U.S. patent application number 16/825755 was filed with the patent office on 2021-02-04 for novel glutaminase inhibitors.
The applicant listed for this patent is Rhizen Pharmaceuticals SA. Invention is credited to Govindarajulu Babu, Prashant K Bhavar, Gayatri Swaroop Merikapudi, Swaroop K. Vakkalanka, Srikant Viswanadha.
Application Number | 20210032236 16/825755 |
Document ID | / |
Family ID | 1000005153875 |
Filed Date | 2021-02-04 |
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United States Patent
Application |
20210032236 |
Kind Code |
A1 |
Bhavar; Prashant K ; et
al. |
February 4, 2021 |
NOVEL GLUTAMINASE INHIBITORS
Abstract
The present disclosure provides compounds of formula (I) to
(III) as glutaminase inhibitors, methods of preparing them,
pharmaceutical compositions containing them and methods of
treatment, prevention and/or amelioration of diseases or disorders
involving glutamine.
Inventors: |
Bhavar; Prashant K;
(Hyuderabad, IN) ; Vakkalanka; Swaroop K.; (La
Chaux-de-Fonds, CH) ; Viswanadha; Srikant;
(Hyderabad, IN) ; Merikapudi; Gayatri Swaroop;
(Hyderabad, IN) ; Babu; Govindarajulu; (Hyderabad,
IN) |
|
Applicant: |
Name |
City |
State |
Country |
Type |
Rhizen Pharmaceuticals SA |
La Chaux-de-Fonds |
|
CH |
|
|
Family ID: |
1000005153875 |
Appl. No.: |
16/825755 |
Filed: |
March 20, 2020 |
Related U.S. Patent Documents
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Application
Number |
Filing Date |
Patent Number |
|
|
15693291 |
Aug 31, 2017 |
10611759 |
|
|
16825755 |
|
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|
14769331 |
Aug 20, 2015 |
9783533 |
|
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PCT/IB2015/050075 |
Jan 5, 2015 |
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15693291 |
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Current U.S.
Class: |
1/1 |
Current CPC
Class: |
C07D 239/42 20130101;
A61K 31/444 20130101; A61K 31/497 20130101; C07D 241/20 20130101;
A61K 31/506 20130101; A61K 31/4965 20130101; C07D 401/04 20130101;
A61K 31/501 20130101; C07D 401/12 20130101; A61K 31/4545 20130101;
A61K 45/06 20130101; C07D 417/14 20130101; A61K 31/44 20130101;
A61K 31/433 20130101; C07D 417/08 20130101; C07D 285/135 20130101;
A61K 31/505 20130101; Y02A 50/30 20180101; C07D 213/75 20130101;
C07D 417/06 20130101; A61K 31/4439 20130101 |
International
Class: |
C07D 417/14 20060101
C07D417/14; C07D 417/06 20060101 C07D417/06; C07D 285/135 20060101
C07D285/135; A61K 31/433 20060101 A61K031/433; A61K 31/44 20060101
A61K031/44; A61K 31/4439 20060101 A61K031/4439; A61K 31/444
20060101 A61K031/444; A61K 31/4965 20060101 A61K031/4965; A61K
31/497 20060101 A61K031/497; A61K 31/501 20060101 A61K031/501; A61K
31/505 20060101 A61K031/505; A61K 31/506 20060101 A61K031/506; A61K
45/06 20060101 A61K045/06; C07D 213/75 20060101 C07D213/75; C07D
239/42 20060101 C07D239/42; C07D 241/20 20060101 C07D241/20; C07D
401/12 20060101 C07D401/12; C07D 417/08 20060101 C07D417/08; A61K
31/4545 20060101 A61K031/4545; C07D 401/04 20060101 C07D401/04 |
Foreign Application Data
Date |
Code |
Application Number |
Jan 6, 2014 |
IN |
36/CHE/2014 |
Jan 6, 2014 |
IN |
39/CHE/2014 |
May 29, 2014 |
IN |
2639/CHE/2014 |
May 29, 2014 |
IN |
2647/CHE/2014 |
Jun 6, 2014 |
IN |
2783/CHE/2014 |
Jul 18, 2014 |
IN |
3525/CHE/2014 |
Jul 24, 2014 |
IN |
3612/CHE/2014 |
Jul 24, 2014 |
IN |
3613/CHE/2014 |
Oct 31, 2014 |
IN |
5438/CHE/2014 |
Claims
1. A compound of formula (I) ##STR00136## or a tautomer thereof,
prodrug thereof, N-oxide thereof, stereoisomer thereof,
pharmaceutically acceptable ester thereof or pharmaceutically
acceptable salt thereof, wherein L is -L.sub.1-L.sub.2-L.sub.3-;
L.sub.1 is absent or independently selected from substituted or
unsubstituted C.sub.1-6 alkyl, substituted or unsubstituted
C.sub.2-6 alkenyl, substituted or unsubstituted C.sub.2-6 alkynyl,
O, S, --S(.dbd.O).sub.q--, --C(.dbd.O)-- or --NR.sup.x--; L.sub.2
is substituted or unsubstituted 3 to 14 membered heterocyclyl;
L.sub.3 is absent or independently selected from substituted or
unsubstituted C.sub.1-6 alkyl, substituted or unsubstituted
C.sub.2-6 alkenyl, substituted or unsubstituted C.sub.2-6 alkynyl,
O, S, --S(.dbd.O).sub.q--, --C(.dbd.O)-- or --NR.sup.x--; A and B
are independently selected from ##STR00137## wherein each of A and
B are optionally substituted with one or more R.sup.3; each
occurrence of R.sup.3 is, independently hydrogen, halogen,
substituted or unsubstituted C.sub.1-3 alkyl, nitro, amino,
substituted or unsubstituted C.sub.1-6 alkoxy, or substituted or
unsubstituted C.sub.1-6 alkylamino; P and Q are independently
selected from substituted or unsubstituted alkyl, substituted or
unsubstituted alkenyl, substituted or unsubstituted alkynyl,
substituted or unsubstituted alkoxy,
--NR.sup.x--C(.dbd.O)--(CR.sup.xR.sup.y).sub.r--,
--NH--C(.dbd.O)--C(R.sup.xR.sup.y)--,
--(CR.sup.xR.sup.y).sub.r--C(.dbd.O)--NR.sup.x--,
--(CR.sup.xR.sup.y)--C(.dbd.O)--NH--,
--C(.dbd.O)NR.sup.x--(CR.sup.xR.sup.y).sub.r--,
--C(.dbd.O)NH--C(R.sup.xR.sup.y)--,
--(CR.sup.xR.sup.y).sub.r--NR.sup.x--C(.dbd.O)--,
--(CR.sup.xR.sup.y)--NH--C(.dbd.O)--, --NR.sup.x--,
--NR.sup.xC(.dbd.O)--, --NR.sup.xC(.dbd.S)--,
--NR.sup.xS(.dbd.O).sub.q--, --C(.dbd.O)NR.sup.x--,
--C(.dbd.S)NR.sup.x--, --S(.dbd.O).sup.qNR.sup.x--,
--NR.sup.xC(.dbd.O)NR.sup.x--, --NR.sup.xC(.dbd.S)NR.sup.x--,
--C(.dbd.O)--, --C(.dbd.S)--, --C(.dbd.O)ONR.sup.x--,
.dbd.N--N(R.sup.x)--, --N(R.sup.x)--N.dbd. or
--NR.sup.xC(.dbd.O)O--; R.sup.1 and R.sup.2 are independently
selected from hydrogen, hydroxy, halogen, substituted or
unsubstituted alkyl, substituted or unsubstituted alkoxy,
substituted or unsubstituted alkenyl, substituted or unsubstituted
alkynyl, substituted or unsubstituted cycloalkyl, substituted or
unsubstituted cycloalkenyl, substituted or unsubstituted
cycloalkylalkyl, substituted or unsubstituted cycloalkenylalkyl,
substituted or unsubstituted heterocyclyl, substituted or
unsubstituted heterocyclylalkyl, substituted or unsubstituted aryl,
substituted or unsubstituted arylalkyl, substituted or
unsubstituted heteroaryl, substituted or unsubstituted
heteroarylalkyl, --C(.dbd.O)OR.sup.z, --C(.dbd.O)R.sup.z,
--C(.dbd.S)R.sup.z, --C(.dbd.O)NR.sup.zR.sup.z,
--C(.dbd.O)ONR.sup.zR.sup.z, --NR.sup.zR.sup.z,
--NR.sup.zC(.dbd.O)NR.sup.zR.sup.z, --NR.sup.zS(.dbd.O)R.sup.z--,
--NR.sup.zS(.dbd.O).sub.2R.sup.z, --(.dbd.N--N(R.sup.z)R.sup.z),
--NR.sup.zC(.dbd.O)OR.sup.z, --NR.sup.zC(.dbd.O)R.sup.z,
--NR.sup.xC(.dbd.S)R.sup.y, --NR.sup.zC(.dbd.S)NR.sup.zR.sup.z,
--SONR.sup.zR.sup.z, --SO.sub.2NR.sup.zR.sup.z, --OR.sup.z,
--OR.sup.zC(.dbd.O)NR.sup.zR.sup.z, --OR.sup.zC(.dbd.O)OR.sup.z,
--OC(.dbd.O)R.sup.z, --OC(.dbd.O)NR.sup.zR.sup.z,
--R.sup.zNR.sup.zC(.dbd.O)R.sup.z, --R.sup.zOR.sup.z,
--R.sup.zC(.dbd.O)OR.sup.z, --R.sup.zC(.dbd.O)NR.sup.zR.sup.z,
--R.sup.zC(.dbd.O)R.sup.z, --R.sup.zOC(.dbd.O)R.sup.z, --SR.sup.z,
--SOR.sup.z, --SO.sub.2R.sup.z, --CR.sup.xR.sup.yC(.dbd.O)R.sup.z
or --CR.sup.xR.sup.yC(.dbd.S)R.sup.z; each occurrence of R.sup.x,
R.sup.y and R.sup.z is independently selected from hydrogen,
substituted or unsubstituted alkyl, substituted or unsubstituted
alkoxy, substituted or unsubstituted alkenyl, substituted or
unsubstituted alkynyl, substituted or unsubstituted aryl,
substituted or unsubstituted arylalkyl, substituted or
unsubstituted heteroaryl, substituted or unsubstituted
heteroarylalkyl, substituted or unsubstituted cycloalkyl,
substituted or unsubstituted cycloalkylalkyl, substituted or
unsubstituted cycloalkenyl, substituted or unsubstituted
heterocyclic ring, substituted heterocyclylalkyl ring, or
substituted or unsubstituted amino, or any two of R.sup.x and
R.sup.y when bound to a common atom may be joined to form (i) a
substituted or unsubstituted saturated or unsaturated 3-14 membered
ring, which may optionally include one or more heteroatoms which
may be the same or different and are selected from O, NR.sup.z and
S, or (ii) an oxo (.dbd.O), thio (.dbd.S) or imino (.dbd.NR) group
wherein R.sup.z is R.sup.x; each occurrence of q is independently
0, 1 or 2; and each occurrence of r is independently 0, 1 or 2.
2. A compound of claim 1, wherein A and B are independently
selected from ##STR00138## wherein R.sup.3 is hydrogen, halogen or
substituted or unsubstituted C.sub.1-3 alkyl.
3. A compound of claim 1, wherein A and B are independently
selected from ##STR00139##
4. A compound of claim 1, wherein A and B are independently
selected from ##STR00140##
5. A compound of claim 1, wherein A and B are independently
selected from ##STR00141##
6. A compound of claim 1, wherein A and B are independently
selected from ##STR00142##
7. A compound of claim 1, wherein A is ##STR00143## and B is
##STR00144##
8. A compound of claim 1, wherein A is ##STR00145## and B is
##STR00146##
9. A compound of claim 1, wherein P and Q are each independently
selected from --NR.sup.xC(.dbd.O)--(CR.sup.xR.sup.y).sub.r--,
--(CR.sup.xR.sup.y).sub.r--C(.dbd.O)--NR.sup.x--,
--C(.dbd.O)NR.sup.x--(CR.sup.xR.sup.y).sub.r--,
--(CR.sup.xR.sup.y).sub.r--NR.sup.x--C(.dbd.O)--,
--NH--C(.dbd.O)--C(R.sup.xR.sup.y)--,
--(CR.sup.xR.sup.y)--C(.dbd.O)--NH--, --NR.sup.xC(.dbd.O)--,
--NR.sup.xC(.dbd.S)--, --NR.sup.xS(.dbd.O).sub.q--,
--C(.dbd.O)NR.sup.x--, --C(.dbd.S)NR.sup.x--, or --NR.sup.x--.
10. A compound of claim 1, wherein P and Q are each independently
selected from --NR.sup.xC(.dbd.O)--(CR.sup.xR.sup.y)--,
--(CR.sup.xR.sup.y)--C(.dbd.O)--NR.sup.xC(.dbd.O)-- or
--NR.sup.x--, wherein R.sup.x and R are independently selected from
hydrogen, substituted or unsubstituted C.sub.1-3 alkyl, halogen,
hydroxy and substituted or unsubstituted C.sub.1-3 alkoxy.
11. A compound of claim 1, wherein P and Q are each independently
selected from --NH--C(.dbd.O)--(CR.sup.xR.sup.y)--,
--(CR.sup.xR.sup.y)--C(.dbd.O)--NH--, --NH--C(.dbd.O)-- or --NH--,
wherein R.sup.x and R.sup.y are hydrogen.
12. A compound of claim 1, wherein (i) each of P and Q are
independently --NH--C(.dbd.O)--(CH.sub.2)--,
--(CH.sub.2)--C(.dbd.O)--NH--, --NH--C(.dbd.O)-- or --NH--; (ii) P
is --(CH.sub.2)--C(.dbd.O)--NH-- and Q is
--NH--C(.dbd.O)--CH.sub.2--, --NH--C(.dbd.O)-- or --NH--; (iii) P
is --(CH.sub.2)--C(.dbd.O)--NH--, --NH--C(.dbd.O)-- or --NH-- and Q
is --NH--C(.dbd.O)--CH.sub.2--; or (iv) P is
--(CH.sub.2)--C(.dbd.O)--NH-- and Q is
--NH--C(.dbd.O)--CH.sub.2--.
13. A compound of claim 1, wherein both P and Q are independently
selected from --NH--C(.dbd.O)--CH.sub.2-- or
--CH.sub.2--C(.dbd.O)--NH--.
14. A compound of formula (II) or (III) ##STR00147## or a tautomer
thereof, prodrug thereof, N-oxide thereof, stereoisomer thereof,
pharmaceutically acceptable ester thereof or pharmaceutically
acceptable salt thereof, wherein L is -L.sub.1-L.sub.2-L.sub.3-;
L.sub.1 is absent or independently selected from substituted or
unsubstituted C.sub.1-6 alkyl, substituted or unsubstituted
C.sub.2-6 alkenyl, substituted or unsubstituted C.sub.2-6 alkynyl,
O, S, --S(.dbd.O).sub.q--, --C(.dbd.O)-- and --NR.sup.x--; L.sub.2
is substituted or unsubstituted 3 to 14 membered heterocyclyl;
L.sub.3 is absent or independently selected from substituted or
unsubstituted C.sub.1-6 alkyl, substituted or unsubstituted
C.sub.2-6 alkenyl, substituted or unsubstituted C.sub.2-6 alkynyl,
O, S, --S(.dbd.O).sub.q--, --C(.dbd.O)-- and --NR.sup.x--; R.sup.1
and R.sup.2 are independently selected from hydrogen, hydroxy,
halogen, substituted or unsubstituted alkyl, substituted or
unsubstituted alkoxy, substituted or unsubstituted alkenyl,
substituted or unsubstituted alkynyl, substituted or unsubstituted
cycloalkyl, substituted or unsubstituted cycloalkenyl, substituted
or unsubstituted cycloalkylalkyl, substituted or unsubstituted
cycloalkenylalkyl, substituted or unsubstituted heterocyclyl,
substituted or unsubstituted heterocyclylalkyl, substituted or
unsubstituted aryl, substituted or unsubstituted arylalkyl,
substituted or unsubstituted heteroaryl, substituted or
unsubstituted heteroarylalkyl, --C(.dbd.O)OR.sup.z,
--C(.dbd.O)R.sup.z, --C(.dbd.S)R.sup.z, --C(.dbd.O)NR.sup.zR.sup.z,
--C(.dbd.O)ONR.sup.zR.sup.z, --NR.sup.zR.sup.z,
--NR.sup.zC(.dbd.O)NR.sup.zR.sup.z, --NR.sup.zS(.dbd.O)R.sup.z,
--NR.sup.zS(.dbd.O).sub.2R.sup.z, .dbd.N--NR.sup.zR.sup.z,
--NR.sup.zC(.dbd.O)OR.sup.z, --NR.sup.zC(.dbd.O)R.sup.z,
--NR.sup.xC(.dbd.S)R.sup.y --NR.sup.zC(.dbd.S)NR.sup.zR.sup.z,
--SONR.sup.zR.sup.z, --SO.sub.2NR.sup.zR.sup.z, --OR.sup.z,
--OR.sup.zC(.dbd.O)NR.sup.zR.sup.z, --OR.sup.zC(.dbd.O)OR.sup.z,
--OC(.dbd.O)R.sup.z, --OC(.dbd.O)NR.sup.zR.sup.z,
--R.sup.zNR.sup.zC(.dbd.O)R.sup.z, --R.sup.zOR.sup.z,
--R.sup.zC(.dbd.O)OR.sup.z, --R.sup.zC(.dbd.O)NR.sup.zR.sup.z,
--R.sup.zC(.dbd.O)R.sup.z, --R.sup.zOC(.dbd.O)R.sup.z, --SR.sup.z,
--SOR.sup.z, --SO.sub.2R.sup.z, --CR.sup.xR.sup.yC(.dbd.O)R.sup.z
or --CR.sup.xR.sup.yC(.dbd.S)R.sup.z; each occurrence of R.sup.x,
R.sup.y and R.sup.z is independently selected from hydrogen,
substituted or unsubstituted alkyl, substituted or unsubstituted
alkoxy, substituted or unsubstituted alkenyl, substituted or
unsubstituted alkynyl, substituted or unsubstituted aryl,
substituted or unsubstituted arylalkyl, substituted or
unsubstituted heteroaryl, substituted or unsubstituted
heteroarylalkyl, substituted or unsubstituted cycloalkyl,
substituted or unsubstituted cycloalkylalkyl, substituted or
unsubstituted cycloalkenyl, substituted or unsubstituted
heterocyclic ring, substituted heterocyclylalkyl ring, or
substituted or unsubstituted amino, or any two of R.sup.x and R
when bound to a common atom may be joined to form (i) a substituted
or unsubstituted saturated or unsaturated 3-14 membered ring, which
may optionally include one or more heteroatoms which may be the
same or different and are selected from O, NR.sup.z and S, or (ii)
an oxo (.dbd.O), thio (.dbd.S) or imino (.dbd.NR.sup.z) group;
R.sup.3 is hydrogen, halogen, substituted or unsubstituted
C.sub.1-3alkyl, nitro, amino, substituted or unsubstituted
C.sub.1-6 alkoxy, or substituted or unsubstituted C.sub.1-6
alkylamino; and each occurrence of q is independently 0, 1 or
2.
15. A compound of claim 1, wherein L.sub.1 is absent, substituted
or unsubstituted C.sub.1-6 alkyl or NR.sup.x, wherein R.sup.x is
hydrogen or C.sub.1-3 alkyl; L.sub.2 is substituted or
unsubstituted 3 to 10 membered heterocyclyl; and L.sub.3 is absent,
substituted or unsubstituted C.sub.1-6 alkyl or NR.sup.x, wherein
R.sup.x is hydrogen or C.sub.1-3 alkyl.
16. A compound of claim 1, wherein L.sub.1 is absent or substituted
or unsubstituted C.sub.1-6 alkyl; L.sub.2 is substituted or
unsubstituted 3 to 10 membered heterocyclyl; and L.sub.3 is absent
or substituted or unsubstituted C.sub.1-6 alkyl.
17. A compound of claim 1, wherein L.sub.2 is substituted or
unsubstituted 3 to 10 membered heterocyclyl, and (i) L.sub.1 and
L.sub.3 are absent; (ii) L.sub.1 and L.sub.3 are --CH.sub.2--;
(iii) L.sub.1 is --CH.sub.2-- and L.sub.3 is absent; or (iv)
L.sub.1 is absent and L.sub.3 is --CH.sub.2--.
18. A compound of claim 1, wherein L.sub.2 is selected from
##STR00148## wherein D and E are independently selected from CR''
or N, wherein R'' is hydrogen, hydroxyl, halogen or substituted or
unsubstituted C.sub.1-3 alkyl; each occurrence of R.sup.a, R.sup.b,
R.sup.c, R.sup.d, R.sup.e, R.sup.f, R.sup.g and R.sup.h is
independently selected from hydrogen, nitro, hydroxy, cyano,
halogen, substituted or unsubstituted C.sub.1-6 alkyl, substituted
or unsubstituted C.sub.2-6 alkenyl, substituted or unsubstituted
C.sub.2-6 alkynyl, substituted or unsubstituted C.sub.3-6
cycloalkyl, substituted or unsubstituted C.sub.3-6 cycloalkylalkyl,
and substituted or unsubstituted C.sub.3-6 cycloalkenyl; or any two
of R.sup.a, R.sup.b, R.sup.c, R.sup.d, R.sup.e, R.sup.f, R.sup.g
and R.sup.h may be joined to form (i) a substituted or
unsubstituted, saturated or unsaturated 3-14 membered ring, which
may optionally include one or more heteroatoms which may be the
same or different and are selected from O, NR' (where R' is H or
C.sub.1-3 alkyl) and S, or (ii) an oxo (.dbd.O), thio (.dbd.S) or
imino (.dbd.NR') group; and each of s, t, u and v is independently
0, 1 or 2 with the proviso that the sum of s, t, u and v is not
0.
19. A compound of claim 18, wherein each occurrence of R.sup.a,
R.sup.b, R.sup.c, R.sup.d, R.sup.e, R.sup.f, R.sup.g and R.sup.h is
independently selected from hydrogen, hydroxy, substituted or
unsubstituted C.sub.1-3 alkyl or any two of R.sup.a, R.sup.b,
R.sup.c, R.sup.d, R.sup.e, R.sup.f, R.sup.g and R.sup.h when bound
to a common atom form an oxo (.dbd.O) or substituted or
unsubstituted cycloalkyl.
20. A compound of claim 19, wherein each of R.sup.a, R.sup.b,
R.sup.c, R.sup.d, R.sup.e, R.sup.f, R.sup.g and R.sup.h is hydrogen
and the sum of s, t, u and v is 1, 2, 3 or 4.
21. A compound of claim 20, wherein (i) each of R.sup.a, R.sup.b,
R.sup.c, R.sup.d, R.sup.e, R.sup.f, R.sup.g and R.sup.h is
hydrogen, s is 0, t is 1 and u+v=3; (ii) each of R.sup.a, R.sup.b,
R.sup.c, R.sup.d, R.sup.e, R.sup.f, R.sup.g and R.sup.h is hydrogen
and s, t, u and v are each 1; (iii) each of R.sup.a, R.sup.b,
R.sup.c, R.sup.d, R.sup.e, R.sup.f, R.sup.g and R.sup.h is
hydrogen, s is 0 and t, u and v are each 1; or (iv) each of
R.sup.a, R.sup.b, R.sup.c, R.sup.d, R.sup.e, R.sup.f, R.sup.g and
R.sup.h is hydrogen and the sum of s, t, u and v is 2; or (v) each
of R.sup.a, R.sup.b, R.sup.c, R.sup.d, R.sup.e, R.sup.f, R.sup.g
and R.sup.h is hydrogen and the sum of s, t, u and v is 1.
22. A compound of claim 18, wherein D and E are independently
selected from CH and N.
23. A compound of claim 18, wherein (i) D is CH and E is N or (ii)
D is N and E is CH.
24. A compound of claim 1, wherein each of L.sub.1 and L.sub.3 is
absent or selected from substituted or unsubstituted C.sub.1-3
alkyl.
25. A compound of claim 1, wherein L (L.sub.1-L.sub.2-L.sub.3) is
selected from ##STR00149## ##STR00150##
26. A compound of claim 1, wherein L.sub.2 is selected from
##STR00151##
27. A compound of claim 1, wherein L.sub.2 is selected from
##STR00152##
28. A compound of claim 1, wherein each of L.sub.1 and L.sub.3 is,
independently, absent or --CH.sub.2--.
29. A compound of claim 1, wherein R.sup.3 is hydrogen, halogen or
substituted or unsubstituted C.sub.1-3 alkyl.
30. A compound of claim 1, wherein R.sup.3 is hydrogen, fluorine or
methyl.
31. A compound of claim 1, wherein R.sup.1 and R.sup.2 are each
independently selected from hydrogen, halogen, substituted or
unsubstituted alkyl, --NR.sup.zR.sup.z, substituted or
unsubstituted heterocyclyl, substituted or unsubstituted
heterocyclylalkyl, substituted or unsubstituted aryl, substituted
or unsubstituted arylalkyl, substituted or unsubstituted
heteroaryl, and substituted or unsubstituted heteroarylalkyl.
32. A compound of claim 1, wherein R.sup.1 and R.sup.2 are each
independently selected from --NR.sup.zR.sup.z, substituted or
unsubstituted aryl and substituted or unsubstituted heteroaryl.
33. A compound of claim 1, wherein R.sup.1 and R.sup.2 are each
independently selected from substituted or unsubstituted aryl and
substituted or unsubstituted heteroaryl.
34. A compound of claim 1, wherein (i) R.sup.1 is substituted or
unsubstituted aryl and R.sup.2 is substituted or unsubstituted
heteroaryl; (ii) R.sup.1 is substituted or unsubstituted heteroaryl
and R.sup.2 is substituted or unsubstituted aryl; (iii) both
R.sup.1 and R.sup.2 are substituted or unsubstituted aryl or (iv)
both R.sup.1 and R.sup.2 are substituted or unsubstituted
heteroaryl.
35. A compound of claim 1, wherein R.sup.1 and R.sup.2 are each
independently selected from ##STR00153## ##STR00154##
36. A compound of claim 1, wherein R.sup.x and R.sup.y are
hydrogen, hydroxyl or --CH.sub.2OH.
37. A compound selected from:
2-(Pyridin-2-yl)-N-(5-(1-(6-(2-(3-(trifluoromethoxy)phenyl)acetamido)pyri-
din-3-yl) piperidin-4-yl)-1,3,4-thiadiazol-2-yl)acetamide;
(RS)-2-(Pyridin-2-yl)-N-(5-(1-(6-(2-(3-(trifluoromethoxy)phenyl)acetamido-
)pyridin-3-yl) piperidin-3-yl)-1,3,4-thiadiazol-2-yl)acetamide;
(R)-2-(Pyridin-2-yl)-N-(5-(1-(6-(2-(3-(trifluoromethoxy)phenyl)acetamido)
pyridin-3-yl) piperidin-3-yl)-1,3,4-thiadiazol-2-yl)acetamide;
(S)-2-(Pyridin-2-yl)-N-(5-(1-(6-(2-(3-(trifluoromethoxy)phenyl)acetamido)
pyridin-3-yl) piperidin-3-yl)-1,3,4-thiadiazol-2-yl)acetamide;
(RS)-2-(Pyridin-3-yl)-N-(5-(1-(6-(2-(3-(trifluoromethoxy)phenyl)acetamido-
)pyridin-3-yl) piperidin-3-yl)-1,3,4-thiadiazol-2-yl)acetamide;
(R)-2-(Pyridin-3-yl)-N-(5-(1-(6-(2-(3-(trifluoromethoxy)phenyl)acetamido)
pyridin-3-yl) piperidin-3-yl)-1,3,4-thiadiazol-2-yl)acetamide;
(S)-2-(Pyridin-3-yl)-N-(5-(1-(6-(2-(3-(trifluoromethoxy)phenyl)acetamido)
pyridin-3-yl) piperidin-3-yl)-1,3,4-thiadiazol-2-yl)acetamide;
2-(Pyridin-3-yl)-N-(5-(1-(6-(2-(3-(trifluoromethoxy)phenyl)acetamido)pyri-
din-3-yl) piperidin-4-yl)-1,3,4-thiadiazol-2-yl)acetamide;
2-(3-Cyanophenyl)-N-(5-(1-(6-(2-(3-(trifluoromethoxy)phenyl)acetamido)pyr-
idin-3-yl)piperidin-4-yl)-1,3,4-thiadiazol-2-yl)acetamide;
2-(Pyridin-2-yl)-N-(5-(4-(5-(2-(3-(trifluoromethoxy)phenyl)acetamido)-1,3-
,4-thiadiazol-2-yl)piperidin-1-yl)pyridin-2-yl)acetamide;
2-(Pyridin-2-yl)-N-(5-(3-(5-(2-(3-(trifluoromethoxy)phenyl)acetamido)-1,3-
,4-thiadiazol-2-yl)piperidin-1-yl)pyridin-2-yl)acetamide;
2-(pyridin-2-yl)-N-(5-(1-(6-(2-(3-(trifluoromethoxy)phenyl)acetamido)pyri-
dazin-3-yl) piperidin-4-yl)-1,3,4-thiadiazol-2-yl)acetamide;
2-(Pyridin-2-yl)-N-(5-(1-(6-(2-(3-(trifluoromethoxy)phenyl)acetamido)pyri-
dazin-3-yl)piperidin-3-yl)-1,3,4-thiadiazol-2-yl)acetamide;
2-(Pyridin-3-yl)-N-(5-(1-(6-(2-(3-(trifluoromethoxy)phenyl)acetamido)pyri-
dazin-3-yl)piperidin-4-yl)-1,3,4-thiadiazol-2-yl)acetamide;
2-(3-(Methylsulfonamido)phenyl)-N-(5-(1-(6-(2-(3-(trifluoromethoxy)phenyl-
)
acetamido)pyridazin-3-yl)piperidin-4-yl)-1,3,4-thiadiazol-2-yl)acetamide-
;
2-(2-Chlorophenyl)-N-(6-(4-(5-(2-(pyridin-2-yl)acetamido)-1,3,4-thiadiaz-
ol-2-yl) piperidin-1-yl)pyridazin-3-yl)acetamide;
2-(2-Chlorophenyl)-N-(5-(1-(6-(2-(3-(trifluoromethoxy)phenyl)acetamido)py-
ridazin-3-yl)piperidin-3-yl)-1,3,4-thiadiazol-2-yl)acetamide;
2-(2-Fluorophenyl)-N-(6-(4-(5-(2-(pyridin-2-yl)acetamido)-1,3,4-thiadiazo-
l-2-yl) piperidin-1-yl)pyridazin-3-yl)acetamide;
2-(Pyrazin-2-yl)-N-(5-(1-(6-(2-(3-(trifluoromethoxy)phenyl)acetamido)pyri-
dazin-3-yl)piperidin-4-yl)-1,3,4-thiadiazol-2-yl)acetamide;
2-(Pyridin-2-yl)-N-(5-(1-(6-(2-(3-(trifluoromethoxy)phenyl)acetamido)pyri-
dazin-3-yl)piperidin-4-yl)-1,3,4-thiadiazol-2-yl)acetamide
dihydrochloride;
2-(Pyridin-2-yl)-N-(6-(4-(5-(2-(3-(trifluoromethoxy)phenyl)acetamido)-1,3-
,4-thiadiazol-2-yl)piperidin-1-yl)pyridazin-3-yl)acetamide;
2-(Pyridin-3-yl)-N-(6-(4-(5-(2-(3-(trifluoromethoxy)phenyl)acetamido)-1,3-
,4-thiadiazol-2-yl)piperidin-1-yl)pyridazin-3-yl)acetamide;
2-(Pyridin-3-yl)-N-(6-(4-(5-(2-(2,3,6-trifluorophenyl)acetamido)-1,3,4-th-
iadiazol-2-yl)piperidin-1-yl)pyridazin-3-yl)acetamide;
2-(Pyridin-2-yl)-N-(6-(4-(5-(2-(2,3,6-trifluorophenyl)acetamido)-1,3,4-th-
iadiazol-2-yl)piperidin-1-yl)pyridazin-3-yl)acetamide;
2-(2,3-Difluorophenyl)-N-(5-(1-(6-(2-(pyridin-2-yl)acetamido)pyridazin-3--
yl)piperidin-4-yl)-1,3,4-thiadiazol-2-yl)acetamide;
2-(3,4-Difluorophenyl)-N-(5-(1-(6-(2-(pyridin-2-yl)acetamido)pyridazin-3--
yl)piperidin-4-yl)-1,3,4-thiadiazol-2-yl)acetamide;
2-(2-Fluorophenyl)-N-(5-(1-(6-(2-(pyridin-2-yl)acetamido)pyridazin-3-yl)p-
iperidin-4-yl)-1,3,4-thiadiazol-2-yl)acetamide;
2-(3-Fluorophenyl)-N-(5-(1-(6-(2-(pyridin-2-yl)acetamido)p;yridazin-3-yl)-
piperidin-4-yl)-1,3,4-thiadiazol-2-yl)acetamide;
2-(4-Fluorophenyl)-N-(5-(1-(6-(2-(pyridin-2-yl)acetamido)pyridazin-3-yl)p-
iperidin-4-yl)-1,3,4-thiadiazol-2-yl)acetamide;
2-(3-Chlorophenyl)-N-(5-(1-(6-(2-(pyridin-2-yl)acetamido)pyridazin-3-yl)p-
iperidin-4-yl)-1,3,4-thiadiazol-2-yl)acetamide;
3-Hydroxy-2-phenyl-N-(5-(1-(6-(2-(3-(trifluoromethoxy)phenyl)acetamido)
pyridazin-3-yl)piperidin-4-yl)-1,3,4-thiadiazol-2-yl)propanamide;
(R)-2-hydroxy-2-phenyl-N-(5-(1-(6-(2-(3-(trifluoromethoxy)phenyl)acetamid-
o) pyridazin-3-yl)piperidin-4-yl)-1,3,4-thiadiazol-2-yl)acetamide;
2-(3-(3-Fluoroazetidin-1-yl)phenyl)-N-(5-(1-(6-(2-(3-(trifluoromethoxy)ph-
enyl)
acetamido)pyridazin-3-yl)piperidin-4-yl)-1,3,4-thiadiazol-2-yl)aceta-
mide;
2-(Pyridin-2-yl)-N-(5-((1-(6-(2-(3-(trifluoromethoxy)phenyl)acetamid-
o)pyridazin-3-yl)
piperidin-4-yl)methyl)-1,3,4-thiadiazol-2-yl)acetamide; and
pharmaceutically acceptable salts thereof.
38. A pharmaceutical composition comprising a compound of claim 1
and a pharmaceutically acceptable carrier.
39-48. (canceled)
Description
[0001] The present application claims the benefit of Indian Patent
Application Nos. 36/CHE/2014, filed Jan. 6, 2014, 39/CHE/2014,
filed Jan. 6, 2014, 2639/CHE/2014, filed May 29, 2014,
2647/CHE/2014, filed May 29, 2014, 2783/CHE/2014, filed Jun. 6,
2014, 3525/CHE/2014 filed Jul. 18, 2014, 3612/CHE/2014, filed Jul.
24, 2014, 3613/CHE/2014, filed Jul. 24, 2014, and 5438/CHE/2014,
filed Oct. 31, 2014, each of which is hereby incorporated by
reference in its entirety.
FIELD OF THE INVENTION
[0002] The present invention provides compounds of formula (I) to
(III) as glutaminase inhibitors, methods of preparing them,
pharmaceutical compositions containing them and methods of
treatment, prevention and/or amelioration of diseases or disorders
involving glutamine.
BACKGROUND OF THE INVENTION
[0003] Glutaminase (glutaminase I, L-glutaminase, glutamine
aminohydrolase) is an amido-hydrolase enzyme that generates
glutamate from glutanine. Glutaminase has been reported to have
tissue-specific isoenzymes. Glutaminase has an important role in
glial cells. Glutamine is the most abundant free amino acid in the
human body; it is essential for the growth of normal and neoplastic
cells and for the culture of many cell types. Glutamine is an
important source of energy for neoplastic tissues, and products of
its metabolism include, among others, glutamate (Glu) and
glutathione (GSH), the two molecules that play a key role in tumor
proliferation, invasiveness and resistance to therapy. Glutamine
hydrolysis in normal and transforming mammalian tissues alike is
carried out by different isoforms of glutaminases, of which the two
major types are liver-type glutaminase (LGA) and kidney-type
glutaminase (KGA) (see Neurochem Int., 2009 July-August;
55(1-3):71-5. doi: 10.1016/j.neuint.2009.01.008. Epub 2009
February).
[0004] Cancer cells require a robust supply of reduced nitrogen to
produce nucleotides, non-essential amino acids and a high cellular
redox activity. Glutamine provides a major substrate for
respiration as well as nitrogen for the production of proteins,
hexosamines, and macromolecules. Therefore, glutamine is one of key
molecules in cancer metabolism during cell proliferation. The
notion of targeting glutamine metabolism in cancer, originally
rationalized by the number of pathways fed by this nutrient, has
been reinforced by more recent studies demonstrating that its
metabolism is regulated by oncogenes. Glutaminase (GA) is the first
enzyme that converts glutanine to glutamate, which is in turn
converted to alpha-ketoglutarate for further metabolism in the
tricarboxylic acid cycle. Different GA isoforms in mammals are
encoded by two genes, Gls and Gls2. As each enzymatic form of GA
has distinct kinetic and molecular characteristics, it has been
speculated that the differential regulation of GA isoforms may
reflect distinct functions or requirements in different tissues or
cell states. GA encoded by the Gls gene (GLS) has been demonstrated
to be regulated by oncogenes and to support tumor cell growth. GA
encoded by the Gls2 gene (GLS2) reduces cellular sensitivity to
reactive oxygen species associated apoptosis possibly through
glutathione-dependent antioxidant defense, and therefore to behave
more like a tumor suppressor. Thus, modulation of GA function may
be a new therapeutic target for cancer treatment (see Mates et al.,
Curr. Mol. Med., 2013 May; 13(4), 514-534).
[0005] One hallmark of cancer cells is their adaptation to rely
upon an altered metabolic scheme that includes changes in the
glycolytic pathway, known as the Warburg effect, and elevated
glutamine metabolism. Glutaminase, a mitochondrial enzyme, plays a
key role in the metabolism of glutamine in cancer cells, and its
inhibition could significantly impact malignant transformation (see
Katt et al., Mol. Cancer Ther., 11(6); 1269-78, 2012). Feeding off
the breakdown of glutamine, cancer cells are able to grow and
divide into a tumour. Glutaminase therefore makes a promising
therapeutic target for the prevention of tumour progression.
Inhibition of this enzyme could effectively starve the cancer cells
of their energy source. See Medina et al., J. Nutr., Sep. 1, 2001,
Vol. 131, No. 9 2539-25425.
[0006] Glutaminase plays a crucial role in the mechanisms of
cancer, such as cell survival, proliferation and growth. There are
two reported glutaminase inhibitors namely
6-diazo-5-oxo-L-norleucine (DON) which was isolated originally from
Streptomyces in a sample of Peruvian soil and was characterized in
1956 by Henry W Dion (see Dion et al., Antibiotics and
Chemotherapy, 1954, 78, 3075-3077) and suggested as a cancer
therapy and bis-2-(5-phenylacetamido-1,3,4-thiadiazol-2-yl)ethyl
sulfide (BPTES) disclosed by Elan Pharmaceuticals. There is an
ongoing research effort reported by other groups working at Cornell
University and Calithera Biosciencesis to discover and identify
small molecule inhibitors of glutaminase. DON has also been
reported to be evaluated in combination with PEG-PGA by New Medical
Enzymes AG. In addition to BPTES and DON, other reported
glutaminase inhibitors are as summarised in the table below.
TABLE-US-00001 Agent Company Clinical status BPTES -- Discovery
CB-839 Calithera Biosciences Phase-1 Compound 968 Cornell
University Preclinic al GlutaDON New Medical Phase-2 (PEG-PGA +
DON) Enzymes AG GlutaChemo New Medical Preclinical (PEG-PGA + an
Enzymes AG ideal candidate)
[0007] Reviews and studies regarding Glutamine and Glutaminase in
cancer and other discesase have been given by Medina et al., J.
Nutr., Sep. 1, 2001, Vol. 131, No. 9, 2539S-2542S; Ajit G. Thmas et
al., Biochemical and Biophysical Research Communications, 443,
2014, 32-36; Monica Szeliga et al., Neurochemistrt Intermationa,
55, 2009, 71-77; and Curthoys et al., Annu. Rev. Nutr., 1995, 15,
133-159. All of these literature disclosures are incorporated
herein by reference in their entirety for all purposes.
[0008] Patent literature related to glutaminase inhibitors includes
International Publication Nos. WO 99/09825, WO 00/59533, WO
03/022261, WO 04/108153, WO 07/128588, WO 10/033871, WO 10/111504,
WO 11/076967, WO 11/143160, WO 12/006506, WO 12/034123, WO
13/044596, WO 13/078123, WO 14/078645, WO 14/089048, WO14/043633,
WO14/079011, WO14/079136, WO14/079150, and WO14/081925, U.S.
Publication Nos. 2002/0115698, 2006/0276438, 2013/0157998,
2014/0050699, 2014/0194421, 2014/0369961, 2015/0004134,
20140142081, and 20140142146, U.S. Pat. Nos. 5,552,427, 6,451,828,
8,465,736, 8,604,016, and 8,865,718 and European Publication No.
656210, each of which is incorporated herein by reference in its
entirety for all purposes.
[0009] There still remains an unmet need for novel glutaminase
inhibitors for the treatment of diseases and disorders associated
with cell proliferation such as cancer and other immunological and
neurological disorders.
SUMMARY OF THE INVENTION
[0010] The present invention relates to compounds of formula (I) to
(III), methods for their preparation, pharmaceutical compositions
containing them, and methods of treatment using them. In
particular, the compounds of formula (I) to (III) and their
pharmaceutically acceptable salts are useful in the treatment,
prevention and/or amelioration of diseases or disorders involving
glutamine.
[0011] In one aspect, the present invention relates to a compound
of formula (I):
##STR00001##
or a tautomer thereof, prodrug thereof, N-oxide thereof,
stereoisomer thereof, pharmaceutically acceptable ester thereof or
pharmaceutically acceptable salt thereof, wherein
[0012] L is -L.sub.1-L.sub.2-L.sub.3-; wherein
[0013] L.sub.1 is absent or independently selected from substituted
or unsubstituted C.sub.1-6 alkyl, substituted or unsubstituted
C.sub.2-6 alkenyl, substituted or unsubstituted C.sub.2-6 alkynyl,
O, S, --S(.dbd.O).sub.q--, --C(.dbd.O)-- and --NR.sup.x--;
[0014] L.sub.2 is substituted or unsubstituted 3 to 14 membered
heterocyclyl;
[0015] L.sub.3 is absent or independently selected from substituted
or unsubstituted C.sub.1-6 alkyl, substituted or unsubstituted
C.sub.2-6 alkenyl, substituted or unsubstituted C.sub.2-6 alkynyl,
O, S, --S(.dbd.O).sub.q--, --C(.dbd.O)-- and --NR.sub.x--;
[0016] A and B are independently selected from
##STR00002##
each of A and B are optionally substituted with one or more
R.sup.3;
[0017] each occurrence of R.sup.3 is, independently, hydrogen,
halogen, substituted or unsubstituted C.sub.1-3 alkyl, nitro,
amino, substituted or unsubstituted C.sub.1-6 alkoxy, or
substituted or unsubstituted C.sub.1-6 alkylamino;
[0018] P and Q are independently selected from substituted or
unsubstituted alkyl, substituted or unsubstituted alkenyl,
substituted or unsubstituted alkynyl, substituted or unsubstituted
alkoxy, --NR.sup.x--C(.dbd.O)--(CR.sup.xR.sup.y).sub.r--,
--NH--C(.dbd.O)--C(R.sup.xR.sup.y)--,
--(CR.sup.xR.sup.y).sub.r--C(.dbd.O)--NR.sup.x--,
--(CR.sup.xR.sup.y)--C(.dbd.O)--NH--,
--C(.dbd.O)NR.sup.x--(CR.sup.xR.sup.y).sub.r--,
--C(.dbd.O)NH--C(R.sup.xR.sup.y)--,
--(CR.sup.xR.sup.y).sub.r--NR.sup.x--C(.dbd.O)--,
--(CR.sup.xR.sup.y)--NH--C(.dbd.O)--, --NR.sup.x--,
--NR.sup.xC(.dbd.O)--, --NR.sup.xC(.dbd.S)--,
--NR.sup.xS(.dbd.O).sub.q--, --C(.dbd.O)NR.sup.x--,
--C(.dbd.S)NR.sup.x--, --S(.dbd.O).sub.qNR.sup.x--,
--NR.sup.xC(.dbd.O)NR.sup.x--, --NR.sup.xC(.dbd.S)NR.sup.x--,
--C(.dbd.O)--, --C(.dbd.S)--, --C(.dbd.O)ONR.sup.x--,
.dbd.N--N(R.sup.x)--, --N(R.sup.x)--N.dbd. or
--NR.sup.xC(.dbd.O)O--;
[0019] R.sup.1 and R.sup.2 are independently selected from
hydrogen, hydroxy, halogen, substituted or unsubstituted alkyl,
substituted or unsubstituted alkoxy, substituted or unsubstituted
alkenyl, substituted or unsubstituted alkynyl, substituted or
unsubstituted cycloalkyl, substituted or unsubstituted
cycloalkenyl, substituted or unsubstituted cycloalkylalkyl,
substituted or unsubstituted cycloalkenylalkyl, substituted or
unsubstituted heterocyclyl, substituted or unsubstituted
heterocyclylalkyl, substituted or unsubstituted aryl, substituted
or unsubstituted arylalkyl, substituted or unsubstituted
heteroaryl, substituted or unsubstituted heteroarylalkyl,
--C(.dbd.O)OR.sup.z, --C(.dbd.O)R.sup.z, --C(.dbd.S)R.sup.z,
--C(.dbd.O)NR.sup.zR.sup.z, --C(.dbd.O)ONR.sup.zR.sup.z,
--NR.sup.zR.sup.z, --NR.sup.zC(.dbd.O)NR.sup.zR.sup.z,
--NR.sup.zS(.dbd.O)R.sup.z, --NR.sup.zS(.dbd.O).sub.2R.sup.z,
.dbd.N--N--R.sup.zR.sup.z, --NR.sup.zC(.dbd.O)OR.sup.z,
--NR.sup.zC(.dbd.O)R.sup.z, --NR.sup.xC(.dbd.S)R.sup.y,
--NR.sup.zC(.dbd.S)NR.sup.zR.sup.z, --SONR.sup.zR.sup.z,
--SO.sub.2NR.sup.zR.sup.z, --OR.sup.z,
--OR.sup.zC(.dbd.O)NR.sup.zR.sup.z, --OR.sup.zC(.dbd.O)OR.sup.z,
--OC(.dbd.O)R.sup.z, --OC(.dbd.O)NR.sup.zR.sup.z,
--R.sup.zNR.sup.zC(.dbd.O)R.sup.z, --R.sup.zOR.sup.z,
--R.sup.zC(.dbd.O)OR.sup.z, --R.sup.zC(.dbd.O)NR.sup.zR.sup.z,
--R.sup.zC(.dbd.O)R.sup.z, --ROC(.dbd.O)R.sup.z, --SR.sup.z,
--SOR.sup.z, --SO.sub.2R.sup.z, --CR.sup.xR.sup.yC(.dbd.O)R.sup.z
or --CR.sup.xR.sup.yC(.dbd.S)R.sup.z;
[0020] each occurrence of R.sup.x, R.sup.y and R.sup.z is
independently selected from hydrogen, substituted or unsubstituted
alkyl, substituted or unsubstituted alkoxy, substituted or
unsubstituted alkenyl, substituted or unsubstituted alkynyl,
substituted or unsubstituted aryl, substituted or unsubstituted
arylalkyl, substituted or unsubstituted heteroaryl, substituted or
unsubstituted heteroarylalkyl, substituted or unsubstituted
cycloalkyl, substituted or unsubstituted cycloalkylalkyl,
substituted or unsubstituted cycloalkenyl, substituted or
unsubstituted heterocyclic ring, substituted heterocyclylalkyl
ring, or substituted or unsubstituted amino, or any two of R.sup.x
and R.sup.y when bound to a common atom may be joined to form (i) a
substituted or unsubstituted saturated or unsaturated 3-14 membered
ring, which may optionally include one or more heteroatoms which
may be the same or different and are selected from O, NR.sup.z and
S, or (ii) an oxo (.dbd.O), thio (.dbd.S) or imino (.dbd.NR.sup.z)
group;
[0021] each occurrence of q is independently 0, 1 or 2; and
[0022] each occurrence of r is independently 0, 1 or 2.
[0023] For avoidance of doubt and unless indicated otherwise,
formulas are to be read in the direction they are shown. For
example, (a) if P is --CH.sub.2--C(.dbd.O)--NH-- in formula (I)
(i.e., R.sup.1--P-A-L-B-Q-R.sup.2) than the compound would have the
formula R.sup.1--CH.sub.2--C(O)--NH-A-L-B-Q-R.sup.2 or (b) if P is
--CH.sub.2--C(.dbd.O)--NH-- and Q is --NH--C(.dbd.O)--CH.sub.2-- in
formula (I) (i.e., R.sup.1--P-A-L-B-Q-R.sup.2) than the compound
would have the formula
R.sup.1--CH.sub.2--C(O)--NH-A-L-B--NH--C(.dbd.O)--CH.sub.2--R.sup.2.
[0024] Further preferred is a compound having the formula (I),
wherein A and B are independently selected from
##STR00003##
wherein R.sup.3 is hydrogen, halogen or substituted or
unsubstituted C.sub.1-3 alkyl (e.g., methyl).
[0025] Further preferred is a compound having the formula (I),
wherein A and B are independently selected from
##STR00004##
[0026] Further preferred is a compound having the formula (I),
wherein A and B are independently selected from
##STR00005##
[0027] Further preferred is a compound having the formula (I),
wherein A and B are independently selected from
##STR00006##
[0028] Further preferred is a compound having the formula (I),
wherein A and B are independently selected from
##STR00007##
[0029] Further preferred is a compound having the formula (I),
wherein [0030] A is
##STR00008##
[0030] and B is
##STR00009##
[0032] Further preferred is a compound having the formula (I),
wherein [0033] A is
##STR00010##
[0033] and B is
##STR00011##
[0035] Further preferred is a compound having the formula (I),
wherein each of P and Q are independently selected from
--NR.sup.xC(.dbd.O)--(CR.sup.xR.sup.y).sub.r--,
--(CR.sup.xR.sup.y).sub.r--C(.dbd.O)--NR.sup.x--,
--C(.dbd.O)NR.sup.x--(CR.sup.xR.sup.y).sub.r--,
--(CR.sup.xR.sup.y).sub.r--NR.sup.x--C(.dbd.O)--,
--NH--C(.dbd.O)--C(R.sup.xR.sup.y)--,
--(CR.sup.xR.sup.y)--C(.dbd.O)--NH--, --NR.sup.xC(.dbd.O)--,
--NR.sup.xC(.dbd.S)--, --NR.sup.xS(.dbd.O).sub.q--,
--C(.dbd.O)NR.sup.z--, --C(.dbd.S)NR.sup.z--, or --NR.sup.x--.
[0036] Further preferred is a compound having the formula (I),
wherein each of P and Q are independently selected from
--NR.sup.xC(.dbd.O)--(CR.sup.xR.sup.y)--,
--(CR.sup.xR.sup.y)--C(.dbd.O)--NR.sup.x--, --NR.sup.xC(.dbd.O)--
or --NR.sup.x--, wherein R.sup.x and R.sup.y are independently
selected from hydrogen, substituted or unsubstituted C.sub.1-3
alkyl, halogen, hydroxy and substituted or unsubstituted C.sub.1-3
alkoxy.
[0037] Further preferred is a compound having the formula (I),
wherein each of P and Q are independently
--NH--C(.dbd.O)--(CR.sup.xR.sup.y)--,
--(CR.sup.xR.sup.y)--C(.dbd.O)--NH--, --NH--C(.dbd.O)-- or --NH--,
wherein R.sup.x and R.sup.y are hydrogen.
[0038] Further preferred is a compound having the formula (I),
wherein each of P and Q are independently
--NH--C(.dbd.O)--(CH.sub.2)--, --(CH.sub.2)--C(.dbd.O)--NH--,
--NH--C(.dbd.O)-- or --NH--.
[0039] Further preferred is a compound having the formula (I),
wherein P is --(CH.sub.2)--C(.dbd.O)--NH-- and Q is
--NH--C(.dbd.O)--CH.sub.2--, --NH--C(.dbd.O)-- or --NH--.
[0040] Further preferred is a compound having the formula (I),
wherein P is --(CH.sub.2)--C(.dbd.O)--NH--, --NH--C(.dbd.O)-- or
--NH-- and Q is --NH--C(.dbd.O)--CH.sub.2--.
[0041] Further preferred is a compound having the formula (I),
wherein P is --(CH.sub.2)--C(.dbd.O)--NH-- and Q is
--NH--C(.dbd.O)--CH.sub.2--.
[0042] Yet another embodiment is a compound having the formula (II)
or (III):
##STR00012##
or a tautomer thereof, prodrug thereof, N-oxide thereof,
stereoisomer thereof, pharmaceutically acceptable ester thereof or
pharmaceutically acceptable salt thereof, wherein each of the
variables L, R.sup.1, R.sup.2, R.sup.3, R.sup.x and R.sup.y are as
defined above in relation to formula (I).
[0043] Further preferred is a compound having the formula (I), (II)
or (III), wherein [0044] L.sub.1 is absent, substituted or
unsubstituted C.sub.1-6 alkyl or --NR.sup.x--, wherein R.sup.x is
hydrogen or C.sub.1-3 alkyl; [0045] L.sub.2 is substituted or
unsubstituted 3 to 10 membered heterocyclyl; and [0046] L.sub.3 is
absent, substituted or unsubstituted C.sub.1-6 alkyl or
--NR.sup.x--, wherein R.sup.x is hydrogen or C.sub.1-3 alkyl.
[0047] Further preferred is a compound having the formula (I), (II)
or (III), wherein [0048] L.sub.1 is absent or substituted or
unsubstituted C.sub.1-6 alkyl; [0049] L.sub.2 is substituted or
unsubstituted 3 to 10 membered heterocyclyl; and [0050] L.sub.3 is
absent or substituted or unsubstituted C.sub.1-6 alkyl.
[0051] Further preferred is a compound having the formula (I), (II)
or (III), wherein [0052] L.sub.1 is substituted or unsubstituted
C.sub.1-6 alkyl; [0053] L.sub.2 is substituted or unsubstituted 3
to 10 membered heterocyclyl; and [0054] L.sub.3 is substituted or
unsubstituted C.sub.1-6 alkyl.
[0055] Further preferred is a compound having the formula (I), (II)
or (III), wherein [0056] L.sub.1 is substituted or unsubstituted
C.sub.1-6 alkyl; [0057] L.sub.2 is substituted or unsubstituted 3
to 10 membered heterocyclyl; and [0058] L.sub.3 is absent.
[0059] Further preferred is a compound having the formula (I), (II)
or (III), wherein [0060] L.sub.1 is absent. [0061] L.sub.2 is
substituted or unsubstituted 3 to 10 membered heterocyclyl; and
[0062] L.sub.3 is substituted or unsubstituted C.sub.1-6 alkyl.
[0063] Further preferred is a compound having the formula (I), (II)
or (III), wherein [0064] L.sub.1 is absent. [0065] L.sub.2 is
substituted or unsubstituted 3 to 10 membered heterocyclyl; and
[0066] L.sub.3 is absent.
[0067] Further preferred is a compound having the formula (I), (II)
or (III), wherein L.sub.2 is selected from
##STR00013##
wherein
[0068] D and E are independently selected from CR'' or N, wherein
each occurrence of R'' is independently hydrogen, hydroxyl, halogen
or substituted or unsubstituted C.sub.1-3 alkyl;
[0069] each occurrence of R.sup.a, R.sup.b, R.sup.c, R.sup.d,
R.sup.e, R.sup.f, R.sup.g and R.sup.h is independently selected
from hydrogen, nitro, hydroxy, cyano, halogen, substituted or
unsubstituted C.sub.1-6 alkyl, substituted or unsubstituted
C.sub.2-6 alkenyl, substituted or unsubstituted C.sub.2-6 alkynyl,
substituted or unsubstituted C.sub.3-6 cycloalkyl, substituted or
unsubstituted C.sub.3-6 cycloalkylalkyl, and substituted or
unsubstituted C.sub.3-6 cycloalkenyl; or any two of R.sup.a,
R.sup.b, R.sup.c, R.sup.d, R.sup.e, R.sup.f, R.sup.g and R.sup.h
(such as two groups bound to a common atom or adjacent atoms or any
two groups which when joined form a chemically stable structure)
may be joined to form (i) a substituted or unsubstituted, saturated
or unsaturated 3-14 membered ring, which may optionally include one
or more heteroatoms which may be the same or different and are
selected from O, NR' (where R' is H or C.sub.1-3 alkyl) and S, or
(ii) an oxo (.dbd.O), thio (.dbd.S) or imino (.dbd.NR') group; and
each of s, t, u and v is 0, 1 or 2 with the proviso that the sum of
s, t, u and v is not O (i.e., s+t+u+v.gtoreq.1).
[0070] Further preferred is a compound having the formula (I), (II)
or (III), wherein each occurrence of R.sup.a, R.sup.b, R.sup.c,
R.sup.d, R.sup.e, R.sup.f, R.sup.g and R.sup.h is independently
selected from hydrogen, hydroxy, and substituted or unsubstituted
C.sub.1-3 alkyl, or any two of R.sup.a, R.sup.b, R.sup.c, R.sup.d,
R.sup.e, R.sup.f, R.sup.g and R.sup.h when bound to a common atom
may form oxo (.dbd.O) or any two of R.sup.a, R.sup.b, R.sup.c,
R.sup.d, R.sup.e, R.sup.f, R.sup.g and R.sup.h form a substituted
or unsubstituted cycloalkyl group.
[0071] Further preferred is a compound having the formula (I), (II)
or (III), wherein each of R.sup.a, R.sup.b, R.sup.c, R.sup.d,
R.sup.e, R.sup.f, R.sup.g and R.sup.h is hydrogen and the sum of s,
t, u and v is 1-4, such as 1, 2, 3 or 4.
[0072] Further preferred is a compound having the formula (I), (II)
or (111), wherein R.sup.a, R.sup.b, R.sup.c, R.sup.d, R.sup.e,
R.sup.f, R.sup.g and R.sup.h is hydrogen, s is 0, t is 1 and the
sum of u and v is 3.
[0073] Further preferred is a compound having the formula (I), (II)
or (III), wherein [0074] (i) R.sup.a, R.sup.b, R.sup.c, R.sup.d,
R.sup.e, R.sup.f, R.sup.g and R.sup.h is hydrogen, and each of s,
t, u and v is 1; [0075] (ii) R.sup.a, R.sup.b, R.sup.c, R.sup.d,
R.sup.e, R.sup.f, R.sup.g and R.sup.h is hydrogen, s is 0, and each
of t, u and v is 1; [0076] (iii) R.sup.a, R.sup.b, R.sup.c,
R.sup.d, R.sup.e, R.sup.f, R.sup.g and R.sup.h is hydrogen, s is 0,
each of t and v is 1 and u is 2; or [0077] (iv) R.sup.a, R.sup.b,
R.sup.c, R.sup.d, R.sup.e, R.sup.f, R.sup.g and R.sup.h is
hydrogen, s is 0, t is 1 and the sum of u and v is 1, 2 or 3.
[0078] Further preferred is a compound having the formula (I), (II)
or (III), wherein [0079] (i) each of R.sup.a, R.sup.b, R.sup.c,
R.sup.d, R.sup.e, R.sup.f, R.sup.g and R.sup.h is hydrogen, s is 0,
t is 1 and u+v=3; [0080] (ii) each of R.sup.a, R.sup.b, R.sup.c,
R.sup.d, R.sup.e, R.sup.f, R.sup.g and R.sup.h is hydrogen and s,
t, u and v are each 1; [0081] (iii) each of R.sup.a, R.sup.b,
R.sup.c, R.sup.d, R.sup.e, R.sup.f, R.sup.g and R.sup.h is
hydrogen, s is 0 and t, u and v are each 1; [0082] (iv) each of
R.sup.a, R.sup.b, R.sup.c, R.sup.d, R.sup.e, R.sup.f, R.sup.g and
R.sup.h is hydrogen and the sum of s, t, u and v is 2; or [0083]
(v) each of R.sup.a, R.sup.b, R.sup.c, R.sup.d, R.sup.e, R.sup.f,
R.sup.g and R.sup.h is hydrogen and the sum of s, t, u and v is
1.
[0084] Further preferred is a compound having the formula (I), (II)
or (III), wherein D and E are independently selected from CH and
N.
[0085] Further preferred is a compound having the formula (I), (II)
or (III), wherein D is CH and E is N.
[0086] Further preferred is a compound having the formula (I), (II)
or (III), wherein D is N and E is CH.
[0087] Further preferred is a compound having the formula (I), (II)
or (III), wherein L (i.e., L.sub.1-L.sub.2-L.sub.3) is selected
from
##STR00014##
[0088] Further preferred is a compound having the formula (I), (II)
or (III) wherein L (i.e., L.sub.1-L.sub.2-L.sub.3) is selected
from
##STR00015##
[0089] Further preferred is a compound having the formula (I), (II)
or (III), wherein (i.e., L.sub.1-L.sub.2-L.sub.3) is selected
from
##STR00016##
[0090] Further preferred is a compound having the formula (I), (II)
or (III), wherein L.sub.2 is selected from
##STR00017##
[0091] Further preferred is a compound having the formula (I), (II)
or (III), wherein L.sub.2 is selected from
##STR00018##
[0092] Further preferred is a compound having the formula (I), (II)
or (III), wherein L.sub.1 and L.sub.3 are independently absent or
--CH.sub.2--.
[0093] Further preferred is a compound having the formula (I), (II)
or (III), wherein [0094] (i) L.sub.1 and L.sub.3 are absent; [0095]
(ii) L.sub.1 and L.sub.3 are CH.sub.2--; [0096] (iii) L.sub.1 is
absent and L.sub.3 is --CH.sub.2--; or [0097] (iv) L.sub.1 is
--CH.sub.2-- and L.sub.3 is absent.
[0098] Further preferred is a compound having the formula (I), (II)
or (III), wherein each occurrence of R.sup.3 is independently
hydrogen, halogen or substituted or unsubstituted C.sub.1-3
alkyl.
[0099] Further preferred is a compound having the formula (I), (II)
or (III), wherein each occurrence of R.sup.3 is independently
hydrogen, fluorine or methyl.
[0100] Further preferred is a compound having the formula (I), (II)
or (III), wherein at least one of R.sup.1 or R.sup.2 is
hydrogen.
[0101] Further preferred is a compound having the formula (I), (II)
or (III), wherein each of R.sup.1 and R.sup.2 are independently
selected from halogen, substituted or unsubstituted alkyl,
--NR.sup.zR.sup.z, substituted or unsubstituted heterocyclyl,
substituted or unsubstituted heterocyclylalkyl, substituted or
unsubstituted aryl, substituted or unsubstituted arylalkyl,
substituted or unsubstituted heteroaryl, and substituted or
unsubstituted heteroarylalkyl.
[0102] Further preferred is a compound having the formula (I), (II)
or (III), wherein each of R.sup.1 and R.sup.2 is independently
selected from --NR.sup.zR.sup.z, substituted or unsubstituted aryl
and substituted or unsubstituted heteroaryl.
[0103] Further preferred is a compound having the formula (I), (II)
or (III), wherein each of R.sup.1 and R.sup.2 is independently
substituted or unsubstituted aryl.
[0104] Further preferred is a compound having the formula (I), (II)
or (III), wherein each of R.sup.1 and R.sup.2 is independently
substituted or unsubstituted heteroaryl.
[0105] Further preferred is a compound having the formula (I), (II)
or (III), wherein [0106] (i) R.sup.1 is substituted or
unsubstituted aryl and R.sup.2 is substituted or unsubstituted
heteroaryl; [0107] (ii) R.sup.1 is substituted or unsubstituted
heteroaryl and R.sup.2 is substituted or unsubstituted aryl; [0108]
(iii) both R.sup.1 and R.sup.2 are, independently, substituted or
unsubstituted aryl; or [0109] (iv) both R.sup.1 and R.sup.2 are,
independently, substituted or unsubstituted heteroaryl.
[0110] Further preferred is a compound having the formula (I), (II)
or (III), wherein each of R.sup.1 and R.sup.2 is independently
selected from
##STR00019## ##STR00020##
[0111] Further preferred is a compound having the formula (I), (II)
or (III), wherein each of R.sup.x and R.sup.y is independently
selected from hydrogen, hydroxyl or --CH.sub.2OH.
[0112] Further preferred is a compound having the formula (I), (II)
or (III), wherein each of R.sup.x and R.sup.y is hydrogen.
[0113] Representative compounds of the present invention include
those listed below (see also Table 1) and pharmaceutically
acceptable salts thereof. The present invention should not be
construed to be limited to these compounds. [0114] 1.
2-(Pyridin-2-yl)-N-(5-(1-(6-(2-(3-(trifluoromethoxy)phenyl)acetamido)pyri-
din-3-yl) piperidin-4-yl)-1,3,4-thiadiazol-2-yl)acetamide; [0115]
2.
2-(Pyridin-2-yl)-N-(5-(1-(6-(2-(3-(trifluoromethoxy)phenyl)acetamido)pyri-
din-3-yl) piperidin-3-yl)-1,3,4-thiadiazol-2-yl)acetamide; [0116]
2A. (R) or (S)
2-(Pyridin-2-yl)-N-(5-(1-(6-(2-(3-(trifluoromethoxy)phenyl)acetami-
do) pyridin-3-yl)piperidin-3-yl)-1,3,4-thiadiazol-2-yl)acetamide;
[0117] 2B. (S) or (R)
2-(Pyridin-2-yl)-N-(5-(1-(6-(2-(3-(trifluoromethoxy)phenyl)acetamido)
pyridin-3-yl)piperidin-3-yl)-1,3,4-thiadiazol-2-yl)acetamide;
[0118] 3.
2-(Pyridin-3-yl)-N-(5-(1-(6-(2-(3-(trifluoromethoxy)phenyl)acetamido)pyri-
din-3-yl) piperidin-3-yl)-1,3,4-thiadiazol-2-yl)acetamide; [0119]
3A. (R) or (S)
2-(Pyridin-3-yl)-N-(5-(1-(6-(2-(3-(trifluoromethoxy)phenyl)acetami-
do) pyridin-3-yl)piperidin-3-yl)-1,3,4-thiadiazol-2-yl)acetamide;
[0120] 3B. (S) or (R)
2-(Pyridin-3-yl)-N-(5-(1-(6-(2-(3-(trifluoromethoxy)phenyl)acetamido)
pyridin-3-yl)piperidin-3-yl)-1,3,4-thiadiazol-2-yl)acetamide;
[0121] 4.
2-(Pyridin-3-yl)-N-(5-(1-(6-(2-(3-(trifluoromethoxy)phenyl)acetamido)pyri-
din-3-yl) piperidin-4-yl)-1,3,4-thiadiazol-2-yl)acetamide; [0122]
5.
2-(3-Cyanophenyl)-N-(5-(1-(6-(2-(3-(trifluoromethoxy)phenyl)acetamido)pyr-
idin-3-yl)piperidin-4-yl)-1,3,4-thiadiazol-2-yl)acetamide; [0123]
6.
2-(Pyridin-2-yl)-N-(5-(4-(5-(2-(3-(trifluoromethoxy)phenyl)acetamido)-1,3-
,4-thiadiazol-2-yl)piperidin-1-yl)pyridin-2-yl)acetamide; [0124] 7.
2-(Pyridin-2-yl)-N-(5-(3-(5-(2-(3-(trifluoromethoxy)phenyl)acetanido)-1,3-
,4-thiadiazol-2-yl)piperidin-1-yl)pyridin-2-yl)acetamide; [0125] 8.
2-(pyridin-2-yl)-N-(5-(1-(6-(2-(3-(trifluoromethoxy)phenyl)acetamido)pyri-
dazin-3-yl) piperidin-4-yl)-1,3,4-thiadiazol-2-yl)acetamide; [0126]
9.
2-(Pyridin-2-yl)-N-(5-(1-(6-(2-(3-(trifluoromethoxy)phenyl)acetamido)pyri-
dazin-3-yl)piperidin-3-yl)-1,3,4-thiadiazol-2-yl)acetamide; [0127]
10.
2-(Pyridin-3-yl)-N-(5-(1-(6-(2-(3-(trifluoromethoxy)phenyl)acetamido)pyri-
dazin-3-yl)piperidin-4-yl)-1,3,4-thiadiazol-2-yl)acetamide; [0128]
11.
2-(3-(Methylsulfonamido)phenyl)-N-(5-(1-(6-(2-(3-(trifluoromethoxy)phenyl-
)
acetamido)pyridazin-3-yl)piperidin-4-yl)-1,3,4-thiadiazol-2-yl)acetamide-
; [0129] 12.
2-(2-Chlorophenyl)-N-(6-(4-(5-(2-(pyridin-2-yl)acetamido)-1,3,4-thiadiazo-
l-2-yl) piperidin-1-yl)pyridazin-3-yl)acetamide; [0130] 13.
2-(2-Chlorophenyl)-N-(5-(1-(6-(2-(3-(trifluoromethoxy)phenyl)acetamido)py-
ridazin-3-yl)piperidin-3-yl)-1,3,4-thiadiazol-2-yl)acetamide;
[0131] 14.
2-(2-Fluorophenyl)-N-(6-(4-(5-(2-(pyridin-2-yl)acetamido)-1,3,4-thiadiazo-
l-2-yl) piperidin-1-yl)pyridazin-3-yl)acetamide; [0132] 15.
2-(Pyrazin-2-yl)-N-(5-(1-(6-(2-(3-(trifluoromethoxy)phenyl)acetamido)pyri-
dazin-3-yl)piperidin-4-yl)-1,3,4-thiadiazol-2-yl)acetamide; [0133]
16.
2-(Pyridin-2-yl)-N-(5-(1-(6-(2-(3-(trifluoromethoxy)phenyl)acetamido)pyri-
dazin-3-yl)piperidin-4-yl)-1,3,4-thiadiazol-2-yl)acetamide
dihydrochloride; [0134] 17.
2-(Pyridin-2-yl)-N-(6-(4-(5-(2-(3-(trifluoromethoxy)phenyl)acetamido)-1,3-
,4-thiadiazol-2-yl)piperidin-1-yl)pyridazin-3-yl)acetamide; [0135]
18.
2-(Pyridin-3-yl)-N-(6-(4-(5-(2-(3-(trifluoromethoxy)phenyl)acetamido)-1,3-
,4-thiadiazol-2-yl)piperidin-1-yl)pyridazin-3-yl)acetamide; [0136]
19.
2-(Pyridin-3-yl)-N-(6-(4-(5-(2-(2,3,6-trifluorophenyl)acetamido)-1,3,4-th-
iadiazol-2-yl)piperidin-1-yl)pyridazin-3-yl)acetamide; [0137] 20.
2-(Pyridin-2-yl)-N-(6-(4-(5-(2-(2,3,6-trifluorophenyl)acetamido)-1,3,4-th-
iadiazol-2-yl)piperidin-1-yl)pyridazin-3-yl)acetamide; [0138] 21.
2-(2,3-Difluorophenyl)-N-(5-(1-(6-(2-(pyridin-2-yl)acetamido)pyridazin-3--
yl)piperidin-4-yl)-1,3,4-thiadiazol-2-yl)acetamide; [0139] 22.
2-(3,4-Difluorophenyl)-N-(5-(1-(6-(2-(pyridin-2-yl)acetamido)pyridazin-3--
yl)piperidin-4-yl)-1,3,4-thiadiazol-2-yl)acetamide; [0140] 23.
2-(2-Fluorophenyl)-N-(5-(1-(6-(2-(pyridin-2-yl)acetamido)pyridazin-3-yl)p-
iperidin-4-yl)-1,3,4-thiadiazol-2-yl)acetamide; [0141] 24.
2-(3-Fluorophenyl)-N-(5-(1-(6-(2-(pyridin-2-yl)acetamido)pyridazin-3-yl)p-
iperidin-4-yl)-1,3,4-thiadiazol-2-yl)acetamide; [0142] 25.
2-(4-Fluorophenyl)-N-(5-(1-(6-(2-(pyridin-2-yl)acetamido)pyridazin-3-yl)p-
iperidin-4-yl)-1,3,4-thiadiazol-2-yl)acetamide; [0143] 26.
2-(2-Methoxyphenyl)-N-(5-(1-(6-(2-(pyridin-2-yl)acetamido)pyridazin-3-yl)-
piperidin-4-yl)-1,3,4-thiadiazol-2-yl)acetamide; [0144] 27.
2-(2-Chlorophenyl)-N-(5-(1-(6-(2-(pyridin-2-yl)acetamido)pyridazin-3-yl)p-
iperidin-4-yl)-1,3,4-thiadiazol-2-yl)acetamide; [0145] 28.
2-(5-Chloro-2-(trifluoromethyl)phenyl)-N-(5-(1-(6-(2-(pyridin-2-yl)acetam-
ido)
pyridazin-3-yl)piperidin-4-yl)-1,3,4-thiadiazol-2-yl)acetamide;
[0146] 29.
2-(4-Chlorophenyl)-N-(5-(1-(6-(2-(pyridin-2-yl)acetamido)pyridazin-3-yl)p-
iperidin-4-yl)-1,3,4-thiadiazol-2-yl)acetamide; [0147] 30.
2-(Quinolin-6-yl)-N-(5-(1-(6-(2-(3-(trifluoromethoxy)phenyl)acetamido)pyr-
idazin-3-yl)piperidin-4-yl)-1,3,4-thiadiazol-2-yl)acetamide; [0148]
31.
2-o-Tolyl-N-(5-(1-(6-(2-(3-(trifluoromethoxy)phenyl)acetamido)pyridazin-3-
-yl) piperidin-4-yl)-1,3,4-thiadiazol-2-yl)acetamide; [0149] 32.
N-(6-(4-(5-(2-(1H-indol-3-yl)acetamido)-1,3,4-thiadiazol-2-yl)piperidin-1-
-yl) pyridazin-3-yl)-2-(3-(trifluoromethoxy)phenyl)acetamide;
[0150] 33.
2-(2-Fluorophenyl)-N-(6-(4-(5-(2-(pyrazin-2-yl)acetamido)-1,3,4-thiadiazo-
l-2-yl) piperidin-1-yl)pyridazin-3-yl)acetamide; [0151] 34.
2-(3-(Azetidin-1-yl)phenyl)-N-(5-(1-(6-(2-(3-(trifluoromethoxy)phenyl)ace-
tamido)
pyridazin-3-yl)piperidin-4-yl)-1,3,4-thiadiazol-2-yl)acetamide;
[0152] 35.
2-(3-Chlorophenyl)-N-(5-(1-(6-(2-(pyridin-2-yl)acetamido)pyridazin-3-yl)p-
iperidin-4-yl)-1,3,4-thiadiazol-2-yl)acetamide; [0153] 36.
3-Hydroxy-2-phenyl-N-(5-(1-(6-(2-(3-(trifluoromethoxy)phenyl)acetamido)
pyridazin-3-yl)piperidin-4-yl)-1,3,4-thiadiazol-2-yl)propanamide;
[0154] 37.
(R)-2-hydroxy-2-phenyl-N-(5-(1-(6-(2-(3-(trifluoromethoxy)phenyl)acet-
amido)
pyridazin-3-yl)piperidin-4-yl)-1,3,4-thiadiazol-2-yl)acetamide;
[0155] 38.
2-(3-(3-Fluoroazetidin-1-yl)phenyl)-N-(5-(1-(6-(2-(3-(trifluoromethoxy)ph-
enyl)
acetamido)pyridazin-3-yl)piperidin-4-yl)-1,3,4-thiadiazol-2-yl)aceta-
mide; [0156] 39.
2-(Pyridin-2-yl)-N-(5-((1-(6-(2-(3-(trifluoromethoxy)phenyl)acetamido)pyr-
idazin-3-yl)
piperidin-4-yl)methyl)-1,3,4-thiadiazol-2-yl)acetamide; or a
pharmaceutically acceptable salt thereof.
TABLE-US-00002 [0156] TABLE 1 Ex. Structure 1 ##STR00021## 2
##STR00022## 2A ##STR00023## 2B ##STR00024## 3 ##STR00025## 3A
##STR00026## 3B ##STR00027## 4 ##STR00028## 5 ##STR00029## 6
##STR00030## 7 ##STR00031## 8 ##STR00032## 9 ##STR00033## 10
##STR00034## 11 ##STR00035## 12 ##STR00036## 13 ##STR00037## 14
##STR00038## 15 ##STR00039## 16 ##STR00040## 17 ##STR00041## 18
##STR00042## 19 ##STR00043## 20 ##STR00044## 21 ##STR00045## 22
##STR00046## 23 ##STR00047## 24 ##STR00048## 25 ##STR00049## 26
##STR00050## 27 ##STR00051## 28 ##STR00052## 29 ##STR00053## 30
##STR00054## 31 ##STR00055## 32 ##STR00056## 33 ##STR00057## 34
##STR00058## 35 ##STR00059## 36 ##STR00060## 37 ##STR00061## 38
##STR00062## 39 ##STR00063##
[0157] Yet another embodiment of the present invention is a method
for inhibiting glutaminase in a patient by administering to the
patient an effective amount of at least one compound of the present
invention (for example, a compound of formula (I), (II) or (III) as
defined above).
[0158] Yet another embodiment of the present invention is a method
for treating an inflammatory, autoimmune or proliferative disease
(e.g., via inhibition of glutaminase) by administering to a patient
in need of such treatment an effective amount of at least one
compound of the present invention. In one embodiment, the compound
of the present invention inhibits glutaminase (i.e., an effective
amount of the compound is administered to inhibit glutaminase).
[0159] Yet another embodiment of the present invention is a method
for treating a inflammatory, autoimmune or proliferative disease
(e.g., via inhibition of glutaminase) by administering to a patient
in need of such treatment an effective amount of at least one
compound of the present invention, in combination (simultaneously
or sequentially) with at least one other anti-inflammatory,
immunomodulator or anti-cancer agent. In one embodiment, the
compound of the present invention inhibits glutaminase.
[0160] More particularly, the compounds of formula (I) to (III) and
pharmaceutically acceptable esters or salts thereof can be
administered for the treatment, prevention and/or amelioration of
diseases or disorders associated with glutamine, in particular the
amelioration of diseases or disorders mediated by glutamine,
including, but not limited to, inflammatory diseases or disorders,
autoimmune diseases or disorders, and cancer and other
proliferative diseases or disorders.
[0161] The compounds of the present invention are useful in the
treatment of a variety of cancers, including, but not limited to:
[0162] carcinoma, including that of the bladder, breast, colon,
kidney, liver, lung, including small cell lung cancer, esophagus,
gall bladder, ovary, pancreas, stomach, cervix, thyroid, prostate,
and skin, including squamous cell carcinoma; [0163] hematopoietic
tumors of lymphoid lineage, including leukemia, acute lymphocytic
leukemia, acute lymphoblastic leukemia, B-cell lymphoma, T-cell
lymphoma, Hodgkin's lymphoma, non-Hodgkins lymphoma, hairy cell
lymphoma and Burkett's lymphoma; [0164] hematopoietic tumors of
myeloid lineage, including acute and chronic myelogenous leukemias,
myelodysplastic syndrome and promyelocytic leukemia; [0165] tumors
of mesenchymal origin, including fibrosarcoma and rhabdomyosarcoma;
[0166] tumors of the central and peripheral nervous system,
including astrocytoma, neuroblastoma, glioma and schwannomas; and
[0167] other tumors, including melanoma, seminoma, teratocarcinoma,
osteosarcoma, xenoderoma pigmentosum, keratoctanthoma, thyroid
follicular cancer and Kaposi's sarcoma.
[0168] Due to the key role of glutaminase and glutamine in the
regulation of cellular proliferation, the glutaminase inhibitors of
the present invention may act as reversible cytostatic agents and
therefore may be useful in the treatment of any disease process
which features abnormal cellular proliferation, e.g., benign
prostatic hyperplasia, familial adenomatosis polyposis,
neuro-fibromatosis, atherosclerosis, pulmonary fibrosis, arthritic
disease (e.g., arthritis), psoriasis, glomerulonephritis,
restenosis following angioplasty or vascular surgery, hypertrophic
scar formation, inflammatory bowel disease, transplantation
rejection, endotoxic shock, and fungal infections.
[0169] The compounds of the present invention as modulators of
apoptosis are useful in the treatment of cancer (including but not
limited to those types mentioned herein above), viral infections
(including but not limited to herpevirus, poxvirus, Epstein-Barr
virus, Sindbis virus and adenovirus), prevention of AIDS
development in HIV-infected individuals, autoimmune diseases
(including but not limited to systemic lupus, erythematosus,
autoimmune mediated glomerulonephritis, rheumatoid arthritis,
psoriasis, inflammatory bowel disease, and autoimmune diabetes
mellitus), neurodegenerative disorders (including but not limited
to Alzheimer's disease, AIDS-related dementia, Parkinson's disease,
amyotrophic lateral sclerosis, retinitis pigmentosa, spinal
muscular atrophy and cerebellar degeneration), myelodysplastic
syndromes, aplastic anemia, ischemic injury associated with
myocardial infarctions, stroke and reperfusion injury, arrhythmia,
atherosclerosis, toxin-induced or alcohol related liver diseases,
hematological diseases (including but not limited to chronic anemia
and aplastic anemia), degenerative diseases of the musculoskeletal
system (including but not limited to osteoporosis and arthritis)
aspirin-sensitive rhinosinusitis, cystic fibrosis, multiple
sclerosis, kidney diseases and cancer pain.
[0170] The compounds of present invention can modulate the level of
cellular RNA and DNA synthesis. These agents are therefore useful
in the treatment of viral infections (including but not limited to
HIV, human papilloma virus, herpesvirus, poxvirus, Epstein-Barr
virus, Sindbis virus and adenovirus).
[0171] The compounds of the present invention are useful in the
chemoprevention of cancer. Chemoprevention is defined as inhibiting
the development of invasive cancer by either blocking the
initiating mutagenic event or by blocking the progression of
pre-malignant cells that have already suffered an insult or
inhibiting tumor relapse. The compounds described herein are also
useful in inhibiting tumor angiogenesis and metastasis. One
embodiment of the invention is a method of inhibiting tumor
angiogenesis or metastasis in a patient in need thereof by
administering an effective amount of one or more compounds of the
present invention.
[0172] Another embodiment of the present invention is a method of
treating an immune system-related disease (e.g., an autoimmune
disease), a disease or disorder involving inflammation (e.g.,
asthma, chronic obstructive pulmonary disease, rheumatoid
arthritis, inflammatory bowel disease, glomerulonephritis,
neuroinflammatory diseases, multiple sclerosis, uveitis and
disorders of the immune system), cancer or other proliferative
disease, a hepatic disease or disorder, or a renal disease or
disorder. The method includes administering an effective amount of
one or more compounds of the present invention.
[0173] Examples of immune disorders include, but are not limited
to, psoriasis, rheumatoid arthritis, vasculitis, inflammatory bowel
disease, dermatitis, osteoarthritis, asthma, inflammatory muscle
disease, allergic disease (e.g., allergic rhinitis), vaginitis,
interstitial cystitis, scleroderma, osteoporosis, eczema,
allogeneic or xenogeneic transplantation (organ, bone marrow, stem
cells and other cells and tissues) graft rejection,
graft-versus-host disease, lupus erythematosus, inflammatory
disease, type I diabetes, pulmonary fibrosis, dermatomyositis,
Sjogren's syndrome, thyroiditis (e.g., Hashimoto's and autoimmune
thyroiditis), myasthenia gravis, autoimmune hemolytic anemia,
multiple sclerosis, cystic fibrosis, chronic relapsing hepatitis,
primary biliary cirrhosis, allergic conjunctivitis and atopic
dermatitis.
[0174] In one embodiment, the compounds described herein are used
as immunosuppressants to prevent transplant graft rejections,
allogeneic or xenogeneic transplantation rejection (organ, bone
marrow, stem cells, other cells and tissues), and
graft--versus--host disease. In other embodiments, transplant graft
rejections result from tissue or organ transplants. In further
embodiments, graft-versus-host disease results from bone marrow or
stem cell transplantation. One embodiment is a method of preventing
or decreasing the risk of transplant graft rejection, allogeneic or
xenogeneic transplantation rejection (organ, bone marrow, stem
cells, other cells and tissues), or graft--versus--host disease by
administering an effective amount of one or more compounds of the
present invention.
[0175] The compounds of the present invention are also useful in
combination (administered together or sequentially) with known
anti-cancer treatments, such as radiation therapy or with
cytostatic, cytotoxic or anticancer agents, such as for example,
but not limited to, DNA interactive agents, such as cisplatin or
doxorubicin; topoisomerase II inhibitors, such as etoposide;
topoisomerase I inhibitors such as CPT-11 or topotecan; tubulin
interacting agents, such as paclitaxel, docetaxel or the
epothilones (for example ixabepilone), either naturally occurring
or synthetic; hormonal agents, such as tamoxifen; thymidilate
synthase inhibitors, such as 5-fluorouracil; and anti-metabolites,
such as methotrexate, other tyrosine kinase inhibitors such as
Iressa and OSI-774; angiogenesis inhibitors; EGF inhibitors; VEGF
inhibitors; CDK inhibitors; SRC inhibitors; c-Kit inhibitors;
Her1/2 inhibitors and monoclonal antibodies directed against growth
factor receptors such as erbitux (EGF) and herceptin (Her2) and
other protein kinase modulators as well.
[0176] The compounds of the present invention are also useful in
combination (administered together or sequentially) with one or
more steroidal anti-inflammatory drugs, non-steroidal
anti-inflammatory drugs (NSAIDs) or immune selective
anti-inflammatory derivatives (ImSAIDs).
[0177] The invention further provides a pharmaceutical composition
comprising one or more compounds of the present invention (such as
a compound having formula (I), (II) or (III)) together with a
pharmaceutically acceptable carrier. The pharmaceutical composition
may further comprise one or more of the active ingredients
identified above, such as other steroidal anti-inflammatory drugs,
non-steroidal anti-inflammatory drugs (NSAIDs), immune selective
anti-inflammatory derivatives (ImSAIDs) or anti-cancer agents.
[0178] In one embodiment, the pharmaceutical composition includes a
therapeutically effective amount of one or more compounds of
formula (I), (II) or (III).
[0179] Yet another embodiment is a method of treating autoimnune
disorders in a patient in need thereof by administering a
therapeutically effective amount of a compound of the present
invention. For example, the compounds of the present invention are
effective for treating asthma, chronic obstructive pulmonary
disease (COPD), rheumatoid arthritis, psoriasis, lupus and
experimental autoimmune encephalomyelitis (EAE).
[0180] Yet another embodiment is a method of treating allergic
rhinitis in a patient in need thereof by administering a
therapeutically effective amount of a compound of the present
invention.
[0181] Yet another embodiment is a method of treating cancer in a
patient in need thereof by administering a therapeutically
effective amount of a compound of the present invention. For
example, the compounds of the present invention are effective for
treating hematopoietic tumors of lymphoid lineage, leukemia, acute
lymphocytic leukemia, acute lymphoblastic leukemia, B-cell
lymphoma, T-cell lymphoma, Hodgkin's lymphoma, non-Hodgkins
lymphoma, hairy cell lymphoma and Burkett's lymphoma; hematopoietic
tumors of myeloid lineage, acute myelogenous leukemias, chronic
myelogenous leukemias, myelodysplastic syndrome and promyelocytic
leukemia. The compounds of the present invention are also effective
for treating carcinoma of the bladder, carcinoma of the breast,
carcinoma of the colon, carcinoma of the kidney, carcinoma of the
liver, carcinoma of the lung, small cell lung cancer, esophageal
cancer, gall bladdercancer, ovarian cancer, pancreatic cancer,
stomach cancer, cervical cancer, thyroid cancer, prostate cancer,
skin cancer, squamous cell carcinoma, tumors of mesenchymal origin,
fibrosarcoma, rhabdomyosarcoma, tumors of the central and
peripheral nervous system, astrocytoma, neuroblastoma, glioma,
schwannoma, melanoma, seminoma, teratocarcinoma, osteosarcoma,
xenoderoma pigmentosum, keratoctanthoma, thyroid follicular cancer
and Kaposi's sarcoma.
[0182] Yet another embodiment is a method of treating leukemia in a
patient in need thereof by administering a therapeutically
effective amount of a compound of the present invention. For
example, the compounds of the present invention are effective for
treating chronic lymphocytic leukemia (CLL), non-Hodgkin lymphoma
(NHL), acute myeloid leukemia (AML), acute lymphoblastic leukemia
(ALL), multiple myeloma (MM), small lymphocytic lymphoma (SLL), and
indolent non-Hodgkin's lymphoma (I-NHL).
DETAILED DESCRIPTION OF THE INVENTION
[0183] As used herein the following definitions shall apply unless
otherwise indicated. Further many of the groups defined herein can
be optionally substituted. The listing of substituents in the
definition is exemplary and is not to be construed to limit the
substituents defined elsewhere in the specification.
[0184] The term "alkyl", unless otherwise specified, refers to a
straight or branched hydrocarbon chain radical consisting solely of
carbon and hydrogen atoms, containing no unsaturation, having from
one to eight carbon atoms, and which is attached to the rest of the
molecule by a single bond, e.g., methyl, ethyl, n-propyl,
1-methylethyl (isopropyl), n-butyl, n-pentyl, and 1,1-dimethylethyl
(t-butyl). The term "C.sub.1-6 alkyl" refers to an alkyl group as
defined above having up to 6 carbon atoms. The term "C.sub.1-3
alkyl" refers to an alkyl group as defined above having up to 3
carbon atoms. In appropriate circumstances, the term "alkyl" refers
to a hydrocarbon chain radical as mentioned above which is
bivalent.
[0185] The term "alkenyl", unless otherwise specified, refers to an
aliphatic hydrocarbon group containing one or more carbon-carbon
double bonds and which may be a straight or branched or branched
chain having about 2 to about 10 carbon atoms, e.g., ethenyl,
1-propenyl, 2-propenyl (allyl), iso-propenyl, 2-methyl-1-propenyl,
1-butenyl, and 2-butenyl. The term "C.sub.2-6 alkenyl" refers to an
alkenyl group as defined above having up to 6 carbon atoms. In
appropriate circumstances, the term "alkenyl" refers to a
hydrocarbon group as mentioned above which is bivalent.
[0186] The term "alkynyl", unless otherwise specified, refers to a
straight or branched chain hydrocarbyl radical having at least one
carbon-carbon triple bond, and having in the range of 2 to up to 12
carbon atoms (with radicals having in the range of 2 to up to 10
carbon atoms presently being preferred) e.g., ethynyl, propynyl,
and butnyl. The term "C.sub.2-6 alkynyl" refers to an alkynyl group
as defined above having up to 6 carbon atoms. In appropriate
circumstances, the term "alkynyl" refers to a hydrocarbyl radical
as mentioned above which is bivalent.
[0187] The term "alkoxy" unless otherwise specified, denotes an
alkyl, cycloalkyl, or cycloalkylalkyl group as defined above
attached via an oxygen linkage to the rest of the molecule. The
term "substituted alkoxy" refers to an alkoxy group where the alkyl
constituent is substituted (i.e., --O-(substituted alkyl). For
example "alkoxy" refers to the group --O-alkyl, including from 1 to
8 carbon atoms of a straight, branched, cyclic configuration and
combinations thereof attached to the parent structure through an
oxygen atom. Examples include methoxy, ethoxy, propoxy, isopropoxy,
cyclopropyloxy, and cyclohexyloxy. In appropriate circumstances,
the term "alkoxy" refers to a group as mentioned above which is
bivalent.
[0188] The term "cycloalkyl", unless otherwise specified, denotes a
non-aromatic mono or multicyclic ring system of about 3 to 12
carbon atoms such as cyclopropyl, cyclobutyl, cyclopentyl, and
cyclohexyl. Examples of multicyclic cycloalkyl groups include
perhydronaphthyl, adamantyl and norbornyl groups, bridged cyclic
groups, and sprirobicyclic groups, e.g., sprio (4,4) non-2-yl. The
term "C.sub.3-6 cycloalkyl" refers to a cycloalkyl group as defined
above having up to 6 carbon atoms.
[0189] The term "cycloalkylalkyl", unless otherwise specified,
refers to a cyclic ring-containing radical containing in the range
of about 3 up to 8 carbon atoms directly attached to an alkyl group
which is then attached to the main structure at any carbon from the
alkyl group, such as cyclopropylmethyl, cyclobutylethyl, and
cyclopentylethyl.
[0190] The term "cycloalkenyl", unless otherwise specified, refers
to cyclic ring-containing radicals containing in the range of about
3 up to 8 carbon atoms with at least one carbon-carbon double bond
such as cyclopropenyl, cyclobutenyl, and cyclopentenyl. The term
"cycloalkenylalkyl" refers to a cycloalkenyl group directly
attached to an alkyl group which is then attached to the main
structure at any carbon from the alkyl group.
[0191] The term "aryl", unless otherwise specified, refers to
aromatic radicals having in the range of 6 up to 20 carbon atoms
such as phenyl, naphthyl, tetrahydronaphthyl, indanyl, and
biphenyl.
[0192] The term "arylalkyl", unless otherwise specified, refers to
an aryl group as defined above directly bonded to an alkyl group as
defined above, e.g., --CH.sub.2C.sub.6H.sub.5 and
--C.sub.2H.sub.5C.sub.6H.sub.5.
[0193] The term "heterocyclic ring", unless otherwise specified,
refers to a non-aromatic 3 to 15 member ring radical which consists
of carbon atoms and at least one heteroatom selected from nitrogen,
phosphorus, oxygen and sulfur. For purposes of this invention, the
heterocyclic ring radical may be a mono-, bi-, tri- or tetracyclic
ring system, which may include fused, bridged or spiro ring
systems, and the nitrogen, phosphorus, carbon, oxygen or sulfur
atoms in the heterocyclic ring radical may be optionally oxidized
to various oxidation states. In addition, the nitrogen atom may be
optionally quaternized. The heterocyclic ring radical may be
attached to the main structure at any heteroatom or carbon
atom.
[0194] The term "heterocyclyl", unless otherwise specified, refers
to a heterocylic ring radical as defined above. The heterocylcyl
ring radical may be attached to the main structure at any
heteroatom or carbon atom. In appropriate circumstances, the term
"heterocyclyl" refers to a hydrocarbon chain radical as mentioned
above which is bivalent.
[0195] The term "heterocyclylalkyl", unless otherwise specified,
refers to a heterocylic ring radical as defined above directly
bonded to an alkyl group. The heterocyclylalkyl radical may be
attached to the main structure at any carbon atom in the alkyl
group. Examples of such heterocycloalkyl radicals include, but are
not limited to, dioxolanyl, thienyl[1,3]dithianyl,
decahydroisoquinolyl, imidazolinyl, imidazolidinyl,
isothiazolidinyl, isoxazolidinyl, morpholinyl, octahydroindolyl,
octahydroisoindolyl, 2-oxopiperazinyl, 2-oxopiperidinyl,
2-oxopyrrolidinyl, oxazolidinyl, piperidinyl, piperazinyl,
4-piperidonyl, pyrrolidinyl, pyrazolidinyl, quinuclidinyl,
thiazolidinyl, tetrahydrofuryl, trithianyl, tetrahydropyranyl,
thiomorpholinyl, thiamorpholinyl, 1-oxo-thiomorpholinyl, and
1,1-dioxo-thiomorpholinyl.
[0196] The term "heteroaryl", unless otherwise specified, refers to
an optionally substituted 5 to 14 member aromatic ring having one
or more heteroatoms selected from N, O, and S as ring atoms. The
heteroaryl may be a mono-, bi- or tricyclic ring system. Examples
of such "heterocyclic ring" or "heteroaryl" radicals include, but
are not limited to, oxazolyl, thiazolyl, imidazolyl, pyrrolyl,
furanyl, pyridinyl, pyrimidinyl, pyrazinyl, benzofuranyl, indolyl,
benzothiazolyl, benzoxazolyl, carbazolyl, quinolyl, isoquinolyl,
azetidinyl, acridinyl, benzodioxolyl, benzodioxanyl, benzofuranyl,
carbazolyl, cinnolinyl, dioxolanyl, indolizinyl, naphthyridinyl,
perhydroazepinyl, phenazinyl, phenothiazinyl, phenoxazinyl,
phthalazinyl, pteridinyl, purinyl, quinazolinyl, quinoxalinyl,
tetrazoyl, tetrahydroisoquinolyl, piperidinyl, piperazinyl,
2-oxopiperazinyl, 2-oxopiperidinyl, 2-oxopyrrolidinyl,
2-oxoazepinyl, azepinyl, 4-piperidonyl, pyrrolidinyl, pyridazinyl,
oxazolinyl, oxazolidinyl, triazolyl, indanyl, isoxazolyl,
isoxazolidinyl, morpholinyl, thiazolinyl, thiazolidinyl,
isothiazolyl, quinuclidinyl, isothiazolidinyl, isoindolyl,
indolinyl, isoindolinyl, octahydroindolyl, octahydroisoindolyl,
decahydroisoquinolyl, benzimidazolyl, thiadiazolyl, benzopyranyl,
tetrahydrofuryl, tetrahydropyranyl, thienyl, benzothienyl,
thiamorpholinyl, thiamorpholinyl sulfoxide, thiamorpholinyl
sulfone, dioxaphospholanyl, oxadiazolyl, chromanyl, and
isochromanyl. The heteroaryl ring radical may be attached to the
main structure at any heteroatom or carbon atom. The term
"substituted heteroaryl" also includes ring systems substituted
with one or more oxide (--O--) substituents, such as pyridinyl
N-oxides.
[0197] The term "heteroarylalkyl", unless otherwise specified,
refers to a heteroaryl ring radical as defined above directly
bonded to an alkyl group. The heteroarylalkyl radical may be
attached to the main structure at any carbon atom from alkyl
group.
[0198] The term "cyclic ring" refers to a cyclic ring containing 3
to 10 carbon atoms.
[0199] The term "substituted" unless otherwise specified, refers to
substitution with any one or any combination of the following
substituents which may be the same or different and are
independently selected from hydrogen, hydroxy, halogen, carboxyl,
cyano, nitro, oxo (.dbd.O), thio (.dbd.S), substituted or
unsubstituted alkyl, substituted or unsubstituted alkoxy,
substituted or unsubstituted alkenyl, substituted or unsubstituted
alkynyl, substituted or unsubstituted aryl, substituted or
unsubstituted arylalkyl, substituted or unsubstituted cycloalkyl,
substituted or unsubstituted cycloalkylalkyl, substituted or
unsubstituted cycloalkenyl, substituted or unsubstituted
cycloalkenylalkyl, substituted or unsubstituted heteroaryl,
substituted or unsubstituted heteroarylalkyl, substituted or
unsubstituted heterocyclic ring, substituted heterocyclylalkyl
ring, substituted or unsubstituted guanidine, --COOR.sup.x,
--C(O)R.sup.x, --C(S)R.sup.x, --C(O)NR.sup.xR.sup.y,
--C(O)ONR.sup.xR.sup.y, --NR.sup.yR.sup.z,
--NR.sup.xCONR.sup.yR.sup.z, --N(R.sup.x)SOR.sup.y,
--N(R.sup.x)SO.sub.2R.sup.y, .dbd.N--N(R.sup.x)R.sup.y),
--NR.sup.xC(O)OR.sup.y, --NR.sup.xR.sup.y, --NR.sup.xC(O)R.sup.y--,
--NR.sup.xC(S)R.sup.y --NR.sup.xC(S)NR.sup.yR.sup.z,
--SONR.sup.xR.sup.y--, --SO.sub.2NR.sup.xR.sup.y--, --OR.sup.x,
--OR.sup.xC(O)NR.sup.yR.sup.z, --OR.sup.xC(O)OR.sup.y--,
--OC(O)R.sup.x, --OC(O)NR.sup.xR.sup.y,
--R.sup.xNR.sup.yC(O)R.sup.z, --R.sup.xOR.sup.y,
--R.sup.xC(O)OR.sup.y, --R.sup.xC(O)NR.sup.yR.sup.z,
--R.sup.xC(O)R.sup.x--, --R.sup.xOC(O)R.sup.y, --SR.sup.x--,
--SOR.sup.x, --SO.sub.2R.sup.x, and --ONO.sub.2, wherein R.sup.x,
R.sup.y and R.sup.z in each of the above groups can be hydrogen,
substituted or unsubstituted alkyl, substituted or unsubstituted
alkoxy, substituted or unsubstituted alkenyl, substituted or
unsubstituted alkynyl, substituted or unsubstituted aryl,
substituted or unsubstituted arylalkyl, substituted or
unsubstituted cycloalkyl, substituted or unsubstituted
cycloalkylalkyl, substituted or unsubstituted cycloalkenyl,
substituted or unsubstituted amino, substituted or unsubstituted
heteroaryl, substituted or unsubstituted heteroarylalkyl,
substituted or unsubstituted heterocyclic ring, or substituted
heterocyclylalkyl ring, or any two of R.sup.x, R.sup.y and R.sup.z
may be joined to form a substituted or unsubstituted saturated or
unsaturated 3-10 membered ring, which may optionally include
heteroatoms which may be the same or different and are selected
from O, NR.sup.x (e.g., R can be hydrogen or C.sub.1-6 alkyl) or S.
Substitution or the combinations of substituents envisioned by this
invention are preferably those that result in the formation of a
stable or chemically feasible compound. The term stable as used
herein refers to the compounds or the structure that are not
substantially altered when subjected to conditions to allow for
their production, detection and preferably their recovery,
purification and incorporation into a pharmaceutical composition.
The substituents in the aforementioned "substituted" groups cannot
be further substituted. For example, when the substituent on
"substituted alkyl" is "substituted aryl", the substituent on
"substituted aryl" cannot be "substituted alkenyl".
[0200] The term "halo", "halide", or, alternatively, "halogen"
means fluoro, chloro, bromo or iodo. The terms "haloalkyl,"
"haloalkenyl," "haloalkynyl" and "haloalkoxy" include alkyl,
alkenyl, alkynyl and alkoxy structures that are substituted with
one or more halo groups or with combinations thereof. For example,
the terms "fluoroalkyl" and "fluoroalkoxy" include haloalkyl and
haloalkoxy groups, respectively, in which the halo is fluorine.
[0201] The term "protecting group" or "PG" refers to a substituent
that is employed to block or protect a particular functionality.
Other functional groups on the compound may remain reactive. For
example, an "amino-protecting group" is a substituent attached to
an amino group that blocks or protects the amino functionality in
the compound. Suitable amino-protecting groups include, but are not
limited to, acetyl, trifluoroacetyl, tert-butoxycarbonyl (BOC),
benzyloxycarbonyl (CBz) and 9-fluorenylmethylenoxycarbonyl (Fmoc).
Similarly, a "hydroxy-protecting group" refers to a substituent of
a hydroxy group that blocks or protects the hydroxy functionality.
Suitable hydroxy-protecting groups include, but are not limited to,
acetyl and silyl. A "carboxy-protecting group" refers to a
substituent of the carboxy group that blocks or protects the
carboxy functionality. Suitable carboxy-protecting groups include,
but are not limited to, --CH.sub.2CH.sub.2SO.sub.2Ph, cyanoethyl,
2-(trimethylsilyl)ethyl, 2-(trimethylsilyl)ethoxymethyl,
2-(p-toluenesulfonyl)ethyl, 2-(p-nitrophenylsulfenyl)ethyl,
2-(diphenylphosphino)-ethyl, and nitroethyl. For a general
description of protecting groups and their use, see T. W. Greene,
Protective Groups in Organic Synthesis, John Wiley & Sons, New
York, 1991.
[0202] Certain of the compounds described herein contain one or
more asymmetric centers and can thus give rise to enantiomers,
diastereomers, and other stereoisomeric forms that can be defined,
in terms of absolute stereochemistry, as (R)- or (S)-. The present
chemical entities, pharmaceutical compositions and methods are
meant to include all such possible isomers, including racemic
mixtures, optically pure forms and intermediate mixtures.
Non-limiting examples of intermediate mixtures include a mixture of
isomers in a ratio of 10:90, 13:87, 17:83, 20:80, or 22:78.
Optically active (R)- and (S)-isomers can be prepared using chiral
synthons or chiral reagents, or resolved using conventional
techniques. When the compounds described herein contain olefinic
double bonds or other centers of geometric asymmetry, and unless
specified otherwise, it is intended that the compounds include both
E and Z geometric isomers.
[0203] The term "tautomers" refers to compounds, which are
characterized by relatively easy interconversion of isomeric forms
in equilibrium. These isomers are intended to be covered by this
invention. "Tautomers" are structurally distinct isomers that
interconvert by tautomerization. "Tautomerization" is a form of
isomerization and includes prototropic or proton-shift
tautomerization, which is considered a subset of acid-base
chemistry. "Prototropic tautomerization" or "proton-shift
tautomerization" involves the migration of a proton accompanied by
changes in bond order, often the interchange of a single bond with
an adjacent double bond. Where tautomerization is possible (e.g. in
solution), a chemical equilibrium of tautomers can be reached. An
example of tautomerization is keto-enol tautomerization. A specific
example of keto-enol tautomerization is the interconversion of
pentane-2,4-dione and 4-hydroxypent-3-en-2-one tautomers. Another
example of tautomerization is phenol-keto tautomerization. A
specific example of phenol-keto tautomerization is the
interconversion of pyridin-4-ol and pyridin-4(1H)-one
tautomers.
[0204] A "leaving group or atom" is any group or atom that will,
under the reaction conditions, cleave from the starting material,
thus promoting reaction at a specified site. Suitable examples of
such groups unless otherwise specified are halogen atoms and
mesyloxy, p-nitrobcnzensulphonyloxy and tosyloxy groups.
[0205] The term "prodrug" refers to a compound, which is an
inactive precursor of a compound, converted into its active form in
the body by normal metabolic processes. Prodrug design is discussed
generally in Hardma, et al. (Eds.), Goodman and Gilman's The
Pharmacological Basis of Therapeutics, 9th ed., pp. 11-16 (1996). A
thorough discussion is provided in Higuchi, et al., Prodrugs as
Novel Delivery Systems, Vol. 14, ASCD Symposium Series, and in
Roche (ed.), Bioreversible Carriers in Drug Design, American
Pharmaceutical Association and Pergamon Press (1987). To
illustrate, prodrugs can be converted into a pharmacologically
active form through hydrolysis of, for example, an ester or amide
linkage, thereby introducing or exposing a functional group on the
resultant product. The prodrugs can be designed to react with an
endogenous compound to form a water-soluble conjugate that further
enhances the pharmacological properties of the compound, for
example, increased circulatory half-life. Alternatively, prodrugs
can be designed to undergo covalent modification on a functional
group with, for example, glucuronic acid, sulfate, glutathione,
amino acids, or acetate. The resulting conjugate can be inactivated
and excreted in the urine, or rendered more potent than the parent
compound. High molecular weight conjugates also can be excreted
into the bile, subjected to enzymatic cleavage, and released back
into circulation, thereby effectively increasing the biological
half-life of the originally administered compound.
[0206] The term "ester" refers to a compound, which is formed by
reaction between an acid and an alcohol with elimination of water.
An ester can be represented by the general formula RCOOR'.
[0207] These prodrugs and esters are intended to be covered within
the scope of this invention.
[0208] Additionally the instant invention also includes the
compounds which differ only in the presence of one or more
isotopically enriched atoms for example replacement of hydrogen
with deuterium or tritium, or the replacement of a carbon by
.sup.13C- or .sup.14C-enriched carbon.
[0209] The compounds of the present invention may also contain
unnatural proportions of atomic isotopes at one or more of atoms
that constitute such compounds. For example, the compounds may be
radiolabeled with radioactive isotopes, such as for example tritium
(.sup.3H), iodine-125 (.sup.125I) or carbon-14 (.sup.14C). All
isotopic variations of the compounds of the present invention,
whether radioactive or not, are encompassed within the scope of the
present invention.
[0210] Pharmaceutically acceptable salts forming part of this
invention include salts derived from inorganic bases such as Li,
Na, K, Ca, Mg, Fe, Cu, Zn, and Mn; salts of organic bases such as
N,N'-diacctylcthylencdiamine, glucamine, tricthylamine, choline,
hydroxide, dicyclohexylamine, metformin, benzylamine,
trialkylamine, and thiamine; chiral bases such as alkylphenylamine,
glycinol, and phenyl glycinol; salts of natural amino acids such as
glycine, alanine, valine, leucine, isoleucine, norleucine,
tyrosine, cystine, cysteine, methionine, proline, hydroxy proline,
histidine, omithine, lysine, arginine, and serine; quaternary
ammonium salts of the compounds of invention with alkyl halides,
alkyl sulphates such as MeI and (Me).sub.2SO.sub.4; non-natural
amino acids such as D-isomers or substituted amino acids;
guanidine; and substituted guanidine wherein the substituents are
selected from nitro, amino, alkyl, alkenyl, alkynyl, ammonium or
substituted ammonium salts and aluminum salts. Salts may include
acid addition salts where appropriate which are sulphates,
nitrates, phosphates, perchlorates, borates, hydrohalides (e.g.,
hydrochlorides), acetates, tartrates, maleates, citrates,
fumarates, succinates, palmoates, methanesulphonates, benzoates,
salicylates, benzenesulfonates, ascorbates, glycerophosphates, and
ketoglutarates.
[0211] When ranges are used herein for physical properties, such as
molecular weight, or chemical properties, such as chemical
formulae, all combinations and subcombinations of ranges and
specific embodiments therein are intended to be included. The term
"about" when referring to a number or a numerical range means that
the number or numerical range referred to is an approximation
within experimental variability (or within statistical experimental
error), and thus the number or numerical range may vary from, for
example, between 1% and 15% of the stated number or numerical
range. The term "comprising" (and related terms such as "comprise"
or "comprises" or "having" or "including") includes those
embodiments, for example, an embodiment of any composition of
matter, composition, method, or process, or the like, that "consist
of" or "consist essentially of" the described features.
[0212] The following abbreviations and terms have the indicated
meanings throughout: AIDS=Acquired Immuno Deficiency Syndrome;
HIV=Human Innunodeficiency Virus; Abbreviations used herein have
their conventional meaning within the chemical and biological
arts.
[0213] The term "cell proliferation" refers to a phenomenon by
which the cell number has changed as a result of division. This
term also encompasses cell growth by which the cell morphology has
changed (e.g., increased in size) consistent with a proliferative
signal.
[0214] The term "co-administration," "administered in combination
with," and their grammatical equivalents, as used herein,
encompasses administration of two or more agents to an animal so
that both agents and/or their metabolites are present in the animal
at the same time. Co-administration includes simultaneous
administration in separate compositions, administration at
different times in separate compositions, or administration in a
composition in which both agents are present.
[0215] The term "effective amount" or "therapeutically effective
amount" refers to that amount of a compound described herein that
is sufficient to effect the intended application including but not
limited to disease treatment, as defined below. The therapeutically
effective amount may vary depending upon the intended application
(in vitro or in vivo), or the subject and disease condition being
treated, e.g., the weight and age of the subject, the severity of
the disease condition, the manner of administration and the like,
which can readily be determined by one of ordinary skill in the
art. The term also applies to a dose that will induce a particular
response in target cells, e.g. reduction of platelet adhesion
and/or cell migration. The specific dose will vary depending on the
particular compounds chosen, the dosing regimen to be followed,
whether it is administered in combination with other compounds,
timing of administration, the tissue to which it is administered,
and the physical delivery system in which it is carried. In one
embodiment, the amount of compound administered ranges from about
0.1 mg to 5 g, from about 1 mg to 2.0 g, from about 100 mg to 1.5
g, from about 200 mg to 1.5 g, from about 400 mg to 1.5 g, and from
about 400 mg to 1.0 g.
[0216] As used herein, "treatment," "treating," or "ameliorating"
are used interchangeably. These terms refers to an approach for
obtaining beneficial or desired results including but not limited
to therapeutic benefit and/or a prophylactic benefit. By
therapeutic benefit is meant eradication or amelioration of the
underlying disorder being treated. Also, a therapeutic benefit is
achieved with the eradication or amelioration of one or more of the
physiological symptoms associated with the underlying disorder such
that an improvement is observed in the patient, notwithstanding
that the patient may still be afflicted with the underlying
disorder. For prophylactic benefit, the compositions may be
administered to a patient at risk of developing a particular
disease, or to a patient reporting one or more of the physiological
symptoms of a disease, even though a diagnosis of this disease may
not have been made.
[0217] A "therapeutic effect," as that term is used herein,
encompasses a therapeutic benefit and/or a prophylactic benefit as
described above. A prophylactic effect includes delaying or
eliminating the appearance of a disease or condition, delaying or
eliminating the onset of symptoms of a disease or condition,
slowing, halting, or reversing the progression of a disease or
condition, or any combination thereof.
[0218] The term "subject" or "patient" refers to an animal, such as
a mammal, for example a human. The methods described herein can be
useful in both human therapeutics and veterinary applications
(e.g., dogs, cats, cows, sheep, pigs, horses, goats, chickens,
turkeys, ducks, and geese).
[0219] In some embodiments, the patient is a mammal, and in some
embodiments, the patient is human.
[0220] "Radiation therapy" means exposing a patient, using routine
methods and compositions known to the practitioner, to radiation
emitters such as alpha-particle emitting radionuclides (e.g.,
actinium and thorium radionuclides), low linear energy transfer
(LET) radiation emitters (i.e. beta emitters), conversion electron
emitters (e.g. strontium-89 and samarium-153-EDTMP), or high-energy
radiation, including without limitation x-rays, gamma rays, and
neutrons.
[0221] The term "pharmaceutically acceptable excipient" includes,
but is not limited to, any and all solvents, dispersion media,
coatings, antibacterial and antifungal agents, isotonic and
absorption delaying agents, one or more suitable diluents, fillers,
salts, disintegrants, binders, lubricants, glidants, wetting
agents, controlled release matrices, colorants/flavoring, carriers,
buffers, stabilizers, solubilizers, and combinations thereof.
Except insofar as any conventional media or agent is incompatible
with the active ingredient, its use in the therapeutic compositions
of the invention is contemplated. Supplementary active ingredients
can also be incorporated into the compositions.
[0222] "Inflammatory response" as used herein is characterized by
redness, heat, swelling and pain (i.e., inflammation) and typically
involves tissue injury or destruction. An inflammatory response is
usually a localized, protective response elicited by injury or
destruction of tissues, which serves to destroy, dilute or wall off
(sequester) both the injurious agent and the injured tissue.
Inflammatory responses are notably associated with the influx of
leukocytes and/or leukocyte (e.g., neutrophil) chemotaxis.
Inflammatory responses may result from infection with pathogenic
organisms and viruses, noninfectious means such as trauma or
reperfusion following myocardial infarction or stroke, immune
responses to foreign antigens, and autoimmune diseases.
Inflammatory responses amenable to treatment with the methods and
compounds according to the invention encompass conditions
associated with reactions of the specific defence system as well as
conditions associated with reactions of the non-specific defence
system.
[0223] The therapeutic methods of the invention include methods for
the amelioration of conditions associated with inflammatory cell
activation. "Inflammatory cell activation" refers to the induction
by a stimulus (including but not limited to, cytokines, antigens or
auto-antibodies) of a proliferative cellular response, the
production of soluble mediators (including but not limited to
cytokines, oxygen radicals, enzymes, prostanoids, or vasoactive
amines), or cell surface expression of new or increased numbers of
mediators (including but not limited to, major histocompatibility
antigens or cell adhesion molecules) in inflammatory cells
(including but not limited to monocytes, macrophages, T
lymphocytes, B lymphocytes, granulocytes (polymorphonuclear
leukocytes including neutrophils, basophils, and eosinophils) mast
cells, dendritic cells, Langerhans cells, and endothelial cells).
It will be appreciated by persons skilled in the art that the
activation of one or a combination of these phenotypes in these
cells can contribute to the initiation, perpetuation, or
exacerbation of an inflammatory condition.
[0224] "Autoimmune disease" as used herein refers to any group of
disorders in which tissue injury is associated with humoral or
cell-mediated responses to the body's own constituents. "Transplant
rejection" (or "transplantation rejection") as used herein refers
to any immune response directed against grafted tissue (including
organs or cells (e.g., bone marrow), characterized by a loss of
function of the grafted and surrounding tissues, pain, swelling,
leukocytosis, and thrombocytopenia). "Allergic disease" as used
herein refers to any symptoms, tissue damage, or loss of tissue
function resulting from allergy. "Arthritic disease" as used herein
refers to any disease that is characterized by inflammatory lesions
of the joints attributable to a variety of etiologies. "Dermatitis"
as used herein refers to any of a large family of diseases of the
skin that are characterized by inflammation of the skin
attributable to a variety of etiologies.
[0225] The methods of the invention may be applied to cell
populations in vivo or ex vivo. "In vivo" means within a living
individual, as within an animal or human or in a subject's body. In
this context, the methods of the invention may be used
therapeutically or prophylactically in an individual. "Ex vivo" or
"In vitro" means outside of a living individual. Examples of ex
vivo cell populations include in vitro cell cultures and biological
samples including but not limited to fluid or tissue samples
obtained from individuals. Such samples may be obtained by methods
known in the art. Exemplary biological fluid samples include blood,
cerebrospinal fluid, urine, and saliva. Exemplary tissue samples
include tumors and biopsies thereof. In this context, the invention
may be used for a variety of purposes, including therapeutic and
experimental purposes. For example, the invention may be used ex
vivo or in vitro to determine the optimal schedule and/or dosing of
administration of a glutaminase inhibitor for a given indication,
cell type, individual, and other parameters. Information gleaned
from such use may be used for experimental or diagnostic purposes
or in the clinic to set protocols for in vivo treatment. Other ex
vivo uses for which the invention may be suited are described below
or will become apparent to those skilled in the art.
Pharmaceutical Compositions
[0226] The invention provides a pharmaceutical composition
comprising one or more compounds of the present invention. The
pharmaceutical composition may include one or more additional
active ingredients as described herein. The pharmaceutical
composition may be administered for any of the disorders described
herein.
[0227] The subject pharmaceutical compositions are typically
formulated to provide a therapeutically effective amount of a
compound of the present invention as the active ingredient. Where
desired, the pharmaceutical compositions contain a compound of the
present invention as the active ingredient and one or more
pharmaceutically acceptable carriers or excipients, such as inert
solid diluents and fillers, diluents, including sterile aqueous
solution and various organic solvents, permeation enhancers,
solubilizers and adjuvants.
[0228] The pharmaceutical compositions can be administered alone or
in combination with one or more other agents, which are also
typically administered in the form of pharmaceutical compositions.
Where desired, the subject compounds and other agent(s) may be
mixed into a preparation or both components may be formulated into
separate preparations to use them in combination separately or at
the same time.
[0229] Methods include administration of a compound of the present
invention by itself, or in combination as described herein, and in
each case optionally including one or more suitable diluents,
fillers, salts, disintegrants, binders, lubricants, glidants,
wetting agents, controlled release matrices, colorants/flavoring,
carriers, excipients, buffers, stabilizers, solubilizers, and
combinations thereof.
[0230] Preparations of various pharmaceutical compositions are
known in the art. See, e.g., Anderson, Philip O.; Knoben, James E.;
Troutman, William G, eds., Handbook of Clinical Drug Data, Tenth
Edition, McGraw-Hill, 2002; Pratt and Taylor, eds., Principles of
Drug Action, Third Edition, Churchill Livingston, N.Y., 1990;
Katzung, ed., Basic and Clinical Pharmacology, Ninth Edition,
McGraw Hill, 2003; Goodman and Gilman, eds., The Pharmacological
Basis of Therapeutics, Tenth Edition, McGraw Hill, 2001; Remingtons
Pharmaceutical Sciences, 20th Ed., Lippincott Williams &
Wilkins., 2000; Martindale, The Extra Pharmacopoeia, Thirty-Second
Edition (The Pharmaceutical Press, London, 1999), all of which are
incorporated by reference herein in their entirety.
[0231] The compounds or pharmaceutical composition of the present
invention can be administered by any route that enables delivery of
the compounds to the site of action, such as oral routes,
intraduodenal routes, parenteral injection (including intravenous,
intraarterial, subcutaneous, intramuscular, intravascular,
intraperitoneal or infusion), topical administration (e.g.
transdermal application), rectal administration, via local delivery
by catheter or stent or through inhalation. The compounds can also
be administered intraadiposally or intrathecally.
[0232] The compositions can be administered in solid, semi-solid,
liquid or gaseous form, or may be in dried powder, such as
lyophilized form. The pharmaceutical compositions can be packaged
in forms convenient for delivery, including, for example, solid
dosage forms such as capsules, sachets, cachets, gelatins, papers,
tablets, capsules, suppositories, pellets, pills, troches, and
lozenges. The type of packaging will generally depend on the
desired route of administration. Implantable sustained release
formulations are also contemplated, as are transdermal
formulations.
Method of Treatment
[0233] The invention also provides methods of using the compounds
or pharmaceutical compositions of the present invention to treat
disease conditions, including, but not limited to, diseases
associated with overexpression of glutaminase and/or due to an
excess of glutamine.
[0234] The treatment methods provided herein comprise administering
to the subject a therapeutically effective amount of a compound of
the invention. In one embodiment, the present invention provides a
method of treating an inflammation disorder, including autoimmune
diseases in a mammal. The method comprises administering to the
mammal a therapeutically effective amount of a compound of the
present invention.
[0235] It will be appreciated that the treatment methods of the
invention are useful in the fields of human medicine and veterinary
medicine. Thus, the individual to be treated may be a mammal,
preferably human, or other animal. For veterinary purposes,
individuals include but are not limited to farm animals including
cows, sheep, pigs, horses, and goats; companion animals such as
dogs and cats; exotic and/or zoo animals; laboratory animals
including mice, rats, rabbits, guinea pigs, and hamsters; and
poultry such as chickens, turkeys, ducks, and geese.
[0236] In some embodiments, the method of treating inflammatory or
autoimmune diseases comprises administering to a subject (e.g. a
mammal) a therapeutically effective amount of one or more compounds
of the present invention that inhibits glutaminase. Such inhibition
of glutaminase may be advantageous for treating any of the diseases
or conditions described herein. For example, inhibition of
glutaminase may inhibit inflammatory responses associated with
inflammatory diseases, autoinmmune disease, or diseases related to
an undesirable immune response including but not limited to asthma,
emphysema, allergy, dermatitis, rheumatoid arthritis, psoriasis,
lupus erythematosus, or graft versus host disease. Inhibition of
glutaminase may further provide for a reduction in the inflammatory
or undesirable immune response without a concomittant reduction in
the ability to reduce a bacterial, viral, and/or fungal
infection.
[0237] In other embodiments, the present invention provides methods
of using the compounds or pharmaceutical compositions to treat
respiratory diseases including but not limited to diseases
affecting the lobes of lung, pleural cavity, bronchial tubes,
trachea, upper respiratory tract, or the nerves and muscle for
breathing. For example, methods are provided to treat obstructive
pulmonary disease. Chronic obstructive pulmonary disease (COPD) is
an umbrella term for a group of respiratory tract diseases that are
characterized by airflow obstruction or limitation. Conditions
included in this umbrella term include chronic bronchitis,
emphysema, and bronchiectasis.
[0238] In another embodiment, the compounds described herein are
used for the treatment of asthma. Also, the compounds or
pharmaceutical compositions described herein may be used for the
treatment of endotoxemia and sepsis. In one embodiment, the
compounds or pharmaceutical compositions described herein are used
to for the treatment of rheumatoid arthritis (RA). In yet another
embodiment, the compounds or pharmaceutical compositions described
herein is used for the treatment of contact or atopic dermatitis.
Contact dermatitis includes irritant dermatitis, phototoxic
dermatitis, allergic dermatitis, photoallergic dermatitis, contact
urticaria, systemic contact-type dermatitis and the like. Irritant
dermatitis can occur when too much of a substance is used on the
skin of when the skin is sensitive to certain substance. Atopic
dermatitis, sometimes called eczema, is a kind of dermatitis, an
atopic skin disease.
[0239] The invention also relates to a method of treating a
hyperproliferative disorder in a mammal that comprises
administering to said mammal a therapeutically effective amount of
a compound of the present invention, or a pharmaceutically
acceptable salt, ester, prodrug, solvate, hydrate or derivative
thereof. In some embodiments, said method relates to the treatment
of cancer such as acute myeloid leukemia, thymus, brain, lung,
squamous cell, skin, eye, retinoblastoma, intraocular melanoma,
oral cavity and oropharyngeal, bladder, gastric, stomach,
pancreatic, bladder, breast, cervical, head, neck, renal, kidney,
liver, ovarian, prostate, colorectal, esophageal, testicular,
gynecological, thyroid, CNS, PNS, AIDS-related (e.g. lymphoma and
Kaposi's sarcoma) or viral-induced cancer. In some embodiments,
said method relates to the treatment of a non-cancerous
hyperproliferative disorder such as benign hyperplasia of the skin
(e. g., psoriasis), restenosis, or prostate (e.g., benign prostatic
hypertrophy (BPH)).
[0240] The invention also relates to a method of treating diseases
related to vasculogenesis or angiogenesis in a mammal that
comprises administering to said mammal a therapeutically effective
amount of a compound of the present invention. In some embodiments,
said method is for treating a disease selected from the group
consisting of tumor angiogenesis, chronic inflammatory disease such
as rheumatoid arthritis, atherosclerosis, inflammatory bowel
disease, skin diseases such as psoriasis, eczema, and scleroderma,
diabetes, diabetic retinopathy, retinopathy of prematurity,
age-related macular degeneration, hemangioma, glioma, melanoma,
Kaposi's sarcoma and ovarian, breast, lung, pancreatic, prostate,
colon and epidermoid cancer.
[0241] Patients that can be treated with compounds of the present
invention according to the methods of this invention include, for
example, patients that have been diagnosed as having psoriasis;
restenosis; atherosclerosis; BPH; breast cancer such as a ductal
carcinoma in duct tissue in a mammary gland, medullary carcinomas,
colloid carcinomas, tubular carcinomas, and inflammatory breast
cancer; ovarian cancer, including epithelial ovarian tumors such as
adcnocarcinoma in the ovary and an adenocarcinoma that has migrated
from the ovary into the abdominal cavity; uterine cancer; cervical
cancer such as adenocarcinoma in the cervix epithelial including
squamous cell carcinoma and adenocarcinomas; prostate cancer, such
as a prostate cancer selected from the following: an adenocarcinoma
or an adenocarinoma that has migrated to the bone; pancreatic
cancer such as epitheliod carcinoma in the pancreatic duct tissue
and an adenocarcinoma in a pancreatic duct; bladder cancer such as
a transitional cell carcinoma in urinary bladder, urothelial
carcinomas (transitional cell carcinomas), tumors in the urothelial
cells that line the bladder, squamous cell carcinomas,
adenocarcinomas, and small cell cancers; leukemia such as acute
mycloid leukemia (AML), acute lymphocytic leukemia, chronic
lymphocytic leukemia, chronic myeloid leukemia, hairy cell
leukemia, myelodysplasia, myeloproliferative disorders, acute
myelogenous leukemia (AML), chronic myelogenous leukemia (CML),
mastocytosis, chronic lymphocytic leukemia (CLL), multiple myeloma
(MM), and myelodysplastic syndrome (MDS); bone cancer; lung cancer
such as non-small cell lung cancer (NSCLC), which is divided into
squamous cell carcinomas, adenocarcinomas, and large cell
undifferentiated carcinomas, and small cell lung cancer; skin
cancer such as basal cell carcinoma, melanoma, squamous cell
carcinoma and actinic keratosis, which is a skin condition that
sometimes develops into squamous cell carcinoma; eye
retinoblastoma; cutaneous or intraocular (eye) melanoma; primary
liver cancer (cancer that begins in the liver); kidney cancer;
thyroid cancer such as papillary, follicular, medullary and
anaplastic; AIDS-related lymphoma such as diffuse large B-cell
lymphoma, B-cell immunoblastic lymphoma and small non-clcaved cell
lymphoma; Kaposi's Sarcoma; viral-induced cancers including
hepatitis B virus (HBV), hepatitis C virus (HCV), and
hepatocellular carcinoma; human lymphotropic virus-type 1 (HTLV-I)
and adult T-cell leukemia/lymphoma; and human papilloma virus (HPV)
and cervical cancer; central nervous system cancers (CNS) such as
primary brain tumor, which includes gliomas (astrocytoma,
anaplastic astrocytoma, or glioblastoma multiforme),
Oligodendroglioma, Ependymoma, Meningioma, Lymphoma, Schwannoma,
and Medulloblastoma; peripheral nervous system (PNS) cancers such
as acoustic neuromas and malignant peripheral nerve sheath tumor
(MPNST) including neurofibromas and schwannomas, malignant fibrous
cytoma, malignant fibrous histiocytoma, malignant meningioma,
malignant mesothelioma, and malignant mixed Mtllerian tumor; oral
cavity and oropharyngeal cancer such as, hypopharyngeal cancer,
laryngeal cancer, nasopharyngeal cancer, and oropharyngeal cancer;
stomach cancer such as lymphomas, gastric stromal tumors, and
carcinoid tumors; testicular cancer such as germ cell tumors
(GCTs), which include seminomas and nonseminomas, and gonadal
stromal tumors, which include Leydig cell tumors and Sertoli cell
tumors; thymus cancer such as to thymomas, thymic carcinomas,
Hodgkin disease, non-Hodgkin lymphomas carcinoids or carcinoid
tumors; rectalcancer; and colon cancer.
[0242] In another aspect, the present invention provides methods of
disrupting the function of a leukocyte or disrupting a function of
an osteoclast. The method includes contacting the leukocyte or the
osteoclast with a function disrupting amount of a compound of the
invention.
[0243] In another aspect of the present invention, methods are
provided for treating ophthalmic disease by administering one or
more compounds or pharmaceutical compositions described herein to
the eye of a subject.
[0244] The invention further provides methods of inhibiting
glutaminase by contacting a glutaminase with an amount of a
compound of the invention sufficient to inhibit the activity of the
glutaminase enzyme. In some embodiments, the invention provides
methods of inhibiting glutaminase enzyme activity by contacting a
glutaminase enzyme with an amount of a compound of the invention
sufficient to inhibit the activity of the glutaminase enzyme. In
some embodiments, the invention provides methods of inhibiting
glutaminase enzyme activity. Such inhibition can take place in
solution, in a cell expressing one or more glutaminase enzyme, in a
tissue comprising a cell expressing the glutaminase, or in an
organism expressing glutaminase. In some embodiments, the invention
provides methods of inhibiting glutaminase activity in an animal
(including mammal such as humans) by contacting said animal with an
amount of a compound of the invention sufficient to inhibit the
activity of the glutaminase enzyme in said animal.
[0245] The following general methodology described herein provides
the manner and process of making and using the compounds of the
present invention and are illustrative rather than limiting.
Further modification of provided methodology and additionally new
methods may also be devised in order to achieve and serve the
purpose of the invention. Accordingly, it should be understood that
there may be other embodiments which fall within the spirit and
scope of the invention as defined by the specification hereto.
[0246] Illustrative compounds of the present invention include
those specified above in Table 1 and pharmaceutically acceptable
salts thereof. The present invention should not be construed to be
limited to only these compounds.
General Methods of Preparation
[0247] The compounds of the present invention may be prepared by
the following processes. Unless otherwise indicated, the variables
(e.g. R.sup.1, R.sup.2, P, Q, A, B and L) when used in the below
formulae are to be understood to represent those groups described
above in relation to formula (I). These methods can similarly be
applied to other compounds of formula (I) as provided herein above
with or without modification.
[0248] Scheme 1: This scheme provides a method for the preparation
of compounds of formula (I) wherein R.sup.1 and R.sup.2 are
independently substituted or unsubstituted aryl or substituted or
unsubstituted heteroaryl, P and Q are independently
--NR.sup.xC(O)--(CR.sup.xR.sup.y).sub.r-- or
--C(R.sup.xR.sup.y).sub.r--C(O)--NR.sup.x--, L is
-L.sub.1-L.sub.2-L.sub.3-, wherein L.sub.2 is substituted or
unsubstituted 3 to 14 membered heterocyclyl, L.sub.1 and L.sub.3
are absent or substituted or unsubstituted C.sub.1-6 alkyl (such as
methyl), A is
##STR00064##
##STR00065##
r is 0 or 1, and all the other variables (including R.sup.x and
R.sup.y) are as described above in relation to formula (I).
##STR00066##
A compound of formula (1) (wherein s and v are 0 or 1 and R is
alkyl) can be coupled with a compound of formula (2) to form a
compound of formula (3). The compound of formula (3) can be reduced
to form a compound of formula (4) using a suitable reducing agent
such as, for example, Fe/NH.sub.4Cl, EtOH and water. A compound of
formula (4) can then be coupled with a compound of formula (5) to
form a compound of formula (6), e.g., in the presence of HATU, DMF
and DIPEA. The compound of formula (6) can then be hydrolysed to
give a compound of formula (7), which can then be reacted with a
compound of formula (8) in the presence of POCl.sub.3 to form a
compound of formula (9), The compound of formula (9) can then be
coupled with a compound of formula (10), e.g., in the presence of
HATU, DMF and DIPEA to form the compound of formula (I). This
scheme is illustrated in Illustrations 1 and 2 below.
##STR00067## ##STR00068##
[0249] Scheme 2: This scheme provides a method for the preparation
of compounds of formula (I) wherein R.sup.1 and R.sup.2 are
independently substituted or unsubstituted aryl or substituted or
unsubstituted heteroaryl, P and Q are independently
--NR.sup.xC(O)--(CR.sup.xR.sup.y).sub.r-- or
--C(R.sup.xR.sup.y).sub.r--C(O)--NR.sup.x--, L is
-L.sub.1-L.sub.2-L.sub.3-, wherein L.sub.2 is substituted or
unsubstituted 3 to 14 membered heterocyclyl, L.sub.1 and L.sub.3
are absent or substituted or unsubstituted C.sub.1-6 alkyl (such as
methyl), A is
##STR00069##
B is
##STR00070##
[0250] r is 0 or 1, and all the other variables (including R.sup.x
and R.sup.y) are as described above in relation to formula (I).
##STR00071##
A compound of formula (11) can be coupled with a compound of
formula (12) wherein X is a leaving group to form a compound of
formula (13). The compound of formula (13) can be reacted with a
compound of formula (14) to form a compound of formula (15), which
can then be reacted with a compound of formula (8), for example, in
the presence of POCl.sub.3, to form a compound of formula (16). The
compound of formula (16) can be coupled with a compound of formula
R.sup.2--CH.sub.2--COOH to afford the compound of formula (I),
wherein the variables are as defined above.
##STR00072##
A compound of formula (12) can be coupled with a compound of
formula (14a) to form a compound of formula (17). The compound of
formula (17) can be reacted with a compound of formula
R.sup.1--CH.sub.2--COOH followed by ester hydrolysis to form a
compound of formula (15). The compound of formula (15) can then be
reacted with a compound of formula (8), for example, in the
presence of POCl.sub.3, to form a compound of formula (16), which
can be coupled with a compound of formula R.sup.2--CH.sub.2--COOH
to give a compound of formula (I), wherein the variables are as
defined above. This scheme is illustrated in Illustrations 1 and 2
below.
##STR00073## ##STR00074##
[0251] Scheme 3: This scheme provides a method for the preparation
of a compound of formula (I) wherein R.sup.1 and R.sup.2 are
independently substituted or unsubstituted aryl or substituted or
unsubstituted heteroaryl, P and Q are independently
--NR.sup.xC(O)--(CR.sup.xC.sup.y).sub.r-- or
C(R.sup.xR.sup.y).sub.r--C(O)--NR.sup.x--, L is
-L.sub.1-L.sub.2-L.sub.3-, wherein L.sub.2 is substituted or
unsubstituted 3 to 14 membered heterocyclyl, L.sub.1 and L.sub.3
are absent or substituted or unsubstituted C.sub.1-6 alkyl (such as
methyl), A is
##STR00075##
B is
##STR00076##
[0252] r is 0 or 1, s is 0 or 1, v is or 1 and all the other
variables (including R.sup.x and R.sup.y) are as described above in
relation to formula (I).
Step-1
##STR00077##
[0253] Step-2
##STR00078##
[0254] Step-1: The compound of formula (18) wherein FG is nitro or
amino and X is a leaving group such as bromine can be coupled with
a compound of formula (19) wherein Pg is a protecting group
optionally in the presence of tetrabutylammonium iodide (TBAI) and
a suitable base such as K.sub.2CO.sub.3 to form a compound of
formula (20). The compound of formula (20) (wherein FG.sub.1 is
nitro (--NO.sub.2)) can be reduced to form a compound of formula
(21), which can be coupled with a compound of formula (22) to form
the compound of formula (23). Alternatively, the compound of
formula (20) (wherein FG.sub.2 is amino (--NH.sub.2)) can be
coupled with a compound of formula (22) to form compound of formula
(23). Step-2: The compound of formula (23) can be deprotected to
form a compound of formula (23a). The compound of formula (23a) can
be reacted with a compound of formula (8), for example in the
presence of POCl.sub.3, to form a compound of formula (24), which
can be coupled with a compound of formula (25) in the presence of
suitable reagents such as HATU and DIPEA to form the compound of
formula (I). This scheme is illustrated in Illustrations 1 and 2
below.
Illustration 1
Step-1:
##STR00079##
[0255] Step-2:
##STR00080##
[0256] Illustration 2
Step-1:
##STR00081##
[0257] Step-2:
##STR00082##
[0259] Scheme 4: This scheme provides a method for the preparation
of a compound of formula (I) wherein R.sup.1 and R.sup.2 are
independently substituted or unsubstituted aryl or substituted or
unsubstituted heteroaryl, P and Q are independently
--NR.sup.xC(O)--(CR.sup.xR.sup.y).sub.r-- or
--C(R.sup.xR.sup.y).sub.r--C(O)--NR.sup.x--, L is
-L.sub.1-L.sub.2-L.sub.3-wherein L.sub.2 is substituted or
unsubstituted 3 to 14 membered heterocyclyl, L.sub.1 and L.sub.3
are absent or substituted or unsubstituted C.sub.1-6 alkyl (such as
methyl), A is
##STR00083##
B is
##STR00084##
[0260] r is 0 or 1, s is 0 or 1, v is 0 or 1 and all other
variables (including R and R.sup.y) are as described above in
relation to formula (I).
Step-1:
##STR00085##
[0261] Step-2:
##STR00086##
[0262] Step-1: The compound of formula (18) wherein FG is nitro or
amino and X is a leaving group such as bromine can be coupled with
a compound of formula (19) wherein Pg is a protecting group, for
example in the presence of tetrabutylammonium iodide (TBAI) and a
suitable base such as K.sub.2CO.sub.3 to form a compound of formula
(20). The compound of formula (20) can be converted using aqueous
ammonia to form a compound of formula (26), which can be converted
using, for example thionyl chloride, to form a compound of formula
(27). The compound of formula (27) (wherein FG.sub.1 is nitro
(--NO.sub.2)) can be reduced to form compound (28), which can be
coupled with a compound of formula (22) in the presence of suitable
reagents such as HATU and DIPEA to form a compound of formula (29).
Alternatively, the compound of formula (27) (wherein FG.sub.2 is
amino (--NH.sub.2)) can be coupled with a compound of formula (22)
in the presence of suitable reagents such as HATU and DIPEA to form
a compound of formula (29). Step-2: The compound of formula (29)
can be reacted with a compound of formula (8) to form a compound of
formula (24). The compound of formula (24) can be coupled with a
compound of formula (25) in the presence of suitable reagents such
as HATU and DIPEA to form the compound of formula (I). This scheme
is illustrated in Illustrations 1 and 2 below.
Illustration
Step-1:
##STR00087##
[0263] Step-2:
##STR00088##
[0265] Scheme 5: This scheme provides a method for the preparation
of a compound of formula (I) wherein R.sup.1 and R.sup.2 are
independently substituted or unsubstituted aryl or substituted or
unsubstituted heteroaryl, P and Q are independently
--NR.sup.xC(O)--(CR.sup.xR.sup.y).sub.r-- or
--C(R.sup.xR.sup.y).sub.r--C(O)--NR.sup.x--, L is
-L.sub.1-L.sub.2-L.sub.3-wherein L.sub.2 is substituted or
unsubstituted 3 to 14 membered heterocyclyl, L.sub.1 and L.sub.3
are absent or substituted or unsubstituted C.sub.1-6 alkyl (such as
methyl), A is
##STR00089##
B is
##STR00090##
[0266] r is 0 or 1, s is 0 or 1, v is 0 or 1 and all other
variables (including R.sup.x and R.sup.y) are as described above in
relation to formula (I).
Step-1
##STR00091##
[0267] Step-2
##STR00092##
[0268] Step-1: The compound of formula (18) wherein FG is nitro or
amino and X is a leaving group such as bromine can be coupled with
a compound of formula (30) to form a compound of formula (27). The
compound of formula (27) (wherein FG.sub.1 is nitro (--NO.sub.2))
can be reduced to form compound (28), which can be coupled with a
compound of formula (22) in the presence of suitable reagents such
as HATU and DIPEA to form a compound of formula (29).
Alternatively, a compound of formula (27) (wherein FG.sub.2 is
amino (--NH.sub.2)) can be coupled with a compound of formula (22)
in the presence of suitable reagents such as HATU and DIPEA to form
a compound of formula (29). Step-2: The compound of formula (29)
can be reacted with a compound of formula (8) to form a compound of
formula (24), which can be coupled with a compound of formula (25)
in the presence of suitable reagents such as HATU and DIPEA to form
the compound of formula (I). This scheme is illustrated in
Illustrations 1 and 2 below.
Illustration
Step-1
##STR00093##
[0269] Step-2:
##STR00094##
[0271] Similar methodologies with modifications known to those
skilled in the art can be used to synthesize compounds of formula
of (I), (II) and (III) wherein all the variables are to be
understood to represent those groups described above using suitable
intermediates and reagents.
Experimental
[0272] The examples and preparations provided below further
illustrate and exemplify the compounds of the present invention and
methods of preparing such compounds. It is to be understood that
the scope of the present invention is not limited in any way by the
scope of the following examples and preparations. In the following
examples, molecules with a single chiral center, unless otherwise
noted, exist as a racemic mixture. Those molecules with two or more
chiral centers, unless otherwise noted, exist as a racemic mixture
of diastereomers. Single enantiomners/diastereomers may be obtained
by methods known to those skilled in the art.
TABLE-US-00003 TABLE OF INTERMEDIATES: No. Structure 1 ##STR00095##
2 ##STR00096## 3 ##STR00097## 4 ##STR00098## 5 ##STR00099## 6
##STR00100## 7 ##STR00101## 8 ##STR00102## 9 ##STR00103## 10
##STR00104## 11 ##STR00105## 12 ##STR00106## 13 ##STR00107## 14
##STR00108## 15 ##STR00109## 16 ##STR00110## 17 ##STR00111## 18
##STR00112## 19 ##STR00113## 20 ##STR00114## 21 ##STR00115## 22
##STR00116## 23 ##STR00117## 24 ##STR00118## 25 ##STR00119## 26
##STR00120## 27 ##STR00121## 28 ##STR00122## 29 ##STR00123## 30
##STR00124## 31 ##STR00125## 32 ##STR00126## 33 ##STR00127## 34
##STR00128## 35 ##STR00129## 36 ##STR00130## 37 ##STR00131## 38
##STR00132## 39 ##STR00133## 40 ##STR00134## 41 ##STR00135##
[0273] Intermediate 1: Ethyl
1-(6-nitropyridin-3-yl)piperidine-4-carboxylate:
3-Bromo-6-nitropyridine (2 g, 9.85 mmol), Ethyl isonipecotate (1.7
g, 10.8 mmol), K.sub.2CO.sub.3 (1.36 g, 9.84 mmol) and
tetrabutylammonium iodide were taken in DMSO (10 ml). This mixture
was stirred at 100.degree. C. under inert atmosphere for 16 hrs.
After completion of the reaction, reaction was cooled to rt and
diluted with water. Aqueous layer was extracted with EtOAc. Organic
layer was dried on anhydrous Na.sub.2SO.sub.4. EtOAc was removed on
rotavapour to obtain crude. Crude was purified on combi-flash using
EtOAc and Petether (1:2) as eluent to afford the titled compound
(2.2 g) as a yellow solid. .sup.1H-NMR (.delta. ppm, DMSO-d.sub.6,
400 MHz): 8.16-8.10 (m, 2H), 7.20 (dd, J 9.1, 2.8, 1H), 4.18 (q, J
7.1, 2H), 3.92-3.82 (m, 2H), 3.20-3.10 (m, 2H), 2.65-2.55 (m, 1H),
2.15-2.03 (m, 2H), 1.95-1.85 (m, 2H), 1.27 (t, J 7.1, 3H).
[0274] Intermediate 2: Ethyl
1-(6-aminopyridin-3-yl)piperidine-4-carboxylate: Intermediate 1(2.2
g, 7.9 mmol) was dissolved in EtOH (25 ml) and H.sub.2O (5 ml)
mixture. To this mixture added Iron powder (2.2 g, 39.4 mmol) and
NH.sub.4Cl (850 mg, 15.9 mmol). This mixture was stirred at
90.degree. C. for 16 h. After completion of the reaction, reaction
mixture was filtered through celite bed. Cealite bed was washed
with DCM. Filtrate was basified with aq NaHCO.sub.3 solution.
Aqueous layer was extracted with DCM. Combined DCM layers were
dried on anhydrous Na.sub.2SO.sub.4. DCM was removed on rotavapour
to obtain crude. Crude was purified by column on 60-120 mesh silica
gel using MeOH and DCM (3:97) as eluent to afford the titled
compound (1.7 g) as a brown solid. .sup.1H-NMR (.delta. ppm,
DMSO-d.sub.6, 400 MHz): 7.59 (d, J 2.8, 1H), 7.13 (dd, J 8.8, 2.8,
1H), 6.38 (d, J 8.8, 1H), 5.32 (bs, 2H), 4.07 (q, J 7.1, 2H),
3.31-3.25 (m, 2H), 2.63-2.55 (m, 2H), 2.45-2.33 (m, 1H), 1.87 (d, J
12.6, 2H), 1.73-1.62 (m, 2H), 1.18 (t, J 7.1, 3H).
[0275] Intermediate 3: Ethyl
1-(6-(2-(3-(trifluoromethoxy)phenyl)acetamido)pyridin-3-yl)piperidine-4-c-
arboxylate: Intermediate 2 (1.0 g, 3.8 mmol),
3-(Trifluoromethoxy)phenylacetic acid (1.03 g, 4.7 mmol), HATU
(1.82 g, 4.7 mmol), DIPEA (1.1 ml, 8.5 mmol) were taken in DMF (6
ml). This mixture was stirred at rt under inert atmosphere for 12
h. Reaction mass was diluted with water to obtain a solid. Solid
was filtered and dried to obtain the titled compound as a grey
solid. .sup.1H-NMR (.delta. ppm, DMSO-d.sub.6, 400 MHz): 10.43 (s,
1H), 7.99 (d, J 2.7, 1H), 7.86 (d, J 9, 1H), 7.47-7.41 (m, 1H),
7.39-7.31 (m, 3H), 7.22 (d, J 8, 1H), 4.07 (q, J 7.1, 2H), 3.73 (s,
2H), 3.56 (d, J 12.2, 2H), 2.75 (t, J 11.4, 2H), 2.50-2.41 (m, 1H),
1.90 (d, J 11.1, 2H), 1.72-1.60 (m, 2H), 1.18 (t, J 7, 3H).
[0276] Intermediate 4:
1-(6-(2-(3-(Trifluoromethoxy)phenyl)acetamido)pyridin-3-yl)piperidine-4-c-
arboxylic acid: Intermediate 3 (1.59 g, 3.5 mmol) was dissolved in
MeOH and water. To this mixture added NaOH (590 mg, 14.7 mmol).
This mixture was stirred at rt for 1 h. MeOH was removed on
rotavapour and residue was acidified with 2N HCl up to pH-5. Above
aqueous layer was extracted with MeOH and DCM (1:9) mixture.
Organic layer was dried on anhydrous Na.sub.2SO.sub.4 and
evaporated on rotavapour to obtain crude. Crude was triturated with
Et.sub.2O to obtain the titled compound as a brown solid (1.19 g).
.sup.1H-NMR (.delta. ppm, DMSO-d.sub.6, 400 MHz): 12.14 (bs, 1H),
10.43 (s, 1H), 7.99 (s, 1H), 7.86 (d, J 7.9, 1H), 7.49-7.41 (m,
1H), 7.29-7.30 (m, 3H), 7.22 (d, J 8.2, 1H), 3.73 (s, 2H), 3.56 (d,
J 11.8, 2H), 2.74 (t, J 11.1, 2H), 2.41-2.33 (m, 1H), 1.93-1.85 (m,
2H), 1.70-1.58 (m, 2H).
[0277] Intermediate 5:
N-(5-(4-(5-amino-1,3,4-thiadiazol-2-yl)piperidin-1-yl)pyridin-2-yl)-2-(3--
(trifluoromethoxy)phenyl)acetamide: Intermediate 4 (290 mg, 0.45
mmol), Thiosemicarbazide (230 mg) and POCl.sub.3 (12 ml) were mixed
and heated to 90.degree. C. for 16 h. After completion of reaction,
reaction mass was cooled to rt and quenched into crushed ice (150
g). The resulting mixture was basified to pH 14 with saturated aq.
NaOH. Aqueous layer was extracted with DCM and MeOH (9:1) mixture.
Organic layer was dried on anhydrous Na.sub.2SO.sub.4 and
evaporated on rotavapour to obtain crude (110 mg). Crude was used
in the next step without further purification. .sup.1H-NMR (.delta.
ppm, DMSO-d.sub.6, 400 MHz): 10.44 (s, 1H), 8.03 (s, 1H), 7.88 (d,
J 9, 1H), 7.48-7.30 (m, 4H), 7.22 (d, J 7.8, 1H), 6.97 (s, 2H),
3.73 (s, 2H), 3.63 (d, 12.4, 2H), 3.19-3.09 (m, 1H), 2.88 (t, J
10.9, 2H), 2.8 (d, 13.3, 2H), 1.90-1.78 (m, 2H).
[0278] Intermediate 6: Ethyl
1-(6-nitropyridin-3-yl)piperidine-3-carboxylate:
3-Bromo-6-nitropyridine (2 g, 9.85 mmol), Ethyl nipecotate (1.7 g,
10.8 mmol), K.sub.2CO.sub.3 (1.36 g, 9.84 mmol) and
tetrabutylammonium iodide (360 mg, 0.98 mmol) were taken in DMSO
(10 ml). This mixture was stirred at 100.degree. C. under inert
atmosphere for 16 hrs. After completion of the reaction, reaction
was cooled to rt and diluted with water. Aqueous layer was
extracted with EtOAc. Organic layer was dried on anhydrous
Na.sub.2SO.sub.4. EtOAc was removed on rotavapour to obtain crude.
Crude was purified on combi-flash using EtOAc and Petether (1:2) as
eluent to afford the titled compound (2.3 g) as a yellow solid.
[0279] Intermediate 7: Ethyl
1-(6-aminopyridin-3-yl)piperidine-3-carboxylate: Intermediate 6
(2.2 g, 7.9 mmol) was dissolved in EtOH (25 ml) and H.sub.2O (5 ml)
mixture. To this mixture added Iron powder (2.2 g, 39.4 mmol) and
NH.sub.4Cl (850 mg, 15.9 mmol). This mixture was stirred at
90.degree. C. for 16 h. After completion of the reaction, reaction
mixture was filtered through celite bed. Cealite bed was washed
with DCM. Filtrate was basified with aq NaHCO.sub.3 solution.
Aqueous layer was extracted with DCM. Combined DCM layers were
dried on anhydrous Na.sub.2SO.sub.4. DCM was removed on rotavapour
to obtain crude. Crude was purified by combi-flash using MeOH and
DCM (4:96) as eluent to afford the titled compound (1.8 g) as a
brown liquid. .sup.1H-NMR (.delta. ppm, DMSO-d.sub.6, 400 MHz):
7.60 (d, J 2.8, 1H), 7.13 (dd, J 8.8, 2.9, 1H), 6.38 (dd, J 8.8, 3,
1H), 5.35 (bs, 2H), 4.10 (q, J 6.8, 2H), 3.30-3.20 (m, 1H),
3.11-3.04 (m, 1H), 2.84-2.76 (m, 1H), 2.69-2.58 (m, 2H), 1.88-1.81
(m, 1H), 1.75-1.68 (m, 1H), 1.60-1.50 (m, 2H), 1.18 (t, J 7.1,
3H).
[0280] Intermediate 8: ethyl
1-(6-(2-(3-(trifluoromethoxy)phenyl)acetamido)pyridin-3-yl)piperidine-3-c-
arboxylate: Intermediate 7 (1.8 g, 7.2 mmol),
3-(Trifluoromethoxy)phenylacetic acid (1.9 g, 8.63 mmol), HATU (3.3
g, 8.7 mmol), DIPEA (3.8 ml, 21.6 mmol) were taken in DMF (5 ml).
This mixture was stirred at rt under inert atmosphere for 12 h.
Reaction mass was diluted with water and extracted with DCM. DCM
layer was dried on anhydrous Na.sub.2SO.sub.4. DCM was removed on
rotavapour to obtain the titled compound (3.2 g) which was used in
the next step without further purification. .sup.1H-NMR (.delta.
ppm, DMSO-d.sub.6, 400 MHz): 10.45 (s, 1H), 7.99 (d, J 2.6, 1H),
7.87 (d, J 9, 1H), 7.48-7.41 (m, 1H), 7.39-7.30 (m, 3H), 7.22 (d, J
8, 1H), 4.08 (q, J 7.1, 2H), 3.73 (s, 2H), 3.60-3.53 (m, 1H),
3.41-3.33 (m, 1H), 3.05-2.96 (m, 1H), 2.86-2.78 (m, 1H), 2.70-2.50
(m, 1H), 1.95-1.85 (m, 1H), 1.78-1.68 (m, 1H), 1.63-1.54 (m, 2H),
1.18 (t, J 7.1, 3H). MS (m/z): 452.6 [M+H].sup.+.
[0281] Intermediate 9:
1-(6-(2-(3-(trifluoromethoxy)phenyl)acetamido)pyridin-3-yl)piperidine-3-c-
arboxylic acid: Intermediate 8 (1.6 g, 3.5 mmol) was dissolved in
MeOH and water. To this mixture added NaOH (430 mg, 10.5 mmol).
This mixture was stirred at rt for 30 mins. MeOH was removed on
rotavapour and residue was acidified with 2N HCl up to pH.about.5.
After that above aqueous layer was extracted with MeOH and DCM
(2:8) mixture. Organic layer was dried on anhydrous
Na.sub.2SO.sub.4 and evaporated on rotavapour to obtain crude.
Crude was triturated with Et.sub.2O to obtain the titled compound
as a brown solid (950 mg). H-NMR (.delta. ppm, DMSO-d.sub.6, 400
MHz): 12.25 (s, 1H), 10.44 (s, 1H), 7.99 (d, J 2.7, 1H), 7.87 (d, J
8.8, 1H), 7.47-7.41 (m, 1H), 7.38-7.31 (m, 3H), 7.22 (d, J 7.2,
1H), 3.73 (s, 2H), 3.58 (d, J 12.4, 1H), 3.40 (d, J 11.4, 1H),
3.07-2.90 (m, 1H), 2.82-2.73 (m, 1H), 2.60-2.50 (m, 1H), 1.95-1.83
(m, 1H), 1.75-1.68 (m, 1H), 1.60-1.50 (m, 2H).
[0282] Intermediate 10:
N-(5-(3-(5-amino-1,3,4-thiadiazol-2-yl)piperidin-1-yl)pyridin-2-yl)-2-(3--
(trifluoromethoxy)phenyl)acetamide: Intermediate 9 (950 mg, 2.24
mmol), Thiosemicarbazide (610 mg, 6.7 mmol) and POCl.sub.3 (10 ml)
were mixed and heated to 90.degree. C. for 16 h. After completion
of reaction, reaction mass was cooled to rt and quenched into
crushed ice (150 g). The resulting mixture was basified to pH 14
with saturated aq. NaOH. Aqueous layer was extracted with DCM and
MeOH (9:1) mixture. Organic layer was dried on anhydrous
Na.sub.2SO.sub.4 and evaporated on rotavapour to obtain crude.
Crude was triturated with EtOAc and Petether (1:1) mixture to
obtain the titled compound (1 g) as a brown solid. MS (m/z): 479.4
[M+H].sup.+.
[0283] Intermediate 11:
1-(6-nitropyridin-3-yl)piperidine-4-carboxamide: Intermediate 1
(7.8 g, 27.9 mmol) was dissolved in MeOH (39 ml) and added aq
ammonia (46.8 ml. This mixture was heated to 50.degree. C. for 12
h. Solid that formed in the reaction mixture was filtered and dried
under vacuum to obtain the titled compound (1.7 g) as a yellow
solid. .sup.1H-NMR (.delta. ppm, DMSO-d.sub.6, 400 MHz): 8.23 (d, J
2.9, 1H), 8.11 (d, J 9.2, 1H), 7.45 (dd, J 3, 9.2, 1H), 7.25 (bs,
1H), 6.74 (bs, 1H), 4.06 (d, J 12.3, 2H), 3.10-3.00 (m, 2H),
2.45-2.35 (m, 1H), 1.81 (d, J 10.8, 2H), 1.65-1.51 (m, 2H). MS
(m/z): 250.9 [M+H].sup.+.
[0284] Intermediate 12:
1-(6-nitropyridin-3-yl)piperidine-4-carbonitrile: Intermediate 11
(1.7 g 6.79 mmol) was dissolved in Chloroform (25 ml) and added TEA
(4.8 ml, 34 mmol). This mixture was cooled to -5.degree. C. and
added trifluoroacetic anhydride (2.23 ml, 17 mmol) drop-wise. Above
mixture was stirred for 1 h at rt under N.sub.2 atmosphere.
Reaction mass was diluted with water and organic layer was
separated. Organic layer dried on anhydrous Na.sub.2SO.sub.4.
Organic layer was distilled on rotavapour to obtain the titled
compound (1.36 g) as a yellow solid. It was used in the next step
without further purification. .sup.1H-NMR (.delta. ppm,
DMSO-d.sub.6, 400 MHz): 8.25 (d, J 3, 1H), 8.13 (d, J 9.2, 1H),
7.48 (dd, J 3, 9.2, 1H), 3.80-3.70 (m, 2H), 3.42-3.33 (m, 2H),
3.20-3.11 (m, 1H), 2.05-1.96 (m, 2H), 1.85-1.75 (m, 2H).
[0285] Intermediate 13:
1-(6-aminopyridin-3-yl)piperidine-4-carbonitrile: Intermediate 12
(1.36 g, 5.85 mmol) was dissolved in EtOH (40 ml) and H.sub.2O (8
ml) mixture. To this mixture added Iron powder (1.63 g, 29.3 mmol)
and NH.sub.4Cl (624 mg, 11.7 mmol). This mixture was stirred at
90.degree. C. for 16 h. After completion of the reaction, reaction
mixture was filtered through celite bed. Cealite bed was washed
with DCM. Filtrate was basified with aq NaHCO.sub.3 solution.
Aqueous layer was extracted with DCM. Combined DCM layers were
dried on anhydrous Na.sub.2SO.sub.4. DCM was removed on rotavapour
to obtain crude. Crude was triturated with Et.sub.2O to obtain the
titled compound (1 g) as a brown solid. .sup.1H-NMR (.delta. ppm,
DMSO-d.sub.6, 400 MHz): 7.61 (d, J 2.6, 1H), 7.15 (dd, J 3, 8.8,
1H), 6.39 (d, J 8.8, 1H), 5.38 (bs, 2H), 3.10-3.03 (m, 2H),
2.99-2.91 (m, 1H), 2.86-2.78 (m, 2H), 2.00-1.93 (m, 2H), 1.85-1.75
(m, 2H).
[0286] Intermediate 14:
N-(5-(4-cyanopiperidin-1-yl)pyridin-2-yl)-2-(pyridin-2-yl)acetamide:
Intermediate 13 (440 mg, 2.29 mmol), 2-Pyridylacetic acid
hydrochloride (453 mg, 2.6 mmol), HATU (992 mg, 2.6 mmol), DIPEA
(1.1 ml, 6.5 mmol) were taken in DMF (3 ml). This mixture was
stirred at rt under inert atmosphere for 12 h. Reaction mass was
diluted with water and filtered the solid that formed. Solid was
washed with water and dried on high vacuum to obtain the titled
compound as an Off-white solid (320 mg). .sup.1H-NMR (.delta. ppm,
DMSO-d.sub.6, 400 MHz): 10.42 (s, 1H), 8.49 (d, J 4.3, 1H), 8.01
(d, J 1.8, 1H), 7.89 (d, J 9, 1H), 7.73 ((t, J 7.5, 1H), 7.41-7.35
(m, 2H), 7.27-7.21 (m, 1H), 3.87 (s, 2H), 3.35-3.28 (m, 2H),
3.07-2.98 (m, 3H), 2.03-1.94 (m, 2H), 1.86-1.77 (m, 2H).
[0287] Intermediate 15:
N-(5-(4-(5-amino-1,3,4-thiadiazol-2-yl)piperidin-1-yl)pyridin-2-yl)-2-(py-
ridin-2-yl)acetamide: Intermediate 14 (320 mg, 0.894 mmol),
Thiosemicarbazide (162 mg, 1.79 mmol) and trifluoroacetic acid (2
ml) were mixed and heated to 90.degree. C. for 2 h. After 2 h,
reaction mixture was cooled to rt and basified to pH 14 to obtain a
solid. Solid was triturated with Et.sub.2O to obtain the titled
compound (300 mg) as a brown solid. .sup.1H-NMR (.delta. ppm,
DMSO-d.sub.6, 400 MHz): 10.39 (s, 1H), 8.49 (d, J 3.2, 1H), 8.02
(d, J 2.2, 1H), 7.89 (d, J 8.7, 1H), 7.73 (t, J 7.8, 1H), 7.41-7.35
(m, 2H), 7.28-7.22 (m, 1H), 6.96 (bs, 2H), 3.87 (s, 2H), 3.67 (d, J
12.1, 2H), 3.07-2.98 (m, 1H), 2.80 (t, J 11.9, 2H), 2.03 (d, J
11.3, 2H), 1.82-1.70 (m, 2H).
[0288] Intermediate 16:
1-(6-nitropyridin-3-yl)piperidine-3-carboxamide: Intermediate 6 (5
g, 17.9 mmol) was dissolved in MeOH (25 ml) and added aq ammonia
(30 ml). This mixture was heated to 50.degree. C. for 12 h. Solid
that formed in the reaction mixture was filtered and dried under
vacuum to obtain the titled compound (1.4 g) as a yellow solid.
.sup.1H-NMR (.delta. ppm, DMSO-d.sub.6, 400 MHz): 8.24 (d, J 2.7,
1H), 8.10 (d, J 9.2, 1H), 7.46 (dd, J 2.9, 9.2, 1H), 7.35 (bs, 1H),
6.85 (bs, 1H), 4.09-3.95 (m, 2H), 3.13 (t, J 11, 1H), 3.03 (t, J
10, 1H), 2.45-2.35 (m, 1H), 1.93-1.85 (m, 1H), 1.79-1.61 (m, 2H),
1.55-1.42 (m, 1H). MS (m/z): 251.0 [M+H].sup.+.
[0289] Intermediate 17:
1-(6-nitropyridin-3-yl)piperidine-3-carbonitrile: Intermediate 16
(1.4 g 5.6 mmol) was dissolved in Chloroform (20 nil) and added TEA
(3.9 ml, 28 mmol). This mixture was cooled to -5.degree. C. and
added trifluoroacetic anhydride (1.95 ml, 14 mmol) drop-wise. Above
mixture was stirred for 1 h at rt under N.sub.2 atmosphere.
Reaction mass was diluted with water and organic layer was
separated. Organic layer dried on anhydrous Na.sub.2SO.sub.4.
Organic layer was distilled on rotavapour to obtain the titled
compound (1g) as a yellow solid. It was used in the next step
without further purification. .sup.1H-NMR (.delta. ppm,
DMSO-d.sub.6, 400 MHz): 8.30 (d, J 2.9, 1H), 8.13 (d, J 9.2, 1H),
7.54 (dd, J 3, 9.2, 1H), 3.88-3.75 (m, 2H), 3.62-3.45 (m, 2H),
3.17-3.09 (m, 1H), 2.01-1.75 (m, 2H), 1.70-1.58 (m, 2H). MS (m/z):
232.9 [M+H].sup.+.
[0290] Intermediate 18:
1-(6-aminopyridin-3-yl)piperidine-3-carbonitrile: Intermediate 17
(1 g, 4.3 mmol) was dissolved in EtOH (30 ml) and H.sub.2O (6 ml)
mixture. To this mixture added Iron powder (1.20 g, 21.5 mmol) and
NH.sub.4Cl (460 mg, 8.7 mmol). This mixture was stirred at
90.degree. C. for 16 h. After completion of the reaction, reaction
mixture was filtered through celite bed. Cealite bed was washed
with DCM. Filtrate was basified with aq NaHCO.sub.3 solution.
Aqueous layer was extracted with DCM. Combined DCM layers were
dried on anhydrous Na.sub.2SO.sub.4. DCM was removed on rotavapour
to obtain crude. Crude was purified by combi-flash using MeOH and
DCM (3:97) as eluent to afford the titled compound (880 mg) as a
black gel.
[0291] Intermediate 19:
N-(5-(3-cyanopiperidin-1-yl)pyridin-2-yl)-2-(pyridin-2-yl)acetamide:
Intermediate 18 (440 mg, 2.2 mmol), 2-Pyridylacetic acid
hydrochloride (453 mg, 2.6 mmol), HATU (992 mg, 2.6 mmol), DIPEA
(1.1 ml, 6.5 mmol) were taken in DMF (3 ml). This mixture was
stirred at rt under inert atmosphere for 12 h. Reaction mass was
diluted with water and extracted with DCM. DCM was dried on
anhydrous Na.sub.2SO.sub.4 and DCM was removed on rotavapour to
obtain crude. Crude was purified by combi-flash using MeOH and DCM
(3:97) as eluent to afford the titled compound (450 mg) as a brown
solid. .sup.1H-NMR (.delta. ppm, DMSO-d.sub.6, 400 MHz): 10.45 (s,
1H), 8.49 (d, J 4, 1H), 8.03 (d, J 2.5, 1H), 7.90 (d, J 8.4, 1H),
7.73 (t, J 6.3, 1H), 7.44-7.33 (m, 2H), 7.27-7.21 (m, 1H), 3.87 (s,
2H), 3.40-3.31 (m, 2H), 3.20-3.04 (m, 3H), 1.90-1.71 (m, 3H),
1.70-1.59 (m, 1H). MS (m/z): 322.0 [M+H].sup.+.
[0292] Intermediate 20: N-(5-(3-(5-amino-1, 3,
4-thiadiazol-2-yl)piperidin-1-yl)pyridin-2-yl)-2-(pyridin-2-yl)acetamide:
Intermediate 19 (425 mg, 1.87 mmol), Thiosemicarbazide (216 mg,
2.37 mmol) and trifluoroacetic acid (2 ml) were mixed and heated to
90.degree. C. for 2 h. After 2 h, reaction mixture was cooled to rt
and basified to pH 14. Aqueous layer was extracted with MeOH and
DCM (1:9) mixture. Organic layer was dried on anhydrous
Na.sub.2SO.sub.4 and removed MeOH and DCM on rotavapour to obtain
crude. Crude was purified by combi-flash using MeOH and DCM (7:93)
as eluent to afford the titled compound (164 mg) as a pink solid.
MS (m/z): 396.1 [M+H].sup.+.
[0293] Intermediate 21: Ethyl
1-(6-aminopyridazin-3-yl)piperidine-4-carboxylate:
3-Amino-6-chloropyridazine (1 g, 7.72 mmol) and Ethyl isonipecotate
(2.4 g, 15.39) mmol were mixed and heated to 180.degree. C. for 6
h. After 6 h, reaction mass cooled to rt and added satd. Aqueous
NaHCO.sub.3 solution (50 ml). This mixture was extracted with DCM.
DCM removed on rotavapour to obtain crude. Crude was purified by
combi-flash using MeOH and DCM (3:97) as eluent to afford the
titled compound as a brown solid (1.5 g).
[0294] Intermediate 22: Ethyl
1-(6-(2-(3-(trifluoromethoxy)phenyl)acetamido)pyridazin-3-yl)piperidine-4-
-carboxylate: Intermediate 21 (1.5 g, 5.99 mmol),
3-(Trifluoromethoxy)phenylacetic acid (1.58 g, 7.17 mmol), HATU (5
g, 13.14 mmol), DIPEA (3.1 ml, 17.78 mmol) were taken in DMF (4
ml). This mixture was stirred at rt under inert atmosphere for 12
h. Reaction mass was diluted with water and extracted with DCM. DCM
layer was dried on anhydrous Na.sub.2SO.sub.4 and DCM removed on
rotavapour to obtain crude. Crude was purified by combi-flash using
MeOH and DCM (1:99) as eluent to afford the titled compound as a
brown gummy solid (1.1 g). .sup.1H-NMR (.delta. ppm, DMSO-d.sub.6,
400 MHz): 10.93 (s, 1H), 7.98 (d, J 9.8, 1H), 7.48-7.42 (m, 1H),
7.38-7.30 (m, 3H), 7.24 (d, J 8, 1H), 4.15 (d, J 13.3, 2H), 4.06
(q, J 7.1, 2H), 3.78 (s, 2H), 2.98 (t, J 11.4, 2H), 2.63-2.55 (m,
1H), 1.89 (d, J 10.6, 2H), 1.62-1.50 (m, 2H), 1.17 (t, J 7.1,
3H).
[0295] Intermediate 23:
1-(6-(2-(3-(trifluoromethoxy)phenyl)acetamido)pyridazin-3-yl)
piperidine-4-carboxylic acid: Intermediate 22 (1.1 g, 2.43 mmol)
was dissolved in MeOH and water. To this mixture added NaOH (290
mg, 7.25 mnol). This mixture was stirred at rt for 16 h. Reaction
mass was acidified with dil. HCl up to pH.about.5 to obtain a
solid. Solid was filtered and dried to obtain the titled compound
as a yellow solid (450 mg). .sup.1H-NMR (.delta. ppm, DMSO-d.sub.6,
400 MHz): 12.27 (bs, 1H), 10.92 (s, 1H), 7.97 (d, J 9.8, 1H),
7.47-7.42 (m, 1H), 7.37-7.29 (m, 3H), 7.28-7.20 (m, 1H), 4.14 (d, J
13.2, 2H), 3.78 (s, 2H), 3.40-3.30 (m, 1H), 3.01-2.93 (m, 2H), 1.87
(d, J 10.6, 2H), 1.60-1.49 (m, 2H). MS (m/z): 425.0
[M+H].sup.+.
[0296] Intermediate 24:
N-(6-(4-(5-amino-1,3,4-thiadiazol-2-yl)piperidin-1-yl)pyridazin-3-yl)-2-(-
3-(trifluoromethoxy)phenyl)acetamide: Intermediate 23 (510 mg, 1.20
mmol), Thiosemicarbazide (330 mg, 3.6 mmol) and POCl.sub.3 (5 ml)
were mixed and heated to 90.degree. C. for 3 h. After completion of
reaction, reaction mass was cooled to rt and quenched into crushed
ice (150 g). The resulting mixture was basified to pH 10 with
saturated aq. NaOH. Aqueous layer was extracted with DCM and MeOH
(9:1) mixture. Organic layer was dried on anhydrous
Na.sub.2SO.sub.4 and evaporated on rotavapour to obtain crude.
Crude was purified by combi-flash using MeOH and DCM (6:94) as
eluent to afford the titled compound as a pale-yellow solid (100
mg). .sup.1H-NMR (.delta. ppm, DMSO-d.sub.6, 400 MHz): 10.93 (s,
1H), 7.98 (d, J 9.8, 1H), 7.48-7.42 (m, 1H), 7.37-7.32 (m, 3H),
7.24 (d, J 8, 1H), 7.00 (s, 2H), 4.27 (d, J 13.3, 2H), 3.78 (s,
2H), 3.20-3.10 (m, 1H), 3.01 (t, J 11.6, 2H), 2.02 (d, J 10.8, 2H),
1.70-1.58 (m, 2H). MS (m/z): 479.8 [M+H].sup.+.
[0297] Intermediate 25: ethyl
1-(6-aminopyridazin-3-yl)piperidine-3-carboxylate:
3-Amino-6-chloropyridazine (3 g, 23.2 mmol) and Ethyl nipecotate
(7.3 g, 46.4) mmol were mixed and heated to 180.degree. C. for 6 h.
After 6 h, reaction mass cooled to rt and added satd. Aqueous
NaHCO.sub.3 solution (50 ml). This mixture was extracted with DCM.
DCM removed on rotavapour to obtain crude. Crude was purified by
column chromatography on 60-120 mesh silica gel using MeOH and DCM
(3:97) as eluent to afford the titled compound as a brown gummy
solid (2.9 g). MS (m/z): 250.8 [M+H].sup.+.
[0298] Intermediate 26: Ethyl
1-(6-(2-(3-(trifluoromethoxy)phenyl)acetamido)pyridazin-3-yl)piperidine-3-
-carboxylate: Intermediate 25 (2.9 g, 11.6 mmol),
3-(Trifluoromethoxy)phenylacetic acid (3.06 g, 13.9 mmol), HATU
(9.7 g, 25.5 mmol), DIPEA (2 ml, 34.75 mmol) were taken in DMF (6
ml). This mixture was stirred at rt under inert atmosphere for 12
h. Reaction mass was diluted with water and extracted with DCM. DCM
layer was dried on anhydrous Na.sub.2SO.sub.4 and DCM removed on
rotavapour to obtain crude. Crude was purified by column
chromatography on 60-120 mesh silica gel using MeOH and DCM (1:99)
as eluent to afford the titled compound as a brown solid (3.1 g).
.sup.1H-NMR (.delta. ppm, DMSO-d.sub.6, 400 MHz): 10.91 (s, 1H),
7.97 (d, J 9.8, 1H), 7.47-7.43 (m, 1H), 7.37-7.31 (m, 3H), 7.24 (d,
J 8, 1H), 4.25 (d, J 13, 1H), 4.06 (q, J 7, 2H), 3.90 (d, J 12.9,
1H), 3.78 (s, 2H), 3.21-3.05 (m, 2H), 2.60-2.51 (m, 1H), 2.00-1.91
(m, 1H), 1.72-1.53 (m, 2H), 1.52-1.41 (m, 1H), 1.17 (t, J 7.1,
3H).
[0299] Intermediate 27:
1-(6-(2-(3-(Trifluoromethoxy)phenyl)acetamido)pyridazin-3-yl)
piperidine-3-carboxylic acid: Intermediate 26 (3.1 g, 6.85 mmol)
was dissolved in MeOH and water. To this mixture added NaOH (1.64
g, 41.1 mmol). This mixture was stirred at rt for 3 h. Reaction
mass was acidified with dil. HCl up to pH.about.5 to obtain a
solid. Solid was filtered and dried to obtain the titled compound
as a yellow solid (1.4 g). .sup.1H-NMR (.delta. ppm, DMSO-d.sub.6,
400 MHz): MS (m/z): 425.0 [M+H].sup.+. 12.33 (bs, 1H), 10.92 (s,
1H), 7.97 (d, J 9.7, 1H), 7.48-7.42 (m, 1H), 7.38-7.31 (m, 3H),
7.24 (d, J 7.6, 1H), 4.25 (d, J 11.3, 1H), 3.95 (d, J 13.3, 1H),
3.78 (s, 2H), 3.15-3.00 (m, 2H), 2.00-1.90 (m, 1H), 1.75-1.60 (m,
2H), 1.55-1.40 (m, 1H). MS (m/z): 425.0 [M+H].sup.+.
[0300] Intermediate 28:
N-(6-(3-(5-amino-1,3,4-thiadiazol-2-yl)piperidin-1-yl)pyridazin-3-yl)-2-(-
3-(trifluoromethoxy)phenyl)acetamide: Intermediate 27 (1.4 g, 3.3
mmol), Thiosemicarbazide (900 mg, 9.9 mmol) and POCl.sub.3 (14 ml)
were mixed and heated to 90.degree. C. for 3 h. After completion of
reaction, reaction mass was cooled to rt and quenched into crushed
ice. The resulting mixture was basified to pH 10 with saturated aq.
NaOH. Aqueous layer was extracted with DCM and MeOH (9:1) mixture.
Organic layer was dried on anhydrous Na.sub.2SO.sub.4 and
evaporated on rotavapour to obtain crude. Crude was purified by
combi-flash using MeOH and DCM (6:94) as eluent to afford the
titled compound as a white solid (180 mg). .sup.1H-NMR (.delta.
ppm, DMSO-d.sub.6, 400 MHz): 10-94 (s, 1H), 7.98 (d, J 9.5, 1H),
7.48-7.42 (m, 1H), 7.41-7.32 (m, 3H), 7.24 (d, J 7.7, 1H), 7.04 (s,
2H), 4.38 (d, J 12.2, 1H), 4.02 (d, J 12.4, 1H), 3.78 (s, 2H),
3.23-3.05 (m, 3H), 2.11-2.03 (m, 1H), 1.80-1.70 (m, 2H), 1.65-1.53
(m, 1H). MS (m/z): 480.4 [M+H].sup.+.
[0301] Intermediate 29: Ethyl
1-(6-(2-(2-chlorophenyl)acetamido)pyridazin-3-yl)piperidine-4-carboxylate-
: Intermediate 21 (1.35 g, 5.4 mmol), 2-Chlorophenylacetic acid
(1.11 g, 6.5 mmol), HATU (4.5 g, 11.85 mmol), DIPEA (2.8 ml, 16.2
mmol) were taken in DMF (4 ml). This mixture was stirred at rt
under inert atmosphere for 12 h. Reaction mass was diluted with
water and extracted with DCM. DCM layer was dried on anhydrous
Na.sub.2SO.sub.4 and DCM removed on rotavapour to obtain crude.
Crude was purified by column chromatography on 60-120 mesh silica
gel using MeOH and DCM (1:99) as eluent to afford the titled
compound as a brown gummy solid (510 mg). .sup.1H-NMR (.delta. ppm,
DMSO-d.sub.6, 400 MHz): 10.91 (s, 1H), 7.97 (d, J 9.7, 1H),
7.45-7.38 (m, 2H), 7.36-7.25 (m, 3H), 4.20-4.13 (m, 2H), 4.06 (q, J
7.1, 2H), 3.90 (s, 2H), 2.99 (t, J 11.1, 2H), 2.64-2.56 (m, 1H),
1.93-1.85 (m, 2H), 1.62-1.51 (m, 2H), 1.17 (t, J 7.1, 3H).
[0302] Intermediate 30:
1-(6-(2-(2-Chlorophenyl)acetamido)pyridazin-3-yl)piperidine-4-carboxylic
acid: Intermediate 29 (510 mg, 1.26 mmol) was dissolved in MeOH and
water. To this mixture added NaOH (302 mg, 7.6 mmol). This mixture
was stirred at rt for 6 h. Reaction mass was acidified with dil.
HCl up to pH.about.5 to obtain a solid. Solid was filtered and
dried to obtain the titled compound as a yellow solid (350 mg).
.sup.1H-NMR (.delta. ppm, DMSO-d.sub.6, 400 MHz): 12.22 (s, 1H),
10.90 (s, 1H), 7.96 (d, J 9.5, 1H), 7.45-7.38 (m, 2H), 7.36-7.25
(m, 3H), 4.14 (d, J 13.2, 2H), 3.90 (s, 2H), 2.98 (t, J 11.4, 2H),
2.55-2.50 (m, 1H), 1.87 (d, J 11.2, 2H), 1.61-1.50 (m, 2H).
[0303] Intermediate 31:
N-(6-(4-(5-Amino-1,3,4-thiadiazol-2-yl)piperidin-1-yl)pyridazin-3-yl)-2-(-
2-chlorophenyl)acetamide: Intermediate 30 (350 mg, 0.93 mmol),
Thiosemicarbazide (255 mg, 2.3 mmol) and POCl.sub.3 (3.5 ml) were
mixed and heated to 90.degree. C. for 3 h. After completion of
reaction, reaction mass was cooled to rt and quenched into crushed
ice (150 g). The resulting mixture was basified to pH 10 with
saturated aq. NaOH. Aqueous layer was extracted with DCM and MeOH
(9:1) mixture. Organic layer was dried on anhydrous
Na.sub.2SO.sub.4 and evaporated on rotavapour to obtain crude.
Crude was purified by combi-flash using MeOH and DCM (6:94) as
eluent to afford the titled compound as a brown solid (40 mg).
.sup.1H-NMR (.delta. ppm, DMSO-d.sub.6, 400 MHz): 10.91 (s, 1H),
7.98 (d, J 9.4, 1H), 7.46-7.35 (m, 3H), 7.31-7.25 (m, 2H), 7.01 (s,
2H), 4.28 (d, J 12.9, 2H), 3.90 (s, 2H), 3.20-3.11 (m, 1H), 3.10
(t, J 11.4, 2H), 2.02 (d, J 11.6, 2H), 1.70-1.59 (m, 2H).
[0304] Intermediate 32:
1-(6-aminopyridazin-3-yl)piperidine-4-carbonitrile:
3-Amino-6-chloropyridazine (3 g, 23.2 mmol) and 4-Cyanopiperidine
(3.8 g, 34.7) mmol were mixed and heated to 180.degree. C. for 4 h.
After 4 h, reaction mass cooled to rt and hard reaction mass
dissolved in MeOH and DCM (1:9) mixture. MeOH and DCM were removed
on rotavapour to obtain crude. Crude was purified by column
chromatography on 60-120 mesh silica gel to obtain the titled
compound (4.6 g) as a dark red solid.
[0305] Intermediate 33:
N-(6-(4-cyanopiperidin-1-yl)pyridazin-3-yl)-2-(2-fluorophenyl)acetamide:
Intermediate 32 (365 mg, 1.8 mmol), 2-Fluorophenylacetic acid (388
mg, 2.5 mmol), HATU (1.5 g, 3.95 mmol), DIPEA (0.9 ml, 5.4 mmol)
were taken in DMF (3 ml). This mixture was stirred at rt under
inert atmosphere for 1 h. Reaction mass was diluted with water to
obtain a solid. Solid was dried on high vacuum to obtain the titled
compound (80 mg) as a brown solid. .sup.1H-NMR (.delta. ppm,
DMSO-d.sub.6, 400 MHz): 10.93 (s, 1H), 7.99 (d, J 9.8, 1H),
7.40-7.25 (m, 3H), 7.19-7.11 (m, 2H), 3.85-3.75 (m, 4H), 3.41-3.35
(m, 2H), 3.16-3.05 (m, 1H), 1.98-1.90 (m, 2H), 1.80-1.70 (m,
2H).
[0306] Intermediate 34:
N-(6-(4-(5-amino-1,3,4-thiadiazol-2-yl)piperidin-1-yl)pyridazin-3-yl)-2-(-
2-fluorophenyl)acetamide: Intermediate 33 (80 mg, 0.24 mmol),
Thiosemicarbazide (43 mg, 0.47 mmol) and trifluoroacetic acid (1
ml) were mixed and heated to 90.degree. C. for 3 h. After
completion of reaction, reaction mass was cooled to rt and basified
to pH 14 with saturated aq. NaOH to obtain a solid. Solid was
filtered and dried on high vacuum to obtain the titled compound (60
mg) as a pale-brown solid. 1H-NMR (.delta. ppm, DMSO-d6, 400 MHz):
10.91 (s, 1H), 7.98 (d, J 10, 1H), 7.40-7.26 (m, 3H), 7.19-7.12 (m,
2H), 7.01 (s, 2H), 4.28 (d, J 12.6, 2H), 3.80 (s, 2H), 3.20-3.10
(m, 1H), 3.01 (t, J 12.2, 2H), 2.03 (d, J 11.6, 2H), 1.70-1.58 (m,
2H).
[0307] Intermediate 35:
N-(6-(4-cyanopiperidin-1-yl)pyridazin-3-yl)-2-(pyridin-2-yl)acetamide:
Intermediate 32 (500 mg, 2.5 mmol), 2-Pyridineacetic acid
hydrochloride (500 mg, 2.95 mmol), HATU (2.05 g, 5.41 mmol), DIPEA
(1.27 ml, 7.38 mmol) were taken in DMF (3 ml). This mixture was
stirred at rt under inert atmosphere for 1 h. Reaction mass was
diluted with water to obtain a solid. Solid was dried on high
vacuum to obtain the titled compound (370 mg) as a brown solid. MS
(m/z): 322.26 [M+H].sup.+.
[0308] Intermediate 36:
N-(6-(4-(5-amino-1,3,4-thiadiazol-2-yl)piperidin-1-yl)pyridazin-3-yl)-2-(-
pyridin-2-yl)acetamide: Intermediate 35 (360 mg, 1.1 mmol),
Thiosemicarbazide (203 mg, 2.22 mmol) and trifluoroacetic acid (4
ml) were mixed and heated to 90.degree. C. for 3 h. After
completion of reaction, reaction mass was cooled to rt and basified
to pH 14 with saturated aq. NaOH to obtain a solid. Solid was
filtered and dried on high vacuum to obtain the titled compound
(180 mg) as a pale-brown solid.
[0309] Intermediate 37:
N-(6-(4-cyanopiperidin-1-yl)pyridazin-3-yl)-2-(pyridin-3-yl)acetamide:
Intermediate 32 (500 mg, 2.5 mmol), 3-Pyridineacetic acid
hydrochloride (500 mg, 2.95 mmol), HATU (2.05 g, 5.41 mmol), DIPEA
(1.27 ml, 7.38 mmol) were taken in DMF (3 ml). This mixture was
stirred at rt under inert atmosphere for 1 h. Reaction mass was
diluted with water to obtain a solid. Solid was purified by
combi-flash using MeOH and DCM (5:95) as eluent to afford the
titled compound (180 mg) as a brown solid. MS (m/z): 322.8
[M+H].sup.+.
[0310] Intermediate 38:
N-(6-(4-(5-amino-1,3,4-thiadiazol-2-yl)piperidin-1-yl)pyridazin-3-yl)-2-(-
pyridin-3-yl)acetamide: Intermediate 37 (180 mg, 0.55 mmol),
Thiosemicarbazide (101 mg, 1.1 mmol) and trifluoroacetic acid (3
ml) were mixed and heated to 90.degree. C. for 3 h. After
completion of reaction, reaction mass was cooled to rt and basified
to pH 14 with saturated aq. NaOH to obtain a solid. Solid was
filtered and dried on high vacuum to obtain the titled compound (60
mg) as a pale-brown solid.
[0311] Intermediate 39:
2-(1-(6-aminopyridazin-3-yl)piperidin-4-yl)acetonitrile:
3-Amino-6-chloropyridazine (350 mg, 2.70 mmol) and
2-(piperidin-4-yl)acetonitrile (670 mg, 5.4 mmol) were mixed and
heated to 180.degree. C. for 4 h. After 4 h, reaction mass cooled
to rt and hard reaction mass dissolved in MeOH and DCM (1:9)
mixture. MeOH and DCM were removed on rotavapour to obtain crude.
Crude was purified by column chromatography on 60-120 mesh silica
gel to obtain the titled compound (300 mg) as a brown gummy
solid.
[0312] Intermediate 40:
N-(6-(4-(cyanomethyl)piperidin-1-yl)pyridazin-3-yl)-2-(3-(trifluoromethox-
y)phenyl)acetamide: Intermediate 39 (300 mg, 1.38 mmol),
3-(Trifluoromethoxy)phenylacetic acid (360 mg, 1.63 mmol), HATU
(1.2 g, 3.16 mmol), DIPEA (0.73 ml, 4.2 mmol) were taken in DMF (3
ml). This mixture was stirred at rt under inert atmosphere for 1 h.
Reaction mass was diluted with water to obtain a solid. Solid was
purified by column chromatography using MeOH and DCM (2:98) as
eluent to afford the titled compound (50 mg) as a brown solid.
.sup.1H-NMR (.delta. ppm, DMSO-d.sub.6, 400 MHz): 10.91 (s, 1H),
7.96 (d, J 9.7, 1H), 7.48-7.42 (m, 1H), 7.38-7.30 (m, 3H), 7.23 (d,
J 8.1, 1H), 4.27 (d, J 13.2, 2H), 2.86 (t, J 11.7, 2H), 2.55-2.50
(m, 2H), 1.95-1.85 (m, 1H), 1.77 (d, J 12.9, 2H), 1.73-1.58 (m,
2H).
[0313] Intermediate 41:
N-(6-(4-((5-amino-1,3,4-thiadiazol-2-yl)methyl)piperidin-1-yl)pyridazin-3-
-yl)-2-(3-(trifluoromethoxy)phenyl)acetamide: Intermediate 40 (50
mg, 0.12 mmol), Thiosemicarbazide (22 mg, 0.24 mmol) and
trifluoroacetic acid (2 ml) were mixed and heated to 90.degree. C.
for 12 h. After completion of reaction, reaction mass was cooled to
rt and basified to pH 14 with saturated aq. NaOH to obtain a solid.
Solid was filtered and dried on high vacuum to obtain the titled
compound (34 mg) as a brown solid. MS (m/z): 494.1 [M+H].sup.+.
Example 1
2-(Pyridin-2-yl)-N-(5-(1-(6-(2-(3-(trifluoromethoxy)phenyl)acetamido)pyrid-
in-3-yl)piperidin-4-yl)-1,3,4-thiadiazol-2-yl)acetamide
[0314] Intermediate 5 (100 mg, 0.21 mmol), 2-Pyridylacetic acid
hydrochloride (44 mg, 0.25 mmol), HATU (96 mg, 0.25 mmol),
N-Ethyldiisopropyl amine (0.1 ml, 0.62 mmol) were dissolved in DMF
(2 ml). This mixture was stirred at rt for 30 mins. Reaction mass
was diluted with water and extracted with EtOAc. Organic layer was
washed with water. Organic layer was dried on anhydrous
Na.sub.2SO.sub.4. EtOAc was removed on rotavapour to obtain crude.
Crude was purified by column chromatography on 60-120 mesh silica
gel using Methanol and DCM (3:97) as eluent to afford the titled
compound (15 mg) as a brown solid. M.P.: 195-197.degree. C.
.sup.1H-NMR (.delta. ppm, DMSO-d.sub.6, 400 MHz): 12.64 (s, 1H),
10.44 (s, 1H), 8.47 (d, J 4.2, 1H), 8.03 (d, J 2.7, 1H), 7.87 (d, J
9, 1H), 7.75 (t, J 7.7, 1H), 7.48-7.32 (m, 5H), 7.30-7.20 (m, 2H),
4.00 (s, 2H), 3.73 (s, 2H), 3.72-3.65 (m, 2H), 3.30-3.20 (m, 1H),
2.84 (t, J 11.7, 2H), 2.11 (d, J 11.5, 2H), 1.90-1.78 (m, 2H).
Example 2
(RS)-2-(Pyridin-2-yl)-N-(5-(1-(6-(2-(3-(trifluoromethoxy)phenyl)acetamido)-
pyridin-3-yl) piperidin-3-yl)-1,3,4-thiadiazol-2-yl)acetamide
[0315] Intermediate 10 (500 mg, 1.04 mmol), 2-Pyridylacetic acid
hydrochloride (220 mg, 1.27 mmol), HATU (480 mg, 1.27 mmol),
N-Ethyldiisopropyl amine (0.5 ml, 3.09 mmol) were dissolved in DMF
(3 ml). This mixture was stirred at rt for 30 mins. Reaction mass
was diluted with water and extracted with DCM. Organic layer was
washed with water. Organic layer was dried on anhydrous
Na.sub.2SO.sub.4. DCM was removed on rotavapour to obtain crude.
Crude was purified by column chromatography by combi-flash using
Methanol and DCM (4:96) as eluent to afford the titled compound (10
mg) as a brown solid. M.P.: 187-190.degree. C. .sup.1H-NMR (.delta.
ppm, DMSO-d.sub.6, 400 MHz): 12.63 (s, 1H), 10.46 (s, 1H), 8.48 (d,
J 4, 1H), 8.04 (d, J 2.6, 1H), 7.88 (d, J 9, 1H), 7.75 (d, J 7.6,
1H), 7.46-7.30 (m, 5H), 7.29-7.20 (m, 2H), 3.99 (s, 2H), 3.79-3.69
(m, 3H), 3.48-3.40 (m, 1H), 3.15-3.08 (m, 1H), 3.00-2.90 (m, 1H),
2.30-2.20 (m, 1H), 2.15-2.05 (m, 1H), 1.82-1.70 (m, 3H).
Example 2A
(R) or (S)
2-(Pyridin-2-yl)-N-(5-(1-(6-(2-(3-(trifluoromethoxy)phenyl)acet-
amido)
pyridin-3-yl)piperidin-3-yl)-1,3,4-thiadiazol-2-yl)acetamide
[0316] The enantiomerically pure isomer was separated by
preparative SFC conditions from
2-(Pyridin-2-yl)-N-(5-(1-(6-(2-(3-(trifluoromethoxy)phenyl)acetamido)pyri-
din-3-yl)piperidin-3-yl)-1,3,4-thiadiazol-2-yl)acetamide (Example:
2) (0.300 g) on a CHIRALPAK IC 4.6*250, 5 um (Daicel) using
n-Hexane (0.1% DEA)/Ethanol (0.1% DEA)=40/60 as the mobile phase at
a flow rate of 1.0 ml/min to obtain the titled compound (55 mg) as
a brown solid e.e. 100%. Rt: 12.34 min. M.P.: 122-124.degree. C.
.sup.1H-NMR (.delta. ppm, DMSO-d.sub.6, 400 MHz): 12.63 (s, 1H),
10.46 (s, 1H), 8.48 (d, J 4, 1H), 8.04 (d, J 2.6, 1H), 7.88 (d, J
9, 1H), 7.75 (d, J 7.6, 1H), 7.46-7.30 (m, 5H), 7.29-7.20 (m, 2H),
3.99 (s, 2H), 3.79-3.69 (m, 3H), 3.48-3.40 (m, 1H), 3.15-3.08 (m,
1H), 3.00-2.90 (m, 1H), 2.30-2.20 (m, 1H), 2.15-2.05 (m, 1H),
1.82-1.70 (m, 3H).
Example 2B
(S) or (R)
2-(Pyridin-2-yl)-N-(5-(1-(6-(2-(3-(trifluoromethoxy)phenyl)acet-
amido)
pyridin-3-yl)piperidin-3-yl)-1,3,4-thiadiazol-2-yl)acetamide
[0317] The enantiomerically pure isomer was separated by
preparative SFC conditions from
2-(Pyridin-2-yl)-N-(5-(1-(6-(2-(3-(trifluoromethoxy)phenyl)acetamido)pyri-
din-3-yl)piperidin-3-yl)-1,3,4-thiadiazol-2-yl)acetamide (Example:
2) (0.300 g) on a CHIRALPAK IC 4.6*250, 5 um (Daicel) using
n-Hexane (0.1% DEA)/Ethanol (0.1% DEA)=40/60 as the mobile phase at
a flow rate of 1.0 ml/min to obtain the titled compound (45 mg) as
a brown solid e.e. 100%. Rt: 14.47 min. M.P.: 129-131.degree. C.
.sup.1H-NMR (.delta. ppm, DMSO-d.sub.6, 400 MHz): 12.63 (s, 1H),
10.46 (s, 1H), 8.48 (d, J 4, 1H), 8.04 (d, J 2.6, 1H), 7.88 (d, J
9, 1H), 7.75 (d, J 7.6, 1H), 7.46-7.30 (m, 5H), 7.29-7.20 (m, 2H),
3.99 (s, 2H), 3.79-3.69 (m, 3H), 3.48-3.40 (m, 1H), 3.15-3.08 (m,
1H), 3.00-2.90 (m, 1H), 2.30-2.20 (m, 1H), 2.15-2.05 (m, 1H),
1.82-1.70 (m, 3H).
Example 3
(RS)-2-(Pyridin-3-yl)-N-(5-(1-(6-(2-(3-(trifluoromethoxy)phenyl)acetamido)-
pyridin-3-yl) piperidin-3-yl)-1,3,4-thiadiazol-2-yl)acetamide
[0318] Intermediate 10 (500 mg, 1.04 mmol), 3-Pyridylacetic acid
hydrochloride (220 mg, 1.26 mmol), HATU (480 mg, 1.25 mmol),
N-Ethyldiisopropyl amine (0.5 ml, 3.1 mmol) were dissolved in DMF
(3 ml). This mixture was stirred at rt for 30 mins. Reaction mass
was diluted with water and extracted with DCM. Organic layer was
washed with water. Organic layer was dried on anhydrous
Na.sub.2SO.sub.4. DCM was removed on rotavapour to obtain crude.
Crude was purified by combi-flash using Methanol and DCM (5:95) as
eluent to afford the titled compound (6 mg) as a Pale-Yellow solid.
M.P.: 145-147.degree. C. .sup.1H-NMR (.delta. ppm, DMSO-d.sub.6,
400 MHz): 12.69 (s, 1H), 10.47 (s, 1H), 8.55-8.51 (m, 2H), 8.03 (s,
1H), 7.90-7.80 (m, 2H), 7.49-7.40 (m, 3H), 7.30-7.27 (m, 2H), 7.22
(d, J 8, 1H), 3.89 (s, 2H), 3.75-3.70 (m, 3H), 3.20-3.10 (m, 2H),
3.05-2.92 (m, 2H), 2.13-2.04 (m, 1H), 1.80-1.60 (m, 3H).
Example 3A
(R) or (S)
2-(Pyridin-3-yl)-N-(5-(1-(6-(2-(3-(trifluoromethoxy)phenyl)acet-
amido)
pyridin-3-yl)piperidin-3-yl)-1,3,4-thiadiazol-2-yl)acetamide
[0319] The enantiomerically pure isomer was separated by
preparative SFC conditions from
2-(Pyridin-3-yl)-N-(5-(1-(6-(2-(3-(trifluoromethoxy)phenyl)acetamido)pyri-
din-3-yl)piperidin-3-yl)-1,3,4-thiadiazol-2-yl)acetamide (Example:
3) (460 mg) on a CHIRALPAK IC 4.6*250, 5 um (Daicel) using n-Hexane
(0.1% DEA)/Ethanol (0.1% DEA)=50/50 as the mobile phase at a flow
rate of 1.0 ml/min to obtain the titled compound (100 mg) as a
brown solid e.e. 100%. Rt: 12.11 min. M.P.: 170-172.degree. C.
.sup.1H-NMR (.delta. ppm, DMSO-d.sub.6, 400 MHz): 12.69 (s, 1H),
10.47 (s, 1H), 8.55-8.51 (m, 2H), 8.03 (s, 1H), 7.90-7.80 (m, 2H),
7.49-7.40 (m, 3H), 7.30-7.27 (m, 2H), 7.22 (d, J 8, 1H), 3.89 (s,
2H), 3.75-3.70 (m, 3H), 3.20-3.10 (m, 2H), 3.05-2.92 (m, 2H),
2.13-2.04 (m, 1H), 1.80-1.60 (m, 3H).
Example 3B
(S) or (R)
2-(Pyridin-3-yl)-N-(5-(1-(6-(2-(3-(trifluoromethoxy)phenyl)acet-
amido)
pyridine-3-yl)piperidin-3-yl)-1,3,4-thiadiazol-2-yl)acetamide
[0320] The enantiomerically pure isomer was separated by
preparative SFC conditions from
2-(Pyridin-3-yl)-N-(5-(1-(6-(2-(3-(trifluoromethoxy)phenyl)acetamido)pyri-
din-3-yl)piperidin-3-yl)-1,3,4-thiadiazol-2-yl)acetamide (Example:
3) (460 mg) on a CHIRALPAK IC 4.6*250, 5 um (Daicel) using n-Hexane
(0.1% DEA)/Ethanol (0.1% DEA)=50/50 as the mobile phase at a flow
rate of 1.0 ml/min to obtain the titled compound (100 mg) as a
brown solid e.e. 100%. Rt: 14.12 min. M.P.: 141-143.degree. C.
.sup.1H-NMR (.delta. ppm, DMSO-d.sub.6, 400 MHz): 12.69 (s, 1H),
10.47 (s, 1H), 8.55-8.51 (m, 2H), 8.03 (s, 1H), 7.90-7.80 (m, 2H),
7.49-7.40 (m, 3H), 7.30-7.27 (m, 2H), 7.22 (d, J 8, 1H), 3.89 (s,
2H), 3.75-3.70 (m, 3H), 3.20-3.10 (m, 2H), 3.05-2.92 (m, 2H),
2.13-2.04 (m, 1H), 1.80-1.60 (m, 3H).
Example 4
2-(Pyridin-3-yl)-N-(5-(1-(6-(2-(3-(trifluoromethoxy)phenyl)acetamido)pyrid-
in-3-yl)piperidin-4-yl)-1,3,4-thiadiazol-2-yl)acetamide
[0321] Intermediate 5 (70 mg, 0.15 mmol), 3-Pyridylacetic acid
hydrochloride (31 mg, 0.18 mmol), HATU (67 mg, 0.18 mmol),
N-Ethyldiisopropyl amine (0.1 ml, 0.45 mmol) were dissolved in DMF
(2 ml). This mixture was stirred at rt for 16 h. Reaction mass was
diluted with water and extracted with DCM. Organic layer was washed
with water. Organic layer was dried on anhydrous Na.sub.2SO.sub.4.
DCM was removed on rotavapour to obtain crude. Crude was purified
by column chromatography on 60-120 mesh silica gel using Methanol
and DCM (5:95) as eluent to afford the titled compound (15 mg) as a
pale-brown solid. M.P.: 208-210.degree. C. .sup.1H-NMR (.delta.
ppm, DMSO-d.sub.6, 400 MHz): 12.70 (s, 1H), 10.47 (s, 1H), 8.58 (s,
1H), 8.54 (d, J 4.8, 1H), 8.03 (d, J 2.7, 1H), 7.91-7.85 (m, 2H),
7.51-7.39 (m, 3H), 7.38-7.32 (m, 2H), 7.22 (d, J 7.6, 1H), 3.92 (s,
2H), 3.73 (s, 2H), 3.72-3.65 (m, 2H), 3.31-3.20 (m, 1H), 2.85 (t, J
10.8, 2H), 2.10 (d, J 11.9, 2H), 1.90-1.76 (m, 2H).
Example 5
2-(3-Cyanophenyl)-N-(5-(1-(6-(2-(3-(trifluoromethoxy)phenyl)acetamido)pyri-
din-3-yl)piperidin-4-yl)-1,3,4-thiadiazol-2-yl)acetamide
[0322] Intermediate 5 (100 mg, 0.15 mmol), 3-Cyanophenylacetic acid
(58 mg, 0.36 mmol), HATU (96 mg, 0.25 mmol), N-Ethyldiisopropyl
amine (0.11 ml, 0.63 mnol) were dissolved in DMF (2 ml). This
mixture was stirred at rt for 16 h. Reaction mass was diluted with
water and extracted with DCM. Organic layer was washed with water.
Organic layer was dried on anhydrous Na.sub.2SO.sub.4. DCM was
removed on rotavapour to obtain crude. Crude was purified by
combi-flash using Methanol and DCM (5:95) as eluent to afford the
titled compound (50 mg) as a pale-yellow solid. M.P.:
211-213.degree. C. .sup.1H-NMR (.delta. ppm, DMSO-d.sub.6, 400
MHz): 12.71 (s, 1H), 10.5 (s, 1H), 8.03 (d, J 2.4, 1H), 7.88 (d, J
9, 1H), 7.79-7.73 (m, 2H), 7.65 (d, J 7.9, 1H), 7.57-7.51 (m, 1H),
7.48-7.37 (m, 2H), 7.36-7.31 (m, 2H), 7.23 (d, J 7.9, 1H), 3.90 (s,
2H), 3.75-3.65 (m, 4H), 3.30-3.20 (m, 1H), 2.83 (t, J 11.6, 2H),
2.10 (d, J 12.6, 2H), 1.88-1.78 (m, 2H).
Example 6
2-(Pyridin-2-yl)-N-(5-(4-(5-(2-(3-(trifluoromethoxy)phenyl)acetamido)-1,3,-
4-thiadiazol-2-yl)piperidin-1-yl)pyridin-2-yl)acetamide
[0323] Intermediate 15 (100 mg, 0.25 mmol),
3-(Trifluoromethoxy)phenylacetic acid (66 mg, 0.3 mmol), HATU (114
mg, 0.3 mmol), N-Ethyldiisopropyl amine (0.13 ml, 0.76 mmol) were
dissolved in DMF (1.5 ml). This mixture was stirred at rt for 16 h.
Reaction mass was diluted with water and extracted with DCM.
Organic layer was washed with water. Organic layer was dried on
anhydrous Na.sub.2SO.sub.4. DCM was removed on rotavapour to obtain
crude. Crude was purified by combi-flash using Methanol and DCM
(8:92) as eluent to afford the titled compound (60 mg) as a
pale-yellow solid. M.P.: 188-191.degree. C. .sup.1H-NMR (5 ppm,
DMSO-d.sub.6, 400 MHz): 12.65 (s, 1H), 10.40 (s, 1H), 8.49 (d, J
4.8, 1H), 8.03 (d, J 2.8, 1H), 7.89 (d, J 9, 1H), 7.73 (dt, J 1.8,
7.7, 1H), 7.49-7.43 (m, 1H), 7.42-7.31 (m, 4H), 7.28-7.22 (m, 2H),
3.88 (s, 2H), 3.87 (s, 2H), 3.70 (d, J 12.6, 2H), 3.25-3.20 (m,
1H), 2.84 (t, J 11.6, 2H), 2.11 (d, J 11.5, 2H), 1.90-1.77 (m,
2H).
Example 7
2-(Pyridin-2-yl)-N-(5-(3-(5-(2-(3-(trifluoromethoxy)phenyl)acetamido)-1,3,-
4-thiadiazol-2-yl)piperidin-1-yl)pyridin-2-yl)acetamide
[0324] Intermediate 20 (100 mg, 0.25 mmol),
3-(Trifluoromethoxy)phenylacetic acid (66 mg, 0.3 mmol), HATU (114
mg, 0.3 mmol), N-Ethyldiisopropyl amine (0.13 ml, 0.76 mmol) were
dissolved in DMF (1.5 ml). This mixture was stirred at rt for 16 h.
Reaction mass was diluted with water and extracted with DCM.
Organic layer was washed with water. Organic layer was dried on
anhydrous Na.sub.2SO.sub.4. DCM was removed on rotavapour to obtain
crude. Crude was purified by combi-flash using Methanol and DCM
(5:95) as eluent to afford the titled compound (40 mg) as a
pale-yellow solid. M.P.: 151-153.degree. C. .sup.1H-NMR (.delta.
ppm, DMSO-d.sub.6, 400 MHz): 12.69 (s, 1H), 10.47 (s, 1H), 8.48 (d,
J 4.0, 1H), 8.04 (d, J 2.8, 1H), 7.90 (d, J 9, 1H), 7.73 (dt, J
1.7, 7.7, 1H), 7.49-7.40 (m, 2H), 7.39-7.32 (m, 3H), 7.28-7.23 (m,
2H), 3.87 (s, 4H), 3.70 (d, J 12.4, 1H), 3.49-3.39 (m, 2H),
3.14-3.06 (m, 1H), 2.99-2.90 (m, 1H), 2.10-2.04 (m, 1H), 1.80-1.65
(m, 3H). MS (m/z): 597.8 [M+H].sup.+.
Example 8
2-(pyridin-2-yl)-N-(5-(1-(6-(2-(3-(trifluoromethoxy)phenyl)acetamido)pyrid-
azin-3-yl)piperidin-4-yl)-1,3,4-thiadiazol-2-yl)acetamide
[0325] Intermediate 24 (100 mg, 0.2 mmol), 2-Pyridiylacetic acid
hydrochloride (44 mg, 0.25 mmol), HATU (170 mg, 0.44 mmol),
N-Ethyldiisopropyl amine (0.1 ml, 0.57 mmol) were dissolved in DMF
(2 ml). This mixture was stirred at rt for 1 h. Reaction mass was
diluted with water and extracted with DCM. Organic layer was washed
with water. Organic layer was dried on anhydrous Na.sub.2SO.sub.4.
DCM was removed on rotavapour to obtain crude. Crude was purified
on Combi-flash using DCM and MeOH (95:5) as eluent to afford the
titled compound (60 mg) as a yellow solid. M.P.: 202-205.degree. C.
.sup.1H-NMR (.delta. ppm, DMSO-d.sub.6, 400 MHz): 12.68 (bs, 1H),
10.93 (s, 1H), 8.48 (d, J 4.2, 1H), 7.99 (d, J 9.8, 1H), 7.75 (dt,
J 1.7, 7.7, 1H), 7.47-7.43 (m, 1H), 7.39-7.32 (m, 4H), 7.29-7.21
(m, 2H), 4.30 (d, J 13.2, 2H), 3.99 (s, 2H), 3.78 (s, 2H),
3.40-3.30 (m, 1H), 3.05 (t, J 11.7, 2H), 2.09 (d, J 10.6, 2H),
1.80-1.65 (m, 2H). MS (m/z): 599.6 [M+H].sup.+.
Example 9
2-(Pyridin-2-yl)-N-(5-(1-(6-(2-(3-(trifluoromethoxy)phenyl)acetamido)pyrid-
azin-3-yl)piperidin-3-yl)-1,3,4-thiadiazol-2-yl)acetamide
[0326] Intermediate 28 (80 mg, 0.17 mmol), 2-Pyridiylacetic acid
hydrochloride (34 mg, 0.2 mmol), HATU (138 mg, 0.37 mmol),
N-Ethyldiisopropyl amine (0.08 ml, 0.5 mmol) were dissolved in DMF
(1 ml). This mixture was stirred at rt for 1 h. Reaction mass was
diluted with water and extracted with DCM:MeOH (9:1). Organic layer
was washed with water. Organic layer was dried on anhydrous
Na.sub.2SO.sub.4. DCM and MeOH were removed on rotavapour to obtain
crude. Crude was purified on Combi-flash using DCM and MeOH (95:5)
as eluent to afford the titled compound (20 mg) as a pale-yellow
solid. M.P.: 202-205.degree. C. M.P.: 243-246.degree. C.
.sup.1H-NMR (.delta. ppm, DMSO-d.sub.6, 400 MHz): 12.71 (s, 1H),
10.95 (s, 1H), 8.47 (d, J 4.4, 1H), 7.99 (d, J 9.5, 1H), 7.75 (t, J
7.4, 1H), 7.48-7.32 (m, 5H), 7.30-7.21 (m, 2H), 4.42 (d, J 10, 1H),
4.05-3.97 (m, 3H), 3.78 (s, 2H), 3.45-3.35 (m, 2H), 3.15 (t, J
11.1, 1H), 2.19-2.10 (m, 1H), 1.90-1.74 (m, 2H), 1.67-1.58 (m, 1H).
MS (m/z): 599.5 [M+H].sup.+.
Example 10
2-(Pyridin-3-yl)-N-(5-(1-(6-(2-(3-(trifluoromethoxy)phenyl)acetamido)pyrid-
azin-3-yl)piperidin-4-yl)-1,3,4-thiadiazol-2-yl)acetamide
[0327] Intermediate 24 (80 mg, 0.17 mmol), 3-Pyridiylacetic acid
hydrochloride (35 mg, 0.2 mmol), HATU (140 mg, 0.37 mmol),
N-Ethyldiisopropyl amine (0.08 ml, 0.5 mmol) were dissolved in DMF
(2 ml). This mixture was stirred at rt for 1 h. Reaction mass was
diluted with water to obtain a solid. Solid was filtered and
purified on Combi-flash using DCM and MeOH (94:6) as eluent to
afford the titled compound (25 mg) as a pale-yellow solid. M.P.:
222-223.degree. C. .sup.1H-NMR (.delta. ppm, DMSO-d.sub.6, 400
MHz): 12.72 (s, 1H), 10.93 (s, 1H), 8.50 (bs, 1H), 8.46 (bs, 1H),
7.99 (d, J 9.7, 1H), 7.72 (d, J 7.8, 1H), 7.48-7.42 (m, 1H),
7.39-7.32 (m, 4H), 7.24 (d, J 8.1, 1H), 4.30 (d, J 13.3, 2H), 3.85
(s, 2H), 3.78 (s, 2H), 3.40-3.35 (m, 1H), 3.05 (t, J 11.7, 2H),
2.09 (d, J 11.2, 2H), 1.80-1.67 (m, 2H).
Example 11
2-(3-(Methylsulfonamido)phenyl)-N-(5-(1-(6-(2-(3-(trifluoromethoxy)phenyl)-
acetamido)
pyridazin-3-yl)piperidin-4-yl)-1,3,4-thiadiazol-2-yl)acetamide
[0328] Intermediate 24 (80 mg, 0.17 mmol),
2-(3-(Methylsulfonamido)phenyl)acetic acid (46 mg, 0.2 mmol), HATU
(140 mg, 0.37 mmol), N-Ethyldiisopropyl amine (0.08 ml, 0.5 mmol)
were dissolved in DMF (2 ml). This mixture was stirred at rt for 1
h. Reaction mass was diluted with water. Aqueous layer was
extracted with DCM and MeOH (9:1) mixture. Organic layer was washed
with water and aq. NaHCO.sub.3 solution. Organic layer dried on
anhydrous Na.sub.2SO.sub.4 and organic layer distilled on
rotavapour to obtain crude. Crude was purified on Combi-flash using
DCM and MeOH (95:5) as eluent to afford the titled compound (20 mg)
as a pale-brown solid. M.P.: 228-231.degree. C. .sup.1H-NMR
(.delta. ppm, DMSO-d.sub.6, 400 MHz): 12.69 (s, 1H), 10.93 (s, 1H),
9.72 (s, 1H), 7.99 (d, J 9.8, 1H), 7.49-7.42 (m, 1H), 7.39-7.32 (m,
3H), 7.30-7.22 (m, 2H), 7.16 (s, 1H), 7.10 (d, J 7.9, 1H), 7.04 (d,
J 7.5, 1H), 4.30 (d, J 13, 2H), 3.78 (s, 2H), 3.76 (s, 2H),
3.40-3.30 (m, 1H), 3.05 (t, J 11.6, 2H), 2.97 (s, 3H), 2.13-2.05
(m, 2H), 1.80-1.65 (m, 2H).
Example 12
2-(2-Chlorophenyl)-N-(6-(4-(5-(2-(pyridin-2-yl)acetamido)-1,3,4-thiadiazol-
-2-yl) piperidin-1-yl)pyridazin-3-yl)acetamide
[0329] Intermediate 31 (38 mg, 0.09 mmol), 2-Pyridiylacetic acid
hydrochloride (18 mg, 0.1 mmol), HATU (73 mg, 0.19 mmol),
N-Ethyldiisopropyl amine (0.05 ml, 0.26 mmol) were dissolved in DMF
(1 ml). This mixture was stirred at rt for 1 h. Reaction mass was
diluted with water and extracted with DCM:MeOH (9:1). Organic layer
was washed with water. Organic layer was dried on anhydrous
Na.sub.2SO.sub.4. DCM and MeOH were removed on rotavapour to obtain
crude. Crude was purified on Combi-flash using DCM and MeOH (95:5)
as eluent to afford the titled compound (8 mg) as a pale-brown
solid. M.P.: 225-227.degree. C. .sup.1H-NMR (.delta. ppm,
DMSO-d.sub.6, 400 MHz): 12.68 (s, 1H), 10.92 (s, 1H), 8.48 (d, J
4.1, 1H), 7.98 (d, J 9.8, 1H), 7.75 (dt, J 1.8, 7.7, 1H), 7.44-7.36
(m, 4H), 7.33-7.25 (m, 3H), 4.31 (d, J 13, 2H), 3.99 (s, 2H), 3.90
(s, 2H), 3.40-3.35 (m, 1H), 3.05 (t, J 11.8, 2H), 2.10 (d, J 11.8,
2H), 1.81-1.69 (m, 2H).
Example 13
2-(2-Chlorophenyl)-N-(5-(1-(6-(2-(3-(trifluoromethoxy)phenyl)acetamido)pyr-
idazin-3-yl)piperidin-3-yl)-1,3,4-thiadiazol-2-yl)acetamide
[0330] Intermediate 28 (100 mg, 0.2 mmol), 2-Chlorophenylacetic
acid (42 mg, 0.25 mmol), HATU (173 mg, 0.46 mmol),
N-Ethyldiisopropyl amine (0.1 ml, 0.6 mmol) were dissolved in DMF
(1 ml). This mixture was stirred at rt for 1 h. Reaction mass was
diluted with water and extracted with DCM:MeOH (9:1). Organic layer
was washed with water. Organic layer was dried on anhydrous
Na.sub.2SO.sub.4. DCM and MeOH were removed on rotavapour to obtain
crude. Crude was purified on Combi-flash using DCM and MeOH (97:3)
as eluent to afford the titled compound (40 mg) as a pale-brown
solid. M.P.: 202-205.degree. C. M.P.: 127-130.degree. C.
.sup.1H-NMR (.delta. ppm, DMSO-d.sub.6, 400 MHz): 12.75 (s, 1H),
10.94 (s, 1H), 7.98 (d, J 9.7, 1H), 7.48-7.29 (m, 8H), 7.24 (d, J
7.5, 1H), 4.41 (d, J 10.1, 1H), 4.05-3.95 (m, 3H), 3.78 (s, 2H),
3.40-3.35 (m, 2H), 3.20-3.10 (m, 1H), 2.19-2.10 (m, 1H), 1.88-1.71
(m, 2H), 1.68-1.57 (m, 1H).
Example 14
2-(2-Fluorophenyl)-N-(6-(4-(5-(2-(pyridin-2-yl)acetamido)-1,3,4-thiadiazol-
-2-yl)piperidin-1-yl)pyridazin-3-yl)acetamide
[0331] Intermediate 34 (60 mg, 0.15 mmol), 2-Pyridylacetic acid
hydrochloride (30 mg, 0.17 mmol), HATU (121 mg, 0.32 mmol),
N-Ethyldiisopropyl amine (0.08 ml, 0.5 mmol) were dissolved in DMF
(1 ml). This mixture was stirred at rt for 1 h. Reaction mass was
diluted with water. Aqueous layer was extracted with DCM and MeOH
(9:1) mixture. Organic layer was washed with water and aq.
NaHCO.sub.3 solution. Organic layer dried on anhydrous
Na.sub.2SO.sub.4 and organic layer distilled on rotavapour to
obtain crude. Crude was purified on Combi-flash using DCM and MeOH
(96:4) as eluent to afford the titled compound (13 mg) as a brown
solid. M.P.: 229-231.degree. C. .sup.1H-NMR (.delta. ppm,
DMSO-d.sub.6, 400 MHz): 12.69 (s, 1H), 10.92 (s, 1H), 8.48 (d, J,
1H), 7.98 (d, J 9.9, 1H), 7.75 (dt, J 1.8, 7.7, 1H), 7.40-7.25 (m,
5H), 7.19-7.11 (m, 2H), 4.31 (d, J 13.3, 2H), 3.99 (s, 2H), 3.80
(s, 2H), 3.41-3.35 (m, 1H), 3.05 (t, J 11.4, 2H), 2.10 (d, J 11,
2H), 1.80-1.69 (m, 2H).
Example 15
2-(Pyrazin-2-yl)-N-(5-(1-(6-(2-(3-(trifluoromethoxy)phenyl)acetamido)pyrid-
azin-3-yl)piperidin-4-yl)-1,3,4-thiadiazol-2-yl)acetamide
[0332] Intermediate 24 (80 mg, 0.17 mmol), 2-(pyrazin-2-yl)acetic
acid (27 mg, 0.2 mmol), HATU (140 mg, 0.37 mmol),
N-Ethyldiisopropyl amine (0.08 ml, 0.5 mmol) were dissolved in DMF
(1 ml). This mixture was stirred at rt for 1 h. Reaction mass was
diluted with water. Aqueous layer was extracted with DCM and MeOH
(9:1) mixture. Organic layer was washed with water and aq.
NaHCO.sub.3 solution. Organic layer dried on anhydrous
Na.sub.2SO.sub.4 and organic layer distilled on rotavapour to
obtain crude. Crude was purified on Combi-flash using DCM and MeOH
(94:6) as eluent to afford the titled compound (10 mg) as a
pale-yellow solid. M.P.: 216-218.degree. C. .sup.1H-NMR (.delta.
ppm, DMSO-d.sub.6, 400 MHz): 12.76 (s, 1H), 10.94 (s, 1H), 8.66 (s,
1H), 8.58-8.53 (m, 2H), 7.99 (d, J 9.6, 1H), 7.48-7.42 (m, 1H),
7.39-7.32 (m, 3H), 7.24 (d, J 8.3, 1H), 4.30 (d, J 12.8, 2H), 4.08
(s, 2H), 3.78 (s, 2H), 3.40-3.35 (m, 1H), 3.05 (t, J 11.9, 2H),
2.10 (d, J 13.4, 2H), 1.80-1.65 (m, 2H).
Example 16
2-(Pyridin-2-yl)-N-(5-(1-(6-(2-(3-(trifluoromethoxy)phenyl)acetamido)pyrid-
azin-3-yl)piperidin-4-yl)-1,3,4-thiadiazol-2-yl)acetamide
dihydrochloride
[0333] Example 8 (50 mg, 0.08 mmol) dissolved in THF (15 ml) and
added Et.sub.2O.HCl (5 ml). This mixture was stirred under nitrogen
atmosphere for 30 mins. After 30 mins, removed THF and diethyl
ether on rotavapour to obtain a residue. Residue was triturated
with diethyl ether to obtain the titled compound (40 mg) as a brown
solid. M.P.: 240-243.degree. C. .sup.1H-NMR (.delta. ppm,
DMSO-d.sub.6, 400 MHz): 12.95 (bs, 1H), 11.45 (s, 1H), 8.76 (d, J
5, 1H), 8.30 (t, J 6.6, 1H), 8.24 (d, J 10.1, 1H), 7.94 (d, J 10,
1H), 7.83 (d, J 7.7, 1H), 7.76 (t, J 6.4, 1H), 7.48-7.42 (m, 1H),
7.38-7.34 (m, 2H), 7.25 (d, J 8, 1H), 4.35-4.27 (m, 4H), 3.84 (s,
2H), 3.51-3.44 (m, 1H), 3.39-3.29 (m, 2H), 2.20-2.10 (m, 2H),
1.91-1.79 (m, 2H).
Example 17
2-(Pyridin-2-yl)-N-(6-(4-(5-(2-(3-(trifluoromethoxy)phenyl)acetamido)-1,3,-
4-thiadiazol-2-yl)piperidin-1-yl)pyridazin-3-yl)acetamide
[0334] Intermediate 36 (150 mg, 0.38 mmol),
3-(Trifluoromethoxy)phenylacetic acid (100 mg, 0.45 mmol), HATU
(316 mg, 0.37 mmol), N-Ethyldiisopropyl amine (0.2 ml, 1.134 mmol)
were dissolved in DMF (1 ml). This mixture was stirred at rt for 1
h. Reaction mass was poured in to water to obtain a solid. Solid
was filtered and purified the solid by column chromatography on
60-120 silica gel using MeOH and DCM (8:92) as eluent to afford the
titled compound (30 mg) as a pale-brown solid. M.P.:
220-222.degree. C. .sup.1H-NMR (.delta. ppm, DMSO-d, 400 MHz):
12.70 (s, 1H), 10.92 (s, 1H), 8.49 (s, 1H), 8.01 (d, J 8.7, 1H),
7.80-7.70 (m, 1H), 7.50-7.20 (m, 7H), 4.30 (d, J 12.2, 2H), 3.92
(s, 2H), 3.87 (s, 2H), 3.50-3.40 (m, 1H), 3.05 (t, J 12.2, 2H),
2.10 (d, J 11.1, 2H), 1.80-1.65 (m, 2H).
Example 18
2-(Pyridin-3-yl)-N-(6-(4-(5-(2-(3-(trifluoromethoxy)phenyl)acetamido)-1,3,-
4-thiadiazol-2-yl)piperidin-1-yl)pyridazin-3-yl)acetamide
[0335] Intermediate 38 (75 mg, 0.19 mmol),
3-(Trifluoromethoxy)phenylacetic acid (50 mg, 0.22 mmol), HATU (158
mg, 0.42 mmol), N-Ethyldiisopropyl amine (0.1 ml, 0.5 mmol) were
dissolved in DMF (1 ml). This mixture was stirred at rt for 1 h.
Reaction mass was poured in to water to obtain a solid. Solid was
filtered and purified by column chromatography on 60-120 mesh
silica gel using MeOH and DCM (7:93) as eluent to afford the titled
compound (15 mg) as a pale-brown solid. M.P.: 125-127.degree. C.
.sup.1H-NMR (.delta. ppm, DMSO-d.sub.6, 400 MHz): 12.70 (s, 1H),
10.97 (s, 1H), 8.51 (s, 11), 8.45 (s, 11), 7.99 (d, J 9.5, 1H),
7.73 (d, J 6.9, 1H), 7.50-7.42 (m, 11), 7.40-7.30 (m, 4H), 7.26 (d,
J 7.4, 1H), 4.30 (d, J 12.2, 2H), 3.87 (s, 2H), 3.76 (s, 2H),
3.40-3.35 (m, 1H), 3.04 (t, J 12, 2H), 2.09 (d, J 12, 2H),
1.80-1.68 (m, 2H).
Example 19
2-(Pyridin-3-yl)-N-(6-(4-(5-(2-(2,3,6-trifluorophenyl)acetamido)-1,3,4-thi-
adiazol-2-yl)piperidin-1-yl)pyridazin-3-yl)acetamide
[0336] Intermediate 38 (75 mg, 0.19 mmol),
2,3,6-Trifluorophenylacetic acid (120 mg, 0.30 mnol), HATU (255 mg,
0.67 mmol), N-Ethyldiisopropyl amine (0.15 ml, 0.92 mmol) were
dissolved in DMF (1 ml). This mixture was stirred at rt for 1 h.
Reaction mass was poured in to water to obtain a solid. Solid was
filtered and dried on high vacuum. This solid was triturated with
Et.sub.2O to obtain the titled compound (70 mg) as a brown solid.
M.P.: 252-254.degree. C. .sup.1H-NMR (.delta. ppm, DMSO-d.sub.6,
400 MHz): 12.84 (s, 11), 10.96 (s, 11), 8.51 (s, 11), 8.44 (s, 11),
7.98 (d, J 8.1, 1H), 7.76-7.70 (m, 1H), 7.51-7.29 (m, 3H),
7.20-7.10 (m, 1H), 4.30 (d, J 10.6, 2H), 3.96 (s, 2H), 3.76 (s,
2H), 3.41-3.35 (m, 1H), 3.10-3.00 (m, 2H), 2.10 (d, J 10, 2H),
1.81-1.69 (m, 2H).
Example 20
2-(Pyridin-2-yl)-N-(6-(4-(5-(2-(2,3,6-trifluorophenyl)acetamido)-1,3,4-thi-
adiazol-2-yl)piperidin-1-yl)pyridazin-3-yl)acetamide
[0337] Intermediate 36 (100 mg, 0.25 mmol),
2,3,6-Trifluorophenylacetic acid (58 mg, 0.30 mmol), HATU (211 mg,
0.55 mmol), N-Ethyldiisopropyl amine (0.13 ml, 0.76 mmol) were
dissolved in DMF (1 ml). This mixture was stirred at rt for 1 h.
Reaction mass was poured in to water to obtain a solid. Solid was
filtered and purified the solid by column chromatography on 60-120
silica gel using MeOH and DCM (4:96) as eluent to afford the titled
compound (40 mg) as a brown solid. M.P.: 214-218.degree. C.
.sup.1H-NMR (.delta. ppm, DMSO-d.sub.6, 400 MHz): 12.84 (s, 1H),
10.92 (s, 11), 8.49 (d, J 3.7, 1H), 8.01 (d, J 9.8, 1H), 7.74 (dt,
J 1.6, 7.7, 1H), 7.50-7.43 (m, 1H), 7.40-7.35 (m, 2H), 7.28-7.23
(m, 1H), 7.20-7.13 (m, 1H), 4.31 (d, J 13.3, 2H), 3.97 (s, 2H),
3.91 (s, 2H), 3.40-3.30 (m, 1H), 3.05 (t, J 11.8, 2H), 2.10 (d, J
11.2, 2H), 1.80-1.70 (m, 2H).
Example 21
2-(2,3-Difluorophenyl)-N-(5-(1-(6-(2-(pyridin-2-yl)acetamido)pyridazin-3-y-
l)piperidin-4-yl)-1,3,4-thiadiazol-2-yl)acetamide
[0338] Intermediate 36 (100 mg, 0.25 mmol),
2,3-Difluorophenylacetic acid (52 mg, 0.30 mmol), HATU (211 mg,
0.55 mmol), N-Ethyldiisopropyl amine (0.13 ml, 0.76 mmol) were
dissolved in DMF (1 ml). This mixture was stirred at rt for 1 h.
Reaction mass was poured in to water to obtain a solid. Solid was
filtered and purified the solid by column chromatography on 60-120
silica gel using MeOH and DCM (4:96) as eluent to afford the titled
compound (18 mg) as a brown solid. M.P.: 203-206.degree. C.
.sup.1H-NMR (.delta. ppm, DMSO-d.sub.6, 400 MHz): 12.76 (s, 1H),
10.92 (s, 1H), 8.49 (d, J 3.7, 1H), 8.01 (d, J 10.1, 1H), 7.75 (t,
J 6.4, 1H), 7.40-7.33 (m, 3H), 7.27-7.23 (m, 1H), 7.20-7.15 (m,
2H), 4.30 (d, J 13.2, 2H), 3.95 (s, 2H), 3.91 (s, 2H), 3.40-3.35
(m, 1H), 3.05 (t, J 12.4, 2H), 2.10 (d, J 11.4, 2H), 1.80-1.70 (m,
2H).
Example 22
2-(3,4-Difluorophenyl)-N-(5-(1-(6-(2-(pyridin-2-yl)acetamido)pyridazin-3-y-
l)piperidin-4-yl)-1,3,4-thiadiazol-2-yl)acetamide
[0339] Intermediate 36 (100 mg, 0.25 mmol),
3,4-Difluorophenylacetic acid (52 mg, 0.30 mmol), HATU (211 mg,
0.55 mmol), N-Ethyldiisopropyl amine (0.13 ml, 0.76 mmol) were
dissolved in DMF (1 ml). This mixture was stirred at rt for 1 h.
Reaction mass was poured in to water to obtain a solid. Solid was
filtered and purified the solid by column chromatography on 60-120
silica gel using MeOH and DCM (4:96) as eluent to afford the titled
compound (37 mg) as a pale-brown solid. M.P.: 211-214.degree. C.
.sup.1H-NMR (.delta. ppm, DMSO-d.sub.6, 400 MHz): 12.67 (s, 1H),
10.92 (s, 1H), 8.49 (d, J 4, 1H), 8.01 (d, J 9.8, 1H), 7.74 (dt, J
1.7, 7.7, 1H), 7.41-7.33 (m, 4H), 7.29-7.23 (m, 1H), 7.16-7.10 (m,
1H), 4.30 (d, J 13.1, 2H), 3.91 (s, 2H), 3.81 (s, 2H), 3.40-3.35
(m, 1H), 3.05 (t, J 11.4, 22H), 2.09 (d, J 11.2, 2H), 1.80-1.58 (m,
2H).
Example 23
2-(2-Fluorophenyl)-N-(5-(1-(6-(2-(pyridin-2-yl)acetamido)pyridazin-3-yl)pi-
peridin-4-yl)-1,3,4-thiadiazol-2-yl)acetamide
[0340] Intermediate 36 (100 mg, 0.25 mmol), 2-Fluorophenylacetic
acid (47 mg, 0.30 mmol), HATU (211 mg, 0.55 mmol),
N-Ethyldiisopropyl amine (0.13 ml, 0.76 mmol) were dissolved in DMF
(1 ml). This mixture was stirred at rt for 1 h. Reaction mass was
poured in to water to obtain a solid. Solid was filtered and
purified the solid by column chromatography on 60-120 silica gel
using MeOH and DCM (4:96) as eluent to afford the titled compound
(40 mg) as a pale-brown solid. M.P.: 220-223.degree. C. .sup.1H-NMR
(.delta. ppm, DMSO-d.sub.6, 400 MHz): 12.71 (bs, 1H), 10.92 (s,
1H), 8.49 (d, J 4.1, 1H), 8.01 (d, J 9.5, 1H), 7.74 (t, J 7.8, 1H),
7.41-7.30 (m, 4H), 7.29-7.23 (m, 1H), 7.20-7.13 (m, 2H), 4.30 (d, J
13, 2H), 3.91 (s, 2H), 3.87 (s, 2H), 3.41-3.35 (m, 1H), 3.05 (t, J
11.6, 2H), 2.10 (d, J 11.1, 2H), 1.81-1.67 (m, 2H).
Example 24
2-(3-Fluorophenyl)-N-(5-(1-(6-(2-(pyridin-2-yl)acetamido)pyridazin-3-yl)pi-
peridin-4-yl)-1,3,4-thiadiazol-2-yl)acetamide
[0341] Intermediate 36 (100 mg, 0.25 mmol), 3-Fluorophenylacetic
acid (47 mg, 0.30 mmol), HATU (211 mg, 0.55 mmol),
N-Ethyldiisopropyl amine (0.13 ml, 0.76 mmol) were dissolved in DMF
(1 ml). This mixture was stirred at rt for 1 h. Reaction mass was
poured in to water to obtain a solid. Solid was filtered and
purified the solid by column chromatography on 60-120 silica gel
using MeOH and DCM (7:93) as eluent to afford the titled compound
(40 mg) as a brown solid. M.P.: 210-213.degree. C. .sup.1H-NMR
(.delta. ppm, DMSO-d.sub.6, 400 MHz): 12.69 (bs, 1H), 10.92 (s,
1H), 8.49 (d, J 4, 1H), 8.01 (d, J 9.8, 1H), 7.74 (dt, J 1.7, 7.4,
1H), 7.40-7.33 (m, 3H), 7.27-7.23 (m, 1H), 7.18-7.05 (m, 3H), 4.30
(d, J 13.1, 2H), 3.91 (s, 2H), 3.82 (s, 2H), 3.40-3.35 (m, 1H),
3.05 (t, J 11.8, 2H), 2.09 (d, J 11.4, 2H), 1.80-1.67 (m, 2H).
Example 25
2-(4-Fluorophenyl)-N-(5-(1-(6-(2-(pyridin-2-yl)acetamido)pyridazin-3-yl)pi-
peridin-4-yl)-1,3,4-thiadiazol-2-yl)acetamide
[0342] Intermediate 36 (100 mg, 0.25 mmol), 4-Fluorophenylacetic
acid (47 mg, 0.30 mmol), HATU (211 mg, 0.55 mmol),
N-Ethyldiisopropyl amine (0.13 ml, 0.76 mmol) were dissolved in DMF
(1 ml). This mixture was stirred at rt for 1 h. Reaction mass was
poured in to water to obtain a solid. Solid was filtered and
purified the solid by column chromatography on 60-120 silica gel
using MeOH and DCM (4:96) as eluent to afford the titled compound
(25 mg) as a brown solid. M.P.: 194-197.degree. C. .sup.1H-NMR
(.delta. ppm, DMSO-d.sub.6, 400 MHz): 12.67 (s, 1H), 10.92 (s, 1H),
8.49 (d, J 4.6, 1H), 8.01 (d, J 9.8, 1H), 7.74 (dt, J 1.7, 7.7,
1H), 7.40-7.31 (m, 4H), 7.28-7.22 (m, 1H), 7.17-7.11 (m, 2H), 4.30
(d, J 13.1, 2H), 3.91 (s, 2H), 3.78 (s, 2H), 3.40-3.35 (m, 1H),
3.05 (t, J 11.8, 2H), 2.09 (d, J 11.6, 2H), 1.80-1.65 (m, 2H).
Example 26
2-(2-Methoxyphenyl)-N-(5-(1-(6-(2-(pyridin-2-yl)acetamido)pyridazin-3-yl)p-
iperidin-4-yl)-1,3,4-thiadiazol-2-yl)acetamide
[0343] Intermediate 36 (100 mg, 0.25 mmol), 2-Methoxyphenylacetic
acid (50 mg, 0.30 mmol), HATU (211 mg, 0.55 mmol),
N-Ethyldiisopropyl amine (0.13 ml, 0.76 mmol) were dissolved in DMF
(1 ml). This mixture was stirred at rt for 1 h. Reaction mass was
poured in to water to obtain a solid. Solid was filtered and
purified the solid by column chromatography on 60-120 silica gel
using MeOH and DCM (4:96) as eluent to afford the titled compound
(37 mg) as a yellow solid. M.P.: 160-163.degree. C. .sup.1H-NMR
(.delta. ppm, DMSO-d.sub.6, 400 MHz): 12.55 (s, 1H), 10.92 (s, 1H),
8.49 (d, J 4.1, 1H), 8.01 (d, J 9.8, 1H), 7.75 (dt, J 1.8, 7.7,
1H), 7.40-7.36 (m, 2H), 7.29-7.23 (m, 2H), 7.21-7.18 (m, 1H), 6.96
(d, J 8.2, 1H), 6.91-6.86 (m, 1H), 4.31 (d, J 13.2, 2H), 3.91 (s,
2H), 3.76 (s, 2H), 3.71 (s, 3H), 3.40-3.33 (m, 1H), 3.05 (t, J
11.6, 2H), 2.10 (d, J 11.2, 2H), 1.80-1.69 (m, 2H).
Example 27
2-(2-Chlorophenyl)-N-(5-(1-(6-(2-(pyridin-2-yl)acetamido)pyridazin-3-yl)pi-
peridin-4-yl)-1,3,4-thiadiazol-2-yl)acetamide
[0344] Intermediate 36 (100 mg, 0.25 mmol), 2-Chlorophenylacetic
acid (52 mg, 0.30 mmol), HATU (211 mg, 0.55 mmol),
N-Ethyldiisopropyl amine (0.13 ml, 0.76 mmol) were dissolved in DMF
(1 ml). This mixture was stirred at rt for 1 h. Reaction mass was
poured in to water to obtain a solid. Solid was filtered and
purified the solid by column chromatography on 60-120 silica gel
using MeOH and DCM (4:96) as eluent to afford the titled compound
(35 mg) as a brown solid. M.P.: 194-196.degree. C. .sup.1H-NMR
(.delta. ppm, DMSO-d.sub.6, 400 MHz): 12.73 (s, 1H), 10.91 (s, 1H),
8.49 (d, J 3.9, 1H), 8.01 (d, J 8.6, 1H), 7.77-7.71 (m, 1H),
7.47-7.35 (m, 4H), 7.34-7.28 (m, 2H), 7.27-7.22 (m, 1H), 4.30 (d, J
13.1, 2H), 3.98 (s, 2H), 3.91 (s, 2H), 3.41-3.36 (m, 1H), 3.05 (t,
J 11.8, 2H), 2.10 (d, J 11.5, 2H), 1.80-1.66 (m, 2H).
Example 28
2-(5-Chloro-2-(trifluoromethyl)phenyl)-N-(5-(1-(6-(2-(pyridin-2-yl)acetami-
do)
pyridazin-3-yl)piperidin-4-yl)-1,3,4-thiadiazol-2-yl)acetamide
[0345] Intermediate 36 (100 mg, 0.25 mmol),
5-Chloro-2-trifluoromethylphenylacetic acid (72 mg, 0.30 mmol),
HATU (211 mg, 0.55 mmol), N-Ethyldiisopropyl amine (0.13 ml, 0.76
mmol) were dissolved in DMF (1 ml). This mixture was stirred at rt
for 1 h. Reaction mass was poured in to water to obtain a solid.
Solid was filtered and purified the solid by column chromatography
on 60-120 silica gel using MeOH and DCM (4:96) as eluent to afford
the titled compound (17 mg) as a yellow solid. M.P.:
232-234.degree. C. .sup.1H-NMR (.delta. ppm, DMSO-d.sub.6, 400
MHz): 12.74 (s, 1H), 10.91 (s, 1H), 8.49 (d, J 4.5, 1H), 8.01 (d, J
9.6, 1H), 7.77-7.72 (m, 2H), 7.67 (s, 1H), 7.60 (d, J 8.8, 1H),
7.41-7.34 (m, 2H), 7.27-7.23 (m, 1H), 4.30 (d, J 13, 2H), 4.09 (s,
2H), 3.91 (s, 2H), 3.41-3.30 (m, 1H), 3.05 (t, J 12, 2H), 2.10 (d,
J 11.2, 2H), 1.80-1.67 (m, 2H).
Example 29
2-(4-Chlorophenyl)-N-(5-(1-(6-(2-(pyridin-2-yl)acetamido)pyridazin-3-yl)pi-
peridin-4-yl)-1,3,4-thiadiazol-2-yl)acetamide
[0346] Intermediate 36 (100 mg, 0.25 mmol), 4-Chlorophenylacetic
acid (52 mg, 0.30 mmol), HATU (211 mg, 0.55 mmol),
N-Ethyldiisopropyl amine (0.13 ml, 0.76 mmol) were dissolved in DMF
(1 ml). This mixture was stirred at rt for 1 h. Reaction mass was
poured in to water to obtain a solid. Solid was filtered and
purified the solid by column chromatography on 60-120 silica gel
using MeOH and DCM (4:96) as eluent to afford the titled compound
(40 mg) as a pale-yellow solid. M.P.: 220-222.degree. C.
.sup.1H-NMR (.delta. ppm, DMSO-d.sub.6, 400 MHz): 12.68 (s, 1H),
10.91 (s, 1H), 8.49 (d, J 3.5, 1H), 8.01 (d, J 9.7, 1H), 7.75 (dt,
J 1.5, 7.7, 1H), 7.41-7.32 (m, 611), 7.28-7.23 (m, 1H), 4.30 (d, J
13.3, 2H), 3.91 (s, 2H), 3.79 (s, 2H), 3.41-3.35 (m, 1H), 3.05 (t,
J 11.8, 2H), 2.09 (d, J 12.7, 2H), 1.80-1.65 (m, 2H).
Example 30
2-(Quinolin-6-yl)-N-(5-(1-(6-(2-(3-(trifluoromethoxy)phenyl)acetamido)pyri-
dazin-3-yl)piperidin-4-yl)-1,3,4-thiadiazol-2-yl)acetamide
[0347] Intermediate 24 (100 mg, 0.25 mmol), Quinoline-6-acetic acid
(47 mg, 0.25 mmol), HATU (173 mg, 0.46 mmol), N-Ethyldiisopropyl
amine (0.11 ml, 0.62 mmol) were dissolved in DMF (1 ml). This
mixture was stirred at rt for 1 h. Reaction mass was poured in to
water to obtain a solid. Solid was filtered and purified the solid
by column chromatography on 60-120 silica gel using MeOH and DCM
(4:96) as eluent to afford the titled compound (35 mg) as an
off-white solid. M.P.: 219-221.degree. C. .sup.1H-NMR (.delta. ppm,
DMSO-d.sub.6, 400 MHz): 12.77 (s, 1H), 10.92 (s, 1H), 8.87-8.85 (m,
1H), 8.33 (d, J 8.6, 1H), 8.01-7.95 (m, 2H), 7.88 (s, 1H), 7.70
(dd, J 1.7, 8.7, 1H), 7.54-7.49 (m, 1H), 7.48-7.42 (m, 1H),
7.37-7.32 (m, 3H), 7.23 (d, J 8, 1H), 4.29 (d, J 13, 2H), 4.02 (s,
2H), 3.78 (s, 2H), 3.40-3.32 (m, 1H), 3.04 (t, J 11.7, 2H), 2.09
(d, J 12.2, 2H), 1.80-1.67 (m, 2H).
Example 31
2-o-Tolyl-N-(5-(1-(6-(2-(3-(trifluoromethoxy)phenyl)acetamido)pyridazin-3--
yl)piperidin-4-yl)-1,3,4-thiadiazol-2-yl)acetamide
[0348] Intermediate 24 (100 mg, 0.25 mmol), O-Tolylacetic acid (37
mg, 0.25 mmol), HATU (173 mg, 0.46 mmol), N-Ethyldiisopropyl amine
(0.11 ml, 0.62 mmol) were dissolved in DMF (1 ml). This mixture was
stirred at rt for 1 h. Reaction mass was poured in to water to
obtain a solid. Solid was filtered and purified the solid by column
chromatography on 60-120 silica gel using MeOH and DCM (5:95) as
eluent to afford the titled compound (40 mg) as an off-white solid.
M.P.: 198-200.degree. C. .sup.1H-NMR (.delta. ppm, DMSO-d.sub.6,
400 MHz): 12.69 (bs, 1H), 10.94 (s, 1H), 7.98 (d, J 9.8, 1H),
7.48-7.42 (m, 1H), 7.38-7.32 (m, 3H), 7.26-7.19 (m, 2H), 7.17-7.09
(m, 3H), 4.29 (d, J 13.1, 2H), 3.79 (s, 2H), 3.77 (s, 2H),
3.40-3.32 (m, 1H), 3.04 (t, J 11.7, 2H), 2.24 (s, 3H), 2.08 (d, J
10.9, 2H), 1.80-1.67 (m, 2H).
Example 32
N-(6-(4-(5-(2-(1H-indol-3-yl)acetamido)-1,3,4-thiadiazol-2-yl)piperidin-1--
yl)pyridazin-3-yl)-2-(3-(trifluoromethoxy)phenyl)acetamide
[0349] Intermediate 24 (100 mg, 0.25 mmol), Indole-3-acetic acid
(43 mg, 0.25 mmol), HATU (173 mg, 0.46 mmol), N-Ethyldiisopropyl
amine (0.11 ml, 0.62 mmol) were dissolved in DMF (1 ml). This
mixture was stirred at rt for 1 h. Reaction mass was poured in to
water to obtain a solid. Solid was filtered and purified the solid
by column chromatography on 60-120 silica gel using MeOH and DCM
(5:95) as eluent to afford the titled compound (35 mg) as a brown
solid. M.P.: 220-223.degree. C. .sup.1H-NMR (.delta. ppm,
DMSO-d.sub.6, 400 MHz): 12.60 (s, 1H), 10.92 (s, 1H), 7.98 (d, J
9.9, 1H), 7.55 (d, J 7.8, 1H), 7.48-7.42 (m, 1H), 7.38-7.32 (m,
411), 7.28-7.21 (m, 2H), 7.08-7.03 (m, 1H), 6.98-6.94 (m, 1H), 4.29
(d, J 13, 2H), 3.87 (s, 2H), 3.78 (s, 2H), 3.40-3.30 (m, 1H), 3.04
(t, J 11.8, 2H), 1.80-1.66 (m, 2H).
Example 33
2-(2-Fluorophenyl)-N-(6-(4-(5-(2-(pyrazin-2-yl)acetamido)-1,3,4-thiadiazol-
-2-yl)piperidin-1-yl)pyridazin-3-yl)acetamide
[0350] Intermediate 34 (300 mg, 0.75 mmol), 2-(pyrazin-2-yl)acetic
acid (120 mg, 0.87 mmol), HATU (600 mg, 1.59 mmol),
N-Ethyldiisopropyl amine (0.3 ml, 2.17 mmol) were dissolved in DMF
(4 ml). This mixture was stirred at rt for 1 h. Reaction mass was
poured in to water to obtain a solid. Solid was filtered and
purified the solid by column chromatography on 60-120 silica gel
using MeOH and DCM (5:95) as eluent to afford the titled compound
(13 mg) as a Pale-brown solid. M.P.: 232-234.degree. C. .sup.1H-NMR
(.delta. ppm, DMSO-d.sub.6, 400 MHz): 12.74 (s, 1H), 10.90 (s, 1H),
8.65 (s, 1H), 8.57-8.52 (m, 2H), 7.98 (d, J 9.8, 1H), 7.40-7.36 (m,
2H), 7.35-7.25 (m, 1H), 7.18-7.12 (m, 2H), 4.30 (d, J 13.3, 2H),
4.06 (s, 2H), 3.79 (s, 2H), 3.40-3.31 (m, 1H), 3.05 (t, J 12, 2H),
2.10 (d, J 10.7, 2H), 1.80-1.66 (m, 2H).
Example 34
2-(3-(Azetidin-1-yl)phenyl)-N-(5-(1-(6-(2-(3-(trifluoromethoxy)phenyl)acet-
amido)
pyridazin-3-yl)piperidin-4-yl)-1,3,4-thiadiazol-2-yl)acetamide
[0351] Intermediate 24 (100 mg, 0.25 mnol),
2-(3-(azetidin-1-yl)phenyl)acetic acid (45 mg, 0.25 mmol), HATU
(170 mg, 0.44 mmol), N-Ethyldiisopropyl amine (0.11 ml, 0.62 mmol)
were dissolved in DMF (2 ml). This mixture was stirred at rt for 1
h. Reaction mass was poured in to water to obtain a solid. Solid
was filtered and purified the solid by column chromatography on
60-120 silica gel using MeOH and DCM (4:96) as eluent to afford the
titled compound (40 mg) as an Off-White solid. M.P.:
140-142.degree. C. .sup.1H-NMR (.delta. ppm, DMSO-d.sub.6, 400
MHz): 12.60 (s, 1H), 10.93 (s, 1H), 7.99 (d, J 9.8, 1H), 7.48-7.42
(m, 1H), 7.40-7.32 (m, 3H), 7.24 (d, J 7.6, 1H), 7.07 (t, J 7.8,
1H), 6.59 (d, J 7.3, 1H), 6.36 (s, 1H), 6.28 (d, J 7.8, 1H), 4.29
(d, J 12.9, 2H), 3.81-3.73 (m, 6H), 3.66 (s, 2H), 3.40-3.31 (m,
1H), 3.04 (t, J 11.8, 2H), 2.81-2.72 (m, 2H), 2.08 (d, J 11, 2H),
1.80-1.66 (m, 2H).
Example 35
2-(3-Chlorophenyl)-N-(5-(1-(6-(2-(pyridin-2-yl)acetamido)pyridazin-3-yl)pi-
peridin-4-yl)-1,3,4-thiadiazol-2-yl)acetamide
[0352] Intermediate 36 (100 mg, 0.25 mmol), 3-Chlorophenylacetic
acid (52 mg, 0.30 mmol), HATU (211 mg, 0.55 mmol),
N-Ethyldiisopropyl amine (0.13 ml, 0.76 mmol) were dissolved in DMF
(1 ml). This mixture was stirred at rt for 1 h. Reaction mass was
poured in to water to obtain a solid. Solid was filtered and
purified the solid by column chromatography on 60-120 silica gel
using MeOH and DCM (4:96) as eluent to afford the titled compound
(35 mg) as a pale-yellow solid. M.P.: 198-201.degree. C.
.sup.1H-NMR (.delta. ppm, DMSO-d.sub.6, 400 MHz): 12.68 (s, 1H),
10.91 (s, 1H), 8.49 (d, J 4.2, 1H), 8.01 (d, J 9.8, 1H), 7.76-7.72
(m, 1H), 7.40-7.30 (m, 511), 7.28-7.22 (m, 2H), 4.30 (d, J 13.2,
2H), 3.91 (s, 2H), 3.82 (s, 2H), 3.40-3.29 (m, 1H), 3.05 (t, J
11.8, 2H), 2.10 (d, J 11.7, 2H), 1.80-1.77 (m, 2H).
Example 36
3-Hydroxy-2-phenyl-N-(5-(1-(6-(2-(3-(trifluoromethoxy)phenyl)acetamido)pyr-
idazin-3-yl)piperidin-4-yl)-1,3,4-thiadiazol-2-yl)propanamide
[0353] Intermediate 24 (100 mg, 0.21 mmol), Tropic acid (42 mg,
0.25 mmol), HATU (173 mg, 0.46 mmol), N-Ethyldiisopropyl amine
(0.11 ml, 0.62 mmol) were dissolved in DMF (2 ml). This mixture was
stirred at rt for 1 h. Reaction mass was poured in to water to
obtain a solid. Solid was filtered and purified the solid by column
chromatography on 60-120 silica gel using MeOH and DCM (5:95) as
eluent to afford the titled compound (12 mg) as an Off-White solid.
M.P.: 212-214.degree. C. .sup.1H-NMR (.delta. ppm, DMSO-d.sub.6,
400 MHz): 12.66 (s, 1H), 10.94 (s, 1H), 7.99 (d, J 9.9, 1H),
7.48-7.42 (m, 4H), 7.39-7.22 (m, 911), 5.06-5.02 (m, 1H), 4.30 (d,
J 13.8, 2H), 4.10-4.00 (m, 2H), 3.78 (s, 2H), 3.62-3.57 (m, 1H),
3.40-3.30 (m, 2H), 3.05 (t, J 12.4, 2H), 2.09 (d, J 10.8, 2H),
1.80-1.68 (m, 2H).
Example 37
(R)-2-hydroxy-2-phenyl-N-(5-(1-(6-(2-(3-(trifluoromethoxy)phenyl)acetamido-
)pyridazin-3-yl)piperidin-4-yl)-1,3,4-thiadiazol-2-yl)acetamide
[0354] Intermediate 24 (100 mg, 0.21 mmol), (R)-(-)-Mandelic acid
(35 mg, 0.25 mmol), HATU (173 mg, 0.46 mmol), N-Ethyldiisopropyl
amine (0.11 ml, 0.62 mmol) were dissolved in DMF (2 ml). This
mixture was stirred at rt for 1 h. Reaction mass was poured in to
water to obtain a solid. Solid was filtered and purified the solid
by column chromatography on 60-120 silica gel using MeOH and DCM
(5:95) as eluent to afford the titled compound (15 mg) as a
pale-brown solid. .sup.1H-NMR (.delta. ppm, DMSO-d.sub.6, 400 MHz):
12.41 (s, 1H), 10.93 (s, 1H), 7.98 (d, J 9.7, 1H), 7.50-7.30 (m,
10H), 6.32 (bs, 1H), 5.30 (s, 1H), 4.29 (d, J 13, 2H), 3.78 (s,
2H), 3.50-3.40 (m, 1H), 3.04 (t, J 11.7, 2H), 2.09 (d, J 11, 2H),
1.80-1.65 (m, 2H).
Example 38
2-(3-(3-Fluoroazetidin-1-yl)phenyl)-N-(5-(1-(6-(2-(3-(trifluoromethoxy)phe-
nyl)acetamido)pyridazin-3-yl)piperidin-4-yl)-1,3,4-thiadiazol-2-yl)acetami-
de
[0355] Intermediate 24 (100 mg, 0.21 mmol),
2-(3-(3-Fluoroazetidin-1-yl)phenyl)acetic acid (53 mg, 0.25 mmol),
HATU (173 mg, 0.46 mmol), N-Ethyldiisopropyl amine (0.11 ml, 0.62
mmol) were dissolved in DMF (2 ml). This mixture was stirred at rt
for 1 h. Reaction mass was poured in to water to obtain a solid.
Solid was filtered and purified the solid by column chromatography
on 60-120 silica gel using MeOH and DCM (4:96) as eluent to afford
the titled compound (40 mg) as an Off-White solid. .sup.1H-NMR
(.delta. ppm, DMSO-d.sub.6, 400 MHz): 12.62 (s, 1H), 10.93 (s, 1H),
7.99 (d, J 9.7, 1H), 7.48-7.42 (m, 1H), 7.39-7.22 (m, 4H), 7.11 (t,
J 5.8, 1H), 6.65 (d, J 7.5, 1H), 6.43 (s, 1H), 6.36 (d, J 8, 1H),
4.29 (d, J 13.2, 2H), 4.18-4.06 (m, 2H), 3.90-3.76 (m, 5H), 3.68
(s, 2H), 3.40-3.30 (m, 1H), 3.04 (t, J 11.8, 2H), 2.07 (d, J 12.2,
2H), 1.80-1.66 (m, 2H).
Example 39
2-(Pyridin-2-yl)-N-(5-((1-(6-(2-(3-(trifluoromethoxy)phenyl)acetamido)pyri-
dazin-3-yl)piperidin-4-yl)methyl)-1,3,4-thiadiazol-2-yl)acetamide
[0356] Intermediate 41 (34 mg, 0.07 mmol), 2-Pyridylacetic acid
hydrochloride (15 mg, 0.086 unol), HATU (58 mg, 0.15 mmol),
N-Ethyldiisopropyl amine (0.04 ml, 0.15 mmol) were dissolved in DMF
(2 ml). This mixture was stirred at rt for 1 h. Reaction mass was
poured in to water to obtain a solid. Solid was filtered and
purified the solid by combi-flash using MeOH and DCM (4:96) as
eluent to afford the titled compound (2 mg) as an off-white solid.
.sup.1H-NMR (.delta. ppm, DMSO-d.sub.6, 400 MHz): 12.67 (s, 11),
10.90 (s, 11), 8.48 (d, J 4, 1H), 7.95 (d, J 9.8, 1H), 7.76 (dt, J
2, 7.7, 1H), 7.48-7.22 (m, 711), 4.23 (d, J 12.9, 2H), 3.99 (s,
2H), 3.77 (s, 2H), 2.93 (d, J 7, 2H), 2.82 (t, J 12, 2H), 2.00-1.90
(m, 1H), 1.72 (d, J 12, 2H), 1.30-1.18 (m, 2H).
Biological Assays
[0357] The pharmacological properties of the compounds described
herein may be confirmed by a number of pharmacological assays as
described below.
[0358] Assay 1: Determination of Glutaminase Enzyme Activity
[0359] Compounds can be assessed for their ability to inhibit the
enzymatic activity of recombinant Glutaminase 1 (GAC) using a
biochemical assay in a 2-step procedure: 1. conversion of
L-glutamine to glutamate by GAC, and 2. glutamate to
alpha-ketoglutarate catalyzed by glutamate dehydrogenase (GDH).
Reduction of NAD to NADH manifested by a change in absorbance is
determined spectrophotometrically. A substrate solution is prepared
(50 mM Tris-HCl pH 8.0, 0.2 mM EDTA, 150 mM K.sub.2HPO.sub.4, 0.1
mg/ml BSA, 1 mM DTT, 20 mM L-glutamine, 2 mM NAD, and 10 ppm
antifoam) and 50 of the substrate solution is to be added to a
96-well half area clear plate. The compound is added as a DMSO
solution. Enzymatic reaction is started by the addition of 50 .mu.l
of enzyme solution (50 mM Tris-HCl pH 8.0, 0.2 mM EDTA, 150 mM
K.sub.2HPO.sub.4, 0.1 mg/ml BSA, 1 mM DTT, 10 ppm antifoam, 4
units/ml GDH, 4 mM adenosine diphosphate, and 4 nM GAC) and read in
a Molecular Devices M5 plate reader at 20.degree. C. The plate
reader is configured to read absorbance (.lamda.=340 nm) in kinetic
mode for 15 minutes. Data is recorded as milli-absorbance units per
minute and slopes are compared to a control compound and a
DMSO-only control on the same plate. Compounds with slopes less
than the DMSO control can be considered as inhibitors and plate
variability is assessed using the control compound. Activity of the
test compound is reported as % inhibition. The data is analyzed
using Graphpad Prism (Graphpad software; San Diego Calif.) for
IC.sub.50 determination.
[0360] Assay 2: Determination of Enzyme Activity Through Estimation
of Ammonia
[0361] L-glutaminase enzyme assay can be performed using a
colorimetric method by quantifying ammonia formation in a
spectrophotometric analysis using Nessler's Reagent. The procedure
is adopted from British Microbiology Research Journal, 4(1),
97-115, 2014, with modification.
[0362] For routine assay 0.1 ml of properly diluted enzyme
(incubated with or without the test compound) is added to 0.4 ml of
0.025 M L-glutamine solution in 0.1 M boric acid borate buffer (pH
8.0). After incubation for 30 minutes at 37.degree. C., the
reaction is stopped by the addition of 0.5 ml of 1N
H.sub.2SO.sub.4. The precipitated protein is removed by
centrifugation and 0.2 ml of supernatant is added to 3.8 ml of
distilled water. Thereafter, 0.5 ml of Nessler's reagent is added,
and the absorbance measured at 400 nm within 1 to 3 minutes. Enzyme
and substrate blanks are included in all assays, and a standard
curve is prepared with ammonium chloride. The enzyme activity is
expressed as unit (U)/ml. One unit of L-glutaminase is defined as
the amount of enzyme that liberates one micromole (.mu.mol) of
ammonia per minute under standard conditions. The specific activity
(sp. activity) is defined as the units of L-glutaminase per
milligram protein. Accordingly, the change in specific activity of
glutaminase in the presence and absence of test compound is
reported.
[0363] Assay 3: Determination of Glutaminase Enzyme Activity Using
Glutaminase Enzyme from Mice Brain/Kidney
Step 1: Preparation of Tissue Homogenates: Male Balb/c mice were
administered 0.28 M ammonium chloride in drinking water for 7 days.
The animals were sacrificed and brain/kidney organs collected on
dry ice. These organs were suspended in a homogenization buffer
containing 20 mM phosphate buffer-pH 7.4, 0.5 mM EDTA, 5 mM
2-mercaptoethanol, 25% glycerol and 0.02% BSA. The tissue was
homogenized and supernatants were stored at -80.degree. C. until
the enzyme assay was performed.
Step-2: Enzyme Assay
[0364] Aim: Compounds were assessed for their ability to inhibit
the enzymatic activity of L-glutaminase present in mice
brain/kidney homogenate.
[0365] Protocol: The assay was performed using a colorimetric
method using Nessler's Reagent by quantifying the amount of ammonia
formed as a by product during the enzymatic conversion of
L-glutamine to glutamate. In the routine assay, 16 .mu.l of tissue
homogenate is added to 33 .mu.l of Tris-Hcl phosphate buffer (pH 8)
along with 1 .mu.l of DMSO/test compound containing the desired
final concentration and vortexed briefly. 50 .mu.l of 20 mM
L-glutamine Tris buffer is added to start the reaction and
incubated for 15 minutes at 37.degree. C. The ammonia formed is
detected by adding 20 .mu.l of reaction mixture to cold water in a
96 well plate followed by 20 .mu.l of Nessler's reagent. The colour
developed is measured at 450 nm. Data Analysis: Activity of the
test compound is reported as % inhibition and the data is analyzed
using Graphpad Prism (Graphpad software; San Diego Calif.) for
IC.sub.50 determination.
[0366] Results for Assay 3:
TABLE-US-00004 Glutaminase(Brain) activity % inhibition Ex. No. @1
.mu.M 0.1 .mu.M IC50 Example 1 B D E Example 2 C D -- Example 2A D
D -- Example 2B D D -- Example 3 B C -- Example 3A D D -- Example
3B D D -- Example-4 B C D Example-5 C D -- Example-6 D D --
Example-7 D D -- Example-8 A A A Example-9 A B B Example-10 A B B
Example-11 B D -- Example-12 A B B Example-13 B D -- Example-14 A A
A Example-15 A B A Example-16 A A A Example-17 A A A Example-18 A A
A Example-19 A B -- Example-20 B C B Example-21 A B B Example-22 A
B B Example-23 A B B Example-24 A B B Example-25 A B B Example-26 A
B B Example-27 A B -- Example-28 B B B Example-29 A B B Example-30
A B B Example-31 B C -- Example-32 B C -- Example-33 B C --
Example-34 B C -- Example-35 A B A Example-36 B C For percent (%)
inhibition: A is >75% to 100%; B is >50% to .ltoreq.75%; C is
>25% to .ltoreq.50%; and D is .ltoreq.25%. For IC50: A is <50
nM; B is .gtoreq.50 nM to <200 nM; C is .gtoreq.200 nM to
<500 nM; D is .gtoreq.500 nM to <1000 nM; and E is
.gtoreq.1000 nM to .ltoreq.2000 nM.
[0367] Assay 4: In Vitro Cell Proliferation Assay in Cancer Cell
Lines
[0368] Growth inhibition assays were carried out using 10% FBS
supplemented media. Cells were seeded at a concentration of
5000-20,000 cells/well in a 96-well plate. Test compounds at a
concentration range from 0.01 to 10000 nM were added after 24
hours. Growth was assessed using the
3-[4,5-dimethylthiazol-2-yl]-2,5-diphenytetrazolium bromide (MTT)
dye reduction test at 0 h (prior to the addition of the test
compound) and 72 hours after the addition of test compound.
Absorbance was read on a Fluostar Optima (BMG Labtech, Germany) at
a wave length of 450 nm. Data were analysed using GraphPad Prism
and percent inhibition due to the test compound compared to the
control was calculated accordingly. The results are as shown
below.
TABLE-US-00005 GI50 Ex. No. MDA-MB-231 A549 HCT116 Daudi Rajii
Example 3 E -- -- -- -- Example-4 C -- -- Example-8 -- B C A A
Example-9 B -- -- -- -- Example-10 B -- -- -- -- Example-11 C -- --
-- -- Example-12 A -- -- -- -- Example-14 A -- -- -- -- Example-15
A -- -- -- -- Example-17 A -- -- -- -- Example-18 A -- -- -- --
GI50: A is <100 nM; B is .gtoreq.100 nM to <250 nM; C is
.gtoreq.250 nM to <500 nM; D is .gtoreq.500 nM to <1000 nM
and E is .gtoreq.1000 nM to .ltoreq.3000 nM.
[0369] Although the invention herein has been described with
reference to particular embodiments, it is to be understood that
these embodiments are merely illustrative of the principles and
applications of the present invention. It is therefore to be
understood that numerous modifications may be made to the
illustrative embodiments and that other arrangements may be devised
without departing from the spirit and scope of the present
invention as described above. It is intended that the appended
claims define the scope of the invention and that methods and
structures within the scope of these claims and their equivalents
be covered thereby.
[0370] All publications and patent and/or patent applications cited
in this application are herein incorporated by reference to the
same extent as if each individual publication or patent application
was specifically and individually indicated to be incorporated
herein by reference.
* * * * *