U.S. patent application number 11/746543 was filed with the patent office on 2008-03-27 for methods for treating blood disorders.
Invention is credited to Susan P. Perrine.
Application Number | 20080075692 11/746543 |
Document ID | / |
Family ID | 38626483 |
Filed Date | 2008-03-27 |
United States Patent
Application |
20080075692 |
Kind Code |
A1 |
Perrine; Susan P. |
March 27, 2008 |
METHODS FOR TREATING BLOOD DISORDERS
Abstract
Methods of treating blood disorders are described.
Inventors: |
Perrine; Susan P.; (Newton,
MA) |
Correspondence
Address: |
SWANSON & BRATSCHUN, L.L.C.
8210 SOUTHPARK TERRACE
LITTLETON
CO
80120
US
|
Family ID: |
38626483 |
Appl. No.: |
11/746543 |
Filed: |
May 9, 2007 |
Related U.S. Patent Documents
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Application
Number |
Filing Date |
Patent Number |
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60799054 |
May 9, 2006 |
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Current U.S.
Class: |
424/85.2 ;
514/224.2; 514/266.4; 514/311; 514/345; 514/348; 514/361; 514/369;
514/384; 514/386; 514/438; 514/448; 514/457; 514/464; 514/469;
514/557; 514/568; 514/570; 514/574; 514/709; 546/174; 548/183;
549/447; 562/495 |
Current CPC
Class: |
A61K 31/415 20130101;
A61K 31/36 20130101; A61K 31/39 20130101; A61K 31/194 20130101;
A61K 31/353 20130101; A61K 31/192 20130101; C07D 311/12 20130101;
A61K 31/433 20130101; A61P 7/00 20180101; A61P 43/00 20180101; A61P
7/06 20180101; A61K 31/517 20130101; C07D 285/125 20130101 |
Class at
Publication: |
424/085.2 ;
514/224.2; 514/266.4; 514/311; 514/345; 514/348; 514/361; 514/369;
514/384; 514/386; 514/438; 514/448; 514/457; 514/464; 514/469;
514/557; 514/568; 514/570; 514/574; 514/709; 546/174; 548/183;
549/447; 562/495 |
International
Class: |
A61K 38/20 20060101
A61K038/20; A61K 31/10 20060101 A61K031/10; A61K 31/19 20060101
A61K031/19; A61K 31/34 20060101 A61K031/34; A61K 31/35 20060101
A61K031/35; A61K 31/36 20060101 A61K031/36; A61K 31/38 20060101
A61K031/38; A61K 31/41 20060101 A61K031/41; C07C 63/64 20060101
C07C063/64; C07D 317/44 20060101 C07D317/44; C07D 277/04 20060101
C07D277/04; C07D 215/12 20060101 C07D215/12; A61P 7/00 20060101
A61P007/00; A61K 31/415 20060101 A61K031/415; A61K 31/425 20060101
A61K031/425; A61K 31/44 20060101 A61K031/44; A61K 31/47 20060101
A61K031/47; A61K 31/517 20060101 A61K031/517; A61K 31/54 20060101
A61K031/54 |
Claims
1. A method for treating or preventing a blood disorder in a
subject, comprising administering to said subject an effective
amount of a compound of formula I: ##STR85## wherein R.sup.1 is
hydroxy or alkoxy; X is C(O), C(S), SO, SO.sub.2 or PO.sub.2;
R.sup.2 and R.sup.3 are each independently hydrogen, alkyl,
halogen, hydroxyl, thiol, alkenyl, alkynyl, aryl, acyl, alkoxy,
amino, alkylamino or heterocyclic; R.sup.4 is alkyl, cycloalkyl,
alkenyl, alkynyl, arylalkyl, amido, alkylamino, amino, arylamino,
alkylcarbonyl, arylcarbonyl, alkylaminocarbonyl, alkoxy,
alkoxycarbonyl, alkylcarbonyloxy, alkyloxycarbonyloxy,
arylcarbonyloxy, aryloxy, thiol, alkylthio, arylthio, alkenyl,
heterocyclic, heteroaryl, halogen or ##STR86## R.sup.5 is hydrogen,
alkyl, alkenyl, alkynyl, aryl, arylalkyl, amido, alkylamino, amino,
arylamino, alkylcarbonyl, arylcarbonyl, alkylaminocarbonyl, alkoxy,
alkoxycarbonyl, alkylcarbonyloxy, alkyloxycarbonyloxy,
arylcarbonyloxy, aryloxy, thiol, alkylthio, arylthio, alkenyl,
heterocyclic, hydroxy, nitro, halogen or optionally linked to
R.sup.6 to form a ring; R.sup.6 is hydrogen, alkyl, alkenyl,
alkynyl, aryl, arylalkyl, amido, alkylamino, amino, arylamino,
alkylcarbonyl, arylcarbonyl, alkylaminocarbonyl, alkoxy,
alkoxycarbonyl, alkylcarbonyloxy, alkyloxycarbonyloxy,
arylcarbonyloxy, aryloxy, thiol, alkylthio, arylthio, alkenyl,
heterocyclic, hydroxy, nitro, halogen or optionally linked to
R.sup.5 or R.sup.7 to form a ring; R.sup.7 is hydrogen, alkyl,
alkenyl, alkynyl, aryl, arylalkyl, amido, alkylamino, amino,
arylamino, alkylcarbonyl, arylcarbonyl, alkylaminocarbonyl, alkoxy,
alkoxycarbonyl, alkylcarbonyloxy, alkyloxycarbonyloxy,
arylcarbonyloxy, aryloxy, thiol, alkylthio, arylthio, alkenyl,
heterocyclic, hydroxy, nitro, halogen or optionally linked to
R.sup.6 or R.sup.8 to form a ring; R.sup.8 is hydrogen, alkyl,
alkenyl, alkynyl, aryl, arylalkyl, amido, alkylamino, amino,
arylamino, alkylcarbonyl, arylcarbonyl, alkylaminocarbonyl, alkoxy,
alkoxycarbonyl, alkylcarbonyloxy, alkyloxycarbonyloxy,
arylcarbonyloxy, aryloxy, thiol, alkylthio, arylthio, alkenyl,
heterocyclic, hydroxy, nitro, halogen or optionally linked to
R.sup.7 or R.sup.9 to form a ring; R.sup.9 is hydrogen, alkyl,
alkenyl, alkynyl, aryl, arylalkyl, amido, alkylamino, amino,
arylamino, alkylcarbonyl, arylcarbonyl, alkylaminocarbonyl, alkoxy,
alkoxycarbonyl, alkylcarbonyloxy, alkyloxycarbonyloxy,
arylcarbonyloxy, aryloxy, thiol, alkylthio, arylthio, alkenyl,
heterocyclic, hydroxy, nitro, halogen or optionally linked to
R.sup.8 to form a ring; and racemates, isolated enantiomers or
diastereomers, and pharmaceutically acceptable salts thereof;
provided that when R.sup.4 is ##STR87## R.sup.5, R.sup.6, R.sup.7,
R.sup.8, and R.sup.9 are each not hydrogen; and when R.sup.6,
R.sup.7, R.sup.8, and R.sup.9 are each hydrogen, R.sup.5 is not
methoxy; and when R.sup.5, R.sup.7, R.sup.8, R.sup.9 are hydrogen,
R.sup.6 is not methoxy; and when R.sup.5, R.sup.8 and R.sup.9 are
hydrogen, R.sup.6 and R.sup.7 are not methoxy.
2. The method of claim 1, wherein R.sup.1 is hydroxy, X is C(O),
R.sup.4 is ##STR88## and R.sup.2, R.sup.3 and R.sup.5 are each
hydrogen.
3. The method of claim 2, wherein R.sup.6, R.sup.8 and R.sup.9 are
each hydrogen, and R.sup.7 is alkoxy.
4. The method of claim 3, wherein R.sup.7 is methoxy.
5. The method of claim 2, wherein R.sup.7 and R.sup.8 are each
hydrogen, and R.sup.6 and R.sup.9 are each alkyl.
6. The method of claim 5, wherein R.sup.6 and R.sup.9 are
methyl.
7. The method of claim 2, wherein R.sup.8 are R.sup.9 are each
hydrogen and R.sup.6 and R.sup.7 are each hydroxyl.
8. The method of claim 2, wherein R.sup.8 are R.sup.9 are each
hydrogen and R.sup.6 and R.sup.7 are linked by --O--CH.sub.2--O--
to form a ring.
9. The method of claim 2, wherein R.sup.6 and R.sup.9 are each
hydrogen, R.sup.7 is alkoxy and R.sup.8 is hydroxyl.
10. The method of claim 1, wherein R.sup.4 is heteroaryl.
11. The method of claim 10, wherein R.sup.4 is quinoline or
substituted or unsubstituted thiophene.
12. The method of claim 11, wherein R.sup.4 is chlorothiophene.
13. The method of claim 1, wherein said compound is: ##STR89##
14. A method for treating or preventing a blood disorder in a
subject, comprising administering to said subject an effective
amount of a compound of formula II: ##STR90## wherein R.sup.1' is
hydroxy or alkoxy; Y is C(O); n is 0 or an integer from 1 to 5;
R.sup.10 and R.sup.10' are each independently hydrogen, alkyl,
halogen, hydroxyl, thiol, alkenyl, alkynyl, aryl, acyl, alkoxy,
amino, alkylamino, heterocyclic or optionally joined to form a
ring; R.sup.11 is CR.sup.11'R.sup.11''R.sup.11''', alkenyl,
cycloalkyl, alkynyl, arylalkyl, amido, alkylamino, amino,
arylamino, carbonylalkyl, alkylcarbonyl, arylcarbonyl,
alkylaminocarbonyl, alkoxycarbonyl, alkylcarbonyloxy,
alkyloxycarbonyloxy, arylcarbonyloxy, thiol, alkylthio, arylthio,
alkenyl, heterocyclic, heteroaryl, hydroxy, halogen or ##STR91##
R.sup.11' and R.sup.11'' are each independently hydrogen, alkyl,
alkenyl, alkynyl, aryl, arylalkyl, amido, alkylamino, amino,
arylamino, alkylcarbonyl, arylcarbonyl, alkylaminocarbonyl, alkoxy,
alkoxycarbonyl, alkylcarbonyloxy, alkyloxycarbonyloxy,
arylcarbonyloxy, aryloxy, thiol, alkylthio, arylthio, alkenyl,
heterocyclic, hydroxyl, halogen or R.sup.11' and R.sup.11'' are
optionally joined to form a ring; R.sup.11''' is alkenyl, alkynyl,
aryl, arylalkyl, amido, alkylamino, amino, arylamino,
alkylcarbonyl, arylcarbonyl, alkylaminocarbonyl, alkoxy,
alkoxycarbonyl, alkylcarbonyloxy, alkyloxycarbonyloxy,
arylcarbonyloxy, aryloxy, thiol, alkylthio, arylthio, alkenyl,
heterocyclic, hydroxy or halogen; R.sup.12 is hydrogen, alkenyl,
alkynyl, aryl, arylalkyl, amido, alkylamino, amino, arylamino,
alkylcarbonyl, arylcarbonyl, alkylaminocarbonyl, alkoxy,
alkoxycarbonyl, alkylcarbonyloxy, alkyloxycarbonyloxy,
arylcarbonyloxy, aryloxy, thiol, alkylthio, arylthio, alkenyl,
heterocyclic or optionally linked to R.sup.13 to form a ring;
R.sup.13 is hydrogen, alkyl, alkenyl, alkynyl, aryl, arylalkyl,
amido, alkylamino, amino, arylamino, alkylcarbonyl, arylcarbonyl,
alkylaminocarbonyl, alkoxy, alkoxycarbonyl, alkylcarbonyloxy,
alkyloxycarbonyloxy, arylcarbonyloxy, aryloxy, thiol, alkylthio,
arylthio, alkenyl, heterocyclic, hydroxy or optionally linked to
R.sup.12 or R.sup.14 to form a ring; R.sup.14 is hydrogen, alkenyl,
alkynyl, aryl, arylalkyl, amido, alkylamino, amino, arylamino,
alkylcarbonyl, arylcarbonyl, alkylaminocarbonyl, alkoxy,
alkoxycarbonyl, alkylcarbonyloxy, alkyloxycarbonyloxy,
arylcarbonyloxy, aryloxy, thiol, alkylthio, arylthio, alkenyl,
heterocyclic, halogen or optionally linked to R.sup.13 or R.sup.15
to form a ring; R.sup.15 is hydrogen, alkenyl, alkynyl, aryl,
arylalkyl, amido, alkylamino, amino, arylamino, alkylcarbonyl,
arylcarbonyl, alkylaminocarbonyl, alkoxy, alkoxycarbonyl,
alkylcarbonyloxy, alkyloxycarbonyloxy, arylcarbonyloxy, aryloxy,
thiol, alkylthio, arylthio, alkenyl, heterocyclic, halogen or
optionally linked to R.sup.14 or R.sup.16 to form a ring; R.sup.16
is hydrogen, alkenyl, alkynyl, aryl, arylalkyl, amido, alkylamino,
amino, arylamino, alkylcarbonyl, arylcarbonyl, alkylaminocarbonyl,
alkoxy, alkoxycarbonyl, alkylcarbonyloxy, alkyloxycarbonyloxy,
arylcarbonyloxy, aryloxy, thiol, alkylthio, arylthio, alkenyl,
heterocyclic, halogen or optionally linked to R.sup.15 to form a
ring; and racemates, isolated enantiomers or diastereomers, and
pharmaceutically acceptable salts thereof; provided that when
R.sup.11 is ##STR92## R.sup.12, R.sup.13, R.sup.14, R.sup.15, and
R.sup.16 are each not hydrogen; and provided when n is 2, R.sup.11
is ##STR93## and R.sup.10, R.sup.10', R.sup.12, R.sup.15, and
R.sup.16 are hydrogen, then R.sup.14 and R.sup.15 are not methoxy;
and provided when n is 1, R.sup.11 is ##STR94## and R.sup.10,
R.sup.10', R.sup.13, R.sup.14, and R.sup.16 are hydrogen, then
R.sup.12 and R.sup.15 are not methoxy.
15. The method of claim 14, wherein R.sup.1' is hydroxy.
16. The method of claim 15, wherein n is 5, R.sup.10 and R.sup.10'
are hydrogen and R.sup.11 is alkylcarbonyl.
17. The method of claim 15, wherein n is 2, R.sup.10 and R.sup.10'
are each hydrogen and R.sup.11 is
CR.sup.11'R.sup.11''R.sup.11'''.
18. The method of claim 17, wherein R.sup.11' and R.sup.11'' are
joined by --(CH.sub.2).sub.5-- to form a cyclohexyl ring and
R.sup.11''' is a substituted or unsubstituted heterocycle.
19. The method of claim 18, wherein R.sup.11''' is
chlorothiophene.
20. The method of claim 15, wherein n is 0.
21. The method of claim 20, wherein R.sup.11 is ##STR95##
22. The method of claim 21, wherein R.sup.13, R.sup.14, R.sup.15,
and R.sup.16 are hydrogen.
23. The method of claim 22, wherein R.sup.12 is arylthioalkyl.
24. The method of claim 22, wherein R.sup.12 is alkoxy substituted
aryloxy.
25. The method of claim 21, wherein R.sup.12, R.sup.14, R.sup.15
and R.sup.16 are hydrogen.
26. The method of claim 25, wherein R.sup.13 is a substituted or
unsubstituted heterocycle.
27. The method of claim 26, wherein R.sup.13 is chromen-2-one,
nitro-substituted pyrazole, or chloro-substituted pyrazole.
28. The method of claim 21, wherein R.sup.12, R.sup.13, R.sup.15
and R.sup.16 are hydrogen and R.sup.13 is alkoxy.
29. The method of claim 28, wherein R.sup.13 is ethoxy.
30. The method of claim 21, wherein R.sup.12, R.sup.15 and R.sup.16
are hydrogen, and R.sup.13 and R.sup.14 are linked by
--N(H)C(O)CH.sub.2S-- to form a ring.
31. The method of claim 21, wherein R.sup.14, R.sup.15 and R.sup.16
are each hydrogen, and R.sup.12 and R.sup.13 are linked by
--CH.dbd.C(CH.sub.3)O-- to form a ring.
32. The method of claim 20, wherein R.sup.11 is a substituted or
unsubstituted heterocycle.
33. The method of claim 32, wherein R.sup.11 is a substituted
aryl-substituted furan.
34. The method of claim 20, wherein R.sup.11 is a substituted or
unsubstituted cycloalkyl.
35. The method of claim 34, wherein R.sup.11 is
tetrahydrobenzothiadiazole or dihydrobenzothiophenone.
36. The method of claim 15, wherein n is 1.
37. The method of claim 36, wherein R.sup.10 and R.sup.10' are
hydrogen.
38. The method of claim 37, wherein R.sup.11 is a substituted or
unsubstituted cycloalkyl.
39. The method of claim 38, wherein R.sup.11 is dimethylcyclobutane
carboxylic acid.
40. The method of claim 37, wherein R.sup.11 is ##STR96##
41. The method of claim 40, wherein R.sup.12, R.sup.13 and R.sup.16
are hydrogen, and R.sup.14 and R.sup.15 are linked by
--O--CH.sub.2--O-- to form a ring.
42. The method of claim 37, wherein R.sup.11 is a substituted or
unsubstituted heterocycle.
43. The method of claim 42, wherein R.sup.11 is substituted
thiazolidinedione, substituted pyridinone or substituted
pyrazole.
44. The method of claim 37, wherein R.sup.11 is substituted or
unsubstituted arylamino.
45. The method of claim 44, wherein R.sup.11 arylamino is
trifluorothio-substituted arylamino.
46. The method of claim 37, wherein R.sup.11 is substituted or
unsubstituted arylthio or substituted or unsubstituted heterocyclic
thio.
47. The method of claim 46, wherein R.sup.11 is
methoxyphenylthio.
48. The method of claim 46, said R.sup.11 is substituted
triazolethio, substituted thiadiazolethio or substituted
thiophenethio.
49. The method of claim 37, wherein R.sup.11 is
CR.sup.11'R.sup.11''R.sup.11'''.
50. The method of claim 49, wherein R.sup.11' is hydrogen,
R.sup.11'' is amino and R.sup.11''' is alkoxy-substituted aryl.
51. The method of claim 36, wherein R.sup.10 is hydrogen and
R.sup.10' is alkyl.
52. The method of claim 51, wherein R.sup.10' is isopropyl.
53. The method of claim 52, wherein R.sup.11 is substituted or
unsubstituted arylthio.
54. The method of claim 53, wherein R.sup.11 is alkoxy-substituted
phenylthio or alkoxy-substituted pyrimadinylthio.
55. The method of claim 51, wherein R.sup.10' is ethyl.
56. The method of claim 55, wherein R.sup.11 is
heteroarylamino.
57. The method of claim 56, wherein R.sup.11 is
quinazolinylamino.
58. The method of claim 36, wherein R.sup.10 and R.sup.10' are
linked by --(CH.sub.2).sub.5-- to form a cyclohexyl ring.
59. The method of claim 58, wherein R.sup.11 is heterocyclic
substituted carbonylalkyl.
60. The method of claim 14, wherein said compound is ##STR97##
##STR98## ##STR99##
61. A method for treating or preventing a blood disorder in a
subject, comprising administering to said subject an effective
amount of a compound of formula III: ##STR100## wherein R.sup.1''
is hydroxy or alkoxy; Z is C(S), SO, SO.sub.2 or PO.sub.2; m is 0
or an integer from 1-5; R.sup.17 and R.sup.17' are each
independently hydrogen, alkyl, halogen, hydroxyl, thiol, alkenyl,
alkynyl, aryl, acyl, alkoxy, amino, alkylamino or heterocyclic;
R.sup.18 is hydrogen, alkyl, cycloalkyl, alkenyl, alkynyl, aryl,
arylalkyl, amido, alkylamino, amino, arylamino, alkylcarbonyl,
arylcarbonyl, alkylaminocarbonyl, alkoxy, alkoxycarbonyl,
alkylcarbonyloxy, alkyloxycarbonyloxy, arylcarbonyloxy, aryloxy,
thiol, alkylthio, arylthio, alkenyl, heterocyclic, heteroaryl,
hydroxyl or halogen; and racemates, isolated enantiomers or
diastereomers, and pharmaceutically acceptable salts thereof.
62. The method of claim 61, wherein R.sup.1'' is hydroxyl, Z is
SO.sub.2, m is 0, and R.sup.18 is disubstituted aryl.
63. The method of claim 62, wherein R.sup.18 is substituted by
nitro and fluoro.
64. The method of claim 61, wherein the compound is of the formula:
##STR101##
65. A method for treating or preventing a blood disorder in a
subject, comprising administering to said subject an effective
amount of a compound of formula: ##STR102##
66. The method of any one of claims 1, 14, 61 or 65, wherein the
blood disorder is sickle cell anemia, .beta.-thalassemia,
neutropenia or thrombocytopenia.
67. The method of any one of claims 1, 14, 61 or 65, wherein said
compound stimulates fetal hemoglobin production.
68. The method of any one of claim 1, 14, 61 or 65, wherein said
compound stimulates hematopoiesis.
69. The method of any one of claims 1, 14, 61 or 65, wherein said
compound stimulates erythropoiesis.
70. The method of any one of claims 1, 14, 61 or 65, wherein said
compound stimulates myelopoiesis.
71. The method of any one of claims 1, 14, 61 or 65, wherein said
compound stimulates neutrophil production.
72. The method of any one of claims 1, 14, 61 or 65, wherein said
compound is administered in combination with one or more
cytokines.
73. The method of claim 72, wherein said cytokine is selected from
the group consisting of IL-3, GM-CSF, C-CSF, SCF and IL-6.
74. A pharmaceutical composition comprising an effective amount of
a compound of any one of claims 1, 13, 14, 60, 61 or 65 and
racemates, isolated enantiomers or diastereomers thereof, and a
pharmaceutically acceptable carrier.
75. A compound of formula I: ##STR103## wherein R.sup.1 is hydroxy
or alkoxy; X is C(O), C(S), SO, SO.sub.2 or PO.sub.2; R.sup.2 and
R.sup.3 are each independently hydrogen, alkyl, halogen, hydroxyl,
thiol, alkenyl, alkynyl, aryl, acyl, alkoxy, amino, alkylamino or
heterocyclic; R.sup.4 is alkyl, cycloalkyl, alkenyl, alkynyl,
arylalkyl, amido, alkylamino, amino, arylamino, alkylcarbonyl,
arylcarbonyl, alkylaminocarbonyl, alkoxy, alkoxycarbonyl,
alkylcarbonyloxy, alkyloxycarbonyloxy, arylcarbonyloxy, aryloxy,
thiol, alkylthio, arylthio, alkenyl, heterocyclic, heteroaryl,
halogen or ##STR104## R.sup.5 is hydrogen, alkyl, alkenyl, alkynyl,
aryl, arylalkyl, amido, alkylamino, amino, arylamino,
alkylcarbonyl, arylcarbonyl, alkylaminocarbonyl, alkoxy,
alkoxycarbonyl, alkylcarbonyloxy, alkyloxycarbonyloxy,
arylcarbonyloxy, aryloxy, thiol, alkylthio, arylthio, alkenyl,
heterocyclic, hydroxy, nitro, halogen or optionally linked to
R.sup.6 to form a ring; R.sup.6 is hydrogen, alkyl, alkenyl,
alkynyl, aryl, arylalkyl, amido, alkylamino, amino, arylamino,
alkylcarbonyl, arylcarbonyl, alkylaminocarbonyl, alkoxy,
alkoxycarbonyl, alkylcarbonyloxy, alkyloxycarbonyloxy,
arylcarbonyloxy, aryloxy, thiol, alkylthio, arylthio, alkenyl,
heterocyclic, hydroxy, nitro, halogen or optionally linked to
R.sup.5 or R.sup.7 to form a ring; R.sup.7 is hydrogen, alkyl,
alkenyl, alkynyl, aryl, arylalkyl, amido, alkylamino, amino,
arylamino, alkylcarbonyl, arylcarbonyl, alkylaminocarbonyl, alkoxy,
alkoxycarbonyl, alkylcarbonyloxy, alkyloxycarbonyloxy,
arylcarbonyloxy, aryloxy, thiol, alkylthio, arylthio, alkenyl,
heterocyclic, hydroxy, nitro, halogen or optionally linked to
R.sup.6 or R.sup.8 to form a ring; R.sup.8 is hydrogen, alkyl,
alkenyl, alkynyl, aryl, arylalkyl, amido, alkylamino, amino,
arylamino, alkylcarbonyl, arylcarbonyl, alkylaminocarbonyl, alkoxy,
alkoxycarbonyl, alkylcarbonyloxy, alkyloxycarbonyloxy,
arylcarbonyloxy, aryloxy, thiol, alkylthio, arylthio, alkenyl,
heterocyclic, hydroxy, nitro, halogen or optionally linked to
R.sup.7 or R.sup.9 to form a ring; R.sup.9 is hydrogen, alkyl,
alkenyl, alkynyl, aryl, arylalkyl, amido, alkylamino, amino,
arylamino, alkylcarbonyl, arylcarbonyl, alkylaminocarbonyl, alkoxy,
alkoxycarbonyl, alkylcarbonyloxy, alkyloxycarbonyloxy,
arylcarbonyloxy, aryloxy, thiol, alkylthio, arylthio, alkenyl,
heterocyclic, hydroxy, nitro, halogen or optionally linked to
R.sup.8 to form a ring; and racemates, isolated enantiomers or
diastereomers, and pharmaceutically acceptable salts thereof;
provided that when R.sup.4 is ##STR105## R.sup.5, R.sup.6, R.sup.7,
R.sup.8, and R.sup.9 are each not hydrogen; and when R.sup.6,
R.sup.7, R.sup.8, and R.sup.9 are each hydrogen, R.sup.5 is not
methoxy; and when R.sup.5, R.sup.7, R.sup.8, R.sup.9 are hydrogen,
R.sup.6 is not methoxy; and when R.sup.5, R.sup.8 and R.sup.9 are
hydrogen, R.sup.6 and R.sup.7 are not methoxy; and provided that
the compound is not a compound of claim 13.
76. A compound of formula II: ##STR106## wherein R.sup.1' is
hydroxy or alkoxy; Y is C(O); n is 0 or an integer from 1 to 5;
R.sup.10 and R.sup.10' are each independently hydrogen, alkyl,
halogen, hydroxyl, thiol, alkenyl, alkynyl, aryl, acyl, alkoxy,
amino, alkylamino, heterocyclic or optionally joined to form a
ring; R.sup.11 is CR.sup.11'R.sup.11''R.sup.11''', alkenyl,
cycloalkyl, alkynyl, arylalkyl, amido, alkylamino, amino,
arylamino, carbonylalkyl, alkylcarbonyl, arylcarbonyl,
alkylaminocarbonyl, alkoxycarbonyl, alkylcarbonyloxy,
alkyloxycarbonyloxy, arylcarbonyloxy, thiol, alkylthio, arylthio,
alkenyl, heterocyclic, heteroaryl, hydroxy, halogen or ##STR107##
R.sup.11' and R.sup.11'' are each independently hydrogen, alkyl,
alkenyl, alkynyl, aryl, arylalkyl, amido, alkylamino, amino,
arylamino, alkylcarbonyl, arylcarbonyl, alkylaminocarbonyl, alkoxy,
alkoxycarbonyl, alkylcarbonyloxy, alkyloxycarbonyloxy,
arylcarbonyloxy, aryloxy, thiol, alkylthio, arylthio, alkenyl,
heterocyclic, hydroxyl, halogen or R.sup.11' and R.sup.11'' are
optionally joined to form a ring; R.sup.11''' is alkenyl, alkynyl,
aryl, arylalkyl, amido, alkylamino, amino, arylamino,
alkylcarbonyl, arylcarbonyl, alkylaminocarbonyl, alkoxy,
alkoxycarbonyl, alkylcarbonyloxy, alkyloxycarbonyloxy,
arylcarbonyloxy, aryloxy, thiol, alkylthio, arylthio, alkenyl,
heterocyclic, hydroxy or halogen; R.sup.12 is hydrogen, alkenyl,
alkynyl, aryl, arylalkyl, amido, alkylamino, amino, arylamino,
alkylcarbonyl, arylcarbonyl, alkylaminocarbonyl, alkoxy,
alkoxycarbonyl, alkylcarbonyloxy, alkyloxycarbonyloxy,
arylcarbonyloxy, aryloxy, thiol, alkylthio, arylthio, alkenyl,
heterocyclic or optionally linked to R.sup.13 to form a ring;
R.sup.13 is hydrogen, alkyl, alkenyl, alkynyl, aryl, arylalkyl,
amido, alkylamino, amino, arylamino, alkylcarbonyl, arylcarbonyl,
alkylaminocarbonyl, alkoxy, alkoxycarbonyl, alkylcarbonyloxy,
alkyloxycarbonyloxy, arylcarbonyloxy, aryloxy, thiol, alkylthio,
arylthio, alkenyl, heterocyclic, hydroxy or optionally linked to
R.sup.12 or R.sup.14 to form a ring; R.sup.14 is hydrogen, alkenyl,
alkynyl, aryl, arylalkyl, amido, alkylamino, amino, arylamino,
alkylcarbonyl, arylcarbonyl, alkylaminocarbonyl, alkoxy,
alkoxycarbonyl, alkylcarbonyloxy, alkyloxycarbonyloxy,
arylcarbonyloxy, aryloxy, thiol, alkylthio, arylthio, alkenyl,
heterocyclic, halogen or optionally linked to R.sup.13 or R.sup.15
to form a ring; R.sup.15 is hydrogen, alkenyl, alkynyl, aryl,
arylalkyl, amido, alkylamino, amino, arylamino, alkylcarbonyl,
arylcarbonyl, alkylaminocarbonyl, alkoxy, alkoxycarbonyl,
alkylcarbonyloxy, alkyloxycarbonyloxy, arylcarbonyloxy, aryloxy,
thiol, alkylthio, arylthio, alkenyl, heterocyclic, halogen or
optionally linked to R.sup.14 or R.sup.16 to form a ring; R.sup.16
is hydrogen, alkenyl, alkynyl, aryl, arylalkyl, amido, alkylamino,
amino, arylamino, alkylcarbonyl, arylcarbonyl, alkylaminocarbonyl,
alkoxy, alkoxycarbonyl, alkylcarbonyloxy, alkyloxycarbonyloxy,
arylcarbonyloxy, aryloxy, thiol, alkylthio, arylthio, alkenyl,
heterocyclic, halogen or optionally linked to R.sup.15 to form a
ring; and racemates, isolated enantiomers or diastereomers, and
pharmaceutically acceptable salts thereof; provided that when
R.sup.11 is ##STR108## R.sup.12, R.sup.13, R.sup.14, R.sup.15, and
R.sup.16 are each not hydrogen; and provided when n is 2, R.sup.11
is ##STR109## and R.sup.10, R.sup.10', R.sup.12, R.sup.15, and
R.sup.16 are hydrogen, then R.sup.14 and R.sup.15 are not methoxy;
and provided when n is 1, R.sup.11 is ##STR110## and R.sup.10,
R.sup.10', R.sup.13, R.sup.14, and R.sup.16 are hydrogen, then
R.sup.12 and R.sup.15 are not methoxy; and provided that the
compound is not a compound of claim 60.
77. A compound of formula III: ##STR111## wherein R.sup.1'' is
hydroxy or alkoxy; Z is C(S), SO, SO.sub.2 or PO.sub.2; m is 0 or
an integer from 1-5; R.sup.17 and R.sup.17' are each independently
hydrogen, alkyl, halogen, hydroxyl, thiol, alkenyl, alkynyl, aryl,
acyl, alkoxy, amino, alkylamino or heterocyclic; R.sup.18 is
hydrogen, alkyl, cycloalkyl, alkenyl, alkynyl, aryl, arylalkyl,
amido, alkylamino, amino, arylamino, alkylcarbonyl, arylcarbonyl,
alkylaminocarbonyl, alkoxy, alkoxycarbonyl, alkylcarbonyloxy,
alkyloxycarbonyloxy, arylcarbonyloxy, aryloxy, thiol, alkylthio,
arylthio, alkenyl, heterocyclic, heteroaryl, hydroxyl or halogen;
and racemates, isolated enantiomers or diastereomers, and
pharmaceutically acceptable salts thereof; provided that the
compound is not a compound of claim 64.
Description
CROSS-REFERENCE TO RELATED APPLICATION
[0001] This application claims the benefit under 35 U.S.C. .sctn.
119(e) of U.S. Provisional Patent Application No. 60/799,054 filed
May 9, 2006, which provisional application is incorporated herein
by reference in its entirety.
BACKGROUND
[0002] 1. Field of the Invention
[0003] The present invention relates to methods and compounds for
the treatment of blood disorders.
[0004] 2. Description of the Related Art
[0005] The major function of red blood cells is to transport oxygen
to tissues of the body, while minor functions include the
transportation of nutrients and cytokines and the absorption of
cellular metabolites. Anemia, defined as a loss of red blood cells
or red blood cell capacity resulting in the reduction in the
ability of the blood to transport oxygen, may be chronic or acute.
Chronic anemia may be caused by extrinsic red blood cell
abnormalities, intrinsic abnormalities or impaired production of
red blood cells. Extrinsic or extra-corpuscular abnormalities
include antibody-mediated disorders such as transfusion reactions
and erythroblastosis, mechanical trauma to red cells such as
micro-angiopathic hemolytic anemias, thrombotic thrombocytopenic
purpura and disseminated intravascular coagulation. In addition,
infections by parasites such as Plasmodium, chemical injuries from,
for example, lead poisoning, and sequestration in the mononuclear
system such as by hyperspienism can result in red blood cell
disorders and deficiencies.
[0006] Impaired red blood cell production can occur by disturbing
the proliferation and differentiation of the stem cells or
committed cells. Some of the more common diseases of red cell
production include aplastic anemia, sickle cell anemia,
.beta.-thalassemia, hypoplastic anemia, pure red cell aplasia and
anemia associated with renal failure or endocrine disorders.
Disturbances of the proliferation and differentiation of
erythroblasts include defects in DNA synthesis such as impaired
utilization of vitamin B.sub.12 or folic acid and the megaloblastic
anemias, defects in heme or globin synthesis, and anemias of
unknown origins such as sideroblastic anemia, anemia associated
with chronic infections such as malaria, trypanosomiasis, HIV,
hepatitis virus or other viruses, and myelophthisic anemias caused
by marrow deficiencies.
[0007] Symptoms of anemia include feelings of weakness or fatigue,
pallor, shortness of breath, an increase in cardiac output, which
may lead to palpitations and sweatiness. In severe cases, anemia
can lead to death by heart failure. Current treatments for anemia
depend on the type of anemia the patient suffers from. Monitoring
of the diet to increase iron intake may be prescribed, as well as
iron supplementation. In some cases, medication or blood
transfusions may be necessary.
[0008] Sickle cell disease and .beta.-thalassemia are two of the
most common genetic disorders in the word. These disorders are
caused by molecular mutations affecting the .beta.-globin genes for
adult hemoglobin A (.alpha.2.beta.2), and it has been established
that these disorders can be ameliorated by reactivating production
of fetal hemoglobin (HbF, .alpha.2.gamma.2) in the patients' blood.
Even small increments in fetal hemoglobin decreases morbidity and
mortality in sickle cell disease, while higher levels are necessary
to completely ameliorate the symptoms. In .beta.-thalassemia,
increases in fetal globin synthesis, which reduces the excess
unbalanced .alpha.-globin chains by 10%, is often enough to
decrease the anemia to a level which does not require regular blood
transfusions.
[0009] Short chain fatty acids and derivatives of 2-9 carbons
induce expression of .gamma.-globin in cultured erythroid cells,
animal models and reporter gene assays, which test activity in
activating the .gamma.-globin gene promoter. Several short chain
fatty acids induce the .gamma.-globin promoter and have biologic
and clinical activity. Pharmacological re-introduction of HbF has
been achieved in patients with a prototype short-chain fatty acid,
arginine butyrate, resulting in sufficient levels of HbF to
ameliorate anemia and reduce clinical complications. Patients
treated in a Phase II trial with pulsed butyrate have experienced
both biochemical and clinical improvement in their diseases, with
excellent safety profiles. However, the prototype short chain fatty
acids have limitations as therapeutics. Arginine butyrate and
phenylbutyrate require 100 .mu.M levels in vitro and are rapidly
metabolized in vivo, necessitating large quantities (20 g for
sodium phenyl butyrate), an intravenous infusion for arginine
butyrate and careful adjustment of dosing to prevent secondary
suppression of erythopoiesis.
[0010] While advances have been made in this field, there remains a
need for new and/or improved methods for treating and preventing
blood disorders generally as well as for compounds and
pharmaceutical compositions for the same.
BRIEF SUMMARY
[0011] The present invention pertains, at least in part, to methods
for treating or preventing a blood disorder in a subject by
administering to the subject an effective amount of a compound of
formula I: ##STR1## wherein
[0012] R.sup.1 is hydroxy or alkoxy;
[0013] X is C(O), C(S), SO, SO.sub.2 or PO.sub.2;
[0014] R.sup.2 and R.sup.3 are each independently hydrogen, alkyl,
halogen, hydroxyl, thiol, alkenyl, alkynyl, aryl, acyl, alkoxy,
amino, alkylamino or heterocyclic;
[0015] R.sup.4 is alkyl, cycloalkyl, alkenyl, alkynyl, arylalkyl,
amido, alkylamino, amino, arylamino, alkylcarbonyl, arylcarbonyl,
alkylaminocarbonyl, alkoxy, alkoxycarbonyl, alkylcarbonyloxy,
alkyloxycarbonyloxy, arylcarbonyloxy, aryloxy, thiol, alkylthio,
arylthio, alkenyl, heterocyclic, heteroaryl, halogen or
##STR2##
[0016] R.sup.5 is hydrogen, alkyl, alkenyl, alkynyl, aryl,
arylalkyl, amido, alkylamino, amino, arylamino, alkylcarbonyl,
arylcarbonyl, alkylaminocarbonyl, alkoxy, alkoxycarbonyl,
alkylcarbonyloxy, alkyloxycarbonyloxy, arylcarbonyloxy, aryloxy,
thiol, alkylthio, arylthio, alkenyl, heterocyclic, hydroxy, nitro,
halogen or optionally linked to R.sup.6 to form a ring;
[0017] R.sup.6 is hydrogen, alkyl, alkenyl, alkynyl, aryl,
arylalkyl, amido, alkylamino, amino, arylamino, alkylcarbonyl,
arylcarbonyl, alkylaminocarbonyl, alkoxy, alkoxycarbonyl,
alkylcarbonyloxy, alkyloxycarbonyloxy, arylcarbonyloxy, aryloxy,
thiol, alkylthio, arylthio, alkenyl, heterocyclic, hydroxy, nitro,
halogen or optionally linked to R.sup.5 or R.sup.7 to form a
ring;
[0018] R.sup.7 is hydrogen, alkyl, alkenyl, alkynyl, aryl,
arylalkyl, amido, alkylamino, amino, arylamino, alkylcarbonyl,
arylcarbonyl, alkylaminocarbonyl, alkoxy, alkoxycarbonyl,
alkylcarbonyloxy, alkyloxycarbonyloxy, arylcarbonyloxy, aryloxy,
thiol, alkylthio, arylthio, alkenyl, heterocyclic, hydroxy, nitro,
halogen or optionally linked to R.sup.6 or R.sup.8 to form a
ring;
[0019] R.sup.8 is hydrogen, alkyl, alkenyl, alkynyl, aryl,
arylalkyl, amido, alkylamino, amino, arylamino, alkylcarbonyl,
arylcarbonyl, alkylaminocarbonyl, alkoxy, alkoxycarbonyl,
alkylcarbonyloxy, alkyloxycarbonyloxy, arylcarbonyloxy, aryloxy,
thiol, alkylthio, arylthio, alkenyl, heterocyclic, hydroxy, nitro,
halogen or optionally linked to R.sup.7 or R.sup.9 to form a
ring;
[0020] R.sup.9 is hydrogen, alkyl, alkenyl, alkynyl, aryl,
arylalkyl, amido, alkylamino, amino, arylamino, alkylcarbonyl,
arylcarbonyl, alkylaminocarbonyl, alkoxy, alkoxycarbonyl,
alkylcarbonyloxy, alkyloxycarbonyloxy, arylcarbonyloxy, aryloxy,
thiol, alkylthio, arylthio, alkenyl, heterocyclic, hydroxy, nitro,
halogen or optionally linked to R.sup.8 to form a ring;
[0021] and racemates, isolated enantiomers or diastereomers, and
pharmaceutically acceptable salts thereof;
[0022] provided that when R.sup.4 is ##STR3## R.sup.5, R.sup.6,
R.sup.7, R.sup.8, and R.sup.9 are each not hydrogen; and when
R.sup.6, R.sup.7, R.sup.8, and R.sup.9 are each hydrogen, R.sup.5
is not methoxy; and when R.sup.5, R.sup.7, R.sup.8, R.sup.9 are
hydrogen, R.sup.6 is not methoxy; and when R.sup.5, R.sup.8 and
R.sup.9 are hydrogen, R.sup.6 and R.sup.7 are not methoxy.
[0023] In another embodiment, the present invention pertains, at
least in part, to methods for treating or preventing a blood
disorder in a subject by administering to the subject an effective
amount of a compound of formula II: ##STR4## wherein
[0024] R.sup.1' is hydroxy or alkoxy;
[0025] Y is C(O);
[0026] n is 0 or an integer from 1 to 5;
[0027] R.sup.10 and R.sup.10' are each independently hydrogen,
alkyl, halogen, hydroxyl, thiol, alkenyl, alkynyl, aryl, acyl,
alkoxy, amino, alkylamino, heterocyclic or optionally joined to
form a ring;
[0028] R.sup.11 is CR.sup.11'R.sup.11''R.sup.11''', alkenyl,
cycloalkyl, alkynyl, arylalkyl, amido, alkylamino, amino,
arylamino, carbonylalkyl, alkylcarbonyl, arylcarbonyl,
alkylaminocarbonyl, alkoxycarbonyl, alkylcarbonyloxy,
alkyloxycarbonyloxy, arylcarbonyloxy, thiol, alkylthio, arylthio,
alkenyl, heterocyclic, heteroaryl, hydroxy, halogen or ##STR5##
[0029] R.sup.11' and R.sup.11'' are each independently hydrogen,
alkyl, alkenyl, alkynyl, aryl, arylalkyl, amido, alkylamino, amino,
arylamino, alkylcarbonyl, arylcarbonyl, alkylaminocarbonyl, alkoxy,
alkoxycarbonyl, alkylcarbonyloxy, alkyloxycarbonyloxy,
arylcarbonyloxy, aryloxy, thiol, alkylthio, arylthio, alkenyl,
heterocyclic, hydroxyl, halogen, or R.sup.11' and R.sup.11'' are
optionally joined to form a ring;
[0030] R.sup.11''' is alkenyl, alkynyl, aryl, arylalkyl, amido,
alkylamino, amino, arylamino, alkylcarbonyl, arylcarbonyl,
alkylaminocarbonyl, alkoxy, alkoxycarbonyl, alkylcarbonyloxy,
alkyloxycarbonyloxy, arylcarbonyloxy, aryloxy, thiol, alkylthio,
arylthio, alkenyl, heterocyclic, hydroxy or halogen;
[0031] R.sup.12 is hydrogen, alkenyl, alkynyl, aryl, arylalkyl,
amido, alkylamino, amino, arylamino, alkylcarbonyl, arylcarbonyl,
alkylaminocarbonyl, alkoxy, alkoxycarbonyl, alkylcarbonyloxy,
alkyloxycarbonyloxy, arylcarbonyloxy, aryloxy, thiol, alkylthio,
arylthio, alkenyl, heterocyclic or optionally linked to R.sup.13 to
form a ring;
[0032] R.sup.13 is hydrogen, alkyl, alkenyl, alkynyl, aryl,
arylalkyl, amido, alkylamino, amino, arylamino, alkylcarbonyl,
arylcarbonyl, alkylaminocarbonyl, alkoxy, alkoxycarbonyl,
alkylcarbonyloxy, alkyloxycarbonyloxy, arylcarbonyloxy, aryloxy,
thiol, alkylthio, arylthio, alkenyl, heterocyclic, hydroxy or
optionally linked to R.sup.12 or R.sup.14 to form a ring;
[0033] R.sup.14 is hydrogen, alkenyl, alkynyl, aryl, arylalkyl,
amido, alkylamino, amino, arylamino, alkylcarbonyl, arylcarbonyl,
alkylaminocarbonyl, alkoxy, alkoxycarbonyl, alkylcarbonyloxy,
alkyloxycarbonyloxy, arylcarbonyloxy, aryloxy, thiol, alkylthio,
arylthio, alkenyl, heterocyclic, halogen or optionally linked to
R.sup.13 or R.sup.15 to form a ring;
[0034] R.sup.15 is hydrogen, alkenyl, alkynyl, aryl, arylalkyl,
amido, alkylamino, amino, arylamino, alkylcarbonyl, arylcarbonyl,
alkylaminocarbonyl, alkoxy, alkoxycarbonyl, alkylcarbonyloxy,
alkyloxycarbonyloxy, arylcarbonyloxy, aryloxy, thiol, alkylthio,
arylthio, alkenyl, heterocyclic, halogen or optionally linked to
R.sup.14 or R.sup.16 to form a ring;
[0035] R.sup.16 is hydrogen, alkenyl, alkynyl, aryl, arylalkyl,
amido, alkylamino, amino, arylamino, alkylcarbonyl, arylcarbonyl,
alkylaminocarbonyl, alkoxy, alkoxycarbonyl, alkylcarbonyloxy,
alkyloxycarbonyloxy, arylcarbonyloxy, aryloxy, thiol, alkylthio,
arylthio, alkenyl, heterocyclic, halogen or optionally linked to
R.sup.15 to form a ring;
[0036] and racemates, isolated enantiomers or diastereomers, and
pharmaceutically acceptable salts thereof;
[0037] provided that when R.sup.11 is ##STR6## R.sup.12, R.sup.13,
R.sup.14, R.sup.15, and R.sup.16 are each not hydrogen; and
provided when n is 2, R.sup.11 is ##STR7## and R.sup.10, R.sup.10',
R.sup.12, R.sup.15, and R.sup.16 are hydrogen, then R.sup.14 and
R.sup.15 are not methoxy; and provided when n is 1, R.sup.11 is
##STR8## and R.sup.10, R.sup.10', R.sup.13, R.sup.14, and R.sup.16
are hydrogen, then R.sup.12 and R.sup.15 are not methoxy.
[0038] In yet another embodiment, the present invention pertains,
at least in part, to methods for treating or preventing a blood
disorder in a subject by administering to the subject an effective
amount of a compound of formula III: ##STR9## wherein
[0039] R.sup.1'' is hydroxy or alkoxy;
[0040] Z is C(S), SO, SO.sub.2 or PO.sub.2;
[0041] m is 0 or an integer from 1-5;
[0042] R.sup.17 and R.sup.17' are each independently hydrogen,
alkyl, halogen, hydroxyl, thiol, alkenyl, alkynyl, aryl, acyl,
alkoxy, amino, alkylamino or heterocyclic;
[0043] R.sup.18 is hydrogen, alkyl, cycloalkyl, alkenyl, alkynyl,
aryl, arylalkyl, amido, alkylamino, amino, arylamino,
alkylcarbonyl, arylcarbonyl, alkylaminocarbonyl, alkoxy,
alkoxycarbonyl, alkylcarbonyloxy, alkyloxycarbonyloxy,
arylcarbonyloxy, aryloxy, thiol, alkylthio, arylthio, alkenyl,
heterocyclic, heteroaryl, hydroxyl or halogen;
[0044] and racemates, isolated enantiomers or diastereomers, and
pharmaceutically acceptable salts thereof.
[0045] In another embodiment, the present invention pertains, at
least in part, to methods for treating or preventing a blood
disorder in a subject by administering to the subject an effective
amount of a compound of formula: ##STR10## In one embodiment, the
present invention pertains, at least in part, to pharmaceutical
compositions of an effective amount of a compound of formula I,
formula II, formula III, or ##STR11## and a pharmaceutically
acceptable carrier.
[0046] In another embodiment, the present invention pertains, at
least in part, to compounds of formula I, formula II, and formula
(III) and racemates, isolated enantiomers or diastereomers, and
pharmaceutically acceptable salts thereof.
DETAILED DESCRIPTION
[0047] In one embodiment, the present invention pertains, at least
in part, to methods
[0048] for treating or preventing a blood disorder in a subject by
administering to the subject an effective amount of a compound of
formula I: ##STR12## wherein
[0049] R.sup.1 is hydroxy or alkoxy;
[0050] X is C(O), C(S), SO, SO.sub.2 or PO.sub.2;
[0051] R.sup.2 and R.sup.3 are each independently hydrogen, alkyl,
halogen, hydroxyl, thiol, alkenyl, alkynyl, aryl, acyl, alkoxy,
amino, alkylamino or heterocyclic;
[0052] R.sup.4 is alkyl, cycloalkyl, alkenyl, alkynyl, arylalkyl,
amido, alkylamino, amino, arylamino, alkylcarbonyl, arylcarbonyl,
alkylaminocarbonyl, alkoxy, alkoxycarbonyl, alkylcarbonyloxy,
alkyloxycarbonyloxy, arylcarbonyloxy, aryloxy, thiol, alkylthio,
arylthio, alkenyl, heterocyclic, heteroaryl, halogen or
##STR13##
[0053] R.sup.5 is hydrogen, alkyl, alkenyl, alkynyl, aryl,
arylalkyl, amido, alkylamino, amino, arylamino, alkylcarbonyl,
arylcarbonyl, alkylaminocarbonyl, alkoxy, alkoxycarbonyl,
alkylcarbonyloxy, alkyloxycarbonyloxy, arylcarbonyloxy, aryloxy,
thiol, alkylthio, arylthio, alkenyl, heterocyclic, hydroxy, nitro,
halogen or optionally linked to R.sup.6 to form a ring;
[0054] R.sup.6 is hydrogen, alkyl, alkenyl, alkynyl, aryl,
arylalkyl, amido, alkylamino, amino, arylamino, alkylcarbonyl,
arylcarbonyl, alkylaminocarbonyl, alkoxy, alkoxycarbonyl,
alkylcarbonyloxy, alkyloxycarbonyloxy, arylcarbonyloxy, aryloxy,
thiol, alkylthio, arylthio, alkenyl, heterocyclic, hydroxy, nitro,
halogen or optionally linked to R.sup.5 or R.sup.7 to form a
ring;
[0055] R.sup.7 is hydrogen, alkyl, alkenyl, alkynyl, aryl,
arylalkyl, amido, alkylamino, amino, arylamino, alkylcarbonyl,
arylcarbonyl, alkylaminocarbonyl, alkoxy, alkoxycarbonyl,
alkylcarbonyloxy, alkyloxycarbonyloxy, arylcarbonyloxy, aryloxy,
thiol, alkylthio, arylthio, alkenyl, heterocyclic, hydroxy, nitro,
halogen or optionally linked to R.sup.6 or R.sup.8 to form a
ring;
[0056] R.sup.8 is hydrogen, alkyl, alkenyl, alkynyl, aryl,
arylalkyl, amido, alkylamino, amino, arylamino, alkylcarbonyl,
arylcarbonyl, alkylaminocarbonyl, alkoxy, alkoxycarbonyl,
alkylcarbonyloxy, alkyloxycarbonyloxy, arylcarbonyloxy, aryloxy,
thiol, alkylthio, arylthio, alkenyl, heterocyclic, hydroxy, nitro,
halogen or optionally linked to R.sup.7 or R.sup.9 to form a
ring;
[0057] R.sup.9 is hydrogen, alkyl, alkenyl, alkynyl, aryl,
arylalkyl, amido, alkylamino, amino, arylamino, alkylcarbonyl,
arylcarbonyl, alkylaminocarbonyl, alkoxy, alkoxycarbonyl,
alkylcarbonyloxy, alkyloxycarbonyloxy, arylcarbonyloxy, aryloxy,
thiol, alkylthio, arylthio, alkenyl, heterocyclic, hydroxy, nitro,
halogen or optionally linked to R.sup.9 to form a ring;
[0058] and racemates, isolated enantiomers or diastereomers, and
pharmaceutically acceptable salts thereof;
[0059] provided that when R.sup.4 is ##STR14## R.sup.5, R.sup.6,
R.sup.7, R.sup.8, and R.sup.9 are each not hydrogen; and when
R.sup.6, R.sup.7, R.sup.8, and R.sup.9 are each hydrogen, R.sup.5
is not methoxy; and when R.sup.5, R.sup.7, R.sup.8, R.sup.9 are
hydrogen, R.sup.6 is not methoxy; and when R.sup.5, R.sup.8 and
R.sup.9 are hydrogen, R.sup.6 and R.sup.7 are not methoxy.
[0060] In one embodiment, R.sup.1 is hydroxy, X is C(O) and R.sup.4
is ##STR15##
[0061] In one embodiment, R.sup.2, R.sup.3, R.sup.5, R.sup.6,
R.sup.8 and R.sup.9 are each hydrogen and R.sup.7 is alkoxy (e.g.,
methoxy).
[0062] In another embodiment, R.sup.2, R.sup.3, R.sup.5, R.sup.7
and R.sup.8 are each hydrogen, and R.sup.6 and R.sup.9 are each
alkyl (e.g., methyl).
[0063] In yet an other embodiment, R.sup.2, R.sup.3, R.sup.5,
R.sup.8 are R.sup.9 are each hydrogen and R.sup.6 and R.sup.7 are
each hydroxyl.
[0064] In a further embodiment, R.sup.2, R.sup.3, R.sup.5, R.sup.8
are R.sup.9 are each hydrogen and R.sup.6 and R.sup.7 are linked by
--O--CH.sub.2--O-- to form a ring.
[0065] In yet another embodiment, R.sup.2, R.sup.3, R.sup.5,
R.sup.6 and R.sup.9 are each hydrogen, R.sup.7 is alkoxy (e.g.,
methoxy) and R.sup.8 is hydroxyl.
[0066] In another embodiment, R.sup.1 is hydroxy, X is C(O) and
R.sup.4 is heteroaryl, such quinoline or substituted or
unsubstituted thiophene (e.g., chlorothiophene).
[0067] In one embodiment, the present invention pertains, at least
in part, to methods
[0068] for treating or preventing a blood disorder in a subject by
administering to the subject an effective amount of a compound of
formula I, wherein the compound of formula I is: ##STR16##
[0069] In another embodiment, the present invention pertains, at
least in part, to methods for treating or preventing a blood
disorder in a subject by administering to the subject an effective
amount of a compound of formula II: ##STR17## wherein
[0070] R.sup.1' is hydroxy or alkoxy;
[0071] Y is C(O);
[0072] n is 0 or an integer from 1 to 5;
[0073] R.sup.10 and R.sup.10' are each independently hydrogen,
alkyl, halogen, hydroxyl, thiol, alkenyl, alkynyl, aryl, acyl,
alkoxy, amino, alkylamino, heterocyclic or optionally joined to
form a ring;
[0074] R.sup.11 is CR.sup.11'R.sup.11''R.sup.11''', alkenyl,
cycloalkyl, alkynyl, arylalkyl, amido, alkylamino, amino,
arylamino, carbonylalkyl, alkylcarbonyl, arylcarbonyl,
alkylaminocarbonyl, alkoxycarbonyl, alkylcarbonyloxy,
alkyloxycarbonyloxy, arylcarbonyloxy, thiol, alkylthio, arylthio,
alkenyl, heterocyclic, heteroaryl, hydroxy, halogen or
##STR18##
[0075] R.sup.11' and R.sup.11'' are each independently hydrogen,
alkyl, alkenyl, alkynyl, aryl, arylalkyl, amido, alkylamino, amino,
arylamino, alkylcarbonyl, arylcarbonyl, alkylaminocarbonyl, alkoxy,
alkoxycarbonyl, alkylcarbonyloxy, alkyloxycarbonyloxy,
arylcarbonyloxy, aryloxy, thiol, alkylthio, arylthio, alkenyl,
heterocyclic, hydroxyl, halogen or R.sup.11' and R.sup.11'' are
optionally joined to form a ring;
[0076] R.sup.11''' is alkenyl, alkynyl, aryl, arylalkyl, amido,
alkylamino, amino, arylamino, alkylcarbonyl, arylcarbonyl,
alkylaminocarbonyl, alkoxy, alkoxycarbonyl, alkylcarbonyloxy,
alkyloxycarbonyloxy, arylcarbonyloxy, aryloxy, thiol, alkylthio,
arylthio, alkenyl, heterocyclic, hydroxy or halogen;
[0077] R.sup.12 is hydrogen, alkenyl, alkynyl, aryl, arylalkyl,
amido, alkylamino, amino, arylamino, alkylcarbonyl, arylcarbonyl,
alkylaminocarbonyl, alkoxy, alkoxycarbonyl, alkylcarbonyloxy,
alkyloxycarbonyloxy, arylcarbonyloxy, aryloxy, thiol, alkylthio,
arylthio, alkenyl, heterocyclic or optionally linked to R.sup.13 to
form a ring;
[0078] R.sup.13 is hydrogen, alkyl, alkenyl, alkynyl, aryl,
arylalkyl, amido, alkylamino, amino, arylamino, alkylcarbonyl,
arylcarbonyl, alkylaminocarbonyl, alkoxy, alkoxycarbonyl,
alkylcarbonyloxy, alkyloxycarbonyloxy, arylcarbonyloxy, aryloxy,
thiol, alkylthio, arylthio, alkenyl, heterocyclic, hydroxy or
optionally linked to R.sup.12 or R.sup.14 to form a ring;
[0079] R.sup.14 is hydrogen, alkenyl, alkynyl, aryl, arylalkyl,
amido, alkylamino, amino, arylamino, alkylcarbonyl, arylcarbonyl,
alkylaminocarbonyl, alkoxy, alkoxycarbonyl, alkylcarbonyloxy,
alkyloxycarbonyloxy, arylcarbonyloxy, aryloxy, thiol, alkylthio,
arylthio, alkenyl, heterocyclic, halogen or optionally linked to
R.sup.13 or R.sup.15 to form a ring;
[0080] R.sup.15 is hydrogen, alkenyl, alkynyl, aryl, arylalkyl,
amido, alkylamino, amino, arylamino, alkylcarbonyl, arylcarbonyl,
alkylaminocarbonyl, alkoxy, alkoxycarbonyl, alkylcarbonyloxy,
alkyloxycarbonyloxy, arylcarbonyloxy, aryloxy, thiol, alkylthio,
arylthio, alkenyl, heterocyclic, halogen or optionally linked to
R.sup.14 or R.sup.16 to form a ring;
[0081] R.sup.16 is hydrogen, alkenyl, alkynyl, aryl, arylalkyl,
amido, alkylamino, amino, arylamino, alkylcarbonyl, arylcarbonyl,
alkylaminocarbonyl, alkoxy, alkoxycarbonyl, alkylcarbonyloxy,
alkyloxycarbonyloxy, arylcarbonyloxy, aryloxy, thiol, alkylthio,
arylthio, alkenyl, heterocyclic, halogen or optionally linked to
R.sup.15 to form a ring;
[0082] and racemates, isolated enantiomers or diastereomers, and
pharmaceutically acceptable salts thereof;
[0083] provided that when R.sup.11 is ##STR19## R.sup.12, R.sup.13,
R.sup.14, R.sup.15, and R.sup.16 are each not hydrogen; and
provided when n is 2, R.sup.11 is ##STR20## and R.sup.10,
R.sup.10', R.sup.12, R.sup.15, and R.sup.16 are hydrogen, then
R.sup.14 and R.sup.15 are not methoxy; and provided when n is 1,
R.sup.11 is ##STR21## and R.sup.10, R.sup.10', R.sup.13, R.sup.14,
and R.sup.16 are hydrogen, then R.sup.12 and R.sup.15 are not
methoxy.
[0084] In one embodiment, R.sup.1' is hydroxyl, n is 5, R.sup.10
and R.sup.10' are hydrogen and R.sup.11 is alkylcarbonyl.
[0085] In another embodiment, R.sup.1' is hydroxyl, n is 2,
R.sup.10 and R.sup.10' are each hydrogen and R.sup.11 is
CR.sup.11'R.sup.11''R.sup.11'''.
[0086] In a further embodiment, R.sup.11' and R.sup.11'' are joined
by --(CH.sub.2).sub.5-- to form a cyclohexyl ring and R.sup.11'''
is a substituted or unsubstituted heterocycle (e.g.,
chlorothiophene).
[0087] In one embodiment, R.sup.1' is hydroxyl, n is 0 and R.sup.11
is ##STR22## In one embodiment, R.sup.13, R.sup.14, R.sup.15, and
R.sup.16 are hydrogen and R.sup.12 is arylthioalkyl or alkoxy
substituted aryloxy.
[0088] In another embodiment, R.sup.12, R.sup.14, R.sup.15 and
R.sup.16 are hydrogen and R.sup.13 is a substituted or
unsubstituted heterocycle, such as, for example, chromen-2-one,
nitro-substituted pyrazole, or chloro-substituted pyrazole.
[0089] In yet another embodiment, R.sup.12, R.sup.13, R.sup.15 and
R.sup.16 are hydrogen and R.sup.13 is alkoxy (e.g., ethoxy).
[0090] In a further embodiment, R.sup.12, R.sup.15 and R.sup.16 are
hydrogen, and R.sup.13 and R.sup.14 are linked by
--N(H)C(O)CH.sub.2S-- to form a ring.
[0091] In yet another embodiment, R.sup.14, R.sup.15 and R.sup.16
are each hydrogen, and R.sup.12 and R.sup.13 are linked by
--CH.dbd.C(CH.sub.3)O-- to form a ring.
[0092] In one embodiment, R.sup.1' is hydroxyl, n is 0 and R.sup.11
is a substituted or unsubstituted heterocycle (e.g., substituted
aryl-substituted furan) or a substituted or unsubstituted
cycloalkyl (e.g., tetrahydrobenzothiadiazole or
dihydrobenzothiophenone).
[0093] In one embodiment, R.sup.1' is hydroxyl, n is 1, R.sup.10
and R.sup.10' are hydrogen and R.sup.11 is a substituted or
unsubstituted cycloalkyl (e.g., dimethylcyclobutane carboxylic
acid).
[0094] In another embodiment, R.sup.1' is hydroxyl, n is 1,
R.sup.10 and R.sup.10' are hydrogen and R.sup.11 is ##STR23##
[0095] In one embodiment, R.sup.12, R.sup.13 and R.sup.16 are
hydrogen, and R.sup.14 and R.sup.15 are linked by
--O--CH.sub.2--O-- to form a ring.
[0096] In another embodiment, R.sup.1' is hydroxyl, n is 1,
R.sup.10 and R.sup.10' are hydrogen and R.sup.11 is a substituted
or unsubstituted heterocycle, such as, for example substituted
thiazolidinedione, substituted pyridinone or substituted
pyrazole.
[0097] In yet another embodiment, R.sup.1' is hydroxyl, n is 1,
R.sup.10 and R.sup.10' are hydrogen and R.sup.11 is a unsubstituted
or substituted arylamino (e.g., trifluorothio-substituted
arylamino).
[0098] In a further embodiment, R.sup.1' is hydroxyl, n is 1,
R.sup.10 and R.sup.10' are hydrogen and R.sup.11 is unsubstituted
or substituted arylthio (e.g., methoxyphenylthio) or unsubstituted
or substituted heterocyclic thio, such as, for example, substituted
triazolethio, substituted thiadiazolethio or substituted
thiophenethio.
[0099] In another embodiment, R.sup.1' is hydroxyl, n is 1,
R.sup.10 and R.sup.10' are hydrogen and R.sup.11 is R.sup.11 is
CR.sup.11'R.sup.11''R.sup.11''' and R.sup.11' is hydrogen,
R.sup.11'' is amino and R.sup.11''' is alkoxy-substituted aryl.
[0100] In one embodiment, R.sup.1' is hydroxyl, n is 1, R.sup.10 is
hydrogen and R.sup.10' is alkyl. In one embodiment, R.sup.10' is
isopropyl and R.sup.11 is substituted or unsubstituted arylthio,
such as, for example, alkoxy-substituted phenylthio or
alkoxy-substituted pyrimadinylthio.
[0101] In another embodiment, R.sup.10' is ethyl and R.sup.11 is
heteroarylamino (e.g., quinazolinylamino).
[0102] In yet another embodiment, R.sup.1' is hydroxyl, n is 1,
R.sup.10 and R.sup.10' are linked by --(CH.sub.2).sub.5-- to form a
cyclohexyl ring and R.sup.11 is heterocyclic substituted
carbonylalkyl.
[0103] In one embodiment, the compounds of formula (II) do not
include compounds wherein R.sup.1 is hydroxy, R.sup.10 is alkyl,
e.g., ethyl, R.sup.10' is hydrogen, n is 1, and R.sup.11 is
arylamino, e.g., quinazolin-4-ylamino.
[0104] In one embodiment, the compounds of formula (II) do not
include compounds wherein R.sup.1 is hydroxy, R.sup.10 is hydrogen,
R.sup.10' is alkyl, e.g., ethyl, n is 1, and R.sup.11 is arylamino,
e.g., quinazolin-4-ylamino.
[0105] In another embodiment, the compounds of formula (II) do not
include compounds wherein R.sup.1 is hydroxy, R.sup.10 and
R.sup.10' are each hydrogen, n is 1, and R.sup.11 is aryl amino
(e.g., trifluorothio-substituted arylamino).
[0106] In another embodiment, the compounds of formula (II) do not
include 2-(quinazolin-4-ylamino)butyric acid or
[4-[(trifluoromethyl)sulfanyl]-anilino]-acetic acid.
[0107] In one embodiment, the present invention pertains, at least
in part, to methods
[0108] for treating or preventing a blood disorder in a subject by
administering to the subject an effective amount of a compound of
formula II, wherein the compound of formula II is: ##STR24##
##STR25## ##STR26##
[0109] as well as racemates and isolated enantiomers and
diastereomers thereof.
[0110] In another embodiment, the present invention pertains, at
least in part, to methods for treating or preventing a blood
disorder in a subject by administering to the subject an effective
amount of a compound of formula III: ##STR27## wherein
[0111] R.sup.1'' is hydroxy or alkoxy;
[0112] Z is C(S), SO, SO.sub.2 or PO.sub.2;
[0113] m is 0 or an integer from 1-5;
[0114] R.sup.17 and R.sup.17' are each independently hydrogen,
alkyl, halogen, hydroxyl, thiol, alkenyl, alkynyl, aryl, acyl,
alkoxy, amino, alkylamino or heterocyclic;
[0115] R.sup.18 is hydrogen, alkyl, cycloalkyl, alkenyl, alkynyl,
aryl, arylalkyl, amido, alkylamino, amino, arylamino,
alkylcarbonyl, arylcarbonyl, alkylaminocarbonyl, alkoxy,
alkoxycarbonyl, alkylcarbonyloxy, alkyloxycarbonyloxy,
arylcarbonyloxy, aryloxy, thiol, alkylthio, arylthio, alkenyl,
heterocyclic, heteroaryl, hydroxyl or halogen;
[0116] and racemates, isolated enantiomers or diastereomers, and
pharmaceutically acceptable salts thereof.
[0117] In one embodiment, R.sup.1'' is hydroxyl, Z is SO.sub.2, m
is 0, and R.sup.18 is disubstituted aryl substituted by, for
example, nitro and fluoro.
[0118] In yet another embodiment, the present invention pertains,
at least in part, to methods for treating or preventing a blood
disorder in a subject by administering to the subject an effective
amount of a compound of formula III, wherein the compound of
formula III is: ##STR28##
[0119] In another embodiment, the present invention pertains, at
least in part, to methods for treating or preventing a blood
disorder in a subject by administering to the subject an effective
amount of a compound of formula: ##STR29##
[0120] In yet another embodiment, the invention pertains, at least
in part, to a compound of formula I: ##STR30## wherein
[0121] R.sup.1 is hydroxy or alkoxy;
[0122] X is C(O), C(S), SO, SO.sub.2 or PO.sub.2;
[0123] R.sup.2 and R.sup.3 are each independently hydrogen, alkyl,
halogen, hydroxyl, thiol, alkenyl, alkynyl, aryl, acyl, alkoxy,
amino, alkylamino or heterocyclic;
[0124] R.sup.4 is alkyl, cycloalkyl, alkenyl, alkynyl, arylalkyl,
amido, alkylamino, amino, arylamino, alkylcarbonyl, arylcarbonyl,
alkylaminocarbonyl, alkoxy, alkoxycarbonyl, alkylcarbonyloxy,
alkyloxycarbonyloxy, arylcarbonyloxy, aryloxy, thiol, alkylthio,
arylthio, alkenyl, heterocyclic, heteroaryl, halogen or
##STR31##
[0125] R.sup.5 is hydrogen, alkyl, alkenyl, alkynyl, aryl,
arylalkyl, amido, alkylamino, amino, arylamino, alkylcarbonyl,
arylcarbonyl, alkylaminocarbonyl, alkoxy, alkoxycarbonyl,
alkylcarbonyloxy, alkyloxycarbonyloxy, arylcarbonyloxy, aryloxy,
thiol, alkylthio, arylthio, alkenyl, heterocyclic, hydroxy, nitro,
halogen or optionally linked to R.sup.6 to form a ring;
[0126] R.sup.6 is hydrogen, alkyl, alkenyl, alkynyl, aryl,
arylalkyl, amido, alkylamino, amino, arylamino, alkylcarbonyl,
arylcarbonyl, alkylaminocarbonyl, alkoxy, alkoxycarbonyl,
alkylcarbonyloxy, alkyloxycarbonyloxy, arylcarbonyloxy, aryloxy,
thiol, alkylthio, arylthio, alkenyl, heterocyclic, hydroxy, nitro,
halogen or optionally linked to R.sup.5 or R.sup.7 to form a
ring;
[0127] R.sup.7 is hydrogen, alkyl, alkenyl, alkynyl, aryl,
arylalkyl, amido, alkylamino, amino, arylamino, alkylcarbonyl,
arylcarbonyl, alkylaminocarbonyl, alkoxy, alkoxycarbonyl,
alkylcarbonyloxy, alkyloxycarbonyloxy, arylcarbonyloxy, aryloxy,
thiol, alkylthio, arylthio, alkenyl, heterocyclic, hydroxy, nitro,
halogen or optionally linked to R.sup.6 or R.sup.8 to form a
ring;
[0128] R.sup.8 is hydrogen, alkyl, alkenyl, alkynyl, aryl,
arylalkyl, amido, alkylamino, amino, arylamino, alkylcarbonyl,
arylcarbonyl, alkylaminocarbonyl, alkoxy, alkoxycarbonyl,
alkylcarbonyloxy, alkyloxycarbonyloxy, arylcarbonyloxy, aryloxy,
thiol, alkylthio, arylthio, alkenyl, heterocyclic, hydroxy, nitro,
halogen or optionally linked to R.sup.7 or R.sup.9 to form a
ring;
[0129] R.sup.9 is hydrogen, alkyl, alkenyl, alkynyl, aryl,
arylalkyl, amido, alkylamino, amino, arylamino, alkylcarbonyl,
arylcarbonyl, alkylaminocarbonyl, alkoxy, alkoxycarbonyl,
alkylcarbonyloxy, alkyloxycarbonyloxy, arylcarbonyloxy, aryloxy,
thiol, alkylthio, arylthio, alkenyl, heterocyclic, hydroxy, nitro,
halogen or optionally linked to R.sup.8 to form a ring;
[0130] and racemates, isolated enantiomers or diastereomers, and
pharmaceutically acceptable salts thereof;
[0131] provided that when R.sup.4 is ##STR32## R.sup.5, R.sup.6,
R.sup.7, R.sup.8, and R.sup.9 are each not hydrogen; and when
R.sup.6, R.sup.7, R.sup.8, and R.sup.9 are each hydrogen, R.sup.5
is not methoxy; and when R.sup.5, R.sup.7, R.sup.8, R.sup.9 are
hydrogen, R.sup.6 is not methoxy; and when R.sup.5, R.sup.8 and
R.sup.9 are hydrogen, R.sup.6 and R.sup.7 are not methoxy;
[0132] provided that the compound is not a compound of the formula:
##STR33##
[0133] The present invention also pertains, at least in part, to a
compound of formula II: ##STR34## wherein
[0134] R.sup.1' is hydroxy or alkoxy;
[0135] Y is C(O);
[0136] n is 0 or an integer from 1 to 5;
[0137] R.sup.10 and R.sup.10' are each independently hydrogen,
alkyl, halogen, hydroxyl, thiol, alkenyl, alkynyl, aryl, acyl,
alkoxy, amino, alkylamino, heterocyclic or optionally joined to
form a ring;
[0138] R.sup.11 is CR.sup.11'R.sup.11''R.sup.11''', alkenyl,
cycloalkyl, alkynyl, arylalkyl, amido, alkylamino, amino,
arylamino, carbonylalkyl, alkylcarbonyl, arylcarbonyl,
alkylaminocarbonyl, alkoxycarbonyl, alkylcarbonyloxy,
alkyloxycarbonyloxy, arylcarbonyloxy, thiol, alkylthio, arylthio,
alkenyl, heterocyclic, heteroaryl, hydroxy, halogen or
##STR35##
[0139] R.sup.11' and R.sup.11'' are each independently hydrogen,
alkyl, alkenyl, alkynyl, aryl, arylalkyl, amido, alkylamino, amino,
arylamino, alkylcarbonyl, arylcarbonyl, alkylaminocarbonyl, alkoxy,
alkoxycarbonyl, alkylcarbonyloxy, alkyloxycarbonyloxy,
arylcarbonyloxy, aryloxy, thiol, alkylthio, arylthio, alkenyl,
heterocyclic, hydroxyl, halogen or R.sup.11' and R.sup.11'' are
optionally joined to form a ring;
[0140] R.sup.11''' is alkenyl, alkynyl, aryl, arylalkyl, amido,
alkylamino, amino, arylamino, alkylcarbonyl, arylcarbonyl,
alkylaminocarbonyl, alkoxy, alkoxycarbonyl, alkylcarbonyloxy,
alkyloxycarbonyloxy, arylcarbonyloxy, aryloxy, thiol, alkylthio,
arylthio, alkenyl, heterocyclic, hydroxy or halogen;
[0141] R.sup.12 is hydrogen, alkenyl, alkynyl, aryl, arylalkyl,
amido, alkylamino, amino, arylamino, alkylcarbonyl, arylcarbonyl,
alkylaminocarbonyl, alkoxy, alkoxycarbonyl, alkylcarbonyloxy,
alkyloxycarbonyloxy, arylcarbonyloxy, aryloxy, thiol, alkylthio,
arylthio, alkenyl, heterocyclic or optionally linked to R.sup.13 to
form a ring;
[0142] R.sup.13 is hydrogen, alkyl, alkenyl, alkynyl, aryl,
arylalkyl, amido, alkylamino, amino, arylamino, alkylcarbonyl,
arylcarbonyl, alkylaminocarbonyl, alkoxy, alkoxycarbonyl,
alkylcarbonyloxy, alkyloxycarbonyloxy, arylcarbonyloxy, aryloxy,
thiol, alkylthio, arylthio, alkenyl, heterocyclic, hydroxy or
optionally linked to R.sup.12 or R.sup.14 to form a ring;
[0143] R.sup.14 is hydrogen, alkenyl, alkynyl, aryl, arylalkyl,
amido, alkylamino, amino, arylamino, alkylcarbonyl, arylcarbonyl,
alkylaminocarbonyl, alkoxy, alkoxycarbonyl, alkylcarbonyloxy,
alkyloxycarbonyloxy, arylcarbonyloxy, aryloxy, thiol, alkylthio,
arylthio, alkenyl, heterocyclic, halogen or optionally linked to
R.sup.13 or R.sup.15 to form a ring;
[0144] R.sup.15 is hydrogen, alkenyl, alkynyl, aryl, arylalkyl,
amido, alkylamino, amino, arylamino, alkylcarbonyl, arylcarbonyl,
alkylaminocarbonyl, alkoxy, alkoxycarbonyl, alkylcarbonyloxy,
alkyloxycarbonyloxy, arylcarbonyloxy, aryloxy, thiol, alkylthio,
arylthio, alkenyl, heterocyclic, halogen or optionally linked to
R.sup.14 or R.sup.16 to form a ring;
[0145] R.sup.16 is hydrogen, alkenyl, alkynyl, aryl, arylalkyl,
amido, alkylamino, amino, arylamino, alkylcarbonyl, arylcarbonyl,
alkylaminocarbonyl, alkoxy, alkoxycarbonyl, alkylcarbonyloxy,
alkyloxycarbonyloxy, arylcarbonyloxy, aryloxy, thiol, alkylthio,
arylthio, alkenyl, heterocyclic, halogen or optionally linked to
R.sup.15 to form a ring;
[0146] and racemates, isolated enantiomers or diastereomers, and
pharmaceutically acceptable salts thereof;
[0147] provided that when R.sup.11 is ##STR36## R.sup.12, R.sup.13,
R.sup.14, R.sup.15, and R.sup.16 are each not hydrogen; and
provided when n is 2, R.sup.11 is ##STR37## and R.sup.10,
R.sup.10', R.sup.12, R.sup.15, and R.sup.16 are hydrogen, then
R.sup.14 and R.sup.15 are not methoxy; and provided when n is 1,
R.sup.11 is ##STR38## and R.sup.10, R.sup.10', R.sup.13, R.sup.14,
and R.sup.16 are hydrogen, then R.sup.12 and R.sup.15 are not
methoxy;
[0148] and provided that the compound is not a compound of:
##STR39## ##STR40##
[0149] In a further embodiment, the present invention pertains, at
least in part, to a compound of formula III: ##STR41## wherein
[0150] R.sup.1'' is hydroxy or alkoxy;
[0151] Z is C(S), SO, SO.sub.2 or PO.sub.2;
[0152] m is 0 or an integer from 1-5;
[0153] R.sup.17 and R.sup.17' are each independently hydrogen,
alkyl, halogen, hydroxyl, thiol, alkenyl, alkynyl, aryl, acyl,
alkoxy, amino, alkylamino or heterocyclic;
[0154] R.sup.18 is hydrogen, alkyl, cycloalkyl, alkenyl, alkynyl,
aryl, arylalkyl, amido, alkylamino, amino, arylamino,
alkylcarbonyl, arylcarbonyl, alkylaminocarbonyl, alkoxy,
alkoxycarbonyl, alkylcarbonyloxy, alkyloxycarbonyloxy,
arylcarbonyloxy, aryloxy, thiol, alkylthio, arylthio, alkenyl,
heterocyclic, heteroaryl, hydroxyl or halogen;
[0155] and racemates, isolated enantiomers or diastereomers, and
pharmaceutically acceptable salts thereof;
[0156] provided that the compound is not: ##STR42##
[0157] In one embodiment, the compounds of the invention do not
include the compounds described in S. Casteneda et al., Blood
Cells, Molecules, and Diseases, 35 (2005) 217-226.
[0158] The term "alkyl" includes saturated aliphatic groups,
including straight-chain alkyl groups (e.g., methyl, ethyl, propyl,
butyl, pentyl, hexyl, heptyl, octyl, nonyl, decyl, etc.),
branched-chain alkyl groups (isopropyl, tert-butyl, isobutyl,
etc.), cycloalkyl (alicyclic) groups (cyclopropyl, cyclopentyl,
cyclohexyl, cycloheptyl, cyclooctyl), alkyl substituted cycloalkyl
groups, and cycloalkyl substituted alkyl groups. The term alkyl
further includes alkyl groups, which can further include oxygen,
nitrogen, sulfur or phosphorous atoms replacing one or more carbons
of the hydrocarbon backbone. In certain embodiments, a straight
chain or branched chain alkyl has 6 or fewer carbon atoms in its
backbone (e.g., C.sub.1-C.sub.6 for straight chain, C.sub.3-C.sub.6
for branched chain), and more preferably 4 or fewer. Likewise,
preferred cycloalkyls have from 3-8 carbon atoms in their ring
structure, and more preferably have 5 or 6 carbons in the ring
structure. The term C.sub.1-C.sub.6 includes alkyl groups
containing 1 to 6 carbon atoms.
[0159] Moreover, the term alkyl includes both "unsubstituted
alkyls" and "substituted alkyls", the latter of which refers to
alkyl moieties having substituents replacing a hydrogen on one or
more carbons of the hydrocarbon backbone. Such substituents can
include, for example, alkenyl, alkynyl, halogen, hydroxyl,
alkylcarbonyloxy, arylcarbonyloxy, alkoxycarbonyloxy,
aryloxycarbonyloxy, carboxylate, alkylcarbonyl, arylcarbonyl,
alkoxycarbonyl, aminocarbonyl, alkylaminocarbonyl,
dialkylaminocarbonyl, alkylthiocarbonyl, alkoxyl, phosphate,
phosphonato, phosphinato, cyano, amino (including alkyl amino,
dialkylamino, arylamino, diarylamino, and alkylarylamino),
acylamino (including alkylcarbonylamino, arylcarbonylamino,
carbamoyl and ureido), amidino, imino, sulfhydryl, alkylthio,
arylthio, thiocarboxylate, sulfates, alkylsulfinyl, sulfonato,
sulfamoyl, sulfonamido, nitro, trifluoromethyl, cyano, azido,
heterocyclyl, alkylaryl, or an aromatic or heteroaromatic moiety.
Cycloalkyls can be further substituted, e.g., with the substituents
described above. An "alkylaryl" or an "arylalkyl" moiety is an
alkyl substituted with an aryl (e.g., phenylmethyl (benzyl)). The
term "alkyl" also includes the side chains of natural and unnatural
amino acids.
[0160] The term "aryl" includes groups, including 5- and 6-membered
single-ring aromatic groups that may include from zero to four
heteroatoms, for example, benzene, phenyl, pyrrole, furan,
thiophene, thiazole, isothiaozole, imidazole, triazole, tetrazole,
pyrazole, oxazole, isooxazole, pyridine, pyrazine, pyridazine, and
pyrimidine, and the like. Furthermore, the term "aryl" includes
multicyclic aryl groups, e.g., tricyclic, bicyclic, e.g.,
naphthalene, benzoxazole, benzodioxazole, benzothiazole,
benzoimidazole, benzothiophene, methylenedioxophenyl, quinoline,
isoquinoline, naphthridine, indole, benzofuran, purine, benzofuran,
deazapurine, or indolizine. Those aryl groups having heteroatoms in
the ring structure may also be referred to as "aryl heterocycles",
"heterocycles," "heteroaryls" or "heteroaromatics". The aromatic
ring can be substituted at one or more ring positions with such
substituents as described above, as for example, halogen, hydroxyl,
alkoxy, alkylcarbonyloxy, arylcarbonyloxy, alkoxycarbonyloxy,
aryloxycarbonyloxy, carboxylate, alkylcarbonyl,
alkylaminoacarbonyl, arylalkyl aminocarbonyl, alkenylaminocarbonyl,
alkylcarbonyl, arylcarbonyl, arylalkylcarbonyl, alkenylcarbonyl,
alkoxycarbonyl, aminocarbonyl, alkylthiocarbonyl, phosphate,
phosphonato, phosphinato, cyano, amino (including alkyl amino,
dialkylamino, arylamino, diarylamino, and alkylarylamino),
acylamino (including alkylcarbonylamino, arylcarbonylamino,
carbamoyl and ureido), amidino, imino, sulfhydryl, alkylthio,
arylthio, thiocarboxylate, sulfates, alkylsulfinyl, sulfonato,
sulfamoyl, sulfonamido, nitro, trifluoromethyl, cyano, azido,
heterocyclyl, alkylaryl, or an aromatic or heteroaromatic moiety,
Aryl groups can also be fused or bridged with alicyclic or
heterocyclic rings which are not aromatic so as to form a polycycle
(e.g., tetralin).
[0161] The term "alkenyl" includes unsaturated aliphatic groups
analogous in length and possible substitution to the alkyls
described above, but that contain at least one double bond.
[0162] For example, the term "alkenyl" includes straight-chain
alkenyl groups (e.g., ethylenyl, propenyl, butenyl, pentenyl,
hexenyl, heptenyl, octenyl, nonenyl, decenyl, etc.), branched-chain
alkenyl groups, cycloalkenyl (alicyclic) groups (cyclopropenyl,
cyclopentenyl, cyclohexenyl, cycloheptenyl, cyclooctenyl), alkyl or
alkenyl substituted cycloalkenyl groups, and cycloalkyl or
cycloalkenyl substituted alkenyl groups. The term alkenyl further
includes alkenyl groups which include oxygen, nitrogen, sulfur or
phosphorous atoms replacing one or more carbons of the hydrocarbon
backbone. In certain embodiments, a straight chain or branched
chain alkenyl group has 6 or fewer carbon atoms in its backbone
(e.g., C.sub.2-C.sub.6 for straight chain, C.sub.3-C.sub.6 for
branched chain). Likewise, cycloalkenyl groups may have from 3-8
carbon atoms in their ring structure, and more preferably have 5 or
6 carbons in the ring structure. The term C.sub.2-C.sub.6 includes
alkenyl groups containing 2 to 6 carbon atoms.
[0163] Moreover, the term alkenyl includes both "unsubstituted
alkenyls" and "substituted alkenyls", the latter of which refers to
alkenyl moieties having substituents replacing a hydrogen on one or
more carbons of the hydrocarbon backbone. Such substituents can
include, for example, alkyl groups, alkynyl groups, halogens,
hydroxyl, alkylcarbonyloxy, arylcarbonyloxy, alkoxycarbonyloxy,
aryloxycarbonyloxy, carboxylate, alkylcarbonyl, arylcarbonyl,
alkoxycarbonyl, aminocarbonyl, alkylaminocarbonyl,
dialkylaminocarbonyl, alkylthiocarbonyl, alkoxyl, phosphate,
phosphonato, phosphinato, cyano, amino (including alkyl amino,
dialkylamino, arylamino, diarylamino, and alkylarylamino),
acylamino (including alkylcarbonylamino, arylcarbonylamino,
carbamoyl and ureido), amidino, imino, sulfhydryl, alkylthio,
arylthio, thiocarboxylate, sulfates, alkylsulfinyl, sulfonato,
sulfamoyl, sulfonamido, nitro, trifluoromethyl, cyano, azido,
heterocyclyl, alkylaryl, or an aromatic or heteroaromatic
moiety.
[0164] The term "alkynyl" includes unsaturated aliphatic groups
analogous in length and possible substitution to the alkyls
described above, but which contain at least one triple bond.
[0165] For example, the term "alkynyl" includes straight-chain
alkynyl groups (e.g., ethynyl, propynyl, butynyl, pentynyl,
hexynyl, heptynyl, octynyl, nonynyl, decynyl, etc.), branched-chain
alkynyl groups, and cycloalkyl or cycloalkenyl substituted alkynyl
groups. The term alkynyl further includes alkynyl groups which
include oxygen, nitrogen, sulfur or phosphorous atoms replacing one
or more carbons of the hydrocarbon backbone. In certain
embodiments, a straight chain or branched chain alkynyl group has 6
or fewer carbon atoms in its backbone (e.g., C.sub.2-C.sub.6 for
straight chain, C.sub.3-C.sub.6 for branched chain). The term
C.sub.2-C.sub.6 includes alkynyl groups containing 2 to 6 carbon
atoms.
[0166] Moreover, the term alkynyl includes both "unsubstituted
alkynyls" and "substituted alkynyls", the latter of which refers to
alkynyl moieties having substituents replacing a hydrogen on one or
more carbons of the hydrocarbon backbone. Such substituents can
include, for example, alkyl groups, alkynyl groups, halogens,
hydroxyl, alkylcarbonyloxy, arylcarbonyloxy, alkoxycarbonyloxy,
aryloxycarbonyloxy, carboxylate, alkylcarbonyl, arylcarbonyl,
alkoxycarbonyl, aminocarbonyl, alkylaminocarbonyl,
dialkylaminocarbonyl, alkylthiocarbonyl, alkoxyl, phosphate,
phosphonato, phosphinato, cyano, amino (including alkyl amino,
dialkylamino, arylamino, diarylamino, and alkylarylamino),
acylamino (including alkylcarbonylamino, arylcarbonylamino,
carbamoyl and ureido), amidino, imino, sulfhydryl, alkylthio,
arylthio, thiocarboxylate, sulfates, alkylsulfinyl, sulfonato,
sulfamoyl, sulfonamido, nitro, trifluoromethyl, cyano, azido,
heterocyclyl, alkylaryl, or an aromatic or heteroaromatic
moiety.
[0167] Unless the number of carbons is otherwise specified, "lower
alkyl" as used herein means an alkyl group, as defined above, but
having from one to five carbon atoms in its backbone structure.
"Lower alkenyl" and "lower alkynyl" have chain lengths of, for
example, 2-5 carbon atoms.
[0168] The term "acyl" includes compounds and moieties which
contain the acyl radical (CH.sub.3CO--) or a carbonyl group. It
includes substituted acyl moieties. The term "substituted acyl"
includes acyl groups where one or more of the hydrogen atoms are
replaced by for example, alkyl groups, alkynyl groups, halogens,
hydroxyl, alkylcarbonyloxy, arylcarbonyloxy, alkoxycarbonyloxy,
aryloxycarbonyloxy, carboxylate, alkylcarbonyl, arylcarbonyl,
alkoxycarbonyl, aminocarbonyl, alkylaminocarbonyl,
dialkylaminocarbonyl, alkylthiocarbonyl, alkoxyl, phosphate,
phosphonato, phosphinato, cyano, amino (including alkyl amino,
dialkylamino, arylamino, diarylamino, and alkylarylamino),
acylamino (including alkylcarbonylamino, arylcarbonylamino,
carbamoyl and ureido), amidino, imino, sulfhydryl, alkylthio,
arylthio, thiocarboxylate, sulfates, alkylsulfinyl, sulfonato,
sulfamoyl, sulfonamido, nitro, trifluoromethyl, cyano, azido,
heterocyclyl, alkylaryl, or an aromatic or heteroaromatic
moiety.
[0169] The term "acylamino" includes moieties wherein an acyl
moiety is bonded to an amino group. For example, the term includes
alkylcarbonylamino, arylcarbonylamino, carbamoyl and ureido
groups.
[0170] The term "aroyl" includes compounds and moieties with an
aryl or heteroaromatic moiety bound to a carbonyl group. Examples
of aroyl groups include phenylcarboxy, naphthyl carboxy, etc.
[0171] The terms "alkoxyalkyl", "alkylaminoalkyl" and
"thioalkoxyalkyl" include alkyl groups, as described above, which
further include oxygen, nitrogen or sulfur atoms replacing one or
more carbons of the hydrocarbon backbone, e.g., oxygen, nitrogen or
sulfur atoms.
[0172] The term "alkoxy" includes substituted and unsubstituted
alkyl, alkenyl, and alkynyl groups covalently linked to an oxygen
atom. Examples of alkoxy groups include methoxy, ethoxy,
isopropyloxy, propoxy, butoxy, and pentoxy groups. Examples of
substituted alkoxy groups include halogenated alkoxy groups. The
alkoxy groups can be substituted with groups such as alkenyl,
alkynyl, halogen, hydroxyl, alkylcarbonyloxy, arylcarbonyloxy,
alkoxycarbonyloxy, aryloxycarbonyloxy, carboxylate, alkylcarbonyl,
arylcarbonyl, alkoxycarbonyl, aminocarbonyl, alkylaminocarbonyl,
dialkylaminocarbonyl, alkylthiocarbonyl, alkoxyl, phosphate,
phosphonato, phosphinato, cyano, amino (including alkyl amino,
dialkylamino, arylamino, diarylamino, and alkylarylamino),
acylamino (including alkylcarbonylamino, arylcarbonylamino,
carbamoyl and ureido), amidino, imino, sulfhydryl, alkylthio,
arylthio, thiocarboxylate, sulfates, alkylsulfinyl, sulfonato,
sulfamoyl, sulfonamido, nitro, trifluoromethyl, cyano, azido,
heterocyclyl, alkylaryl, or an aromatic or heteroaromatic moieties.
Examples of halogen substituted alkoxy groups include, but are not
limited to, fluoromethoxy, difluoromethoxy, trifluoromethoxy,
chloromethoxy, dichloromethoxy, trichloromethoxy, etc.
[0173] The term "amine" or "amino" includes compounds where a
nitrogen atom is covalently bonded to at least one carbon or
heteroatom. The term "alkyl amino" includes groups and compounds
wherein the nitrogen is bound to at least one additional alkyl
group. The term "dialkyl amino" includes groups wherein the
nitrogen atom is bound to at least two additional alkyl groups. The
term "arylamino" and "diarylamino" include groups wherein the
nitrogen is bound to at least one or two aryl groups, respectively.
The term "alkylarylamino," "alkylaminoaryl" or "arylaminoalkyl"
refers to an amino group which is bound to at least one alkyl group
and at least one aryl group. The term "alkaminoalkyl" or "alkyl
aminoalkyl" refers to an alkyl, alkenyl, or alkynyl group bound to
a nitrogen atom which is also bound to an alkyl group.
[0174] The term "amide" or "aminocarbonyl" includes compounds or
moieties which contain a nitrogen atom which is bound to the carbon
of a carbonyl or a thiocarbonyl group. The term includes
"alkaminocarbonyl" or "alkylaminocarbonyl" groups which include
alkyl, alkenyl, aryl or alkynyl groups bound to an amino group
bound to a carbonyl group. It includes arylaminocarbonyl groups
which include aryl or heteroaryl moieties bound to an amino group
which is bound to the carbon of a carbonyl or thiocarbonyl group.
The terms "alkylaminocarbonyl," "alkenylaminocarbonyl,"
"alkynylaminocarbonyl," arylaminocarbonyl," "alkylcarbonylamino,"
"alkenylcarbonylamino," alkynylcarbonylamino," and
"arylcarbonylamino" are included in term "amide." Amides also
include urea groups (aminocarbonylamino) and carbamates
(oxycarbonylamino).
[0175] The term "carbonyl" or "carboxy" includes compounds and
moieties which contain a carbon connected with a double bond to an
oxygen atom. The carbonyl can be further substituted with any
moiety which allows the compounds of the invention to perform its
intended function. For example, carbonyl moieties may be
substituted with alkyls, alkenyls, alkynyls, aryls, alkoxy, aminos,
etc. Examples of moieties which contain a carbonyl include
aldehydes, ketones, carboxylic acids, amides, esters, anhydrides,
etc.
[0176] The term "thiocarbonyl" or "thiocarboxy" includes compounds
and moieties which contain a carbon connected with a double bond to
a sulfur atom.
[0177] The term "ether" includes compounds or moieties which
contain an oxygen bonded to two different carbon atoms or
heteroatoms. For example, the term includes "alkoxyalkyl" which
refers to an alkyl, alkenyl, or alkynyl group covalently bonded to
an oxygen atom which is covalently bonded to another alkyl
group.
[0178] The term "ester" includes compounds and moieties which
contain a carbon or a heteroatom bound to an oxygen atom which is
bonded to the carbon of a carbonyl group. The term "ester" includes
alkoxycarboxy groups such as methoxycarbonyl, ethoxycarbonyl,
propoxycarbonyl, butoxycarbonyl, pentoxycarbonyl, etc. The alkyl,
alkenyl, or alkynyl groups are as defined above.
[0179] The term "thioether" includes compounds and moieties which
contain a sulfur atom bonded to two different carbon or hetero
atoms. Examples of thioethers include, but are not limited to
alkthioalkyls, alkthioalkenyls, and alkthioalkynyls. The term
"alkthioalkyls" include compounds with an alkyl, alkenyl, or
alkynyl group bonded to a sulfur atom which is bonded to an alkyl
group. Similarly, the term "alkthioalkenyls" and alkthioalkynyls"
refer to compounds or moieties wherein an alkyl, alkenyl, or
alkynyl group is bonded to a sulfur atom which is covalently bonded
to an alkynyl group.
[0180] The term "hydroxy" or "hydroxyl" includes groups with an
--OH or --O.sup.-.
[0181] The term "halogen" includes fluorine, bromine, chlorine,
iodine, etc. The term "perhalogenated" generally refers to a moiety
wherein all hydrogens are replaced by halogen atoms.
[0182] The terms "polycyclyl" or "polycyclic radical" refer to two
or more cyclic rings (e.g., cycloalkyls, cycloalkenyls,
cycloalkynyls, aryls and/or heterocyclyls) in which two or more
carbons are common to two adjoining rings, e.g., the rings are
"fused rings". Rings that are joined through non-adjacent atoms are
termed "bridged" rings. Each of the rings of the polycycle can be
substituted with such substituents as described above, as for
example, halogen, hydroxyl, alkylcarbonyloxy, arylcarbonyloxy,
alkoxycarbonyloxy, aryloxycarbonyloxy, carboxylate, alkylcarbonyl,
alkoxycarbonyl, alkylaminoacarbonyl, arylalkylaminocarbonyl,
alkenylaminocarbonyl, alkylcarbonyl, arylcarbonyl, arylalkyl
carbonyl, alkenylcarbonyl, aminocarbonyl, alkylthiocarbonyl,
alkoxyl, phosphate, phosphonato, phosphinato, cyano, amino
(including alkyl amino, dialkylamino, arylamino, diarylamino, and
alkylarylamino), acylamino (including alkylcarbonylamino,
arylcarbonylamino, carbamoyl and ureido), amidino, imino,
sulfhydryl, alkylthio, arylthio, thiocarboxylate, sulfates,
alkylsulfinyl, sulfonato, sulfamoyl, sulfonamido, nitro,
trifluoromethyl, cyano, azido, heterocyclyl, alkyl, alkylaryl, or
an aromatic or heteroaromatic moiety.
[0183] The term "heteroatom" includes atoms of any element other
than carbon or hydrogen. Preferred heteroatoms are nitrogen,
oxygen, sulfur and phosphorus.
[0184] The term "ring" means cycloalkyl or aryl as these terms are
used and defined herein.
[0185] The term "prodrug moiety" includes moieties which can be
metabolized in vivo and moieties which may advantageously remain
esterified or otherwise protected in vivo. Preferably, the prodrugs
moieties are metabolized in vivo by esterases or by other
mechanisms to hydroxyl groups or other advantageous groups.
Examples of prodrugs and their uses are well known in the art (See,
e.g., Berge et al. (1977) "Pharmaceutical Salts", J. Pharm. Sci.
66:1-19). The prodrugs can be prepared in situ during the final
isolation and purification of the compounds, or by separately
reacting the purified compound in its free acid form or hydroxyl
with a suitable esterifying agent. Hydroxyl groups can be converted
into esters via treatment with a carboxylic acid. Examples of
prodrug moieties include substituted and unsubstituted, branch or
unbranched lower alkyl ester moieties, (e.g., propionoic acid
esters), lower alkenyl esters, di-lower alkyl-amino lower-alkyl
esters (e.g., dimethylaminoethyl ester), acylamino lower alkyl
esters (e.g., acetyloxymethyl ester), acyloxy lower alkyl esters
(e.g., pivaloyloxymethyl ester), aryl esters (phenyl ester),
aryl-lower alkyl esters (e.g., benzyl ester), substituted (e.g.,
with methyl, halo, or methoxy substituents) aryl and aryl-lower
alkyl esters, amides, lower-alkyl amides, di-lower alkyl amides,
and hydroxy amides.
[0186] It will be noted that the structure of some of the compounds
of this invention includes asymmetric carbon atoms, and thus may
exist as racemic mixtures or as isolated isomeric forms. It is to
be understood accordingly that the isomers arising from such
asymmetry (e.g., all enantiomers and diastereomers) are included
within the scope of this invention, unless indicated otherwise.
Such isomers can be obtained in substantially pure form by
classical separation techniques and by stereochemically controlled
synthesis. Furthermore, the structures and other compounds and
moieties discussed in this application also include all tautomers
thereof.
[0187] The present invention pertains, at least in part, to methods
for treating a blood disorder in a subject by administering to the
subject an effective amount of a compound of the invention (e.g., a
compound of Formula I, II, III or otherwise described herein,
including isolated enantiomers or diastereomers).
[0188] The term "treating" includes curing as well as ameliorating
at least one symptom of the state, disease or disorder, e.g., the
blood disorder. Therefore, prevention of blood disorders or at
least one symptom thereof is also contemplated herein.
[0189] The term "blood disorder" includes disorders which can be
treated, prevented, or otherwise ameliorated by the administration
of a compound of the invention, e.g., a compound of formula I, II,
III or otherwise described herein). A blood disorder is any
disorder of the blood and blood-forming organs. The term blood
disorder includes nutritional anemias (e.g., iron deficiency
anemia, sideropenic dysphasia, Plummer-Vinson syndrome, vitamin
B.sub.12 deficiency anemia, vitamin B.sub.12 deficiency anemia due
to intrinsic factor, pernicious anemia, folate deficiency anemia,
and other nutritional anemias), myelodysplastic syndrome, bone
marrow failure or anemia resulting from chemotherapy, radiation or
other agents or therapies, hemolytic anemias (e.g., anemia due to
enzyme disorders, anemia due to phosphate dehydrogenase (G6PD)
deficiency, favism, anemia due to disorders of glutathione
metabolism, anemia due to disorders of glycolytic enzymes, anemias
due to disorders of nucleotide metabolism and anemias due to
unspecified enzyme disorder), thalassemia .alpha.-thalassemia,
.beta.-thalassemia, .delta..beta.-thalassemia, thalassemia trait,
hereditary persistence of fetal hemoglobin (HPFP), and unspecified
thalassemias), sickle cell disorders (sickle cell anemia with
crisis, sickle cell anemia without crisis, double heterozygous
sickling disorders, sickle cell trait and other sickle cell
disorders), hereditary hemolytic anemias (hereditary spherocytosis,
hereditary elliptocytosis, other hemaglobinopathies and other
specified hereditary hemolytic anemias, such as stomatocyclosis),
acquired hemolytic anemia (e.g., drug-induced autoimmune hemolytic
anemia, other autoimmune hemolytic anemias, such as warm autoimmune
hemolytic anemia, drug-induced non-autoimmune hemolytic anemia,
hemolytic-uremic syndrome, and other non-autoimmune hemolytic
anemias, such as microangiopathic hemolytic anemia); aplastic
anemias (e.g., acquired pure red cell aplasia (erythoblastopenia),
other aplastic anemias, such as constitutional aplastic anemia and
fanconi anemia, acute posthemorrhagic anemic, and anemias in
chronic diseases), coagulation defects (e.g., disseminated
intravascular coagulation (difibrination syndrome)), hereditary
factor VIII deficiency (hemophilia A), hereditary factor IX
deficiency (Christmas disease), and other coagulation defects such
as Von Willebrand's disease, hereditary factor Xi deficiency
(hemophilia C), purpura (e.g., qualitative platelet defects and
Glanzmann's disease), neutropenia, agranulocytosis, functional
disorders of polymorphonuclear neutrophils, other disorders of
white blood cells (e.g., eosinophilia, leukocytosis,
lymophocytosis, lymphopenia, monocytosis, and plasmacyclosis),
diseases of the spleen, methemoglobinemia, other diseases of blood
and blood forming organs (e.g., familial erythrocytosis, secondary
polycythemia, essential thrombocytosis and basophilia),
thrombocytopenia, infectious anemia, hypoproliferative or
hypoplastic anemias, hemoglobin C, D and E disease, hemoglobin
lepore disease, and HbH and HbS diseases, anemias due to blood
loss, radiation therapy or chemotherapy, or thrombocytopenias and
neutropenias due to radiation therapy or chemotherapy,
sideroblastic anemias, myelophthisic anemias, antibody-mediated
anemias, and certain diseases involving lymphoreticular tissue and
reticulohistiocytic system (e.g., Langerhans' cell hystiocytosis,
eosinophilic granuloma, Hand-Schuller-Christian disease,
hemophagocytic lymphohistiocytosis, and infection-associated
hemophagocytic syndrome).
[0190] In one embodiment, the compounds of formula I, II, III or
otherwise described herein stimulate fetal hemoglobin production,
hematopoiesis, erythropoiesis, myelopoiesis and/or neutrophil
production upon administration to a subject for the treatment of a
blood disorder.
[0191] In one embodiment, the compounds of formula I, II, III or
otherwise described are administered to the subject for treatment
of a blood disorder in combination with one or more cytokines. In
one embodiment, the cytokine is selected from the group consisting
of IL-3, GM-CSF, G-CSF, stem cell factor (SCF) and IL-6.
[0192] In the therapeutic methods of the invention, one or more
compounds of the invention may be administered alone to a subject,
or more typically a compound of the invention will be administered
as part of a pharmaceutical composition in mixture with
conventional excipient, i.e., pharmaceutically acceptable organic
or inorganic carrier substances suitable for parenteral, oral or
other desired administration and which do not deleteriously react
with the active compounds and are not deleterious to the recipient
thereof.
[0193] In another embodiment, the invention pertains, at least in
part to a pharmaceutical composition of an effective amount of a
compound of formula I, formula II, formula III, or ##STR43## and
racemates, isolated enantiomers or diastereomers thereof, and a
pharmaceutically acceptable carrier.
[0194] The language "pharmaceutically acceptable carrier" includes
substances capable of being coadministered with the compound(s) of
the invention and which allow both to perform their intended
function, e.g., treat or prevent a blood disorder. Suitable
pharmaceutically acceptable carriers include but are not limited to
water, salt solutions, alcohol, vegetable oils, polyethylene
glycols, gelatin, lactose, amylose, magnesium stearate, talc,
silicic acid, viscous paraffin, perfume oil, fatty acid
monoglycerides and diglycerides, petroethral fatty acid esters,
hydroxymethyl-cellulose, polyvinylpyrrolidone, etc. The
pharmaceutical preparations can be sterilized and if desired mixed
with auxiliary agents, e.g., lubricants, preservatives,
stabilizers, wetting agents, emulsifiers, salts for influencing
osmotic pressure, buffers, colorings, flavorings and/or aromatic
substances and the like which do not deleteriously react with the
active compounds of the invention.
[0195] The compounds of the invention that are basic in nature are
capable of forming a wide variety of salts with various inorganic
and organic acids. The acids that may be used to prepare
pharmaceutically acceptable acid addition salts of the compounds of
the invention that are basic in nature are those that form
non-toxic acid addition salts, i.e., salts containing
pharmaceutically acceptable anions, such as the hydrochloride,
hydrobromide, hydroiodide, nitrate, sulfate, bisulfate, phosphate,
acid phosphate, isonicotinate, acetate, lactate, salicylate,
citrate, acid citrate, tartrate, pantothenate, bitartrate,
ascorbate, succinate, maleate, gentisinate, fumarate, gluconate,
glucaronate, saccharate, formate, benzoate, glutamate,
methanesulfonate, ethanesulfonate, benzenesulfonate,
p-toluenesulfonate and palmoate [i.e.,
1,1'-methylene-bis-(2-hydroxy-3-naphthoate)] salts. Although such
salts must be pharmaceutically acceptable for administration to a
subject, e.g., a mammal, it is often desirable in practice to
initially isolate a compound of the invention from the reaction
mixture as a pharmaceutically unacceptable salt and then simply
convert the latter back to the free base compound by treatment with
an alkaline reagent and subsequently convert the latter free base
to a pharmaceutically acceptable acid addition salt. The acid
addition salts of the base compounds of this invention are readily
prepared by treating the base compound with a substantially
equivalent amount of the chosen mineral or organic acid in an
aqueous solvent medium or in a suitable organic solvent, such as
methanol or ethanol. Upon careful evaporation of the solvent, the
desired solid salt is readily obtained. The preparation of other
compounds of the invention not specifically described in the
foregoing experimental section can be accomplished using
combinations of the reactions described above that will be apparent
to those skilled in the art.
[0196] The compounds of the invention that are acidic in nature are
capable of forming a wide variety of base salts. The chemical bases
that may be used as reagents to prepare pharmaceutically acceptable
base salts of those compounds of the invention that are acidic in
nature are those that form non-toxic base salts with such
compounds. Such non-toxic base salts include, but are not limited
to those derived from such pharmaceutically acceptable cations such
as alkali metal cations (e.g., potassium and sodium) and alkaline
earth metal cations (e.g., calcium and magnesium), ammonium or
water-soluble amine addition salts such as
N-methylglucamine-(meglumine), and the lower alkanolammonium and
other base salts of pharmaceutically acceptable organic amines. The
pharmaceutically acceptable base addition salts of compounds of the
invention that are acidic in nature may be formed with
pharmaceutically acceptable cations by conventional methods. Thus,
these salts may be readily prepared by treating the compound of the
invention with an aqueous solution of the desired pharmaceutically
acceptable cation and evaporating the resulting solution to
dryness, preferably under reduced pressure. Alternatively, a lower
alkyl alcohol solution of the compound of the invention may be
mixed with an alkoxide of the desired metal and the solution
subsequently evaporated to dryness.
[0197] The compounds of the invention and pharmaceutically
acceptable salts thereof can be administered via either the oral,
parenteral or topical routes. In general, these compounds are most
desirably administered in effective dosages, depending upon the
weight and condition of the subject being treated and the
particular route of administration chosen. Variations may occur
depending upon the species of the subject being treated and its
individual response to said medicament, as well as on the type of
pharmaceutical formulation chosen and the time period and interval
at which such administration is carried out.
[0198] The pharmaceutical compositions of the invention may be
administered alone or in combination with other known compositions
for treating blood disorders in a subject, e.g., a mammal.
Preferred mammals include cats, dogs, pigs, rats, mice, monkeys,
chimpanzees, baboons and humans. In one embodiment, the subject is
suffering from a blood disorder. In another embodiment, the subject
is at risk of suffering from a blood disorder.
[0199] The language "in combination with" a known composition is
intended to include simultaneous administration of the composition
of the invention and the known composition, administration of the
composition of the invention first, followed by the known
composition and administration of the known composition first,
followed by the composition of the invention. Any of the
therapeutically composition known in the art for treating blood
disorders can be used in the methods of the invention.
[0200] The compounds of the invention may be administered alone or
in combination with pharmaceutically acceptable carriers or
diluents by any of the routes previously mentioned, and the
administration may be carried out in single or multiple doses. For
example, the novel therapeutic agents of this invention can be
administered advantageously in a wide variety of different dosage
forms, i.e., they may be combined with various pharmaceutically
acceptable inert carriers in the form of tablets, capsules,
lozenges, troches, hard candies, powders, sprays, creams, salves,
suppositories, jellies, gels, pastes, lotions, ointments, aqueous
suspensions, injectable solutions, elixirs, syrups, and the like.
Such carriers include solid diluents or fillers, sterile aqueous
media and various non-toxic organic solvents, etc. Moreover, oral
pharmaceutical compositions can be suitably sweetened and/or
flavored. In general, the therapeutically-effective compounds of
this invention are present in such dosage forms at concentration
levels ranging from about 5.0% to about 70% by weight.
[0201] For oral administration, tablets containing various
excipients such as microcrystalline cellulose, sodium citrate,
calcium carbonate, dicalcium phosphate and glycine may be employed
along with various disintegrants such as starch (and preferably
corn, potato or tapioca starch), alginic acid and certain complex
silicates, together with granulation binders like
polyvinylpyrrolidone, sucrose, gelatin and acacia. Additionally,
lubricating agents such as magnesium stearate, sodium lauryl
sulfate and talc are often very useful for tabletting purposes.
Solid compositions of a similar type may also be employed as
fillers in gelatin capsules; preferred materials in this connection
also include lactose or milk sugar as well as high molecular weight
polyethylene glycols. When aqueous suspensions and/or elixirs are
desired for oral administration, the active ingredient may be
combined with various sweetening or flavoring agents, coloring
matter or dyes, and, if so desired, emulsifying and/or suspending
agents as well, together with such diluents as water, ethanol,
propylene glycol, glycerin and various like combinations
thereof.
[0202] For parenteral administration (including intraperitoneal,
subcutaneous, intravenous, intradermal or intramuscular injection),
solutions of a therapeutic compound of the present invention in
either sesame or peanut oil or in aqueous propylene glycol may be
employed. The aqueous solutions should be suitably buffered
(preferably pH greater than 8) if necessary and the liquid diluent
first rendered isotonic. These aqueous solutions are suitable for
intravenous injection purposes. The oily solutions are suitable for
intraarticular, intramuscular and subcutaneous injection purposes.
The preparation of all these solutions under sterile conditions is
readily accomplished by standard pharmaceutical techniques well
known to those skilled in the art. For parenteral application,
examples of suitable preparations include solutions, preferably
oily or aqueous solutions as well as suspensions, emulsions, or
implants, including suppositories. Therapeutic compounds may be
formulated in sterile form in multiple or single dose formats such
as being dispersed in a fluid carrier such as sterile physiological
saline or 5% saline dextrose solutions commonly used with
injectables.
[0203] Additionally, it is also possible to administer the
compounds of the present invention topically when treating
inflammatory conditions of the skin. Examples of methods of topical
administration include transdermal, buccal or sublingual
application. For topical applications, therapeutic compounds can be
suitably admixed in a pharmacologically inert topical carrier such
as a gel, an ointment, a lotion or a cream. Such topical carriers
include water, glycerol, alcohol, propylene glycol, fatty alcohols,
triglycerides, fatty acid esters, or mineral oils. Other possible
topical carriers are liquid petrolatum, isopropylpalmitate,
polyethylene glycol, ethanol 95%, polyoxyethylene monolauriate 5%
in water, sodium lauryl sulfate 5% in water, and the like. In
addition, materials such as anti-oxidants, humectants, viscosity
stabilizers and the like also may be added if desired.
[0204] For enteral application, particularly suitable are tablets,
dragees or capsules having talc and/or carbohydrate carrier binder
or the like, the carrier preferably being lactose and/or corn
starch and/or potato starch. A syrup, elixir or the like can be
used wherein a sweetened vehicle is employed. Sustained release
compositions can be formulated including those wherein the active
component is derivatized with differentially degradable coatings,
e.g., by microencapsulation, multiple coatings, etc.
[0205] It will be appreciated that the actual preferred amounts of
active compounds used in a given therapy will vary according to the
specific compound being utilized, the particular compositions
formulated, the mode of application, the particular site of
administration, etc. Optimal administration rates for a given
protocol of administration can be readily ascertained by those
skilled in the art using conventional dosage determination tests
conducted with regard to the foregoing guidelines.
[0206] In general, compounds of the invention for treatment can be
administered to a subject in dosages used in prior therapies. For
example, a suitable effective dose of one or more compounds of the
invention will be in the range of from 0.01 to 100 milligrams per
kilogram of body weight of recipient per day, preferably in the
range of from 0.1 to 50 milligrams per kilogram body weight of
recipient per day, more preferably in the range of 1 to 20
milligrams per kilogram body weight of recipient per day. The
desired dose is suitably administered once daily, or several
sub-doses, e.g. 2 to 5 sub-doses, are administered at appropriate
intervals through the day, or other appropriate schedule.
[0207] It will also be understood that normal, conventionally known
precautions will be taken regarding the administration of the
compounds of the invention generally to ensure their efficacy under
normal use circumstances. Especially when employed for therapeutic
treatment of humans and animals in vivo, the practitioner should
take all sensible precautions to avoid conventionally known
contradictions and toxic effects.
[0208] Furthermore, the invention also pertains to the use of a
compound of formula I, II, III or a compound otherwise described
herein for the preparation of a medicament. The medicament may
include a pharmaceutically acceptable carrier and the compound is
an effective amount, e.g., an effective amount to treat a blood
disorder.
EXEMPLIFICATION OF THE INVENTION
Example 1
Identification of Small Molecule Inducers of Fetal Hemoglobin
[0209] Small molecule inducers of fetal hemoglobin were identified
using computer modeling techniques.
[0210] The pharmacophore was constructed with the TFIT module of
the FLO molecular modeling software. It was assumed that the
carboxylic acids would bind to the receptor in an analogous
fashion, and therefore, the superposition of the carboxylic oxygens
was biased by imposing a 5 kJ superimposition energy constraint.
Five hundred iterations of TFIT were used in the calculations.
[0211] TFIT produced an ensemble of low energy superimposition. The
superimposition with the tightest overlay was taken to be the
initial pharmacophore template. This pharmacophore was tested to
see if it could distinguish between active and inactive compounds.
TFIT was first used to determine the best match between the
pharmacophore and four compounds which had been identified as
inactive in the .beta./.gamma.-globin promoter driven reporter gene
assay in previous studies.
[0212] The TFIT was next used to determine how well five additional
compounds, which were active in the .beta./.gamma.-globin promoter
driven reporter gene assay would match the template. The template
was used to design and select new compounds for testing. Compounds
were selected from available compound data bases and evaluated by
fitting them onto the template with TFIT.
[0213] The compounds generated from the modelling were tested in
the .beta./.gamma.-globin promoter driven reporter gene assay and
were found to have statistically significant activity in the assay
(Table 1). All of the compounds shown in Table 1 had a % y globin
promoter induction (above untreated control) of between about
100%-200%. TABLE-US-00001 TABLE 1 Compound Code Structure A
##STR44## B ##STR45## C ##STR46## D ##STR47## E ##STR48## H
##STR49## I ##STR50## J ##STR51##
[0214] Next, a "pseudo" receptor was constructed around the
pharmacophore by refining the original pharmacophore template. A
new template was constructed by adding two of the most active new
compounds. The compounds were selected primarily for the additional
structural information that they contained.
[0215] The "pseudo" binding site was construction using FLO. This
"pseudo" binding site was composed of functional groups selected to
form hydrogen bonds with the ligands, and functional groups that
would mimic the hydrophobic surface of the binding site. A
guanidinium group was selected form hydrogen bonds with the acidic
groups of the ligands. A pyrrole group was used to mimic the
binding site hydrogen bond donors. These groups were positioned
around the template molecules and anchored to the chemically
complimentary ligand atoms with a 10 kJ constraint. The "pseudo"
program of FLO automatically filled the remaining volume with
propane to mimic the binding site's hydrophobic surface. This
structure was next subject to several rounds of dynamics.
[0216] Once the template atoms were removed, the shell of propanes,
the pyrrole group and the guanidinium groups represented the
receptor binding site. To ensure a moderate amount of binding site
flexibility, the atoms of the binding site were allowed to move
with a molecular mechanics force field and an additional flat well
constraint [radius 0.5 .ANG., quadratic penalty 20 kJ/.ANG..sup.2]
was imposed.
[0217] To test the "pseudo" binding site, twenty compounds (14
active and 6 inactive in the .beta./.gamma.-globin promoter driven
reporter gene assay) were docked into the binding site model using
the docking module SDOCK+ of FLO+. For each docked conformation,
FLO+ computed a predicted binding free energy using an empirical
scoring function consisting of contact energy, hydrogen bonding
energy, polar desolvation, bumping, internal energy and entropy.
For the most active compounds, the predicted free energy (reported
as pI or the -logK.sub.i), fell between 6.6 and 6.9, with hydrogen
bonding energies between 7.4 and 8.9 kJ/mol. The pI values for the
four inactive compounds ranged from 5.6 to 6.2 with hydrogen
bonding energies between 5.6 and 7.0 kJ/mol. A combination of the
pI and the hydrogen bonding energy was used to distinguish between
active and inactive compounds.
[0218] Compounds for screening were then selected from a database
of 13,000 commercially available compounds. Only molecules with an
acid group and less that 24 heavy atoms were chosen, resulting in
630 compounds. These compounds were docked into the binding site
using SDOCK+. The binding modes were scored using FLO+ and the best
10 conformations for each compound were retained for visual
inspection. TABLE-US-00002 TABLE 2 % .UPSILON. globin promoter
Compound induction (above Code Structure untreated control F
##STR52## *** K ##STR53## * M ##STR54## ** N ##STR55## *** O
##STR56## * P ##STR57## *** Q ##STR58## ** R ##STR59## *** S
##STR60## ** T ##STR61## ** V ##STR62## ** W ##STR63## ** X
##STR64## ** Y ##STR65## ** Z ##STR66## ** AA ##STR67## ** AB
##STR68## ** AC ##STR69## ** AD ##STR70## ** AE ##STR71## ** AF
##STR72## * AG ##STR73## ** AH ##STR74## ** AI ##STR75## ** AJ
##STR76## ** AK ##STR77## ** Using the pl and hydrogen bonding
energy as criteria, 30 compounds were selected for in vitro
testing. The scores for these compounds ranged for pl 5.1 (hydrogen
bonding energy of -8.1 kJ/mol) to pl 8.8 (hydrogen bonding of 9.4
kJ/mol). Twenty six of these compounds were acquired and tested.
Table 2 shows the results of the .beta./.UPSILON.-globin promoter
driven reporter gene assay for the twenty-six compounds. `*`
indicates a 80-100% increase; `**` indicates a 100%- #200%
increase; and `***` indicates an over 200% increase of .UPSILON.
globin promoter induction over untreated controls.
Example 2
IN Vitro Stimulation of Fetal Globin mRNA Expression
[0219] This example demonstrates that the test compounds predicted
to be active by reporter gene assays and molecular modeling produce
a significant increase in fetal (gamma) globin mRNA in cells
cultured in vitro. Furthermore, the concentrations required were
significantly lower (5-40 micromolar) than concentrations required
for prior generation inducers (100-200 micromolar), making these
compounds more suitable for therapeutic and pharmacologic
compositions
[0220] .gamma.-globin mRNA was analyzed in control and treated
erythroid colonies cultured from cord blood, by RT-PCR.
[0221] Induction (increase) in fetal globin mRNA compared to
untreated control levels with each compound is shown in Table 3
below. The R enantiomer of compound Y demonstrated fetal globin
inducing action, whereas the S enantiomer did not induce fetal
globin in 2 of (the same) 3 cultures. TABLE-US-00003 TABLE 3
Increase in Fetal Globin mRNA Concentration Mean change required,
above control, No. of Positive Compound micromolar % responses M 5
70 4/6 P 20 86 6/6 2-methyl-1- 5 46 6/6 benzofuran-4- carboxylic
acid V 5 50 6/6 W 40 368 5/6 3-(5- 30 50 chlorothien-3- yl)acrylic
acid Y racemic mix 5 20 1/3 Y + R 5 67 2/3 Y - S 5 -2 1/3
[0222] These levels of fetal globin induction are higher than the
induction by previously reporter inducing agents, and occur at
lower concentrations, i.e., these agents have higher potency.
[0223] In a related study, the relative luciferase reporter gene
induction, .gamma.-globin gene induction, and F cell production was
tested for several of the candidate compounds. The relative in
vitro .gamma.-globin gene reporter stimulation for the tested
compounds was as follows: compound P>M>W=R=Y. The relative in
vitro .gamma.-globin gene induction for the tested compounds was as
follows: P>R=M>Y>W>. The relative in vitro F-cell
production was as follows: W>Y>R>M>P. The relative
potency for F-cell production was as follows:
R>M>Y>P>W.
Example 3
Effects on Erythroid and Myeloid Cell Growth In Vitro
[0224] This example demonstrates that the test compounds predicted
to be active by reporter gene assays and molecular modeling produce
a significant increase in numbers of erythroid and myeloid colonies
or proportion of cells expressing fetal globin in in vitro cultured
cells from a variety of sources under a variety of culture
conditions. Similar to other in vitro tests described herein, the
concentrations required for these biological effects were
significantly lower (5-40 micromolar) than concentrations required
for prior generation inducers (100-200 micromolar), making these
compounds more suitable for therapeutic and pharmacologic
compositions.
[0225] Compounds predicted in the molecular model to be
.gamma.-globin inducers were evaluated in a series of assays for
activity in 1) stimulating activity from the fetal globin gene
promoter, (the action which can ameliorate sickle cell disease and
beta thalassemia), and 2) for any effects on stimulating erythroid
or myeloid cell growth and proliferation, the action which can
treat blood cell deficiencies.
[0226] Erythroid burst-forming units (BFU-E) (erythroid
progenitors) and colony-forming units granulocyte-macrophage
(CFU-GM) (myeloid colonies) were cultured in semi-solid or in
two-phase suspension media, with or without hematopoietic growth
factors at high levels (e.g., erythropoietin at 3 U/ml) or reduced
levels (BFU-E cell proliferation was evaluated by enumeration in
colonies developing in the presence of reduced amounts of
erythropoietin (0.5 U/ml) rather than 3 U (or 3000 mU)/ml, which is
standard for these experimental systems), from cord blood or the
peripheral blood of several types of humans. Experiments were
carried out on samples derived from 1) .beta.-thalassemia patients
who expressed variable levels of fetal globin at baseline (and in
untreated control cultures) and represent a variety of potential
individual responses, 2) from normal umbilical cord blood samples,
which express 40-50% fetal globin at baseline (and in untreated
control cultures), 3) from CD34+ cells isolated from normal adult
peripheral blood, which expressed low levels of fetal globin in
untreated control cultures and at baseline; and 4) from peripheral
blood of patients with sickle cell disease who were receiving
treatment with hydroxyurea. These biological samples provide a
range of potential difficulty in stimulating fetal globin
expression in human patients.
[0227] Experiment 1: Hematopoietic colonies were enumerated with or
without (+/-) the test compounds, and compared to colonies which
developed in the presence of hematopoietic growth factors alone
from the same subject (untreated control colonies), and proportions
of cells expressing fetal globin (F-cells) were analyzed by FACScan
analysis. The compounds M, N, P, R, T,
2-methyl-1-benzofuran-4-carboxylic acid, V, W, X, Y, Z, F and
3-(5-chlorothien-3-yl)acrylic acid, all resulted in significant
increases in the proportion of F-cells above the percentage of
F-cells in untreated controls, at the same low concentrations as
required for fetal globin mRNA induction and increases in erythroid
colony numbers with the compounds.
[0228] Experiment 2: Erythroid colonies were cultured from patients
with beta thalassemia, either without any test compounds (Control),
only an optimal panel of hematopoietic growth factors, or with one
of the test compounds M, P, T, 2-methyl-1-benzofuran-4-carboxylic
acid, V, W, X, Y, AK, AC, or AI. All of the listed test compounds
increased the number of colonies as compared to the matched control
cultures that were grown with an optimal panel of growth factors
alone. Colony numbers were increased above the control cultures (%
BFU-E/Control) by anywhere from 25% to 230%. Each of the listed
test compounds was tested in at least 5 different patients' blood
and the differences were statistically signficant.
[0229] In a related study, BFU-E cultured from cord blood, was
tested with or without compound M, P, R, T, W, Y, Z, AI, F, or
3-(5-chlorothien-3-yl)acrylic acid. The test compounds all induced
increased numbers of colonies as compared to the controls. In this
study the test compounds resulted in 50-250% more colonies than in
control cultures from the same source.
[0230] Experiment 3: Representative novel compounds M, W, X, Y, and
AI also stimulated production of myeloid colonies compared to
control, untreated myeloid colonies from the same individual.
Control colonies were established in cultures with no added growth
factors to support myelopoiesis, in Iscove's Modified Dulbecco's
Media (IMDM) methylcellulose media with charcoal-absorbed fetal
bovine serum, beta mercaptoethanol, BSA. A 30-75% increase in de
novo myeloid colonies was observed in cord blood cultured with the
novel compounds at the same concentrations as required for
increases in erythroid colonies.
[0231] Experiment 4: Erythroid cells cultured from adult blood are
low-HbF expressing and are the most difficult to alter with regard
to globin expression. This experiment analyzed erythroid cells
cultured from adult blood in the presence or absence of
representative test compounds.
[0232] Erythroid cells were generated by culturing purified CD34+
cells in Flt-3 ligand, stem cell factor (SCF) and IL-3 for seven
days followed by growth in EPO for 14 days. For treatment, cells
were cultured as above in a T75 flask then split into multiple
flasks on day 8 and treatment with the test compounds was begun.
Compound P or other test compounds were diluted from stock
solutions with the microliter volumes of stock added to each
culture flask for final working concentrations. Cells were
enumerated every two days by hemacytometer count. RNA was harvested
from 10.sup.6 cells every two days using RNeasy Plus Mini Kit
(Qiagen) and qRT-PCR performed using IQ SybrGreen Supermix on an
Opticon Monitor instrument (MJ Research). Samples were assayed in
triplicate and raw data from the instrument was analyzed using a
method suggested by Larionov et al. with beta-actin and G3PD
assayed as controls (housekeeping genes). Separation of hemoglobins
was performed by cation exchange HPLC using a 35.times.4.6 mm; 3 mm
PolyCAT A column (Nest Group) as described previously by Cheryl
Rognerud and Ching-Nan Ou, and outlined by the column
manufacturer.
[0233] QPCR primers were designed using known sequences for
.alpha.-globin, .beta.-globin, .gamma.-globin, .beta.-actin and
B3PD. Primers were designed to span at least one exon.
[0234] The results showed that erythroid cells peaked at day 12-14
of the erythroid phase. Cell counts were increased with 20 microM
compound P as compared to control cells cultured with
erythropoietin alone. Comparison of compound P at different doses
showed that there was no effect on production/expression of alpha
globin, but there was a reciprocal decrease in production of beta
globin concomitantly with increases in fetal globin mRNA expression
at the same doses. The activity was specific for inducing gamma
(fetal) globin with reciprocal decrease in beta globin. Hemoglobin
F protein levels increased in a compound P dose-dependent manner
with a 19-fold increase in expression of fetal globin produced at
100 micromolar compound P as compared to untreated control cultures
from the same subject with just one cycle of erythroid
differentiation.
[0235] Experiment 5: In an additional study, comparative growth of
erythroid (red blood cell) colonies cultured from cord blood
samples was evaluated with the addition of growth factors alone or
with the addition of test compounds.
[0236] Comparative growth of erythroid colonies cultured from cord
blood samples under conditions with standard growth factors alone
(Control), or with addition of test compounds or arginine butyrate
(AB) (AB results in reduced colony numbers). Control and test
cultures were established from the same samples with the same
growth factors (EPO and II-3) with addition of test compounds. The
compounds tested were M, P, R, T, W, Y, Z, AI, F and
3-(5-chlorothien-3-yl)acrylic acid. Two compounds previously shown
as positive proliferative agents were included for comparison. All
compounds tested, except compound AI and
3-(5-chlorothien-3-yl)acrylic acid, resulted in an increase in red
blood cell colony numbers by at least 25% above control numbers and
was statistically significant. These results indicate that the test
compounds stimulate red blood cell production even in conditions of
maximal growth factors and in the absence of anemia.
[0237] Experiment 6: Erythroid colonies from the peripheral blood
of 4 patients with sickle cell disease who were receiving treatment
with Hydroxyurea (HU), a chemotherapeutic agent which suppresses
marrow growth and reduces red blood cell production, were cultured
alone or with added test compounds M, P,
2-methyl-1-benzofuran-4-carboxylic acid, V, W, Y (racemic mixture)
or AI. L-arginine was added as a neutral control, and had no effect
on numbers of erythroid colonies. Addition of arginine butyrate
(AB), phenylacetate (PA), and hydroxyurea (HU) resulted in
decreased numbers of erythroid colonies compared to control
conditions.
[0238] As summarized in Table 5 below, addition of the test
compounds M, P, 2-methyl-1-benzofuran-4-carboxylic acid, V, W, Y
(racemic mixture) or AI (at the concentrations shown in the Table
5) resulted in an increase in numbers of erythroid colonies of at
least 25% above control conditions. Compounds marked with a * are
significantly different. 2-methyl-1-benzofuran-4-carboxylic acid
and compound V require additional samples for statistical
evaluation, but had positive effects in 3/3 different patients'
cultures. These findings indicate that the test compounds stimulate
erythroid cell production particularly in conditions of anemia and
bone marrow suppression. TABLE-US-00004 TABLE 5 Erythroid Colony
Growth from Sickle Cell Patients +/- Test Compounds % Change from
control conditions with growth factors alone Number with
Concentration, % Change effects/ Compound micromolar from Control
No effect P-value L-arginine 150 -3 3/5 0.6, not (neutral
significant control) AB (arginine 100 -29 5/6 0.019 butyrate) PA
100 -10 5/7 0.2 Phenylacetate M* 1 +58 5/5 0.035 P* 100 +32 7/7
0.016 2-methyl-1- 10 +51 3/3 0.1 benzofuran- 4-carboxylic acid V 10
+35 3/3 0.09 W* 40 +47 5/5 0.048 Y racemic 50 +47 6/7 0.024 mix*
Al* 30 +51 4/5 0.04 Hydroxyurea 20 -33 2/2 0.58
[0239] In summary, the experiments described in this example show
that representative test compounds produce a significant increase
in numbers of erythroid and myeloid colonies or proportion of cells
expressing fetal globin in in vitro cultured cells derived from a
variety of sources under culture conditions relevant to blood
conditions including anemia, sickle cell anemia and beta
thalassamia.
Example 4
In Vivo Efficacy in a Non-Human Primate Model
[0240] Compounds P, Y and W were evaluated in a non-human primate
model to evaluate functional activity of these candidate compounds
in stimulating either fetal globin expression or production of
blood cells. As described below, these studies demonstrated potent
in vivo activity of the compounds for inducing fetal globin
expression and production of blood cells.
[0241] Juvenile baboons were catheterized with venous and arterial
catheters, and were phlebotomized a set amount of blood daily to
maintain anemia with a hemoglobin level of 7.0-7.5 g/dl, from a
baseline normal hemoglobin level of (12-13 g/dl). Normal saline was
infused to replace the amount of blood withdrawn. This degree of
phlebotomy exchanges the blood volume every 10-20 days.
[0242] Fetal globin mRNA was analyzed at baseline anemia before and
following administration of test compounds. Globin chain protein
synthesis confirmed the mRNA findings.
[0243] Following establishment of the stable level of anemia, a
test compound was administered once/day, either intravenously or
orally, and blood was withdrawn through the arterial catheter for
analysis of globin mRNA or for analysis of the test compound in the
plasma by HPLC-MS. Pharmacokinetic profiles were established from
the plasma levels and oral bioavailability (comparing area under
the curve between IV and oral plasma levels) was determined for the
test compounds.
[0244] Pharmacokinetic profiles of compounds M (100 mg/kg), P (25
mg/kg), 2-methyl-1-benzofuran-4-carboxylic acid (100 mg/kg), W (100
mg/kg), and Y (10 mg/kg) indicated that these compounds persist
well above therapeutic levels for >8 hours after tolerable
single oral doses. Human equivalent doses are 20-50% of doses in
baboons. Unusually low doses of compound P (25 mg/kg) and of
compound Y (10 mg/kg), respectively, were required. Previously
reported compounds typically require 100-500 mg/kg/dose for
induction of fetal globin. Compound Y in particular also does not
produce an undesirable high initial burst level and persists for
greater than 24 hours. The human equivalent doses are 10-20% of the
baboon dose. Thus for compound Y, the human equivalent dose would
be 1-2 mg/kg, a dose highly favorable for a pharmaceutical
composition.
[0245] Compound P and compound Y treatment in anemic baboons
induced 3 to 6-fold .gamma.-globin mRNA expression. Prior
generation compounds typically induce fetal globin mRNA only by
2-fold.
[0246] In a further in vivo study, compounds AK, Y and M were
evaluated in a baboon and demonstrated significant increases in
fetal globin expression at tolerable low doses. As shown in the
previous Examples, these 3 compounds all stimulated erythroid cell
proliferation in vitro and have favorable pharmacokinetic profiles
after oral administration. The compounds were administered to an
anemic baboon and studied for pharmacodynamic effects on induction
of fetal globin, which is known to ameliorate the pathology of
sickle cell anemia and beta thalassemia.
[0247] In anemic Baboon 5002, baseline fetal globin mRNA, assayed
by RNAse protection, was 26% of non-alpha globin. With treatment
with compound Y at 5 mg/kg/dose, fetal globin mRNA increased to 48%
of non-alpha globin, an 85% increase over the baseline level.
Treatment with compound AK at 50 mg/kg induced a 51% fetal globin
mRNA increase, a 96% increase over the baseline level. Compound M
treatment at 40 mg/kg induced a 67% fetal globin mRNA increase, a
257% increase over the baseline level. The human equivalent dose is
20% of these doses, or 1 mg/kg, 10 mg/kg, or 8 mg/kg respectively.
Thus, these 3 compounds are highly suitable for pharmaceutical
compositions at doses that human patients can readily tolerate.
[0248] In a related study, compound P was evaluated in a
phlebotomized, anemic baboon showing no baseline expression of
fetal globin and 100% of globin expression as beta globin.
Treatment with compound P resulted in an induction of 7% fetal
globin and a concomitant reduction in beta globin expression.
[0249] These findings show that these compounds are excellent
candidates for treatment of the beta hemoglobinopathies and
thalassemias.
Example 5
Induction of Fetal Globin Expression in an In Vivo Transgenic Mouse
Model
[0250] This example demonstrates that compound W increases
expression of human fetal globin mRNA expression and hematocrit in
a transgenic mouse model.
[0251] Transgenic mice containing the human fetal globin gene and a
micro-Locus Control Region (LCR) received no treatment (controls)
or were treated with test compounds and .gamma.-globin mRNA was
analyzed by RT-PCR. Compound W was administered to mice transgenic
for the micro-LCR-201 gamma-globin gene promoter, by IP injection
once/day for 5 days/week for 2 weeks, and gamma-globin mRNA was
assayed by qPCR. Hematocrit was analyzed once per week. A
significant difference in expression of human fetal globin mRNA and
in hematocrit between Control and compound W-treated mice was found
in initial experiments as summarized in Table 4 below.
TABLE-US-00005 TABLE 4 .gamma.-Globin Induction by Compound W in
Transgenic Mice .gamma.-globin mRNA/S16 Study Treatment (mean
relative units) Hematocrit None 1508 51 +/- 0.45 Compound W 3418 57
+/- 3 p-value, paired t-test <0.05 <0.05
Example 6
Synthesis of Compound Y Enantiomers
[0252] ##STR78##
[0253] Cesium thiobenzoate. To a solution of thiobenzoic acid (50.0
g, 361.8 mmol) in methanol (362 mL) was added Cs.sub.2CO.sub.3 (130
g, 398 mmol) in portions over 5 min. The resulting mixture was
stirred 10 min until all solids were dissolved. The resulting
solution was concentrated on the rotovap. The solid residue was
diluted with 500 mL of acetone and the white solid (CsHCO.sub.3)
was filtered off. This process was repeated two times to ensure all
CsHCO.sub.3 was removed. The acetone was then concentrated to
afford cesium thiobenzoate (Strijtveen, B.; Kellogg, R. M. J. Org.
Chem. 1986, 51, 3664) as a yellow solid (81.5 g, 83%); .sup.1H NMR
(300 MHz, CD.sub.3OD) .delta. 7.25-7.38 (m, 3H), 8.09 (dd, J=1.4,
8.2 Hz, 2H). ##STR79##
[0254] (R)-2-Benzoylthio-3-methylbutanoic acid. To a solution of
(S)-(-)-2-bromo-3-methylbutyric acid (4.20 g, 23.2 mmol) in DMF (41
mL) was added cesium thiobenzoate (6.08 g, 22.5 mmol). The mixture
was stirred at rt for 20 h. The resulting solution was diluted with
ether (200 mL) and washed with H.sub.2O (4.times.40 mL). The
ethereal layer was dried (Na.sub.2SO.sub.4), and concentrated. The
crude residue was recrystallized from hexanes to afford
(R)-2-benzoylthio-3-methylbutanoic acid as a pale yellow solid
(4.05 g, 75%); .sup.1H NMR (300 MHz, CDCl.sub.3) .delta. 1.09 (d,
J=7.4 Hz, 3H), 1.11 (d, J=7.4 Hz, 3H), 2.40 (m, 1H), 4.37 (d, J=5.9
Hz, 1H), 7.45 (t, J=7.4 Hz, 2H), 7.57 (t, J=7.4 Hz, 1H), 7.97 (d,
J=7.4 Hz, 2H), 11.9 (br s, 1H); .sup.13C NMR (75 MHz, CDCl.sub.3)
.delta. 19.8, 20.6, 30.5, 53.1, 127.6, 128.8, 133.9, 136.3, 177.9,
190.3; IR (neat) 3100, 2967, 1709, 1669 cm.sup.-1;
[.alpha.].sub.D.sup.22=+95.6 (C.sub.1, CH.sub.2Cl.sub.2). All
spectral data was identical to that previously published
(Strijtveen, B.; Kellogg, R. M. J. Org. Chem. 1986, 51, 3664).
##STR80##
[0255] (S)-2-Benzoylthio-3-methylbutanoic acid. To a solution of
(R)-(+)-2-bromo-3-methylbutyric acid (4.20 g, 23.2 mmol) in DMF (41
mL) was added cesium thiobenzoate (6.08 g, 22.5 mmol). The mixture
was stirred at rt for 20 h. The resulting solution was diluted with
ether (200 mL) and washed with H.sub.2O (4.times.40 mL). The
ethereal layer was dried (Na.sub.2SO.sub.4), and concentrated. The
crude residue was recrystallized from hexanes to afford
(S)-2-benzoylthio-3-methylbutanoic acid as a pale yellow solid
(3.89 g, 72%); .sup.1H NMR (300 MHz, CDCl.sub.3) .delta. 1.09 (d,
J=7.4 Hz, 3H), 1.11 (d, J=7.4 Hz, 3H), 2.40 (m, 1H), 4.37 (d, J=5.9
Hz, 1H), 7.45 (t, J=7.4 Hz, 2H), 7.57 (t, J=7.4 Hz, 1H), 7.97 (d,
J=7.4 Hz, 2H), 12.0 (br s, 1H); .sup.13C NMR (75 MHz, CDCl.sub.3)
.delta. 19.8, 20.6, 30.5, 53.1, 127.6, 128.8, 133.9, 136.3, 177.9,
190.3; IR (neat) 3100, 2967, 1709, 1669 cm.sup.-1;
[.alpha.].sub.D.sup.22=-94.2 (c 1, CH.sub.2Cl.sub.2). ##STR81##
[0256] (R)-2-Mercapto-3-methylbutanoic acid. To a solution of
(R)-2-benzoylthio-3-methylbutanoic acid (4.05 g, 17.0 mmol) in
CH.sub.2Cl.sub.2 (68 mL) was added 3 M aqueous NH.sub.4OH (68 mL).
The mixture was stirred at rt for 3 h. The resulting solution was
diluted with 2 M aqueous KOH (68 mL) and washed with
CH.sub.2Cl.sub.2 (6.times.70 mL) to remove the benzamide. The
aqueous layer was then acidified to pH 2 with concentrated aqueous
HCl and extracted with CH.sub.2Cl.sub.2 (4.times.70 mL). The
organic extracts were dried (Na.sub.2SO.sub.4) and concentrated to
afford (R)-2-mercapto-3-methylbutanoic acid as a white solid (2.10
g, 92%); .sup.1H NMR (300 MHz, CDCl.sub.3) .delta. 1.05 (d, J=6.7
Hz, 3H), 1.09 (d, J=6.7 Hz, 3H), 1.96 (d, J=9.7 Hz, 1H), 2.07 (m,
1H), 3.13 (dd, J=8.1, 9.7 Hz, 1H); .sup.13C NMR (75 MHz,
CDCl.sub.3) .delta. 19.3, 20.8, 32.6, 48.8, 179.8; IR (neat) 3100,
2966, 1705 cm.sup.-1; [.alpha.].sub.D.sup.22=+41.0 (c 1, ether).
##STR82##
[0257] (S)-2-Mercapto-3-methylbutanoic acid. To a solution of
(S)-2-benzoylthio-3-methylbutanoic acid (3.89 g, 16.3 mmol) in
CH.sub.2Cl.sub.2 (65 mL) was added 3 M aqueous NH.sub.4OH (65 mL).
The mixture was stirred at rt for 3 h. The resulting solution was
diluted with 2 M aqueous KOH (65 mL) and washed with
CH.sub.2Cl.sub.2 (6.times.65 mL) to remove the benzamide. The
aqueous layer was then acidified to pH 2 with concentrated aqueous
HCl and extracted with CH.sub.2Cl.sub.2 (4.times.65 mL). The
organic extracts were dried (Na.sub.2SO.sub.4) and concentrated to
afford (S)-2-mercapto-3-methylbutanoic acid as a white solid (2.00
g, 91%); .sup.1H NMR (300 MHz, CDCl.sub.3) .delta. 1.05 (d, J=6.7
Hz, 3H), 1.09 (d, J=6.7 Hz, 3H), 1.96 (d, J=9.7 Hz, 1H), 2.07 (m,
1H), 3.13 (dd, J=8.1, 9.7 Hz, 1H); .sup.13C NMR (75 MHz,
CDCl.sub.3) .delta. 19.3, 20.8, 32.6, 48.8, 179.8; IR (neat) 3100,
2966, 1705 cm.sup.-1; [.alpha.].sub.D.sup.22=-40.2 (c 1, ether).
##STR83##
[0258] (R)-Compound Y. To (R)-2-mercapto-3-methylbutanoic acid
(2.10 g, 15.6 mmol) was added aqueous NaOH (1.0 M in H.sub.2O, 37.6
mL, 37.6 mmol). The mixture was stirred at rt for 10 min. The
resulting solution was cooled to 0.degree. C., diluted with DMF (20
mL), and 4,6-dimethoxy-2-(methylsulfonyl)pyrimidine (3.42 g, 15.6
mmol) in DMF (10 mL) was added (a slightly modified procedure of
Fukuda, S.; Akiyoshi, Y.; Hori, K. J. Org. Chem. 1999, 64, 4768;
4,6-dimethoxy-2-(methylsulfonyl)pyrimidine is commercially
available from Aldrich). The mixture was warmed to rt and stirred
for 1 h. The resulting solution was quenched with 2 M HCl and
extracted with ethyl acetate. The combined extracts were washed
with brine, dried (Na.sub.2SO.sub.4) and concentrated. Purification
by silica gel chromatography (gradient column, ethyl
acetate-hexanes, 1:5.fwdarw.1:1.fwdarw.1:0) afforded (R)-compound Y
as white crystals (3.77 g, 88%); .sup.1H NMR (300 MHz, CDCl.sub.3)
.delta. 1.15 (d, J=6.4 Hz, 3H), 1.17 (J=6.4 Hz, 3H), 2.41 (m, 1H),
3.89 (s, 6H), 4.16 (d, J=5.9 Hz, 1H), 5.78 (s, 1H); .sup.13C NMR
(75 MHz, CDCl.sub.3) .delta. 20.1, 20.9, 30.1, 54.4, 54.9, 86.5,
169.4, 170.9, 177.9; IR (neat) 3050, 2964, 1710, 1581, 1557
cm.sup.-1; [.alpha.].sub.D.sup.22=+127.2 (c 1, CH.sub.2Cl.sub.2).
##STR84##
[0259] (S)-Compound Y. To (S)-2-mercapto-3-methylbutanoic acid
(2.00 g, 14.9 mmol) was added aqueous NaOH (1.0 M in H.sub.2O, 35.8
mL, 35.8 mmol). The mixture was stirred at rt for 10 min. The
resulting solution was cooled to .sup.0.degree. C., diluted with
DMF (20 mL), and 4,6-dimethoxy-2-(methylsulfonyl)pyrimidine.sup.2,3
(3.25 g, 14.9 mmol) in DMF (10 mL) was added. The mixture was
warmed to rt and stirred for 1 h. The resulting solution was
quenched with 2 M HCl and extracted with ethyl acetate. The
combined extracts were washed with brine, dried (Na.sub.2SO.sub.4)
and concentrated. Purification by silica gel chromatography
(gradient column, ethyl acetate-hexanes, 1:5.fwdarw.1:1.fwdarw.1:0)
afforded (S)-compound Y as white crystals (3.65 g, 90%); .sup.1H
NMR (300 MHz, CDCl.sub.3) .delta. 1.15 (d, J=6.4 Hz, 3H), 1.17
(J=6.4 Hz, 3H), 2.41 (m, 1H), 3.89 (s, 6H), 4.16 (d, J=5.9 Hz, 1H),
5.78 (s, 1H); .sup.13C NMR (75 MHz, CDCl.sub.3) .delta. 20.1, 20.9,
30.1, 54.4, 54.9, 86.5, 169.4, 170.9, 177.9; IR (neat) 3050, 2964,
1710, 1581, 1557 cm.sup.-1; [.alpha.].sub.D.sup.22=-127.3 (c 1,
CH.sub.2Cl.sub.2).
Example 7
Selective Activity of the S Enantiomer of Compound Y
[0260] Compound Y exists naturally in a racemic mixture, and has
the highly favorable PK properties described above. The experiments
described herein demonstrate that the S enantiomer of compound Y
stimulates erythroid cell proliferation significantly, while the R
enantiomer does not stimulate cell proliferation. The R enantiomer
of compound Y has activity in stimulating fetal globin expression
(see Table 3).
[0261] The proliferative activity of compound Y-S enantiomer was
tested in vitro in erythroid colonies cultured from peripheral
blood of 3 sources of individuals: sickle cell anemia patients who
were taking Hydroxyurea and have suppression of their endogenous
erythropoiesis, a normal adult subject, and two cord blood samples.
The erythroid cultures were established with low concentrations of
Erythropoietin (0.5 U/ml) alone and with one or the other
enantiomer of compound Y or the racemic mixture, or with high
concentrations of EPO (3 Units/ml, =3000 mU/ml, 100-fold the normal
physiologic concentration).
[0262] The cultures established in high EPO produced 25-45% (mean
35%) more colonies than the cultures established with the racemic
mix in low EPO or the R enantiomer in low EPO. Addition of the S--
enantiomer of compound Y at the same concentration resulted in a
mean of 45% more erythroid colonies than in the racemic mixture
(range 38-50%). Thus, the S enantiomer demonstrates the activity of
stimulating proliferation of erythroid colonies in vitro.
Example 8
Molecular Mechanism of Action
[0263] Without being bound by theory, it is thought that the
compounds described herein operate by a novel and highly specific
molecular mechanism of action as elucidated further below.
[0264] The erythroid kruppel-like factor, EKLF, is an essential
transcription factor for mammalian beta-like globin gene switch,
and it specifically activates transcription of the adult
beta-globin gene through binding of its zinc fingers to the
promoter. It has been shown that transcription factor EKLF is
required for activation of the gamma globin gene by the compounds
described herein. EKLF was previously considered to activate
primarily the beta (adult) globin gene. Transcription factor EKLF
is actively recruited to the gamma-globin gene promoter by the
compounds described herein. The human SWI/WNF complex is a
ubiquitous multimeric complex that regulates gene expression by
remodeling nucleosomal structure in an ATP-dependent manner. The
SWI/SNF complex contains one of two core ATPases, BRG1 or BRM.
These complexes can interact with sequence specific transcription
factors to either promote or repress target gene activation,
dependent on promoter context and complex content. The SWI/SNF
complex chromatin-modifying core ATPase Brg1 is required for gamma
globin induction by the compounds described herein. Brg1, the
co-activator SWI/SNF complex chromatin-modifying ATPase, is
actively recruited to the gamma-globin promoter by the compounds
described herein, and this recruitment is dependent upon the
presence of EKLF.
[0265] Two compounds were evaluated, compound P and W. Exposure of
primary erythroid cells to the high-potency inducer, compound P,
resulted in displacement of a repressor complex of HDAC3, NCoR,
specifically from the fetal globin gene promoter, with local
hyperacetylation of the promoter. Further, exposure to the compound
induced recruitment of erythroid kruppel-like factor (EKLF) and
Brahma-related gene 1 (Brg1) ATPase proteins to the gamma-globin
gene promoter, resulting in selective transcriptional activation of
the gamma globin gene. This is a selective effect on the fetal
(gamma globin) gene promoter, without generalized widespread
epistatic effects or any effects on the beta globin promoter, as
other agents tend to cause. The transcription factor EKLF and the
remodeling complex Brg1 and Pol II became bound to the promoter in
association with gene activation. In contrast, there were no
effects on the beta globin gene promoter. These compounds therefore
produce highly specific activating effects solely on the globin
gene promoter which would be beneficial to induce for therapy of
the beta globin gene disorders, and provide a targeted molecular
activity.
EQUIVALENTS
[0266] Those skilled in the art will recognize, or be able to
ascertain using no more than routine experimentation, numerous
equivalents to the specific procedures described herein. Such
equivalents are considered to be within the scope of the present
invention and are covered by the following claims. The contents of
all references, patents, and patent applications cited throughout
this application are hereby incorporated by reference. The
appropriate components, processes, and methods of those patents,
applications and other documents may be selected for the present
invention and embodiments thereof.
* * * * *