U.S. patent application number 12/761021 was filed with the patent office on 2010-08-05 for oxindoles as kinase inhibitors.
Invention is credited to David Bruge, Lars Thore BURGDORF, Hartmut Greiner, Maria Kordowicz, Christian Sirrenberg, Frank Zenke.
Application Number | 20100197757 12/761021 |
Document ID | / |
Family ID | 36977208 |
Filed Date | 2010-08-05 |
United States Patent
Application |
20100197757 |
Kind Code |
A1 |
BURGDORF; Lars Thore ; et
al. |
August 5, 2010 |
OXINDOLES AS KINASE INHIBITORS
Abstract
The present invention relates to oxindoles of the formula I,
their use as protein kinase activators or inhibitors, a method for
their manufacture, their use for the preparation of a medicament
for the treatment of diseases and their use for the manufacture of
a pharmaceutical composition. ##STR00001##
Inventors: |
BURGDORF; Lars Thore;
(Frankfurt, DE) ; Bruge; David; (Frankfurt,
DE) ; Greiner; Hartmut; (Weiterstadt, DE) ;
Kordowicz; Maria; (Griesheim, DE) ; Sirrenberg;
Christian; (Darmstadt, DE) ; Zenke; Frank;
(Darmstadt, DE) |
Correspondence
Address: |
MILLEN, WHITE, ZELANO & BRANIGAN, P.C.
2200 CLARENDON BLVD., SUITE 1400
ARLINGTON
VA
22201
US
|
Family ID: |
36977208 |
Appl. No.: |
12/761021 |
Filed: |
April 15, 2010 |
Related U.S. Patent Documents
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Application
Number |
Filing Date |
Patent Number |
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11916902 |
Dec 7, 2007 |
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PCT/EP2006/004423 |
May 11, 2006 |
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12761021 |
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Current U.S.
Class: |
514/418 |
Current CPC
Class: |
A61P 7/02 20180101; A61P
31/12 20180101; A61P 31/18 20180101; A61P 35/02 20180101; A61P
25/02 20180101; A61P 43/00 20180101; A61P 17/06 20180101; A61P
37/00 20180101; A61P 17/02 20180101; A61P 11/00 20180101; A61P 9/10
20180101; A61P 9/14 20180101; A61P 11/06 20180101; A61P 37/02
20180101; A61P 1/04 20180101; A61P 9/04 20180101; A61P 15/00
20180101; A61P 33/02 20180101; A61P 37/06 20180101; A61P 31/00
20180101; A61P 3/10 20180101; A61P 31/16 20180101; A61P 31/22
20180101; C07D 209/34 20130101; A61P 13/12 20180101; A61P 25/28
20180101; A61P 31/04 20180101; A61P 35/00 20180101; A61P 13/08
20180101; A61P 1/16 20180101; A61P 29/00 20180101; A61P 19/02
20180101; A61P 25/00 20180101; A61P 37/04 20180101; A61P 9/08
20180101; A61P 3/00 20180101; A61P 9/00 20180101; A61P 27/02
20180101; A61P 31/14 20180101 |
Class at
Publication: |
514/418 |
International
Class: |
A61K 31/404 20060101
A61K031/404; A61P 35/00 20060101 A61P035/00; A61P 35/02 20060101
A61P035/02 |
Foreign Application Data
Date |
Code |
Application Number |
Oct 6, 2005 |
EP |
05012559.0 |
Claims
1-28. (canceled)
29. A method for activating or inhibiting protein kinase is MEK1 or
MEK2, or for treating or preventing cancer, or for treating or
preventing melanoma, brain cancer, lung cancer, non-small cell lung
carcinoma, squamous cell cancer, colon cancer, duodenal cancer,
ductal cancer, colorectal cancer, gastric cancer, stomach cancer,
pancreatic cancer, hepatic cancer, renal cancer, bladder cancer,
endometrial cancer, ovarian cancer, uterine cancer, prostate
cancer, breast cancer, head cancer, neck cancer, oesophageal
cancer, gynecological cancer, dysplastic oral mucosa, polyposis,
invasive oral cancer, thyroid cancer, lymphoma, chronic leukaemia
or acute leukaemia, comprising administering to a subject in need
thereof an effective amount of a compound of formula I ##STR00033##
wherein X is (CH.sub.2).sub.p. R.sup.1 is Ar or Het, R.sup.2 is H,
A, Ar, (CH.sub.2).sub.mCON(R.sup.8).sub.2 or
(CH.sub.2).sub.mCONHAr, R.sup.3, R.sup.4, R.sup.6, and R.sup.7 are
independently from each other H, A, Ar, OR.sup.8, SR.sup.a, OAr,
SAr, N(R.sup.8).sub.2, NHAr, NAr.sub.2, Hal, NO.sub.2, CN,
COR.sup.S, COAr, NHCOA, NHCOAr, NHSO.sub.2A, NHSO.sub.2Ar,
SO.sub.2N(R.sup.8).sub.2, O(CH.sub.2).sub.nN(R.sup.8).sub.2 or
O(CH.sub.2).sub.nNHR.sup.8, R.sup.5 is H, A, Ar, OR.sup.8,
SR.sup.8, OAr, SAr, N(R.sup.8).sub.2, NHAr, NAr.sub.2, Hal,
NO.sub.2, CN, COAr, NHCOA, NHCOAr, NHSO.sub.2A, NHSO.sub.2Ar,
SO.sub.2N(R.sup.8).sub.2, O(CH.sub.2),N(R.sup.5).sub.2 or
O(CH.sub.2).sub.nNHR.sup.8, R.sup.8 is H, A or A-Ar, A is a linear
or branched alkyl or a cycloalkyl which is optionally substituted
by Hal, Ar is aryl, Het is heteroaryl, Hal Cl, Br, I or F, n, and p
are independently from each other 0-5, and m is 0-2, with the
provisio, that one of the residues R.sup.2, R.sup.3, R.sup.4,
R.sup.5, R.sup.6 or R.sup.7 is other than H and that
3-(1-Amino-2-phenyl-ethylidene)-1-methyl-1,3-dihydro-indol-2-one is
excluded, or a pharmaceutically acceptable salt, derivative,
prodrug, solvate or stereoisomer thereof, or a mixtures
thereof.
30. A method according to claim 29, wherein in the compound of
formula I X is (CH.sub.2).sub.p, p is 0-5, R.sup.1 is Ar or Het,
R.sup.2, R.sup.3, R.sup.4, R.sup.6, and R.sup.7 are H, and R.sup.5
is Hal.
31. A method according to claim 29, wherein in the compound of
formula I X is (CH.sub.2).sub.p, p is 0-5, R.sup.1 is Ar or Het,
R.sup.2, R.sup.3, R.sup.5, R.sup.6, and R.sup.7 are H, and R.sup.4
is Hal.
32. A method according to claim 29, wherein in the compound of
formula I X is (CH.sub.2).sub.p, p is 0-5, R.sup.1 is Ar or Het,
R.sup.3, R.sup.4, R.sup.5, R.sup.6, and R.sup.7 are H, and R.sup.2
is A or Ar.
33. A method according to claim 29, wherein in the compound of
formula I X is (CH.sub.2).sub.p, p is 0-5, R.sup.1 is Ar or Het,
R.sup.2, R.sup.3, R.sup.4, and R.sup.6 are H, R.sup.5 is Hal,
R.sup.7 is Hal or OR.sup.8, and R.sup.8 is H or A.
34. A method according to claim 29, wherein in the compound of
formula I X is (CH.sub.2).sub.p, p is 0-5, R.sup.1 is Ar or Het,
R.sup.2, R.sup.3, R.sup.5, and R.sup.6 are H, R.sup.4 is Hal,
R.sup.7 is Hal or OR.sup.8, and R.sup.8 is H or A.
35. A method according to claim 29, wherein in the compound of
formula I X is (CHO.sub.2).sub.p, p is 0-5, R.sup.1 is Ar or Het,
R.sup.2, R.sup.3, R.sup.4, and R.sup.6 are H, R.sup.5 is Hal,
R.sup.7 is Hal or OR.sup.8, and R.sup.8 is H or A.
36. A method according to claim 29, wherein in the compound of
formula I X is (CH.sub.2).sub.p, p is 0, R.sup.1 is phenyl,
R.sup.3, R.sup.4, R.sup.5, and R.sup.6 are H, R.sup.2 is A or Ar,
R.sup.7 is Hal or OR.sup.8, and R.sup.8 is H or A.
37. A method according to claim 29, wherein a compound of formula I
or a pharmaceutically acceptable salt thereof is administered.
38. A method according to claim 29, which is for activating or
inhibiting protein kinase is MEK1 or MEK2.
39. A method according to claim 29, which is for treating or
preventing cancer.
40. A method according to claim 29, which is for treating or
preventing melanoma, brain cancer, lung cancer, non-small cell lung
carcinoma, squamous cell cancer, colon cancer, duodenal cancer,
ductal cancer, colorectal cancer, gastric cancer, stomach cancer,
pancreatic cancer, hepatic cancer, renal cancer, bladder cancer,
endometrial cancer, ovarian cancer, uterine cancer, prostate
cancer, breast cancer, head cancer, neck cancer, oesophageal
cancer, gynecological cancer, dysplastic oral mucosa, polyposis,
invasive oral cancer, thyroid cancer, lymphoma, chronic leukaemia
or acute leukaemia.
41. A method according to claim 29, which is for treating melanoma,
brain cancer, lung cancer, non-small cell lung carcinoma, squamous
cell cancer, colon cancer, duodenal cancer, ductal cancer,
colorectal cancer, gastric cancer, stomach cancer, pancreatic
cancer, hepatic cancer, renal cancer, bladder cancer, endometrial
cancer, ovarian cancer, uterine cancer, prostate cancer, breast
cancer, head cancer, neck cancer, oesophageal cancer, gynecological
cancer, dysplastic oral mucosa, polyposis, invasive oral cancer,
thyroid cancer, lymphoma, chronic leukaemia or acute leukaemia.
42. A method according to claim 29, which further comprises
administering an estrogen receptor modulator, androgen receptor
modulator, retinoid receptor modulator, cytotoxic agent,
anti-proliferative agent, prenyl protein protease inhibitor, HMG
CoA reductase inhibitor, HIV protease inhibitor, reverse
transcriptase inhibitor, growth factor receptor inhibitor or
angiogenesis inhibitor.
43. A method according to claim 29, which further comprises
administering an anti-metastatic, antitumor or anti-angiogenic
agent, which is not a compound of formula I.
44. A method according to claim 29, which further comprises
providing radio therapy to the subject in need thereof.
45. A method for activating or inhibiting protein kinase is MEK1 or
MEK2, or for treating or preventing cancer, or for treating or
preventing melanoma, brain cancer, lung cancer, non-small cell lung
carcinoma, squamous cell cancer, colon cancer, duodenal cancer,
ductal cancer, colorectal cancer, gastric cancer, stomach cancer,
pancreatic cancer, hepatic cancer, renal cancer, bladder cancer,
endometrial cancer, ovarian cancer, uterine cancer, prostate
cancer, breast cancer, head cancer, neck cancer, oesophageal
cancer, gynecological cancer, dysplastic oral mucosa, polyposis,
invasive oral cancer, thyroid cancer, lymphoma, chronic leukaemia
or acute leukaemia, comprising administering to a subject in need
thereof an effective amount of a compound selected from the group
consisting of a)
3-(Amino-phenyl-methylene)-6-chloro-1,3-dihydro-indol-2-one, b)
3-(Amino-phenyl-methylene)-5-chloro-1,3-dihydro-indol-2-one, c)
3-[Amino-(4-hydroxy-phenyl)methylene]-1,3-dihydro-indol-2-one, d)
3-[Amino-(4-iodo-phenyl)methylene]-1,3-dihydro-indol-2-one, e)
3-(Amino-(4-iodo-phenyl)-methylene)-6-chloro-1,3-dihydro-indol-2-one,
f)
3-(Amino-(4-iodo-phenyl)-methylene)-5-chloro-1,3-dihydro-indol-2-one,
g) 3-[Amino-(3-iodo-phenyl)-methylene]-1,3-dihydro-indol-2-one, h)
3-(Amino-(3-iodo-phenyl)-methylene)-6-chloro-1,3-dihydro-indol-2-one,
i)
3-(Amino-(3-iodo-phenyl)-methylene)-5-chloro-1,3-dihydro-indol-2-one,
j)
3-(Amino-(3-iodo-phenyl)-methylene)-1-methyl-1,3-dihydro-indol-2-one,
k) 3-(Amino-phenyl-methylene)-5-bromo-1,3-dihydro-indol-2-one, l)
3-(Amino-(4-iodo-phenyl)-methylene)-1-methyl-1,3-dihydro-indol-2-one,
m)
3-(Amino-(4-iodo-phenyl)-methylene)-5-bromo-1,3-dihydro-indol-2-one,
n)
3-(Amino-(3-iodo-phenyl)-methylene)-5-bromo-1,3-dihydro-indol-2-one,
o) 3-[Amino-(4-methoxy-phenyl)-methylene]-1,3-dihydro-indol-2-one,
p)
3-(Amino-(4-methoxy-phenyl)-methylene)-6-chloro-1,3-dihydro-indol-2-one,
q)
3-(Amino-(4-methoxy-phenyl)-methylene)-5-bromo-1,3-dihydro-indol-2-one-
, r)
3-(Amino-(4-hydroxy-phenyl)-methylene)-6-chloro-1,3-dihydro-indol-2-o-
ne, s)
3-(Amino-(4-hydroxy-phenyl)-methylene)-5-chloro-1,3-dihydro-indol-2-
-one, t)
3-(Amino-phenyl-methylene)-1-methyl-1,3-dihydro-indol-2-one, u)
3-(Amino-(4-hydroxy-phenyl)-methylene)-1-methyl-1,3-dihydro-indol-2-one,
v)
3-(Amino-(4-hydroxy-phenyl)-methylene)-5-bromo-1,3-dihydro-indol-2-one-
, w)
3-(Amino-(4-methoxy-phenyl)-methylene)-5-chloro-1,3-dihydro-indol-2-o-
ne, x)
3-[Amino-(4-fluoro-phenyl)methylene]-1,3-dihydro-indol-2-one, y)
3-(Amino-(4-fluoro-phenyl)-methylene)-5-chloro-1,3-dihydro-indol-2-one,
and z)
3-(Amino-(4-methoxy-phenyl)-methylene)-1-methyl-1,3-dihydro-indol--
2-one, and the physiologically acceptable salts, derivatives,
prodrugs, solvates and stereoisomers thereof, including mixtures
thereof in all ratios.
46. A method according to claim 45, wherein a)
3-(Amino-phenyl-methylene)-6-chloro-1,3-dihydro-indol-2-one, b)
3-(Amino-phenyl-methylene)-5-chloro-1,3-dihydro-indol-2-one, c)
3-[Amino-(4-hydroxy-phenyl)-methylene]-1,3-dihydro-indol-2-one, d)
3-[Amino-(4-iodo-phenyl)-methylene]-1,3-dihydro-indol-2-one, e)
3-(Amino-(4-iodo-phenyl)-methylene)-6-chloro-1,3-dihydro-indol-2-one,
f)
3-(Amino-(4-iodo-phenyl)-methylene)-5-chloro-1,3-dihydro-indol-2-one,
g) 3-[Amino-(3-iodo-phenyl)-methylene]-1,3-dihydro-indol-2-one, h)
3-(Amino-(3-iodo-phenyl)-methylene)-6-chloro-1,3-dihydro-indol-2-one,
i)
3-(Amino-(3-iodo-phenyl)-methylene)-5-chloro-1,3-dihydro-indol-2-one,
j)
3-(Amino-(3-iodo-phenyl)-methylene)-1-methyl-1,3-dihydro-indol-2-one,
k) 3-(Amino-phenyl-methylene)-5-bromo-1,3-dihydro-indol-2-one, l)
3-(Amino-(4-iodo-phenyl)-methylene)-1-methyl-1,3-dihydro-indol-2-one,
m)
3-(Amino-(4-iodo-phenyl)-methylene)-5-bromo-1,3-dihydro-indol-2-one,
n)
3-(Amino-(3-iodo-phenyl)-methylene)-5-bromo-1,3-dihydro-indol-2-one,
o) 3-(Amino-(4-methoxy-phenyl)-methylene]-1,3-dihydro-indol-2-one,
p)
3-(Amino-(4-methoxy-phenyl)-methylene)-6-chloro-1,3-dihydro-indol-2-one,
q)
3-(Amino-(4-methoxy-phenyl)methylene)-5-bromo-1,3-dihydro-indol-2-one,
r)
3-(Amino-(4-hydroxy-phenyl)-methylene)-6-chloro-1,3-dihydro-indol-2-on-
e, s)
3-(Amino-(4-hydroxy-phenyl)methylene)-5-chloro-1,3-dihydro-indol-2-o-
ne, t) 3-(Amino-phenyl-methylene)-1-methyl-1,3-dihydro-indol-2-one,
u)
3-(Amino-(4-hydroxy-phenyl)-methylene)-1-methyl-1,3-dihydro-indol-2-one,
v)
3-(Amino-(4-hydroxy-phenyl)-methylene)-5-bromo-1,3-dihydro-indol-2-one-
, w)
3-(Amino-(4-methoxy-phenyl)-methylene)-5-chloro-1,3-dihydro-indol-2-o-
ne, x)
3-(Amino-(4-fluoro-phenyl)-methylene]-1,3-dihydro-indol-2-one, y)
3-(Amino-(4-fluoro-phenyl)-methylene)-5-chloro-1,3-dihydro-indol-2-one,
or z)
3-(Amino-(4-methoxy-phenyl)-methylene)-1-methyl-1,3-dihydro-indol-2-
-one, or a physiologically acceptable salt thereof is
administered.
47. A method according to claim 45, which is for treating melanoma,
brain cancer, lung cancer, non-small cell lung carcinoma, squamous
cell cancer, colon cancer, duodenal cancer, ductal cancer,
colorectal cancer, gastric cancer, stomach cancer, pancreatic
cancer, hepatic cancer, renal cancer, bladder cancer, endometrial
cancer, ovarian cancer, uterine cancer, prostate cancer, breast
cancer, head cancer, neck cancer, oesophageal cancer, gynecological
cancer, dysplastic oral mucosa, polyposis, invasive oral cancer,
thyroid cancer, lymphoma, chronic leukaemia or acute leukaemia.
Description
FIELD OF THE INVENTION
[0001] The present invention relates to oxindoles, their use as
protein kinase activators or inhibitors, a method for their
manufacture, their use for the preparation of a medicament for the
treatment of diseases and their use for the manufacture of a
pharmaceutical composition.
BACKGROUND OF THE INVENTION
[0002] Protein kinases are involved in signaling pathways for such
important cellular activities as responses to extracellular signals
and cell cycle checkpoints. Inhibition or activation of specific
protein kinases provides a means of intervening in these signaling
pathways, for example to block the effect of an extracellular
signal, to release a cell from cell cycle checkpoint, etc. Defects
in the activity of protein kinases are associated with a variety of
pathological or clinical conditions, where there is a defect in the
signaling mediated by protein kinases. Such conditions include
those associated with defects in cell cycle regulation or in
response to extracellular signals, e.g., immunological disorders,
autoimmune and immunodeficiency diseases; hyperproliferative
disorders, which may include psoriasis, arthritis, inflammation,
endometriosis, scarring, cancer, etc.
[0003] Aberrant activation of the Ras-Raf-MAP kinase signaling
pathway contributes to the process of tumorigenesis, i.e. the
conversion of a normal into a malignant cell. Many molecular
details have been worked out, how the MAPK signaling module
initiates proliferation or inhibits apoptotic response, thus
explaining how the homeostatic balance between proliferation and
cell death can be disturbed. MKK1 and MKK2, also known as MEK1 and
MEK2, represent a key element in this signaling cascade. MEKs are
activated by Raf kinase phosphorylation and, in turn, phosphorylate
and activate their bona fide substrates ERK1 and ERK2. Oncogenic
Ras mutations, mutations in the effector kinase B-Raf and even
growth factor overexpression and mutation may lead to persistent
activation of the MAPK pathway justifying the potential of MAPK
pathway inhibitors at a level of Rat, MEK or ERK as promising
anticancer therapeutics.
[0004] Therefore, compounds, which are active in modulating
purified kinase proteins, e.g. there is a modulation in the
phosphorylation of a specific substrate in the presence of the
compound, can be used for the treatment of protein kinase-dependent
diseases and conditions, such as cancer, tumour growth,
artherosclerosis, age-related macular degeneration, diabetic
retinopathy, inflammatory diseases and the like, in mammals.
[0005] Amino-oxindoles are known from e.g. WO9952869, WO9962882,
WO04026829, WO04009546, WO04009547, EP104860, U.S. Pat. No.
4,145,422, WO03027102 and WO0149287.
3-(1-Amino-2-phenyl-ethylidene)-1-methyl-1,3-dihydro-indol-2-one is
known from Wenkert, E. et al., J. Am. Chem. Soc. (1958), 80,
4899-4903, 3-(Amino-phenyl-methylene)-1,3-dihydro-indol-2-one is
known from Stauss, U. et al., Helv. Chim. Acta (1972), 55(3),
771-780.
[0006] Compounds described as putative MEK inhibitors are known
from e.g. WO03062191, WO04056789, WO03077914, WO03077855,
WO04041811 and WO04048386.
[0007] Thus, as there remains a need in advantageous therapeutics,
a preferred object of the present invention was to provide new
pharmaceutically active compounds. A particularly preferred aim of
the present invention was to provide effective modulators of one or
more protein kinases, especially selected from the group of Raf,
MEK, PKB, Tie2, PDGFR, Met, SGK1, IGF1R and VEGFR.
BRIEF DESCRIPTION OF THE INVENTION
[0008] Surprisingly, the compounds of the general formula I show
pharmaceutical activities as they act as effective modulators
(activators or inhibitors) of one or more protein kinases selected
from the group of Raf, MEK, PKB, Tie2, PDGFR, Met, SGK1, IGF1R and
VEGFR.
[0009] Therefore, an embodiment of the present invention are
compounds of the formula I,
##STR00002##
wherein [0010] X is (CH.sub.2).sub.p, [0011] R.sup.1 is Ar or Het,
[0012] R.sup.2 is H, A, Ar, (CH.sub.2).sub.mCON(R.sup.8).sub.2,
(CH.sub.2).sub.mCONHAr, S(O).sub.mA, S(O).sub.mAr, NHCOA, NHCOAr,
NHSO.sub.2A, NHSO.sub.2Ar, SO.sub.2N(R.sup.8).sub.2,
N(CH.sub.2).sub.nC(CH.sub.3).sub.2(CH.sub.2).sub.nN(R.sup.8).sub.2,
(CH.sub.2).sub.nN(R.sup.8)SO.sub.mA,
(CH.sub.2).sub.nN(R.sup.8)SO.sub.mAr, (CH.sub.2).sub.nSO.sub.mA,
(CH.sub.2).sub.nSO.sub.mAr or (CH.sub.2).sub.nSO.sub.mN(R.sup.8)A,
[0013] R.sup.3, R.sup.4, R.sup.5, R.sup.6, R.sup.7 are
independently from each other H, A, Ar, OR.sup.B, SR.sup.8, OAr,
SAr, N(R.sup.8).sub.2, NHAr, NAr.sub.2, Hal, NO.sub.2, CN, COW,
COAr, S(O).sub.mA, S(O).sub.mAr, NHCOA, NHCOAr, NHSO.sub.2A,
NHSO.sub.2Ar, SO.sub.2N(R.sup.8).sub.2,
O(CH.sub.2).sub.nN(R.sup.8).sub.2, O(CH.sub.2).sub.nNHR.sup.8,
O(CH.sub.2).sub.n-morpholine, O(CH.sub.2).sub.n-piperazine,
O(CH.sub.2).sub.n-pyrrolidine, O(CH.sub.2).sub.n-piperidine,
O-piperidine, O(CH.sub.2).sub.n-oxopiperazine,
O(CH.sub.2).sub.n-oxomorpholine, O(CH.sub.2).sub.n-oxopyrrolidine,
O(CH.sub.2).sub.nC(CH.sub.3).sub.2(CH.sub.2).sub.nN(R.sup.8).sub.2,
N(CH.sub.2).sub.nC(CH.sub.3).sub.2(CH.sub.2).sub.nN(R.sup.8).sub.2,
O(CH.sub.2).sub.nN(R.sup.8)SO.sub.mA,
O(CH.sub.2).sub.nN(R.sup.8)SO.sub.mAr,
O(CH.sub.2).sub.nN(R.sup.8)SO.sub.mN(R.sup.8).sub.2A,
(CH.sub.2).sub.nN(R.sup.8)SO.sub.mA,
(CH.sub.2).sub.nN(R.sup.8)SO.sub.mAr,
(CH.sub.2).sub.nN(R.sup.8)SO.sub.mN(R.sup.8).sub.2A,
O(CH.sub.2).sub.nSO.sub.mA, O(CH.sub.2).sub.nSO.sub.mAr,
O(CH.sub.2).sub.nSO.sub.mN(R.sup.8)A, (CH.sub.2).sub.nSO.sub.mA,
(CH.sub.2).sub.nSO.sub.mAr or (CH.sub.2).sub.nSO.sub.mN(R.sup.8)A,
[0014] R.sup.8 is H, A or A-Ar, [0015] A is a linear or branched
alkyl or a cycloalkyl which is optionally substituted by Hal,
[0016] Ar is aryl, [0017] Het is heteroaryl, [0018] Hal Cl, Br, I
or F, [0019] n, p are independently from each other 0-5, [0020] m
is 0-2, with the provisio, that one of the residues R.sup.2,
R.sup.3, R.sup.4, R.sup.5, R.sup.6 or R.sup.7 is other than H and
that
3-(1-Amino-2-phenyl-ethylidene)-1-methyl-1,3-dihydro-indol-2-one is
excluded, and the physiologically acceptable salts, derivatives,
prodrugs, solvates and stereoisomers thereof, including mixtures
thereof in all ratios.
[0021] A further preferred embodiment of the present invention are
compounds according to formula I, wherein [0022] X is
(CH.sub.2).sub.p, [0023] R.sup.1 is Ar or Het, [0024] R.sup.2 is H,
A, Ar, (CH.sub.2).sub.mCON(R.sup.8).sub.2, (CH.sub.2).sub.mCONHAr,
(CH.sub.2).sub.nN(R.sup.8)SO.sub.mA,
(CH.sub.2).sub.nN(R.sup.8)SO.sub.mAr or
(CH.sub.2).sub.nSO.sub.mN(R.sup.8)A, [0025] R.sup.3, R.sup.4,
R.sup.5, R.sup.6, R.sup.7 are independently from each other H, A,
Ar, OR.sup.8, SR.sup.8, OAr, SAr, N(R.sup.8).sub.2, NHAr,
NAr.sub.2, Hal, NO.sub.2, ON, COR.sup.S, COAr, NHCOA, NHCOAr,
NHSO.sub.2A, NHSO.sub.2Ar, SO.sub.2N(R.sup.8).sub.2,
O(CH.sub.2).sub.nN(R.sup.8).sub.2, O(CH.sub.2).sub.nNHR.sup.8,
O(CH.sub.2).sub.n-morpholine, O(CH.sub.2).sub.n-piperazine,
O(CH.sub.2).sub.n-pyrrolidine, O(CH.sub.2).sub.n-piperidine,
O-piperidine, O(CH.sub.2).sub.n-oxopiperazine,
O(CH.sub.2).sub.n-oxomorpholine, O(CH.sub.2).sub.n-oxopyrrolidine,
O(CH.sub.2).sub.nC(CH.sub.3).sub.2(CH.sub.2).sub.nN(R.sup.8).sub.2
or
N(CH.sub.2).sub.nC(CH.sub.3).sub.2(CH.sub.2).sub.nN(R.sup.8).sub.2,
[0026] R.sup.8 is H, A or A-Ar, [0027] A is a linear or branched
alkyl or a cycloalkyl which is optionally substituted by Hal,
[0028] Ar is aryl, [0029] Het is heteroaryl, [0030] Hal Cl, Br, I
or F, [0031] n, p are independently from each other 0-5, [0032] m
is 0-2, with the provisio, that one of the residues R.sup.2,
R.sup.3, R.sup.4, R.sup.5, R.sup.6 or R.sup.7 is other than H and
that
3-(1-Amino-2-phenyl-ethylidene)-1-methyl-1,3-dihydro-indol-2-one is
excluded, and the physiologically acceptable salts, derivatives,
prodrugs, solvates and stereoisomers thereof, including mixtures
thereof in all ratios.
[0033] A further preferred embodiment of the present invention are
compounds according to formula I, wherein [0034] X is
(CH.sub.2).sub.p, [0035] R.sup.1 is Ar or Het, [0036] R.sup.2 is H,
A, Ar, (CH.sub.2).sub.mCON(R.sup.8).sub.2 or
(CH.sub.2).sub.mCONHAr, [0037] R.sup.3, R.sup.4, R.sup.5, R.sup.6,
R.sup.7 are independently from each other H, A, Ar, OR.sup.8,
SR.sup.8, OAr, SAr, N(R.sup.8).sub.2, NHAr, NAr.sub.2, Hal,
NO.sub.2, CN, COR.sup.8, COAr, NHCOA, NHCOAr, NHSO.sub.2A,
NHSO.sub.2Ar, SO.sub.2N(R.sup.8).sub.2,
O(CH.sub.2).sub.nN(R.sup.8).sub.2 or O(CH.sub.2).sub.nNHR.sup.8,
[0038] R.sup.8 is H, A or A-Ar, [0039] A is a linear or branched
alkyl or a cycloalkyl which is optionally substituted by Hal,
[0040] Ar is aryl, [0041] Het is heteroaryl, [0042] Hal Cl, Br, I
or F, [0043] n, p are independently from each other 0-5, [0044] m
is 0-2, with the provisio, that one of the residues R.sup.2,
R.sup.3, R.sup.4, R.sup.5, R.sup.6 or R.sup.7 is other than H and
that
3-(1-Amino-2-phenyl-ethylidene)-1-methyl-1,3-dihydro-indol-2-one is
excluded, and the physiologically acceptable salts, derivatives,
prodrugs, solvates and stereoisomers thereof, including mixtures
thereof in all ratios.
[0045] A further preferred embodiment of the present invention are
compounds according to formula I, wherein [0046] X is
(CH.sub.2).sub.p, [0047] p is 0-5, [0048] R.sup.1 is Ar or Het,
[0049] R.sup.2, R.sup.3, R.sup.4, R.sup.6, R.sup.7 are H, [0050]
R.sup.5 is Hal, and the physiologically acceptable salts,
derivatives, prodrugs, solvates and stereoisomers thereof,
including mixtures thereof in all ratios.
[0051] A further preferred embodiment of the present invention are
compounds according to formula I, wherein [0052] X is
(CH.sub.2).sub.p, [0053] p is 0-5, [0054] R.sup.1 is Ar or Het,
[0055] R.sup.2, R.sup.3, R.sup.4, R.sup.6, R.sup.7 are H, [0056]
R.sup.4 is Hal, and the physiologically acceptable salts,
derivatives, prodrugs, solvates and stereoisomers thereof,
including mixtures thereof in all ratios.
[0057] A further preferred embodiment of the present invention are
compounds according to formula I, wherein [0058] X is
(CH.sub.2).sub.p, [0059] p is 0-5, [0060] R.sup.1 is Ar or Het,
[0061] R.sup.2, R.sup.3, R.sup.4, R.sup.6, R.sup.7 are H, [0062]
R.sup.2, is A or Ar, with the provisio, that
3-(1-Amino-2-phenyl-ethylidene)-1-methyl-1,3-dihydroindol-2-one is
excluded, and the physiologically acceptable salts, derivatives,
prodrugs, solvates and stereoisomers thereof, including mixtures
thereof in all ratios.
[0063] A further preferred embodiment of the present invention are
compounds according to formula I, wherein [0064] X is
(CH.sub.2).sub.p, [0065] p is 0-5, [0066] R.sup.1 is Ar or Het,
[0067] R.sup.2, R.sup.3, R.sup.4, R.sup.6 are H, [0068] R.sup.5 is
Hal, [0069] R.sup.7 is Hal or OR.sup.8, [0070] R.sup.8 is H or A,
and the physiologically acceptable salts, derivatives, prodrugs,
solvates and stereoisomers thereof, including mixtures thereof in
all ratios.
[0071] A further preferred embodiment of the present invention are
compounds according to formula I, wherein [0072] X is
(CH.sub.2).sub.p, [0073] p is 0-5, [0074] R.sup.1 is Ar or Het,
[0075] R.sup.2, R.sup.3, R.sup.5, R.sup.6 are H, [0076] R.sup.4 is
Hal, [0077] R.sup.7 is Hal or OR.sup.8, [0078] R.sup.8 is H or A,
and the physiologically acceptable salts, derivatives, prodrugs,
solvates and stereoisomers thereof, including mixtures thereof in
all ratios.
[0079] A further preferred embodiment of the present invention are
compounds according to formula I, wherein [0080] X is
(CH.sub.2).sub.p, [0081] p is 0-5, [0082] R.sup.1 is Ar or Het,
[0083] R.sup.2, R.sup.3, R.sup.4, R.sup.6 are H, [0084] R.sup.5 is
Hal, [0085] R.sup.7 is Hal or OR.sup.8, [0086] R.sup.8 is H or A,
and the physiologically acceptable salts, derivatives, prodrugs,
solvates and stereoisomers thereof, including mixtures thereof in
all ratios.
[0087] A further preferred embodiment of the present invention are
compounds according to formula I, wherein [0088] X is
(CH.sub.2).sub.p, [0089] p is 0, [0090] R.sup.1 is phenyl, [0091]
R.sup.2, R.sup.3, R.sup.4, R.sup.6 are H, [0092] R.sup.2 is A or
Ar, [0093] R.sup.7 is Hal or OR.sup.8, [0094] R.sup.8 is H or A,
and the physiologically acceptable salts, derivatives, prodrugs,
solvates and stereoisomers thereof, including mixtures thereof in
all ratios.
[0095] An especially preferred embodiment of the present invention
are compounds according to formula I, selected from the group
consisting of [0096] a)
3-(Amino-phenyl-methylene)-6-chloro-1,3-dihydro-indol-2-one, [0097]
b) 3-(Amino-phenyl-methylene)-5-chloro-1,3-dihydro-indol-2-one,
[0098] c)
3-[Amino-(4-hydroxy-phenyl)-methylene]-1,3-dihydro-indol-2-one
[0099] d)
3-[Amino-(4-iodo-phenyl)-methylene]-1,3-dihydro-indol-2-one [0100]
e) 3-(Amino-(4-iodo-phenyl)-methylene)-6-chloro-1,3-dihydro-indol--
2-one, [0101] f)
3-(Amino-(4-iodo-phenyl)-methylene)-5-chloro-1,3-dihydro-indol-2-one,
[0102] g)
3-[Amino-(3-iodo-phenyl)-methylene]-1,3-dihydro-indol-2-one [0103]
h) 3-(Amino-(3-iodo-phenyl)-methylene)-6-chloro-1,3-dihydro-indol--
2-one, [0104] i)
3-(Amino-(3-iodo-phenyl)-methylene)-5-chloro-1,3-dihydro-indol-2-one,
[0105] j)
3-(Amino-(3-iodo-phenyl)-methylene)-1-methyl-1,3-dihydro-indol--
2-one, [0106] k)
3-(Amino-phenyl-methylene)-5-bromo-1,3-dihydro-indol-2-one,
[0107] l)
3-(Amino-(4-iodo-phenyl)-methylene)-1-methyl-1,3-dihydro-indol-2-
-one, [0108] m)
3-(Amino-(4-iodo-phenyl)-methylene)-5-bromo-1,3-dihydro-indol-2-one
[0109] n)
3-(Amino-(3-iodo-phenyl)-methylene)-5-bromo-1,3-dihydro-indol-2-
-one, [0110] o)
3-[Amino-(4-methoxy-phenyl)methylene]-1,3-dihydro-indol-2-one
[0111] p)
3-(Amino-(4-methoxy-phenyl)-methylene)-6-chloro-1,3-dihydro-indol-2-one,
[0112] q)
3-(Amino-(4-methoxy-phenyl)-methylene)-5-bromo-1,3-dihydro-indo-
l-2-one, [0113] r)
3-(Amino-(4-hydroxy-phenyl)-methylene)-6-chloro-1,3-dihydro-indol-2-one,
[0114] s)
3-(Amino-(4-hydroxy-phenyl)-methylene)-5-chloro-1,3-dihydro-ind-
ol-2-one, [0115] t)
3-(Amino-phenyl-methylene)-1-methyl-1,3-dihydro-indol-2-one, [0116]
u)
3-(Amino-(4-hydroxy-phenyl)-methylene)-1-methyl-1,3-dihydro-indol-2-one,
[0117] v)
3-(Amino-(4-hydroxy-phenyl)-methylene)-5-bromo-1,3-dihydro-indo-
l-2-one, [0118] w)
3-(Amino-(4-methoxy-phenyl)-methylene)-5-chloro-1,3-dihydro-indol-2-one,
[0119] x)
3-[Amino-(4-fluoro-phenyl)-methylene]-1,3-dihydro-indol-2-one
[0120] y)
3-(Amino-(4-fluoro-phenyl)-methylene)-5-chloro-1,3-dihydro-indo-
l-2-one, [0121] z)
3-(Amino-(4-methoxy-phenyl)-methylene)-1-methyl-1,3-dihydro-indol-2-one,
and the physiologically acceptable salts, derivatives, prodrugs,
solvates and stereoisomers thereof, including mixtures thereof in
all ratios.
[0122] Suitable salts and pharmaceutically acceptable salts of the
compounds according to the invention are conventional non-toxic
salts and include acid addition salts such as organic acid salts
(e.g. acetate, trifluoroacetate, maleate, tartrate,
methanesulfonate, benzenesulfonate, formate, toluenesulfonate),
inorganic acid salt (e.g. hydrochloride, hydrobromide, hydroiodide,
sulfate, nitrate, phosphate), or salts with an amino acid (e.g.
arginine, aspartic acid, glutamic acid), or metal salts such as
alkali metal salts (e.g. sodium salt, potassium salt) and alkaline
earth metal salts (e.g. calcium salt, magnesium salt), ammonium
salts, or organic base salts (e.g. trimethylamine salt,
triethylamine salt, pyridine salt, picoline salt, dicyclohexylamine
salt, N,N'-dibenzylethylenediamine salt).
[0123] The term "pharmaceutically usable derivatives" or
"pharmaceutically acceptable derivatives" is taken to mean, for
example, the salts of the compounds according to the invention and
so-called prodrug compounds. The term refers to any
pharmaceutically acceptable derivative of a compound of the present
invention, for example, an ester or an amide, which upon
administration to a mammal is capable of providing (directly or
indirectly) a compound of the present invention or an active
metabolite thereof. Such derivatives are clear to those skilled in
the art, without undue experimentation, and with reference to the
teaching of Burger's Medicinal Chemistry And Drug Discovery, 5th
Edition, Vol 1: Principles and Practice, which is incorporated
herein by reference to the extent that it teaches physiologically
functional derivatives.
[0124] The term "prodrug derivatives" is taken to mean, for
example, compounds of the present invention which have been
modified, for example, with alkyl or acyl groups, sugars or
oligopeptides and which are rapidly cleaved in the organism and
thus release the active ingredients according to the invention.
These also include biodegradable polymer derivatives of the
compounds according to the invention, as described, for example, in
Int. J. Pharm. 115, 61-67 (1995).
[0125] The invention also relates to mixtures of the compounds
according to the invention, for example mixtures of two
diastereomers, for example in the ratio 1:1, 1.2, 1:3, 1:4, 1:5,
1:10, 1:100 or 1:1000. These are particularly preferably mixtures
of stereoisomeric compounds.
[0126] Additionally, the invention comprises the polymorphic forms
of the compounds according to the invention, e.g. the amorphic and
crystalline polymorphic forms.
[0127] As used herein, the term "solvate" preferably refers to a
complex of variable stoichiometry formed by a solute and a solvent.
The term solvates of the compounds is therefore taken to mean
adductions of inert solvent molecules onto the compounds, which
form owing to their mutual attractive force. Such solvents for the
purpose of the invention may not interfere with the biological
activity of the solute. Preferably the solvent used is a
pharmaceutically acceptable solvent. Examples of suitable
pharmaceutically acceptable solvents include, without limitation,
water, ethanol and acetic acid. Most preferably the solvent used is
water. Solvates are, for example, monohydrates, dihydrates or
alcoholates.
[0128] Certain of the compounds described herein may contain one or
more chiral atoms, or may otherwise be capable of existing as two
or more stereoisomers, which are usually enantiomers and/or
diastereomers. Accordingly, the compounds of this invention include
mixtures of stereoisomers, especially mixtures of enantiomers, as
well as purified stereoisomers, especially purified enantiomers, or
stereoisomerically enriched mixtures, especially enantiomerically
enriched mixtures. Also included within the scope of the invention
are the individual isomers of the compounds of the present
invention as well as any wholly or partially equilibrated mixtures
thereof. The present invention also covers the individual isomers
of the compounds represented by the formulas above as mixtures with
isomers thereof in which one or more chiral centers are inverted.
Also, it is understood that all tautomers and mixtures of tautomers
of the compounds of the present invention are included within the
scope of the compounds of the present invention and preferably the
formulae and subformulae corresponding thereto.
[0129] Racemates obtained can be resolved into the isomers
mechanically or chemically by methods known per se. Diastereomers
are preferably formed from the racemic mixture by reaction with an
optically active resolving agent. Examples of suitable resolving
agents are optically active acids, such as the D and L forms of
tartaric acid, diacetyltartaric acid, dibenzoyltartaric acid,
mandelic acid, malic acid, lactic acid or the various optically
active camphorsulfonic acids, such asp-camphorsulfonic acid. Also
advantageous is enantiomer resolution with the aid of a column
filled with an optically active resolving agent (for example
dinitrobenzoylphenylglycine); an example of a suitable eluent is a
hexane/isopropanol/acetonitrile mixture.
[0130] The diastereomer resolution can also be carried out by
standard purification processes, such as, for example,
chromatography or fractional crystallization.
[0131] It is of course also possible to obtain optically active
compounds of the present invention by the methods described above
by using starting materials which are already optically active.
[0132] Unless indicated otherwise, it is to be understood that
reference to the compounds of the present invention preferably
includes the reference to the subformulae corresponding thereto. It
is also understood that the following embodiments, including uses
and compositions, although recited with respect to the compounds of
the present invention are preferably also applicable to
subformulae.
[0133] As used herein, the terms "group", "residue" and "radical"
or "groups", "residues" and "radicals" are usually used as
synonyms, respectively, as it is common practice in the art.
[0134] As used herein, the term "optionally" means that the
subsequently described event(s) may or may not occur, and includes
both event(s), which occur, and events that do not occur.
[0135] As used herein, the term "substituted" preferably refers to
substitution with the named substituent or substituents, multiple
degrees of substitution being allowed unless otherwise stated.
[0136] Subject of the present invention are especially compounds of
the present invention in which one or more substituents or groups,
preferably the major part of the substituents or groups has a
meaning which is indicated as preferred, more preferred, even more
preferred or especially preferred.
[0137] As used herein, the term "halogen" or "hal" preferably
refers to fluorine (F), chlorine (Cl), bromine (Br) or iodine
(I).
[0138] As used herein, the term "A" or "alkyl" preferably refers to
a straight or branched chain hydrocarbon having from one to twelve
carbon atoms, wherein optionally 1-5H atoms are replaced by F
and/or Cl, multiple degrees of substitution being allowed. Examples
of "alkyl" as used herein include, but are not limited to, methyl,
ethyl, n-propyl, isopropyl, n-butyl, isobutyl, t-butyl, n-pentyl,
isopentyl, and the like.
[0139] As used herein, the term "cycloalkyl" preferably refers to a
non-aromatic cyclic hydrocarbon ring system, with one or more rings
attached to each other, each ring preferably having from three to
seven carbon atoms, which optionally includes an alkyl linker,
preferably a C.sub.1-C.sub.6 alkyl linker, through which it may be
attached. Optionally, in the "cycloalkyl" 1-5H atoms are replaced
by F and/or Cl, multiple degrees of substitution being allowed. The
alkyl or C.sub.1-C.sub.6 alkyl group is as defined above. Exemplary
"cycloalkyl" groups include, but are not limited to, cyclopropyl,
cyclobutyl, cyclopentyl, cyclohexyl and cycloheptyl.
[0140] As used herein, the term "Ar" or "aryl" preferably refers to
a benzene ring or a benzene ring system, which is optionally
substituted by A, Ar, OR.sup.8, SR.sup.8, OAr, SAr,
N(R.sup.8).sub.2, NHAr, NAr.sub.2, Hal, NO.sub.2, ON, COR.sup.8,
COAr, S(O).sub.mA, S(O).sub.mAr, NHCOA, NHCOAr, NHSO.sub.2A,
NHSO.sub.2Ar, SO.sub.2N(R.sup.8).sub.2,
O(CH.sub.2).sub.nN(R.sup.8).sub.2, O(CH.sub.2).sub.nNHR.sup.8,
O(CH.sub.2).sub.n-morpholine, O(CH.sub.2).sub.n-piperazine,
O(CH.sub.2).sub.n-pyrrolidine, O(CH.sub.2).sub.n-piperidine,
O-piperidine, O(CH.sub.2).sub.n-oxopiperazine,
O(CH.sub.2).sub.n-oxomorpholine, O(CH.sub.2).sub.n-oxopyrrolidine,
O(CH.sub.2).sub.nC(CH.sub.3).sub.2(CH.sub.2).sub.nN(R.sup.8).sub.2,
N(CH.sub.2).sub.nC(CH.sub.3).sub.2(CH.sub.2).sub.nN(R.sup.8).sub.2,
O(CH.sub.2).sub.nN(R.sup.8)SO.sub.mA,
O(CH.sub.2).sub.nN(R.sup.8)SO.sub.mAr,
O(CH.sub.2).sub.nN(R.sup.8)SO.sub.mN(R.sup.8).sub.2A,
(CH.sub.2).sub.nN(R.sup.8)SO.sub.mA,
(CH.sub.2).sub.nN(R.sup.8)SO.sub.mAr,
(CH.sub.2).sub.nN(R.sup.8)SO.sub.mN(R.sup.8).sub.2A,
O(CH.sub.2).sub.nSO.sub.mA, O(CH.sub.2).sub.nSO.sub.mAr,
O(CH.sub.2).sub.nSO.sub.mN(R.sup.8)A, (CH.sub.2).sub.nSO.sub.mA,
(CH.sub.2).sub.nSO.sub.mAr, (CH.sub.2).sub.nSO.sub.mN(R.sup.8)A,
and the like, multiple degrees of substitution being allowed.
Examples of "aryl" groups include, but are not limited to Phenyl,
2-naphthyl, 1-naphthyl, biphenyl, anthracyl, phenanthracyl, as well
as substituted derivatives thereof.
[0141] As used herein, the term "Het" or the term "heteroaryl"
preferably refers to a monocyclic five to seven-membered aromatic
ring, or to a fused bicyclic aromatic ring system comprising two of
such monocyclic five to seven-membered aromatic rings. These
heteroaryl rings contain one or more nitrogen, sulfur and/or oxygen
heteroatoms, where N-oxides and sulfur oxides and dioxides are
permissible heteroatom substitutions and may be optionally
substituted by one or more substituents, selected from the group
consisting of A, Ar, OR.sup.8, SR.sup.8, OAr, SAr,
N(R.sup.8).sub.2, NHAr, NAr.sub.2, Hal, NO.sub.2, ON, COR.sup.8,
COAr, S(O).sub.mA, S(O).sub.mAr, NHCOA, NHCOAr, NHSO.sub.2A,
NHSO.sub.2Ar, SO.sub.2N(R.sup.8).sub.2,
O(CH.sub.2).sub.nN(R.sup.8).sub.2, O(CH.sub.2).sub.nNHR.sup.8,
O(CH.sub.2).sub.n-morpholine, O(CH.sub.2).sub.n-piperazine,
O(CH.sub.2).sub.n-pyrrolidine, O(CH.sub.2).sub.n-piperidine,
O-piperidine, O(CH.sub.2).sub.n-oxopiperazine,
O(CH.sub.2).sub.n-oxomorpholine, O(CH.sub.2).sub.n-oxopyrrolidine,
O(CH.sub.2).sub.nC(CH.sub.3).sub.2(CH.sub.2).sub.nN(R.sup.8).sub.2,
N(CH.sub.2).sub.nC(CH.sub.3).sub.2(CH.sub.2).sub.nN(R.sup.8).sub.2,
O(CH.sub.2).sub.nN(R.sup.8)SO.sub.mA,
O(CH.sub.2).sub.nN(R.sup.8)SO.sub.mAr,
O(CH.sub.2).sub.nN(R.sup.8)SO.sub.mN(R.sup.8).sub.2A,
(CH.sub.2).sub.nN(R.sup.8)SO.sub.mA,
(CH.sub.2).sub.nN(R.sup.8)SO.sub.mAr,
(CH.sub.2).sub.nN(R.sup.8)SO.sub.mN(R.sup.8).sub.2A,
O(CH.sub.2).sub.nSO.sub.mA, O(CH.sub.2).sub.nSO.sub.mAr,
O(CH.sub.2).sub.nSO.sub.mN(R.sup.8)A, (CH.sub.2).sub.nSO.sub.mA,
(CH.sub.2).sub.nSO.sub.mAr, (CH.sub.2).sub.nSO.sub.mN(R.sup.9)A,
and the like. Examples of "heteroaryl" moieties include, but are
not limited to furanyl, thiophenyl, pyranyl, pyrrolyl, imidazolyl,
pyrazolyl, thiazolyl, pyridinyl, pyrazinyl, quinolinyl,
isoquinolinyl, triazolyl, tetrazolyl, oxazolyl, isoxazolyl,
oxadiazolyl, oxo-pyridyl, thiadiazolyl, isothiazolyl, pyridazyl,
pyrimidinyl, benzofuranyl, benzothiophenyl, indolyl, indazolyl, and
the like.
[0142] The nomenclature as used herein for defining compounds,
especially the compounds according to the invention, is in general
based on the rules of the IUPAC-organisation for chemical compounds
and especially organic compounds.
[0143] A further preferred embodiment of the present invention is a
method for the manufacture of a compound according to formula I,
characterized in that [0144] a) an aromatic or heteroaromatic
nitrile according to formula II,
[0144] ##STR00003## [0145] wherein R.sup.1, Wand X are as defined
above, is reacted with a straight or branched chain alcohol
according to formula III,
[0145] HO--R.sup.9 III [0146] wherein R.sup.9 is (CH.sub.2).sub.q
and q is 1-10, [0147] and that the product according to formula
IV,
[0147] ##STR00004## [0148] wherein R.sup.1, R.sup.7, R.sup.9 and X
are as defined above, is then reacted with a compound according to
formula V,
[0148] ##STR00005## [0149] wherein R.sup.2, R.sup.3, R.sup.4,
R.sup.5 and R.sup.6 are as defined above, or [0150] b) a compound
of formula I is isolated and/or treated with an acid or a base, to
obtain the salt thereof.
[0151] A physiologically acceptable salt of a compound according to
formula I can also be obtained by isolating and/or treating the
compound of formula I obtained by the described reaction with an
acid or a base.
[0152] For a further detailed description of the manufacturing
processes, please see also example 1 and the following general
description of the preferred conditions.
[0153] All crude products were subjected to standard chromatography
using solvent mixtures containing methanol, ethanol, isopropanol,
n-hexane, cyclohexane, ethylacetat, dichlormethane or petrol ether,
respectively.
[0154] The compounds of the formula I and also the starting
materials for their preparation are prepared by methods as
described in the examples or by methods known per se, as described
in the literature (for example in standard works, such as
Houben-Weyl, Methoden der Organischen Chemie [Methods of Organic
Chemistry], Georg Thieme Verlag, Stuttgart; Organic Reactions, John
Wiley & Sons, Inc., New York), to be precise under reaction
conditions which are known and suitable for the said reactions. Use
can also be made here of variants which are known per se, but are
not mentioned here in greater detail.
[0155] The starting materials for the claimed process may, if
desired, also be formed in situ by not isolating them from the
reaction mixture, but instead immediately converting them further
into the compounds of the formula I. On the other hand, it is
possible to carry out the reaction stepwise.
[0156] Preferably, the reaction of the compounds is carried out in
the presence of a suitable solvent, that is preferably inert under
the respective reaction conditions. Examples of suitable solvents
are hydrocarbons, such as hexane, petroleum ether, benzene, toluene
or xylene; chlorinated hydrocarbons, such as trichlorethylene,
1,2-dichloroethane, tetrachloromethane, chloroform or
dichloromethane; alcohols, such as methanol, ethanol, isopropanol,
n-propanol, n-butanol or tert-butanol; ethers, such as diethyl
ether, diisopropyl ether, tetrahydrofuran (THE) or dioxane; glycol
ethers, such as ethylene glycol monomethyl or monoethyl ether or
ethylene glycol dimethyl ether (diglyme); ketones, such as acetone
or butanone; amines, such as triethylamine; amides, such as
acetamide, dimethylacetamide, dimethylformamide (DMF) or
N-methylpyrrolidinone (NMP); nitriles, such as acetonitrile;
sulfoxides, such as dimethyl sulfoxide (DMSO); nitro compounds,
such as nitromethane or nitrobenzene; esters, such as ethyl
acetate, or mixtures of the said solvents or mixtures with water.
Polar solvents are in general preferred. Examples for suitable
polar solvents are chlorinated hydrocarbons, alcohols, glycol
ethers, nitriles, amides and sulfoxides or mixtures thereof. More
preferred are 1,4-dioxane, 1-butanol and triethylamine.
[0157] As stated above, the reaction temperature is between about
-100.degree. C. and 300.degree. C., more preferred between 0 and
250.degree. C., preferably including the irradiation in a
microwave, depending on the reaction step and the conditions
used.
[0158] Reaction times are generally in the range between some
minutes and several days, depending on the reactivity of the
respective compounds and the respective reaction conditions.
Suitable reaction times are readily determinable by methods known
in the art, for example reaction monitoring. Based on the reaction
temperatures given above, suitable reaction times generally lie in
the range between 10 min and 48 hrs.
[0159] A base of the formula I can be converted into the associated
acid-addition salt using an acid, for example by reaction of
equivalent amounts of the base and the acid in a preferably inert
solvent, such as ethanol, followed by evaporation. Suitable acids
for this reaction are, in particular, those which give
physiologically acceptable salts. Thus, it is possible to use
inorganic acids, for example sulfuric acid, sulfurous acid,
dithionic acid, nitric acid, hydrohalic acids, such as hydrochloric
acid or hydrobromic acid, phosphoric acids, such as, for example,
orthophosphoric acid, sulfamic acid, furthermore organic acids, in
particular aliphatic, alicyclic, araliphatic, aromatic or
heterocyclic monobasic or polybasic carboxylic, sulfonic or
sulfuric acids, for example formic acid, acetic acid, propionic
acid, hexanoic acid, octanoic acid, decanoic acid, hexadecanoic
acid, octadecanoic acid, pivalic acid, diethylacetic acid, malonic
acid, succinic acid, pimelic acid, fumaric acid, maleic acid,
lactic acid, tartaric acid, malic acid, citric acid, gluconic acid,
ascorbic acid, nicotinic acid, isonicotinic acid, methane- or
ethanesulfonic acid, ethanedisulfonic acid, 2-hydroxyethanesulfonic
acid, benzenesulfonic acid, trimethoxybenzoic acid,
adamantanecarboxylic acid, p-toluenesulfonic acid, glycolic acid,
embonic acid, chlorophenoxyacetic acid, aspartic acid, glutamic
acid, proline, glyoxylic acid, palmitic acid,
parachlorophenoxyisobutyric acid, cyclohexanecarboxylic acid,
glucose 1-phosphate, naphthalenemono- and -disulfonic acids or
laurylsulfuric acid.
[0160] Salts with physiologically unacceptable acids, for example
picrates, can be used to isolate and/or purify the compounds of the
formula I.
[0161] On the other hand, compounds of the formula I can be
converted into the corresponding metal salts, in particular alkali
metal salts or alkaline earth metal salts, or into the
corresponding ammonium salts, using bases (for example sodium
hydroxide, potassium hydroxide, sodium carbonate or potassium
carbonate). Suitable salts are furthermore substituted ammonium
salts, for example the dimethyl-, diethyl- and diisopropylammonium
salts, monoethanol-, diethanol- and diisopropanolammonium salts,
cyclohexyl- and dicyclohexylammonium salts,
dibenzylethylenediammonium salts, furthermore, for example, salts
with arginine or lysine.
[0162] If desired, the free bases of the formula I can be liberated
from their salts by treatment with strong bases, such as sodium
hydroxide, potassium hydroxide, sodium carbonate or potassium
carbonate, so long as no further acidic groups are present in the
molecule. In the cases where the compounds of the formula I have
free acid groups, salt formation can likewise be achieved by
treatment with bases. Suitable bases are alkali metal hydroxides,
alkaline earth metal hydroxides or organic bases in the form of
primary, secondary or tertiary amines.
[0163] Every reaction step described herein can optionally be
followed by one or more working up procedures and/or isolating
procedures. Suitable such procedures are known in the art, for
example from standard works, such as Houben-Weyl, Methadon der
organischen Chemie [Methods of Organic Chemistry],
Georg-Thieme-Verlag, Stuttgart). Examples for such procedures
include, but are not limited to evaporating a solvent, distilling,
crystallization, fractionised crystallization, extraction
procedures, washing procedures, digesting procedures, filtration
procedures, chromatography, chromatography by HPLC and drying
procedures, especially drying procedures in vacuum and/or elevated
temperature.
[0164] Proliferative diseases are caused by a defect in the
intracellular signaling system, or the signal transduction
mechanism of certain proteins. One of the principal mechanisms by
which cellular regulation is effected is through the transduction
of extracellular signals across the membrane, that in turn modulate
biochemical pathways within the cell. Protein phosphorylation
represents one course by which intracellular signals are propagated
from molecule to molecule resulting finally in a cellular response.
These signal transduction cascades are highly regulated and often
overlapping, as evident from the existence of many protein kinases
as well as phosphatases. Phosphorylation of proteins occurs
predominantly at serine, threonine or tyrosine residues, and
protein kinases have therefore been classified by their specificity
of phosphorylation site, i.e. serine/threonine kinases and tyrosine
kinases. Since phosphorylation is such a ubiquitous process within
cells and since cellular phenotypes are largely influenced by the
activity of these pathways, it is currently believed that a number
of disease states and/or diseases are attributable to either
aberrant activation or functional mutations in the molecular
components of kinase cascades. Defects include a change either in
the intrinsic activity or in the cellular concentration of one or
more signaling proteins in the signaling cascade. The cell may
produce a growth factor that binds to its own receptors, resulting
in an autocrine loop, which continually stimulates proliferation.
Mutations or overexpression of intracellular signaling proteins can
lead to spurious mitogenic signals within the cell. Consequently,
considerable attention has been devoted to the characterization of
kinase proteins and compounds that are able to modulate their
activity (for a review see: Weinstein-Oppenheimer et al. Pharma.
&. Therap., 2000, 88: 229-279).
[0165] Tyrosine kinases are a class of enzymes, which catalyse the
transfer of the terminal phosphate of adenosine triphosphate to
tyrosine residues in protein substrates. It is thought that
tyrosine kinases, through substrate phosphorylation, play a crucial
role in signal transduction for a number of cell functions.
Although the precise mechanisms of signal transduction are still
unclear, tyrosine kinases have been shown to be important
contributing factors in cell proliferation, carcinogenesis and cell
differentiation. Tyrosine kinases can be categorised as
receptor-type tyrosine kinases or non-receptor type tyrosine
kinases. Receptor-type tyrosine kinases have an extracellular
portion, a transmembrane portion and an intracellular portion,
while non-receptor type tyrosine kinases are exclusively
intracellular.
[0166] Tyrosine kinases consist of a multiplicity of transmembrane
receptors with different biological activity. Thus, about 20
different subfamilies of receptor-type tyrosine kinases have been
identified. One tyrosine kinase subfamily, known as the HER
subfamily, consists of EGFR, HER2, HERS and HERO. Ligands from this
subfamily of receptors include epithelial growth factor,
TGF-.alpha., amphiregulin, HB-EGF, betacellulin and heregulin.
Another subfamily of these receptor-type tyrosine kinases is the
insulin subfamily, which includes INS-R, IGF-IR and IR-R. The PDGF
subfamily includes the PDGF-.alpha. and -.beta. receptors, CSFIR,
c-kit and FLK-II. In addition, there is the FLK family, which
consists of the kinase insert domain receptor (KDR), foetal liver
kinase-1 (FLK-1), foetal liver kinase-4 (FLK-4) and the fms
tyrosine kinase-1 (flt-1). The PDGF and FLK families are usually
discussed together due to the similarities between the two groups.
For a detailed discussion of receptor-type tyrosine kinases, see
Plowman et al., DN & P 7(6): 334-339, 1994, which is hereby
incorporated by way of reference.
[0167] The non-receptor type tyrosine kinases likewise consist of a
multiplicity of subfamilies, including Src, Frk, Btk, Csk, Abl,
Zap70, Fes/Fps, Fak, Jak, Ack and L1MK. Each of these subfamilies
is further sub-divided into different receptors. For example, the
Src subfamily is one of the largest subfamilies. It includes Src,
Yes, Fyn, Lyn, Lck, Blk, Hck, Fgr and Yrk. The Src subfamily of
enzymes has been linked to oncogenesis. For a more detailed
discussion of non-receptor type tyrosine kinases, see Bolen
Oncogene, 8: 2025-2031 (1993), which is hereby incorporated by way
of reference.
[0168] Both receptor type tyrosine kinases and non-receptor type
tyrosine kinases are involved in cellular signalling pathways
leading to numerous pathogenic conditions, including cancer,
psoriasis and hyperimmune responses. It has been proposed that
various receptor-type tyrosine kinases, and the growth factors
binding to them, play a role in angiogenesis, although some may
promote angiogenesis indirectly (Mustonen and Alitalo, J. Cell
Biol. 129: 895-898, 1995). One of these receptor-type tyrosine
kinases is foetal liver kinase 1, also referred to as FLK-1. The
human analogue of FLK-1 is the kinase insert domain-containing
receptor KDR, which is also known as vascular endothelial cell
growth factor receptor 2 or VEGFR-2, since it binds VEGF with high
affinity. Finally, the murine version of this receptor has also
been called NYK (Oelrichs et al., Oncogene 8(1): 11-15, 1993). VEGF
and KDR are a ligand-receptor pair which plays a vital role in the
proliferation of vascular endothelial cells and the formation and
sprouting of blood vessels, referred to as vasculogenesis and
angiogenesis respectively. Angiogenesis is characterised by
excessive activity of vascular endothelial growth factor (VEGF).
VEGF actually consists of a family of ligands (Klagsburn and
D'Amore, Cytokine & Growth Factor Reviews 7: 259-270, 1996).
VEGF binds the high affinity membrane-spanning tyrosine kinase
receptor KDR and the related fms-like tyrosine kinase-1, also known
as Flt-1 or vascular endothelial cell growth factor receptor 1
(VEGFR-1). Cell culture and gene knockout experiments indicate that
each receptor contributes to different aspects of angiogenesis. KDR
mediates the mitogenic function of VEGF, whereas Flt-1 appears to
modulate non-mitogenic functions, such as those associated with
cellular adhesion. Inhibiting KDR thus modulates the level of
mitogenic VEGF activity. In fact, tumour growth has been shown to
be susceptible to the antiangiogenic effects of VEGF receptor
antagonists (Kim et al., Nature 362, pp. 841-844, 1993).
[0169] Solid tumours can therefore be treated with tyrosine
inhibitors since these tumours depend on angiogenesis for the
formation of the blood vessels that are necessary to support their
growth. These solid tumours include monocytic leukaemia, carcinomas
of the brain, genito-urinary tract, lymphatic system, stomach,
larynx and lung, including lung adenocarcinoma and small cell lung
carcinoma. Further examples include carcinomas in which
overexpression or activation of Raf-activating oncogenes (for
example, K-Ras, Erb-B) is observed. Such carcinomas include
pancreatic and breast carcinoma. Inhibitors of these tyrosine
kinases are therefore suitable for the prevention and treatment of
proliferative diseases caused by these enzymes. The angiogenic
activity of VEGF is not limited to tumours. VEGF accounts for the
angiogenic activity produced in or near the retina in diabetic
retinopathy. This vascular growth in the retina leads to visual
degeneration culminating in blindness. Ocular VEGF mRNA and protein
levels are elevated by conditions such as retinal vein occlusion in
primates and decreased pO.sub.2 levels in mice that lead to
neovascularization. Intraocular injections of anti-VEGF monoclonal
antibodies or VEGF receptor immunofusions inhibit ocular
neovascularization in both primate and rodent models. Irrespective
of the cause of induction of VEGF in human diabetic retinopathy,
inhibition of ocular VEGF is suitable for treating this
disease.
[0170] Expression of VEGF is also significantly increased in
hypoxic regions of animal and human tumours adjacent to areas of
necrosis. In addition, VEGF is upregulated by the expression of the
oncogenes Ras, Raf, Src and mutant p53 (all of which are relevant
in combating cancer). Anti-VEGF monoclonal antibodies inhibit the
growth of human tumours in nude mice. Although the same tumour
cells continue to express VEGF in culture, the antibodies do not
diminish their mitotic rate. Thus, tumour-derived VEGF does not
function as an autocrine mitogenic factor. VEGF therefore
contributes to tumour growth in vivo by promoting angiogenesis
through its paracrine vascular endothelial cell chemotactic and
mitogenic activities. These monoclonal antibodies also inhibit the
growth of typically less well vascularised human colon carcinomas
in athymic mice and decrease the number of tumours arising from
inoculated cells.
[0171] The expression of a VEGF-binding construct of Flk-1, Flt-1,
the mouse KDR receptor homologue truncated to eliminate the
cytoplasmic tyrosine kinase domains but retaining a membrane
anchor, virtually stops the growth of a transplantable glioblastoma
in mice, presumably by the dominant negative mechanism of
heterodimer formation with membrane-spanning endothelial cell VEGF
receptors.
[0172] Embryonic stem cells, which normally grow as solid tumours
in nude mice, do not produce detectable tumours if both VEGF
alleles are knocked out. Taken together, these data indicate the
role of VEGF in the growth of solid tumours. Inhibition of KDR or
Flt-1 is involved in pathological angiogenesis, and these receptors
are suitable for the treatment of diseases in which angiogenesis is
part of the overall pathology, for example inflammation, diabetic
retinal vascularization, as well as various forms of cancer, since
tumour growth is known to be dependent on angiogenesis (Weidner et
al., N. Engl. J. Med., 324, pp. 1-8, 1991).
[0173] Therefore, a preferred embodiment of the present invention
is the use of a compound of the present invention as VEGFR
modulator or its use for the prevention or treatment of
VEGFR-mediated disorders.
[0174] Angiopoietin 1 (Ang1), a ligand for the endothelium-specific
receptor-type tyrosine kinase TIE-2, is a novel angiogenic factor
(Davis et al., Cell, 1996, 87: 1161-1169; Partanen et al., Mol.
Cell. Biol., 12: 1698-1707 (1992); U.S. Pat. Nos. 5,521,073;
5,879,672; 5,877,020; and 6,030,831). The acronym TIE stands for
"tyrosine kinase with Ig and EGF homology domains". TIE is used for
the identification of a class of receptor-type tyrosine kinases,
which are expressed exclusively in vascular endothelial cells and
early haemopoietic cells. TIE receptor kinases are typically
characterised by the presence of an EGF-like domain and an
immunoglobulin (Ig)-like domain, which consists of extracellular
fold units stabilised by disulfide bridge bonds between the chains
(Partanen et al. Curr. Topics Microbiol. Immunol., 1999, 237:
159-172). In contrast to VEGF, which exerts its function during the
early stages of vascular development, Ang1 and its receptor TIE-2
act during the later stages of vascular development, i.e. during
vascular transformation (transformation relates to the formation of
a vascular lumen) and maturing (Yancopoulos et al., Cell, 1998,
93:661-664; Peters, KG., Circ. Res., 1998, 83(3): 342-3; Suri et
al., Cell 87, 1171-1180 (1996)).
[0175] Accordingly, it would be expected that inhibition of TIE-2
should interrupt the transformation and maturing of a new vascular
system initiated by angiogenesis and should thus interrupt the
angiogenesis process. Furthermore, inhibition at the kinase
domain-binding site of VEGFR-2 would block phosphorylation of
tyrosine residues and serve to interrupt initiation of
angiogenesis. It must therefore be assumed that inhibition of TIE-2
and/or VEGFR-2 should prevent tumour angiogenesis and serve to slow
or completely eliminate tumour growth. Accordingly, treatment of
cancer and other diseases associated with inappropriate
angiogenesis could be provided.
[0176] The present invention also therefore relates to methods for
the regulation, modulation or inhibition of TIE-2 for the
prevention and/or treatment of diseases associated with unregulated
or disturbed TIE-2 activity. In particular, the compounds according
to the invention can also be employed in the treatment of certain
forms of cancer. Furthermore, the compounds according to the
invention can be used to provide additive or synergistic effects in
certain existing cancer chemotherapies and/or can be used to
restore the efficacy of certain existing cancer chemotherapies and
radio-therapies.
[0177] The protein kinase PKB (also known as AKT and RAC-PK) is a
member of the AKT/PKB family of serine/threonine kinases and has
been shown to be involved in a diverse set of signalling pathways
in human malignancy (Nicholson et al., Cell. Signal., 2002, 14,
381-395). PKB, like other members of the AKT/PKB family, is located
in the cytosol of unstimulated cells and translocates to the cell
membrane following stimulation. PKB translocation can be activated
by a number of ligands, including platelet derived growth factor,
epidermal growth factor, basic fibroblast growth factor, cellular
stress, such as, for example, heat shock and hyperosmolarity, as
well as insulin (Bos, Trends Biochem. Sci., 1995, 20, 441-442), and
other studies have shown that this activation is through PI3 kinase
which is wortmannin sensitive (Franke et al., Science, 1997, 275,
665-668). Once localised to the plasma membrane, PKB has been shown
to mediate several functions within the cell, including apoptosis,
the metabolic effects of insulin, induction of differentiation
and/or proliferation, protein synthesis and stress responses
(Alessi and Cohen, Curr. Opin. Genet. Dev., 1998, 8: 55-62;
Downward, Curr. Opin. Cell Biol., 1998, 10, 262-267).
[0178] PKB was cloned independently in 1991 by three groups
(Bellacosa et al., Science, 1991, 254, 274-277; Coffer and
Woodgett, Eur. J. Biochem., 1991, 201, 475-481; Jones et al., Cell
Regul., 1991, 2, 1001-1009), but its association with primary human
gastric carcinoma was recognised as early as 1987 (Staal et al.,
Proc. Natl. Acad. Sci. USA, 1987, 84, 5034-5037). Sequencing of
PKB.alpha. revealed homology within the kinase domains to the PKA
(about 68%) and PKC isozymes (about 73%) (Jones et al., Proc. Natl.
Acad. Sci. U.S.A., 1991, 88, 4171-5), a fact that lead to its
renaming as PKB. There are three cellular isoforms of PKB and two
splice variants (PKB.alpha., .beta., .gamma., .beta..sub.1,
.gamma..sub.1; Brazil et al. Trends in Bio Sci, 2001, 26, 657-663).
PKB.alpha. was found to be amplified or overexpressed in gastric
adenocarcinomas and in a breast cancer cell line (Staal et al.,
Proc. Natl. Acad. Sci. U.S.A., 1987, 84, 5034-7; Jones et al., Cell
Regul., 1991, 2, 1001-9). PKB.beta. is amplified or overexpressed
in 3% of breast (Bellacosa et al., Int. J. Cancer, 1995 64, 280-5),
12% of pancreatic (Cheng et al., Proc. Natl. Acad. Sol. U.S.A.,
1996, 93, 3636-41) and 15% of ovarian cancers (Bellacosa et al.,
Int. J. Cancer, 1995, 64, 280-5; Cheng et al., Proc. Natl. Acad.
Sci. U.S.A., 1992, 89, 9267-71).
[0179] PKB.gamma. is overexpressed in oestrogen receptor-deficient
breast cancer and in androgen-independent prostate cell lines
(Nakatani et al., J. Biol. Chem. 1999, 274, 21528-32). It has been
proposed that PKB is a gene, which is involved in chromosomal
rearrangement at chromosome band 1402. This locus is known to
undergo rearrangement in human T-cell malignancies, such as, for
example, prolymphocytic leukaemias and mixed lineage childhood
leukaemias (Staal et al., Genomics, 1988, 2, 96-98).
[0180] PKB also plays a role in the prevention of "programmed cell
death" or apoptosis by inhibitory phosphorylation of ASK-1, Bad,
Caspase9 and FKHR (for review see Nicholson et al., Cell Signalling
2001, 14, 281-395). It has been shown that PKB provides a survival
signal (for review see Lawlor et al., J. of Cell Science 2001, 114,
2903-2910) to cells in order to protect them from a number of
agents, including UV radiation (Dudek et al., Science, 1997, 275,
661-665), withdrawal of IGF1 from neuronal cells, detachment from
the extracellular matrix, stress and heat shock (Alessi and Cohen,
Curr. Opin. Genet. Dev., 1998, 8: 55-62).
[0181] The dual-specific phosphatase PTEN (phosphatase and tensin
homologue deleted on chromosome ten) increases the Ptdlns(3, 4,
5)P.sub.3 level in the cell by dephosphorylation of Ptdlns(3, 4,
5)P.sub.3. Ptdlns(3, 4, 5)P.sub.3 binds to the PH domain
(Pleckstrin homology domain) of PKB. This binding is an essential
step for membrane translocation and activation of PKB. PTEN is a
tumour suppressor gene mutated in a large proportion of
glioblastoma and melanoma cell lines, advanced prostate carcinomas
and endometrial carcinomas. Furthermore, it is deleted in >80%
of patients with hereditary conditions, such as, for example,
Cowden's disease, Lhermitte-Duclose disease and Bannayan-Zonana
Syndrome. The patients display a number of similar features,
including multiple benign tumours (harmatomas) and increased
susceptibility to breast and thyroid malignancies (Di Cristofano et
al. Cell, 2000, 100, 387-390).
[0182] Cell lines derived from PTEN.sup.+/- heterozygous mice
(PTEN.sup.-/- heterozygous mice are not viable) show increased
Ptdlns(3, 4, 5)P.sub.3 levels paralleled by increased PKB activity,
with concomitant decreased sensitivity to apoptosis (Di Christofano
et al. Nat. Genet. 1998, 19, 348-355; Stambolic et al., Cell, 1998,
95, 29-39, Myers et al., Proc. Natl. Acad. Si. U.S.A., 1998, 96:
13513-13518). PKB is also able to promote cell cycle progression by
inhibiting p21 cell cycle inhibitor (Zhou et al.; Nat. Cell Biol.,
2002, 3: 245-252). These findings may explain the overexpression of
PKB observed in cancer cells, which allows preferential survival
and proliferation of the carcinomas by avoiding the normal
progression to apoptosis.
[0183] Therefore, a preferred embodiment of the present invention
is the use of a compound of the present invention as PKB modulator
or its use for the prevention or treatment of PKB-mediated
disorders.
[0184] Some of the most common mutations leading to upregulation of
mitogenic signals and proliferative diseases occur in genes
encoding the protein known as Ras, a G-protein that is activated
when bound to GTP, and inactivated when bound to GDP. The
above-mentioned growth factor receptors, and many other mitogenic
receptors, when activated, lead to Ras being converted from the
GDP-bound state to the GTP-bound state. This signal is an absolute
prerequisite for proliferation in most cell types. Defects in this
signaling system, especially in the deactivation of the Ras-GTP
complex, are common in cancers, and lead to the signaling cascade
below Ras being chronically activated.
[0185] Activated Ras leads in turn to the activation of a cascade
of serine/threonine kinases. One of the groups of kinases known to
require an active Ras-GTP for its own activation is the Raf
family.
[0186] The present invention therefore relates to the compounds of
the present invention as inhibitors of Raf kinases. Protein
phosphorylation is a fundamental process for the regulation of
cellular functions. The coordinated action of both protein kinases
and phosphatases controls the degrees of phosphorylation and,
hence, the activity of specific target proteins. One of the
predominant roles of protein phosphorylation is in signal
transduction, where extracellular signals are amplified and
propagated by a cascade of protein phosphorylation and
dephosphorylation events, for example in the p21.sup.ras/Raf
pathway.
[0187] The p21.sup.ras gene was discovered as an oncogene of the
Harvey (H-Ras) and Kirsten (K-Ras) rat sarcoma viruses. In humans,
characteristic mutations in the cellular Ras gene (c-Ras) have been
associated with many different types of cancers. These mutant
alleles, which render Ras constitutively active, have been shown to
transform cells, such, for example, as the murine cell line NIH
3T3, in culture.
[0188] The p21.sup.ras oncogene is a major contributor to the
development and pro-gression of human solid carcinomas and is
mutated in 30% of all human carcinomas (Bolton et al. (1994) Ann.
Rep. Med. Chem., 29, 165-74; Bos. (1989) Cancer Res., 49: 4682-9).
In its normal, unmutated form, the Ras protein is a key element of
the signal transduction cascade directed by growth factor receptors
in almost all tissues (Avruch et al. (1994) Trends Biochem. Sci.,
19: 279-83).
[0189] Biochemically, Ras is a guanine nucleotide binding protein
and cycling between a GTP-bound activated and a GDP-bound resting
form is strictly controlled by Ras endogenous GTPase activity and
other regulatory proteins. The Ras gene product binds to guanine
triphosphate (GTP) and guanine diphosphate (GDP) and hydrolyses GTP
to GDP. Ras is active in the GTP-bound state. In the Ras mutants in
cancer cells, the endogenous GTPase activity is reduced and the
protein consequently transmits constitutive growth signals to
downstream effectors, such as, for example, the enzyme Raf kinase.
This leads to the cancerous growth of the cells, which carry these
mutants (Magnuson et al. (1994) Semin. Cancer Biol., 5, 247-53).
The Ras proto-oncogene requires a functionally intact c-Raf-1
proto-oncogene in order to transduce growth and differentiation
signals initiated by receptor and non-receptor-type tyrosine
kinases in higher eukaryotes.
[0190] Activated Ras is necessary for the activation of the c-Raf-1
proto-oncogene, but the biochemical steps through which Ras
activates the Raf-1 protein (Ser/Thr) kinase are now well
characterised. It has been shown that inhibiting the effect of
active Ras by inhibiting the Raf kinase signalling pathway by
administration of deactivating antibodies to Raf kinase or by
co-expression of dominant negative Raf kinase or dominant negative
MEK (MAPKK), the substrate of Raf kinase, leads to reversion of
transformed cells to the normal growth phenotype (see: Daum et al.
(1994) Trends Biochem. Sci., 19, 474-80; Fridman et al. (1994) J.
Biol. Chem., 269, 30105-8. Kolch et al. (1991) Nature, 349, 426-28
and for a review Weinstein-Oppenheimer et al. Pharm. & Therap.
(2000), 88, 229-279).
[0191] Similarly, inhibition of Raf kinase (by antisense
oligodeoxynucleotides) has been correlated in vitro and in vivo
with inhibition of the growth of a variety of human tumour types
(Monia et al., Nat. Med. 1996, 2, 668-75).
[0192] Raf serine- and threonine-specific protein kinases are
cytosolic enzymes that stimulate cell growth in a variety of cell
systems (Rapp, U. R., et al. (1988) in The Oncogene Handbook; T.
Curran, E. P. Reddy and A. Skalka (eds.) Elsevier Science
Publishers; The Netherlands, pp. 213-253; Rapp, U. R., et al.
(1988) Cold Spring Harbor Sym. Quant. Biol. 53: 173-184; Rapp, U.
R., et al. (1990) Inv Curr. Top. Microbial. Immunol. Potter and
Melchers (eds.), Berlin, Springer-Verlag 166: 129-139).
[0193] Three isozymes have been characterised: C-Raf (Raf-1)
(Bonner, T. I., et al. (1986) Nucleic Acids Res. 14: 1009-1015).
A-Raf (Beck, T. W., et al. (1987) Nucleic Acids Res. 15: 595-609)
and B-Raf (Qkawa, S., et al. (1998) Mol. Cell. Biol. 8: 2651-2654;
Sithanandam, G. et al. (1990) Oncogene: 1775). These enzymes differ
in their expression in various tissues. Raf-1 is expressed in all
organs and in all cell lines that have been examined, and A- and
B-Raf are expressed in urogenital and brain tissues respectively
(Storm, S. M. (1990) Oncogene 5: 345-351).
[0194] Raf genes are proto-oncogenes: they can initiate malignant
transformation of cells when expressed in specifically altered
forms. Genetic changes that lead to oncogenic activation generate a
constitutively active protein kinase by removal of or interference
with an N-terminal negative regulatory domain of the protein
(Heidecker, G., et al. (1990) Mol. Cell. Biol. 10: 2503-2512; Rapp,
U. R., et al. (1987) in Oncogenes and Cancer; S. A. Aaronson, J.
Bishop, T. Sugimura, M. Terada, K. Toyoshima and P. K. Vogt (eds.)
Japan Scientific Press, Tokyo). Microinjection into NIH 3T3 cells
of oncogenically activated, but not wild-type, versions of the Raf
protein prepared with Escherichia coli expression vectors results
in morphological transformation and stimulates DNA synthesis (Rapp,
U. R., et al. (1987) in Oncogenes and Cancer; S. A. Aaronson, J.
Bishop, T. Sugimura, M. Terada, K. Toyoshima and P. K. Vogt (ed.)
Japan Scientific Press, Tokyo; Smith, M. R., et al. (1990) Mol.
Cell. Biol. 10: 3828-3833).
[0195] Consequently, activated Raf-1 is an intracellular activator
of cell growth. Raf-1 protein serine kinase is a candidate for the
downstream effector of mitogen signal transduction, since Raf
oncogenes overcome growth arrest resulting from a block of cellular
Ras activity due either to a cellular mutation (Ras revertant
cells) or microinjection of anti-Ras antibodies (Rapp, U. R., et
al. (1988) in The Oncogene Handbook, T. Curran, E. P. Reddy and A.
Skalka (ed.), Elsevier Science Publishers; The Netherlands, pp.
213-253; Smith, M. R., et al. (1986) Nature (London) 320:
540-543).
[0196] C-Raf function is required for transformation by a variety
of membrane-bound oncogenes and for growth stimulation by mitogens
contained in serums (Smith, M. R., et al. (1986) Nature (London)
320: 540-543). Raf-1 protein serine kinase activity is regulated by
mitogens via phosphorylation (Morrison, D. K., et al. (1989) Cell
58: 648-657), which also effects sub-cellular distribution (Olah,
Z., et al. (1991) Exp. Brain Res. 84: 403; Rapp, U. R., et al.
(1988) Cold Spring Harbor Sym. Quant. Biol. 53: 173-184. Raf-1
activating growth factors include platelet-derived growth factor
(PDGF) (Morrison, D. K., et al. (1988) Proc. Natl. Acad. Sci. USA
85: 8855-8859), colony-stimulating factor (Baccarini, M., et al.
(1990) EMBO J. 9: 3649-3657), insulin (Blackshear, P. J., et al.
(1990) J. Biol. Chem. 265: 12115-12118), epidermal growth factor
(EGF) (Morrison, R. K., et al. (1988) Proc. Natl. Acad. Sci. USA
85: 8855-8859), interleukin-2 (Turner, B. C., et al. (1991) Proc.
Natl. Acad. Sci. USA 88:1227) and interleukin-3 and granulocyte
macrophage colony-stimulating factor (Carroll, M. P., et al. (1990)
J. Biol. Chem. 265: 19812-19817).
[0197] After mitogen treatment of cells, the transiently activated
Raf-1 protein serine kinase translocates to the perinuclear area
and the nucleus (Olah, Z., et al. (1991) Exp. Brain Res. 84: 403;
Rapp, U. R., et al. (1988) Cold Spring Harbor Sym. Quant. Biol. 53:
173-184). Cells containing activated Raf are altered in their
pattern of gene expression (Heidecker, G., et al. (1989) in Genes
and signal transduction in multistage carcinogenesis, N. Colburn
(ed.), Marcel Dekker, Inc., New York, pp. 339-374) and Raf
oncogenes activate transcription from Ap-I/PEA3-dependent promoters
in transient transfection assays (Jamal, S., et al. (1990) Science
344: 463-466; Kaibuchi, K., et al. (1989) J. Biol. Chem. 264:
20855-20858; Wasylyk, C., et al. (1989) Mol. Cell. Biol. 9:
2247-2250).
[0198] There are at least two independent pathways for Raf-1
activation by extracellular mitogens: one involving protein kinase
C (KC) and a second initiated by protein tyrosine kinases
(Blackshear, P. J., et al. (1990) J. Biol. Chem. 265:12131-12134;
Kovacina, K. S., et al. (1990) J. Biol. Chem. 265: 12115-12118;
Morrison, D. K., et al. (1988) Proc. Natl. Acad. Sci. USA 85:
8855-8859; Siegel, J. N., et al. (1990) J. Biol. Chem. 265:
18472-18480; Turner, B. C., et al. (1991) Proc. Natl. Acad. Sci.
USA 88: 1227). In each case, activation involves Raf-1 protein
phosphorylation. Raf-1 phosphorylation may be a consequence of a
kinase cascade amplified by autophosphorylation or may be caused
entirely by autophosphorylation initiated by binding of a putative
activating ligand to the Raf-1 regulatory domain, analogous to PKC
activation by diacylglycerol (Nishizuka, Y. (1986) Science 233:
305-312).
[0199] MAPK/ERK Kinase ("MEK") enzymes are dual specificity kinases
involved in, for example, immunomodulation, inflammation, and
proliferative diseases such as cancer and restenosis.
[0200] These kinases of the Raf family activate MEK (e.g., MEK1 and
MEK2) which then activates the MAP kinase, ERK (ERK1 and ERK2).
Activation of MAP kinase by mitogens appears to be essential for
proliferation, constitutive activation of this kinase is sufficient
to induce cellular transformation. Blockade of downstream Ras
signaling, for example by use of a dominant negative Raf-1 protein,
can completely inhibit mitogenesis, whether induced from cell
surface receptors or from oncogenic Ras mutants. Although Ras is
not itself a protein kinase, it participates in the activation of
Raf and other kinases, most likely through a phosphorylation
mechanism. Once activated, Raf and other kinases phosphorylate MEK
on two closely adjacent serine residues, S.sup.218 and S.sup.222 in
the case of MEK1, which are the prerequisite for activation of MEK
as a kinase. MEK in turn phosphorylates MAP kinase on both a
tyrosine, Y.sup.185, and a threonine residue, T.sup.183, separated
by a single amino acid. This double phosphorylation activates MAP
kinase at least 100-fold. Activated MAP kinase can then catalyze
the phosphorylation of a large number of proteins, including
several transcription factors and other kinases. Many of these MAP
kinase phosphorylations are mitogenically activating for the target
protein, such as a kinase, a transcription factor, or another
cellular protein. In addition to Raf-1 and MEKK, other kinases
activate MEK, and MEK itself appears to be a signal integrating
kinase. Current understanding is that MEK is highly specific for
the phosphorylation of MAP kinase. In fact, no substrate for MEK
other than the MAP kinase, ERK, has been demonstrated to date and
MEK does not phosphorylate peptides based on the MAP kinase
phosphorylation sequence, or even phosphorylate denatured MAP
kinase. MEK also appears to associate strongly with MAP kinase
prior to phosphorylating it, suggesting that phosphorylation of MAP
kinase by MEK may require a prior strong interaction between the
two proteins. Both this requirement and the unusual specificity of
MEK are suggestive that it may have enough difference in its
mechanism of action to other protein kinases that selective
inhibitors of MEK, possibly operating through allosteric mechanisms
rather than through the usual blockade of the ATP binding site, may
be found.
[0201] It has been found that the compounds of the present
invention are inhibitors of MEK and are useful in the treatment of
a variety of proliferative disease states, such as conditions
related to the hyperactivity of MEK, as well as diseases modulated
by the MEK cascade.
[0202] Selective MEK1 or MEK2 inhibitors are those compounds, which
inhibit the MEK1 or MEK2 enzymes, respectively, without
substantially inhibiting other enzymes such as MKK3, PKC, Cdk2A,
phosphorylase kinase, EGF and C-src. In general, a selective MEK1
or MEK2 inhibitor has an IC.sub.50 for MEK1 or MEK2 that is at
least one-fiftieth ( 1/50) that of its IC.sub.50 for one of the
above-named other enzymes. Preferably, a selective inhibitor has an
IC.sub.50 that is at least 1/100, more preferably 1/500, and even
more preferably 1/1000, 1/5000, or less than that of its IC.sub.50
or one or more of the above-named enzymes.
[0203] The identification of small compounds, which specifically
inhibit, regulate and/or modulate signal transduction of protein
kinases and can be used as medicaments for the treatment of various
diseases is therefore desirable and an aim of the present
invention.
[0204] The disclosed compounds are useful as both prophylactic and
therapeutic treatments for disorders or conditions related to the
change of activity of one or more of the above-mentioned kinases,
especially the hyperactivity of MEK, as well as diseases or
conditions modulated by said kinase cascades.
[0205] Thus, a further preferred embodiment of the present
invention is the use of a compound of the formula I for the
preparation of a medicament for the treatment and/or prevention of
disorders.
[0206] The invention therefore also relates to compounds of the
formula I and the physiologically acceptable salts, derivatives,
prodrugs, solvates and stereoisomers thereof, including mixtures
thereof in all ratios as medicaments.
[0207] A further preferred embodiment of the present invention is
the use of the compounds according to the invention for the
manufacture of a medicament for the treatment and/or prevention of
disorders, which are caused, mediated and/or propagated by protein
kinases.
[0208] A further preferred embodiment of the present invention is
the use of the compounds according to the invention for the
manufacture of a medicament for the treatment and/or prevention of
disorders, which are caused, mediated and/or propagated by protein
kinases, characterized in that the protein kinases are MEK1 or
MEK2.
[0209] Usually, the disorders discussed herein are divided into two
groups, hyperproliferative and non-hyperproliferative disorders. In
this context, infection or infectious diseases, psoriasis,
arthritis, inflammation, endometriosis, scarring, begnin prostatic
hyperplasia, immunological diseases, autoimmune diseases and
immunodeficiency diseases are to be regarded as non-cancerous
disorders, of which infection, arthritis, inflammation,
immunological diseases, autoimmune diseases and immunodeficiency
diseases are usually regarded as non-hyperproliferative disorders.
In this context, brain cancer, lung cancer, squamous cell cancer,
bladder cancer, gastric cancer, pancreatic cancer, hepatic cancer,
renal cancer, colorectal cancer, breast cancer, head cancer, neck
cancer, oesophageal cancer, gynaecological cancer, thyroid cancer,
lymphoma, chronic leukaemia and acute leukaemia are to be regarded
as cancerous disorders, all of which are usually regarded as
hyperproliferative disorders.
[0210] Thus, a preferred embodiment of the present invention is the
use of a compound of the formula I for the preparation of a
medicament for the treatment and/or prevention of disorders,
characterized in that the disorders are selected from the group
consisting of hyperproliferative and non-hyperproliferative
disorders.
[0211] In a preferred embodiment of the present invention the
disorder is non-cancerous.
[0212] Therefore, the compounds of the formula I can be used for
the preparation of a medicament for the treatment and/or prevention
of disorders, which are selected from the group consisting of
psoriasis, arthritis, rheumatoid arthritis, inflammation,
endometriosis, scarring, Helicobacter pylori Influenza A infection,
HIV infection, hepatitis (B) virus (HBV) infection, human papilloma
virus (HPV) infection, cytomegalovirus (CMV) infection, and
Epstein-Barr virus (EBV) infection, begnin prostatic hyperplasia,
immunodeficiency diseases, autoimmune disease, immunological
diseases, chronic obstructive pulmonary disease, asthma,
inflammatory bowel disease, fibrosis, atherosclerosis, restenosis,
vascular disease, cardiovascular disease, stroke (such as acute
focal ischemic stroke and global cerebral ischemia), heart failure,
cystic fibrosis, hepatomegaly, cardiomegaly, septic shock,
Alzheimer's disease, chronic or neuropathic pain, renal disease and
angiogenesis disorders, mesangial cell proliferative disorders,
diabetic nephropathy, diabetic retinopathy, malignant
nephrosclerosis, thrombotic microangiopathy syndromes, xenograft
(cell(s), skin, limb, organ or bone marrow transplant) rejection,
glomerulopathies, metabolic disorders and neurodegenerative
diseases.
[0213] Infections according the invention include, but are not
limited to infections caused by pathogenic microorganisms, such as
bacteria, fungi, viruses and protozoans, for example influenza
(Pleschka, S. et al. Nature Cell Biol. 2001, 3, page 301-305),
retroviruses, for example HIV infection (Yang, X. et al. J. Biol.
Chem. 1999, 274, page 27981-27988; Popik, W et al Mol Cell Biol.
1996, 16, page 6532-6541), Hepatitis B (Benn, J et al., Proc. Natl.
Acad. Sci. 1995, 92, page 11215-11219), Hepatitis C (Aoki et al. J.
Virol. 2000, 74, page 1736-1741), papillomavirus, parainfluenza,
rhinoviruses, adenoviruses, Heliobacter pylori, and viral and
bacterial infections of the skin (e.g. cold sores, warts,
chickenpox, molluscum, contagiosum, herpes zoster, boils,
cellulitis, erysipelas, impetigo, tinea, Althlete's foot and
ringworm).
[0214] Furthermore, a preferred embodiment of the present invention
is the use of a compound of the formula I for the manufacture of a
medicament for the treatment and/or prevention of disorders,
characterized in that the disorders are selected from the group
consisting of hyperproliferative disorders.
[0215] There are many disorders associated with a dysregulation of
cellular proliferation. The conditions of interest include, but are
not limited to, the following conditions. The subject compounds are
useful in the treatment of a variety of conditions where there is
proliferation and/or migration of smooth muscle cells, and/or
inflammatory cells into the intimal layer of a vessel, resulting in
restricted blood flow through that vessel, e.g., neointimal
occlusive lesions. Occlusive vascular conditions of interest
include atherosclerosis, graft coronary vascular disease after
transplantation, vein graft stenosis, peri-anastomatic prothetic
graft stenosis, restenosis after angioplasty or stent placement,
and the like.
[0216] Furthermore, the compounds of the formula I preferably show
anti-angiogenic properties. Thus, compounds of the present
invention can be advantageously employed in the treatment of one or
more diseases afflicting mammals which are characterized by
cellular proliferation in the area of disorders associated with
neo-vascularization and/or vascular permeability including blood
vessel proliferative disorders including arthritis and restenosis;
fibrotic disorders including hepatic cirrhosis and atherosclerosis;
mesangial cell proliferative disorders include glomerulonephritis,
diabetic nephropathy, malignant nephrosclerosis, thrombotic
microangiopathy syndromes, organ transplant rejection and
glomerulopathies; and metabolic disorders include psoriasis,
diabetes mellitus, chronic wound healing, inflammation and
neurodegenerative diseases.
[0217] The process of angiogenesis is the development of new blood
vessels, generally capillaries, from pre-existing vasculature.
Angiogenesis is defined as involving (i) activation of endothelial
cells; (ii) increased vascular permeability; (iii) subsequent
dissolution of the basement membrane and extravisation of plasma
components leading to formation of a provisional fibrin gel
extracellular matrix; (iv) proliferation and mobilization of
endothelial cells; (v) reorganization of mobilized endothelial
cells to form functional capillaries; (vi) capillary loop
formation; and (vii) deposition of basement membrane and
recruitment of perivascular cells to newly formed vessels. Normal
angiogenesis is activated during tissue growth, from embryonic
development through maturity, and then enters a period of relative
quiescence during adulthood.
[0218] Normal angiogenesis is also activated during wound healing,
and at certain stages of the female reproductive cycle.
Inappropriate or pathological angiogenesis has been associated with
several disease states including various retinopathies; ischemic
disease; atherosclerosis; chronic inflammatory disorders;
rheumatoid arthritis, and cancer. The role of angiogenesis in
disease states is discussed, for instance, in Fan et al., Trends in
Pharmacol Sci. 16:54 66; Shawver et al., DOT Vol. 2, No. 2 Feb.
1997; Folkmann, 1995, Nature Medicine 1:27-31.
[0219] In cancer the growth of solid tumors has been shown to be
angiogenesis dependent. (see Folkmann, J., J. Nat'l. Cancer Inst.,
1990, 82, 4-6.) Consequently, the targeting of pro-angiogenic
pathways is a strategy being widely pursued in order to provide new
therapeutics in these areas of great, unmet medical need.
[0220] Several protein kinases are involved in angiogenic
processes. Endothelial growth factors (e.g. vascular endothelial
growth factor VEGF) activate receptor tyrosine kinases (e.g.
VEGFR-2) and signal through the Ras/Raf/Mek/Erk kinase cascade.
Activation of VEGFR-2 by VEGF is a critical step in the signal
transduction pathway that initiates tumor angiogenesis. VEGF
expression may be constitutive to tumor cells and can also be
upregulated in response to certain stimuli. One such stimuli is
hypoxia, where VEGF expression is upregulated in both tumor and
associated host tissues. The VEGF ligand activates VEGFR-2 by
binding with its extracellular VEGF binding site. This leads to
receptor dimerization of VEGFRs and autophosphorylation of tyrosine
residues at the intracellular kinase domain of VEGFR-2. The kinase
domain operates to transfer a phosphate from ATP to the tyrosine
residues, thus providing binding sites for signaling proteins
downstream of VEGFR-2 leading ultimately to initiation of
angiogenesis (McMahon, G., The Oncologist, Vol. 5, No. 90001, 3-10,
April 2000).
[0221] Mice with a targeted disruption in the B-raf gene die of
vascular defects during development (Wojnowski, L. et al. 1997,
Nature genetics 16, page 293-296). These mice show defects in the
formation of the vascular system and in angiogenesis e.g. enlarged
blood vessels and increased apoptotic death of differentiated
endothelial cells.
[0222] Diseases where there is hyperproliferation and tissue
remodelling or repair or reproductive tissue, e.g., uterine,
testicular and ovarian carcinomas, endometriosis, squamous and
glandular epithelial carcinomas of the cervix, etc. are reduced in
cell number by administration of the subject compounds. The growth
and proliferation of neural cells is also of interest.
[0223] Tumor cells are characterized by uncontrolled growth,
invasion to surrounding tissues, and metastatic spread to distant
sites. Growth and expansion requires an ability not only to
proliferate, but also to down-modulate cell death (apoptosis) and
activate angiogenesis to produce a tumor neovasculature.
[0224] A preferred embodiment of the present invention is the use
of a compound of the formula I for the manufacture of a medicament
for the treatment and/or prevention of disorders, characterized in
that the disorder is cancer, whether solid or hematopoietic.
[0225] Therefore, a preferred embodiment of the present invention
is the use of a compound of the general formula I for the
manufacture of a medicament for the treatment and/or prevention of
disorders, characterised in that the disorders are selected from
the group consisting of carcinomas, e.g. melanoma, brain cancer,
lung cancer, non-small cell lung carcinoma, transitional and
squamous cell urinary carcinoma, bladder cancer, gastric cancer,
pancreatic cancer, colon cancer, duodenal cancer, ductal cancer,
endometrial cancer, stomach cancer, colorectal cancer, hepatic
cancer, renal cancer, breast cancer, head cancer, neck cancer,
oesophageal cancer, gynaecological cancer, ovarian cancer, uterine
cancer, prostate cancer, thyroid cancer, dysplastic oral mucosa,
polyposis, invasive oral cancer, etc.; neurological malignancies;
e.g. neuroplastoma, gliomas, etc; hematological malignancies, e.g.,
childhood acute leukaemia, non-Hodgkin's lymphomas, chronic
lymphocytic leukaemia, malignant cutaneous T-cells, mycosis
fungoides, non-MF cutaneous T-cell-lymphoma, lymphomatoid
papulosis, T-cell rich cutaneous lymphoid hyperplasia, bullous
pemphigoid, discoid lupus erythematosus, lichen planus, etc.; and
the like.
[0226] Tumors of neural tissue are of particular interest, e.g.,
gliomas, neuromas, etc. Some cancers of particular interest include
breast cancers, which are primarily adenocarcinoma subtypes. Ductal
carcinoma in situ is the most common type of noninvasive breast
cancer. In DCIS, the malignant cells have not metastasized through
the walls of the ducts into the fatty tissue of the breast.
Infiltration (or invasive) ductal carcinoma (IDC) has metastasized
through the wall of the duct and invaded the fatty tissue of the
breast. Infiltrating (or invasive) lobular carcinoma (ILC) is
similar to IDC, in that it has the potential to metastasize
elsewhere in the body. About 10% to 15% of invasive breast cancers
are invasive lobular carcinomas.
[0227] Also of interest is non-small cell lung carcinoma. Non-small
cell lung cancer (NSCLC) is made up of three general subtypes of
lung cancer. Epidermoid carcinoma (also called squamos cell
carcinoma) usually starts in one of the larger bronchial tubes and
grows relatively slowly. The size of these tumors can range from
very small to quite large. Adenocarcinoma starts growing near the
outside surface of the lung and may vary in both size and growth
rate. Some slowly growing adenocarcinomas are described as alveolar
cell cancer. Large cell carcinoma starts near the surface of the
lung, grows rapidly, and the growth is usually fairly large when
diagnosed. Other less common forms of lung cancer are carcinoid,
cylindroma, mucoepidermoid, and malignant mesothelioma.
[0228] Melanoma is a malignant tumor of melanocytes. Although most
melanomas arise in the skin, they also may arise from mucosal
surfaces or at other sites to which neural crest cells migrate.
Melanoma occurs predominantly in adults, and more than half of the
cases arise in apparently normal areas of the skin. Prognosis is
affected by clinical and histological factors and by anatomic
location of the lesion. Thickness and/or level of invasion of the
melanoma, mitotic index, tumor infiltrating lymphocytes, and
ulceration or bleeding at the primary site affect the prognosis.
Clinical staging is based on whether the tumor has spread to
regional lymph nodes or distant sites. For disease, clinically
confined to the primary site, the greater the thickness and depth
of local invasion of the melanoma, the higher the chance of lymph
node metastases and the worse the prognosis. Melanoma can spread by
local extension (through lymphatics) and/or by hematogenous routes
to distant sites. Any organ may be involved by metastases, but
lungs and liver are common sites.
[0229] Other hyperproliferative diseases of interest relate to
epidermal hyperproliferation, tissue, remodeling and repair. For
example, the chronic skin inflammation of psoriasis is associated
with hyperplastic epidermal keratinocycles as well as infiltrating
mononuclear cells, including CD4+ memory T cells, neutrophils and
macrophages.
[0230] The proliferation of immune cells is associated with a
number of autoimmune and lymphoproliferative disorders. Diseases of
interest include multiple sclerosis, rheumatoid arthritis and
insulin dependent diabetes mellitus. Evidence suggests that
abnormalities in apoptosis play a part in the pathogenesis of
systemic lupus erythematosus (SLE). Other lymphoproliferative
conditions the inherited disorder of lymphocyte apoptosis, which is
an autoimmune lymphoproliferative syndrome, as well as a number of
leukemia's and lymphomas. Symptoms of allergies to environmental
and food agents, as well as inflammatory bowel disease, may also be
alleviated by the compounds of the invention.
[0231] For use in the subject methods, the subject compounds may be
formulated with pharmaceutically active agents other than the
compounds according to the invention, particularly other
anti-metastatic, antitumor or anti-angiogenic agents. Angiostatic
compounds of interest include angiostatin, enclostatin, carboxy
terminal peptides of collagen alpha (XV), etc. Cytotoxic and
cytostatic agents of interest include adriamycin, aleran, Ara-C,
BICNU, busulfan, CNNU, cisplatinum, cytoxan, daunorubicin, DTIC,
5-FU, hydrea, ifosfamicle, methotrexate, mithramycin, mitomycin,
mitoxantrone, nitrogen mustard, velban, vincristine, vinblastine,
VP-16, carboplatinum, fludarabine, gemcitabine, idarubicin,
irinotecan, leustatin, navelbine, taxol, taxotere, topotecan, etc.
For example, in the case of bone conditions, combinations that
would be favourable include those with antiresorptive
bisphosphonates, such as alendronate and risedronate; integrin
blockers (as defined further below), such as
.alpha..sub.v.beta..sub.3 antagonists; conjugated oestrogens used
in hormone replacement therapy, such as PREMPRO.RTM., PREMARIN.RTM.
and ENDOMETRION.RTM.; selective oestrogen receptor modulators
(SERMs), such as raloxifene, droloxifene, CP-336,156 (Pfizer) and
lasofoxifene; cathepsin K inhibitors; and ATP proton pump
inhibitors.
[0232] The present compounds are also suitable for combination with
known anti-cancer agents. These known anti-cancer agents include
the following: oestrogen receptor modulators androgen receptor
modulators, retinoid receptor modulators, cytotoxic agents,
antiproliferative agents, prenyl-protein transferase inhibitors,
HMG-CoA reductase inhibitors, HIV protease inhibitors, reverse
transcriptase inhibitors and other angiogenesis inhibitors. The
present compounds are particularly suitable for administration at
the same time as radiotherapy. The synergistic effects of
inhibiting VEGF in combination with radiotherapy have been
described in the art (see WO00/61186).
[0233] Therefore, a preferred embodiment of the present invention
is the use of a compound of the general formula I for the
manufacture of a medicament for the treatment and/or prevention of
disorders, characterized in that a therapeutically effective amount
of one or more compounds according to the present invention is
administered in combination with an compound selected from the
group consisting of estrogen receptor modulators, androgen receptor
modulators, retinoid receptor modulators, cytotoxic agents,
anti-proliferative agents, prenyl protein protease inhibitors, HMG
CoA reductase inhibitors, HIV protease inhibitors, reverse
transcriptase inhibitors, growth factor receptor inhibitors and
angiogenesis inhibitors.
[0234] Additionally, a preferred embodiment of the present
invention is the use of a compound of the general formula I for the
manufacture of a medicament for the treatment and/or prevention of
disorders, characterized in that a therapeutically effective amount
of one or more compounds according to the present invention is
administered in combination radio therapy and with an compound
selected from the group consisting of estrogen receptor modulators,
androgen receptor modulators, retinoid receptor modulators,
cytotoxic agents, anti-proliferative agents, prenyl protein
protease inhibitors, HMG CoA reductase inhibitors, HIV protease
inhibitors, reverse transcriptase inhibitors, growth factor
receptor inhibitors and angiogenesis inhibitors.
[0235] "Oestrogen receptor modulators" refers to compounds, which
interfere with or inhibit the binding of oestrogen to the receptor,
regardless of mechanism. Examples of oestrogen receptor modulators
include, for example, tamoxifen, raloxifene, idoxifene, LY353381,
LY 117081, toremifene, fulvestrant,
4-[7-(2,2-dimethyl-1-oxopropoxy-4-methyl-2-[4-[2-(1-piperidinyl)ethoxy]ph-
enyl]-2H-1-benzopyran-3-yl]phenyl-2,2-dimethylpropanoate,
4,4''-dihydroxybenzophenone-2,4-dinitrophenylhydrazone and SH646.
"Androgen receptor modulators" refers to compounds, which interfere
with or inhibit the binding of androgens to the receptor,
regardless of mechanism. Examples of androgen receptor modulators
include finasteride and other 5.alpha.-reductase inhibitors,
nilutamide, flutamide, bicalutamide, liarozole and abiraterone
acetate.
[0236] "Retinoid receptor modulators" refers to compounds, which
interfere with or inhibit the binding of retinoids to the receptor,
regardless of mechanism. Examples of such retinoid receptor
modulators include bexarotene, tretinoin, 13-cis-retinoic acid,
9-cis-retinoic acid, .alpha.-difluoromethylornithine, ILX23-7553,
trans-N-(4'-hydroxyphenyl)retinamide and N-4-carboxyphenyl
retinamide.
[0237] "Cytotoxic agents" refers to compounds, which result in cell
death primarily through direct action on the cellular function or
inhibit or interfere with cell myosis, including alkylating agents,
tumour necrosis factors, intercalators, microtubulin inhibitors and
topoisomerase inhibitors.
[0238] Examples of cytotoxic agents include, but are not limited
to, tirapazimine, sertenef, cachectin, ifosfamide, tasonermin,
lonidamine, carboplatin, altretamine, prednimustine,
dibromodulcitol, ranimustine, fotemustine, nedaplatin, oxaliplatin,
temozolomide, heptaplatin, estramustine, improsulfan tosylate,
trofosfamide, nimustine, dibrospidium chloride, pumitepa,
lobaplatin, satraplatin, profiromycin, cisplatin, irofulven,
dexifosfamide, cis-aminedichloro(2-methylpyridine) platinum,
benzylguanine, glufosfamide, GPX100,
(trans,trans,trans)-bis-mu-(hexane-1,6-diamine)-mu-[diamine-platinum(II)]-
bis[diamine(chloro)platinum (II)]tetrachloride,
diarizidinyl-spermine, arsenic trioxide,
1-(11-dodecylamino-10-hydroxyundecyl)-3,7-dimethylxanthine,
zorubicin, idarubicin, daunorubicin, bisantrene, mitoxan-trone,
pirarubicin, pinafide, valrubicin, amrubicin, antineoplaston,
3'-deamino-3'-morpholino-13-deoxo-10-hydroxycaminomycin, annamycin,
galarubicin, elinafide, MEN10755 and
4-demethoxy-3-deamino-3-aziridinyl-4-methylsulfonyldaunorubicin
(see WO 00/50032).
[0239] Examples of microtubulin inhibitors include paclitaxel,
vindesine sulfate,
3',4'-didehydro-4'-deoxy-8'-norvincaleukoblastine, docetaxol,
rhizoxin, dolastatin, mivobulin isethionate, auristatin, cemadotin,
RPR109881, BMS184476, vinflunine, cryptophycin,
2,3,4,5,6-pentafluoro-N-(3-fluoro-4-methoxyphenyl)benzenesulfonamide,
anhydrovinblastine,
N,N-dimethyl-L-valyl-L-valyl-N-methyl-L-valyl-L-prolyl-L-proline-t-butyla-
mide, TDX258 and BMS188797.
[0240] Some examples of topoisomerase inhibitors are topotecan,
hycaptamine, irinotecan, rubitecan,
6-ethoxypropionyl-3',4'-O-exobenzylidene-chartreusin,
9-methoxy-N,N-dimethyl-5-nitropyrazolo[3,4,5-kl]acridine-2-(6H)propanamin-
e,
1-amino-9-ethyl-5-fluoro-2,3-dihydro-9-hydroxy-4-methyl-1H,12H-benzo[de-
]pyrano[3',4':b,7]indolizino[1,2b]quinoline-10,13(9H,15H)dione,
lurtotecan, 742-(N-isopropylamino)ethyl]-(20S)-camptothecin,
BNP1350, BNPI1100, BN80915, BN80942, etoposide phosphate,
teniposide, sobuzoxane, 2'-dimethylamino-2'-deoxyetoposide, GL331,
N42-(dimethylamino)ethyl]-9-hydroxy-5,6-dimethyl-6H-pyrido[4,3-b]carbazol-
e-1-carboxamide, asulacrine,
(5a,5aB,8aa,9b)-9-[2-[N-[2-(dimethylamino)ethyl]-methylamino]ethyl]-5-[4--
hydroxy-3,5-dimethoxy-phenyl]-5,5a,6,8,8a,9-hexohydrofuro(3',4':6,7)naphth-
o(2,3-d)-1,3-dioxol-6-one,
2,3-(methylenedioxy)-5-methyl-7-hydroxy-8-methoxybenzo[c]phen-anthridiniu-
m, 6,9-bis[(2-aminoethyl)amino]benzo[g]isoquinoline-5,10-dione,
5-(3-aminopropylamino)-7,10-dihydroxy-2-(2-hydroxyethylaminomethyl)-6H-py-
razolo[4,5,1-de]acridin-6-one,
N-[1-[2(diethylamino)ethylamino]-7-methoxy-9-oxo-9H-thioxanthen-4-ylmethy-
l]formamide, N-(2-(dimethylamino)ethyl)acridine-4-carboxamide,
6-[[2-(dimethylamino)ethyl]-amino]-3-hydroxy-7H-indeno[2,1-c]quinolin-7-o-
ne and dimesna. "Antiproliferative agents" include antisense RNA
and DNA oligonucleotides such as G3139, ODN698, RVASKRAS, GEM231
and INX3001 and antimetabolites such as enocitabine, carmofur,
tegafur, pentostatin, doxifluridine, trimetrexate, fludarabine,
capecitabine, galocitabine, cytarabine ocfosfate, fosteabine sodium
hydrate, raltitrexed, paltitrexid, emitefur, tiazofurin,
decitabine, nolatrexed, pemetrexed, nelzarabine,
2'-deoxy-2'-methylidenecytidine,
2'-fluoromethylene-2'-deoxycytidine,
N-[5-(2,3-dihydrobenzofuryl)sulfonyl]-N'-(3,4-dichlorophenyl)urea,
N6-[4-deoxy-4-[N2-[2(E),4(E)-tetradecadienoyl]glycylamino]-L-glycero-B-L--
manno-heptopyranosyl]adenine, aplidine, ecteinascidin,
troxacitabine,
4-[2-amino-4-oxo-4,6,7,8-tetrahydro-3H-pyrimidino[5,4-b][1,4]thiazin-6-yl-
-(S)-ethyl]-2,5-thienoyl-L-glutamic acid, aminopterin,
5-fluorouracil, alanosine,
11-acetyl-8-(carbamoyloxymethyl)-4-formyl-6-methoxy-[4-oxa-1,1'-diaza-tet-
racyclo(7.4.1.0.0)tetradeca-2,4,6-trien-9-ylacetic acid ester,
swainsonine, lometrexol, dexrazoxane, methioninase,
2'-cyano-2'-deoxy-N4-palmitoyl-1-B-D-arabinofuranosyl cytosine and
3-aminopyridine-2-carboxaldehyde thiosemicarbazone.
"Antiproliferative agents" also include monoclonal antibodies to
growth factors other than those listed under "angiogenesis
inhibitors", such as trastuzumab and tumour suppressor genes, such
as p53, which can be delivered via recombinant virus-mediated gene
transfer (see U.S. Pat. No. 6,069,134, for example).
[0241] As stated above the compounds according of the formula I are
effective modulators (activators or inhibitors) of one or more
protein kinases selected from the group of Raf, Mek, PKB, Tie2,
PDGFR and VEGFR.
[0242] Therefore, a preferred embodiment of the present invention
are compounds of formula I as protein kinase activators or
inhibitors.
[0243] A particularly preferred embodiment of the present invention
are compounds of formula I as protein kinase activators or
inhibitors, characterized in that the protein kinase is MEK1 or
MEK2.
[0244] Thus, the compounds of the invention may also be useful as
reagents for studying signal transduction, protein kinases or any
of the clinical disorders listed throughout this application.
[0245] For the identification of a signal transduction pathway and
the detection of cross talks with other signaling pathways suitable
models or model systems have been generated by various scientists,
for example cell culture models (e.g. Khwaja et al., EMBO, 1997,
16, 2783-93) and transgenic animal models (e.g. White et al.,
Oncogene, 2001, 20, 7064-7072). For the examination of particular
steps in the signal transduction cascade, interfering compounds can
be used for signal modulation (e.g. Stephens et al., Biochemical
J., 2000, 351, 95-105). The compounds according to the invention
may also be useful as reagents for the examination of kinase
dependent signal transduction pathways in animal and/or cell
culture models or any of the clinical disorders listed throughout
this application.
[0246] The measurement of kinase activity is a well-known technique
feasible for each person skilled in the art. Generic test systems
for kinase activity detection with substrates, for example histone
(e.g. Alessi et al., FEBS Lett. 1996, 399, 3, page 333-8) or myelin
basic protein are well described in the literature (e.g.
Campos-Gonzalez, R. and Glenney, Jr., J. R. 1992 J. Biol. Chem.
267, Page 14535).
[0247] For the identification of kinase inhibitors various assay
systems are available (see for example Walters et al., Nature Drug
Discovery 2003, 2; page 259-266). For example, in scintillation
proximity assays (e.g. Sorg et al., J. of. Biomolecular Screening,
2002, 7, 11-19) or flashplate assays the radioactive
phosphorylation of a protein or peptide as substrate with
.gamma.ATP can be measured. In the presence of an inhibitory
compound no signal or a decreased radioactive signal is detectable.
Furthermore homogeneous time-resolved fluorescence resonance energy
transfer (HTR-FRET), and fluorescence polarization (FP)
technologies are useful for assay methods (for example Sills et
al., J. of Biomolecular Screening, 2002: 191-214).
[0248] Other non-radioactive ELISA based assay methods use specific
phospho-antibodies (AB). The phospho-AB binds only the
phosphorylated substrate. This binding is detectable with a
secondary peroxidase conjugated antibody, measured for example by
chemiluminescence (for example Ross et al., Biochem. J., 2002, 366:
977-981).
[0249] Other assays are known from the literature and could readily
be performed by the person skilled in the art (see, for example,
Dhanabal et al., Cancer Res. 59:189-197; Xin et al., J. Biol. Chem.
274:9116-9121; Sheu et al., Anticancer Res. 18:4435-4441; Ausprunk
et al., Dev. Biol. 38:237-248; Gimbrone et al., J. Natl. Cancer
Inst. 52:413-427; Nicosia et al., In Vitro 18:538-549).
[0250] A further preferred embodiment of the present invention is a
pharmaceutical composition, characterized in that it contains a
therapeutically effective amount of one or more compounds according
to the invention.
[0251] A further embodiment of the present invention is a
pharmaceutical composition, characterized in that it further
contains one or more additional compounds, selected from the group
consisting of physiologically acceptable excipients, auxiliaries,
adjuvants, diluents, carriers and pharmaceutically active agents
other than the compounds according to the invention.
[0252] An additional preferred embodiment of the present invention
is a set (kit) consisting of separate packets of [0253] a) a
therapeutically effective amount of one or more compounds according
to the invention and [0254] b) a therapeutically effective amount
one or more further pharmaceutically active agents other than the
compounds according to the invention.
[0255] A further embodiment of the present invention is a process
for the manufacture of said pharmaceutical compositions,
characterized in that one or more compounds according to the
invention and one or more compounds selected from the group
consisting of solid, liquid or semiliquid excipients, auxiliaries,
adjuvants, diluents, carriers and pharmaceutically active agents
other than the compounds according to the invention, are converted
in a suitable dosage form.
[0256] The pharmaceutical compositions of the present invention may
be administered by any means that achieve their intended purpose.
For example, administration may be by oral, parenteral, topical,
enteral, intravenous, intramuscular, inhalant, nasal,
intraarticular, intraspinal, transtracheal, transocular,
subcutaneous, intraperitoneal, transdermal, or buccal routes.
Alternatively, or concurrently, administration may be by the oral
route. The dosage administered will be dependent upon the age,
health, and weight of the recipient, kind of concurrent treatment,
if any, frequency of treatment, and the nature of the effect
desired. Parenteral administration is preferred. Oral
administration is especially preferred.
[0257] Suitable dosage forms include, but are not limited to
capsules, tablets, pellets, dragees, semi-solids, powders,
granules, suppositories, ointments, creams, lotions, inhalants,
injections, cataplasms, gels, tapes, eye drops, solution, syrups,
aerosols, suspension, emulsion, which can be produced according to
methods known in the art, for example as described below:
Tablets:
[0258] mixing of active ingredients and auxiliaries, compression of
said mixture into tablets (direct compression), optionally
granulation of part of mixture before compression.
Capsules:
[0259] mixing of active ingredient/s and auxiliaries to obtain a
flowable powder, optionally granulating powder, filling
powders/granulate into opened capsules, capping of capsules.
Semi-Solids (Ointments, Gels, Creams):
[0260] dissolving/dispersing active ingredient/s in an aqueous or
fatty carrier; subsequent mixing of aqueous/fatty phase with
complementary fatty resp. aqueous phase, homogenization (creams
only).
Suppositories (Rectal and Vaginal):
[0261] dissolving/dispersing active ingredient/s in carrier
material liquified by heat (rectal: carrier material normally a
wax; vaginal: carrier normally a heated solution of a gelling
agent), casting said mixture into suppository forms, annealing and
withdrawal suppositories from the forms.
Aerosols:
[0262] dispersing/dissolving active agents in a propellant,
bottling said mixture into an atomizer.
[0263] In general, non-chemical routes for the production of
pharmaceutical compositions and/or pharmaceutical preparations
comprise processing steps on suitable mechanical means known in the
art that transfer one or more compounds according to the invention
into a dosage form suitable for administration to a patient in need
of such a treatment. Usually, the transfer of one or more compounds
according to the invention into such a dosage form comprises the
addition of one or more compounds, selected from the group
consisting of carriers, excipients, auxiliaries and pharmaceutical
active ingredients other than the compounds according to the
invention. Suitable processing steps include, but are not limited
to combining, milling, mixing, granulating, dissolving, dispersing,
homogenizing, casting and/or compressing the respective active and
non-active ingredients. Mechanical means for performing said
processing steps are known in the art, for example from Ullmann's
Encyclopedia of Industrial Chemistry, 5th Edition. In this respect,
active ingredients are preferably at least one compound according
to this invention and one or more additional compounds other than
the compounds according to the invention, which show valuable
pharmaceutical properties, preferably those pharmaceutical active
agents other than the compounds according to the invention, which
are disclosed herein.
[0264] Particularly suitable for oral use are tablets, pills,
coated tablets, capsules, powders, granules, syrups, juices or
drops, suitable for rectal use are suppositories, suitable for
parenteral use are solutions, preferably oil-based or aqueous
solutions, furthermore suspensions, emulsions or implants, and
suitable for topical use are ointments, creams or powders. The
novel compounds may also be lyophilised and the resultant
lyophilisates used, for example, for the preparation of injection
preparations. The preparations indicated may be sterilised and/or
comprise assistants, such as lubricants, preservatives, stabilisers
and/or wetting agents, emulsifiers, salts for modifying the osmotic
pressure, buffer substances, dyes, flavours and/or a plurality of
further active ingredients, for example one or more vitamins.
[0265] Suitable excipients are organic or inorganic substances,
which are suitable for enteral (for example oral), parenteral or
topical administration and do not react with the novel compounds,
for example water, vegetable oils, benzyl alcohols, alkylene
glycols, polyethylene glycols, glycerol triacetate, gelatine,
carbohydrates, such as lactose, sucrose, mannitol, sorbitol or
starch (maize starch, wheat starch, rice starch, potato starch),
cellulose preparations and/or calcium phosphates, for example
tricalcium phosphate or calcium hydrogen phosphate, magnesium
stearate, talc, gelatine, tragacanth, methyl cellulose,
hydroxypropylmethylcellulose, sodium carboxymethylcellulose,
polyvinyl pyrrolidone and/or Vaseline.
[0266] If desired, disintegrating agents may be added such as the
above-mentioned starches and also carboxymethyl-starch,
cross-linked polyvinyl pyrrolidone, agar, or alginic acid or a salt
thereof, such as sodium alginate. Auxiliaries include, without
limitation, flow-regulating agents and lubricants, for example,
silica, talc, stearic acid or salts thereof, such as magnesium
stearate or calcium stearate, and/or polyethylene glycol. Dragee
cores are provided with suitable coatings, which, if desired, are
resistant to gastric juices. For this purpose, concentrated
saccharide solutions may be used, which may optionally contain gum
arabic, talc, polyvinyl pyrrolidone, polyethylene glycol and/or
titanium dioxide, lacquer solutions and suitable organic solvents
or solvent mixtures. In order to produce coatings resistant to
gastric juices or to provide a dosage form affording the advantage
of prolonged action, the tablet, dragee or pill can comprise an
inner dosage and an outer dosage component me latter being in the
form of an envelope over the former. The two components can be
separated by an enteric layer, which serves to resist
disintegration in the stomach and permits the inner component to
pass intact into the duodenum or to be delayed in release. A
variety of materials can be used for such enteric layers or
coatings, such materials including a number of polymeric acids and
mixtures of polymeric acids with such materials as shellac, acetyl
alcohol, solutions of suitable cellulose preparations such as
acetyl-cellulose phthalate, cellulose acetate or
hydroxypropymethyl-cellulose phthalate, are used. Dye stuffs or
pigments may be added to the tablets or dragee coatings, for
example, for identification or in order to characterize
combinations of active compound doses.
[0267] Suitable carrier substances are organic or inorganic
substances which are suitable for enteral (e.g. oral) or parenteral
administration or topical application and do not react with the
novel compounds, for example water, vegetable oils, benzyl
alcohols, polyethylene glycols, gelatin, carbohydrates such as
lactose or starch, magnesium stearate, talc and petroleum jelly. In
particular, tablets, coated tablets, capsules, syrups, suspensions,
drops or suppositories are used for enteral administration,
solutions, preferably oily or aqueous solutions, furthermore
suspensions, emulsions or implants, are used for parenteral
administration, and ointments, creams or powders are used for
topical application. The novel compounds can also be lyophilized
and the lyophilizates obtained can be used, for example, for the
production of injection preparations.
[0268] The preparations indicated can be sterilized and/or can
contain excipients such as lubricants, preservatives, stabilizers
and/or wetting agents, emulsifiers, salts for affecting the osmotic
pressure, buffer substances, colorants, flavourings and/or
aromatizers. They can, if desired, also contain one or more further
active compounds, e.g. one or more vitamins.
[0269] Other pharmaceutical preparations, which can be used orally
include push-fit capsules made of gelatine, as well as soft, sealed
capsules made of gelatine and a plasticizer such as glycerol or
sorbitol. The push-fit capsules can contain the active compounds in
the form of granules, which may be mixed with fillers such as
lactose, binders such as starches, and/or lubricants such as talc
or magnesium stearate and, optionally, stabilizers. In soft
capsules, the active compounds are preferably dissolved or
suspended in suitable liquids, such as fatty oils, or liquid
paraffin. In addition, stabilizers may be added.
[0270] The liquid forms in which the novel compositions of the
present invention may be incorporated for administration orally
include aqueous solutions, suitably flavoured syrups, aqueous or
oil suspensions, and flavoured emulsions with edible oils such as
cottonseed oil, sesame oil, coconut oil or peanut oil, as well as
elixirs and similar pharmaceutical vehicles. Suitable dispersing or
suspending agents for aqueous suspensions include synthetic and
natural gums such as tragacanth, acacia, alginate, dextran, sodium
carboxymethylcellulose, methylcellulose, polyvinyl-pyrrolidone or
gelatine.
[0271] Suitable formulations for parenteral administration include
aqueous solutions of the active compounds in water-soluble form,
for example, water-soluble salts and alkaline solutions. In
addition, suspensions of the active compounds as appropriate oily
injection suspensions may be administered. Suitable lipophilic
solvents or vehicles include fatty oils, for example, sesame oil,
or synthetic fatty acid esters, for example, ethyl oleate or
triglycerides or polyethylene glycol-400 (the compounds are soluble
in PEG-400).
[0272] Aqueous injection suspensions may contain substances, which
increase the viscosity of the suspension include, for example,
sodium carboxymethyl cellulose, sorbitol, and/or dextran,
optionally, the suspension may also contain stabilizers.
[0273] For administration as an inhalation spray, it is possible to
use sprays in which the active ingredient is either dissolved or
suspended in a propellant gas or propellant gas mixture (for
example CO.sub.2 or chlorofluorocarbons). The active ingredient is
advantageously used here in micronized form, in which case one or
more additional physiologically acceptable solvents may be present,
for example ethanol. Inhalation solutions can be administered with
the aid of conventional inhalers.
[0274] Possible pharmaceutical preparations, which can be used
rectally include, for example, suppositories, which consist of a
combination of one or more of the active compounds with a
suppository base. Suitable suppository bases are, for example,
natural or synthetic triglycerides, or paraffin hydrocarbons. In
addition, it is also possible to use gelatine rectal capsules,
which consist of a combination of the active compounds with a base.
Possible base materials include, for example, liquid triglycerides,
polyethylene glycols, or paraffin hydrocarbons.
[0275] For use in medicine, the compounds of the present invention
will be in the form of pharmaceutically acceptable salts. Other
salts may, however, be useful in the preparation of the compounds
according to the invention or of their pharmaceutically acceptable
salts. Suitable pharmaceutically acceptable salts of the compounds
of this invention include acid addition salts which may, for
example be formed by mixing a solution of the compound according to
the invention with a solution of a pharmaceutically acceptable acid
such as hydrochloric acid, sulphuric acid, methanesulphonic acid,
fumaric acid, maleic acid, succinic acid, acetic acid, benzoic
acid, oxalic acid, citric acid, tartaric acid, carbonic acid or
phosphoric acid. Furthermore, where the compounds of the invention
carry an acidic moiety, suitable pharmaceutically acceptable salts
thereof may include alkali metal salts, e.g. sodium or potassium
salts; alkaline earth metal salts, e.g. calcium or magnesium salts;
and salts formed with suitable organic bases, e.g. quaternary
ammonium salts.
[0276] The present invention includes within its scope prodrugs of
the compounds of the present invention above. In general, such
prodrugs will be functional derivatives of the compounds of the
present invention, which are readily convertible in vivo into the
required compound of the present invention. Conventional procedures
for the selection and preparation of suitable prodrug derivatives
are described, for example, in Design of Prodrugs, ed. H.
Bundgaard, Elsevier, 1985.
[0277] The pharmaceutical preparations can be employed as
medicaments in human and veterinary medicine. As used herein, the
term "effective amount" means that amount of a drug or
pharmaceutical agent that will elicit the biological or medical
response of a tissue, system, animal or human that is being sought,
for instance, by a researcher or clinician. Furthermore, the term
"therapeutically effective amount" means any amount which, as
compared to a corresponding subject who has not received such
amount, results in improved treatment, healing, prevention, or
amelioration of a disease, disorder, or side effect, or a decrease
in the rate of advancement of a disease or disorder. The term also
includes within its scope amounts effective to enhance normal
physiological function. Said therapeutic effective amount of one or
more of the compounds according to the invention is known to the
skilled artisan or can be easily determined by standard methods
known in the art.
[0278] The substances according to the invention are generally
administered analogously to commercial preparations. Usually,
suitable doses that are therapeutically effective lie in the range
between 0.0005 mg and 1000 mg, preferably between 0.005 mg and 500
mg and especially between 0.5 and 100 mg per dose unit. The daily
dose is preferably between about 0.001 and 10 mg/kg of body
weight.
[0279] Those of skill will readily appreciate that dose levels can
vary as a function of the specific compound, the severity of the
symptoms and the susceptibility of the subject to side effects.
Some of the specific compounds are more potent than others.
Preferred dosages for a given compound are readily determinable by
those of skill in the art by a variety of means. A preferred means
is to measure the physiological potency of a given compound.
[0280] The host, or patient, may be from any mammalian species,
e.g., primate sp., particularly human; rodents, including mice,
rats and hamsters; rabbits; equines, bovines, canines, felines;
etc. Animal models are of interest for experimental investigations,
providing a model for treatment of human disease.
[0281] The specific dose for the individual patient depends,
however, on the multitude of factors, for example on the efficacy
of the specific compounds employed, on the age, body weight,
general state of health, the sex, the kind of diet, on the time and
route of administration, on the excretion rate, the kind of
administration and the dosage form to be administered, the
pharmaceutical combination and severity of the particular disorder
to which the therapy relates. The specific therapeutic effective
dose for the individual patient can readily be determined by
routine experimentation, for example by the doctor or physician,
which advises or attends the therapeutic treatment.
[0282] In the case of hyperproliferative disorders, the
susceptibility of a particular cell to treatment with the subject
compounds may be determined by in vitro testing. Typically a
culture of the cell is combined with a subject compound at varying
concentrations for a period of time sufficient to allow the active
agents to induce cell death or inhibit migration, usually between
about one hour and one week. For in vitro testing, cultured cells
from a biopsy sample may be used. The viable cells, left after
treatment, are then counted.
[0283] The dose will vary depending on the specific compound
utilized, specific disorder, patient status, etc. Typically a
therapeutic dose will be sufficient to substantially decrease the
undesirable cell population in the targeted tissue, while
maintaining patient viability. Treatment will generally be
continued until there is a substantial reduction, e.g., at least
about 50%, decrease in the cell burden, and may be continued until
there are essentially none of the undesirable cells detected in the
body.
[0284] Even without further details, it is assumed that a person
skilled in the art will be able to utilise the above description in
the broadest scope. The preferred embodiments should therefore
merely be regarded as descriptive disclosure, which is absolutely
not limiting in any way.
[0285] Above and below, all temperatures are indicated in .degree.
C. In the following examples, "conventional work-up" means that, if
necessary, the solvent is removed, water is added if necessary, the
pH is adjusted, if necessary, to between 2 and 10, depending on the
constitution of the end product, the mixture is extracted with
ethyl acetate or dichloromethane, the phases are separated, the
organic phase is washed with saturated NaHCO.sub.3 solution, if
desired with water and saturated NaCl solution, is dried over
sodium sulfate, filtered and evaporated, and the product is
purified by chromatography on silica gel, by preparative HPLC
and/or by crystallisation. The purified compounds are, if desired,
freeze-dried.
[0286] Mass spectrometry (MS): ESI (electrospray ionisation)
(M+H).sup.+
List of Abbreviations and Acronyms:
[0287] AcOH acetic acid, anh anhydrous, atm atmosphere(s), BOC
tert-butoxycarbonyl CDI 1,1'-carbonyl diimidazole, cone
concentrated, d day(s), dec decomposition, DMAC
N,N-dimethylacetamide, DMPU
1,3-dimethyl-3,4,5,6-tetrahydro-2(IH)-pyrimidinone, DMF
N,N-dimethylformamide, DMSO dimethylsulfoxide, DPPA
diphenylphosphoryl azide, EDCl
1-(3-dimethylaminopropyl)-3-ethylcarbodiimide, EtOAc ethyl acetate,
EtOH ethanol (100%), Et.sub.2Q diethyl ether, Et.sub.3N
triethylamine, h hour(s), MeOH methanol, pet. ether petroleum ether
(boiling range 30-60.degree. C.), temp. temperature, THF
tetrahydrofuran, TFA trifluoroAcOH, Tf
trifluoromethanesulfonyl.
EXAMPLE 1
Preparation of Amino-Oxindoles
##STR00006##
[0289] One equivalent of a substituted aromatic or heteroaromatic
nitrile (here a compound of formula II, wherein R.sup.1 is phenyl,
R.sup.7 is I, X is (CH.sub.2).sub.p and p=0) is dissolved in
1,4-dioxane. After addition of 1,3 equivalents of a straight or
branched chain alcohol of formula III (wherein R.sup.9 is
(CH.sub.2).sub.q and q is 1-10), HCl-gas is introduced at
10.degree. C. for 6 h. The resulting pellet (compound of formula
IV) is filtered, washed with dioxane and dried in vacuum.
[0290] IVa) 3-Iodo-benzimidic acid ethyl ester hydrochloride: 6.05
g (81%), colourless powder; HPLC: 2.11 min; LC-MS: 262.0 m/z.
[0291] IVb) 4-Iodo-benzimidic acid ethyl ester hydrochloride: 4.80
g (62%), beige crystals; HPLC: 2.00 min; LC-MS: 262.0 m/z.
[0292] Triethylamine (3 eq.), the compound of formula IV (2 eq.)
and the compound of formula V (1 eq.) are dissolved in 1-butanol (7
eq.) and are irradiated for 70 min at 200.degree. C. in a
microwave. The resulting product (respective compound of formula I,
see following table) is purified by column chromatography.
TABLE-US-00001 Retention time Amount/ Yield/ HPLC/ LC-MS/ No.
Structure mg % min mz-1 IC.sub.50 a ##STR00007## 37 22 2.77 271.0
1.10E-06 b ##STR00008## 47 29 2.75 271.0 2.60E-07 c ##STR00009## 14
7 2.33 253.0 9.40E-07 d ##STR00010## 16 6 2.83 363.0 2.70E-06 e
##STR00011## 37 15 3.00 397.0 4.10E-06 f ##STR00012## 19 8 2.95
397.0 1.10E-06 g ##STR00013## 44 16 2.79 363.0 n.d. h ##STR00014##
30 13 2.95 397.0 n.d. i ##STR00015## 44 19 2.91 397.0 n.d. j
##STR00016## 43 17 2.97 377.0 n.d. k ##STR00017## 33 22 2.77 316.0
6.60E-07 l ##STR00018## 25 10 2.95 377.0 n.d. m ##STR00019## 24 11
2.95 440.9 n.d. n ##STR00020## 20 10 2.95 440.8 3.40E-06 o
##STR00021## 21 10 2.59 267.2 3.10E-06 p ##STR00022## 37 20 2.72
301.0 8.70E-06 q ##STR00023## 25 15 2.73 345.0 n.d. r ##STR00024##
98 19 2.49 287.0 n.d. s ##STR00025## 260 51 2.47 287.0 n.d. t
##STR00026## 59 12 2.71 251.2 n.d. u ##STR00027## 70 13 2.48 267.2
n.d. v ##STR00028## 64 14 2.52 331.0 n.d. w ##STR00029## 97 18 2.67
301.0 n.d. x ##STR00030## 47 12 2.6 255.2 n.d. y ##STR00031## 51 15
2.73 289.0 n.d. z ##STR00032## 31 8 2.67 281.2 n.d.
EXAMPLE 2
Cellular Assay for Measuring MEK Inhibition
[0293] MEK inhibitors are evaluated by determining their ability to
inhibit phosphorylation of MAP kinase (ERK) in murine colon 26
(C26) carcinoma cells. Since ERK1 and ERK2 represent the only known
substrates for MEK1 and MEK2, the measurement of inhibition of ERK
phosphorylation in cells provides direct read out of cellular MEK
inhibition by the compounds of the invention. Detection of
phosphorylation of ERK is carried out either by Western blot or
ELISA format. Briefly, the assays involve treatment of
exponentially growing C26 cells with varying concentrations of the
test compound (or vehicle control) for one hour at 37.degree. C.
For Western blot assay, cells are rinsed free of compound/vehicle
and lysed in a solution containing 70 mM NaCl, 50 mM glycerol
phosphate, 10 mM HEPES, pH 7.4, 1% Triton X-100, 1 mM
Na.sub.3VO.sub.4, 100 .mu.M PMSF, 10 .mu.M leupeptin and 10 .mu.M
pepstatin. Supernatants were then subjected to gel electrophoresis
and hybridized to a primary antibody recognizing dually
phosphorylated ERKl and ERK2. To evaluate total MAPK levels, blots
were subsequently `stripped` and re-probed with a 1:1 mixture of
polyclonal antibodies recognizing unphosphorylated ERK1 and ERK2.
For pERK ELISA assay, pERK TiterZyme Enzyme immunometric Assay kits
were acquired from Assay Designs, Inc (Ann Arbor, Mich.). Briefly,
cells were harvested in lysis solution containing 50 mM
B-glycerophosphate, 10 mM HEPES, pH 7.4, 70 mM NaCl, 2 mM EDTA and
1% SDS and protein lysates were diluted 1:15 with supplied Assay
buffer prior to the execution of the assay. The subsequent steps
were carried out essentially as recommended by the
manufacturer.
EXAMPLE 3
VEGF Receptor Kinase Assay
[0294] VEGF receptor kinase activity is measured by incorporation
of radio-labelled phosphate into polyglutamic acid/tyrosine, 4:1
(pEY) substrate. The phosphorylated pEY product is trapped onto a
filter membrane and the incorporation of radiolabelled phosphate is
quantified by scintillation counting.
Materials
[0295] VEGF receptor kinase: The intracellular tyrosine kinase
domains of human KDR (Terman, B. I. et al. Oncogene (1991) Vol. 6,
pp. 1677-1683.) and Flt-1 (Shibuya, M. et al. Oncogene (1990) Vol.
5, pp. 519-524) were cloned as glutathione S-transferase (GST) gene
fusion proteins. This was accomplished by cloning the cytoplasmic
domain of the KDR kinase as an in frame fusion at the carboxyl
terminus of the GST gene. Soluble recombinant GST-kinase domain
fusion proteins are expressed in Spodoptera frugiperda (Sf21)
insect cells (Invitrogen) using a baculovirus expression vector
(pAcG2T, Pharmingen).
[0296] Lysis buffer: 50 mM Tris pH 7.4, 0.5 M NaCl, 5 mM DTT, 1 mM
EDTA, 0.5% triton X-100, 10% glycerol, 10 mg/ml of each leupeptin,
pepstatin and aprotinin and 1 mM phenylmethylsulfonyl fluoride (all
Sigma).
[0297] Wash buffer: 50 mM Tris pH 7.4, 0.5 M NaCl, 5 mM DTT, 1 mM
EDTA, 0.05% triton X-100, 10% glycerol, 10 mg/ml of each leupeptin,
pepstatin and aprotinin and 1 mM phenylmethylsulfonyl fluoride.
[0298] Dialysis buffer: 50 mM Tris pH 7.4, 0.5 M NaCl, 5 mM DTT, 1
mM EDTA, 0.05% triton X-100, 50% glycerol, 10 mg/ml of each
leupeptin, pepstatin and aprotinin and 1 mM phenylmethylsulfonyl
fluoride.
[0299] 10.times. reaction buffer: 200 mM Tris, pH 7.4, 1.0 M NaCl,
50 mM MnCl.sub.2, 10 mM DTT and 5 mg/ml bovine serum albumin [BSA]
(Sigma).
[0300] Enzyme dilution buffer: 50 mM Tris, pH 7.4, 0.1 M NaCl, 1 mM
DTT, 10% glycerol, 100 mg/ml BSA.
[0301] 10.times. substrate: 750 .mu.g/ml poly(glutamic
acid/tyrosine; 4:1) (Sigma).
[0302] Stop solution: 30% trichloroacetic acid, 0.2 M sodium
pyrophosphate (both Fisher).
[0303] Wash solution: 15% trichloroacetic acid, 0.2 M sodium
pyrophosphate.
Filter Plates
[0304] Millipore #MAFC NOB, GF/C glass fibre 96 well plate.
Method A--Protein Purification
[0305] 1. Sf21 cells are infected with recombinant virus at a
multiplicity of infection of 5 virus particles/cell and grown at
27.degree. C. for 48 hours.
[0306] 2. All steps are performed at 4.degree. C. Infected cells
are harvested by centrifugation at 1000.times.g and lysed at
4.degree. C. for 30 minutes with 1/10 volume of lysis buffer
followed by centrifugation at 100,000.times.g for 1 hour. The
supernatant is then passed over a glutathione Sepharose column
(Pharmacia) equilibrated with lysis buffer and washed with 5
volumes of the same buffer followed by 5 volumes of wash buffer.
Recombinant GST-KDR protein is eluted with wash buffer/10 mM
reduced glutathione (Sigma) and dialysed against dialysis
buffer.
Method B--VEGF Receptor Kinase Assay
[0307] 1, Add 5 .mu.l of inhibitor or control to the assay in 50%
DMSO.
[0308] 2. Add 35 .mu.l of reaction mixture containing 5 .mu.l of
10.times. reaction buffer, 5 .mu.l 25 mM ATP/10 .mu.Ci
[.sup.33P]ATP (Amersham) and 5 .mu.l of 10.times. substrate.
[0309] 3. Start the reaction by the addition of 10 .mu.l of KDR (25
nM) in enzyme dilution buffer.
[0310] 4. Mix and incubate at room temperature for 15 minutes.
[0311] 5. Stop the reaction by the addition of 50 .mu.l of stop
solution.
[0312] 6. Incubate for 15 minutes at 4.degree. C.
[0313] 7. Transfer a 90 .mu.l aliquot to filter plate.
[0314] 8. Aspirate and wash 3 times with wash solution.
[0315] 9. Add 30 .mu.l of scintillation cocktail, seal plate and
count in a Wallace Microbeta scintillation counter.
Human Umbilical Vein Endothelial Cell Mitogenesis Assay
[0316] Expression of VEGF receptors that mediate mitogenic
responses to the growth factor is largely restricted to vascular
endothelial cells. Human umbilical vein endothelial cells (HUVECs)
in culture proliferate in response to VEGF treatment and can be
used as an assay system to quantify the effects of KDR kinase
inhibitors on VEGF stimulation. In the assay described, quiescent
HUVEC monolayers are treated with vehicle or test compound 2 hours
prior to addition of VEGF or basic fibroblast growth factor (BFGF).
The mitogenic response to VEGF or BFGF is determined by measuring
the incorporation of [.sup.3H]thymidine into cellular DNA.
Materials
[0317] HUVECs: HUVECs frozen as primary culture isolates are
obtained from Clonetics Corp. Cells are maintained in endothelial
growth medium (EGM; Clonetics) and are used for mitogenic assays at
passages 3-7.
[0318] Culture plates: NUNCLON 96-well polystyrene tissue culture
plates (NUNC #167008).
[0319] Assay medium: Dulbecco's modification of Eagle's medium
containing 1 g/ml glucose (low-glucose DMEM; Mediatech) plus 10%
(v/v) foetal bovine serum (Clonetics).
[0320] Test compounds: Working stock solutions of test compounds
are diluted serially in 100% dimethyl sulfoxide (DMSO) to 400 times
greater than their desired final concentrations. Final dilutions to
1.times. concentration are made directly into assay medium
immediately prior to addition to cells.
[0321] 10.times. growth factors: Solutions of human VEGF 165 (500
ng/ml; R&D Systems) and BFGF (10 ng/ml; R&D Systems) are
prepared in assay medium.
[0322] 10.times.[.sup.3H]thymidine: [Methyl-3H]thymidine (20
Ci/mmol; Dupont-NEN) is diluted to 80 .mu.Ci/ml in low-glucose
DMEM.
[0323] Cell wash medium: Hank's balanced salt solution (Mediatech)
containing 1 mg/ml bovine serum albumin (Boehringer-Mannheim).
[0324] Cell lysis solution: 1 N NaOH, 2% (w/v)
Na.sub.2CO.sub.3.
Method 1
[0325] HUVEC monolayers maintained in EGM are harvested by
trypsinisation and plated out at a density of 4000 cells per 100
.mu.l of assay medium per well in 96-well plates. Cells growth is
arrested for 24 hours at 37.degree. C. in a humidified atmosphere
containing 5% CO.sub.2.
Method 2
[0326] Growth-arrest medium is replaced by 100 .mu.l of assay
medium containing either vehicle (0.25% [v/v] DMSO) or the desired
final concentration of test compound. All determinations are
performed in triplicate. Cells are then incubated at 37.degree.
C./5% CO.sub.2 for 2 hours to allow test compounds to enter
cells.
Method 3
[0327] After the 2-hour pre-treatment period, cells are stimulated
by addition of 10 .mu.l/well of either assay medium, 10.times.VEGF
solution or 10.times.BFGF solution. Cells are then incubated at
37.degree. C./5% CO.sub.2.
Method 4
[0328] After 24 hours in the presence of growth factors,
10.times.[.sup.3H]thymidine (10 .mu.l/well) is added.
Method 5
[0329] Three days after addition of [.sup.3H]thymidine, medium is
removed by aspiration, and cells are washed twice with cell wash
medium (400 .mu.l/well followed by 200 .mu.l/well). The washed,
adherent cells are then solubilised by addition of cell lysis
solution (100 .mu.l/well) and warming to 37.degree. C. for 30
minutes. Cell lysates are transferred to 7 ml glass scintillation
vials containing 150 .mu.l of water. Scintillation cocktail (5
ml/vial) is added, and cell-associated radioactivity is determined
by liquid scintillation spectroscopy. According to these assays,
the compounds of the formula I are inhibitors of VEGF and are thus
suitable for the inhibition of angiogenesis, such as in the
treatment of ocular diseases, for example diabetic retinopathy, and
for the treatment of carcinomas, for example solid tumours. The
present compounds inhibit VEGF-stimulated mitogenesis of human
vascular endothelial cells in culture with IC 50 values of 0.01-5.0
.mu.M. These compounds also show selectivity over related tyrosine
kinases (for example, FGFR1 and the Src family; for relationship
between Src kinases and VEGFR kinases, see Eliceiri et al.,
Molecular Cell, Vol. 4, pp. 915-924, December 1999).
EXAMPLE 4
Injection Vials
[0330] A solution of 100 g of an active compound of the present
invention and 5 g of disodium hydrogenphosphate is adjusted to pH
6.5 in 3 l of double-distilled water using 2N hydrochloric acid,
sterile-filtered, dispensed into injection vials, lyophilized under
sterile conditions and aseptically sealed. Each injection vial
contains 5 mg of active compound.
EXAMPLE 5
Suppositories
[0331] A mixture of 20 g of an active compound of the present
invention is fused with 100 g of soya lecithin and 1400 g of cocoa
butter, poured into moulds and allowed to cool. Each suppository
contains 20 mg of active compound.
EXAMPLE 6
Solution
[0332] A solution of 1 g of an active compound of the present
invention, 9.38 g of NaH.sub.2PO.sub.4.2H.sub.2O, 28.48 g of
Na.sub.2HPO.sub.4.12H.sub.2O and 0.1 g of benzalkonium chloride in
940 ml of double-distilled water is prepared. It is adjusted to pH
6.8, made up to 1 L and sterilized by irradiation. This solution
can be used in the form of eye drops.
EXAMPLE 7
Ointment
[0333] 500 mg of an active compound of the present invention is
mixed with 99.5 g of petroleum jelly under aseptic conditions.
EXAMPLE 8
Tablets
[0334] A mixture of 1 kg of active compound of the present
invention, 4 kg of lactose, 1.2 kg of potato starch, 0.2 kg of talc
and 0.1 kg of magnesium stearate is compressed to give tablets in a
customary manner such that each tablet contains 10 mg of active
compound.
EXAMPLE 9
Coated Tablets
[0335] Analogously to Example E, tablets are pressed and are then
coated in a customary manner using a coating of sucrose, potato
starch, talc, tragacanth and colourant.
EXAMPLE 10
Capsules
[0336] 2 kg of active compound of the present invention are
dispensed into hard gelatin capsules in a customary manner such
that each capsule contains 20 mg of the active compound.
* * * * *