancreatitis, other conditions
associated with systemic inflammatory response syndrome (SIRS),
Huntington's disease, Alzheimer's disease, spinocerebellar ataxias,
Parkinson's disease, AIDS-dementia complex, amylotrophic lateral
sclerosis (ALS), depression, schizophrenia, sepsis, cardiovascular
shock, severe trauma, acute lung injury, acute respiratory distress
syndrome, acute cholecystitis, severe burns, pneumonia, extensive
surgical procedures, ischemic bowel, severe acute hepatic disease,
severe acute hepatic encephalopathy, or acute renal failure which
comprises administering to a host in need thereof an effective
amount of a compound of formula (I) or a pharmaceutically
acceptable salt thereof as defined in claim 1 or claim 7.
17. A pharmaceutical composition comprising a) a compound as
defined in claim 1 or a pharmaceutically acceptable salt thereof,
and b) one or more pharmaceutically acceptable excipients.
18. A pharmaceutical composition according to claim 17 comprising
3-(5-chloro-6-methyl-2-oxobenzo[d]oxazol-3(2H)-yl)propanoic acid or
a pharmaceutically acceptable salt thereof.
19. A method of treating acute pancreatitis, other conditions
associated with systemic inflammatory response syndrome (SIRS),
Huntington's disease, Alzheimer's disease, spinocerebellar ataxias,
Parkinson's disease, AIDS-dementia complex, amylotrophic lateral
sclerosis (ALS), depression, schizophrenia, sepsis, cardiovascular
shock, severe trauma, acute lung injury, acute respiratory distress
syndrome, acute cholecystitis, severe burns, pneumonia, extensive
surgical procedures, ischemic bowel, severe acute hepatic disease,
severe acute hepatic encephalopathy or acute renal failure in a
host, the method comprising administering to the host a
pharmaceutical composition which comprises a compound of formula
(I) or a pharmaceutically acceptable salt thereof as defined in
claim 1 or claim 7 and one or more pharmaceutically acceptable
excipients.
Description
FIELD OF THE INVENTION
[0001] The present invention relates to novel
5-chlorobenzo[d]oxazol-2(3H)-one derivatives having pharmacological
activity, processes for their preparation, pharmaceutical
compositions containing them and their use in the treatment of
various disorders.
BACKGROUND OF THE INVENTION
[0002] Kynurenine monooxygenase (KMO) is a flavin adenine
dinucleotide (FAD) dependent monooxygenase located on the outer
mitochondrial membrane. KMO is known to oxidise L-Kynurenine (KYN)
to 3-hydroxykynurenine (3HK) as part of the major route of
catabolism of tryptophan. 3HK is then converted to
3-hydroxyanthranilic acid and quinolinic acid by kynureninase
(KYNU) and 3-hydroxyanthranilate 3,4-dioxygenase (3-HAAO).
[0003] KMO is highly expressed in tissues including the liver,
placenta, kidney [Alberati-Giani, FEBS Lett. 410:407-412(1997)]
endothelial cells and monocytes and at a lower level in microglia
and macrophages in the brain.
[0004] Increased levels of 3HK and quinolinic acid and reduced
levels of Kynurenic acid (KYNA), which is formed from kynurenine by
an alternative pathway, have been implicated in a number of
diseases including Huntington's Disease, Parkinson's Disease,
Alzheimer's Disease, amyotrophic lateral sclerosis (ALS) [Amaral,
Outeiro et Al. Journal of Molecular medicine 2013: 91(6): 705-713]
and Acute Pancreatitis [Mole, McFerran et al. British Journal of
Surgery 2008: 95: 855-867]. In the CNS 3-HK and quinolinic acid
have been shown to be neurotoxic and KYNA to have neuroprotective
effects. Inhibition of KMO oxidative activity would therefore be
expected to result in reduced levels of 3-HK and quinolinic acid
and increased levels of KYNA and to potentially show benefit for
these diseases.
[0005] There is a large body of evidence showing that tryptophan
metabolism is also altered in a range of acute injury settings. For
instance, increased kynurenine levels have been associated with the
development of sepsis following trauma [Pellegrin, 2005, Logters,
2009], while increased levels of both kynurenine and 3-HK correlate
with the development of organ failure in acute pancreatitis [Mole,
McFerran et al. British Journal of Surgery 2008: 95: 855-867]. This
dysregulation of tryptophan metabolism is in part accounted for by
the induction of indolamine 2,3 dioxygenase (IDO, the enzyme that
converts tryptophan to N-formyl-kynurenine)) as part of the
inflammatory cascade, but the development of organ dysfunction
appears dependent on the downstream metabolites [Mole, McFerran et
al. British Journal of Surgery 2008: 95: 855-867]
[0006] Acute pancreatitis (AP) results from local injury to the
organ driven by factors such as excessive alcohol consumption or
gallstones. The arising abdominal pain is extremely severe, and
patients will invariably present to an emergency department rapidly
following onset of an attack, with elevation of serum amylase used
as a diagnostic. In the majority of cases, the disease is self
limiting, and the pain resolves within 24-36 hours. However for the
remaining 20-30% of patients a systemic inflammatory response
occurs, resulting in rapid progression to multiple organ
dysfunction (MOD). This leads to a prolonged stay in ICU (averaging
17 days), with a mortality rate of over 30%. Despite this high
unmet need and the seriousness of the disease, there are no
effective treatments available, with current standard of care being
purely supportive.
[0007] SU1143745 discloses methods for the preparation of
derivatives of 3-(3-benzoxazolonyl)propanoic acid. JP54098330
discloses the reaction of benzoxazolepropionic acid derivatives
with thionyl chloride in the manufacture of tricyclic
oxazoloquinoline derivative or thiazoloquinoline derivative
bactericides for agricultural and horticultural use. Onkol et al,
(J. Faculty of Pharmacy of Gazi University (202), 19(1), 15-24)
discloses 2-oxobenzazolin-3-yl)alkanoic acid derivatives and the
testing of these compounds for antinociceptive activity. Kalcheva V
et al (Godishnik na Sofiiskiya Universitet Sy. Kliment Okhridski,
Khimicheski Fakultet (1972), Volume Date 1969-1970, 64, 33-42)
discloses cyanoethylation of benzoxazolethione, benzoxazolone, and
some benzoxazolone derivatives.
[0008] WO2013016488, WO2011091153, WO2010017132, WO2010017179,
WO2010011302, WO2008022286 and WO2008022281 describe inhibitors of
KMO for targeting neurodegenerative disorders or diseases;
EP1475385, EP1424333 describe inhibitors of KMO for targeting
degenerative and inflammatory conditions. There remains a need for
effective KMO inhibitors which are suitable for intravenous
administration for use in the treatment of acute pancreatitis and
other conditions associated with systemic inflammatory response
syndrome (SIRS).
[0009] A structurally novel class of compounds has now been found
which provides inhibitors of KMO which may be useful in the
treatment of acute pancreatitis and acute conditions associated
with systemic inflammatory response syndrome (SIRS).
SUMMARY OF THE INVENTION
[0010] The present invention therefore provides, in a first aspect,
a compound of formula (I) or a pharmaceutically acceptable salt
thereof for use as a medicament:
##STR00002##
[0011] wherein:
[0012] X is a bond and R.sup.1 is --H, -halo or --CN; or
[0013] X is --CH.sub.2-- and R.sup.1 is --H or --C.sub.1-3alkyl;
or
[0014] X is --O-- and R.sup.1 is --C.sub.1-4alkyl,
--(CH.sub.2).sub.mCF.sub.3, --CHR.sup.2CH.sub.2OMe,
--(CH.sub.2).sub.nC.sub.3-4cycloalkyl, --(CH.sub.2).sub.noxetane,
-benzyl or --CHR.sup.2heteroaryl; wherein heteroaryl may be
additionally substituted by halo, methyl, ethyl or O;
[0015] m=1 or 2;
[0016] n=0 or 1;
[0017] R.sup.2=--H, -methyl or -ethyl; and
[0018] R.sup.3.dbd.H or methyl.
[0019] The term "alkyl" as used herein refers to a straight or
branched alkyl group in all isomeric forms. The term
"C.sub.1-4alkyl" refers to an alkyl group, containing at least 1,
and at most 4 carbon atoms. Examples of such C.sub.1-4alkyl groups
include methyl, ethyl, propyl, iso-propyl, n-butyl, iso-butyl,
sec-butyl, and tert-butyl.
[0020] The term "halogen" as used herein refers to e.g. fluorine
(F), chlorine (Cl), bromine (Br), or iodine (I) and the term "halo"
refers to e.g. fluoro (--F), chloro (--Cl), bromo (--Br) or iodo
(--I).
[0021] The term "heteroaryl" as used herein refers to a 5- or
6-membered unsaturated ring which comprises one or more
heteroatoms. When the term "heteroaryl" represents a 5-membered
group it contains a heteroatom selected from O, N or S and may
optionally contain a further 1 to 3 nitrogen atoms. When the term
"heteroaryl" represents a 6-membered group it contains from 1 to 3
nitrogen atoms. Examples of such 5- or 6-membered heteroaryl rings
include, but are not limited to, pyrrolyl, triazolyl, thiadiazolyl,
tetrazolyl, imidazolyl, pyrazolyl, isothiazolyl, thiazolyl,
isoxazolyl, oxazolyl, oxadiazolyl, furazanyl, furanyl, thienyl,
pyridyl, pyrimidinyl, pyrazinyl, pyridazinyl and triazinyl.
[0022] The term "cycloalkyl" as used herein refers to a carbocyclic
ring comprising carbon atoms. The term "C.sub.3-4cycloalkyl" refers
to a carbocyclic ring containing 3 or 4 carbon atoms, for example
cyclopropyl or cyclobutyl.
[0023] In further aspects of the invention, the invention provides
a compound of formula (I) or a salt thereof, and a pharmaceutical
composition comprising the compound of formula (I) or a
pharmaceutically acceptable salt thereof and a pharmaceutically
acceptable carrier.
DETAILED DESCRIPTION OF THE INVENTION
[0024] As discussed above, in a first aspect, the invention
provides a compound of formula (I) or a pharmaceutically acceptable
salt thereof for use as a medicament.
##STR00003##
[0025] wherein:
[0026] X is a bond and R.sup.1 is --H, -halo or --CN; or
[0027] X is --CH.sub.2-- and R.sup.1 is --H or --C.sub.1-3alkyl;
or
[0028] X is --O-- and R.sup.1 is --C.sub.1-4alkyl,
--(CH.sub.2).sub.mCF.sub.3, --CHR.sup.2CH.sub.2OMe,
--(CH.sub.2).sub.nC.sub.3-4cycloalkyl, --(CH.sub.2).sub.noxetane,
-benzyl or --CHR.sup.2heteroaryl; wherein heteroaryl may be
additionally substituted by halo, methyl, ethyl or O;
[0029] m=1 or 2;
[0030] n=0 or 1;
[0031] R.sup.2=--H, -methyl or -ethyl; and
[0032] R.sup.3.dbd.H or methyl.
[0033] In a further aspect, the invention provides a compound of
formula (I') or a pharmaceutically acceptable salt thereof for use
as a medicament.
##STR00004##
[0034] wherein:
[0035] X is a bond and R.sup.1 is --H, -halo or --CN; or
[0036] X is --CH.sub.2-- and R.sup.1 is --H or --C.sub.1-3alkyl;
or
[0037] X is --O-- and R.sup.1 is --C.sub.1-4alkyl,
--(CH.sub.2).sub.mCF.sub.3, --CHR.sup.2CH.sub.2OMe,
--(CH.sub.2).sub.nC.sub.3-4cycloalkyl, --(CH.sub.2).sub.noxetane,
-benzyl or --CHR.sup.2heteroaryl; wherein heteroaryl may be
additionally substituted by methyl or O;
[0038] m=1 or 2;
[0039] n=0 or 1; and
[0040] R.sup.2=--H or -methyl.
[0041] Compounds of formula (I) or pharmaceutically acceptable
salts thereof are inhibitors of KMO activity and are thus believed
to be of potential use in the treatment of conditions or disorders
mediated by KMO.
[0042] Such conditions or disorders include acute pancreatitis,
other conditions associated with systemic inflammatory response
syndrome (SIRS), Huntington's disease, Alzheimer's disease,
spinocerebellar ataxias, Parkinson's disease, AIDS-dementia
complex, amylotrophic lateral sclerosis (ALS), depression,
schizophrenia, sepsis, cardiovascular shock, severe trauma, acute
lung injury, acute respiratory distress syndrome, acute
cholecystitis, severe burns, pneumonia, extensive surgical
procedures, ischemic bowel, severe acute hepatic disease, severe
acute hepatic encephalopathy or acute renal failure.
[0043] Additional conditions or disorders include
hyperproliferative diseases of benign or malignant behaviour, in
which cells of various tissues and organs exhibit aberrant patterns
of growth, proliferation, migration, signalling, senescence, and
death. Generally hyperproliferative disease refers to diseases and
disorders associated with the uncontrolled proliferation of cells,
including but not limited to uncontrolled growth of organ and
tissue cells resulting in cancers and benign tumours.
Hyperproliferative disorders associated with endothelial cells can
result in diseases of angiogenesis such as angiomas, endometriosis,
obesity, age-related macular degeneration and various
retinopathies, as well as the proliferation of ECs and smooth
muscle cells that cause restenosis as a consequence of stenting in
the treatment of atherosclerosis. Hyperproliferative disorders
involving fibroblasts (i.e. fibrogenesis) include but are not
limited to disorders of excessive scaring (i.e. fibrosis) such as
age-related macular degeneration, cardiac remodelling and failure
associated with myocardial infarction, excessive wound healing such
as commonly occurs as a consequence of surgery or injury, keloids,
and fibroid tumours and stenting.
[0044] Further such conditions or disorders include transplant
rejection (suppression of T-cells) and graft vs host disease,
chronic kidney disease, systemic inflammatory disorders, brain
inflammatory disorders including malaria and African
trypanosomiasis, stroke, and pneumococcal meningitis.
[0045] Further such indications or disorders include cirrhosis,
chronic pancreatitis, liver fibrosis, lung fibrosis and
ischemia-reperfusion injury
[0046] Further such conditions or disorders further include, for
example, neurodegenerative diseases, psychiatric or neurological
diseases or disorders, Creutzfeld-Jacob disease, trauma-induced
neurodegeneration, high-pressure neurological syndrome, dystonia,
olivopontocerebellar atrophy, multiple sclerosis, epilepsy,
consequences of stroke, cerebral ischemia, ischemic disorders
including stroke (focal ischemia), hypoxia, multi-infarct dementia,
consequences of cerebral trauma or damage, damage to the spinal
cord, dementia such as senile dementia, AIDS-induced
encephalopathy, other infection related encephalopathy, viral or
bacterial meningitis, infectious diseases caused by viral,
bacterial and other parasites, (for example, general central
nervous system (CNS) infections such as viral, bacterial or
parasitic infection, for example, poliomyelitis, Lyme disease
(Borrelia burgdorferi infection)) septic shock, and malaria,
cancers, cancers with cerebral localization, hepatic
encephalopathy, systemic lupus, analgesia and opiate withdrawal
symptoms, feeding behaviour, psychiatric disorders, such as
insomnia, depression, schizophrenia, severe deficit in working
memory, severe deficit in long term memory storage, decrease in
cognition, severe deficit in attention, severe deficit in executive
functioning, slowness in information processing, slowness in neural
activity, anxiety, generalized anxiety disorders, panic anxiety,
obsessive compulsive disorders, social phobia, performance anxiety,
post-traumatic stress disorder, acute stress reaction, adjustment
reaction, separation anxiety disorder, alcohol withdrawal anxiety,
depressive disorders, disorders of the developing or aged brain,
diabetes, and complications thereof, Tourette's syndrome, Fragile X
syndrome, autism spectrum disorders, disorders that cause severe
and pervasive impairment in thinking feeling, language and the
ability to relate to others, mood disorders, psychological
disorders characterized by abnormalities of emotional state, such
as without limitation, bipolar disorder, unipolar depression, major
depression, endogenous depression, involutional depression,
reactive depression, psychotic depression, depression caused by
underlying medical conditions, depressive disorders, cyclothymic
disorders, dysthymic disorders, mood disorders due to general
medical condition, mood disorders not otherwise specified and
substance-induced mood disorders.
[0047] Further such conditions or disorders also include, for
example, acute pancreatitis, acute necrotizing pancreatitis, AIDS
(disease), analgesia, aseptic meningitis, brain disease, for
example, Gilles de la Tourette syndrome, Asperger syndrome, Rett
syndrome, pervasive developmental disorders, aging-related brain
disease, and developmental brain disease, burnout syndrome, carbon
monoxide poisoning, cardiac arrest or insufficiency and hemorrhagic
shock (global brain ischemia), cataract formation and aging of the
eye, central nervous system disease, cerebrovascular disease,
chronic fatigue syndrome, chronic stress, cognitive disorders,
convulsive disorders, such as variants of grand mal and petit mal
epilepsy and Partial Complex Epilepsy, diabetes mellitus, disease
of the nervous system (e.g., dyskinesia, L-DOPA induced movement
disorders, drug addiction, pain and cataract), drug dependence,
drug withdrawal, feeding disorders, Guillain Barr Syndrome and
other neuropathies, Hepatic encephalopathy, immune disease,
immunitary disorders and therapeutic treatment aimed at modifying
biological responses (for instance administrations of interferons
or interleukins), inflammatory disorders of the central and/or
peripheral nervous system, Injury (trauma, polytrauma), Mental and
behavioral disorders, metabolic disease, pain disease, or disorder
selected from a group of inflammatory pain, neurophathic pain or
migraine, allodynia, hyperalgesia pain, phantom pain, neuropathic
pain related to diabetic neuropathy, multiple organ failure, near
drowning, necrosis, neoplasms of the brain, neoplastic disorders
including lymphomas and other malignant blood disorders, nervous
system disease (high-pressure neurological Syndrome, infection),
nicotine addiction and other addictive disorders including
alcoholism, cannabis, benzodiazepine, barbiturate, morphine and
cocaine dependence, change in appetite, sleep disorders, changes in
sleep pattern, lack of energy, fatigue, low self-esteem,
self-reproach inappropriate guilt, frequent thoughts of death or
suicide, plans or attempts to commit suicide, feelings of
hopelessness and worthlessness, psychomotor agitation or
retardation, diminished capacity for thinking, concentration, or
decisiveness, as a Neuroprotective agent, pain, posttraumatic
stress disorder, sepsis, spinal cord disease, spinocerebellar
ataxia, systemic lupus erythematosis, traumatic damage to the brain
and spinal cord, and tremor syndromes and different movement
disorders (dyskinesia), poor balance, brakykinesia, rigidity,
tremor, change in speech, loss of facial expression, micrographia,
difficulty swallowing, drooling, dementia, confusion, fear, sexual
dysfunction, language impairment, impairment in decision making,
violent outbursts, aggression, hallucination, apathy, impairment in
abstract thinking.
[0048] Further such conditions or disorders also include, for
example, cardiovascular diseases, which refer to diseases and
disorders of the heart and circulatory system. These diseases are
often associated with dyslipoproteinemias and/or dyslipidemias.
Cardiovascular diseases include, but are not limited to,
cardiomegaly, atherosclerosis, myocardial infarction, and
congestive heart failure, coronary heart disease, hypertension and
hypotension.
[0049] In particular, such conditions or disorders include
conditions or disorders where elevated levels of tryptophan
metabolites have been correlated with severity of disease and poor
prognosis, including shock, trauma in patients with multiple organ
failure, severe acute pancreatitis and chronic kidney disease
(Logters, T. T., et al. (2009) Shock 32: 29-34, Dabrowski et al
(2014) Inflammation 37: 223-234, Changsirivathanathamrong et al
(2011) Critical Care Medicine 39: 2678-2683, Mole, D. J., et al.
(2008) Br J Surg 95: 855-867, Zhao (2013) Renal Failure 35:
648-653, Pawlak, K. et al (2009) Blood Coagulation and Fibrinolysis
20: 590-594, Kabayashi, T. et al (2014) Biochemical and Biophysical
Research Communications 445: 412-416).
[0050] Accordingly, in a second aspect, the invention provides a
compound of formula (I) or a pharmaceutically acceptable salt
thereof wherein X and R.sup.1 are as defined above for formula (I)
for use in the treatment of any one of the above conditions or
disorders, and in particular acute pancreatitis.
[0051] The invention further provides a compound of formula (I) or
a pharmaceutically acceptable salt thereof wherein X and R.sup.1
are as defined above for formula (I) for use in the prophylaxis of
any one of the above conditions or disorders, and in particular
acute pancreatitis.
[0052] The invention further provides a method of treatment of the
above conditions or disorders, particularly acute pancreatitis, in
mammals including humans, which comprises administering to the
sufferer a therapeutically effective amount of a compound of
formula (I) or a pharmaceutically acceptable salt thereof wherein X
and R.sup.1 are as defined above for formula (I).
[0053] The invention also provides the use of a compound of formula
(I) or a pharmaceutically acceptable salt thereof wherein X and
R.sup.1 are as defined above for formula (I) in the manufacture of
a medicament for use in the treatment of the above conditions or
disorders and particularly acute pancreatitis.
[0054] In a third aspect, the invention provides a compound of
formula (I) or a salt thereof
##STR00005##
[0055] wherein:
[0056] X is a bond and R.sup.1 is --H, -halo or --CN; or
[0057] X is --CH.sub.2-- and R.sup.1 is --H or --C.sub.1-3alkyl;
or
[0058] X is --O-- and R.sup.1 is --C.sub.1-4alkyl,
--(CH.sub.2).sub.mCF.sub.3, --CHR.sup.2CH.sub.2OMe,
--(CH.sub.2).sub.nC.sub.3-4cycloalkyl, --(CH.sub.2).sub.noxetane,
-benzyl or --CHR.sup.2heteroaryl; wherein heteroaryl may be
additionally substituted by halo, methyl, ethyl or O;
[0059] m=1 or 2;
[0060] n=0 or 1;
[0061] R.sup.2=--H, -methyl or -ethyl; and
[0062] R.sup.3.dbd.H or -methyl,
[0063] provided that the compound of formula (I) or a salt thereof
is not 3-(5-chloro-2-oxobenzo[d]oxazol-3(2H)-yl)propanoic acid or
3-(5-chloro-2-oxobenzo[d]oxazol-3(2H)-yl)-2-methylpropanoic
acid.
[0064] In a further aspect, the invention provides a compound of
formula (IA) or a salt thereof
##STR00006##
[0065] wherein:
[0066] X is a bond and R.sup.1 is -halo or --CN; or
[0067] X is --CH.sub.2-- and R.sup.1 is --H or --C.sub.1-3alkyl;
or
[0068] X is --O-- and R.sup.1 is --C.sub.1-4alkyl,
--(CH.sub.2).sub.mCF.sub.3, --CHR.sup.2CH.sub.2OMe,
--(CH.sub.2).sub.nC.sub.3-4cycloalkyl, --(CH.sub.2).sub.noxetane,
-benzyl or --CHR.sup.2heteroaryl; wherein heteroaryl may be
additionally substituted by methyl or O;
[0069] m=1 or 2;
[0070] n=0 or 1; and
[0071] R.sup.2=--H or -methyl.
[0072] In one embodiment, there is provided a compound of formula
(I) or a salt thereof
##STR00007##
[0073] wherein:
[0074] X is a bond and R.sup.1 is -halo or --CN; or
[0075] X is --CH.sub.2-- and R.sup.1 is --H or --C.sub.1-3alkyl;
or
[0076] X is --O-- and R.sup.1 is --C.sub.1-4alkyl,
--(CH.sub.2).sub.mCF.sub.3, --CHR.sup.2CH.sub.2OMe,
--(CH.sub.2).sub.nC.sub.3-4cycloalkyl, --(CH.sub.2).sub.noxetane,
-benzyl or --CHR.sup.2heteroaryl; wherein heteroaryl may be
additionally substituted by halo, methyl, ethyl or O;
[0077] m=1 or 2;
[0078] n=0 or 1;
[0079] R.sup.2=--H, -methyl or -ethyl; and
[0080] R.sup.3.dbd.H or -methyl.
[0081] In one embodiment of the compound of formula (I), X is a
bond and R.sup.1 is --H, -halo or --CN.
[0082] In a particular embodiment, X is a bond and R.sup.1 is --H,
--Cl, --F, --Br or --CN.
[0083] In an alternative embodiment, X is --CH.sub.2-- and R.sup.1
is --H or --C.sub.1-3alkyl.
[0084] In a particular embodiment, X is --CH.sub.2-- and R.sup.1 is
--H, --CH.sub.3 or --CH(CH.sub.3).sub.2.
[0085] In a more particular embodiment, X is --CH.sub.2-- and
R.sup.1 is --H.
[0086] In an alternative embodiment, X is --O-- and R.sup.1 is
--C.sub.1-4alkyl, --(CH.sub.2).sub.mCF.sub.3,
--CHR.sup.2CH.sub.2OMe, --(CH.sub.2).sub.nC.sub.3-4cycloalkyl,
--(CH.sub.2).sub.noxetane, -benzyl or --CHR.sup.2heteroaryl;
wherein heteroaryl may be additionally substituted by halo, methyl
or O.
[0087] In an alternative embodiment, X is --O-- and R.sup.1 is
--C.sub.1-4alkyl, --(CH.sub.2).sub.mCF.sub.3,
--CHR.sup.2CH.sub.2OMe, --(CH.sub.2).sub.nC.sub.3-4cycloalkyl,
--(CH.sub.2).sub.noxetane, -benzyl or --CHR.sup.2heteroaryl;
wherein heteroaryl may be additionally substituted by methyl or
O.
[0088] In a more particular embodiment, X is --O-- and R.sup.1 is
--CHR.sup.2CH.sub.2OMe, --(CH.sub.2).sub.nC.sub.3-4cycloalkyl, or
--(CH.sub.2).sub.noxetane.
[0089] In a more particular embodiment, X is --O-- and R.sup.1 is
--CHR.sup.2heteroaryl.
[0090] In a more particular embodiment X is --O-- and R.sup.1 is
--CHR.sup.2pyridyl.
[0091] In a more particular embodiment X is --O-- and R.sup.1 is
--CHR.sup.2-pyrid-2-yl.
[0092] In one embodiment, R.sup.2 is H.
[0093] In an alternative embodiment, R.sup.2 is -methyl.
[0094] In an alternative embodiment, R.sup.2 is ethyl.
[0095] In one embodiment R.sup.3 is H. In an alternative embodiment
R.sup.3 is methyl.
[0096] In an alternative embodiment, X is --O-- and R.sup.1 is
1-pyrid-2-ylethyl, methyl, ethyl, isobutyl, cyclopropylmethyl,
cyclobutylmethyl, benzyl, 2-methoxyethyl, propyl,
3,3,3-trifluoropropyl, pyrid-2-ylmethyl, isopropyl, cyclobutyl,
cyclopropyl, 2,2,2-trifluoroethyl, oxetan-3-yl, oxetan-3-ylmethyl,
1-methoxypropan-2-yl, 6-methylpyridin-3-yl,
2-methylpyridin-4-ylmethyl, pyrid-2-ylmethyl-N-oxide,
1-methyl-1H-imidazol-2-ylmethyl, 2-methylpyridin-3-ylmethyl,
pyrimidin-2-ylmethyl, 2-methyloxazol-5-ylmethyl,
5-methyloxazol-2-ylmethyl, 1H-imidazol-2-ylmethyl,
pyridazin-3-ylmethyl, 1-(2-methyloxazol-5-yl)ethyl,
1-(5-fluoropyridin-2-yl)ethyl, 1-(5-methylpyridin-2-yl)ethyl,
5-fluoropropyridin-2-ylmethyl, 5-chloropyridin-2-ylmethyl,
1-(5-chloropyridin-2-yl)ethyl, 5-methylpyridin-2-ylmethyl,
1-(6-methylpyridin-2-yl)ethyl, 1-(pyridine-2-yl)propyl,
1-(4-methylpyridin-2-yl)ethyl, 1-(pyridazin-3-yl)ethyl,
6-methylpyridazin-3-ylmethyl, 1-(6-methylpyridazin-3-yl)ethyl,
1-(5-methylpyridin-2-yl)propyl, 1-(5-methylpyridin-2-yl)ethyl,
1-(4-methylpyridin-2-yl)ethyl, 1-(pyridazin-3-yl)ethyl,
6-methylpyridazin-3-ylmethyl, 1-(6-methylpyridazin-3-yl)ethyl,
1-pyrimidin-2-ylethyl, or 1-oxazol-2-ylethyl.
[0097] In an alternative embodiment, X is --O-- and R.sup.1 is
1-pyrid-2-ylethyl, methyl, ethyl, isobutyl, cyclopropylmethyl,
cyclobutylmethyl, benzyl, 2-methoxyethyl, propyl,
3,3,3-trifluoropropyl, pyrid-2-ylmethyl, isopropyl, cyclobutyl,
cyclopropyl, 2,2,2-trifluoroethyl, oxetan-3-yl, oxetan-3-ylmethyl,
1-methoxypropan-2-yl, 6-methylpyridin-3-yl,
2-methylpyridin-4-ylmethyl, pyrid-2-ylmethyl-N-oxide,
1-methyl-1H-imidazol-2-ylmethyl, 2-methylpyridin-3-ylmethyl,
pyrimidin-2-ylmethyl, 2-methyloxazol-5-ylmethyl,
5-methyloxazol-2-ylmethyl, 1H-imidazol-2-ylmethyl or
pyridazin-3-ylmethyl.
[0098] In an even more particular embodiment, X is --O--, R.sup.1
is --CHR.sup.2-pyrid-2-yl, and R.sup.2 is -methyl.
[0099] In an even more particular embodiment, X is --O--, R.sup.1
is --CHR.sup.2-pyrid-2-yl, R.sup.2 is -methyl and R.sup.3 is H.
[0100] All features and embodiments of compounds of formula (I)
apply to compounds of formula (IA) mutatis mutandis. Hereinafter,
all references to compounds of formula (I) include compounds of
formula (IA).
[0101] All features and embodiments of compounds of formula (I)
apply to compounds of formula (IA), and (I') mutatis mutandis.
Hereinafter, all references to compounds of formula (I) include
compounds of formula (IA), and (I').
[0102] Prodrugs of the compounds of formula (I) are included within
the scope of the present invention. In one embodiment, the
compounds of formula (I) or salts thereof are not prodrugs.
[0103] As used herein, the term "prodrug" means a compound which is
converted within the body, e.g. by hydrolysis in the blood, into
its active form that has medical effects. Pharmaceutically
acceptable prodrugs are described in T. Higuchi and V. Stella,
Prodrugs as Novel Delivery Systems, Vol. 14 of the A.C.S. Symposium
Series, and in Edward B. Roche, ed., Bioreversible Carriers in Drug
Design, American Pharmaceutical Association and Pergamon Press,
1987 and in D. Fleishner, S. Ramon and H. Barba "Improved oral drug
delivery: solubility limitations overcome by the use of prodrugs",
Advanced Drug Delivery Reviews (1996) 19(2) 115-130. Prodrugs are
any covalently bonded carriers that release a compound of formula
(I) in vivo when such prodrug is administered to a patient.
Prodrugs are generally prepared by modifying functional groups in a
way such that the modification is cleaved in vivo yielding the
parent compound. Prodrugs may include, for example, compounds of
this invention wherein the carboxylic acid group is bonded to any
group that, when administered to a patient, cleaves to form the
carboxylic acid group. Thus, representative examples of prodrugs
include (but are not limited to) phosphonate, carbamate, acetate,
formate and benzoate derivatives of the carboxylic acid functional
group of the compounds of formula (I).
[0104] Compounds of formula (I) or salts thereof include the
compounds of Examples 1-61 and their salts.
[0105] In one embodiment, compounds of formula (I) or salts thereof
are selected from the group consisting of: [0106]
3-(5-chloro-2-oxobenzo[d]oxazol-3(2H)-yl)propanoic acid; [0107]
3-(5-chloro-6-methyl-2-oxobenzo[d]oxazol-3(2H)-yl)propanoic acid;
[0108]
3-(5-chloro-2-oxo-6-(1-(pyridin-2-yl)ethoxy)benzo[d]oxazol-3(2H)-yl)propa-
noic acid; [0109]
(R)-3-(5-chloro-2-oxo-6-(1-(pyridin-2-yl)ethoxy)benzo[d]oxazol-3(2H)-yl)p-
ropanoic acid; [0110]
(S)-3-(5-chloro-2-oxo-6-(1-(pyridin-2-yl)ethoxy)benzo[d]oxazol-3(2H)-yl)p-
ropanoic acid; [0111]
3-(5,6-dichloro-2-oxobenzo[d]oxazol-3(2H)-yl)propanoic acid; [0112]
3-(5-chloro-6-fluoro-2-oxobenzo[d]oxazol-3(2H)-yl)propanoic acid;
[0113] 3-(5-chloro-6-methoxy-2-oxobenzo[d]oxazol-3(2H)-yl)propanoic
acid; [0114]
3-(5-chloro-6-cyano-2-oxobenzo[d]oxazol-3(2H)-yl)propanoic acid;
[0115] 3-(6-bromo-5-chloro-2-oxobenzo[d]oxazol-3(2H)-yl)propanoic
acid; [0116]
3-(5-chloro-6-ethoxy-2-oxobenzo[d]oxazol-3(2H)-yl)propanoic acid;
[0117] 3-(5-chloro-6-ethyl-2-oxobenzo[d]oxazol-3(2H)-yl)propanoic
acid; [0118]
3-(5-chloro-6-isobutoxy-2-oxobenzo[d]oxazol-3(2H)-yl)propanoic
acid; [0119]
3-(5-chloro-6-(cyclopropylmethoxy)-2-oxobenzo[d]oxazol-3(2H)-yl)pr-
opanoic acid; [0120]
3-(5-chloro-6-(cyclobutylmethoxy)-2-oxobenzo[d]oxazol-3(2H)-yl)propanoic
acid; [0121]
3-(6-(benzyloxy)-5-chloro-2-oxobenzo[d]oxazol-3(2H)-yl)propanoic
acid; [0122]
3-(5-chloro-6-(2-methoxyethoxy)-2-oxobenzo[d]oxazol-3(2H)-yl)propa-
noic acid; [0123]
3-(5-chloro-2-oxo-6-propoxybenzo[d]oxazol-3(2H)-yl)propanoic acid;
[0124]
3-(5-chloro-2-oxo-6-(3,3,3-trifluoropropoxy)benzo[d]oxazol-3(2H)-yl)propa-
noic acid; [0125]
3-(5-chloro-2-oxo-6-(pyridin-2-ylmethoxy)benzo[d]oxazol-3(2H)-yl)propanoi-
c acid; [0126]
3-(5-chloro-6-isopropoxy-2-oxobenzo[d]oxazol-3(2H)-yl)propanoic
acid; [0127]
3-(5-chloro-6-cyclobutoxy-2-oxobenzo[d]oxazol-3(2H)-yl)propanoic
acid; [0128]
3-(5-chloro-6-cyclopropoxy-2-oxobenzo[d]oxazol-3(2H)-yl)propanoic
acid; [0129]
3-(5-chloro-2-oxo-6-(2,2,2-trifluoroethoxy)benzo[d]oxazol-3(2H)-yl-
)propanoic acid; [0130]
3-(5-chloro-6-(oxetan-3-yloxy)-2-oxobenzo[d]oxazol-3(2H)-yl)propanoic
acid; [0131]
3-(5-chloro-6-isobutyl-2-oxobenzo[d]oxazol-3(2H)-yl)propanoic acid;
[0132]
3-(5-chloro-6-(oxetan-3-ylmethoxy)-2-oxobenzo[d]oxazol-3(2H)-yl)pr-
opanoic acid; [0133]
3-(5-chloro-6-((1-methoxypropan-2-yl)oxy)-2-oxobenzo[d]oxazol-3(2H)-yl)pr-
opanoic acid; [0134]
3-(5-chloro-6-((6-methylpyridin-3-yl)methoxy)-2-oxobenzo[d]oxazol-3(2H)-y-
l)propanoic acid; [0135]
3-(5-chloro-6-((2-methylpyridin-4-yl)methoxy)-2-oxobenzo[d]oxazol-3(2H)-y-
l)propanoic acid; [0136]
2-(((3-(2-carboxyethyl)-5-chloro-2-oxo-2,3-dihydrobenzo[d]oxazol-6-yl)oxy-
)methyl)pyridine 1-oxide; [0137]
3-(5-chloro-6-((1-methyl-1H-imidazol-2-yl)methoxy)-2-oxobenzo[d]oxazol-3(-
2H)-yl)propanoic acid; [0138]
3-(5-chloro-6-((2-methylpyridin-3-yl)methoxy)-2-oxobenzo[d]oxazol-3(2H)-y-
l)propanoic acid; [0139]
3-(5-chloro-2-oxo-6-(pyrimidin-2-ylmethoxy)benzo[d]oxazol-3(2H)-yl)propan-
oic acid; [0140]
3-(5-chloro-6-((2-methyloxazol-5-yl)methoxy)-2-oxobenzo[d]oxazol-3(2H)-yl-
)propanoic acid; [0141]
3-(5-chloro-6-((5-methyloxazol-2-yl)methoxy)-2-oxobenzo[d]oxazol-3(2H)-yl-
)propanoic acid; [0142]
3-(6-((1H-imidazol-2-yl)methoxy)-5-chloro-2-oxobenzo[d]oxazol-3(2H)-yl)pr-
opanoic acid; and [0143]
3-(5-chloro-2-oxo-6-(pyridazin-3-ylmethoxy)benzo[d]oxazol-3(2H)-yl)propan-
oic acid; [0144]
(R)-3-(5-chloro-2-oxo-6-(1-(pyridin-2-yl)ethoxy)benzo[d]oxazol-3(2H)-yl)p-
ropanoic acid; [0145]
3-(5-chloro-6-(1-(2-methyloxazol-5-yl)ethoxy)-2-oxobenzo[d]oxazol-3(2H)-y-
l)propanoic acid (racemic); [0146]
(R)-3-(5-chloro-6-(1-(5-fluoropyridin-2-yl)ethoxy)-2-oxobenzo[d]oxazol-3(-
2H)-yl)propanoic acid; [0147]
(R)-3-(5-chloro-6-(1-(5-methylpyridin-2-yl)ethoxy)-2-oxobenzo[d]oxazol-3(-
2H)-yl)propanoic acid; [0148]
3-(5-chloro-6-((5-fluoropyridin-2-yl)methoxy)-2-oxobenzo[d]oxazol-3(2H)-y-
l)propanoic acid; [0149]
3-(5-chloro-6-((5-chloropyridin-2-yl)methoxy)-2-oxobenzo[d]oxazol-3(2H)-y-
l)propanoic acid; [0150]
(R)-3-(5-chloro-6-(1-(5-chloropyridin-2-yl)ethoxy)-2-oxobenzo[d]oxazol-3(-
2H)-yl)propanoic acid; [0151]
3-(5-chloro-6-((5-methylpyridin-2-yl)methoxy)-2-oxobenzo[d]oxazol-3(2H)-y-
l)propanoic acid; [0152]
(R)-3-(5-chloro-6-(1-(6-methylpyridin-2-yl)ethoxy)-2-oxobenzo[d]oxazol-3(-
2H)-yl)propanoic acid; [0153]
(R)-3-(5-chloro-2-oxo-6-(1-(pyridin-2-yl)propoxy)benzo[d]oxazol-3(2H)-yl)-
propanoic acid; [0154]
(R)-3-(5-chloro-6-(1-(4-methylpyridin-2-yl)ethoxy)-2-oxobenzo[d]oxazol-3(-
2H)-yl)propanoic acid; [0155]
(R)-3-(5-chloro-2-oxo-6-(1-(pyridazin-3-yl)ethoxy)benzo[d]oxazol-3(2H)-yl-
)propanoic acid; [0156]
(S)-3-(5-chloro-2-oxo-6-(1-(pyridazin-3-yl)ethoxy)benzo[d]oxazol-3(2H)-yl-
)propanoic acid; [0157]
3-(5-chloro-6-((6-methylpyridazin-3-yl)methoxy)-2-oxobenzo[d]oxazol-3(2H)-
-yl)propanoic acid; [0158]
(S)-3-(5-chloro-6-(1-(6-methylpyridazin-3-yl)ethoxy)-2-oxobenzo[d]oxazol--
3(2H)-yl)propanoic acid; [0159]
(R)-3-(5-chloro-6-(1-(6-methylpyridazin-3-yl)ethoxy)-2-oxobenzo[d]oxazol--
3(2H)-yl)propanoic acid; [0160]
(R)-3-(5-chloro-6-(1-(5-methylpyridin-2-yl)propoxy)-2-oxobenzo[d]oxazol-3-
(2H)-yl)propanoic acid; [0161]
(S)-3-(5-chloro-6-(1-(5-methylpyridin-2-yl)ethoxy)-2-oxobenzo[d]oxazol-3(-
2H)-yl)-2-methylpropanoic acid; [0162]
(R)-3-(5-chloro-6-(1-(5-methylpyridin-2-yl)ethoxy)-2-oxobenzo[d]oxazol-3(-
2H)-yl)-2-methylpropanoic acid; [0163]
(S)-3-(5-chloro-6-(1-(5-methylpyridin-2-yl)ethoxy)-2-oxobenzo[d]oxazol-3(-
2H)-yl)-2-methylpropanoic acid; [0164]
(R)-3-(5-chloro-6-(1-(5-methylpyridin-2-yl)ethoxy)-2-oxobenzo[d]oxazol-3(-
2H)-yl)-2-methylpropanoic acid; [0165]
3-(5-chloro-2-oxo-6-(1-(pyrimidin-2-yl)ethoxy)benzo[d]oxazol-3(2H)-yl)pro-
panoic acid (single unknown enantiomer); [0166]
(S)-3-(5-chloro-6-(1-(oxazol-2-yl)ethoxy)-2-oxobenzo[d]oxazol-3(2H)-yl)pr-
opanoic acid; and [0167]
(R)-(5-chloro-6-(1-(oxazol-2-yl)ethoxy)-2-oxobenzo[d]oxazol-3(2H)-yl)prop-
anoic acid; [0168] and salts thereof
[0169] In a further embodiment, compounds of formula (I) or salts
thereof are selected from the group consisting of: [0170]
3-(5-chloro-6-methyl-2-oxobenzo[d]oxazol-3(2H)-yl)propanoic acid;
[0171]
3-(5-chloro-2-oxo-6-(1-(pyridin-2-yl)ethoxy)benzo[d]oxazol-3(2H)-yl)propa-
noic acid; [0172]
(R)-3-(5-chloro-2-oxo-6-(1-(pyridin-2-yl)ethoxy)benzo[d]oxazol-3(2H)-yl)p-
ropanoic acid; [0173]
(S)-3-(5-chloro-2-oxo-6-(1-(pyridin-2-yl)ethoxy)benzo[d]oxazol-3(2H)-yl)p-
ropanoic acid; [0174]
3-(5,6-dichloro-2-oxobenzo[d]oxazol-3(2H)-yl)propanoic acid; [0175]
3-(5-chloro-6-fluoro-2-oxobenzo[d]oxazol-3(2H)-yl)propanoic acid;
[0176] 3-(5-chloro-6-methoxy-2-oxobenzo[d]oxazol-3(2H)-yl)propanoic
acid; [0177]
3-(5-chloro-6-cyano-2-oxobenzo[d]oxazol-3(2H)-yl)propanoic acid;
[0178] 3-(6-bromo-5-chloro-2-oxobenzo[d]oxazol-3(2H)-yl)propanoic
acid; [0179]
3-(5-chloro-6-ethoxy-2-oxobenzo[d]oxazol-3(2H)-yl)propanoic acid;
[0180] 3-(5-chloro-6-ethyl-2-oxobenzo[d]oxazol-3(2H)-yl)propanoic
acid; [0181]
3-(5-chloro-6-isobutoxy-2-oxobenzo[d]oxazol-3(2H)-yl)propanoic
acid; [0182]
3-(5-chloro-6-(cyclopropylmethoxy)-2-oxobenzo[d]oxazol-3(2H)-yl)pr-
opanoic acid; [0183]
3-(5-chloro-6-(cyclobutylmethoxy)-2-oxobenzo[d]oxazol-3(2H)-yl)propanoic
acid; [0184]
3-(6-(benzyloxy)-5-chloro-2-oxobenzo[d]oxazol-3(2H)-yl)propanoic
acid; [0185]
3-(5-chloro-6-(2-methoxyethoxy)-2-oxobenzo[d]oxazol-3(2H)-yl)propa-
noic acid; [0186]
3-(5-chloro-2-oxo-6-propoxybenzo[d]oxazol-3(2H)-yl)propanoic acid;
[0187]
3-(5-chloro-2-oxo-6-(3,3,3-trifluoropropoxy)benzo[d]oxazol-3(2H)-yl)propa-
noic acid; [0188]
3-(5-chloro-2-oxo-6-(pyridin-2-ylmethoxy)benzo[d]oxazol-3(2H)-yl)propanoi-
c acid; [0189]
3-(5-chloro-6-isopropoxy-2-oxobenzo[d]oxazol-3(2H)-yl)propanoic
acid; [0190]
3-(5-chloro-6-cyclobutoxy-2-oxobenzo[d]oxazol-3(2H)-yl)propanoic
acid; [0191]
3-(5-chloro-6-cyclopropoxy-2-oxobenzo[d]oxazol-3(2H)-yl)propanoic
acid; [0192]
3-(5-chloro-2-oxo-6-(2,2,2-trifluoroethoxy)benzo[d]oxazol-3(2H)-yl-
)propanoic acid; [0193]
3-(5-chloro-6-(oxetan-3-yloxy)-2-oxobenzo[d]oxazol-3(2H)-yl)propanoic
acid; [0194]
3-(5-chloro-6-isobutyl-2-oxobenzo[d]oxazol-3(2H)-yl)propanoic acid;
[0195]
3-(5-chloro-6-(oxetan-3-ylmethoxy)-2-oxobenzo[d]oxazol-3(2H)-yl)pr-
opanoic acid; [0196]
3-(5-chloro-6-((1-methoxypropan-2-yl)oxy)-2-oxobenzo[d]oxazol-3(2H)-yl)pr-
opanoic acid; [0197]
3-(5-chloro-6-((6-methylpyridin-3-yl)methoxy)-2-oxobenzo[d]oxazol-3(2H)-y-
l)propanoic acid; [0198]
3-(5-chloro-6-((2-methylpyridin-4-yl)methoxy)-2-oxobenzo[d]oxazol-3(2H)-y-
l)propanoic acid; [0199]
2-(((3-(2-carboxyethyl)-5-chloro-2-oxo-2,3-dihydrobenzo[d]oxazol-6-yl)oxy-
)methyl)pyridine 1-oxide; [0200]
3-(5-chloro-6-((1-methyl-1H-imidazol-2-yl)methoxy)-2-oxobenzo[d]oxazol-3(-
2H)-yl)propanoic acid; [0201]
3-(5-chloro-6-((2-methylpyridin-3-yl)methoxy)-2-oxobenzo[d]oxazol-3(2H)-y-
l)propanoic acid; [0202]
3-(5-chloro-2-oxo-6-(pyrimidin-2-ylmethoxy)benzo[d]oxazol-3(2H)-yl)propan-
oic acid; [0203]
3-(5-chloro-6-((2-methyloxazol-5-yl)methoxy)-2-oxobenzo[d]oxazol-3(2H)-yl-
)propanoic acid; [0204]
3-(5-chloro-6-((5-methyloxazol-2-yl)methoxy)-2-oxobenzo[d]oxazol-3(2H)-yl-
)propanoic acid; [0205]
3-(6-((1H-imidazol-2-yl)methoxy)-5-chloro-2-oxobenzo[d]oxazol-3(2H)-yl)pr-
opanoic acid; and [0206]
3-(5-chloro-2-oxo-6-(pyridazin-3-ylmethoxy)benzo[d]oxazol-3(2H)-yl)propan-
oic acid; [0207]
(R)-3-(5-chloro-2-oxo-6-(1-(pyridin-2-yl)ethoxy)benzo[d]oxazol-3(2H)-yl)p-
ropanoic acid; [0208]
3-(5-chloro-6-(1-(2-methyloxazol-5-yl)ethoxy)-2-oxobenzo[d]oxazol-3(2H)-y-
l)propanoic acid (racemic); [0209]
(R)-3-(5-chloro-6-(1-(5-fluoropyridin-2-yl)ethoxy)-2-oxobenzo[d]oxazol-3(-
2H)-yl)propanoic acid; [0210]
(R)-3-(5-chloro-6-(1-(5-methylpyridin-2-yl)ethoxy)-2-oxobenzo[d]oxazol-3(-
2H)-yl)propanoic acid; [0211]
3-(5-chloro-6-((5-fluoropyridin-2-yl)methoxy)-2-oxobenzo[d]oxazol-3(2H)-y-
l)propanoic acid; [0212]
3-(5-chloro-6-((5-chloropyridin-2-yl)methoxy)-2-oxobenzo[d]oxazol-3(2H)-y-
l)propanoic acid; [0213]
(R)-3-(5-chloro-6-(1-(5-chloropyridin-2-yl)ethoxy)-2-oxobenzo[d]oxazol-3(-
2H)-yl)propanoic acid; [0214]
3-(5-chloro-6-((5-methylpyridin-2-yl)methoxy)-2-oxobenzo[d]oxazol-3(2H)-y-
l)propanoic acid; [0215]
(R)-3-(5-chloro-6-(1-(6-methylpyridin-2-yl)ethoxy)-2-oxobenzo[d]oxazol-3(-
2H)-yl)propanoic acid; [0216]
(R)-3-(5-chloro-2-oxo-6-(1-(pyridin-2-yl)propoxy)benzo[d]oxazol-3(2H)-yl)-
propanoic acid; [0217]
(R)-3-(5-chloro-6-(1-(4-methylpyridin-2-yl)ethoxy)-2-oxobenzo[d]oxazol-3(-
2H)-yl)propanoic acid; [0218]
(R)-3-(5-chloro-2-oxo-6-(1-(pyridazin-3-yl)ethoxy)benzo[d]oxazol-3(2H)-yl-
)propanoic acid; [0219]
(S)-3-(5-chloro-2-oxo-6-(1-(pyridazin-3-yl)ethoxy)benzo[d]oxazol-3(2H)-yl-
)propanoic acid; [0220]
3-(5-chloro-6-((6-methylpyridazin-3-yl)methoxy)-2-oxobenzo[d]oxazol-3(2H)-
-yl)propanoic acid; [0221]
(R)-3-(5-chloro-6-(1-(6-methylpyridazin-3-yl)ethoxy)-2-oxobenzo[d]oxazol--
3(2H)-yl)propanoic acid; [0222]
(S)-3-(5-chloro-6-(1-(6-methylpyridazin-3-yl)ethoxy)-2-oxobenzo[d]oxazol--
3(2H)-yl)propanoic acid; [0223]
(R)-3-(5-chloro-6-(1-(5-methylpyridin-2-yl)propoxy)-2-oxobenzo[d]oxazol-3-
(2H)-yl)propanoic acid; [0224]
(R)-3-(5-chloro-6-(1-(5-methylpyridin-2-yl)ethoxy)-2-oxobenzo[d]oxazol-3(-
2H)-yl)-2-methylpropanoic acid; [0225]
(S)-3-(5-chloro-6-(1-(5-methylpyridin-2-yl)ethoxy)-2-oxobenzo[d]oxazol-3(-
2H)-yl)-2-methylpropanoic acid; [0226]
(R)-3-(5-chloro-6-(1-(5-methylpyridin-2-yl)ethoxy)-2-oxobenzo[d]oxazol-3(-
2H)-yl)-2-methylpropanoic acid; [0227]
(S)-3-(5-chloro-6-(1-(5-methylpyridin-2-yl)ethoxy)-2-oxobenzo[d]oxazol-3(-
2H)-yl)-2-methylpropanoic acid; [0228]
(R)-3-(5-chloro-2-oxo-6-(1-(pyrimidin-2-yl)ethoxy)benzo[d]oxazol-3(2H)-yl-
)propanoic acid; [0229]
(S)-3-(5-chloro-6-(1-(oxazol-2-yl)ethoxy)-2-oxobenzo[d]oxazol-3(2H)-yl)pr-
opanoic acid; and [0230]
(R)-3-(5-chloro-6-(1-(oxazol-2-yl)ethoxy)-2-oxobenzo[d]oxazol-3(2H)-yl)pr-
opanoic acid; [0231] and salts thereof
[0232] In one embodiment, compounds of formula (I) or salts thereof
are selected from the group consisting of: [0233]
3-(5-chloro-6-methyl-2-oxobenzo[d]oxazol-3(2H)-yl)propanoic acid;
[0234]
3-(5-chloro-2-oxo-6-(1-(pyridin-2-yl)ethoxy)benzo[d]oxazol-3(2H)-yl)propa-
noic acid; [0235]
(R)-3-(5-chloro-2-oxo-6-(1-(pyridin-2-yl)ethoxy)benzo[d]oxazol-3(2H)-yl)p-
ropanoic acid; [0236]
(S)-3-(5-chloro-2-oxo-6-(1-(pyridin-2-yl)ethoxy)benzo[d]oxazol-3(2H)-yl)p-
ropanoic acid; [0237]
3-(5,6-dichloro-2-oxobenzo[d]oxazol-3(2H)-yl)propanoic acid; [0238]
3-(5-chloro-6-fluoro-2-oxobenzo[d]oxazol-3(2H)-yl)propanoic acid;
[0239] 3-(5-chloro-6-methoxy-2-oxobenzo[d]oxazol-3(2H)-yl)propanoic
acid; [0240]
3-(5-chloro-6-cyano-2-oxobenzo[d]oxazol-3(2H)-yl)propanoic acid;
[0241] 3-(6-bromo-5-chloro-2-oxobenzo[d]oxazol-3(2H)-yl)propanoic
acid; [0242]
3-(5-chloro-6-ethoxy-2-oxobenzo[d]oxazol-3(2H)-yl)propanoic acid;
[0243] 3-(5-chloro-6-ethyl-2-oxobenzo[d]oxazol-3(2H)-yl)propanoic
acid; [0244]
3-(5-chloro-6-isobutoxy-2-oxobenzo[d]oxazol-3(2H)-yl)propanoic
acid; [0245]
3-(5-chloro-6-(cyclopropylmethoxy)-2-oxobenzo[d]oxazol-3(2H)-yl)pr-
opanoic acid; [0246]
3-(5-chloro-6-(cyclobutylmethoxy)-2-oxobenzo[d]oxazol-3(2H)-yl)propanoic
acid; [0247]
3-(6-(benzyloxy)-5-chloro-2-oxobenzo[d]oxazol-3(2H)-yl)propanoic
acid; [0248]
3-(5-chloro-6-(2-methoxyethoxy)-2-oxobenzo[d]oxazol-3(2H)-yl)propa-
noic acid; [0249]
3-(5-chloro-2-oxo-6-propoxybenzo[d]oxazol-3(2H)-yl)propanoic acid;
[0250]
3-(5-chloro-2-oxo-6-(3,3,3-trifluoropropoxy)benzo[d]oxazol-3(2H)-yl)propa-
noic acid; [0251]
3-(5-chloro-2-oxo-6-(pyridin-2-ylmethoxy)benzo[d]oxazol-3(2H)-yl)propanoi-
c acid; [0252]
3-(5-chloro-6-isopropoxy-2-oxobenzo[d]oxazol-3(2H)-yl)propanoic
acid; [0253]
3-(5-chloro-6-cyclobutoxy-2-oxobenzo[d]oxazol-3(2H)-yl)propanoic
acid; [0254]
3-(5-chloro-6-cyclopropoxy-2-oxobenzo[d]oxazol-3(2H)-yl)propanoic
acid; [0255]
3-(5-chloro-2-oxo-6-(2,2,2-trifluoroethoxy)benzo[d]oxazol-3(2H)-yl-
)propanoic acid; [0256]
3-(5-chloro-6-(oxetan-3-yloxy)-2-oxobenzo[d]oxazol-3(2H)-yl)propanoic
acid; [0257]
3-(5-chloro-6-isobutyl-2-oxobenzo[d]oxazol-3(2H)-yl)propanoic acid;
[0258]
3-(5-chloro-6-(oxetan-3-ylmethoxy)-2-oxobenzo[d]oxazol-3(2H)-yl)pr-
opanoic acid; [0259]
3-(5-chloro-6-((1-methoxypropan-2-yl)oxy)-2-oxobenzo[d]oxazol-3(2H)-yl)pr-
opanoic acid; [0260]
3-(5-chloro-6-((6-methylpyridin-3-yl)methoxy)-2-oxobenzo[d]oxazol-3(2H)-y-
l)propanoic acid; [0261]
3-(5-chloro-6-((2-methylpyridin-4-yl)methoxy)-2-oxobenzo[d]oxazol-3(2H)-y-
l)propanoic acid; [0262]
2-(((3-(2-carboxyethyl)-5-chloro-2-oxo-2,3-dihydrobenzo[d]oxazol-6-yl)oxy-
)methyl)pyridine 1-oxide; [0263]
3-(5-chloro-6-((1-methyl-1H-imidazol-2-yl)methoxy)-2-oxobenzo[d]oxazol-3(-
2H)-yl)propanoic acid; [0264]
3-(5-chloro-6-((2-methylpyridin-3-yl)methoxy)-2-oxobenzo[d]oxazol-3(2H)-y-
l)propanoic acid; [0265]
3-(5-chloro-2-oxo-6-(pyrimidin-2-ylmethoxy)benzo[d]oxazol-3(2H)-yl)propan-
oic acid; [0266]
3-(5-chloro-6-((2-methyloxazol-5-yl)methoxy)-2-oxobenzo[d]oxazol-3(2H)-yl-
)propanoic acid; [0267]
3-(5-chloro-6-((5-methyloxazol-2-yl)methoxy)-2-oxobenzo[d]oxazol-3(2H)-yl-
)propanoic acid; [0268]
3-(6-((1H-imidazol-2-yl)methoxy)-5-chloro-2-oxobenzo[d]oxazol-3(2H)-yl)pr-
opanoic acid; and [0269]
3-(5-chloro-2-oxo-6-(pyridazin-3-ylmethoxy)benzo[d]oxazol-3(2H)-yl)propan-
oic acid; [0270] and salts thereof
[0271] In a further embodiment, compounds of formula (I) or salts
thereof are selected from the group consisting of: [0272]
(R)-3-(5-chloro-2-oxo-6-(1-(pyridin-2-yl)ethoxy)benzo[d]oxazol-3(2H)-yl)p-
ropanoic acid; [0273]
3-(5-chloro-6-(1-(2-methyloxazol-5-yl)ethoxy)-2-oxobenzo[d]oxazol-3(2H)-y-
l)propanoic acid (racemic); [0274]
(R)-3-(5-chloro-6-(1-(5-fluoropyridin-2-yl)ethoxy)-2-oxobenzo[d]oxazol-3(-
2H)-yl)propanoic acid; [0275]
(R)-3-(5-chloro-6-(1-(5-methylpyridin-2-yl)ethoxy)-2-oxobenzo[d]oxazol-3(-
2H)-yl)propanoic acid; [0276]
3-(5-chloro-6-((5-fluoropyridin-2-yl)methoxy)-2-oxobenzo[d]oxazol-3(2H)-y-
l)propanoic acid; [0277]
3-(5-chloro-6-((5-chloropyridin-2-yl)methoxy)-2-oxobenzo[d]oxazol-3(2H)-y-
l)propanoic acid; [0278]
(R)-3-(5-chloro-6-(1-(5-chloropyridin-2-yl)ethoxy)-2-oxobenzo[d]oxazol-3(-
2H)-yl)propanoic acid; [0279]
3-(5-chloro-6-((5-methylpyridin-2-yl)methoxy)-2-oxobenzo[d]oxazol-3(2H)-y-
l)propanoic acid; [0280]
(R)-3-(5-chloro-6-(1-(6-methylpyridin-2-yl)ethoxy)-2-oxobenzo[d]oxazol-3(-
2H)-yl)propanoic acid; [0281]
(R)-3-(5-chloro-2-oxo-6-(1-(pyridin-2-yl)propoxy)benzo[d]oxazol-3(2H)-yl)-
propanoic acid; [0282]
(R)-3-(5-chloro-6-(1-(4-methylpyridin-2-yl)ethoxy)-2-oxobenzo[d]oxazol-3(-
2H)-yl)propanoic acid; [0283]
(R)-3-(5-chloro-2-oxo-6-(1-(pyridazin-3-yl)ethoxy)benzo[d]oxazol-3(2H)-yl-
)propanoic acid; [0284]
(S)-3-(5-chloro-2-oxo-6-(1-(pyridazin-3-yl)ethoxy)benzo[d]oxazol-3(2H)-yl-
)propanoic acid; [0285]
3-(5-chloro-6-((6-methylpyridazin-3-yl)methoxy)-2-oxobenzo[d]oxazol-3(2H)-
-yl)propanoic acid; [0286]
(R)-3-(5-chloro-6-(1-(6-methylpyridazin-3-yl)ethoxy)-2-oxobenzo[d]oxazol--
3(2H)-yl)propanoic acid; [0287]
(S)-3-(5-chloro-6-(1-(6-methylpyridazin-3-yl)ethoxy)-2-oxobenzo[d]oxazol--
3(2H)-yl)propanoic acid; [0288]
(R)-3-(5-chloro-6-(1-(5-methylpyridin-2-yl)propoxy)-2-oxobenzo[d]oxazol-3-
(2H)-yl)propanoic acid; [0289]
(R)-3-(5-chloro-6-(1-(5-methylpyridin-2-yl)ethoxy)-2-oxobenzo[d]oxazol-3(-
2H)-yl)-2-methylpropanoic acid; [0290]
(S)-3-(5-chloro-6-(1-(5-methylpyridin-2-yl)ethoxy)-2-oxobenzo[d]oxazol-3(-
2H)-yl)-2-methylpropanoic acid; [0291]
(R)-3-(5-chloro-6-(1-(5-methylpyridin-2-yl)ethoxy)-2-oxobenzo[d]oxazol-3(-
2H)-yl)-2-methylpropanoic acid; [0292]
(S)-3-(5-chloro-6-(1-(5-methylpyridin-2-yl)ethoxy)-2-oxobenzo[d]oxazol-3(-
2H)-yl)-2-methylpropanoic acid; [0293]
3-(5-chloro-2-oxo-6-(1-(pyrimidin-2-yl)ethoxy)benzo[d]oxazol-3(2H)-yl)pro-
panoic acid (single unknown enantiomer); [0294]
(R)-3-(5-chloro-6-(1-(oxazol-2-yl)ethoxy)-2-oxobenzo[d]oxazol-3(2H)-yl)pr-
opanoic acid; and [0295]
(S)-3-(5-chloro-6-(1-(oxazol-2-yl)ethoxy)-2-oxobenzo[d]oxazol-3(2H)-yl)pr-
opanoic acid; [0296] and salts thereof
[0297] The compounds of formula (I) are capable of forming base
addition salts. Such salts can be formed by reaction with the
appropriate base, optionally in a suitable solvent such as an
organic solvent, to give the salt which can be isolated by
crystallisation and filtration.
[0298] Because of their potential use in medicine, it will be
appreciated that for use in medicine the salts of the compounds of
formula (I) should be pharmaceutically acceptable. Suitable
pharmaceutically acceptable salts will be apparent to those skilled
in the art and include those described in Berge, J. Pharm. Sci.,
1977, 66, 1-19. Pharmaceutically acceptable base salts include, but
are not limited to, ammonium salts, alkali metal salts such as
those of sodium and potassium, alkaline earth metal salts such as
those of calcium and magnesium and salts with organic bases,
including salts of primary, secondary and tertiary amines, such as
t-butylamine, cyclohexylamine, dimethylamine, trimethylamine,
diethyltriamine, 2-amino-2-(hydroxymethyl)-1,3-propanediol (TRIS)
and N-methyl-D-glucamine.
[0299] In a further embodiment, compounds of formula (I) or salts
thereof are selected from the group consisting of: [0300]
3-(5-chloro-6-methyl-2-oxobenzo[d]oxazol-3(2H)-yl)propanoic acid;
[0301] 2-amino-2-(hydroxymethyl)propane-1,3-diol
3-(5-chloro-6-methyl-2-oxobenzo[d]oxazol-3(2H)-yl)propanoate;
[0302]
3-(5-chloro-2-oxo-6-(1-(pyridin-2-yl)ethoxy)benzo[d]oxazol-3(2H)-yl)propa-
noic acid; [0303]
(S)-3-(5-chloro-2-oxo-6-(1-(pyridin-2-yl)ethoxy)benzo[d]oxazol-3(2H)-yl)p-
ropanoic acid; [0304] 2-amino-2-(hydroxymethyl)propane-1,3-diol
(S)-3-(5-chloro-2-oxo-6-(1-(pyridin-2-yl)ethoxy)benzo[d]oxazol-3(2H)-yl)p-
ropanoate; [0305]
(R)-3-(5-chloro-2-oxo-6-(1-(pyridin-2-yl)ethoxy)benzo[d]oxazol-3(2H)-yl)p-
ropanoic acid; [0306] 2-amino-2-(hydroxymethyl)propane-1,3-diol
(R)-3-(5-chloro-2-oxo-6-(1-(pyridin-2-yl)ethoxy)benzo[d]oxazol-3(2H)-yl)p-
ropanoate; [0307]
3-(5,6-dichloro-2-oxobenzo[d]oxazol-3(2H)-yl)propanoic acid; [0308]
3-(5-chloro-6-fluoro-2-oxobenzo[d]oxazol-3(2H)-yl)propanoic acid;
[0309] 3-(5-chloro-6-methoxy-2-oxobenzo[d]oxazol-3(2H)-yl)propanoic
acid; [0310]
3-(5-chloro-6-cyano-2-oxobenzo[d]oxazol-3(2H)-yl)propanoic acid;
[0311] 2-amino-2-(hydroxymethyl)propane-1,3-diol
3-(6-bromo-5-chloro-2-oxobenzo[d]oxazol-3(2H)-yl)propanoate; [0312]
2-amino-2-(hydroxymethyl)propane-1,3-diol
3-(5-chloro-6-ethoxy-2-oxobenzo[d]oxazol-3(2H)-yl)propanoate;
[0313] 2-amino-2-(hydroxymethyl)propane-1,3-diol
3-(5-chloro-6-ethyl-2-oxobenzo[d]oxazol-3(2H)-yl)propanoate; [0314]
2-amino-2-(hydroxymethyl)propane-1,3-diol
3-(5-chloro-6-isobutoxy-2-oxobenzo[d]oxazol-3(2H)-yl)propanoate;
[0315] 2-amino-2-(hydroxymethyl)propane-1,3-diol
3-(5-chloro-6-(cyclopropylmethoxy)-2-oxobenzo[d]oxazol-3(2H)-yl)propanoat-
e; [0316] 2-amino-2-(hydroxymethyl)propane-1,3-diol
3-(5-chloro-6-(cyclobutylmethoxy)-2-oxobenzo[d]oxazol-3(2H)-yl)propanoate-
; [0317] 2-amino-2-(hydroxymethyl)propane-1,3-diol
3-(6-(benzyloxy)-5-chloro-2-oxobenzo[d]oxazol-3(2H)-yl)propanoate;
[0318] 2-amino-2-(hydroxymethyl)propane-1,3-diol
3-(5-chloro-6-(2-methoxyethoxy)-2-oxobenzo[d]oxazol-3(2H)-yl)propanoate;
[0319] 2-amino-2-(hydroxymethyl)propane-1,3-diol
3-(5-chloro-2-oxo-6-propoxybenzo[d]oxazol-3(2H)-yl)propanoate;
[0320] 2-amino-2-(hydroxymethyl)propane-1,3-diol
3-(5-chloro-2-oxo-6-(3,3,3-trifluoropropoxy)benzo[d]oxazol-3(2H)-yl)propa-
noate; [0321] 2-amino-2-(hydroxymethyl)propane-1,3-diol
3-(5-chloro-2-oxo-6-(pyridin-2-ylmethoxy)benzo[d]oxazol-3(2H)-yl)propanoa-
te; [0322] 2-amino-2-(hydroxymethyl)propane-1,3-diol
3-(5-chloro-6-isopropoxy-2-oxobenzo[d]oxazol-3(2H)-yl)propanoate;
[0323] 2-amino-2-(hydroxymethyl)propane-1,3-diol
3-(5-chloro-6-cyclobutoxy-2-oxobenzo[d]oxazol-3(2H)-yl)propanoate;
[0324] 2-amino-2-(hydroxymethyl)propane-1,3-diol
3-(5-chloro-6-cyclopropoxy-2-oxobenzo[d]oxazol-3(2H)-yl)propanoate;
[0325] 2-amino-2-(hydroxymethyl)propane-1,3-diol
3-(5-chloro-2-oxo-6-(2,2,2-trifluoroethoxy)benzo[d]oxazol-3(2H)-yl)propan-
oate; [0326] 2-amino-2-(hydroxymethyl)propane-1,3-diol
3-(5-chloro-6-(oxetan-3-yloxy)-2-oxobenzo[d]oxazol-3(2H)-yl)propanoate;
[0327] 2-amino-2-(hydroxymethyl)propane-1,3-diol
3-(5-chloro-6-isobutyl-2-oxobenzo[d]oxazol-3(2H)-yl)propanoate;
[0328] 2-amino-2-(hydroxymethyl)propane-1,3-diol
3-(5-chloro-6-(oxetan-3-ylmethoxy)-2-oxobenzo[d]oxazol-3(2H)-yl)propanoat-
e; [0329] 2-amino-2-(hydroxymethyl)propane-1,3-diol
3-(5-chloro-6-((1-methoxypropan-2-yl)oxy)-2-oxobenzo[d]oxazol-3(2H)-yl)pr-
opanoate; [0330] 2-amino-2-(hydroxymethyl)propane-1,3-diol
3-(5-chloro-6-((6-methylpyridin-3-yl)methoxy)-2-oxobenzo[d]oxazol-3(2H)-y-
l)propanoate; [0331] 2-amino-2-(hydroxymethyl)propane-1,3-diol
3-(5-chloro-6-((2-methylpyridin-4-yl)methoxy)-2-oxobenzo[d]oxazol-3(2H)-y-
l)propanoate; [0332]
2-(((3-(2-carboxyethyl)-5-chloro-2-oxo-2,3-dihydrobenzo[d]oxazol-6-yl)oxy-
)methyl)pyridine 1-oxide; [0333]
2-amino-2-(hydroxymethyl)propane-1,3-diol
3-(5-chloro-6-((1-methyl-1H-imidazol-2-yl)methoxy)-2-oxobenzo[d]oxazol-3(-
2H)-yl)propanoate; [0334] 2-amino-2-(hydroxymethyl)propane-1,3-diol
3-(5-chloro-6-((2-methylpyridin-3-yl)methoxy)-2-oxobenzo[d]oxazol-3(2H)-y-
l)propanoate; [0335] 2-amino-2-(hydroxymethyl)propane-1,3-diol
3-(5-chloro-2-oxo-6-(pyrimidin-2-ylmethoxy)benzo[d]oxazol-3(2H)-yl)propan-
oate; [0336] 2-amino-2-(hydroxymethyl)propane-1,3-diol
3-(5-chloro-6-((2-methyloxazol-5-yl)methoxy)-2-oxobenzo[d]oxazol-3(2H)-yl-
)propanoate; [0337] 2-amino-2-(hydroxymethyl)propane-1,3-diol
3-(5-chloro-6-((5-methyloxazol-2-yl)methoxy)-2-oxobenzo[d]oxazol-3(2H)-yl-
)propanoate; [0338]
3-(6-((1H-imidazol-2-yl)methoxy)-5-chloro-2-oxobenzo[d]oxazol-3(2H)-yl)pr-
opanoic acid; and [0339] 2-amino-2-(hydroxymethyl)propane-1,3-diol
3-(5-chloro-2-oxo-6-(pyridazin-3-ylmethoxy)benzo[d]oxazol-3(2H)-yl)propan-
oate.
[0340] In a further embodiment, compounds of formula (I) or salts
thereof are selected from the group consisting of: [0341]
(R)-3-(5-chloro-2-oxo-6-(1-(pyridin-2-yl)ethoxy)benzo[d]oxazol-3(2H)-yl)p-
ropanoic acid hydrochloride; [0342]
(R)-3-(5-chloro-2-oxo-6-(1-(pyridin-2-yl)ethoxy)benzo[d]oxazol-3(2H)-yl)p-
ropanoic acid compound with sulfuric acid (1:1); [0343]
(R)-3-(5-chloro-2-oxo-6-(1-(pyridin-2-yl)ethoxy)benzo[d]oxazol-3(2H)-yl)p-
ropanoic acid compound with methanesulfonic acid (1:1); [0344]
N-benzyl-2-phenylethanamine
(R)-3-(5-chloro-2-oxo-6-(1-(pyridin-2-yl)ethoxy)benzo[d]oxazol-3(2H)-yl)p-
ropanoate; [0345] N1,N2-dibenzylethane-1,2-diamine
(R)-3-(5-chloro-2-oxo-6-(1-(pyridin-2-yl)ethoxy)benzo[d]oxazol-3(2H)-yl)p-
ropanoate; [0346]
(2R,3R,4S,5R)-3,4,5,6-tetrahydroxy-2-(methylamino)hexanal
(R)-3-(5-chloro-2-oxo-6-(1-(pyridin-2-yl)ethoxy)benzo[d]oxazol-3(2H)-yl)p-
ropanoate; [0347]
(R)-3-(5-chloro-2-oxo-6-(1-(pyridin-2-yl)ethoxy)benzo[d]oxazol-3(2H)-yl)p-
ropanoic acid compound with (S)-2-amino-5-guanidinopentanoic acid
(1:1); [0348]
(R)-3-(5-chloro-2-oxo-6-(1-(pyridin-2-yl)ethoxy)benzo[d]oxazol-3(2-
H)-yl)propanoic acid compound with (S)-2,6-diaminohexanoic acid
(1:1); [0349] sodium
(R)-3-(5-chloro-2-oxo-6-(1-(pyridin-2-yl)ethoxy)benzo[d]oxazol-3(2H)-yl)p-
ropanoate; [0350]
(R)-3-(5-chloro-2-oxo-6-(1-(pyridin-2-yl)ethoxy)benzo[d]oxazol-3(2H)-yl)p-
ropanoic acid compound with 4-methylbenzenesulfonic acid (1:1);
[0351] N1-(2-aminoethyl)ethane-1,2-diamine
(R)-3-(5-chloro-2-oxo-6-(1-(pyridin-2-yl)ethoxy)benzo[d]oxazol-3(2H)-yl)p-
ropanoate; [0352] 2-amino-2-(hydroxymethyl)propane-1,3-diol
3-(5-chloro-6-(1-(2-methyloxazol-5-yl)ethoxy)-2-oxobenzo[d]oxazol-3(2H)-y-
l)propanoate (racemic); [0353]
2-amino-2-(hydroxymethyl)propane-1,3-diol
(R)-3-(5-chloro-6-(1-(5-fluoropyridin-2-yl)ethoxy)-2-oxobenzo[d]oxazol-3(-
2H)-yl)propanoate; [0354] 2-amino-2-(hydroxymethyl)propane-1,3-diol
(R)-3-(5-chloro-6-(1-(5-methylpyridin-2-yl)ethoxy)-2-oxobenzo[d]oxazol-3(-
2H)-yl)propanoate; [0355]
(R)-3-(5-chloro-6-(1-(5-methylpyridin-2-yl)ethoxy)-2-oxobenzo[d]oxazol-3(-
2H)-yl)propanoic acid; [0356]
2-amino-2-(hydroxymethyl)propane-1,3-diol
3-(5-chloro-6-((5-fluoropyridin-2-yl)methoxy)-2-oxobenzo[d]oxazol-3(2H)-y-
l)propanoate; [0357] 2-amino-2-(hydroxymethyl)propane-1,3-diol
3-(5-chloro-6-((5-chloropyridin-2-yl)methoxy)-2-oxobenzo[d]oxazol-3(2H)-y-
l)propanoate; [0358] 2-amino-2-(hydroxymethyl)propane-1,3-diol
(R)-3-(5-chloro-6-(1-(5-chloropyridin-2-yl)ethoxy)-2-oxobenzo[d]oxazol-3(-
2H)-yl)propanoate; [0359]
(R)-3-(5-chloro-6-(1-(5-chloropyridin-2-yl)ethoxy)-2-oxobenzo[d]oxazol-3(-
2H)-yl)propanoic acid; [0360]
2-amino-2-(hydroxymethyl)propane-1,3-diol
3-(5-chloro-6-((5-methylpyridin-2-yl)methoxy)-2-oxobenzo[d]oxazol-3(2H)-y-
l)propanoate; [0361] 2-amino-2-(hydroxymethyl)propane-1,3-diol
(R)-3-(5-chloro-6-(1-(6-methylpyridin-2-yl)ethoxy)-2-oxobenzo[d]oxazol-3(-
2H)-yl)propanoate; [0362] 2-amino-2-(hydroxymethyl)propane-1,3-diol
(R)-3-(5-chloro-2-oxo-6-(1-(pyridin-2-yl)propoxy)benzo[d]oxazol-3(2H)-yl)-
propanoate; [0363]
(R)-3-(5-chloro-2-oxo-6-(1-(pyridin-2-yl)propoxy)benzo[d]oxazol-3(2H)-yl)-
propanoic acid; [0364] 2-amino-2-(hydroxymethyl)propane-1,3-diol
(R)-3-(5-chloro-6-(1-(4-methylpyridin-2-yl)ethoxy)-2-oxobenzo[d]oxazol-3(-
2H)-yl)propanoate; [0365]
(R)-3-(5-chloro-2-oxo-6-(1-(pyridazin-3-yl)ethoxy)benzo[d]oxazol-3(2H)-yl-
)propanoic acid; [0366]
(S)-3-(5-chloro-2-oxo-6-(1-(pyridazin-3-yl)ethoxy)benzo[d]oxazol-3(2H)-yl-
)propanoic acid; [0367]
3-(5-chloro-6-((6-methylpyridazin-3-yl)methoxy)-2-oxobenzo[d]oxazol-3(2H)-
-yl)propanoic acid; [0368]
(R)-3-(5-chloro-6-(1-(6-methylpyridazin-3-yl)ethoxy)-2-oxobenzo[d]oxazol--
3(2H)-yl)propanoic acid; [0369]
(S)-3-(5-chloro-6-(1-(6-methylpyridazin-3-yl)ethoxy)-2-oxobenzo[d]oxazol--
3(2H)-yl)propanoic acid; [0370]
(R)-3-(5-chloro-6-(1-(5-methylpyridin-2-yl)propoxy)-2-oxobenzo[d]oxazol-3-
(2H)-yl)propanoic acid; [0371]
(R)-3-(5-chloro-6-(1-(5-methylpyridin-2-yl)ethoxy)-2-oxobenzo[d]oxazol-3(-
2H)-yl)-2-methylpropanoic acid; [0372]
(S)-3-(5-chloro-6-(1-(5-methylpyridin-2-yl)ethoxy)-2-oxobenzo[d]oxazol-3(-
2H)-yl)-2-methylpropanoic acid; [0373]
(R)-3-(5-chloro-6-(1-(5-methylpyridin-2-yl)ethoxy)-2-oxobenzo[d]oxazol-3(-
2H)-yl)-2-methylpropanoic acid; [0374]
(S)-3-(5-chloro-6-(1-(5-methylpyridin-2-yl)ethoxy)-2-oxobenzo[d]oxazol-3(-
2H)-yl)-2-methylpropanoic acid; [0375]
2-amino-2-(hydroxymethyl)propane-1,3-diol
3-(5-chloro-2-oxo-6-(1-(pyrimidin-2-yl)ethoxy)benzo[d]oxazol-3(2H)-yl)pro-
panoate (single unknown enantiomer); [0376]
2-amino-2-(hydroxymethyl)propane-1,3-diol
(S)-3-(5-chloro-6-(1-(oxazol-2-yl)ethoxy)-2-oxobenzo[d]oxazol-3(2H)-yl)pr-
opanoate; and [0377] 2-amino-2-(hydroxymethyl)propane-1,3-diol
(R)-3-(5-chloro-6-(1-(oxazol-2-yl)ethoxy)-2-oxobenzo[d]oxazol-3(2H)-yl)pr-
opanoate.
[0378] In one embodiment, the compound of formula (I) or a salt
thereof is: [0379]
3-(5-chloro-6-methyl-2-oxobenzo[d]oxazol-3(2H)-yl)propanoic acid or
a salt thereof.
[0380] In a particular embodiment, the compound of formula (I) or a
salt thereof is: [0381]
3-(5-chloro-6-methyl-2-oxobenzo[d]oxazol-3(2H)-yl)propanoic acid or
a pharmaceutically acceptable salt thereof.
[0382] In a more particular embodiment, the compound of formula (I)
or a salt thereof is: [0383]
3-(5-chloro-6-methyl-2-oxobenzo[d]oxazol-3(2H)-yl)propanoic acid
2-amino-2-(hydroxymethyl)-1,3-propanediol salt.
[0384] In an alternative embodiment, the compound of formula (I) or
a salt thereof is:
3-(5-chloro-6-methyl-2-oxobenzo[d]oxazol-3(2H)-yl)propanoic
acid.
[0385] In an alternative embodiment, the compound of formula (I) or
a salt thereof is
3-(5-chloro-2-oxo-6-(1-(pyridin-2-yl)ethoxy)benzo[d]oxazol-3(2H)-yl)propa-
noic acid or a salt thereof.
[0386] In one embodiment, the compound of formula (I) or a salt
thereof is
(S)-3-(5-chloro-2-oxo-6-(1-(pyridin-2-yl)ethoxy)benzo[d]oxazol-3(2H)-yl)p-
ropanoic acid or a salt thereof.
[0387] In one embodiment, the compound of formula (I) or a salt
thereof is
(R)-3-(5-chloro-2-oxo-6-(1-(pyridin-2-yl)ethoxy)benzo[d]oxazol-3(2H)-yl)p-
ropanoic acid or a salt thereof.
[0388] In a particular embodiment, the compound of formula (I) or a
salt thereof is
3-(5-chloro-2-oxo-6-(1-(pyridin-2-yl)ethoxy)benzo[d]oxazol-3(2H)-yl)propa-
noic acid or a pharmaceutically acceptable salt thereof.
[0389] In a particular embodiment, the compound of formula (I) or a
salt thereof is
(S)-3-(5-chloro-2-oxo-6-(1-(pyridin-2-yl)ethoxy)benzo[d]oxazol-3(2H)-yl)p-
ropanoic acid or a pharmaceutically acceptable salt thereof.
[0390] In a particular embodiment, the compound of formula (I) or a
salt thereof is
(R)-3-(5-chloro-2-oxo-6-(1-(pyridin-2-yl)ethoxy)benzo[d]oxazol-3(2H)-yl)p-
ropanoic acid or a pharmaceutically acceptable salt thereof.
[0391] In a more particular embodiment, the compound of formula (I)
or a salt thereof is
3-(5-chloro-2-oxo-6-(1-(pyridin-2-yl)ethoxy)benzo[d]oxazol-3(2H)-yl)propa-
noic acid 2-amino-2-(hydroxymethyl)-1,3-propanediol salt.
[0392] In a more particular embodiment the compound of formula (I)
or a salt thereof is
(R)-3-(5-chloro-2-oxo-6-(1-(pyridin-2-yl)ethoxy)benzo[d]oxazol-3(2H)-yl)p-
ropanoic acid 2-amino-2-(hydroxymethyl)-1,3-propanediol salt.
[0393] In an alternative embodiment the compound of formula (I) or
a salt thereof is
(S)-3-(5-chloro-2-oxo-6-(1-(pyridin-2-yl)ethoxy)benzo[d]oxazol-3(2H)-yl)p-
ropanoic acid 2-amino-2-(hydroxymethyl)-1,3-propanediol salt.
[0394] The invention includes within its scope all possible
stoichiometric and non-stoichiometric forms of the salts for the
compounds of formula (I).
[0395] Certain compounds of formula (I) or salts thereof may exist
in the form of solvates. As used herein, the term "solvate" refers
to a complex of variable stoichiometry formed by a solute (in this
invention, a compound of formula (I) or a salt thereof) and a
solvent. Such solvents for the purpose of the invention may not
interfere with the biological activity of the solute. Examples of
suitable solvents include water, methanol, ethanol and acetic acid.
If the solvent used is water, the solvate may be referred to as a
hydrate.
[0396] Certain compounds of formula (I) are capable of existing in
sterioisomeric forms. It will be understood that the invention
encompasses all geometric and optical isomers of these compounds
and the mixtures thereof including racemates. The different
stereoisomeric forms may be separated one from the other by methods
known in the art (i.e. separation by chiral HPLC) or any given
isomer may be obtained by stereospecific or asymmetric synthesis.
The invention also extends to any tautomeric forms and mixtures
thereof.
[0397] The invention also includes isotopically-labelled compounds
and salts, which are identical to compounds of formula (I) or salts
thereof, but for the fact that one or more atoms are replaced by an
atom having an atomic mass or mass number different from the atomic
mass or mass number most commonly found in nature. Examples of
isotopes that can be incorporated into compounds of formula (I) or
salts thereof isotopes of hydrogen, carbon, nitrogen, fluorine,
such as .sup.3H, .sup.11C, .sup.14C and .sup.18F. Such
isotopically-labelled compound of formula (I) or salts thereof are
useful in drug and/or substrate tissue distribution assays. For
example, .sup.11C and .sup.18F isotopes are particularly useful in
PET (positron emission tomography). PET is useful in brain imaging.
Isotopically labelled compounds of formula (I) and salts thereof
can generally be prepared by carrying out the procedures disclosed
below, by substituting a readily available isotopically labelled
reagent for a non-isotopically labelled reagent. In one embodiment,
compounds of formula (I) or salts thereof are not isotopically
labelled.
[0398] In order to use the compounds of formula (I) in therapy,
they will normally be formulated into a pharmaceutical composition
in accordance with standard pharmaceutical practice.
[0399] In a further aspect, the invention provides a pharmaceutical
composition, which comprises a) a compound for formula (I) or a
pharmaceutically acceptable salt thereof; and b) one or more
pharmaceutically acceptable excipients.
[0400] In one embodiment, the compound of formula (I) or a
pharmaceutically acceptable salt thereof is
3-(5-chloro-6-methyl-2-oxobenzo[d]oxazol-3(2H)-yl)propanoic acid or
a pharmaceutically acceptable salts thereof.
[0401] In a further aspect the invention provides a pharmaceutical
composition, which comprises a compound of formula (I) or a
pharmaceutically acceptable salt thereof, and a pharmaceutically
acceptable carrier or excipient.
[0402] In a further aspect, the present invention provides a
process for preparing a pharmaceutical composition, the process
comprising mixing a compound of formula (I) or a pharmaceutically
acceptable salt thereof and a pharmaceutically acceptable carrier
or excipient.
[0403] A pharmaceutical composition of the invention, which may be
prepared by admixture, suitably at ambient temperature and
atmospheric pressure, is usually adapted for oral, parenteral or
rectal administration and, as such, may be in the form of tablets,
capsules, oral liquid preparations, powders, granules, lozenges,
reconstitutable powders, injectable or infusible solutions or
suspensions or suppositories.
[0404] In one embodiment injectable or infusible solutions, or
reconstitutable powders, are preferred.
[0405] In an alternative embodiment, a composition adapted for oral
formulation is preferred.
[0406] Tablets and capsules for oral administration may be in unit
dose form, and may contain conventional excipients, such as binding
agents (e.g. pregelatinised maize starch, polyvinylpyrrolidone or
hydroxypropyl methylcellulose); fillers (e.g. lactose,
microcrystalline cellulose or calcium hydrogen phosphate);
tabletting lubricants (e.g. magnesium stearate, talc or silica);
disintegrants (e.g. potato starch or sodium starch glycollate); and
acceptable wetting agents (e.g. sodium lauryl sulphate). The
tablets may be coated according to methods well known in normal
pharmaceutical practice.
[0407] Oral liquid preparations may be in the form of, for example,
aqueous or oily suspension, solutions, emulsions, syrups or
elixirs, or may be in the form of a dry product for reconstitution
with water or other suitable vehicle before use. Such liquid
preparations may contain conventional additives such as suspending
agents (e.g. sorbitol syrup, cellulose derivatives or hydrogenated
edible fats), emulsifying agents (e.g. lecithin or acacia),
non-aqueous vehicles (which may include edible oils e.g. almond
oil, oily esters, ethyl alcohol or fractionated vegetable oils),
preservatives (e.g. methyl or propyl-p-hydroxybenzoates or sorbic
acid), and, if desired, conventional flavourings or colorants,
buffer salts and sweetening agents as appropriate. Preparations for
oral administration may be suitably formulated to give controlled
release of the active compound.
[0408] For parenteral administration, fluid unit dosage forms are
prepared utilising a compound of the invention or pharmaceutically
acceptable salt thereof and a sterile vehicle. Formulations for
injection may be presented in unit dosage form e.g. in ampoules or
in multi-dose, utilising a compound of the invention or
pharmaceutically acceptable salt thereof and a sterile vehicle,
optionally with an added preservative. The compositions may take
such forms as suspensions, solutions or emulsions in oily or
aqueous vehicles, and may contain formulatory agents such as
suspending, stabilising and/or dispersing agents. Alternatively,
the active ingredient may be in powder form for constitution with a
suitable vehicle, e.g. sterile pyrogen-free water, before use. The
compound, depending on the vehicle and concentration used, can be
either suspended or dissolved in the vehicle. In preparing
solutions, the compound can be dissolved for injection and filter
sterilised before filling into a suitable vial or ampoule and
sealing. Advantageously, adjuvants such as a local anaesthetic,
preservatives and buffering agents are dissolved in the vehicle. To
enhance the stability, the composition can be frozen after filling
into the vial and the water removed under vacuum. Parenteral
suspensions are prepared in substantially the same manner, except
that the compound is suspended in the vehicle instead of being
dissolved, and sterilisation cannot be accomplished by filtration.
The compound can be sterilised by exposure to ethylene oxide before
suspension in a sterile vehicle. Advantageously, a surfactant or
wetting agent is included in the composition to facilitate uniform
distribution of the compound.
[0409] Lotions may be formulated with an aqueous or oily base and
will in general also contain one or more emulsifying agents,
stabilising agents, dispersing agents, suspending agents,
thickening agents, or colouring agents. Drops may be formulated
with an aqueous or non-aqueous base also comprising one or more
dispersing agents, stabilising agents, solubilising agents or
suspending agents. They may also contain a preservative.
[0410] The compounds of the invention may also be formulated in
rectal compositions such as suppositories or retention enemas, e.g.
containing conventional suppository bases such as cocoa butter or
other glycerides.
[0411] The compounds of the invention may also be formulated as
depot preparations. Such long acting formulations may be
administered by implantation (for example subcutaneously or
intramuscularly) or by intramuscular injection. Thus, for example,
the compounds of the invention may be formulated with suitable
polymeric or hydrophobic materials (for example as an emulsion in
an acceptable oil) or ion exchange resins, or as sparingly soluble
derivatives, for example, as a sparingly soluble salt.
[0412] For intranasal administration, the compounds of the
invention may be formulated as solutions for administration via a
suitable metered or unitary dose device or alternatively as a
powder mix with a suitable carrier for administration using a
suitable delivery device. Thus compounds of formula (I) may be
formulated for oral, buccal, parenteral, topical (including
ophthalmic and nasal), depot or rectal administration or in a form
suitable for administration by inhalation or insufflation (either
through the mouth or nose).
[0413] The compounds of the invention may be formulated for topical
administration in the form of ointments, creams, gels, lotions,
pessaries, aerosols or drops (e.g. eye, ear or nose drops).
Ointments and creams may, for example, be formulated with an
aqueous or oily base with the addition of suitable thickening
and/or gelling agents. Ointments for administration to the eye may
be manufactured in a sterile manner using sterilised
components.
[0414] The composition may contain from 0.1% to 99% by weight,
preferably from 10 to 60% by weight, of the active material,
depending on the method of administration. The dose of the compound
used in the treatment of the aforementioned disorders will vary in
the usual way with the seriousness of the disorders, the weight of
the sufferer, and other similar factors. However, as a general
guide suitable unit doses may be 0.05 to 5000 mg, 1.0 to 500 mg or
1.0 to 200 mg and such unit doses may be administered more than
once a day, for example two or three times a day. Such therapy may
extend for a number of weeks, months or years.
[0415] Compounds of formula (I) and their pharmaceutically
acceptable salts are therefore of use in the treatment of
conditions or disorders which are mediated via KMO. In particular
the compounds of formula (I) and their pharmaceutically acceptable
salts are of use in the treatment of acute pancreatitis, chronic
kidney disease, other conditions associated with systemic
inflammatory response syndrome (SIRS), Huntington's disease,
Alzheimer's disease, spinocerebellar ataxias, Parkinson's disease,
AIDS-dementia complex, amylotrophic lateral sclerosis (ALS),
depression, schizophrenia, sepsis, cardiovascular shock, severe
trauma, acute lung injury, acute respiratory distress syndrome,
acute cholecystitis, severe burns, pneumonia, extensive surgical
procedures, ischemic bowel, severe acute hepatic disease, severe
acute hepatic encephalopathy or acute renal failure.
[0416] In one embodiment, the compounds of formula (I) and their
pharmaceutically acceptable salts are of use in the treatment of
acute pancreatitis, other conditions associated with systemic
inflammatory response syndrome (SIRS), Huntington's disease,
Alzheimer's disease, spinocerebellar ataxias, Parkinson's disease,
AIDS-dementia complex, amylotrophic lateral sclerosis (ALS),
depression, schizophrenia, sepsis, cardiovascular shock, severe
trauma, acute lung injury, acute respiratory distress syndrome,
acute cholecystitis, severe burns, pneumonia, extensive surgical
procedures, ischemic bowel, severe acute hepatic disease, severe
acute hepatic encephalopathy or acute renal failure.
[0417] It is to be understood that "treatment" as used herein
includes prophylaxis as well as alleviation of established
symptoms, signs and features of disease.
[0418] In a further aspect, the invention provides a compound of
formula (I) or a pharmaceutically acceptable salt thereof for use
in therapy.
[0419] The invention further provides a method of treatment of
conditions or disorders in mammals including humans which can be
mediated via KMO, which method comprises administering to the
sufferer a therapeutically safe and effective amount of a compound
of formula (I) or a pharmaceutically acceptable salt thereof.
[0420] The invention further provides a method of treatment of
acute pancreatitis, other conditions associated with systemic
inflammatory response syndrome (SIRS), Huntington's disease,
Alzheimer's disease, spinocerebellar ataxias, Parkinson's disease,
AIDS-dementia complex, amylotrophic lateral sclerosis (ALS),
depression, schizophrenia, sepsis, cardiovascular shock, severe
trauma, acute lung injury, acute respiratory distress syndrome,
acute cholecystitis, severe burns, pneumonia, extensive surgical
procedures, ischemic bowel, severe acute hepatic disease, severe
acute hepatic encephalopathy or acute renal failure which method
comprises administering to the sufferer a therapeutically safe and
effective amount of a compound of formula (I) or a pharmaceutically
acceptable salt thereof.
[0421] The invention further provides a method of treatment of
acute pancreatitis, which method comprises administering to the
sufferer a therapeutically safe and effective amount of a compound
of formula (I) or a pharmaceutically acceptable salt thereof.
[0422] The invention further provides a method of treatment of
chronic kidney disease, which method comprises administering to the
sufferer a therapeutically safe and effective amount of a compound
of formula (I) or a pharmaceutically acceptable salt thereof.
[0423] In another aspect, the invention provides for the use of a
compound of formula (I) or a pharmaceutically acceptable salt
thereof in the manufacture of a medicament for use in the treatment
of conditions or disorders mediated via KMO.
[0424] In another aspect, the invention provides for the use of a
compound of formula (I) or a pharmaceutically acceptable salt
thereof in the manufacture of a medicament for use in the treatment
of acute pancreatitis, other conditions associated with systemic
inflammatory response syndrome (SIRS), Huntington's disease,
Alzheimer's disease, spinocerebellar ataxias, Parkinson's disease,
AIDS-dementia complex, amylotrophic lateral sclerosis (ALS),
depression, schizophrenia, sepsis, cardiovascular shock, severe
trauma, acute lung injury, acute respiratory distress syndrome,
acute cholecystitis, severe burns, pneumonia, extensive surgical
procedures, ischemic bowel, severe acute hepatic disease, severe
acute hepatic encephalopathy or acute renal failure.
[0425] In another aspect, the invention provides for the use of a
compound of formula (I) or a pharmaceutically acceptable salt
thereof in the manufacture of a medicament for use in the treatment
of acute pancreatitis.
[0426] In another aspect, the invention provides for the use of a
compound of formula (I) or a pharmaceutically acceptable salt
thereof in the manufacture of a medicament for use in the treatment
of chronic kidney disease.
[0427] In another aspect, the invention provides for the compound
of formula (I) or a pharmaceutically acceptable salt thereof for
use in the treatment of conditions or disorders mediated via
KMO.
[0428] In another aspect, the invention provides for the compound
of formula (I) or a pharmaceutically acceptable salt thereof for
use in the treatment of acute pancreatitis, other conditions
associated with systemic inflammatory response syndrome (SIRS),
Huntington's disease, Alzheimer's disease, spinocerebellar ataxias,
Parkinson's disease, AIDS-dementia complex, amylotrophic lateral
sclerosis (ALS), depression, schizophrenia, sepsis, cardiovascular
shock, severe trauma, acute lung injury, acute respiratory distress
syndrome, acute cholecystitis, severe burns, pneumonia, extensive
surgical procedures, ischemic bowel, severe acute hepatic disease,
severe acute hepatic encephalopathy or acute renal failure.
[0429] In another aspect, the invention provides for the compound
of formula (I) or a pharmaceutically acceptable salt thereof for
use in the treatment of acute pancreatitis.
[0430] In another aspect, the invention provides for the compound
of formula (I) or a pharmaceutically acceptable salt thereof for
use in the treatment of chronic kidney disease.
[0431] In another aspect, the invention provides for a
pharmaceutical composition for use in the treatment of acute
pancreatitis, chronic kidney disease, other conditions associated
with systemic inflammatory response syndrome (SIRS), Huntington's
disease, Alzheimer's disease, spinocerebellar ataxias, Parkinson's
disease, AIDS-dementia complex, amylotrophic lateral sclerosis
(ALS), depression, schizophrenia, sepsis, cardiovascular shock,
severe trauma, acute lung injury, acute respiratory distress
syndrome, acute cholecystitis, severe burns, pneumonia, extensive
surgical procedures, ischemic bowel, severe acute hepatic disease,
severe acute hepatic encephalopathy or acute renal failure which
comprises a compound of formula (I) or a pharmaceutically
acceptable salt thereof and one or more pharmaceutically acceptable
excipients.
[0432] Compounds of formula (I) wherein X is a bond or --CH.sub.2--
may be synthesised substantially according to Reaction Scheme
1.
##STR00008##
[0433] Compounds of formula (I) wherein X is --O-- may be
synthesised substantially according to Reaction Scheme 2.
##STR00009## ##STR00010##
[0434] Methyl
3-(6-bromo-5-chloro-2-oxobenzo[d]oxazol-3(2H)-yl)propanoate (II)
can be used to synthesise compounds of formula (I) wherein X is
bond or --CH.sub.2--.
[0435] It will be appreciated by those skilled in the art that it
may be necessary to protect certain reactive substituents during
some of the above procedures. Standard protection and deprotection
techniques, such as those described in Greene T.W. Protective
groups in organic synthesis, New York, Wiley (1981), can be used.
For example, primary amines can be protected as phthalimide,
trifluoroacetyl, benzyl, tert-butyloxycarbonyl, benzyloxycarbonyl
or trityl derivatives. Carboxylic acid groups can be protected as
esters. Aldehyde or ketone groups can be protected as acetals,
ketals, thioacetals or thioketals. Deprotection of such groups is
achieved using conventional procedures well known in the art. For
example, protecting groups such as tert-butyloxycarbonyl may be
removed using an acid such as hydrochloric or trifluroroacetic acid
in a suitable solvent such as dichloromethane, diethylether,
1,4-dioxane, isopropanol or mixtures thereof.
[0436] Certain compounds of formula (I) are also commercially
available, including
3-(5-chloro-2-oxobenzo[d]oxazol-3(2H)-yl)propanoic acid.
EXAMPLES
[0437] The following Examples illustrate the invention. These
Examples are not intended to limit the scope of the present
invention, but rather to provide guidance to the skilled artisan to
prepare and use the compounds, compositions, and methods of the
present invention. While particular embodiments of the present
invention are described, the skilled artisan will appreciate that
various changes and modifications can be made without departing
from the spirit and scope of the invention.
[0438] All publications, including but not limited to patents and
patent applications, cited in this specification are herein
incorporated by reference as if each individual publication were
specifically and individually indicated to be incorporated by
reference herein as though fully set forth.
[0439] The following Intermediates and Examples illustrate the
preparation of compounds of the invention.
ABBREVIATIONS
[0440] DCM dichloromethane
[0441] DEAD diethyl azodicarboxylate
[0442] DMSO dimethylsulphoxide
[0443] h hour(s)
[0444] LCMS liquid chromatography-mass spectrometry
[0445] MeCN acetonitrile
[0446] min minutes
[0447] NMR nuclear magnetic resonance
[0448] RT/Rt: retention time
[0449] THF tetrahydrofuran
[0450] TFA trifluoroacetic acid
[0451] TRIS 2-amino-2-(hydroxymethyl)-1,3-propanediol
[0452] LCMS Conditions
[0453] LCMS Method A/B/C
[0454] Agilent 1200-6110,
[0455] Signal table: Signal A: 214 nm, Signal B: 254 nm;
[0456] Column Temperature: 40.degree. C.
[0457] Column: HALO C18 4.6*50 mm, 2.7 .mu.m.
TABLE-US-00001 Method Solvents Gradient Polarity A Solvent A:
H.sub.2O 0.00 min: A: 95.0% B: Positive (0.1% formic acid) 5.0%
Solvent B: MeCN 1.00 min: A: 5.0% B: (0.1% formic acid) 95.0% 2.00
min: A: 5.0% B: 95.0% 2.01 min: A: 95.0% B: 5.0% 2.50 min: A: 95.0%
B: 5.0% B Solvent A: H.sub.2O 0.00 min: A: 95.0% B: Negative (0.1%
formic acid) 5.0% Solvent B: MeCN 1.00 min: A: 5.0% B: (0.1% formic
acid) 95.0% 2.00 min: A: 5.0% B: 95.0% 2.01 min: A: 95.0% B: 5.0%
2.50 min: A: 95.0% B: 5.0% C Solvent A: H.sub.2O (0.025% 0.00 min:
A: 95.0% B: Positive trifluoroacetic acid) 5.0% Solvent B: MeCN
(0.025% 1.00 min: A: 5.0% B: trifluoroacetic acid) 95.0% 2.00 min:
A: 5.0% B: 95.0% 2.01 min: A: 95.0% B: 5.0% 2.50 min: A: 95.0% B:
5.0%
[0458] LCMS Method D/E
[0459] Analytical HPLC was conducted on a X-Terra MS C18 column
(3.5 .mu.m 30.times.4.6 mm id) eluting with 0.01M ammonium acetate
in water (solvent A) and 100% acetonitrile (solvent B), using the
following elution gradient 0-4 minutes: 5% to 100% B, 4-5 minutes
100% B, at a flow rate of 1.4 ml/minute at 40.degree. C.
[0460] The mass spectra (MS) were recorded on a Waters ZQ mass
spectrometer using electrospray positive ionisation [ES+ to give
MH.sup.+ molecular ions] or electrospray negative ionisation [ES-
to give (M-H)- molecular ions] modes. Cone voltage: 20V (method D)
or 40V (method E).
[0461] LCMS Method F/G
TABLE-US-00002 Method Description F Column: Acquity BEH C18 (50 mm
.times. 2.1 mm, 1.7 .mu.m) Mobile Phase: A: 0.1% Formic acid in
water; B: 0.1% Formic acid in MeCN Time (min)/% B: 0/3, 0.4/3,
3.2/98, 3.8/98, 4.2/3, 4.5/3 Column Temp: 35.degree. C. Flow Rate:
0.6 mL/min G Column: X Bridge C18 (50 mm .times. 4.6 mm, 2.5 .mu.m)
Mobile Phase: A: 5 mM Ammonium bicarbonate in water (pH~10); B:
MeCN Time (min)/% B: 0/5, 0.5/5, 1/15, 3.3/98, 5.2/98, 5.5/5, 6/5.
Column Temp: 35.degree. C. Flow Rate: 1.3 mL/min
Intermediate 1: 4-Chloro-5-methyl-2-nitrophenol
##STR00011##
[0463] 4-Chloro-3-methylphenol (500 g, 3.51 mol) was dissolved in
acetic acid (2.5 L) at 10.degree. C., then nitric acid (70%, 410 g,
4.56 mol) was added drop-wise and maintaining a temperature of
<30.degree. C. Once the addition was complete the mixture was
stirred at between 20 and 30.degree. C. for 1 h. After the
completion of the reaction, the reaction mixture was filtered, the
solid was carefully washed with acetic acid (200 mL) and water (500
mL), the solid was collected, and air dried to give
4-chloro-5-methyl-2-nitrophenol as a yellow solid (308 g, 47%).
[0464] .sup.1H NMR (300 MHz, CDCl.sub.3) .delta. 10.43 (s, 1H),
8.09 (s, 1H), 7.05 (s, 1H), 2.43 (s, 3H).
Intermediate 2: 2-Amino-4-chloro-5-methylphenol
##STR00012##
[0466] 4-Chloro-5-methyl-2-nitrophenol (Intermediate 1, 150 g,
0.802 mol), iron trichloride (15 g, 10% wt), active carbon (15 g,
10% wt) in methanol (600 mL) and the reaction mixture was heated to
80.degree. C. Hydrazine hydrate (80%, 502 g, 8.02 mol) was added
drop-wise and the reaction mixture was stirred at 80.degree. C. for
15 h. The reaction mixture was filtered, and the filtrated was
collected, the solvent removed and the residue was dissolved in
ethyl acetate (300 mL). The ethyl acetate was washed with water
(200 mL), then with brine (200 mL.times.3) and dried over sodium
sulphate. The solvent was removed to give
2-amino-4-chloro-5-methylphenol as a purple solid (100 g, 79%).
[0467] LCMS (A): Rt 1.32 min, MH.sup.+ 158/160.
Intermediate 3: 5-Chloro-6-methylbenzo[d]oxazol-2(3H)-one
##STR00013##
[0469] A mixture of 2-amino-4-chloro-5-methylphenol (Intermediate
2, 100 g, 0.637 mol), N,N'-carbonyldiimidazole (155 g, 0.955 mol)
in THF (500 mL) was stirred at 66.degree. C. for 1 h, the solvent
was removed and the residue was diluted with water (1 L). The solid
was isolated by filtration, dissolved in ethyl acetate (500 mL) and
dried over sodium sulphate. The mixture was filtered and the
filtrate and the solvent evaporated under reduced pressure to give
5-chloro-6-methylbenzo[d]oxazol-2(3H)-one as a yellow solid (100 g,
86%).
[0470] .sup.1H NMR (300 MHz, d.sub.6-DMSO) .delta. 11.73 (bs, 1H),
7.35 (s, 1H), 7.16 (s, 1H), 2.34 (s, 3H).
Intermediate 4: 4-Chloro-1-methoxy-2-nitrobenzene
##STR00014##
[0472] Sodium hydroxide (44.16 g, 1.104 mol) was dissolved in
methanol (500 mL) and the methanol solution was added to
1,4-dichloro-2-nitrobenzene (100 g, 0.521 mol) in methanol at
70.degree. C. After the addition the reaction mixture was stirred
at 70.degree. C. for 2 h, then was cooled and poured into water (3
L). The solid was isolated by filtration and washed with water (2
L) to give 4-chloro-1-methoxy-2-nitrobenzene as a pale-yellow solid
(93 g, 95%).
[0473] .sup.1H NMR (300 MHz, CDCl.sub.3) .delta. 7.83 (d, J=2.4 Hz,
1H), 7.50 (dd, J=8.7 Hz, J=2.4 Hz, 1H), 7.03 (d, J=8.7 Hz, 1H),
3.95 (s, 3H).
Intermediate 5: 5-Chloro-2-methoxyaniline
##STR00015##
[0475] Hydrazine hydrate (80%, 186 g, 2.975 mol) was added
drop-wise to a refluxing mixture of
4-chloro-1-methoxy-2-nitrobenzene (Intermediate 4, 93 g, 0.496
mol), iron trichloride (9.3 g, 10% wt) and active carbon (9.3 g,
10% wt) in methanol (1 L). After the addition, the reaction mixture
was stirred at reflux for 16 h, the reaction mixture was filtered
and the solvent evaporated from the filtrate under vacuum. The
residue was washed with petroleum ether (2 L) to give
5-chloro-2-methoxyaniline as a white solid (76.7 g, 98%).
[0476] .sup.1H NMR (300 MHz, CDCl.sub.3) .delta. 6.67-6.65 (m, 3H),
3.83 (bs, 2H), 3.81 (s, 3H).
Intermediate 6: 4-Bromo-5-chloro-2-methoxyaniline
##STR00016##
[0478] A mixture of 5-chloro-2-methoxyaniline (Intermediate 5, 76.7
g, 0.487 mol) in acetonitrile (500 mL) at 0.degree. C. was treated
with N-bromosuccinimide (86.7 g, 0.487 mol) added in portions over
2 h. After the addition, the solvent was removed and the residue
purified by column chromatography (silica: 200-300 mesh, 200 g)
eluting with petroleum ether/ethyl acetate from 20:1 to 4:1) to
give 4-bromo-5-chloro-2-methoxyaniline as a light-yellow solid
(40.2 g, 35%).
[0479] LCMS(A): Rt 1.63 min, MH.sup.+ 236/238.
[0480] .sup.1H NMR (300 MHz, CDCl.sub.3) .delta. 6.93 (s, 1H), 6.78
(s, 1H), 3.83 (s, 3H).
Intermediate 7: 2-Amino-5-bromo-4-chlorophenol
##STR00017##
[0482] 4-Bromo-5-chloro-2-methoxyaniline (Intermediate 6, 10 g,
42.3 mmol) in DCM (200 mL) was cooled to 0.degree. C. Boron
tribromide (21.2 g, 84.6 mmol) was added drop-wise over 30 min, the
reaction mixture was stirred at 0.degree. C. for 1 h and then at
room temperature for 14 h. The reaction mixture was poured into ice
water, sodium hydrogen carbonate was added until pH>7 and
aqueous layer was extracted with ethyl acetate (300 mL.times.3).
The organic phase was dried over magnesium sulphate and the solvent
removed under vacuum to give 2-amino-5-bromo-4-chlorophenol as a
red solid (8.4 g, 89%, used without further purification for the
next step).
[0483] LCMS (A): Rt 1.46 min, MH.sup.+ 222/224.
Intermediate 8: 6-Bromo-5-chlorobenzo[d]oxazol-2(3H)-one
##STR00018##
[0485] A mixture of 2-amino-5-bromo-4-chlorophenol (Intermediate 7,
8.4 g, 37.8 mmol), N,N-carbonyldiimidazole (12.2 g, 75.6 mmol) in
THF (250 mL), was heated at 80.degree. C. for 3 h. The solvent was
removed under vacuum and the residue purified by column
chromatography (silica: 200-400 mesh, 100 g) eluting with petroleum
ether/ethyl acetate from 9:1 to 5:1) to give
6-bromo-5-chlorobenzo[d]oxazol-2(3H)-one as an orange solid (7.7 g,
82%).
[0486] LCMS (A): Rt 1.49 min, MH.sup.+ 248/250.
Intermediate 9: Methyl
3-(6-bromo-5-chloro-2-oxobenzo[d]oxazol-3(2H)-yl)propanoate
##STR00019##
[0488] A mixture of 6-bromo-5-chlorobenzo[d]oxazol-2(3H)-one
(Intermediate 8, 7.7 g, 31 mmol), methyl 3-bromopropanoate (6.2 g,
37 mmol), potassium carbonate (8.5 g, 62 mmol) in acetonitrile (200
mL), was stirred at 70.degree. C. for 16 h, the solvent was
evaporated under vacuum, the residue purified by column
chromatography (silica: 200-400 mesh, 50 g) eluting with petroleum
ether/ethyl acetate 4:1 to give methyl
3-(6-bromo-5-chloro-2-oxobenzo[d]oxazol-3(2H)-yl)propanoate as an
orange solid (6.2 g, 62%).
[0489] LCMS (A): Rt 1.66 min, MH.sup.+=334/336.
Intermediate 10: Methyl 3-(5-chloro-2-oxo-6-(4, 4, 5,
5-tetramethyl-1, 3,
2-dioxaborolan-2-yl)benzo[d]oxazol-3(2H)-yl)propanoate
##STR00020##
[0491] A mixture of methyl
3-(6-bromo-5-chloro-2-oxobenzo[d]oxazol-3(2H)-yl)propanoate
(Intermediate 9, 5.2 g, 15.56 mmol), bis(pinacolato)diboron (11.8
g, 46.7 mmol), [1, 1'-Bis(diphenylphosphino)
ferrocene]dichloropalladium (II) (568 mg, 0.778 mmol), potassium
acetate (3 g, 31.12 mmol) in 1, 4-dioxane (500 mL) was stirred at
100.degree. C. for 16 h under a nitrogen atmosphere. The solvent
was evaporated under vacuum and the residue purified by column
chromatography (silica: 200-300 mesh, 80 g) eluting with petroleum
ether/ethyl acetate=9:1 then 8:1) to give methyl
3-(5-chloro-2-oxo-6-(4, 4, 5, 5-tetramethyl-1, 3,
2-dioxaborolan-2-yl)benzo[d]oxazol-3(2H)-yl)propanoate as a yellow
oil (5.2 g, 87%).
[0492] LCMS (A): Rt 1.73 min, MH.sup.+ 382/384.
Intermediate 11: Methyl
3-(5-chloro-6-hydroxy-2-oxobenzo[d]oxazol-3(2H)-yl)propanoate
##STR00021##
[0494] A mixture of methyl 3-(5-chloro-2-oxo-6-(4, 4, 5,
5-tetramethyl-1, 3,
2-dioxaborolan-2-yl)benzo[d]oxazol-3(2H)-yl)propanoate
(Intermediate 10, 5.2 g, 13.6 mmol) in hydrogen peroxide (30%, 12
mL), acetic acid (10 mL) and THF (100 mL) was stirred at room
temperature for 16 h. Water was added, the mixture extracted with
ethyl acetate (100 mL.times.3), the organic phase was dried over
magnesium sulphate and the solvent evaporated under vacuum. The
residue was purified by column chromatography (silica: 200-300
mesh, 50 g) eluting with petroleum ether/ethyl acetate=9:1, 8:1,
6:1, 4:1) to give methyl
3-(5-chloro-6-hydroxy-2-oxobenzo[d]oxazol-3(2H)-yl)propanoate as a
yellow solid (2.5 g, 68%).
[0495] LCMS (A): Rt 1.32 min, MH.sup.+ 272.
Intermediate 12: Methyl
3-(5-chloro-2-oxo-6-(1-(pyridin-2-yl)ethoxy)benzo[d]oxazol-3(2H)-yl)propa-
noate
##STR00022##
[0497] 1-(Pyridin-2-yl)ethanol (71 mg, 0.57 mmol), diethyl
azodicarboxylate (84 mg, 0.48 mmol) and triphenyphosphine (126 mg,
0.48 mmol) were added to methyl
3-(5-chloro-6-hydroxy-2-oxobenzo[d]oxazol-3(2H)-yl)propanoate
(Intermediate 11, 0.13 g, 0.48 mmol) in toluene (10 mL) and the
reaction was stirred at room temperature for 2 days. The reaction
mixture was poured into water (15 mL), aqueous layer was extracted
with ethyl acetate (10 mL.times.3), combined organic layer and
dried over sodium sulphate and partially purified by column
chromatography [silica: 200-300 mesh, 10 g] eluting with petroleum
ether/ethyl acetate=4:1] to give methyl
3-(5-chloro-2-oxo-6-(1-(pyridin-2-yl)ethoxy)benzo[d]oxazol-3(2H)-yl)propa-
noate as a white solid (270 mg, crude).
[0498] LCMS (A): Rt 1.57 min, MH.sup.+ 377.
Intermediate 13: (R)-Methyl
3-(5-chloro-2-oxo-6-(1-(pyridin-2-yl)ethoxy)benzo[d]oxazol-3(2H)-yl)propa-
noate
##STR00023##
[0500] To methyl
3-(5-chloro-6-hydroxy-2-oxobenzo[d]oxazol-3(2H)-yl)propanoate
(Intermediate 11, 133 mg, 0.49 mmol) in toluene (10 mL), was added
(S)-1-(pyridin-2-yl)ethanol (60 mg, 0.49 mmol), triphenylphosphine
(93 mg, 0.74 mmol), diethyl azodicarboxylate (129 mg, 0.74 mmol)
and the reaction mixture was stirred at room temperature for 16 h.
The solvent was removed and the residue was purified by column
chromatography (silica: 200-300 mesh, 5 g) eluting with petroleum
ether/ethyl acetate 5:1 to give (R)-methyl
3-(5-chloro-2-oxo-6-(1-(pyridin-2-yl)ethoxy)benzo[d]oxazol-3(2H)-yl)propa-
noate as a colourless oil (170 mg, 90%). LCMS (A): Rt 1.40 min,
MH.sup.+=377/379.
Intermediate 14: (S)-methyl
3-(5-chloro-2-oxo-6-(1-(pyridin-2-yl)ethoxy)benzo[d]oxazol-3(2H)-yl)propa-
noate
##STR00024##
[0502] To methyl
3-(5-chloro-6-hydroxy-2-oxobenzo[d]oxazol-3(2H)-yl)propanoate
(Intermediate 11, 133 mg, 0.49 mmol) in toluene (10 mL), was added
(R)-1-(pyridin-2-yl)ethanol (60 mg, 0.49 mmol), triphenylphosphine
(193 mg, 0.74 mmol), diethyl azodicarboxylate (129 mg, 0.74 mmol),
the reaction mixture was stirred at room temperature for 16 h. The
solvent was removed and the residue was purified by column
chromatography (silica: 200-300 mesh, 5 g) eluting with petroleum
ether/ethyl acetate 5:1) to give (S)-methyl
3-(5-chloro-2-oxo-6-(1-(pyridin-2-yl)ethoxy)benzo[d]oxazol-3(2H)-yl)propa-
noate as a colourless oil (160 mg, 86%). LCMS (A): Rt 1.60 min,
MH.sup.+=377/379.
Example 1
3-(5-Chloro-6-methyl-2-oxobenzo[d]oxazol-3(2H)-yl)propanoic
acid
##STR00025##
[0504] 5-Chloro-6-methylbenzo[d]oxazol-2(3H)-one (Intermediate 3,
150 g, 0.82 mol), 3-bromopropanoic acid (251 g, 1.64 mol),
potassium carbonate (226 g, 1.64 mol) were mixed in acetonitrile (2
L) and the reaction mixture stirred at 80.degree. C. for 18 h. The
reaction mixture was cooled to ambient temperature and filtered.
The solid was collected and acidified to pH 4-5 with hydrochloric
acid (3 N) and extracted with ethyl acetate (300 mL.times.5). The
combined organic phases were dried over sodium sulphate, filtered,
the filtrate was collected and the solvent evaporated to give
3-(5-chloro-6-methyl-2-oxobenzo[d]oxazol-3(2H)-yl)propanoic acid as
a yellow solid (220 g). Used without further purification.
[0505] .sup.1H NMR (300 MHz, d.sub.6-DMSO) .delta. 12.43 (bs, 1H),
7.51 (s, 1H), 7.36 (s, 1H), 4.00 (t, J=6 Hz, 2H), 2.70 (t, J=6 Hz,
2H), 2.33 (s, 3H).
Example 1 (Alternative Preparation)
3-(5-Chloro-6-methyl-2-oxobenzo[d]oxazol-3(2H)-yl)propanoic
acid
##STR00026##
[0507] A mixture of 5-chloro-6-methylbenzo[d]oxazol-2(3H)-one (250
mg, 1.362 mmol, commercial), potassium carbonate (565 mg, 4.09
mmol) and 3-bromopropanoic acid (417 mg, 2.72 mmol) in acetonitrile
(10 mL) was heated to 90.degree. C. and stirred for 2 h. The
mixture was cooled and filtered; the isolated white powder was
washed with acetonitrile, suspended in water and acidified to pH 2
with hydrochloric acid (1N). The resulting solid was then
triturated with hot acetonitrile, filtered and dried to give the
3-(5-chloro-6-methyl-2-oxobenzo[d]oxazol-3(2H)-yl)propanoic acid as
a white powder (225 mg, 64.6%).
[0508] LCMS (E): Rt 2.29 min, [M-H].sup.- 254/256
Example 1a Tris Salt Formation
3-(5-chloro-6-methyl-2-oxobenzo[d]oxazol-3(2H)-yl)propanoic acid,
2-Amino-2-(hydroxymethyl)-1,3-propanediol salt
##STR00027##
[0510] A mixture of
3-(5-chloro-6-methyl-2-oxobenzo[d]oxazol-3(2H)-yl) propanoic acid
(Example 1, 220 g, 0.863 mol),
2-amino-2-(hydroxymethyl)propane-1,3-diol (104.5 g, 0.863 mol) in
ethanol (4 L), was stirred at 80.degree. C. for 2 h, the reaction
mixture was filtered, the filtrate was collected and cooled to
ambient temperature. The mixture was filtered, the solid was washed
with ethanol (200 mL) and dried over air to give
3-(5-chloro-6-methyl-2-oxobenzo[d]oxazol-3(2H)-yl)propanoic acid,
2-amino-2-(hydroxymethyl)-1,3-propanediol salt as an off-white
solid (150 g, 46%).
[0511] LCMS (A): Rt 1.41 min, MH.sup.+ 256/258.
[0512] .sup.1H NMR (300 MHz, CD.sub.3OD) .delta. 7.37 (s, 1H), 7.16
(s, 1H), 4.05 (t, J=6 Hz, 2H), 3.64 (s, 6H), 2.59 (t, J=6 Hz, 2H),
2.37 (s, 3H).
Example 2
3-(5-chloro-2-oxo-6-(1-(pyridin-2-yl)ethoxy)benzo[d]oxazol-3(2H)-yl)propan-
oic acid, 2-Amino-2-(hydroxymethyl)-1, 3-propanediol salt
##STR00028##
[0514] Methyl
3-(5-chloro-2-oxo-6-(1-(pyridin-2-yl)ethoxy)benzo[d]oxazol-3(2H)-yl)propa-
noate (Intermediate 12, 270 mg) in hydrochloric acid (0.5 N, 5 mL)
was added to 1,4-dioxane (5 mL) and the reaction mixture was
stirred at 80.degree. C. for 3 h. The solvent was removed, the
residue was purified by prep-HPLC [eluent: MeCN-water (0.1% TFA),
gradient: 60-90% MeCN] to give
3-(5-chloro-2-oxo-6-(1-(pyridin-2-yl)ethoxy)benzo[d]oxazol-3(2H)-yl)-
propanoic acid as a yellow oil (30 mg, 0.08 mmol). Methanol (2 mL)
and 2-amino-2-(hydroxymethyl)-1, 3-propanediol (10 mg, 0.08 mmol)
were added and the reaction mixture was stirred at room temperature
for 30 minutes. The mixture was concentrated to give
3-(5-chloro-2-oxo-6-(1-(pyridin-2-yl)ethoxy)benzo[d]oxazol-3(2H)-yl)propa-
noic acid, 2-Amino-2-(hydroxymethyl)-1, 3-propanediol salt as a
yellow oil (40 mg, 17%).
[0515] LCMS (A): Rt 1.36 min, MH.sup.+ 363.
[0516] .sup.1H NMR (300 MHz, CD.sub.3OD) .delta. 8.52 (d, J=6 Hz,
1H), 7.82 (t, J=6 Hz, 1H), 7.57 (d, J=9 Hz, 1H), 7.41 (s, 1H),
7.35-7.31 (m, 1H), 6.91 (s, 1H), 5.44 (dd, J=12, 6 Hz, 2H), 4.02
(t, J=6 Hz, 2H), 3.66 (s, 6H), 2.62 (t, J=6 Hz, 2H), 1.69 (d, J=6
Hz, 3H).
Example 3
(R)-3-(5-chloro-2-oxo-6-(1-(pyridin-2-yl)ethoxy)benzo[d]oxazol-3(2H)-yl)pr-
opanoic acid
##STR00029##
[0518] Hydrochloric acid (0.5 N, 4 mL) was added to a solution of
(R)-methyl
3-(5-chloro-2-oxo-6-(1-(pyridin-2-yl)ethoxy)benzo[d]oxazol-3(2H)-yl)propa-
noate (Intermediate 13, 170 mg) in 1,4-dioxane (4 mL) and the
reaction mixture was stirred at 80.degree. C. for 16 h. The solvent
was removed and the residue was purified by prep-HPLC
(acetonitrile/water (0.1% TFA) 20:80 to 70:30). The solvent was
removed and the residue was further purified by Super Critical
Fluid Chromatography (hexane/ethanol 1:1, 0.2% TFA) to give
(R)-3-(5-chloro-2-oxo-6-(1-(pyridin-2-yl)ethoxy)benzo[d]oxazol-3(2H)-yl)p-
ropanoic acid as a yellow oil (30 mg, 18%). LCMS (A): Rt 1.37 min,
MH.sup.+=363/365.
Example 3a (Tris Salt Formation)
(R)-3-(5-chloro-2-oxo-6-(1-(pyridin-2-yl)ethoxy)benzo[d]oxazol-3(2H)-yl)pr-
opanoic acid, 2-Amino-2-(hydroxymethyl)-1,3-propanediol salt
##STR00030##
[0520] 2-Amino-2-(hydroxymethyl)propane-1,3-diol (10 mg, 0.08 mol)
was added to
(R)-3-(5-chloro-2-oxo-6-(1-(pyridin-2-yl)ethoxy)benzo[d]oxazol-3-
(2H)-yl)propanoic acid (Example 3 (free acid), 30 mg, 0.08 mmol) in
methanol (2 mL), the reaction mixture was stirred at ambient
temperature for 30 min and the solvent removed under vacuum to give
(R)-3-(5-chloro-2-oxo-6-(1-(pyridin-2-yl)ethoxy)benzo[d]oxazol-3(2H)-yl)p-
ropanoic acid, 2-Amino-2-(hydroxymethyl)-1,3-propanediol salt as a
yellow oil (40 mg, 100%).
[0521] LCMS (A): Rt 1.38 min, MH.sup.+=363/365.
[0522] .sup.1H NMR (300 MHz, CD.sub.3OD) .delta. 8.51 (d, J=3 Hz,
1H), 7.82 (td, J=9, 3 Hz, 1H), 7.57 (d, J=9 Hz, 1H), 7.42 (s, 1H),
7.34-7.33 (m, 1H), 6.91 (s, 1H), 5.43 (q, J=6 Hz, 1H), 4.02 (t, J=6
Hz, 2H), 3.66 (s, 6H), 2.59 (t, J=6 Hz, 2H), 1.69 (d, J=6 Hz, 3H).
Chiral-HPLC: 214 nm (100.0%), 254 nm (100.0%).
Example 4
(S)-3-(5-chloro-2-oxo-6-(1-(pyridin-2-yl)ethoxy)benzo[d]oxazol-3(2H)-yl)pr-
opanoic acid
##STR00031##
[0524] Hydrochloric acid (0.5 N, 4 mL) was added to (S)-methyl
3-(5-chloro-2-oxo-6-(1-(pyridin-2-yl)ethoxy)benzo[d]oxazol-3(2H)-yl)propa-
noate (Intermediate 14, 160 mg) in 1,4-dioxane (4 mL) and the
reaction mixture was stirred at 80.degree. C. for 16 h. The solvent
was removed, the residue was purified with prep-HPLC
(acetonitrile/water (0.1% TFA) 20:80 to 70:30). The solvent was
removed, and the residue was further purified by Super Critical
Fluid Chromatography (hexane/Ethanol 1:1, 0.2% TFA) to give
(S)-3-(5-chloro-2-oxo-6-(1-(pyridin-2-yl)ethoxy)benzo[d]oxazol-3(2H)-yl)p-
ropanoic acid (23 mg, 5%).
[0525] LCMS (A): Rt 1.37 min, MH.sup.+=363/365.
[0526] Chiral-HPLC: 214 nm (100.0%), 254 nm (100.0%).
[0527] Sodium hydroxide (44.16 g, 1.104 mol) was dissolved in
methanol (500 mL) and the methanol solutions was added to 1,
4-dichloro-2-nitrobenzene (100 g, 0.521 mol) in methanol at
70.degree. C. After the addition the reaction mixture was stirred
at 70.degree. C. for 2 h, then was cooled and poured into water (3
L). The solid was isolated by filtration and washed with water (2
L) to give 4-chloro-1-methoxy-2-nitrobenzene as a pale-yellow solid
(93 g, 95%).
Example 4a (Tris Salt Formation)
(S)-3-(5-chloro-2-oxo-6-(1-(pyridin-2-yl)ethoxy)benzo[d]oxazol-3(2H)-yl)pr-
opanoic acid, 2-amino-2-(hydroxymethyl)-1,3-propanediol salt
##STR00032##
[0529] 2-Amino-2-(hydroxymethyl)propane-1,3-diol (8 mg, 0.063 mol)
was added to
(S)-3-(5-chloro-2-oxo-6-(1-(pyridin-2-yl)ethoxy)benzo[d]oxazol-3-
(2H)-yl)propanoic acid (Example 4 (free acid), 23 mg, 0.063 mmol),
in methanol (2 mL) and the reaction mixture was stirred at room
temperature for 30 min. The solvent was removed and the residue was
purified by prep-HPLC (acetonitrile/water (0.1% TFA) 20:80 to
70:30). After solvent removal, methanol (2 mL) and
2-amino-2-(hydroxymethyl)propane-1,3-diol (3 mg, 0.024 mmol) were
added, the reaction mixture was stirred at room temperature for 30
min. Removal of solvent gave
(S)-3-(5-chloro-2-oxo-6-(1-(pyridin-2-yl)ethoxy)benzo[d]oxazol-3(2H)-yl)p-
ropanoic acid, 2-amino-2-(hydroxymethyl)-1,3-propanediol salt as a
brown solid (10 mg, 13%).
[0530] LCMS (A): Rt 1.37 min, MH.sup.+=363/365.
[0531] .sup.1H NMR (300 MHz, CD.sub.3OD) .delta. 8.51 (d, J=3 Hz,
1H), 7.82 (td, J=9, 3 Hz, 1H), 7.57 (d, J=9 Hz, 1H), 7.42 (s, 1H),
7.34-7.33 (m, 1H), 6.91 (s, 1H), 5.43 (q, J=6 Hz, 1H), 4.02 (t, J=6
Hz, 2H), 3.66 (s, 6H), 2.59 (t, J=6 Hz, 2H), 1.69 (d, J=6 Hz, 3H).
Chiral-HPLC: 214 nm (100.0%), 254 nm (100.0%).
[0532] .sup.1H NMR (300 MHz, CDCl.sub.3) .delta. 7.83 (d, J=2.4 Hz,
1H), 7.50 (dd, J=8.7 Hz, J=2.4 Hz, 1H), 7.03 (d, J=8.7 Hz, 1H),
3.95 (s, 3H).
[0533] Examples 5, 6, 8, 9, 11, and 25 were prepared in a manner
substantially according to Scheme 1 or using methyl
3-(6-bromo-5-chloro-2-oxobenzo[d]oxazol-3(2H)-yl)prapanoate (II).
Examples 7, 10, 12-24 and 26-37 were prepared in a manner
substantially similar to Example 2 according to Scheme 2.
TABLE-US-00003 TABLE 1 Example Molecular Rt LCMS no. Structure Name
ion (min) method 5 ##STR00033## 3-(5,6- dichloro-2- oxobenzo[d]ox-
azol-3(2H)- yl)propanoic acid MH+ 276 2.02 D 6 ##STR00034##
3-(5-chloro-6- fluoro-2- oxobenzo[d]ox- azol-3(2H)- yl)propanoic
acid MH+ 260 1.85 D 7 ##STR00035## 3-(5-chloro-6- methoxy-2-
oxobenzo[d]ox- azol-3(2H)- yl)propanoc acid MH+ 272 1.37 C 8
##STR00036## 3-(5-chloro-6- cyano-2- oxobenzo[d]ox- azol-3(2H)-
yl)propanoic acid [M - H]- 265 1.38 B 9 ##STR00037## 2-amino-2-
(hydroxymethyl) propane-1,3- diol 3-(6- bromo-5- chloro-2-
oxobenzo[d]ox- azol-3(2H)- yl)propanoate [M - H]- 318 1.46 B 10
##STR00038## 2-amino-2- (hydroxymethyl) propane-1,3- diol 3-(5-
chloro-6- ethoxy-2- oxobenzo[d]ox- azol-3(2H)- yl)propanoate [M -
H]- 284 1.43 B 11 ##STR00039## 2-amino-2- (hydroxymethyl)
propane-1,3- diol 3-(5- chloro-6-ethyl- 2- oxobenzo[d]ox-
azol-3(2H)- yl)propanoate [M - H]- 268 1.55 B 12 ##STR00040##
2-amino-2- (hydroxymethyl) propane-1,3- diol 3-(5- chloro-6-
isobutoxy-2- oxobenzo[d]ox- azol-3(2H)- yl)propanoate MH+ 314 2.48
E 13 ##STR00041## 2-amino-2- (hydroxymethyl) propane-1,3- diol
3-(5- chloro-6- (cyclopropylmeth- oxy)-2- oxobenzo[d]ox-
azol-3(2H)- yl)propanoate MH+ 312 1.51 A 14 ##STR00042## 2-amino-2-
(hydroxymethyl) propane-1,3- diol 3-(5- chloro-6- (cylcobutylmeth-
oxy)-2- oxobenzo[d]ox- azol-3(2H)- yl)propanoate MH+ 326 1.57 A 15
##STR00043## 2-amino-2- (hydroxymethyl) propane-1,3- diol 3-(6-
(benzyloxy)-5- chloro-2- oxobenzo[d]ox- azol-3(2H)- yl)propanoate
MH+ 348 1.55 A 16 ##STR00044## 2-amino-2- (hydroxymethyl)
propane-1,3- diol 3-(5- chloro-6-(2- methoxyethoxy)- 2-
oxobenzo[d]ox- azol-3(2H)- yl)propanoate MH+ 316 1.40 A 17
##STR00045## 2-amino-2- (hydroxymethyl) propane-1,3- diol 3-(5-
chloro-2-oxo- 6- propoxybenzo[d] oxazol- 3(2H)- yl)propanoate MH+
300 2.29 E 18 ##STR00046## 2-amino-2- (hydroxymethyl) propane-1,3-
diol 3-(5- chloro-2-oxo- 6-(3,3,3- trifluoropropoxy) benzo[d]oxa-
zol-3(2H)- yl)propanoate MH+ 354 1.47 A 19 ##STR00047## 2-amino-2-
(hydroxymethyl) propane-1,3- diol 3-(5- chloro-2-oxo- 6-(pyridin-2-
ylmethoxy)ben- zo[d]oxazol- 3(2H)- yl)propanoate MH+ 349 1.34 A 20
##STR00048## 2-amino-2- (hydroxymethyl) propane-1,3- diol 3-(5-
chloro-6- isopropoxy-2- oxobenzo[d]ox- azol-3(2H)- yl)propanoate
MH+ 300 2.23 E 21 ##STR00049## 2-amino-2- (hydroxymethyl)
propane-1,3- diol 3-(5- chloro-6- cyclobutoxy-2- oxobenzo[d]ox-
azol-3(2H)- yl)propanoate [M - H]- 310 1.56 B 22 ##STR00050##
2-amino-2- (hydroxymethyl) propane-1,3- diol 3-(5- chloro-6-
cyclopropoxy- 2- oxobenzo[d]ox- azol-3(2H)- yl)propanoate [M - H]-
296 1.51 B 23 ##STR00051## 2-amino-2- (hydroxymethyl) propane-1,3-
diol 3-(5- chloro-2-oxo- 6-(2,2,2- trifluoroethoxy) benzo[d]oxazol-
3(2H)- yl)propanoate MH+ 340 1.48 A 24 ##STR00052## 2-amino-2-
(hydroxymethyl) propane-1,3- diol 3-(5- chloro-6- (oxetan-3-
yloxy)-2- oxobenzo[d]ox- azol-3(2H)- yl)propanoate [M - H]- 312
1.28 B 25 ##STR00053## 2-amino-2- (hydroxymethyl) propane-1,3- diol
3-(5- chloro-6- isbotuyl-2- oxobenzo[d]ox- azol-3(2H)-
yl)propanoate [M - H]- 296 1.64 B 26 ##STR00054## 2-amino-2-
(hydroxymethyl) propane-1,3- diol 3-(5- chloro-6- (oxetan-3-
ylmethoxy)-2- oxobenzo[d]ox- azol-3(2H)- yl)propanoate [M - H]- 326
1.36 B 27 ##STR00055## 2-amino-2- (hydroxymethyl) propane-1,3- diol
3-(5- chloro-6-((1- methoxypropan- 2-yl)oxy)-2- oxobenzo[d]ox-
azol-3(2H)- yl)propanoate MH+ 330 1.42 A 28 ##STR00056## 2-amino-2-
(hydroxymethyl) propane-1,3- diol 3-(5- chloro-6-((6-
methylpyridin- 3-yl)methoxy)- 2- oxobenzo[d]ox- azol-3(2H)-
yl)propanoate MH+ 363 1.21 A 29 ##STR00057## 2-amino-2-
(hydroxymethyl) propane-1,3- diol 3-(5- chloro-6-((2-
methylpyridin- 4-yl)methoxy)- 2- oxobenzo[d]ox- azol-3(2H)-
yl)propanoate MH+ 363 1.18 A 30 ##STR00058## 2-(((3-(2-
carboxyethyl)- 5-chloro-2- oxo-2,3- dihydrobenzo[d] oxazol-6-
yl)oxy)methyl) pyridine 1- oxide MH+ 365 1.30 A 31 ##STR00059##
2-amino-2- (hydroxymethyl) propane-1,3- diol 3-(5- chloro-6-((1-
methyl-1H- imidazol-2- yl)methoxy)-2- oxobenzo[d]ox- azol-3(2H)-
yl)propanoate MH+ 352 1.01 A 32 ##STR00060## 2-amino-2-
(hydroxymethyl) propane-1,3- diol 3-(5- chloro-6-((2-
methylpyridin- 3-yl)methoxy)- 2- oxobenzo[d]ox- azol-3(2H)-
yl)propanoate MH+ 363 1.23 A 33 ##STR00061## 2-amino-2-
(hydroxymethyl) propane-1,3- diol 3-(5- chloro-2-oxo-
6-(pyrimidin-2- ylmethoxy)ben- zo[d]oxazol- 3(2H)- yl)propanoate
MH+ 350 1.34 A 34 ##STR00062## 2-amino-2- (hydroxymethyl)
propane-1,3- diol 3-(5- chloro-6-((2- methyloxazol- 5-yl)methoxy)-
2- oxobenzo[d]ox- azol-3(2H)- yl)propanoate MH+ 353 1.35 A 35
##STR00063## 2-amino-2- (hydroxymethyl) propane-1,3- diol 3-(5-
chloro-6-((5- methyloxazol- 2-yl)methoxy)- 2- oxobenzo[d]ox-
azol-3(2H)- yl)propanoate MH+ 353 1.46 A 36 ##STR00064## 3-(6-((1H-
imidazol-2- yl)methoxy)-5- chloro-2- oxobenzo[d]ox- azol-3(2H)-
yl)propanoic acid MH+ 338 1.13 A 37 ##STR00065## 2-amino-2-
(hydroxymethyl) propane-1,3- diol 3-(5- chloro-2-oxo-
6-(pyridazin-3- ylmethoxy)ben- zo[d]oxazol- 3(2H)- yl)propanoate
MH+ 350 1.50 A
[0534] Examples 3b-3l and 38-61 were prepared in a manner
substantially according to Scheme 1 or using methyl
3-(6-bromo-5-chloro-2-oxobenzo[d]oxazol-3(2H)-yl)prapanoate
(II).
TABLE-US-00004 Example Molecular Rt LCMS no. Structure Name ion
(min) Method 3b ##STR00066## (R)-3-(5- chloro-2-oxo- 6-(1-(pyridin-
2- yl)ethoxy)benz- o[d]oxazol- 3(2H)- yl)propanoic acid
hydrochloride MH+ 363 1.37 A 3c ##STR00067## (R)-3-(5-
chloro-2-oxo- 6-(1-(pyridin- 2- yl)ethoxy)benz- o[d]oxazol- 3(2H)-
yl)propanoic acid compound with sulfuric acid (1:1) MH+ 363 1.38 A
3d ##STR00068## (R)-3-(5- chloro-2-oxo- 6-(1-(pyridin- 2-
yl)ethoxy)benz- o[d]oxazol- 3(2H)- yl)propanoic acid compound with
methanesulfo- nic acid (1:1) MH+ 363 1.38 A 3e ##STR00069##
N-benzyl-2- phenylethana- mine (R)-3-(5- chloro-2-oxo-
6-(1-(pyridin- 2- yl)ethoxy)benz- o[d]oxazol- 3(2H)- yl)propanoate
MH+ 363 1.38 A 3f ##STR00070## N1,N2- dibenzylethane- 1,2-diamine
(R)-3-(5- chloro-2-oxo- 6-(1-(pyridin- 2- yl)ethoxy)benz-
o[d]oxazol- 3(2H)- yl)propanoate MH+ 363 1.37 A 3g ##STR00071##
(2R,3R,4S,5R)- 3,4,5,6- tetrahydroxy- 2- (methylamino) hexanal
(R)-3- (5-chloro-2- oxo-6-(1- (pyridin-2- yl)ethoxy)benz-
o[d]oxazol- 3(2H)- yl)propanoate MH+ 363 1.45 A 3h ##STR00072##
(R)-3-(5- chloro-2-oxo- 6-(1-(pyridin- 2- yl)ethoxy)benz-
o[d]oxazol- 3(2H)- yl)propanoic acid compound with (S)-2- amino-5-
guanidinopent- anoic acid (1:1) MH+ 363 1.38 A 3i ##STR00073##
(R)-3-(5- chloro-2-oxo- 6-(1-(pyridin- 2- yl)ethoxy)benz-
o[d]oxazol- 3(2H)- yl)propanoic acid compound with (S)-2,6-
diaminohexan- oic acid (1:1) MH+ 363 1.38 A 3j ##STR00074## sodium
(R)-3- (5-chloro-2- oxo-6-(1- (pyridin-2- yl)ethoxy)benz-
o[d]oxazol- 3(2H)- yl)propanoate MH+ 363 1.37 A 3k ##STR00075##
(R)-3-(5- chloro-2-oxo- 6-(1-(pyridin- 2- yl)ethoxy)benz-
o[d]oxazol- 3(2H)- yl)propanoic acid compound with 4-
methylbenzene- sulfonic acid (1:1) MH+ 363 1.36 A 3l ##STR00076##
N1-(2- aminoethyl)eth- ane-1,2- diamine (R)-3- (5-chloro-2-
oxo-6-(1- (pyridin-2- yl)ethoxy)benz- o[d]oxazol- 3(2H)-
yl)propanoate MH+ 363 1.38 A 38 ##STR00077## 2-amino-2-
(hydroxymethyl) propane-1,3- diol 3-(5- chloro-6-(1-(2-
methyloxazol- 5-yl)ethoxy)-2- oxobenzo[d]ox- azol-3(2H)-
yl)propanoate (racemic) MH+ 367 1.44 A 39 ##STR00078## 2-amino-2-
(hydroxymethyl) propane-1,3- diol (R)-3-(5- chloro-6-(1-(5-
fluoropyridin- 2-yl)ethoxy)-2- oxobenzo[d]ox- azol-3(2H)-
yl)propanoate MH+ 381 1.45 A 40 ##STR00079## 2-amino-2-
(hydroxymethyl) propane-1,3- diol (R)-3-(5- chloro-6-(1-(5-
methylpyridin- 2-yl)ethoxy)-2- oxobenzo[d]ox- azol-3(2H)-
yl)propanoate MH+ 377 1.49 A 40a ##STR00080## (R)-3-(5-
chloro-6-(1-(5- methylpyridin- 2-yl)ethoxy)-2- oxobenzo[d]ox-
azol-3(2H)- yl)propanoic acid MH+ 377 1.40 A 41 ##STR00081##
2-amino-2- (hydroxymethyl) propane-1,3- diol 3-(5- chloro-6-((5-
fluoropyridin- 2-yl)methoxy)- 2- oxobenzo[d]ox- azol-3(2H)-
yl)propanoate MH+ 367 1.50 A 42 ##STR00082## 2-amino-2-
(hydroxymethyl) propane-1,3- diol 3-(5- chloro-6-((5-
chloropyridin- 2-yl)methoxy)- 2- oxobenzo[d]ox- azol-3(2H)-
yl)propanoate MH+ 383 1.49 A 43 ##STR00083## 2-amino-2-
(hydroxymethyl) propane-1,3- diol (R)-3-(5- chloro-6-(1-(5-
chloropyridin- 2-yl)ethoxy)-2- oxobenzo[d]ox- azol-3(2H)-
yl)propanoate MH+ 397 1.54 A 43a ##STR00084## (R)-3-(5-
chloro-6-(1-(5- chloropyridin- 2-yl)ethoxy)-2- oxobenzo[d]ox-
azol-3(2H)- yl)propanoic acid MH+ 397 1.58 A 44 ##STR00085##
2-amino-2- (hydroxymethyl) propane-1,3- diol 3-(5- chloro-6-((5-
methylpyridin- 2-yl)methoxy)- 2- oxobenzo[d]ox- azol-3(2H)-
yl)propanoate MH+ 363 1.35 A 45 ##STR00086## 2-amino-2-
(hydroxymethyl) propane-1,3- diol (R)-3-(5- chloro-6-(1-(6-
methylpyridin- 2-yl)ethoxy)-2- oxobenzo[d]ox- azol-3(2H)-
yl)propanoate MH+ 377 1.35 A 46 ##STR00087## 2-amino-2-
(hydroxymethyl) propane-1,3- diol (R)-3-(5- chloro-2-oxo-
6-(1-(pyridin- 2- yl)propoxy)ben- zo[d]oxazol- 3(2H)- yl)propanoate
MH+ 377 1.44 A 46a ##STR00088## (R)-3-(5- chloro-2-oxo-
6-(1-(pyridin- 2- yl)propoxy)ben- zo[d]oxazol- 3(2H)- yl)propanoic
acid MH+ 377 1.44 A 47 ##STR00089## 2-amino-2- (hydroxymethyl)
propane-1,3- diol (R)-3-(5- chloro-6-(1-(4- methylpyridin-
2-yl)ethoxy)-2- oxobenzo[d]ox- azol-3(2H)- yl)propanoate MH+ 377
1.33 A 48 ##STR00090## (R)-3-(5- chloro-2-oxo- 6-(1- (pyridazin-3-
yl)ethoxy)benz- o[d]oxazol- 3(2H)- yl)propanoic acid MH+ 364 1.37 A
49 ##STR00091## (S)-3-(5- chloro-2-oxo- 6-(1- (pyridazin-3-
yl)ethoxy)benz- o[d]oxazol- 3(2H)- yl)propanoic acid MH+ 364 1.36 A
50 ##STR00092## 3-(5-chloro-6- ((6- methylpyridazin- 3-
yl)methoxy)-2- oxobenzo[d]ox- azol-3(2H)- yl)propanoic acid MH+ 364
1.50 A 51 ##STR00093## 3-(5-chloro-6- (1-(6- methylpyridazin-
3-yl)ethoxy)- 2- oxobenzo[d]ox- azol-3(2H)- yl)propanoic acid
(single unknown enantiomer) MH+ 378 1.30 A 52 ##STR00094##
3-(5-chloro-6- (1-(6- methylpyridazin- 3-yl)ethoxy)- 2-
oxobenzo[d]ox- azol-3(2H)- yl)propanoic acid (single unknown
enantiomer) MH+ 378 1.30 A 53 ##STR00095## (R)-3-(5-
chloro-6-(1-(5- methylpyridin- 2-yl)propoxy)- 2- oxobenzo[d]ox-
azol-3(2H)- yl)propanoic acid MH+ 391 1.47 A 54 ##STR00096##
3-(5-chloro-6- (1-(5- methylpyridin- 2-yl)ethoxy)-2- oxobenzo[d]ox-
azol-3(2H)-yl)- 2- methylpropan- oic acid (single unknown
enantiomer) MH+ 391 1.48 A 55 ##STR00097## 3-(5-chloro-6- (1-(5-
methylpyridin- 2-yl)ethoxy)-1- oxobenzo[d]ox- azol-3(2H)-yl)- 2-
methylpropan oic acid (single unknown enantiomer) MH+ 391 1.48 A 56
##STR00098## 3-(5-chloro-6- (1-(5- methylpyridin- 2-yl)ethoxy)-2-
oxobenzo[d]ox- azol-3(2H)-yl)- 2- methylpropan- oic acid (single
unknown enantiomer) MH+ 391 1.48 A 57 ##STR00099## 3-(5-chloro-6-
(1-(5- methylpyridin- 2-yl)ethoxy)-2- oxobenzo[d]ox-
azol-3(2H)-yl)- 2- methylpropan oic acid (single unknown
enantiomer) MH+ 391 1.48 A 58 ##STR00100## 2-amino-2-
(hydroxymethyl) propane-1,3- diol 3-(5- chloro-2-oxo- 6-(1-
(pyrimidin-2- yl)ethoxy)benz- o[d]oxazol- 3(2H)- yl)propanoate
(single unknown enantiomer) MH+ 364 2.55 G 59 ##STR00101##
2-amino-2- (hydroxymethyl) propane-1,3- diol 3-(5- chloro-2-oxo-
6-(1- (pyrimidin-2- yl)ethoxy)benz- o[d]oxazol- 3(2H)-
yl)propanoate (single unknown enaniomer) [M - H].sup.- 362 1.71 F
60 ##STR00102## 2-amino-2- (hydroxymethyl) propane-1,3- diol 3-(5-
chloro-6-(1- (oxazol-2- yl)ethoxy)-2- oxobenzo[d]ox- azol-3(2H)-
yl)propanoate [M - H].sup.- 351 1.83 F 61 ##STR00103## 2-amino-2-
(hydroxymethyl) propane-1,3- diol 3-(5- chloro-6-(1- (oxazol-2-
yl)ethoxy)-2- oxobenzo[d]ox- azol-3(2H)- yl)propanoate [M -
H].sup.- 351 1.83 F
[0535] The potencies and efficacies of the compounds of the
invention for KMO inhibition can be determined by MS Rapidfire
assay performed on the human cloned enzyme as described herein.
Compounds of formula (I) have demonstrated inhibitory activity at
the KMO enzyme, using the MS Rapidfire functional assay described
herein.
[0536] KMO MS Rapidfire Assay Protocol
[0537] Materials and Methods
[0538] Materials
[0539] L-Kynurenine (Kyn), 3-hydroxy-DL-kynurenine (3-HK),
.beta.-Nicotinamide adenine dinucleotide 2'-phosphate reduced
tetrasodium salt hydrate (NADPH),
4-(2-hydroxyethyl)piperazine-1-ethanesulfonic acid (Hepes),
DL-dithiothreitol (DTT), ethylenediaminetetraacetic acid (EDTA),
CHAPS and trifluoroacetic acid (TFA) were purchased from
Sigma-Aldrich Ltd. (Gillingham, Dorset, UK). HPLC-grade
acetonitrile and formic acid were supplied by Fisher Scientific
(Loughborough, UK).
[0540] Cloning and Expression of Human KMO
[0541] Full length human KMO was amplified by PCR from
pcDNA5/FRT/V5-His-TOPO/hKMO (vector supplied by the University of
Edinburgh) and cloned into pGEX6P-1 (GE Healthcare) using BamH1 and
Sal1 restriction sites. DNA encoding the N-terminal
Glutathione-S-transferase (GST) tag, followed by a Pre-Scission
protease cleavage site, and the full length KMO was amplified by
PCR from pGEX6P-1-KMO and cloned into pFastbac1 (Invitrogen) using
Xbal and EcoR1 restriction sites.
[0542] pFastbac1 GST-KMO was transposed into the baculovirus genome
using the BAC-to-BAC technology (Invitrogen) and bacmid DNA was
prepared and transfected into Spodoptera frugiperda (Sf9) cells
using Cellfectin II (Invitrogen). Expression of a protein of the
expected molecular weight (Mr 82,634) was seen by Western blot
analysis using anti-GST-peroxidase conjugate.
[0543] Preparation of Membranes from Sf9 Cells Expressing Human
GST-KMO
[0544] A P1 virus stock was generated from a single clone and used
to infect 3.times.1.5 L cultures of Sf9 cells in 3 L Corning
Fernbach flasks. The Sf9 cells were grown in Hyclone SFX media
(Thermo Scientific) to about 3.times.10.sup.6 cells/ml and were
infected at a nominal multiplicity of infection of 3. Cells were
harvested after 48 hours and disrupted by blending in 50 mM Hepes,
pH 7.4, 1 mM EDTA buffer containing protease inhibitors. A low
speed spin (400 g) was used to remove cell debris, followed by a
high speed spin (75 000 g) to pellet the membranes. The membranes
were purified in a discontinuous sucrose density gradient by
re-suspending in 10% (w/v) sucrose and layering over 40% (w/v)
sucrose, both in the above buffer. This was centrifuged at 150 000
g and the purified membranes were taken from the interface,
collected by centrifugation at 100 000 g, resuspended in buffer and
aliquoted for storage at -80.degree. C. KMO activity was found to
be associated with the membrane fraction only and no KMO activity
was detected in membranes prepared from uninfected Sf9 cells. A
batch of 104 mg of purified Sf9 KMO-membranes (as determined by the
Pierce BCA protein assay using bovine serum albumin as standard)
was prepared and validated in the RF MS assay.
[0545] RapidFire High-Throughput Mass Spectrometry Assay
[0546] 11 point, 3-fold serial dilutions of test compounds were
prepared in DMSO and 100 nL of these solutions were dispensed into
384-well V-base polypropylene plates (Greiner Bio-one, Stonehouse,
UK) using an Echo 555 acoustic dispenser (Labcyte, Sunnyvale,
Calif.). This gave a final assay concentration range between 100
.mu.M and 1.7 nM in 10 .mu.L final assay volume (see below). 100 nL
DMSO was dispensed into columns 6 and 18 for high and low controls,
respectively, with prior inactivation of the enzyme in column 18 by
pre-dispense of 30 .mu.L of 0.5% (v/v) TFA.
[0547] Conditions for the assay of human KMO using isolated
KMO-membranes were 50 mM Hepes, pH 7.5, 2 mM DTT, 1 mM EDTA, 100
.mu.M CHAPS, 200 .mu.M NADPH, 10 .mu.M Kynurenine and 8 .mu.g/ml
KMO-membranes in a total reaction volume of 10 .mu.L.
[0548] Assays were performed by initially dispensing 5 .mu.L of a
2.times. Enzyme solution (16 .mu.g/ml KMO-membranes in 50 mM Hepes,
pH 7.5, 2 mM DTT, 2 mM EDTA, 200 .mu.M CHAPS) into plates
containing 100 nL compounds and incubating for 10 min at ambient
temperature. Reactions were initiated by addition of 5 .mu.L of
2.times. Substrate solution (400 .mu.M NADPH, 20 .mu.M Kynurenine
in 50 mM Hepes, pH 7.5, 2 mM DTT) and incubated for 2 h at room
temperature before quenching the reaction with 30 .mu.L of 0.5%
(v/v) TFA. Plates were centrifuged at 2500 rpm for 10 min before
analysis. All additions were made using a Multidrop Combi dispenser
(Thermo Fisher Scientific).
[0549] Quenched assay plates were transferred to a high-throughput
RapidFire200 integrated autosampler/solid-phase extraction (SPE)
system (Agilent Technologies, Wakefield, Mass.). Samples were
aspirated from each well for 500 ms and 10 .mu.L was loaded
directly onto a RapidFire micro-scale SPE C18 (type C) cartridge,
which was washed for 3 s with HPLC-grade water containing 0.1%
(v/v) formic acid to remove non-organic components. Analytes were
then eluted into the mass spectrometer, in a 3 s elution cycle,
using 80% (v/v) acetonitrile/water containing 0.1% (v/v) formic
acid, and the cartridge was then equilibrated by washing with water
containing 0.1% (v/v) formic acid for 500 ms. This gave a total
cycle time of 7 s, enabling analysis of a 384-well plate in
approximately 45 min.
[0550] Both Kyn and 3-HK were detected using a Sciex API4000 triple
quadrupole mass spectrometer (Applied Biosystems, Concord, Ontario,
Canada), equipped with an electrospray interface and operated in
positive ion mode. Multiple reaction monitoring (MRM) was used to
detect both Kyn and 3-HK using Q1/Q3 transitions at m/z 209.4 to
192.0 and m/z 225.3 to 208.2, respectively. The mass spectrometer
used an ESI voltage of 5500 V and a source temperature of
600.degree. C., with a dwell time of 50 ms for each transition.
[0551] Data Analysis
[0552] Individual MRM transitions were saved as text files and the
extracted ion chromatograms were integrated and processed using the
RapidFire.RTM. peak integration software (version 3.6).
[0553] Using the integrated peak area for 3-HK data was analysed
within ActivityBase (ID Business Solutions Ltd, Surrey, UK). Dose
response curves were fitted to equation (1):
Inhibition ( % ) = ( a - d ) 1 + ( [ I ] IC 50 ) S + d ( 1 )
##EQU00001##
[0554] Where a is the uninhibited response, d is the fully
inhibited response, [I] is the inhibitor concentration, IC.sub.50
is [I] that gives 0.5.times.(a-d) and S is the Hill slope.
[0555] Exemplified compounds of the invention have median
pIC.sub.50 values of >6.1 in the above MS Rapidfire assay.
Example 1 has a median pIC.sub.50=7.9 in the above MS Rapidfire
assay. Example 2 has a median pIC.sub.50=8.4 in the above MS
Rapidfire assay.
* * * * *