U.S. patent application number 10/989717 was filed with the patent office on 2005-12-01 for urea derivatives as flt-3 modulators.
This patent application is currently assigned to Ambit Biosciences Corporation. Invention is credited to Grotzfeld, Robert M., Lai, Andiliy G., Lockhart, David J., Mehta, Shamal A., Milanov, Zdravko V., Patel, Hitesh K..
Application Number | 20050267182 10/989717 |
Document ID | / |
Family ID | 34623983 |
Filed Date | 2005-12-01 |
United States Patent
Application |
20050267182 |
Kind Code |
A1 |
Milanov, Zdravko V. ; et
al. |
December 1, 2005 |
Urea derivatives as FLT-3 modulators
Abstract
The invention provides methods and compositions for treating
conditions mediated FLT-3. The invention also provides methods of
using the compounds and/or compositions in the treatment of a
variety of diseases and unwanted conditions in subjects.
Inventors: |
Milanov, Zdravko V.; (San
Diego, CA) ; Mehta, Shamal A.; (San Diego, CA)
; Grotzfeld, Robert M.; (Carlsbad, CA) ; Lai,
Andiliy G.; (San Diego, CA) ; Patel, Hitesh K.;
(Encinitas, CA) ; Lockhart, David J.; (Del Mar,
CA) |
Correspondence
Address: |
WILSON SONSINI GOODRICH & ROSATI
650 PAGE MILL ROAD
PALO ALTO
CA
943041050
|
Assignee: |
Ambit Biosciences
Corporation
|
Family ID: |
34623983 |
Appl. No.: |
10/989717 |
Filed: |
November 15, 2004 |
Related U.S. Patent Documents
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Application
Number |
Filing Date |
Patent Number |
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60520273 |
Nov 13, 2003 |
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|
60527094 |
Dec 3, 2003 |
|
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60531243 |
Dec 18, 2003 |
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60531082 |
Dec 18, 2003 |
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Current U.S.
Class: |
514/378 |
Current CPC
Class: |
C07D 413/12 20130101;
A61P 43/00 20180101; A61P 11/08 20180101; C07D 513/04 20130101;
C07D 417/14 20130101; A61P 35/02 20180101; A61P 11/00 20180101;
A61P 9/10 20180101; C07D 261/14 20130101; Y02A 50/411 20180101;
C07D 261/08 20130101; C07D 417/12 20130101; C07D 277/82 20130101;
C07D 261/18 20130101; C07D 471/04 20130101; Y02A 50/401 20180101;
A61P 35/04 20180101; C07D 495/04 20130101; C07D 487/04 20130101;
A61P 9/00 20180101; A61P 29/00 20180101; A61P 35/00 20180101; Y02A
50/30 20180101; C07D 277/46 20130101; C07D 231/40 20130101; A61K
31/42 20130101; C07D 231/12 20130101; C07D 413/04 20130101; C07D
471/06 20130101; C07D 405/12 20130101; C07D 413/14 20130101 |
Class at
Publication: |
514/378 |
International
Class: |
A61K 031/42 |
Claims
What is claimed is:
1. A method for modulating FLT-3 kinase, said method comprising
administering an effective amount of a FLT-3 modulating compound
corresponding to Formula (IA): 640wherein: M is substituted or
unsubstituted heteroaryl, or substituted or unsubstituted aryl; N
is a substituted or unsubstituted aryl, or substituted or
unsubstituted hetroaryl; and K is 641Y is O or S; each R.sub.k is
independently H, halogen, substituted or unsubstituted alkyl, --OH,
substituted or unsubstituted alkoxy, --OC(O)R.sub.2, --NO.sub.2,
--N(R.sub.2).sub.2, --SR.sub.2, --C(O)R.sub.2, --C(O).sub.2R.sub.2,
--C(O)N(R.sub.2).sub.2, or --N(R.sub.2)C(O)R.sub.2, each R.sub.2 is
independently H, substituted or unsubstituted alkyl, substituted or
unsubstituted cycloalkyl, substituted or unsubstituted
heterocyclyl, substituted or unsubstituted aryl, or substituted or
unsubstituted heteroaryl; or wherein two R.sub.2 groups are linked
together by an optionally substituted alkylene; and each n is
independently 0, 1, 2, 3 or 4; or an active metabolite, or a
pharmaceutically acceptable prodrug, isomer, pharmaceutically
acceptable salt or solvate thereof.
2. The method of claim 1, wherein said compound corresponds to
Formula (IB): 642wherein: each Z is independently C, CR.sub.3, N,
NR.sub.3, O, or S, provided that no more than two Z's are
heteroatoms and wherein no two adjacent Z's are O or S, where
R.sub.3 is H, substituted or unsubstituted alkyl, substituted or
unsubstituted cycloalkyl, substituted or unsubstituted heteroaryl,
or substituted or unsubstituted aryl; and each R.sub.1 is
independently H, halogen, substituted or unsubstituted alkyl,
substituted or unsubstituted alkoxy, substituted or unsubstituted
cycloalkyl, substituted or unsubstituted heterocyclyl, substituted
or unsubstituted aryl, substituted or unsubstituted heteroaryl,
--OR.sub.c --OH, --OC(O)R.sub.c, --NO.sub.2, --N(R.sub.c).sub.2,
--SR.sub.c, S(O).sub.jR.sub.c where j is 1 or 2,
--NR.sub.cC(O)R.sub.c, --C(O)N(R.sub.c).sub.2, C(O).sub.2R.sub.c,
or --C(O)R.sub.c; or two adjacent R.sub.1's, are taken together to
form a substituted or unsubstituted aryl or heteroaryl, where each
R.sub.c is independently H, substituted or unsubstituted alkyl,
substituted or unsubstituted cycloalkyl, substituted or
unsubstituted aryl, or substituted or unsubstituted heteroaryl.
3. The method of claim 2, wherein said compound corresponds to
Formula (I): 643
4. The method of claim 3, wherein said compound corresponds to
Formula (II): 644
5. The method of claim 4, wherein said compound corresponds to
Formula (III): 645wherein Z.sub.1 is CR.sub.3 or N; and Z.sub.2 is
O or S.
6. The method of claim 5, wherein said compound is selected from
the group consisting of: 646
7. The method of claim 5, wherein said compound corresponds to
Formula (IV): 647wherein: each R.sub.1 is independently H, halogen,
substituted or unsubstituted alkyl, --O(substituted or
unsubstituted alkyl), --O(substituted or unsubstituted alkenyl),
--NR.sub.cC(O)O(substituted or unsubstituted alkyl),
--NR.sub.cC(O)(substituted or unsubstituted alkyl),
--NR.sub.cC(O)(substituted or unsubstituted alkenyl),
--C(O)NR.sub.c(substituted or unsubstituted alkyl),
--C(O)NR.sub.c(substituted or unsubstituted alkenyl), --NO.sub.2,
--S(.dbd.O)R.sub.c, --SR.sub.c, C(O).sub.2R.sub.c, or
--C(O)R.sub.c; and each R.sub.2 is independently H or substituted
or unsubstituted alkyl.
8. The method of claim 5, wherein said compound corresponds to
Formula (V): 648
9. The method of claim 8, wherein said compound is selected from
the group consisting of: 649650
10. The method of claim 10, wherein said compound corresponds to
Formula (VII): 651wherein Z.sub.1 is O or S; and Z.sub.2 is
CR.sub.3 or N.
11. The method of claim 1, wherein said compound corresponds to
Formula (VI): 652wherein: each R.sub.1 is independently H, halogen,
substituted or unsubstituted alkyl, --O(substituted or
unsubstituted alkyl), --O(substituted or unsubstituted alkenyl),
--NR.sub.cC(O)O(substituted or unsubstituted alkyl), --NR.sub.cC(O)
(substituted or unsubstituted alkyl), --NR.sub.cC(O)(substituted or
unsubstituted alkenyl), --C(O)NR.sub.c(substituted or unsubstituted
alkyl), --C(O)NR.sub.c(substituted or unsubstituted alkenyl),
--NO.sub.2, --S(.dbd.O)R.sub.c, --SR.sub.c, C(O).sub.2R.sub.c, or
--C(O)R.sub.c; and each R.sub.2 is independently H or substituted
or unsubstituted alkyl.
12. The method of claim 10, wherein said compound corresponds to
Formula (VIII): 653
13. The method of claim 12, wherein said compound is selected from
the group consisting of: 654
14. The method of claim 5, wherein said compound corresponds to
Formula (IX): 655wherein: L is a linker selected from the group
consisting of a covalent bond, substituted or unsubstituted
alkenylene, substituted or unsubstituted alkylene, --C(O)NH--,
--C(O)--, --NH--, --O--, --S--, --O(substituted or unsubstituted
alkylene)-, --N(substituted or unsubstituted alkylene)-,
--C(O)NH(substituted or unsubstituted alkylene)-,
--C(O)NH(substituted or unsubstituted alkenylene)-
--NHC(O)(substituted or unsubstituted alkylene)-,
--NHC(O)(substituted or unsubstituted alkenylene)-,
--C(O)(substituted or unsubstituted alkenylene)-, and
--NHC(O)(substituted or unsubstituted alkylene)S(substituted or
unsubstituted alkylene)C(O)NH--; and T is a mono-, bi, -or
tricyclic, substituted or unsubstituted cycloalkyl, heterocyclyl,
aryl, or heteroaryl.
15. The method of claim 14, wherein said compound is selected from
the group consisting of: 656657
16. The method of claim 14, wherein T of said compound corresponds
to Formula (X): 658wherein A is a substituted or unsubstituted five
or six-membered heterocyclyl, aryl, or heteroaryl; and B is a
substituted or unsubstituted five or six-membered heterocyclene,
arylene, or heteroarylene, wherein A and B together form a fused
two ring moiety.
17. The method of claim 16, wherein said compound corresponds to
Formula (XI): 659
18. The method of claim 17, wherein L of said compound is a
covalent bond, --C(O)NH(substituted or unsubstituted alkylene)-,
--NHC(O)--, --NHC(O)(substituted or unsubstituted alkylene)-,
--NH--, or --O(substituted or unsubstituted alkylene)-.
19. The method of claim 18, wherein said compound corresponds to
Formula (XII): 660
20. The method of claim 19, wherein B of said compound is a
substituted or unsubstituted five-membered arylene or
heteroarylene.
21. The method of claim 20, wherein said compound is: 661
22. The method of claim 20, wherein B is substituted or
unsubstituted thiophenylene.
23. The method of claim 22, wherein said compound is selected from
the group consisting of: 662
24. The method of claim 20, wherein B is substituted or
unsubstituted imidazolylene.
25. The method of claim 24, wherein said compound is selected from
the group consisting of: 663
26. The method of claim 20, wherein B is substituted or
unsubstituted pyrrolylene.
27. The method of claim 26, wherein said compound is selected from
the group consisting of: 664
28. The method of claim 19, wherein B of said compound is a
substituted or unsubstituted 6-membered arylene or
heteroarylene.
29. The method of claim 28, wherein B is substituted or
unsubstituted phenylene.
30. The method of claim 29, wherein said compound is selected from
the group consisting of: 665
31. The method of claim 28, wherein B is substituted or
unsubstituted pyridinylene, pyrimidinylene, or pyridazine.
32. The method of claim 31, wherein said compound is selected from
the group consisting of: 666667
33. The method of claim 18, wherein said compound corresponds to
Formula (XIII): 668
34. The method of claim 33, wherein B of said compound is a
substituted or unsubstituted six-membered heteroarylene.
35. The method of claim 34, wherein said six-membered heteroarylene
is substituted or unsubstituted pyrimidinylene.
36. The method of claim 35, wherein said compound is selected from
the group consisting of: 669
37. The method of claim 18, wherein L of said compound
--OCH.sub.2--.
38. The method of claim 37, wherein said compound is selected from
the group consisting of: 670
39. The method of claim 14, wherein L of said compound is
--C(O)NH.
40. The method of claim 39, wherein said compound is selected from
the group consisting of: 671
41. The method of claim 14, wherein said compound is selected from
the group consisting of: 672
42. The method of claim 5, wherein said compound corresponds to
Formula (XIV): 673wherein: L is a linker selected from the group
consisting of a covalent bond, substituted or unsubstituted
alkenylene, substituted or unsubstituted alkylene, --C(O)NH--,
--C(O)--, --NH--, --O--, --S--, --O(substituted or unsubstituted
alkylene)-, --N(substituted or unsubstituted alkylene)-,
--C(O)NH(substituted or unsubstituted alkylene)-,
--C(O)NH(substituted or unsubstituted alkenylene)-
--NHC(O)(substituted or unsubstituted alkylene)-,
--NHC(O)(substituted or unsubstituted alkenylene)-,
--C(O)(substituted or unsubstituted alkenylene)-, and
--NHC(O)(substituted or unsubstituted alkylene)S(substituted or
unsubstituted alkylene)C(O)NH--; and each of X.sub.1-X.sub.5 is
independently C, CR, N, NR, S, or O, wherein no more than three of
X.sub.1-X.sub.5 is a heteroatom, and no two adjacent ring atoms are
O or S; where each R is independently H, halogen, substituted or
unsubstituted alkyl, --OH, substituted or unsubstituted alkoxy,
--OC(O)R.sub.d, --NO.sub.2, --N(R.sub.d).sub.2, --SR.sub.d,
--S(O).sub.jR.sub.d where j is 1 or 2, --NR.sub.d C(O)R.sub.d,
--C(O).sub.2R.sub.d, --C(O)N(R.sub.d).sub.2 or --C(O)R.sub.d, or
two adjacent R's are taken together to form a substituted or
unsubstituted aryl or hetroaryl, where each R.sub.d is
independently H, substituted or unsubstituted alkyl, substituted or
unsubstituted cycloalkyl, substituted or unsubstituted aryl or
substituted or unsubstituted heteroaryl.
43. The method of claim 42, wherein said compound corresponds to
Formula (XV): 674
44. The method of claim 43, wherein L of said compound is a
covalent bond, --C(O)NH--, or --O(substituted or unsubstituted
alkylene)-.
45. The method of claim 44, wherein 675of said compound is selected
from the group consisting of: 676
46. The method of claim 45, wherein said compound is selected from
the group consisting of: 677678679
47. The method of claim 5, wherein said compound corresponds to
Formula (XVI): 680wherein: L is a linker selected from the group
consisting of a covalent bond, substituted or unsubstituted
alkenylene, substituted or unsubstituted alkylene, --C(O)NH--,
--C(O)--, --NH--, --O--, --S--, --O(substituted or unsubstituted
alkylene)-, --N(substituted or unsubstituted alkylene)-,
--C(O)NH(substituted or unsubstituted alkylene)-,
--C(O)NH(substituted or unsubstituted alkenylene)-
--NHC(O)(substituted or unsubstituted alkylene)-,
--NHC(O)(substituted or unsubstituted alkenylene)-,
--C(O)(substituted or unsubstituted alkenylene)-, and
--NHC(O)(substituted or unsubstituted alkylene)S(substituted or
unsubstituted alkylene)C(O)NH--; and each of X.sub.1-X.sub.5 is
independently C, CR, N--O, or N, wherein no more than two of
X.sub.1-X.sub.5 is N, where each R is independently H, halogen,
substituted or unsubstituted alkyl, --OH, substituted or
unsubstituted alkoxy, --OC(O)R.sub.d, --NO.sub.2,
--N(R.sub.d).sub.2, --SR.sub.d, --S(O).sub.jR.sub.d where j is 1 or
2, --NR.sub.dC(O)R.sub.d, --C(O).sub.2R.sub.d,
--C(O)N(R.sub.d).sub.2 or --C(O)R.sub.d, or two adjacent R's are
taken together to form a substituted or unsubstituted aryl or
hetroaryl, where each R.sub.d is independently H, substituted or
unsubstituted alkyl, substituted or unsubstituted cycloalkyl,
substituted or unsubstituted aryl or substituted or unsubstituted
heteroaryl.
48. The method of claim 47, wherein said compound corresponds to
Formula (XVII): 681
49. The method of claim 48, wherein said compound is: 682
50. The method of claim 48, wherein said compound corresponds to
Formula (XVIII): 683
51. The method of claim 50, wherein said compound is selected from
the group consisting of: 684685
52. The method of claim 48, wherein said compound corresponds to
Formula (XXI): 686
53. The method of claim 52, wherein said compound is selected from
the group consisting of: 687
54. The method of claim 48, wherein said compound corresponds to
Formula (XXII): 688wherein L is --O (substituted or unsubstituted
alkylene)- or --C(O)(substituted or unsubstituted alkenylene)-.
55. The method of claim 54, wherein said compound is selected from
the group consisting of: 689
56. The method of claim 48, wherein said compound corresponds to
Formula (XXIII): 690
57. The method of claim 56, wherein said compound is selected from
the group consisting of: 691
58. The method of claim 56, wherein said compound is selected from
the group consisting of: 692
59. The method of claim 58, wherein said compound is selected from
the group consisting of: 693
60. The method of claim 48, wherein said compound corresponds to
Formula (XXV): 694
61. The method of claim 60, wherein L of said compound is
--O(substituted or unsubstituted alkylene)- or --O(substituted or
unsubstituted alkenylene)-.
62. The method of claim 48, wherein said compound is selected from
the group consisting of: 695696697
63. The method of claim 60, wherein L of said compound is
--NHC(O)--.
64. The method of claim 63, wherein said compound is selected from
the group consisting of: 698699
65. The method of claim 60, wherein L of said compound is a
covalent bond, substituted or unsubstituted alkylene,
--NHC(O)(substituted or unsubstituted alkylene)-,
--NHC(O)(substituted or unsubstituted alkenylene)-,
--NH(alkylene)-, --NHC(O)CH.sub.2SCH.sub.2C(O)NH--, and
--NHC(O)(substituted alkylene)S--.
66. The method of claim 65, wherein said compound is selected from
the group consisting of: 700701
67. The method of claim 10, wherein said compound corresponds to
Formula (XXVI): 702wherein: L is a linker selected from the group
consisting of a covalent bond, substituted or unsubstituted
alkenylene, substituted or unsubstituted alkylene, --C(O)NH--,
--C(O)--, --NH--, --O--, --S--, --O(substituted or unsubstituted
alkylene)-, --N(substituted or unsubstituted alkylene)-,
--C(O)NH(substituted or unsubstituted alkylene)-,
--C(O)NH(substituted or unsubstituted alkenylene)-
--NHC(O)(substituted or unsubstituted alkylene)-,
--NHC(O)(substituted or unsubstituted alkenylene)-,
--C(O)(substituted or unsubstituted alkenylene)-, and
--NHC(O)(substituted or unsubstituted alkylene)S(substituted or
unsubstituted alkylene)C(O)NH--; and each of X.sub.1-X.sub.5 is
independently C, CR, or N, wherein no more than two of X, --X.sub.5
is N, where each R is independently H, halogen, substituted or
unsubstituted alkyl, --OH, substituted or unsubstituted alkoxy,
--OC(O)R.sub.d, --NO.sub.2, --N(R.sub.d).sub.2, --SR.sub.d,
--S(O).sub.jR.sub.d where j is 1 or 2, --NR.sub.d C(O)R.sub.d,
--C(O).sub.2R.sub.d, --C(O)N(R.sub.d).sub.2 or --C(O)R.sub.d, or
two adjacent R's are taken together to form a substituted or
unsubstituted aryl or hetroaryl, where each R.sub.d is
independently H, substituted or unsubstituted alkyl, substituted or
unsubstituted cycloalkyl, substituted or unsubstituted aryl or
substituted or unsubstituted heteroaryl. Z.sub.1 is O or S; and
Z.sub.2 is CR.sub.3 or N.
68. The method of claim 67, wherein said compound corresponds to
Formula (XXVII): 703
69. The method of claim 68, wherein said compound is selected from
the group consisting of; 704
70. The method of claim 5, wherein said compound corresponds to
Formula (XVIII): 705wherein: each of L and L.sub.1 is independently
a linker selected from the group consisting of a covalent bond,
substituted or unsubstituted alkenylene, substituted or
unsubstituted alkylene, --C(O)NH--, --C(O)--, --NH--, --O--, --S--,
--O(substituted or unsubstituted alkylene)-, --N(substituted or
unsubstituted alkylene)-, --C(O)NH(substituted or unsubstituted
alkylene), --C(O)NH(substituted or unsubstituted alkenylene)-
--NHC(O)(substituted or unsubstituted alkylene)-,
--NHC(O)(substituted or unsubstituted alkenylene)-,
--C(O)(substituted or unsubstituted alkenylene)-, and
--NHC(O)(substituted or unsubstituted alkylene)S(substituted or
unsubstituted alkylene)C(O)NH--; U is a substituted or
unsubstituted cycloalkyl, heterocycloalkyl, aryl, or heteroaryl;
and V is a substituted or unsubstituted cycloalkylene,
heterocyclene, arylene, or heteroarylene.
71. The method of claim 70, wherein said compound corresponds to
Formula (XIX): 706
72. The method of claim 71, wherein said compound corresponds to
Formula (XX): 707
73. The method of claim 72, wherein said compound is selected from
the group consisting of: 708709710
74. The method of claim 71, wherein said compound corresponds to
Formula (XXXI): 711wherein: each of X.sub.1-X.sub.5 is
independently C, CR, N, NR, S, or O, wherein no more than three of
X.sub.1-X.sub.5 is a heteroatom, and no two adjacent ring atoms are
O or S; and each R is independently H, halogen, substituted or
unsubstituted alkyl, --OH, substituted or unsubstituted alkoxy,
--OC(O)R.sub.d, --NO.sub.2, --N(R.sub.d).sub.2, --SR.sub.d,
--S(O).sub.jR.sub.d where j is 1 or 2, --NR.sub.d C(O)R.sub.d,
--C(O).sub.2R.sub.d, --C(O)N(R.sub.d).sub.2 or --C(O)R.sub.d, or
two adjacent R's are taken together to form a substituted or
unsubstituted aryl or hetroaryl, where each R.sub.d is
independently H, substituted or unsubstituted alkyl, substituted or
unsubstituted cycloalkyl, substituted or unsubstituted aryl or
substituted or unsubstituted heteroaryl.
76. The method of claim 75, wherein U is a substituted or
unsubstituted five-membered heteroaryl, substituted or
unsubstituted phenyl, or substituted or unsubstituted six-membered
heteroaryl.
77. The method of claim 76, wherein said compound is selected from
the group consisting of: 712713714
78. The method of claim 71, wherein said compound is selected from
the group consisting of: 715716
79. The method of claim 4, wherein said compound corresponds to
Formula (XXIII): 717wherein: Z.sub.3 is NR.sub.8, O, or S; Z.sub.4
is N or CR.sub.8; and R.sub.8 is H, substituted or unsubstituted
alkyl, substituted or unsubstituted alkenyl, substituted or
unsubstituted cycloalkyl, substituted or unsubstituted heteroaryl,
or substituted or unsubstituted aryl.
80. The method of claim 79, wherein said compound corresponds to
Formula (XXIV): 718
81. The method of claim 80, wherein said compound is selected from
the group consisting of: 719
82. The method of claim 80, wherein said compound corresponds to
Formula (XXV) 720wherein: L is a linker selected from the group
consisting of a covalent bond, substituted or unsubstituted
alkenylene, substituted or unsubstituted alkylene, --C(O)NH--,
--C(O)--, --NH--, --O--, --S--, --O(substituted or unsubstituted
alkylene)-, --N(substituted or unsubstituted alkylene)-,
--C(O)NH(substituted or unsubstituted alkylene)-,
--C(O)NH(substituted or unsubstituted alkenylene)-
--NHC(O)(substituted or unsubstituted alkylene)-,
--NHC(O)(substituted or unsubstituted alkenylene)-,
--C(O)(substituted or unsubstituted alkenylene)-, and
--NHC(O)(substituted or unsubstituted alkylene)S(substituted or
unsubstituted alkylene)C(O)NH--; and T is a substituted or
unsubstituted cycloalkyl, heterocycloalkyl, aryl, or
heteroaryl.
83. The method of claim 82, wherein said compound is selected from
the group consisting of: 721
84. The method of claim 1, wherein said compound corresponds to
Formula (A): 722wherein: each Z is independently C, CR.sub.3, N.
NR.sub.3, O, or S, provided that no more than two Z's are
heteroatoms where R.sub.3 is H, substituted or unsubstituted alkyl,
substituted or unsubstituted cycloalkyl, substituted or
unsubstituted heteroaryl, or substituted or unsubstituted aryl.
each R.sub.2 is independently H, substituted or unsubstituted
alkyl, substituted or unsubstituted cycloalkyl, substituted or
unsubstituted heterocyclyl, substituted or unsubstituted aryl, or
substituted or unsubstituted heteroaryl; or wherein two R.sub.2
groups are linked together by an optionally substituted alkylene;
and each R.sub.1 is independently H, halogen, substituted or
unsubstituted alkyl, substituted or unsubstituted alkoxy,
substituted or unsubstituted cycloalkyl, substituted or
unsubstituted heterocyclyl, substituted or unsubstituted aryl,
substituted or unsubstituted heteroaryl, --OR.sub.c --OH,
--OC(O)R.sub.c, --NO.sub.2, --N(R.sub.c).sub.2, --SR.sub.c,
S(O).sub.jR.sub.c where j is 1 or 2, --NR.sub.cC(O)R.sub.c,
--C(O)N(R.sub.c).sub.2, C(O).sub.2R.sub.c, or --C(O)R.sub.c; or two
adjacent R.sub.1 's, are taken together to form a substituted or
unsubstituted aryl or heteroaryl, where each R.sub.c is
independently H, substituted or unsubstituted alkyl, substituted or
unsubstituted cycloalkyl, substituted or unsubstituted aryl, or
substituted or unsubstituted heteroaryl.
85. The method of claim 84, wherein said compound corresponds to
Formula (B): 723
86. The method of claim 85, wherein said compound is selected from
the group consisting of: 724
87. The method of claim 1, wherein said compound is selected from
the group consisting of: 725
88. A method for treating a disease mediated by FLT-3 kinase, said
method comprising administering a therapeutically effective amount
of a compound corresponding to Formula (IA): 726wherein: M is
substituted or unsubstituted heteroaryl, or substituted or
unsubstituted aryl; N is a substituted or unsubstituted aryl, or
substituted or unsubstituted hetroaryl; and K is 727Y is O or S;
each R.sub.k is independently H, halogen, substituted or
unsubstituted alkyl, --OH, substituted or unsubstituted alkoxy,
--OC(O)R.sub.2, --NO.sub.2, --N(R.sub.2).sub.2, --SR.sub.2,
--C(O)R.sub.2, --C(O).sub.2R.sub.2, --C(O)N(R.sub.2).sub.2, or
--N(R.sub.2)C(O)R.sub.2, each R.sub.2 is independently H,
substituted or unsubstituted alkyl, substituted or unsubstituted
cycloalkyl, substituted or unsubstituted heterocyclyl, substituted
or unsubstituted aryl, or substituted or unsubstituted heteroaryl;
or wherein two R.sub.2 groups are linked together by an optionally
substituted alkylene; and each n is independently 0, 1, 2, 3 or 4;
or an active metabolite, or a pharmaceutically acceptable prodrug,
isomer, pharmaceutically acceptable salt or solvate thereof.
Description
[0001] This application claims priority to U.S. Provisional
Application No. 60/520,273, filed Nov. 13, 2003, U.S. Provisional
Application No. 60/527,094, filed Dec. 3, 2003, U.S. Provisional
Application No. 60/531,243, filed Dec. 18, 2003, and U.S.
Provisional Application No. 60/531,082, filed Dec. 18, 2003, the
contents of which are incorporated herein by reference in their
entirety.
BACKGROUND
[0002] Protein-kinases (PKs) play a role in signal transduction
pathways regulating a number of cellular functions, such as cell
growth, differentiation, and cell death. PKs are enzymes that
catalyze the phosphorylation of hydroxy groups on tyrosine, serine
and threonine residues of proteins, and can be conveniently broken
down into two classes, the protein tyrosine kinases (PTKs) and the
serine-threonine kinases (STKs). Growth factor receptors with PTK
activity are known as receptor tyrosine kinases. Protein receptor
tyrosine kinases are a family of tightly regulated enzymes, and the
aberrant activation of various members of the family is one of the
hallmarks of cancer. The protein-tyrosine kinase family, which
includes Bcr-Abl tyrosine kinase, can be divided into subgroups
that have similar structural organization and sequence similarity
within the kinase domain. The members of the type III group of
receptor tyrosine kinases include the platelet-derived growth
factor (PDGF) receptors (PDGF receptors .alpha. and .beta.),
colony-stimulating factor (CSF-1) receptor (CSF-1R, c-Fms), FLT-3,
and stem cell or steel factor receptor (c-kit). A more complete
listing of the known Protein receptor tyrosine kinases subfamilies
is described in Plowman et al., DN&P, 7(6): 334-339 (1994),
which is incorporated by reference, including any drawings, as if
fully set forth herein. Furthermore, for a more detailed discussion
of "non-receptor tyrosine kinases", see Bolen, Oncogene, 8:
2025-2031 (1993), which is incorporated by reference, including any
drawings, as if fully set forth herein.
[0003] Hematologic cancers, also known as hematologic or
hematopoietic malignancies, are cancers of the blood or bone
marrow; including leukemia and lymphoma. Acute myelogenous leukemia
(AML) is a clonal hematopoietic stem cell leukemia that represents
.about.90% of all acute leukemias in adults. See e.g., Lowenberg et
al., N. Eng. J. Med. 341: 1051-62 (1999). While chemotherapy can
result in complete remissions, the long term disease-free survival
rate for AML is about 14% with about 7,400 deaths from AML each
year in the United States. The single most commonly mutated gene in
AML is FLT3 kinase. See e.g., Abu-Duhier et al., Br. J. Haemotol.
111: 190-05 (2000); Kiyoi et al., Blood 93: 3074-80 (1999);
Kottaridis et al., Blood 98: 1752-59 (2001); Stirewalt et al.,
Blood 97: 3589-95 (2001). Such mutations also indicate a poor
prognosis for the patient.
[0004] The compounds provided by the present invention are urea
derivatives of substituted aryls and hetroaryls, e.g., isoxazoles,
pyrazoles and isothiazoles. Urea derivatives of pyrazoles have been
reported to be selective p38 kinase inhibitors by Dumas, J., et
al., Bioorg. Medic. Chem. Lett. 10: 2051-2054 (2000). Oxazoles and
isopyrazoles are suggested as blockers of cytokine production in WO
00/43384 published 27 Jul. 2000. Urea derivatives of isoxazole and
pyrazoles are described as inhibitors of RAF kinase in WO 99/32106
published 1 Jul. 1999. Such compounds are also described as p38
kinase inhibitors by Dumas, J., et al., Bioorg. Medic. Chem. Lett.
10: 2047-2050 (2000). These compounds are also suggested as p38
kinase inhibitors in PCT publication WO 99/32111 published 1 Jul.
1999.
[0005] There remains a need for additional compounds that are
effective in inhibiting kinase activity. Given the complexities of
signal transduction with the redundancy and crosstalk between
various pathways, the identification of specific kinase inhibitors
permits accurate targeting with limited inhibition of other
pathways, thus reducing the toxicity of such inhibitory
compounds.
SUMMARY OF THE INVENTION
[0006] The present invention provides compounds which modulate
kinase activity, and in some embodiments inhibit protein tyrosine
kinases or a specific kinase or kinase class. In some embodiments,
the compositions and methods for treating and preventing conditions
and diseases, such as cancer, hematologic malignancies,
cardiovascular disease, inflammation or multiple sclerosis. The
compounds of the invention can be delivered alone or in combination
with additional agents, and are used for the treatment and/or
prevention of conditions and diseases. Unless otherwise stated,
each of the substituents is as previously defined.
[0007] Provided herein are compositions and methods for treating a
disease comprising administering to a subject in need thereof an
effective amount of a FLT-3 modulating compound having the
structure: 1
[0008] wherein:
[0009] (a) R.sub.3a and R.sub.4a are each a suitable substituent
independently selected from hydrogen, or an alkyl, alkenyl,
heteroalkyl, haloalkyl, alkynyl, aryl, cycloalkyl,
heterocycloalkyl, heteroaryl group unsubstituted or substituted
with one or more suitable substituents independently selected from
the group consisting of: halogens; --CN; and --NO.sub.2; and alkyl,
alkenyl, heteroalkyl, haloalkyl, alkynyl, aryl, cycloalkyl,
heterocycloalkyl, heteroaryl, --(CH.sub.2).sub.zCN where z is a
whole integer, preferably from 0 to 4, .dbd.NH, --NHOH, --OH,
--C(O)H, --OC(O)H, --C(O)OH, --OC(O)OH, --OC(O)OC(O)H, --OOH,
--C(NH)NH.sub.2, --NHC(NH)NH.sub.2, --C(S)NH.sub.2,
--NHC(S)NH.sub.2, --NHC(O)NH.sub.2, --S(O.sub.2)H, --S(O)H,
--NH.sub.2, --C(O)NH.sub.2, --OC(O)NH.sub.2, --NHC(O)H, --NHC(O)OH,
--C(O)NHC(O)H, --OS(O.sub.2)H, --OS(O)H, --OSH, --SC(O)H,
--S(O)C(O)OH, --SO.sub.2C(O)OH, --NHSH, --NHS(O)H, --NHSO.sub.2H,
--C(O)SH, --C(O)S(O)H, --C(O)S(O.sub.2)H, --C(S)H, --C(S)OH,
--C(SO)OH, --C(SO.sub.2)OH, --NHC(S)H, --OC(S)H, --OC(S)OH,
--OC(SO.sub.2)H, --S(O.sub.2)NH.sub.2, --S(O)NH.sub.2, --SNH.sub.2,
--NHCS(O.sub.2)H, --NHC(SO)H, --NHC(S)H, and --SH groups
unsubstituted or substituted with one or more suitable substituents
independently selected from the group consisting of halogens,
.dbd.O, --NO.sub.2, --CN, --(CH.sub.2).sub.z--CN where z is a whole
integer, preferably from 0 to 4, --OR.sub.c, --NR.sub.cOR.sub.c,
--NR.sub.cR.sub.c, --C(O)NR.sub.c, --C(O)OR.sub.c, --C(O)R.sub.c,
--NR.sub.cC(O)NR.sub.cR.sub.c, --NR.sub.cC(O)R.sub.c,
--OC(O)OR.sub.c, --OC(O)NR.sub.cR.sub.c, --SR.sub.c, unsubstituted
alkyl, unsubstituted alkenyl, unsubstituted alkynyl, unsubstituted
aryl, unsubstituted cycloalkyl, unsubstituted heterocycloalkyl, and
unsubstituted heteroaryl, or two or more substituents cyclize to
form a fused or spiro polycyclic cycloalkyl, heterocycloalkyl,
aryl, or heteroaryl group, where each R.sub.c is indepenently
selected from hydrogen, unsubstituted alkyl, unsubstituted alkenyl,
unsubstituted alkynyl, unsubstituted aryl, unsubstituted
cycloalkyl, unsubstituted heterocycloalkyl, and unsubstituted
heteroaryl, or two or more R.sub.c groups together cyclize to form
part of a heteroaryl or heterocycloalkyl group unsubstituted or
substituted with an unsubstituted alkyl group;
[0010] or where R.sub.3a and R.sub.4a together cyclize to form part
of a heteroaryl or heterocycloalkyl group unsubstituted or
substituted with one or more suitable substituents selected from
halogen, .dbd.O; .dbd.S; --CN; --NO.sub.2, or an alkyl, alkenyl,
heteroalkyl, haloalkyl, alkynyl, aryl, cycloalkyl,
heterocycloalkyl, heteroaryl group unsubstituted or substituted
with one or more suitable substituents independently selected from
the group consisting of: halogens; .dbd.O; .dbd.S; --CN; and
--NO.sub.2; and alkyl, alkenyl, heteroalkyl, haloalkyl, alkynyl,
aryl, cycloalkyl, heterocycloalkyl, heteroaryl,
--(CH.sub.2).sub.zCN where z is a whole integer, preferably from 0
to 4, .dbd.NH, --NHOH, --OH, --C(O)H, --OC(O)H, --C(O)OH,
--OC(O)OH, --OC(O)OC(O)H, --OOH, --C(NH)NH.sub.2,
--NHC(NH)NH.sub.2, --C(S)NH.sub.2, --NHC(S)NH.sub.2,
--NHC(O)NH.sub.2, --S(O.sub.2)H, --S(O)H, --NH.sub.2,
--C(O)NH.sub.2, --OC(O)NH.sub.2, --NHC(O)H, --NHC(O)OH,
--C(O)NHC(O)H, --OS(O.sub.2)H, --OS(O)H, --OSH, --SC(O)H,
--S(O)C(O)OH, --SO.sub.2C(O)OH, --NHSH, --NHS(O)H, --NHSO.sub.2H,
--C(O)SH, --C(O)S(O)H, --C(O)S(O.sub.2)H, --C(S)H, --C(S)OH,
--C(SO)OH, --C(SO.sub.2)OH, --NHC(S)H, --OC(S)H, --OC(S)OH,
--OC(SO.sub.2)H, --S(O.sub.2)NH.sub.2, --S(O)NH.sub.2, --SNH.sub.2,
--NHCS(O.sub.2)H, --NHC(SO)H, --NHC(S)H, and --SH groups
unsubstituted or substituted with one or more suitable substituents
independently selected from the group consisting of halogens,
.dbd.O, --NO.sub.2, --CN, --(CH.sub.2).sub.z--CN where z is a whole
integer, preferably from 0 to 4, --OR.sub.c, --NR.sub.cOR.sub.c,
--NR.sub.cR.sub.c, --C(O)NR.sub.c, --C(O)OR.sub.c, --C(O)R.sub.c,
--NR.sub.cC(O)NR.sub.cR.sub.c, --NR.sub.cC(O)R.sub.c,
--OC(O)OR.sub.c, --OC(O)NR.sub.cR.sub.c, --SR.sub.c, unsubstituted
alkyl, unsubstituted alkenyl, unsubstituted alkynyl, unsubstituted
aryl, unsubstituted cycloalkyl, unsubstituted heterocycloalkyl, and
unsubstituted heteroaryl, or two or more substituents cyclize to
form a fused or spiro polycyclic cycloalkyl, heterocycloalkyl,
aryl, or heteroaryl group, where each R.sub.c is independently
selected from hydrogen, unsubstituted alkyl, unsubstituted alkenyl,
unsubstituted alkynyl, unsubstituted aryl, unsubstituted
cycloalkyl, unsubstituted heterocycloalkyl, and unsubstituted
heteroaryl, or two or more R.sub.c groups together cyclize to form
part of a heteroaryl or heterocycloalkyl group unsubstituted or
substituted with an unsubstituted alkyl group;
[0011] (b) Ar.sub.1, Ar.sub.2 and Ar.sub.3 are each independently
an aryl, heteroaryl, cycloalkyl or heterocycloalkyl group
unsubstituted or substituted with one or more suitable substituents
independently selected from the group consisting of: halogens;
.dbd.O; .dbd.S; --CN; and --NO.sub.2; and alkyl, alkenyl,
heteroalkyl, haloalkyl, alkynyl, aryl, cycloalkyl,
heterocycloalkyl, heteroaryl, --(CH.sub.2).sub.zCN where z is a
whole integer, preferably from 0 to 4, .dbd.NH, --NHOH, --OH,
--C(O)H, --OC(O)H, --C(O)OH, --OC(O)OH, --OC(O)OC(O)H, OOH,
--C(NH)NH.sub.2, --NHC(NH)NH.sub.2, --C(S)NH.sub.2,
--NHC(S)NH.sub.2, --NHC(O)NH.sub.2, --S(O.sub.2)H, --S(O)H,
--NH.sub.2, --C(O)NH.sub.2, --OC(O)NH.sub.2, --NHC(O)H, --NHC(O)OH,
--C(O)NHC(O)H, --OS(O.sub.2)H, --OS(O)H, --OSH, --SC(O)H,
--S(O)C(O)OH, --SO.sub.2C(O)OH, --NHSH, --NHS(O)H, --NHSO.sub.2H,
--C(O)SH, --C(O)S(O)H, --C(O)S(O.sub.2)H, --C(S)H, --C(S)OH,
--C(SO)OH, --C(SO.sub.2)OH, --NHC(S)H, --OC(S)H, --OC(S)OH,
--OC(SO.sub.2)H, --S(O.sub.2)NH.sub.2, --S(O)NH.sub.2, --SNH.sub.2,
--NHCS(O.sub.2)H, --NHC(SO)H, --NHC(S)H, and --SH groups
unsubstituted or substituted with one or more suitable substituents
independently selected from the group consisting of halogens,
.dbd.O, --NO.sub.2, --CN, --(CH.sub.2), --CN where z is a whole
integer, preferably from 0 to 4, --OR.sub.c, --NR.sub.cOR.sub.c,
--NR.sub.cR.sub.c, --C(O)NR.sub.c, --C(O)OR.sub.c, --C(O)R.sub.c,
--NR.sub.cC(O)NR.sub.cR.sub.c, --NR.sub.cC(O)R.sub.c,
--OC(O)OR.sub.c, --OC(O)NR.sub.cR.sub.c, --SR.sub.c, unsubstituted
alkyl, unsubstituted alkenyl, unsubstituted alkynyl, unsubstituted
aryl, unsubstituted cycloalkyl, unsubstituted heterocycloalkyl, and
unsubstituted heteroaryl, or two or more substituents cyclize to
form a fused or spiro polycyclic cycloalkyl, heterocycloalkyl,
aryl, or heteroaryl group, where each R.sub.c is independently
selected from hydrogen, unsubstituted alkyl, unsubstituted alkenyl,
unsubstituted alkynyl, unsubstituted aryl, unsubstituted
cycloalkyl, unsubstituted heterocycloalkyl, and unsubstituted
heteroaryl, or two or more R.sub.c groups together cyclize to form
part of a heteroaryl or heterocycloalkyl group unsubstituted or
substituted with an unsubstituted alkyl group;
[0012] (c) n.sub.1 is 0, 1, 2, 3 or 4;
[0013] (d) n.sub.2 is 0, 1, 2, 3 or 4;
[0014] (e) n.sub.3 is 0, 1, 2, 3 or 4;
[0015] (f) Z.sub.a is a bond or is selected from S, O, N, NR.sub.c,
C(O)NR.sub.c, NR.sub.cC(O), and CR.sub.c, wherein R.sub.c is a
suitable substituent selected from hydrogen, unsubstituted alkyl,
unsubstituted alkenyl, unsubstituted alkynyl, unsubstituted aryl,
unsubstituted cycloalkyl, unsubstituted heterocycloalkyl, or
unsubstituted heteroaryl group; and
[0016] (g) W.sub.a is S or O;
[0017] or a pharmaceutically acceptable salt, pharmaceutically
acceptable N-oxide, pharmaceutically active metabolite,
pharmaceutically acceptable prodrug, isomer derivative, or
pharmaceutically acceptable solvate thereof.
[0018] Provided herein are compositions and methods for treating a
disease comprising administering to a subject in need thereof an
effective amount of a FLT-3 modulating compound having the
structure: 2
[0019] wherein:
[0020] M is substituted or unsubstituted heteroaryl, or substituted
or unsubstituted aryl;
[0021] N is a substituted or unsubstituted aryl, or substituted or
unsubstituted hetroaryl; and
[0022] K is 3
[0023] Y is O or S;
[0024] each R.sub.k is independently H, halogen, substituted or
unsubstituted alkyl, --OH, substituted or unsubstituted alkoxy,
--OC(O)R.sub.2, --NO.sub.2, --N(R.sub.2).sub.2, --SR.sub.2,
--C(O)R.sub.2, --C(O).sub.2R.sub.2, --C(O)N(R.sub.2).sub.2, or
--N(R.sub.2)C(O)R.sub.2,
[0025] each R.sub.2 is independently H, substituted or
unsubstituted alkyl, substituted or unsubstituted cycloalkyl,
substituted or unsubstituted heterocyclyl, substituted or
unsubstituted aryl, or substituted or unsubstituted heteroaryl; or
wherein two R.sub.2 groups are linked together by an optionally
substituted alkylene; and
[0026] each n is independently 0, 1, 2, 3 or 4;
[0027] or an active metabolite, or a pharmaceutically acceptable
prodrug, isomer, pharmaceutically acceptable salt or solvate
thereof.
[0028] Provided herein are compositions and methods for treating a
disease comprising administering to a subject in need thereof an
effective amount of a FLT-3 modulating compound having the
structure: 4
[0029] wherein:
[0030] each Z is independently C, CR.sub.3, N, NR.sub.3, O, or S,
provided that no more than two Z's are heteroatoms and wherein no
two adjacent Z's are O or S,
[0031] where R.sub.3 is H, substituted or unsubstituted alkyl,
substituted or unsubstituted cycloalkyl, substituted or
unsubstituted heteroaryl, or substituted or unsubstituted aryl;
and
[0032] each R.sub.1 is independently H, halogen, substituted or
unsubstituted alkyl, substituted or unsubstituted alkoxy,
substituted or unsubstituted cycloalkyl, substituted or
unsubstituted heterocyclyl, substituted or unsubstituted aryl,
substituted or unsubstituted heteroaryl, --OR.sub.c --OH,
--OC(O)R.sub.c, --NO.sub.2, --N(R.sub.c).sub.2, --SR.sub.c,
S(O).sub.jR.sub.c where j is 1 or 2, --NR.sub.cC(O)R.sub.c,
--C(O)N(R.sub.c).sub.2, C(O).sub.2R.sub.c, or --C(O)R.sub.c; or two
adjacent R.sub.1's, are taken together to form a substituted or
unsubstituted aryl or heteroaryl, where
[0033] each R.sub.c is independently H, substituted or
unsubstituted alkyl, substituted or unsubstituted cycloalkyl,
substituted or unsubstituted aryl, or substituted or unsubstituted
heteroaryl.
[0034] Provided herein are compositions and methods for treating a
disease comprising administering to a subject in need thereof an
effective amount of a FLT-3 modulating compound having the
structure: 5
[0035] Provided herein are compositions and methods for treating a
disease comprising administering to a subject in need thereof an
effective amount of a FLT-3 modulating compound having the
structure: 6
[0036] Provided herein are compositions and methods for treating a
disease comprising administering to a subject in need thereof an
effective amount of a FLT-3 modulating compound having the
structure: 7
[0037] wherein Z.sub.1 is CR.sub.3 or N; and Z.sub.2 is O or S.
[0038] Provided herein are compositions and methods for treating a
disease comprising administering to a subject in need thereof an
effective amount of a FLT-3 modulating compound having the
structure: 8
[0039] wherein:
[0040] each R.sub.1 is independently H, halogen, substituted or
unsubstituted alkyl, --O(substituted or unsubstituted alkyl),
--O(substituted or unsubstituted alkenyl),
--NR.sub.c(O)O(substituted or unsubstituted alkyl), --NR.sub.cC(O)
(substituted or unsubstituted alkyl), --NR.sub.cC(O)(substituted or
unsubstituted alkenyl), --C(O)NR.sub.c(substituted or unsubstituted
alkyl), --C(O)NR.sub.c(substituted or unsubstituted alkenyl),
--NO.sub.2, --S(.dbd.O)R.sub.c, --SR.sub.c, C(O).sub.2R.sub.c, or
--C(O)R.sub.c; and
[0041] each R.sub.2 is independently H or substituted or
unsubstituted alkyl.
[0042] Provided herein are compositions and methods for treating a
disease comprising administering to a subject in need thereof an
effective amount of a FLT-3 modulating compound having the
structure: 9
[0043] Provided herein are compositions and methods for treating a
disease comprising administering to a subject in need thereof an
effective amount of a FLT-3 modulating compound having the
structure: 10
[0044] wherein Z, is O or S; and Z.sub.2 is CR.sub.3 or N.
[0045] Provided herein are compositions and methods for treating a
disease comprising administering to a subject in need thereof an
effective amount of a FLT-3 modulating compound having the
structure: 11
[0046] wherein:
[0047] each R.sub.1 is independently H, halogen, substituted or
unsubstituted alkyl, --O(substituted or unsubstituted alkyl),
--O(substituted or unsubstituted alkenyl),
--NR.sub.cC(O)O(substituted or unsubstituted alkyl),
--NR.sub.cC(O)(substituted or unsubstituted alkyl),
--NR.sub.cC(O)(substituted or unsubstituted alkenyl),
--C(O)NR.sub.c(substituted or unsubstituted alkyl),
--C(O)NR.sub.c(substituted or unsubstituted alkenyl), --NO.sub.2,
--S(.dbd.O)R.sub.c, --SR.sub.c, C(O).sub.2R.sub.c, or
--C(O)R.sub.c; and
[0048] each R.sub.2 is independently H or substituted or
unsubstituted alkyl.
[0049] Provided herein are compositions and methods for treating a
disease comprising administering to a subject in need thereof an
effective amount of a FLT-3 modulating compound having the
structure: 12
[0050] Provided herein are compositions and methods for treating a
disease comprising administering to a subject in need thereof an
effective amount of a FLT-3 modulating compound having the
structure: 13
[0051] wherein L is a linker selected from the group consisting of
a covalent bond, substituted or unsubstituted alkenylene,
substituted or unsubstituted alkylene, --C(O)NH--, --C(O)--,
--NH--, --O--, --S--, --O(substituted or unsubstituted alkylene)-,
--N(substituted or unsubstituted alkylene)-, --C(O)NH(substituted
or unsubstituted alkylene)-, --C(O)NH(substituted or unsubstituted
alkenylene)-, --NHC(O)(substituted or unsubstituted alkylene)-,
--NHC(O)(substituted or unsubstituted alkenylene)-,
--C(O)(substituted or unsubstituted alkenylene)-, and
--NHC(O)(substituted or unsubstituted alkylene)S(substituted or
unsubstituted alkylene)C(O)NH--; and T is a mono-, bi-, or
tricyclic, substituted or unsubstituted cycloalkyl, heterocyclyl,
aryl, or heteroaryl. In some embodiments, T is 14
[0052] wherein A is a substituted or unsubstituted five or
six-membered heterocyclyl, aryl, or heteroaryl; and B is a
substituted or unsubstituted five or six-membered heterocyclene,
arylene, or heteroarylene, wherein A and B together form a fused
two ring moiety.
[0053] Provided herein are compositions and methods for treating a
disease comprising administering to a subject in need thereof an
effective amount of a FLT-3 modulating compound having the
structure: 15
[0054] In some embodiments, L of said compound is a covalent bond
--C(O)NH(substituted or unsubstituted alkylene)-, --NHC(O)--,
--NHC(O)(substituted or unsubstituted alkylene)-, --NH--, or
--O(substituted or unsubstituted alkylene)-.
[0055] Provided herein are compositions and methods for treating a
disease comprising administering to a subject in need thereof an
effective amount of a FLT-3 modulating compound having the
structure: 16
[0056] In some embodiments, B of said compound is a substituted or
unsubstituted five-membered arylene or heteroarylene. In other
embodiments, B is substituted or unsubstituted thiophenylene. In
still other embodiments, B is substituted or unsubstituted
imidazolylene. In yet other embodiments, B is substituted or
unsubstituted pyrrolylene. In still other embodiments, B of said
compound is a substituted or unsubstituted 6-membered arylene or
heteroarylene. In some embodiments, B is substituted or
unsubstituted phenylene. In other embodiments, B is substituted or
unsubstituted pyridinylene, pyrimidinylene, or pyridazine.
[0057] Provided herein are compositions and methods for treating a
disease comprising administering to a subject in need thereof an
effective amount of a FLT-3 modulating compound having the
structure: 17
[0058] In some embodiments, B of said compound is a substituted or
unsubstituted six-membered heteroarylene. In other embodiments,
said six-membered heteroarylene is substituted or unsubstituted
pyrimidinylene. In still other embodiments, L of said compound
--OCH.sub.2--. In yet other embodiments, L of said compound is
--C(O)NH.
[0059] Provided herein are compositions and methods for treating a
disease comprising administering to a subject in need thereof an
effective amount of a FLT-3 modulating compound having the
structure: 18
[0060] wherein:
[0061] L is a linker selected from the group consisting of a
covalent bond, substituted or unsubstituted alkenylene, substituted
or unsubstituted alkylene, --C(O)NH--, --C(O)--, --NH--, --O--,
--S--, --O(substituted or unsubstituted alkylene)-, --N(substituted
or unsubstituted alkylene)-, --C(O)NH(substituted or unsubstituted
alkylene)-, --C(O)NH(substituted or unsubstituted alkenylene)-
--NHC(O)(substituted or unsubstituted alkylene)-,
--NHC(O)(substituted or unsubstituted alkenylene)-,
--C(O)(substituted or unsubstituted alkenylene)-, and
--NHC(O)(substituted or unsubstituted alkylene)S(substituted or
unsubstituted alkylene)C(O)NH--; and
[0062] each of X.sub.1-X.sub.5 is independently C, CR, N, NR, S, or
O, wherein no more than three of X.sub.1-X.sub.5 is a heteroatom,
and no two adjacent ring atoms are O or S; where each R is
independently H, halogen, substituted or unsubstituted alkyl, --OH,
substituted or unsubstituted alkoxy, --OC(O)R.sub.d, --NO.sub.2,
--N(R.sub.d).sub.2, --SR.sub.d, --S(O).sub.jR.sub.d where j is 1 or
2, --NR.sub.d C(O)R.sub.d, --C(O).sub.2R.sub.d,
--C(O)N(R.sub.d).sub.2 or --C(O)R.sub.d, or two adjacent R's are
taken together to form a substituted or unsubstituted aryl or
hetroaryl, where
[0063] each R.sub.d is independently H, substituted or
unsubstituted alkyl, substituted or unsubstituted cycloalkyl,
substituted or unsubstituted aryl or substituted or unsubstituted
heteroaryl.
[0064] Provided herein are compositions and methods for treating a
disease comprising administering to a subject in need thereof an
effective amount of a FLT-3 modulating compound having the
structure: 19
[0065] In some embodiments, L of said compound is a covalent bond,
--C(O)NH--, or --O(substituted or unsubstituted alkylene)-. In
other embodiments, 20
[0066] of said compound is selected from the group consisting of:
21
[0067] Provided herein are compositions and methods for treating a
disease comprising administering to a subject in need thereof an
effective amount of a FLT-3 modulating compound having the
structure: 22
[0068] wherein:
[0069] L is a linker selected from the group consisting of a
covalent bond, substituted or unsubstituted alkenylene, substituted
or unsubstituted alkylene, --C(O)NH--, --C(O)--, --NH--, --O--,
--S--, --O(substituted or unsubstituted alkylene)-, --N(substituted
or unsubstituted alkylene)-, --C(O)NH(substituted or unsubstituted
alkylene)-, --C(O)NH(substituted or unsubstituted alkenylene)-
--NHC(O)(substituted or unsubstituted alkylene)-,
--NHC(O)(substituted or unsubstituted alkenylene)-,
--C(O)(substituted or unsubstituted alkenylene)-, and
--NHC(O)(substituted or unsubstituted alkylene)S(substituted or
unsubstituted alkylene)C(O)NH--; and
[0070] each of X.sub.1-X.sub.5 is independently C, CR, N--O, or N,
wherein no more than two of X.sub.1-X.sub.5 is N, where
[0071] each R is independently H, halogen, substituted or
unsubstituted alkyl, --OH, substituted or unsubstituted alkoxy,
--OC(O)R.sub.d, --NO.sub.2, --N(R.sub.d).sub.2, --SR.sub.d,
--S(O).sub.jR.sub.d where j is 1 or 2, --NR.sub.d C(O)R.sub.d,
--C(O).sub.2R.sub.d, --C(O)N(R.sub.d).sub.2 or --C(O)R.sub.d, or
two adjacent R's are taken together to form a substituted or
unsubstituted aryl or hetroaryl, where
[0072] each R.sub.d is independently H, substituted or
unsubstituted alkyl, substituted or unsubstituted cycloalkyl,
substituted or unsubstituted aryl or substituted or unsubstituted
heteroaryl.
[0073] Provided herein are compositions and methods for treating a
disease comprising administering to a subject in need thereof an
effective amount of a FLT-3 modulating compound having the
structure: 23
[0074] Provided herein are compositions and methods for treating a
disease comprising administering to a subject in need thereof an
effective amount of a FLT-3 modulating compound having the
structure: 24
[0075] Provided herein are compositions and methods for treating a
disease comprising administering to a subject in need thereof an
effective amount of a FLT-3 modulating compound having the
structure: 25
[0076] Provided herein are compositions and methods for treating a
disease comprising administering to a subject in need thereof an
effective amount of a FLT-3 modulating compound having the
structure: 26
[0077] wherein L is --O(substituted or unsubstituted alkylene)- or
--C(O)(substituted or unsubstituted alkenylene)-.
[0078] Provided herein are compositions and methods for treating a
disease comprising administering to a subject in need thereof an
effective amount of a FLT-3 modulating compound having the
structure: 27
[0079] Provided herein are compositions and methods for treating a
disease comprising administering to a subject in need thereof an
effective amount of a FLT-3 modulating compound having the
structure: 28
[0080] Provided herein are compositions and methods for treating a
disease comprising administering to a subject in need thereof an
effective amount of a FLT-3 modulating compound having the
structure: 29
[0081] In some embodiments, L of said compound is --O(substituted
or unsubstituted alkylene)- or --O(substituted or unsubstituted
alkenylene)-. In other embodiments, L of said compound is
--NHC(O)--. In still other embodiments, L of said compound is a
covalent bond, substituted or unsubstituted alkylene,
--NHC(O)(substituted or unsubstituted alkylene)-,
--NHC(O)(substituted or unsubstituted alkenylene)-,
--NH(alkylene)-, --NHC(O)CH.sub.2SCH.sub.2C(O)NH--, and
--NHC(O)(substituted alkylene)S--.
[0082] Provided herein are compositions and methods for treating a
disease comprising administering to a subject in need thereof an
effective amount of a FLT-3 modulating compound having the
structure: 30
[0083] wherein:
[0084] L is a linker selected from the group consisting of a
covalent bond, substituted or unsubstituted alkenylene, substituted
or unsubstituted alkylene, --C(O)NH--, --C(O)--, --NH--, --O--,
--S--, --O(substituted or unsubstituted alkylene)-, --N(substituted
or unsubstituted alkylene)-, --C(O)NH(substituted or unsubstituted
alkylene)-, --C(O)NH(substituted or unsubstituted alkenylene)-
--NHC(O)(substituted or unsubstituted alkylene)-,
--NHC(O)(substituted or unsubstituted alkenylene)-,
--C(O)(substituted or unsubstituted alkenylene)-, and
--NHC(O)(substituted or unsubstituted alkylene)S(substituted or
unsubstituted alkylene)C(O)NH--; and
[0085] each of X.sub.1-X.sub.5 is independently C, CR, or N,
wherein no more than two of X.sub.1-X.sub.5 is N, where
[0086] each R is independently H, halogen, substituted or
unsubstituted alkyl, --OH, substituted or unsubstituted alkoxy,
--OC(O)R.sub.d, --NO.sub.2, --N(R.sub.d).sub.2, --SR.sub.d,
--S(O).sub.jR.sub.d where j is 1 or 2, --NR.sub.d C(O)R.sub.d,
--C(O).sub.2R.sub.d, --C(O)N(R.sub.d).sub.2 or --C(O)R.sub.d, or
two adjacent R's are taken together to form a substituted or
unsubstituted aryl or hetroaryl, where
[0087] each R.sub.d is independently H, substituted or
unsubstituted alkyl, substituted or unsubstituted cycloalkyl,
substituted or unsubstituted aryl or substituted or unsubstituted
heteroaryl.
[0088] Z.sub.1 is O or S; and
[0089] Z.sub.2 is CR.sub.3 or N.
[0090] Provided herein are compositions and methods for treating a
disease comprising administering to a subject in need thereof an
effective amount of a FLT-3 modulating compound having the
structure: 31
[0091] Provided herein are compositions and methods for treating a
disease comprising administering to a subject in need thereof an
effective amount of a FLT-3 modulating compound having the
structure: 32
[0092] wherein:
[0093] each of L and L.sub.1 is independently a linker selected
from the group consisting of a covalent bond, substituted or
unsubstituted alkenylene, substituted or unsubstituted alkylene,
--C(O)NH--, --C(O)--, --NH--, --O--, --S--, --O(substituted or
unsubstituted alkylene)-, --N(substituted or unsubstituted
alkylene)-, --C(O)NH(substituted or unsubstituted alkylene),
--C(O)NH(substituted or unsubstituted alkenylene)-
--NHC(O)(substituted or unsubstituted alkylene)-,
--NHC(O)(substituted or unsubstituted alkenylene)-,
--C(O)(substituted or unsubstituted alkenylene)-, and
--NHC(O)(substituted or unsubstituted alkylene)S(substituted or
unsubstituted alkylene)C(O)NH--;
[0094] U is a substituted or unsubstituted cycloalkyl,
heterocycloalkyl, aryl, or heteroaryl; and
[0095] V is a substituted or unsubstituted cycloalkylene,
heterocyclene, arylene, or heteroarylene.
[0096] Provided herein are compositions and methods for treating a
disease comprising administering to a subject in need thereof an
effective amount of a FLT-3 modulating compound having the
structure: 33
[0097] Provided herein are compositions and methods for treating a
disease comprising administering to a subject in need thereof an
effective amount of a FLT-3 modulating compound having the
structure: 34
[0098] Provided herein are compositions and methods for treating a
disease comprising administering to a subject in need thereof an
effective amount of a FLT-3 modulating compound having the
structure: 35
[0099] wherein:
[0100] each of X.sub.1-X.sub.5 is independently C, CR, N, NR, S, or
O, wherein no more than three of X.sub.1-X.sub.5 is a heteroatom,
and no two adjacent ring atoms are O or S; and
[0101] each R is independently H, halogen, substituted or
unsubstituted alkyl, --OH, substituted or unsubstituted alkoxy,
--OC(O)R.sub.d, --NO.sub.2, --N(R.sub.d).sub.2, --SR.sub.d,
--S(O).sub.jR.sub.d where j is 1 or 2, --NR.sub.d C(O)R.sub.d,
--C(O).sub.2R.sub.d, --C(O)N(R.sub.d).sub.2 or --C(O)R.sub.d, or
two adjacent R's are taken together to form a substituted or
unsubstituted aryl or hetroaryl, where
[0102] each R.sub.d is independently H, substituted or
unsubstituted alkyl, substituted or unsubstituted cycloalkyl,
substituted or unsubstituted aryl or substituted or unsubstituted
heteroaryl.
[0103] In some embodiments, U is a substituted or unsubstituted
five-membered heteroaryl, substituted or unsubstituted phenyl, or
substituted or unsubstituted six-membered heteroaryl.
[0104] Provided herein are compositions and methods for treating a
disease comprising administering to a subject in need thereof an
effective amount of a FLT-3 modulating compound having the
structure: 36
[0105] wherein:
[0106] Z.sub.3 is NR.sub.8, O, or S;
[0107] Z.sub.4 is N or CR.sub.8; and
[0108] R.sub.8 is H, substituted or unsubstituted alkyl,
substituted or unsubstituted alkenyl, substituted or unsubstituted
cycloalkyl, substituted or unsubstituted heteroaryl, or substituted
or unsubstituted aryl.
[0109] Provided herein are compositions and methods for treating a
disease comprising administering to a subject in need thereof an
effective amount of a FLT-3 modulating compound having the
structure: 37
[0110] Provided herein are compositions and methods for treating a
disease comprising administering to a subject in need thereof an
effective amount of a FLT-3 modulating compound having the
structure: 38
[0111] wherein:
[0112] L is a linker selected from the group consisting of a
covalent bond, substituted or unsubstituted alkenylene, substituted
or unsubstituted alkylene, --C(O)NH--, --C(O)--, --NH--, --O--,
--S--, --O(substituted or unsubstituted alkylene)-, --N(substituted
or unsubstituted alkylene)-, C(O)NH(substituted or unsubstituted
alkylene), C(O)NH(substituted or unsubstituted
alkenylene)-NHC(O)(substituted or unsubstituted alkylene)-,
--NHC(O)(substituted or unsubstituted alkenylene)-,
--C(O)(substituted or unsubstituted alkenylene)-, and
--NHC(O)(substituted or unsubstituted alkylene)S(substituted or
unsubstituted alkylene)C(O)NH--; and
[0113] T is a substituted or unsubstituted cycloalkyl,
heterocycloalkyl, aryl, or heteroaryl.
[0114] Provided herein are compositions and methods for treating a
disease comprising administering to a subject in need thereof an
effective amount of a FLT-3 modulating compound having the
structure: 39
[0115] wherein:
[0116] each Z is independently C, CR.sub.3, N, NR.sub.3, O, or S,
provided that no more than two Z's are heteroatoms where
[0117] R.sub.3 is H, substituted or unsubstituted alkyl,
substituted or unsubstituted cycloalkyl, substituted or
unsubstituted heteroaryl, or substituted or unsubstituted aryl;
[0118] each R.sub.2 is independently H, substituted or
unsubstituted alkyl, substituted or unsubstituted cycloalkyl,
substituted or unsubstituted heterocyclyl, substituted or
unsubstituted aryl, or substituted or unsubstituted heteroaryl; or
wherein two R.sub.2 groups are linked together by an optionally
substituted alkylene; and
[0119] each R.sub.1 is independently H, halogen, substituted or
unsubstituted alkyl, substituted or unsubstituted alkoxy,
substituted or unsubstituted cycloalkyl, substituted or
unsubstituted heterocyclyl, substituted or unsubstituted aryl,
substituted or unsubstituted heteroaryl, --OR.sub.c --OH,
--OC(O)R.sub.c, --NO.sub.2, --N(R.sub.c).sub.2, --SR.sub.c,
S(O).sub.jR.sub.c where j is 1 or 2, --NR.sub.cC(O)R.sub.c,
--C(O)N(R.sub.c).sub.2, C(O).sub.2R.sub.c, or --C(O)R.sub.c; or two
adjacent R.sub.1's, are taken together to form a substituted or
unsubstituted aryl or heteroaryl, where
[0120] each R.sub.c is independently H, substituted or
unsubstituted alkyl, substituted or unsubstituted cycloalkyl,
substituted or unsubstituted aryl, or substituted or unsubstituted
heteroaryl.
[0121] Provided herein are compositions and methods for treating a
disease comprising administering to a subject in need thereof an
effective amount of a FLT-3 modulating compound having the
structure: 40
[0122] In some embodiments, the protein tyrosine kinase is selected
from the fetus liver kinase ("flk") receptor subfamily, which
includes kinase insert domain-receptor fetal liver kinase-1
(KDR/FLK-1), flk-1R, flk-4 and fins-like tyrosine kinase 1 (flt-1).
In further embodiments, the protein tyrosine kinase is selected
from the fibroblast growth factor ("FGF") receptor subgroup, which
includes the receptors FGFR1, FGFR 2, FGFR3, and FGFR4, and the
ligands, FGF1, FGF2, FGF3, FGF4, FGF5, FGF6, and FGF7. In a still
further embodiment, the protein tyrosine kinase is the tyrosine
kinase growth factor receptor family, c-Met. In some embodiments,
the protein tyrosine kinase is an fins-like tyrosine kinase 3
receptor kinase (FLT-3 kinase).
[0123] The compounds and compositions disclosed herein may be used
for the prevention or treatment of cancers such as stomach,
gastric, bone, ovary, colon, lung, brain, larynx, lymphatic system,
genitourinary tract, ovarian, squamous cell carcinoma, astrocytoma,
Kaposi's sarcoma, glioblastoma, lung cancer, bladder cancer, head
and neck cancer, melanoma, ovarian cancer, prostate cancer, breast
cancer, small-cell lung cancer, leukemia, glioma, colorectal
cancer, genitourinary cancer gastrointestinal cancer, or pancreatic
cancer. In particular, the cancer is acute myelogenous leukemia
(AML), B-precursor cell acute lymphoblastic leukemias,
myelodysplastic leukemias, T-cell acute lymphoblastic leukemias,
and chronic myelogenous leukemias (CMLs).
[0124] Compositions and methods for treating a disease comprising
administering to a subject in need thereof an effective amount of
an fms-like tyrosine kinase 3 (FLT-3) receptor modulating compound
are provided herein. In one embodiment, the disease is cancer. In
other embodiments, the cancer is a malignant tumor, or a
hematologic malignancy such as leukemia and lymphoma. In some
embodiments, the leukemia is acute myelogenous leukemia (AML), a
B-precursor cell acute lymphoblastic leukemia, myelodysplastic
leukemia, T-cell acute lymphoblastic leukemia or chronic
myelogenous leukemia (CML).
[0125] These and other aspects of the present invention will become
evident upon reference to the following detailed description. In
addition, various references are set forth herein which describe in
more detail certain procedures or compositions, and are
incorporated by reference in their entirety.
DISCLOSURE OF THE INVENTION
[0126] To more readily facilitate an understanding of the invention
and its preferred embodiments, the meanings of terms used herein
will become apparent from the context of this specification in view
of common usage of various terms and the explicit definitions of
other terms provided in the glossary below or in the ensuing
description.
[0127] Glossary of Terms
[0128] Unless otherwise stated, the following terms used in this
application, including the specification and claims, have the
definitions given below. It must be noted that, as used in the
specification and the appended claims, the singular forms "a," "an"
and "the" include plural referents unless the context clearly
dictates otherwise. Definition of standard chemistry terms may be
found in reference works, including Carey and Sundberg (1992)
"ADVANCED ORGANIC CHEMISTRY 3.sup.RD ED." Vols. A and B, Plenum
Press, New York. Unless otherwise indicated, conventional methods
of mass spectroscopy, NMR, HPLC, protein chemistry, biochemistry,
recombinant DNA techniques and pharmacology, within the skill of
the art are employed.
[0129] The term "modulator" means a molecule that interacts with a
target either directly or indirectly. The interactions include, but
are not limited to, agonist, antagonist, and the like.
[0130] The term "agonist" means a molecule such as a compound, a
drug, an enzyme activator or a hormone that enhances the activity
of another molecule or the activity of a receptor site etiehr
directly or indirectly.
[0131] The term "antagonist" means a molecule such as a compound, a
drug, an enzyme inhibitor, or a hormone, that diminishes or
prevents the action of another molecule or the activity of a
receptor site either directly or indirectly.
[0132] The terms "effective amount" or "therapeutically effective
amount" refer to a sufficient amount of the agent to provide the
desired biological result. That result can be reduction and/or
alleviation of the signs, symptoms, or causes of a disease, or any
other desired alteration of a biological system. For example, an
"effective amount" for therapeutic use is the amount of the
composition comprising a compound as disclosed herein required to
provide a clinically significant decrease in a disease. An
appropriate "effective" amount in any individual case may be
determined by one of ordinary skill in the art using routine
experimentation.
[0133] As used herein, the terms "treat" or "treatment" are
synonymous with the term "prevent" and are meant to indicate a
postponement of development of diseases, preventing the development
of diseases, and/or reducing severity of such symptoms that will or
are expected to develop. Thus, these terms include ameliorating
existing disease symptoms, preventing additional symptoms,
ameliorating or preventing the underlying metabolic causes of
symptoms, inhibiting the disorder or disease, e.g., arresting the
development of the disorder or disease, relieving the disorder or
disease, causing regression of the disorder or disease, relieving a
condition caused by the disease or disorder, or stopping the
symptoms of the disease or disorder.
[0134] By "pharmaceutically acceptable" or "pharmacologically
acceptable" is meant a material which is not biologically or
otherwise undesirable, i.e., the material may be administered to an
individual without causing any undesirable biological effects or
interacting in a deleterious manner with any of the components of
the composition in which it is contained.
[0135] "Carrier materials" include any commonly used excipients in
pharmaceutics and should be selected on the basis of compatibility
and the release profile properties of the desired dosage form.
Exemplary carrier materials include, e.g., binders, suspending
agents, disintegration agents, filling agents, surfactants,
solubilizers, stabilizers, lubricants, wetting agents, diluents,
and the like. "Pharmaceutically compatible carrier materials" may
comprise, e.g., acacia, gelatin, colloidal silicon dioxide, calcium
glycerophosphate, calcium lactate, maltodextrin, glycerine,
magnesium silicate, sodium caseinate, soy lecithin, sodium
chloride, tricalcium phosphate, dipotassium phosphate, sodium
stearoyl lactylate, carrageenan, monoglyceride, diglyceride,
pregelatinized starch, and the like. See, e.g., Remington: The
Science and Practice of Pharmacy, Nineteenth Ed (Easton, Pa.: Mack
Publishing Company, 1995); Hoover, John E., Remington's
Pharmaceutical Sciences, Mack Publishing Co., Easton, Pa. 1975;
Liberman, H. A. and Lachman, L., Eds., Pharmaceutical Dosage Forms,
Marcel Decker, New York, N.Y., 1980; and Pharmaceutical Dosage
Forms and Drug Delivery Systems, Seventh Ed. (Lippincott Williams
& Wilkins 1999).
[0136] As used herein, the term "subject" encompasses mammals and
non-mammals. Examples of mammals include, but are not limited to,
any member of the Mammalian class: humans, non-human primates such
as chimpanzees, and other apes and monkey species; farm animals
such as cattle, horses, sheep, goats, swine; domestic animals such
as rabbits, dogs, and cats; laboratory animals including rodents,
such as rats, mice and guinea pigs, and the like. Examples of
non-mammals include, but are not limited to, birds, fish and the
like. In one embodiment of the present invention, the mammal is a
human.
[0137] The term "pharmaceutically acceptable salt" of a compound
means a salt that is pharmaceutically acceptable and that possesses
the desired pharmacological activity of the parent compound. Such
salts, for example, include: (1) acid addition salts, formed with
inorganic acids such as hydrochloric acid, hydrobromic acid,
sulfuric acid, nitric acid, phosphoric acid, and the like; or
formed with organic acids such as acetic acid, propionic acid,
hexanoic acid, cyclopentanepropionic acid, glycolic acid, pyruvic
acid, lactic acid, malonic acid, succinic acid, malic acid, maleic
acid, fumaric acid, tartaric acid, citric acid, benzoic acid,
3-(4-hydroxybenzoyl)benzoic acid, cinnamic acid, mandelic acid,
methanesulfonic acid, ethanesulfonic acid, 1,2-ethanedisulfonic
acid, 2-hydroxyethanesulfonic acid, benzenesulfonic acid,
2-naphthalenesulfonic acid,
4-methylbicyclo-[2.2.2]oct-2-ene-1-carboxylic acid, glucoheptonic
acid, 4,4,-methylenebis-(3-hydroxy-2-ene-1-carboxylic acid),
3-phenylpropionic acid, trimethylacetic acid, tertiary butylacetic
acid, lauryl sulfuric acid, gluconic acid, glutamic acid,
hydroxynaphthoic acid, salicylic acid, stearic acid, muconic acid,
and the like; (2) salts formed when an acidic proton present in the
parent compound either is replaced by a metal ion, e.g., an alkali
metal ion, an alkaline earth ion, or an aluminum ion; or
coordinates with an organic base. Acceptable organic bases include
ethanolamine, diethanolamine, triethanolamine, tromethamine,
N-methylglucamine, and the like. Acceptable inorganic bases include
aluminum hydroxide, calcium hydroxide, potassium hydroxide, sodium
carbonate, sodium hydroxide, and the like. It should be understood
that a reference to a pharmaceutically acceptable salt includes the
solvent addition forms or crystal forms thereof, particularly
solvates or polymorphs. Solvates contain either stoichiometric or
non-stoichiometric amounts of a solvent, and are often formed
during the process of crystallization. Hydrates are formed when the
solvent is water, or alcoholates are formed when the solvent is
alcohol. Polymorphs include the different crystal packing
arrangements of the same elemental composition of a compound.
Polymorphs usually have different X-ray diffraction patterns,
infrared spectra, melting points, density, hardness, crystal shape,
optical and electrical properties, stability, and solubility.
Various factors such as the recrystallization solvent, rate of
crystallization, and storage temperature may cause a single crystal
form to dominate.
[0138] As used herein, the term "biological sample" is broadly
defined to include any cell, tissue, organ or multicellular
organism. A biological sample can be derived, for example, from
cell or tissue cultures in vitro. Alternatively, a biological
sample can be derived from a living organism or from a population
of single cell organisms.
[0139] As used herein, the term "linker" means any divalent linking
moiety used to connect, join, or attach two chemical groups. For
example, linkers may be used to join two cyclic groups, such as to
join two aryl groups (e.g., phenyl), an aryl group to a cycloalkyl
group, an aryl group to a heterocyclyl group, a cycloalkyl group to
a cycloalkyl group, a cycloalkyl group to a heterocyclyl group, and
the like. Representative linkers include, but are not limited to, a
covalent bond, -(substituted or unsubstituted alkylene)-,
-(substituted or unsubstituted alkenylene)-, -(substituted or
unsubstituted alkynylene)-, -(substituted or unsubstituted
cycloalkylene)-, -(substituted or unsubstituted heterocyclylene)-,
-(substituted or unsubstituted arylene)-, and -(substituted or
unsubstituted heteroarylene)-. Exemplary linkers also include
--O--, --S--, --S(O)--, --S(O).sub.2--, --S(O).sub.3--, --C(O)--,
--NH--, --N.dbd., --N.dbd.N--, .dbd.N--N.dbd., --C(O)NH--,
--S(O)NH--, and the like. Additional examples of linkers include
--O(substituted or unsubstituted alkylene)-, --N(substituted or
unsubstituted alkylene)-, --NHC(O)(substituted or unsubstituted
alkylene)-, --C(O)(substituted or unsubstituted alkenylene)-,
--NHC(O)(substituted or unsubstituted alkylene)S(substituted or
unsubstituted alkylene)C(O)NH--, --NHC(O)(substituted or
unsubstituted alkenylene)-, and the like. Linkers, as represented
herein, embrace divalent moieties in any chemically feasible
directionality. For example, compounds comprising a linker-C(O)NH--
which attaches two aryl groups, Ar.sub.1 to Ar.sub.2, include
Ar.sub.1-C(O)NH-Ar.sub.2 as well as Ar.sub.1-NHC(O)-Ar.sub.2.
[0140] As used herein, the term "halogen" includes fluorine,
chlorine, bromine, and iodine.
[0141] As used herein, "alkyl" means a straight chain or branched,
saturated or unsaturated chain having from 1 to 10 carbon atoms.
Representative saturated alkyl groups include, but are not limited
to, methyl, ethyl, n-propyl, isopropyl, 2-methyl-1-propyl,
2-methyl-2-propyl, 2-methyl-1-butyl, 3-methyl-1-butyl,
2-methyl-3-butyl, 2,2-dimethyl-1-propyl, 2-methyl-1-pentyl,
3-methyl-1-pentyl, 4-methyl-1-pentyl, 2-methyl-2-pentyl,
3-methyl-2-pentyl, 4-methyl-2-pentyl, 2,2-dimethyl-1-butyl,
3,3-dimethyl-1-butyl, 2-ethyl-1-butyl, butyl, isobutyl, t-butyl,
n-pentyl, isopentyl, neopentyl, and n-hexyl, and longer alkyl
groups, such as heptyl, and octyl. An alkyl group can be
unsubstituted or substituted. Unsaturated alkyl groups include
alkenyl groups and alkynyl groups, discussed below. Alkyl groups
containing three or more carbon atoms may be straight, branched or
cyclized.
[0142] As used herein, "lower alkyl" means an alkyl having from 1
to 5 carbon atoms.
[0143] As used herein, an "alkenyl group" includes a monovalent
unbranched or branched hydrocarbon chain having one or more double
bonds therein. The double bond of an alkenyl group can be
unconjugated or conjugated to another unsaturated group. Suitable
alkenyl groups include, but are not limited to, (C.sub.2-C.sub.8)
alkenyl groups, such as vinyl, allyl, butenyl, pentenyl, hexenyl,
butadienyl, pentadienyl, hexadienyl, 2-ethylhexenyl,
2-propyl-2-butenyl, 4-(2-methyl-3-butene)-pentenyl. An alkenyl
group can be unsubstituted or substituted.
[0144] As used herein, "alkynyl group" includes a monovalent
unbranched or branched hydrocarbon chain having one or more triple
bonds therein. The triple bond of an alkynyl group can be
unconjugated or conjugated to another unsaturated group. Suitable
alkynyl groups include, but are not limited to, (C.sub.2-C.sub.6)
alkynyl groups, such as ethynyl, propynyl, butynyl, pentynyl,
hexynyl, methylpropynyl, 4-methyl-1-butynyl, 4-propyl-2-pentynyl,
and 4-butyl-2-hexynyl. An alkynyl group can be unsubstituted or
substituted.
[0145] The terms "trifluoromethyl," "sulfonyl," and "carboxyl"
include CF.sub.3, SO.sub.3H, and CO.sub.2H, respectively.
[0146] The term "alkoxy" as used herein includes --O-(alkyl),
wherein alkyl is defined above.
[0147] As used herein, "alkoxyalkoxy" includes
--O-(alkyl)-O-(alkyl), wherein each "alkyl" is independently an
alkyl group defined above.
[0148] As used herein, "alkoxycarbonyl" includes-C(O)O-(alkyl),
wherein alkyl is defined above.
[0149] As used herein, "alkoxycarbonylalkyl" includes
-(alkyl)-C(O)O-(alkyl), wherein alkyl is defined above.
[0150] As used herein, "alkoxyalkyl" means -(alkyl)-O-(alkyl),
wherein each "alkyl" is independently an alkyl group defined
above.
[0151] As used herein, the term "aryl" (Ar) refers to a monocyclic,
or fused or spiro polycyclic, aromatic carbocycle (ring structure
having ring atoms that are all carbon) having from 3 to 12 ring
atoms per ring. Illustrative examples of aryl groups include the
following moieties: 41
[0152] As used herein, the term "heteroaryl" (heteroAr) refers to a
monocyclic, or fused or Spiro polycyclic, aromatic heterocycle
(ring structure having ring atoms selected from carbon atoms as
well as nitrogen, oxygen, and sulfur heteroatoms) having from 3 to
12 ring atoms per ring. Illustrative examples of aryl groups
include the following moieties: 42
[0153] As used herein, the term "cycloalkyl" refers to a saturated
or partially saturated, monocyclic or fused or spiro polycyclic,
carbocycle having from 3 to 12 ring atoms per ring. Illustrative
examples of cycloalkyl groups include the following moieties:
43
[0154] As used herein, the term "heterocycloalkyl" refers to a
monocyclic, or fused or spiro polycyclic, ring structure that is
saturated or partially saturated and has from 3 to 12 ring atoms
per ring selected from C atoms and N, O, and S heteroatoms.
Illustrative examples of heterocycloalkyl groups include: 44
[0155] As used herein, "aryloxy" includes --O-aryl group, wherein
aryl is as defined above. An aryloxy group can be unsubstituted or
substituted.
[0156] As used herein, "arylalkyl" includes -(alkyl)-(aryl),
wherein alkyl and aryl are defined above.
[0157] As used herein, "arylalkyloxy" includes --O-(alkyl)-(aryl),
wherein alkyl and aryl are defined above.
[0158] As used herein, "cycloalkyl" includes a monocyclic or
polycyclic saturated ring comprising carbon and hydrogen atoms and
having no carbon-carbon multiple bonds. Examples of cycloalkyl
groups include, but are not limited to, (C.sub.3-C.sub.7)cycloalkyl
groups, such as cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl,
and cycloheptyl, and saturated cyclic and bicyclic terpenes. A
cycloalkyl group can be unsubstituted or substituted. Preferably,
the cycloalkyl group is a monocyclic ring or bicyclic ring.
[0159] As used herein, "cycloalkyloxy" includes --O-(cycloalkyl),
wherein cycloalkyl is defined above.
[0160] As used herein, "cycloalkylalkyloxy" includes
--O-(alkyl)-(cycloalkyl), wherein cycloalkyl and alkyl are defined
above.
[0161] As used herein, the term "alkylidene" includes the divalent
radical --C.sub.nH.sub.2n--, wherein n is an integer from 1 to 8,
such as --CH.sub.2--, --CH.sub.2CH.sub.2--,
--CH.sub.2--CH.sub.2--CH.sub.2--,
--CH.sub.2CH.sub.2CH.sub.2CH.sub.2--,
--CH.sub.2CH.sub.2CH.sub.2CH.sub.2C- H.sub.2--, and the like,
unsubstituted or substituted with one or more alkyl groups.
[0162] As used herein, "heteroatom-containing alkylidene" includes
an alkylidene wherein at least one carbon atom is replaced by a
heteroatom selected from nitrogen, oxygen, or sulfur, such as
--CH.sub.2CH.sub.2OCH.sub.2CH.sub.2--, and the like, unsubstituted
or substituted with one or more alkyl groups.
[0163] As used herein, "aminoalkoxy" includes --O-(alkyl)-NH.sub.2,
wherein alkyl is defined above.
[0164] As used herein, "mono-alkylamino" includes --NH(alkyl),
wherein alkyl is defined above.
[0165] As used herein, "di-alkylamino" includes --N(alkyl)(alkyl),
wherein each "alkyl" is independently an alkyl group defined
above.
[0166] As used herein, "mono-alkylaminoalkoxy" includes
--O-(alkyl)-NH(alkyl), wherein each "alkyl" is independently an
alkyl group defined above.
[0167] As used herein, "di-alkylaminoalkoxy" includes
--O-(alkyl)N(alkyl)(alkyl), wherein each "alkyl" is independently
an alkyl group defined above.
[0168] As used herein, "arylamino" includes --NH(aryl), wherein
aryl is defined above.
[0169] As used herein, "arylalkylamino" includes
--NH-(alkyl)-(aryl), wherein alkyl and aryl are defined above.
[0170] As used herein, "alkylamino" includes --NH(alkyl), wherein
alkyl is defined above.
[0171] As used herein, "cycloalkylamino" includes
--NH-(cycloalkyl), wherein cyclohexyl is defined above.
[0172] As used herein, "cycloalkylalkylamino" includes
--NH-(alkyl)-(cycloalkyl), wherein alkyl and cycloalkyl are defined
above.
[0173] As used herein, "aminoalkyl" includes -(alkyl)-NH.sub.2,
wherein alkyl is defined above.
[0174] As used herein, "mono-alkylaminoalkyl" includes
-(alkyl)-NH(alkyl), wherein each "alkyl" is independently an alkyl
group defined above.
[0175] As used herein, "di-alkylaminoalkyl" includes
-(alkyl)-N(alkyl)(alkyl), wherein each "alkyl" is independently an,
alkyl group defined above.
[0176] The term "whole integer" is intended to include whole
numbers. For example, a whole integer from 0 to 4 would include 0,
1, 2, 3, and 4.
[0177] Sulfonyl refers to the presence of a sulfur atom, which is
optionally linked to another moiety such as an aliphatic group, an
aromatic group, an aryl group, an alicyclic group, or a
heterocyclic group. Aryl or alkyl sulfonyl moieties have the
formula --SO.sub.2R.sub.d, and alkoxy moieties have the formula
--O--R.sub.d, wherein R.sub.d is alkyl, as defined above, or is
aryl wherein aryl is phenyl, optionally substituted with 1-3
substituents independently selected from halo (fluoro, chloro,
bromo or iodo), lower alkyl (1-6C) and lower alkoxy (1-6C).
[0178] As used herein, the term "substituted" means that the
specified group or moiety bears one or more suitable
substituents.
[0179] As used herein, the term "unsubstituted" means that the
specified group bears no substituents.
[0180] As used herein, the term "optionally substituted" means that
the specified group is unsubstituted or substituted by one or more
substituents.
[0181] Molecular embodiments of the present invention may possess
one or more chiral centers and each center may exist in the R or S
configuration. The present invention includes all diastereomeric,
enantiomeric, and epimeric forms as well as the appropriate
mixtures thereof. Stereoisomers may be obtained, if desired, by
methods known in the art as, for example, the separation of
stereoisomers by chiral chromatographic columns. Additionally, the
compounds of the present invention may exist as geometric isomers.
The present invention includes all cis, trans, syn, anti, entgegen
(E), and zusammen (Z) isomers as well as the appropriate mixtures
thereof.
[0182] Certain functional groups contained within the compounds of
the present invention can be substituted for bioisosteric groups,
that is, groups which have similar spatial or electronic
requirements to the parent group, but exhibit differing or improved
physicochemical or other properties. Suitable examples are well
known to those of skill in the art, and include, but are not
limited to moieties described in Patini et al., Chem, Rev, 1996,
96, 3147-3176 and references cited therein.
[0183] In addition, the compounds of the present invention can
exist in unsolvated as well as solvated forms with pharmaceutically
acceptable solvents such as water, ethanol, and the like. In
general, the solvated forms are considered equivalent to the
unsolvated forms for the purposes of the present invention.
[0184] To more readily facilitate an understanding of the invention
and its preferred embodiments, the meanings of terms used herein
will become apparent from the context of this specification in view
of common usage of various terms and the explicit definitions of
other terms provided in the glossary below or in the ensuing
description.
[0185] Compounds
[0186] In one aspect, the present invention is directed to
compounds, compositions, and methods for treating conditions
associated with abnormal kinase activity. In one embodiment,
compounds useful in the invention are derivatives of isoxazoles,
pyrazoles and isothiazoles. When the compounds of the invention
contain one or more chiral centers, the invention includes
optically pure forms as well as mixtures of stereoisomers or
enantiomers.
[0187] Thus, the invention provides methods for modulating various
kinases by providing an effective amount of a compound of the
formulas described herein.
[0188] Salts of the compounds may be used for therapeutic and
prophylactic purposes, where the salt is preferably a
pharmaceutically acceptable salt. Examples of pharmaceutically
acceptable salts include those derived from mineral acids, such as
hydrochloric, hydrobromic, phosphoric, metaphosphoric, nitric and
sulphuric acids, and organic acids, such as tartaric, acetic,
trifluoroacetic, citric, malic, lactic, fumaric, benzoic, glycolic,
gluconic, succinic and methanesulphonic and arylsulphonic, for
example Q-toluenesulphonic, acids.
[0189] A "prodrug" refers to a drug or compound in which the
pharmacological action results from conversion by metabolic
processes within the body. Prodrugs are generally drug precursors
that, following administration to a subject and subsequent
absorption, are converted to an active, or a more active species
via some process, such as conversion by a metabolic pathway. Some
prodrugs have a chemical group present on the prodrug that renders
it less active and/or confers solubility or some other property to
the drug. Once the chemical group has been cleaved and/or modified
from the prodrug the active drug is generated. Prodrugs may be
designed as reversible drug derivatives, for use as modifiers to
enhance drug transport to site-specific tissues. Additionally,
prodrugs can increase the effective water solubility of the
therapeutic compound for targeting to regions where water is the
principal solvent. See, e.g., Fedorak et al., Am. J. Physiol., 269:
G210-218 (1995); McLoed et al., Gastroenterol, 106: 405-413 (1994);
Hochhaus et al., Biomed. Chrom., 6: 283-286 (1992); J. Larsen and
H. Bundgaard, Int. J. Pharmaceutics, 37, 87 (1987); J. Larsen et
al., Int. J. Pharmaceutics, 47, 103 (1988); Sinkula et al., J.
Pharm. Sci., 64: 181-210 (1975); T. Higuchi and V. Stella,
Pro-drugs as Novel Delivery Systems, Vol. 14 of the A.C.S.
Symposium Series; and Edward B. Roche, Bioreversible Carriers in
Drug Design, American Pharmaceutical Association and Pergamon
Press, 1987. Prodrug forms of the above described compounds,
wherein the prodrug is metabolized in vivo to produce a derivative
as set forth above are included within the scope of the claims.
Indeed, some of the above-described derivatives may be a prodrug
for another derivative or active compound. The invention further
provides for the optical isomers of the compounds disclosed herein,
especially those resulting from the chiral carbon atoms in the
molecule. In additional embodiments of the invention, mixtures of
enantiomers and/or diastereoisomers, resulting from a single
preparative step, combination, or interconversion may also be
useful for the applications described herein.
[0190] In another aspect, compositions containing the above
described analogs and derivatives are provided. Preferably, the
compositions are formulated to be suitable for pharmaceutical or
clinical use by the inclusion of appropriate carriers or
excipients.
[0191] Groups such as carbonyl, carboxyl, alkoxy, amino, and cyano
groups, etc., as shown in the formula above, need not be directly
bound to the para position; they may be included elsewhere in the
alkyl, alkenyl or alkynyl substituent. Thus, also acceptable
substituents are the following representative forms:
[0192] --CH.sub.2NHCH.sub.3; --CH.sub.2OCH.sub.3;
--CH.sub.2SCH.sub.3; --NHCH.sub.3; --CH.sub.2CH.sub.3;
--OCH.sub.2CH.sub.3; --SCH.sub.2CH.sub.2CH.sub.3;
--CH.dbd.CHCH.sub.2NH.sub.2; --CH.sub.2CH.sub.2OH; 45
[0193] --CH.sub.2CH.sub.2CH.sub.2SH; --CH.sub.2OC(O)CH.sub.3;
--CH.sub.2NHC(O)CH.sub.2C(O)CH.sub.3;
--NHC(O)CH.sub.2CH.sub.2CH.sub.3 each of which may further be
substituted with a cycloalkyl, heterocycloalkyl, aryl or heteroaryl
group.
[0194] It will also be evident that these substituents include, for
example, trifluoromethyl, difluoromethyl and fluoromethyl (alkyl
substituted by halo) and trifluoromethoxy, difluoromethoxy and
fluoromethoxy (alkyl where one carbon is replaced by 0 and is
further substituted by halo).
[0195] Compounds of the invention which contain carboxyl groups or
which contain amino groups may be supplied in the forms of their
pharmaceutically acceptable salts. Pharmaceutically acceptable
salts of carboxylic acids include inorganic salts such as salts of
sodium, potassium, calcium, magnesium and the like or salts formed
with organic bases such as caffeine. Salts of amines are acid
addition salts formed from inorganic acids such as hydrochloric,
sulfuric, phosphoric acids or may be salts of organic acids such as
acetates, maleates, propionates, and the like.
[0196] The invention also provides prodrug forms of the compounds
described herein, wherein the prodrug is metabolized in vivo to
produce a derivative as set forth above. Indeed, some of the above
described derivatives may be a prodrug for another derivative or
active compound. The invention further provides for the optical
isomers of the compounds disclosed herein, especially those
resulting from the chiral carbon atoms in the molecule. In
additional embodiments of the invention, mixtures of enantiomers
and/or diastereoisomers, resulting from a single preparative step,
combination, or interconversion are provided.
[0197] In another aspect of the invention, compositions containing
the above described analogs and derivatives are provided.
Preferably, the compositions are formulated to be suitable for
pharmaceutical or clinical use by the inclusion of appropriate
carriers or excipients.
[0198] In yet another aspect of the invention, pharmaceutical
formulations are provided comprising at least one compound
described above, or a pharmaceutically acceptable salt or solvate
thereof, together with one or more pharmaceutically acceptable
carriers, diluents or excipients.
[0199] The compounds of the invention, especially when used in the
invention methods and compositions, may be "conjugated"--that is
they may be coupled to additional moieties that do not destroy
their ability to modulate kinases. For example, the compounds might
be coupled to a label such as a radioactive label, a fluorescent
label and the like, or may be coupled to targeting agents such as
antibodies or fragments, or to fragments to aid purification such
as FLAG or a histidine tag. The compounds may also be coupled to
specific binding partners such as biotin for use in assay
procedures or to moieties that alter their biological half-lives
such as polyethylene glycol. Thus, the methods of the invention
employ the invention compounds per se as well as conjugates
thereof.
[0200] Synthesis of Compounds
[0201] Compounds of the present invention may be synthesized using
standard synthetic techniques known to those of skill in the art or
using methods known in the art in combination with methods
described herein. See, e.g., March, ADVANCED ORGANIC CHEMISTRY
4.sup.th Ed., (Wiley 1992); Carey and Sundberg, ADVANCED ORGANIC
CHEMISTRY 3.sup.rd Ed., Vols. A and B (Plenum 1992), and Green and
Wuts, PROTECTIVE GROUPS IN ORGANIC SYNTHESIS 2.sup.nd Ed. (Wiley
1991). General methods for the preparation of compound as disclosed
herein may be derived from known reactions in the field, and the
reactions may be modified by the use of appropriate reagents and
conditions, as would be recognized by the skilled person, for the
introduction of the various moieties found in the formulae as
provided herein.
[0202] The compounds of the invention are synthesized by methods
well known in the art. The compounds of the invention are ureas or
cyclic forms thereof and can be synthesized using generally known
procedures for urea synthesis.
[0203] In one group of methods, an amine is reacted with an
isocyanate in an aprotic solvent. Typically, in some embodiments, a
molar excess of the amine is used in the presence of an aprotic
solvent and the reaction is conducted at room temperature. The
reaction mixture is then poured into water and precipitated with
salt to recover the crude product which is then purified according
to standard methods.
[0204] In alternative methods, the ureas are formed from two
separate amine reactants in the presence of a condensing agent such
as 1,1,carbonyldiimidazole (CDI) in the presence of an inert
nonpolar solvent such as dichloromethane. One of the amines is
first added to a solution of CDI in solvent under cooling
conditions and then stirred at room temperature with the other
amine. After removal of solvent, the crude product can be purified
using standard procedures.
[0205] In still another method, one of the amines is added in an
aprotic solvent to a solution of triphosgene and then treated with
the other amine reactant dissolved in an inert solvent in the
presence of base such as triethylamine. After reaction at room
temperature, the mixture may be diluted with, for example,
ethylacetate and washed with water and brine, dried and
purified.
[0206] In still another method, one of the amine components is
treated with 4-nitrophenylchloroformate in the presence of mild
base in a solvent such as N-methylpyrrolidone (NMP). The other
amine is then added and the reaction mixture heated, then cooled,
poured into water, extracted into chloroform and further
purified.
[0207] Alternatively, the urea may be formed by the reaction of an
amine with the counterpart halo acylamine which is formed from the
parent amine by treatment with phosgene and base in an inert
solvent such as methylene dichloride or by reacting an amine with
its counterpart amine with an acyl amine containing an alternate
leaving group formed by reaction of that amine with
4-nitrophenylchloroformate in the presence of an amine base and in
an inert solvent.
[0208] Details of these methods can be found in Matsuno et al. J.
Med. Chem. 45: 3057-66 (2002); Matsuno et al. J. Med. Chem. 45:
4513-23 (2002); and and Pandley et al., J. Med. Chem. 45: 3772-93
(2002).
[0209] Cyclized forms of the ureas may be obtained by treating the
formed urea with dibromo derivatives of the bridge, typically in
the presence of a strong base and in an inert aprotic polar
solvent.
[0210] The ureas may be converted to thioureas by treating with
Lawesson's reagent in the presence of toluene.
[0211] For compounds having the moiety Ar.sup.1-L-Ar.sup.2 is
obtained by first protecting the amino group of p-hydroxy aniline
destined to become Ar.sup.1 with a protecting agent such as Boc and
then coupling the hydroxy group of Ar.sup.1 to an aryl alkyl
halide. This coupling is conducted in the presence of strong base
and in an aprotic solvent. After deprotection, the urea is formed
by reaction with the isoxazole isocyanate. These techniques are
exemplified below.
[0212] Selected examples of covalent linkages and precursor
functional groups which yield them are given in the Table entitled
"Examples of Covalent Linkages and Precursors Thereof." Precursor
functional groups are shown as electrophilic groups and
nucleophilic groups. The functional group on the organic substance
may be attached directly, or attached via any useful spacer or
linker as defined below.
1TABLE 1 Examples of Covalent Linkages and Precursors Thereof
Covalent Linkage Product Electrophile Nucleophile Carboxamides
Activated esters amines/anilines Carboxamides acyl azides
amines/anilines Carboxamides acyl halides amines/anilines Esters
acyl halides alcohols/phenols Esters acyl nitriles alcohols/phenols
Carboxamides acyl nitriles amines/anilines Imines Aldehydes
amines/anilines Hydrazones aldehydes or ketones Hydrazines Oximes
aldehydes or ketones Hydroxylamines Alkyl amines alkyl halides
amines/anilines Esters alkyl halides carboxylic acids Thioethers
alkyl halides Thiols Ethers alkyl halides alcohols/phenols
Thioethers alkyl sulfonates Thiols Esters alkyl sulfonates
carboxylic acids Ethers alkyl sulfonates alcohols/phenols Esters
Anhydrides alcohols/phenols Carboxamides Anhydrides amines/anilines
Thiophenols aryl halides Thiols Aryl amines aryl halides Amines
Thioethers Azindines Thiols Boronate esters Boronates Glycols
Carboxamides carboxylic acids amines/anilines Esters carboxylic
acids Alcohols hydrazines Hydrazides carboxylic acids N-acylureas
or Anhydrides carbodiimides carboxylic acids Esters diazoalkanes
carboxylic acids Thioethers Epoxides Thiols Thioethers
haloacetamides Thiols Ammotriazines halotriazines amines/anilines
Triazinyl ethers halotriazines alcohols/phenols Amidines imido
esters amines/anilines Ureas Isocyanates amines/anilines Urethanes
Isocyanates alcohols/phenols Thioureas isothiocyanates
amines/anilines Thioethers Maleimides Thiols Phosphite esters
phosphoramidites Alcohols Silyl ethers silyl halides Alcohols Alkyl
amines sulfonate esters amines/anilines Thioethers sulfonate esters
Thiols Esters sulfonate esters carboxylic acids Ethers sulfonate
esters Alcohols Sulfonamides sulfonyl halides amines/anilines
Sulfonate esters sulfonyl halides phenols/alcohols
[0213] In general, carbon electrophiles are susceptible to attack
by complementary nucleophiles, including carbon nucleophiles,
wherein an attacking nucleophile brings an electron pair to the
carbon electrophile in order to form a new bond between the
nucleophile and the carbon electrophile.
[0214] Suitable carbon nucleophiles include, but are not limited to
alkyl, alkenyl, aryl and alkynyl Grignard, organolithium,
organozinc, alkyl-, alkenyl, aryl- and alkynyl-tin reagents
(organostannanes), alkyl-, alkenyl-, aryl- and alkynyl-borane
reagents (organoboranes and organoboronates); these carbon
nucleophiles have the advantage of being kinetically stable in
water or polar organic solvents. Other carbon nucleophiles include
phosphorus ylids, enol and enolate reagents; these carbon
nucleophiles have the advantage of being relatively easy to
generate from precursors well known to those skilled in the art of
synthetic organic chemistry. Carbon nucleophiles, when used in
conjunction with carbon electrophiles, engender new carbon-carbon
bonds between the carbon nucleophile and carbon electrophile.
[0215] Non-carbon nucleophiles suitable for coupling to carbon
electrophiles include but are not limited to primary and secondary
amines, thiols, thiolates, and thioethers, alcohols, alkoxides,
azides, semicarbazides, and the like. These non-carbon
nucleophiles, when used in conjunction with carbon electrophiles,
typically generate heteroatom linkages (C--X--C), wherein X is a
hetereoatom, e.g, oxygen or nitrogen.
[0216] The term "protecting group" refers to chemical moieties that
block some or all reactive moieties and prevent such groups from
participating in chemical reactions until the protective group is
removed. It is preferred that each protective group be removable by
a different means. Protective groups that are cleaved under totally
disparate reaction conditions fulfill the requirement of
differential removal. Protective groups can be removed by acid,
base, and hydrogenolysis. Groups such as trityl, dimethoxytrityl,
acetal and t-butyldimethylsilyl are acid labile and may be used to
protect carboxy and hydroxy reactive moieties in the presence of
amino groups protected with Cbz groups, which are removable by
hydrogenolysis, and Fmoc groups, which are base labile. Carboxylic
acid and hydroxy reactive moieties may be blocked with base labile
groups such as, without limitation, methyl, ethyl, and acetyl in
the presence of amines blocked with acid labile groups such as
t-butyl carbamate or with carbamates that are both acid and base
stable but hydrolytically removable.
[0217] Carboxylic acid and hydroxy reactive moieties may also be
blocked with hydrolytically removable protective groups such as the
benzyl group, while amine groups capable of hydrogen bonding with
acids may be blocked with base labile groups such as Fmoc.
Carboxylic acid reactive moieties may be protected by conversion to
simple ester derivatives as exemplified herein, or they may be
blocked with oxidatively-removable protective groups such as
2,4-dimethoxybenzyl, while co-existing amino groups may be blocked
with fluoride labile silyl carbamates.
[0218] Allyl blocking groups are useful in then presence of acid-
and base-protecting groups since the former are stable and can be
subsequently removed by metal or pi-acid catalysts. For example, an
allyl-blocked carboxylic acid can be deprotected with a
Pd.sub.0-catalyzed reaction in the presence of acid labile t-butyl
carbamate or base-labile acetate amine protecting groups. Yet
another form of protecting group is a resin to which a compound or
intermediate may be attached. As long as the residue is attached to
the resin, that functional group is blocked and cannot react. Once
released from the resin, the functional group is available to
react.
[0219] Typically blocking/protecting groups may be selected from:
46
[0220] Other protecting groups are described in Greene and Wuts,
Protective Groups in Organic Synthesis, 3rd Ed., John Wiley &
Sons, New York, N.Y., 1999, which is incorporated herein by
reference in its entirety.
[0221] Biological Activity
[0222] Protein kinases (PKs) play a role in signal transduction
pathways regulating a number of cellular functions, such as cell
growth, differentiation, and cell death. PKs are enzymes that
catalyze the phosphorylation of hydroxy groups on tyrosine, serine
and threonine residues of proteins. Abnormal PK activity has been
related to disorders ranging from relatively non life threatening
diseases such as psoriasis to extremely virulent diseases such as
glioblastoma (brain cancer). In addition, a variety of tumor types
have dysfunctional growth factor receptor tyrosine kinases,
resulting in inappropriate mitogenic signaling. Protein kinases are
believed to be involved in many different cellular signal
transduction pathways. In particular, protein tyrosine kinases
(PTK) are attractive targets in the search for therapeutic agents,
not only for cancer, but also against many other diseases. Blocking
or regulating the kinase phosphorylation process in a signaling
cascade may help treat conditions such as cancer or inflammatory
processes.
[0223] Protein tyrosine kinases are a family of tightly regulated
enzymes, and the aberrant activation of various members of the
family is one of the hallmarks of cancer. The protein-tyrosine
kinase family includes Bcr-Abl tyrosine kinase, and can be divided
into subgroups that have similar structural organization and
sequence similarity within the kinase domain. The members of the
type III group of receptor tyrosine kinases include the
platelet-derived growth factor (PDGF) receptors (PDGF receptors
.alpha. and .beta.), colony-stimulating factor (CSF-1) receptor
(CSF-1R, c-Fms), FLT-3, and stem cell or steel factor receptor
(c-kit).
[0224] The compounds, compositions, and methods provided herein are
useful to modulate the activity of kinases including, but not
limited to, ERBB2, ABL1, AURKA, CDK2, EGFR, FGFR1, LCK, MAPK14,
PDGFR, KDR, ABL1, BRAF, ERBB4, FLT3, KIT, and RAF1. In some
embodiments, the compositions and methods provided herein modulate
the activity of a mutant kinase.
[0225] Inhibition by the compounds provided herein can be
determined using any suitable assay. In one embodiment, inhibition
is determined in vitro. In a specific embodiment, inhibition is
assessed by phosphorylation assays. Any suitable phosphorylation
assay can be employed. For example, membrane autophosphorylation
assays, receptor autophosphorylation assays in intact cells, and
ELISA's can be employed. See, e.g., Gazit, et al., J. Med. Chem.
(1996) 39: 2170-2177, Chapter 18 in CURRENT PROTOCOLS IN MOLECULAR
BIOLOGY (Ausubel, et al., eds. 2001). Cells useful in such assays
include cells with wildtype or mutated forms. In one embodiment,
the wildtype is a kinase that is not constitutively active, but is
activated with upon dimerization. For example, the mutant FLT3
kinase is constitutively active via internal tandem duplication
mutations or point mutations in the activation domain. Suitable
cells include those derived through cell culture from patient
samples as well as cells derived using routine molecular biology
techniques, e.g., retroviral transduction, transfection,
mutagenesis, etc. Exemplary cells include Ba/F3 or 32Dc13 cells
transduced with, e.g., MSCV retroviral constructs FLT3-ITD (Kelly
et al., 2002); Molm-13 and Molm14 cell line (Fujisaki Cell Center,
Okayama, Japan); HL60 (AML-M3), AML193 (AML-M5), KG-1, KG-la,
CRL-1873, CRL-9591, and THP-1 (American Tissue Culture Collection,
Bethesda, Md.); or any suitable cell line derived from a patient
with a hematopoietic malignancy.
[0226] In some embodiments, the compounds described herein
significantly inhibit receptor tyrosine kinases. A significant
inhibition of a receptor tyrosine kinase activity refers to an
IC.sub.50 of less than or equal to 100 .mu.M. Preferably, the
compound can inhibit activity with an IC.sub.50 of less than or
equal to 50 .mu.M, more preferably less than or equal to 10 .mu.M,
more preferably less than 1 .mu.M, or less than 100 nM, most
preferably less than 50 nM. Lower IC.sub.50's are preferred because
the IC.sub.50 provides an indication as to the in vivo
effectiveness of the compound. Other factors known in the art, such
as compound half-life, biodistribution, and toxicity should also be
considered for therapeutic uses. Such factors may enable a compound
with a lower IC.sub.50 to have greater in vivo efficacy than a
compound having a higher IC.sub.50. Preferably, a compound that
inhibits activity is administered at a dose where the effective
tyrosine phosphorylation, i.e., IC.sub.50, is less than its
cytotoxic effects, LD.sub.50.
[0227] In some embodiments, the compounds selectively inhibit one
or more kinases. Selective inhibition of a kinase, such as FLT3,
p38 kinase, STK10, MKNK2, Bcr-Abl, c-kit, or PDGFR, is achieved by
inhibiting activity of one kinase, while having an insignificant
effect on other members of the superfamily.
[0228] FLT-3
[0229] FLT3 kinase is a tyrosine kinase receptor involved in the
regulation and stimulation of cellular proliferation. See e.g.,
Gilliland et al., Blood 100: 1532-42 (2002). The FLT3 kinase is a
member of the class III receptor tyrosine kinase (RTKIII) receptor
family and belongs to the same subfamily of tyrosine kinases as
c-kit, c-fms, and the platelet-derived growth factor .alpha. and
.beta. receptors. See e.g., Lyman et al., FLT3 Ligand in THE
CYTOKINE HANDBOOK 989 (Thomson et al., eds. 4th Ed.) (2003). The
FLT3 kinase has five immunoglobulin-like domains in its
extracellular region as well as an insert region of 75-100 amino
acids in the middle of its cytoplasmic domain. FLT3 kinase is
activated upon the binding of the FLT3 ligand, which causes
receptor dimerization. Dimerization of the FLT3 kinase by FLT3
ligand activates the intracellular kinase activity as well as a
cascade of downstream substrates including Stat5, Ras,
phosphatidylinositol-3-kinase (PI3K), PLC.gamma., Erk2, Akt, MAPK,
SHC, SHP2, and SHIP. See e.g., Rosnet et al., Acta Haematol. 95:
218 (1996); Hayakawa et al., Oncogene 19: 624 (2000); Mizuki et
al., Blood 96: 3907 (2000); and Gilliand et al., Curr. Opin.
Hematol. 9: 274-81 (2002). Both membrane-bound and soluble FLT3
ligand bind, dimerize, and subsequently activate the FLT3
kinase.
[0230] In normal cells, immature hematopoietic cells, typically
CD34+ cells, placenta, gonads, and brain express FLT3 kinase. See,
e.g., Rosnet, et al., Blood 82: 1110-19 (1993); Small et al., Proc.
Natl. Acad. Sci. U.S.A. 91: 459-63 (1994); and Rosnet et al.,
Leukemia 10: 238-48 (1996). However, efficient stimulation of
proliferation via FLT3 kinase typically requires other
hematopoietic growth factors or interleukins. FLT3 kinase also
plays a critical role in immune function through its regulation of
dendritic cell proliferation and differentiation. See e.g., McKenna
et al., Blood 95: 3489-97 (2000).
[0231] Numerous hematologic malignancies express FLT3 kinase, the
most prominent of which is AML. See e.g., Yokota et al., Leukemia
11: 1605-09 (1997). Other FLT3 expressing malignancies include
B-precursor cell acute lymphoblastic leukemias, myelodysplastic
leukemias, T-cell acute lymphoblastic leukemias, and chronic
myelogenous leukemias. See e.g., Rasko et al., Leukemia 9: 2058-66
(1995).
[0232] FLT3 kinase mutations associated with hematologic
malignancies are activating mutations. In other words, the FLT3
kinase is constitutively activated without the need for binding and
dimerization by FLT3 ligand, and therefore stimulates the cell to
grow continuously.
[0233] Several studies have identified inhibitors of FLT3 kinase
activity that also inhibit the kinase activity of related
receptors, e.g., VEGF receptor (VEGFR), PDGF receptor (PDGFR), and
kit receptor kinases. See e.g., Mendel et al., Clin. Cancer Res. 9:
327-37 (2003); O'Farrell et al., Blood 101: 3597-605 (2003); and
Sun et al., J. Med. Chem. 46: 1116-19 (2003). Such compounds
effectively inhibit FLT3 kinase-mediated phosphorylation, cytokine
production, cellular proliferation, resulting in the induction of
apoptosis. See e.g., Spiekermann et al., Blood 101: 1494-1504
(2003). Moreover, such compounds have potent antitumor activity in
vitro and in vivo.
[0234] In some embodiments, the kinase is a class III receptor
tyrosine kinase (RTKIII). In other embodiments, the kinase is a
tyrosine kinase receptor intimately involved in the regulation and
stimulation of cellular proliferation. In still other embodiments,
the kinase is a fms-like tyrosine kinase 3 receptor (FLT-3 kinase).
In this context, inhibition and reduction of the activity of FLT-3
kinase refers to a lower level of measured activity relative to a
control experiment in which the protein, cell, or subject is not
treated with the test compound, whereas an increase in the activity
of FLT-3 kinase refers to a higher level of measured activity
relative to a control experiment. In particular embodiments, the
reduction or increase is at least 10%. One of skill in the art will
appreciate that reduction or increase in the activity of FLT-3
kinase of at least 20%, 50%, 75%, 90% or 100% or any integer
between 10% and 100% may be preferred for particular
applications.
[0235] Compounds provided herein are useful in treating conditions
characterized by inappropriate FLT3 activity such as proliferative
disorders. FLT3 activity includes, but is not limited to, enhanced
FLT3 activity resulting from increased or de novo expression of
FLT3 in cells, increased FLT3 expression or activity, and FLT3
mutations resulting in constitutive activation. The existence of
inappropriate or abnormal FLT3 ligand and FLT3 levels or activity
can be determined using well known methods in the art. For example,
abnormally high FLT3 levels can be determined using commercially
available ELISA kits. FLT3 levels can be determined using flow
cytometric analysis, immunohistochemical analysis, and in situ
hybridization techniques.
[0236] An inappropriate activation of the FLT3 can be determined by
an increase in one or more of the activities occurring subsequent
to FLT3 binding: (1) phosphorylation or autophosphorylation of
FLT3; (2) phosphorylation of a FLT3 substrate, e.g., Stat5, Ras;
(3) activation of a related complex, e.g., PI3K; (4) activation of
an adaptor molecule; and (5) cellular proliferation. These
activities are readily measured by well known methods in the
art.
[0237] Formulations
[0238] The compounds described herein can be used to prepare a
medicament, such as by formulation into pharmaceutical compositions
for administration to a subject using techniques generally known in
the art. A summary of such pharmaceutical compositions may be
found, for example, in Remington's Pharmaceutical Sciences, Mack
Publishing Co., Easton, Pa. The compounds of the invention can be
used singly or as components of mixtures. Preferred forms of the
compounds are those for systemic administration as well as those
for topical or transdermal administration. Formulations designed
for timed release are also within the scope of the invention.
Formulation in unit dosage form is also preferred for the practice
of the invention.
[0239] In unit dosage form, the formulation is divided into unit
doses containing appropriate quantities of one or more compounds.
The unit dosage may be in the form of a package containing discrete
quantities of the formulation. Non-limiting examples are packeted
tablets or capsules, and powders in vials or ampoules.
[0240] The compounds described herein may be labeled isotopically
(e.g. with a radioisotope) or by any other means, including, but
not limited to, the use of chromophores or fluorescent moieties,
bioluminescent labels, or chemiluminescent labels. The compositions
may be in conventional forms, either as liquid solutions or
suspensions, solid forms suitable for solution or suspension in a
liquid prior to use, or as emulsions. Suitable excipients or
carriers are, for example, water, saline, dextrose, glycerol,
alcohols, aloe vera gel, allantoin, glycerin, vitamin A and E oils,
mineral oil, propylene glycol, PPG-2 myristyl propionate, and the
like. Of course, these compositions may also contain minor amounts
of nontoxic, auxiliary substances, such as wetting or emulsifying
agents, pH buffering agents, and so forth.
[0241] Methods for the preparation of compositions comprising the
compounds described herein include formulating the derivatives with
one or more inert, pharmaceutically acceptable carriers to form
either a solid or liquid. Solid compositions include, but are not
limited to, powders, tablets, dispersible granules, capsules,
cachets, and suppositories. Liquid compositions include solutions
in which a compound is dissolved, emulsions comprising a compound,
or a solution containing liposomes, micelles, or nanoparticles
comprising a compound as disclosed herein.
[0242] A carrier of the invention can be one or more substances
which also serve to act as a diluent, flavoring agent, solubilizer,
lubricant, suspending agent, binder, or tablet disintegrating
agent. A carrier can also be an encapsulating material.
[0243] In powder forms of the invention's compositions, the carrier
is preferably a finely divided solid in powder form which is
interdispersed as a mixture with a finely divided powder from of
one or more compound. In tablet forms of the compositions, one or
more compounds is intermixed with a carrier with appropriate
binding properties in suitable proportions followed by compaction
into the shape and size desired. Powder and tablet form
compositions preferably contain between about 5 to about 70% by
weight of one or more compound. Carriers that may be used in the
practice of the invention include, but are not limited to,
magnesium carbonate, magnesium stearate, talc, lactose, sugar,
pectin, dextrin, starch, tragacanth, methyl cellulose, sodium
carboxymethyl cellulose, a low-melting wax, cocoa butter, and the
like.
[0244] The compounds of the invention may also be encapsulated or
microencapsulated by an encapsulating material, which may thus
serve as a carrier, to provide a capsule in which the derivatives,
with or without other carriers, is surrounded by the encapsulating
material. In an analogous manner, cachets comprising one or more
compounds are also provided by the instant invention. Tablet,
powder, capsule, and cachet forms of the invention can be
formulated as single or unit dosage forms suitable for
administration, optionally conducted orally.
[0245] In suppository forms of the compositions, a low-melting wax
such as, but not limited to, a mixture of fatty acid glycerides,
optionally in combination with cocoa butter is first melted. One or
more compounds are then dispersed into the melted material by, as a
non-limiting example, stirring. The non-solid mixture is then
placed into molds as desired and allowed to cool and solidify.
[0246] Non-limiting compositions in liquid form include solutions
suitable for oral or parenteral administration, as well as
suspensions and emulsions suitable for oral administration. Sterile
aqueous based solutions of one or more compounds, optionally in the
presence of an agent to increase solubility of the derivative(s),
are also provided. Non-limiting examples of sterile solutions
include those comprising water, ethanol, and/or propylene glycol in
forms suitable for parenteral administration. A sterile solution of
the invention may be prepared by dissolving one or more compounds
in a desired solvent followed by sterilization, such as by
filtration through a sterilizing membrane filter as a non-limiting
example. In another embodiment, one or more compounds are dissolved
into a previously sterilized solvent under sterile conditions.
[0247] A water based solution suitable for oral administration can
be prepared by dissolving one or more compounds in water and adding
suitable flavoring agents, coloring agents, stabilizers, and
thickening agents as desired. Water based suspensions for oral use
can be made by dispersing one or more compounds in water together
with a viscous material such as, but not limited to, natural or
synthetic gums, resins, methyl cellulose, sodium carboxymethyl
cellulose, polyvinylpyrrolidone, and other suspending agents known
to the pharmaceutical field.
[0248] In therapeutic use, the compounds of the invention are
administered to a subject at dosage levels of from about 0.5 mg/kg
to about 8.0 mg/kg of body weight per day. For example, a human
subject of approximately 70 kg, this is a dosage of from 35 mg to
560 mg per day. Such dosages, however, may be altered depending on
a number of variables, not limited to the activity of the compound
used, the condition to be treated, the mode of administration, the
requirements of the individual subject, the severity of the
condition being treated, and the judgment of the practitioner.
[0249] The foregoing ranges are merely suggestive, as the number of
variables in regard to an individual treatment regime is large, and
considerable excursions from these recommended values are not
uncommon.
[0250] Methods of Use
[0251] By modulating kinase activity, the compounds disclosed
herein can be used to treat a variety of diseases. Suitable
conditions characterized by undesirable protein-kinase activity can
be treated by the compounds presented herein. As used herein, the
term "condition" refers to a disease, disorder, or related symptom
where inappropriate kinase activity is present. In some
embodiments, these conditions are characterized by aggressive
neovasculaturization including tumors, especially acute myelogenous
leukemia (AML), B-precursor cell acute lymphoblastic leukemias,
myelodysplastic leukemias, T-cell acute lymphoblastic leukemias,
and chronic myelogenous leukemias (CMLs). In some embodiments, a
FLT3 modulating compounds may be used to treat tumors. The ability
of compounds that inhibit FLT3 kinase activity to treat tumors has
been established. Compounds having this property include SU5416
(Sugen), PKC412 (Novartis), GTP-14564 and CT53518 (Millennium). See
e.g., Giles et al., Blood 102: 795-801 (2003); Weisberg et al.,
Cancer Cell 1: 433-43 (2002); Murata et al., J. Biol. Chem. 278:
32892-98 (2003); and Kelly et al., Cancer Cell 1: 421-32
(2002).
[0252] Compounds presented herein are useful in the treatment of a
variety of biologically aberrant conditions or disorders related to
tyrosine kinase signal transduction. Such disorders pertain to
abnormal cell proliferation, differentiation, and/or metabolism.
Abnormal cell proliferation may result in a wide array of diseases,
including the development of neoplasia such as carcinoma, sarcoma,
leukemia, glioblastoma, hemangioma, psoriasis, arteriosclerosis,
arthritis and diabetic retinopathy (or other disorders related to
uncontrolled angiogenesis and/or vasculogenesis).
[0253] In various embodiments, compounds presented herein regulate,
modulate, and/or inhibit disorders associated with abnormal cell
proliferation by affecting the enzymatic activity of one or more
tyrosine kinases and interfering with the signal transduced by said
kinase. More particularly, the present invention is directed to
compounds which regulate, modulate said kinase mediated signal
transduction pathways as a therapeutic approach to cure leukemia
and many kinds of solid tumors, including but not limited to
carcinoma, sarcoma, erythroblastoma, glioblastoma, meningioma,
astrocytoma, melanoma and myoblastoma. Indications may include, but
are not limited to brain cancers, bladder cancers, ovarian cancers,
gastric cancers, pancreas cancers, colon cancers, blood cancers,
lung cancers and bone cancers.
[0254] In other embodiments, compounds herein are useful in the
treatment of cell proliferative disorders including cancers, blood
vessel proliferative disorders, fibrotic disorders, and mesangial
cell proliferative disorders. Blood vessel proliferation disorders
refer to angiogenic and vasculogenic disorders generally resulting
in abnormal proliferation of blood vessels. The formation and
spreading of blood vessels, or vasculogenesis and angiogenesis,
respectively, play important roles in a variety of physiological
processes such as embryonic development, corpus luteum formation,
wound healing and organ regeneration. They also play a pivotal role
in cancer development. Other examples of blood vessel proliferation
disorders include arthritis, where new capillary blood vessels
invade the joint and destroy cartilage, and ocular diseases, like
diabetic retinopathy, where new capillaries in the retina invade
the vitreous, bleed and cause blindness. Conversely, disorders
related to the shrinkage, contraction or closing of blood vessels,
such as restenosis, are also implicated.
[0255] Fibrotic disorders refer to the abnormal formation of
extracellular matrix. Examples of fibrotic disorders include
hepatic cirrhosis and mesangial cell proliferative disorders.
Hepatic cirrhosis is characterized by the increase in extracellular
matrix constituents resulting in the formation of a hepatic scar.
Hepatic cirrhosis can cause diseases such as cirrhosis of the
liver. An increased extracellular matrix resulting in a hepatic
scar can also be caused by viral infection such as hepatitis.
Lipocytes appear to play a major role in hepatic cirrhosis. Other
fibrotic disorders implicated include atherosclerosis (see,
below).
[0256] Mesangial cell proliferative disorders refer to disorders
brought about by abnormal proliferation of mesangial cells.
Mesangial proliferative disorders include various human renal
diseases, such as glomerulonephritis, diabetic nephropathy,
malignant nephrosclerosis, thrombotic microangiopathy syndromes,
transplant rejection, and glomerulopathies. The cell proliferative
disorders which are indications of the present invention are not
necessarily independent. For example, fibrotic disorders may be
related to, or overlap, with blood vessel proliferative disorders.
For example, atherosclerosis results, in part, in the abnormal
formation of fibrous tissue within blood vessels.
[0257] Compounds of the invention can be administered to a subject
upon determination of the subject as having a disease or unwanted
condition that would benefit by treatment with said derivative. The
determination can be made by medical or clinical personnel as part
of a diagnosis of a disease or condition in a subject. Non-limiting
examples include determination of a risk of acute myelogenous
leukemia (AML), B-precursor cell acute lymphoblastic leukemias,
myelodysplastic leukemias, T-cell acute lymphoblastic leukemias,
and chronic myelogenous leukemias (CMLs).
[0258] The methods of the invention can comprise the administration
of an effective amount of one or more compounds as disclosed
herein, optionally in combination with one or more other active
agents for the treatment of a disease or unwanted condition as
disclosed herein. The subject is preferably human, and repeated
administration over time is within the scope of the present
invention.
[0259] The present invention thus also provides compounds described
above and their salts or solvates and pharmaceutically acceptable
salts or solvates thereof for use in the prevention or treatment of
disorders mediated by aberrant protein tyrosine kinase activity
such as human malignancies and the other disorders mentioned above.
The compounds of the present invention are especially useful for
the treatment of disorders caused by aberrant kinase activity such
as breast, ovarian, gastric, pancreatic, non-small cell lung,
bladder, head and neck cancers, and psoriasis. The cancers include
hematologic cancers, for example, acute myelogenous leukemia (AML),
B-precursor cell acute lymphoblastic leukemias, myelodysplastic
leukemias, T-cell acute lymphoblastic leukemias, and chronic
myelogenous leukemias (CMLs).
[0260] A further aspect of the invention provides a method of
treatment of a human or animal subject suffering from a disorder
mediated by aberrant protein tyrosine kinase activity, including
susceptible malignancies, which comprises administering to the
subject an effective amount of a compound described above or a
pharmaceutically acceptable salt or solvate thereof.
[0261] A further aspect of the present invention provides the use
of a compound described above, or a pharmaceutically acceptable
salt or solvate thereof, in the preparation of a medicament for the
treatment of cancer and malignant tumors. The cancer can be
stomach, gastric, bone, ovary, colon, lung, brain, larynx,
lymphatic system, genitourinary tract, ovarian, squamous cell
carcinoma, astrocytoma, Kaposi's sarcoma, glioblastoma, lung
cancer, bladder cancer, head and neck cancer, melanoma, ovarian
cancer, prostate cancer, breast cancer, small-cell lung cancer,
leukemia, acute myelogenous leukemia (AML), B-precursor cell acute
lymphoblastic leukemias, myelodysplastic leukemias, T-cell acute
lymphoblastic leukemias, and chronic myelogenous leukemias (CMLs),
glioma, colorectal cancer, genitourinary cancer gastrointestinal
cancer, or pancreatic cancer.
[0262] In accordance with the present invention, compounds provided
herein are useful for preventing and treating conditions associated
with ischemic cell death, such as myocardial infarction, stroke,
glaucoma, and other neurodegenerative conditions. Various
neurodegenerative conditions which may involve apoptotic cell
death, include, but are not limited to, Alzheimer's Disease, ALS
and motor neuron degeneration, Parkinson's disease, peripheral
neuropathies, Down's Syndrome, age related macular degeneration
(ARMD), traumatic brain injury, spinal cord injury, Huntington's
Disease, spinal muscular atrophy, and HIV encephalitis. The
compounds described in detail above can be used in methods and
compositions for imparting neuroprotection and for treating
neurodegenerative diseases.
[0263] The compounds described herein, can be used in a
pharmaceutical composition for the prevention and/or the treatment
of a condition selected from the group consisting of arthritis
(including osteoarthritis, degenerative joint disease,
spondyloarthropathies, gouty arthritis, systemic lupus
erythematosus, juvenile arthritis and rheumatoid arthritis), common
cold, dysmenorrhea, menstrual cramps, inflammatory bowel disease,
Crohn's disease, emphysema, acute respiratory distress syndrome,
asthma, bronchitis, chronic obstructive pulmonary disease,
Alzheimer's disease, organ transplant toxicity, cachexia, allergic
reactions, allergic contact hypersensitivity, cancer (such as solid
tumor cancer including colon cancer, breast cancer, lung cancer and
prostrate cancer; hematopoietic malignancies including leukemias
and lymphomas; Hodgkin's disease; aplastic anemia, skin cancer and
familiar adenomatous polyposis), tissue ulceration, peptic ulcers,
gastritis, regional enteritis, ulcerative colitis, diverticulitis,
recurrent gastrointestinal lesion, gastrointestinal bleeding,
coagulation, anemia, synovitis, gout, ankylosing spondylitis,
restenosis, periodontal disease, epidermolysis bullosa,
osteoporosis, atherosclerosis (including atherosclerotic plaque
rupture), aortic aneurysm (including abdominal aortic aneurysm and
brain aortic aneurysm), periarteritis nodosa, congestive heart
failure, myocardial infarction, stroke, cerebral ischemia, head
trauma, spinal cord injury, neuralgia, neurodegenerative disorders
(acute and chronic), autoimmune disorders, Huntington's disease,
Parkinson's disease, migraine, depression, peripheral neuropathy,
pain (including low back and neck pain, headache and toothache),
gingivitis, cerebral amyloid angiopathy, nootropic or cognition
enhancement, amyotrophic lateral sclerosis, multiple sclerosis,
ocular angiogenesis, corneal injury, macular degeneration,
conjunctivitis, abnormal wound healing, muscle or joint sprains or
strains, tendonitis, skin disorders (such as psoriasis, eczema,
scleroderma and dermatitis), myasthenia gravis, polymyositis,
myositis, bursitis, burns, diabetes (including types I and II
diabetes, diabetic retinopathy, neuropathy and nephropathy), tumor
invasion, tumor growth, tumor metastasis, corneal scarring,
scleritis, immunodeficiency diseases (such as AIDS in humans and
FLV, FIV in cats), sepsis, premature labor, hypoprothrombinemia,
hemophilia, thyroiditis, sarcoidosis, Behcet's syndrome,
hypersensitivity, kidney disease, Rickettsial infections (such as
Lyme disease, Erlichiosis), Protozoan diseases (such as malaria,
giardia, coccidia), reproductive disorders, and septic shock,
arthritis, fever, common cold, pain and cancer in a mammal,
preferably a human, cat, livestock or a dog, comprising an amount
of a compound described herein or a pharmaceutically acceptable
salt thereof effective in such prevention and/or treatment
optionally with a pharmaceutically acceptable carrier.
[0264] A further aspect of the present invention provides the use
of a compound described above, or a pharmaceutically acceptable
salt thereof, in the preparation of a medicament for the treatment
of psoriasis.
[0265] As one of skill in the art will recognize, the compounds can
be administered before, during or after the occurrence of a
condition or a disease, and the timing of administering the
composition containing a compound can vary. Thus, for example, the
compounds can be used as a prophylactic and can be administered
continuously to subjects with a propensity to conditions and
diseases in order to prevent the occurrence of the disorder. The
compounds and compositions can be administered to a subject during
or as soon as possible after the onset of the symptoms. The
administration of the compounds can be initiated within the first
48 hours of the onset of the symptoms, preferably within the first
48 hours of the onset of the symptoms, more preferably within the
first 6 hours of the onset of the symptoms, and most preferably
within 3 hours of the onset of the symptoms. The initial
administration can be via any route practical, such as, for
example, an intravenous injection, a bolus injection, infusion over
5 min. to about 5 hours, a pill, a capsule, transdermal patch,
buccal delivery, and the like, or a combination thereof. A compound
is preferably administered as soon as is practicable after the
onset of a condition or a disease is detected or suspected, and for
a length of time necessary for the treatment of the disease, such
as, for example, from about 1 month to about 3 months. As one of
skill in the art will recognize, the length of treatment can vary
for each subject, and the length can be determined using the known
criteria. For example, the compound or a formulation containing the
compound can be administered for at least 2 weeks, preferably about
1 month to about 5 years, and more preferably from about 1 month to
about 3 years.
[0266] Kits/Articles of Manufacture
[0267] For use in the therapeutic applications described herein,
kits and articles of manufacture are also within the scope of the
invention. Such kits can comprise a carrier, package, or container
that is compartmentalized to receive one or more containers such as
vials, tubes, and the like, each of the container(s) comprising one
of the separate elements to be used in a method of the invention.
Suitable containers include, for example, bottles, vials, syringes,
and test tubes. The containers can be formed from a variety of
materials such as glass or plastic.
[0268] For example, the container(s) can comprise one or more
compounds of the invention, optionally in a composition or in
combination with another agent as disclosed herein. The
container(s) optionally have a sterile access port (for example the
container can be an intravenous solution bag or a vial having a
stopper pierceable by a hypodermic injection needle). Such kits
optionally comprising a compound with an identifying description or
label or instructions relating to its use in the methods of the
present invention.
[0269] A kit of the invention will typically may comprise one or
more additional containers, each with one or more of various
materials (such as reagents, optionally in concentrated form,
and/or devices) desirable from a commercial and user standpoint for
use of a compound of the invention. Non-limiting examples of such
materials include, but not, limited to, buffers, diluents, filters,
needles, syringes; carrier, package, container, vial and/or tube
labels listing contents and/or instructions for use, and package
inserts with instructions for use. A set of instructions will also
typically be included.
[0270] A label can be on or associated with the container. A label
can be on a container when letters, numbers or other characters
forming the label are attached, molded or etched into the container
itself; a label can be associated with a container when it is
present within a receptacle or carrier that also holds the
container, e.g., as a package insert. A label can be used to
indicate that the contents are to be used for a specific
therapeutic application. The label can also indicate directions for
use of the contents, such as in the methods described herein.
[0271] The terms "kit" and "article of manufacture" may be used as
synonyms.
EXAMPLES
[0272] The present invention is further illustrated by the
following examples, which should not be construed as limiting in
any way. The experimental procedures to generate the data shown are
discussed in more detail below. For all formulations herein,
multiple doses may be proportionally compounded as is known in the
art. The coatings, layers and encapsulations are applied in
conventional ways using equipment customary for these purposes.
[0273] The invention has been described in an illustrative manner,
and it is to be understood that the terminology used is intended to
be in the nature of description rather than of limitation. Thus, it
will be appreciated by those of skill in the art that conditions
such as choice of solvent, temperature of reaction, volumes,
reaction time may vary while still producing the desired compounds.
In addition, one of skill in the art will also appreciate that many
of the reagents provided in the following examples may be
substituted with other suitable reagents. See, e.g., Smith &
March, Advanced Organic Chemistry, 5.sup.th ed. (2001).
Example A
Synthesis of Isoxazole-Ureas
[0274] 47
[0275] A mixture of amine 1 (1 eq) in dry THF is stirred at room
temperature under argon for an hour. Then the stirred suspension is
cooled to 0.degree. C. and to it is added dropwise a solution of
phosgene or disuccinimidyl carbonate or carbonyl diimidazole (1.2
eq). The reaction is stirred at 0.degree. C. for half an hour. An
isoxazol-amine 2 in THF is added dropwise and the reaction is
allowed to warm to room temperature and stirred overnight. The
solvent is removed and extracted with ethyl acetate and water. The
organic layer is dried over magnesium sulfate and solvent removed,
and the product 3 purified by HPLC. 48
[0276] Alternatively, to a stirring solution of an isoxazol-amine 2
(1 eq) in THF, a mixture of 4-nitrophenyl chloroformate (1.2 eq)
and triethyl amine (1.2 eq) is added dropwise at 0.degree. C. The
reaction is stirred for two hours at room temperature and the
aniline 1 is added. The reaction is refluxed to 80.degree. C. for
six hours. The mixture is cooled to room temperature and poured
into water and extracted with ethyl acetate and dried over
magnesium sulfate, and the product 3 purified by HPLC. 49
[0277] Alternatively, amine 1 (1 eq) is dissolved in toluene at
room temperature and stirred for 10 minutes. Then
3-tert-butyl-isoxazol-5-yl isocyanate 4 in toluene is added and
heated at 80.degree. C. for 4 hours. The solvent is removed and the
crude mixture is purified by HPLC to obtain 5. 50
Synthesis of Compound A1:
1-(5-tert-butylisoxazol-3-yl)-3-(4-chloro-3-(tri-
fluoromethyl)phenyl)urea
[0278] 51
[0279] To a stirring solution of 5-tert-Butyl-isoxazol-3-ylamine
(250 mg, 1 eq) in dry toluene at 0.degree. C. trichloromethyl
chloroformate (1.1 eq) was added dropwise. The reaction stirred at
0.degree. C. and allowed to warm to room temperature overnight. The
solvent was removed and the mixture was recrystallized in ethyl
acetate. The solid was filtered off and washed with cold ethyl
acetate. Yield: 242 mg (83%). 52
[0280] To a flask 5-tert-Butyl-3-isocyanato-isoxazole (242 mg, 1
eq) and substituted aniline (159 mg, 1 eq) was added and dissolved
in toluene. The reaction was allowed to stir at 50.degree. C. for
three hours. The solvent was removed and the mixture was purified
by HPLC. Yield: 188 mg (47%).
[0281] Compounds A2 through A57 were synthesized in a manner
analogous to Compound A1 using similar starting materials and
reagents. The structures are shown below in Table A:
2TABLE A NO. CHEMICAL STRUCTURE A1 53 A2 54 A3 55 A4 56 A5 57 A6 58
A7 59 A8 60 A9 61 A10 62 A11 63 A12 64 A13 65 A14 66 A15 67 A16 68
A17 69 A18 70 A19 71 A20 72 A21 73 A22 74 A23 75 A24 76 A25 77 A26
78 A27 79 A28 80 A29 81 A30 82 A31 83 A32 84 A33 85 A34 86 A35 87
A36 88 A37 89 A38 90 A39 91 A40 92 A41 93 A42 94 A43 95 A44 96 A45
97 A46 98 A47 99 A48 100 A49 101 A50 102 A51 103 A52 104 A53 105
A54 106 A55 107 A56 108 A57 109
Example B
Synthesis of Alkyl-Ureas
Synthesis of Compound B1:
1-(4-methoxybenzyl)-3-(5-tert-butylisoxazol-3-yl- )urea
[0282] 110
[0283] To a flask 5-tert-Butyl-3-isocyanato-isoxazole (242 mg, 1
eq) and substituted benzylamine (1 eq) was added and dissolved in
toluene. The reaction was allowed to stir at 50.degree. C. for
three hours. The solvent removed and the mixture was purified by
HPLC. Yield: 188 mg (47%).
[0284] Compounds B2 through B8 were synthesized in a manner
analogous to Compound B1 using similar starting materials and
reagents. The structures are shown below in Table B:
3TABLE B NO. CHEMICAL STRUCTURE B1 111 B2 112 B3 113 B4 114 B5 115
B6 116 B7 117 B8 118
Example C
Synthesis of Reactive Ureas
Synthesis of Compound C1:
1-(5-tert-butylisoxazol-3-yl)-3-(4-aminophenyl)u- rea
[0285] 119
[0286] To a flask 5-tert-Butyl-3-isocyanato-isoxazole (242 mg, 1
eq) and substituted aniline (159 mg, 1 eq) was added and dissolved
in toluene. The reaction was allowed to stir at 50.degree. C. for
three hours. The solvent removed and the mixture was purified by
HPLC. Yield: 188 mg (47%).
[0287] Compounds C2 through C3 were synthesized in a manner
analogous to Compound C1 using similar starting materials and
reagents. The structures are shown below in Table C:
4TABLE C NO. CHEMICAL STRUCTURE C1 120 C2 121 C3 122
Example D
Synthesis of Substituted-Pyrazole Ureas
Synthesis of Compound D1:
1-(3-tert-butyl-1-p-tolyl-1H-pyrazol-5-yl)-3-(4--
isopropylphenyl)urea
[0288] 123
[0289] To a stirring solution of
5-tert-Butyl-2-p-tolyl-2H-pyrazol-3-ylami- ne (250 mg, 1 eq) in dry
toluene at 0.degree. C. trichloromethyl chloroformate (1.1 eq) was
added dropwise. The reaction stirred at 0.degree. C. and allowed to
warm to room temperature overnight. The solvent was removed and the
mixture was recrystallized in ethyl acetate. The solid was filtered
off and washed with cold ethyl acetate. Yield: 242 mg (83%).
124
[0290] To a flask 3-tert-Butyl-5-isocyanato-1-p-tolyl-1H-pyrazole
(242 mg, 1 eq) and substituted aniline (159 mg, 1 eq) was added and
dissolved in toluene. The reaction was allowed to stir at
50.degree. C. for three hours. The solvent was removed and the
mixture was purified by HPLC. Yield: 188 mg (47%).
[0291] Compounds D2 through D22 were synthesized in a manner
analogous to Compound D1 using similar starting materials and
reagents. The structures are shown below in Table D:
5TABLE D NO. CHEMICAL STRUCTURE D1 125 D2 126 D3 127 D4 128 D5 129
D6 130 D7 131 D8 132 D9 133 D10 134 D11 135 D12 136 D13 137 D14 138
D15 139 D16 140 D17 141 D18 142 D19 143 D20 144 D21 145 D22 146
Example E
Exemplary Synthesis of Cyclic Ureas
[0292] 147
[0293] To the urea 6 is added NaH (2.5 eq) in DMF and the reaction
is stirred at 40.degree. C. for 1 hour. Then 1 eq of
1,3-dibromopropane is added and the reaction heated to 80.degree.
C. for 8 hours, then cooled, the solvent removed in vacuo and the
product 7 purified by HPLC.
Synthesis of Compound E1:
3-(3-tert-butyl-1-p-tolyl-1H-pyrazol-5-yl)-1-(4--
(benzyloxy)phenyl)-tetrahydropyrimidin-2(1H)-one
[0294] 148149
[0295] To a stirring solution of 4-aminophenol (1 g, 1 eq) in 20 mL
THF at 0.degree. C. di-tert-butyl dicarbonate (2 g, 1 eq) in 3 mL
THF was slowly added dropwise over 30 minutes. The reaction stirred
at 0.degree. C. and allowed to warm to room temperature overnight.
The solvent removed and diluted with ethyl acetate. It was then
extracted with water three times, and the organic layer was dried
over magnesium sulfate. It was recrystallized in dichloromethane.
Yield: 1.5 g (79%).
[0296] Bocaminophenol (0.5 g, 1 eq), benzyl bromide (0.45 g, 1 eq),
and cesium carbonate (1.94 g, 2.5 eq) was dissolved in 30 mL
dimethylformamide. The reaction was allowed to stir at 45.degree.
C. overnight. The solvent was removed and dissolved in ethyl
acetate and water. It was extracted with ethyl acetate three times.
The organic layer was washed with 1N sodium hydroxide and dried
with magnesium sulfate and solvent was removed. It was purified by
column chromatography. Yield: 0.44 g (58%).
[0297] [4-(benzyloxy)-phenyl]-carbamic acid tert-butyl ester (0.44
g) was dissolved in 6 mL dichloromethane and 2 mL trifluoroacetic
acid was added. The reaction stirred at room temperature for 1
hour. The excess trifluoroacetic acid was removed in vivo. Yield:
0.12 g (42%).
[0298] 4-(benzyloxy)-phenylamine (0.12 g, 1 eq) was mixed with
5-tert-Butyl-2-p-tolyl-2H-pyrazole-3-carbonitrile N-oxide (0.9 g 1
eq) and dissolved in dry toluene. The reaction stirred at
80.degree. C. overnight. The solvent was removed and purified by
HPLC. Yield: 85 mg (31%) 150
[0299] To 6 is added NaH (2.5 eq) in DMF and the reaction is
stirred at 40.degree. C. for 1 hour. Then 1 eq of
1,3-dibromopropane is added and the reaction heated to 80.degree.
C. for 8 hours, then cooled, the solvent removed in vacuo and the
product 7 purified by HPLC.
[0300] Compounds E2 through E5 were synthesized in a manner
analogous to Compound E1 using similar starting materials and
reagents. The structures are shown below in Table E:
6TABLE E NO. CHEMICAL STRUCTURE E1 151 E2 152 E3 153 E4 154 E5
155
Example F
Conversion of Ureas to Thioureas
[0301] 156
[0302] Lawesson's reagent is added to starting urea in toluene and
the reaction heated to 100.degree. C. for 8 hours, then cooled, the
solvent removed in vacuo and the thiourea purified by HPLC.
Synthesis of Compound F1:
1-(5-tert-butylisoxazol-3-yl)-3-(4-morpholinophe- nyl)thiourea
[0303] 157
[0304] To a flask 5-tert-Butyl-3-isocyanato-isoxazole (242 mg, 1
eq) and substituted aniline (159 mg, 1 eq) was added and dissolved
in toluene. The reaction was allowed to stir at 50.degree. C. for
three hours. The solvent removed and the mixture was purified by
HPLC. Yield: 188 mg (47%) 158
[0305] Lawesson's reagent is added to starting urea in toluene and
the reaction heated to 100.degree. C. for 8 hours, then cooled, the
solvent removed in vacuo and the thiourea purified by HPLC.
[0306] Compounds F2 through F9 were synthesized in a manner
analogous to Compound F1 using similar starting materials and
reagents. The structures are shown below in Table F:
7TABLE F NO. CHEMICAL STRUCTURE F1 159 F2 160 F3 161 F4 162 F5 163
F6 164 F7 165 F8 166 F9 167
Example G
Exemplary Synthesis of Ureas with Ether Linkers
[0307] 168
[0308] To a stirring solution of the amine 4 (1 eq) in toluene at
room temperature is added the isocyanate 8 and heated at 80.degree.
C. overnight. The solvent is removed, and the product 9 is purified
by HPLC. Compound C, compound A, compound B, compound D and other
compounds in the benzoloxy series were made by the general method
described above. 169
[0309] To a stirring solution of 4-aminophenol (1 eq) in THF at
0.degree. C. di-tert-butyl dicarbonate (1 eq) in THF was slowly
added dropwise over 30 minutes. The reaction stirred at 0.degree.
C. and allowed to warm to room temperature overnight. The solvent
removed and diluted with ethyl acetate. It was then extracted with
water three times, and the organic layer was dried over magnesium
sulfate. It was recrystallized in dichloromethane.
[0310] To a stirring solution of sodium hydride (1.2 eq) in DMF,
the Boc-aminophenol (1 eq) was added dropwise at 0.degree. C. and
stirred to room temperature for one hour. Then the substituted
benzyl halide (1 eq) in THF was added dropwise at 0.degree. C. The
reaction was allowed to stir at 40.degree. C. overnight. The
solvent was removed and dissolved in ethyl acetate and water. It
was extracted with ethyl acetate three times. The organic layer was
washed with 1N sodium hydroxide and dried with magnesium sulfate
and solvent was removed. It was purified by column
chromatography.
[0311] The protected substituted benzyloxyaniline was dissolved in
dichloromethane and trifluoroacetic acid was added. The reaction
stirred at room temperature for 1 hour. The excess trifluoroacetic
acid was removed in vivo.
[0312] The substituted benzyloxyaniline (1 eq) was mixed with
5-tert-Butyl-3-isocyanato-isoxazole (1 eq) and dissolved in dry
toluene. The reaction stirred at 80.degree. C. overnight. The
solvent was removed and purified by HPLC.
Synthesis of Compound G1:
1-(4-(3-(pyridin-4-yl)propoxy)phenyl)-3-(5-tert--
butylisoxazol-3-yl)urea
[0313] 170
[0314] To a stirring solution of 5-tert-Butyl-isoxazol-3-ylamine
(250 mg, 1 eq) in dry toluene at 0.degree. C. trichloromethyl
chloroformate (1.1 eq) was added dropwise. The reaction stirred at
0.degree. C. and allowed to warm to room temperature overnight. The
solvent was removed and the mixture was recrystallized in ethyl
acetate. The solid was filtered off and washed with cold ethyl
acetate. Yield: 242 mg (83%).
[0315] To a flask 5-tert-Butyl-3-isocyanato-isoxazole (242 mg, 1
eq) and 4-aminophenol (159 mg, 1 eq) was added and dissolved in
toluene. The reaction was allowed to stir at 50.degree. C. for
three hours. The solvent removed and the mixture was purified by
HPLC. Yield: 188 mg (47%), LC/MS [MH.sup.+] 276.
[0316] In a dry flask flushed with nitrogen gas
1-(5-tert-Butyl-isoxazol-3- -yl)-3-(4-hydroxy-phenyl)-urea (100 mg,
1 eq), 3-Pyridin-4-yl-propan-1-ol (200 mg, 1 eq) and
triphenylphosphine (143 mg, 1.5 eq) was added and then dissolved
with THF. The flask was cooled to 0.degree. C. and diethyl
azodicarboxylate (95 mg, 1.5 eq) was added dropwise. The reaction
stirred overnight at room temperature. The THF was removed and the
mixture was purified by HPLC. Yield: 26 mg (18%), LC/MS [MH.sup.+]
395.
Synthesis of Compound G12:
1-(4-(2-fluorobenzyloxy)phenyl)-3-(5-tert-butyl-
isoxazol-3-yl)urea
[0317] 171
[0318] To a stirring solution of 4-aminophenol (1 g. 1 eq) in 20 mL
THF at 0.degree. C. di-tert-butyl dicarbonate (2 g, 1 eq) in 3 mL
THF was slowly added dropwise over 30 minutes. The reaction stirred
at 0.degree. C. and allowed to warm to room temperature overnight.
The solvent removed and diluted with ethyl acetate. It was then
extracted with water three times, and the organic layer was dried
over magnesium sulfate. It was recrystallized in dichloromethane.
Yield: 1.5 g (79%).
[0319] Bocaminophenol (0.5 g, 1 eq), 2-fluorobenzyl bromide (0.45
g, 1 eq), and cesium carbonate (1.94 g, 2.5 eq) was dissolved in 30
mL dimethylformamide. The reaction was allowed to stir at
45.degree. C. overnight. The solvent was removed and dissolved in
ethyl acetate and water. It was extracted with ethyl acetate three
times. The organic layer was washed with 1N sodium hydroxide and
dried with magnesium sulfate and solvent was removed. It was
purified by column chromatography. Yield: 0.44 g (58%).
[0320] [4-(2-Fluoro-benzyloxy)-phenyl]-carbamic acid tert-butyl
ester (0.44 g) was dissolved in 6 mL dichloromethane and 2 mL
trifluoroacetic acid was added. The reaction stirred at room
temperature for 1 hour. The excess trifluoroacetic acid was removed
in vivo. Yield: 0.12 g (42%), LC/MS [MH.sup.+] 218.
[0321] 4-(2-Fluoro-benzyloxy)-phenylamine (0.12 g, 1 eq) was mixed
with 5-tert-Butyl-3-isocyanato-isoxazole (0.9 g 1 eq) and dissolved
in dry toluene. The reaction stirred at 80.degree. C. overnight.
The solvent was removed and purified by HPLC. Yield: 85 mg (31%),
LC/MS [MH.sup.+] 384.
[0322] Compounds G2 through G57 were synthesized in a manner
analogous to Compound G1 and G12 using similar starting materials
and reagents. The structures are shown below in Table G:
8TABLE G NO. CHEMICAL STRUCTURE G1 172 G2 173 G3 174 G4 175 G5 176
G6 177 G7 178 G8 179 G9 180 G10 181 G11 182 G12 183 G13 184 G14 185
G15 186 G16 187 G17 188 G18 189 G19 190 G20 191 G21 192 G22 193 G23
194 G24 195 G25 196 G26 197 G27 198 G28 199 G29 200 G30 201 G31 202
G32 203 G33 204 G34 205 G35 206 G36 207 G37 208 G38 209 G39 210 G40
211 G42 212 G44 213 G46 214 G48 215 G49 216 G50 217 G51 218 G52 219
G53 220 G54 221 G55 222 G56 223 G57 224
Example H
Exemplary Synthesis of Ureas with Ether Linkers
Synthesis of Compound H1:
1-(4-(benzyloxy)phenyl)-3-(1-phenyl-1H-pyrazol-5- -yl)urea
[0323] 225
[0324] Compounds H1 through H10 were synthesized in a manner
analogous to compound G1. Compounds H11 through H17 were
synthesized in a manner analogous to Compound A1 using similar
starting materials and reagents. The structures are shown below in
Table H:
9TABLE H NO. CHEMICAL STRUCTURE H1 226 H2 227 H3 228 H4 229 H5 230
H6 231 H7 232 H8 233 H9 234 H10 235 H11 236 H12 237 H13 238 H14 239
H15 240 H16 241 H17 242
Example I
Exemplary Synthesis of Ureas with Ether Linkers
Synthesis of Compound I1:
1-(4-(benzyloxy)phenyl)-3-(4-methylthiazol-2-yl)- urea
[0325] 243
[0326] Compounds I1 through I4 were synthesized in a manner
analogous to Compound G1 using similar starting materials and
reagents. The structures are shown below in Table I:
10TABLE I NO. CHEMICAL STRUCTURE I1 244 I2 245 I3 246 I4 247
Example J
Synthesis of N-Substituted Ureas
[0327] 248
[0328] A solution of an isoxazole-3-isocyanate in dimethylacetamide
is added to a solution of 4-alkoxy-N-alkylbenzenamine in
dimethylacetamide, and the mixture is heated at 80.degree. C.
overnight. After cooling to room temperature, 20 ml water is added
and the mixture is extracted with 3.times.30 ml EtOAc. The combined
organic phases are washed with brine, dried over magnesium sulfate,
and evaporated. Purification of the product is accomplished by
flash chromatography (silica gel, hexanes, 0-50% EtOAc).
Synthesis of Compound J1:
1-(5-tert-butylisoxazol-3-yl)-3-(4-methoxyphenyl-
)-3-methylurea
[0329] 249
[0330] A solution of 5-tert-butyl-isoxazole-3-isocyanate (166 mg, 1
mmol) in 0.5 ml dimethylacetamide was added to a solution of
4-methoxy-N-methylaniline in 0.5 ml dimethylacetamide (137 mg, 1
mmol), and the mixture was heated at 80.degree. C. overnight. After
cooling to room temperature, 20 ml water was added and the mixture
was extracted with 3.times.30 ml EtOAc. The combined organic phases
were washed with brine, dried over magnesium sulfate, and
evaporated. Purification of the product,
N-(methyl)(4-methoxyphenyl)-N-(5-t-butyl-3-isoxazolyl) urea, was
accomplished by flash chromatography (silica gel, hexanes, 0-50%
EtOAc).
[0331] Compounds J2 through J15 were synthesized in a manner
analogous to Compound J1 using similar starting materials and
reagents. The structures are shown below in Table J:
11TABLE J NO. CHEMICAL STRUCTURE J1 250 J2 251 J3 252 J4 253 J5 254
J6 255 J7 256 J8 257 J9 258 J10 259 J11 260 J12 261 J13 262 J14 263
J15 264
Example K
Synthesis of Compounds with Ether-Alkyl Chains
[0332] 265
[0333] The following general procedure was used to synthesize
compounds having the ethylene, propylene, or butylenes linkers. To
a stirring solution of 4-aminophenol (1 eq) in THF at 0.degree. C.,
di-tert-butyl dicarbonate (1 eq) in THF was slowly added dropwise
over 30 minutes. The reaction was stirred at 0.degree. C. and
allowed to warm to room temperature overnight. The solvent removed
and diluted with ethyl acetate. It was then extracted with water
three times, and the organic layer was dried over magnesium
sulfate. The solid was recrystallized from dichloromethane.
[0334] In a dry flask flushed with nitrogen gas bocaminophenol (1
eq), substituted alcohol (1 eq) and triphenylphosphine (1.5 eq) was
added and then dissolved with THF. The flask was cooled to
0.degree. C. and diethyl azodicarboxylate (1.5 eq) was added
dropwise. The reaction stirred overnight at room temperature. THF
was removed under vacuum and the mixture was purified by HPLC.
[0335] The protected substituted ethyloxyaniline was dissolved in
dichloromethane and trifluoroacetic acid was added. The reaction
stirred at room temperature for 1 hour. The excess trifluoroacetic
acid was removed in vivo.
[0336] The substituted ethyloxyaniline (1 eq) was mixed with
5-tert-Butyl-3-isocyanato-isoxazole (1 eq) and dissolved in dry
toluene. The reaction stirred at 50.degree. C. overnight. The
solvent was removed and purified by HPLC. Compound W, compound X,
and others in the series were synthesized using the procedure
described above.
Synthesis of Compound K1:
1-(4-(2-morpholinoethoxy)naphthyl)-3-(3-tert-but-
ylisoxazol-5-yl)urea
[0337] 266
[0338] To a mixture of 4-amino-1-naphthol hydrochloride (15 g) in
100 mL dry THF at -78 C was added dropwise over 1 h n-BuLi (43 mL
of a 1.6M solution in hexanes). After the addition was complete the
mixture was allowed to warm to room temperature and then cooled to
-78 C aand di-tert-butyl dicarbonate (Boc.sub.2O) (16.5 g in 100 mL
THF was added over a period of 20 min). The mixture was slowly
warmed to room temperature and stirred for 24 h and then most of
the volatiles removed in vacuo. The residue was diluted with ethyl
acetate and washed with water (3.times.100 mL) and brine (100 mL)
and filtered through celite and dried over magnesium sulphate.
Column chromatography (30% EtOAc/hexanes) gave 20 g of pure
product. 267
[0339] To a solution of (4-Hydroxy-naphthalen-1-yl)-carbamic acid
tert-butyl ester (3 g) and 4-(2-Chloro-ethyl)-morpholine
hydrochloride (2.22 g) in 25 mL of acetonitrile was added powdered
potassium carbonate (6 g) and the solution heated overnight at 80
C. It was cooled, diluted with EtOAc and water. The organic layer
was washed with water, brine and dried over anhydrous magnesium
sulfate and the volatiles removed in vacuo. Purification by silica
gel chromatography (10% EtOAc/hexanes) gave 2.3 g of pure product.
268
[0340] To a flask 5-tert-Butyl-3-isocyanato-isoxazole (242 mg, 1
eq) and substituted aniline (159 mg, 1 eq) was added and dissolved
in toluene. The reaction was allowed to stir at 50.degree. C. for
three hours. The solvent removed and the mixture was purified by
HPLC. Yield: 188 mg (47%).
[0341] Compound K2 was synthesized in a manner analogous to
Compound K1 using similar starting materials and reagents. The
structures are shown below in Table K:
12TABLE K NO. CHEMICAL STRUCTURE K1 269 K2 270
Example L
Exemplary Synthesis of Isoxazole-Urea
Synthesis of Compound L1:
1,3-bis(5-tert-butylisoxazol-3-yl)urea
[0342] 271
[0343] A mixture of amine 5-tert-butyl-isoxazol-3-ylamine (1 eq) in
dry THF is stirred at room temperature under argon for an hour.
Then the stirred suspension is cooled to 0.degree. C. and to it is
added dropwise a solution of phosgene or disuccinimidyl carbonate
or carbonyl diimidazole (1.2 eq). The reaction is stirred at
0.degree. C. for half an hour. Then 5-tert-butyl-isoxazol-3-ylamine
in THF is added dropwise and the reaction is allowed to warm to
room temperature and stirred overnight. The solvent is removed and
extracted with ethyl acetate and water. The organic layer is dried
over magnesium sulfate and solvent removed, and the
1,3-bis(5-tert-butylisoxazol-3-yl)urea product purified by
HPLC.
Example M
Exemplary Synthesis of Pyrimidine Containing Compounds
[0344] 272
[0345] Dichloro-dimethoxyquinazoline was heated for 3 hours at
80.degree. C. with one equivalent p-nitroaniline in n-butanol.
After cooling to room temperature, isopropanol was added and the
insoluble product was collected by filtration. Reduction was
accomplished using 10% Pd/C in methanol at 50 psi in the presence
of hydrochloric acid. The resulting aniline was reacted with the
oxazole isocyanate in DMA at 80.degree. C. in the presence of DIEA.
The urea product was purified by HPLC. 273
[0346] A mixture of aminourea hydrochloride (155 mg, 0.5 mmol),
halopurine/haloquinazoline/halopyrimidine (0.5 mmol), and DIEA (90
.mu.l, 0.5 mmol) in n-butanol (2-5 ml) was heated at 100.degree. C.
for 1-16 h. The product was purified by HPLC. 274
[0347] A mixture of aminourea (274 mg, 1 mmol), paraformaldehyde
(30 mg, 0.33 mmol), sodium triacetoxyborohydride (636 mg, 3 mmol)
and 10 drops glacial acetic acid was stirred at room temperature
over night. The solvent was evaporated completely and the residue
was treated with sat. aq. NaHCO.sub.3. The solids were collected by
filtration and purified via HPLC. The resulting methylaniline was
used in amide formations and aryl aminations as descibed above.
Example N
Exemplary Synthesis of Compounds Containing Amide Linkers
[0348] 275
[0349] 1 gm. (6 mmol) of 5-tert-butyl-isoxazole-3-isocyanate and
0.83 gm (6 mmol) 4-nitro-phenylamine were dissolved in 20 ml dry
toluene and stirred at 80.degree. C. for 24 h. The resulting
suspension was cooled to room temperature and filtered off to give
the title compound as a yellow solid. The product was used in the
next step without further purification. Yield: 1.7 g (92%), LC/MS
[MH.sup.+] 305. 276
[0350] 1.5 gm of
1-(5-tert-butyl-isoxazol-3-yl)-3-(4-nitro-phenyl)-urea was
dissolved in 50 ml THF and 0.1 g of 10% Pd/C was added. The
solution was stirred under hydrogen at 50 psi. for 24 h than
filtered trough Celite pad. The organic solvent was evaporated
under vacuum and the resulting residue was triturated with ethyl
acetate. Yield: 1.3 g (96%), LC/MS [MH.sup.+] 275. 277
[0351] 1 equivalent of the carboxylic acid and 1.1 equivalent of
CDI were dissolved in dry DMF and stirred at 40.degree. C. for 2 h,
than 1 equivalent of aniline was added. The reaction mixture was
stirred at 40.degree. C. overnight and the final product was
purified by preparative HPLC. 278
[0352] Alternatively, 1 equivalent of the carboxylic acid and 1.1
equivalent of thionyl chloride were heated in a sealed tube at
50.degree. C. for 3 h. The excess thionyl chloride was evaporated,
1 equivalent of aniline in DMF was added, and the solution stirred
at room temperature for 8 h. The final product was purified by
preparative HPLC.
[0353] Compounds N1 through N189 were synthesized in a manner
analogous to one of the above procedures using similar starting
materials and reagents. The structures are shown below in Table
N:
13TABLE N NO. CHEMICAL STRUCTURE N1 279 N2 280 N3 281 N4 282 N5 283
N6 284 N7 285 N8 286 N9 287 N10 288 N11 289 N12 290 N13 291 N14 292
N15 293 N16 294 N17 295 N18 296 N19 297 N20 298 N21 299 N22 300 N23
301 N24 302 N25 303 N26 304 N27 305 N28 306 N29 307 N30 308 N31 309
N32 310 N33 311 N34 312 N35 313 N36 314 N37 315 N38 316 N39 317 N40
318 N41 319 N42 320 N43 321 N44 322 N45 323 N47 324 N48 325 N49 326
N50 327 N51 328 N52 329 N53 330 N54 331 N55 332 N56 333 N57 334 N58
335 N59 336 N60 337 N61 338 N62 339 N63 340 N64 341 N65 342 N66 343
N67 344 N68 345 N69 346 N70 347 N71 348 N72 349 N73 350 N74 351 N75
352 N76 353 N77 354 N78 355 N79 356 N80 357 N81 358 N82 359 N83 360
N84 361 N85 362 N86 363 N87 364 N88 365 N89 366 N90 367 N91 368 N92
369 N93 370 N94 371 N95 372 N96 373 N97 374 N98 375 N99 376 N100
377 N101 378 N102 379 N103 380 N104 381 N105 382 N106 383 N107 384
N108 385 N109 386 N110 387 N111 388 N112 389 N113 390 N114 391 N115
392 N116 393 N117 394 N118 395 N119 396 N120 397 N121 398 N122 399
N123 400 N124 401 N125 402 N126 403 N127 404 N128 405 N129 406 N130
407 N131 408 N132 409 N133 410 N134 411 N135 412 N136 413 N137 414
N138 415 N139 416 N140 417 N141 418 N142 419 N143 420 N144 421 N145
422 N146 423 N147 424 N148 425 N149 426 N150 427 N151 428 N152 429
N153 430 N154 431 N155 432 N156 433 N157 434 N158 435 N159 436 N160
437 N161 438 N162 439 N163 440 N164 441 N165 442 N166 443 N167 444
N168 445 N169 446 N170 447 N171 448 N172 449 N173 450 N174 451 N175
452 N176 453 N177 454 N178 455 N179 456 N180 457 N181 458 N182 459
N183 460 N184 461 N185 462 N186 463 N187 464 N188 465 N189 466
Example O
Synthesis of Compounds with Amine Linkers
[0354] 467
[0355] Dichloro-dimethoxyquinazoline was heated 3 h at 80.degree.
C. with one equivalent p-nitroaniline in n-butanol. After cooling
to room temperature, isopropanol was added and the insoluble
product was collected by filtration. Reduction was accomplished
using 10% Pd/C in methanol at 50 psi in the presence of
hydrochloric acid. The resulting aniline was reacted with the
oxazole isocyanate in DMA at 80.degree. C. in the presence of DIEA.
The urea product was purified by HPLC.
Synthesis of Compound O1:
1-(4-(2-chloro-6,7-dimethoxyquinazolin-4-ylamino-
)phenyl)-3-(5-tert-butylisoxazol-3-yl)urea
[0356] 468
[0357] Dichloro-dimethoxyquinazoline was heated 3 h at 80.degree.
C. with one equivalent p-nitroaniline in n-butanol. After cooling
to room temperature, isopropanol was added and the insoluble
product was collected by filtration. Reduction was accomplished
using 10% Pd/C in methanol at 50 psi in the presence of
hydrochloric acid. The resulting aniline was reacted with the
oxazole isocyanate in DMA at 80.degree. C. in the presence of DIEA.
The urea product was purified by HPLC.
[0358] Compounds O2 through O10 were synthesized in a manner
analogous to Compound O1 using similar starting materials and
reagents. The structures are shown below in Table O:
14TABLE O NO. CHEMICAL STRUCTURE O1 469 O2 470 O3 471 O4 472 O5 473
O6 474 O7 475 O8 476 O9 477 O10 478
Example P
Synthesis of Chalcones
[0359] 479
[0360] Acetylphenylurea (obtained from either reacting
p-aminoacetophenone with oxazole isocyanate in toluene or reacting
p-acetylphenyl isocyanate with oxazole amine in toluene) was
reacted with e.g. 4-pyridine carboxaldehyde analogous to a
literature procedure (Zhang et al., Chem. Lett. 2003, 32,
966-967).
[0361] Chalcone intermediates were further modified according to
procedures described in the literature. See Powers et al.,
Tetrahedron, 1998, 54, 4085-4096; Katritzky et al. Org. Lett. 2000,
2, 429-431.
Compound P1:
1-(5-isopropylisoxazol-3-yl)-3-(4-((E)-3-(pyridin-4-yl)acrylo-
yl)phenyl)urea
[0362] 480
[0363] Compound P1 was synthesized in the manner outlined
above.
Synthesis of Chalcone Derivatives
[0364] 481482
Synthesis of Compound P2:
1-(5-tert-butylisoxazol-3-yl)-3-(4-(3-(1-methoxy-
naphthalen-4-yl)-1,2,4-oxadiazol-5-yl)phenyl)urea
[0365] 483
[0366] To a flask 5-tert-Butyl-3-isocyanato-isoxazole (242 mg, 1
eq) 4-aminobenzoic acid (159 mg, 1 eq) was added and dissolved in
toluene. The reaction was allowed to stir at 50.degree. C. for
three hours. The solvent removed and the mixture was purified by
HPLC. Yield: 188 mg (47%) 484
[0367] N-Hydroxy-4-methoxy-naphthalene-1-carboxamidine (1 g) and
4-[3-(5-tert-Butyl-isoxazol-3-yl)-ureido]-benzoic acid (1 eq) were
reflued in diglyme (5 mL) for 24 h and then purified to give 100 mg
of
1-(5-tert-Butyl-isoxazol-3-yl)-3-{4-[3-(4-methoxy-naphthalen-1-yl)-[1,2,4-
]oxadiazol-5-yl]-phenyl}-urea.
[0368] Compounds P3 through P9 were synthesized in a manner
analogous to Compound P2 using similar starting materials and
reagents. The structures are shown below in Table P:
15TABLE P NO. CHEMICAL STRUCTURE P2 485 P3 486 P4 487 P5 488 P6 489
P7 490 P8 491 P9 492
Example Q
Synthesis of Compounds Containing Heterocycloalkyl Groups
Synthesis of Compound Q1:
4-(3-methoxyphenyl)-N-(4-methoxyphenyl)piperazin-
e-1-carboxamide
[0369] 493
[0370] Commercially available isocyanides were reacted with a
secondary amine in toluene or DMA (1 ml) with triethyl amine (0.2
mL) at room temperature or 50 C overnight. The solvent was removed
and the compound purified by HPLC.
[0371] Compounds Q2 through Q27 were synthesized in a manner
analogous to Compound Q1 using similar starting materials and
reagents. The structures are shown below in Table Q:
16TABLE Q NO. CHEMICAL STRUCTURE Q1 494 Q2 495 Q3 496 Q4 497 Q5 498
Q6 499 Q7 500 Q8 501 Q9 502 Q10 503 Q11 504 Q12 505 Q13 506 Q30 507
Q31 508 Q32 509 Q17 510 Q18 511 Q19 512 Q20 513 Q21 514 Q22 515 Q23
516 Q24 517 Q25 518 Q26 519 Q27 520
Example R
Synthesis of Compounds Containing Heterocycloalkyl Groups with
Ether Linkers
Synthesis of Compound S1:
N-(4-(benzyloxy)phenyl)-4-(4-hydroxyphenyl)piper-
idine-1-carboxamide
[0372] 521
[0373] Commercially available isocyanides were reacted with a
secondary amine in toluene or DMA (1 ml) with triethyl amine (0.2
mL) at room temperature or 50 C overnight. The solvent was removed
and the compound purified by HPLC.
[0374] Compounds R.sub.2 through R.sub.9 were synthesized in a
manner analogous to Compound R.sub.1 using similar starting
materials and reagents. The structures are shown below in Table
R:
17TABLE R NO. CHEMICAL STRUCTURE R1 522 R2 523 R3 524 R4 525 R5 526
R6 527 R7 528 R8 529 R9 530
Example S
Synthesis of Carbamothioates
Synthesis of Compound S1: S-4-methoxyphenyl
N-5-tert-butylisoxazol-3-ylcar- bamothioate
[0375] 531
Synthesis of (3-tert-Butyl-isoxazol-5-yl)-thiocarbamic acid
S-(4-methoxy-phenyl)ester
[0376] 532
[0377] A mixture of 4-Methoxy-benzenethiol (0.20 g, 1 eq) and
potassium carbonate (0.47 g, 2.5 eq) in dry THF was allowed to stir
at room temperature under argon for an hour. Then the stirred
suspension was cooled to 0.degree. C. and to it was added drop wise
a solution of phosgene (0.17 g 1.2 eq). The reaction stirred at
0.degree. C. for half an hour. Then 3-tert-Butyl-isoxazol-5-ylamine
(0.20 g, 1 eq) in THF was added dropwise. The reaction was allowed
to warm to room temperature and stirred overnight. The solvent was
removed and extracted with ethyl acetate and water. The organic
layer was dried over magnesium sulfate and solvent removed. It was
purified by HPLC. Yield: 157 mg (36%), LC/MS [MH+] 307.
[0378] Compounds S2 through S3 were synthesized in a manner
analogous to Compound S1 using similar starting materials and
reagents. The structures are shown below in Table S:
18TABLE S NO. CHEMICAL STRUCTURE S1 533 S2 534 S3 535
Example T
Synthesis of Carbamothioates
Synthesis of Compound T1: S-4-methoxyphenyl
N-5-tert-butylisoxazol-3-ylcar- bamothioate
[0379] 536
Synthesis of (3-tert-Butyl-isoxazol-5-yl)-thiocarbamic acid
S-(4-methoxy-phenyl)ester
[0380] 537
[0381] A mixture of 4-Methoxy-benzenethiol (0.20 g, 1 eq) and
potassium carbonate (0.47 g, 2.5 eq) in dry THF was allowed to stir
at room temperature under argon for an hour. Then the stirred
suspension was cooled to 0.degree. C. and to it was added drop wise
a solution of phosgene (0.17 g 1.2 eq). The reaction stirred at
0.degree. C. for half an hour. Then 3-tert-Butyl-isoxazol-5-ylamine
(0.20 g, 1 eq) in THF was added dropwise. The reaction was allowed
to warm to room temperature and stirred overnight. The solvent was
removed and extracted with ethyl acetate and water. The organic
layer was dried over magnesium sulfate and solvent removed. It was
purified by HPLC. Yield: 157 mg (36%), LC/MS [MH.sup.+] 307.
Example V
Synthesis of Ureas
[0382] Synthesis of Compound V1: 538
[0383] 1 gm. (6 mmol) of 5-tert-butyl-isoxazole-3-isocyanate and
0.83 gm (6 mmol) 4-nitro-phenylamine were dissolved in 20 ml dry
toluene and stirred at 80.degree. C. for 24 h. The resulting
suspension was cooled to room temperature and filtered off to give
the title compound as a yellow solid. The product was used in the
next step without further purification. Yield: 1.7 g (92%), LC/MS
[MH.sup.+] 305. 539
[0384] 1.5 gm of
1-(5-tert-butyl-isoxazol-3-yl)-3-(4-nitro-phenyl)-urea was
dissolved in 50 ml THF and 0.1 g of 10% Pd/C was added. The
solution was stirred under hydrogen at 50 psi. for 24 h than
filtered trough Celite pad. The organic solvent was evaporated
under vacuum and the resulting residue was triturated with ethyl
acetate. Yield: 1.3 g (96%), LC/MS [MH.sup.+] 275. 540
[0385] 1 equivalent of the substituted sulfonyl chloride and 1
equivalent of the substituted aniline in DMF were added, and the
solution stirred at room temperature for 8 h. The final product was
purified by preparative HPLC.
[0386] Compounds V2 through V4 were synthesized in a manner
analogous to Compound V1 using similar starting materials and
reagents. The structures are shown below in Table V:
19TABLE V NO. CHEMICAL STRUCTURE V1 541 V2 542 V3 543 V4 544
Example W
[0387] Compounds W1 and W2 were made by procedures know in the art
or described herein.
20TABLE W NO. CHEMICAL STRUCTURE W1 545 W2 546
Example Z
Commercially Available Ureas
[0388] Compounds Z1-Z93 as shown in Table Z are commercially
available:
21TABLE Z NO. CHEMICAL STRUCTURE Z 1 547 Z 2 548 Z 3 549 Z 4 550 Z
5 551 Z 6 552 Z 7 553 Z 8 554 Z 9 555 Z10 556 Z11 557 Z12 558 Z13
559 Z14 560 Z15 561 Z16 562 Z17 563 Z18 564 Z19 565 Z20 566 Z21 567
Z22 568 Z23 569 Z24 570 Z25 571 Z26 572 Z27 573 Z28 574 Z29 575 Z30
576 Z31 577 Z32 578 Z33 579 Z34 580 Z35 581 Z36 582 Z38 583 Z39 584
Z40 585 Z43 586 Z44 587 Z45 588 Z46 589 Z47 590 Z48 591 Z49 592 Z50
593 Z51 594 Z52 595 Z53 596 Z54 597 Z55 598 Z56 599 Z57 600 Z58 601
Z59 602 Z60 603 Z61 604 Z62 605 Z63 606 Z64 607 Z65 608 Z66 609 Z67
610 Z68 611 Z69 612 Z70 613 Z71 614 Z72 615 Z73 616 Z74 617 Z75 618
Z76 619 Z77 620 Z78 621 Z79 622 Z80 623 Z81 624 Z82 625 Z83 626 Z84
627 Z85 628 Z86 629 Z87 630 Z88 631 Z89 632 Z90 633 Z91 634 Z92 635
Z93 636
Binding Constant (K.sub.d) Measurements for Small-Molecule-Kinase
Interactions
[0389] Methods for measuring binding affinities for interactions
between small molecules and kinases including FLT3, c-KIT, p38,
STK-10, MKNK2, ABL(T334I) [a.k.a. ABL(T315I)], VEGFR2 (a.k.a. KDR),
and EGFR are described in detail in U.S. application Ser. No.
10/873,835, which is incorporated by reference herein in its
entirety. The components of the assays include human kinases
expressed as fusions to T7 bacteriophage particles and immobilized
ligands that bind to the ATP site of the kinases. For the assay,
phage-displayed kinases and immobilized ATP site ligands are
combined with the compound to be tested. If the test compound binds
the kinase it competes with the immobilized ligand and prevents
binding to the solid support. If the compound does not bind the
kinase, phage-displayed proteins are free to bind to the solid
support through the interaction between the kinase and the
immobilized ligand. The results are read out by quantitating the
amount of fusion protein bound to the solid support, which is
accomplished by either traditional phage plaque assays or by
quantitative PCR (qPCR) using the phage genome as a template. To
determine the affinity of the interactions between a test molecule
and a kinase, the amount of phage-displayed kinase bound to the
solid support is quantitated as a function of test compound
concentration. The concentration of test molecule that reduces the
number of phage bound to the solid support by 50% is equal to the
K.sub.d for the interaction between the kinase and the test
molecule. Typically, data are collected for twelve concentrations
of test compound and, the resultant binding curve is fit to a
non-cooperative binding isotherm to calculate K.sub.d.
[0390] Described in the exemplary assays below is data from binding
with varying kinases. Binding values are reported as follows "+"
for representative compounds exhibiting a binding dissociation
constant (Kd) of 10,000 nM or higher; "++" for representative
compounds exhibiting a Kd of 1,000 nM to 10,000 nM; "+++" for
representative compounds exhibiting a Kd of 100 nM to 1,000 nM; and
"++++" for representative compounds exhibiting a Kd of less than
100 nM. The term "ND" represents non-determined values.
The Affinity of the Compounds for FLT3
[0391] The ability of FLT3 kinase inhibitors to inhibit cellular
proliferation was also examined. MV4:11 was a cell line derived
from a patient with acute myelogenous leukemia. It expressed a
mutant FLT3 protein that was constitutively active. MV4:11 cells
were grown in the presence of candidate FLT3 inhibitor molecules,
resulting in significantly decreased proliferation of the
leukemia-derived cells in the presence of compound. Inhibition of
FLT3 kinase activity prevented proliferation of these cells, and
thus the MV4:11 cell line can be used a model for cellular activity
of small molecule inhibitors of FLT3.
[0392] FLT3 Assay Using MV4,11 Cells
[0393] MV4,11 cells were grown in an incubator @ 37.degree. C. in
5% CO.sub.2 in Medium 2 (RPMI, 10% FBS, 4 mM glutamine,
Penn/Strep). The cells were counted daily and the cell density was
kept between 1e5 and 8e5 cells/ml.
[0394] Day One: Enough cells were harvested for experiments to be
conducted in 50 ml conical tubes. The harvested cells were spun at
500 g for 5 min at 4.degree. C., the supernatant was then aspirated
and the cells were resuspended in the starting volume of
1.times.PBS. The cells were again spun at 500 g for 5 min at
4.degree. C. and the supernatant again aspirated. The cells were
then resuspended in medium 3 (DMEM w/ glut, 10% FBS, Penn/Strep) to
a density of 4e5 cells/ml and incubated (37.degree. C. in 5%
CO.sub.2 O/N.
[0395] Day Two: The cells were counted and enough medium 3 was
added to decrease density to 2e5 cells/ml. 50 ul (10,000 cells) was
aliquoted into each well of a 96 well optical plate using
multichannel pipetman. The compound plate was then set up by
aliquoting 3 .mu.l of negative control (DMSO) into column 1 of a 96
well 300 ul polypropylene plate, aliquoting 3 .mu.t of positive
control (10 mM AB20121) into column 12 of plate, and aliquoting 3
.mu.l of appropriate compounds from serial dilutions into columns
2-11. To each well, 150 .mu.l of Medium 3 was added and 50 .mu.l of
compound/medium mixture from compound plate into rows of optical
plate in duplicate. The cells were then incubated @ 37.degree. C.
in 5% CO.sub.2 for 3 days.
[0396] Day Five: MTS was thawed in a H.sub.2O bath. 20 .mu.l of MTS
was added to each well of optical plate and the cells were
incubated (37.degree. C. in 5% CO.sub.2 for 2 hours. The plate was
then placed on a plate shaker for 30 seconds on high speed.
[0397] Data for some of the compounds is provided below:
22 Binding Compound Assay: No. FLT3(DKIN) A33 + + + + A31 + + + A29
+ + + + A27 + + + A34 + + + A32 + + + A30 + + + + A1 + + + A39 + +
+ A37 + + + + H11 + + + + H14 + + + + H12 + + + H15 + + + + A47 + +
+ A48 + + + A46 + + + B3 + + + + A54 + + + Z57 + + + Z49 + + + Z76
+ + + Z74 + + + W1 + + + + E2 + + + E4 + + + A23 + + + + A24 + + +
+ A18 + + + J10 + + + D1 + + A11 + + A25 + + F1 + A13 + + A12 + +
N94 + + + + N93 + + + V2 + + N98 + + + V4 + V3 + J13 + + + J14 + +
+ O9 + + + + C3 + + N102 + Binding Cell Compound Assay: Assay: No.
FLT3(DKIN) CS0001:IC50 A2 + + + + ND E5 + + ND J8 + ND K1 + + + ND
G32 + + + + + + + + G33 + + + + + + + Z19 + + + ND D12 + + + ND D11
+ + + ND G28 + + + + ND H2 + + + + + + + + N85 + + + + + + + N86 +
+ + + ND N87 + + + ND N88 + + + ND N89 + + + ND N90 + + + + + + +
N101 + + + + + + + G7 + + + + + + + G4 + + + + + G6 + + + + + + +
G5 + + + + + + + N153 + + + + + + + N156 + + + + + + + N150 + + + +
G53 + + + + + G42 + + + + + G40 + + + + + G38 + + + + + + + N148 +
+ + + + N146 + + + + + + Z56 + + + + Z54 + + + + Z58 + + + Z36 + +
+ N144 + + + + + + + N142 + + + + + + + N145 + + + + + + + Binding
Cell Cell Compound Assay: Assay: Assay: No. FLT3(DKIN) CS0001:IC50
CS0005:IC50 Z71 + + + + + + + ND Z72 + + + ND ND D2 + + + + ND ND
A14 + + + + ND ND A7 + + + ND ND G31 + + + + + + + + + + + G14 + +
+ + + + + + + + + G13 + + + + + + + + ND G15 + + + + + + + + ND G17
+ + + + ND ND G19 + + + + ND ND G21 + + + + + + + + ND N25 + + + +
+ + + ND N28 + + + + + + + + + + + + N30 + + + + ND N27 + + + + ND
N29 + + + + + + ND Binding Cell Cell Cell Compound Assay: Assay:
Assay: Assay: No. 4 FLT3(DKIN) CS0001:IC50 CS0005:IC50 CS0002:IC50
G34 + + + + + + + + + + + + G35 + + + + + + + + + + + + D10 + + +
ND ND ND H4 + + + ND ND ND H7 + + ND ND ND H8 + + + ND ND ND H5 + +
+ + ND ND ND G30 + + + + + + + + ND + + C2 + + ND ND ND G37 + + + +
+ + + + + + + + + A15 + + + + + + + + + + + + + A3 + + + + + + + +
+ + + + + + N91 + + + + ND ND ND V1 + + ND ND ND N92 + + + + + + +
+ + + + + + Z45 + + ND ND ND N95 + + + ND ND ND N96 + + + + + + + +
ND ND N97 + + + ND ND ND G39 + + + + + + + + + + + + + N100 + + + +
+ + + + ND + P7 + + + + + + + + + + + + P8 + + + ND ND ND N84 + + +
+ + + + + ND ND G10 + + + + + + + + + + + + + G11 + + + + + + + + +
+ + ND G12 + + + + + + + + + + + + + + G16 + + + + + + + + + + + +
+ G18 + + + + + + + ND ND G20 + + + + + + + + ND ND G3 + + + + + +
+ + + + + + + + G55 + + + + + + + ND ND G9 + + + + + + + ND ND G8 +
+ + + + ND ND G56 + + + + + + + + + + + + ND P9 + + + + + + + ND ND
P6 + + + + + ND ND N24 + + + + + ND ND N26 + + + + + + + ND N23 + +
+ + + + + + ND + + N32 + + + + + + + ND + N34 + + + + + + + + + + +
+ ND N31 + + + + + ND ND N33 + + + + ND ND N35 + + + + + ND ND N37
+ + + + + ND ND N151 + + + + + + + + + + + + + N149 + + + + + ND ND
N147 + + + + + + + + + + + + + + G48 + + + + + ND ND G46 + + + + ND
ND G44 + + + + ND ND N143 + + + + + + + + + + + + + + N141 + + + +
+ + + ND ND N139 + + + + + + ND ND N138 + + + + + + ND + G51 + + +
+ + + + ND ND N137 + + + + + + + + + + + + + G49 + + + + + ND ND
G22 + + + + + + + + + + + + + G25 + + + + + + + + + + + + + G24 + +
+ + + + + ND ND N136 + + + + + + + + + + + + + G27 + + + + + + + ND
ND N121 + + + + + + + + + + + + + + N119 + + + + + + + + + + + + +
+ N117 + + + + + + + + ND ND N115 + + + + + + + + + + + + + + +
N113 + + + + + + + + + + + + + N109 + + + + + + + + + + + + N105 +
+ + + + + + ND ND N173 + + + + + + + + + + + + + N126 + + + + + + +
+ + + + + + + + N122 + + + + + + + + + + + + + N120 + + + + + + +
ND ND N116 + + + + + + + + + + + + + + N114 + + + + + + + + + + + +
+ N100 + + + + + + + + + + + + + + N6 + + + + + ND ND N3 + + + + +
+ + + + + + + + + N5 + + + + + + + + + + + + + + + N8 + + + + + + +
+ + + + + + N10 + + + + + + + + + + ND N7 + + + + + + + + + + + + +
N9 + + + + + + + + + + + + + + N12 + + + + + + + ND ND N14 + + + +
+ + + ND ND N11 + + + + ND ND N13 + + + + + + + + ND ND N16 + + + +
+ + + + + + + + N18 + + + + + ND ND N15 + + + + + ND ND N17 + + + +
+ + + + ND + + N19 + + + + ND ND N21 + + + + + + + + + + + ND N20 +
+ + + + + + ND ND N22 + + + + + + + ND ND P3 + + + + + + ND ND P5 +
+ + + + + + + + + + + P4 + + + + + + + + ND ND N83 + + + + + + + ND
ND N39 + + + + + + ND ND N41 + + + + + + + + + + + + + G26 + + + +
+ + + + + + + + + + N40 + + + + + + + + + + + + + N42 + + + + + ND
ND N44 + + + + ND ND O10 + + + + + + + ND ND N43 + + + + + + + ND
ND N45 + + + + + + + ND ND N48 + + + + + + + + ND + + N50 + + + + +
+ + + + + + + + N47 + + + + + + + ND + + + N49 + + + + + + + + ND +
+ + N54 + + + + + + + + + + + + + + + N51 + + + + + + + + + + + + +
+ N53 + + + + + + + ND + + N56 + + + + + + + ND ND N58 + + + + + +
ND ND N55 + + + + + + + ND ND N57 + + + + + + + + + + + + + A20 + +
+ + + + ND ND O6 + + + + + + + + + + + + + O5 + + + + + + + + + + +
+ + + N60 + + + + + + + + + + + + + N62 + + + + + + + ND ND N64 + +
+ + + + + ND + Binding Cell Cell Cell Binding Compound Assay: Asay:
Assay: Assay: Assay: No. FLT3(DKIN) CS0001:IC50 CS0005:IC50
CS0002:IC50 FLT3(JMminus) N66 + + + + + + + ND ND ND N59 + + + + +
+ + ND + ND N61 + + + + + + ND ND ND N65 + + + + + + + + + + + + ND
N68 + + + + + + + ND + + + ND N70 ND + + ND ND + + + N69 ND + + +
ND ND + + N71 ND + + ND ND + + + N73 ND + + + + + + + + + + + + + +
+ N72 ND + + + + + + + + + + + + + + N74 ND + + + + + + + + + + + +
+ + N76 ND + + + ND ND + + + N75 ND + ND + + + + + + N80 ND + + +
ND ND + + + N78 ND + + + ND ND + + + N79 ND + + + + + + + + + + + +
+ + N81 ND + + + + + + + + + + + + + A21 ND + + + ND + + + + + N82
ND + + + ND + + + + + O8 ND + + + ND + + + + + + O7 ND + + + + ND +
+ + + + N169 ND + + + ND ND + + + N171 ND + + + ND ND + + N170 ND +
+ + ND ND + + + + N172 ND + + ND ND + + + N166 ND + + + ND + + + +
+ + N159 ND + + + + + + + + + + + + + + N161 ND + + + + + + + + + +
+ + + + N127 ND + + + + + + + + + + + + + + N157 ND + + + + + + + +
+ + + + + N162 ND + + + + + + + + + + + + + + N164 ND + + + + ND +
+ + + + + N163 ND + + + + + + + + + + + + + + N160 ND + + + + + + +
+ + + + + + N165 ND + + + + ND + + + + A57 ND + + + + ND + + + +
N128 ND + + ND ND ND N129 ND + + + ND ND + + + N174 ND + + ND ND +
+ + N176 ND + ND ND + + + N177 ND + + ND ND + + + N178 ND + + + ND
+ + + + + + N179 ND + + + ND ND + + + N180 ND + + + ND ND + + + +
N181 ND + + + + ND + + + + + + N182 ND + + ND ND + + + N183 ND + +
ND ND + + + + N184 ND + + + + ND + + + + + + N185 ND + + ND ND + +
+ A55 ND + ND ND + + A56 ND + + + ND + + + + Binding Cell Cell Cell
Binding Binding Compound Assay: Asay: Assay: Assay: Assay: Assay
No. FLT3(DKIN) CS0001:IC50 CS0005:IC50 CS0002:IC50 FLT3(JMminus)
FLT3(JMplus) N188 + + + + + + + + + + + + + + + + + + + + N1 + + +
+ + + + ND ND ND ND O2 + + + + + + + + + + + + + ND ND A53 + + + +
ND ND ND ND N133 + + + + + + + + + + + + + + ND ND N131 + + + + + +
+ + + + + + ND ND N134 + + + + + + + + + + + + + + + + + + + + +
N132 + + + + + + ND ND ND ND G1 + + + + + + + + + + + + + ND ND
N108 + + + + + + + + + + + + + + ND ND N106 + + + + + + + + + + + +
+ + + + + + + + N174 + + + + + + + ND ND ND ND O4 + + + + + + + + +
+ + + + + ND ND O1 + + + + + + + + + + + + + + ND ND O3 + + + + + +
+ + + + + + + + ND ND N4 + + + + + + + + + + + + + + ND ND Binding
Cell Compound Assay: Assay: No. FLT3(DKIN) CS0001:IC50 637 + + + +
+ + + Binding Assay: Cell Cell Cell Binding Compound FLT3 Asay:
Assay: Assay: Assay: No. (DKIN) CS0001:IC50 CS0005:IC50 CS0002:IC50
FLT3(JMminus) 638 ND + + + ND + + + + + 639 ND + + + ND + + + +
+
[0398] All references cited herein, including patents, patent
applications, and publications, are herby incorporated by reference
in their entireties, whether previously specifically incorporated
or not.
[0399] Having now fully described this invention, it will be
appreciated by those skilled in the art that the same can be
performed within a wide range of equivalent parameters,
concentrations, and conditions without departing from the spirit
and scope of the invention and without undue experimentation.
[0400] While this invention has been described in connection with
specific embodiments thereof, it will be understood that it is
capable of further modifications. This application is intended to
cover any variations, uses, or adaptations of the invention
following, in general, the principles of the invention and
including such departures from the present disclosure as come
within known or customary practice within the art to which the
invention pertains and as may be applied to the essential features
hereinbefore set forth.
* * * * *