U.S. patent application number 17/325068 was filed with the patent office on 2021-12-09 for inhibitors of alpha-amino-beta-carboxymuconic acid semialdehyde decarboxylase.
The applicant listed for this patent is TES Pharma S.r.l.. Invention is credited to Francesca DE FRANCO, Nicola GIACCHE, Paride LISCIO, Roberto PELLICCIARI.
Application Number | 20210379069 17/325068 |
Document ID | / |
Family ID | 1000005829904 |
Filed Date | 2021-12-09 |
United States Patent
Application |
20210379069 |
Kind Code |
A1 |
PELLICCIARI; Roberto ; et
al. |
December 9, 2021 |
INHIBITORS OF ALPHA-AMINO-BETA-CARBOXYMUCONIC ACID SEMIALDEHYDE
DECARBOXYLASE
Abstract
The present disclosure discloses compounds capable of modulating
the activity of .alpha.-amino-.beta.-carboxymuconic acid
semialdehyde decarboxylase (ACMSD), which are useful for the
prevention and/or the treatment of diseases and disorders
associated with defects in NAD.sup.+ biosynthesis, e.g., metabolic
disorders, neurodegenerative diseases, chronic inflammatory
diseases, kidney diseases, and diseases associated with ageing. The
present application also discloses pharmaceutical compositions
comprising said compounds and the use of such compounds as a
medicament.
Inventors: |
PELLICCIARI; Roberto;
(Perugia, IT) ; LISCIO; Paride; (Perugia, IT)
; GIACCHE; Nicola; (Perugia, IT) ; DE FRANCO;
Francesca; (Perugia, IT) |
|
Applicant: |
Name |
City |
State |
Country |
Type |
TES Pharma S.r.l. |
Perugia |
|
IT |
|
|
Family ID: |
1000005829904 |
Appl. No.: |
17/325068 |
Filed: |
May 19, 2021 |
Related U.S. Patent Documents
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Application
Number |
Filing Date |
Patent Number |
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PCT/EP2019/081799 |
Nov 19, 2019 |
|
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17325068 |
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62769959 |
Nov 20, 2018 |
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Current U.S.
Class: |
1/1 |
Current CPC
Class: |
C07D 409/14 20130101;
A61K 31/4436 20130101; A61K 31/5377 20130101; A61K 31/513 20130101;
C07D 409/12 20130101; A61P 1/16 20180101; C07D 409/04 20130101;
C07D 213/70 20130101; C07D 239/56 20130101; A61K 31/4418
20130101 |
International
Class: |
A61K 31/513 20060101
A61K031/513; C07D 409/04 20060101 C07D409/04; C07D 409/14 20060101
C07D409/14; C07D 213/70 20060101 C07D213/70; C07D 409/12 20060101
C07D409/12; C07D 239/56 20060101 C07D239/56; A61P 1/16 20060101
A61P001/16; A61K 31/5377 20060101 A61K031/5377; A61K 31/4418
20060101 A61K031/4418; A61K 31/4436 20060101 A61K031/4436 |
Claims
1. A compound represented by Formula (II): ##STR00400## or a
pharmaceutically acceptable salt or tautomer thereof, wherein: X is
H, S, SR.sup.2, NR.sup.2, NR.sup.2R.sup.2', O, OH, OR.sup.h, F, Br,
or C.sub.1; W is N or C; (i) when W is N, then: L is
--(C(R.sup.5).sub.2).sub.mCH.dbd.CH(C(R.sup.5).sub.2).sub.p--,
##STR00401##
--(C(R.sup.5).sub.2).sub.mY.sup.1(C(R.sup.5).sub.2).sub.p--,
--(C(R.sup.5).sub.2).sub.mY.sup.1(C(R.sup.5).sub.2).sub.p-cyclopropyl-,
--(C(R.sup.5).sub.2).sub.mY.sup.1CH.dbd.CH--,
--(C(R.sup.5).sub.2).sub.mNR.sup.3C.dbd.(O)(C(R.sup.5).sub.2).sub.p--,
--(C(R.sup.5).sub.2).sub.mphenyl(C(R.sup.5).sub.2).sub.p--,
--(C(R.sup.5).sub.2).sub.mpyridinyl(C(R.sup.5).sub.2).sub.p--, or
--(C(R.sup.5).sub.2).sub.mthiophenyl(C(R.sup.5).sub.2).sub.p--;
(ii) when W is C, then: L is
--(C(R.sup.5).sub.2).sub.mCH.dbd.CH(C(R.sup.5).sub.2).sub.p--,
--(C(R.sup.5).sub.2).sub.o--,
--(C(R.sup.5).sub.2).sub.mY.sup.1(C(R.sup.5).sub.2).sub.p--,
##STR00402## --(C(R.sup.5).sub.2).sub.m Y.sup.1CH.dbd.CH--,
--(C(R.sup.5).sub.2).sub.mC.dbd.(O)(CH.sub.2).sub.p--,
--(C(R.sup.5).sub.2).sub.mC.dbd.(O)O(C(R.sup.5).sub.2).sub.p--,
--(C(R.sup.5).sub.2).sub.mC.dbd.(O)NR.sup.3(C(R.sup.5).sub.2).sub.p--,
--(C(R.sup.5).sub.2).sub.mNR.sup.3C.dbd.(O)(C(R.sup.5).sub.2).sub.p--,
--(C(R.sup.5).sub.2).sub.mphenyl(C(R.sup.5).sub.2).sub.p--,
--(C(R.sup.5).sub.2).sub.mpyridinyl(C(R.sup.5).sub.2).sub.p--, or
--(C(R.sup.5).sub.2).sub.mthiophenyl(C(R.sup.5).sub.2).sub.p--;
Y.sup.1 is O, NR.sup.4, or S(O).sub.q; each Y.sup.2 is
independently O, NH or S; R.sup.1 is absent, C.sub.6-C.sub.10
arylene, heteroarylene, or C.sub.3-C.sub.8cycloalkylene, wherein
the heteroarylene comprises one or two 5- to 7-membered rings and
1-4 heteroatoms selected from N, O and S, and wherein the
C.sub.6-C.sub.10 arylene, heteroarylene, and
C.sub.3-C.sub.8cycloalkylene are optionally substituted with one to
two R.sup.e; R.sup.2 is H or C.sub.1-C.sub.4 alkyl; R.sup.2' is H,
C.sub.1-C.sub.4 alkyl, or C.sub.3-C.sub.7 cycloalkyl; or R.sup.2
and R.sup.2' together with the nitrogen atom to which they are
attached form a 3- to 7-membered heterocycloalkyl ring comprising
1-3 additional heteroatoms selected from N, O and S; R.sup.3 is H
or C.sub.1-C.sub.4 alkyl; R.sup.4 is H or C.sub.1-C.sub.4 alkyl;
each R.sup.5 is independently at each occurrence H or
C.sub.1-C.sub.4 alkyl; each R.sup.6 is independently at each
occurrence H or C.sub.1-C.sub.4 alkyl; R.sup.7 is H, A, B, or C; A
is --(C(R.sup.6).sub.2).sub.rCO.sub.2R.sup.x,
--Y.sup.2(C(R.sup.6).sub.2).sub.rCO.sub.2R.sup.x,
--(C(R.sup.6).sub.2).sub.rtetrazole,
--(C(R.sup.6).sub.2).sub.roxadiazolone,
--(C(R.sup.6).sub.2).sub.rtetrazolone,
--(C(R.sup.6).sub.2).sub.rthiadiazolol, --(C(R.sup.6).sub.2).sub.r
isoxazol-3-ol, --(C(R.sup.6).sub.2).sub.rP(O)(OH)OR.sup.x,
--(C(R.sup.6).sub.2).sub.rS(O).sub.2OH,
--(C(R.sup.6).sub.2).sub.rC(O)NHCN, or
--(C(R.sup.6).sub.2).sub.rC(O)NHS(O).sub.2alkyl, wherein
--(C(R.sup.6).sub.2).sub.rtetrazole,
--(C(R.sup.6).sub.2).sub.roxadiazolone,
--(C(R.sup.6).sub.2).sub.rtetrazolone,
--(C(R.sup.6).sub.2).sub.rthiadiazolol, --(C(R.sup.6).sub.2).sub.r
isoxazol-3-ol are optionally substituted with C.sub.1-C.sub.6
alkyl, B is --(C(R.sup.6).sub.2).sub.rS(O).sub.2OC.sub.1-C.sub.4
alkyl, --O(C(R.sup.6).sub.2).sub.rS(O).sub.2OC.sub.1-C.sub.4 alkyl,
--Y.sup.2(C(R.sup.6).sub.2).sub.rC(O)NR.sup.gR.sup.g',
--Y.sup.2(C(R.sup.6).sub.2).sub.rS(O).sub.2NR.sup.gR.sup.g',
--(C(R.sup.6).sub.2).sub.rC(O)NR.sup.gR.sup.g',
--(C(R.sup.6).sub.2).sub.rS(O).sub.2NR.sup.gR.sup.g',
--(C(R.sup.6).sub.2).sub.rC(O)NHS(O).sub.2NR.sup.gR.sup.g',
--(C(R.sup.6).sub.2).sub.rCO.sub.2R.sup.r,
--(C(R.sup.6).sub.2).sub.rNH.sub.2CO.sub.2R.sup.x,
--(C(R.sup.6).sub.2).sub.rP(O)(OR.sup.x).sub.2,
--O(C(R.sup.6).sub.2).sub.rP(O)(OR.sup.x).sub.2,
--(C(R.sup.6).sub.2).sub.rS(O).sub.2OH,
--O(C(R.sup.6).sub.2).sub.rS(O).sub.2OH,
--(C(R.sup.6).sub.2).sub.rP(O).sub.2OR.sup.x, or
--O(C(R.sup.6).sub.2).sub.rP(O).sub.2OR.sup.x, C is
--(CH.sub.2).sub.rCN, --(CH.sub.2).sub.sOH, halogen,
--(C(R)).sub.2).sub.rC.sub.6-C.sub.10 aryl,
--(C(R.sup.6).sub.2).sub.rS--C.sub.6-C.sub.10 aryl,
--(C(R.sup.6).sub.2).sub.rheteroaryl,
--O(C(R.sup.6).sub.2).sub.rheteroaryl,
--O(C(R.sup.6).sub.2).sub.rheterocycloalkyl,
--O(C(R.sup.6).sub.2).sub.rOH, --OR.sup.y,
--(C(R.sup.6).sub.2).sub.rC(O)NHCN, --CH.dbd.CHCO.sub.2R.sup.x, or
--(C(R.sup.6).sub.2).sub.rC(O)NHS(O).sub.2C.sub.1-C.sub.4 alkyl,
wherein the aryl and heteroaryl are substituted with one to three
substituents each independently selected from C.sub.1-C.sub.6
alkyl, C.sub.1-C.sub.6 haloalkyl, halogen, and OH, and wherein the
heterocycloalkyl is substituted with one to two .dbd.O or .dbd.S;
R.sup.c is H, C.sub.1-C.sub.6 alkyl, C.sub.1-C.sub.6 haloalkyl,
halogen, --CN, --OR.sup.x, or --CO.sub.2R.sup.x; R.sup.d is methyl,
CF.sub.3, CR.sup.fF.sub.2, --(C(R.sup.6).sub.2).sub.tC6-C.sub.10
aryl, --(C(R.sup.6).sub.2).sub.t-5- or 6-membered heteroaryl,
--(C(R.sup.6).sub.2).sub.t-5- or 6-membered cycloalkyl, optionally
substituted C.sub.6-C.sub.10 aryl, optionally substituted 5- or
6-membered heteroaryl, or optionally substituted 5- or 6-membered
cycloalkyl; each R.sup.e is independently at each occurrence
C.sub.1-C.sub.6 alkyl, C.sub.2-C.sub.6 alkenyl, C.sub.2-C.sub.6
alkynyl, halogen, C.sub.1-C.sub.6 haloalkyl, --NHR.sup.z, --OH, or
--CN; R.sup.f is absent, H, or methyl; R.sup.g is H,
C.sub.1-C.sub.6 alkyl, OH, --S(O).sub.2(C.sub.1-C.sub.6 alkyl), or
S(O).sub.2N(C.sub.1-C.sub.6 alkyl).sub.2; R.sup.g' is H,
C.sub.1-C.sub.6 alkyl, C.sub.3-C.sub.7 cycloalkyl, 4- to 7-membered
heterocycloalkyl ring comprising 1-3 heteroatoms selected from N, O
and S, C.sub.6-C.sub.10 aryl, or 5- to 7-membered heteroaryl
comprising 1-3 heteroatoms selected from N, O and S, wherein the
alkyl is optionally substituted with one or more substituents
independently selected from halogen and --OH, and wherein the
cycloalkyl, heterocycloalkyl, aryl and heteroaryl are optionally
substituted with one or more substituents independently selected
from C.sub.1-C.sub.6 alkyl, halogen, and --OH; R.sup.h is H,
C.sub.1-C.sub.4 alkyl, or 3- to 7-membered heterocycloalkyl ring
comprising 1-3 heteroatoms selected from N, O and S, wherein the
alkyl is optionally substituted with one or more substituents each
independently selected from NH2, C.sub.1-C.sub.4 alkylamino,
C.sub.1-C.sub.4 dialkylamino, and C(O)NH2; and wherein the
heterocycloalkyl is optionally substituted with one or more
substituents each independently selected from C.sub.1-C.sub.6 alkyl
and C.sub.1-C.sub.6 haloalkyl; R.sup.i is (i)
--(CH.sub.2).sub.sOC(O)C.sub.1-C.sub.6 alkyl, wherein the alkyl is
substituted with one or more NH2; (ii)
(CH.sub.2CH.sub.2O).sub.nCH2CH.sub.2OH; or (iii) C.sub.1-C.sub.6
alkyl substituted with one or more substituents each independently
selected from OH and 4- to 7-membered heterocycloalkyl comprising 1
to 3 heteroatoms selected from O, N, or S; R.sup.j is absent, H,
C.sub.1-C.sub.6 alkyl, or --CN; each R.sup.x is independently at
each occurrence H, C.sub.1-C.sub.6 alkyl, or C.sub.6-C.sub.10 aryl;
each R.sup.y and R.sup.z is independently H, C.sub.1-C.sub.6 alkyl,
or C.sub.1-C.sub.6 haloalkyl; each m, p, q, r, and t is
independently 0, 1 or 2; n is 0, 1, 2, or 3; s is 1 or 2; is 0, 1,
2, 3, or 4; and represents a single bond or a double bond; and
provided that when X is O; R.sup.f is H; W is C; R.sup.j is --CN; L
is --SCH.sub.2--; R.sup.1 is phenylene or pyridine; then R.sup.7 is
not --COOH; when X is O; R.sup.f is H; W is C; R.sup.i is --CN; L
is --SCH.sub.2--; R.sup.1 is phenylene or pyridine; and R.sup.7 is
tetrazole; then R.sup.c is not H; when X is O; R.sup.f is H; W is
C; R.sup.i is --CN; L is --S--C(R.sup.5).sub.2 or
--SCH.sub.2CH.sub.2--; R.sup.1 is absent; then R.sup.7 is not COOH
or tetrazole; when X is O, R.sup.f is H; W is N; R.sup.i is absent;
R.sup.d is methyl, optionally substituted 5- to 10-membered aryl,
optionally substituted 5- or 6-membered heteroaryl, or optionally
substituted 5- or 6-membered cycloalkyl; L is --SCH.sub.2-- or
--OCH.sub.2--; and R.sup.1 is phenylene; then R.sup.7 is not
--COOH, --CH.sub.2COOH, ##STR00403## and when X is O, R.sup.f is H,
W is N, R.sup.i is absent, L is --NHCH.sub.2--, --CH.sub.2NH--, or
--NH--C(O)--, and R.sup.1 is phenylene, then R.sup.d is not
phenyl.
2. A compound represented by Formula (I): ##STR00404## or a
pharmaceutically acceptable salt or tautomer thereof, wherein: X is
H, S, SR.sup.2, NR.sup.2, NR.sup.2R.sup.2', O, OH, OR.sup.h, F, Br,
or C.sub.1; W is N or C; (i) when W is N, then: L is
--(C(R.sup.5).sub.2).sub.mCH.dbd.CH(C(R.sup.5).sub.2).sub.p--,
##STR00405##
--(C(R.sup.5).sub.2).sub.mY.sup.1(C(R.sup.5).sub.2).sub.p--,
--(C(R.sup.5).sub.2).sub.mY.sup.1(C(R.sup.5).sub.2).sub.p-cyclopropyl-,
--(C(R.sup.5).sub.2).sub.mY.sup.1CH.dbd.CH--,
--(C(R.sup.5).sub.2).sub.mNR.sup.3C.dbd.(O)(C(R.sup.5).sub.2).sub.p--,
--(C(R.sup.5).sub.2).sub.mphenyl(C(R.sup.5).sub.2).sub.p--,
--(C(R.sup.5).sub.2).sub.mpyridinyl(C(R.sup.5).sub.2).sub.p--, or
--(C(R.sup.5).sub.2).sub.mthiophenyl(C(R.sup.5).sub.2).sub.p--;
(ii) when W is C, then: L is
--(C(R.sup.5).sub.2).sub.mCH.dbd.CH(C(R.sup.5).sub.2).sub.p--,
--(C(R.sup.5).sub.2).sub.o--,
--(C(R.sup.5).sub.2).sub.mY.sup.1(C(R.sup.5).sub.2).sub.p--,
##STR00406## --(C(R.sup.5).sub.2).sub.mY.sup.1CH.dbd.CH--,
--(C(R.sup.5).sub.2).sub.mC.dbd.(O)(CH.sub.2).sub.p--,
--(C(R.sup.5).sub.2).sub.mC.dbd.(O)O(C(R.sup.5).sub.2).sub.p--,
--(C(R.sup.5).sub.2).sub.mC.dbd.(O)NR.sup.3(C(R.sup.5).sub.2).sub.p--,
--(C(R.sup.5).sub.2).sub.mNR.sup.3C.dbd.(O)(C(R.sup.5).sub.2).sub.p--,
--(C(R.sup.5).sub.2).sub.mphenyl(C(R.sup.5).sub.2).sub.p--,
--(C(R.sup.5).sub.2).sub.mpyridinyl(C(R.sup.5).sub.2).sub.p--, or
--(C(R.sup.5).sub.2).sub.mthiophenyl(C(R.sup.5).sub.2).sub.p--;
Y.sup.1 is O, NR.sup.4, or S(O).sub.q; each Y.sup.2 is
independently O, NH or S; R.sup.1 is absent or C.sub.6-C.sub.10
arylene or heteroarylene, wherein the heteroarylene comprises one
or two 5- to 7-membered rings and 1-4 heteroatoms selected from N,
O and S, and wherein the C.sub.6-C.sub.10 arylene or heteroarylene
are optionally substituted with one to two R.sup.e; R.sup.2 is H or
C.sub.1-C.sub.4 alkyl; R.sup.2' is H, C.sub.1-C.sub.4 alkyl, or
C.sub.3-C.sub.7 cycloalkyl; or R.sup.2 and R.sup.2' together with
the nitrogen atom to which they are attached form a 3- to
7-membered heterocycloalkyl ring comprising 1-3 additional
heteroatoms selected from N, O and S; R.sup.3 is H or
C.sub.1-C.sub.4 alkyl; R.sup.4 is H or C.sub.1-C.sub.4 alkyl; each
R.sup.s is independently at each occurrence H or C.sub.1-C.sub.4
alkyl; each R.sup.6 is independently at each occurrence H or
C.sub.1-C.sub.4 alkyl; R.sup.7 is H, A, B, or C; A is
--(C(R.sup.6).sub.2).sub.rCO.sub.2R.sup.x,
--Y.sup.2(C(R.sup.6).sub.2).sub.rCO.sub.2R.sup.x,
--(CH.sub.2).sub.rtetrazole, --(CH.sub.2).sub.roxadiazolone,
--(CH.sub.2).sub.rtetrazolone, --(CH.sub.2).sub.rthiadiazolol,
--(CH.sub.2).sub.r isoxazol-3-ol,
--(CH.sub.2).sub.rP(O)(OH)OR.sup.x, --(CH.sub.2).sub.rS(O).sub.2OH,
--(CH.sub.2).sub.rC(O)NHCN, or
--(CH.sub.2).sub.rC(O)NHS(O).sub.2alkyl, wherein
--(CH.sub.2).sub.rtetrazole, --(CH.sub.2).sub.roxadiazolone,
--(CH.sub.2).sub.rtetrazolone, --(CH.sub.2).sub.rthiadiazolol,
--(CH.sub.2).sub.r isoxazol-3-ol are optionally substituted with
C.sub.1-C.sub.6 alkyl, B is
--(C(R.sup.6).sub.2).sub.rS(O).sub.2OC.sub.1-C.sub.4 alkyl,
--O(C(R.sup.6).sub.2).sub.rS(O).sub.2OC.sub.1-C.sub.4 alkyl,
--Y.sup.2(C(R.sup.6).sub.2).sub.rC(O)NR.sup.gR.sup.g',
--Y.sup.2(C(R.sup.6).sub.2).sub.rS(O).sub.2NR.sup.gR.sup.g',
--(CH.sub.2).sub.rC(O)NR.sup.gR.sup.g',
--(CH.sub.2).sub.rS(O).sub.2NR.sup.gR.sup.g',
--(CH.sub.2).sub.rC(O)NHS(O).sub.2NR.sup.gR.sup.g',
--(C(R.sup.6).sub.2).sub.rCO.sub.2R.sup.i,
--(C(R.sup.6).sub.2).sub.rNH.sub.2CO.sub.2R.sup.x,
--(C(R.sup.6).sub.2).sub.rP(O)(OR.sup.x).sub.2,
--O(C(R.sup.6).sub.2).sub.rP(O)(OR.sup.x).sub.2,
--(C(R.sup.6).sub.2).sub.rS(O).sub.2OH,
--O(C(R.sup.6).sub.2).sub.rS(O).sub.2OH,
--(C(R.sup.6).sub.2).sub.rP(O).sub.2OR.sup.x, or
--O(C(R.sup.6).sub.2).sub.rP(O).sub.2OR.sup.x, C is
--(CH.sub.2).sub.rCN, --(CH.sub.2).sub.sOH, halogen,
--(C(R.sup.6).sub.2).sub.rC.sub.6-C.sub.10 aryl,
--(C(R.sup.6).sub.2).sub.rS--C.sub.6-C.sub.10 aryl,
--(C(R.sup.6).sub.2).sub.rheteroaryl,
--O(C(R.sup.6).sub.2).sub.rheteroaryl,
--O(C(R.sup.6).sub.2).sub.rheterocycloalkyl,
--O(C(R.sup.6).sub.2).sub.rOH, --OR.sup.y,
--(C(R.sup.6).sub.2).sub.rC(O)NHCN, --CH.dbd.CHCO.sub.2R.sup.x, or
--(C(R.sup.6).sub.2).sub.rC(O)NHS(O).sub.2C.sub.1-C.sub.4 alkyl,
wherein the aryl and heteroaryl are substituted with one to three
substituents each independently selected from C.sub.1-C.sub.6
alkyl, C.sub.1-C.sub.6 haloalkyl, halogen, and OH, and wherein the
heterocycloalkyl is substituted with one to two .dbd.O or .dbd.S;
R.sup.c is H, C.sub.1-C.sub.6 alkyl, C.sub.1-C.sub.6 haloalkyl,
halogen, --CN, --OR.sup.x, or --CO.sub.2R.sup.x; R.sup.d is methyl,
CF.sub.3, CR.sup.fF.sub.2,
--(C(R.sup.6).sub.2).sub.tC.sub.6-C.sub.10 aryl,
--(C(R.sup.6).sub.2).sub.t-5- or 6-membered heteroaryl,
--(C(R.sup.6).sub.2).sub.t-5- or 6-membered cycloalkyl, optionally
substituted C.sub.6-C.sub.10 aryl, optionally substituted 5- or
6-membered heteroaryl, or optionally substituted 5- or 6-membered
cycloalkyl; each R.sup.e is independently at each occurrence
C.sub.1-C.sub.6 alkyl, C.sub.2-C.sub.6 alkenyl, C.sub.2-C.sub.6
alkynyl, halogen, C.sub.1-C.sub.6 haloalkyl, --NHR.sup.z, --OH, or
--CN; R.sup.f is absent, H, or methyl; R.sup.g is H,
C.sub.1-C.sub.6 alkyl, OH, --S(O).sub.2(C.sub.1-C.sub.6 alkyl), or
S(O).sub.2N(C.sub.1-C.sub.6 alkyl).sub.2; R.sup.g' is H,
C.sub.1-C.sub.6 alkyl, C.sub.3-C.sub.7 cycloalkyl, 4- to 7-membered
heterocycloalkyl ring comprising 1-3 heteroatoms selected from N, O
and S, C.sub.6-C.sub.10 aryl, or 5- to 7-membered heteroaryl
comprising 1-3 heteroatoms selected from N, O and S, wherein the
alkyl is optionally substituted with one or more substituents
independently selected from halogen and --OH, and wherein the
cycloalkyl, heterocycloalkyl, aryl and heteroaryl are optionally
substituted with one or more substituents independently selected
from C.sub.1-C.sub.6 alkyl, halogen, and --OH; R.sup.h is H,
C.sub.1-C.sub.4 alkyl, or 3- to 7-membered heterocycloalkyl ring
comprising 1-3 heteroatoms selected from N, O and S, wherein the
alkyl is optionally substituted with one or more substituents each
independently selected from NH.sub.2, C.sub.1-C.sub.4 alkylamino,
C.sub.1-C.sub.4 dialkylamino, and C(O)NH.sub.2; and wherein the
heterocycloalkyl is optionally substituted with one or more
substituents each independently selected from C.sub.1-C.sub.6 alkyl
and C.sub.1-C.sub.6 haloalkyl; R.sup.i is (i)
--(CH.sub.2).sub.sOC(O)C.sub.1-C.sub.6 alkyl, wherein the alkyl is
substituted with one or more NH.sub.2; (ii)
(CH.sub.2CH.sub.2O).sub.nCH2CH.sub.2OH; or (iii) C.sub.1-C.sub.6
alkyl substituted with one or more substituents each independently
selected from OH and 4- to 7-membered heterocycloalkyl comprising 1
to 3 heteroatoms selected from O, N, or S; R.sup.i is absent, H,
C.sub.1-C.sub.6 alkyl, or --CN; each R.sup.x is independently at
each occurrence H, C.sub.1-C.sub.6 alkyl, or C.sub.6-C.sub.10 aryl;
each R.sup.y and R.sup.z is independently H, C.sub.1-C.sub.6 alkyl,
or C.sub.1-C.sub.6 haloalkyl; each m, p, q, r, and t is
independently 0, 1 or 2; n is 0, 1, 2, or 3; s is 1 or 2; is 0, 1,
2, 3, or 4; and represents a single bond or a double bond; and
provided that when X is O; R.sup.f is H; W is C; R.sup.i is --CN; L
is --SCH.sub.2--; R.sup.1 is phenylene or pyridine; then R.sup.7 is
not --COOH; when X is O; R.sup.f is H; W is C; R.sup.i is --CN; L
is --SCH.sub.2--; R.sup.1 is phenylene or pyridine; and R.sup.7 is
tetrazole; then R.sup.c is not H; when X is O; R.sup.f is H; W is
C; R.sup.i is --CN; L is --S--C(R.sup.5).sub.2 or
--SCH.sub.2CH.sub.2--; R.sup.1 is absent; then R.sup.7 is not COOH
or tetrazole; when X is O, R.sup.f is H; W is N; R.sup.i is absent;
R.sup.d is methyl, optionally substituted 5- to 10-membered aryl,
optionally substituted 5- or 6-membered heteroaryl, or optionally
substituted 5- or 6-membered cycloalkyl; L is --SCH.sub.2-- or
--OCH.sub.2--; and R.sup.1 is phenylene; then R.sup.7 is not
--COOH, --CH.sub.2COOH, ##STR00407## and when X is O, R.sup.f is H,
W is N, R.sup.j is absent, L is --NHCH.sub.2--, --CH.sub.2NH--, or
--NH--C(O)--, and R.sup.1 is phenylene, then R.sup.d is not
phenyl.
3. The compound of claim 1, wherein X is O, OH, OR.sup.h, F, Br, or
Cl.
4. The compound of claim 1, wherein X is H, S, SR.sup.2, NR.sup.2,
or NR.sup.2R.sup.2'.
5. The compound of claim 1, wherein R.sup.f is absent.
6. The compound of claim 1, wherein R.sup.f is H or methyl.
7. The compound of claim 1, wherein W is N.
8. The compound of claim 7, wherein R.sup.j is absent.
9. The compound of claim 1, wherein W is C.
10. The compound of claim 9, wherein R.sup.j is H, C.sub.1-C.sub.6
alkyl, or --CN.
11. The compound of claim 9, wherein R.sup.j is --CN.
12. The compound of claim 1, wherein R.sup.c is C.sub.1-C.sub.6
alkyl, --CN, or halogen.
13. The compound of claim 1, wherein R.sup.c is --CN or
halogen.
14. The compound of claim 1, wherein R.sup.c is --CN.
15. The compound of claim 1, wherein R.sup.d is methyl.
16. The compound of claim 1, wherein R.sup.d is optionally
substituted 5- to 10-membered aryl.
17. The compound of claim 1, wherein R.sup.d is optionally
substituted 5- or 6-membered heteroaryl.
18. The compound of claim 1, wherein R.sup.d is optionally
substituted 5- or 6-membered cycloalkyl.
19. The compound of claim 1, wherein R.sup.d is methyl, cyclohexyl,
pyridinyl, thiazolyl, phenyl, or thienyl.
20. The compound of claim 1, wherein R.sup.d is methyl, cyclohexyl,
pyridinyl, thiazolyl, thienyl, or optionally substituted
phenyl.
21. The compound of claim 1, wherein R.sup.d is methyl.
22. The compound of claim 1, wherein R.sup.d is --CF.sub.3.
23. The compound of claim 1, wherein R.sup.d is
CR.sup.fF.sub.2.
24. The compound of claim 1, wherein R.sup.d is
--(C(R.sup.6).sub.2).sub.rC.sub.6-C.sub.10 aryl,
--(C(R.sup.6).sub.2).sub.r-5- or 6-membered heteroaryl,
--(C(R.sup.6).sub.2).sub.r-5- or 6-membered cycloalkyl.
25. The compound of claim 1, wherein R.sup.d is
--(C(R.sup.6).sub.2).sub.rC.sub.6-C.sub.10 aryl.
26. The compound of claim 1, wherein L is
--(C(R.sup.5).sub.2).sub.mY.sup.1(C(R.sup.5).sub.2).sub.p--.
27. The compound of claim 26, wherein Y.sup.i is S.
28. The compound of claim 1, wherein L is
--(C(R.sup.5).sub.2).sub.mNR.sup.3C.dbd.(O)(C(R.sup.5).sub.2).sub.p--
or
--(C(R.sup.5).sub.2).sub.mY.sup.1(C(R.sup.5).sub.2).sub.p-cyclopropyl-.
29. The compound of claim 1, wherein R.sup.1 is C.sub.6-C.sub.10
arylene.
30. The compound of claim 1, wherein R.sup.1 is heteroarylene.
31. The compound of claim 1, wherein R.sup.1 is absent.
32. The compound of claim 1, wherein R.sup.7 is A.
33. The compound of claim 32, wherein A is
--(C(R.sup.6).sub.2).sub.rCO.sub.2R.sup.x or
--(CH.sub.2).sub.rtetrazole, wherein the
--(CH.sub.2).sub.rtetrazole is optionally substituted with
C.sub.1-C.sub.6 alkyl.
34. The compound of claim 1, wherein R.sup.7 is B.
35. The compound of claim 32, wherein B is
--(CH.sub.2).sub.rC(O)NR.sup.gR.sup.g', or
--(CH.sub.2).sub.rS(O).sub.2NR.sup.gR.sup.g',
36. The compound of claim 1, wherein R.sup.7 is C.
37. The compound of claim 32, wherein C is --(CH.sub.2).sub.rCN,
--(CH.sub.2).sub.sOH, or --(C(R.sup.6).sub.2).sub.rC.sub.6-C.sub.10
aryl, wherein the aryl is substituted with one to three
substituents each independently selected from C.sub.1-C.sub.6
alkyl, C.sub.1-C.sub.6 haloalkyl, halogen, and OH.
38. The compound of claim 1, or a pharmaceutically acceptable salt
or tautomer thereof, selected from the group consisting of:
TABLE-US-00007 Cpd No. Structure I-1 ##STR00408## I-2 ##STR00409##
I-3 ##STR00410## I-4 ##STR00411## I-5 ##STR00412## I-6 ##STR00413##
I-7 ##STR00414## I-8 ##STR00415## I-9 ##STR00416## I-10
##STR00417## I-11 ##STR00418## I-12 ##STR00419## I-13 ##STR00420##
I-14 ##STR00421## I-15 ##STR00422## I-16 ##STR00423## I-17
##STR00424## I-18 ##STR00425## I-19 ##STR00426## I-20 ##STR00427##
I-21 ##STR00428## I-22 ##STR00429## I-23 ##STR00430## I-24
##STR00431## I-25 ##STR00432## I-26 ##STR00433## I-27 ##STR00434##
I-28 ##STR00435## I-29 ##STR00436## I-30 ##STR00437## I-31
##STR00438## I-32 ##STR00439## I-33 ##STR00440## I-34 ##STR00441##
I-35 ##STR00442## I-36 ##STR00443##
39. The compound of claim 1, or a pharmaceutically acceptable salt
or tautomer thereof, selected from the group consisting of:
##STR00444## ##STR00445## ##STR00446##
40. The compound of claim 1, or a pharmaceutically acceptable salt
or tautomer thereof, selected from the group consisting of:
##STR00447## ##STR00448## ##STR00449## ##STR00450## ##STR00451##
##STR00452## ##STR00453## ##STR00454## ##STR00455## ##STR00456##
##STR00457##
41. A pharmaceutical composition comprising a compound of claim 1,
or a pharmaceutically acceptable salt thereof, and at least one of
a pharmaceutically acceptable carrier, diluent, or excipient.
42. The pharmaceutical composition according to claim 41, which
comprises one or more further therapeutic agents.
43. A method of treating, preventing, or reducing the risk of a
disease or disorder inhibited by
.alpha.-amino-.beta.-carboxymuconate-.epsilon.-semialdehyde
decarboxylase (ACMSD) comprising administering to the subject
suffering from or susceptible to developing the disease or disorder
a therapeutically effective amount of one or more compounds of
any-ene claim 1, or a pharmaceutically acceptable salt thereof.
44. A method of treating, preventing, or reducing the risk of a
disease or disorder associated with reduced nicotinamide adenine
dinucleotide (NAD.sup.+) levels comprising administering to the
subject suffering from or susceptible to developing a disease or
disorder associated with reduced NAD.sup.+ levels a therapeutically
effective amount of one or more compounds of claim 1, or a
pharmaceutically acceptable salt thereof.
45. The method of claim 44, wherein the disease is chronic liver
disease selected from primary biliary cirrhosis (PBC),
cerebrotendinous xanthomatosis (CTX), primary sclerosing
cholangitis (PSC), drug induced cholestasis, intrahepatic
cholestasis of pregnancy, parenteral nutrition associated
cholestasis (PNAC), bacterial overgrowth or sepsis associated
cholestasis, autoimmune hepatitis, chronic viral hepatitis,
alcoholic liver disease, nonalcoholic fatty liver disease (NAFLD),
nonalcoholic steatohepatitis (NASH), liver transplant associated
graft versus host disease, living donor transplant liver
regeneration, congenital hepatic fibrosis, choledocholithiasis,
granulomatous liver disease, intra- or extrahepatic malignancy,
Sjogren's syndrome, Sarcoidosis, Wilson's disease, Gaucher's
disease, hemochromatosis, and alpha 1-antitrypsin deficiency.
46. A method of treating a disorder associated with mitochondrial
dysfunction comprising administering to the subject suffering from
or susceptible to developing a metabolic disorder a therapeutically
effective amount of one or more compounds of claim 1, or a
pharmaceutically acceptable salt thereof, that increases
intracellular nicotinamide adenine dinucleotide (NAD.sup.+).
47. The method of claim 46, wherein said disorder associated with
mitochondrial dysfunction is an inherited mitochondrial disease, a
common metabolic disorder, a neurodegenerative disease, an aging
related disorder, a kidney disorder, or a chronic inflammatory
disease.
48. The method of claim 47, wherein the common metabolic disorder
is obesity or type II diabetes.
49. A method of promoting oxidative metabolism comprising
administering to the subject suffering from or susceptible to
developing a metabolic disorder a therapeutically effective amount
of one or more compounds of claim 1, or a pharmaceutically
acceptable salt thereof, that increases intracellular nicotinamide
adenine dinucleotide (NAD.sup.+).
50-80. (canceled)
Description
CROSS REFERENCE TO RELATED APPLICATIONS
[0001] This application claims the benefit of U.S. Provisional
Application No. 62/769,959, filed Nov. 20, 2018, the content of
which is incorporated herein by reference in its entirety.
FIELD OF THE DISCLOSURE
[0002] The present disclosure relates to compounds capable of
modulating the activity of .alpha.-amino-.beta.-carboxymuconic acid
semialdehyde decarboxylase (ACMSD). The compounds of the disclosure
may be used in methods for the prevention and/or the treatment of
diseases and disorders associated with defects in NAD.sup.+
biosynthesis, e.g., metabolic disorders, neurodegenerative
diseases, chronic inflammatory diseases, kidney diseases, and
diseases associated with ageing.
BACKGROUND OF THE DISCLOSURE
[0003] ACMSD is a critical enzyme for tryptophan metabolism, and
regulates NAD.sup.+ biosynthesis from tryptophan. ACMSD is a
zinc-dependent amidohydrolase that participates in picolinic acid
(PA), quinolinic acid (QA) and NAD.sup.+ homeostasis. ACMSD stands
at a branch point of the NAD.sup.+ biosynthetic pathway from
tryptophan and determines the final fate of the amino acid, i.e.,
transformation into PA, complete oxidation through the citric acid
cycle, or conversion into NAD.sup.+ through QA synthesis.
[0004] ACMSD has been purified from liver, kidney, and brain human
tissues. There are two isoforms ACMSD1 and ACMSD2 derived from a
differential splicing of ACMSD gene transcription but only ACMSD1
is endowed with enzymatic activity. ACMSD1 directs ACMS
(.alpha.-amino-.omega.-carboxymuconic acid semialdehyde) to the
acetyl-CoA pathway, and when ACMSD1 is inhibited, ACMS is
non-enzymatically converted to quinolinic acid (QA) leading to the
formation of NAD.sup.+ and an increase in the intracellular level
of NAD.sup.+.
[0005] Increased levels of NAD.sup.+ have been shown to protect
against neuronal degeneration, improve muscle function and
oxidative metabolism in mice, and enhance lifespan in worms. Whilst
reduced levels of NAD.sup.+ have been associated with a range of
pathophysiological states including type 2 diabetes (T2D),
hyperlipidemia (elevated cholesterol and TAGs), mitochondrial
diseases, neutropenia, cancers, and kidney disorders.
[0006] The inhibition of ACMSD thus represents a novel approach to
increase NAD.sup.+ levels and modify disease pathophysiologies
associated with defects in NAD.sup.+ biosynthesis.
SUMMARY OF THE DISCLOSURE
[0007] It is an object of embodiments of the disclosure to provide
novel series of compounds capable of modulating the activity of
.alpha.-amino-.beta.-carboxymuconic acid semialdehyde decarboxylase
(ACMSD), which compounds are useful for the prevention and/or the
treatment of diseases and disorders associated with defects in
NAD.sup.+ biosynthesis, e.g., metabolic disorders,
neurodegenerative diseases, chronic inflammatory diseases, kidney
diseases, and diseases associated with ageing.
[0008] Compounds of Formula (I) or (II), as defined herein, may be
used in the treatment of a disease or disorder in which ACMSD plays
a role. The disclosure features methods of treating a disease or
disorder associated with abnormalities in NAD.sup.+ biosynthesis by
administering to subjects suffering from or susceptible to
developing the disease or disorder a therapeutically effective
amount of one or more compounds that increases intracellular
NAD.sup.+ by ACMSD1 inhibition, in an amount sufficient to activate
sirtuins (SIRTs) and the downstream targets of SIRTs, such as
PGC-1.alpha., FoxO1 and/or superoxide dismutase (SOD).
[0009] The methods of the present disclosure can be used in the
treatment of NAD.sup.+ dependent diseases by inhibiting ACMSD.
Inhibition of ACMSD may provide a novel approach to the prevention
and treatment of metabolic disorders, neurodegenerative diseases,
chronic inflammatory diseases, kidney diseases, diseases associated
with ageing and other ACMSD dependent diseases, or diseases
characterized by defective NAD.sup.+ synthesis.
[0010] The present disclosure provides a compound represented by
Formula (I):
##STR00001##
[0011] and pharmaceutically acceptable salts and tautomers thereof,
wherein:
[0012] X is H, S, SR.sup.2, NR.sup.2, NR.sup.2R.sup.2', O, OH,
OR.sup.h, F, Br, or Cl;
[0013] W is N or C;
##STR00002## [0014] (i) when W is N, then: L is
--(C(R.sup.5).sub.2).sub.mCH.dbd.CH(C(R.sup.5).sub.2).sub.p--,
--(C(R.sup.5).sub.2).sub.mY.sup.1(C(R.sup.5).sub.2).sub.p--,
--(C(R.sup.5).sub.2).sub.mY.sup.1(C(R.sup.5).sub.2).sub.p-cyclopropyl-,
--(C(R.sup.5).sub.2).sub.mY.sup.1CH.dbd.CH--,
--(C(R.sup.5).sub.2).sub.mNR.sup.3C.dbd.(O)(C(R.sup.5).sub.2).sub.p--,
--(C(R.sup.5).sub.2).sub.mphenyl(C(R.sup.5).sub.2).sub.p--,
--(C(R.sup.5).sub.2).sub.mpyridinyl(C(R.sup.5).sub.2).sub.p--, or
--(C(R.sup.5).sub.2).sub.mthiophenyl(C(R.sup.5).sub.2).sub.p--;
[0015] (ii) when W is C, then: L is
--(C(R.sup.5).sub.2).sub.mCH.dbd.CH(C(R.sup.5).sub.2).sub.p--,
--(C(R.sup.5).sub.2).sub.o--,
--(C(R.sup.5).sub.2).sub.mY.sup.1(C(R.sup.5).sub.2).sub.p--,
[0015] ##STR00003## --(C(R.sup.5).sub.2).sub.m Y.sup.1CH.dbd.CH--,
--(C(R.sup.5).sub.2).sub.mC.dbd.(O)(CH.sub.2).sub.p,
--(C(R.sup.5).sub.2).sub.mC.dbd.(O)O(C(R.sup.5).sub.2).sub.p--,
--(C(R.sup.5).sub.2).sub.mC.dbd.(O)NR.sup.3(C(R.sup.5).sub.2).sub.p--,
--(C(R.sup.5).sub.2).sub.mNR.sup.3C.dbd.(O)(C(R.sup.5).sub.2).sub.p--,
--(C(R.sup.5).sub.2).sub.mphenyl(C(R.sup.5).sub.2).sub.p--,
--(C(R.sup.5).sub.2).sub.mpyridinyl(C(R.sup.5).sub.2).sub.p--, or
--(C(R.sup.5).sub.2).sub.mthiophenyl(C(R.sup.5).sub.2).sub.p--;
[0016] Y.sup.1 is O, NR.sup.4, or S(O).sub.q;
[0017] each Y.sup.2 is independently O, NH or S;
[0018] R.sup.1 is absent or C.sub.6-C.sub.10 arylene or
heteroarylene, wherein the heteroarylene comprises one or two 5- to
7-membered rings and 1-4 heteroatoms selected from N, O and S, and
wherein the C.sub.6-C.sub.10 arylene or heteroarylene are
optionally substituted with one to two R.sup.e;
[0019] R.sup.2 is H or C.sub.1-C.sub.4 alkyl;
[0020] R.sup.2 is H, C.sub.1-C.sub.4 alkyl, or C.sub.3-C.sub.7
cycloalkyl; or
[0021] R.sup.2 and R.sup.2' together with the nitrogen atom to
which they are attached form a 3- to 7-membered heterocycloalkyl
ring comprising 1-3 additional heteroatoms selected from N, O and
S;
[0022] R.sup.3 is H or C.sub.1-C.sub.4 alkyl;
[0023] R.sup.4 is H or C.sub.1-C.sub.4 alkyl;
[0024] each R.sup.5 is independently at each occurrence H or
C1-C.sub.4 alkyl;
[0025] each R.sup.6 is independently at each occurrence H or
C.sub.1-C.sub.4 alkyl;
[0026] R.sup.7 is H, A, B, or C;
[0027] A is --(C(R.sup.6).sub.2).sub.rCO.sub.2R.sup.x,
--Y.sup.2(C(R.sup.6).sub.2).sub.rCO.sub.2R.sup.x,
--(CH.sub.2).sub.rtetrazole, --(CH.sub.2).sub.roxadiazolone,
--(CH.sub.2).sub.rtetrazolone, --(CH.sub.2).sub.rthiadiazolol,
--(CH.sub.2).sub.r isoxazol-3-ol,
--(CH.sub.2).sub.rP(O)(OH)OR.sup.x, --(CH.sub.2).sub.rS(O).sub.2OH,
--(CH.sub.2).sub.rC(O)NHCN, or
--(CH.sub.2).sub.rC(O)NHS(O).sub.2alkyl, wherein
--(CH.sub.2).sub.rtetrazole, --(CH.sub.2).sub.roxadiazolone,
--(CH.sub.2).sub.rtetrazolone, --(CH.sub.2).sub.rthiadiazolol,
--(CH.sub.2).sub.r isoxazol-3-ol are optionally substituted with
C.sub.1-C.sub.6 alkyl,
[0028] B is --(C(R.sup.6).sub.2).sub.rS(O).sub.2OC.sub.1-C.sub.4
alkyl, --O(C(R.sup.6).sub.2).sub.rS(O).sub.2OC.sub.1-C.sub.4 alkyl,
--Y.sup.2(C(R.sup.6).sub.2).sub.rC(O)NR.sup.gR.sup.g',
--Y.sup.2(C(R.sup.6).sub.2).sub.rS(O).sub.2NR.sup.gR.sup.g',
--(CH.sub.2).sub.rC(O)NR.sup.gR.sup.g',
--(CH.sub.2).sub.rS(O).sub.2NR.sup.gR.sup.g',
--(CH.sub.2).sub.rC(O)NHS(O).sub.2NR.sup.gR.sup.g',
--(C(R.sup.6).sub.2).sub.rCO.sub.2R.sup.i,
--(C(R.sup.6).sub.2).sub.rNH.sub.2CO.sub.2R.sup.x,
--(C(R.sup.6).sub.2).sub.rP(O)(OR.sup.x).sub.2,
--O(C(R.sup.6).sub.2).sub.rP(O)(OR.sup.x).sub.2,
--(C(R.sup.6).sub.2).sub.rS(O).sub.2OH,
--O(C(R.sup.6).sub.2).sub.rS(O).sub.2OH,
--(C(R.sup.6).sub.2).sub.rP(O).sub.2OR.sup.x, or
--O(C(R.sup.6).sub.2).sub.rP(O).sub.2OR.sup.x,
[0029] C is --(CH.sub.2).sub.rCN, --(CH.sub.2).sub.sOH, halogen,
--(C(R.sup.6).sub.2).sub.rC.sub.6-C.sub.10 aryl,
--(C(R.sup.6).sub.2).sub.rS--C.sub.6-C.sub.10 aryl,
--(C(R.sup.6).sub.2).sub.rheteroaryl,
--O(C(R.sup.6).sub.2).sub.rheteroaryl,
--O(C(R.sup.6).sub.2).sub.rheterocycloalkyl,
--O(C(R.sup.6).sub.2).sub.rOH, --OR.sup.y,
--(C(R.sup.6).sub.2).sub.rC(O)NHCN, --CH.dbd.CHCO.sub.2R.sup.x, or
--(C(R.sup.6).sub.2).sub.rC(O)NHS(O).sub.2C.sub.1-C.sub.4 alkyl,
wherein the aryl and heteroaryl are substituted with one to three
substituents each independently selected from C.sub.1-C.sub.6
alkyl, C.sub.1-C.sub.6 haloalkyl, halogen, and OH, and wherein the
heterocycloalkyl is substituted with one to two .dbd.O or
.dbd.S;
[0030] R.sup.c is H, C.sub.1-C.sub.6 alkyl, C.sub.1-C.sub.6
haloalkyl, halogen, --CN, --OR.sup.x, or --CO.sub.2R.sup.x;
[0031] R.sup.d is methyl, CF.sub.3, CR.sup.fF.sub.2,
--(C(R.sup.6).sub.2).sub.tC.sub.6-C.sub.10 aryl,
--(C(R.sup.6).sub.2).sub.t-5- or 6-membered heteroaryl,
--(C(R.sup.6).sub.2).sub.t-5- or 6-membered cycloalkyl, optionally
substituted C.sub.6-C.sub.10 aryl, optionally substituted 5- or
6-membered heteroaryl, or optionally substituted 5- or 6-membered
cycloalkyl;
[0032] each R.sup.e is independently at each occurrence
C.sub.1-C.sub.6 alkyl, C.sub.2-C.sub.6 alkenyl, C.sub.2-C.sub.6
alkynyl, halogen, C.sub.1-C.sub.6 haloalkyl, --NHR.sup.z, --OH, or
--CN;
[0033] R.sup.f is absent, H, or methyl;
[0034] R.sup.g is H, C.sub.1-C.sub.6 alkyl, OH,
--S(O).sub.2(C.sub.1-C.sub.6 alkyl), or S(O).sub.2N(C.sub.1-C.sub.6
alkyl).sub.2;
[0035] R.sup.g' is H, C.sub.1-C.sub.6 alkyl, C.sub.3-C.sub.7
cycloalkyl, 4- to 7-membered heterocycloalkyl ring comprising 1-3
heteroatoms selected from N, O and S, C.sub.6-C.sub.10 aryl, or 5-
to 7-membered heteroaryl comprising 1-3 heteroatoms selected from
N, O and S, wherein the alkyl is optionally substituted with one or
more substituents independently selected from halogen and --OH, and
wherein the cycloalkyl, heterocycloalkyl, aryl and heteroaryl are
optionally substituted with one or more substituents independently
selected from C.sub.1-C.sub.6 alkyl, halogen, and --OH;
[0036] R.sup.h is H, C.sub.1-C.sub.4 alkyl, or 3- to 7-membered
heterocycloalkyl ring comprising 1-3 heteroatoms selected from N, O
and S, wherein the alkyl is optionally substituted with one or more
substituents each independently selected from NH.sub.2,
C.sub.1-C.sub.4 alkylamino, C.sub.1-C.sub.4 dialkylamino, and
C(O)NH.sub.2; and wherein the heterocycloalkyl is optionally
substituted with one or more substituents each independently
selected from C.sub.1-C.sub.6 alkyl and C.sub.1-C.sub.6
haloalkyl;
[0037] R.sup.i is (i) --(CH.sub.2).sub.sOC(O)C.sub.1-C.sub.6 alkyl,
wherein the alkyl is substituted with one or more NH.sub.2; (ii)
(CH.sub.2CH.sub.2O).sub.nCH.sub.2CH.sub.2OH; or (iii)
C.sub.1-C.sub.6 alkyl substituted with one or more substituents
each independently selected from OH and 4- to 7-membered
heterocycloalkyl comprising 1 to 3 heteroatoms selected from O, N,
or S;
[0038] R.sup.i is absent, H, C.sub.1-C.sub.6 alkyl, or --CN;
[0039] each R.sup.x is independently at each occurrence H,
C.sub.1-C.sub.6 alkyl, or C.sub.6-C.sub.10 aryl;
[0040] each R.sup.y and R.sup.z is independently H, C.sub.1-C.sub.6
alkyl, or C.sub.1-C.sub.6 haloalkyl;
[0041] each m, p, q, r, and t is independently 0, 1 or 2;
[0042] n is 0, 1, 2, or 3;
[0043] s is 1 or 2;
[0044] o is 0, 1, 2, 3, or 4; and
[0045] represents a single bond or a double bond; and
[0046] provided that
[0047] when X is O; R.sup.f is H; W is C; R.sup.j is --CN; L is
--SCH.sub.2--; R.sup.1 is phenylene or pyridine; then R.sup.7 is
not --COOH;
[0048] when X is O; R.sup.f is H; W is C; R.sup.j is --CN; L is
--SCH.sub.2--; R.sup.1 is phenylene or pyridine; and R.sup.7 is
tetrazole; then R.sup.c is not H;
[0049] when X is O; R.sup.f is H; W is C; R.sup.j is --CN; L is
--S--C(R.sup.5).sub.2 or --SCH.sub.2CH.sub.2--; R.sup.1 is absent;
then R.sup.7 is not COOH or tetrazole;
[0050] when X is O, R.sup.f is H; W is N; R.sup.i is absent;
R.sup.d is methyl, optionally substituted 5- to 10-membered aryl,
optionally substituted 5- or 6-membered heteroaryl, or optionally
substituted 5- or 6-membered cycloalkyl; L is --SCH.sub.2-- or
--OCH.sub.2--; and R.sup.1 is phenylene; then R.sup.7 is not --COH,
--CH.sub.2COOH,
##STR00004##
and
[0051] when X is O, R.sup.f is H, W is N, R.sup.j is absent, L is
--NHCH.sub.2--, --CH.sub.2NH--, or --NH--C(O)--, and R.sup.1 is
phenylene, then R.sup.d is not phenyl.
[0052] The present disclosure provides a compound represented by
Formula (II):
##STR00005##
[0053] and pharmaceutically acceptable salts and tautomers thereof,
wherein:
[0054] X is H, S, SR.sup.2, NR.sup.2, NR.sup.2R2, O, OH, OR.sup.h,
F, Br, or Cl;
[0055] W is N or C; [0056] (i) when W is N, then: L is
--(C(R.sup.5).sub.2).sub.mCH.dbd.CH(C(R.sup.5).sub.2).sub.p--,
[0056] ##STR00006##
--(C(R.sup.5).sub.2).sub.mY.sup.1(C(R.sup.5).sub.2).sub.p--,
--(C(R.sup.5).sub.2).sub.mY.sup.1(C(R.sup.5).sub.2).sub.pcyclopropyl-,
--(C(R.sup.5).sub.2).sub.mY.sup.1CH.dbd.CH--,
--(C(R.sup.5).sub.2).sub.mNR.sup.3C.dbd.(O)(C(R.sup.5).sub.2).sub.p--,
--(C(R.sup.5).sub.2).sub.mphenyl(C(R.sup.5).sub.2).sub.p--,
--(C(R.sup.5).sub.2).sub.mpyridinyl(C(R.sup.5).sub.2).sub.p--, or
--(C(R.sup.5).sub.2).sub.mthiophenyl(C(R.sup.5).sub.2).sub.p--;
[0057] (ii) when W is C, then: L is
--(C(R.sup.5).sub.2).sub.mCH.dbd.CH(C(R.sup.5).sub.2).sub.p--,
--(C(R.sup.5).sub.2).sub.o--,
--(C(R.sup.5).sub.2).sub.mY.sup.1(C(R.sup.5).sub.2).sub.p--,
[0057] ##STR00007## --(C(R.sup.5).sub.2).sub.m Y.sup.1CH.dbd.CH--,
--(C(R.sup.5).sub.2).sub.mC.dbd.(O)(CH.sub.2).sub.p,
--(C(R.sup.5).sub.2).sub.mC.dbd.(O)O(C(R.sup.5).sub.2).sub.p--,
--(C(R.sup.5).sub.2).sub.mC.dbd.(O)NR.sup.3(C(R.sup.5).sub.2).sub.p--,
--(C(R.sup.5).sub.2).sub.mNR.sup.3C.dbd.(O)(C(R.sup.5).sub.2).sub.p--,
--(C(R.sup.5).sub.2).sub.mphenyl(C(R.sup.5).sub.2).sub.p--,
--(C(R.sup.5).sub.2).sub.mpyridinyl(C(R.sup.5).sub.2).sub.p--, or
--(C(R.sup.5).sub.2).sub.mthiophenyl(C(R.sup.5).sub.2).sub.p--;
[0058] Y.sup.1 is O, NR.sup.4, or S(O).sub.q;
[0059] each Y.sup.2 is independently O, NH or S;
[0060] R.sup.1 is absent, C.sub.6-C.sub.10 arylene, heteroarylene,
or C.sub.3-C.sub.8cycloalkylene, wherein the heteroarylene
comprises one or two 5- to 7-membered rings and 1-4 heteroatoms
selected from N, O and S, and wherein the C.sub.6-C.sub.10 arylene,
heteroarylene, and C.sub.3-C.sub.8cycloalkylene are optionally
substituted with one to two R.sup.e;
[0061] R.sup.2 is H or C.sub.1-C.sub.4 alkyl;
[0062] R.sup.2' is H, C.sub.1-C.sub.4 alkyl, or C.sub.3-C.sub.7
cycloalkyl; or
[0063] R.sup.2 and R.sup.2' together with the nitrogen atom to
which they are attached form a 3- to 7-membered heterocycloalkyl
ring comprising 1-3 additional heteroatoms selected from N, O and
S;
[0064] R.sup.3 is H or C.sub.1-C.sub.4 alkyl;
[0065] R.sup.4 is H or C.sub.1-C.sub.4 alkyl;
[0066] each R.sup.5 is independently at each occurrence H or
C.sub.1-C.sub.4 alkyl;
[0067] each R.sup.6 is independently at each occurrence H or
C.sub.1-C.sub.4 alkyl;
[0068] R.sup.7 is H, A, B, or C;
[0069] A is --(C(R.sup.6).sub.2).sub.rCO.sub.2R.sup.x,
--Y.sup.2(C(R.sup.6).sub.2).sub.rCO.sub.2R.sup.x,
--(C(R.sup.6).sub.2).sub.rtetrazole,
--(C(R.sup.6).sub.2).sub.roxadiazolone,
--(C(R.sup.6).sub.2).sub.rtetrazolone,
--(C(R.sup.6).sub.2).sub.rthiadiazolol, --(C(R.sup.6).sub.2).sub.r
isoxazol-3-ol, --(C(R.sup.6).sub.2).sub.rP(O)(OH)OR.sup.x,
--(C(R.sup.6).sub.2).sub.rS(O).sub.2OH,
--(C(R.sup.6).sub.2).sub.rC(O)NHCN, or
--(C(R.sup.6).sub.2).sub.rC(O)NHS(O).sub.2alkyl, wherein
--(C(R.sup.6).sub.2).sub.rtetrazole,
--(C(R.sup.6).sub.2).sub.roxadiazolone,
--(C(R.sup.6).sub.2).sub.rtetrazolone,
--(C(R.sup.6).sub.2).sub.rthiadiazolol, --(C(R.sup.6).sub.2).sub.r
isoxazol-3-ol are optionally substituted with C.sub.1-C.sub.6
alkyl,
[0070] B is --(C(R.sup.6).sub.2).sub.rS(O).sub.2OC.sub.1-C.sub.4
alkyl, --O(C(R.sup.6).sub.2).sub.rS(O).sub.2OC.sub.1-C.sub.4 alkyl,
--Y.sup.2(C(R.sup.6).sub.2).sub.rC(O)NR.sup.gR.sup.g',
--Y.sup.2(C(R.sup.6).sub.2).sub.rS(O).sub.2NR.sup.gR.sup.g',
--(C(R.sup.6).sub.2).sub.rC(O)NR.sup.gR.sup.g',
--(C(R.sup.6).sub.2).sub.rS(O).sub.2NR.sup.gR.sup.g',
--(C(R.sup.6).sub.2).sub.rC(O)NHS(O).sub.2NR.sup.gR.sup.g',
--(C(R.sup.6).sub.2).sub.rCO.sub.2R.sup.w,
--(C(R.sup.6).sub.2).sub.rNH.sub.2CO.sub.2R.sup.x,
--(C(R.sup.6).sub.2).sub.rP(O)(OR.sup.x).sub.2,
--O(C(R.sup.6).sub.2).sub.rP(O)(OR.sup.x).sub.2,
--(C(R.sup.6).sub.2).sub.rS(O).sub.2OH,
--O(C(R.sup.6).sub.2).sub.rS(O).sub.2OH,
--(C(R.sup.6).sub.2).sub.rP(O).sub.2OR.sup.x, or
--O(C(R.sup.6).sub.2).sub.rP(O).sub.2OR.sup.x,
[0071] C is --(CH.sub.2).sub.rCN, --(CH.sub.2).sub.sOH, halogen,
--(C(R.sup.6).sub.2).sub.rC.sub.6-C.sub.10 aryl,
--(C(R.sup.6).sub.2).sub.rS--C.sub.6-C.sub.10 aryl,
--(C(R.sup.6).sub.2).sub.rheteroaryl,
--O(C(R.sup.6).sub.2).sub.rheteroaryl,
--O(C(R.sup.6).sub.2).sub.rheterocycloalkyl,
--O(C(R.sup.6).sub.2).sub.rOH, --OR.sup.y,
--(C(R.sup.6).sub.2).sub.rC(O)NHCN, --CH.dbd.CHCO.sub.2R.sup.x, or
--(C(R.sup.6).sub.2).sub.rC(O)NHS(O).sub.2C.sub.1-C.sub.4 alkyl,
wherein the aryl and heteroaryl are substituted with one to three
substituents each independently selected from C.sub.1-C.sub.6
alkyl, C.sub.1-C.sub.6 haloalkyl, halogen, and OH, and wherein the
heterocycloalkyl is substituted with one to two .dbd.O or
.dbd.S;
[0072] R.sup.c is H, C.sub.1-C.sub.6 alkyl, C.sub.1-C.sub.6
haloalkyl, halogen, --CN, --OR.sup.x, or --CO.sub.2R.sup.x;
[0073] R.sup.d is methyl, CF.sub.3, CR.sup.fF.sub.2,
--(C(R.sup.6).sub.2).sub.tC.sub.6-C.sub.10 aryl,
--(C(R.sup.6).sub.2).sub.t-5- or 6-membered heteroaryl,
--(C(R.sup.6).sub.2).sub.t-5- or 6-membered cycloalkyl, optionally
substituted C.sub.6-C.sub.10 aryl, optionally substituted 5- or
6-membered heteroaryl, or optionally substituted 5- or 6-membered
cycloalkyl;
[0074] each R.sup.e is independently at each occurrence
C.sub.1-C.sub.6 alkyl, C.sub.2-C.sub.6 alkenyl, C.sub.2-C.sub.6
alkynyl, halogen, C.sub.1-C.sub.6 haloalkyl, --NHR.sup.z, --OH, or
--CN;
[0075] R.sup.f is absent, H, or methyl;
[0076] R.sup.g is H, C.sub.1-C.sub.6 alkyl, OH,
--S(O).sub.2(C.sub.1-C.sub.6 alkyl), or S(O).sub.2N(C.sub.1-C.sub.6
alkyl).sub.2;
[0077] R.sup.g' is H, C.sub.1-C.sub.6 alkyl, C.sub.3-C.sub.7
cycloalkyl, 4- to 7-membered heterocycloalkyl ring comprising 1-3
heteroatoms selected from N, O and S, C.sub.6-C.sub.10 aryl, or 5-
to 7-membered heteroaryl comprising 1-3 heteroatoms selected from
N, O and S, wherein the alkyl is optionally substituted with one or
more substituents independently selected from halogen and --OH, and
wherein the cycloalkyl, heterocycloalkyl, aryl and heteroaryl are
optionally substituted with one or more substituents independently
selected from C.sub.1-C.sub.6 alkyl, halogen, and --OH;
[0078] R.sup.h is H, C.sub.1-C.sub.4 alkyl, or 3- to 7-membered
heterocycloalkyl ring comprising 1-3 heteroatoms selected from N, O
and S, wherein the alkyl is optionally substituted with one or more
substituents each independently selected from NH.sub.2,
C.sub.1-C.sub.4 alkylamino, C.sub.1-C.sub.4 dialkylamino, and
C(O)NH.sub.2; and wherein the heterocycloalkyl is optionally
substituted with one or more substituents each independently
selected from C.sub.1-C.sub.6 alkyl and C.sub.1-C.sub.6
haloalkyl;
[0079] R.sup.i is (i) --(CH.sub.2).sub.sOC(O)C.sub.1-C.sub.6 alkyl,
wherein the alkyl is substituted with one or more NH.sub.2; (ii)
(CH.sub.2CH.sub.2O).sub.nCH.sub.2CH.sub.2OH; or (iii)
C.sub.1-C.sub.6 alkyl substituted with one or more substituents
each independently selected from OH and 4- to 7-membered
heterocycloalkyl comprising 1 to 3 heteroatoms selected from O, N,
or S;
[0080] R.sup.i is absent, H, C.sub.1-C.sub.6 alkyl, or --CN;
[0081] each R.sup.x is independently at each occurrence H,
C.sub.1-C.sub.6 alkyl, or C.sub.6-C.sub.10 aryl;
[0082] each R.sup.y and R.sup.z is independently H, C.sub.1-C.sub.6
alkyl, or C.sub.1-C.sub.6 haloalkyl;
[0083] each m, p, q, r, and t is independently 0, 1 or 2;
[0084] n is 0, 1, 2, or 3;
[0085] s is 1 or 2;
[0086] o is 0, 1, 2, 3, or 4; and
[0087] represents a single bond or a double bond; and
[0088] provided that
[0089] when X is O; R.sup.f is H; W is C; R.sup.j is --CN; L is
--SCH.sub.2--; R.sup.1 is phenylene or pyridine; then R.sup.7 is
not --COOH;
[0090] when X is O; R.sup.f is H; W is C; R.sup.j is --CN; L is
--SCH.sub.2--; R.sup.1 is phenylene or pyridine; and R.sup.7 is
tetrazole; then R.sup.c is not H;
[0091] when X is O; R.sup.f is H; W is C; R.sup.j is --CN; L is
--S--C(R.sup.5).sub.2 or --SCH.sub.2CH.sub.2--; R.sup.1 is absent;
then R.sup.7 is not COOH or tetrazole;
[0092] when X is O, R.sup.f is H; W is N; R.sup.i is absent;
R.sup.d is methyl, optionally substituted 5- to 10-membered aryl,
optionally substituted 5- or 6-membered heteroaryl, or optionally
substituted 5- or 6-membered cycloalkyl; L is --SCH.sub.2-- or
--OCH.sub.2--; and R.sup.1 is phenylene; then R.sup.7 is not
--COOH, --CH.sub.2COOH,
##STR00008##
and
[0093] when X is O, R.sup.f is H, W is N, R.sup.j is absent, L is
--NHCH.sub.2--, --CH.sub.2NH--, or --NH--C(O)--, and R.sup.1 is
phenylene, then R.sup.d is not phenyl.
[0094] Another aspect of the present disclosure provides a
pharmaceutical composition comprising a compound of Formula (I) or
(II), or a pharmaceutically acceptable salt thereof, and at least
one of a pharmaceutically acceptable carrier, diluent, or
excipient.
[0095] Another aspect of the present disclosure provides a compound
of Formula (I) or (II) for use as a medicament. Another aspect of
the present disclosure provides a pharmaceutical composition
comprising a compound of Formula (I) or (II) for use as a
medicament.
[0096] Another aspect of the present disclosure provides a method
of treating a disease or disorder by inhibition of
.alpha.-amino-.beta.-carboxymuconate-.epsilon.-semialdehyde
decarboxylase (ACMSD) comprising administering to the subject
suffering from or susceptible to developing the disease or disorder
a therapeutically effective amount of one or more compounds of
Formula (I) or (II). Another aspect of the present disclosure
provides a method of preventing a disease or disorder by inhibition
of .alpha.-amino-.beta.-carboxymuconate-.epsilon.-semialdehyde
decarboxylase (ACMSD) comprising administering to the subject
suffering from or susceptible to developing the disease or disorder
a therapeutically effective amount of one or more compounds of
Formula (I) or (II). Another aspect of the present disclosure
provides a method of reducing the risk of a disease or disorder by
inhibition of
.alpha.-amino-.beta.-carboxymuconate-.epsilon.-semialdehyde
decarboxylase (ACMSD) comprising administering to the subject
suffering from or susceptible to developing the disease or disorder
a therapeutically effective amount of one or more compounds of
Formula (I) or (II).
[0097] Another aspect of the present disclosure provides a method
of treating a disease or disorder associated with reduced
nicotinamide adenine dinucleotide (NAD.sup.+) levels comprising
administering to the subject suffering from or susceptible to
developing a disease or disorder associated with reduced NAD.sup.+
levels a therapeutically effective amount of one or more compounds
of Formula (I) or (II). Another aspect of the present disclosure
provides a method of preventing a disease or disorder associated
with reduced nicotinamide adenine dinucleotide (NAD.sup.+) levels
comprising administering to the subject suffering from or
susceptible to developing a disease or disorder associated with
reduced NAD.sup.+ levels a therapeutically effective amount of one
or more compounds of Formula (I) or (II). Another aspect of the
present disclosure provides a method of reducing the risk of a
disease or disorder associated with reduced nicotinamide adenine
dinucleotide (NAD.sup.+) levels comprising administering to the
subject suffering from or susceptible to developing a disease or
disorder associated with reduced NAD.sup.+ levels a therapeutically
effective amount of one or more compounds of Formula (I) or
(II).
[0098] Another aspect of the present disclosure provides a method
of treating a disorder associated with mitochondrial dysfunction
comprising administering to the subject suffering from or
susceptible to developing a metabolic disorder a therapeutically
effective amount of one or more compounds of Formula (I) or (II)
that increases intracellular nicotinamide adenine dinucleotide
(NAD.sup.+). Another aspect of the present disclosure provides a
method of preventing a disorder associated with mitochondrial
dysfunction comprising administering to the subject suffering from
or susceptible to developing a metabolic disorder a therapeutically
effective amount of one or more compounds of Formula (I) or (II)
that increases intracellular nicotinamide adenine dinucleotide
(NAD.sup.+). Another aspect of the present disclosure provides a
method of reducing the risk of a disorder associated with
mitochondrial dysfunction comprising administering to the subject
suffering from or susceptible to developing a metabolic disorder a
therapeutically effective amount of one or more compounds of
Formula (I) or (II) that increases intracellular nicotinamide
adenine dinucleotide (NAD.sup.+).
[0099] Another aspect of the present disclosure provides a method
of promoting oxidative metabolism comprising administering to the
subject suffering from or susceptible to developing a metabolic
disorder a therapeutically effective amount of one or more
compounds of Formula (I) or (II) that increases intracellular
nicotinamide adenine dinucleotide (NAD.sup.+).
[0100] Another aspect of the present disclosure provides a compound
of Formula (I) or (II) for use in treating a disease or disorder by
inhibition of
.alpha.-amino-.beta.-carboxymuconate-.epsilon.-semialdehyde
decarboxylase (ACMSD). Another aspect of the present disclosure
provides a compound of Formula (I) or (II) for use in preventing a
disease or disorder by inhibition of
.alpha.-amino-.beta.-carboxymuconate-.epsilon.-semialdehyde
decarboxylase (ACMSD). Another aspect of the present disclosure
provides a compound of Formula (I) or (II) for use in reducing the
risk of a disease or disorder by inhibition of
.alpha.-amino-.beta.-carboxymuconate-.epsilon.-semialdehyde
decarboxylase (ACMSD).
[0101] Another aspect of the present disclosure provides a compound
of Formula (I) or (II) for use in treating a disease or disorder
associated with reduced nicotinamide adenine dinucleotide
(NAD.sup.+) levels. Another aspect of the present disclosure
provides a compound of Formula (I) or (II) for use in preventing a
disease or disorder associated with reduced nicotinamide adenine
dinucleotide (NAD.sup.+) levels. Another aspect of the present
disclosure provides a compound of Formula (I) or (II) for use in
reducing the risk of a disease or disorder associated with reduced
nicotinamide adenine dinucleotide (NAD.sup.+) levels.
[0102] Another aspect of the present disclosure provides a compound
of Formula (I) or (II) that increases intracellular nicotinamide
adenine dinucleotide (NAD.sup.+) for use in treating a disorder
associated with mitochondrial dysfunction. Another aspect of the
present disclosure provides a compound of Formula (I) or (II) that
increases intracellular nicotinamide adenine dinucleotide
(NAD.sup.+) for use in preventing a disorder associated with
mitochondrial dysfunction. Another aspect of the present disclosure
provides a compound of Formula (I) or (II) that increases
intracellular nicotinamide adenine dinucleotide (NAD.sup.+) for use
in reducing the risk of a disorder associated with mitochondrial
dysfunction.
[0103] Another aspect of the present disclosure provides a compound
of Formula (I) or (II) that increases intracellular nicotinamide
adenine dinucleotide (NAD.sup.+) for use in promoting oxidative
metabolism.
[0104] Another aspect of the present disclosure provides use of a
compound of Formula (I) or (II) for treating a disease or disorder
by inhibition of
.alpha.-amino-.beta.-carboxymuconate-.epsilon.-semialdehyde
decarboxylase (ACMSD). Another aspect of the present disclosure
provides use of a compound of Formula (I) or (II) for preventing a
disease or disorder by inhibition of
.alpha.-amino-.beta.-carboxymuconate-.epsilon.-semialdehyde
decarboxylase (ACMSD). Another aspect of the present disclosure
provides use of a compound of Formula (I) or (II) for reducing the
risk of a disease or disorder by inhibition of
.alpha.-amino-.beta.-carboxymuconate-.epsilon.-semialdehyde
decarboxylase (ACMSD).
[0105] Another aspect of the present disclosure provides use of a
compound of Formula (I) or (II) for treating a disease or disorder
associated with reduced nicotinamide adenine dinucleotide
(NAD.sup.+) levels. Another aspect of the present disclosure
provides use of a compound of Formula (I) or (II) for preventing a
disease or disorder associated with reduced nicotinamide adenine
dinucleotide (NAD.sup.+) levels. Another aspect of the present
disclosure provides use of a compound of Formula (I) or (II) for
reducing the risk of a disease or disorder associated with reduced
nicotinamide adenine dinucleotide (NAD.sup.+) levels.
[0106] Another aspect of the present disclosure provides use of a
compound of Formula (I) or (II) that increases intracellular
nicotinamide adenine dinucleotide (NAD.sup.+) for treating a
disorder associated with mitochondrial dysfunction. Another aspect
of the present disclosure provides use of a compound of Formula (I)
or (II) that increases intracellular nicotinamide adenine
dinucleotide (NAD.sup.+) for preventing a disorder associated with
mitochondrial dysfunction. Another aspect of the present disclosure
provides use of a compound of Formula (I) or (II) that increases
intracellular nicotinamide adenine dinucleotide (NAD.sup.+) for
reducing the risk of a disorder associated with mitochondrial
dysfunction.
[0107] Another aspect of the present disclosure provides use of a
compound of Formula (I) or (II) that increases intracellular
nicotinamide adenine dinucleotide (NAD.sup.+) for promoting
oxidative metabolism.
[0108] Another aspect of the present disclosure provides use of a
compound of Formula (I) or (II) in the manufacture of a medicament
for treating a disease or disorder by inhibition of
.alpha.-amino-.beta.-carboxymuconate-.epsilon.-semialdehyde
decarboxylase (ACMSD). Another aspect of the present disclosure
provides use of a compound of Formula (I) or (II) in the
manufacture of a medicament for preventing a disease or disorder by
inhibition of
.alpha.-amino-.beta.-carboxymuconate-.epsilon.-semialdehyde
decarboxylase (ACMSD). Another aspect of the present disclosure
provides use of a compound of Formula (I) or (II) in the
manufacture of a medicament for reducing the risk of a disease or
disorder by inhibition of
.alpha.-amino-.beta.-carboxymuconate-.epsilon.-semialdehyde
decarboxylase (ACMSD).
[0109] Another aspect of the present disclosure provides use of a
compound of Formula (I) or (II) in the manufacture of a medicament
for treating a disease or disorder associated with reduced
nicotinamide adenine dinucleotide (NAD.sup.+) levels. Another
aspect of the present disclosure provides use of a compound of
Formula (I) or (II) in the manufacture of a medicament for
preventing a disease or disorder associated with reduced
nicotinamide adenine dinucleotide (NAD.sup.+) levels. Another
aspect of the present disclosure provides use of a compound of
Formula (I) or (II) in the manufacture of a medicament for reducing
the risk of a disease or disorder associated with reduced
nicotinamide adenine dinucleotide (NAD.sup.+) levels.
[0110] Another aspect of the present disclosure provides use of a
compound of Formula (I) or (II) that increases intracellular
nicotinamide adenine dinucleotide (NAD.sup.+) in the manufacture of
a medicament for treating a disorder associated with mitochondrial
dysfunction. Another aspect of the present disclosure provides use
of a compound of Formula (I) or (II) that increases intracellular
nicotinamide adenine dinucleotide (NAD.sup.+) in the manufacture of
a medicament for preventing a disorder associated with
mitochondrial dysfunction. Another aspect of the present disclosure
provides use of a compound of Formula (I) or (II) that increases
intracellular nicotinamide adenine dinucleotide (NAD.sup.+) in the
manufacture of a medicament for reducing the risk of a disorder
associated with mitochondrial dysfunction.
[0111] Another aspect of the present disclosure provides use of a
compound of Formula (I) or (II) that increases intracellular
nicotinamide adenine dinucleotide (NAD.sup.+) in the manufacture of
a medicament for promoting oxidative metabolism.
[0112] In certain aspects, the compounds of the present disclosure
may be administered alone or in combination with other compounds,
including other ACMSD modulating compounds, or other therapeutic
agents.
[0113] Unless otherwise defined, all technical and scientific terms
used herein have the same meaning as commonly understood by one of
ordinary skill in the art to which this disclosure belongs. In the
specification, the singular forms also include the plural unless
the context clearly dictates otherwise. Although methods and
materials similar to or equivalent to those described herein can be
used in the practice and testing of the disclosure, suitable
methods and materials are described below. All publications, patent
applications, patents, and other references mentioned herein are
incorporated by reference. The references cited herein are not
admitted to be prior art to the claimed disclosure. In the case of
conflict, the present specification, including definitions, will
control. In addition, the materials, methods, and examples are
illustrative only and not intended to be limiting.
[0114] Other features and advantages of the disclosure will be
apparent from the following detailed description and claims.
DETAILED DESCRIPTION OF THE DISCLOSURE
[0115] All references, including any patent or patent application,
cited in this specification are hereby incorporated by reference.
No admission is made that any reference constitutes prior art.
Further, no admission is made that any of the prior art constitutes
part of the common general knowledge in the art.
[0116] As used throughout this disclosure, the following terms,
unless otherwise indicated, shall be understood to have the
following meanings. If a term is missing, the conventional term as
known to one skilled in the art controls.
[0117] As used herein, the terms "including," "containing," and
"comprising" are used in their open, non-limiting sense. Throughout
the description and claims of this specification, the words
"comprise" and "contain" and variations of the words, for example
"comprising" and "comprises", mean "including but not limited to"
and do not exclude other moieties, additives, components, integers
or steps. Throughout the description and claims of this
specification, the singular encompasses the plural unless the
context otherwise requires. In particular, where the indefinite
article is used, the specification is to be understood as
contemplating plurality as well as singularity, unless the context
requires otherwise.
[0118] The articles "a" and "an" as used in this disclosure may
refer to one or more than one (i.e., to at least one) of the
grammatical object of the article. By way of example, "an element"
may mean one element or more than one element.
[0119] The term "and/or" as used in this disclosure may mean either
"and" or "or" unless indicated otherwise.
[0120] To provide a more concise description, some of the
quantitative expressions given herein are not qualified with the
term "about." It is understood that, whether the term "about" is
used explicitly or not, every quantity given herein is meant to
refer to the actual given value, and it is also meant to refer to
the approximation to such given value that would reasonably be
inferred based on the ordinary skill in the art, including
equivalents and approximations due to the experimental and/or
measurement conditions for such given value.
[0121] Whenever a yield is given as a percentage, such yield refers
to a mass of the entity for which the yield is given with respect
to the maximum amount of the same entity that could be obtained
under the particular stoichiometric conditions. Concentrations that
are given as percentages refer to mass ratios, unless indicated
differently.
[0122] The term "alkyl" as used herein refers to a saturated,
straight or branched hydrocarbon chain. The hydrocarbon chain
preferably contains from one to eight carbon atoms
(C.sub.1-8-alkyl), more preferred from one to six carbon atoms
(C.sub.1-6-alkyl), in particular from one to four carbon atoms
(C.sub.1-4-alkyl), including methyl, ethyl, propyl, isopropyl,
butyl, isobutyl, secondary butyl, tertiary butyl, pentyl,
isopentyl, neopentyl, tertiary pentyl, hexyl, isohexyl, heptyl and
octyl. In a preferred embodiment "alkyl" represents a
C.sub.1-4-alkyl group, which may in particular include methyl,
ethyl, propyl, isopropyl, butyl, isobutyl, secondary butyl, and
tertiary butyl. Correspondingly, the term "alkylene" means the
corresponding biradical (-alkyl-).
[0123] The term "cycloalkyl" or "carbocycle" as used herein refers
to a cyclic alkyl group, preferably containing from three to ten
carbon atoms (C.sub.3-10-cycloalkyl or C.sub.3-10-carbocycle), such
as from three to eight carbon atoms (C.sub.3-8-cycloalkyl or
C.sub.3-10-carbocycle), preferably from three to six carbon atoms
(C.sub.3-6-cycloalkyl or C.sub.3-10-carbocycle), including
cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl and
cyclooctyl. Furthermore, the term "cycloalkyl" as used herein may
also include polycyclic groups such as for example
bicyclo[2.2.2]octyl, bicyclo[2.2.1]heptanyl, decalinyl and
adamantyl. Correspondingly, the term "cycloalkylene" means the
corresponding biradical (-cycloalkyl-). Alkyl and cycloalkyl groups
may be optionally substituted with 1-4 substituents. Examples of
substituents on alkyl groups include, but are not limited to,
alkyl, alkenyl, alkynyl, halogen, haloalkyl, alkoxy, heteroaryl,
aryl, carbocyclyl, hydroxyl, carbamoyl, oxo, and --CN.
[0124] The term "alkenyl" as used herein refers to a straight or
branched hydrocarbon chain or cyclic hydrocarbons containing one or
more double bonds, including di-enes, tri-enes and poly-enes.
Typically, the alkenyl group comprises from two to eight carbon
atoms (C.sub.2-8-alkenyl), such as from two to six carbon atoms
(C.sub.2-6-alkenyl), in particular from two to four carbon atoms
(C.sub.2-4-alkenyl), including at least one double bond. Examples
of alkenyl groups include ethenyl; 1- or 2-propenyl; 1-, 2- or
3-butenyl, or 1,3-but-dienyl; 1-, 2-, 3-, 4- or 5-hexenyl, or
1,3-hex-dienyl, or 1,3,5-hex-trienyl; 1-, 2-, 3-, 4-, 5-, 6-, or
7-octenyl, or 1,3-octadienyl, or 1,3,5-octatrienyl, or
1,3,5,7-octatetraenyl, or cyclohexenyl. Correspondingly, the term
"alkenylene" means the corresponding biradical (-alkenyl-). Alkenyl
groups may be optionally substituted with 1-4 substituents.
Examples of substituents on alkenyl groups include, but are not
limited to, alkyl, alkenyl, alkynyl, halogen, haloalkyl, alkoxy,
heteroaryl, aryl, carbocyclyl, hydroxyl, carbamoyl, oxo, and
--CN.
[0125] The term "alkynyl" as used herein refers to a straight or
branched hydrocarbon chain containing one or more triple bonds,
including di-ynes, tri-ynes and poly-ynes.
[0126] Typically, the alkynyl group comprises of from two to eight
carbon atoms (C.sub.2-8-alkynyl), such as from two to six carbon
atoms (C.sub.2-6-alkynyl), in particular from two to four carbon
atoms (C.sub.2-4-alkynyl), including at least one triple bond.
Examples of preferred alkynyl groups include ethynyl; 1- or
2-propynyl; 1-, 2- or 3-butynyl, or 1,3-but-diynyl; 1-, 2-, 3-, 4-
or 5-hexynyl, or 1,3-hex-diynyl, or 1,3,5-hex-triynyl; 1-, 2-, 3-,
4-, 5-, 6-, or 7-octynyl, or 1,3-oct-diynyl, or 1,3,5-oct-triynyl,
or 1,3,5,7-oct-tetraynyl. Correspondingly, the term "alkynylene"
means the corresponding biradical (-alkynyl-). Alkynyl groups may
be optionally substituted with 1-4 substituents. Examples of
substituents on alkynyl groups include, but are not limited to,
alkyl, alkenyl, alkynyl, halogen, haloalkyl, alkoxy, heteroaryl,
aryl, carbocyclyl, hydroxyl, carbamoyl, oxo, and --CN.
[0127] The terms "halo" and "halogen" as used herein refer to
fluoro, chloro, bromo or iodo. Thus a trihalomethyl group
represents, e.g., a trifluoromethyl group, or a trichloromethyl
group. Preferably, the terms "halo" and "halogen" designate fluoro
or chloro.
[0128] The term "haloalkyl" as used herein refers to an alkyl
group, as defined herein, which is substituted one or more times
with one or more halogen. Examples of haloalkyl groups include, but
are not limited to, trifluoromethyl, difluoromethyl,
pentafluoroethyl, trichloromethyl, etc.
[0129] The term "alkoxy" as used herein refers to an "alkyl-O--"
group, wherein alkyl is as defined above.
[0130] The term "hydroxyalkyl" as used herein refers to an alkyl
group (as defined hereinabove), which alkyl group is substituted
one or more times with hydroxy. Examples of hydroxyalkyl groups
include HO--CH.sub.2--, HO--CH.sub.2--CH.sub.2-- and
CH.sub.3--CH(OH)--.
[0131] The term "oxy" as used herein refers to an "--O--"
group.
[0132] The term "oxo" as used herein refers to an ".dbd.O"
group.
[0133] The term "amine" as used herein refers to primary
(R--NH.sub.2, R.noteq.H), secondary ((R).sub.2--NH,
(R).sub.2.noteq.H) and tertiary ((R).sub.3--N, R.noteq.H) amines. A
substituted amine is intended to mean an amine where at least one
of the hydrogen atoms has been replaced by the substituent.
[0134] The term "carbamoyl" as used herein refers to a
"H.sub.2N(C.dbd.O)--" group.
[0135] The term "aryl" as used herein, unless otherwise indicated,
includes carbocyclic aromatic ring systems derived from an aromatic
hydrocarbon by removal of a hydrogen atom.
[0136] Aryl furthermore includes bi-, tri- and polycyclic ring
systems. Examples of preferred aryl moieties include phenyl,
naphthyl, indenyl, indanyl, fluorenyl, biphenyl, indenyl, naphthyl,
anthracenyl, phenanthrenyl, pentalenyl, azulenyl, and biphenylenyl.
Preferred "aryl" is phenyl, naphthyl or indanyl, in particular
phenyl, unless otherwise stated. Any aryl used may be optionally
substituted. Correspondingly, the term "arylene" means the
corresponding biradical (-aryl-). Aryl groups may be optionally
substituted with 1-4 substituents. Examples of substituents on aryl
groups include, but are not limited to, alkyl, alkenyl, alkynyl,
halogen, haloalkyl, alkoxy, heteroaryl, aryl, carbocyclyl,
hydroxyl, and --CN.
[0137] The term "heteroaryl" as used herein, refers to aromatic
groups containing one or more heteroatoms selected from O, S, and
N, preferably from one to four heteroatoms, and more preferably
from one to three heteroatoms. Heteroaryl furthermore includes bi-,
tri- and polycyclic groups, wherein at least one ring of the group
is aromatic, and at least one of the rings contains a heteroatom
selected from O, S, and N. Heteroaryl also include ring systems
substituted with one or more oxo moieties. Examples of preferred
heteroaryl moieties include N-hydroxytetrazolyl,
N-hydroxytriazolyl, N-hydroxyimidazolyl, furanyl, triazolyl,
pyranyl, thiadiazinyl, benzothiophenyl, dihydro-benzo[b]thiophenyl,
xanthenyl, isoindanyl, acridinyl, benzisoxazolyl, quinolinyl,
isoquinolinyl, phteridinyl, azepinyl, diazepinyl, imidazolyl,
thiazolyl, carbazolyl, pyridinyl, pyridazinyl, pyrimidinyl,
pyrazolyl, pyrazinyl, tetrazolyl, furyl, thienyl, isoxazolyl,
oxazolyl, isothiazolyl, pyrrolyl, indolyl, benzimidazolyl,
benzofuranyl, cinnolinyl, indazolyl, indolizinyl, phthalazinyl,
triazinyl, isoindolyl, purinyl, oxadiazolyl, thiadiazolyl,
furazanyl, benzofurazanyl, benzothiophenyl, benzotriazolyl,
benzothiazolyl, benzoxazolyl, quinazolinyl, quinoxalinyl,
naphthyridinyl, dihydroquinolyl, tetrahydroquinolyl,
dihydroisoquinolyl, tetrahydroisoquinolyl, benzofuryl,
furopyridinyl, pyrolopyrimidinyl, azaindolyl, pyrazolinyl,
1,2,4-oxadiazol-5(4H)-one, and pyrazolidinyl.
[0138] Non-limiting examples of partially hydrogenated derivatives
are 1,2,3,4-tetrahydronaphthyl, 1,4-dihydronaphthyl, and 1-octalin.
Correspondingly, the term "heteroarylene" means the corresponding
biradical (-heteroaryl-). Heteroaryl groups may be optionally
substituted with 1-4 substituents. Examples of substituents on
heteroaryl groups include, but are not limited to, alkyl, alkenyl,
alkynyl, halogen, haloalkyl, alkoxy, heteroaryl, aryl, carbocyclyl,
hydroxyl, and --CN.
[0139] The term "heterocyclyl" as used herein, refers to cyclic
non-aromatic groups containing one or more heteroatoms selected
from O, S, and N, preferably from one to four heteroatoms, and more
preferably from one to three heteroatoms. Heterocyclyl furthermore
includes bi-, tri- and polycyclic non-aromatic groups, and at least
one of the rings contains a heteroatom selected from O, S, and N.
Heterocyclyl also include ring systems substituted with one or more
oxo moieties. Examples of heterocyclic groups are oxetane,
pyrrolidinyl, pyrrolyl, 3H-pyrrolyl, oxolanyl, furanyl, thiolanyl,
thiophenyl, pyrazolyl, pyrazolidinyl, imidazolyl, imidazolidinyl,
3H-pyrazolyl, 1,2-oxazolyl, 1,3-oxazolyl, 1,2-thiazolyl,
1,3-thiazolyl, 1,2,5-oxadiazolyl, piperidinyl, pyridinyl, oxanyl,
2-H-pyranyl, 4-H-pyranyl, thianyl, 2H-thiopyranyl, pyridazinyl,
1,2-diazinanyl, pyrimidinyl, 1,3-diazinanyl, pyrazinyl,
piperazinyl, 1,4-dioxinyl, 1,4-dioxanyl, 1,3-diazinanyl,
1,4-oxazinyl, morpholino, thiomorpholino, 1,4-oxathianyl,
benzofuranyl, isobenzofuranyl, indazolyl, benzimidazolyl,
quinolinyl, isoquinolinyl, chromayl, isochromanyl, 4H-chromenyl,
1H-isochromenyl, cinnolinyl, quinazolinyl, quinoxalinyl,
phthalazinyl, purinyl, naphthyridinyl, pteridinyl, indolizinyl,
1H-pyrrolizinyl, 4H-quinolizinyl and aza-8-bicyclo[3.2.1]octane.
Correspondingly, the term "heterocyclylene" means the corresponding
biradical (-heterocyclyl-). Heterocyclyl groups may be optionally
substituted with 1-4 substituents. Examples of substituents on
heterocyclyl groups include, but are not limited, to alkyl,
alkenyl, alkynyl, halogen, haloalkyl, alkoxy, heteroaryl, aryl,
carbocyclyl, hydroxyl, and --CN.
[0140] The term "N-heterocyclic ring" as used herein, refers to a
heterocyclyl or a heteroaryl, as defined hereinabove, having at
least one nitrogen atom, and being bound via a nitrogen atom.
Examples of such N-heterocyclic rings are pyrrolidinyl, pyrrolyl,
3H-pyrrolyl, pyrazolyl, pyrazolidinyl, imidazolyl, imidazolidinyl,
3H-pyrazolyl, 1,2-oxazolyl, 1,2-thiazolyl, 1,3-thiazolyl,
piperidinyl, pyridinyl, pyridazinyl, pyrazinyl, piperazinyl,
morpholino, pyridinyl, pyridazinyl, pyrimidinyl, pyrazolyl,
pyrazinyl, tetrazolyl, etc.
[0141] In the present specification, the structural formula of the
compound represents a certain isomer for convenience in some cases,
but the present disclosure includes all isomers, such as
geometrical isomers, optical isomers based on an asymmetrical
carbon, stereoisomers, tautomers, and the like. Accordingly, it
should be understood that the definition of compounds of Formula
(I) or (II) include each and every individual isomer corresponding
to the Formula: Formula (I) or (II), including cis-trans isomers,
stereoisomers and tautomers, as well as racemic mixtures of these
and pharmaceutically acceptable salts thereof. Hence, the
definition of compounds of Formula (I) or (II) are also intended to
encompass all R- and S-isomers of a chemical structure in any
ratio, e.g., with enrichment (i.e., enantiomeric excess or
diastereomeric excess) of one of the possible isomers and
corresponding smaller ratios of other isomers. In addition, a
crystal polymorphism may be present for the compounds represented
by Formula (I) or (II). It is noted that any crystal form, crystal
form mixture, or anhydride or hydrate thereof is included in the
scope of the present disclosure. Furthermore, so-called metabolite
which is produced by degradation of the present compound in vivo is
included in the scope of the present disclosure.
[0142] "Isomerism" means compounds that have identical molecular
formulae but differ in the sequence of bonding of their atoms or in
the arrangement of their atoms in space. Isomers that differ in the
arrangement of their atoms in space are termed "stereoisomers".
[0143] Stereoisomers that are not mirror images of one another are
termed "diastereoisomers", and stereoisomers that are
non-superimposable mirror images of each other are termed
"enantiomers" or sometimes optical isomers. A mixture containing
equal amounts of individual enantiomeric forms of opposite
chirality is termed a "racemic mixture".
[0144] A carbon atom bonded to four non-identical substituents is
termed a "chiral center".
[0145] "Chiral isomer" means a compound with at least one chiral
center. Compounds with more than one chiral center may exist either
as an individual diastereomer or as a mixture of diastereomers,
termed "diastereomeric mixture". When one chiral center is present,
a stereoisomer may be characterized by the absolute configuration
(R or S) of that chiral center. Absolute configuration refers to
the arrangement in space of the substituents attached to the chiral
center. The substituents attached to the chiral center under
consideration are ranked in accordance with the Sequence Rule of
Cahn, Ingold and Prelog. (Cahn et al., Angew. Chem. Inter. Edit.
1966, 5, 385; errata 511; Cahn et al., Angew. Chem. 1966, 78, 413;
Cahn and Ingold, J. Chem. Soc. 1951 (London), 612; Cahn et al.,
Experientia 1956, 12, 81; Cahn, J. Chem. Educ. 1964, 41, 116).
[0146] Diastereoisomers, i.e., non-superimposable stereochemical
isomers, can be separated by conventional means such as
chromatography, distillation, crystallization or sublimation. The
optical isomers can be obtained by resolution of the racemic
mixtures according to conventional processes, for example by
formation of diastereoisomeric salts by treatment with an optically
active acid or base. Examples of appropriate acids include, without
limitation, tartaric, diacetyltartaric, dibenzoyltartaric,
ditoluoyltartaric and camphorsulfonic acid. The mixture of
diastereomers can be separated by crystallization followed by
liberation of the optically active bases from these salts. An
alternative process for separation of optical isomers includes the
use of a chiral chromatography column optimally chosen to maximize
the separation of the enantiomers. Still another available method
involves synthesis of covalent diastereoisomeric molecules by
reacting compounds of Formula (I) or (II) with an optically pure
acid in an activated form or an optically pure isocyanate. The
synthesized diastereoisomers can be separated by conventional means
such as chromatography, distillation, crystallization or
sublimation, and then hydrolyzed to obtain the enantiomerically
pure compound. The optically active compounds of Formulae (I) can
likewise be obtained by utilizing optically active starting
materials and/or by utilizing a chiral catalyst. These isomers may
be in the form of a free acid, a free base, an ester or a salt.
Examples of chiral separation techniques are given in Chiral
Separation Techniques, A Practical Approach, 2.sup.nd ed. by G.
Subramanian, Wiley-VCH, 2001.
[0147] "Geometric isomer" means the diastereomers that owe their
existence to hindered rotation about double bonds. These
configurations are differentiated in their names by the prefixes
cis and trans, or Z and E, which indicate that the groups are on
the same or opposite side of the double bond in the molecule
according to the Cahn-Ingold-Prelog rules.
[0148] Furthermore, the structures and other compounds discussed in
this disclosure include all atropic isomers thereof. "Atropic
isomers" are a type of stereoisomer in which the atoms of two
isomers are arranged differently in space. Atropic isomers owe
their existence to a restricted rotation caused by hindrance of
rotation of large groups about a central bond. Such atropic isomers
typically exist as a mixture, however as a result of recent
advances in chromatography techniques; it has been possible to
separate mixtures of two atropic isomers in select cases.
[0149] "Tautomer" is one of two or more structural isomers that
exist in equilibrium and is readily converted from one isomeric
form to another. This conversion results in the formal migration of
a hydrogen atom accompanied by a switch of adjacent conjugated
double bonds. Tautomers exist as a mixture of a tautomeric set in
solution. In solid form, usually one tautomer predominates. In
solutions where tautomerization is possible, a chemical equilibrium
of the tautomers will be reached. The exact ratio of the tautomers
depends on several factors, including temperature, solvent and pH.
The concept of tautomers that are interconvertable by
tautomerizations is called tautomerism.
[0150] Of the various types of tautomerism that are possible, two
are commonly observed. In keto-enol tautomerism a simultaneous
shift of electrons and a hydrogen atom occurs. Ring-chain
tautomerism arises as a result of the aldehyde group (--CHO) in a
sugar chain molecule reacting with one of the hydroxy groups (--OH)
in the same molecule to give it a cyclic (ring-shaped) form as
exhibited by glucose.
[0151] Common tautomeric pairs are: ketone-enol, amide-nitrile,
lactam-lactim, amide-imidic acid tautomerism in heterocyclic rings
(e.g., in nucleobases such as guanine, thymine and cytosine),
amine-enamine and enamine-enamine. It is to be understood that the
compounds of the present disclosure may be depicted as different
tautomers. It should also be understood that when compounds have
tautomeric forms, all tautomeric forms are intended to be included
in the scope of the present disclosure, and the naming of the
compounds does not exclude any tautomer form.
[0152] The term "crystal polymorphs", "polymorphs" or "crystal
forms" means crystal structures in which a compound (or a salt or
solvate thereof) can crystallize in different crystal packing
arrangements, all of which have the same elemental composition.
Different crystal forms usually have different X-ray diffraction
patterns, infrared spectral, melting points, density hardness,
crystal shape, optical and electrical properties, stability and
solubility. Recrystallization solvent, rate of crystallization,
storage temperature, and other factors may cause one crystal form
to dominate. Crystal polymorphs of the compounds can be prepared by
crystallization under different conditions.
[0153] Additionally, the compounds of the present disclosure, for
example, the salts of the compounds, can exist in either hydrated
or unhydrated (the anhydrous) form or as solvates with other
solvent molecules. Nonlimiting examples of hydrates include
monohydrates, dihydrates, etc. Nonlimiting examples of solvates
include ethanol solvates, acetone solvates, etc.
[0154] "Solvate" means solvent addition forms that contain either
stoichiometric or non-stoichiometric amounts of solvent. Some
compounds have a tendency to trap a fixed molar ratio of solvent
molecules in the crystalline solid state, thus forming a solvate.
If the solvent is water the solvate formed is a hydrate; and if the
solvent is alcohol, the solvate formed is an alcoholate. Hydrates
are formed by the combination of one or more molecules of water
with one molecule of the substance in which the water retains its
molecular state as H.sub.2O.
[0155] As used herein, a "subject" or "subject in need thereof" is
a subject having a disease or disorder associated with
.alpha.-amino-.beta.-carboxymuconate-.epsilon.-semialdehyde
decarboxylase (ACMSD) dysfunction or inhibited by
.alpha.-amino-.beta.-carboxymuconate-.epsilon.-semialdehyde
decarboxylase (ACMSD). A "subject" includes a mammal. The mammal
can be e.g., any mammal, e.g., a human, primate, bird, mouse, rat,
fowl, dog, cat, cow, horse, goat, camel, sheep or a pig.
Preferably, the mammal is a human.
[0156] The present disclosure is intended to include all isotopes
of atoms occurring in the present compounds. Isotopes include those
atoms having the same atomic number but different mass numbers. By
way of general example and without limitation, isotopes of hydrogen
include tritium and deuterium, and isotopes of carbon include C-13
and C-14.
Compounds
[0157] The present disclosure relates to compounds of Formula
(I):
##STR00009##
[0158] and pharmaceutically acceptable salts and tautomers thereof,
wherein the substituents are as described herein.
[0159] The present disclosure relates to compounds of Formula
(II):
##STR00010##
[0160] and pharmaceutically acceptable salts and tautomers thereof,
wherein the substituents are as described herein.
[0161] In certain embodiments of Formula (I) or (II), wherein W is
N, the present disclosure relates to compounds of Formula
(I-I):
##STR00011##
and pharmaceutically acceptable salts and tautomers thereof,
wherein the substituents are as described herein for Formula (I)
and (II).
[0162] In certain embodiments of Formula (I) or (II), wherein W is
C, the present disclosure relates to compounds of Formula
(I-2):
##STR00012##
[0163] and pharmaceutically acceptable salts and tautomers thereof,
wherein the substituents are as described herein for Formula (I)
and (II).
[0164] In certain embodiments of Formula (I) or (II), wherein
R.sup.1 is phenyl, the present disclosure relates to compounds of
Formula (I-3):
##STR00013##
[0165] and pharmaceutically acceptable salts and tautomers thereof,
wherein the substituents are as described herein for Formula (I)
and (II).
[0166] In certain embodiments of Formula (I) or (II), wherein
R.sup.1 is absent, the present disclosure relates to compounds of
Formula (I-4):
##STR00014##
[0167] and pharmaceutically acceptable salts and tautomers thereof,
wherein the substituents are as described herein for Formula (I)
and (II).
[0168] As described above, X is H, S, SR.sup.2, NR.sup.2,
NR.sup.2R.sup.2', O, OH, OR.sup.h, F, Br, or Cl. In certain
embodiments, X is O, OH, OR.sup.h, F, Br, or Cl. In certain
embodiments, X is H, S, SR.sup.2, NR.sup.2, or NR.sup.2R.sup.2'. In
certain embodiments, X is H. In certain embodiments, X is S. In
certain embodiments, X is SR.sup.2. In certain embodiments, X is
NR.sup.2. In certain embodiments, X is NR.sup.2R.sup.2'. In certain
embodiments, X is O. In certain embodiments, X is OH. In certain
embodiments, X is OR.sup.h. In certain embodiments, X is F. In
certain embodiments, X is Br. In certain embodiments, X is Cl.
[0169] As described above, R.sup.2 is H or C.sub.1-C.sub.4 alkyl.
In certain embodiments, R.sup.2 is H. In certain embodiments,
R.sup.2 is C.sub.1-C.sub.4 alkyl. In certain embodiments, R.sup.2
is --CH.sub.3.
[0170] As described above, R.sup.2' is H, C.sub.1-C.sub.4 alkyl, or
C.sub.3-C.sub.7 cycloalkyl. In certain embodiments, R.sup.2' is H.
In certain embodiments, R.sup.2' is C.sub.1-C.sub.4 alkyl. In
certain embodiments, R.sup.2' is C.sub.3-C.sub.7 cycloalkyl.
[0171] As described above, R.sup.2 and R.sup.2' together with the
nitrogen atom to which they are attached form a 3- to 7-membered
heterocycloalkyl ring comprising 1-3 additional heteroatoms
selected from N, O and S. In certain embodiments, R.sup.2 and
R.sup.2' together with the nitrogen atom to which they are attached
form a 6-membered heterocycloalkyl ring.
[0172] As described above, R.sup.h is H, C.sub.1-C.sub.4 alkyl, or
3- to 7-membered heterocycloalkyl ring comprising 1-3 heteroatoms
selected from N, O and S, wherein the alkyl is optionally
substituted with one or more substituents each independently
selected from NH.sub.2, C.sub.1-C.sub.4 alkylamino, C.sub.1-C.sub.4
dialkylamino, and C(O)NH.sub.2; and wherein the heterocycloalkyl is
optionally substituted with one or more substituents each
independently selected from C.sub.1-C.sub.6 alkyl and
C.sub.1-C.sub.6 haloalkyl. In certain embodiments, R.sup.h is H. In
certain embodiments, R.sup.h is C.sub.1-C.sub.4 alkyl, wherein the
alkyl is optionally substituted with one or more substituents each
independently selected from NH.sub.2, C.sub.1-C.sub.4 alkylamino,
C.sub.1-C.sub.4 dialkylamino, and C(O)NH.sub.2. In certain
embodiments, R.sup.h is 3- to 7-membered heterocycloalkyl ring
comprising 1-3 heteroatoms selected from N, O and S, wherein the
heterocycloalkyl is optionally substituted with one or more
substituents each independently selected from C.sub.1-C.sub.6 alkyl
and C.sub.1-C.sub.6 haloalkyl.
[0173] As described above, R.sup.f is absent, H, or methyl. In
certain embodiments, R.sup.f is absent. In certain embodiments,
R.sup.f is H. In certain embodiments, R.sup.f is methyl.
[0174] As described above, W is N or C. In certain embodiments, W
is N. In certain embodiments, W is C.
[0175] As described above, R.sup.i is absent, H, C.sub.1-C.sub.6
alkyl, or --CN. In certain embodiments, R.sup.i is absent. In
certain embodiments, R.sup.i is H. In certain embodiments, R.sup.i
is C.sub.1-C.sub.6 alkyl. In certain embodiments, R.sup.i is
--CN.
[0176] In certain embodiments, W is N and R.sup.i is absent. In
certain embodiments, W is C and R.sup.i is H, C.sub.1-C.sub.6
alkyl, or --CN. In certain embodiments, W is C and R.sup.i is
--CN.
[0177] As described above, R.sup.c is H, C.sub.1-C.sub.6 alkyl,
C.sub.1-C.sub.6 haloalkyl, halogen, --CN, --OR.sup.x, or
--CO.sub.2R.sup.x. In certain embodiments, R.sup.c is H. In certain
embodiments, R.sup.c is C.sub.1-C.sub.6 alkyl. In certain
embodiments, R.sup.c is C.sub.1-C.sub.6 haloalkyl. In certain
embodiments, R.sup.c is halogen. In certain embodiments, R.sup.c is
--CN. In certain embodiments, R.sup.c is --OR.sup.x. In certain
embodiments, R.sup.c is --CO.sub.2R.sup.x.
[0178] As described above, R.sup.x is independently at each
occurrence H, C.sub.1-C.sub.6 alkyl, or C.sub.6-C.sub.10 aryl. In
certain embodiments, R.sup.x is H. In certain embodiments, R.sup.x
is C.sub.1-C.sub.6 alkyl. In certain embodiments, R.sup.x is
C.sub.6-C.sub.10 aryl.
[0179] As described above, R.sup.d is methyl, CF.sub.3,
CR.sup.fF.sub.2, --(C(R.sup.6).sub.2).sub.tC.sub.6-C.sub.10 aryl,
--(C(R.sup.6).sub.2).sub.t-5- or 6-membered heteroaryl,
--(C(R.sup.6).sub.2).sub.t-5- or 6-membered cycloalkyl, optionally
substituted C.sub.6-C.sub.10 aryl, optionally substituted 5- or
6-membered heteroaryl, or optionally substituted 5- or 6-membered
cycloalkyl.
[0180] In certain embodiments, R.sup.d is methyl. In certain
embodiments, R.sup.d is CF.sub.3. In certain embodiments, R.sup.d
is CR.sup.fF.sub.2. In certain embodiments, R.sup.d is
--(C(R.sup.6).sub.2).sub.tC.sub.6-C.sub.10 aryl. In certain
embodiments, R.sup.d is --CH.sub.2C.sub.6-C.sub.10 aryl. In certain
embodiments, R.sup.d is --CH.sub.2C.sub.6aryl. In certain
embodiments, R.sup.d is --(C(R.sup.6).sub.2).sub.t-5- or 6-membered
heteroaryl. In certain embodiments, R.sup.d is
--(C(R.sup.6).sub.2).sub.t-5- or 6-membered cycloalkyl. In certain
embodiments, R.sup.d is optionally substituted C.sub.6-C.sub.10
aryl. In certain embodiments, R.sup.d is optionally substituted 5-
or 6-membered heteroaryl. In certain embodiments, R.sup.d is
optionally substituted 5- or 6-membered cycloalkyl.
[0181] As described above, R.sup.f is absent, H, or methyl. In
certain embodiments, R.sup.f is absent. In certain embodiments,
R.sup.f is H. In certain embodiments, R.sup.f is methyl.
[0182] As described above, t is 0, 1, or 2. In certain embodiments,
t is 0. In certain embodiments, t is 1. In certain embodiments, t
is 2.
[0183] As described above, when W is N, then L is
--(C(R.sup.5).sub.2).sub.mCH.dbd.CH(C(R.sup.5).sub.2).sub.p--,
##STR00015##
--(C(R.sup.5).sub.2).sub.mY.sup.1(C(R.sup.5).sub.2).sub.p--,
--(C(R.sup.5).sub.2).sub.mY.sup.1(C(R.sup.5).sub.2).sub.pcyclopropyl-,
--(C(R.sup.5).sub.2).sub.mY.sup.1CH.dbd.CH--,
--(C(R.sup.5).sub.2).sub.mNR.sup.3C.dbd.(O)(C(R.sup.5).sub.2).sub.p--,
--(C(R.sup.5).sub.2).sub.mphenyl(C(R.sup.5).sub.2).sub.p--,
--(C(R.sup.5).sub.2).sub.mpyridinyl(C(R.sup.5).sub.2).sub.p--, or
--(C(R.sup.5).sub.2).sub.mthiophenyl(C(R.sup.5).sub.2).sub.p--.
[0184] In certain embodiments, W is N and L is
--(C(R.sup.5).sub.2).sub.mCH.dbd.CH(C(R.sup.5).sub.2).sub.p--. In
certain embodiments, W is N and L is
##STR00016##
In certain embodiments, W is N and L is
--(C(R.sup.5).sub.2).sub.mY.sup.1(C(R.sup.5).sub.2).sub.p--. In
certain embodiments, W is N and L is
--(C(R.sup.5).sub.2).sub.mY.sup.1(C(R.sup.5).sub.2).sub.p
cyclopropyl-. In certain embodiments, W is N and L is
--(C(R.sup.5).sub.2).sub.mY.sup.1CH.dbd.CH--. In certain
embodiments, W is N and L is
--(C(R.sup.5).sub.2).sub.mNR.sup.3C.dbd.(O)(C(R.sup.5).sub.2).sub.p--.
In certain embodiments, W is N and L is
--(C(R.sup.5).sub.2).sub.mphenyl(C(R.sup.5).sub.2).sub.p--. In
certain embodiments, W is N and L is
--(C(R.sup.5).sub.2).sub.mpyridinyl(C(R.sup.5).sub.2).sub.p--. In
certain embodiments, W is N and L is
--(C(R.sup.5).sub.2).sub.mthiophenyl(C(R.sup.5).sub.2).sub.p--. In
certain embodiments, W is N and L is
--(C(R.sup.5).sub.2).sub.mY.sup.1(C(R.sup.5).sub.2).sub.p, such as
--SCH.sub.2-- or --NHCH.sub.2--. In certain embodiments, W is N and
L is --SCH.sub.2--. In certain embodiments, W is N and L is
--NHCH.sub.2--.
[0185] As described above, when W is C, L is
--(C(R.sup.5).sub.2).sub.mCH.dbd.CH(C(R.sup.5).sub.2).sub.p--,
--(C(R.sup.5).sub.2).sub.o--,
--(C(R.sup.5).sub.2).sub.mC.dbd.(O)O(C(R.sup.5).sub.2).sub.p--,
##STR00017##
--(C(R.sup.5).sub.2).sub.mC.dbd.(O)NR.sup.3(C(R.sup.5).sub.2).sub.p--,
--(C(R.sup.5).sub.2).sub.mNR.sup.3C.dbd.(O)(C(R.sup.5).sub.2).sub.p--,
--(C(R.sup.5).sub.2).sub.mphenyl(C(R.sup.5).sub.2).sub.p--,
--(C(R.sup.5).sub.2).sub.mpyridinyl(C(R.sup.5).sub.2).sub.p--, or
--(C(R.sup.5).sub.2).sub.mthiophenyl(C(R.sup.5).sub.2).sub.p--.
[0186] In certain embodiments, W is C and L is
--(C(R.sup.5).sub.2).sub.mCH.dbd.CH(C(R.sup.5).sub.2).sub.p--. In
certain embodiments, W is C and L is --(C(R.sup.5).sub.2).sub.o--.
In certain embodiments, W is C and L is
--(C(R.sup.5).sub.2).sub.mY.sup.1(C(R.sup.5).sub.2).sub.p--. In
certain embodiments, W is C and L is
##STR00018##
In certain embodiments, W is C and L is --(C(R.sup.5).sub.2).sub.m
Y.sup.1CH.dbd.CH--. In certain embodiments, W is C and L is
--(C(R.sup.5).sub.2).sub.mC.dbd.(O)(CH.sub.2).sub.p--. In certain
embodiments, W is C and L is
--(C(R.sup.5).sub.2).sub.mC.dbd.(O)O(C(R.sup.5).sub.2).sub.p--. In
certain embodiments, W is C and L is
--(C(R.sup.5).sub.2).sub.mC.dbd.(O)NR.sup.3(C(R.sup.5).sub.2).sub.p--.
In certain embodiments, W is C and L is
--(C(R.sup.5).sub.2).sub.mNR.sup.3C.dbd.(O)(C(R.sup.5).sub.2).sub.p--.
In certain embodiments, W is C and L is
--(C(R.sup.5).sub.2).sub.mphenyl(C(R.sup.5).sub.2).sub.p--. In
certain embodiments, W is C and L is
--(C(R.sup.5).sub.2).sub.mpyridinyl(C(R.sup.5).sub.2).sub.p--. In
certain embodiments, W is C and L is
--(C(R.sup.5).sub.2).sub.mthiophenyl(C(R.sup.5).sub.2).sub.p--.
[0187] As described above, Y.sup.1 is O, NR.sup.4, or S(O).sub.q.
In certain embodiments, Y.sup.1 is O. In certain embodiments,
Y.sup.1 is NR.sup.4. As described above, R.sup.4 is H or
C.sub.1-C.sub.4 alkyl. In certain embodiments, R.sup.4 is H. In
certain embodiments, R.sup.4 is C.sub.1-C.sub.4 alkyl.
[0188] In certain embodiments, Y.sup.1 is S(O).sub.q. As described
above, q is 0, 1, or 2. In certain embodiments, q is 0. In certain
embodiments, Y.sup.1 is S. In certain embodiments, q is 1. In
certain embodiments, q is 2.
[0189] As described above, each R.sup.5 is independently at each
occurrence H or C.sub.1-C.sub.4 alkyl.
[0190] In certain embodiments, R.sup.5 is H. In certain
embodiments, R.sup.5 is C.sub.1-C.sub.4 alkyl.
[0191] As described above, R.sup.3 is H or C.sub.1-C.sub.4 alkyl.
In certain embodiments, R.sup.3 is H. In certain embodiments,
R.sup.3 is C.sub.1-C.sub.4 alkyl.
[0192] As described above, each m and p is independently 0, 1 or 2.
In certain embodiments, m is 0. In certain embodiments, m is 1. In
certain embodiments, m is 2. In certain embodiments, p is 0. In
certain embodiments, p is 1. In certain embodiments, p is 2.
[0193] As described above, o is 0, 1, 2, 3, or 4. In certain
embodiments, o is 0. In certain embodiments, o is 1. In certain
embodiments, o is 2. In certain embodiments, o is 3. In certain
embodiments, o is 4.
[0194] As described above, R.sup.1 is absent or C.sub.6-C.sub.10
arylene or heteroarylene, wherein the heteroarylene comprises one
or two 5- to 7-membered rings and 1-4 heteroatoms selected from N,
O and S, and wherein the C.sub.6-C.sub.10 arylene or heteroarylene
are optionally substituted with one to two R.sup.e. In certain
embodiments, R.sup.1 is absent. In certain embodiments, R.sup.1 is
C.sub.6-C.sub.10 arylene, which is optionally substituted with one
to two R.sup.e. In certain embodiments, R.sup.1 is heteroarylene,
wherein the heteroarylene comprises one or two 5- to 7-membered
rings and 1-4 heteroatoms selected from N, O and S, and optionally
substituted with one to two R. In certain embodiments of Formula
(II), R.sup.1 is C.sub.3-C.sub.8cycloalkylene, such as
C.sub.3cycloalkylene, C.sub.4cycloalkylene, C.sub.5cycloalkylene,
C.sub.6cycloalkylene, C.sub.7cycloalkylene, or
C.sub.5cycloalkylene
[0195] As described above, each R.sup.e is independently at each
occurrence C.sub.1-C.sub.6 alkyl, C.sub.2-C.sub.6 alkenyl,
C.sub.2-C.sub.6 alkynyl, halogen, C.sub.1-C.sub.6 haloalkyl,
--NHR.sup.z, --OH, or --CN.
[0196] As described above, R.sup.7 is H, A, B, or C. In certain
embodiments, R.sup.7 is H. In certain embodiments, R.sup.7 is A. In
certain embodiments, R.sup.7 is B. In certain embodiments, R.sup.7
is C.
[0197] As described above for Formula (I), A is
--(C(R.sup.6).sub.2).sub.rCO.sub.2R.sup.x,
--Y.sup.2(C(R.sup.6).sub.2).sub.rCO.sub.2R.sup.x,
--(CH.sub.2).sub.rtetrazole, --(CH.sub.2).sub.roxadiazolone,
--(CH.sub.2).sub.rtetrazolone, --(CH.sub.2).sub.rthiadiazolol,
--(CH.sub.2).sub.risoxazol-3-ol,
--(CH.sub.2).sub.rP(O)(OH)OR.sup.x, --(CH.sub.2).sub.rS(O).sub.2OH,
--(CH.sub.2).sub.rC(O)NHCN, or
--(CH.sub.2).sub.rC(O)NHS(O).sub.2alkyl, wherein
--(CH.sub.2).sub.rtetrazole, --(CH.sub.2).sub.roxadiazolone,
--(CH.sub.2).sub.rtetrazolone, --(CH.sub.2).sub.rthiadiazolol,
--(CH.sub.2).sub.r isoxazol-3-ol are optionally substituted with
C.sub.1-C.sub.6 alkyl.
[0198] As described above for Formula (II), A is
--(C(R.sup.6).sub.2).sub.rCO.sub.2R.sup.x,
--Y.sup.2(C(R.sup.6).sub.2).sub.rCO.sub.2R.sup.x,
--(C(R.sup.6).sub.2).sub.rtetrazole,
--(C(R.sup.6).sub.2).sub.roxadiazolone,
--(C(R.sup.6).sub.2).sub.rtetrazolone,
--(C(R.sup.6).sub.2).sub.rthiadiazolol, --(C(R.sup.6).sub.2).sub.r
isoxazol-3-ol, --(C(R.sup.6).sub.2).sub.rP(O)(OH)OR.sup.x,
--(C(R.sup.6).sub.2).sub.rS(O).sub.2OH,
--(C(R.sup.6).sub.2).sub.rC(O)NHCN, or
--(C(R.sup.6).sub.2).sub.rC(O)NHS(O).sub.2alkyl, wherein
--(C(R.sup.6).sub.2).sub.rtetrazole,
--(C(R.sup.6).sub.2).sub.roxadiazolone,
--(C(R.sup.6).sub.2).sub.rtetrazolone,
--(C(R.sup.6).sub.2).sub.rthiadiazolol, --(C(R.sup.6).sub.2).sub.r
isoxazol-3-ol are optionally substituted with C.sub.1-C.sub.6
alkyl. In certain embodiments, A is
--(C(R.sup.6).sub.2).sub.rtetrazole. In certain embodiments, A is
--(C(R.sup.6).sub.2).sub.roxadiazolone. In certain embodiments, A
is --(C(R.sup.6).sub.2).sub.rtetrazolone. In certain embodiments, A
is --(C(R.sup.6).sub.2).sub.rthiadiazolol. In certain embodiments,
A is --(C(R.sup.6).sub.2).sub.r isoxazol-3-ol. In certain
embodiments, A is --(C(R.sup.6).sub.2).sub.rP(O)(OH)OR.sup.x. In
certain embodiments, A is --(C(R.sup.6).sub.2).sub.rS(O).sub.2OH.
In certain embodiments, A is --(C(R.sup.6).sub.2).sub.rC(O)NHCN. In
certain embodiments, A is
--(C(R.sup.6).sub.2).sub.rC(O)NHS(O).sub.2alkyl.
[0199] In certain embodiments, A is
--(C(R.sup.6).sub.2).sub.rCO.sub.2R.sup.x. In certain embodiments,
A is --Y.sup.2(C(R.sup.6).sub.2).sub.rCO.sub.2R.sup.x. In certain
embodiments, A is --(CH.sub.2).sub.rtetrazole. In certain
embodiments, A is --(CH.sub.2).sub.roxadiazolone. In certain
embodiments, A is --(CH.sub.2).sub.rtetrazolone. In certain
embodiments, A is --(CH.sub.2).sub.rthiadiazolol. In certain
embodiments, A is --(CH.sub.2).sub.r isoxazol-3-ol.
[0200] In certain embodiments, A is
--(CH.sub.2).sub.rP(O)(OH)OR.sup.x. In certain embodiments, A is
--(CH.sub.2).sub.rS(O).sub.2OH. In certain embodiments, A is
--(CH.sub.2).sub.rC(O)NHCN. In certain embodiments, A is
--(CH.sub.2).sub.rC(O)NHS(O).sub.2alkyl. In certain embodiments,
--(CH.sub.2).sub.rtetrazole, --(CH.sub.2).sub.roxadiazolone,
--(CH.sub.2).sub.rtetrazolone, --(CH.sub.2).sub.rthiadiazolol,
--(CH.sub.2).sub.r isoxazol-3-ol are optionally substituted with
C.sub.1-C.sub.6 alkyl. In certain embodiments, A is
--(C(R.sup.6).sub.2).sub.rCO.sub.2R.sup.x or
--(CH.sub.2).sub.rtetrazole, wherein --(CH.sub.2).sub.rtetrazole is
optionally substituted with C.sub.1-C.sub.6 alkyl. In certain
embodiments, A is --(C(R.sup.6).sub.2).sub.rCOOH or
--(CH.sub.2).sub.rtetrazole, wherein --(CH.sub.2).sub.rtetrazole is
optionally substituted with C.sub.1-C.sub.6 alkyl. In certain
embodiments, A is --COOH, --CH.sub.2COOH, -tetrazole, or
--(CH.sub.2)tetrazole, wherein tetrazole and
--(CH.sub.2).sub.rtetrazole are optionally substituted with
C.sub.1-C.sub.6 alkyl.
[0201] As described above for Formula (I), B is
--(C(R.sup.6).sub.2).sub.rS(O).sub.2OC.sub.1-C.sub.4 alkyl,
--O(C(R.sup.6).sub.2).sub.rS(O).sub.2OC.sub.1-C.sub.4 alkyl,
--Y.sup.2(C(R.sup.6).sub.2).sub.rC(O)NR.sup.gR.sup.g',
--Y.sup.2(C(R.sup.6).sub.2).sub.rS(O).sub.2NR.sup.gR.sup.g',
--(CH.sub.2).sub.rC(O)NR.sup.gR.sup.g',
--(CH.sub.2).sub.rS(O).sub.2NR.sup.gR.sup.g',
--(CH.sub.2).sub.rC(O)NHS(O).sub.2NR.sup.gR.sup.g',
--(C(R.sup.6).sub.2).sub.rCO.sub.2R.sup.i,
--(C(R.sup.6).sub.2).sub.rNH.sub.2CO.sub.2R.sup.x,
--(C(R.sup.6).sub.2).sub.rP(O)(OR.sup.x).sub.2,
--O(C(R.sup.6).sub.2).sub.rP(O)(OR.sup.x).sub.2,
--(C(R.sup.6).sub.2).sub.rS(O).sub.2OH,
--O(C(R.sup.6).sub.2).sub.rS(O).sub.2OH,
--(C(R.sup.6).sub.2).sub.rP(O).sub.2OR.sup.x, or
--O(C(R.sup.6).sub.2).sub.rP(O).sub.2OR.sup.x.
[0202] As described above for Formula (II), is
--(C(R.sup.6).sub.2).sub.rS(O).sub.2OC.sub.1-C.sub.4 alkyl,
--O(C(R.sup.6).sub.2).sub.rS(O).sub.2OC.sub.1-C.sub.4 alkyl,
--Y.sup.2(C(R.sup.6).sub.2).sub.rC(O)NR.sup.gR.sup.g',
--Y.sup.2(C(R.sup.6).sub.2).sub.rS(O).sub.2NR.sup.gR.sup.g',
--(C(R.sup.6).sub.2).sub.rC(O)NR.sup.gR.sup.g',
--(C(R.sup.6).sub.2).sub.rS(O).sub.2NR.sup.gR.sup.g',
--(C(R.sup.6).sub.2).sub.rC(O)NHS(O).sub.2NR.sup.gR.sup.g',
--(C(R.sup.6).sub.2).sub.rCO.sub.2R.sup.i,
--(C(R.sup.6).sub.2).sub.rNH.sub.2CO.sub.2R.sup.x,
--(C(R.sup.6).sub.2).sub.rP(O)(OR.sup.x).sub.2,
--O(C(R.sup.6).sub.2).sub.rP(O)(OR.sup.x).sub.2,
--(C(R.sup.6).sub.2).sub.rS(O).sub.2OH,
--O(C(R.sup.6).sub.2).sub.rS(O).sub.2OH,
--(C(R.sup.6).sub.2).sub.rP(O).sub.2OR.sup.x, or
--O(C(R.sup.6).sub.2).sub.rP(O).sub.2OR.sup.x. In certain
embodiments, B is --(C(R.sup.6).sub.2).sub.rC(O)NR.sup.gR.sup.g'.
In certain embodiments, B is
--(C(R.sup.6).sub.2).sub.rS(O).sub.2NR.sup.gR.sup.g'. In certain
embodiments, B is
--(C(R.sup.6).sub.2).sub.rC(O)NHS(O).sub.2NR.sup.gR.sup.g'.
[0203] In certain embodiments, B is
--(C(R.sup.6).sub.2).sub.rS(O).sub.2OC.sub.1-C.sub.4 alkyl. In
certain embodiments, B is
--O(C(R.sup.6).sub.2).sub.rS(O).sub.2OC.sub.1-C.sub.4 alkyl. In
certain embodiments, B is
--Y.sup.2(C(R.sup.6).sub.2).sub.rC(O)NR.sup.gR.sup.g'. In certain
embodiments, B is
--Y.sup.2(C(R.sup.6).sub.2).sub.rS(O).sub.2NR.sup.gR.sup.g'. In
certain embodiments, B is --(CH.sub.2).sub.rC(O)NR.sup.gR.sup.g'.
In certain embodiments, B is
--(CH.sub.2).sub.rS(O).sub.2NR.sup.gR.sup.g'. In certain
embodiments, B is
--(CH.sub.2).sub.rC(O)NHS(O).sub.2NR.sup.gR.sup.g'. In certain
embodiments, B is --(C(R.sup.6).sub.2).sub.rCO.sub.2R.sup.i. In
certain embodiments, B is
--(C(R.sup.6).sub.2).sub.rNH.sub.2CO.sub.2R.sup.x. In certain
embodiments, B is --(C(R.sup.6).sub.2).sub.rP(O)(OR.sup.x).sub.2.
In certain embodiments, B is
--O(C(R.sup.6).sub.2).sub.rP(O)(OR.sup.x).sub.2. In certain
embodiments, B is --(C(R.sup.6).sub.2).sub.rS(O).sub.2OH. In
certain embodiments, B is --O(C(R.sup.6).sub.2).sub.rS(O).sub.2OH.
In certain embodiments, B is
--(C(R.sup.6).sub.2).sub.rP(O).sub.2OR.sup.x. In certain
embodiments, B is
--O(C(R.sup.6).sub.2).sub.rP(O).sub.2OR.sup.x.
[0204] As described above, C is --(CH.sub.2).sub.rCN,
--(CH.sub.2).sub.sOH, halogen,
--(C(R.sup.6).sub.2).sub.rC.sub.6-C.sub.10 aryl,
--(C(R.sup.6).sub.2).sub.rS--C.sub.6-C.sub.10 aryl,
--(C(R.sup.6).sub.2).sub.rheteroaryl,
--O(C(R.sup.6).sub.2).sub.rheteroaryl,
--O(C(R.sup.6).sub.2).sub.rheterocycloalkyl,
--O(C(R.sup.6).sub.2).sub.rOH, --OR.sup.y,
--(C(R.sup.6).sub.2).sub.rC(O)NHCN, --CH.dbd.CHCO.sub.2R.sup.x, or
--(C(R.sup.6).sub.2).sub.rC(O)NHS(O).sub.2C.sub.1-C.sub.4 alkyl,
wherein the aryl and heteroaryl are substituted with one to three
substituents each independently selected from C.sub.1-C.sub.6
alkyl, C.sub.1-C.sub.6 haloalkyl, halogen, and OH, and wherein the
heterocycloalkyl is substituted with one to two .dbd.O or
.dbd.S;
[0205] In certain embodiments, C is --(CH.sub.2).sub.rCN. In
certain embodiments, C is --(CH.sub.2).sub.sOH. In certain
embodiments, C is halogen. In certain embodiments, C is
--(C(R).sub.2).sub.rC.sub.6-C.sub.10 aryl. In certain embodiments,
C is --(C(R.sup.6).sub.2).sub.rS--C.sub.6-C.sub.10 aryl. In certain
embodiments, C is --(C(R.sup.6).sub.2).sub.rheteroaryl. In certain
embodiments, C is --O(C(R.sup.6).sub.2).sub.rheteroaryl. In certain
embodiments, C is-O(C(R.sup.6).sub.2).sub.rheterocycloalkyl. In
certain embodiments, C is --O(C(R.sup.6).sub.2).sub.rOH. In certain
embodiments, C is --OR.sup.y. In certain embodiments, C is
--(C(R.sup.6).sub.2).sub.rC(O)NHCN. In certain embodiments, C is
--CH.dbd.CHCO.sub.2R.sup.x. In certain embodiments, C is
--(C(R.sup.6).sub.2).sub.rC(O)NHS(O).sub.2C.sub.1-C.sub.4 alkyl. In
the above, the aryl and heteroaryl are substituted with one to
three substituents each independently selected from C.sub.1-C.sub.6
alkyl, C.sub.1-C.sub.6 haloalkyl, halogen, and OH, and wherein the
heterocycloalkyl is substituted with one to two .dbd.O or
.dbd.S.
[0206] As described above, each R.sup.6 is independently at each
occurrence H or C.sub.1-C.sub.4 alkyl. In certain embodiments,
R.sup.6 is H. In certain embodiments, R.sup.6 is C.sub.1-C.sub.4
alkyl.
[0207] As described above, each R.sup.x is independently at each
occurrence H, C.sub.1-C.sub.6 alkyl, or C.sub.6-C.sub.10 aryl. In
certain embodiments, R.sup.x is H. In certain embodiments, R.sup.x
is C.sub.1-C.sub.6 alkyl. In certain embodiments, R.sup.x is
C.sub.6-C.sub.10 aryl.
[0208] As described above, each Y.sup.2 is independently O, NH or
S. In certain embodiments, Y.sup.2 is O. In certain embodiments,
Y.sup.2 is NH. In certain embodiments, Y.sup.2 is S.
[0209] As described above, each r independently is 0, 1 or 2. In
certain embodiments, r is 0. In certain embodiments, r is 1. In
certain embodiments, r is 2.
[0210] As described above, s is 1 or 2. In certain embodiments, s
is 1. In certain embodiments, s is 2.
[0211] As described above, R.sup.g is H, C.sub.1-C.sub.6 alkyl, OH,
--S(O).sub.2(C.sub.1-C.sub.6 alkyl), or
--S(O).sub.2N(C.sub.1-C.sub.6 alkyl).sub.2.
[0212] As described above, R.sup.g is H, C.sub.1-C.sub.6 alkyl,
C.sub.3-C.sub.7 cycloalkyl, 4- to 7-membered heterocycloalkyl ring
comprising 1-3 heteroatoms selected from N, O and S,
C.sub.6-C.sub.10 aryl, or 5- to 7-membered heteroaryl comprising
1-3 heteroatoms selected from N, O and S, wherein the alkyl is
optionally substituted with one or more substituents independently
selected from halogen and --OH, and wherein the cycloalkyl,
heterocycloalkyl, aryl and heteroaryl are optionally substituted
with one or more substituents independently selected from
C.sub.1-C.sub.6 alkyl, halogen, and --OH.
[0213] In some embodiments, the present disclosure provides a
compound of formula (I) having one, two, or three of the following
features:
a) W is N;
b) R.sup.c is CN;
[0214] c) R.sup.d is 5- or 6-membered heteroaryl, such as
thiophenyl; d) L is
--(C(R.sup.5).sub.2).sub.mY.sup.1(C(R.sup.5).sub.2).sub.p, such as
--SCH.sub.2--; e) R.sup.1 is phenylene; f) R.sup.7 is A, such as
COOH or tetrazole.
[0215] In some embodiments, the present disclosure provides a
compound of formula (I) having one, two, or three of the following
features:
a) W is N;
[0216] b) R.sup.d is CF.sub.3; c) L is
--(C(R.sup.5).sub.2).sub.mY.sup.1(C(R.sup.5).sub.2).sub.p, such as
--SCH.sub.2--; d) R.sup.1 is phenylene; e) R.sup.7 is A, such as
COOH or tetrazole.
[0217] In some embodiments, the present disclosure provides a
compound of formula (I) having one, two, or three of the following
features:
a) W is N;
b) R.sup.c is CN;
[0218] c) R.sup.d is 5- or 6-membered heteroaryl, such as
thiophenyl; d) L is
--(C(R.sup.5).sub.2).sub.mY.sup.1(C(R.sup.5).sub.2).sub.p, such as
--SCH.sub.2--; e) R.sup.1 is absent; f) R.sup.7 is A, such as COOH
or tetrazole.
[0219] In some embodiments, the present disclosure provides a
compound of formula (I) having one, two, or three of the following
features:
a) W is N;
[0220] b) R.sup.d is CF.sub.3; c) L is
--(C(R.sup.5).sub.2).sub.mY.sup.1(C(R.sup.5).sub.2).sub.p, such as
--SCH.sub.2--; d) R.sup.1 is absent; e) R.sup.7 is A, such as COOH
or tetrazole.
[0221] In some embodiments, the present disclosure provides a
compound of formula (I) having one, two, or three of the following
features:
a) W is C;
[0222] b) R.sup.d is --(C(R.sup.6).sub.2).sub.tC.sub.6-C.sub.10
aryl or --(C(R.sup.6).sub.2).sub.t-5- or 6-membered heteroaryl); c)
L is --(C(R.sup.5).sub.2).sub.mY.sup.1(C(R.sup.5).sub.2).sub.p,
such as --SCH.sub.2--; d) R.sup.1 is phenylene; e) R.sup.7 is A,
such as COOH or tetrazole.
[0223] In some embodiments, the present disclosure provides a
compound of formula (I) having one, two, or three of the following
features:
a) W is C;
[0224] b) R.sup.d is --CF.sub.3; c) L is
--(C(R.sup.5).sub.2).sub.mY.sup.1(C(R.sup.5).sub.2).sub.p, such as
--SCH.sub.2--; d) R.sup.1 is phenylene; e) R.sup.7 is A, such as
COOH or tetrazole.
[0225] In some embodiments, the present disclosure provides a
compound of formula (I) having one, two, or three of the following
features:
a) W is N;
b) R.sup.c is CN;
[0226] c) R.sup.d is 5- or 6-membered heteroaryl, such as
thiophenyl; d) L is
--(C(R.sup.5).sub.2).sub.mY.sup.1(C(R.sup.5).sub.2).sub.p, such as
--SCH.sub.2--; e) R.sup.1 is phenylene; f) R.sup.7 is A, such as
--(C(R.sup.6).sub.2).sub.rCO.sub.2R.sup.x or
--(CH.sub.2).sub.rtetrazole.
[0227] In some embodiments, the present disclosure provides a
compound of formula (I) having one, two, or three of the following
features:
a) W is N;
[0228] b) R.sup.d is CF.sub.3; c) L is
--(C(R.sup.5).sub.2).sub.mY.sup.1(C(R.sup.5).sub.2).sub.p, such as
--SCH.sub.2--; d) R.sup.1 is phenylene; e) R.sup.7 is A, such as
--(C(R.sup.6).sub.2).sub.rCO.sub.2R.sup.x or
--(CH.sub.2).sub.rtetrazole.
[0229] In some embodiments, the present disclosure provides a
compound of formula (I) having one, two, or three of the following
features:
a) W is N;
b) R.sup.c is CN;
[0230] c) R.sup.d is 5- or 6-membered heteroaryl, such as
thiophenyl; d) L is
--(C(R.sup.5).sub.2).sub.mY.sup.1(C(R.sup.5).sub.2).sub.p, such as
--SCH.sub.2--; e) R.sup.1 is absent; f) R.sup.7 is A, such as
--(C(R.sup.6).sub.2).sub.rCO.sub.2R.sup.x or
--(CH.sub.2).sub.rtetrazole.
[0231] In some embodiments, the present disclosure provides a
compound of formula (I) having one, two, or three of the following
features:
a) W is N;
[0232] b) R.sup.d is CF.sub.3; c) L is
--(C(R.sup.5).sub.2).sub.mY.sup.1(C(R.sup.5).sub.2).sub.p, such as
--SCH.sub.2--; d) R.sup.1 is absent; e) R.sup.7 is A, such as
--(C(R.sup.6).sub.2).sub.rCO.sub.2R.sup.x or
--(CH.sub.2).sub.rtetrazole.
[0233] In some embodiments, the present disclosure provides a
compound of formula (I) having one, two, or three of the following
features:
a) W is C;
[0234] b) R.sup.d is --(C(R.sup.6).sub.2).sub.tC.sub.6-C.sub.10
aryl or --(C(R.sup.6).sub.2).sub.t-5- or 6-membered heteroaryl); c)
L is --(C(R.sup.5).sub.2).sub.mY.sup.1(C(R.sup.5).sub.2).sub.p,
such as --SCH.sub.2--; d) R.sup.1 is phenylene; e) R.sup.7 is A,
such as --(C(R.sup.6).sub.2).sub.rCO.sub.2R.sup.x or
--(CH.sub.2).sub.rtetrazole.
[0235] In some embodiments, the present disclosure provides a
compound of formula (I) having one, two, or three of the following
features:
a) W is C;
[0236] b) R.sup.d is --CF.sub.3; c) L is
--(C(R.sup.5).sub.2).sub.mY.sup.1(C(R.sup.5).sub.2).sub.p, such as
--SCH.sub.2--; d) R.sup.1 is phenylene; e) R.sup.7 is A, such as
--(C(R.sup.6).sub.2).sub.rCO.sub.2R.sup.x or
--(CH.sub.2).sub.rtetrazole.
[0237] In certain embodiments, with certain above features for
Formula (I), the present disclosure provides a compound of formula
(Ia) having at least one of the following features:
##STR00019##
and pharmaceutically salts and tautomers thereof, wherein a)
R.sup.d is 5- or 6-membered heteroaryl; b) L is
--(C(R.sup.5).sub.2).sub.mY.sup.1(C(R.sup.5).sub.2).sub.p;
c) R.sup.7 is A or C;
[0238] d) X, R.sup.d, R.sup.f, R.sup.j, A, R.sup.5, Y.sup.1, m, and
p are defined for Formula (I). In certain embodiments, R.sup.d is
thiophenyl. In certain embodiments, L is --SCH.sub.2-- or
--NHCH.sub.2--. In certain embodiments, R.sup.7 is C. In certain
embodiments, C is --(C(R.sup.6).sub.2).sub.rC.sub.6-C.sub.10 aryl,
substituted with one to three substituents each independently
selected from C.sub.1-C.sub.6 alkyl, C.sub.1-C.sub.6 haloalkyl,
halogen and OH. In certain embodiments, R.sup.7 is A. In certain
embodiments, A is --(C(R.sup.6).sub.2).sub.rCO.sub.2R.sup.x or
--(CH.sub.2).sub.rtetrazole, wherein --(CH.sub.2).sub.rtetrazole is
optionally substituted with C.sub.1-C.sub.6 alkyl. In certain
embodiments, A is --(C(R.sup.6).sub.2).sub.rCOOH or
--(CH.sub.2).sub.rtetrazole, wherein --(CH.sub.2).sub.rtetrazole is
optionally substituted with C.sub.1-C.sub.6 alkyl. In certain
embodiments, A is --COOH, --CH.sub.2COOH, -tetrazole, or
--(CH.sub.2)tetrazole, wherein tetrazole and
--(CH.sub.2).sub.rtetrazole are optionally substituted with
C.sub.1-C.sub.6 alkyl. In certain embodiments, Formula (Ia) has
one, two, three or four of the features (a) to (d).
[0239] In certain embodiments, with certain above features for
Formula (I), the present disclosure provides a compound of formula
(Ib) having at least one of the following features:
##STR00020##
and pharmaceutically salts and tautomers thereof, wherein a) L is
--(C(R.sup.5).sub.2).sub.mY.sup.1(C(R.sup.5).sub.2).sub.p;
b) R.sup.7 is A;
[0240] c) X, R.sup.c, R.sup.f, R.sup.j, A, R.sup.5, Y.sup.1, m, and
p are defined for Formula (I).
[0241] In certain embodiments, L is --SCH.sub.2-- or
--NHCH.sub.2--. In certain embodiments, A is
--(C(R.sup.6).sub.2).sub.rCO.sub.2R.sup.x or
--(CH.sub.2).sub.rtetrazole, wherein --(CH.sub.2).sub.rtetrazole is
optionally substituted with C.sub.1-C.sub.6 alkyl. In certain
embodiments, A is --(C(R.sup.6).sub.2).sub.rCOOH or
--(CH.sub.2).sub.rtetrazole, wherein --(CH.sub.2).sub.rtetrazole is
optionally substituted with C.sub.1-C.sub.6 alkyl. In certain
embodiments, A is --COOH, --CH.sub.2COOH, -tetrazole, or
--(CH.sub.2)tetrazole, wherein tetrazole and
--(CH.sub.2).sub.rtetrazole are optionally substituted with
C.sub.1-C.sub.6 alkyl. In certain embodiments, Formula (Ib) has
one, two, or three of the features (a) to (c).
[0242] In certain embodiments, with certain above features for
Formula (I), the present disclosure provides a compound of formula
(Ic) having at least one of the following features:
##STR00021##
and pharmaceutically salts and tautomers thereof, wherein a)
R.sup.d is 5- or 6-membered heteroaryl; b) L is
--(C(R.sup.5).sub.2).sub.mY.sup.1(C(R.sup.5).sub.2).sub.p;
c) R.sup.7 is A;
[0243] d) X, R.sup.d, R.sup.f, R.sup.j, A, R.sup.5, Y.sup.1, m, and
p are defined for Formula (I). In certain embodiments, R.sup.d is
thiophenyl. In certain embodiments, L is --SCH.sub.2-- or
--NHCH.sub.2--. In certain embodiments, A is
--(C(R.sup.6).sub.2).sub.rCO.sub.2R.sup.x or
--(CH.sub.2).sub.rtetrazole, wherein --(CH.sub.2).sub.rtetrazole is
optionally substituted with C.sub.1-C.sub.6 alkyl. In certain
embodiments, A is --(C(R.sup.6).sub.2).sub.rCOOH or
--(CH.sub.2).sub.rtetrazole, wherein --(CH.sub.2).sub.rtetrazole is
optionally substituted with C.sub.1-C.sub.6 alkyl. In certain
embodiments, A is --COOH, --CH.sub.2COOH, -tetrazole, or
--(CH.sub.2)tetrazole, wherein tetrazole and
--(CH.sub.2).sub.rtetrazole are optionally substituted with
C.sub.1-C.sub.6 alkyl. In certain embodiments, Formula (Ic) has
one, two, three or four of the features (a) to (d).
[0244] In certain embodiments, with certain above features for
Formula (I), the present disclosure provides a compound of formula
(Id) having at least one of the following features:
##STR00022##
and pharmaceutically salts and tautomers thereof, wherein a) L is
--(C(R.sup.5).sub.2).sub.mY.sup.1(C(R.sup.5).sub.2).sub.p;
b) R.sup.7 is A;
[0245] c) X, R.sup.d, R.sup.f, R.sup.j, A, R.sup.5, Y.sup.1, m, and
p are defined for Formula (I). In certain embodiments, L is
--SCH.sub.2-- or --NHCH.sub.2--. In certain embodiments, A is
--(C(R.sup.6).sub.2).sub.rCO.sub.2R.sup.x or
--(CH.sub.2).sub.rtetrazole, wherein --(CH.sub.2).sub.rtetrazole is
optionally substituted with C.sub.1-C.sub.6 alkyl. In certain
embodiments, A is --(C(R.sup.6).sub.2).sub.rCOOH or
--(CH.sub.2).sub.rtetrazole, wherein --(CH.sub.2).sub.rtetrazole is
optionally substituted with C.sub.1-C.sub.6 alkyl. In certain
embodiments, A is --COOH, --CH.sub.2COOH, -tetrazole, or
--(CH.sub.2)tetrazole, wherein tetrazole and
--(CH.sub.2).sub.rtetrazole are optionally substituted with
C.sub.1-C.sub.6 alkyl. In certain embodiments, Formula (Id) has
one, two, or three of the features (a) to (c).
[0246] In certain embodiments, with certain above features for
Formula (I), the present disclosure provides a compound of formula
(Ie) having at least one of the following features:
##STR00023##
and pharmaceutically salts and tautomers thereof, wherein a)
R.sup.d is --(C(R.sup.6).sub.2).sub.tC.sub.6-C.sub.10 aryl or
--(C(R.sup.6).sub.2).sub.t-5- or 6-membered heteroaryl); b) L is
--(C(R.sup.5).sub.2).sub.mY.sup.1(C(R.sup.5).sub.2).sub.p;
c) R.sup.7 is A.
[0247] d) X, R.sup.d, R.sup.f, R.sup.j, A, R.sup.5, Y.sup.1, m, and
p are defined for Formula (I).
[0248] In certain embodiments, L is --SCH.sub.2-- or
--NHCH.sub.2--. In certain embodiments, A is
--(C(R.sup.6).sub.2).sub.rCO.sub.2R.sup.x or
--(CH.sub.2).sub.rtetrazole, wherein --(CH.sub.2).sub.rtetrazole is
optionally substituted with C.sub.1-C.sub.6 alkyl. In certain
embodiments, A is --(C(R.sup.6).sub.2).sub.rCOOH or
--(CH.sub.2).sub.rtetrazole, wherein --(CH.sub.2).sub.rtetrazole is
optionally substituted with C.sub.1-C.sub.6 alkyl. In certain
embodiments, A is --COOH, --CH.sub.2COOH, -tetrazole, or
--(CH.sub.2)tetrazole, wherein tetrazole and
--(CH.sub.2).sub.rtetrazole are optionally substituted with
C.sub.1-C.sub.6 alkyl. In certain embodiments, Formula (Ie) has
one, two, three, or four of the features (a) to (d).
[0249] In certain embodiments, with certain above features for
Formula (I), the present disclosure provides a compound of formula
(If) having at least one of the following features:
##STR00024##
and pharmaceutically salts and tautomers thereof, wherein a) L is
--(C(R.sup.5).sub.2).sub.mY.sup.1(C(R.sup.5).sub.2).sub.p;
b) R.sup.7 is A;
[0250] c) X, R.sup.c, R.sup.f, R.sup.i, A, R.sup.5, Y.sup.1, m, and
p are defined for Formula (I). In certain embodiments, L is
--SCH.sub.2-- or --NHCH.sub.2--. In certain embodiments, A is
--(C(R.sup.6).sub.2).sub.rCO.sub.2R.sup.x or
--(CH.sub.2).sub.rtetrazole, wherein --(CH.sub.2).sub.rtetrazole is
optionally substituted with C.sub.1-C.sub.6 alkyl. In certain
embodiments, A is --(C(R.sup.6).sub.2).sub.rCOOH or
--(CH.sub.2).sub.rtetrazole, wherein --(CH.sub.2).sub.rtetrazole is
optionally substituted with C.sub.1-C.sub.6 alkyl. In certain
embodiments, A is --COOH, --CH.sub.2COOH, -tetrazole, or
--(CH.sub.2)tetrazole, wherein tetrazole and
--(CH.sub.2).sub.rtetrazole are optionally substituted with
C.sub.1-C.sub.6 alkyl. In certain embodiments, R.sup.c is CN. In
certain embodiments, Formula (If) has one, two, or three of the
features (a) to (c).
[0251] In certain embodiments, with certain above features for
Formula (I), the present disclosure provides a compound of formula
(Ig) having at least one of the following features:
##STR00025##
and pharmaceutically salts and tautomers thereof, wherein a) L is
--(C(R.sup.5).sub.2).sub.mY.sup.1(C(R.sup.5).sub.2).sub.p;
b) R.sup.7 is A;
[0252] c) X, R.sup.d, R.sup.f, R.sup.j, A, R.sup.5, Y.sup.1, m, and
p are defined for Formula (I). In certain embodiments, L is
--SCH.sub.2-- or --NHCH.sub.2--. In certain embodiments, A is
--(C(R.sup.6).sub.2).sub.rCO.sub.2R.sup.x or
--(CH.sub.2).sub.rtetrazole, wherein --(CH.sub.2).sub.rtetrazole is
optionally substituted with C.sub.1-C.sub.6 alkyl. In certain
embodiments, A is --(C(R.sup.6).sub.2).sub.rCOOH or
--(CH.sub.2).sub.rtetrazole, wherein --(CH.sub.2).sub.rtetrazole is
optionally substituted with C.sub.1-C.sub.6 alkyl. In certain
embodiments, A is --COOH, --CH.sub.2COOH, -tetrazole, or
--(CH.sub.2)tetrazole, wherein tetrazole and
--(CH.sub.2).sub.rtetrazole are optionally substituted with
C.sub.1-C.sub.6 alkyl. In certain embodiments, Formula (Ig) has
one, two, or three of the features (a) to (c).
[0253] In some embodiments, the compound of Formula (I) is a
compound selected from:
TABLE-US-00001 Cpd No. Structure I-1 ##STR00026## I-2 ##STR00027##
I-3 ##STR00028## I-4 ##STR00029## I-5 ##STR00030## I-6 ##STR00031##
I-7 ##STR00032## I-8 ##STR00033## I-9 ##STR00034## I-10
##STR00035## I-11 ##STR00036## I-12 ##STR00037## I-13 ##STR00038##
I-14 ##STR00039## I-15 ##STR00040## I-16 ##STR00041## I-17
##STR00042## I-18 ##STR00043## I-19 ##STR00044## I-20 ##STR00045##
I-21 ##STR00046## I-22 ##STR00047## I-23 ##STR00048## I-24
##STR00049## I-25 ##STR00050## I-26 ##STR00051## I-27 ##STR00052##
I-28 ##STR00053## I-29 ##STR00054## I-30 ##STR00055## I-31
##STR00056## I-32 ##STR00057## I-33 ##STR00058## I-34 ##STR00059##
I-35 ##STR00060## I-36 ##STR00061##
or a pharmaceutically acceptable salt or tautomer thereof.
[0254] In some embodiments, the compound of Formula (I) or (II) is
a compound selected from:
TABLE-US-00002 I-37 ##STR00062## I-38 ##STR00063## I-39
##STR00064## I-40 ##STR00065## I-41 ##STR00066## I-42 ##STR00067##
I-43 ##STR00068## I-44 ##STR00069## I-45 ##STR00070## I-46
##STR00071## I-47 ##STR00072## I-48 ##STR00073## I-49 ##STR00074##
I-50 ##STR00075## I-51 ##STR00076##
[0255] In some embodiments, the compound of Formula (I) is a
compound, or a pharmaceutically acceptable salt or tautomer
thereof, selected from:
TABLE-US-00003 ##STR00077## ##STR00078## ##STR00079## ##STR00080##
##STR00081## ##STR00082## ##STR00083## ##STR00084## ##STR00085##
##STR00086## ##STR00087## ##STR00088## ##STR00089## ##STR00090##
##STR00091## ##STR00092## ##STR00093## ##STR00094## ##STR00095##
##STR00096## ##STR00097## ##STR00098## ##STR00099## ##STR00100##
##STR00101## ##STR00102## ##STR00103## ##STR00104## ##STR00105##
##STR00106## ##STR00107## ##STR00108## ##STR00109## ##STR00110##
##STR00111## ##STR00112## ##STR00113## ##STR00114## ##STR00115##
##STR00116## ##STR00117## ##STR00118## ##STR00119## ##STR00120##
##STR00121## ##STR00122## ##STR00123## ##STR00124## ##STR00125##
##STR00126## ##STR00127## ##STR00128## ##STR00129## ##STR00130##
##STR00131## ##STR00132## ##STR00133## ##STR00134## ##STR00135##
##STR00136## ##STR00137## ##STR00138## ##STR00139## ##STR00140##
##STR00141## ##STR00142## ##STR00143## ##STR00144## ##STR00145##
##STR00146## ##STR00147## ##STR00148## ##STR00149## ##STR00150##
##STR00151## ##STR00152##
or a pharmaceutically acceptable salt or tautomer thereof.
[0256] In some embodiments, the compound of Formula (I) or (II) is
a compound, or a pharmaceutically acceptable salt or tautomer
thereof, selected from:
##STR00153##
[0257] It should be understood, that such references are intended
to encompass not only the above general formula, but also each and
every of the embodiments, etc. discussed in the following. It
should also be understood, that unless stated to the opposite, such
references also encompass isomers, mixtures of isomers,
pharmaceutically acceptable salts, solvates and prodrugs of the
compounds of Formula (I) or (II).
Methods for the Preparation of Compounds
[0258] The compounds of the present disclosure (e.g., compounds of
Formula (I)) can be prepared in a number of ways well known to
those skilled in the art of organic synthesis. By way of example,
compounds of the present disclosure can be synthesized using the
methods described below, together with synthetic methods known in
the art of synthetic organic chemistry, or variations thereon as
appreciated by those skilled in the art. Preferred methods include
but are not limited to those methods described below. The final
products of the reactions described herein may be isolated by
conventional techniques, e.g., by extraction, crystallisation,
distillation, chromatography, etc.
[0259] Compounds of the present disclosure can be synthesized by
following the steps outlined in General Scheme A to F which
comprise different sequences of assembling intermediates Ia-Ih and
Ij-Io. Starting materials are either commercially available or made
by known procedures in the reported literature or as illustrated.
Useful steps that may be used in the preparation steps of the
compounds will be known to the skilled person. The method below is
given as a non-limiting example on how the compounds may be
prepared.
##STR00154##
[0260] wherein R.sup.1, R.sup.c, R.sup.d, and L are defined as in
Formula (I).
[0261] The general way of preparing compounds of Formula (I) by
using intermediates Ia, and Ib is outlined in General Scheme A.
Coupling of Ta with Ib using a base, i.e., potassium carbonate
(K.sub.2CO.sub.3), in a solvent, i.e., acetonitrile (CH.sub.3CN),
optionally at elevated temperature provides the desired produce of
Formula (I). Bases that can be used include, but are not limited
to, sodium carbonate (Na.sub.2CO.sub.3), potassium carbonate
(K.sub.2CO.sub.3), N,N-diisopropylethylamine (DIPEA) and
triethylamine. Solvents used in the coupling reaction can be polar
or non-polar solvents. For example, the solvent can be acetonitrile
(CH.sub.3CN), acetone, or dimethylsulfoxide (DMSO).
##STR00155##
[0262] wherein X is a good leaving group, i.e., Cl, Br,
--SCH.sub.3, or S(O).sub.2CH.sub.3, and R.sup.1, R.sup.2, R.sup.c,
R.sup.d, and p are defined as in Formula (I).
[0263] Alternatively, compounds of Formula (I) can be prepared
using intermediates Ic and Id as outlined in General Scheme B.
Amination of Intermediate Ic with Ie using a base, i.e., sodium
hydroxide (NaOH), potassium hydroxide (KOH), etc., in a solvent,
i.e., methanol (MeOH), ethanol (EtOH), water (H.sub.2O), etc.,
provides compounds of Formula (I).
##STR00156##
[0264] wherein X is a good leaving group, i.e., Cl, Br,
--SCH.sub.3, or S(O).sub.2CH.sub.3, and R.sup.1, R.sup.2, R.sup.c,
R.sup.d, and p are defined as in Formula (I).
[0265] Compounds of Formula (I) can also be prepared using
intermediates Ie and If as outlined in General Scheme C. Amination
of Intermediate Ie with If using a base, i.e., sodium hydroxide
(NaOH), potassium hydroxide (KOH), etc., in a solvent, i.e.,
methanol (MeOH), ethanol (EtOH), water (H.sub.2O), etc., provides
compounds of Formula (I).
##STR00157##
[0266] wherein and R.sup.1, R.sup.c, and R.sup.d are defined as in
Formula (I).
[0267] Alternatively, compounds of Formula (I) can also be prepared
using intermediates Ig, Ih, Ij, Ik, and Im as outlined in General
Scheme D. Olefination of intermediate Ig using a base i.e.,
potassium carbonate (K.sub.2CO.sub.3) and diethyl
(cyanomethyl)phosphonate in a solvent, i.e., tetrahydrofuran (THF),
water (H.sub.2O), optionally at an elevated temperature provides
Intermediate Ih. Hydrogenation of Ih using a metal catalyst, i.e.,
palladium on carbon (Pd/C), platinum dioxide (PtO.sub.2), etc, and
hydrogen (H.sub.2) gas in a solvent, i.e., ethanol (EtOH) and/or
tetrahydrofuran (THF), provides Intermediate Ij. Intermediate Ik is
obtained by treating Intermediate Ij with an acid, i.e.,
hydrochloric acid (HCl) in a solvent, i.e., ethanol (EtOH),
dichloromethane (CH.sub.2Cl.sub.2), etc., and then subsequent
treatment with a base, i.e., ammonia (NH.sub.3). Cyclization of
Intermediate Ik and Im using a base, i.e., sodium hydroxide (NaOH),
potassium hydroxide (KOH), etc., in a solvent, i.e.,
dimethylacetamide (DMA), optionally at elevated temperature
provides compounds of Formula (I).
##STR00158##
[0268] wherein and R.sup.1, R.sup.c, and R.sup.d are defined as in
Formula (I).
[0269] Alternatively, compounds of Formula (I) can be prepared
using intermediates In and Io as outlined in General Scheme D.
Acylation of Intermediate In with Io using a base, i.e., sodium
hydroxide (NaOH), potassium hydroxide (KOH), etc., in a solvent,
i.e., methanol (MeOH), ethanol (EtOH), water (H.sub.2O), etc.,
provides compounds of Formula (I).
##STR00159##
[0270] wherein and L, R.sup.c, R.sup.d, R.sup.1, and R.sup.7 are
defined as in Formula (I).
[0271] The general procedure for the synthesis of compounds (e.g.,
I-17 to I-30) with general Formula I include the final coupling
between one equivalent of the corresponding substituted
6-mercapto-2-oxo-4,5-disubstituted-1,2-dihydro-pyridine derivative
and a stoichiometric amount of the L-R.sup.1-R.sup.7 intermediates
using two equivalent of DIPEA as base and acetone as solvent to
provide the final compound.
[0272] Alternatively, certain compounds of Formula (I) or (II) can
be prepared using the schemes shown below and compounds of Formula
(I) or (II) in general can be prepared based on the schemes shown
below.
##STR00160##
##STR00161##
##STR00162##
##STR00163##
##STR00164##
[0273] A mixture of enantiomers, diastereomers, cis/trans isomers
resulting from the process described above can be separated into
their single components by chiral salt technique, chromatography
using normal phase, reverse phase or chiral column, depending on
the nature of the separation.
[0274] It should be understood that in the description and formula
shown above, the various groups R.sup.i, R.sup.2, X, L, Y, R.sup.a,
R.sup.b, R.sup.c, R.sup.d, R.sup.e, R.sup.f, R.sup.x, R.sup.y,
R.sup.z, m, n, p, q, r and other variables are as defined herein
above, except where otherwise indicated. Furthermore, for synthetic
purposes, the compounds of General Schemes A-E are merely
representative with elected radicals to illustrate the general
synthetic methodology of the compounds of Formula (I) as defined
herein.
Pharmaceutical Compositions
[0275] The compound of Formula (I) or (II) may be provided in any
form suitable for the intended administration, in particular
including pharmaceutically acceptable salts, solvates and prodrugs
of the compound of Formula (I) or (II).
[0276] Pharmaceutically acceptable salts refer to salts of the
compounds of Formula (I) or (II) which are considered to be
acceptable for clinical and/or veterinary use. Typical
pharmaceutically acceptable salts include those salts prepared by
reaction of the compounds of Formula (I) or (II) and a mineral or
organic acid or an organic or inorganic base. Such salts are known
as acid addition salts and base addition salts, respectively. It
will be recognized that the particular counter-ion forming a part
of any salt is not of a critical nature, so long as the salt as a
whole is pharmaceutically acceptable and as long as the counter-ion
does not contribute undesired qualities to the salt as a whole.
These salts may be prepared by methods known to the skilled person.
Pharmaceutically acceptable salts are, e.g., those described and
discussed in Remington's Pharmaceutical Sciences, 17. Ed. Alfonso
R. Gennaro (Ed.), Mack Publishing Company, Easton, Pa., U.S.A.,
1985 and more recent editions and in Encyclopedia of Pharmaceutical
Technology.
[0277] Examples of pharmaceutically acceptable addition salts
include acid addition salts formed with inorganic acids, e.g.,
hydrochloric, hydrobromic, sulfuric, nitric, hydroiodic,
metaphosphoric, or phosphoric acid; and organic acids e.g.,
succinic, maleic, acetic, fumaric, citric, tartaric, benzoic,
trifluoroacetic, malic, lactic, formic, propionic, glycolic,
gluconic, camphorsulfuric, isothionic, mucic, gentisic,
isonicotinic, saccharic, glucuronic, furoic, glutamic, ascorbic,
anthranilic, salicylic, phenylacetic, mandelic, embonic (pamoic),
ethanesulfonic, pantothenic, stearic, sulfinilic, alginic and
galacturonic acid; and arylsulfonic, for example benzenesulfonic,
p-toluenesulfonic, methanesulfonic or naphthalenesulfonic acid; and
base addition salts formed with alkali metals and alkaline earth
metals and organic bases such as N,N-dibenzylethylenediamine,
chloroprocaine, choline, diethanolamine, ethylenediamine, meglumine
(N-methylglucamine), lysine and procaine; and internally formed
salts. It should be understood that all references to
pharmaceutically acceptable salts include solvent addition forms
(solvates) or crystal forms (polymorphs) as defined herein, of the
same salt.
[0278] The compound of Formula (I) or (II), or a pharmaceutically
acceptable salt thereof, may be provided in dissoluble or
indissoluble forms together with a pharmaceutically acceptable
solvent such as water, ethanol, and the like. Dissoluble forms may
also include hydrated forms such as the mono-hydrate, the
dihydrate, the hemihydrate, the trihydrate, the tetrahydrate, and
the like.
[0279] The compound of Formula (I) or (II), or a pharmaceutically
acceptable salt thereof, may be provided as a prodrug. The term
"prodrug" used herein is intended to mean a compound which--upon
exposure to certain physiological conditions--will liberate the
compound of Formula (I) or (II), or a pharmaceutically acceptable
salt thereof, which then will be able to exhibit the desired
biological action. A typical example is a labile carbamate of an
amine.
[0280] Since prodrugs are known to enhance numerous desirable
qualities of pharmaceuticals (e.g., solubility, bioavailability,
manufacturing, etc.), the compounds of the present disclosure can
be delivered in prodrug form. Thus, the present disclosure is
intended to cover prodrugs of the presently claimed compounds,
methods of delivering the same and compositions containing the
same. "Prodrugs" are intended to include any covalently bonded
carriers that release an active parent drug of the present
disclosure in vivo when such prodrug is administered to a subject.
Prodrugs in the present disclosure are prepared by modifying
functional groups present in the compound in such a way that the
modifications are cleaved, either in routine manipulation or in
vivo, to the parent compound. Prodrugs include compounds of the
present disclosure wherein a hydroxy, amino, sulfhydryl, carboxy or
carbonyl group is bonded to any group that may be cleaved in vivo
to form a free hydroxyl, free amino, free sulfhydryl, free carboxy
or free carbonyl group, respectively.
[0281] Examples of prodrugs include, but are not limited to, esters
(e.g., acetate, dialkylaminoacetates, formates, phosphates,
sulfates and benzoate derivatives) and carbamates (e.g.,
N,N-dimethylaminocarbonyl) of hydroxy functional groups, esters
(e.g., C.sub.1-6 alkyl esters, e.g., methyl esters, ethyl esters,
2-propyl esters, phenyl esters, 2-aminoethyl esters,
morpholinoethanol esters, etc.) of carboxyl functional groups,
N-acyl derivatives (e.g., N-acetyl) N-Mannich bases, Schiff bases
and enaminones of amino functional groups, oximes, acetals, ketals
and enol esters of ketone and aldehyde functional groups in
compounds of the disclosure, and the like. See Bundegaard, H.,
Design of Prodrugs, p 1-92, Elesevier, New York-Oxford (1985).
[0282] The compounds, or pharmaceutically acceptable salts, esters
or prodrugs thereof, are administered orally, nasally,
transdermally, pulmonary, inhalationally, buccally, sublingually,
intraperintoneally, subcutaneously, intramuscularly, intravenously,
rectally, intrapleurally, intrathecally and parenterally. In one
embodiment, the compound is administered orally. One skilled in the
art will recognize the advantages of certain routes of
administration.
[0283] The dosage regimen utilizing the compounds is selected in
accordance with a variety of factors including type, species, age,
weight, sex and medical condition of the patient; the severity of
the condition to be treated; the route of administration; the renal
and hepatic function of the patient; and the particular compound or
salt thereof employed. An ordinarily skilled physician or
veterinarian can readily determine and prescribe the effective
amount of the drug required to prevent, counter or arrest the
progress of the condition.
[0284] Techniques for formulation and administration of the
disclosed compounds of the disclosure can be found in Remington:
the Science and Practice of Pharmacy, 19.sup.th edition, Mack
Publishing Co., Easton, Pa. (1995). In an embodiment, the compounds
described herein, and the pharmaceutically acceptable salts
thereof, are used in pharmaceutical preparations in combination
with a pharmaceutically acceptable carrier or diluent. Suitable
pharmaceutically acceptable carriers include inert solid fillers or
diluents and sterile aqueous or organic solutions. The compounds
will be present in such pharmaceutical compositions in amounts
sufficient to provide the desired dosage amount in the range
described herein.
[0285] In one aspect of this disclosure, there is provided a
pharmaceutical composition comprising at, as an active ingredient,
at least one compound of Formula (I) or (II), or a pharmaceutically
acceptable salt thereof, as defined herein, and optionally one or
more pharmaceutically acceptable excipients, diluents and/or
carriers. The compounds of Formula (I) or (II), or a
pharmaceutically acceptable salt thereof, may be administered alone
or in combination with pharmaceutically acceptable carriers,
diluents or excipients, in either single or multiple doses.
Suitable pharmaceutically acceptable carriers, diluents and
excipients include inert solid diluents or fillers, sterile aqueous
solutions and various organic solvents.
[0286] A "pharmaceutical composition" is a formulation containing
the compounds of the present disclosure in a form suitable for
administration to a subject. The pharmaceutical compositions may be
formulated with pharmaceutically acceptable carriers or diluents as
well as any other known adjuvants and excipients in accordance with
conventional techniques such as those disclosed in Remington: The
Science and Practice of Pharmacy, 21st Edition, 2000, Lippincott
Williams & Wilkins.
[0287] As used herein, the phrase "pharmaceutically acceptable"
refers to those compounds, materials, compositions, carriers,
and/or dosage forms which are, within the scope of sound medical
judgment, suitable for use in contact with the tissues of human
beings and animals without excessive toxicity, irritation, allergic
response, or other problem or complication, commensurate with a
reasonable benefit/risk ratio.
[0288] "Pharmaceutically acceptable excipient" means an excipient
that is useful in preparing a pharmaceutical composition that is
generally safe, non-toxic and neither biologically nor otherwise
undesirable, and includes excipient that is acceptable for
veterinary use as well as human pharmaceutical use. A
"pharmaceutically acceptable excipient" as used in the
specification and claims includes both one and more than one such
excipient.
[0289] The pharmaceutical compositions formed by combining a
compound of Formula (I) or (II), or a pharmaceutically acceptable
salt thereof, as defined herein, with pharmaceutically acceptable
carriers, diluents or excipients can be readily administered in a
variety of dosage forms such as tablets, powders, lozenges, syrups,
suppositories, injectable solutions and the like. In powders, the
carrier is a finely divided solid such as talc or starch which is
in a mixture with the finely divided active component. In tablets,
the active component is mixed with the carrier having the necessary
binding properties in suitable proportions and compacted in the
shape and size desired.
[0290] The pharmaceutical compositions may be specifically prepared
for administration by any suitable route such as the oral and
parenteral (including subcutaneous, intramuscular, intrathecal,
intravenous and intradermal) route. It will be appreciated that the
preferred route will depend on the general condition and age of the
subject to be treated, the nature of the condition to be treated
and the active ingredient chosen.
[0291] Pharmaceutical compositions for oral administration include
solid dosage forms such as capsules, tablets, dragees, pills,
lozenges, powders, and granules. Where appropriate, they can be
prepared with coatings such as enteric coatings or they can be
prepared so as to provide controlled release of the active
ingredient such as sustained or prolonged release according to
methods well known in the art.
[0292] For oral administration in the form of a tablet or capsule,
a compound of Formula (I) or (II), or a pharmaceutically acceptable
salt thereof, as defined herein, may suitably be combined with an
oral, non-toxic, pharmaceutically acceptable carrier such as
ethanol, glycerol, water, or the like. Furthermore, suitable
binders, lubricants, disintegrating agents, flavouring agents, and
colourants may be added to the mixture, as appropriate. Suitable
binders include, e.g., lactose, glucose, starch, gelatin, acacia
gum, tragacanth gum, sodium alginate, carboxymethylcellulose,
polyethylene glycol, waxes, or the like. Lubricants include, e.g.,
sodium oleate, sodium stearate, magnesium stearate, sodium
benzoate, sodium acetate, sodium chloride, or the like.
Disintegrating agents include, e.g., starch, methyl cellulose,
agar, bentonite, xanthan gum, sodium starch glycolate,
crospovidone, croscarmellose sodium, or the like. Additional
excipients for capsules include macrogels or lipids.
[0293] For the preparation of solid compositions such as tablets,
the active compound of Formula (I) or (II), or a pharmaceutically
acceptable salt thereof, is mixed with one or more excipients, such
as the ones described above, and other pharmaceutical diluents such
as water to make a solid pre-formulation composition containing a
homogenous mixture of a compound of Formula (I) or (II), or a
pharmaceutically acceptable salt thereof. The term "homogenous" is
understood to mean that the compound of Formula (I) or (II), or a
pharmaceutically acceptable salt thereof, is dispersed evenly
throughout the composition so that the composition may readily be
subdivided into equally effective unit dosage forms such as tablets
or capsules.
[0294] Liquid compositions for either oral or parenteral
administration of the compound of Formula (I) or (II), or a
pharmaceutically acceptable salt thereof, include, e.g., aqueous
solutions, syrups, elixirs, aqueous or oil suspensions and emulsion
with edible oils such as cottonseed oil, sesame oil, coconut oil,
or peanut oil. Suitable dispersing or suspending agents for aqueous
suspensions include synthetic or natural gums such as tragacanth,
alginate, acacia, dextran, sodium carboxymethylcellulose, gelatin,
methylcellulose, or polyvinylpyrrolidone.
[0295] Pharmaceutical compositions for parenteral administration
include sterile aqueous and non-aqueous injectable solutions,
dispersions, suspensions or emulsions as well as sterile powders to
be reconstituted in sterile injectable solutions or dispersions
prior to use.
[0296] For intravenous administration, suitable carriers include
physiological saline, bacteriostatic water, Cremophor EL.TM. (BASF,
Parsippany, N.J.) or phosphate buffered saline (PBS). In all cases,
the composition must be sterile and should be fluid to the extent
that easy syringeability exists. It must be stable under the
conditions of manufacture and storage and must be preserved against
the contaminating action of microorganisms such as bacteria and
fungi. The carrier can be a solvent or dispersion medium
containing, for example, water, ethanol, polyol (for example,
glycerol, propylene glycol, and liquid polyethylene glycol, and the
like), and suitable mixtures thereof. The proper fluidity can be
maintained, for example, by the use of a coating such as lecithin,
by the maintenance of the required particle size in the case of
dispersion and by the use of surfactants. Prevention of the action
of microorganisms can be achieved by various antibacterial and
antifungal agents, for example, parabens, chlorobutanol, phenol,
ascorbic acid, thimerosal, and the like. In many cases, it will be
preferable to include isotonic agents, for example, sugars,
polyalcohols such as manitol, sorbitol, and sodium chloride in the
composition. Prolonged absorption of the injectable compositions
can be brought about by including in the composition an agent which
delays absorption, for example, aluminum monostearate and
gelatin.
[0297] The preparation of all these solutions under sterile
conditions is readily accomplished by standard pharmaceutical
techniques well known to those skilled in the art.
[0298] For example, sterile injectable solutions can be prepared by
incorporating the active compound in the required amount in an
appropriate solvent with one or a combination of ingredients
enumerated above, as required, followed by filtered sterilization.
Generally, dispersions are prepared by incorporating the active
compound into a sterile vehicle that contains a basic dispersion
medium and the required other ingredients from those enumerated
above. In the case of sterile powders for the preparation of
sterile injectable solutions, methods of preparation are vacuum
drying and freeze-drying that yields a powder of the active
ingredient plus any additional desired ingredient from a previously
sterile-filtered solution thereof. Depot injectable compositions
are also contemplated as being within the scope of the present
disclosure.
[0299] For parenteral administration, solutions containing a
compound of Formula (I) or (II), or a pharmaceutically acceptable
salt thereof, in sesame or peanut oil, aqueous propylene glycol, or
in sterile aqueous solution may be employed. Such aqueous solutions
should be suitably buffered if necessary and the liquid diluent
first rendered isotonic with sufficient saline or glucose. These
particular aqueous solutions are especially suitable for
intravenous, intramuscular, subcutaneous and intraperitoneal
administration. The oily solutions are suitable for
intra-articular, intra-muscular and subcutaneous injection
purposes.
[0300] In addition to the aforementioned ingredients, the
compositions of a compound of Formula (I) or (II), or a
pharmaceutically acceptable salt thereof, may include one or more
additional ingredients such as diluents, buffers, flavouring
agents, colourant, surface active agents, thickeners,
preservatives, e.g., methyl hydroxybenzoate (including
anti-oxidants), emulsifying agents and the like.
[0301] The term "therapeutically effective amount", as used herein,
refers to an amount of a pharmaceutical agent to treat, ameliorate,
or prevent an identified disease, disorder, or condition, or to
exhibit a detectable therapeutic or inhibitory effect. The effect
can be detected by any assay method known in the art. The precise
effective amount for a subject will depend upon the subject's body
weight, size, and health; the nature and extent of the condition;
and the therapeutic or combination of therapeutics selected for
administration.
[0302] Therapeutically effective amounts for a given situation can
be determined by routine experimentation that is within the skill
and judgment of the clinician. In a preferred aspect, the disease
or disorder to be treated is a disease or disorder associated with
.alpha.-amino-.beta.-carboxymuconate-.epsilon.-semialdehyde
decarboxylase (ACMSD) dysfunction.
[0303] For any compound, the therapeutically effective amount can
be estimated initially either in cell culture assays, e.g., in
cells, or in animal models, usually rats, mice, rabbits, dogs, or
pigs. The animal model may also be used to determine the
appropriate concentration range and route of administration. Such
information can then be used to determine useful doses and routes
for administration in humans. Therapeutic/prophylactic efficacy and
toxicity may be determined by standard pharmaceutical procedures in
cell cultures or experimental animals, e.g., ED.sub.50 (the dose
therapeutically effective in 50% of the population) and LD.sub.50
(the dose lethal to 50% of the population). The dose ratio between
toxic and therapeutic effects is the therapeutic index, and it can
be expressed as the ratio, LD.sub.50/ED.sub.50.
[0304] Pharmaceutical compositions that exhibit large therapeutic
indices are preferred. The dosage may vary within this range
depending upon the dosage form employed, sensitivity of the
patient, and the route of administration.
[0305] Dosage and administration are adjusted to provide sufficient
levels of the active agent(s) or to maintain the desired effect.
Factors which may be taken into account include the severity of the
disease state, general health of the subject, age, weight, and
gender of the subject, diet, time and frequency of administration,
drug combination(s), reaction sensitivities, and tolerance/response
to therapy. Long-acting pharmaceutical compositions may be
administered every 3 to 4 days, every week, or once every two weeks
depending on half-life and clearance rate of the particular
formulation.
[0306] A suitable dosage of the compound of Formula (I) or (II), or
a pharmaceutically acceptable salt thereof, will depend on the age
and condition of the patient, the severity of the disease to be
treated and other factors well known to the practicing physician.
The compound may be administered for example either orally,
parenterally or topically according to different dosing schedules,
e.g., daily or with intervals, such as weekly intervals. In general
a single dose will be in the range from 0.01 to 500 mg/kg body
weight, preferably from about 0.05 to 100 mg/kg body weight, more
preferably between 0.1 to 50 mg/kg body weight, and most preferably
between 0.1 to 25 mg/kg body weight. The compound may be
administered as a bolus (i.e., the entire daily dose is
administered at once) or in divided doses two or more times a day.
Variations based on the aforementioned dosage ranges may be made by
a physician of ordinary skill taking into account known
considerations such as weight, age, and condition of the person
being treated, the severity of the affliction, and the particular
route of administration.
[0307] The compounds of Formula (I) or (II), or a pharmaceutically
acceptable salt thereof, may also be prepared in a pharmaceutical
composition comprising one or more further active substances alone,
or in combination with pharmaceutically acceptable carriers,
diluents, or excipients in either single or multiple doses. The
suitable pharmaceutically acceptable carriers, diluents and
excipients are as described herein above, and the one or more
further active substances may be any active substances, or
preferably an active substance as described in the section
"combination treatment" herein below.
Methods of Treatment
[0308] In another aspect, the present disclosure relates to a
method of preventing, reducing the risk of, or ameliorating a
disease or disorder in which
.alpha.-amino-.beta.-carboxymuconate-.epsilon.-semialdehyde
decarboxylase (ACMSD) plays a role comprising administering to the
subject in need thereof a therapeutically effective amount of one
or more compounds of Formula (I) or (II), or a pharmaceutically
acceptable salt thereof.
[0309] Another aspect of the present disclosure relates to a method
of preventing, reducing the risk of, or ameliorating a disease or
disorder in which
.alpha.-amino-.beta.-carboxymuconate-.epsilon.-semialdehyde
decarboxylase (ACMSD) plays a role comprising administering to the
subject in need thereof a therapeutically effective amount of a
pharmaceutical composition comprising one or more compounds of
Formula (I) or (II), or a pharmaceutically acceptable salt thereof,
and at least one of a pharmaceutically acceptable carrier, diluent,
or excipient.
[0310] Another aspect of the present disclosure relates to a method
of treating, preventing, reducing the risk of, or ameliorating a
disease or disorder by inhibition of
.alpha.-amino-.beta.-carboxymuconate-.epsilon.-semialdehyde
decarboxylase (ACMSD) comprising administering to the subject
suffering from or susceptible to developing the disease or disorder
a therapeutically effective amount of one or more compounds of
Formula (I) or (II), or a pharmaceutically acceptable salt
thereof.
[0311] Another aspect of the present disclosure relates to a method
of treating, preventing, reducing the risk of, or ameliorating a
disease or disorder by inhibition of
.alpha.-amino-.beta.-carboxymuconate-.epsilon.-semialdehyde
decarboxylase (ACMSD) comprising administering to the subject
suffering from or susceptible to developing the disease or disorder
associated with ACMSD a therapeutically effective amount of a
pharmaceutical composition comprising one or more compounds of
Formula (I) or (II), or a pharmaceutically acceptable salt thereof,
and at least one of a pharmaceutically acceptable carrier, diluent,
or excipient.
[0312] In another aspect, the present disclosure relates to a
method of treating, preventing, reducing the risk of, or
ameliorating a disease or disorder in which nicotinamide adenine
dinucleotide (NAD.sup.+) modulation plays a role comprising
administering to the subject in need thereof a therapeutically
effective amount of one or more compounds of Formula (I) or (II),
or a pharmaceutically acceptable salt thereof.
[0313] In another aspect, the present disclosure relates to a
method of treating, preventing, reducing the risk of, or
ameliorating a disease or disorder in which nicotinamide adenine
dinucleotide (NAD.sup.+) modulation plays a role comprising
administering to the subject in need thereof a therapeutically
effective amount of a pharmaceutical composition comprising one or
more compounds of Formula (I) or (II), or a pharmaceutically
acceptable salt thereof, and at least one of a pharmaceutically
acceptable carrier, diluent, or excipient.
[0314] In another aspect, the present disclosure relates to a
method of treating, preventing, reducing the risk of, or
ameliorating a disease or disorder associated with reduced
nicotinamide adenine dinucleotide (NAD.sup.+) levels comprising
administering to the subject suffering from or susceptible to
developing a disease or disorder associated with reduced NAD.sup.+
levels a therapeutically effective amount of one or more compounds
of Formula (I) or (II), or a pharmaceutically acceptable salt
thereof.
[0315] In another aspect, the present disclosure relates to a
method of treating, preventing, reducing the risk of, or
ameliorating a disease or disorder associated with reduced
nicotinamide adenine dinucleotide (NAD.sup.+) levels comprising
administering to the subject suffering from or susceptible to
developing a disease or disorder associated with reduced NAD.sup.+
levels a therapeutically effective amount of a pharmaceutical
composition comprising one or more compounds of Formula (I) or
(II), or a pharmaceutically acceptable salt thereof, and at least
one of a pharmaceutically acceptable carrier, diluent, or
excipient.
[0316] Another aspect of the present disclosure relates to a method
of treating, preventing, reducing the risk of, or ameliorating a
disorder associated with mitochondrial dysfunction comprising
administering to the subject suffering from or susceptible to
developing a metabolic disorder a therapeutically effective amount
of one or more compounds of Formula (I) or (II), or a
pharmaceutically acceptable salt thereof. In one embodiment, the
disorder associated with mitochondrial dysfunction is an inherited
mitochondrial disease, a common metabolic disorder, a
neurodegenerative disease, an aging related disorder, a kidney
disorder, or a chronic inflammatory disease. In a preferred
embodiment, the disorder associated with mitochondrial dysfunction
is a common metabolic disorder such as obesity or type II
diabetes.
[0317] Another aspect of the present disclosure relates to a method
of treating, preventing, reducing the risk of, or ameliorating a
disorder associated with mitochondrial dysfunction comprising
administering to the subject suffering from or susceptible to
developing a metabolic disorder a therapeutically effective amount
of a pharmaceutical composition comprising one or more compounds of
Formula (I) or (II), or a pharmaceutically acceptable salt thereof,
and at least one of a pharmaceutically acceptable carrier, diluent,
or excipient. In one embodiment, the disorder associated with
mitochondrial dysfunction is an inherited mitochondrial disease, a
common metabolic disorder, a neurodegenerative disease, an aging
related disorder, a kidney disorder, or a chronic inflammatory
disease. In a preferred embodiment, the disorder associated with
mitochondrial dysfunction is a common metabolic disorder such as
obesity or type II diabetes.
[0318] In another aspect, the present disclosure relates to a
method of promoting oxidative metabolism comprising administering
to the subject suffering from or susceptible to developing a
metabolic disorder a therapeutically effective amount of one or
more compounds of Formula (I) or (II), or a pharmaceutically
acceptable salt thereof, that increases intracellular nicotinamide
adenine dinucleotide (NAD.sup.+).
[0319] In another aspect, the present disclosure relates to a
method of promoting oxidative metabolism comprising administering
to the subject suffering from or susceptible to developing a
metabolic disorder a therapeutically effective amount of a
pharmaceutical composition comprising one or more compounds of
Formula (I) or (II), or a pharmaceutically acceptable salt thereof,
and at least one of a pharmaceutically acceptable carrier, diluent,
or excipient, that increases intracellular nicotinamide adenine
dinucleotide (NAD.sup.+).
[0320] In yet another aspect, the present disclosure relates to a
method for the manufacture of a medicament for treating,
preventing, reducing the risk of, or ameliorating a disease or
condition that can be mediated by ACMSD inhibition, wherein the
medicament comprises a compound of Formula (I) or (II), or a
pharmaceutically acceptable salt thereof.
[0321] In another aspect, the present disclosure relates to a
method for the manufacture of a medicament for treating,
preventing, reducing the risk of, or ameliorating a disease or
condition that can be mediated by ACMSD inhibition, wherein the
medicament comprises a pharmaceutical composition comprising one or
more compounds of Formula (I) or (II), or a pharmaceutically
acceptable salt thereof, and at least one of a pharmaceutically
acceptable carrier, diluent, or excipient.
[0322] In yet another aspect, the present disclosure relates to a
compound for use in a method for treating, preventing, reducing the
risk of, or ameliorating a disease or condition that can be
mediated by ACMSD inhibition, wherein the compound comprises a
compound of Formula (I) or (II), or a pharmaceutically acceptable
salt thereof.
[0323] In another aspect, the present disclosure relates to a
pharmaceutical composition for use in a method for treating,
preventing, reducing the risk of, or ameliorating a disease or
condition that can be mediated by ACMSD inhibition, wherein the
composition comprises one or more compounds of compound of Formula
(I) or (II), or a pharmaceutically acceptable salt thereof, and at
least one of a pharmaceutically acceptable carrier, diluent, or
excipient.
[0324] Another aspect of the present disclosure relates to the use
of a compound of Formula (I) or (II), or a pharmaceutically
acceptable salt thereof in the manufacture of a medicament for
treating, preventing, reducing the risk of, or ameliorating a
disease or disorder by inhibition of
.alpha.-amino-.beta.-carboxymuconate-.epsilon.-semialdehyde
decarboxylase (ACMSD).
[0325] Another aspect of the present disclosure relates to the use
of a pharmaceutical composition comprising one or more compounds of
Formula (I) or (II), or a pharmaceutically acceptable salt thereof,
and at least one of a pharmaceutically acceptable carrier, diluent,
or excipient, in the manufacture of a medicament for treating,
preventing, reducing the risk of, or ameliorating a disease or
disorder by inhibition of
.alpha.-amino-.beta.-carboxymuconate-s-semialdehyde decarboxylase
(ACMSD).
[0326] In another aspect, the present disclosure relates to the use
of a compound of a compound of Formula (I) or (II), or a
pharmaceutically acceptable salt thereof in the manufacture of a
medicament for treating, preventing, reducing the risk of, or
ameliorating a disease or disorder associated with reduced
nicotinamide adenine dinucleotide (NAD.sup.+) levels.
[0327] In another aspect, the present disclosure relates to the use
of a pharmaceutical composition comprising one or more compounds of
Formula (I) or (II), or a pharmaceutically acceptable salt thereof,
and at least one of a pharmaceutically acceptable carrier, diluent,
or excipient, in the manufacture of a medicament for treating,
preventing, reducing the risk of, or ameliorating a disease or
disorder associated with reduced nicotinamide adenine dinucleotide
(NAD.sup.+) levels.
[0328] Another aspect of the present disclosure relates to the use
of a compound of Formula (I) or (II), or a pharmaceutically
acceptable salt thereof in the manufacture of a medicament for
treating, preventing, reducing the risk of, or ameliorating a
disorder associated with mitochondrial dysfunction.
[0329] Another aspect of the present disclosure relates to the use
of a pharmaceutical composition comprising one or more compounds of
Formula (I) or (II), or a pharmaceutically acceptable salt thereof,
and at least one of a pharmaceutically acceptable carrier, diluent,
or excipient, in the manufacture of a medicament for treating,
preventing, reducing the risk of, or ameliorating a disorder
associated with mitochondrial dysfunction.
[0330] In another aspect, the present disclosure relates to the use
of a compound of a compound of Formula (I) or (II), or a
pharmaceutically acceptable salt thereof, in the manufacture of a
medicament for promoting oxidative metabolism.
[0331] In another aspect, the present disclosure relates to the use
of a pharmaceutical composition comprising one or more compounds of
Formula (I) or (II), or a pharmaceutically acceptable salt thereof,
and at least one of a pharmaceutically acceptable carrier, diluent,
or excipient, in the manufacture of a medicament for promoting
oxidative metabolism.
[0332] Another aspect of the present disclosure relates to a
compound of Formula (I) or (II), or a pharmaceutically acceptable
salt thereof, for use in the manufacture of a medicament for
treating, preventing, reducing the risk of, or ameliorating a
disease or disorder by inhibition of
.alpha.-amino-.beta.-carboxymuconate-.epsilon.-semialdehyde
decarboxylase (ACMSD).
[0333] Another aspect of the present disclosure relates to a
pharmaceutical composition comprising one or more compounds of
Formula (I) or (II), or a pharmaceutically acceptable salt thereof,
and at least one of a pharmaceutically acceptable carrier, diluent,
or excipient, for use in the manufacture of a medicament for
treating, preventing, reducing the risk of, or ameliorating a
disease or disorder by inhibition of
.alpha.-amino-.beta.-carboxymuconate-.epsilon.-semialdehyde
decarboxylase (ACMSD).
[0334] In another aspect, the present disclosure relates to a
compound of Formula (I) or (II), or a pharmaceutically acceptable
salt thereof for use as a medicament for treating, preventing,
reducing the risk of, or ameliorating a disease or disorder
associated with reduced nicotinamide adenine dinucleotide
(NAD.sup.+) levels.
[0335] In another aspect, the present disclosure relates to a
pharmaceutical composition comprising one or more compounds of
Formula (I) or (II), or a pharmaceutically acceptable salt thereof,
and at least one of a pharmaceutically acceptable carrier, diluent,
or excipient, for use as a medicament for treating, preventing,
reducing the risk of, or ameliorating a disease or disorder
associated with reduced nicotinamide adenine dinucleotide
(NAD.sup.+) levels.
[0336] Another aspect of the present disclosure relates to a
compound of Formula (I) or (II), or a pharmaceutically acceptable
salt thereof for use as a medicament for treating, preventing,
reducing the risk of, or ameliorating a disorder associated with
mitochondrial dysfunction.
[0337] Another aspect of the present disclosure relates to a
pharmaceutical composition comprising one or more compounds of
Formula (I) or (II), or a pharmaceutically acceptable salt thereof,
and at least one of a pharmaceutically acceptable carrier, diluent,
or excipient, for use as a medicament for treating, preventing,
reducing the risk of, or ameliorating a disorder associated with
mitochondrial dysfunction.
[0338] In another aspect, the present disclosure relates to a
compound of Formula (I) or (II), or a pharmaceutically acceptable
salt thereof for use as a medicament for promoting oxidative
metabolism.
[0339] In another aspect, the present disclosure relates to a
pharmaceutical composition comprising one or more compounds of
Formula (I) or (II), or a pharmaceutically acceptable salt thereof,
and at least one of a pharmaceutically acceptable carrier, diluent,
or excipient, for use as a medicament for promoting oxidative
metabolism.
[0340] Another aspect of the present disclosure relates to a
compound of Formula (I) or (II), or a pharmaceutically acceptable
salt thereof for use in treating, preventing, reducing the risk of,
or ameliorating a disease or disorder associated with reduced
nicotinamide adenine dinucleotide (NAD.sup.+) levels.
[0341] Another aspect of the present disclosure relates to a
pharmaceutical composition comprising one or more compounds of
Formula (I) or (II), or a pharmaceutically acceptable salt thereof,
and at least one of a pharmaceutically acceptable carrier, diluent,
or excipient, for use in treating, preventing, reducing the risk
of, or ameliorating a disease or disorder associated with reduced
nicotinamide adenine dinucleotide (NAD.sup.+) levels.
[0342] In another aspect, the present disclosure relates to a
compound of Formula (I) or (II), or a pharmaceutically acceptable
salt thereof for use in for treating, preventing, reducing the risk
of, or ameliorating a disorder associated with mitochondrial
dysfunction.
[0343] In another aspect, the present disclosure relates to a
pharmaceutical composition comprising one or more compounds of
Formula (I) or (II), or a pharmaceutically acceptable salt thereof,
and at least one of a pharmaceutically acceptable carrier, diluent,
or excipient, for use in for treating, preventing, reducing the
risk of, or ameliorating a disorder associated with mitochondrial
dysfunction.
[0344] Another aspect of the present disclosure relates to a
compound of Formula (I) or (II), or a pharmaceutically acceptable
salt thereof for use in promoting oxidative metabolism.
[0345] Another aspect of the present disclosure relates to a
pharmaceutical composition comprising one or more compounds of
Formula (I) or (II), or a pharmaceutically acceptable salt thereof,
and at least one of a pharmaceutically acceptable carrier, diluent,
or excipient, for use in promoting oxidative metabolism.
[0346] In some embodiments, the disease or disorder associate with
reduced nicotinamide adenine dinucleotide (NAD.sup.+) levels is a
chronic liver disease including, but is not limited to, primary
biliary cirrhosis (PBC), cerebrotendinous xanthomatosis (CTX),
primary sclerosing cholangitis (PSC), drug induced cholestasis,
intrahepatic cholestasis of pregnancy, parenteral nutrition
associated cholestasis (PNAC), bacterial overgrowth or sepsis
associated cholestasis, autoimmune hepatitis, chronic viral
hepatitis, alcoholic liver disease, nonalcoholic fatty liver
disease (NAFLD), nonalcoholic steatohepatitis (NASH), liver
transplant associated graft versus host disease, living donor
transplant liver regeneration, congenital hepatic fibrosis,
choledocholithiasis, granulomatous liver disease, intra- or
extrahepatic malignancy, Sjogren's syndrome, Sarcoidosis, Wilson's
disease, Gaucher's disease, hemochromatosis, and alpha
1-antitrypsin deficiency. In one embodiment, the common metabolic
disorder is obesity or type II diabetes.
[0347] In some embodiments, the disorder associated with
mitochondrial dysfunction is an inherited mitochondrial disease, a
common metabolic disorder, a neurodegenerative disease, an aging
related disorder, a kidney disorder, or a chronic inflammatory
disease.
[0348] In another aspect, the present disclosure relates to a
method of treating, preventing, reducing the risk of, or
ameliorating a disease or disorder by inhibition of
.alpha.-amino-.beta.-carboxymuconate-.epsilon.-semialdehyde
decarboxylase (ACMSD), comprising administering to a subject in
need thereof, a therapeutically effective amount of a compound of
Formula (I) or (II) or a pharmaceutical composition comprising a
compound of Formula (I) or (II).
[0349] As used herein, "treating" or "treat" describes the
management and care of a patient for the purpose of reversing,
inhibiting, or combating a disease, condition, or disorder and
includes the administration of a compound of the present disclosure
(i.e., a compound of Formula (I) or (II)), or a pharmaceutically
acceptable salt, prodrug, metabolite, polymorph or solvate thereof,
to reverse the disease, condition, or disorder, eliminate the
disease, condition, or disorder, or inhibit the process of the
disease, condition, or disorder.
[0350] A compound of the present disclosure (i.e., a compound of
Formula (I) or (II)), or a pharmaceutically acceptable salt,
prodrug, metabolite, polymorph or solvate thereof, can also be used
to prevent a disease, condition, or disorder or one or more
symptoms of such disease, condition, or disorder. As used herein,
"preventing" or "prevent" describes reducing or eliminating the
onset of the symptoms or complications of the disease, condition,
or disorder.
[0351] A compound of the present disclosure (i.e., a compound of
Formula (I) or (II)), or a pharmaceutically acceptable salt,
prodrug, metabolite, polymorph or solvate thereof, can also be used
to alleviate one or more symptoms of such disease, condition, or
disorder. As used herein, the term "alleviate" is meant to describe
a process by which the severity of a sign or symptom of a disorder
is decreased. Importantly, a sign or symptom can be alleviated
without being eliminated. Preferably treatment is curative or
ameliorating.
Clinical Conditions and Other Uses of Compounds
[0352] The compounds according to Formula (I) or (II), or a
pharmaceutically acceptable form thereof, compositions,
medicaments, and compounds for use, as defined herein, are useful
for treatment of a disease or disorder in which
.alpha.-amino-.beta.-carboxymuconate-.epsilon.-semialdehyde
decarboxylase (ACMSD) modulation plays a role. The compounds may be
used either in human or in veterinary medicine and the patient may
be any mammal, but especially a human. The treatment may include
administering to any mammal, but especially a human, suffering from
a disease or disorder in which
.alpha.-amino-.beta.-carboxymuconate-.epsilon.-semialdehyde
decarboxylase (ACMSD) modulation plays a role, a therapeutically
effective amount of a compound according to Formula (I) or (II), or
a pharmaceutically acceptable salt thereof, as defined herein.
[0353] The present disclosure also relates to a compound of Formula
(I) or (II), or a pharmaceutically acceptable salt thereof, as
defined herein, for use in a disease or disorder associated with
.alpha.-amino-.beta.-carboxymuconate-.epsilon.-semialdehyde
decarboxylase (ACMSD) dysfunction, such as obesity, type II
diabetes and its complications (e.g., diabetic retinopathy and
nephropathy), non-alcoholic fatty liver disease (NAFLD),
non-alcoholic steatohepatitis (NASH), or chronic kidney
disease.
[0354] By the term "disease or disorder associated with
.alpha.-amino-.beta.-carboxymuconate-.epsilon.-semialdehyde
decarboxylase (ACMSD) dysfunction" is meant any disease
characterized by reduced nicotinamide adenine dinucleotide (NAD)
expression and/or activity in at least in some instances of the
disease, or a disease which is ameliorated by elevation of the
levels of NAD.sup.+.
[0355] The methods, medicaments and compounds for use of the
present disclosure are useful to treat, alleviate the symptoms of,
or delay the onset of a disorder associated with aberrant
mitochondrial function. Disorders associated with aberrant
mitochondrial function include, for example, metabolic disorders,
neurodegenerative disorders, aging related disorders, and chronic
inflammatory disorders. Mitochondrial disorders also include
diseases with inherited and/or acquired mitochondrial dysfunction
(i.e., Charcot-Marie-Tooth disease, Type 2A2, Mitochondrial
Encephalopathy Lactic Acidosis and Stroke (MELAS), Leigh syndrome,
Barth syndrome, and Leber's optic neuropathy), fatty acid oxidation
disorders, inherited forms of deafness and blindness, and metabolic
abnormalities induced by exposure to toxic chemicals and/or drugs
(e.g., cisplatin induced deafness).
[0356] Metabolic disorders include, for example, type II diabetes,
obesity, hyperglycemia, glucose intolerance, insulin resistance
(i.e., hyperinsulinemia, metabolic syndrome, syndrome X),
hypercholesterolemia, hypertension, hyperlipoproteinemia,
hyperlipidemia (e.g., dyslipidemia), hypertriglylceridemia,
cardiovascular disease, atherosclerosis, peripheral vascular
disease, kidney disease, ketoacidosis, thrombotic disorders,
nephropathy, diabetic neuropathy, diabetic retinopathy, sexual
dysfunction, dermatopathy, dyspepsia, hypoglycemia, cancer, and
edema.
[0357] Neurodegenerative disorders include diseases such as
photoreceptor degeneration (i.e., retinitis pigmentosa), Dementia,
Alzheimer's disease, Parkinson's disease, and Huntington's
disease.
[0358] Chronic inflammatory diseases include diseases such as
celiac disease, vasculitis, lupus, chronic obstructive pulmonary
disease (COPD), irritable bowel disease, atherosclerosis,
arthritis, and psoriasis.
[0359] Aging related disorders include diseases such as cancer,
dementia, cardiovascular disease (i.e., arteriosclerosis),
hypertension, diabetes mellitus (type I or type II), arthritis,
cataracts, Alzheimer's disease, macular degeneration, and
osteoporosis.
[0360] The subject can be suffering from or susceptible to
developing a metabolic disorder. Subjects suffering from or at risk
of developing a metabolic disorder are identified by methods known
in the art. For example, diabetes can be diagnosed by measuring
fasting blood glucose levels or insulin or by glucose tolerance
test. Normal adult glucose levels are between about 60-126 mg/dl.
Normal insulin levels are about 7 mU/mL.+-.3 mU.
[0361] Hypertension can be diagnosed by a blood pressure reading
consistently at or above about 140/90. Cardiovascular disease can
be diagnosed by measuring cholesterol levels. For example, LDL
cholesterol above about 137 or total cholesterol above about 200 is
indicative of cardiovascular disease. Hyperglycemia can be
diagnosed by a blood glucose level higher than about 10 mmol/l (180
mg/dl). Glucose intolerance can be diagnosed by glucose levels of
140 to 199 mg per dL (7.8 to 11.0 mmol) after conducting a 75 g
oral two-hour glucose tolerance test. Insulin resistance can be
diagnosed by a fasting serum insulin level of greater than
approximately 60 pmol/L. Hypoglycemia can be diagnosed by a blood
glucose level lower than about 2.8 to 3.0 mmol/L (50 to 54 mg/dl).
Obesity can be diagnosed, for example, by body mass index. Body
mass index (BMI) is measured in kg/m.sup.2 (or
lb/in.sup.2.times.704.5).
[0362] Alternatively, waist circumference (estimates fat
distribution), waist-to-hip ratio (estimates fat distribution),
skinfold thickness (if measured at several sites, estimates fat
distribution), or bioimpedance (based on principle that lean mass
conducts current better than fat mass (i.e., fat mass impedes
current), estimates % fat) can be measured. The parameters for
normal, overweight, or obese individuals are as follows:
Underweight: BMI<18.5; Normal: BMI about 18.5 to about 24.9;
Overweight: BMI=about 25 to about 29.9. Overweight individuals are
characterized as having a waist circumference of >94 cm for men
or >80 cm for women and waist to hip ratios of .gtoreq.0.95 in
men and .gtoreq.0.80 in women. Obese individuals are characterized
as having a BMI of 30 to 34.9, being greater than 20% above
"normal" weight for height, having a body fat percentage >30%
for women and 25% for men, and having a waist circumference >102
cm (40 inches) for men or 88 cm (35 inches) for women. Individuals
with severe or morbid obesity are characterized as having a BMI of
.gtoreq.35.
[0363] The methods described herein may lead to a reduction in the
severity or the alleviation of one or more symptoms of a metabolic
disorder. For example, symptoms of diabetes include elevated
fasting blood glucose levels, blood pressure at or above 140/90
mm/Hg; abnormal blood fat levels, such as high-density lipoproteins
(HDL) less than or equal to 35 mg/dL, or triglycerides greater than
or equal to 250 mg/dL (mg/dL=milligrams of glucose per deciliter of
blood). Efficacy of treatment is determined in association with any
known method for diagnosing the metabolic disorder. Alleviation of
one or more symptoms of the metabolic disorder indicates that the
compound confers a clinical benefit.
[0364] The methods of the present disclosure are useful to treat,
alleviate the symptoms of, or delay the onset of a kidney disorder.
Kidney disorders include acute kidney injury (AKI) and chronic
kidney disease (CKD).
[0365] The subject can be suffering from or susceptible to
developing acute kidney injury (AKI). The acute kidney injury can
be characterized by one or more clinical criteria or conditions
(i.e., an abrupt decrease in the ability of the kidneys to excrete
nitrogenous waste products from the blood, resulting in azotemia).
Subjects suffering from or at risk of developing acute kidney
injury (AKI) are identified by methods known in the art. For
example, the acute kidney injury can be characterized by an
increase in serum creatinine by at least 50% over baseline, an
absolute increase in serum creatinine of at least 0.3 mg/dL over
baseline, a reduction in glomerular filtration rate of at least 25%
compared to baseline, a decrease in urine output to 0.5 ml per
kilogram of body weight or less per hour persisting for at least 6
hours, or any combination thereof. An acute kidney injury may be
caused by ischemia, drugs or toxic agents (i.e., radiocontrast
media, a non-steroidal anti-inflammatory drug (NSAID), alcohol, or
a chemotherapy agent), viruses, and obstruction.
[0366] The subject can be suffering from or susceptible to
developing chronic kidney disease (CKD). Chronic kidney disease
(CKD) is defined as either (1) having kidney damage as defined by
structural or functional abnormalities of the kidney for 3 months
or longer with or without a decreased glomerular filtration rate
(GFR) or (2) having a GFR of less than 60 mL/min/1.73 m.sup.2 for 3
months or longer with or without kidney damage. Subjects suffering
from or at risk of developing a chronic kidney disease (CKD) are
identified by methods known in the art. Structural or functional
abnormalities are manifested by symptoms such as either pathologic
abnormalities or markers of kidney damage, including abnormalities
identified in imaging studies or the composition of blood or
urine.
[0367] For example, CKD can be diagnosed by testing for specific
marker. For example, markers of kidney damage include a plasma
creatinine concentration of above about 1.6 mg/dL and a blood urea
nitrogen (BUN) concentration of above about 20 mg/dL. Typically,
both of these markers are elevated in individuals with CKD.
Additional markers of kidney damage can include hematuria (i.e.,
any detectable amount of blood in the urine), proteinuria (i.e.,
protein concentrations in urine above about 100 mg/dL), albuminuria
(i.e., albumin concentrations in urine above about 100 mg/dL), an
intact parathyroid hormone (PTH) concentration in the blood above
about 150 pg/mL, or blood phosphate levels of above about 4.5
mg/dL. One specific marker of kidney disease is a GFR rate above
normal (i.e., a GFR above about 90 mL/min/1.73 m.sup.2), however a
below normal GFR also indicates CKD.
[0368] The methods of the present disclosure are useful to treat,
alleviate the symptoms of, or delay the onset of non-alcoholic
fatty liver disease (NAFLD) and/or non-alcoholic steatohepatitis
(NASH). The subject can be suffering from or susceptible to
developing non-alcoholic fatty liver disease (NAFLD) and/or
non-alcoholic steatohepatitis (NASH). Subjects suffering from or at
risk of developing a non-alcoholic fatty liver disease (NAFLD)
and/or non-alcoholic steatohepatitis (NASH) are identified by
methods known in the art. For example, NAFLD and/or NASH can be
diagnosed by liver biopsy.
[0369] Non-alcoholic fatty liver disease (NAFLD), as defined
herein, is a disease with fat deposition in the liver, which occurs
in patients whose alcohol ingestion history is not long enough to
cause liver injury. Non-alcoholic fatty liver disease (NAFLD) can
be further classified into simple fatty liver, steatohepatitis and
cirrhosis. Nonalcoholic steatohepatitis (NASH) refers to a
pathology associated with inflammation, liver cell necrosis,
ballooning and fibrosis. The onset of nonalcoholic simple fatty
liver is induced by fat deposition in liver cells, and this fat
accumulation is defined by the balance between increasing factors
(influx and synthesis of fats in liver cells) and decreasing
factors (catabolism of fats and their release from liver cells).
Once damage of liver cells occurs, in addition to this fat
deposition, nonalcoholic simple fatty liver will progress to
nonalcoholic steatohepatitis. Nonalcoholic steatohepatitis is
progressive and may finally progress to cirrhosis and
hepatocellular carcinoma.
Combination Treatment
[0370] In another aspect, the disclosure includes a compound of
Formula (I) or (II), or a pharmaceutically acceptable salt thereof,
for use in a combination therapy. A compound, compositions,
medicaments and compounds for use of Formula (I) or (II), or a
pharmaceutically acceptable salt thereof, may also be used to
advantage in combination with one or more other therapeutic agents.
Such therapeutic agents include, but are not limited to other ACMSD
inhibitors; anti-diabetic agents such as PPARy agonists,
PPAR.alpha./.gamma. dual agonists, PPARS agonists, biguanides,
protein tyrosine phosphatase-1B (PTP-1B), dipeptidyl peptidase IV
(DPP-IV) inhibitors, sulfonylureas, meglitinides, alpha glucoside
hydrolase inhibitors, alpha-amylase inhibitors, insulin
secreatagogues, A2 antagonists, insulin or insulin mimetics,
glycogen phosphorylase inhibitors, GLP-1 agonists,
non-thiazolidinediones, glycokinase, and 11 HSD-1 inhibitor;
anti-obesity agents such as uncoupling Protein (UCP-1, UCP-2, and
UCP-3) activators, .beta.3 adrenergic receptor (.beta.3), thyroid
hormone p agonists, fatty acid synthase (PAS) inhibitors,
phosphodieterase (PDE) inhibitors, lipase inhibitors, serotonin
reuptake inhibitors, monoamine reuptake inhibitors, Mc4r agonists,
5HT2c agonists, growth hormone secretagogue (GHS) agonists, CNTF
derivatives, ciliary neurotrophic factors (CNTh), cholecystokinin-A
(CCK-A) agonists, opioid antagonists, orexin antagonists,
acyl-estrogens, leptin, NPY 5 antagonists, neuropeptide Y5 (NPY5)
antagonists, neuropeptide Y2 (NPY2) agonists, melanin-concentrating
hormone receptor (MCHLR) antagonists and melanin-concentrating
hormone 2 receptor (MCH2R), MCH1R antagonists, neuropeptide Y1,
ghrelin antagonists, cannabinoid receptor 1 (CB-1), serotonin (5HT)
transport inhibitors, CCK-A agonists and histamine 3 (H3)
antagonist/inverse agonists; cholesterol lower agents such as
3-hydroxy-3-methylglutaiyl-coenzyme A (HMG CoA) reductase
inhibitors, HMG-CoA synthase inhibitors, squalene epoxidase
inhibitors, fibric acids, bile acid-binding resins probucol and
niacin (nicotinic acid); compounds that boost NAD.sup.+ levels such
as NAD.sup.+ precursors (i.e. nicotinamide ribose (NA),
nicotinamide mononucleotide (NMN), nicotinic acid (NA) and
nicotinamide); and compounds that inhibit NAD.sup.+ consumption
such as PARP inhibitors and CD38 inhibitors.
[0371] PPARy agonists useful in the present disclosure include, but
are not limited to, glitazones (e.g., balaglitazone, ciglitazone,
darglitazone, englitazone, isaglitazone (MCC-555), pioglitazone,
rosiglitazone, troglitazone, CLX-0921, 5-BTZD, and the like);
GW-0207, LG-100641, LY-300512, LY-519818, R483 (Roche), T131
(Tularik), and compounds disclosed in WO97/27857, 97/28115,
97/28137 and 97/27847; and pharmaceutically acceptable salts or
esters thereof. PPAR.alpha./.gamma. dual agonists useful in the
present disclosure, include, but are not limited to, CLX-0940,
GW-1536, GW1929, GW-2433, KRP-297, L-796449, LR-90, MK-0767, SB
219994, and muraglitazar, and pharmaceutically acceptable salts or
esters thereof KRP-297 is
5-[(2,4-Dioxo-5-thiazolidinyl)methyl]-2-methoxy-N-[[4-(trifluoromethyl)
phenyl] methyl]benzamide, and pharmaceutically acceptable salts or
esters thereof. PPAR.delta. agonists useful in the present
disclosure include, but are not limited to, GW 501516, GW 590735,
and compounds disclosed in JP 10237049, WO 02/14291, and WO
2018/125983; and pharmaceutically acceptable salts or esters
thereof.
[0372] Biguanides useful in the present disclosure include, but are
not limited to, buformin, metformin, and phenformin, and
pharmaceutically acceptable salts or esters thereof. Metformin
(Glucophage.RTM.) is indicated for patients with non-insulin
dependent diabetes mellitus, particularly those with refractory
obesity. Physician's Desk Reference.RTM. page 1080-1086, (56th ed.
2002).
[0373] Protein tyrosine phosphatase-1B (PTP-1B) inhibitors useful
in the present disclosure include, but are not limited to,
A-401,674, KR 61639, OC-060062, OC-83839, OC-297962, MC52445,
MC52453, and the compounds disclosed in WO 02/26707, WO 02/26743,
JP 2002114768, and pharmaceutically acceptable salts or esters
thereof.
[0374] Dipeptidyl peptidase IV (DPP-IV) inhibitors, such as
isoleucine thiazolidide; NVP-DPP728; P32/98; and LAP 237, P 3298,
TSL 225, valine pyrrolidide, TMC-2A/2B/2C, CD-26 inhibitors, FE
999011, P9310/K364, VIP 0177, DPP4, SDZ 274A444; and the compounds
disclosed in WO 03/00449; WO 03/004496; EP 1 258 476; WO 02/083128;
WO 021062764; WO 03/000250; WO 03/002530; WO 03/002531; WO
03/002553; WO 03/002593; WO 03/000180; and WO 03/000181.
[0375] Sulfonylureas useful in the present disclosure include, but
are not limited to, acetohexamide, chloropropamide, diabinese,
glibenclamide, glipizide, glyburide, glimepiride, gliclazide,
glipentide, gliquidone, glisolamide, tolazamide, and tolbutamide,
pharmaceutically acceptable salts or esters thereof. Meglitinides
useful in the present disclosure include, but are not limited to,
repaglinide and nateglinide, and pharmaceutically acceptable salts
or esters thereof.
[0376] Alpha glucoside hydrolase inhibitors (or glucoside
inhibitors) useful in the present disclosure include, but are not
limited to, acarbose, adiposine, camiglibose, emiglitate, miglitol,
voglibose, pradimicin-Q, salbostatin, CKD-711, MDL-25,637,
MDL-73,945, and MOR 14, and pharmaceutically acceptable salts or
esters thereof, and the compounds disclosed in U.S. Pat. Nos.
4,062,950, 4,174,439, 4,254,256, 4,701,559, 4,639,436, 5,192,772,
4,634,765, 5,157,116, 5,504,078, 5,091,418, 5,217,877, and
5,091,524. Alpha-amylase inhibitors useful in the present
disclosure include, but are not limited to, tendamistat, trestatin,
and A1-3688, and pharmaceutically acceptable salts and esters
thereof, and the compounds disclosed in U.S. Pat. Nos. 4,451,455,
4,623,714, and 4,273,765.
[0377] Insulin secreatagogues useful in the present disclosure
include, but are not limited to, linogliride and A-4166, and
pharmaceutically acceptable salts and esters thereof.
[0378] Fatty acid oxidation inhibitors useful in the present
disclosure include, but are not limited to, clomoxir, and etomoxir,
and pharmaceutically acceptable salts and esters thereof. A2
antagonists useful in the present disclosure include, but are not
`limited to, midaglizole, isaglidole, deriglidole, idazoxan,
earoxan, fluparoxan, and pharmaceutically acceptable salts and
esters thereof. Insulin or insulin mimetics useful in the present
disclosure include, but are not limited to, biota, LP-100,
novarapid, insulin detemir, insulin lispro, insulin glargine,
insulin zinc suspension (lente and ultralente), Lys-Pro insulin,
GLP-1 (73-7) (insulintropin), and GLP-1 (7-36)-NH2), and
pharmaceutically acceptable salts or esters thereof.
[0379] Glycogen phosphorylase inhibitors useful in the present
disclosure include, but are not limited to, CP-368, 296,
CP-316,819, BAYR3401, and compounds disclosed in WO 01/94300, and
WO 02/20530, and pharmaceutically acceptable salts or esters
thereof. GLP-1 agonists useful in the present disclosure include,
but are not limited to, exendin-3 and exendin-4, and compounds
disclosed in US 2003087821 and NZ 504256, and pharmaceutically
acceptable salts or esters thereof.
[0380] Non-thiazolidinediones useful in the present disclosure
include, but are not limited to, JT-501, and farglitazar
(GW-2570/GI-262579), and pharmaceutically acceptable salts or
esters thereof. Glycokinase activators useful in this disclosure,
include, but are not limited to, fused heteroaromatic compounds
such as those disclosed in US 2002103199, and
isoindolin-1-one-substituted propionamide compounds such as those
disclosed in WO 02/48106.
[0381] Serotonin (5HT) transport inhibitors useful in this
disclosure include, but are not limited to, paroxetine, fluoxetine,
fenfluramine, fluvoxamine, sertraline, and imipramine.
Norepinephrine (NE) transport inhibitors useful in this disclosure
include, but are not limited to, GW 320659, despiramine, talsupram,
and nomifensine. Cannabinoid receptor 1 (CB-1) antagonist/inverse
agonists useful in the present disclosure include: U.S. Pat. Nos.
5,532,237, 4,973,587, 5,013,837, 5,081,122, 5,112,820, 5,292,736,
5,624,941 and 6,028,084, and PCT Application Nos. WO 96/33159, WO
98/33765, WO98/43636, WO98/43635, WO 01/09120, WO 98/31227, WO
98/41519, WO 98/37061, WO 00/10967, WO 00/10968, WO 97/29079, WO
99/02499, WO 01/58869, WO 02/076949, WO 01/64632, WO 01/64633, WO
01/64634, and WO 03/007887, and EPO Application No. EP-658546.
Specific CB-1 antagonists/inverse agonists useful in the present
disclosure include, but are not limited to, rimonabant (Sanofi
Synthelabo), SR-147778 (Sanofi Synthelabo), BAY 65-2520 (Bayer),
and SLY 319 (Solvay). CCK-A agonists useful in the present
disclosure include GI 181771, and SR 146,131. Ghrelin antagonists
useful in the present disclosure, include: PCT Application Nos. WO
01/87335, and WO 02/08250. Histamine 3 (H3) antagonist/inverse
agonists useful in the present disclosure include: PCT Application
No. WO 02/15905, and O-[3-(1H-imidazol4-yl)propanol]carbamates
(Kiec-Kononowicz, K. et al., Pharmazie, 55:349-55 (2000)),
piperidine-containing histamine H3-receptor antagonists (Lazewska,
D. et al., Pharmazie, 56:927-32 (2001), benzophenone derivatives
and related compounds (Sasse, A. et al. Arch. Pharm. (Weinheim)
334:45-52 (2001)), substituted N-phenyl carbamates (Reidemeister,
S. et al., Pharmazie, 55:83-6 (2000)), and proxifan derivatives
(Sasse, A. et al., J. Med. Chem. 43:3335-43 (2000)). Specific H3
antagonists/inverse agonists useful in the present disclosure
include, but are not limited to, thioperamide,
3-(1H-imidazol-4-yl)propyl N-4-pentenyl)carbamate, clobenpropit,
iodophenpropit, imoproxifan, GT2394 (Gliatech), and A331440.
[0382] Melanin-concentrating hormone receptor (MCHLR) antagonists
and melanin-concentrating hormone 2 receptor (MCH2R)
agonist/antagonists useful in the present disclosure include PCT
Patent Application Nos. WO 01/82925, WO 01/87834, WO 02/06245, WO
02/04433, and WO 02/51809, and Japanese Patent Application No. JP
13226269. Specific MCH1R antagonists useful in the present
disclosure include, but are not limited to, T-226296 (Takeda), SB
568849, and SNAP 7941. Neuropeptide Y1 (NPY1) antagonists useful in
the present disclosure, include: U.S. Pat. No. 6,001,836, and PCT
Application Nos. WO 96/14307, WO 01/23387, WO 99/51600, WO
01/85690, WO 01/85098, WO 01/85173, and WO 01/89528. Specific
examples of NPY1 antagonists useful in the present disclosure
include, but are not limited to, BIBP3226, J-115814, BIBO 3304,
LY-357897, CP-671906, and GI-264879A. Neuropeptide Y2 (NPY2)
agonists useful in the present disclosure, include, but are not
limited to, peptide YY (PYY), and PYY3_36, peptide YY analogs, PYY
agonists, and the compounds disclosed in WO 03/026591, WO
03/057235, and WO 03/027637. Neuropeptide Y5 (NPY5) antagonists
useful in the present disclosure, include, but are not limited to,
the compounds described in: U.S. Pat. Nos. 6,140,354, 6,191,160,
6,258,837, 6,313,298, 6,337,332, 6,329,395, and 6,340,683,
6,326,375, 6,329,395, 6,337,332, 6,335,345, European Patent Nos.
EP-01010691, and EP 01044970, and PCT-International Patent
Publication Nos. WO 97/19682, WO 97/20820, WO 97/20821, WO
97/20822, WO 97/20823, WO 98/27063, WO 00/107409, WO00/185714, WO
00/185730, WO 00/64880, WO 00/68197, WO 00/69849, wo 01/09120, wo
01/85714, WO 01/85730, WO 01/07409, WO 01/02379, WO 01/02379, WO
01/23388, WO 01/23389, WO 01/44201, WO 01/62737, WO 01/62738, WO
01/09120, WO 02/20488, WO 02/22592, WO 02/48152, WO 02/49648, and
WO 01/14376. Specific NPY5 antagonists useful in the combinations
of the present disclosure, include, but are not limited to
GW-569180A, GW-594884A, GW-587081X, GW-548118X, FR 235,208,
FR226928, FR 240662, FR252384, 1229U91, GI-264879A, CGP71683A,
LY-377897, LY366377, PD-160170, SR-120562A, SR-120819A, JCF-104,
and H409/22. Additional specific NPY5 antagonists useful in the
combinations of the present disclosure, include, but are not
limited to the compounds described in Norman et al., J. Med. Chem.
43:42884312 (2000). Leptin includes, but is not limited to,
recombinant human leptin (PEG-OB, Hoffman La Roche) and recombinant
methionyl human leptin (Amgen). Leptin derivatives (e.g., truncated
forms of leptin) useful in the present disclosure include: U.S.
Pat. Nos. 5,552,524, 5,552,523, 5,552,522, 5,521,283, and PCT
International Publication Nos. WO 96/23513, WO 96/23514, WO
96/23515, WO 96/23516, WO 96/23517, WO 96/23518, WO 96/23519, and
WO 96/23520.
[0383] Opioid antagonists useful in the present disclosure include:
PCT Application No. WO 00/21509. Specific opioid antagonists useful
in the present disclosure include, but are not limited to,
nalmefene (Revex.RTM.), 3-methoxynaltrexone, naloxone, and
naltrexone. Orexin antagonists useful in the present disclosure
include: PCT Patent Application Nos. WO 01/96302, WO 01/68609, WO
02/51232, WO 02/51838, and WO 03/023561. Specific orexin
antagonists useful in the present disclosure include, but are not
limited to, SB-334867-A. Acyl-estrogens useful in the present
disclosure include oleoyl-estrone (del Mar-Grasa, M. et al.,
Obesity Research, 9:202-9 (2001)). Cholecystokinin-A (CCK-A)
agonists useful in the present disclosure include U.S. Pat. No.
5,739,106. Specific CCK-A agonists include, but are not limited to,
AR-R 15849, GI181771, JMv-180, A-71378, A-71623 and SR146131.
Specific ciliary neurotrophic factors (CNTh) useful in the present
disclosure include, but are not limited to, GI-181771
(GlaxoSmithKline), SR146131 (Sanofi Synthelabo), butabindide,
PD170,292, PD 149164 (Pfizer). CNTF derivatives useful in the
present disclosure include, but are not limited to, axokine
(Regeneron), and PCT Application Nos. WO 94/09134, WO 98/22128, and
WO 99/43813. Growth hormone secretagogue (GHS) agonists useful in
the present disclosure include: U.S. Pat. No. 6,358,951, and U.S.
Patent Application Nos. 2002/049196 and 2002/022637, and PCT
Application Nos. WO 01/56592, and WO 02/32888. Specific GHS
agonists include, but are not limited to, NN703, hexarelin,
MK-0677, SM-130686, CP424 391, L-692,429 and L-163,255.
[0384] 5HT2c agonists useful in the present disclosure include:
U.S. Pat. No. 3,914,250, and PCT Application Nos. WO 02/36596, WO
02/48124, WO 02/10169, WO 01/66548, WO 02/44152, WO 02/51844, WO
02/40456, and WO 02/40457. Specific 5HT2c agonists useful in this
disclosure include, but are not limited to, BVT933, DPCA37215,
1K264, PNU 22394, WAY161503, R-1065, and YM 348.
[0385] Mc4r agonists useful in the present disclosure include: PCT
Application Nos. WO 99/64002, WO 00/74679, WO 01/991752, WO
01/74844, WO 01/70708, WO 01/70337, WO 01/91752, WO 02/059095, WO
02/059107, WO 02/059108, WO 02/059117, wo 02/12166, WO 02111715, WO
02/12178, WO 02/15909, WO 02/068387, WO 02/068388, WO 02/067869, WO
03/007949, and WO 03/009847. Specific Mc4r agonists useful in the
present disclosure include CIR86036 (Chiron), ME-10142, and
ME-10145 (Melacure).
[0386] Monoamine reuptake inhibitors useful in the present
disclosure include: PCT Application Nos. WO 01/27068, and WO
01/62341. Specific monoamine reuptake inhibitors useful in the
present disclosure include, but are not limited to, sibutramine
(Meridia O/Reductil.RTM.) disclosed in U.S. Pat. Nos. 4,746,680,
4,806,570, and 5,436,272, and U.S. Patent Publication No.
2002/0006964.
[0387] Serotonin reuptake inhibitors, and releasers, useful in the
present disclosure include: dexfenfluramine, fluoxetine, and other
serotonin reuptake inhibitors, including, but not limited to, those
in U.S. Pat. No. 6,365,633, and PCT Patent Application Nos. WO
01/27060, and WO 01/162341.
[0388] 11.beta. HSD-1 inhibitor useful in the present disclosure
include, but are not limited to, BVT 3498, BVT 2733, and those
compounds disclosed in WO 01/90091, WO 01/90090, WO 01/90092.
Uncoupling Protein (UCP-1, UCP-2, and UCP-3) activators useful in
the present disclosure include: PCT Patent Application No. WO
99/00123. Specific uncoupling protein (UCP-1, UCP-2, and UCP-3)
activators useful in the present disclosure include, but are not
limited to, phytanic acid,
4-[(E)-2-(5,6,7,8-tetrahydro-5,5,8,8-tetramethyl-2-napthalenyl)-1-propeny-
l]benzoic acid (TTNPB), and retinoic acid.
[0389] .beta.3 adrenergic receptor (.beta.3) agonists useful in the
present disclosure include: U.S. Pat. Nos. 5,705,515 and 5,451,677
and PCT Patent Application Nos. WO 01/74782, and WO 02/32897.
Specific 3 agonists useful in the present disclosure include, but
are not limited to, AD9677/TAK677 (Dainippon/Takeda), CL-316,243,
SB 418790, BRL-37344, L-796568, BMS-196085, BRL-35135A, CGP12177A,
BTA-243, GW 427353, Trecadrine, Zeneca D7114, and SR 59119A.
[0390] Thyroid hormone p agonists useful in the present disclosure
include: PCT Application No. WO 02/15845 and Japanese Patent
Application No. JP 2000256190. Specific thyroid hormone 3 agonists
useful in the present disclosure include, but are not limited to,
KB-2611 (KaroBioBMS). Specific fatty acid synthase (PAS) inhibitors
useful in the present disclosure, include, but are not limited to,
Cerulenin and C75. Specific phosphodieterase (PDE) inhibitors
useful in the present disclosure, include, but are not limited to,
theophylline, pentoxifylline, zaprinast, sildenafil, arnrinone,
milrinone, cilostamide, rolipram, and cilomilast.
[0391] Lipase inhibitors useful in the present disclosure include,
but are not limited to, those disclosed in PCT Application No. WO
01/77094, and U.S. Pat. Nos. 4,598,089, 4,452,813, 5,512,565,
5,391,571, 5,602,151, 4,405,644, 4,189,438, and 4,242,453. Specific
lipase inhibitors useful in the present disclosure include, but are
not limited to, tetrahydrolipstatin (orlistat/Xenical.RTM.), Triton
WR1339, RHC80267, lipstatin, teasaponin, and diethylumbelliferyl
phosphate, FL-386, WAY-121898, Bay-N-3176, valilactone, esteracin,
ebelactone A, ebelactone B, and RHC 80267.
[0392] Examples of HMG-CoA reductase inhibitors include, but are
not limited to, lovastatin, simvastatin, pravastatin and
fluvastatin. Examples of HMG-CoA synthase inhibitors are the
beta-lactone derivatives disclosed in U.S. Pat. Nos. 4,806,564,
4,816,477, 4,847,271, and 4,751,237; the beta-lactam derivatives
disclosed in U.S. Pat. No. 4,983,597 and U.S. Ser. No. 07/540,992
filed Jun. 20, 1990; and the substituted oxacyclopropane analogues
disclosed in European Patent Publication EP 0 411 703. Examples of
squalene epoxidase inhibitors are disclosed in European Patent
Publication EP 0 318 860 and in Japanese Patent Publication J02
169-571A. Examples of LDL-receptor gene inducer molecules are
disclosed in U.S. Pat. No. 5,182,298 filed Mar. 18, 1991. Other
cholesterol lowering agents that may be administered include
niacin, probucol, fibric acids (i.e., clofibrate and gemfibrozil),
and LDL-receptor gene inducers.
[0393] Examples of PARP inhibitors include, but are not limited to,
iodonitocoumarin, 5-iodo-6-nitrocoumarin,
3,4-dihydro-5-methyl-isoquinolinone, 4-amino-1,8-naphthalimide,
3-methoxybenzamide, 8-hydroxy-2-methyl-3-hydro-quinazolin-4-one,
2-{3-[4-(4-fluorophenyl)-3,6-dihydro-1(2h)-pyridinyl]propyl}-8-methyl-4(3-
h)-quinazolinone,
5-fluoro-1-[4-(4-phenyl-3,6-dihydropyridin-1(butyl]quinazoline-2,4(1h,3h)-
-dione, 3-(4-chlorophenyl) quinoxaline-5-carboxamide,
2-(3'-methoxyphenyl)benzimidazole-4-carboxam, benzamide,
3-aminobenzamide, 3-aminophthalhydrazide, and
1,5-dihydroxyisoquinoline.
[0394] The above-mentioned compounds, which can be used in
combination with a compound of Formula (I) or (II), or a
pharmaceutically acceptable salt thereof, can be prepared and
administered as described in the art such as in the documents cited
above.
[0395] The above compounds are only illustrative of the ACMSD
inhibitors, anti-diabetic agents, anti-obesity agents, cholesterol
lower agent, compounds that boost NAD levels, compounds that
inhibit NAD.sup.+ consumption that can be used in the compositions
of the present disclosure. As this listing of compounds is not
meant to be comprehensive, the methods of the present disclosure
may employ any anti-obesity agent and any anti-diabetic agent, and
are not limited to any particular structural class of
compounds.
[0396] As used herein, "combination therapy" includes the
administration of a compound of the present disclosure, or a
pharmaceutically acceptable salt, prodrug, metabolite, polymorph or
solvate thereof, and at least a second agent as part of a specific
treatment regimen intended to provide the beneficial effect from
the co-action of these therapeutic agents. The beneficial effect of
the combination includes, but is not limited to, a cooperative,
e.g., synergistic, effect and/or a pharmacokinetic or
pharmacodynamic co-action, or any combination thereof, resulting
from the combination of therapeutic agents. Administration of these
therapeutic agents in combination typically is carried out over a
defined time period (usually minutes, hours, days or weeks
depending upon the combination selected). "Combination therapy" may
be, but generally is not, intended to encompass the administration
of two or more of these therapeutic agents as part of separate
monotherapy regimens that incidentally and arbitrarily result in
the combinations of the present disclosure.
[0397] "Combination therapy" is intended to embrace administration
of these therapeutic agents in a sequential manner, wherein each
therapeutic agent is administered at a different time and in any
order, or in alternation and in any order, as well as
administration of these therapeutic agents, or at least two of the
therapeutic agents, in a substantially simultaneous manner.
Substantially simultaneous administration can be accomplished, for
example, by administering to the subject a single capsule having a
fixed ratio of each therapeutic agent or in multiple, single
capsules for each of the therapeutic agents. Sequential or
substantially simultaneous administration of each therapeutic agent
can be effected by any appropriate route including, but not limited
to, oral routes, intravenous routes, intramuscular routes, and
direct absorption through mucous membrane tissues. The therapeutic
agents can be administered by the same route or by different
routes. For example, a first therapeutic agent of the combination
selected may be administered by intravenous injection while the
other therapeutic agents of the combination may be administered
orally. Alternatively, for example, all therapeutic agents may be
administered orally or all therapeutic agents may be administered
by intravenous injection. The sequence in which the therapeutic
agents are administered is not narrowly critical.
Biological Assays and Animals Studies
Method of Screening ACMSD1 Inhibition
[0398] The activity of compounds as inhibitors of ACMSD1 is
determined in a spectrophotometrical in vitro assay. The pre-assay
mixture is incubated and a compound of Formula (I) or (II), or a
pharmaceutically acceptable salt thereof, and ACMSD1 solution is
then added. The effect of ACMS concentration on the enzyme activity
is investigated by varying 3-hydroxyanthranilic acid (30H-HA)
concentration in the pre-assay mixture. Kinetic parameters are
calculated from the initial velocity data using a Lineweaver-Burk
plot.
Cellular Assay Methods
[0399] The mouse hepatocytes cell lines are grown and plated. The
cells are maintained in culture at 37.degree. C. and once the cells
are attached, different concentrations of a compound of Formula (I)
or (II), or a pharmaceutically acceptable salt thereof, or DMSO are
added. Primary hepatocytes are harvested about 24 hrs later.
Determination of ACMSD-1 Modulation in HEK293T Cells.
[0400] HEK293T cells are seeded and transfected to transiently
express ACMSD. The cells are then stimulated with different
concentrations of a compound of Formula (I) or (II), and then lysed
to measure the ACMSD activity in a spectrophotometrical in vitro
assay. The amount of the whole protein content in cell lysates is
detected by Bradford analysis and used to get the specificity
activity of the enzyme normalized in all samples.
Determination of NAD.sup.+ Content in Human Primary Hepatocytes
[0401] Primary hepatocytes are treated with different
concentrations of a compound of Formula (I) or (II), or a
pharmaceutically acceptable salt thereof, or MEHP (control) after
seeding. The compound is replaced every 24 hours, and then cells
are directly harvested and lysed to detect NAD.sup.+ content
through LC MS/MS (liquid chromatography mass spectrometry/mass
spectroscopy).
Modulation of SOD2 Activity in AML12 Cells and Murine Primary
Hepatocytes
[0402] Primary hepatocytes or AML-12 cells are lysed and total
protein concentration is determined using the Bradford assay. SOD2
activity is determined at indicated times after treatment with a
compound of Formula (I) or (II), or a pharmaceutically acceptable
salt thereof, using a SOD Assay Kit. Absorbance is determined and
results are expressed in U/ml/mg of protein according to the
standard curve and measured protein concentration.
Determination of NAD.sup.+ Content in Murine Primary
Hepatocytes
[0403] NAD.sup.+ is extracted using acidic extraction method and
samples are collected and homogenized. After insoluble protein
parts are pelleted, the samples are separated by high-performance
liquid chromatography (HPLC) and analyzed by mass-spectrometry. The
proteins in the pellet are quantified by Bradford assay and are
used for normalization.
RNA Preparation and RT-qPCR Analysis of ACMSD and SIRT1-Regulated
Genes in Cells
[0404] Cells (AML-12, Hepa-1.6, HEK-293, primary human and murine
hepatocytes) are treated with different concentrations of a
compound of Formula (I) or (II), or a pharmaceutically acceptable
salt thereof and the gene expression of ACMSD, Pgc1a, Sod1, and
Sod2 (MnSOD) is determined using RT-qPCR. Total RNA is extracted
from cells and the extracted RNA is treated with DNase and used for
reverse transcription (RT).
Modulation of Caspase 3/7 Activity in MDCK Cells
[0405] MDCK cells are cultured in base medium to a final
concentration of 10%. Cells are plated into 96 wells and 24 hours
after cell plating the medium is changed with fresh medium
supplemented with 1% FBS. Cisplatin is then used to induce cell
injury. Different concentrations of Formula (I) or (II), or a
pharmaceutically acceptable salt thereof (in DMSO) are added in
combination with cisplatin or prior to adding cisplatin. Caspase
3/7 activity (Promega) is determined according to standard
procedures using a luminescent signal readout on a plate reader.
Each experiment/condition is performed in triplicate. Caspase
activity is analyzed as percentage effect normalized to the
cisplatin alone and vehicle treated cells.
Cytotoxicity and hERG Screening
[0406] HePG2 and AML-12 cells are seeded and a dose-response of the
compound is performed at various concentrations. Cells are
stimulated and the supernatant is used to perform LDH release as a
measure of necrosis while the cells are lysed to detect ATP levels
for determining cell viability.
[0407] The Predictor hERG assay kit is stably transfected with hERG
potassium channel and a high-affinity red fluorescent hERG channel
ligand and is used for the determination of hERG channel affinity
binding of compounds of Formula (I) or (II), or a pharmaceutically
acceptable salt thereof. Compounds that bind to the hERG channel
protein (competitors) are identified by their ability to displace
the tracer which results in a lower fluorescence polarization.
C. elegans Experiments--ACMSD1 Silencing, Lifespan Assays, Mobility
Assessment and GFP Quantification
[0408] ACMSD1 silencing: Bacterial feeding RNAi experiments to
determine the effects of downregulation or silencing of acmsd-1 on
gene expression and survival are carried out in the nematode
Caenorhabditis elegans (C. elegans). The clones used for the
bacterial feeding experiments are acmsd-1, SIR-2.1 and DAF-16.
Total RNA is extracted from cells and the extracted RNA is treated
with DNase, and used for reverse transcription (RT).
[0409] Worms are grown on NGM agar plates additionally containing
Carbenicillin and IPTG and seeded with bacterial cultures. After
RNAi treatment, worms are transferred to plates containing paraquat
and seeded with RNAi bacteria. Control animals are grown on RNAi
bacteria containing an empty vector (control) and then transferred
to plates containing paraquat and seeded with RNAi bacteria.
Quantification of gene expression of sod-3 at mRNA levels and
protein levels using RT-qPCR and survival analyses are performed.
The movement of worms is recorded at days 1, 3, and 5 of
adulthood.
Anti-Diabetic Effects Studies in C57BL 6J and KK-Ay Mice
[0410] Mice are fed with regular chow or a high fat diet (HFD). A
compound of Formula (I), or a pharmaceutically acceptable salt
thereof, is dosed daily and blood and tissues are harvested for RNA
isolation, lipid measurements and histology. Oxygen consumption is
measured and histological analysis and transmission electron
microscopy are performed. An oral glucose tolerance test and an
intraperitoneal insulin tolerance test are also performed to
quantify glucose and to measure plasma insulin concentrations.
Anti-Diabetic and Anti-Obesity Studies in db/db Mice with LepR
Mutation
[0411] Animals are fed a high-fat diet (HFD). For subchronic
intervention, the animals are treated once/day with a compound of
Formula (I) or (II), or a pharmaceutically acceptable salt thereof,
for 14 days. Blood samples are collected and glucose concentrations
of each blood sample are determined. For acute intervention,
initial blood samples are collected and then compounds of Formula
(I) or (II), or a pharmaceutically acceptable salt thereof, are
administered. Diet-access is then restricted, and a second blood
sample is collected. The mice are subjected to an oral glucose
tolerance test and blood glucose concentrations are determined.
[0412] For the euglycemic-hyperinsulinemic clamps assay, the
animals receive a primed-continuous [3-.sup.3H]glucose infusion and
a blood sample is then collected to determine plasma insulin,
glucose and [3-.sup.3H]glucose concentrations and to calculate
basal endogenous glucose appearance rates. The mice then receive
vehicle or a compound of Formula (I) or (II), or a pharmaceutically
acceptable salt thereof, via gavage. Subsequently, the animals
receive a [3-.sup.3H]glucose infusion containing insulin causing a
moderate net-increase in plasma insulin concentrations. Blood
glucose concentrations are measured and target glycemia is
established by adjusting the rate of glucose infusion.
2-deoxy-D-[1-.sup.14C] glucose is then given intravenously and
blood samples are collected. The mice are then sacrificed.
Gastrocnemius muscle and epididymal adipose tissue are collected
and plasma [.sup.3H]- and [.sup.14C]-radioactivity is determined in
deproteinized plasma.
[0413] Body weights are assessed and brown adipose tissue (BAT) and
gonadal white adipose tissue (WAT) are dissected and weighed.
Volume oxygen (VO.sub.2) and volume carbon dioxide production
(VCO.sub.2) are measured and are reported as average VO.sub.2 per
hour normalized to body weight (mL/h/kg). Activity counts by
infrared beam interruptions and food intake are simultaneously
measured.
Non-Alcoholic Fatty Liver Disease (NAFLD) and Non-Alcoholic
Steatohepatitis (NASH) Studies in Male C57BL 6J Mice
[0414] Mice are fed a `Western` HF-HSD (high fat-high sucrose diet)
or normal chow diet (NCD) as control. The animals are then treated
with a compound of Formula (I) or (II), or a pharmaceutically
acceptable salt thereof, for 4, 12 or 20 weeks, and then
sacrificed. Body weight and food intake are monitored weekly and
total fat mass is analysed. An intraperitoneal glucose tolerance
test (IPGTT) is also performed and tail vein glucose levels are
measured after glucose administration. Insulin resistance is
calculated using the Homeostasis Model of Insulin Resistance. The
mice are then sacrificed by blood sampling via cardiac puncture.
Plasma is obtained and tissues were collected together with the
plasma for further biochemical and molecular analyses or for
histological analysis.
Non-Alcoholic Fatty Liver Disease (NAFLD) and Non-Alcoholic
Steatohepatitis (NASH) Studies in in Methionine and Choline
Deficient Mice
[0415] Mice weighing 25 g are either fed a methionine- and
choline-deficient diet (MCD to induce NASH) or chow diet (as a
control). Animal experiments and evaluation of NAFLD and NASH are
conducted as described above in for C57BL/6J mice fed the high fat
and high sucrose diet.
Atherosclerosis Studies in High Cholesterol Fed LDL-R Knockout
Mice
[0416] LDL-R knockout (KO) mice are sacrificed about 12 weeks after
the initiation of the atherogenic diet, after which the heart and
aorta are perfused with PBS and subsequently fixed. Atherosclerosis
and biochemistry parameters are measured with the appropriate
commercially available kits. For the in vivo lipopolysaccharide
(LPS) study, mice are intraperitoneally injected with LPS, and
blood is taken from the tail vein. TNF.alpha. levels are quantified
with a Mouse TNF.alpha. ELISA assay. Blood cell counts are
determined.
Inherited Mitochondrial Disease Studies in Sco2.sup.KO/KI Mice
[0417] Compounds of Formula (I) or (II), or a pharmaceutically
acceptable salt thereof, are dissolved in water and added to a
standard powder diet at the appropriate concentration. The diet
supply is changed every three days and administered ad libitum for
one month. Tissues are collected for histological analysis. For the
muscle quadriceps samples, the spectrophotometric activity of cI,
cII, cIII, and cIV, as well as CS, is measured. NAD.sup.+ is
extracted from tissues using acidic and alkaline extraction
methods, respectively, and analysed with mass spectrometry.
Inherited Mitochondrial Disease Studies in Deletor Mice
[0418] Deletor and WT male mice are administered either chow diet
(CD) or a compound of Formula (I) or (II), or a pharmaceutically
acceptable salt thereof, admixed with the CD. The mice are
regularly monitored for weight, food consumption, and physical
endurance and their exercise capability is measured. Oxygen
consumption and carbon dioxide production, as well as spontaneous
moving and feeding activities, are recorded. Tissue sections are
collected and prepared from the quadriceps, liver, and BAT. Frozen
sections from quadriceps are assayed for in situ histochemical COX
and succinate dehydrogenase (SDH) activities, crista content in
both BAT and muscle is determined from electron micrographs and
skeletal muscle samples are analysed for citrate synthase
activity.
Kidney Disease Studies
[0419] C57BL/6J WT mice are fed a standard commercial diet and
divided into four groups: control; cisplatin; a compound of Formula
(I) or (II), or a pharmaceutically acceptable salt thereof, alone.
The mice are sacrificed and tissue samples and serum are collected.
Serum creatinine and BUN levels are measured and the
proinflammatory cytokines TNF-.alpha., IL-1b, and IL-6 from serum
or homogenates from kidney tissue are quantified. Mouse kidneys are
collected and stained for analysis. Tubular damage is examined and
scored based on the percentage of cortical tubular necrosis.
Neutrophil infiltration is quantitatively assessed on stained
tissue by counting the number of neutrophils per high-power
field.
[0420] Alternatively, C57BL/6J WT mice are numbered and kept in
acclimatization for a period and then randomized into different
treatment groups based on their body weight. Different groups are
maintained on a specified diet for a period of time. Body weight
measurements are taken and food consumption is evaluated. Blood is
collected by retro-orbital puncture under mild anesthesia and used
for analysis of basal blood urea nitrogen levels (BUN).
[0421] Mice are anesthetized and placed on a surgical platform.
Both kidneys are exposed through incisions and renal pedicles are
occluded using vascular clamps. The clamp is then removed and the
surgical site is sutured. The sham-operated group is subjected to
similar surgical procedures, except that the occluding clamp is not
applied. Animals are monitored until recovery from anesthesia and
returned to their home cage. Animals are observed every day for
general clinical signs and symptoms and mortality.
[0422] One day prior to termination, animals are individually
housed in metabolic cages and urine is collected for estimation of
urea, creatinine, sodium and potassium. Blood is also collected by
retro orbital puncture under mild anesthesia and plasma is used for
analysis of blood urea nitrogen levels (BUN) and serum creatinine.
Animals are then euthanized and organs are collected. One kidney is
fixed and the other is flash frozen and used for the estimation of
lipid peroxidation, GSH, MPO and SOD levels.
Ischemia Reperfusion-Induced Acute Kidney Injury Studies
[0423] CD-1 (ICR) mice are treated with a compound of Formula (I)
or (II), or a pharmaceutically acceptable salt thereof, by oral
gavage once per day. CD-1 mice are divided into four groups: (1)
young mice with sham injury; (2) young mice with
ischemic/reperfusion (I/R) injury; (3) adult mice with sham injury;
and (4) adult mice with I/R injury.
[0424] An additional 27 adult mice are randomized into two groups:
mice receiving a compound of Formula (I) or (II), or a
pharmaceutically acceptable salt thereof, and mice receiving the
vehicle as a control. The serum creatinine level is measured and
BUN measurements are recorded. Renal tissue is then evaluated and
tubular injury is scored.
Cisplatin-Induced Acute Kidney Injury Studies
[0425] C57BL6 mice are treated with compounds of Formula (I) or
(II), or a pharmaceutically acceptable salt thereof by oral gavage
once per day. The animals are allowed to recover, and they are
sacrificed 48,72, and 96 hrs post cisplatin injection.
[0426] The serum creatinine level is measured as a primary
endpoint. Tubular injury is scored on a scale of 0-4 on the basis
of the percentage of tubules with necrosis, dilatation, or cell
swelling: 0, less than 5%; 1, 5-25%; 2, 25-50%; 3, 50-75%; and 4,
over 75%. All high-power fields (.times.400) in the cortex and
outer medulla are evaluated by a pathologist in a blinded
manner.
Effects on Sepsis-Induced Acute Kidney Injury
[0427] C57BL6 mice (12-15 weeks old). are treated with compounds of
Formula (I) or (II), or a pharmaceutically acceptable salt thereof,
via IP injection following Cecal ligation and puncture induced
Sepsis.
[0428] Blood and kidney tissues are collected at the time of
sacrificing for measurement of primary and secondary endpoints.
Primary endpoint (at 48 hrs):serum creatinine. Secondary endpoints
(at 48 hours) include: Macrophage phenotype marker (IF stain),
Plasma NGAL, Plasma and kidney markers of inflammation (IL-6,
IL-18, TNF), and Kidney Injury markers (KIM-1, NGAL, TIMP2 and
IGFBP7). Addition endpoints include; cell death (IF: Annexin V and
Propidium Iodide; Caspase 3/7), autophagy, biogenesis
(PGC-1.alpha., mitochondrial DNA), OXPHOS (Complex I, III, IV
activity), Sirt1 and Sirt3 expression, AMPK (Total, P-AMPK, P-ACC,
and HIF-1.alpha..
[0429] Histological analysis is performed with H&E and PAS
staining using standard protocols. Images are collected and
analyzed using a light microscope (IX71, Olympus, Tokyo, Japan)
with DP analyzer software (DP70-BSW, Tokyo, Japan). Tubular damage
in PAS-stained kidney sections is scored based on the percentage of
cortical tubular necrosis: 0=normal, 1=1-10, 2=11-25, 3=26-45,
4=46-75, and 5=76-100%. tubular injury score will be used to
evaluate protection against kidney damage.
Determination of the Effects on FoxO1 Phosphorylation Levels
[0430] AML-12 cells are treated with different concentrations of a
compound of Formula (I) or (II), or a pharmaceutically acceptable
salt thereof. Cells are then lysed, and analyzed by
SDS-PAGE/western blot. Blocking and antibody incubations are then
done and each protein present is detected with its specific
antibody.
Inhibitory Effect
[0431] The present disclosure also relates to a compound of Formula
(I) or (II), or a pharmaceutically acceptable salt thereof, as
defined herein, in a method for inhibiting the activity of ACMSD.
The method includes contacting a cell with a compound of Formula
(I) or (II), or a pharmaceutically acceptable salt thereof. In a
related embodiment, the method further provides that the compound
is present in an amount effective to produce a concentration
sufficient to selectively inhibit ACMSD in the cell.
[0432] Thus, preferably in an assay for ACMSD inhibition (i.e., an
ACMSD assay described herein, e.g., Biological Example 1, or an
ACMSD assays known in the literature), the preferred compounds of
Formula (I) or (II), or a pharmaceutically acceptable salt thereof,
are compounds capable of reducing or preferably inhibiting ACMSD
and increasing NAD.sup.+ levels and/or activating SIRTs and the
downstream targets of SIRTs, such as PGC-1.alpha., FoxO1 and/or
SOD. Preferably, said inhibition is determined as the IC.sub.50 of
said compound of Formula (I) or (II), or a pharmaceutically
acceptable salt thereof, with respect to said ACMSD inhibition
assay. Preferred compounds of Formula (I) or (II), or a
pharmaceutically acceptable salt thereof, have an IC.sub.50 at or
below 1 .mu.M, more preferably less than 300 nM, for example less
than 100 nM, such as less than 50 nM with respect to inhibition of
ACMSD.
EXEMPLARY EMBODIMENTS
[0433] Some embodiments of this disclosure are Embodiment I, as
follows:
[0434] Embodiment I-1. A compound represented by Formula (I):
##STR00165##
[0435] or a pharmaceutically acceptable salt or tautomer
thereof,
[0436] wherein:
[0437] X is H, S, SR.sup.2, NR.sup.2, NR.sup.2R.sup.2', O, OH,
OR.sup.h, F, Br, or Cl;
[0438] W is N or C; [0439] (i) when W is N, then: [0440] L is
--(C(R.sup.5).sub.2).sub.mCH.dbd.CH(C(R.sup.5).sub.2).sub.p--,
[0440] ##STR00166##
--(C(R.sup.5).sub.2).sub.mY.sup.1(C(R.sup.5).sub.2).sub.p--,
--(C(R.sup.5).sub.2).sub.mY.sup.1(C(R.sup.5).sub.2).sub.pcyclopropyl-,
--(C(R.sup.5).sub.2).sub.mY.sup.1CH.dbd.CH--,
--(C(R.sup.5).sub.2).sub.mNR.sup.3C.dbd.(O)(C(R.sup.5).sub.2).sub.p--,
--(C(R.sup.5).sub.2).sub.mphenyl(C(R.sup.5).sub.2).sub.p--,
--(C(R.sup.5).sub.2).sub.mpyridinyl(C(R.sup.5).sub.2).sub.p--, or
--(C(R.sup.5).sub.2).sub.mthiophenyl(C(R.sup.5).sub.2).sub.p--;
[0441] (ii) when W is C, then: [0442] L is
--(C(R.sup.5).sub.2).sub.mCH.dbd.CH(C(R.sup.5).sub.2).sub.p,
(C(R.sup.5).sub.2).sub.o,
--(C(R.sup.5).sub.2).sub.mY.sup.1(C(R.sup.5).sub.2).sub.p,
##STR00167##
[0442] --(C(R.sup.5).sub.2).sub.m Y.sup.1CH.dbd.CH--,
--(C(R.sup.5).sub.2).sub.mC.dbd.(O)(CH.sub.2).sub.p,
--(C(R.sup.5).sub.2).sub.mC.dbd.(O)O(C(R.sup.5).sub.2).sub.p--,
--(C(R.sup.5).sub.2).sub.mC.dbd.(O)NR.sup.3(C(R.sup.5).sub.2).sub.p--,
--(C(R.sup.5).sub.2).sub.mNR.sup.3C.dbd.(O)(C(R.sup.5).sub.2).sub.p--,
--(C(R.sup.5).sub.2).sub.mphenyl(C(R.sup.5).sub.2).sub.p--,
--(C(R.sup.5).sub.2).sub.mpyridinyl(C(R.sup.5).sub.2).sub.p--, or
--(C(R.sup.5).sub.2).sub.mthiophenyl(C(R.sup.5).sub.2).sub.p--;
[0443] Y.sup.1 is O, NR.sup.4, or S(O).sub.q;
[0444] each Y.sup.2 is independently O, NH or S;
[0445] R.sup.1 is absent or C.sub.6-C.sub.10 arylene or
heteroarylene, wherein the heteroarylene comprises one or two 5- to
7-membered rings and 1-4 heteroatoms selected from N, O and S, and
wherein the C.sub.6-C.sub.10 arylene or heteroarylene are
optionally substituted with one to two R.sup.e;
[0446] R.sup.2 is H or C.sub.1-C.sub.4 alkyl;
[0447] R.sup.2' is H, C.sub.1-C.sub.4 alkyl, or C.sub.3-C.sub.7
cycloalkyl; or
[0448] R.sup.2 and R.sup.2' together with the nitrogen atom to
which they are attached form a 3- to 7-membered heterocycloalkyl
ring comprising 1-3 additional heteroatoms selected from N, O and
S;
[0449] R.sup.3 is H or C.sub.1-C.sub.4 alkyl;
[0450] R.sup.4 is H or C.sub.1-C.sub.4 alkyl;
[0451] each R.sup.5 is independently at each occurrence H or
C.sub.1-C.sub.4 alkyl;
[0452] each R.sup.6 is independently at each occurrence H or
C.sub.1-C.sub.4 alkyl;
[0453] R.sup.7 is H, A, B, or C;
[0454] A is --(C(R.sup.6).sub.2).sub.rCO.sub.2R.sup.x,
--Y.sup.2(C(R.sup.6).sub.2).sub.rCO.sub.2R.sup.x,
--(CH.sub.2).sub.rtetrazole, --(CH.sub.2).sub.roxadiazolone,
--(CH.sub.2).sub.rtetrazolone, --(CH.sub.2).sub.rthiadiazolol,
--(CH.sub.2).sub.r isoxazol-3-ol,
--(CH.sub.2).sub.rP(O)(OH)OR.sup.x, --(CH.sub.2).sub.rS(O).sub.2OH,
--(CH.sub.2).sub.rC(O)NHCN, or
--(CH.sub.2).sub.rC(O)NHS(O).sub.2alkyl, wherein
--(CH.sub.2).sub.rtetrazole, --(CH.sub.2).sub.roxadiazolone,
--(CH.sub.2).sub.rtetrazolone, --(CH.sub.2).sub.rthiadiazolol,
--(CH.sub.2).sub.r isoxazol-3-ol are optionally substituted with
C.sub.1-C.sub.6 alkyl,
[0455] B is --(C(R.sup.6).sub.2).sub.rS(O).sub.2OC.sub.1-C.sub.4
alkyl, --O(C(R.sup.6).sub.2).sub.rS(O).sub.2OC.sub.1-C.sub.4 alkyl,
--Y.sup.2(C(R.sup.6).sub.2).sub.rC(O)NR.sup.gR.sup.g',
--Y.sup.2(C(R.sup.6).sub.2).sub.rS(O).sub.2NR.sup.gR.sup.g',
--(CH.sub.2).sub.rC(O)NR.sup.gR.sup.g',
--(CH.sub.2).sub.rS(O).sub.2NR.sup.gR.sup.g',
--(CH.sub.2).sub.rC(O)NHS(O).sub.2NR.sup.gR.sup.g',
--(C(R.sup.6).sub.2).sub.rCO.sub.2R.sup.i,
--(C(R.sup.6).sub.2).sub.rNH.sub.2CO.sub.2R.sup.x,
--(C(R.sup.6).sub.2).sub.rP(O)(OR.sup.x).sub.2,
--O(C(R.sup.6).sub.2).sub.rP(O)(OR.sup.x).sub.2,
--(C(R.sup.6).sub.2).sub.rS(O).sub.2OH,
--O(C(R.sup.6).sub.2).sub.rS(O).sub.2OH,
--(C(R.sup.6).sub.2).sub.rP(O).sub.2OR.sup.x, or
--O(C(R.sup.6).sub.2).sub.rP(O).sub.2OR.sup.x,
[0456] C is --(CH.sub.2).sub.rCN, --(CH.sub.2).sub.sOH, halogen,
--(C(R.sup.6).sub.2).sub.rC.sub.6-C.sub.10 aryl,
--(C(R.sup.6).sub.2).sub.rS--C.sub.6-C.sub.10 aryl,
--(C(R.sup.6).sub.2).sub.rheteroaryl,
--O(C(R.sup.6).sub.2).sub.rheteroaryl,
--O(C(R.sup.6).sub.2).sub.rheterocycloalkyl,
--O(C(R.sup.6).sub.2).sub.rOH, --OR.sup.y,
--(C(R.sup.6).sub.2).sub.rC(O)NHCN, --CH.dbd.CHCO.sub.2R.sup.x, or
--(C(R.sup.6).sub.2).sub.rC(O)NHS(O).sub.2C.sub.1-C.sub.4 alkyl,
wherein the aryl and heteroaryl are substituted with one to three
substituents each independently selected from C.sub.1-C.sub.6
alkyl, C.sub.1-C.sub.6 haloalkyl, halogen, and OH, and wherein the
heterocycloalkyl is substituted with one to two .dbd.O or
.dbd.S;
[0457] R.sup.c is H, C.sub.1-C.sub.6 alkyl, C.sub.1-C.sub.6
haloalkyl, halogen, --CN, --OR.sup.x, or --CO.sub.2R.sup.x;
[0458] R.sup.d is methyl, CF.sub.3, CR.sup.fF.sub.2,
--(C(R.sup.6).sub.2).sub.tC.sub.6-C.sub.10 aryl,
--(C(R.sup.6).sub.2).sub.t-5- or 6-membered heteroaryl,
--(C(R.sup.6).sub.2).sub.t-5- or 6-membered cycloalkyl, optionally
substituted C.sub.6-C.sub.10 aryl, optionally substituted 5- or
6-membered heteroaryl, or optionally substituted 5- or 6-membered
cycloalkyl;
[0459] each R.sup.e is independently at each occurrence
C.sub.1-C.sub.6 alkyl, C.sub.2-C.sub.6 alkenyl, C.sub.2-C.sub.6
alkynyl, halogen, C.sub.1-C.sub.6 haloalkyl, --NHR.sup.z, --OH, or
--CN;
[0460] R.sup.f is absent, H, or methyl;
[0461] R.sup.g is H, C.sub.1-C.sub.6 alkyl, OH,
--S(O).sub.2(C.sub.1-C.sub.6 alkyl), or S(O).sub.2N(C.sub.1-C.sub.6
alkyl).sub.2;
[0462] R.sup.g' is H, C.sub.1-C.sub.6 alkyl, C.sub.3-C.sub.7
cycloalkyl, 4- to 7-membered heterocycloalkyl ring comprising 1-3
heteroatoms selected from N, O and S, C.sub.6-C.sub.10 aryl, or 5-
to 7-membered heteroaryl comprising 1-3 heteroatoms selected from
N, O and S, wherein the alkyl is optionally substituted with one or
more substituents independently selected from halogen and --OH, and
wherein the cycloalkyl, heterocycloalkyl, aryl and heteroaryl are
optionally substituted with one or more substituents independently
selected from C.sub.1-C.sub.6 alkyl, halogen, and --OH;
[0463] R.sup.h is H, C.sub.1-C.sub.4 alkyl, or 3- to 7-membered
heterocycloalkyl ring comprising 1-3 heteroatoms selected from N, O
and S, wherein the alkyl is optionally substituted with one or more
substituents each independently selected from NH.sub.2,
C.sub.1-C.sub.4 alkylamino, C.sub.1-C.sub.4 dialkylamino, and
C(O)NH.sub.2; and wherein the heterocycloalkyl is optionally
substituted with one or more substituents each independently
selected from C.sub.1-C.sub.6 alkyl and C.sub.1-C.sub.6
haloalkyl;
[0464] R.sup.i is (i) --(CH.sub.2).sub.sOC(O)C.sub.1-C.sub.6 alkyl,
wherein the alkyl is substituted with one or more NH.sub.2; (ii)
(CH.sub.2CH.sub.2O).sub.nCH.sub.2CH.sub.2OH; or (iii)
C.sub.1-C.sub.6 alkyl substituted with one or more substituents
each independently selected from OH and 4- to 7-membered
heterocycloalkyl comprising 1 to 3 heteroatoms selected from O, N,
or S;
[0465] R.sup.j is absent, H, C.sub.1-C.sub.6 alkyl, or --CN;
[0466] each R.sup.x is independently at each occurrence H,
C.sub.1-C.sub.6 alkyl, or C.sub.6-C.sub.10 aryl;
[0467] each R.sup.y and R.sup.z is independently H, C.sub.1-C.sub.6
alkyl, or C.sub.1-C.sub.6 haloalkyl;
[0468] each m, p, q, r, and t is independently 0, 1 or 2;
[0469] n is 0, 1, 2, or 3;
[0470] s is 1 or 2;
[0471] o is 0, 1, 2, 3, or 4; and
[0472] represents a single bond or a double bond; and
[0473] provided that
[0474] when X is O; R.sup.f is H; W is C; R.sup.j is --CN; L is
--SCH.sub.2--; R.sup.1 is phenylene or pyridine; then R.sup.7 is
not --COOH;
[0475] when X is O; R.sup.f is H; W is C; R.sup.j is --CN; L is
--SCH.sub.2--; R.sup.1 is phenylene or pyridine; and R.sup.7 is
tetrazole; then R.sup.c is not H;
[0476] when X is O; R.sup.f is H; W is C; R.sup.j is --CN; L is
--S--C(R.sup.5).sub.2 or --SCH.sub.2CH.sub.2--; R.sup.1 is absent;
then R.sup.7 is not COOH or tetrazole;
[0477] when X is O, R.sup.f is H; W is N; R.sup.i is absent;
R.sup.d is methyl, optionally substituted 5- to 10-membered aryl,
optionally substituted 5- or 6-membered heteroaryl, or optionally
substituted 5- or 6-membered cycloalkyl; L is --SCH.sub.2-- or
--OCH.sub.2--; and R.sup.1 is phenylene; then R.sup.7 is not
--COOH, --CH.sub.2COOH,
##STR00168##
and
[0478] when X is O, R.sup.f is H, W is N, R.sup.j is absent, L is
--NHCH.sub.2--, --CH.sub.2NH--, or --NH--C(O)--, and R.sup.1 is
phenylene, then R.sup.d is not phenyl.
[0479] Embodiment I-2. The compound of Embodiment I-1, wherein X is
O, OH, OR.sup.h, F, Br, or Cl.
[0480] Embodiment I-3. The compound of Embodiment I-1, wherein X is
H, S, SR.sup.2, NR.sup.2, or NR.sup.2R.sup.2'.
[0481] Embodiment I-4. The compound of any one of Embodiments I-1
to I-3, wherein R.sup.f is absent.
[0482] Embodiment I-5. The compound of any one of Embodiments I-1
to I-3, wherein R.sup.f is H or methyl.
[0483] Embodiment I-6. The compound of any one of Embodiments I-1
to I-5, wherein W is N.
[0484] Embodiment I-7. The compound of Embodiment I-6, wherein
R.sup.i is absent.
[0485] Embodiment I-8. The compound of any one Embodiments I-1 to
I-5, wherein W is C.
[0486] Embodiment I-9. The compound of Embodiment I-8, wherein
R.sup.i is H, C.sub.1-C.sub.6 alkyl, or --CN.
[0487] Embodiment I-10. The compound of Embodiment I-8 or I-9,
wherein R.sup.i is --CN.
[0488] Embodiment I-11. The compound of any one of Embodiments I-1
to I-10, wherein R.sup.c is C.sub.1-C.sub.6 alkyl, --CN, or
halogen.
[0489] Embodiment I-12. The compound of any one of Embodiments I-1
to I-11, wherein R.sup.c is --CN or halogen.
[0490] Embodiment I-13. The compound of any one of Embodiments I-1
to I-12, wherein R.sup.c is --CN.
[0491] Embodiment I-14. The compound of any one of Embodiments I-1
to I-13, wherein R.sup.d is methyl.
[0492] Embodiment I-15. The compound of any one of Embodiments I-1
to I-13, wherein R.sup.d is optionally substituted 5- to
10-membered aryl.
[0493] Embodiment I-16. The compound of any one of Embodiments I-1
to I-13, wherein R.sup.d is optionally substituted 5- or 6-membered
heteroaryl.
[0494] Embodiment I-17. The compound of any one of Embodiments I-1
to I-13, wherein R.sup.d is optionally substituted 5- or 6-membered
cycloalkyl.
[0495] Embodiment I-18. The compound of any one of Embodiments I-1
to I-13, wherein R.sup.d is methyl, cyclohexyl, pyridinyl,
thiazolyl, phenyl, or thienyl.
[0496] 19. The compound of any one of Embodiments I-1 to I-13,
wherein R.sup.d is methyl, cyclohexyl, pyridinyl, thiazolyl,
thienyl, or optionally substituted phenyl.
[0497] Embodiment I-20. The compound of any one of Embodiments I-1
to I-13, wherein R.sup.d is methyl.
[0498] Embodiment I-21. The compound of any one of Embodiments I-1
to I-13, wherein R.sup.d is --CF.sub.3.
[0499] Embodiment I-22. The compound of any one of Embodiments I-1
to I-13, wherein R.sup.d is CR.sup.fF.sub.2.
[0500] Embodiment I-23. The compound of any one of Embodiments I-1
to I-13, wherein R.sup.d is
--(C(R.sup.6).sub.2).sub.rC.sub.6-C.sub.10 aryl,
--(C(R.sup.6).sub.2).sub.r-5- or 6-membered heteroaryl,
--(C(R.sup.6).sub.2).sub.r-5- or 6-membered cycloalkyl.
[0501] Embodiment I-24. The compound of any one of Embodiments I-1
to I-13, wherein R.sup.d is
--(C(R.sup.6).sub.2).sub.rC.sub.6-C.sub.10 aryl.
[0502] Embodiment I-25. The compound of any one of Embodiments I-1
to I-24, wherein L is
--(C(R.sup.5).sub.2).sub.mY.sup.1(C(R.sup.5).sub.2).sub.p--.
[0503] Embodiment I-26. The compound of Embodiment I-25, wherein
Y.sup.1 is S.
[0504] Embodiment I-27. The compound of any one of Embodiments I-1
to I-24, wherein L is
--(C(R.sup.5).sub.2).sub.mNR.sup.3C.dbd.(O)(C(R.sup.5).sub.2).sub.p--
or
--(C(R.sup.5).sub.2).sub.mY.sup.1(C(R.sup.5).sub.2).sub.pcyclopropyl-.
[0505] Embodiment I-28. The compound of any one of Embodiments I-1
to I-27, wherein R.sup.1 is C.sub.6-C.sub.10 arylene.
[0506] Embodiment I-29. The compound of any one of Embodiments I-1
to I-27, wherein R.sup.1 is heteroarylene.
[0507] Embodiment I-30. The compound of any one of Embodiments I-1
to I-27, wherein R.sup.1 is absent.
[0508] Embodiment I-31. The compound of any one of Embodiments I-1
to I-30, wherein R.sup.7 is A.
[0509] Embodiment I-32. The compound of Embodiment I-31, wherein A
is --(C(R.sup.6).sub.2).sub.rCO.sub.2R.sup.x or
--(CH.sub.2).sub.rtetrazole, wherein the
--(CH.sub.2).sub.rtetrazole is optionally substituted with
C.sub.1-C.sub.6 alkyl.
[0510] Embodiment I-33. The compound of any one of Embodiments I-1
to I-30, wherein R.sup.7 is B.
[0511] Embodiment I-34. The compound of Embodiment I-31, wherein B
is --(CH.sub.2).sub.rC(O)NR.sup.gR.sup.g', or
--(CH.sub.2).sub.rS(O).sub.2NR.sup.gR.sup.g',
[0512] Embodiment I-35. The compound of any one of Embodiments I-1
to I-30, wherein R.sup.7 is C.
[0513] Embodiment I-36. The compound of Embodiment I-31, wherein C
is --(CH.sub.2).sub.rCN, --(CH.sub.2).sub.sOH, or
--(C(R.sup.6).sub.2).sub.rC.sub.6-C.sub.10 aryl, wherein the aryl
is substituted with one to three substituents each independently
selected from C.sub.1-C.sub.6 alkyl, C.sub.1-C.sub.6 haloalkyl,
halogen, and OH.
[0514] Embodiment I-37. A compound, or a pharmaceutically
acceptable salt or tautomer thereof, selected from the group
consisting of
TABLE-US-00004 Cpd No. Structure I-1 ##STR00169## I-2 ##STR00170##
I-3 ##STR00171## I-4 ##STR00172## I-5 ##STR00173## I-6 ##STR00174##
I-7 ##STR00175## I-8 ##STR00176## I-9 ##STR00177## I-10
##STR00178## I-11 ##STR00179## I-12 ##STR00180## I-13 ##STR00181##
I-14 ##STR00182## I-15 ##STR00183## I-16 ##STR00184## I-17
##STR00185## I-18 ##STR00186## I-19 ##STR00187## I-20 ##STR00188##
I-21 ##STR00189## I-22 ##STR00190## I-23 ##STR00191## I-24
##STR00192## I-25 ##STR00193## I-26 ##STR00194## I-27 ##STR00195##
I-28 ##STR00196## I-29 ##STR00197## I-30 ##STR00198## I-31
##STR00199## I-32 ##STR00200## I-33 ##STR00201## I-34 ##STR00202##
I-35 ##STR00203## I-36 ##STR00204##
[0515] Embodiment I-38. A compound, or a pharmaceutically
acceptable salt or tautomer thereof, selected from the group
consisting of
##STR00205## ##STR00206## ##STR00207## ##STR00208## ##STR00209##
##STR00210## ##STR00211## ##STR00212## ##STR00213##
[0516] Embodiment I-39. A pharmaceutical composition comprising a
compound of any one of Embodiments I-1 to I-38, or a
pharmaceutically acceptable salt thereof, and at least one of a
pharmaceutically acceptable carrier, diluent, or excipient.
[0517] Embodiment I-40. The pharmaceutical composition according to
Embodiment I-39, which comprises one or more further therapeutic
agents.
[0518] Embodiment I-41. A method of treating, preventing, or
reducing the risk of a disease or disorder inhibited by
.alpha.-amino-.beta.-carboxymuconate-.epsilon.-semialdehyde
decarboxylase (ACMSD) comprising administering to the subject
suffering from or susceptible to developing the disease or disorder
a therapeutically effective amount of one or more compounds of any
one of Embodiments I-1 to I-38, or a pharmaceutically acceptable
salt thereof.
[0519] Embodiment I-42. A method of treating, preventing, or
reducing the risk of a disease or disorder associated with reduced
nicotinamide adenine dinucleotide (NAD.sup.+) levels comprising
administering to the subject suffering from or susceptible to
developing a disease or disorder associated with reduced NAD.sup.+
levels a therapeutically effective amount of one or more compounds
of any one of Embodiments I-1 to I-38, or a pharmaceutically
acceptable salt thereof.
[0520] Embodiment I-43. The method of any one of Embodiments I-41
to I-42, wherein the disease is chronic liver disease selected from
primary biliary cirrhosis (PBC), cerebrotendinous xanthomatosis
(CTX), primary sclerosing cholangitis (PSC), drug induced
cholestasis, intrahepatic cholestasis of pregnancy, parenteral
nutrition associated cholestasis (PNAC), bacterial overgrowth or
sepsis associated cholestasis, autoimmune hepatitis, chronic viral
hepatitis, alcoholic liver disease, nonalcoholic fatty liver
disease (NAFLD), nonalcoholic steatohepatitis (NASH), liver
transplant associated graft versus host disease, living donor
transplant liver regeneration, congenital hepatic fibrosis,
choledocholithiasis, granulomatous liver disease, intra- or
extrahepatic malignancy, Sjogren's syndrome, Sarcoidosis, Wilson's
disease, Gaucher's disease, hemochromatosis, and alpha
1-antitrypsin deficiency.
[0521] Embodiment I-44. A method of treating a disorder associated
with mitochondrial dysfunction comprising administering to the
subject suffering from or susceptible to developing a metabolic
disorder a therapeutically effective amount of one or more
compounds of any one of Embodiments I-1 to I-38, or a
pharmaceutically acceptable salt thereof, that increases
intracellular nicotinamide adenine dinucleotide (NAD.sup.+).
[0522] Embodiment I-45. The method of Embodiment I-44, wherein said
disorder associated with mitochondrial dysfunction is an inherited
mitochondrial disease, a common metabolic disorder, a
neurodegenerative disease, an aging related disorder, a kidney
disorder, or a chronic inflammatory disease.
[0523] Embodiment I-46. The method of Embodiment I-45, wherein the
common metabolic disorder is obesity or type II diabetes.
[0524] Embodiment I-47. A method of promoting oxidative metabolism
comprising administering to the subject suffering from or
susceptible to developing a metabolic disorder a therapeutically
effective amount of one or more compounds of any one of Embodiments
I-1 to I-38, or a pharmaceutically acceptable salt thereof, that
increases intracellular nicotinamide adenine dinucleotide
(NAD.sup.+).
[0525] Embodiment I-48. A compound of any of Embodiments I-1 to
I-38, or a pharmaceutically acceptable salt thereof, for use as a
medicament.
[0526] Embodiment I-49. A compound of any of Embodiments I-1 to
I-38, or a pharmaceutically acceptable salt thereof, for use in
treating, preventing, or reducing the risk of a disease or disorder
associated with reduced nicotinamide adenine dinucleotide
(NAD.sup.+) levels.
[0527] Embodiment I-50. A compound of any of Embodiments I-1 to
I-38, or a pharmaceutically acceptable salt thereof, for use in for
treating, preventing, or reducing the risk of a disorder associated
with mitochondrial dysfunction.
[0528] Embodiment I-51. A compound of any of Embodiments I-1 to
I-38, or a pharmaceutically acceptable salt thereof, for use in
promoting oxidative metabolism.
[0529] Embodiment I-52. Use of a compound of any of Embodiments I-1
to I-38, or a pharmaceutically acceptable salt thereof, for
treating, preventing, or reducing the risk of a disease or disorder
associated with
.alpha.-amino-.beta.-carboxymuconate-.epsilon.-semialdehyde
decarboxylase (ACMSD) dysfunction.
[0530] Embodiment I-53. Use of a compound of any of Embodiments I-1
to I-38, or a pharmaceutically acceptable salt thereof, for
treating, preventing, or reducing the risk of a disease or disorder
associated with reduced nicotinamide adenine dinucleotide
(NAD.sup.+) levels.
[0531] Embodiment I-54. Use of a compound of any of Embodiments I-1
to I-38, or a pharmaceutically acceptable salt thereof, for
treating, preventing, or reducing the risk of a disorder associated
with mitochondrial dysfunction.
[0532] Embodiment I-55. Use of a compound of any of Embodiments I-1
to I-38, or a pharmaceutically acceptable salt thereof, for
promoting oxidative metabolism.
[0533] Embodiment I-56. Use of a compound of any of Embodiments I-1
to I-38, or a pharmaceutically acceptable salt thereof, in the
manufacture of a medicament for treating, preventing, or reducing
the risk of a disease or disorder associated with
.alpha.-amino-.beta.-carboxymuconate-.epsilon.-semialdehyde
decarboxylase (ACMSD) dysfunction.
[0534] Embodiment I-57. Use of a compound of any of Embodiments I-1
to I-38, or a pharmaceutically acceptable salt thereof, in the
manufacture of a medicament for treating, preventing, or reducing
the risk of a disease or disorder associated with reduced
nicotinamide adenine dinucleotide (NAD.sup.+) levels.
[0535] Embodiment I-58. Use of a compound of any of Embodiments I-1
to I-38, or a pharmaceutically acceptable salt thereof, in the
manufacture of a medicament for treating, preventing, or reducing
the risk of a disorder associated with mitochondrial
dysfunction.
[0536] Embodiment I-59. Use of a compound of any of Embodiments I-1
to I-38, or a pharmaceutically acceptable salt thereof, in the
manufacture of a medicament for promoting oxidative metabolism.
[0537] Embodiment I-60. A method of treating, preventing, or
reducing the risk of a disease or disorder inhibited by
.alpha.-amino-.beta.-carboxymuconate-.epsilon.-semialdehyde
decarboxylase (ACMSD) comprising administering to the subject
suffering from or susceptible to developing the disease or disorder
a therapeutically effective amount of a pharmaceutical composition
of Embodiment I-39.
[0538] Embodiment I-61. A method of treating, preventing, or
reducing the risk of a disease or disorder associated with reduced
nicotinamide adenine dinucleotide (NAD.sup.+) levels comprising
administering to the subject suffering from or susceptible to
developing a disease or disorder associated with reduced NAD.sup.+
levels a therapeutically effective amount of a pharmaceutical
composition of Embodiment I-39.
[0539] Embodiment I-62. The method of any one of Embodiments I-60
to I-61, wherein the disease is chronic liver disease selected from
primary biliary cirrhosis (PBC), cerebrotendinous xanthomatosis
(CTX), primary sclerosing cholangitis (PSC), drug induced
cholestasis, intrahepatic cholestasis of pregnancy, parenteral
nutrition associated cholestasis (PNAC), bacterial overgrowth or
sepsis associated cholestasis, autoimmune hepatitis, chronic viral
hepatitis, alcoholic liver disease, nonalcoholic fatty liver
disease (NAFLD), nonalcoholic steatohepatitis (NASH), liver
transplant associated graft versus host disease, living donor
transplant liver regeneration, congenital hepatic fibrosis,
choledocholithiasis, granulomatous liver disease, intra- or
extrahepatic malignancy, Sjogren's syndrome, Sarcoidosis, Wilson's
disease, Gaucher's disease, hemochromatosis, and alpha
1-antitrypsin deficiency.
[0540] Embodiment I-63. A method of treating a disorder associated
with mitochondrial dysfunction comprising administering to the
subject suffering from or susceptible to developing a metabolic
disorder a therapeutically effective amount of a pharmaceutical
composition of Embodiment I-39.
[0541] Embodiment I-64. The method of Embodiment I-63, wherein said
disorder associated with mitochondrial dysfunction is an inherited
mitochondrial disease, a common metabolic disorder, a
neurodegenerative disease, an aging related disorder, a kidney
disorder, or a chronic inflammatory disease.
[0542] Embodiment I-65. The method of claim 64, wherein the common
metabolic disorder is obesity or type II diabetes.
[0543] Embodiment I-66. A method of promoting oxidative metabolism
comprising administering to the subject suffering from or
susceptible to developing a metabolic disorder a therapeutically
effective amount of a pharmaceutical composition of Embodiment
I-39.
[0544] Embodiment I-67. A pharmaceutical composition of Embodiment
I-39 for use as a medicament.
[0545] Embodiment I-68. A pharmaceutical composition of Embodiment
I-39 for use in treating, preventing, or reducing the risk of a
disease or disorder associated with reduced nicotinamide adenine
dinucleotide (NAD.sup.+) levels.
[0546] Embodiment I-69. A pharmaceutical composition of Embodiment
I-39 for use in for treating, preventing, or reducing the risk of a
disorder associated with mitochondrial dysfunction.
[0547] Embodiment I-70. A pharmaceutical composition of Embodiment
I-39 for use in promoting oxidative metabolism.
[0548] Embodiment I-71. Use of pharmaceutical composition of
Embodiment I-39 for treating, preventing, or reducing the risk of a
disease or disorder associated with
.alpha.-amino-.beta.-carboxymuconate-.epsilon.-semialdehyde
decarboxylase (ACMSD) dysfunction.
[0549] Embodiment I-72. Use of a pharmaceutical composition of
Embodiment I-39 for treating, preventing, or reducing the risk of a
disease or disorder associated with reduced nicotinamide adenine
dinucleotide (NAD.sup.+) levels.
[0550] Embodiment I-73. Use of pharmaceutical composition of
Embodiment I-39 for treating, preventing, or reducing the risk of a
disorder associated with mitochondrial dysfunction.
[0551] Embodiment I-74. Use pharmaceutical composition of
Embodiment I-39 for promoting oxidative metabolism.
[0552] Embodiment I-75. Use of pharmaceutical composition of
Embodiment I-39 in the manufacture of a medicament for treating,
preventing, or reducing the risk of a disease or disorder
associated with
.alpha.-amino-.beta.-carboxymuconate-.epsilon.-semialdehyde
decarboxylase (ACMSD) dysfunction.
[0553] Embodiment I-76. Use of pharmaceutical composition of
Embodiment I-39 in the manufacture of a medicament for treating,
preventing, or reducing the risk of a disease or disorder
associated with reduced nicotinamide adenine dinucleotide
(NAD.sup.+) levels.
[0554] Embodiment I-77. Use of pharmaceutical composition of
Embodiment I-39 in the manufacture of a medicament for treating,
preventing, or reducing the risk of a disorder associated with
mitochondrial dysfunction.
[0555] Embodiment I-78. Use of pharmaceutical composition of
Embodiment I-39 in the manufacture of a medicament for promoting
oxidative metabolism.
[0556] Embodiment II-1. A compound represented by Formula (II):
##STR00214##
[0557] or a pharmaceutically acceptable salt or tautomer thereof,
wherein:
[0558] X is H, S, SR.sup.2, NR.sup.2, NR.sup.2R.sup.2', O, OH,
OR.sup.h, F, Br, or Cl;
[0559] W is N or C; [0560] (i) when W is N, then: L is
--(C(R.sup.5).sub.2).sub.mCH.dbd.CH(C(R.sup.5).sub.2).sub.p,
##STR00215##
[0560] --(C(R.sup.5).sub.2).sub.mY.sup.1(C(R.sup.5).sub.2).sub.p--,
--(C(R.sup.5).sub.2).sub.mY.sup.1(C(R.sup.5).sub.2).sub.pcyclopropyl-,
--(C(R.sup.5).sub.2).sub.mY.sup.1CH.dbd.CH--,
--(C(R.sup.5).sub.2).sub.mNR.sup.3C.dbd.(O)(C(R.sup.5).sub.2).sub.p--,
--(C(R.sup.5).sub.2).sub.mphenyl(C(R.sup.5).sub.2).sub.p--,
--(C(R.sup.5).sub.2).sub.mpyridinyl(C(R.sup.5).sub.2).sub.p--, or
--(C(R.sup.5).sub.2).sub.mthiophenyl(C(R.sup.5).sub.2).sub.p--;
[0561] (ii) when W is C, then: L is
--(C(R.sup.5).sub.2).sub.mCH.dbd.CH(C(R.sup.5).sub.2).sub.p--,
--(C(R.sup.5).sub.2).sub.o--,
--(C(R.sup.5).sub.2).sub.mY.sup.1(C(R.sup.5).sub.2).sub.p,
##STR00216##
--(C(R.sup.5).sub.2).sub.m Y.sup.1CH.dbd.CH--,
--(C(R.sup.5).sub.2).sub.mC.dbd.(O)(CH.sub.2).sub.p,
--(C(R.sup.5).sub.2).sub.mC.dbd.(O)O(C(R.sup.5).sub.2).sub.p,
--(C(R.sup.5).sub.2).sub.mC.dbd.(O)NR.sup.3(C(R.sup.5).sub.2).sub.p,
--(C(R.sup.5).sub.2).sub.mNR.sup.3C.dbd.(O)(C(R.sup.5).sub.2).sub.p--,
--(C(R.sup.5).sub.2).sub.mphenyl(C(R.sup.5).sub.2).sub.p--,
--(C(R.sup.5).sub.2).sub.mpyridinyl(C(R.sup.5).sub.2).sub.p--, or
--(C(R.sup.5).sub.2).sub.mthiophenyl(C(R.sup.5).sub.2).sub.p--;
[0562] Y.sup.1 is O, NR.sup.4, or S(O).sub.q;
[0563] each Y.sup.2 is independently O, NH or S;
[0564] R.sup.1 is absent, C.sub.6-C.sub.10 arylene, heteroarylene,
or C.sub.3-C.sub.8cycloalkylene, wherein the heteroarylene
comprises one or two 5- to 7-membered rings and 1-4 heteroatoms
selected from N, O and S, and wherein the C.sub.6-C.sub.10 arylene,
heteroarylene, and C.sub.3-C.sub.8cycloalkylene are optionally
substituted with one to two R.sup.e;
[0565] R.sup.2 is H or C.sub.1-C.sub.4 alkyl;
[0566] R.sup.2' is H, C.sub.1-C.sub.4 alkyl, or C.sub.3-C.sub.7
cycloalkyl; or
[0567] R.sup.2 and R.sup.2' together with the nitrogen atom to
which they are attached form a 3- to 7-membered heterocycloalkyl
ring comprising 1-3 additional heteroatoms selected from N, O and
S;
[0568] R.sup.3 is H or C.sub.1-C.sub.4 alkyl;
[0569] R.sup.4 is H or C.sub.1-C.sub.4 alkyl;
[0570] each R.sup.5 is independently at each occurrence H or
C.sub.1-C.sub.4 alkyl;
[0571] each R.sup.6 is independently at each occurrence H or
C.sub.1-C.sub.4 alkyl;
[0572] R.sup.7 is H, A, B, or C;
[0573] A is --(C(R.sup.6).sub.2).sub.rCO.sub.2R.sup.x,
--Y.sup.2(C(R.sup.6).sub.2).sub.rCO.sub.2R.sup.x,
--(C(R.sup.6).sub.2).sub.rtetrazole,
--(C(R.sup.6).sub.2).sub.roxadiazolone,
--(C(R.sup.6).sub.2).sub.rtetrazolone,
--(C(R.sup.6).sub.2).sub.rthiadiazolol, --(C(R.sup.6).sub.2).sub.r
isoxazol-3-ol, --(C(R.sup.6).sub.2).sub.rP(O)(OH)OR.sup.x,
--(C(R.sup.6).sub.2).sub.rS(O).sub.2OH,
--(C(R.sup.6).sub.2).sub.rC(O)NHCN, or
--(C(R.sup.6).sub.2).sub.rC(O)NHS(O).sub.2alkyl, wherein
--(C(R.sup.6).sub.2).sub.rtetrazole,
--(C(R.sup.6).sub.2).sub.roxadiazolone,
--(C(R.sup.6).sub.2).sub.rtetrazolone,
--(C(R.sup.6).sub.2).sub.rthiadiazolol, --(C(R.sup.6).sub.2).sub.r
isoxazol-3-ol are optionally substituted with C.sub.1-C.sub.6
alkyl,
[0574] B is --(C(R.sup.6).sub.2).sub.rS(O).sub.2OC.sub.1-C.sub.4
alkyl, --O(C(R.sup.6).sub.2).sub.rS(O).sub.2OC.sub.1-C.sub.4 alkyl,
--Y.sup.2(C(R.sup.6).sub.2).sub.rC(O)NR.sup.gR.sup.g',
--Y.sup.2(C(R.sup.6).sub.2).sub.rS(O).sub.2NR.sup.gR.sup.g',
--(C(R.sup.6).sub.2).sub.rC(O)NR.sup.gR.sup.g',
--(C(R.sup.6).sub.2).sub.rS(O).sub.2NR.sup.gR.sup.g',
--(C(R.sup.6).sub.2).sub.rC(O)NHS(O).sub.2NR.sup.gR.sup.g',
--(C(R.sup.6).sub.2).sub.rCO.sub.2R.sup.w,
--(C(R.sup.6).sub.2).sub.rNH.sub.2CO.sub.2R.sup.x,
--(C(R.sup.6).sub.2).sub.rP(O)(OR.sup.x).sub.2,
--O(C(R.sup.6).sub.2).sub.rP(O)(OR.sup.x).sub.2,
--(C(R.sup.6).sub.2).sub.rS(O).sub.2OH,
--O(C(R.sup.6).sub.2).sub.rS(O).sub.2OH,
--(C(R.sup.6).sub.2).sub.rP(O).sub.2OR.sup.x, or
--O(C(R.sup.6).sub.2).sub.rP(O).sub.2OR.sup.x,
[0575] C is --(CH.sub.2).sub.rCN, --(CH.sub.2).sub.sOH, halogen,
--(C(R.sup.6).sub.2).sub.rC.sub.6-C.sub.10 aryl,
--(C(R.sup.6).sub.2).sub.rS--C.sub.6-C.sub.10 aryl,
--(C(R.sup.6).sub.2).sub.rheteroaryl,
--O(C(R.sup.6).sub.2).sub.rheteroaryl,
--O(C(R.sup.6).sub.2).sub.rheterocycloalkyl,
--O(C(R.sup.6).sub.2).sub.rOH, --OR.sup.y,
--(C(R.sup.6).sub.2).sub.rC(O)NHCN, --CH.dbd.CHCO.sub.2R.sup.x, or
--(C(R.sup.6).sub.2).sub.rC(O)NHS(O).sub.2C.sub.1-C.sub.4 alkyl,
wherein the aryl and heteroaryl are substituted with one to three
substituents each independently selected from C.sub.1-C.sub.6
alkyl, C.sub.1-C.sub.6 haloalkyl, halogen, and OH, and wherein the
heterocycloalkyl is substituted with one to two .dbd.O or
.dbd.S;
[0576] R.sup.c is H, C.sub.1-C.sub.6 alkyl, C.sub.1-C.sub.6
haloalkyl, halogen, --CN, --OR.sup.x, or --CO.sub.2R.sup.x;
[0577] R.sup.d is methyl, CF.sub.3, CR.sup.fF.sub.2,
--(C(R.sup.6).sub.2).sub.tC.sub.6-C.sub.10 aryl,
--(C(R.sup.6).sub.2).sub.t-5- or 6-membered heteroaryl,
--(C(R.sup.6).sub.2).sub.t-5- or 6-membered cycloalkyl, optionally
substituted C.sub.6-C.sub.10 aryl, optionally substituted 5- or
6-membered heteroaryl, or optionally substituted 5- or 6-membered
cycloalkyl;
[0578] each R.sup.e is independently at each occurrence
C.sub.1-C.sub.6 alkyl, C.sub.2-C.sub.6 alkenyl, C.sub.2-C.sub.6
alkynyl, halogen, C.sub.1-C.sub.6 haloalkyl, --NHR.sup.z, --OH, or
--CN;
[0579] R.sup.f is absent, H, or methyl;
[0580] R.sup.g is H, C.sub.1-C.sub.6 alkyl, OH,
--S(O).sub.2(C.sub.1-C.sub.6 alkyl), or S(O).sub.2N(C.sub.1-C.sub.6
alkyl).sub.2;
[0581] R.sup.g' is H, C.sub.1-C.sub.6 alkyl, C.sub.3-C.sub.7
cycloalkyl, 4- to 7-membered heterocycloalkyl ring comprising 1-3
heteroatoms selected from N, O and S, C.sub.6-C.sub.10 aryl, or 5-
to 7-membered heteroaryl comprising 1-3 heteroatoms selected from
N, O and S, wherein the alkyl is optionally substituted with one or
more substituents independently selected from halogen and --OH, and
wherein the cycloalkyl, heterocycloalkyl, aryl and heteroaryl are
optionally substituted with one or more substituents independently
selected from C.sub.1-C.sub.6 alkyl, halogen, and --OH;
[0582] R.sup.h is H, C.sub.1-C.sub.4 alkyl, or 3- to 7-membered
heterocycloalkyl ring comprising 1-3 heteroatoms selected from N, O
and S, wherein the alkyl is optionally substituted with one or more
substituents each independently selected from NH.sub.2,
C.sub.1-C.sub.4 alkylamino, C.sub.1-C.sub.4 dialkylamino, and
C(O)NH.sub.2; and wherein the heterocycloalkyl is optionally
substituted with one or more substituents each independently
selected from C.sub.1-C.sub.6 alkyl and C.sub.1-C.sub.6
haloalkyl;
[0583] R.sup.i is (i) --(CH.sub.2).sub.sOC(O)C.sub.1-C.sub.6 alkyl,
wherein the alkyl is substituted with one or more NH.sub.2; (ii)
(CH.sub.2CH.sub.2O).sub.nCH.sub.2CH.sub.2OH; or (iii)
C.sub.1-C.sub.6 alkyl substituted with one or more substituents
each independently selected from OH and 4- to 7-membered
heterocycloalkyl comprising 1 to 3 heteroatoms selected from O, N,
or S;
[0584] R.sup.i is absent, H, C.sub.1-C.sub.6 alkyl, or --CN;
[0585] each R.sup.x is independently at each occurrence H,
C.sub.1-C.sub.6 alkyl, or C.sub.6-C.sub.10 aryl;
[0586] each R.sup.y and R.sup.z is independently H, C.sub.1-C.sub.6
alkyl, or C.sub.1-C.sub.6 haloalkyl;
[0587] each m, p, q, r, and t is independently 0, 1 or 2;
[0588] n is 0, 1, 2, or 3;
[0589] s is 1 or 2;
[0590] o is 0, 1, 2, 3, or 4; and
[0591] represents a single bond or a double bond; and
[0592] provided that
[0593] when X is O; R.sup.f is H; W is C; R.sup.i is --CN; L is
--SCH.sub.2--; R.sup.1 is phenylene or pyridine; then R.sup.7 is
not --COOH;
[0594] when X is O; R.sup.f is H; W is C; R.sup.i is --CN; L is
--SCH.sub.2--; R.sup.1 is phenylene or pyridine; and R.sup.7 is
tetrazole; then R.sup.c is not H;
[0595] when X is O; R.sup.f is H; W is C; R.sup.i is --CN; L is
--S--C(R.sup.5).sub.2 or --SCH.sub.2CH.sub.2--; R.sup.1 is absent;
then R.sup.7 is not COOH or tetrazole;
[0596] when X is O, R.sup.f is H; W is N; R.sup.i is absent;
R.sup.d is methyl, optionally substituted 5- to 10-membered aryl,
optionally substituted 5- or 6-membered heteroaryl, or optionally
substituted 5- or 6-membered cycloalkyl; L is --SCH.sub.2-- or
--OCH.sub.2--; and R.sup.1 is phenylene; then R.sup.7 is not
--COOH, --CH.sub.2COOH,
##STR00217##
and
[0597] when X is O, R.sup.f is H, W is N, R.sup.i is absent, L is
--NHCH.sub.2--, --CH.sub.2NH--, or --NH--C(O)--, and R.sup.1 is
phenylene, then R.sup.d is not phenyl.
[0598] Embodiment II-2. A compound represented by Formula (I):
##STR00218##
[0599] or a pharmaceutically acceptable salt or tautomer
thereof,
[0600] wherein:
[0601] X is H, S, SR.sup.2, NR.sup.2, NR.sup.2R.sup.2', O, OH,
OR.sup.h, F, Br, or Cl;
[0602] W is N or C; [0603] (i) when W is N, then: [0604] L is
--(C(R.sup.5).sub.2).sub.mCH.dbd.CH(C(R.sup.5).sub.2).sub.p--,
[0604] ##STR00219##
--(C(R.sup.5).sub.2).sub.mY.sup.1(C(R.sup.5).sub.2).sub.p--,
--(C(R.sup.5).sub.2).sub.mY.sup.1(C(R.sup.5).sub.2).sub.p-cyclopropyl-,
--(C(R.sup.5).sub.2).sub.mY.sup.ICH.dbd.CH--,
--(C(R.sup.5).sub.2).sub.mNR.sup.3C.dbd.(O)(C(R.sup.5).sub.2).sub.p--,
--(C(R.sup.5).sub.2).sub.mphenyl(C(R.sup.5).sub.2).sub.p--,
--(C(R.sup.5).sub.2).sub.mpyridinyl(C(R.sup.5).sub.2).sub.p--, or
--(C(R.sup.5).sub.2).sub.mthiophenyl(C(R.sup.5).sub.2).sub.p--;
[0605] (ii) when W is C, then: [0606] L is
--(C(R.sup.5).sub.2).sub.mCH.dbd.CH(C(R.sup.5).sub.2).sub.p--,
--(C(R.sup.5).sub.2).sub.o--,
--(C(R.sup.5).sub.2).sub.mY.sup.1(C(R.sup.5).sub.2).sub.p--,
[0606] ##STR00220## --(C(R.sup.5).sub.2).sub.m Y.sup.ICH.dbd.CH--,
--(C(R.sup.5).sub.2).sub.mC.dbd.(O)(CH.sub.2).sub.p,
--(C(R.sup.5).sub.2).sub.mC.dbd.(O)O(C(R.sup.5).sub.2).sub.p--,
--(C(R.sup.5).sub.2).sub.mC.dbd.(O)NR.sup.3(C(R.sup.5).sub.2).sub.p--,
--(C(R.sup.5).sub.2).sub.mNR.sup.3C.dbd.(O)(C(R.sup.5).sub.2).sub.p--,
--(C(R.sup.5).sub.2).sub.mphenyl(C(R.sup.5).sub.2).sub.p--,
--(C(R.sup.5).sub.2).sub.mpyridinyl(C(R.sup.5).sub.2).sub.p--, or
--(C(R.sup.5).sub.2).sub.mthiophenyl(C(R.sup.5).sub.2).sub.p--;
[0607] Y.sup.1 is O, NR.sup.4, or S(O).sub.q;
[0608] each Y.sup.2 is independently O, NH or S;
[0609] R.sup.1 is absent or C.sub.6-C.sub.10 arylene or
heteroarylene, wherein the heteroarylene comprises one or two 5- to
7-membered rings and 1-4 heteroatoms selected from N, O and S, and
wherein the C.sub.6-C.sub.10 arylene or heteroarylene are
optionally substituted with one to two R.sup.e;
[0610] R.sup.2 is H or C.sub.1-C.sub.4 alkyl;
[0611] R.sup.2' is H, C.sub.1-C.sub.4 alkyl, or C.sub.3-C.sub.7
cycloalkyl; or
[0612] R.sup.2 and R.sup.2' together with the nitrogen atom to
which they are attached form a 3- to 7-membered heterocycloalkyl
ring comprising 1-3 additional heteroatoms selected from N, O and
S;
[0613] R.sup.3 is H or C.sub.1-C.sub.4 alkyl;
[0614] R.sup.4 is H or C.sub.1-C.sub.4 alkyl;
[0615] each R.sup.5 is independently at each occurrence H or
C.sub.1-C.sub.4 alkyl;
[0616] each R.sup.6 is independently at each occurrence H or
C.sub.1-C.sub.4 alkyl;
[0617] R.sup.7 is H, A, B, or C;
[0618] A is --(C(R.sup.6).sub.2).sub.rCO.sub.2R.sup.x,
--Y.sup.2(C(R.sup.6).sub.2).sub.rCO.sub.2R.sup.x,
--(CH.sub.2).sub.rtetrazole, --(CH.sub.2).sub.roxadiazolone,
--(CH.sub.2).sub.rtetrazolone, --(CH.sub.2).sub.rthiadiazolol,
--(CH.sub.2).sub.r isoxazol-3-ol,
--(CH.sub.2).sub.rP(O)(OH)OR.sup.x, --(CH.sub.2).sub.rS(O).sub.2OH,
--(CH.sub.2).sub.rC(O)NHCN, or
--(CH.sub.2).sub.rC(O)NHS(O).sub.2alkyl, wherein
--(CH.sub.2).sub.rtetrazole, --(CH.sub.2).sub.roxadiazolone,
--(CH.sub.2).sub.rtetrazolone, --(CH.sub.2).sub.rthiadiazolol,
--(CH.sub.2).sub.r isoxazol-3-ol are optionally substituted with
C.sub.1-C.sub.6 alkyl,
[0619] B is --(C(R.sup.6).sub.2).sub.rS(O).sub.2OC.sub.1-C.sub.4
alkyl, --O(C(R.sup.6).sub.2).sub.rS(O).sub.2OC.sub.1-C.sub.4 alkyl,
--Y.sup.2(C(R.sup.6).sub.2).sub.rC(O)NR.sup.gR.sup.g',
--Y.sup.2(C(R.sup.6).sub.2).sub.rS(O).sub.2NR.sup.gR.sup.g',
--(CH.sub.2).sub.rC(O)NR.sup.gR.sup.g',
--(CH.sub.2).sub.rS(O).sub.2NR.sup.gR.sup.g',
--(CH.sub.2).sub.rC(O)NHS(O).sub.2NR.sup.gR.sup.g',
--(C(R.sup.6).sub.2).sub.rCO.sub.2R.sup.i,
--(C(R.sup.6).sub.2).sub.rNH.sub.2CO.sub.2R.sup.x,
--(C(R.sup.6).sub.2).sub.rP(O)(OR.sup.x).sub.2,
--O(C(R.sup.6).sub.2).sub.rP(O)(OR.sup.x).sub.2,
--(C(R.sup.6).sub.2).sub.rS(O).sub.2OH,
--O(C(R.sup.6).sub.2).sub.rS(O).sub.2OH,
--(C(R.sup.6).sub.2).sub.rP(O).sub.2OR.sup.x, or
--O(C(R.sup.6).sub.2).sub.rP(O).sub.2OR.sup.x,
[0620] C is --(CH.sub.2).sub.rCN, --(CH.sub.2).sub.sOH, halogen,
--(C(R.sup.6).sub.2).sub.rC.sub.6-C.sub.10 aryl,
--(C(R.sup.6).sub.2).sub.rS--C.sub.6-C.sub.10 aryl,
--(C(R.sup.6).sub.2).sub.rheteroaryl,
--O(C(R.sup.6).sub.2).sub.rheteroaryl,
--O(C(R.sup.6).sub.2).sub.rheterocycloalkyl,
--O(C(R.sup.6).sub.2).sub.rOH, --OR.sup.y,
--(C(R.sup.6).sub.2).sub.rC(O)NHCN, --CH.dbd.CHCO.sub.2R.sup.x, or
--(C(R.sup.6).sub.2).sub.rC(O)NHS(O).sub.2C.sub.1-C.sub.4 alkyl,
wherein the aryl and heteroaryl are substituted with one to three
substituents each independently selected from C.sub.1-C.sub.6
alkyl, C.sub.1-C.sub.6 haloalkyl, halogen, and OH, and wherein the
heterocycloalkyl is substituted with one to two .dbd.O or
.dbd.S;
[0621] R.sup.c is H, C.sub.1-C.sub.6 alkyl, C.sub.1-C.sub.6
haloalkyl, halogen, --CN, --OR.sup.x, or --CO.sub.2R.sup.x;
[0622] R.sup.d is methyl, CF.sub.3, CR.sup.fF.sub.2,
--(C(R.sup.6).sub.2).sub.tC.sub.6-C.sub.10 aryl,
--(C(R.sup.6).sub.2).sub.t-5- or 6-membered heteroaryl,
--(C(R.sup.6).sub.2).sub.t-5- or 6-membered cycloalkyl, optionally
substituted C.sub.6-C.sub.10 aryl, optionally substituted 5- or
6-membered heteroaryl, or optionally substituted 5- or 6-membered
cycloalkyl;
[0623] each R.sup.e is independently at each occurrence
C.sub.1-C.sub.6 alkyl, C.sub.2-C.sub.6 alkenyl, C.sub.2-C.sub.6
alkynyl, halogen, C.sub.1-C.sub.6 haloalkyl, --NHR.sup.z, --OH, or
--CN;
[0624] R.sup.f is absent, H, or methyl;
[0625] R.sup.g is H, C.sub.1-C.sub.6 alkyl, OH,
--S(O).sub.2(C.sub.1-C.sub.6 alkyl), or S(O).sub.2N(C.sub.1-C.sub.6
alkyl).sub.2;
[0626] R.sup.g' is H, C.sub.1-C.sub.6 alkyl, C.sub.3-C.sub.7
cycloalkyl, 4- to 7-membered heterocycloalkyl ring comprising 1-3
heteroatoms selected from N, O and S, C.sub.6-C.sub.10 aryl, or 5-
to 7-membered heteroaryl comprising 1-3 heteroatoms selected from
N, O and S, wherein the alkyl is optionally substituted with one or
more substituents independently selected from halogen and --OH, and
wherein the cycloalkyl, heterocycloalkyl, aryl and heteroaryl are
optionally substituted with one or more substituents independently
selected from C.sub.1-C.sub.6 alkyl, halogen, and --OH;
[0627] R.sup.h is H, C.sub.1-C.sub.4 alkyl, or 3- to 7-membered
heterocycloalkyl ring comprising 1-3 heteroatoms selected from N, O
and S, wherein the alkyl is optionally substituted with one or more
substituents each independently selected from NH.sub.2,
C.sub.1-C.sub.4 alkylamino, C.sub.1-C.sub.4 dialkylamino, and
C(O)NH.sub.2; and wherein the heterocycloalkyl is optionally
substituted with one or more substituents each independently
selected from C.sub.1-C.sub.6 alkyl and C.sub.1-C.sub.6
haloalkyl;
[0628] R.sup.i is (i) --(CH.sub.2).sub.sOC(O)C.sub.1-C.sub.6 alkyl,
wherein the alkyl is substituted with one or more NH.sub.2; (ii)
(CH.sub.2CH.sub.2O).sub.nCH.sub.2CH.sub.2OH; or (iii)
C.sub.1-C.sub.6 alkyl substituted with one or more substituents
each independently selected from OH and 4- to 7-membered
heterocycloalkyl comprising 1 to 3 heteroatoms selected from O, N,
or S;
[0629] R.sup.j is absent, H, C.sub.1-C.sub.6 alkyl, or --CN;
[0630] each R.sup.x is independently at each occurrence H,
C.sub.1-C.sub.6 alkyl, or C.sub.6-C.sub.10 aryl;
[0631] each R.sup.y and R.sup.z is independently H, C.sub.1-C.sub.6
alkyl, or C.sub.1-C.sub.6 haloalkyl;
[0632] each m, p, q, r, and t is independently 0, 1 or 2;
[0633] n is 0, 1, 2, or 3;
[0634] s is 1 or 2;
[0635] o is 0, 1, 2, 3, or 4; and
[0636] represents a single bond or a double bond; and
[0637] provided that
[0638] when X is O; R.sup.f is H; W is C; R.sup.j is --CN; L is
--SCH.sub.2--; R.sup.1 is phenylene or pyridine; then R.sup.7 is
not --COOH;
[0639] when X is O; R.sup.f is H; W is C; R.sup.j is --CN; L is
--SCH.sub.2--; R.sup.1 is phenylene or pyridine; and R.sup.7 is
tetrazole; then R.sup.c is not H;
[0640] when X is O; R.sup.f is H; W is C; R.sup.j is --CN; L is
--S--C(R.sup.5).sub.2 or --SCH.sub.2CH.sub.2--; R.sup.1 is absent;
then R.sup.7 is not COOH or tetrazole;
[0641] when X is O, R.sup.f is H; W is N; R.sup.i is absent;
R.sup.d is methyl, optionally substituted 5- to 10-membered aryl,
optionally substituted 5- or 6-membered heteroaryl, or optionally
substituted 5- or 6-membered cycloalkyl; L is --SCH.sub.2-- or
--OCH.sub.2--; and R.sup.1 is phenylene; then R.sup.7 is not
--COOH, --CH.sub.2COOH,
##STR00221##
and
[0642] when X is O, R.sup.f is H, W is N, R.sup.j is absent, L is
--NHCH.sub.2--, --CH.sub.2NH--, or --NH--C(O)--, and R.sup.1 is
phenylene, then R.sup.d is not phenyl.
[0643] Embodiment II-3. The compound of Embodiment II-1 or II-2,
wherein X is O, OH, OR.sup.h, F, Br, or Cl.
[0644] Embodiment II-4. The compound of Embodiment II-1 or II-2,
wherein X is H, S, SR.sup.2, NR.sup.2, or NR.sup.2R.sup.2'.
[0645] Embodiment II-5. The compound of any one of Embodiments II-1
to II-4, wherein R.sup.f is absent.
[0646] Embodiment II-6. The compound of any one of Embodiments II-1
to II-4, wherein R.sup.f is H or methyl.
[0647] Embodiment II-7. The compound of any one of Embodiments II-1
to II-6, wherein W is N.
[0648] Embodiment II-8. The compound of Embodiment II-7, wherein
R.sup.i is absent.
[0649] Embodiment II-9. The compound of any one Embodiments II-1 to
II-6, wherein W is C.
[0650] Embodiment II-10. The compound of Embodiment II-9, wherein
R.sup.i is H, C.sub.1-C.sub.6 alkyl, or --CN.
[0651] Embodiment II-11. The compound of Embodiments II-9 or II-10,
wherein R.sup.i is --CN.
[0652] Embodiment II-12. The compound of any one of Embodiments
II-1 to II-11, wherein R.sup.c is C.sub.1-C.sub.6 alkyl, --CN, or
halogen.
[0653] Embodiment II-13. The compound of any one of Embodiments
II-1 to II-12, wherein R.sup.c is --CN or halogen.
[0654] Embodiment II-14. The compound of any one of Embodiments
II-1 to II-12, wherein R.sup.c is --CN.
[0655] Embodiment II-15. The compound of any one of Embodiments
II-1 to II-14, wherein R.sup.d is methyl.
[0656] Embodiment II-16. The compound of any one of Embodiments
II-1 to II-14, wherein R.sup.d is optionally substituted 5- to
10-membered aryl.
[0657] Embodiment II-17. The compound of any one of Embodiments
II-1 to II-14, wherein R.sup.d is optionally substituted 5- or
6-membered heteroaryl.
[0658] Embodiment II-18. The compound of any one of Embodiments
II-1 to II-14, wherein R.sup.d is optionally substituted 5- or
6-membered cycloalkyl.
[0659] Embodiment II-19. The compound of any one of Embodiments
II-1 to II-14, wherein R.sup.d is methyl, cyclohexyl, pyridinyl,
thiazolyl, phenyl, or thienyl.
[0660] Embodiment II-20. The compound of any one of Embodiments
II-1 to II-14, wherein R.sup.d is methyl, cyclohexyl, pyridinyl,
thiazolyl, thienyl, or optionally substituted phenyl.
[0661] Embodiment II-21. The compound of any one of Embodiments
II-1 to II-14, wherein R.sup.d is methyl.
[0662] Embodiment II-22. The compound of any one of Embodiments
II-1 to II-14, wherein R.sup.d is --CF.sub.3.
[0663] Embodiment II-23. The compound of any one of Embodiments
II-1 to II-14, wherein R.sup.d is CR.sup.fF.sub.2.
[0664] Embodiment II-24. The compound of any one of Embodiments
II-1 to II-14, wherein R.sup.d is
--(C(R.sup.6).sub.2).sub.rC.sub.6-C.sub.10 aryl,
--(C(R.sup.6).sub.2).sub.r-5- or 6-membered heteroaryl,
--(C(R.sup.6).sub.2).sub.r-5- or 6-membered cycloalkyl.
[0665] Embodiment II-25. The compound of any one of Embodiments
II-1 to II-14, wherein R.sup.d is
--(C(R.sup.6).sub.2).sub.rC.sub.6-C.sub.10 aryl.
[0666] Embodiment II-26. The compound of any one of Embodiments
II-1 to II-25, wherein L is
--(C(R.sup.5).sub.2).sub.mY.sup.1(C(R.sup.5).sub.2).sub.p--.
[0667] Embodiment II-27. The compound of Embodiment II-26, wherein
Y.sup.1 is S.
[0668] Embodiment II-28. The compound of any one of Embodiments
II-1 to II-25, wherein L is
--(C(R.sup.5).sub.2).sub.mNR.sup.3C.dbd.(O)(C(R.sup.5).sub.2).sub.p--
or
--(C(R.sup.5).sub.2).sub.mY.sup.1(C(R.sup.5).sub.2).sub.pcyclopropyl-.
[0669] Embodiment II-29. The compound of any one of Embodiments
II-1 to II-28, wherein R.sup.1 is C.sub.6-C.sub.10 arylene.
[0670] Embodiment II-30. The compound of any one of Embodiments
II-1 to II-28, wherein R.sup.1 is heteroarylene.
[0671] Embodiment II-31. The compound of any one of Embodiments
II-1 to II-28, wherein R.sup.1 is absent.
[0672] Embodiment II-32. The compound of any one of Embodiments
II-1 to II-31, wherein R.sup.7 is A.
[0673] Embodiment II-33. The compound of Embodiment II-32, wherein
A is --(C(R.sup.6).sub.2).sub.rCO.sub.2R.sup.x or
--(CH.sub.2).sub.rtetrazole, wherein the
--(CH.sub.2).sub.rtetrazole is optionally substituted with
C.sub.1-C.sub.6 alkyl.
[0674] Embodiment II-34. The compound of any one of Embodiments
II-1 to II-31, wherein R.sup.7 is B.
[0675] Embodiment II-35. The compound of Embodiment II-32, wherein
B is --(CH.sub.2).sub.rC(O)NR.sup.gR.sup.g', or
--(CH.sub.2).sub.rS(O).sub.2NR.sup.gR.sup.g',
[0676] Embodiment II-36. The compound of any one of Embodiments
II-1 to II-31, wherein R.sup.7 is C.
[0677] Embodiment II-37. The compound of Embodiment II-32, wherein
C is --(CH.sub.2).sub.rCN, --(CH.sub.2).sub.sOH, or
--(C(R.sup.6).sub.2).sub.rC.sub.6-C.sub.10 aryl, wherein the aryl
is substituted with one to three substituents each independently
selected from C.sub.1-C.sub.6 alkyl, C.sub.1-C.sub.6 haloalkyl,
halogen, and OH.
[0678] Embodiment II-38. A compound, or a pharmaceutically
acceptable salt or tautomer thereof, selected from the group
consisting of
TABLE-US-00005 Cpd No. Structure I-1 ##STR00222## I-2 ##STR00223##
I-3 ##STR00224## I-4 ##STR00225## I-5 ##STR00226## I-6 ##STR00227##
I-7 ##STR00228## I-8 ##STR00229## I-9 ##STR00230## I-10
##STR00231## I-11 ##STR00232## I-12 ##STR00233## I-13 ##STR00234##
I-14 ##STR00235## I-15 ##STR00236## I-16 ##STR00237## I-17
##STR00238## I-18 ##STR00239## I-19 ##STR00240## I-20 ##STR00241##
I-21 ##STR00242## I-22 ##STR00243## I-23 ##STR00244## I-24
##STR00245## I-25 ##STR00246## I-26 ##STR00247## I-27 ##STR00248##
I-28 ##STR00249## I-29 ##STR00250## I-30 ##STR00251## I-31
##STR00252## I-32 ##STR00253## I-33 ##STR00254## I-34 ##STR00255##
I-35 ##STR00256## I-36 ##STR00257##
[0679] Embodiment II-39. A compound, or a pharmaceutically
acceptable salt or tautomer thereof, selected from the group
consisting of
##STR00258## ##STR00259## ##STR00260##
[0680] Embodiment II-40. A compound, or a pharmaceutically
acceptable salt or tautomer thereof, selected from the group
consisting of
##STR00261## ##STR00262## ##STR00263## ##STR00264## ##STR00265##
##STR00266## ##STR00267## ##STR00268## ##STR00269## ##STR00270##
##STR00271##
[0681] Embodiment II-41. A pharmaceutical composition comprising a
compound of any one of Embodiments II-1 to II-40, or a
pharmaceutically acceptable salt thereof, and at least one of a
pharmaceutically acceptable carrier, diluent, or excipient.
[0682] Embodiment II-42. The pharmaceutical composition according
to Embodiment II-41, which comprises one or more further
therapeutic agents.
[0683] Embodiment II-43. A method of treating, preventing, or
reducing the risk of a disease or disorder inhibited by
.alpha.-amino-.beta.-carboxymuconate-.epsilon.-semialdehyde
decarboxylase (ACMSD) comprising administering to the subject
suffering from or susceptible to developing the disease or disorder
a therapeutically effective amount of one or more compounds of any
one of Embodiments II-1 to II-40, or a pharmaceutically acceptable
salt thereof.
[0684] Embodiment II-44. A method of treating, preventing, or
reducing the risk of a disease or disorder associated with reduced
nicotinamide adenine dinucleotide (NAD.sup.+) levels comprising
administering to the subject suffering from or susceptible to
developing a disease or disorder associated with reduced NAD.sup.+
levels a therapeutically effective amount of one or more compounds
of any one of Embodiments II-1 to II-40, or a pharmaceutically
acceptable salt thereof.
[0685] Embodiment II-45. The method of any one of Embodiments II-43
to II-44, wherein the disease is chronic liver disease selected
from primary biliary cirrhosis (PBC), cerebrotendinous
xanthomatosis (CTX), primary sclerosing cholangitis (PSC), drug
induced cholestasis, intrahepatic cholestasis of pregnancy,
parenteral nutrition associated cholestasis (PNAC), bacterial
overgrowth or sepsis associated cholestasis, autoimmune hepatitis,
chronic viral hepatitis, alcoholic liver disease, nonalcoholic
fatty liver disease (NAFLD), nonalcoholic steatohepatitis (NASH),
liver transplant associated graft versus host disease, living donor
transplant liver regeneration, congenital hepatic fibrosis,
choledocholithiasis, granulomatous liver disease, intra- or
extrahepatic malignancy, Sjogren's syndrome, Sarcoidosis, Wilson's
disease, Gaucher's disease, hemochromatosis, and alpha
1-antitrypsin deficiency.
[0686] Embodiment II-46. A method of treating a disorder associated
with mitochondrial dysfunction comprising administering to the
subject suffering from or susceptible to developing a metabolic
disorder a therapeutically effective amount of one or more
compounds of any one of Embodiments II-1 to II-40, or a
pharmaceutically acceptable salt thereof, that increases
intracellular nicotinamide adenine dinucleotide (NAD.sup.+).
[0687] Embodiment II-47. The method of Embodiment II-46, wherein
said disorder associated with mitochondrial dysfunction is an
inherited mitochondrial disease, a common metabolic disorder, a
neurodegenerative disease, an aging related disorder, a kidney
disorder, or a chronic inflammatory disease.
[0688] Embodiment II-48. The method of Embodiment II-47, wherein
the common metabolic disorder is obesity or type II diabetes.
[0689] Embodiment II-49. A method of promoting oxidative metabolism
comprising administering to the subject suffering from or
susceptible to developing a metabolic disorder a therapeutically
effective amount of one or more compounds of any one of Embodiments
II-1 to II-40, or a pharmaceutically acceptable salt thereof, that
increases intracellular nicotinamide adenine dinucleotide
(NAD.sup.+).
[0690] Embodiment II-50. A compound of any of Embodiments II-1 to
II-40, or a pharmaceutically acceptable salt thereof, for use as a
medicament.
[0691] Embodiment II-51. A compound of any of Embodiments II-1 to
II-40, or a pharmaceutically acceptable salt thereof, for use in
treating, preventing, or reducing the risk of a disease or disorder
associated with reduced nicotinamide adenine dinucleotide
(NAD.sup.+) levels.
[0692] Embodiment II-52. A compound of any of Embodiments II-1 to
II-40, or a pharmaceutically acceptable salt thereof, for use in
for treating, preventing, or reducing the risk of a disorder
associated with mitochondrial dysfunction.
[0693] Embodiment II-53. A compound of any of Embodiments II-1 to
II-40, or a pharmaceutically acceptable salt thereof, for use in
promoting oxidative metabolism.
[0694] Embodiment II-54. Use of a compound of any of Embodiments
II-1 to II-40, or a pharmaceutically acceptable salt thereof, for
treating, preventing, or reducing the risk of a disease or disorder
associated with
.alpha.-amino-.beta.-carboxymuconate-.epsilon.-semialdehyde
decarboxylase (ACMSD) dysfunction.
[0695] Embodiment II-55. Use of a compound of any of Embodiments
II-1 to II-40, or a pharmaceutically acceptable salt thereof, for
treating, preventing, or reducing the risk of a disease or disorder
associated with reduced nicotinamide adenine dinucleotide
(NAD.sup.+) levels.
[0696] Embodiment II-56. Use of a compound of any of Embodiments
II-1 to II-40, or a pharmaceutically acceptable salt thereof, for
treating, preventing, or reducing the risk of a disorder associated
with mitochondrial dysfunction.
[0697] Embodiment II-57. Use of a compound of any of Embodiments
II-1 to II-40, or a pharmaceutically acceptable salt thereof, for
promoting oxidative metabolism.
[0698] Embodiment II-58. Use of a compound of any of Embodiments
II-1 to II-40, or a pharmaceutically acceptable salt thereof, in
the manufacture of a medicament for treating, preventing, or
reducing the risk of a disease or disorder associated with
.alpha.-amino-.beta.-carboxymuconate-.epsilon.-semialdehyde
decarboxylase (ACMSD) dysfunction.
[0699] Embodiment II-59. Use of a compound of any of Embodiments
II-1 to II-40, or a pharmaceutically acceptable salt thereof, in
the manufacture of a medicament for treating, preventing, or
reducing the risk of a disease or disorder associated with reduced
nicotinamide adenine dinucleotide (NAD.sup.+) levels.
[0700] Embodiment II-60. Use of a compound of any of Embodiments
II-1 to II-40, or a pharmaceutically acceptable salt thereof, in
the manufacture of a medicament for treating, preventing, or
reducing the risk of a disorder associated with mitochondrial
dysfunction.
[0701] Embodiment II-61. Use of a compound of any of Embodiments
II-1 to II-40, or a pharmaceutically acceptable salt thereof, in
the manufacture of a medicament for promoting oxidative
metabolism.
[0702] Embodiment II-62. A method of treating, preventing, or
reducing the risk of a disease or disorder inhibited by
.alpha.-amino-.beta.-carboxymuconate-.epsilon.-semialdehyde
decarboxylase (ACMSD) comprising administering to the subject
suffering from or susceptible to developing the disease or disorder
a therapeutically effective amount of a pharmaceutical composition
of Embodiment II-41.
[0703] Embodiment II-63. A method of treating, preventing, or
reducing the risk of a disease or disorder associated with reduced
nicotinamide adenine dinucleotide (NAD.sup.+) levels comprising
administering to the subject suffering from or susceptible to
developing a disease or disorder associated with reduced NAD.sup.+
levels a therapeutically effective amount of a pharmaceutical
composition of Embodiment II-41.
[0704] Embodiment II-64. The method of any one of Embodiments II-62
to II-63, wherein the disease is chronic liver disease selected
from primary biliary cirrhosis (PBC), cerebrotendinous
xanthomatosis (CTX), primary sclerosing cholangitis (PSC), drug
induced cholestasis, intrahepatic cholestasis of pregnancy,
parenteral nutrition associated cholestasis (PNAC), bacterial
overgrowth or sepsis associated cholestasis, autoimmune hepatitis,
chronic viral hepatitis, alcoholic liver disease, nonalcoholic
fatty liver disease (NAFLD), nonalcoholic steatohepatitis (NASH),
liver transplant associated graft versus host disease, living donor
transplant liver regeneration, congenital hepatic fibrosis,
choledocholithiasis, granulomatous liver disease, intra- or
extrahepatic malignancy, Sjogren's syndrome, Sarcoidosis, Wilson's
disease, Gaucher's disease, hemochromatosis, and alpha
1-antitrypsin deficiency.
[0705] Embodiment II-65. A method of treating a disorder associated
with mitochondrial dysfunction comprising administering to the
subject suffering from or susceptible to developing a metabolic
disorder a therapeutically effective amount of a pharmaceutical
composition of Embodiment II-41.
[0706] Embodiment II-66. The method of Embodiment II-65, wherein
said disorder associated with mitochondrial dysfunction is an
inherited mitochondrial disease, a common metabolic disorder, a
neurodegenerative disease, an aging related disorder, a kidney
disorder, or a chronic inflammatory disease.
[0707] Embodiment II-67. The method of Embodiment II-66, wherein
the common metabolic disorder is obesity or type II diabetes.
[0708] Embodiment II-68. A method of promoting oxidative metabolism
comprising administering to the subject suffering from or
susceptible to developing a metabolic disorder a therapeutically
effective amount of a pharmaceutical composition of Embodiment
II-41.
[0709] Embodiment II-69. A pharmaceutical composition of Embodiment
II-41 for use as a medicament.
[0710] Embodiment II-70. A pharmaceutical composition of Embodiment
II-41 for use in treating, preventing, or reducing the risk of a
disease or disorder associated with reduced nicotinamide adenine
dinucleotide (NAD.sup.+) levels.
[0711] Embodiment II-71. A pharmaceutical composition of Embodiment
II-41 for use in for treating, preventing, or reducing the risk of
a disorder associated with mitochondrial dysfunction.
[0712] Embodiment II-72. A pharmaceutical composition of Embodiment
II-41 for use in promoting oxidative metabolism.
[0713] Embodiment II-73. Use of pharmaceutical composition of
Embodiment II-41 for treating, preventing, or reducing the risk of
a disease or disorder associated with
.alpha.-amino-.beta.-carboxymuconate-s-semialdehyde decarboxylase
(ACMSD) dysfunction.
[0714] Embodiment II-74. Use of a pharmaceutical composition of
Embodiment II-41 for treating, preventing, or reducing the risk of
a disease or disorder associated with reduced nicotinamide adenine
dinucleotide (NAD.sup.+) levels.
[0715] Embodiment II-75. Use of pharmaceutical composition of
Embodiment II-41 for treating, preventing, or reducing the risk of
a disorder associated with mitochondrial dysfunction.
[0716] Embodiment II-76. Use pharmaceutical composition of
Embodiment II-41 for promoting oxidative metabolism.
[0717] Embodiment II-77. Use of pharmaceutical composition of
Embodiment II-41 in the manufacture of a medicament for treating,
preventing, or reducing the risk of a disease or disorder
associated with
.alpha.-amino-.beta.-carboxymuconate-.epsilon.-semialdehyde
decarboxylase (ACMSD) dysfunction.
[0718] Embodiment II-78. Use of pharmaceutical composition of
Embodiment II-41 in the manufacture of a medicament for treating,
preventing, or reducing the risk of a disease or disorder
associated with reduced nicotinamide adenine dinucleotide
(NAD.sup.+) levels.
[0719] Embodiment II-79. Use of pharmaceutical composition of
Embodiment II-41 in the manufacture of a medicament for treating,
preventing, or reducing the risk of a disorder associated with
mitochondrial dysfunction.
[0720] Embodiment II-80. Use of pharmaceutical composition of
Embodiment II-41 in the manufacture of a medicament for promoting
oxidative metabolism.
EXAMPLES
[0721] All percentages and ratios used herein, unless otherwise
indicated, are by weight. Other features and advantages of the
present disclosure will become apparent from the different
examples. The provided examples illustrate different components and
methodology useful in practicing the present disclosure. Generally
speaking, the disclosure extends to any novel one, or any novel
combination, of the features disclosed in this specification
(including the accompanying claims and drawings). The examples do
not limit the claimed disclosure. Thus, features, integers,
characteristics, compounds or chemical moieties described in
conjunction with a particular aspect, embodiment or example of the
disclosure are to be understood to be applicable to any other
aspect, embodiment or example described herein, unless incompatible
therewith. Based on the present disclosure the skilled artisan can
identify and employ other components and methodology useful for
practicing the present disclosure. Moreover, unless stated
otherwise, any feature disclosed herein may be replaced by an
alternative feature serving the same or a similar purpose.
[0722] The Disclosure will now be described by way of example only
with reference to the Examples below:
EXEMPLIFICATION
Compound Preparation
General Methods and Materials
[0723] All chemicals were purchased from Sigma-Aldrich, Alfa Aesar.
.sup.1H NMR spectra were recorded at 200 and 400 MHz and .sup.13C
NMR spectra were recorded at 100.6 and 50.3 MHz by using deuterated
solvents indicated below. TLC were performed on aluminium backed
silica plates (silica gel 60 F254). All the reactions were
performed under nitrogen atmosphere using distilled solvents. All
tested compounds were found to have >95% purity determined by
HPLC analysis. HPLC-grade water was obtained from a tandem
Milli-Ro/Milli-Q apparatus. The analytical HPLC measurements were
made on a Shimadzu LC-20AProminence equipped with a CBM-20A
communication bus module, two LC-20AD dual piston pumps, a SPD-M20A
photodiode array detector and a Rheodyne 7725i injector with a 20
.mu.L stainless steel loop.
[0724] Abbreviations used in the following examples and elsewhere
herein are: [0725] Ac.sub.2O acetic anhydride [0726] AcOH acetic
acid [0727] AIBN Azobisisobutyronitrile [0728] atm atmosphere
[0729] br broad [0730] DIPEA N,N-diisopropylethylamine [0731] DCM
dichloromethane [0732] DME dimethoxyethane [0733] DMF
N,N-dimethylformamide [0734] DMSO dimethyl sulfoxide [0735] BPO
Dibenzoylperoxide [0736] EDC
N-(3-Dimethylaminopropyl)-N'-ethylcarbodiimide hydrochloride [0737]
ESI electrospray ionization [0738] EtOAc ethyl acetate [0739]
EtO.sub.2 diethyl ether [0740] EtOH ethanol [0741]
EtO.sup.-Na.sup.+ sodium ethoxide [0742] Et.sub.3NH.sup.+Cl.sup.-
triethylamine hydrochloride [0743] h hour(s) [0744] HPLC
high-performance liquid chromatography [0745] LCMS liquid
chromatography-mass spectrometry [0746] m multiplet [0747] Mel
methyl iodide [0748] MeOH methanol [0749] MHz megahertz [0750] min
minutes [0751] MS molecular sieves [0752] MTBE
2-methoxy-2-methylpropane [0753] MW microwave [0754] NBS
N-bromosuccinamide [0755] NMR nuclear magnetic resonance [0756] PET
petroleum ether [0757] ppm parts per million [0758] p-TSA
para-toluenesulfonic acid [0759] r.t. room temperature [0760] TLC
thin layer chromatography
Example 1: Intermediate 1.4.
4-Oxo-6-thiophen-2-yl-2-thioxo-1,2,3,4-tetrahydro-pyrimidine-5-carbonitri-
le
##STR00272##
[0762] To a stirred solution of compound 1.1 (0.96 g, 8.8 mmol),
1.2 (672 mg, 8.8 mmol) and 1.3 (1 g, 0.83 mL) in ethanol (55 mL)
was added K.sub.2CO.sub.3 (1.57 g, 11.44 mmol). Stirring was
continued at reflux overnight. The yellowish solid formed was
collected after cooling, taken up with hot water and filtered
again. The aqueous phase was acidified to pH1, the precipitate was
filtered and dried under reduced pressure. The title compound 1.4
was obtained as a yellowish solid (1 g, 4.25 mmol). Yield 49%.
.sup.1H NMR (200 MHz, DMSO-d.sub.6) .delta. 7.22 (m, 1H), 7.68 (m,
1H), 7.85 (d, J=4.8 Hz, 1H), 8.05 (s, 1H).
Example 2: Intermediate 2.2. Sodium;
6-oxo-4-trifluoromethyl-1,6-dihydro-pyrimidine-2-thiolate
##STR00273##
[0764] Sodium (0.35 g, 16.29 mmol) was dissolved in abs. EtOH (25
mL) under N.sub.2 atmosphere. To the resulting solution ethyl
trifluoroacetoacetate 2.1 (1.59 mL, 10.86 mmol) and thiourea 1.2
(0.91 g, 11.94 g) were added. The mixture was stirred and refluxed
for 4 h. Once cooled at room temperature the obtained precipitate
was collected by filtration under vacuum and washed with cold EtOH
(2.times.5 mL), to afford (1.34 g, 6.14 mmol) of intermediate 2.2.
Yield 38%. MS-ESI(-) m/z: 194.8 [M-H].
Example 3: Intermediate 3.3.
2-Mercapto-6-oxo-4-phenyl-1,6-dihydro-pyridine-3-carbonitrile
##STR00274##
[0766] To a stirred solution of KOH (0.58 g, 10.41 mmol) in abs.
EtOH (20 mL), ethyl 3-oxo-3-phenyl-propionate 3.1 (1.80 mL, 10.41
mmol) and 2-cyanothioacetamide 3.2 (1.04 g, 10.41 mmol) were added,
and the resulting mixture was stirred and refluxed for 3 hours.
Then it was cooled at room temperature and concentrated under
reduced pressure. The crude was poured in H.sub.2O (20 mL) and
washed with AcOEt (2.times.15 mL). The organic phase was acidified
un to pH=2 by adding aq. HCl 37%, and the resulting precipitate was
collected by filtration under vacuum and washed with H.sub.2O
(2.times.5 mL). The solid was then tritured with AcMe, to give
intermediate 3.3 (0.49 g, 2.14 mmol) as a yellowish solid. Yield
21%. MS-ESI(-) m/z: 227.3 [M-H].sup.-
Example 4: Intermediate 4.2.
4-Benzyl-2-mercapto-6-oxo-1,6-dihydro-pyrimidine-5-carbonitrile
##STR00275##
[0768] To a solution of phenyl acetaldehyde 4.1 (1.5 g, 16.65
mmol), ethylcyanoacetate 1.1 (1.41 g, 16.65 mmol) and thiourea 1.2
(950 mg, 16.65 mmol) in EtOH (35 mL) was added K.sub.2CO.sub.3 (2.2
g, 21.6 mmol). Stirring was continued at reflux 16 h. The mixture
was cooled to r.t. The white solid was collected, dissolved in
water. The pH was adjusted to 3 by the addition of 3N HCl. The
aqueous phase was extracted with EtOAc (3.times.50 mL). The
combined organic phase was washed with brine and dried over
anhydrous Na.sub.2SO.sub.4. The title intermediate 4.2 (800 mg,
3.28 mmol) was obtained as a light yellow solid. Yield: 20%.
.sup.1H NMR (200 MHz, DMSO-d.sub.6) .delta. 3.93 (s, 2H), 7.26-7.41
(m, 5H), 13.15 (brs, 1H).
Example 5: Intermediate 5.2.
2-Mercapto-6-oxo-4-thiophen-2-yl-1,6-dihydro-pyridine-3-carbonitrile
##STR00276##
[0770] To a stirred solution of KOH (0.28 g, 5.04 mmol) in abs.
EtOH (10 mL), ethyl 3-oxo-3-thiophen-2-yl-propionate 5.1 (0.77 mL,
5.04 mmol) and 2-cyanothioacetamide 3.2 (0.50 g, 5.04 mmol) were
added, and the resulting mixture was stirred at reflux for 8 hours.
Then it was cooled at room temperature and the precipitate formed
was collected by filtration under vacuum and washed with EtOH
(2.times.5 mL), to give intermediate 5.2 (0.17 g, 0.72) as a
yellowish solid. Yield 12%. MS-ESI(-) m/z: 233.3 [M-H].sup.-.
Example 6: Intermediate 6.4.
6-Mercapto-2-oxo-4-thiophen-2-yl-1,2-dihydro-pyridine-3,5-dicarbonitrile
##STR00277##
[0771] Step 1: 2-Cyano-3-thiophen-2-yl-acrylic acid ethyl ester
(6.1)
[0772] To a solution of thiophene-2-carboxaldehyde 1.3 (1 g, 8.9
mmol), ethylcyanoacetate 1.1 (0.94 mL, 8.9 mmol) in EtOH (20 mL)
was added piperidine (3 drops). Stirring was continued ar r.t. 16
h. The solvent was removed under vacuo. The crude was taken up with
water, extracted with EtOAc (3.times.50 mL). The organic phase was
collected, washed with brine and dried over anhydrous
Na.sub.2SO.sub.4. The title intermediate 6.1 (1.3 g, 6.27 mmol) was
obtained as a white solid. Yield 70%.
Step 2:
6-Mercapto-2-oxo-4-thiophen-2-yl-1,2-dihydro-pyridine-3,5-dicarbon-
itrile (6.2)
[0773] To a solution of intermediate 6.1 (1.2 g, 5.79 mmol) in EtOH
(15 mL) was added piperidine (4 drops). Stirring was continued at
reflux 16 h. Upon cooling a red precipitate was formed. The
precipitate was collected, washed with cold EtOH, and dried under
vacuo. The title intermediate 6.2 (640 mg, 2.46 mmol) as a red
powder. Yield 42%. .sup.1H NMR (200 MHz, DMSO-d.sub.6) .delta.
7.24-7.27 (m, 1H), 7.53-7.55 (m, 1H), 7.94-7.95 (m, 1H), 13.0 (brs,
1H).
Example 7: Intermediate 7.1. Potassium;
3-cyano-6-oxo-4-trifluoromethyl-1,6-dihydro-pyridine-2-thiolate
##STR00278##
[0775] To a stirred solution of KOH (0.91 g, 16.29 mmol) in abs.
EtOH (32 mL), ethyl trifluoroacetoacetate 2.1 (2.38 mL, 16.29 mmol)
and 2-cyanothioacetamide 3.2 (1.63 g, 16.29 mmol) were added, and
the resulting mixture was stirred and refluxed for 7 hours. Then it
was cooled at room temperature and left to stand overnight. The
copious precipitate thus formed was collected by filtration under
vacuum and washed with EtOH (2.times.5 mL), to give intermediate
7.1 (2.01 g, 7.78 mmol) as a white solid. Yield 48%. MS-ESI(-) m/z:
218.9 [M-H].sup.-
Example 8: Intermediate 8.3. (3-Bromomethyl-phenyl)-acetic acid
ethyl ester
##STR00279##
[0776] Step 1: m-Tolyl-acetic acid ethyl ester (8.2)
[0777] To a solution of 8.1 (15 g, 99.88 mmol) in EtOH (absolute)
(400 mL) was added HCl (conc.) (0.3 mL, 9.9 mmol) and stirring was
continued at reflux for 4 h. The volatiles were removed under
reduced pressure. The crude was taken up with DCM (200 mL) dried
over Na.sub.2SO.sub.4 and evaporated under reduced pressure. The
title compound 8.2 was obtained as a colorless oil (17 g, 95.39
mmol). Yield 96%. .sup.1H NMR (200 MHz, CDCl.sub.3) .delta. 1.28
(t, J=7.1 Hz, 3H), 2.37 (s, 2H), 3.6 (s, 2H), 4.18 (q, J=7.11 Hz,
2H), 7.20-7.35 (m, 4H). GC/MS m/z 178.1 (M+).
Step 2: (3-Bromomethyl-phenyl)-acetic acid ethyl ester (8.3)
[0778] NBS (10.1 g, 58.9 mmol) and BPO (70%) (68 mg, 0.28 mmol)
were added to a solution of intermediate 8.2 (10 g, 56.11 mmol) in
CH.sub.3CN (300 mL). Stirring was continued at reflux for 4h. The
volatile were removed under reduced pressure. The crude residue was
partitioned between EtOAc (300 mL) and a saturated NaHCO.sub.3
aqueous solution (300 mL). The organic phase was collected and
dried over Na.sub.2SO.sub.4. The crude product was purified by
flash chromatography (dry load) eluting with PET/Et.sub.2O from 2%
to 4% for product. The title compound 8.3 (10 g, 38.89 mmol) was
obtained as a yellowish oil. Yield 66%. .sup.1H NMR (200 MHz,
CDCl.sub.3) .delta. 1.27 (t, J=7.1 Hz, 3H), 3.62 (s, 2H), 4.17 (q,
J=7.13 Hz, 2H), 4.50 (s, 2H), 7.09-7.13 (m, 3H), 7.21-7.28 (m,
1H).
Example 9: Intermediate 9.3. 3-Bromomethyl-benzamide
##STR00280##
[0779] Step 1: 3-Methyl-benzamide (9.2)
[0780] A solution of compound 9.1 (1.54 mL, 12.8 mmol) and
K.sub.2CO.sub.3 (707 mg, 5.12 mmol) in H.sub.2O (5 mL) was heated
under microwave irradiation at 130.degree. C., 200 psi, 200 W for
20 minutes. Upon cooling, the resulting white precipitate was
collected and dried under reduced pressure to afford the title
compound 9.2 as white crystals (870 mg, 6.4 mmol). Yield 50%. GC-MS
(m/z) 135.1 (M+).
Step 2: 3-Bromomethyl-benzamide (9.3)
[0781] NBS (434.6 mg, 2.4 mmol) and BPO (70%) (8 mg, 0.022 mmol)
were added to a solution of intermediate 9.2 (300 mg, 2.22 mmol) in
CH.sub.3CN (20 mL). Stirring was continued at reflux for 4h. The
volatile were removed under reduced pressure. The crude product was
partitioned between EtOAc (300 mL) and a saturated NaHCO.sub.3
aqueous solution (300 mL). The organic phase was collected and
dried over Na.sub.2SO.sub.4. The title compound 9.3 (250 mg, 1.16
mmol) was obtained as a yellowish solid. Yield 53%.
Example 10: Intermediate 10.4.
3'-Bromomethyl-3,5-difluoro-4-methoxy-biphenyl
##STR00281##
[0782] Step 1: 3,5-Difluoro-4-methoxy-3'-methyl-biphenyl (10.3)
[0783] To a solution of compound 10.1 (0.18 mL, 1.33 mmol) in DME
(15 mL) was added palladium tetrakis (50 mg, 0.039 mmol). Stirring
was continued at r.t. for 5 min. m-Tolyl boronic acid 10.2 (202 mg,
1.35 mmol) and K.sub.2CO.sub.3 (745 mg, 3.56 mmol) were added in
turn. Stirring was continued at reflux for 4h. The solvent was
removed under reduced pressure. The crude residue was taken up in
water and extracted with DCM (3.times.20 ml). The organic phase was
washed with brine, dried over Na.sub.2SO.sub.4, filtered and
concentrated under reduced pressure. Pure title compound 10.3 (282
mg, 1.22 mmol) was obtained as a colorless oil and it was used for
the next step without further purification. Yield 91%. .sup.1H NMR
(400 MHz, CDCl.sub.3) .delta. 2.43 (s, 3H), 4.04 (s, 3H), 7.14 (d,
J=9.3, 2H), 7.32-7.33 (m, 4H).
Step 2: 3'-Bromomethyl-3,5-difluoro-4-methoxy-biphenyl (10.4)
[0784] To a solution of the intermediate 10.3 (260 mg, 1.1 mmol) in
CH.sub.3CN (15 mL) was added BPO (4 mg, 0.0055 mmol) and NBS (210
mg, 1.22 mmol). Stirring was continued at reflux overnight. The
solvent was removed under reduced pressure. The reaction
partitioned between NaHCO.sub.3(ss) and DCM. The organic phase was
washed with brine and dried over Na.sub.2SO.sub.4. The crude was
purified by flash chromatography, eluting with PET/Et.sub.2O
affording the title compound 10.4 (250 mg, 0.77 mmol) as a yellow
oil. Yield 72%. .sup.1H NMR (400 MHz, CDCl.sub.3) .delta. 4.06 (d,
J=3.7 Hz, 3H), 4.55 (s, 2H), 7.14 (d, J=6.2 Hz, 1H), 7.16 (d, J=6.1
Hz, 1H), 7.41-7.47 (m, 3H), 7.54 (s, 1H).
Example 11: Intermediate 11.2. (3-Bromomethyl-phenyl)-acetic
acid
##STR00282##
[0786] To a suspension of compound 11.1 (750 mg, 5 mmol) in
CCl.sub.4 (15 mL) was added AIBN (41 mg, 0.25 mmol) and NBS (933.7
mg, 5.24 mmol). Stirring was continued at reflux overnight. The
solvent was removed under reduced pressure. The reaction was taken
up with water, extracted with EtOAc (3.times.20 mL) washed with
brine, and dried over Na.sub.2SO.sub.4. The crude was purified by
flash chromatography, eluting with CH.sub.2Cl.sub.2/MeOH (3% for
product) affording the title intermediate 11.2 (800 mg, 3.49 mmol)
as a white solid. Yield 70%.
[0787] GC/MS (m/z) 227.9 (M+).
Example 12: Intermediate 12.2.
[3-(4-Chloro-5-cyano-6-thiophen-2-yl-pyrimidin-2-ylsulfanylmethyl)-phenyl-
]-acetic acid
##STR00283##
[0788] Step 1:
[3-(5-Cyano-6-oxo-4-thiophen-2-yl-1,6-dihydro-pyrimidin-2-ylsulfanylmethy-
l)-phenyl]-acetic acid (12.1)
[0789] To a stirred suspension of intermediate 1.4 (500 mg, 2.12
mmol) and DIPEA (0.4 mL, 2.12 mmol) in DMSO (5 mL) was added
intermediate 11.2 (487 mg, 2.12 mmol). Stirring was continued
overnight at room temperature. The crude reaction mixture was
poured into water and the resulting aqueous mixture was washed with
EtOAc, acidified to pH 3, and extracted with EtOAc (3.times.50 mL).
The title intermediate 12.1 was obtained (200 mg, 0.52 mmol) as a
pure yellowish solid after flash chromatography purification
eluting with CH.sub.2Cl.sub.2/MeOH (10% for product) and shredding
with a mixture of EtO.sub.2/Acetone. Yield 25%. .sup.1H NMR (400
MHz, DMSO-d.sub.6) .delta. 3.49 (s, 2H), 4.53 (s, 2H), 7.16 (d,
J=6.8 Hz, 1H), 7.26 (t, J=7.2 Hz, 1H), 7.36 (m, 3H), 8.05 (d, J=4.4
Hz, 1H), 8.27 (s, 1H), 12.13 (s, 1H); .sup.13C NMR (100 MHz,
DMSO-d.sub.6) .delta. 34.3, 40.9, 88.5, 116.8, 127.6, 128.9, 129.1,
129.8, 130.4, 131.9, 135.2, 135.8, 137.0, 139.9, 159.1, 161.6,
165.7, 172.9. HPLC 95.8%.
Step 2:
[3-(4-Chloro-5-cyano-6-thiophen-2-yl-pyrimidin-2-ylsulfanylmethyl)-
-phenyl]-acetic acid (12.2)
[0790] A mixture of intermediate 12.1 (300 mg, 0.78 mmol) and
POCl.sub.3 (6 ml) were heated at 80.degree. C. 4 h. The crude
reaction mixture was then poured in ice. The resulting yellow
precipitate was collected and dried under reduced pressure
affording the title intermediate 12.2 (250 mg, 0.62 mmol) as a
yellowish solid. Yield 79%. .sup.1H NMR (400 MHz, DMSO-d.sub.6)
.delta. 3.53 (s, 2H), 4.50 (s, 2H), 7.16 (d, J=7.5 Hz, 1H), 7.27
(t, J=7.4 Hz, 1H), 7.36-7.39 (m, 3H), 8.13 (d, J=4.9 Hz, 1H), 8.3
(d, J=3.9 Hz, 1H), 12.25 (brs, 1H). .sup.13C NMR (100 MHz,
DMSO-d.sub.6) .delta. 35.1, 40.9, 97.7, 115.5, 127.6, 128.8, 129,
130.2, 130.4, 133.3, 135.7, 136, 137, 138.6, 160.3, 163.2, 172.9,
174.
Example 13: Intermediate 13.3.
[3-(4-Chloro-5-cyano-6-thiophen-2-yl-pyrimidin-2-ylsulfanylmethyl)-benzoi-
c acid
##STR00284##
[0791] Step 1:
3-(5-Cyano-6-oxo-4-thiophen-2-yl-1,6-dihydro-pyrimidin-2-ylsulfanylmethyl-
)-benzoic acid (13.2)
[0792] To a stirred suspension of intermediate 1.4 (250 mg, 1.06
mmol) and K.sub.2CO.sub.3 (440 mg, 3.18 mmol) in CH.sub.3CN (15 mL)
was added 3-(chloromethyl)benzoic acid 13.1 (180 mg, 1.06 mmol).
Stirring was continued overnight at reflux. The volatiles were then
removed under reduced pressure. The crude product was taken up in
water, washed with EtOAc, acidified to pH 1, and extracted with
EtOAc (3.times.50 mL). Shredding with hot acetone afforded the
title intermediate 13.2 (45 mg, 0.12 mmol) as a yellowish solid.
Yield 12%. .sup.1H NMR (400 MHz, DMSO-d.sub.6) .delta. 4.62 (s,
2H), 7.33 (t, J=4.3 Hz, 1H), 7.44 (t, J=7.6 Hz, 1H), 7.72 (d, J=7.5
Hz, 1H), 7.82 (d, J=7.5 Hz, 1H), 8.05 (m, 2H), 8.26 (d, J=3.8 Hz,
1H), 12.99 (s, 1H); .sup.13C NMR (100 MHz, DMSO) .delta. 33.9,
88.7, 116.5, 128.8, 129.3, 129.9, 130.2, 131.5, 132.1, 133.7,
135.4, 137.9, 139.7, 159.0, 161.2, 165.3, 167.4. HPLC: 97.2%
Step 2:
3-(4-Chloro-5-cyano-6-thiophen-2-yl-pyrimidin-2-ylsulfanylmethyl)--
benzoic acid (13.3)
[0793] A mixture of intermediate 13.2 (300 mg, 0.81 mmol) and
POCl.sub.3 (6 ml) were heated at 80.degree. C. for 4 h. The
reaction mixture was then poured into ice. The resulting yellow
precipitate was collected and purified by flash chromatography
eluting with DCM/MeOH (3% for product) to provide intermediate 13.3
(120 mg, 0.3 mmol) as a yellowish solid. Yield 79%. .sup.13C NMR
(100 MHz, DMSO-d.sub.6) .delta. 33.8, 88.7, 116.5, 128.8, 129.3,
129.9, 130.2, 131.4, 132.1, 133.7, 135.5, 137.9, 139.6, 159, 161.1,
165.2, 167.3;
Example 14: Intermediate 14.2. 3-Bromomethyl-benzonitrile
##STR00285##
[0795] To a solution of compound 14.1 (2 mL, 17.07 mmol) in
CCl.sub.4 was added a mixture of NBS (2.9 g, 17.1 mmol) and BPO (16
mg, 0.06 mmol). Stirring was continued at reflux for 16 h and the
reaction was then allowed to warm to rt. The resulting solid was
collected, washed with CCl.sub.4, and dried under reduced pressure.
The title compound 14.2 was obtained as a white solid (2.84 g, 14.5
mmol). Yield 85%. GC-) 196.9 (M+).
Example 15: Intermediate 15.1. 2-(3-Bromomethyl-phenyl)-ethanol
##STR00286##
[0797] To a solution of intermediate 11.2 (500 mg, 2.17 mmol) in
THF (10 mL) at 0.degree. C. was added BH.sub.3-THF (1M in THF, 2.8
mL) dropwise. The mixture was stirred at 0.degree. C. for 1 h and
then at r.t. for 12h. The mixture was diluted with THF/H.sub.2O
(1:1 v:v, 15 mL) and washed with saturated aq. K.sub.2CO.sub.3. The
phases were separated and the aqueous layer was extracted with
EtOAc (3.times.20 mL). The combined organic phase was washed with
brine and dried over Na.sub.2SO.sub.4. Flash chromatography
purification of the crude product (eluting with DCM/MeOH) afforded
the title intermediate 15.1 (400 mg, 1.85 mmol) as a white solid.
Yield 85%. GC/MS (m/z) 214 (M+).
Example 16: Intermediate 16.2. 2
(3-Bromomethyl-phenyl)-acetonitrile
##STR00287##
[0799] NBS (338 mg, 1.9 mmol) and BPO (70%) (28.7 mg, 0.11 mmol)
were added to a solution of intermediate 16.1 (0.5 mL, 2.37 mmol)
in CH.sub.3CN (15 mL). Stirring was continued at reflux for 4h. The
volatiles were removed under reduced pressure. The crude product
was partitioned between EtOAc (100 mL) and a saturated NaHCO.sub.3
aqueous solution (100 mL). The organic phase was collected and
dried over Na.sub.2SO.sub.4. The title compound 16.2 (250 mg, 1.18
mmol) was obtained as a yellowish solid after flash chromatography
purification (eluting with PET/EtOAc). Yield 50%.
Example 17: Intermediate 17.3.
5-(3-Bromomethyl-phenyl)-2-methyl-2H-tetrazole
##STR00288##
[0800] Step 1: 5-m-Tolyl-2H-tetrazole (17.1)
[0801] A mixture of compound 14.1 (1.02 mL, 8.54 mmol), NaN.sub.3
(832 mg, 12.8 mmol) and Et.sub.3N--HCl (1.76 g, 12.8 mmol) in
toluene (20 mL) was heated at reflux for 4 h. The solvent was then
removed under reduced pressure. The crude product was poured into
water and the resulting aqueous solution was acidified to pH 1 with
3N HCl and extracted with EtOAc (3.times.20 mL). The organic phase
was washed with brine, dried over Na.sub.2SO.sub.4, and
concentrated under reduced pressure. The title compound 17.1 (1.22
g, 7.6 mmol) was obtained as a white solid. Yield 89%. .sup.1H NMR
(200 MHz, DMSO-d.sub.6) .delta. 2.39 (s, 3H), 7.39 (m, 1H), 7.48
(t, J=7.58 Hz, 1H), 7.80 (s, 1H), 7.85 (m, 1H); GC/MS (m/z) 160.1
(M+).
Step 2: 2-Methyl-5-m-tolyl-2H-tetrazole (17.2)
[0802] To a solution of intermediate 17.1 (1 g, 6.2 mmol) in water
(5 mL) and NaOH (500 mg, 12.5 mmol) was added a solution of Mel
(0.38 mL, 6.1 mmol) in acetone (10 mL). Stirring was continued at
reflux for 6h. The solvent was then removed under reduced pressure
and the resulting residue was taken up in EtOAc and H.sub.2O. The
organic layer was separated, dried over Na.sub.2SO.sub.4 and
evaporated to dryness in vacuo. Purification of the crude product
afforded the title intermediate 17.2 (500 mg, 2.87 mmol) as a white
solid. Yield 46%. .sup.1H NMR (400 MHz, CDCl.sub.3) .delta. 4.37
(s, 3H), 7.26-7.39 (m, 1H), 7.35-7.39 (m, 1H), 7.91-7.96 (m,
2H).
Step 3: 5-(3-Bromomethyl-phenyl)-2-methyl-2H-tetrazole (17.3)
[0803] To a suspension of compound 17.2 (200 mg, 1.15 mmol) in
CH.sub.3CN (15 mL) was added BPO (21 mg, 0.057 mmol) and NBS (163.5
mg, 0.92 mmol). Stirring was continued at 92.degree. C. overnight.
The solvent was removed under reduced pressure. The reaction
mixture was taken up in water, extracted with EtOAc (3.times.20
mL), washed with brine, and dried over Na.sub.2SO.sub.4. The crude
product was purified by flash chromatography eluting with
CH.sub.2Cl.sub.2/MeOH (7% for product) to afford the title compound
17.3 (261 mg, 1.03 mmol) as a white solid. Yield 90%. .sup.1H NMR
(400 MHz, CDCl.sub.3) .delta. 4.41 (s, 3H), 4.56 (s, 2H), 7.46-7.52
(m, 1H), 8.07-8.09 (m, 1H), 8.19 (s, 1H).
Example 18: Intermediate 18.1.
5-(3-Bromomethyl-phenyl)-1H-tetrazole
##STR00289##
[0805] To a suspension of compound 17.1 (300 mg, 1.87 mmol) in
CH.sub.3CN (15 mL) was added AIBN (31 mg, 0.18 mmol) and NBS (333
mg, 1.87 mmol). Stirring was continued at reflux overnight. The
solvent was removed under reduced pressure. The reaction was taken
up with water, extracted with EtOAc (3.times.20 mL) washed with
brine, and dried over Na.sub.2SO.sub.4. The crude was purified by
flash chromatography, eluting with CH.sub.2Cl.sub.2/MeOH (7% for
product) affording the title compound 18.1 (150 mg, 0.62 mmol) as a
light yellow solid. Yield 34%.
Example 19: Intermediate 19.5. 3-Bromomethyl-benzenesulfonamide
##STR00290##
[0806] Step 1: 3-Chdorosulfonyl-bezoic acid (19.2)
[0807] A mixture of compound 19.1 (1 g, 8.13 mmol) and
chlorosulfonic acid (4 mL) was stirred at 125.degree. C. for 2 h.
The mixture was poured into ice water dropwise. The resulting solid
was collected, solubilized in EtOAc, and washed with water
(3.times.20 mL). The organic layer was dried over Na.sub.2SO4 and
evaporated under reduced pressure. The title intermediate 19.2
(1.19 g, 5.39 mmol) was obtained as a white solid. Yield 65%.
.sup.1H NMR (400 MHz, DMSO-d.sub.6) .delta. 7.45 (t, J=7.69 Hz,
1H), 7.65 (d, J=7.8 Hz, 1H), 7.86 (d, J=7.6 Hz, 1H), 8.1 (s, 1H),
13.9 (brs, 1H).
Step 2: 3-Sulfamoyl-benzoic acid (19.3)
[0808] To a cold solution of 25% NH40H (10 mL) was added
portionwise intermediate 19.2 (1.10 g, 5.39 mmol). Stirring was
continued at rt for 2 h and the resulting mixture was concentrated.
The crude product was suspended in water (4 mL) and 37% HCl
solution was then added dropwise to the mixture. The resulting
precipitate was collected and dried under reduced pressure to
afford the title intermediate 19.3 (943 mg, 4.6 mmol) as a white
solid. Yield 87%. .sup.1H NMR (400 MHz, DMSO-d.sub.6) .delta. 7.50
(brs, 2H), 7.71 (t, J=7.78 Hz, 1H), 8.04 (d, J=7.8 Hz, 1H), 8.13
(d, J=7.7 Hz, 1H), 8.38 (s, 1H), 13.4 (brs, 1H).
Step 3: 3-Hydroxymethyl-benzenesulfonamide (19.4)
[0809] To a stirred solution of intermediate 19.3 (940 mg, 4.67
mmol) was added dropwise at 0.degree. C. BH.sub.3-THF complex (14
mL, 14.01 mmol) and stirring was continued for 4 h at rt. The
reaction mixture was then cooled to 0.degree. C., and quenched by
the dropwise addition of MeOH. After 15 min, a 3N solution of HCl
(37 mL) was added to the mixture and the volatiles were removed
under reduced pressure. The aqueous phase was extracted with EtOAc
(3.times.20 mL). The combined organic phase was washed with brine
and dried over Na.sub.2SO.sub.4 to afford the title intermediate
19.4 (785 mg, 4.2 mmol) as a colorless oil. Yield 89%. .sup.1H NMR
(400 MHz, DMSO-d.sub.6) .delta. 4.51 (s, 2H), 7.34 (s, 2H), 7.5 (d,
J=5 Hz, 2H), 7.68 (t, J=5.2 Hz, 1H), 7.8 (s, 1H).
Step 4: 3-Bromomethyl-benzenesulfonamide (19.5)
[0810] To a stirred suspension of intermediate 19.4 (200 mg, 1.07
mmol) in DCM (3.5 mL) was added PBr.sub.3 and stirring was
continued at 20.degree. C. for 16h. Water was then added carefully
to the mixture and the phases were separated. The aqueous phase was
extracted with DCM (2.times.20 mL). The combined organic layers
were washed with brine and dried over Na.sub.2SO.sub.4 to afford
the title intermediate 19.5 (120 mg, 0.47 mmol) as a colorless oil.
Yield 45%. .sup.1H NMR (400 MHz, CDCl.sub.3) .delta. 4.53 (s, 2H),
7.54 (t, J=10.7 Hz, 1H), 7.69 (d, J=7.6 Hz, 1H), 7.88 (d, J=7.8 Hz,
1H), 7.97 (s, 1H).
Example 20: Intermediate 20.2.
4-Benzyl-2-mercapto-6-oxo-1,6-dihydro-pyridine-3-carbonitrile
##STR00291##
[0812] To a solution of intermediate 20.1 (1.2 g, 6.24 mmol) and
potassium tert butoxide (764 mg, 6.24 mmol) in DMF (15 mL) was
added compound 3.2 (31 mg, 0.18 mmol).
[0813] Stirring was continued at 85.degree. C. overnight. The
reaction was poured water, The pH was acidified to 5 by the
addition of AcOH followed by washing with EtOAc (3.times.20 mL).
Then, pH was brought to 3 by the addition of 3N HCl solution. The
aqueous phase was extracted with EtOAc (3.times.30 mL). The organic
phase was washed with brine and dried over anhydrous Na.sub.2SO4.
The title compound (600 mg, 2.47 mmol) was obtained as light yellow
solid. Yield 40%. .sup.1H NMR (400 MHz, DMSO-d.sub.6) .delta. 3.63
(s, 2H), 5.81 (s, 1H), 7.17-7.36 (m, 5H), 13.1 (brs, 1H).
Example 21:
[3-(5-Cyano-6-oxo-4-thiophen-2-yl-1,6-dihydro-pyrimidin-2-ylsulfanylmethy-
l)-phenyl]-acetic acid ethyl ester (Compound I-1)
##STR00292##
[0815] To a stirred suspension of intermediate 1.4 (2.13 g, 9.1
mmol) and K.sub.2CO.sub.3 (1.88 g, 13.6 mmol) in CH.sub.3CN (80 mL)
was added intermediate 8.3 (2.45 g, 9.52 mmol) and stirring was
continued at a gentle reflux for 16h. The solvent was then removed
under reduced pressure. The crude product was taken up in water and
the resulting aqueous solution was neutralized with 3N HCl
solution. The resulting pale yellow solid was collected, washed
with ice cold water, and dried under reduced pressure. The title
compound I-1 (3.1 g, 7.46 mmol) was obtained as a grey solid after
trituration with Et.sub.2O. Yield 82%. .sup.1H NMR (400 MHz,
DMSO-d.sub.6) .delta. 1.15 (d, J=7.03 Hz, 3H), 3.62 (s, 2H), 4.03
(q, J=7.16 Hz, 2H), 4.55 (s, 2H), 7.17 (d, J=7.1 Hz, 1H), 7.28 (t,
J=7.7 Hz, 1H), 7.38 (m, 3H), 8.1 (d, J=4.7 Hz, 1H), 8.29 (d, J=3.35
Hz, 1H). .sup.13C NMR (100 MHz, DMSO-d.sub.6) .delta. 14.4, 34.2,
60.7, 88.6, 116.6, 127.8, 129.1, 129.1, 129.9, 130.3, 132.1, 135.2,
135.5, 137.1, 139.7, 159.1, 161.1, 165.3, 171.4. HPLC>97.9%.
Example 22:
3-(5-Cyano-6-oxo-4-thiophen-2-yl-1,6-dihydro-pyrimidin-2-ylsulfanylmethyl-
)-benzamide (Compound I-2)
##STR00293##
[0817] To a stirred suspension of intermediate 1.4 (182 mg, 0.78
mmol) and intermediate 9.3 (200 mg, 0.65 mmol) in CH.sub.3CN (20
mL) was added K.sub.2CO.sub.3 (119 mg, 0.86 mmol) and stirring was
continued at a gentle reflux for 16h. The volatiles were removed
under reduced pressure. The crude product was taken up in water,
and the resulting aqueous mixture was acidified to pH 3 with a 3N
HCl solution and extracted with EtOAc (3.times.20 mL). The combined
organic phases were washed with brine and dried over
Na.sub.2SO.sub.4 to afford the title compound I-2 (120 mg, 0.24
mmol) as a yellowish solid after trituration with hot Et.sub.2O.
Yield 42%. .sup.1H NMR (400 MHz, DMSO-d.sub.6) .delta. 4.61 (s,
2H), 7.3 (m, 1H), 7.41 (m, 2H), 7.63 (d, J=7.12 Hz, 1H), 7.76 (d,
J=7.3 Hz, 1H), 7.97 (s, 2H), 8.01 (d, J=4.47 Hz, 1H), 8.3 (d, J=2.8
Hz, 1H). .sup.13C NMR (100 MHz, DMSO-d.sub.6) .delta. 34.5, 88.1,
117.1, 127.3, 129.1, 129.3, 130.4, 132.5, 132.5, 135.5, 135.9,
137.7, 140.2, 159.4, 161.6, 165.7, 168.4. HPLC>94.2%.
Example 23:
2-({[3-(3,5-difluoro-4-hydroxyphenyl)phenyl]methyl}sulfanyl)-6-oxo-4-(thi-
ophen-2-yl)-1,6-dihydropyrimidine-5-carbonitrile (Compound I-3)
##STR00294##
[0818] Step 1:
2-({[3-(3,5-difluoro-4-hydroxyphenyl)phenyl]methyl}sulfanyl)-6-oxo-4-(thi-
ophen-2-yl)-1,6-dihydropyrimidine-5-carbonitrile (22.1)
[0819] To a stirred solution of intermediate 1.4 (152 mg, 0.65
mmol) and intermediate 10.4 (250 mg, 0.77 mmol) in DMSO (6 mL) was
added DIPEA (0.13 mL, 0.72 mmol) and stirring was continued at rt
for 4h. The crude mixture was poured into water and the resulting
aqueous mixture was washed with EtOAc, acidified to pH 3, and
extracted with EtOAc (3.times.50 mL). The combined organic phases
were washed with brine and dried over Na.sub.2SO4 to afford
intermediate 22.1 (180 mg, 0.0.38 mmol) as a pale yellow powder
after flash chromatography purification eluting with
CH.sub.2Cl.sub.2/MeOH (4% for product). Yield 55%. .sup.1H NMR (400
MHz, DMSO-d.sub.6) .delta. 3.93 (s, 3H), 4.60 (s, 2H), 7.34-7.43
(m, 4H), 7.5 (d, J=4.3 Hz, 1H), 7.60 (d, J=7.5 Hz, 1H), 7.84 (s,
1H), 8.0 (d, J=4.9 Hz, 1H), 8.29 (d, J=4.9 Hz, 1H), 13.9 (brs,
1H).
Step 2:
2-({[3-(3,5-difluoro-4-hydroxyphenyl)phenyl]methyl}sulfanyl)-6-oxo-
-4-(thiophen-2-yl)-1,6-dihydropyrimidine-5-carbonitrile (Compound
I-3)
[0820] To a stirred suspension of intermediate 22.1 (170 mg, 0.36
mmol) in DCM (25 mL) was added a 1M solution of BBr.sub.3 in DCM
(0.72 mL, 0.72 mmol) and stirring was continued at reflux for 16h.
The reaction mixture was quenched by the addition of MeOH and the
volatiles were removed under reduced pressure. The crude product
was purified by flash chromatography eluting with DCM/MeOH (5% for
product). The title compound I-3 (90 mg, 0.2 mmol) was obtained as
a white solid after trituration with hot Et.sub.2O. Yield 42%.
.sup.1H NMR (400 MHz, DMSO) .delta. 4.59 (s, 3H), 7.31 (d, J=9.1
Hz, 2H), 7.36 (m, 1H), 7.39 (d, J=7.6 Hz, 1H), 7.45 (d, J=7.7 Hz,
1H), 7.56 (d, J=7.5 Hz, 1H), 7.81 (s, 1H), 8.07 (d, J=5 Hz, 1H),
8.3 (d, J=3.8 Hz, 1H), 10.34 (s, 1H). .sup.13C NMR (100 MHz,
DMSO-d.sub.6) .delta. 88.7, 110.3 (.sup.2J.sub.CF=15.1 Hz), 110.3
(.sup.2J.sub.CF=15.5 Hz), 125.8, 127.4, 128.5, 129.7. 130, 130.5,
132.1, 133.5 (.sup.3J.sub.CF=16 Hz), 133.7 (.sup.3J.sub.CF=16 Hz),
135.3, 138.1, 138.4, 139.7, 152.9 (.sup.1J.sub.CF=239.9 Hz), 153.01
(.sup.1J.sub.CF=240.1 MHz), 159.0, 161.3, 165.5. HPLC: 98.4%.
Example 24:
[3-(5-Cyano-4-methoxy-6-thiophen-2-yl-pyrimidin-2-ylsulfanylmethyl)-pheny-
l]-acetic acid (Compound I-4)
##STR00295##
[0822] To a stirred solution of intermediate 12.2 (80 mg, 0.19
mmol) and MeOH (0.04 mL, 0.95 mmol) in DMF (3 mL) was added
K.sub.2CO.sub.3 (60 mg, 0.43 mmol) and stirring was continued at rt
for 16h. The reaction mixture was poured into water and the
resulting aqueous mixture was extracted with EtOAc (3.times.20 mL).
The combined organic phase was washed with brine and dried over
Na.sub.2SO.sub.4. Flash chromatography purification of the crude
product (eluting with DCM/MeOH, 1.5% for product) afforded the
title compound I-4 (45 mg, 0.11 mmol) as a white solid. Yield 58%
o; .sup.1H NMR (400 MHz, DMSO-d.sub.6) .delta. 3.56 (s, 3H), 3.66
(s, 2H), 4.54 (s, 2H), 7.17 (d, J=7.2 Hz, 1H), 7.27 (t, J=7.6 Hz,
1H), 7.33-7.38 (m, 3H), 8.07 (d, J=4.8 Hz, 1H), 8.28 (d, J=3.6 Hz,
1H). .sup.13C NMR (100 MHz, DMSO-d.sub.6) .delta. 34.2, 40.3, 52.1,
88.5, 116.6, 127.8, 129, 129.1, 129.8, 130.3, 132, 135.1, 135.3,
137.1, 139.8, 159.1, 161.4, 165.5, 171.8. HPLC>97.1%.
Example 25:
[3-(4-Bromo-5-cyano-6-thiophen-2-yl-pyrimidin-2-ylsulfanylmethyl)-phenyl]-
-acetic acid (Compound I-5)
##STR00296##
[0824] To a stirred solution of intermediate 12.2 (30 mg, 0.051
mmol) in AcOH (3.0 mL) was added HBr (36% solution in AcOH, 0.17
mL, 1.029 mmol) and the resulting mixture was stirred at 60.degree.
C. for 72 h. The reaction mixture was then diluted with DCM (10
mL), washed with H.sub.2O (3.times.10 mL), brine (10 mL), dried
over Na.sub.2SO.sub.4 and concentrated under reduced pressure. The
crude product was purified by flash chromatography (DCM/MeOH/AcOH,
from 99:1:0.1 92:8:0.1) to afford the title compound I-5 (17 mg,
0.038 mmol) as a yellow solid. Yield 75%. MS/MS ESI (+): 447.8,
401.9, 338.3. .sup.1H-NMR (CDCl.sub.3, 400 MHz) .delta.: 3.63 (s,
2H), 4.45 (s, 2H), 7.21 (m, 1H), 7.31 (m, 2H), 7.39 (m, 2H), 7.70
(brs, 1H), 8.45 (brs, 1H). .sup.13C-NMR (CDCl.sub.3, 100 MHz)
.delta.: 29.3, 40.7, 100.1, 116.0, 127.9, 128.7, 128.9, 129.4,
130.1, 133.1, 133.6, 134.6, 136.6, 138.7, 155.8, 159.7, 174.3,
177.1. HPLC>95%.
Example 26:
[3-(5-Cyano-4-cyclopropylamino-6-thiophen-2-yl-pyrimidin-2-ylsulfanylmeth-
yl)-phenyl]-acetic acid (Compound I-6)
##STR00297##
[0826] To a stirred solution of intermediate 12.2 (200 mg, 0.49
mmol) in DMF (3 mL) was added cyclopropylamine (0.037 mL, 0.55
mmol) and stirring was continued at r.t. for 16 h. The reaction
mixture was quenched with brine, poured into water, and the
resulting aqueous mixture was extracted with EtOAc (3.times.20 mL).
The combined organic phases were washed with brine and dried over
Na.sub.2SO.sub.4. The title compound I-6 (80 mg, 0.2 mmol) was
obtained as a white solid after shredding with Et.sub.2O. Yield
39%. .sup.1H NMR (400 MHz, DMSO-d.sub.6) .delta. 0.73 (m, 4H), 2.95
(m, 1H), 3.53 (s, 2H), 4.45 (s, 2H), 7.13 (d, J=7.4 Hz, 1H),
7.23-7.29 (m, 2H), 7.33-7.35 (m, 2H), 7.93 (d, J=4.9 Hz, 1H), 8.18
(d, J=3.2 Hz, 1H), 8.21 (s, 1H), 12.3 (s, 1H). .sup.13C NMR (100
MHz, DMSO-d.sub.6) .delta. 6.6, 6.6, 25, 34.3, 40.8, 79.7, 116.7,
127.3, 128.5, 128.6, 129.2, 130.1, 130.7, 133.2, 135.4, 138.3,
140.1, 158.7, 162.8, 172.8, 172.8. HPLC>99.3%.
Example 27:
[3-(4-Amino-5-cyano-6-thiophen-2-yl-pyrimidin-2-ylsulfanylmethyl)-phenyl]-
-acetic acid (Compound I-7)
##STR00298##
[0828]
[3-(4-Cloro-5-cyano-6-thiophen-2-yl-pyrimidin-2-ylsulfanylmethyl)-p-
henyl]-acetic acid (12.2) (100 mg, 0.248 mmol) was dissolved in
NH.sub.3 0.4 M in THE (18 mL, 7.466 mmol) and the resulting
opalescent solution was stirred at room temperature for 72 hours.
Then the mixture was poured in AcOEt (15 ml), washed with HCl 3 M
(5 mL), aq. NaHCO.sub.3 ss (10 mL), brine (10 mL) dried over
Na.sub.2SO.sub.4 and concentrated under reduced pressure. The crude
was purified by flash chromatography (CH.sub.2Cl.sub.2/MeOH/AcOH,
from 99:1:0.1 90:10:0.1) to afford the title compound I-7 (86 mg,
0.22 mmol) as a white solid. Yield 94%; MS/MS ESI (+): 382.9.
.sup.1H NMR (400 MHz, DMSO-d.sub.6) .delta. 3.53 (s, 2H), 4.39 (s,
2H), 7.13 (d, J=7.3 Hz, 1H), 7.24 (t, J=7.1 Hz, 1H), 7.29 (m, 1H),
7.35 (m, 2H), 7.8 (brs, 1H), 7.94 (d, J=4.3 Hz, 1H), 8.20 (m, 1H),
12.32 (brs, 1H). .sup.13C NMR (100 MHz, DMSO-d.sub.6) .delta. 34.3,
40.9, 78.9, 116.9, 127.6, 128.6, 128.7, 129.3, 130.4, 130.8, 133.4,
135.5, 138.3, 140.4, 159.1, 163.7, 173. HPLC>97.9%.
Example 28:
[3-(5-Cyano-6-thiophen-2-yl-pyrimidin-2-ylsulfanylmethyl)-phenyl]-acetic
acid (Compound I-8)
##STR00299##
[0830] Et.sub.3N (0.15 mL, 1.119 mmol) was added to a stirred
solution of intermediate 12.2 (150 mg, 0.373 mmol) in THE (3.7 mL).
The resulting solution was continuously hydrogenated for 12 h using
the Thales Nano H-Cub Hydrogenator (Cartridge: Pd/C 10%, H.sub.2
Pressure: 8 bar, temperature: 40.degree. C., transporting solvent:
THF, flowrate: 1.0 mL/min). The resulting reaction mixture (about 5
mL) was diluted with EtOAc (15 mL), washed with 3M HCl (5 mL) and
brine (10 mL), dried over Na.sub.2SO.sub.4, filtered, and
concentrated under reduced pressure. The crude product was purified
by reverse-phase flash chromatography (column: RP-18, eluting with
H.sub.2O/MeOH 80/20 to 10/90) to give the title compound I-8 as a
white powder. Yield 34%. MS/MS ESI (+): 368.1. .sup.1H-NMR
(DMSO-d.sub.6, 400 MHz) .delta.: 3.60 (s, 2H), 4.45 (s, 2H), 7.15
(m, 1H), 7.25 (m, 1H), 7.36 (m, 4H), 8.06 (br-s, 1H), 8.30 (ps-s,
1H), 9.03 (s, 1H).
Example 29:
[3-(5-Cyano-4-methylamino-6-thiophen-2-yl-pyrimidin-2-ylsulfanylmethyl)-p-
henyl]-acetic acid (Compound I-9)
##STR00300##
[0832] To a stirred solution of intermediate 12.2 (100 mg, 0.25
mmol) in DMF (3 mL) was added a 33% solution of MeNH.sub.2 (0.03
mL, 0.27 mmol) in ethanol and stirring was continued at rt for 16h.
The reaction mixture was quenched with brine, poured into water,
acidified to pH 6 by the addition of a 3M HCl solution, and then
extracted with EtOAc (3.times.20 mL). The combined organic phase
was washed with brine and dried over Na.sub.2SO.sub.4 to afford the
title compound I-9 (80 mg, 0.2 mmol) as a white solid. Yield 80%.
.sup.1H NMR (400 MHz, DMSO-d.sub.6) .delta. 2.93 (d, J=4.5 Hz, 3H),
3.53 (s, 2H), 2.92 (s, 2H), 7.13 (d, J=7.6 Hz, 1H), 7.24 (d, J=7.5
Hz, 1H), 7.27-7.29 (m, 1H), 7.33 (m, 2H), 7.94 (d, J=5.1 Hz, 1H),
8.1 (q, J=4.5 Hz, 1H), 8.18 (d, J=3.8 Hz, 1H), 12.33 (s, 1H).
.sup.13C NMR (100 MHz, DMSO-d.sub.6) .delta. 28.6, 34.5, 40.9,
79.6, 116.9, 127.4, 128.6, 128.7, 129.3, 130.2, 130.7, 133.3,
135.5, 138.3, 140.3, 158.5, 161.9, 172.9, 173.1. HPLC>98.1%.
Example 30:
3-(5-Cyano-4-methylamino-6-thiophen-2-yl-pyrimidin-2-ylsulfanylmethyl)-be-
nzoic acid (Compound I-10)
##STR00301##
[0834] To a stirred solution of intermediate 13.3 (90 mg, 0.23
mmol) in DMF (3 mL) was added a 33% solution of MeNH.sub.2 (0.03
mL, 0.25 mmol) in ethanol and stirring was continued at rt for 16h.
The reaction mixture was quenched with brine, poured into water,
acidified to pH 6 by the addition of a 3M HCl solution, and then
extracted with EtOAc (3.times.20 mL). The combined organic phase
was washed with brine and dried over Na.sub.2SO.sub.4. The crude
product was purified by flash chromatography eluting with DCM/MeOH
(4% for product) to afford the title compound I-10 (45 mg, 0.12
mmol) as a white solid. Yield 51%. .sup.1H NMR (400 MHz,
DMSO-d.sub.6) .delta. 2.94 (d, J=4.4 Hz, 3H), 4.49 (s, 2H), 7.28
(t, J=4.4 Hz, 1H), 7.44 (t, J=7.7 Hz, 1H), 7.70 (d, J=7.5 Hz, 1H),
7.81 (d, J=7.7 Hz, 1H), 7.92 (d, J=5. Hz, 1H), 8.04 (q, J=4.5 Hz,
1H), 8.06 (s, 1H)), 8.18 (d, J=3.8 Hz, 1H), 12.92 (s, 1H). .sup.13C
NMR (100 MHz, DMSO-d.sub.6) .delta. 28.6, 34.2, 79.8, 116.8, 128.3,
129.1, 129.2, 130, 130.7, 131.3, 133.3, 133.5, 139.3, 140.2, 158.5,
161.9, 167.4, 172.9. HPLC>95.1%.
Example 31:
2-(3-Cyano-benzylsulfanyl)-6-oxo-4-thiophen-2-yl-1,6-dihydro-pyrimidine-5-
-carbonitrile (Compound I-11)
##STR00302##
[0836] To a stirred solution of intermediate 1.4 (200 mg, 0.85
mmol) and K.sub.2CO.sub.3 (133 mg, 0.93 mmoL) in acetone (20 mL)
was added intermediate 14.2 (133 mg, 0.93 mmol) and stirring was
continued at rt for 16h. The solvent was then removed under reduced
pressure. The resulting mixture was poured into water, acidified to
pH 6 by the addition of a 3M HCl solution, and then extracted with
EtOAc (3.times.20 mL). The combined organic phase was washed with
brine and dried over Na.sub.2SO.sub.4. The crude product was
purified by flash chromatography eluting with DCM/MeOH to provide
the title compound I-11 (50 mg, 0.17 mmol) as a white solid. Yield
17%. .sup.1H NMR (400 MHz, DMSO-d.sub.6) .delta. 4.60 (s, 2H), 7.35
(d, J=4.8 Hz, 1H), 7.54 (t, J=7.8 Hz, 1H), 7.74 (d, J=7.7 Hz, 1H),
7.83 (d, J=7.9 Hz, 1H), 7.96 (s, 1H), 8.1 (d, J=5.02 Hz, 1H), 8.27
(d, J=3.9 Hz, 1H), 13.80 (brs, 1H). .sup.13C NMR (100 MHz,
DMSO-d.sub.6) .delta. 33.2, 88.8, 111.8, 116.5, 118.9, 130, 130.2,
131.6, 132.1, 132.8, 134.1, 135.4, 139.3, 139.6, 159, 161.2, 165.1.
HPLC>99.1%.
Example 32:
2-[3-(2-Hydroxy-ethyl)-benzylsulfanyl]-6-oxo-4-thiophen-2-yl-1,6-dihydrop-
yrimidine-5-carbonitrile (Compound I-12)
##STR00303##
[0838] To a stirred solution of intermediate 1.4 (200 mg, 0.85
mmol) and DIPEA (0.16 mL, 0.93 mmoL) in acetone (15 mL) was added
intermediate 15.1 (201 mg, 0.93 mmol) and stirring was continued at
rt for 16h. The solvent was then removed under reduced pressure.
The resulting mixture was poured into water, acidified to pH 6 by
the addition of a 3M HCl solution, and then extracted with EtOAc
(3.times.20 mL). The combined organic phase was washed with brine
and dried over Na.sub.2SO.sub.4. The crude product was purified by
flash chromatography eluting with DCM/MeOH to provide the title
compound I-12 (120 mg, 0.32 mmol) as a white solid. Yield 38%.
.sup.1H NMR (400 MHz, DMSO-d.sub.6) .delta. 2.47, (t, J=8.35 Hz,
2H), 2.67 (t, J=7.01 Hz, 2H), 4.48 (s, 2H), 4.51 (brs, 1H), 7.11
(d, J=7.5 Hz, 1H), 7.22 (t, J=7.5 Hz, 1H), 7.29 (d, J=7.7 Hz, 1H),
7.32-7.36 (m, 2H), 8.06 (d, J=4.9 Hz, 1H), 8.27 (d, J=3.9 Hz, 1H),
13.80 (brs, 1H). .sup.13C NMR (100 MHz, DMSO-d.sub.6) .delta. 34.3,
62.3, 62.3, 88.3, 116.9, 126.8, 128.5, 128.8, 129.8, 129.9, 131.8,
135.1, 136.9, 139.9, 140.3, 159, 162, 165.9. HPLC>96.1%.
Example 33:
2-(3-Cyanomethyl-benzylsulfanyl)-6-oxo-4-thiophen-2-yl-1,6-dihydro-pyrimi-
dine-5-carbonitrile (Compound I-13)
##STR00304##
[0840] To a stirred solution of intermediate 1.4 (200 mg, 0.85
mmol) and DIPEA (0.2 mL, 0.94 mmoL) in acetone (20 mL) was added
intermediate 16.2 (196 mg, 0.94 mmol) and stirring was continued at
rt for 16h. The solvent was then removed under reduced pressure.
The resulting mixture was poured into water, acidified to pH 6 by
the addition of a 3M HCl solution, and then extracted with EtOAc
(3.times.20 mL). The combined organic phase was washed with brine
and dried over Na.sub.2SO.sub.4. The crude product was purified by
flash chromatography eluting with DCM/MeOH (2.5% for product) to
provide the title compound I-13 (300 mg, 0.82 mmol) as a yellow
solid. Yield 96%. .sup.1H NMR (400 MHz, DMSO-d.sub.6) .delta. 4.01
(s, 2H), 4.52 (s, 2H), 7.24 (d, J=7.49 Hz, 1H), 7.31-7.36 (m, 2H),
7.43-7.45 (m, 2H), 8.0 (d, J=5 Hz, 1H), 8.23 (d, J=3.8 Hz, 1H),
13.80 (brs, 1H). .sup.13C NMR (100 MHz, DMSO-d.sub.6) .delta. 22.6,
33.9, 87.9, 117.5, 119.5, 127.5, 128.5, 128.9, 129.6, 129.6, 131.3,
131.9, 134.5, 138.6, 140.3, 159.1, 164.1, 166.7. HPLC>97.7%.
Example 34:
2-[3-(2-Methyl-2H-tetrazol-5-yl)-benzylsulfanyl]-6-oxo-4-thiophen-2-yl-1,-
6-dihydro-pyrimidine-5-carbonitrile (Compound I-14)
##STR00305##
[0842] To a stirred solution of intermediate 1.4 (200 mg, 0.85
mmol) and DIPEA (0.17 mL, 0.93 mmoL) in acetone (15 mL) was added
intermediate 17.3 (236 mg, 0.93 mmol) and stirring was continued at
rt for 16h. The solvent was then removed under reduced pressure.
The resulting solid was collected and dried under reduced pressure
to give the title compound I-14 (200 mg, 0.49 mmol) as a yellowish
solid. Yield 58%. .sup.1H NMR (400 MHz, DMSO-d.sub.6) .delta. 4.40
(s, 3H), 4.66 (s, 2H), 7.35 (m, 1H), 7.51 (t, J=7.5 Hz, 1H), 7.66
(d, J=6.9 Hz, 1H), 7.94 (d, J=7.2 Hz, 1H), 8.08 (d, J=4.1 Hz, 1H),
8.21 (s, 1H), 8.28 (s, 1H), 13.80 (s, 1H). .sup.13C NMR (100 MHz,
DMSO-d.sub.6) .delta. 33.9, 40.5, 88.7, 116.5, 125.7, 127.2, 127.5,
129.9, 130, 131.3, 132.1, 135.4, 138.6, 139.7, 159.1, 161.1, 164.2,
165.2. HPLC>99.3%.
Example 35:
[3-(5-Cyano-4-morpholin-4-yl-6-thiophen-2-yl-pyrimidin-2-ylsulfanylmethyl-
)-phenyl]-acetic acid (Compound I-15)
##STR00306##
[0844] To a stirred suspension of intermediate 12.2 (100 mg, 0.25
mmol) in CH.sub.3CN (10 mL) was added morpholine (0.023 mL, 0.27
mmol) and stirring was continued at rt for 16h.
[0845] The solvent was then removed under reduced pressure. The
crude product was taken up in water, and the resulting aqueous
mixture was extracted with EtOAc (3.times.20 mL). The combined
organic phase was washed with brine and dried over Na.sub.2SO.sub.4
to provide the title compound I-15 (80 mg, 0.18 mmol) as a white
solid. Yield 71%. .sup.1H NMR (400 MHz, CDCl.sub.3) .delta. 3.60
(s, 2H), 3.79 (m, 4H), 3.93 (m, 4H), 4.41 (s, 2H), 7.17-7.20 (m,
2H), 7.27-7.36 (m, 2H), 7.37 (d, J=8.23 Hz, 2H), 7.61 (d, J=5.1 Hz,
1H), 8.32 (d, J=3.5 Hz, 1H). .sup.13C NMR (100 MHz, CDCl.sub.3)
.delta. 35.1, 40.6, 47.7, 47.7, 66.5, 66.5, 80.6, 118.4, 127.7,
128.3, 128.6, 128.8, 129.7, 131.7, 132.4, 133.5, 137.6, 139.9,
161.6, 162.9, 172.6, 176.3. HPLC>98.1%.
Example 36:
[3-(5-Cyano-4-piperazin-1-yl-6-thiophen-2-yl-pyrimidin-2-ylsulfanylmethyl-
)-phenyl]-acetic acid (Compound I-16)
##STR00307##
[0846] Step 1.
4-[2-(3-Carboxymethyl-benzylsulfanyl)-5-cyano-6-thiophen-2-yl-pyrimidin-4-
-yl]-piperazine-1-carboxylic acid tert-butyl ester (36.1)
[0847] To a stirred suspension of intermediate 12.2 (250 mg, 0.62
mmol) and K.sub.2CO.sub.3 (128 mg, 0.93 mmol) in DMF (4 mL) was
added 1-boc-piperazine (127 mg, 0.68 mmol) and stirring was
continued at rt for 16h. The solvent was then removed under reduced
pressure. The resulting mixture was taken up in water, and the
aqeuous mixture was extracted with EtOAc (3.times.20 mL). The
combined organic phase was washed with brine and dried over
Na.sub.2SO4. The crude product was purified by flash chromatography
eluting with DCM/MeOH (4% for product) to provide the title
intermediate 35.1 (60 mg, 0.11 mmol) as a yellowish solid. Yield
18%.
Step 2.
[3-(5-Cyano-4-piperazin-1-yl-6-thiophen-2-yl-pyrimidin-2-ylsulfany-
lmethyl)-phenyl]-acetic acid (I-16)
[0848] To a stirred solution of intermediate 35.1 (65 mg, 0.12
mmol) in DCM (15 mL) was added TFA (0.28 mL, 3.6 mmol) and stirring
was continued at rt for 16h. The solvent was then removed under
reduced pressure. The crude mixture was taken up in water, and the
resulting aqueous mixture was extracted with EtOAc (3.times.20 mL).
The combined organic phase was washed with brine and dried over
Na.sub.2SO4. The title compound I-16 (20 mg, 0.044 mmol) was
obtained as a white solid after shredding with hot Et.sub.2O. Yield
37%. .sup.1H NMR (400 MHz, DMSO-d.sub.6) .delta. 2.83 (m, 4H), 3.51
(s, 2H), 3.81 (m, 4H), 4.39 (s, 3H), 7.13 (d, J=7.03 Hz, 1H),
7.24-7.33 (m, 4H), 7.95 (d, J=4.4 Hz, 1H), 8.20 (d, J=2.8 Hz, 1H).
.sup.13C NMR (100 MHz, DMSO-d.sub.6) .delta. 34.6, 41.2, 45.5,
45.5, 48.4, 48.4, 118.7, 127.2, 128.6, 128.7, 129.2, 130.1, 131.8,
133.7, 135.8, 138.1, 140.1, 161.5, 162.4, 172, 173.1.
HPLC>90.9%.
Example 37.
[3-(5-Cyano-1-methyl-4-oxo-6-thiophen-2-yl-1,4-dihydro-pyrimidin-2-ylsulf-
anylmethyl)-phenyl]-acetic acid ethyl ester (Compound I-17)
##STR00308##
[0850] To a stirred suspension of compound I-1 (300 mg, 0.73 mmol)
and K.sub.2CO.sub.3 (151 mg, 1.09 mmol) in DMF (15 mL) was added
Mel (0.047 mL, 0.77 mmol) dropwise and stirring was continued at rt
for 16h. The resulting mixture was poured into water, and then
extracted with EtOAc (3.times.20 mL). The combined organic phase
was washed with brine and dried over Na.sub.2SO4 to provide the
title compound I-17 (298 mg, 0.7 mmol) as a yellowish solid. Yield
96%. .sup.1H NMR (400 MHz, DMSO-d.sub.6) .delta. 0.14 (t, J=7.12
Hz, 3H), 3.41 (s, 3H), 3.62 (s, 2H), 4.03 (q, J=7.1 Hz, 2H), 4.64
(s, 2H), 7.19 (d, J=7.6 Hz, 1H), 7.27-7.31 (m, 1H), 7.35 (t, J=4.1
Hz, 1H), 7.40 (m, 2H), 8.08 (d, J=4.9 Hz, 1H), 8.28 (d, J=3.8 Hz,
1H). .sup.13C NMR (100 MHz, DMSO-d.sub.6) .delta. 14.4, 31.1, 36.1,
40.5, 60.6, 87.5, 116.5, 127.9, 129.1, 129.2, 130, 130.4, 132.1,
135.3, 135.4, 136.2, 139.5, 157.1, 160.1, 166.4, 171.3.
HPLC>95.1%.
Example 38.
3-(5-Cyano-6-oxo-4-thiophen-2-yl-1,6-dihydro-pyrimidin-2-ylsulfanylmethyl-
)-benzenesulfonamide (Compound I-18)
##STR00309##
[0852] To a stirred solution of intermediate 1.4 (107 mg, 0.45
mmol) and DIPEA (0.08 mL, 0.49 mmoL) in acetone (15 mL) was added
intermediate 19.5 (125 mg, 0.49 mmol) and stirring was continued at
rt for 16h. The solvent was then removed under reduced pressure.
The crude mixture was taken up in water, and then extracted with
EtOAc (3.times.20 mL). The combined organic phase was washed with
brine and dried over Na.sub.2SO.sub.4. The title compound I-18 (50
mg, 0.12 mmol) was obtained as a yellowish solid after trituration
with hot Et.sub.2O. Yield 28%. .sup.1H NMR (400 MHz, DMSO-d.sub.6)
.delta. 4.65 (s, 2H), 7.34 (t, J=4.6 Hz, 1H), 7.40 (s, 2H), 7.52
(t, J=7.6 Hz, 1H), 7.72 (t, J=6.1 Hz, 2H), 7.94 (s, 1H), 8.06 (d,
J=4.8 Hz, 1H), 8.27 (d, J=3.6 Hz, 1H), 13.8 (s, 1H). .sup.13C NMR
(100 MHz, DMSO-d.sub.6) .delta. 33.7, 88.6, 116.6, 125.2, 126.1,
129.7, 129.9, 132, 132.5, 135.4, 138.4, 139.7, 144.8, 159.1, 161.4,
165.3. HPLC>95.1%.
Example 39:
[3-(3-Cyano-6-oxo-4-phenyl-1,6-dihydro-pyridin-2-ylsulfanylmethyl)-phenyl-
]-acetic acid (Compound I-19)
##STR00310##
[0854] To a stirred suspension of intermediate 3.3 (100 mg, 0.44
mmol) and DIPEA (0.09 mL, 0.53 mmol) in acetone (15 mL) was added
intermediate 11.2 (94 mg, 0.44 mmol). Stirring was continued
overnight at rt. The mixture was diluted with crushed ice and
water. pH was adjusted to 5 by the addition of AcOH. The
precipitate was collected, washed with cold water and dried under
vacuo. Compound I-19 (60 mg, 0.16 mmol) was obtained as a brownish
powder. Yield 37%. .sup.1H NMR (400 MHz, DMSO-d.sub.6) .delta. 3.55
(s, 2H), 4.5 (s, 2H), 6.51 (s, 1H), 7.16 (d, J=7.4 Hz, 1H), 7.27
(t, J=7.7 Hz, 1H), 7.37 (m, 2H), 7.51-7.53 (m, 3H), 7.55-7.56 (m,
2H), 12.17 (brs, 2H); .sup.13C NMR (100 MHz, DMSO-d.sub.6) .delta.
33.5, 40.6, 95.9, 108, 116.2, 127.6, 128.3, 128.3, 128.5, 128.5,
128.9, 128.9, 130, 130.3, 135.4, 135.9, 137.5, 155.9, 162.1, 164.9,
172.7; HPLC: 96.88%.
Example 40:
[3-(3-Cyano-6-oxo-4-thiophen-2-yl-1,6-dihydro-pyridin-2-ylsulfanylmethyl)-
-phenyl]-acetic acid (Compound I-20)
##STR00311##
[0856] To a stirred suspension of intermediate 5.2 (153 mg, 0.56
mmol) and DIPEA (0.12 mL, 0.67 mmol) in DMSO/acetone (15/4 mL) was
added intermediate 11.2 (121 mg, 0.56 mmol). Stirring was continued
overnight at room temperature. The mixture was diluted with crushed
ice and water. The pH was adjusted to 5 by the addition of AcOH.
The precipitate was collected washed with cold water and dried
under vacuo. Compound I-20 (90 mg, 0.22 mmol) was obtained as a
brownish powder. Yield 42%. .sup.1H NMR (400 MHz, DMSO-d.sub.6)
.delta. 3.55 (s, 2H), 4.51 (s, 2H), 6.62 (s, 1H), 7.15 (d, J=7.4
Hz, 1H), 7.24-7.28 (m, 2H), 7.35 (d, J=6.4 Hz, 2H), 7.75 (d, J=3.5
Hz, 1H), 7.85 (d, J=4.9 Hz, 1H), 12.1 (brs, 1H); .sup.13C NMR (100
MHz, DMSO-d.sub.6) .delta. 33.6, 40.6, 93.8, 104.5, 116.5, 127.6,
128.6, 128.6, 128.7, 128.7, 129.5, 130.3, 130.3, 135.4, 136.6,
137.4, 147.4, 165.1, 172.7; HPLC: 96.5%.
Example 41:
[3-(3,5-Dicyano-6-oxo-4-thiophen-2-yl-1,6-dihydro-pyridin-2-ylsulfanylmet-
hyl)-phenyl]-acetic acid (Compound I-21)
##STR00312##
[0858] To a stirred solution of intermediate 6.2 (200 mg, 0.77
mmol) and DIPEA (0.16 mL, 0.92 mmol) in acetone (15 mL) was added
intermediate 11.2 (165 mg, 0.77 mmol). Stirring was continued
overnight at room temperature. The mixture was diluted with crushed
ice and water. The pH was adjusted to 5 by the addition of AcOH.
The precipitate was collected washed with cold water and dried
under vacuo. Compound I-21 (110 mg, 0.27 mmol) was obtained as a
brownish powder. Yield 35%. .sup.1H NMR (400 MHz, DMSO-d.sub.6)
.delta. 3.56 (s, 2H), 4.48 (s, 2H), 7.63 (d, J=7.6 Hz, 1H),
7.24-7.28 (m, 2H), 7.38-7.40 (m, 2H), 7.54 (dd, J=1.1 Hz, J=3.6 Hz,
1H), 7.93 (dd, J=1.1 Hz, J=5 Hz, 1H), 8.12 (brs, 1H), 12.29 (brs,
1H); .sup.13C NMR (100 MHz, DMSO-d.sub.6) .delta. 33.3, 40.6, 85.8,
93.1, 115.5, 127.8, 128, 128, 128.6, 130.5, 130.9, 131.4, 131.4,
132.9, 135.3, 137.4, 150.8, 159.8, 166.9, 172.8; HPLC: 97.5%.
Example 42:
2-Oxo-6-[3-(1H-tetrazol-5-yl)-benzylsulfanyl]-4-thiophen-2-yl-1,2-dihydro-
-pyridine-3,5-dicarbonitrile (Compound I-22)
##STR00313##
[0860] To a stirred solution of intermediate 6.2 (150 mg, 0.57
mmol) and DIPEA (0.18 mL, 0.68 mmol) in acetone (15 mL) was added
intermediate 18.1 (138 mg, 0.57 mmol). Stirring was continued
overnight at room temperature. The mixture was diluted with crushed
ice and water. pH was adjusted to 5 by the addition of AcOH. The
precipitate was collected washed with cold water and dried under
vacuo. Compound I-22 (90 mg, 0.27 mmol) was obtained as a yellowish
powder. Yield 38%. .sup.1H NMR (400 MHz, DMSO-d.sub.6) .delta. 4.6
(s, 2H), 7.26 (dd, J=5.0 Hz, J=3.6 Hz, 1H), 7.54-7.56 (m, 2H), 7.76
(d, J=7.7 Hz, 1H), 7.90-7.94 (m, 2H), 8.1 (s, 1H); .sup.13C NMR
(100 MHz, DMSO-d.sub.6) .delta. 33.2, 86.2, 93.4, 115.7, 124.9,
126.2, 128.1, 128.2, 129.8, 131.2, 131.6, 131.6, 132.5, 133, 139.5,
151.1, 155.8, 160.1, 166.8; HPLC: 96.7%
Example 43:
[3-(4-Benzyl-3-cyano-6-oxo-1,6-dihydro-pyridin-2-ylsulfanylmethyl)-phenyl-
]-acetic acid (Compound I-23)
##STR00314##
[0862] To a stirred solution of intermediate 20.2 (154 mg, 0.63
mmol) and DIPEA (0.12 mL, 0.7 mmol) in acetone (15 mL) was added
intermediate 11.2 (150 mg, 0.7 mmol). Stirring was continued
overnight at room temperature. The mixture was diluted with crushed
ice and water. pH was adjusted to 5 by the addition of AcOH. The
precipitate was collected washed with cold water and dried under
vacuo. Compound I-23 (100 mg, 0.25 mmol) was obtained as a
yellowish powder. Yield 40%). .sup.1H NMR (400 MHz, DMSO-d.sub.6)
.delta. 3.37 (s, 2H), 3.98 (s, 2H), 4.48 (s, 2H), 6.34 (s, 1H),
7.13-7.32 (m, 9H), 12.1 (brs, 1H); HPLC: 98.5%.
Example 44:
(5-Cyano-6-oxo-4-thiophen-2-yl-1,6-dihydro-pyrimidin-2-ylsulfanyl)-acetic
acid (Compound I-24)
##STR00315##
[0864] To a stirred solution of intermediate 1.4 (200 mg, 0.85
mmol) and DIPEA (0.18 mL, 1.02 mmol) in acetone/DMSO (20:2 mL) was
added chloroacetic acid (80 mg, 0.85 mmol). Stirring was continued
overnight at room temperature. Additional 0.3 equivalents of DIPEA
and of chloroacetic acid were then added to complete the reaction.
The mixture was diluted with crushed ice and water. pH was adjusted
to 3 by the addition of 3N HCl. The precipitate was collected and
purified by reverse flash chromatography, eluting with
H.sub.2O/MeOH from 10 to 80%. Compound I-24 (210 mg, 0.71 mmol) was
obtained as a yellowish powder. Yield 83%. .sup.1H NMR (400 MHz,
DMSO-d.sub.6) .delta. 4.0 (s, 2H), 7.33 (t, J=4.7 Hz, 1H), 8.1 (d,
J=4.9 Hz, 1H), 8.25 (d, J=3.8 Hz, 1H), 12.8 (brs, 1H); .sup.13C NMR
(100 MHz, DMSO-d.sub.6) .delta. 33.4, 88.5, 116.5, 129.8, 132.3,
135.6, 139.5, 159, 161.1, 165.2, 169.3; HPLC: 99.6%.
Example 45:
6-Oxo-2-(1H-tetrazol-5-ylmethylsulfanyl)-4-thiophen-2-yl-1,6-dihydro-pyri-
midine-5-carbonitrile (Compound I-25)
##STR00316##
[0866] To a stirred solution of intermediate 1.4 (200 mg, 0.85
mmol) and DIPEA (0.18 mL, 1.02 mmol) in acetone/DMSO (20:2 mL) was
added intermediate 5-chloromethyl-1H-tetrazole (101 mg, 0.85 mmol).
Stirring was continued overnight at room temperature. Additional
0.3 equivalents of DIPEA and of 5-chloromethyl-1H-tetrazole were
then added to complete the reaction. The mixture was diluted with
crushed ice and water. pH was adjusted to 3 by the addition of 3N
HCl. The precipitate was collected and purified by reverse flash
chromatography, eluting with H.sub.2O/MeOH from 10 to 80%. Compound
I-25 (120 mg, 0.37 mmol) was obtained as a yellowish powder. Yield
44%. .sup.1H NMR (400 MHz, DMSO-d.sub.6) .delta. 4.82 (s, 2H), 7.31
(t, J=4.2 Hz, 1H), 8.0 (d, J=4.9 Hz, 1H), 8.22 (d, J=3.8 Hz, 1H);
.sup.13C NMR (100 MHz, DMSO-d.sub.6) .delta. 23.6, 88.7, 116.4,
129.9, 132.2, 135.6, 139.4, 157.1, 159.0, 161.4, 164.4; HPLC:
94.6%.
Example 46:
2-(1H-Tetrazol-5-ylmethylsulfanyl)-6-trifluoromethyl-3H-pyrimidin-4-one
(Compound I-26)
##STR00317##
[0868] To a stirred solution of intermediate 2.1 (100 mg, 0.56
mmol) and DIPEA (0.13 mL, 0.73 mmol) in acetone (5 mL) was added
5-chloromethyl-1H-tetrazole (87 mg, 0.73 mmol). Stirring was
continued overnight at room temperature. The mixture was diluted
with crushed ice and water. pH was adjusted to 3 by the addition of
3N HCl. The aqueous phase was extracted with EtOAc (3.times.20 mL).
The combined organic phase was washed with brine and dried over
Na.sub.2SO.sub.4. Compound I-26 (60 mg, 0.19 mmol) was obtained as
a white powder. Yield 35%. .sup.1H NMR (400 MHz, DMSO-d.sub.6)
.delta. 4.69 (s, 2H), 6.67 (s, 1H), 15.1 (brs, 1H); .sup.13C NMR
(100 MHz, DMSO-d.sub.6) .delta. 23.1, 107.7, 120.6 (q, J.sub.CF=2.7
Hz), 152, 154.3, 163.9, 164.8; HPLC: 97.9%
Example 47:
(6-Oxo-4-trifluoromethyl-1,6-dihydro-pyrimidin-2-ylsulfanyl)-acetic
acid (Compound I-27)
##STR00318##
[0870] To a stirred solution of intermediate 2.1 (200 mg, 0.85
mmol) and DIPEA (0.16 mL, 0.94 mmol) in DMSO (5 mL) was added
chloroacetic acid (89 mg, 0.94 mmol). Stirring was continued
overnight at rt. The mixture was diluted with crushed ice and
water. pH was adjusted to 3 by the addition of 3N HCl. The aqueous
phase was extracted with EtOAc (3.times.20 mL). The combined
organic phase was washed with brine and dried over
Na.sub.2SO.sub.4. The crude of reaction was purified by reverse
flash chromatography eluting with H.sub.2O/MeOH from 5 to 65% for
product. Compound I-27 (125 mg, 0.49 mmol) was obtained as a white
powder. Yield 58%. .sup.1H NMR (400 MHz, DMSO-d.sub.6) .delta. 3.96
(s, 2H), 6.63 (s, 1H); .sup.13C NMR (100 MHz, DMSO-d.sub.6) .delta.
33.1, 108, 120.6 (q, J.sub.CF=2.7 Hz), 163.1, 165.4, 169.5; HPLC:
95.9%.
Example 48:
[3-(6-Oxo-4-trifluoromethyl-1,6-dihydro-pyrimidin-2-ylsulfanylmethyl)-phe-
nyl]-acetic acid (Compound I-28)
##STR00319##
[0872] To a stirred solution of intermediate 2.1 (150 mg, 0.64
mmol) and DIPEA (0.12 mL, 0.71 mmol) in DMSO (5 mL) was added
intermediate 11.2 (152 mg, 0.71 mmol). Stirring was continued
overnight at rt. The mixture was diluted with crushed ice and
water. pH was adjusted to 3 by the addition of 3N HCl. The aqueous
phase was extracted with EtOAc (3.times.20 mL). The combined
organic phase was washed with brine and dried over
Na.sub.2SO.sub.4. The crude of reaction was purified by reverse
flash chromatography eluting with H.sub.2O/MeOH from 5 to 80% for
product. Compound I-28 (100 mg, 0.29 mmol) was obtained as a white
powder. Yield 45%. .sup.1H NMR (400 MHz, DMSO-d.sub.6) .delta. 3.54
(s, 2H), 4.52 (s, 2H), 6.87 (s, 1H), 7.16 (d, J=7.3 Hz, 1H), 7.26
(t, J=7.4 Hz, 1H), 7.33-7.35 (m, 2H), 12.18 (brs, 1H); HPLC:
98.1%.
Example 49:
2-[3-(1H-Tetrazol-5-yl)-benzylsulfanyl]-6-trifluoromethyl-3H-pyrimidin-4--
one (Compound I-29)
##STR00320##
[0874] To a stirred solution of intermediate 2.1 (150 mg, 0.64
mmol) and DIPEA (0.12 mL, 0.71 mmol) in DMSO (5 mL) was added
intermediate 18.1 (152 mg, 0.64 mmol). Stirring was continued
overnight at rt. The mixture was diluted with crushed ice and
water. pH was adjusted to 3 by the addition of 3N HCl. The white
solid was collected and characterized as the title compound.
Compound I-29 (160 mg, 0.45 mmol) was obtained as a white powder.
Yield 70%. .sup.1H NMR (400 MHz, DMSO-d.sub.6) .delta. 4.50 (s,
2H), 6.63 (s, 1H), 7.54 (t, J=7.7 Hz, 1H), 7.64 (d, J=7.5 Hz, 1H),
7.9 (d, J=7.6 Hz, 1H), 8.13 (s, 1H), 13.3 (brs, 1H); HPLC:
95.2%.
Example 50:
(4-Benzyl-5-cyano-6-oxo-1,6-dihydro-pyrimidin-2-ylsulfanyl)-acetic
acid (Compound I-30)
##STR00321##
[0876] To a stirred solution of intermediate 4.2 (110 mg, 0.45
mmol) and DIPEA (0.086 mL, 0.5 mmol) in acetone (10 mL) was added
chloroacetic acid (43 mg, 0.45 mmol). Stirring was continued
overnight reflux. The mixture cooled to rt and it was diluted with
crushed ice and water. pH was adjusted to 3 by the addition of 3N
HCl. The aqueous phase was extracted with EtOAc (3.times.20 mL).
The combined organic phase was washed with brine and dried over
Na.sub.2SO4. The crude of reaction was purified by reverse phase
flash chromatography eluting with H.sub.2O/MeOH from 5 to 80% for
product. Compound I-30 (95 mg, 0.32 mmol) was obtained as a white
powder. Yield 69%. .sup.1H NMR (400 MHz, DMSO-d.sub.6) .delta. 3.92
(s, 2H), 4.0 (s, 2H), 7.24-7.28 (m, 1H), 7.31-7.32 (4H); .sup.13C
NMR (100 MHz, DMSO-d.sub.6) .delta. 33.4, 42.6, 95.0, 115.4, 127.4,
129, 129, 129.3, 129.3, 136.4, 160.7, 166.3, 169.4, 169.4; HPLC:
98.9%.
Example 51:
3-(6-Oxo-4-trifluoromethyl-1,6-dihydro-pyrimidin-2-ylsulfanylmethyl)-benz-
oic acid (Compound I-31)
##STR00322##
[0878] To a stirred solution of intermediate 2.1 (200 mg, 0.92
mmol) and DIPEA (0.19 mL, 1.1 mmol) in DMSO (5 mL) was added
3(2-chloromethyl) benzoic acid 13.1 (170 mg, 1 mmol). Stirring was
continued overnight at rt. The mixture was diluted with crushed ice
and water. pH was adjusted to 3 by the addition of 3N HCl. The
white precipitiate was collected and dried under vacuo. The crude
was purified by reverse phase chromatography, eluting with
H2O/MeOH. from 4 to 80%. Compound I-31 (120 mg, 0.36 mmol) was
obtained as a white powder. Yield 40%. .sup.1H NMR (400 MHz,
DMSO-d.sub.6) .delta. 2.5 (s, 2H), 4.45 (s, 2H), 6.61 (s, 1H), 7.42
(t, J=7.6 Hz, 1H), 7.48 (d, J=7.4 Hz, 1H), 7.81 (d, J=7.58 Hz, 1H),
8.02 (s, 1H), 13.12 (brs, 1H); .sup.13C NMR (100 MHz, DMSO-d.sub.6)
.delta. 33.8, 107.7, 120.6 (q, J.sub.CF=2.7 Hz), 128.6, 128.9,
130.4, 131.3, 134, 138.3, 150, 163, 165, 167.4; HPLC: 98.8%.
Example 52:
3-(6-Oxo-4-trifluoromethyl-1,6-dihydro-pyrimidin-2-ylsulfanylmethyl)-benz-
oic acid (Compound I-32)
##STR00323##
[0880] To a stirred solution of intermediate 4.2 (200 mg, 0.92
mmol) and DIPEA (0.16 mL, 0.9 mmol) in acetone (10 mL) was added 5
chloromethyl-1H-tetrazole (97 mg, 0.82 mmol). Stirring was
continued overnight at reflux. The mixture was diluted with crushed
ice and water. pH was adjusted to 3 by the addition of 3N HCl. The
mixture was extracted with EtOAc (3.times.30 mL). The combined
organic phase was washed with brine and dried over Na.sub.2SO4.
Compound I-32 (58 mg, 0.18 mmol) was obtained as a white powder
after purification by reverse phase flash chromatography, eluting
with H.sub.2O/MeOH from 5 to 80%. Yield 20%. .sup.1H NMR (400 MHz,
DMSO-d.sub.6) .delta. 3.90 (s, 2H), 4.74 (s, 2H), 7.13-7.21 (m,
5H); HPLC: 98.9%.
Example 53:
[3-(4-Benzyl-5-cyano-6-oxo-1,6-dihydro-pyrimidin-2-ylsulfanylmethyl)-phen-
yl]-acetic acid (Compound I-33)
##STR00324##
[0882] To a stirred solution of intermediate 4.2 (150 mg, 0.62
mmol) and DIPEA (0.12 mL, 0.68 mmol) in acetone (10 mL) was added
intermediate 11.2 (146 mg, 0.68 mmol). Stirring was continued
overnight reflux. The mixture cooled to rt and it was diluted with
crushed ice and water. pH was adjusted to 5 by the addition of
AcOH. The precipitate was collected and dried under vacuo. The
title compound I-33 (80 mg, 0.2 mmol) was obtained as a white
solid. Yield: 33%. .sup.1H NMR (400 MHz, DMSO-d.sub.6) .delta. 3.5
(s, 2H), 4.0 (s, 2H), 4.37 (s, 2H), 7.12-7.32 (m, 9H); .sup.13C NMR
(100 MHz, DMSO-d.sub.6) .delta. 34.2, 42.6, 54.1, 95.2, 116, 127.5,
127.8, 128.9, 129, 129.1, 129.1, 129.6, 129.6, 130.4, 135.7, 136.8,
137.5, 161, 166.7, 172.6, 173.1; HPLC: 90.2%
Example 54:
4-Benzyl-6-oxo-2-[3-(1H-tetrazol-5-yl)-benzylsulfanyl]-1,6-dihydro-pyrimi-
dine-5 carbonitrile (Compound I-34)
##STR00325##
[0884] To a stirred solution of intermediate 4.2 (150 mg, 0.62
mmol) and DIPEA (0.12 mL, 0.68 mmol) in acetone (10 mL) was added
intermediate 18.1 (162 mg, 0.68 mmol). Stirring was continued
overnight reflux. The mixture cooled to rt and it was diluted with
crushed ice and water. pH was adjusted to 5 by the addition of
AcOH. The precipitate was collected and dried under vacuo. The
crude was suspended in water, acidified to pH 3 by the addition of
3N HCl solution and extracted with EtOAc, (3.times.20 mL). The
combined organic phase was washed with brine and dried over
Na.sub.2SO.sub.4. The title compound I-34 (125 mg, 0.31 mmol) was
obtained as a light yellow solid. Yield: 50%. .sup.1H NMR (400 MHz,
DMSO-d.sub.6) .delta. 4.0 (s, 2H), 4.51 (s, 2H), 7.17-7.28 (m, 5H),
7.41-7.47 (2H), 7.90 (d, J=6.82 Hz, 1H), 8.06 (s, 2H); .sup.13C NMR
(100 MHz, DMSO-d.sub.6) .delta. 33.7, 42.5, 95.5, 115.5, 124.7,
126.2, 127.3, 128.8, 128.9, 128.9, 129.4, 129.4, 129.9, 132.1,
136.5, 139, 158, 160.6, 166.2, 172.7; HPLC: 93%.
Example 55:
3-(4-Benzyl-5-cyano-6-oxo-1,6-dihydro-pyrimidin-2-ylsulfanylmethyl)-benzo-
ic acid (Compound I-35)
##STR00326##
[0886] To a stirred solution of intermediate 4.2 (150 mg, 0.62
mmol) and DIPEA (0.14 mL, 0.82 mmol) in acetone (10 mL) was added
3(2-chloromethyl) benzoic acid 13.1 (116 mg, 0.68 mmol). Stirring
was continued overnight at r.t. The mixture cooled to rt and it was
diluted with crushed ice and water. pH was adjusted to 3 by the
addition of 3N HCl. The precipitate was collected and dried under
vacuo. The title compound I-35 (160 mg, 0.42 mmol) was obtained as
a light yellow solid. Yield: 68%. .sup.1H NMR (400 MHz,
DMSO-d.sub.6) .delta. 4.0 (s, 2H), 4.46 (s, 2H), 7.25-7.36 (m, 5H),
7.44 (d, J=7.3 Hz, 1H), 7.81 (d, J=7.6 Hz, 1H), 8.0 (s, 1H), 13.1
(brs, 1H); .sup.13C NMR (100 MHz, DMSO-d.sub.6) .delta. 33.7, 42.5,
95.5, 115.5, 127.4, 128.6, 129, 129, 129.1, 129.4, 129.4, 130.3,
131.2, 133.8, 136.5, 138.1, 160.5, 166.2, 167.4, 172.7; HPLC: 98%.
HPLC: 98%.
Example 56:
[3-(3-cyano-6-oxo-4-trifluoromethyl-1,6-dihydro-pyridin-2-ylsulfanylmethy-
l)-phenyl]-acetic acid (Compound I-36)
##STR00327##
[0888] To a stirred solution of intermediate 7.1 (164 mg, 0.63
mmol) and DIPEA (0.12 mL, 0.71 mmol) in DMSO (5 mL) was added
intermediate 11.2 (150 mg, 0.71 mmol). Stirring was continued
overnight at r.t. The mixture was diluted with crushed ice and
water. pH was adjusted to 3 by the addition of 3N HCl followed by
extraction with EtOAc, (3.times.20 mL). The combined organic phase
was washed with brine, dried over Na.sub.2SO.sub.4. The crude was
purified by reverse phase chromatography eluting with H.sub.2O/MeOH
from 8% to 40% for product. The title compound I-36 (85 mg, 0.23
mmol) was obtained as a white powder. Yield: 37%. .sup.1H NMR (400
MHz, DMSO-d.sub.6) .delta. 3.55 (s, 2H), 4.53 (s, 2H), 6.87 (s,
1H), 7.16 (m, 1H), 7.26 (m, 1H), 7.35 (m, 2H), 12.2 (brs, 1H).
.sup.13C NMR (100 MHz, DMSO-d.sub.6) .delta. 34.1, 40.8, 91.5,
105.3, 113.8, 121.5 (q, J.sub.CF=2.7 Hz), 127.8, 128.8, 129, 130.6,
135.7, 137.1, 142 (q, J.sub.CF=0.3 Hz), 164.6, 165.6, 173; HPLC:
97.8%.
Example 57:
[3-(5-Cyano-6-oxo-4-thiophen-2-yl-1,6-dihydro-pyrimidin-2-ylsulfanyl)-pro-
pionic acid (Compound I-37)
##STR00328##
[0890] To a stirred solution of intermediate 1.4 (200 mg, 0.84
mmol) and DIPEA (0.16 mL, 0.93 mmol) in acetone (10 mL) was added
3-bromo-propionic acid (57.1) (142 mg, 0.93 mmol). Stirring was
continued overnight at rt. The mixture was then diluted with
crushed ice and water and the pH was adjusted to 3 by the addition
of 3N HCl. The resulting precipitate was collected and dried under
vacuo. The title compound I-37 (180 mg, 0.58 mmol) was obtained as
a light yellow solid. Yield: 69%. .sup.1H NMR (400 MHz,
DMSO-d.sub.6) .delta. 2.75-2.80 (m, 2H), 3.34-3.39 (m, 2H),
7.32-7.36 (m, 1H), 8.0-8.24 (m, 1H), 8.24-8.27 (m, 1H), 12.18 (brs,
1H), 13.72 (brs, 1H); .sup.13C NMR (100 MHz, DMSO-d.sub.6) .delta.
26.2, 33.8, 40.7, 88.4, 1116.6, 130.1, 131.9, 135.4, 139.9, 158.9,
161.1, 165.6, 173.1; HPLC: 96.6%.
Example 58:
3-(6-Oxo-4-trifluoromethyl-1,6-dihydro-pyrimidin-2-ylsulfanyl)-propionic
acid (Compound I-38)
##STR00329##
[0892] To a stirred solution of intermediate 2.2 (150 mg, 0.69
mmol) and DIPEA (0.13 mL, 0.75 mmol) in DMSO (5 mL) was added
3-bromo-propionic acid (57.1) (115 mg, 0.75 mmol). Stirring was
continued overnight at rt. The mixture was diluted with crushed ice
and water and the pH was adjusted to 3 by the addition of 3N HCl.
The aqueous phase was extracted with EtOAc (3.times.20 mL). The
combined organic phase was washed with brine, and dried over
Na.sub.2SO.sub.4. The title compound I-38 (65 mg, 0.24 mmol) was
obtained as a white powder. Yield: 35%. .sup.1H NMR (400 MHz,
DMSO-d.sub.6) .delta. 2.69 (t, J=6.68 Hz, 2H), 3.27 (t, J=6.7 Hz,
2H), 6.61 (s, 1H), 12.9 (brs, 2H); HPLC: 98.8%.
Example 59:
2-(3,5-Difluoro-4-hydroxy-benzylsulfanyl)-6-oxo-4-thiophen-2-yl-1,6-dihyd-
ro-pyrimidine-5-carbonitrile (Compound I-39)
##STR00330##
[0893] Step 1:
2-(3,5-Difluoro-4-methoxy-benzylsulfanyl)-6-oxo-4-thiophen-2-yl-1,6-dihyd-
ro-pyrimidine-5-carbonitrile (59.2)
[0894] To a solution of intermediate 1.4 (150 mg, 0.64 mmol) in
DMSO (5 mL) was added DIPEA (0.12 mL, 0.7 mmol) and intermediate
59.1 (135 mg, 0.7 mmol). Stirring was continued at rt 16 h. The
crude was poured in water, acidified to pH 3 by a 3N solution of
HCl. The aqueous phase was extracted with EtOAc (3.times.20 mL).
The combined organic phase was washed with brine and dried over
Na.sub.2SO.sub.4. The intermediate 59.2 (168 mg, 0.42 mmol) was
obtained as a orange powder. Yield: 67%. .sup.1H NMR (400 MHz,
DMSO-d.sub.6) .delta. 3.87 (s, 2H), 4.5 (s, 3H), 7.28 (d, J=9.1 Hz,
2H), 7.36 (t, J=4.6 Hz, 1H), 8.1 (d, J=4.9 Hz, 1H), 8.28 (d, J=3.8
Hz, 1H).
Step 2:
2-(3,5-Difluoro-4-hydroxy-benzylsulfanyl)-6-oxo-4-thiophen-2-yl-1,-
6-dihydro-pyrimidine-5-carbonitrile (I-39)
[0895] To a stirred suspension of intermediate 59.2 (160 mg, 0.41
mmol) in DCM (10 mL) was added at 0.degree. C. a 1 M solution of
BBr.sub.3 in DCM (0.5 mL, 0.45 mmol). Stirring was continued at rt
16 h at rt. The reaction was quenched with MeOH and the solvents
were removed under vacuo. The crude was purified by flash
chromatography eluting with DCM/MeOH from 0 to 6% for product. The
title compound I-39 (68 mg, 0.18 mmol) was obtained as a white
powder after trituration with Et.sub.2O. Yield: 44%. .sup.1H NMR
(400 MHz, DMSO-d.sub.6) .delta. 4.47 (s, 2H), 7.17 (d, J=8.3 Hz,
2H), 7.36 (t, J=4.6 Hz, 1H), 8.28 (d, J=3.9 Hz, 1H), 10.25 (s, 1H),
13.9 (brs, 1H); .sup.13C NMR (100 MHz, DMSO-d.sub.6) .delta. 33.3,
88.7, 112.7 (d, J.sub.CF=28.6 Hz), 112.9 (d, J.sub.CF=28.6 Hz),
116.5, 127.8 (t, J.sub.CF=32.8 Hz), 130, 132.1, 133.4 (t,
J.sub.CF=63.7 Hz), 135.4, 139.7, 151.0 (d, J.sub.CF=28.9 Hz), 153.4
(d, J.sub.CF=28.6 Hz), 158.9, 161.2, 165.2; HPLC: 95.7%.
Example 60:
2-(3,5-Difluoro-4-hydroxy-benzylsulfanyl)-6-trifluoromethyl-3H-pyrimidin--
4-one (Compound I-40)
##STR00331##
[0896] Step 1:
2-(3,5-Difluoro-4-methoxy-benzylsulfanyl)-6-trifluoromethyl-3H-pyrimidin--
4-one (60.1)
[0897] Following the procedure of Example 59 (Step 1) and starting
from intermediate 2.2 (200 mg, 0.91 mmol), 59.1 (211.8 mg, 1.1
mmol) and DIPEA (0.19 mL, 1.1 mmol) in DMSO (5 mL) the title
intermediate 60.1 (200 mg, 0.56 mmol) was obtained as white powder.
Yield 62%. .sup.1H NMR (400 MHz, DMSO-d.sub.6) .delta. 3.88 (s,
2H), 4.33 (s, 3H), 6.63 (s, 1H), 7.21 (d, J=9.2 Hz, 2H).
Step 2:
2-(3,5-Difluoro-4-hydroxy-benzylsulfanyl)-6-trifluoromethyl-3H-pyr-
imidin-4-one (I-40)
[0898] To a stirred suspension of intermediate 60.1 (190 mg, 0.54
mmol) in DCM (10 mL) was added at 0.degree. C. a 1 M solution of
BBr.sub.3 in DCM (0.6 mL, 0.59 mmol). Stirring was continued at rt
16 h at rt. The reaction was quenched with MeOH. The solvents were
removed under vacuo. The crude was purified by flash chromatography
eluting with DCM/MeOH from 0 to 6% for product. The title compound
I-40 (61 mg, 0.18 mmol) was obtained as a white powder after
trituration with Et.sub.2O. Yield: 33%. .sup.1H NMR (400 MHz,
DMSO-d.sub.6) 4.29 (s, 2H), 6.63 (s, 1H), 7.11 (d, J=7.82 Hz, 2H),
10.19 (s, 1H), 13.15 (brs, 1H); .sup.13C NMR (100 MHz,
DMSO-d.sub.6) .delta. 33.3, 113,1 (d, J.sub.CF=28.2 Hz), 113.2 (d,
J.sub.CF=28.4 Hz), 119.6, 122.3, 128.2, 133.3 (t, J.sub.CF=64.1
Hz), 150.9 (d, J.sub.CF=28.3 Hz), 153.3 (d, J.sub.CF=28.4 Hz);
HPLC: 98.3%.
Example 61:
4-Benzyl-2-(3,5-difluoro-4-hydroxy-benzylsulfanyl)-6-oxo-1,6-dihydro-pyri-
midine-5-carbonitrile (Compound I-41)
##STR00332##
[0899] Step 1:
4-Benzyl-2-(3,5-difluoro-4-methoxy-benzylsulfanyl)-6-oxo-1,6-dihydro-pyri-
midine-5-carbonitrile (61.1)
[0900] Following the procedure of Example 59 (Step 1) and starting
from intermediate 4.2 (200 mg, 0.82 mmol), 59.1 (189.9 mg, 0.98
mmol) and DIPEA (0.17 mL, 0.98 mmol) in DMSO (5 mL) the title
intermediate 60.1 (200 mg, 0.5 mmol) was obtained as white powder.
Yield 61%. .sup.1H NMR (400 MHz, DMSO-d.sub.6) .delta. 3.87 (s,
2H), 4.02 (s, 2H), 4.35 (s, 2H), 7.0 (d, J=8.9 Hz, 2H), 7.3 (m,
5H).
Step 2:
4-Benzyl-2-(3,5-difluoro-4-hydroxy-benzylsulfanyl)-6-oxo-1,6-dihyd-
ro-pyrimidine-5-carbonitrile (Compound I-41)
[0901] To a stirred suspension of intermediate 61.1 (190 mg, 0.54
mmol) in DCM (10 mL) was added at 0.degree. C. a 1 M solution of
BBr.sub.3 in DCM (0.5 mL, 0.52 mmol). Stirring was continued at rt
16 h at rt. The reaction was quenched with MeOH. The solvents were
removed under vacuo. The crude was purified by flash chromatography
eluting with DCM/MeOH from 0 to 6% for product. The title compound
I-41 (40 mg, 0.1 mmol) was obtained as a white powder after
trituration with Et.sub.2O. Yield: 22%. .sup.1H NMR (400 MHz,
DMSO-d.sub.6) 4.02 (s, 2H), 4.3 (s, 2H), 6.93 (d, J=7.6 Hz, 2H),
7.27 (m, 4H), 7.9 (brs, 1H), 10.2 (s, 1H), 13.92 (bras, 1H); HPLC
96.7%.
Example 62:
6-Oxo-2-{[4-(1H-tetrazol-5-yl)-cyclohexylmethyl]-amino}-4-thiophen-2-yl-1-
,6-dihydro-pyrimidine-5-carbonitrile (Compound I-42)
##STR00333##
[0902] Step 1: (4-Carbamoyl-cyclohexylmethyl)-carbamic acid
tert-butyl ester (62.2)
[0903] To a solution of the starting intermediate 62.1 (1.3 g, 5.1
mmol) in CH.sub.3CN (20 mL) was added pyridine (0.45 mL, 5.55
mmol), Boc.sub.2O (1.67 g, 7.65 mmol) and ammonium bicarbonate (605
mg, 7.65 mmol). Stirring was continued at rt 16 h. The crude was
poured in water. The aqueous phase was extracted with EtOAc
(3.times.20 mL). The combined organic phase was washed with brine,
and dried over Na.sub.2SO.sub.4. The title compound 62.2 (1 g, 3.9
mmol) was obtained as a white powder. Yield: 76%. .sup.1H NMR (400
MHz, CDCl.sub.3) .delta. 0.8 (q, J=2.6 Hz, 2H), 1.45 (s, 9H), 1.63
(s, 2H), 1.86 (d, J=13.3 Hz, 2H), 1.97 (d, J=13.4 Hz, 2H), 2.11 (t,
J=3.4 Hz, 1H), 2.99 (m, 2H), 4.5 (s, 1H), 5.38 (m, 2H).
Step 2: (4-Cyano-cyclohexylmethyl)-carbamic acid tert-butyl ester
(62.3)
[0904] A solution of the starting intermediate 62.2 (200 mg, 0.78
mmol) in DCM (10 ml) was added Et3N (0.3 mL, 1.95 mmol). The
mixture was cooled at 0.degree. C. and TFAA (0.14 mL, 0.98 mmol)
was added dropwise. Stirring was continued at rt 16h. The reaction
was poured in water, extracted with DCM (3.times.20 mL). The
combined organic phase was washed with brine and were dried over
Na.sub.2SO.sub.4 to give the title compound 62.3 (150 mg, 0.62
mmol) as yellow oil. Yield 84%. .sup.1H NMR (400 MHz, CDCl.sub.3)
.delta. 0.98-1 (m, 2H), 1.46 (s, 9H), 1.52 (m, 2H), 1.57-1.61 (m,
1H), 1.75-1.77 (m, 1H), 1.83-1.87 (m, 2H), 2.12-2.16 (m, 2H),
2.35-2.42 (m, 1H), 2.98 (t, J=6.4 Hz, 2H), 3.58 (d, J=6.8 Hz, 1H),
4.61 (brs, 1H).
Step 3: [4-(1H-Tetrazol-5-yl)-cyclohexylmethyl]-carbamic acid
tert-butyl ester (62.4)
[0905] To a solution of the starting intermediate 62.3 (200 mg,
0.84 mmol) in DMF (3 mL) was sodium azide (164 mg, 2.52 mmol) and
NH4Cl (135 mg, 2.52 mmol). Stirring was continued at 140.degree. C.
for 25h. The crude was poured in water, acidified to pH3 by the
addition of 3M HCl solution. The aqueous phase was extracted with
EtOAc (3.times.20 mL). The combined organic phase was washed with
brine and dried over Na.sub.2SO.sub.4. The title compound 62.4 (160
mg, 0.56 mmol) was obtained as a white powder. Yield: 67%. .sup.1H
NMR (200 MHz, DMSO-d.sub.6) .delta. 1.01 (t, J=6.8 Hz, 2H),
1.36-1.52 (m, 11H), 1.76 (d, J=11.7 Hz, 2H), 2.01 (d, J=11.0 Hz,
2H), 2.72-2.96 (m, 3H), 6.59-6.87 (m, 1H), 14.1 (brs, 1H).
Step 4: C-[4-(1H-Tetrazol-5-yl)-cyclohexyl]-methylamine (62.5)
[0906] To a solution of intermediate 62.4 (450 mg, 1.60 mmol) in
dioxane (5 mL) was added a 4M solution of HCl in dioxane (13.2 mL).
Stirring was continued at r.t 16h. The solvent was removed under
vacuo to give the title intermediate 62.5 (378 mg, 1.73 mmol) as a
white powder chlorohydrate salt. Yield 95%. .sup.1H NMR (200 MHz,
DMSO-d.sub.6) .delta. 1.0-1.14 (m, 2H), 1.46-1.52 (m, 3H), 1.84 (d,
J=13.1 Hz, 2H), 2.01 (d, J=13.4 Hz, 2H), 2.64-2.70 (m, 2H), 2.93
(m, 1H), 8.05 (brs, 3H).
Step 5:
6-Oxo-2-{[4-(1H-tetrazol-5-yl)-cyclohexylmethyl]-amino}-4-thiophen-
-2-yl-1,6-dihydro-pyrimidine-5-carbonitrile (Compound I-42)
[0907] To a stirred solution of intermediate 62.6 (250 mg, 0.88
mmol) in DMSO (5 mL) was added DIPEA (0.3 mL, 1.76 mmol) and
intermediate 62.5 (211 mg, 0.97 mmol). Stirring was continued at
80.degree. C. 4 h. The crude was poured in water, acidified to pH 3
and extracted with EtOAc (3.times.20 mL). The crude product from
the reaction was purified by flash chromatography eluting with
DCM/MeOH (6% for product). The title compound I-42 (24 mg, 0.06
mmol) was obtained as yellowish solid. Yield 7%. MS-ESI (-) m/z:
381.4 (M-H). HPLC: 88%
Example 63: Preparation of Intermediate 1b.3
##STR00334##
[0908] Step 1: Preparation of Intermediate 1b.2
[0909] To a stirred solution of compound 1b.1 (500 mg, 2.13 mmol),
in a mixture of H.sub.2O/EtOH (2 mL+4 mL) was added NaOH (85 mg,
2.13 mmol) and Mel (0.12 mL, 2.13 mmol). Stirring was continued at
60.degree. C. for 30 minutes. The title compound 1b.2 was collected
as yellow powder upon filtration from the reaction medium (491 mg,
4.9 mmol). Yield 92%.
Step 2: Preparation of Intermediate 1b.3
[0910] To a stirred solution of compound 1b.2 (200 mg, 0.8 mmol),
in CHCl.sub.3 (6 mL) was added mCPBA (207 mg, 1.2 mmol). Stirring
was continued at rt for 16h. The yellow solid was collected and
washed with DCM and Et.sub.2O. The title compound 1b.3 was obtained
(190 mg, 0.67 mmol) as a pale yellow solid. Yield 84%.
Example 64: Preparation of Intermediate 2h.2
##STR00335##
[0911] Step 1: Preparation of Intermediate 2b.2
[0912] To a solution of the starting intermediate 2b.1 (200 mg,
0.83 mmol) in MeOH (3 mL) was added a 30% ammonia solution in water
(2 mL). Stirring was continued at rt for 24h. The solvent was then
removed under vacuo. The title intermediate 2b.2 (143 mg, 0.74
mmol) was obtained as yellowish powder without further
purifications. Yield 90%.
Example 65: Preparation of Intermediate 3b.6
##STR00336##
[0913] Step 1: Preparation of Intermediate 3b.2
[0914] To a stirred and boiling solution of 3b.1 (1.00 g, 7.62
mmol) in MeCN (30 mL), a solution of NBS (1.42 g) and BPO 70% (13
mg, 0.04 mmol) in MeCN (10 mL) was added dropwise. After 30 min the
mixture was slowly cooled to r.t. and poured in aq. NaHCO.sub.3 ss
(15 mL). The mixture was extracted with AcOEt (3.times.30 mL),
washed with brine (50 mL), dried over Na.sub.2SO4, and concentrated
under reduced pressure. The title intermediate 3b.2 was used
directly (1.60 g) for the next step.
Step 2: Preparation of Intermediate 3b.3
[0915] A solution of intermediate 3b.2 (1.60 g, crude of previous
step) in MeOH (15 mL) was treated with aq. NH.sub.3 28% (15 mL),
and the resulting mixture was stirred for 16 h. Volatiles were
removed under reduced pressure and the crude was poured in H.sub.2O
(30 mL) and washed with AcOEt (3.times.30 mL). The aqueous phase
was concentrated under reduced pressure. The title intermediate
3b.3 (1.17 g) was used as crude for the next step.
Step 3: Preparation of Intermediate 3b.4
[0916] A stirred suspension of intermediate 3b.3 (1.17 g, crude of
previous step) in CH.sub.2Cl.sub.2 (30 mL), Boc.sub.2O (1.99 g,
9.14 mmol) and DIPEA (3.32 mL, 19.06 mmol) were added, and the
mixture was reacted at r.t. for 2 h obtaining an opalescent
solution. H.sub.2O (20 mL) was added, the two phases were separated
and the organic one was washed with aq. citric acid 0.5 M
(2.times.20 mL), H.sub.2O (20 mL), brine (20 mL), dried over
Na.sub.2SO.sub.4 and concentrated under reduced pressure. The crude
was purified by chromatographic purification (Petroleum ether/AcOEt
from 9:1 to 7:3) to give the title intermediate 3b.4 as dense oil
in 28% yield starting from 3.1. MS-ESI (+) m/z: 247.2 (M+H).
Step 4: Preparation of Intermediate 3b.5
[0917] A mixture of intermediate 3b.4 (525 mg, 2.13 mmol), sodium
azide (415 mg, 6.39 mmol) and triethylammonium chloride (880 mg,
6.39 mmol) in toluene (40 mL) was stirred and refluxed for 18 h.
Once cooled at r.t., aq. NaHCO.sub.3 ss (15 mL) was added, and the
mixture was vigorously stirred for 10 min. The two phases were
separated and the organic one was extracted with H.sub.2O
(3.times.30). All the aqueous phases were collected together and
acidified up to pH=3 by adding citric acid 0.5 M. The resulting
acid aqueous phase was extracted with CH.sub.2Cl.sub.2 (3.times.50
mL), washed with), brine (50 mL), dried over Na.sub.2SO.sub.4 and
concentrated under reduced pressure. The title intermediate 3b.5
(214 mg, 0.74 mmol) was obtained as white solid. Yield: 35%. MS-ESI
(-) m/z: 288.2 (M-H).
Step 5: Preparation of Intermediate 3b.6
[0918] Intermediate 3b.5 (205 mg, 0.71 mmol) was dissolved in MeOH
(10 mL) and treated with HCl 37% (0.29 mL, 3.35 mmol) at 50.degree.
C. for 2 h. Volatiles were removed under reduced pressure, to
afford intermediate 3b.6 in nearly quantitative yield as
hydrochloride salt. MS-ESI (+) m/z: 190.3 (M+H); MS-ESI (-) m/z:
188.2 (M-H).
Example 66: Preparation of Intermediate 4b.4
##STR00337##
[0919] Step 1: Preparation of Intermediate 4b.2
[0920] To a solution of intermediate 4b.1 (2 g, 11.86 mmol) in DCM
(20 mL) was added TEA (1.99 mL, 14.2 mmol) and BOC.sub.2O (2.717 g,
12.45 mmol). Stirring was continued at rt 5h. The crude was poured
in water and was extracted with DCM. The organic phase were washed
with brine and dried over Na.sub.2SO.sub.4. The title intermediate
4b.2 (2 g, 8.61 mmol) was obtained as white solid. Yield 72%.
Step 2: Preparation of Intermediate 4b.3
[0921] To a solution of the starting intermediate 4b.2 (2 g, 7.26
mmol) in DMF (6 mL) was added NaN.sub.3 (839 mg, 12.91 mmol) and
ammonium chloride (689 mg, 12.91 mmol). Stirring was continued at
140.degree. C. for 6 h. The crude was poured in water and brine
followed by extraction with EtOAc at pH=3. The organic phase were
dried over Na.sub.2SO.sub.4 and evaporated under vacuo. The title
intermediate 4b.3 (2.2 g, 8.0 mmol) was obtained as white solid.
Yield 93%.
Step 3: Preparation of Intermediate 4b.4
[0922] The starting intermediate 4b.3 (1.5 g, 5.48 mmol) was
stirred overnight in a 4 M dioxane solution of HCl (10 mL). The
solvent was removed under vacuo. The title intermediate 4b.4 (1.14
g, 5.38 mmol) was obtained as a white solid. Yield 98%.
Example 67: Preparation of Intermediate 5b.2
##STR00338##
[0924] To a solution of intermediate 5b.1 (1 g, 6.1 mmol) in EtOH
(10 mL) was added NH.sub.2CN (0.7 mL, 9.1 mmol) and HNO.sub.3 (0.25
mL, 6.1 mmol). Stirring was continued at reflux for 16 h. The
mixture was cooled to 0.degree. C. and was added of Et20. The white
precipitate was collected. The title intermediate 5b.2 (1 g, 3.7
mmol) was thus obtained as white solid as HNO.sub.3 salt. Yield
60%.
Example 68: Preparation of Intermediate 6b.2
##STR00339##
[0926] To a solution of intermediate 6b.1 (1 g, 6.1 mmol) in EtOH
(10 mL) was added NH.sub.2CN (0.7 mL, 9.1 mmol) and HNO.sub.3 (0.25
mL, 6.1 mmol). Stirring was continued at reflux for 16 h. The
mixture was cooled to 0.degree. C. and was added of Et20. The white
precipitate was collected. The title intermediate 6b.2 (1.2 g, 4.4
mmol) was obtained as yellowish solid as HNO.sub.3 salt. Yield
72%.
Example 69: Preparation of Compound I-43
##STR00340##
[0928] To a stirred suspension of intermediate 1b.3 (200 mg, 0.85
mmol) in DMSO (5 mL) was added DIPEA (0.29 mL, 1.7 mmol) and
intermediate 2b.2 (164 mg, 0.94 mmol). Stirring was continued at
80.degree. C. for 4 h. The crude was poured in water, acidified to
pH 3 and extracted with EtOAc (3.times.20 mL). The crude of
reaction was purified by flash chromatography eluting with DCM/MeOH
(3% for product). The title compound I-43 (50 mg, 0.13 mmol) was
obtained as yellowish solid. Yield 15%. .sup.1H NMR (400 MHz, DMSO
d.sub.6) .delta. 4.69 (2H), 7.25 (m, 1H), 7.58 (m, 2H), 7.91 (m,
2H), 8.0 (s, 1H), 8.16 (m, 1H), 11.94 (brs, 1H); .sup.13C NMR (100
MHz, DMSO d.sub.6) .delta. 44.2, 81.8, 117.8, 124.6, 126.1, 126.3,
129.3, 129.9, 130.7, 130.9, 130.9, 133.9, 140.5, 141.1, 154.4,
161.6, 162.1. HPLC 96.3%.
Example 70: Preparation of Compound I-44
##STR00341##
[0929] Step 1: Synthesis of Intermediate 8b.2
[0930] A solution of intermediate 8b.1 (195 mg, 0.72 mmol) and
intermediate 5b.2 (150 mg, 0.72 mmol) in DMF (2 mL), was added of
piperidine (0.14 mL, 1.45 mmol). The mixture was sealed in a Q-tube
apparatus and heated at 120.degree. C. 16h. The mixture was cooled
to rt, poured in water, extracted with EtOAc, and purified by flash
chromatography eluting with DCM/MeOH (5% for product). The title
intermediate 8b.2 (200 mg, 0.39 mmol) has been obtained as brownish
powder. Yield 54%
Step 2: Synthesis of Compound I-54
[0931] To a solution of intermediate 8b.2 (150 mg, 0.41 mmol) in
EtOH (10 mL) was added a 1 M solution of NaOH (1.2 mL). Stirring
was continued at reflux gently for 16 h. The precipitate was
collected by filtration and it was dissolved in water. The pH was
adjusted to 3 by the addition of 3N HCl solution. The precipitate
was collected and dried under vacuo to give the title compound I-44
(100 mg, 0.295 mmol) as a brown solid. Yield 72%. .sup.1H NMR (400
MHz, DMSO d.sub.6) .delta. 7.31 (t, J=4.22 Hz, 1H), 7.50 (t, J=7.8
Hz, 1H), 7.72 (d, J=7.48 Hz, 1H), 7.88 (d, J=7.4 Hz, 1H), 7.99 (d,
J=4.7 Hz, 1H), 8.25 (m, 2H), 10.1 (s, 1H), 11.9 (brs, 1H); .sup.13C
NMR (100 MHz, DMSO d.sub.6) .delta. 83.5, 117.4, 122.4, 125.3,
125.7, 129.5, 129.6, 131.2, 131.8, 134.3, 137.9, 140.8, 152.5,
161.4, 161.9, 167.3. HPLC: 98.4%
Example 71: Preparation of Compound I-45
##STR00342##
[0932] Step 1: Synthesis of Intermediate 9b.1
[0933] A solution of intermediate 8b.1 (990 mg, 0.72 mmol) and
intermediate 6b.2 (1.18 g, 4.34 mmol) in DMF (10 mL), was added of
piperidine (0.86 mL, 8.68 mmol). The mixture was heated at
150.degree. C. for 16h. The mixture was cooled to rt, poured in
water, the pH was adjusted to 3 by the addition of HCl (3N). The
solid was collected and washed with acetone. The title intermediate
9b.1 (250 mg, 0.68 mmol) has been obtained as grey powder. Yield
14%
Step 2: Synthesis of Compound I-45
[0934] To a solution of intermediate 9b.1 (150 mg, 0.41 mmol) in
EtOH (10 mL) was added a 1 M solution of NaOH (1.2 mL). Stirring
was continued at reflux gently 16 h. The solvent was removed under
vacuo. The solid was suspended in EtOH (5 mL) sonicated and
filtered. The sodium salt was dissolved in water and ice, pH was
adjusted to 3 by the addition of 3N HCl solution. The gummy
precipitate was collected and dried under vacuo to give the title
compound I-45 (90 mg, 0.21 mmol) brown solid. Yield 65%. .sup.1H
NMR (400 MHz, DMSO d.sub.6) .delta. 7.32 (d, J=4.1 Hz, 1H), 7.79
(d, J=8.6 Hz, 2H), 7.96 (d, J=8.6 Hz, 2H), 8.0 (d, J=4.9 Hz, 1H),
8.24 (d, J=3.8 Hz, 1H), 10.23 (s, 1H), 11.9 (brs, 1H); .sup.13C NMR
(100 MHz, DMSO d.sub.6) .delta. 83.9, 117.2, 120.5, 120.5, 126.2,
129.7, 130.7, 130.7, 131.3, 134.5, 140.7, 141.9, 152.5, 161.3,
167.2; HPLC: 95.56%.
Example 72: Preparation of Compound I-46
##STR00343##
[0936] To a solution of intermediate 1b.3 (100 mg, 0.36 mmol) in
DMSO (3 mL) was added 1,4-trans amino cyclohexane (51 mg, 0.36
mmol). Stirring was continued at 80.degree. C. for 16 h. The
mixture was poured in water, the pH was adjusted to 3 by the
addition of HCl (3N solution). The gummy precipitate was collected
and dried under vacuo. The crude was purified by flash
chromatography eluting with DCM/MeOH 15% and acetone due to the low
compound solubility. The title compound I-46 (40 mg, 0.12 mmol) was
obtained as yellowish powder. Yield 32%. .sup.1H NMR (200 MHz, DMSO
d.sub.6) .delta. 1.34 (m, 4H), 1.92 (m, 4H), 2.20 (brs, 1H), 3.73
(brs, 1H), 7.27 (t, J=4.2 Hz, 1H), 7.61 (s, 1H), 7.91 (d, J=4.9 Hz,
1H), 8.15 (m, 1H), 12.0 (brs, 1H). .sup.13C NMR (100 MHz, DMSO
d.sub.6) .delta. 27.8, 27.8, 31.1, 31.1, 41.7, 41.7, 50.2, 81.05,
118.0, 129.4, 130.7, 133.9, 141.3, 153.9, 161.6, 176.8; HPLC:
96.3%.
Example 73: Preparation of Compound I-47
##STR00344##
[0938] To a solution of intermediate 1b.3 (150 mg, 0.53 mmol) in
DMSO (5 mL) was added 1,4-cis amino cyclohexane (76 mg, 0.53 mmol).
Stirring was continued at 80.degree. C. for 16 h. Ice was added to
the mixture under stirring. The white solid was collected and then
purified by flash chromatography, eluting with DCM/MeOH. The title
compound I-47 (70 mg, 0.2 mmol) was obtained as white solid. Yield
38%. .sup.1H NMR (400 MHz, DMSO d.sub.6) .delta. 1.71 (m, 8H), 2.4
(brs, 1H), 4.05 (s, 1H), 7.28 (t, J=4.2 Hz, 1H), 7.3 (m, 1H), 7.93
(d, J=4.6 Hz, 1H), 8.18 (d, J=3.23 Hz, 1H), 10.9 (brs, 1H), 12.1
(brs, 1H); .sup.13C NMR (100 MHz, DMSO d.sub.6) .delta. 24.6, 28.8,
47.6, 48.9, 55.3, 55.3, 81.2, 117.8, 129.4, 130.8, 133.9, 141.2,
153.6, 161.6, 161.7, 176.5; HPLC: 98.51%.
Example 74: Preparation of Compound I-48
##STR00345##
[0939] Step 1: Synthesis of Intermediate 12b.2
[0940] To a solution of the starting intermediate 1b.3 (400 mg,
1.42 mmol) in DMSO (5 mL) was added DIPEA (0.36 mL, 2.13 mmol) and
intermediate 12b.1 (344 mg, 1.7 mmol). Stirring was continued at
80.degree. C. for 16 h. The crude was poured in water. The solution
was adjusted to pH3 by the addition of HCl (3N solution). The
aqueous phase was extracted with EtOAc. The mixture was purified by
flash chromatography, eluting with DCM/MeOH. The title intermediate
12b.2 (200 mg, 0.54 mmol) was obtained as yellowish powder. Yield
38%.
Step 2: Synthesis of Compound I-48
[0941] To a solution of intermediate 12b.2 (200 mg, 0.52 mmol) in
MeOH (15 mL) was added a 1 M solution of NaOH (3 mL). Stirring was
continued at reflux gently for 16 h. The solvent was removed under
vacuo. The sodium salt was dissolved in water and ice, pH was
adjusted to 3 by the addition of 3N HCl solution. The gelly
precipitate was collected and dried under vacuo to give the title
compound I-48 (150 mg, 0.42 mmol) as yellowish solid after
trituration with Et.sub.2O. Yield 82%. MS-ESI (+) m/z: 353.3
(M+H).
Example 75: Preparation of Compound I-49
##STR00346##
[0943] To a stirred solution of Intermediate 1b.3 (150 mg, 0.53
mmol) in DMF (10 mL), Intermediate 3b.6 (120 mg, 0.53 mmol) and
DIPEA (0.46 mL, 2.65 mmol) were added, and the mixture was reacted
at 105.degree. C. for 6 h. Once cooled at r.t. it was poured in
H.sub.2O (25 mL) and washed with Et.sub.2O (2.times.20 mL). HCl 3.0
M was added to the aqueous solution up to pH=1 and the mixture was
extracted with CH.sub.2Cl.sub.2/MeOH 9:1 (vol/vol, 3.times.30 mL).
The collected organic phases were concentrated under reduced
pressure, and the crude was purified by RP-flash chromatography
(H.sub.2O/MeCN from 8:2 to 1:9). The collected impure compound (15
mg) was tritured with cold acetone, to afford 10 mg of pure
Compound I-49 (Yield: 5%). MS-ESI (-) m/z: 389.4 (M-H). .sup.1H NMR
(400 MHz, DMSO d.sub.6) .delta. 4.25 (s, 2H), 4.55 (d, J=4.74 Hz,
2H), 7.17 (s, 1H), 7.26 (m, 4H), 7.91 (d, J=4.8 Hz, 1H), 7.94 (brs,
1H), 8.16 (d, J=3.3 Hz, 1H); .sup.13C NMR (100 MHz, DMSO d.sub.6)
.delta. 29.4, 44.3, 81.5, 117.9, 126.5, 127.9, 128.2, 129.1, 129.3,
130.8, 133.9, 136.8, 139.4, 141.1, 154.6, 155.8, 161.6, 162.4;
HPLC: 99.5%.
Example 76: Preparation of Compound I-50
##STR00347##
[0944] Step 1: Synthesis of Intermediate 14b.2
[0945] To a solution of intermediate 1b.3 (200 mg, 0.71 mmol) in
DMSO (5 mL) was added DIPEA (0.18 mL, 1.07 mmol) and intermediate
14b.1 (164 mg, 0.85 mmol). Stirring was continued at 80.degree. C.
for 16 h. The mixture was poured in water and extracted with EtOAc
(3.times.20 mL). The mixture was purified by flash chromatography
eluting with DCM/MeOH 1.5% for product. The title intermediate
14b.2 (173 mg, 0.43 mmol) was obtained as yellow solid. Yield 62%.
.sup.1H NMR (400 MHz, DMSO d.sub.6) .delta. 1.13 (t, J=7.1 Hz, 3H),
3.63 (s, 2H), 4.02 (q, J=7.0 Hz, 2H), 4.56 (d, J=5.8 Hz, 2H), 7.16
(m, 1H), 7.27 (m, 4H), 7.93 (m, 1H), 7.94 (d, J=4.0 Hz, 1H), 8.17
(d, J=3.8 Hz, 1H), 11.91 (brs, 1H).
Step 2: Synthesis of Compound I-50
[0946] To a solution of intermediate 14b.2 (165 mg, 0.42 mmol) in
EtOH (6 mL) was added a 1 M solution of NaOH (1.3 mL). Stirring was
continued at reflux gently 16 h. The mixture was cooled to room
temperature and the precipitate was collected. The sodium salt was
dissolved in water and ice, pH was adjusted to 1 by the addition of
3N HCl solution. The precipitate was collected and dried under
vacuo to give the title compound I-50 (75 mg, 0.21 mmol) as white
solid. Yield 49%. .sup.1H NMR (400 MHz, DMSO d.sub.6) .delta. 3.55
(s, 2H), 4.57 (d, J=5.8 Hz, 2H), 7.16 (m, 1H), 7.27 (m, 4H), 7.84
(m, 1H), 7.92 (d, J=4.5 Hz, 1H), 8.17 (d, J=3.7 Hz, 1H), 11.73
(brs, 1H), 12.49 (brs, 1H); .sup.13C NMR (100 MHz, DMSO d.sub.6)
.delta. 41.0, 44.3, 81.6, 117.8, 126.3, 128.7, 129.1, 129.1, 129.3,
130.9, 134.0, 135.5, 138.8, 141.1, 154.3, 161.7, 161.9, 173.0;
HPLC: 95.2%.
Example 77: Preparation of Compound I-51
##STR00348##
[0948] To a solution of intermediate 4b.2 (211.6 mg, 1 mmol) in
DMSO (5 mL) was added DIPEA and stirring was continued at rt 10
minutes. Then a solution of compound 1b.3 in DMSO was added.
Stirring was continued at 90.degree. C. for 16h. The crude was
collected, dried and purified by flash chromatography eluting with
DCM/MeOH-6% for product. The title compound I-51 was obtained (25
mg, 0.06 mmol) as yellow solid after trituration with EtOAc. Yield
7%. .sup.1H NMR (400 MHz, DMSO d.sub.6) .delta. 4.68 (d, J=5.7 Hz,
2H), 7.27 (t, J=4.1 Hz, 1H), 7.60 (d, J=7.7 Hz, 2H), 7.9 (d, J=4.9
Hz, 1H), 8.0 (d, J=7.7 Hz, 2H), 8.18 (d, J=3.6 Hz, 1H), 12.1 (brs,
1H); .sup.13C NMR (100 MHz, DMSO d.sub.6) .delta. 44.2, 53.9, 81.7,
117.8, 123.6, 127.4, 127.4, 128.7, 129.3, 130.9, 134.0, 141.0,
142.2, 154.5, 155.7, 161.6, 162.1; HPLC: 96.6%.
[0949] Biological Activity
Biological Example 1: Determination of ACMSD1 Inhibition
[0950] The activity of compounds 1-36 as inhibitors of ACMSD1 was
determined by measuring the conversion of 30H-Anthranilic Acid into
product (i.e., ACMS) in a spectrophotometrical in vitro assay.
[0951] The pre-assay mixture consisting of 3-hydroxyanthranilic
acid (3OH-HA), 3-hydroxyanthranilic acid, 3,4-diOxygenase (HAO),
and a dialyzed crude extract of E. coli BL21 (DE3) cells expressing
the recombinant enzyme, was incubated at 25.degree. C. with
monitoring of the increase in absorbance at 360 nm due to the
formation of ACMS from 30H-HA. After the reaction was completed
within .about.2 mins, an aliquot of ACMSD1 solution (prepared and
purified from Pichia Pastoris overexpressing the recombinant
enzyme) was added, and the decrease in absorbance at 360 nm was
followed at 15 second intervals. The effect of ACMS concentration
on the enzyme activity was investigated by varying 30H-HA
concentration from 2 to 20 .mu.M. Kinetic parameters were
calculated from the initial velocity data by using the
Lineweaver-Burk plot.
[0952] The rate of the decrease in absorbance caused by ACMSD1 was
calculated by subtracting that of the control reaction mixture
without ACMSD from that described above. One unit of ACMSD activity
was indicated as the amount of enzyme that converts 1 mmol of ACMS
per minute at 25.degree. C. The absence or a reduction of ACMSD1
activity (e.g., by using ACMSD inhibitors) results in a slow
ACMS-spontaneous degradation (i.e., cyclization to form quinolic
acid).
[0953] The enzymatic activity was determined at a HAA concentration
of 10 .mu.M in the presence of the compounds in Table 1 below. The
compounds were tested at the concentration of about 5 .mu.M and 10
.mu.M and the IC.sub.50 was calculated for compounds showing
inhibitory activity higher than 50%. The results are shown in Table
1. In Table 1, A is <0.1 .mu.M; B is 0.1 to 1 .mu.M; C is 1 to
10 .mu.M; and D is >10 .mu.M.
TABLE-US-00006 TABLE 1 Activity Cpd No. Structure hACMSD IC.sub.50
I-1 ##STR00349## C I-2 ##STR00350## B I-3 ##STR00351## A I-4
##STR00352## C I-5 ##STR00353## B I-6 ##STR00354## C I-7
##STR00355## B I-8 ##STR00356## B I-9 ##STR00357## B I-10
##STR00358## C I-11 ##STR00359## C I-12 ##STR00360## C I-13
##STR00361## C I-14 ##STR00362## C I-15 ##STR00363## C I-16
##STR00364## D I-17 ##STR00365## D I-18 ##STR00366## B I-19
##STR00367## A I-20 ##STR00368## A I-21 ##STR00369## A I-22
##STR00370## A I-23 ##STR00371## C I-24 ##STR00372## C I-25
##STR00373## A I-26 ##STR00374## A I-27 ##STR00375## B I-28
##STR00376## B I-29 ##STR00377## A I-30 ##STR00378## B I-31
##STR00379## C I-32 ##STR00380## A I-33 ##STR00381## A I-34
##STR00382## A I-35 ##STR00383## B I-36 ##STR00384## B I-37
##STR00385## C I-38 ##STR00386## n/a I-39 ##STR00387## C I-40
##STR00388## n/a I-41 ##STR00389## C I-42 ##STR00390## n/a I-43
##STR00391## A I-44 ##STR00392## n/a I-45 ##STR00393## n/a I-46
##STR00394## D I-47 ##STR00395## n/a I-48 ##STR00396## C I-49
##STR00397## A I-50 ##STR00398## C I-51 ##STR00399## B
Biological Example 2: Determination of ACMSD-1 Modulation in
HEK293T Cells
[0954] HEK293T cells (ATCC) are seeded in six-well plates and
transfected using Fugene HD to express transiently ACMSD. 24 hrs
post transfection, the cells are stimulated for 48 hrs to 72 hrs
with different concentrations of a compound of Formula (I) or a
pharmaceutically acceptable salt thereof, and then lysed to measure
the ACMSD activity, by measuring the conversion of 30H-Anthranilic
Acid into product (i.e.
.alpha.-amino-.beta.-carboxymuconate-.epsilon.-semialdehyde, ACMS)
in a spectrophotometrical in vitro assay. The amount of the whole
protein content in cell lysates is detected by Bradford analysis.
This value is used to get the specificity activity of the enzyme
normalized in all samples (mU/ml or .DELTA.E/.DELTA.t/mg of total
protein).
[0955] ACMSD-1 enzyme is known to be expressed in liver, kidney and
brain; available cell lines for these cell types were therefore
tested to determine the expression levels of ACMSD. We determine
whether ACMSD-1 is not expressed in transformed cell lines from
liver and kidney, such as HepG2, HEK293T, Hep3B, etc. Transfection
of ACMSD was performed to express the enzyme in different cellular
backgrounds such as COS-7, HEK293T, and HepG2. The HEK293T cellular
background proved to be the best system, with the highest protein
production allowing robust measurement ACMSD1 enzyme activity. This
is probably due to the better transfection efficacy observed in
HEK293T.
[0956] Having determined the optimum stimulation time and
transfection protocol cells are stimulated with different
concentrations of a compound of Formula (I) or a pharmaceutically
acceptable salt thereof (about 50 nM to about 5 uM).
Biological Example 3: Determination of NAD.sup.+ Content in Human
Primary Hepatocytes Treated with a Compound of the Disclosure
[0957] The NAD.sup.+ concentration or content is determined in
human primary hepatocytes treated with a compound of Formula (I) or
a pharmaceutically acceptable salt thereof, Vehicle (NT) was used
as a control.
[0958] At least three experiments are run treating primary
hepatocytes with different concentrations of a compound of Formula
(I) or a pharmaceutically acceptable salt thereof, (0.5 .mu.M and 5
.mu.M) after 48 hrs from seeding. The compounds are replaced every
24 hrs, and then cells are directly harvested and lysed with
ACN/H.sub.2O (ratio 5:1). LCMS/MS is used to detect and measure
NAD.sup.+ concentration/content.
Biological Example 4: Determination of NAD.sup.+ Content in Human
Primary Hepatocytes Treated with a Compound of the Disclosure
[0959] The NAD.sup.+ concentration or content is determined in
human primary hepatocytes treated with a compound of Formula (I) or
a pharmaceutically acceptable salt thereof, and MEHP, a known ACMSD
inhibitor. MEHP is used as a control.
[0960] At least three experiments are run treating primary
hepatocytes with different concentrations of a compound of Formula
(I) or a pharmaceutically acceptable salt thereof, (0.5 .mu.M, 5
.mu.M, and 50 .mu.M) after 48 hrs from seeding. The compounds are
replaced every 24 hrs, and then cells are directly harvested and
lysed with ACN/H.sub.2O (ratio 5:1). LCMS/MS is used to detect and
measure NAD.sup.+ concentration/content.
Biological Example 5: Modulation of SOD2 Activity in AML-12 Cells
and Murine Primary Hepatocytes
[0961] The modulation of SOD-2 activity in AML-12 cells and murine
primary hepatocytes treated with a compound of Formula (I) or a
pharmaceutically acceptable salt thereof, is measured.
[0962] The mouse hepatocytes cell line AML-12 (alpha mouse liver
12) is obtained from ATCC and grown at 37.degree. C. in a
humidified atmosphere of 5% CO.sub.2/95% air in Dulbecco's Modified
Eagle Medium/Nutrient Mixture F-12 (DMEM/F-12) supplemented with
0.005 mg/ml insulin, 0.005 mg/ml transferrin, 5 ng/ml selenium, 40
ng/ml dexamethasone and 1% gentamycin. ACMSD inhibitors are
initially diluted from powder in DMSO to a stock concentration of 1
mM. This stock is further diluted with water to a concentration of
100 .mu.M which was used for the cell treatments.
[0963] Primary hepatocytes are prepared from 8-12-week-old C57BL/6J
mice by collagenase perfusion method. Mouse livers are perfused
with Hank's balanced salt solution (HBSS, KCl, 5.4 mM;
KH.sub.2PO.sub.4, 0.45 mM; NaCl, 138 mM; NaHCO.sub.3, 4.2 mM;
Na.sub.2HPO.sub.4, 0.34 mM; glucose, 5.5 mM; HEPES, 1 M; EGTA, 50
mM; CaCl.sub.2, 50 mM; pH 7.4). Livers are then washed at a rate of
5 ml/min through the portal vein. After washing, livers are
perfused with collagenase (0.025%) solution. Cell viability is
assessed by the trypan blue method. Isolated primary hepatocytes
are plated with DMEM medium (Gibco) including 10% FCS, 10 units per
ml penicillin and HEPES for buffering. The cells are maintained in
culture at 37.degree. C. in a humidified atmosphere of 5%
CO.sub.2/95% air. After 6-8 hrs of attachment, this medium is
replaced with media containing different concentrations of an ACMSD
inhibitor (i.e., compound of Formula (I) or a pharmaceutically
acceptable salt thereof) or with the corresponding concentration of
DMSO (as a control). Primary hepatocytes are harvested about 24 hrs
later if not indicated differently.
[0964] Primary hepatocytes or AML-12 cells are then lysed in a 20
mM HEPES buffer (Gibco), pH 7.2, containing 1 mM EGTA (Sigma), 210
mM mannitol (Sigma), and 70 mM sucrose (AMRESCO). Total protein
concentration is determined using the Bradford assay (BioRad).
SOD-2 activity is determined at various times after ACMSD inhibitor
treatment by the SOD Assay Kit (Cayman Chemical) according to the
manufacturer's instructions. In order to specifically detect the
SOD2 activity 2 mM potassium cyanide is added to the assay, which
inhibits both Cu/Zn-SOD and extracellular SOD, resulting in the
detection of only Mn-SOD (SOD-2) activity. Absorbance is determined
with a Victor X4 multi-label plate reader (Perkin-Elmer) at 450 nm.
Results are expressed in U/ml/mg of protein according to the
standard curve and measured protein concentration.
[0965] The oxidative stress resistance pathway is explored by
measuring the activity of SOD2.
Biological Example 6: Determination of NAD.sup.+ Content in Murine
Primary Hepatocytes
[0966] NAD.sup.+ levels are determined in human primary hepatocytes
treated with a compound of Formula (I) or a pharmaceutically
acceptable salt thereof.
[0967] NAD.sup.+ is extracted using acidic extraction method.
Samples are collected and homogenized in 70% ice-cold perchloric
acid (HClO.sub.4). After insoluble protein parts are pelleted by
adding potassium carbonate (K.sub.2CO.sub.3), the samples are
separated by high-performance liquid chromatography (HPLC) and
analyzed by mass-spectrometry. The proteins in the pellet are
quantified by Bradford assay and were used for normalization.
[0968] The exposure of primary hepatocytes to 5 nM, 10 nM and 50 nM
of an ACMSD inhibitor of Formula (I) or a pharmaceutically
acceptable salt thereof, for 24 hours is examined for significant
and dose-dependent increases in intra-cellular NAD.sup.+
levels.
Biological Example 7: RT-qPCR Analysis of SIRT1-Regulated Genes in
AML-12 Cells, Hepa-1.6 Cells and Primary Murine Hepatocytes Treated
with a Compound of the Disclosure
[0969] Gene expression of ACMSD and genes known to be regulated by
SIRT1, (an enzyme that is strictly NAD.sup.+ dependent) such as
Pgc1a, Sod1, Sod2 (MnSOD), are analyzed in AML-12 cells, Hepa-1.6
cells and primary murine hepatocytes treated with a compound of
Formula (I) or a pharmaceutically acceptable salt thereof.
[0970] Cells (AML-12, Hepa-1.6, HEK-293, primary human and murine
hepatocytes) are treated with different concentrations of a
compound of Formula (I) or a pharmaceutically acceptable salt
thereof. Total RNA is extracted from cells using TRIzol
(Invitrogen) according to the manufacturer's instructions. The RNA
is treated with DNase, and 2 .mu.g of RNA is used for reverse
transcription (RT). 50.times. diluted cDNA is used for
RT-quantitative PCR (RT-qPCR) reactions. The RT-qPCR reactions are
performed using the Light-Cycler system (Roche Applied Science) and
a qPCR Supermix (QIAGEN) with the indicated primers. The average of
at least three technical repeats is used for each biological data
point.
[0971] A dose-dependent increase in mRNA expression levels of genes
is known to be regulated by SIRT1, (an enzyme that is strictly
NAD.sup.+ dependent) such as Pgc1a, Sod2 (MnSOD), but not Sod1
(Cu--Zn SOD). Primary mouse hepatocytes are treated for 24 hrs with
a compound of Formula (I.sub.) or a pharmaceutically acceptable
salt thereof, (5 nM-500 nM range) and are observed for changes in
expression levels and mRNA levels of Pgc1a and Sod2 (MnSOD).
Changes in mRNA expression are compatible with the activation of
SIRT1, subsequent to the induction in NAD.sup.+ levels by
inhibition of ACMSD1 activity.
Biological Example 8: Modulation of Caspase 3/7 Activity in MDCK
Cells
[0972] An in vitro study is performed to determine the effects of
compounds of Formula (I) or a pharmaceutically acceptable salt
thereof, on Acute Kidney Injury in MDCK cells.
[0973] MDCK cells (MDCK (NBL-2) ATCC.RTM. CCL-34.TM.) are cultured
in base medium ATCC-formulated Eagle's Minimum Essential Medium,
Catalog No. 30-2003 with fetal bovine serum (FBS) to a final
concentration of 10%. 10,000 cells are plated into 96 wells and 24
hours after cell plating the medium is changed with fresh medium
supplemented with 1% FBS. Cisplatin (50 .mu.M for 16 hrs) is then
used to induce cell injury. Different concentrations (about 1 .mu.M
to about 125 .mu.M) of a compound of Formula (I) or a
pharmaceutically acceptable salt thereof, (in 1% DMSO) are added in
combination with cisplatin or 1 hour prior adding cisplatin.
[0974] Caspase 3/7 activity (Promega) is determined according to
standard procedures using a luminescent signal readout on a Victor
V plate reader (PerkinElmer). Each experiment/condition is
performed in triplicate.
[0975] Caspase activity is analyzed as percentage effect normalized
to the cisplatin alone (100%) and vehicle treated cells as 0% of
caspase activity. Data are analyzed by GraphPad Software. One-way
analysis of variance (Dunnett's Multiple Comparison test) is used
for statistical analyses.
[0976] MDCK cells are treated with different concentrations of a
compound of Formula (I) or a pharmaceutically acceptable salt
thereof.
Biological Example 9: Cytotoxicity and hERG Screening
[0977] Cytotoxicity: 20000 HePG2 and AML-12 cells are seeded in 96
well plate (Viewplate PerkinElmer). Dose-response of a compound of
Formula (I) or a pharmaceutically acceptable salt thereof, is
performed using HP D300 digital dispenser, ranging from 10 nM to
300 .mu.M with constant DMSO 1% in medium. Cells are stimulated for
4 hrs at 37.degree. C.; the supernatant is used to perform LDH
release (Cytotox-one, Promega) as a measure of necrosis while the
cells are lysed to detect ATP level for determining cell viability
(Celltiter-glo, Promega) according to manufacturer's
instructions.
[0978] The Predictor hERG assay kit (Invitrogen), containing
membrane preparations from Chinese hamster ovary cells stably
transfected with hERG potassium channel and a high-affinity red
fluorescent hERG channel ligand (tracer), is used for the
determination of hERG channel affinity binding of the compounds of
Formula (I) or a pharmaceutically acceptable salt thereof.
Compounds that bind to the hERG channel protein (competitors) are
identified by their ability to displace the tracer, resulting in a
lower fluorescence polarization. The final concentration of DMSO in
each well is maintained at 1%. The assays are performed according
to the manufacturer's protocol (Invitrogen).
Biological Example 10: Anti-Diabetic Effects in C57BL/6J and KK-Ay
Mice
[0979] A glucose tolerance test is performed on male C57BL/6J and
KK-Ay mice to determine the effects of compounds of Formula (I) or
a pharmaceutically acceptable salt thereof, on glucose and insulin
levels.
[0980] Male C57BL/6J and KK-Ay mice, 6-7 weeks of age, are
obtained, e.g., from Charles River Laboratories France and CLEA
Japan, respectively. Mice are fed from the age of 8 weeks onwards
with regular chow (CD-Harlan 2018), a high fat diet (HFD-Harlan
06414). A compound of Formula (I) or a pharmaceutically acceptable
salt thereof, is mixed with the HFD at 180 mg kg.sup.-1 of food. On
the basis of their daily food intake, this results in a daily dose
of about 15 mg kg.sup.-1 body weight. The mice are fasted for 4 hrs
before blood and tissues are harvested for RNA isolation, lipid
measurements and histology. Oxygen consumption is measured with the
Oxymax apparatus (Columbus Instruments). Histological analysis and
transmission electron microscopy are performed.
[0981] An oral glucose tolerance test is performed in the animals
that are fasted overnight. Glucose is administered by gavage at a
dose of 2 g/kg. An intraperitoneal insulin tolerance test is
performed in animals fasted for 4 hrs. Insulin is injected at a
dose of 0.75 U/kg body weight. Glucose is quantified with the Maxi
Kit Glucometer 4 (Bayer Diagnostic) or Glucose RTU (bioMerieux
Inc.) and plasma insulin concentrations are measured by ELISA
(Cristal Chem Inc.). Statistical differences are determined by
either ANOVA or Student's t-test.
Biological Example 11: Anti-Diabetic and Obesity Effects in Db/Db
Mice with LepR Mutation
[0982] A study of the anti-diabetic effects of the compounds of
Formula (I) or a pharmaceutically acceptable salt thereof, is
conducted in genetically obese Leprdb/J (db/db) mice.
[0983] Animals are bred and housed in a temperature- and
humidity-controlled environment in compliance with
FELASA-protocols. From an age of three weeks, mice are fed a
high-fat diet (HFD) (Harlan 06414). Most pharmacological studies
are started in diabetic eight-week-old db/db and wild type (wt)
references.
Subchronic Intervention
[0984] db/db mice are treated once/day with a compound of Formula
(I) or a pharmaceutically acceptable salt thereof, for 14 days
between 5-6 PM before dark-phase onset (6 PM). Blood samples are
collected after 4 hrs of fasting the mice prior to the first dose
and at 18.+-.2 hrs after the last dose. Glucose concentrations of
each blood sample are determined.
Acute Intervention Glucose
[0985] Initial blood samples are collected in random-fed db/db mice
between 6-8 AM after light-phase-onset (6 AM), then compounds of
Formula (I) or a pharmaceutically acceptable salt thereof, are
administered, diet-access is restricted, and the second blood
sample is collected 4 hrs post-treatment. Thereafter, mice are
subjected to an oral glucose tolerance test (OGTT1: 1 g glucose/kg
body mass) and blood glucose concentrations are determined at 0.5,
1, 2, 3, and 4 hrs after each glucose challenge.
Euglycemic-Hyperinsulinemic Clamps Assay
[0986] db/db mice receive a permanent jugular vein catheter under
ketamine/xylazine anesthesia. For six to seven days, later (after 6
AM) food-access is restricted. Conscious mice are placed in
oversized rat-restrainers and warmed by warming pads. Catheter-ends
are then connected to syringes in CMA402-pumps (Axel Semrau,
Sprockhoevel, Germany). After 110 minutes of primed-continuous
[3-.sup.3H]glucose infusion (1.85 kBq/min), a blood sample is
collected to determine plasma insulin, glucose and
[3-.sup.3H]glucose concentrations and to calculate basal endogenous
glucose appearance rates. The mice then receive vehicle or a
compound of Formula (I) or a pharmaceutically acceptable salt
thereof, via gavage.
[0987] Subsequently, glucose-1 clamps are started with a
[3-.sup.3H]glucose infusion (3.7 kBq/min) containing insulin (36
pmol/kg*min.sup.-1; HumulinR, Lilly, USA) causing a moderate
net-increase in plasma insulin concentrations. Blood glucose
concentrations are measured every 10 minutes and target glycemia is
established by adjusting the rate of a 20% glucose infusion (GIR).
At minute 120, 2-deoxy-D-[1-.sup.14C]glucose (370 kBq) is given
intravenously. Blood samples are collected at minute 30, 60, 90,
100, 110, 120, 122, 125, 130, and 140. The mice are then sacrificed
(i.e., through an intravenous ketamine/xylazine-overdose).
Gastrocnemius muscle and epididymal adipose tissue are collected,
immediately snap-frozen in liquid nitrogen, and stored at
-80.degree. C. 2-[.sup.14C]deoxyglucose-6-phosphate is extracted
from the tissue and glucose uptake rates (R.sup.g) are
calculated.
[0988] Plasma [.sup.3H].sup.- and [.sup.14C]-radioactivity is
determined in deproteinized plasma after [.sup.3H.sub.2O]
evaporation. Glucose fluxes under basal conditions and between
glucose clamp minute 60 to 90 and 90 to 120 are estimated as
follows: whole-body glucose disappearance rate (Rd)=[3-.sup.3H]GIR
(dpm/min)/plasma [3-.sup.3H]glucose specific activity
(dpm/min*mol); basal Endo Ra=[3-.sup.3H]GIR (dpm/min)/plasma
[3-.sup.3H]glucose specific activity (dpm/min*mol); glucose-clamp
Endo Ra=GIR-Rd. Ultima-Gold scintillation-cocktail, radioisotopes,
and a Tri-Carb2910TR are obtained from Perkin Elmer (Germany).
Assays from Blood, Plasma, Urine
[0989] Blood samples are collected from lateral tail veins. Blood
glucose is measured with a glucometer (Contour, Bayer Vital,
Germany), urine and plasma glucose with a colorimetric Glucose
LabAssay (Wako, Germany), and HbAlc with AlcNow+(Bayer Vital) or
Clover Analyzer (Inopia, South Korea).
Analyses of Disease Onset and Survival
[0990] Disease onset is defined as the last day of individual peak
body weight before gradual loss occurs. The stages of disease are
defined as follows: the early stage of disease is defined as the
duration of time between peak body weight until loss of 10% of peak
body weight. The late stage of disease is defined as the duration
of time between 10% loss of peak body weight until the end stage of
disease. The end stage of disease is defined as the day when an
animal could no longer right itself within 30 s for three
consecutive trials when placed on its side. Animals are euthanized
at the end stage of disease.
Body Composition Measurements
[0991] Body weights are assessed weekly for at least 13 weeks.
Brown adipose tissue (BAT) and gonadal white adipose tissue (WAT)
are dissected and weighed at the indicated age. Total lean mass, %
of WAT and BMD (bone mineral density) are determined by DEXA
(PIXImus DEXA; GE).
Indirect Calorimetry, Food Intake and Activity
[0992] Animals are initially weighed and acclimated to the test
cage. Volume oxygen (VO.sub.2) and volume carbon dioxide production
(VCO.sub.2) are measured every 20 min using the Oxymax
Comprehensive Laboratory Animal Monitoring System (CLAMS) (Columbus
Instruments) and are reported as average VO.sub.2 per hour
normalized to body weight (mL/h/kg). Using the CLAMS machine,
activity counts by infrared beam interruptions and food intake are
simultaneously measured. More specifically, food intake is measured
by deducting the weight of powderized food pellets at the end of
experimentation from the starting weight at the beginning of
experimentation. To complement this experiment and to control for a
novel environment that may affect feeding behaviour, we also
perform a more `manual` experiment, wherein a set weight of food
pellets is placed at the same time each day into a clean home cage,
which holds a mouse. The next day the weight of the remaining
pellets is recorded and deducted from the starting weight. This
experiment is performed for 14 days straight. The body weight of
each mouse is also recorded daily. Results for each genotype are
similar to that acquired from the CLAMS.
Statistical Analyses.
[0993] Considering a 1-.beta. larger than 0.9 statistically
powerful, we estimate appropriate group numbers from pilot studies
apriori. One- or two-way Analyses of Variance (Bonferroni
post-tests) or t-tests are performed.
Biological Example 12: Effects on Non-Alcoholic Fatty Liver Disease
(NAFLD) and Non-Alcoholic Steatohepatitis (NASH) in Mice
[0994] A study is performed to determine the effects of compounds
of Formula (I) or a pharmaceutically acceptable salt thereof, on
non-alcoholic fatty liver disease (NAFLD) and non-alcoholic
steatohepatitis (NASH) in male C57BL/6J fed a high fat and high
sucrose diet.
[0995] Male C57BL/6J mice (The Jackson Laboratory, Bar Harbor, Me.,
USA) are housed under a 14 hrs light-10 hrs dark cycle at
21-23.degree. C. and have ad libitum access to water during the
entire experiment. From the age of 6 weeks, mice are fed a
`Western` HF-HSD with 44.6% of kcal derived from fat (of which 61%
saturated fatty acids) and 40.6% of kcal derived from carbohydrates
(primarily sucrose 340 g/kg diet) (TD.08811, 45% kcal Fat Diet,
Harlan Laboratories Inc., Madison, Wis., USA) or normal chow diet
(NCD) as control (V1534-000 ssniff R/M-H, ssniff Spezialdiaten
GmbH, Soest, Germany). The animals are then treated with a compound
of Formula (I) or a pharmaceutically acceptable salt thereof, or a
control for 4, 12 or 20 weeks (n=8 per group for every time point),
after which they are sacrificed.
[0996] Body weight and food intake are monitored weekly on the same
day. After sedation with sodium pentobarbital (intraperitoneal
injection, 50 mg/kg body weight), total fat mass is analyzed by
dual-energy X-ray absorptiometry (DEXA) (PIXImus densitometer,
Lunar Corp., Madison, Wis., USA). Intraperitoneal glucose tolerance
test (IPGTT) is performed in 6 hrs fasted mice. Tail vein glucose
levels are measured with a Bayer Contour glucometer immediately
before (time point 0 min) and 15, 30, 60, 90 and 150 min after
glucose administration (1 g glucose/kg body weight). Insulin
resistance is calculated using the Homeostasis Model of Insulin
Resistance (HOMA-IR) index: (fasting insulin (ng/mL).times.fasting
glucose (mg/dL))/405.
Sacrifice
[0997] After a 6 hrs fasting period, mice are anaesthetised with
sodium pentobarbital (intraperitoneal injection, 50 mg/kg body
weight) and sacrificed by blood sampling via cardiac puncture.
Plasma is obtained by centrifugation of blood (6000 rpm for 5 min
at 4.degree. C.) that is collected in heparinised syringes. Tissues
are either snap frozen in liquid nitrogen or stored at -80.degree.
C. together with the plasma until further biochemical and molecular
analyses or preserved for histological analysis.
Histological Analyses
[0998] Liver samples are routinely fixed in buffered formalin (4%)
and embedded in paraffin. Serial 4 mm thick sections are stained
with H&E and picrosirius red to assess fibrosis. Frozen liver
sections are stained with Oil Red O to assess lipid accumulation.
All liver biopsies are analyzed by an expert liver pathologist,
blinded to the dietary condition or surgical intervention.
Steatosis, activity and fibrosis are semiquantitatively scored
according to the NASH-Clinical Research Network criteria. The
amount of steatosis (percentage of hepatocytes containing fat
droplets) is scored as 0 (<5%), 1 (5-33%), 2 (>33-66%) and 3
(>66%). Hepatocyte ballooning is classified as 0 (none), 1 (few)
or 2 (many cells/prominent ballooning). Foci of lobular
inflammation are scored as 0 (no foci), 1 (<2 foci per
200.times. field), 2 (2-4 foci per 200.times. field) and 3 (>4
foci per 200.times. field). Fibrosis is scored as stage F0 (no
fibrosis), stage Fla (mild, zone 3, perisinusoidal fibrosis), stage
F1b (moderate, zone 3, perisinusoidal fibrosis), stage F1c
(portal/periportal fibrosis), stage F2 (perisinusoidal and
portal/periportal fibrosis), stage F3 (bridging fibrosis) and stage
F4 (cirrhosis). Diagnosis of NASH is based on accepted histological
criteria. Severity of the disease is assessed using the NAS (NAFLD
activity score) as the unweighted sum of scores of steatosis,
hepatocyte ballooning and lobular inflammation. Percentage of
fibrosis is quantitated by morphometry from digitalised sirius red
stained sections using the Aperio system after tuning the threshold
of fibrosis detection under visual control. Results are expressed
as collagen proportional area.
Biological Example 13: Effects on Non-Alcoholic Fatty Liver Disease
(NAFLD) and Non-Alcoholic Steatohepatitis (NASH) in Methionine and
Choline Deficient Mice
[0999] A study is performed to determine the effects of compounds
of Formula (I) or a pharmaceutically acceptable salt thereof, on
non-alcoholic fatty liver disease (NAFLD) and non-alcoholic
steatohepatitis (NASH) in male wildtype mice fed a methionine- and
choline-deficient diet.
[1000] Wildtype mice housed in 12-hour light/dark cycles, with free
access to food and water are used. At least 5 animals per time
point are analyzed. All experiments are repeated at least three
times. For dietary treatment, 8-12 weeks old male mice weighing 25
g are either fed a methionine- and choline-deficient diet (MCD to
induce NASH) or chow diet (as a control). Animal experiments and
evaluation of NAFLD and NASH as described above in Example 55 for
mice fed the high fat and high sucrose diet.
Biological Example 14: Effects on Atherosclerosis in High
Cholesterol Fed LDL-R Knockout Mice
[1001] A study is performed to determine the effects of compounds
of Formula (I) or a pharmaceutically acceptable salt thereof, on
atherosclerosis in high cholesterol fed LDL-R knockout mice.
[1002] LDL-R knockout (KO) mice are backcrossed for ten generations
with the C57BL/6J strain, yielding congenic C57BL/6J animals. The
controls that are used are littermates in all experiments. The
animals are treated with a compound of Formula (I) or
pharmaceutically acceptable salt thereof, or a control. Mice are
sacrificed 12 weeks after the initiation of the atherogenic diet
(TD94059; Harlan), after which the heart and aorta are perfused
with PBS and subsequently fixed (Shandon Formal Fixx, Thermo
Scientific). Atherosclerosis is assessed by an Oil red 0 staining
of the aortic root and quantified with MetaMorph software.
Biochemistry parameters are measured with the appropriate kits in
the COBAS C111 (Roche). For the in vivo lipopolysaccharide (LPS)
study, mice are intraperitoneally injected with 100 mg of LPS, and
blood is taken from the tail vein. TNF.alpha. levels are quantified
with Mouse TNF.alpha. ELISA Ready-SET-Go! (eBioscience) assay.
Blood cell counts are determined with Advia2120 (Siemens Healthcare
Diagnostics).
[1003] The Student's t test is used to calculate the statistical
significance. In case of multiple testing (i.e., the comparison of
more than two groups), this test is preceded by the ANOVA test.
P<0.05 is considered statistically significant. Results
represent the mean SEM.
Biological Example 15: Effects on Inherited Mitochondrial Disease
in Sco2.sup.KO/KI Mice
[1004] A study is performed to determine the effects of compounds
of Formula (I) or a pharmaceutically acceptable salt thereof, on
inherited mitochondrial disease in Sco2.sup.KO/KI mice.
[1005] Anti-COI, anti-COX5a, anti-Ndufa9, anti-SDH-HA, and
anti-Core 2 are from Invitrogen; anti-GAPDH is from Millipore;
anti-FoxO1 and anti-acetylated-FoxO1 are from Cell Signaling and
Santa Cruz, respectively. Anti-mouse secondary antibodies are from
Amersham. Chemicals are from Sigma. Oligonucleotides are from
PRIMM, Italy.
[1006] Compounds of Formula (I) or a pharmaceutically acceptable
salt thereof, are dissolved in water and added to a standard powder
diet (Mucedola, Italy) at the appropriate concentration of 50
mg/Kg/day. Pellets containing the compounds of Formula (I) or a
pharmaceutically acceptable salt thereof, or the vehicles are
reconstituted by hand and kept frozen at -20.degree. C. until
needed. The diet supply is changed every three days, and only the
amount needed is thawed at each time and administered ad libitum
for one month. Sco2.sup.KO/KI mice are maintained in a temperature-
and humidity-controlled animal-care facility, with a 12 hrs
light/dark cycle and free access to water and food. Animals are
sacrificed by cervical dislocation.
Morphological Analysis
[1007] For histochemical analysis, tissues are frozen in
liquid-nitrogen precooled isopentane. Series of 8 mm thick sections
are stained for COX and SDH.
Biochemical Analysis of MRC Complexes
[1008] Muscle quadriceps samples stored in liquid nitrogen are
homogenized in 10 mM phosphate buffer (pH 7.4), and the
spectrophotometric activity of cI, cII, cIII, and cIV, as well as
CS, is measured as described. Note that in all panels the activity
of cII is multiplied by 10 for visualization clarity.
NAD.sup.+ Determination
[1009] NAD.sup.+ is extracted using acidic and alkaline extraction
methods, respectively. Tissue NAD.sup.+ is analyzed with mass
spectrometry as previously described.
Biological Example 16: Effects on Inherited Mitochondrial Disease
in Deletor Mice
[1010] A study is performed to determine the effects of compounds
of Formula (I) or a pharmaceutically acceptable salt thereof, on
inherited mitochondrial disease in Deletor mice.
[1011] The Deletor mouse model is generated in C57BL/6 congenic
background and has been previously characterized (Tyynismaa et al,
2005); WT mice are littermates from the same congenic mouse strain
C57BL/6J. Deletor and WT male mice are administered either chow
diet (CD) or a compound of Formula (I) or a pharmaceutically
acceptable salt thereof, admixed with the CD at the appropriate
concentration. The food pellets are manually prepared by mixing a
compound of Formula (I) or a pharmaceutically acceptable salt
thereof, into the powdered food as described for the Sco2.sup.KO/KI
mice in Example 58 and stored at -20.degree. C. The mice are housed
in standard animal facility, under a 12 hrs dark/light cycle. They
have ad libitum access to food and water. The pre-manifestation
group consists of 12 Deletors and 12 WT mice, and the
post-manifestation group of 24 Deletors and 24 WT mice, receiving
either a compound of Formula (I) or a pharmaceutically acceptable
salt thereof, or CD diet. During the intervention, the mice are
regularly monitored for weight, food consumption, and physical
endurance. Their exercise capability is measured twice by treadmill
exercise test (Exer-6M Treadmill, Columbus Instruments) at the
start and the end of the diet. The exercise test protocol consists
of the initial running speed of 7 m/s which is increased every 2
min by 2 m/s and continued until the animal is unable to run or
repeatedly falls from the belt at the stimulus site.
[1012] Oxygen consumption and carbon dioxide production, as well as
spontaneous moving and feeding activities, are recorded by Oxymax
Lab Animal Monitoring System (CLAMS; Columbus Instruments, OH,
USA). The mice are kept in individual cages inside a CLAMS chamber
for 3 days; the first day and night is a nonrecording adjustment
period followed by a 24 hrs recording at thermoneutrality
(+30.degree. C.). The results of O.sub.2 consumption and CO.sub.2
production are used to calculate respiratory exchange rate and
analyzed separately from the light (inactive) and dark (active)
periods of the day.
Morphologic Analysis
[1013] Tissue sections are prepared from the quadriceps, liver, and
BAT. Samples are embedded with OCT Compound Embedding Medium
(Tissue-Tek) and snap-frozen in 2-methylbutane in liquid nitrogen.
Frozen sections (12 lm) from quadriceps are assayed for in situ
histochemical COX and succinate dehydrogenase (SDH) activities
simultaneously. The activities from the quadriceps sections, the
COX-negative and the COX-negative plus SDH positive and normal
fibres are calculated. Approximately 2000 fibres are counted from
each mouse sample. The intensity of COX histochemical activity from
quadriceps for both oxidative and non-oxidative fibres is measured
with Image J software. Frozen sections (8 .mu.m) from liver and BAT
are stained with Oil Red 0. For plastic embedding, quadriceps,
liver, and BAT samples are fixed in 2.5% glutaraldehyde, treated
with 1% osmium tetroxide, dehydrated in ethanol, and embedded in
epoxy resin. Semi-thin (1 .mu.m) sections are stained with methyl
blue (0.5% w/v) and boric acid (1% w/v). The interesting areas for
the ultrastructural analyses are selected by inspection of the
light microscopic sections. For transmission electron microscopy,
ultrathin (60-90 nm) sections are cut on grids and stained with
uranyl acetate and lead citrate and viewed with a Transmission
Electron Microscope. Crista content in both BAT and muscle is
determined from electron micrographs, utilizing a 1 .mu.m
"intra-mitochondrial measuring stick," placed perpendicular to
cristae. Skeletal muscle samples are also analyzed for citrate
synthase activity.
Biological Example 17: Effects on Kidney Disease
[1014] A study is performed to determine the effects of compounds
of Formula (I) or a pharmaceutically acceptable salt thereof, on
kidney disease in C57BL/6J WT mice. (Wei, Q., et al., "Mouse model
of ischemic acute kidney injury: technical notes and tricks"
American Journal of Physiology-Renal Physiology, 303(11),
F1487-F1494)
[1015] C57BL/6J WT mice are purchased from Charles-River. All mice
are fed a standard commercial diet while housed at an ambient
temperature of 20-22.degree. C. with a relative humidity of
50.+-.5% under 12/12 hrs light-dark cycle in a specific
pathogen-free facility. The experimental mice are 8 weeks old and
are divided into four groups: (1) control (n=5); (2) cisplatin (20
mg/kg; Sigma Chemical, St Louis, Mo.; n=5); (3) a compound of
Formula (I) or a pharmaceutically acceptable salt thereof, and
cisplatin (n=5); and (4) a compound of Formula (I) or a
pharmaceutically acceptable salt thereof, alone (40 mg/kg; n=5).
The dose and time of cisplatin treatment for nephrotoxicity are
chosen according to a published method. A compound of Formula (I)
or a pharmaceutically acceptable salt thereof, is administered
orally once a day for 4 days. Cisplatin is injected once at 12 hrs
after the first administration of a compound of Formula (I) or a
pharmaceutically acceptable salt thereof. The mice are sacrificed
at 72 hrs after the single cisplatin injection.
Assays for Renal Functional Markers and Proinflammatory
Cytokines
[1016] For renal function analysis, serum is isolated and stored at
-80.degree. C. until use. Serum creatinine and BUN levels are
measured using an assay kit according to the manufacturer's
instructions (BioVision, Milpitas, Calif.). In addition, the
proinflammatory cytokines TNF-.alpha., IL-1b, and IL-6 from serum
or homogenates from kidney tissue are quantified by ELISA
(Quantikine Kit; R&D Systems, Minneapolis, Minn.) according to
the manufacturer's instructions. For measuring cytokines, kidney
tissue is homogenized in phosphate buffered saline containing 0.05%
Tween-20. Aliquots containing 300 mg of total protein are used. A
metabolic cage is used for collecting urine to analyze the level of
urinary cytokines. The sample size for each group is five.
Alternative Study of the Effects on Kidney Disease
[1017] Alternatively, C57BL/6J WT mice are numbered and kept in
acclimatization for a period of 5-7 days before initiation of the
experiment. (Wei, Q., et al. "Mouse model of ischemic acute kidney
injury: technical notes and tricks" American Journal of
Physiology-Renal Physiology, 303(11), F1487-F1494) Mice are
randomized into different treatment groups based on their body
weight. Different groups are maintained on Harlan diet 2916. Mice
are then maintained on the respective diets for 10 days prior to
bilateral Ischemic kidney injury. Body weight measurement is made
once at randomization and once on day 7. Food consumption is
evaluated once on day 7. Blood is collected by retro-orbital
puncture under mild Isoflurane anesthesia and used for analysis of
basal blood urea nitrogen levels (BUN) on day 9.
[1018] Mice are anesthetized with ketamine (80 mg/kg i.p) and/or
Xylazine (10 mg/kg, i.p.) and placed on a surgical platform in a
dorsal position. Both kidneys are exposed through flank incisions
and renal pedicles are occluded using vascular clamps for 25
minutes. The clamp is then removed and the surgical site is
sutured. 1 ml of physiological saline is administered
intra-peritoneally after closing the wound to prevent dehydration.
The sham-operated group is subjected to similar surgical
procedures, except that the occluding clamp is not applied. Animals
are monitored until recovery from anesthesia and returned to their
home cage. Animals are observed every day for general clinical
signs and symptoms and mortality.
[1019] One day prior to termination, animals are individually
housed in metabolic cages for 12h and urine is collected for
estimation of urea, creatinine, sodium and potassium.
[1020] On days 12, 14, & 16 blood is collected by retro orbital
puncture under mild isoflurane anesthesia and plasma is used for
analysis of blood urea nitrogen levels (BUN) and serum creatinine.
Animals are then euthanized by CO.sub.2 inhalation and organs are
collected. One kidney is fixed in 10% neutral buffered formalin and
the other is flash frozen in liquid nitrogen, stored at -80.degree.
C. and used for the estimation of lipid peroxidation, GSH, KIM1,
MPO, NGAL, and SOD levels.
Histological Analysis and Neutrophil Counting
[1021] Mouse kidneys are fixed in 4% formaldehyde and embedded in
paraffin wax. The 5-mm-thick sections are deparaffinised in xylene
and rehydrated through graded concentrations of ethanol. H&E
and PAS staining are performed using standard protocols. Images are
collected and analyzed using a light microscope (IX71, Olympus,
Tokyo, Japan) with DP analyzer software (DP70-BSW, Tokyo, Japan).
Tubular damage in PAS-stained kidney sections is examined under a
light microscope and scored based on the percentage of cortical
tubular necrosis: 0=normal, 1=1-10, 2=11-25, 3=26-45, 4=46-75, and
5=76-100%. Slides are scored in a blinded manner, and results are
means.+-.s.d. of 10 representative fields/group. Severity criterion
for tubular necrosis displaying the loss of the proximal tubular
brush border and cast formation are used to classify samples. The
sample size for each group is 10. Neutrophil infiltration is
quantitatively assessed on PAS stained tissue by a renal
pathologist by counting the number of neutrophils per high-power
field (.times.400). At least 10 fields are counted in the outer
stripe of the outer medulla for each slide.
[1022] All values are represented as mean.+-.s.d. One-way analysis
of variance is used to calculate the statistical significance of
the results of all assays and P-values <0.05 are considered
statistically significant.
Biological Example 18: Effects on Ischemia/Reperfusion-Induced
Acute Kidney Injury
[1023] A study is performed to determine the effects of compounds
of Formula (I) or a pharmaceutically acceptable salt thereof, on
Ischemia/Reperfusion-induced (I/R-induced) Acute Kidney Injury in
CD-1 (ICR) mice.
[1024] CD-1 (ICR) mice are purchased from Charles River Laboratory
(Wilmington, Mass.). Mice are housed in a temperature- and
humidity-controlled environment with a 12:12 hrs light-dark cycle
and are allowed freely access to standard rodent chow (TekLad,
Madison, Wis.) and tap water.
[1025] Mice are subjected to a midline back incision, and both
renal pedicles are clamped for 45 min with microaneurysm clamps
(00396-01; Fine Science Tools, Foster City, Calif.). After removal
of the clamp, the kidneys are inspected for the restoration of
blood flow. The animals are allowed to recover, and they are
sacrificed 48 hrs after reperfusion. Mice are treated with 100
mg/kg of a compound of Formula (I) or a pharmaceutically acceptable
salt thereof, by oral gavage once per day. CD-1 mice are divided
into four groups: (1) young mice with sham injury (n=4) (6-7 weeks
old); (2) young mice with I/R injury (n=8); (3) adult mice with
sham injury (n=4) (20-24 weeks old); and (4) adult mice with I/R
injury (n=11). An additional 27 adult mice (20-24 weeks old) are
randomized into two groups: 13 mice received a compound of Formula
(I) or a pharmaceutically acceptable salt thereof, and the other 14
mice received the vehicle as a control.
[1026] The serum creatinine level is measured using the QuantiChrom
Creatinine Assay Kit (DICT-500, BioAssay Systems, Hayward, Calif.).
BUN measurements are recorded using the Infinity Urea (Nitrogen)
Liquid Stable Reagent (TR12421; ThermoTrace, Victoria, AU).
Evaluation of Renal Tissue
[1027] Kidneys are fixed in 4% paraformaldehyde, embedded in
paraffin, and stained with hematoxylin and eosin (4 mm thick).
Tubular injury is scored on a scale of 0-4 on the basis of the
percentage of tubules with necrosis, dilatation, or cell swelling:
0, less than 5%; 1, 5-25%; 2, 25-50%; 3, 50-75%; and 4, over 75%.
All high-power fields (.times.400) in the cortex and outer medulla
are evaluated by a pathologist in a blinded manner.
[1028] All values are expressed as mean.+-.s.e. Statistical
analysis is carried out using GraphPad Prism 4.00 (San Diego,
Calif.) with unpaired Student's t testing for two sets of data and
an analysis of variance with a Bonferroni post-test for multiple
groups. P<0.05 was considered significant.
Biological Example 19: Effects on Cisplatin-Induced Acute Kidney
Injury
[1029] A study is performed to determine the effects of compounds
of Formula (I) or a pharmaceutically acceptable salt thereof, on
Cisplatin-induced Acute Kidney Injury in C57BL6 mice.
[1030] C57BL6 mice are purchased from Charles River Laboratory
(Wilmington, Mass.). Mice are housed in a temperature- and
humidity-controlled environment with a 12:12 hrs light-dark cycle
and are allowed freely access to standard rodent chow (TekLad,
Madison, Wis.) and tap water.
[1031] Mice are subjected to a midline back incision, and both
renal pedicles are clamped for 45 min with microaneurysm clamps
(00396-01; Fine Science Tools, Foster City, Calif.).
[1032] Cisplatin was injected intraperitoneally 20 mg/kg body
weight. The animals are allowed to recover, and they are sacrificed
48,72, and 96 hrs post cisplatin injection. Mice are treated with
100 mg/kg of a compound of Formula (I) or a pharmaceutically
acceptable salt thereof, by oral gavage once per day. C57BL6 mice
are divided into four groups: (1) young mice with sham injury (n=4)
(6-7 weeks old); (2) young mice with Cisplatin injury (n=8); (3)
adult mice with sham injury (n=4) (20-24 weeks old); and (4) adult
mice with Cisplatin injury (n=11). An additional 27 adult mice
(20-24 weeks old) are randomized into two groups: 13 mice received
a compound of Formula (I) or a pharmaceutically acceptable salt
thereof, and the other 14 mice received the vehicle as a
control.
[1033] The serum creatinine level is measured using the QuantiChrom
Creatinine Assay Kit (DICT-500, BioAssay Systems, Hayward, Calif.).
BUN measurements are recorded using the Infinity Urea (Nitrogen)
Liquid Stable Reagent (TR12421; ThermoTrace, Victoria, AU).
Evaluation of Renal Tissue
[1034] Kidneys are fixed in 4% paraformaldehyde, embedded in
paraffin, and stained with hematoxylin and eosin (4 mm thick).
Tubular injury is scored on a scale of 0-4 on the basis of the
percentage of tubules with necrosis, dilatation, or cell swelling:
0, less than 5%; 1, 5-25%; 2, 25-50%; 3, 50-75%; and 4, over 75%.
All high-power fields (.times.400) in the cortex and outer medulla
are evaluated by a pathologist in a blinded manner.
[1035] All values are expressed as mean.+-.s.e. Statistical
analysis is carried out using GraphPad Prism 4.00 (San Diego,
Calif.) with unpaired Student's t testing for two sets of data and
an analysis of variance with a Bonferroni post-test for multiple
groups. P<0.05 was considered significant.
Biological Example 20: Effects on Sepsis-Induced Acute Kidney
Injury
[1036] A study is performed to determine the effects of compounds
of Formula (I) or a pharmaceutically acceptable salt thereof,
against sepsis-induced Acute Kidney Injury in C57BL6 mice (12-15
weeks old).
[1037] C57BL6 mice are purchased from Charles River Laboratory
(Wilmington, Mass.). Mice are housed in a temperature- and
humidity-controlled environment with a 12:12 hrs light-dark cycle
and are allowed freely access to standard rodent chow (TekLad,
Madison, Wis.) and tap water. Sepsis is induced by Cecal ligation
and puncture (CLP). The procedure for CLP is as follows; the lower
abdomen is shaved, and cleaned with 70% ethanol and a 1 cm
laparotomy is performed whereby the cecum is identified and
externalized. The cecum is measured from the ileo-cecal valve to
the tip, ligated at the .about.50% mark, punctured once
(through-and-through) with a 21-gauge needle and returned into the
abdomen after expressing to allow for fecal matter to extrude. The
abdominal wall is then closed by planes using a running Silk 4-0.
Control animals undergo the same laparotomy, identification and
externalization of the cecum, but no ligation or perforation.
Animals are then given 1 ml of saline and buprenorphine into the
scruff of the neck subcutaneously and recovered on thermal blankets
under monitoring. Animals are treated with ampicillin sulbactam
(250 mg/kg Q12 hours IP for 3 days) and analgesic treatment
(buprenorphine 0.05 mg/kg for 3 days).
[1038] A compound of Formula (I) or a pharmaceutically acceptable
salt thereof, are administered at two different timepoints: A. At
the time of CLP (early phase); and B. At 24 hours after CLP via IP
injection.
[1039] Blood and kidney tissues are collected at the time of
sacrificing for measurement of primary and secondary endpoints.
[1040] Primary endpoint (at 48 hrs):serum creatinine. Secondary
endpoints (at 48 hours) include: Macrophage phenotype marker (IF
stain), Plasma NGAL, Plasma and kidney markers of inflammation
(IL-6, IL-18, TNF), and Kidney Injury markers (KIM-1, NGAL, TIMP2
and IGFBP7). Addition endpoints include; cell death (IF: Annexin V
and Propidium Iodide; Caspase 3/7), autophagy, biogenesis
(PGC-1.alpha., mitochondrial DNA), OXPHOS (Complex I, III, IV
activity), Sirt1 and Sirt3 expression, AMPK (Total, P-AMPK, P-ACC,
and HIF-1.alpha..
Histological Analysis
[1041] Mouse kidneys are fixed in 4% formaldehyde and embedded in
paraffin wax. The 5-mm-thick sections are deparaffinised in xylene
and rehydrated through graded concentrations of ethanol. H&E
and PAS staining are performed using standard protocols. Images are
collected and analyzed using a light microscope (IX71, Olympus,
Tokyo, Japan) with DP analyzer software (DP70-BSW, Tokyo, Japan).
Tubular damage in PAS-stained kidney sections is examined under a
light microscope and scored based on the percentage of cortical
tubular necrosis: 0=normal, 1=1-10, 2=11-25, 3=26-45, 4=46-75, and
5=76-100%. Slides are scored in a blinded manner, and results are
means.+-.s.d. of 10 representative fields/group. Severity criterion
for tubular necrosis displaying the loss of the proximal tubular
brush border and cast formation are used to classify samples. The
sample size for each group is (n=6 per group in groups 2, 4-9 and
n=3 for group 1, 45 total). A tubular injury score will be used to
evaluate protection against kidney damage.
Biological Example 21: Effects on FoxO1 Phosphorylation Levels
[1042] AML-12 cells are treated with different concentrations of a
compound of Formula (I) or a pharmaceutically acceptable salt
thereof, for 24 hours. Cells are then lysed in lysis buffer (50 mM
Tris, 150 mM KCl, EDTA 1 mM, NP40 1%) containing protease and
phosphatase inhibitors, and analyzed by SDS-PAGE/western blot.
Blocking and antibody incubations were done in 5% milk. Each
protein present is detected with its specific antibody. Tubulin
antibody is obtained from Sigma Inc, FoxO1 and phospho-FoxO1
(Ser256) antibodies were obtained from Cell Signaling. Antibody
detection reactions are developed by enhanced chemiluminescence
(Advansta, CA, USA) using X-ray films.
[1043] Unless otherwise defined, all technical and scientific terms
used herein have the same meaning as commonly understood by one of
ordinary skill in the art to which this disclosure belongs. In the
specification, the singular forms also include the plural unless
the context clearly dictates otherwise. Although methods and
materials similar or equivalent to those described herein can be
used in the practice of testing the present disclosure, suitable
methods and materials are described below. All publications, patent
applications, patents, and other references mentioned herein are
hereby expressly incorporated by reference. The references cited
herein are not admitted to be prior art of the claimed disclosure.
In the case of conflict, the present specification, including
definitions, will control. In addition, the materials, methods, and
examples are illustrative only and are not intended to be
limiting.
EQUIVALENTS
[1044] Those skilled in the art will recognize, or be able to
ascertain using no more than routine experimentation, many
equivalents to the specific embodiments and methods described
herein. Such equivalents are intended to be encompassed by the
scope of the present disclosure.
* * * * *