U.S. patent application number 09/791648 was filed with the patent office on 2002-03-14 for pharmaceutical agent containing rho kinase inhibitor.
Invention is credited to Kawahara, Toshio, Ono, Takashi, Satoh, Hiroyuki, Uehata, Masayoshi, Yamagami, Keiji.
Application Number | 20020032148 09/791648 |
Document ID | / |
Family ID | 16622116 |
Filed Date | 2002-03-14 |
United States Patent
Application |
20020032148 |
Kind Code |
A1 |
Uehata, Masayoshi ; et
al. |
March 14, 2002 |
Pharmaceutical agent containing Rho kinase inhibitor
Abstract
A Rho kinase inhibitor is provided as a novel pharmaceutical
agent, particularly as a therapeutic agent of hypertension, a
therapeutic agent of angina pectoris, a suppressive agent of
cerebrovascular contraction, a therapeutic agent of asthma, a
therapeutic agent of peripheral circulation disorder, a
prophylactic agent of immature birth, a therapeutic agent of
arteriosclerosis, an anti-cancer drug, an anti-inflammatory agent,
an immunosuppressant, a therapeutic agent of autoimmune disease, an
anti-AIDS drug, a contraceptive, a prophylactic agent of digestive
tract infection, a therapeutic agent of osteoporosis, a therapeutic
agent of retinopathy and a brain function improving drug. In
addition, the Rho kinase inhibitor is provided as a reagent and a
diagnostic.
Inventors: |
Uehata, Masayoshi;
(Iruma-shi, JP) ; Ono, Takashi; (Iruma-shi,
JP) ; Satoh, Hiroyuki; (Chikujo-gun, JP) ;
Yamagami, Keiji; (Iruma-shi, JP) ; Kawahara,
Toshio; (Chikujo-gun, JP) |
Correspondence
Address: |
WENDEROTH, LIND & PONACK, L.L.P.
2033 K STREET N. W.
SUITE 800
WASHINGTON
DC
20006-1021
US
|
Family ID: |
16622116 |
Appl. No.: |
09/791648 |
Filed: |
February 26, 2001 |
Related U.S. Patent Documents
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Application
Number |
Filing Date |
Patent Number |
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09791648 |
Feb 26, 2001 |
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09242261 |
Apr 19, 1999 |
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6218410 |
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09242261 |
Apr 19, 1999 |
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PCT/JP97/02793 |
Aug 8, 1997 |
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Current U.S.
Class: |
514/1 |
Current CPC
Class: |
A61K 31/551 20130101;
A61P 37/04 20180101; A61P 9/10 20180101; A61P 31/18 20180101; A61P
43/00 20180101; A61P 37/00 20180101; A61P 1/00 20180101; A61P 27/02
20180101; A61P 37/06 20180101; A61K 31/496 20130101; A61P 9/00
20180101; A61P 33/00 20180101; A61K 45/00 20130101; A61P 35/00
20180101; A61K 31/4545 20130101; A61K 31/4409 20130101; A61K 31/437
20130101; A61K 31/00 20130101 |
Class at
Publication: |
514/1 |
International
Class: |
A61K 031/00 |
Foreign Application Data
Date |
Code |
Application Number |
Aug 12, 1996 |
JP |
212409/1996 |
Claims
What is claimed is:
1. A pharmaceutical agent comprising a Rho kinase inhibitor.
2. A therapeutic agent of hypertension, comprising a Rho kinase
inhibitor.
3. A therapeutic agent of angina pectoris, comprising a Rho kinase
inhibitor.
4. A suppressive agent of cerebrovascular contraction, comprising a
Rho kinase inhibitor.
5. A therapeutic agent of asthma, comprising a Rho kinase
inhibitor.
6. A therapeutic agent of a peripheral circulation disorder,
comprising a Rho kinase inhibitor.
7. A therapeutic agent of arteriosclerosis, comprising a Rho kinase
inhibitor.
8. An anti-cancer drug comprising a Rho kinase inhibitor.
9. An anti-inflammatory agent comprising a Rho kinase
inhibitor.
10. An immunosuppressant comprising a Rho kinase inhibitor.
11. A therapeutic agent of an autoimmune disease, comprising a Rho
kinase inhibitor.
12. An anti-AIDS drug comprising a Rho kinase inhibitor.
13. A therapeutic agent of osteoporosis, comprising a Rho kinase
inhibitor.
14. A therapeutic agent of retinopathy, comprising a Rho kinase
inhibitor.
15. A brain function improving drug comprising a Rho kinase
inhibitor.
16. A prophylactic agent of immature birth, comprising a Rho kinase
inhibitor.
17. A contraceptive comprising a Rho kinase inhibitor.
18. A prophylactic agent of digestive tract infection, comprising a
Rho kinase inhibitor.
19. A pharmaceutical composition comprising a therapeutically
effective amount of a Rho kinase inhibitor and a pharmaceutically
acceptable additive.
20. A reagent comprising a Rho kinase inhibitor.
21. A diagnostic comprising a Rho kinase inhibitor.
22. A Rho kinase inhibitor comprising an amide compound of the
formula (I) 16wherein Ra is a group of the formula 17in the
formulas (a) and (b), R is hydrogen, alkyl or cycloalkyl,
cycloalkylalkyl, phenyl or aralkyl, which optionally have a
substituent on the ring, or a group of the formula 18wherein
R.sup.6 is hydrogen, alkyl or formula: --NR.sup.8NR.sup.9 wherein
R.sup.8 and R.sup.9 are the same or different and each is hydrogen,
alkyl, aralkyl or phenyl, R.sup.7 is hydrogen, alkyl, aralkyl,
phenyl, nitro or cyano, or R.sup.6 and R.sup.7 in combination show
a group forming a heterocycle optionally having, in the ring,
oxygen atom, sulfur atom or optionally substituted nitrogen atom,
R.sup.1 is hydrogen, alkyl or cycloalkyl, cycloalkylalkyl, phenyl
or aralky, which optionally have a substituent on the ring, or R
and R.sup.1 in combination form, together with the adjacent
nitrogen atom, a group forming a heterocycle optionally having, in
the ring, oxygen atom, sulfur atom or optionally substituted
nitrogen atom, R.sup.2 is hydrogen or alkyl, R.sup.3 and R.sup.4
are the same or different and each is hydrogen, alkyl, aralkyl,
halogen, nitro, amino, alkylamino, acylamino, hydroxy, alkoxy,
aralkyloxy, cyano, acyl, mercapto, alkylthio, aralkylthio, carboxy,
alkoxycarbonyl, carbamoyl, alkylcarbamoyl or azide, and A is a
group of the formula 19wherein R.sup.10 and R.sup.11 are the same
or different and each is hydrogen, alkyl, haloalkyl, aralkyl,
hydroxyalkyl, carboxy or alkoxycarbonyl, or R.sup.10 and R.sup.11
show a group which forms cycloalkyl in combination and l, m and n
are each 0 or an integer of 1-3, in the formula (c), L is hydrogen,
alkyl, aminoalkyl, mono or dialkylaminoalkyl, tetrahydrofurfuryl,
carbamoylalkyl, phthalimidoalkyl, amidino or a group of the formula
20wherein B is hydrogen, alkyl, alkoxy, aralkyl, aralkyloxy,
aminoalkyl, hydroxyalkyl, alkanoyloxyalkyl, alkoxycarbonylalkyl,
.alpha.-aminobenzyl, furyl, pyridyl, phenyl, phenylamino, styryl or
imidazopyridyl, Q.sup.1 is hydrogen, halogen, hydroxy, aralkyloxy
or thienylmethyl, W is alkylene, Q.sup.2 is hydrogen, halogen,
hydroxy or aralkyloxy, X is alkylene, Q.sup.3 is hydrogen, halogen,
hydroxy, alkoxy, nitro, amino, 2,3-dihydrofuryl or
5-methyl-3-oxo-2,3,4,5-tetrahydropyridazin-6-yl; and Y is a single
bond, alkylene or alkenylene, and in the formula (c), a broken line
is a single bond or a double bond, and R.sup.5 is hydrogen,
hydroxy, alkoxy, alkoxycarbonyloxy, alkanoyloxy or
aralyloxycarbonyloxy; Rb is a hydrogen, an alkyl, an aralkyl, an
aminoalkyl or a mono or dialkylaminoalkyl; and Rc is an optionally
substituted heterocycle containing nitrogen, an isomer thereof
and/or a pharmaceutically acceptable acid addition salt
thereof.
23. A therapeutic agent of hypertension caused by Rho kinase,
comprising a compound of the formula (I), an isomer thereof and/or
a pharmaceutically acceptable acid addition salt thereof.
24. A therapeutic agent of angina pectoris caused by Rho kinase,
comprising a compound of the formula (I), an isomer thereof and/or
a pharmaceutically acceptable acid addition salt thereof.
25. A suppressive agent of cerebrovascular contraction caused by
Rho kinase, comprising a compound of the formula (I), an isomer
thereof and/or a pharmaceutically acceptable acid addition salt
thereof.
26. A therapeutic agent of asthma caused by Rho kinase, comprising
a compound of the formula (I), an isomer thereof and/or a
pharmaceutically acceptable acid addition salt thereof.
27. A therapeutic agent of peripheral circulation disorder caused
by Rho kinase, comprising a compound of the formula (I), an isomer
thereof and/or a pharmaceutically acceptable acid addition salt
thereof.
28. A therapeutic agent of arteriosclerosis, comprising a compound
of the formula (I), an isomer thereof and/or a pharmaceutically
acceptable acid addition salt thereof.
29. An anti-cancer drug comprising a compound of the formula (I),
an isomer thereof and/or a pharmaceutically acceptable acid
addition salt thereof.
30. An anti-inflammatory agent comprising a compound of the formula
(I), an isomer thereof and/or a pharmaceutically acceptable acid
addition salt thereof.
31. An immunosuppressant comprising a compound of the formula (I),
an isomer thereof and/or a pharmaceutically acceptable acid
addition salt thereof.
32. A therapeutic agent of an autoimmune disease, comprising a
compound of the formula (I), an isomer thereof and/or a
pharmaceutically acceptable acid addition salt thereof.
33. An anti-AIDS drug comprising a compound of the formula (I), an
isomer thereof and/or a pharmaceutically acceptable acid addition
salt thereof.
34. A therapeutic agent of osteoporosis, comprising a compound of
the formula (I), an isomer thereof and/or a pharmaceutically
acceptable acid addition salt thereof.
35. A therapeutic agent of retinopathy, comprising a compound of
the formula (I), an isomer thereof and/or a pharmaceutically
acceptable acid addition salt thereof.
36. A brain function improving drug comprising a compound of the
formula (I), an isomer thereof and/or a pharmaceutically acceptable
acid addition salt thereof.
37. A prophylactic agent of immature birth, comprising a compound
of the formula (I), an isomer thereof and/or a pharmaceutically
acceptable acid addition salt thereof.
38. A contraceptive comprising a compound of the formula (I), an
isomer thereof and/or a pharmaceutically acceptable acid addition
salt thereof.
39. A prophylactic agent of digestive tract infection, comprising a
compound of the formula (I), an isomer thereof and/or a
pharmaceutically acceptable acid addition salt thereof.
40. A reagent having a Rho kinase inhibitory activity, comprising a
compound of the formula (I), an isomer thereof and/or a
pharmaceutically acceptable acid addition salt thereof.
41. A diagnostic of a disease caused by Rho kinase, comprising a
compound of the formula (I), an isomer thereof and/or a
pharmaceutically acceptable acid addition salt thereof.
42. A Rho kinase inhibitor containing a substituted
isoquinolinesulfonamide derivative of the formula (II) 21wherein
R.sup.12 is a hydrogen, a chlorine or a hydroxy, and when R.sup.12
is a hydrogen, Alk is an alkylene having 2 to 6 carbon atoms, which
optionally Has alkyl having 1 to 10 carbon atoms, aryl or aralkyl
as a substituent; R.sup.13 is a hydrogen; R.sup.14 is a hydrogen,
or a linear or branched alkyl having 1 to 6 carbon atoms, an aryl
or an aralkyl; R.sup.15 is a hydrogen, a linear or branched alkyl
having 1 to 6 carbon atoms, an aryl or an aralkyl, or a benzoyl, a
cinnamyl, a cinnamoyl, a furoyl or a group of the following formula
22wherein R.sup.16 is linear or branched alkyl having 1 to 6 carbon
atoms or a group of the following formula 23wherein R.sup.17 and
R.sup.18 are hydrogen or directly bonded to form alkylene having 2
to 4 carbon atoms; or R.sup.13 and R.sup.14 are directly bonded to
form alkylene having 4 or less carbon atoms, which is optionally
substituted by alkyl having 1 to 10 carbon atoms, phenyl or benzyl,
or R.sup.14 and R.sup.15 directly or in combination via oxygen atom
form a heterocycle together with the adjacent nitrogen atom, and
when R.sup.12 is a chlorine or a hydroxy, Alk is an alkylene having
2 to 6 carbon atoms, which is optionally substituted at the
hydrogen bonded to carbon by alkyl having 1 to 6 carbon atoms,
R.sup.13 and R.sup.14 are each a hydrogen, a linear or branched
alkyl having 1 to 6 carbon atoms or directly bonded to each other
to form ethylene or trimethylene, wherein hydrogen bonded to carbon
is optionally substituted by alkyl having 1 to 6 carbon atoms; or
R.sup.15 is a hydrogen, a linear or branched alkyl having 1 to 6
carbon atoms or an amidino, an isomer thereof and/or a
pharmaceutically acceptable acid addition salt thereof.
43. A therapeutic agent of hypertension caused by Rho kinase,
comprising a compound of the formula (II), an isomer thereof and/or
a pharmaceutically acceptable acid addition salt thereof.
44. A therapeutic agent of angina pectoris caused by Rho kinase,
comprising a compound of the formula (II), an isomer thereof and/or
a pharmaceutically acceptable acid addition salt thereof.
45. A suppressive agent of cerebrovascular contraction caused by
Rho kinase, comprising a compound of the formula (II), an isomer
thereof and/or a pharmaceutically acceptable acid addition salt
thereof.
46. A therapeutic agent of asthma caused by Rho kinase, comprising
a compound of the formula (II), an isomer thereof and/or a
pharmaceutically acceptable acid addition salt thereof.
47. A therapeutic agent of a peripheral circulation disorder,
comprising a compound of the formula (II), an isomer thereof and/or
a pharmaceutically acceptable acid addition salt thereof.
48. A therapeutic agent of arteriosclerosis, comprising a compound
of the formula (II), an isomer thereof and/or a pharmaceutically
acceptable acid addition salt thereof.
49. An anti-cancer drug comprising a compound of the formula (II),
an isomer thereof and/or a pharmaceutically acceptable acid
addition salt thereof.
50. A therapeutic agent of inflammation caused by Rho kinase,
comprising a compound of the formula (II), an isomer thereof and/or
a pharmaceutically acceptable acid addition salt thereof.
51. An immunosuppressant comprising a compound of the formula (II),
an isomer thereof and/or a pharmaceutically acceptable acid
addition salt thereof.
52. A therapeutic agent of an autoimmune disease, comprising a
compound of the formula (II), an isomer thereof and/or a
pharmaceutically acceptable acid addition salt thereof.
53. An anti-AIDS drug comprising a compound of the formula (II), an
isomer thereof and/or a pharmaceutically acceptable acid addition
salt thereof.
54. A therapeutic agent of osteoporosis, comprising a compound of
the formula (II), an isomer thereof and/or a pharmaceutically
acceptable acid addition salt thereof.
55. A therapeutic agent of retinopathy, comprising a compound of
the formula (II), an isomer thereof and/or a pharmaceutically
acceptable acid addition salt thereof.
56. A medicament for improving a brain function disorder caused by
Rho kinase, comprising a compound of the formula (II), an isomer
thereof and/or a pharmaceutically acceptable acid addition salt
thereof.
57. A prophylactic agent of immature birth, comprising a compound
of the formula (II), an isomer thereof and/or a pharmaceutically
acceptable acid addition salt thereof.
58. A contraceptive comprising a compound of the formula (II), an
isomer thereof and/or a pharmaceutically acceptable acid addition
salt thereof.
59. A prophylactic agent of digestive tract infection, comprising a
compound of the formula (II), an isomer thereof and/or a
pharmaceutically acceptable acid addition salt thereof.
60. A reagent having a Rho kinase inhibitory activity, comprising a
compound of the formula (II), an isomer thereof and/or a
pharmaceutically acceptable acid addition salt thereof.
61. A diagnostic of a disease caused by Rho kinase, comprising a
compound of the formula (II), an isomer thereof and/or a
pharmaceutically acceptable acid addition salt thereof.
62. A compound of the formula (III) 24wherein Rc' is an optionally
substituted heterocycle having nitrogen, which is other than
pyridine of Rc, and other symbols are as defined above, an isomer
thereof and/or a pharmaceutically acceptable acid addition salt
thereof.
63. The pharmaceutical agent of any of claims 1 to 18, comprising a
compound of the formula (III), an isomer thereof and/or a
pharmaceutically acceptable acid addition salt thereof as a Rho
kinase inhibitor.
64. The pharmaceutical composition of claim 19, comprising a
compound of the formula (III), an isomer thereof and/or a
pharmaceutically acceptable acid addition salt thereof as a Rho
kinase inhibitor.
65. The reagent of claim 20, comprising a compound of the formula
(III), an isomer thereof and/or a pharmaceutically acceptable acid
addition salt thereof as a Rho kinase inhibitor.
66. The diagnostic of claim 21, comprising a compound of the
formula (III), an isomer thereof and/or a pharmaceutically
acceptable acid addition salt thereof as a Rho kinase inhibitor.
Description
TECHNICAL FIELD
[0001] The present invention relates to treatment of various
diseases by the use of a Rho kinase inhibitor as a pharmaceutical
agent. Moreover, the present invention relates to use of a Rho
kinase inhibitor as a reagent or a diagnostic.
BACKGROUND ART
[0002] Ever since the discovery of Ras in 1981, a number of small
GTP binding proteins (small G proteins) similar to Ras have been
found, and many physiological functions they possess have been
studied. These small G proteins have a molecular weight of
20,000-30,000 and do not have a subunit structure. They all
specifically bind GDP and GTP, and hydrolyze the thus-bound GTP
(GTPase activity) (Hall, A., Science, 249, 635-640, 1990; Bourne,
H. R. et al., Nature, 349, 117-127, 1991).
[0003] To date, more than 50 kinds of genes encoding these small G
proteins have been found from yeast to mammals, forming a
superfamily. These small G proteins are largely divided into 5
groups of Ras, Rho, Rab, Arf and others, according to the
similarity of amino acid sequences.
[0004] Of these, Rho was named so because its gene isolated in the
form of cDNA from sea hare neuromuscle encodes a polypeptide having
about 35% homology with Ras (Ras homologue) (Madaule, P., Cell, 41,
31-40, 1985).
[0005] Rho is specifically ADP ribosylated by C3 enzyme, which is
one of the botulinum toxins, and Staphylococcal toxin EDIN, and
inactivated (Narumiya, S. and Morii, S., Cell Signal, 5, 9-19,
1993; Sekine, A. et al., J. Biol. Chem., 264, 8602-8605, 1989).
Hence, the C3 enzyme and EDIN were used to study the involvement of
Rho in cell functions from various aspects.
[0006] For example, phosphorylation by myosin light chain (MLC)
kinase is considered to enable actin.myosin interaction and
initiate contraction of smooth muscle, and the structure of smooth
muscle myosin phosphatase which dephosphorylates MLC has been
clarified (Shimizu, H. et al., J. Biol. Chem., 269, 30407-30411,
1994). It has been clarified that the activity of myosin
phosphatase is, like MLC kinase, under the control of the
intracellular signal transduction system and Rho is involved in
this mechanism. Moreover, an active Rho bound with GTP has been
found to enhance Ca-dependent contraction in a smooth muscle
skinned fiber specimen (Hirata, K., J. Biol. Chem., 267,8719-8722,
1992), thereby suggesting that the increase in Ca sensitivity in
smooth muscle contraction is caused by the inhibition of myosin
phosphatase activity via Rho.
[0007] In Swiss 3T3 cell and 3Y 1 cell, moreover, Rho-dependent
promotion of tyrosine phosphorylation (Kumagai, N. et al., J. Biol.
Chem., 270, 8466-8473, 1993) and activation of many kinds of
serine/threonine kinases (Kumagai, N. et al., FEBS Lett., 366,
11-16, 1995) have been acknowledged. From this, the presence of
plural protein kinases in the downstream of Rho in the signal
transduction pathway via Rho has been suggested and, actually,
ROC.alpha. (Leung, T. et al., J. Biol. Chem., 270, 29051-29054,
1995) [another name Rho-kinase, ROCK-II] and p160ROCK (Ishizaki, T.
et al., The EMBO J., 15(8), 1885-1893, 1996) [another name
ROC.beta., ROCK-I] have been reported as serine/threonine kinase
(Rho kinase) activated along with the activation of Rho. It has
been also reported that biological distribution of the both enzymes
is different (Nakagawa, O. et al., FEBS Lett. 392 189-193, 1996).
In addition, it has been reported that this Rho kinase directly
phosphorylates myosin phosphatase and inhibits its activity
(Kimura, K. et al., Science, 273, 245-248, 1996).
[0008] Rho has been documented to be responsible for the activation
of not only protein kinase but also lipid kinase (Zang, J. et al.,
J. Biol. Chem., 268, 22251-22254, 1993), and the presence of
phospholipase (PLD) activated by Rho has been also suggested
(Siddiqi, A. R. et al., J. Biol. Chem., 268, 24535-24538,
1995).
[0009] Control by Rho of the motility of Swiss 3T3 fibroblasts in
the presence of serum, motility of keratinocyte 303R by HGF and TPA
(12-O-tetradecanoylphorbol 13-acetate), spontaneously occurred and
chemoatractant mediated motility of neutrophils have been reported
(Takai, Y. et al., Trends Biochem. Sci., 20, 227-231, 1995), and
control of the permeation of liver cancer cell (MM1 cell), which is
one of the metastatic cancer models, through cultured mesothelial
layer by the activation of Rho has been reported (Yoshioka, K. et
al., FEBS Lett., 372, 25-28, 1995), thereby suggesting the
involvement of Rho in cell motility.
[0010] Meanwhile, in the cells derived from nerves, such as
neuroblastoma, PC-12 cells and the like, retraction of neurite and
rounding of the cell by lysophosphatidic acid, which is an
activation stimulant of Rho, have been acknowledged. Inasmuch as
this retraction can be inhibited by C3 enzyme treatment (Jalink, K.
et al., J. Cell Biol., 126, 801-810, 1994) and the formation of
ringed structure of podosome, which separates the site where
dissolution and absorption of bone take place in the clear zone of
osteoclast from the surrounding, is inhibited by C3 enzyme
treatment (Zhang, D. et al., J. Cell Sci., 108, 2285-2292, 1995), a
deep involvement of Rho in the morphological changes in cells has
been suggested.
[0011] In addition, C3 enzyme treatment reportedly inhibits
activation of an adhesion molecule such as LFA (leukocyte
function-associated antigen) and the like, and C3 enzyme treatment
reportedly inhibits proliferation of Swiss 3T3 fibroblasts
(Yamamoto, M. et al., Oncogene, 8, 1449-1455, 1993). Thus, Rho
reportedly controls cell adhesion and cell division via actin
cytoskeleton, and is also concerned with the transcription control
of c-fos gene (Hill, C. S. et al., Cell, 81, 1159-1170, 1995) and
transformation of cell (Khosravi-Far, R. et al., Mol. Cell Biol.,
15(11), 6443-6453, 1995).
[0012] In view of the inhibition of invasion of dysentery bacillus
into epithelial cells by C3 enzyme, a recent report has documented
the deep involvement of Rho in bacterial infection (Adam, T. et
al., The EMBO J., 15(13), 3315, 1996).
[0013] In pregnant rats, moreover, the levels of Rho and Rho kinase
are reported to be higher as compared to nonpregnant rats (Niiro,
N. et al., Biochem. Biophys. Res. Commun., 230, 356-359, 1997), and
deep involvement of Rho and Rho kinase in muscle contraction of
uterus for childbirth has been known. Further, integrin (Sueoka, K.
et al., Fertility & Sterility, 67(5) 799-811, 1997) considered
to be involved in the cell-cell and cell-extracellular matrix
adhesion during the stages of fertilization, embryogenesis and
embryonidation is known to be activated by Rho (Morii, N. et al.,
J. Biol. Chem., 267, 20921-20926, 1992).
[0014] Hence, it has been made clear that Rho is activated upon
receipt of signals from various cell membrane receptors and the
activated Rho functions as a molecule switch of a broad range of
cell phenomena, such as smooth muscle contraction, cell motility,
cell adhesion, morphological changes of cell, cell growth and the
like, via actomyosin system.
[0015] Smooth muscle contraction is significantly involved in the
disease states of hypertension, angina pectoris, cerebrovascular
contraction, asthma, peripheral circulation disorder, imminent
immature birth and the like; cell motility plays an important role
in invasion and metastasis of cancer, arteriosclerosis,
retinopathy, immune response and the like; cell adhesion is deeply
involved in metastasis of cancer, inflammation, autoimmune disease,
AIDS, fertilization and nidation of fertilized egg and the like;
morphological change of cell is deeply involved in brain function
disorder, osteoporosis, bacterial infection of digestive tract and
the like; and cell growth is deeply involved in cancer,
arteriosclerosis and the like. Therefore, a drug that blocks the
functions of Rho is considered to make a therapeutic agent for
these diseases in which Rho plays some role.
[0016] At present, however, only C3 enzyme and EDIN can inhibit the
actions of Rho. These are proteins which cannot permeate cytoplasm,
which prevents their development as a pharmaceutical agent.
[0017] On the other hand, inhibition of Rho kinase, which is
considered to be present downstream of the signal transduction
pathway via Rho, is considered to lead to the inhibition of
responses of various cell phenomena due to Rho. However, a specific
inhibitor of Rho kinase has not been known to date.
[0018] It is expected, therefore, that by searching a compound that
inhibits Rho kinase, such Rho kinase inhibitor will be an effective
agent for the prophylaxis and/or treatment of the above-mentioned
diseases and phenomena relating to Rho, such as hypertension,
angina pectoris, cerebrovascular contraction, asthma, peripheral
circulation disorder, immature birth, arteriosclerosis, cancer,
inflammation, immune disease, autoimmune disease, AIDS,
fertilization and nidation of fertilized egg, osteoporosis,
retinopathy, brain function disorder, bacterial infection of
digestive tract and the like.
[0019] The compound of the formula (I) is already known to be
useful as an agent for the prophylaxis and treatment of circulatory
disorder in coronary, cerebral, renal and peripheral arteries and
the like (e.g., a potent and long lasting therapeutic agent of
hypertension, angina pectoris, renal and peripheral circulation
disorder, and suppressive agent of cerebrovascular contraction and
the like), as well as a therapeutic agent of asthma (Japanese
Patent Unexamined Publication No. 62-89679, Japanese Patent
Unexamined Publication No. 3-218356, Japanese Patent Unexamined
Publication No. 4-273821, Japanese Patent Unexamined Publication
No. 5-194401, Japanese Patent Unexamined Publication No. 6-41080
and WO95/28387 and the like).
[0020] The compound of the formula (II) is already known to be
useful as a vasodilator, a therapeutic agent of hypertension, a
brain function improving agent, an anti-asthma agent, a heart
protection agent, a platelet aggregation inhibitor, a
psychosyndrome treating agent, an anti-inflammatory agent and an
agent for the prophylaxis and treatment of hyperviscosity syndrome
(Japanese Patent Unexamined Publication No. 57-200366, Japanese
Patent Unexamined Publication No. 61-227581, Japanese Patent
Unexamined Publication No. 2-256617, Japanese Patent Unexamined
Publication No. 4-264030, Japanese Patent Unexamined Publication
No. 6-56668, Japanese Patent Unexamined Publication No. 6-80569,
Japanese Patent Unexamined Publication No.6-293643, Japanese Patent
Unexamined Publication No. 7-41424 and Japanese Patent Unexamined
Publication No. 7-277979).
[0021] However, these compounds of the formula (I) or (II) are not
known to block the functions of Rho or to have Rho kinase
inhibitory action.
DISCLOSURE OF THE INVENTION
[0022] The present invention aims at providing a Rho kinase
inhibitor as a novel pharmaceutical agent. As a result of intensive
studies, the present inventors have found that a compound
inhibiting Rho kinase has an antihypertensive action, an
anti-angina pectoris action, a cerebrovascular contraction
suppressive action, an anti-asthma action, a peripheral circulation
improving action, an immature birth preventive action, an
anti-arteriosclerosis action, an anti-cancer action, an
antiinflammatory action, an immunosuppressive action, an autoimmune
disease improving action, an anti-AIDS action, a preventive action
on fertilization and nidation of fertilized egg, an osteoporosis
treating action, a retinopathy treating action, a brain function
improving action, a preventive action on bacterial infection of
digestive tract and that the Rho kinase inhibitor is useful as a
pharmaceutical agent, particularly as a therapeutic agent of
hypertension, a therapeutic agent of angina pectoris, a suppressive
agent of cerebrovascular contraction, a therapeutic agent of
asthma, a therapeutic agent of peripheral circulation disorder, a
prophylactic agent of immature birth, a therapeutic agent of
arteriosclerosis, an anti-cancer drug, an anti-inflammatory agent,
an immunosuppressant, a therapeutic agent of autoimmune disease, an
anti-AIDS drug, a therapeutic agent of osteoporosis, a therapeutic
agent of retinopathy, a brain function improving drug, a
contraceptive and a prophylactic agent of digestive tract
infection, which resulted in the completion of the present
invention.
[0023] It has been also found that a compound which inhibits Rho
kinase is useful as a reagent for the study of Rho and Rho kinase
and as a diagnostic of the diseases relating to those, which
resulted in the completion of the present invention.
[0024] Accordingly, the present invention provides the
following.
[0025] (1) A pharmaceutical agent containing a Rho kinase
inhibitor.
[0026] (2) A pharmaceutical agent containing a Rho kinase
inhibitor, which is at least one member selected from the group
consisting of a therapeutic agent of hypertension, a therapeutic
agent of angina pectoris, a suppressive agent of cerebrovascular
contraction, a therapeutic agent of asthma, a therapeutic agent of
peripheral circulation disorder, a therapeutic agent of
arteriosclerosis, an anti-cancer drug, an anti-inflammatory agent,
an immunosuppressant, a therapeutic agent of autoimmune disease, an
anti-AIDS drug, a therapeutic agent of osteoporosis, a therapeutic
agent of retinopathy, a brain function improving drug, a
prophylactic agent of immature birth, a contraceptive and a
prophylactic agent of digestive tract infection.
[0027] (3) A pharmaceutical composition containing a
therapeutically effective amount of a Rho kinase inhibitor and a
pharmaceutically acceptable additive.
[0028] (4) A reagent containing a Rho kinase inhibitor.
[0029] (5) A diagnostic containing a Rho kinase inhibitor.
[0030] (6) A Rho kinase inhibitor containing an amide compound of
the formula (I) 1
[0031] wherein
[0032] Ra is a group of the formula 2
[0033] in the formulas (a) and (b),
[0034] R is hydrogen, alkyl or cycloalkyl, cycloalkylalkyl, phenyl
or aralkyl, which optionally have a substituent on the ring, or a
group of the formula 3
[0035] wherein R.sup.6 is hydrogen, alkyl or formula:
--NR.sup.3NR.sup.9 wherein R.sup.8 and R.sup.9 are the same or
different and each is hydrogen, alkyl, aralkyl or phenyl, R.sup.7
is hydrogen, alkyl aralkyl, phenyl, nitro or cyano, or R.sup.6 and
R.sup.7 in combination show a group forming a heterocycle
optionally having, in the ring, oxygen atom, sulfur atom or
optionally substituted nitrogen atom,
[0036] R.sup.1 is hydrogen, alkyl or cycloalkyl, cycloalkylalkyl,
phenyl or aralkyl, which optionally have a substituent on the ring,
or
[0037] R and R.sup.1 in combination form, together with the
adjacent nitrogen atom, a group forming a heterocycle optionally
having, in the ring, oxygen atom, sulfur atom or optionally
substituted nitrogen atom,
[0038] R.sup.2 is hydrogen or alkyl,
[0039] R.sup.3 and R.sup.4 are the same or different and each is
hydrogen, alkyl, aralkyl, halogen, nitro, amino, alkylamino,
acylamino, hydroxy, alkoxy, aralkyloxy, cyano, acyl, mercapto,
alkylthio, aralkylthio, carboxy, alkoxycarbonyl, carbamoyl,
alkylcarbamoyl or azide, and
[0040] A is a group of the formula 4
[0041] wherein R.sup.10 and R.sup.11 are the same or different and
each is hydrogen, alkyl, haloalkyl, aralkyl, hydroxyalkyl, carboxy
or alkoxycarbonyl, or R.sup.10 and R.sup.11 show a group which
forms cycloalkyl in combination and l, m and n are each 0 or an
integer of 1-3,
[0042] in the formula (c),
[0043] L is hydrogen, alkyl, aminoalkyl, mono or dialkylaminoalkyl,
tetrahydrofurfuryl, carbamoylalkyl, phthalimidoalkyl, amidino or a
group of the formula 5
[0044] wherein B is hydrogen, alkyl, alkoxy, aralkyl, aralkyloxy,
aminoalkyl, hydroxyalkyl, alkanoyloxyalkyl, alkoxycarbonylalkyl,
.alpha.-aminobenzyl, furyl, pyridyl, phenyl, phenylamino, styryl or
imidazopyridyl,
[0045] Q.sup.1 is hydrogen, halogen, hydroxy, aralkyloxy or
thienylmethyl,
[0046] W is alkylene,
[0047] Q.sup.2 is hydrogen, halogen, hydroxy or aralkyloxy,
[0048] X is alkylene,
[0049] Q.sup.3 is hydrogen, halogen, hydroxy, alkoxy, nitro, amino,
2,3-dihydrofuryl or
5-methyl-3-oxo-2,3,4,5-tetrahydropyridazin-6-yl;
[0050] and Y is a single bond, alkylene or alkenylene, and
[0051] in the formula (c),
[0052] a broken line is a single bond or a double bond, and
[0053] R.sup.5 is hydrogen, hydroxy, alkoxy, alkoxycarbonyloxy,
alkanoyloxy or aralkyloxycarbonyloxy;
[0054] Rb is a hydrogen, an alkyl, an aralkyl, an aminoalkyl or a
mono- or dialkylaminoalkyl; and
[0055] Rc is an optionally substituted heterocycle containing
nitrogen, an isomer thereof and/or a pharmaceutically acceptable
acid addition salt thereof.
[0056] (7) A pharmaceutical agent containing a compound of the
formula (I), an isomer thereof and/or a pharmaceutically acceptable
acid addition salt thereof, which is a therapeutic agent of at
least one disease selected from the group consisting of
hypertension, angina pectoris, cerebrovascular contraction, asthma
and peripheral circulation disorder, which are caused by Rho
kinase.
[0057] (8) A pharmaceutical agent containing a compound of the
formula (I), an isomer thereof and/or a pharmaceutically acceptable
acid addition salt thereof, which is at least one therapeutic agent
selected from the group consisting of a therapeutic agent of
arteriosclerosis, an anti-cancer drug, an anti-inflammatory agent,
an immunosuppressant, a therapeutic agent of autoimmune disease, an
anti-AIDS drug, a therapeutic agent of osteoporosis, a therapeutic
agent of retinopathy, a brain function improving drug, a
prophylactic agent of immature birth, a contraceptive and a
prophylactic agent of digestive tract infection.
[0058] (9) A reagent having a Rho kinase inhibitory activity, which
contains a compound of the formula (I), an isomer thereof and/or a
pharmaceutically acceptable acid addition salt thereof.
[0059] (10) A diagnostic of a disease caused by Rho kinase, which
contains a compound of the formula (I), an isomer thereof and/or a
pharmaceutically acceptable acid addition salt thereof.
[0060] (11) A Rho kinase inhibitor containing a substituted
isoquinolinesulfonamide derivative of the formula (II) 6
[0061] wherein
[0062] R.sup.12 is a hydrogen, a chlorine or a hydroxy, and
[0063] when R.sup.12 is a hydrogen,
[0064] Alk is an alkylene having 2 to 6 carbon atoms, which
optionally has alkyl having 1 to 10 carbon atoms, aryl or aralkyl
as a substituent;
[0065] R.sup.13 is a hydrogen;
[0066] R.sup.14 is a hydrogen, or a linear or branched alkyl having
1 to 6 carbon atoms, an aryl or an aralkyl;
[0067] R.sup.15 is a hydrogen, a linear or branched alkyl having 1
to 6 carbon atoms, an aryl or an aralkyl, or a benzoyl, a cinnamyl,
a cinnamoyl, a furoyl or a group of the following formula 7
[0068] wherein R.sup.16 is linear or branched alkyl having 1 to 6
carbon atoms or a group of the following formula 8
[0069] wherein R.sup.17 and R.sup.18 are hydrogen or directly
bonded to form alkylene having 2 to 4 carbon atoms; or
[0070] R.sup.13 and R.sup.14 are directly bonded to form alkylene
having 4 or less carbon atoms, which is optionally substituted by
alkyl having 1 to 10 carbon atoms, phenyl or benzyl, or
[0071] R.sup.14 and R.sup.15 directly or in combination via oxygen
atom form a heterocycle together with the adjacent nitrogen atom,
and
[0072] when R.sup.12 is a chlorine or a hydroxy,
[0073] Alk is an alkylene having 2 to 6 carbon atoms, which is
optionally substituted at the hydrogen bonded to carbon by alkyl
having 1 to 6 carbon atoms,
[0074] R.sup.13 and R.sup.14 are each a hydrogen, a linear or
branched alkyl having 1 to 6 carbon atoms or directly bonded to
each other to form ethylene or trimethylene, wherein hydrogen
bonded to carbon is optionally substituted by alkyl having 1 to 6
carbon atoms; or
[0075] R.sup.15 is a hydrogen, a linear or branched alkyl having 1
to 6 carbon atoms or an amidino,
[0076] an isomer thereof and/or a pharmaceutically acceptable acid
addition salt thereof.
[0077] (12) A pharmaceutical agent containing a compound of the
formula (II), an isomer thereof and/or a pharmaceutically
acceptable acid addition salt thereof, which is a therapeutic agent
of at least one disease selected from the group consisting of
hypertension, angina pectoris, cerebrovascular contraction, asthma,
inflammation and, brain function disorder, which are caused by Rho
kinase.
[0078] (13) A pharmaceutical agent containing a compound of the
formula (II), an isomer thereof and/or a pharmaceutically
acceptable acid addition salt thereof, which is at least one
therapeutic agent selected from the group consisting of a
therapeutic agent of peripheral circulation disorder, a therapeutic
agent of arteriosclerosis, an anti-cancer drug, an
immunosuppressant, a therapeutic agent of autoimmune disease, an
anti-AIDS drug, a therapeutic agent of osteoporosis, a therapeutic
agent of retinopathy, a prophylactic agent of immature birth, a
contraceptive and a prophylactic agent of digestive tract
infection.
[0079] (14) A reagent having a Rho kinase inhibitory activity,
which contains a compound of the formula (II), an isomer thereof
and/or a pharmaceutically acceptable acid addition salt
thereof.
[0080] (15) A diagnostic for a disease caused by Rho kinase, which
contains a compound of the formula (II), an isomer thereof and/or a
pharmaceutically acceptable acid addition salt thereof.
[0081] (16) A compound of the formula (III) 9
[0082] wherein Rc' is an optionally substituted heterocycle having
nitrogen, which is other than pyridine of Rc, and other symbols are
as defined above, an isomer thereof and/or a pharmaceutically
acceptable acid addition salt thereof.
[0083] (17) The pharmaceutical agent of the above (1), containing a
compound of the formula (III), an isomer thereof and/or a
pharmaceutically acceptable acid addition salt thereof as a Rho
kinase inhibitor.
[0084] (18) A pharmaceutical agent containing a compound of the
formula (III), an isomer thereof and/or a pharmaceutically
acceptable acid addition salt thereof, which is at least one
therapeutic agent selected from the group consisting of a
therapeutic agent of hypertension, a therapeutic agent of angina
pectoris, a suppressive agent of cerebrovascular contraction, a
therapeutic agent of asthma, a therapeutic agent of peripheral
circulation disorder, a therapeutic agent of arteriosclerosis, an
anti-cancer drug, an anti-inflammatory agent, an immunosuppressant,
a therapeutic agent of autoimmune disease, an anti-AIDS drug, a
therapeutic agent of osteoporosis, a therapeutic agent of
retinopathy, a brain function improving drug, a prophylactic agent
of immature birth, a contraceptive and a prophylactic agent of
digestive tract infection.
[0085] (19) A pharmaceutical composition of the above (3),
containing a compound of the formula (III), an isomer thereof
and/or a pharmaceutically acceptable acid addition salt thereof as
a Rho kinase inhibitor.
[0086] (20) A reagent having a Rho kinase inhibitory activity,
which contains a compound of the formula (III), an isomer thereof
and/or a pharmaceutically acceptable acid addition salt thereof as
a Rho kinase inhibitor.
[0087] (21) A diagnostic for a disease caused by Rho kinase, which
contains a compound of the formula (III), an isomer thereof and/or
a pharmaceutically acceptable acid addition salt thereof.
[0088] (22) A method for treating a disease based on inhibition of
Rho kinase, comprising administering a pharmaceutically effective
amount of a Rho kinase inhibitor to a patient.
[0089] (23) The treating method of the above (22), wherein the
disease treatable by the inhibition of the Rho kinase is at least
one disease selected from the group consisting of hypertension,
angina pectoris, cerebrovascular contraction, asthma, a peripheral
circulation disorder, arteriosclerosis, cancer, an inflammation, an
immune disease, an autoimmune disease, AIDS, osteoporosis,
retinopathy, a brain function disorder, immature birth,
fertilization and nidation of fertilized egg and infection of
digestive tract.
[0090] (24) A method for treating at least one disease selected
from the group consisting of hypertension, angina pectoris,
cerebrovascular contraction, asthma and a peripheral circulation
disorder, which are caused by Rho kinase, and arteriosclerosis,
cancer, inflammation, immune disease, autoimmune disease, AIDS,
osteoporosis, retinopathy, brain function disorder, immature birth,
fertilization and nidation of fertilized egg and infection of
digestive tract, which comprises administering a pharmaceutically
effective amount of a compound of the formula (I), an isomer
thereof and/or a pharmaceutically acceptable acid addition salt
thereof.
[0091] (25) A method for treating at least one disease selected
from the group consisting of hypertension, angina pectoris,
cerebrovascular contraction, asthma, inflammation and brain
function disorder, which are caused by Rho kinase, and a peripheral
circulation disorder, arteriosclerosis, cancer, immune disease,
autoimmune disease, AIDS, osteoporosis, retinopathy, immature
birth, fertilization and nidation of fertilized egg and infection
of digestive tract, which comprises administering a
pharmaceutically effective amount of a compound of the formula
(II), an isomer thereof and/or a pharmaceutically acceptable acid
addition salt thereof.
[0092] (26) A method for treating at least one disease selected
from the group consisting of hypertension, angina pectoris,
cerebrovascular contraction, asthma, peripheral circulation
disorder, arteriosclerosis, cancer, inflammation, immune disease,
autoimmune disease, AIDS, osteoporosis, retinopathy, brain function
disorder, immature birth, fertilization and nidation of fertilized
egg and infection of digestive tract, which comprises administering
a pharmaceutically effective amount of a compound of the formula
(III), an isomer thereof and/or a pharmaceutically acceptable acid
addition salt thereof.
[0093] (27) Use of a Rho kinase inhibitor for the production of a
therapeutic agent of a disease treatable by inhibiting Rho
kinase.
[0094] (28) The use of a Rho kinase inhibitor of the above (27),
wherein the disease treatable by the inhibition of Rho kinase is at
least one member selected from the group consisting of
hypertension, angina pectoris, cerebrovascular contraction, asthma,
peripheral circulation disorder, arteriosclerosis, cancer,
inflammation, immune disease, autoimmune disease, AIDS,
osteoporosis, retinopathy, brain function disorder, immature birth,
fertilization and nidation of fertilized egg and infection of
digestive tract.
[0095] (29) The use of a compound of the formula (I), an isomer
thereof and/or a pharmaceutically acceptable acid addition salt
thereof for the production of a therapeutic agent of at least one
disease selected from the group consisting of hypertension, angina
pectoris, cerebrovascular contraction, asthma and peripheral
circulation disorder caused by Rho kinase, and arteriosclerosis,
cancer, inflammation, immune disease, autoimmune disease, AIDS,
osteoporosis, retinopathy, brain function disorder, immature birth,
fertilization and nidation of fertilized egg and infection of
digestive tract.
[0096] (30) Use of a compound of the formula (II), an isomer
thereof and/or a pharmaceutically acceptable acid addition salt
thereof for the production of a therapeutic agent of at least one
disease selected from the group consisting of hypertension, angina
pectoris, cerebrovascular contraction, asthma, inflammation and
brain function disorder caused by Rho kinase, and peripheral
circulation disorder, arteriosclerosis, cancer, immune disease,
autoimmune disease, AIDS, osteoporosis, retinopathy, immature
birth, fertilization and nidation of fertilized egg and infection
of digestive tract.
[0097] (31) Use of a compound of the formula (III), an isomer
thereof and/or a pharmaceutically acceptable acid addition salt
thereof for the production of a therapeutic agent of at least one
disease selected from the group consisting of hypertension, angina
pectoris, cerebrovascular contraction, asthma, peripheral
circulation disorder, arteriosclerosis, cancer, inflammation,
immune disease, autoimmune disease, AIDS, osteoporosis,
retinopathy, brain function disorder, immature birth, fertilization
and nidation of fertilized egg and infection of digestive
tract.
[0098] (32) A commercial package comprising a Rho kinase inhibitor
and a written matter associated therewith, the written matter
stating that the Rho kinase inhibitor can or should be used for
treating at least one disease selected from the group consisting of
hypertension, angina pectoris, cerebrovascular contraction, asthma,
peripheral circulation disorder, arteriosclerosis, cancer,
inflammation, immune disease, autoimmune disease, AIDS,
osteoporosis, retinopathy, brain function disorder, immature birth,
fertilization and nidation of fertilized egg and infection of
digestive tract.
[0099] (33) A commercial package comprising a compound of the
formula (I), an isomer thereof and/or a pharmaceutically acceptable
acid addition salt thereof and a written matter associated
therewith, the written matter stating that the compound can or
should be used for treating at least one disease selected from the
group consisting of hypertension, angina pectoris, cerebrovascular
contraction, asthma and peripheral circulation disorder, which are
caused by Rho kinase, and arteriosclerosis, cancer, inflammation,
immune disease, autoimmune disease, AIDS, osteoporosis,
retinopathy, brain function disorder, immature birth, fertilization
and nidation of fertilized egg and infection of digestive
tract.
[0100] (34) A commercial package comprising a compound of the
formula (II), an isomer thereof and/or a pharmaceutically
acceptable acid addition salt thereof and a written matter
associated therewith, the written matter stating that the compound
can or should be used for treating at least one disease selected
from the group consisting of hypertension, angina pectoris,
cerebrovascular contraction, asthma, inflammation and brain
function disorder, which are caused by Rho kinase, and peripheral
circulation disorder, arteriosclerosis, cancer, immune disease,
autoimmune disease, AIDS, osteoporosis, retinopathy, immature
birth, fertilization and nidation of fertilized egg and infection
of digestive tract.
[0101] (35) A commercial package comprising a compound of the
formula (III), an isomer thereof and/or a pharmaceutically
acceptable acid addition salt thereof and a written matter
associated therewith, the written matter stating that the compound
can or should be used for treating at least one disease selected
from the group consisting of hypertension, angina pectoris,
cerebrovascular contraction, asthma, peripheral circulation
disorder, arteriosclerosis, cancer, inflammation, immune disease,
autoimmune disease, AIDS, osteoporosis, retinopathy, brain function
disorder, immature birth, fertilization and nidation of fertilized
egg and infection of digestive tract.
DETAILED DESCRIPTION OF THE INVENTION
[0102] The Rho kinase inhibitory action, antihypertensive action,
anti-angina pectoris action, cerebrovascular contraction
suppressive action, anti-asthma action, peripheral circulation
improving action, immature birth preventive action,
anti-arteriosclerosis action, anti-cancer action, antiinflammatory
action, immunosuppressive action, autoimmune disease improving
action, anti-AIDS action, preventive action of fertilization and
nidation of fertilized egg, preventive action on bacterial
infection of digestive tract, osteoporosis treating action,
retinopathy treating action and brain function improving action of
the present invention can be confirmed by Rho kinase inhibitory
activity, vasohypotonic action, trachea relaxing action, peripheral
blood flow increasing action, cell adhesion induction inhibitory
action, malignant tumor metastasis inhibitory action, bone
resorption inhibitory action, mouse allogenic MLR inhibitory
activity, tumor cell proliferation inhibitory action, angiogenesis
inhibitory action, vascular smooth muscle cell proliferation
inhibitory action and the like.
[0103] The disease relating to Rho, on which the inventive Rho
kinase inhibitor is effective include, for example, disease
symptoms of hypertension, angina pectoris, cerebrovascular
contraction, asthma, peripheral circulation disorder, immature
birth, arteriosclerosis, cancer, inflammation, immune disease,
autoimmune disease, AIDS, bacterial infection of digestive tract,
osteoporosis, retinopathy, brain function disorder and the like, as
well as biological phenomena such as fertilization and nidation of
fertilized egg.
[0104] As used herein, by the Rho kinase of the present invention
is meant serine/threonine kinase activated along with the
activation of Rho, which is exemplified by the aforementioned
ROC.alpha.(ROCKII), p160ROCK(ROC.beta., ROCK-I) and other proteins
having serine/threonine kinase activity.
[0105] Cancer includes bone marrow leukemia, lymphocytic leukemia,
gastric cancer, colon cancer, lung cancer, pancreatic cancer, liver
cancer, cancer of esophangus, ovarian cancer, breast cancer, skin
cancer, cervical cancer, orchioncus, neuroblastoma, urinary
epithelial cancer, multiple myeloma, uterine cancer, melanoma,
cerebral tumor and the like, and anti-cancer means inhibition of
formation, infiltration, metastasis, growth and the like of these
tumors.
[0106] The immune disease includes allergic diseases, rejection in
organ transplantation and the like.
[0107] The autoimmune disease includes articular rheumatism,
systemic lupus erythematodes, Sjogren's disease, multiple
sclerosis, myasthenia gravis, type I diabetes, endocrine
ophthalmopathy, primary biliary cirrhosis, Crohn's disease,
glomerulonephritis, sarcoidosis, psoriasis, pemphigus, hyoplastic
anemia, essential thrombocytopenic purpura and the like.
[0108] Bacterial infection of digestive tract means various
diseases caused by the invasion of Salmonella, dysentery bacillus,
intestinal pathogenic Escherichia coli and the like into intestinal
mucosa epithelial cells.
[0109] Retinopathy means angiopathic retinopathy, arteriosclerosis
retinopathy, central angiospastic retinopathy, central serous
retinopathy, circinate retinopathy, diabetic retinopathy,
dysproteinemic retinopathy, hypertensive retinopathy, leukemic
retinopathy, lipemic retinopathy, proliferative retinopathy, renal
retinopathy, sickle retinopathy, toxemic retinopathy of pregnancy
and the like.
[0110] Brain function disorder includes psychotic condition due to
cerebral hemorrhage, cerebral thrombus, cerebral embolus,
subarachnoid hemorrhage, transient cerebral ischemic stroke,
hypertensive encephalopathy, cerebral arteriosclerosis, subdural
hematoma, extradural hematoma, cerebral hypoxia, cerebral edema,
cerebritis, cerebral tumor, external injury in head, mental
disease, metabolite poisoning, drug poisoning, temporal respiratory
arrest, deep anesthesia during operation, physical disorder and the
like, and sequelae, decreased attention, hyperactivity, logopathy,
delayed mental development, lethe, dementia (inclusive of
wandering, nocturnal delirium, aggressive behavior and the like
associated with dementia) caused by the above-mentioned
diseases.
[0111] Therefore, the Rho kinase inhibitor of the present invention
is effective as a pharmaceutical agent, particularly as an agent
for the prophylaxis and treatment of these diseases caused by Rho,
such as a therapeutic agent of hypertension, a therapeutic agent of
angina pectoris, a suppressive agent of cerebrovascular
contraction, a therapeutic agent of asthma, a therapeutic agent of
peripheral circulation disorder, a prophylactic agent of immature
birth, a therapeutic agent of arteriosclerosis, an anti-cancer
drug, an anti-inflammatory agent, an immunosuppressant, a
therapeutic agent of autoimmune disease, an anti-AIDS drug, a
contraceptive, a prophylactic agent of digestive tract infection, a
therapeutic agent of osteoporosis, a therapeutic agent of
retinopathy and a brain function improving drug.
[0112] The compounds of the formula (I) and the formula (II) have
high affinity for Rho kinase. Thus, the radioactive substance
(radio ligand) thereof are industrially useful as a selective radio
ligand of Rho kinase. The compounds of the formula (I) and the
formula (II) and modified compounds thereof (e.g., radio ligand of
these compounds and the like), which are Rho kinase inhibitors, are
useful as reagents for the study of Rho and Rho kinase and as
diagnostics of the diseases relating to them.
[0113] The compound to be used as the Rho kinase inhibitor of the
present invention may be any as long as it has a Rho kinase
inhibitory action. For example, the compounds of the formula (I)
and the formula (II) are used.
[0114] In the present specification, each symbol of the formula (I)
is defined as follows.
[0115] Alkyl at R and R.sup.1 is linear or branched alkyl having 1
to 10 carbon atoms, which is exemplified by methyl, ethyl, propyl,
isopropyl, butyl, isobutyl, sec-butyl, tert-butyl, pentyl, hexyl,
heptyl, octyl, nonyl, decyl and the like, with preference given to
alkyl having 1 to 4 carbon atoms.
[0116] Cycloalkyl at R and R.sup.1 has 3 to 7 carbon atoms and is
exemplified by cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl,
cycloheptyl and the like.
[0117] Cycloalkylalkyl at R and R.sup.1 is that wherein the
cycloalkyl moiety is the above-mentioned cycloalkyl having 3 to 7
carbon atoms and the alkyl moiety is linear or branched alkyl
having 1 to 6 carbon atoms (e.g., methyl, ethyl, propyl, isopropyl,
butyl, pentyl, hexyl and the like), which is exemplified by
cyclopropylmethyl, cyclobutylmethyl, cyclopentylmethyl,
cyclohexylmethyl, cycloheptylmethyl, cyclopropylethyl,
cyclopentylethyl, cyclohexylethyl, cycloheptylethyl,
cyclopropylpropyl, cyclopentylpropyl, cyclohexylpropyl,
cycloheptylpropyl, cyclopropylbutyl, cyclopentylbutyl,
cyclohexylbutyl, cycloheptylbutyl, cyclopropylhexyl,
cyclopentylhexyl, cyclohexylhexyl, cycloheptylhexyl and the
like.
[0118] Aralkyl at R and R.sup.1 is that wherein alkyl moiety is
alkyl having 1 to 4 carbon atoms and is exemplified by phenylalkyl
such as benzyl, 1-phenylethyl, 2-phenylethyl, 3-phenylpropyl,
4-phenylbutyl and the like.
[0119] The substituent of optionally substituted cycloalkyl,
cycloalkylalkyl, phenyl and aralkyl on the ring at R and R.sup.1 is
halogen (e.g., chlorine, bromine, fluorine and iodine), alkyl (same
as alkyl at R and R.sup.1), alkoxy (linear or branched alkoxy
having 1 to 6 carbon atoms, such as methoxy, ethoxy, propoxy,
isopropoxy, butoxy, isobutoxy, sec-butoxy, tert-butoxy, pentyloxy,
hexyloxy and the like), aralkyl (same as aralkyl at R and R.sup.1)
or haloalkyl (alkyl at R and R.sup.1 which is substituted by 1-5
halogen, and exemplified by fluoromethyl, difluoromethyl,
trifluoromethyl, 2,2,2-trifluoroethyl, 2,2,3,3,3-pentafluoropropyl
and the like), nitro, amino, cyano, azide and the like.
[0120] The group formed by R and R' in combination together with
the adjacent nitrogen atom, which forms a heterocycle optionally
having, in the ring, oxygen atom, sulfur atom or optionally
substituted nitrogen atom is preferably a 5 or 6-membered ring and
bonded ring thereof. Examples thereof include 1-pyrrolidinyl,
piperidino, 1-piperazinyl, morpholino, thiomorpholino,
1-imidazolyl, 2,3-dihydrothiazol-3-yl and the like. The substituent
of the optionally substituted nitrogen atom is exemplified by
alkyl, aralkyl, haloalkyl and the like. As used herein, alkyl,
aralkyl and haloalkyl are as defined for R and R.sup.1.
[0121] Alkyl at R.sup.2 is as defined for R and R.sup.1.
[0122] Halogen, alkyl, alkoxy and aralkyl at R.sup.3 and R.sup.4
are as defined for R and R.sup.1.
[0123] Acyl at R.sup.3 and R.sup.4 is alkanoyl having 2 to 6 carbon
atoms (e.g., acetyl, propionyl, butyryl, valeryl, pivaloyl and the
like), benzoyl or phenylalkanoyl wherein the alkanoyl moiety has 2
to 4 carbon atoms (e.g., phenylacetyl, phenylpropionyl,
phenylbutyryl and the like).
[0124] Alkylamino at R.sup.3 and R.sup.4 is that wherein the alkyl
moiety is alkylamino having linear or branched alkyl having 1 to 6
carbon atoms. Examples thereof include methylamino, ethylamino,
propylamino, isopropylamino, butylamino, isobutylamino,
sec-butylamino, tert-butylamino, pentylamino, hexylamino and the
like.
[0125] Acylamino at R.sup.3 and R.sup.4 is that wherein acyl moiety
is alkanoyl having 2 to 6 carbon atoms, benzyl or the alkanoyl
moiety is phenylalkanoyl having 2 to 4 carbon atoms and the like,
which is exemplified by acetylamino, propionylamino, butyrylamino,
valerylamino, pivaloylamino, benzoylamino, phenylacetylamino,
phenylpropionylamino, phenylbutyrylamino and the like.
[0126] Alkylthio at R.sup.3 and R.sup.4 is that wherein the alkyl
moiety is linear or branched alkyl having 1 to 6 carbon atoms,
which is exemplified by methylthio, ethylthio, propylthio,
isopropylthio, butylthio, isobutylthio, sec-butylthio,
tert-butylthio, pentylthio, hexylthio and the like.
[0127] Aralkyloxy at R.sup.3 and R.sup.4 is that wherein the alkyl
moiety is alkyl having 1 to 4 carbon atoms, which is exemplified by
benzyloxy, 1-phenylethyloxy, 2-phenylethyloxy, 3-phenylpropyloxy,
4-phenylbutyloxy and the like.
[0128] Aralkylthio at R.sup.3 and R.sup.4 is that wherein the alkyl
moiety is alkyl having 1 to 4 carbon atoms, which is exemplified by
benzylthio,
1-phenylethylthio,2-phenylethylthio,3-phenylpropylthio,4-phenylbutylthio
and the like.
[0129] Alkoxycarbonyl at R.sup.3 and R.sup.4 is that wherein the
alkoxy moiety is linear or branched alkoxy having 1 to 6 carbon
atoms, which is exemplified by methoxycarbonyl, ethoxycarbonyl,
propoxycarbonyl, isopropoxycarbonyl, butoxycarbonyl,
isobutoxycarbonyl, sec-butoxycarbonyl, tert-butoxycarbonyl,
pentyloxycarbonyl, hexyloxycarbonyl and the like.
[0130] Alkylcarbamoyl at R.sup.3 and R.sup.4 is carbamoyl mono- or
di-substituted by alkyl having 1 to 4 carbon atoms, which is
exemplified by methylcarbamoyl, dimethylcarbamoyl, ethylcarbamoyl,
diethylcarbamoyl, propylcarbamoyl, dipropylcarbamoyl,
butylcarbamoyl, dibutylcarbamoyl and the like.
[0131] Alkoxy at R.sup.5 is as defined for R and R.sup.1.
[0132] Alkoxycarbonyloxy at R.sup.5 is that wherein the alkoxy
moiety is linear or branched alkoxy having 1 to 6 carbon atoms,
which is exemplified by methoxycarbonyloxy, ethoxycarbonyloxy,
propoxycarbonyloxy, isopropoxycarbonyloxy, butoxycarbonyloxy,
isobutoxycarbonyloxy, sec-butoxycarbonyloxy,
tert-butoxycarbonyloxy, pentyloxycarbonyloxy, hexyloxycarbonyloxy
and the like.
[0133] Alkanoyloxy at R.sup.5 is that wherein the alkanoyl moiety
is alkanoyl having 2 to 6 carbon atoms, which is exemplified by
acetyloxy, propionyloxy, butyryloxy, valeryloxy, pivaloyloxy and
the like.
[0134] Aralkyloxycarbonyloxy at R.sup.5 is that wherein the aralkyl
moiety is aralkyl having C.sub.1-C.sub.4 alkyl, which is
exemplified by benzyloxycarbonyloxy, 1-phenylethyloxycarbonyloxy,
2-phenylethyloxycarbonyloxy, 3-phenylpropyloxycarbonyloxy,
4-phenylbutyloxycarbonyloxy and the like.
[0135] Alkyl at R.sup.6 is as defined for R and R.sup.1; alkyl at
R.sup.8 and R.sup.9 is as defined for R and R.sup.1; and aralkyl at
R.sup.8 and R.sup.9 is as defined for R and R.sup.1.
[0136] Alkyl at R.sup.7 is as defined for R and R.sup.1 and aralkyl
at R.sup.7 is as defined for R and R.sup.1.
[0137] The group formed by R.sup.6 and R.sup.7 in combination,
which forms a heterocycle optionally having, in the ring, oxygen
atom, sulfur atom or optionally substituted nitrogen atom, is
imidazol-2-yl, thiazol-2-yl, oxazol-2-yl, imidazolin-2-yl,
3,4,5,6-tetrahydropyridin-2-yl, 3,4,5,6-tetrahydropyrimidin-2-yl,
1,3-oxazolin-2-yl, 1,3-thiazolin-2-yl or optionally substituted
benzoimidazol-2-yl, benzothiazol-2-yl, benzoxazol-2-yl and the like
having a substituent such as halogen, alkyl, alkoxy, haloalkyl,
nitro, amino, phenyl, aralkyl and the like. As used herein,
halogen, alkyl, alkoxy, haloalkyl and aralkyl are as defined for R
and R.sup.1.
[0138] The substituent of the above-mentioned optionally
substituted nitrogen atom is exemplified by alkyl, aralkyl,
haloalkyl and the like. As used herein, alkyl, aralkyl and
haloalkyl are as defined for R and R.sup.1.
[0139] Hydroxyalkyl at R.sup.10 and R.sup.11 is linear or branched
alkyl having 1 to 6 carbon atoms which is substituted by 1 to 3
hydroxy, which is exemplified by hydroxymethyl, 2-hydroxyethyl,
1-hydroxyethyl, 3-hydroxypropyl, 4-hydroxybutyl and the like. Alkyl
at R.sup.10 and R.sup.11 is as defined for R and R.sup.1; haloalkyl
and alkoxycarbonyl at R.sup.10 and R.sup.11 are as defined for R
and R.sup.1; aralkyl at R.sup.10 and R.sup.11 is as defined for R
and R.sup.1; and cycloalkyl formed by R.sup.10 and R.sup.11 in
combination is the same as cycloalkyl at R and R.sup.1.
[0140] Alkyl at L is as defined for R and R.sup.1.
[0141] Aminoalky at L is a linear or branched alkyl having 1 to 6
carbon atoms, which is substituted by amino, which is exemplified
by aminomethyl, 2-aminoethyl, 1-aminoethyl, 3-aminopropyl,
4-aminobutyl, 5-aminopentyl, 6-aminohexyl and the like.
[0142] Mono- or dialkylaminoalkyl at L is mono- or di-substituted
aminoalkyl with alkyl having 1 to 4 carbon atoms, which is
exemplified by methylaminomethyl, dimethylaminomethyl,
ethylaminomethyl, diethylaminomethyl, propylaminomethyl,
dipropylaminomethyl, butylaminomethyl, dibutylaminomethyl,
2-dimethylaminoethyl, 2-diethylaminoethyl and the like.
[0143] Carbamoylalkyl at L is linear or branched alkyl having 1 to
6 carbon atoms substituted by carbamoyl, which is exemplified by
carbamoylmethyl, 2-carbamoylethyl, 1-carbamoylethyl,
3-carbamoylpropyl, 4-carbamoylbutyl, 5-carbamoylpentyl,
6-carbamoylhexyl and the like.
[0144] Phthalimidoalkyl at L is linear or branched alkyl having 1
to 6 carbon atoms, which is substituted by phthalimide. Examples
thereof include phthalimidomethyl, 2-phthalimidoethyl,
1-phthalimidoethyl, 3-phthalimidopropyl, 4-phthalimidobutyl,
5-phthalimidopentyl, 6-phthalimidohexyl and the like.
[0145] Alkyl at B is as defined for R and R.sup.1.
[0146] Alkoxy at B is as defined for R and R.sup.1.
[0147] Aralkyl at B is as defined for R and R.sup.1.
[0148] Aralkyloxy at B is as defined for R.sup.3 and R.sup.4.
[0149] Aminoalkyl at B is as defined for L.
[0150] Hydroxyalkyl at B is as defined for R.sup.10 and
R.sup.11.
[0151] Alkanoyloxyalkyl at B is that wherein linear or branched
alkyl having 1 to 6 carbon atoms is substituted by alkanoyloxy
having alkanoyl moiety having 2 to 6 carbon atoms, which is
exemplified by acetyloxymethyl, propionyloxymethyl,
butyryloxymethyl, valeryloxymethyl, pivaloyloxymethyl,
acetyloxyethyl, propionyloxyethyl, butyryloxyethyl,
valeryloxyethyl, pivaloyloxyethyl and the like.
[0152] Alkoxycarbonylalkyl at B is that wherein linear or branched
alkyl having 1 to 6 carbon atoms is substituted by alkoxycarbonyl
having alkoxy moiety having 1 to 6 carbon atoms, which is
exemplified by methoxycarbonylmethyl, ethoxycarbonylmethyl,
propoxycarbonylmethyl, isopropoxycarbonylmethyl,
butoxycarbonylmethyl, isobutoxycarbonylmethyl,
sec-butoxycarbonylmethyl, tert-butoxycarbonylmethyl,
pentyloxycarbonylmethyl, hexyloxycarbonylmethyl,
methoxycarbonylethyl, ethoxycarbonylethyl, propoxycarbonylethyl,
isopropoxycarbonylethyl, butoxycarbonylethyl,
isobutoxycarbonylethyl, sec-butoxycarbonylethyl,
tert-butoxycarbonylethyl, pentyloxycarbonylethyl,
hexyloxycarbonylethyl and the like.
[0153] Halogen at Q.sup.1, Q.sup.2 and Q.sup.3 is as defined for R
and R.sup.1.
[0154] Aralkyloxy at Q.sup.1 and Q.sup.2 is as defined for R.sup.3
and R.sup.4.
[0155] Alkoxy at Q.sup.3 is as defined for R and R.sup.1.
[0156] Alkylene at W, X and Y is linear or branched alkylene having
1 to 6 carbon atoms, which is exemplified by methylene, ethylene,
trimethylene, propylene, tetramethylene, pentamethylene,
hexamethylene and the like.
[0157] Alkenylene at Y is linear or branched alkenylene having 2 to
6 carbon atoms, which is exemplified by vinylene, propenylene,
butenylene, pentenylene and the like.
[0158] Alkyl at Rb is as defined for R and R.sup.1.
[0159] Aralkyl at Rb is as defined for R and R.sup.1.
[0160] Aminoalkyl at Rb is as defined for L.
[0161] Mono- or dialkylaminoalkyl at Rb is as defined for L.
[0162] The heterocycle when single ring containing nitrogen at Rc
is pyridine, pyrimidine, pyridazine, triazine, pyrazole, triazole
and the like, and when it is a condensed ring, it is exemplified by
pyrrolopyridine (e.g., 1H-pyrrolo[2,3-b]pyridine,
1H-pyrrolo[3,2-b]pyridi- ne, 1H-pyrrolo[3,4-b]pyridine and the
like), pyrazolopyridine (e.g., 1H-pyrazolo[3,4-b]pyridine,
1H-pyrazolo[4,3-b]pyridine and the like), imidazopyridine (e.g.,
1H-imidazo[4,5-b]pyridine and the like), pyrrolopyrimidine (e.g.,
1H-pyrrolo[2,3-d]pyrimidine, 1H-pyrrolo[3,2-d]pyrimidine,
1H-pyrrolo[3,4-d]pyrimidine and the like), pyrazolopyrimidine
(e.g., 1H-pyrazolo[3 ,4-d]pyrimidine, pyrazolo[1,5-a]pyrimidine,
1H-pyrazolo[4,3-d]pyrimidine and the like), imidazopyrimidine
(e.g., imidazo[1,2-a]pyrimidine, 1H-imidazo[4,5-d]pyrimidine and
the like), pyrrolotriazine (e.g., pyrrolo[1,2-a]-1,3,5-triazine,
pyrrolo[2,1-f]-1,2,4-triazine), pyrazolotriazine (e.g., pyrazolo[
1,5-a]- 1,3,5-triazine and the like), triazolopyridine (e.g., 1H-
1,2,3-triazolo[4,5-b]pyridine and the like), triazolopyrimidine
(e.g., 1,2,4-triazolo[1,5-a]pyrimidine,
1,2,4-triazolo[4,3-a]pyrimidine, 1H-1,2,3-triazolo[4,5-d]pyrimidine
and the like), cinnoline, quinazoline, quinoline, pyridopyridazine
(e.g., pyrido[2,3-c]pyridazine and the like), pyridopyrazine (e.g.,
pyrido[2,3-b]pyrazine and the like), pyridopyrimidine (e.g.,
pyrido[2,3-d]pyrimidine, pyrido[3,2-d]pyrimidine and the like),
pyrimidopyrimidine (e.g., pyrimido[4,5-d]pyrimidine,
pyrimido[5,4-d]pyrimidine and the like), pyrazinopyrimidine (e.g.,
pyrazino[2,3-d]pyrimidine and the like), naphthyridine (e.g.,
1,8-naphthyridine and the like), tetrazolopyrimidine (e.g.,
tetrazolo[ 1,5-a]pyrimidine and the like), thienopyridine (e.g.,
thieno[2,3-b]pyridine and the like), thienopyrimidinc (e.g.,
thieno[2,3-d]pyrimidine and the like), thiazolopyridine (e.g.,
thiazolo[4,5-b]pyridine, thiazolo[5,4-b]pyridine and the like),
thiazolopyrimidine (e.g., thiazolo[4,5-d]pyrimidine,
thiazolo[5,4-d]pyrimidine and the like), oxazolopyridine (e.g.,
oxazolo[4,5-b]pyridine, oxazolo[5,4-b]pyridine and the like),
oxazolopyrimidine (e.g., oxazolo[4,5-d]pyrimidine,
oxazolo[5,4-d]pyrimidine and the like), furopyridine (e.g.,
furo[2,3-bipyridine, furo[3,2-b]pyridine and the like),
furopyrimidine (e.g., furo[2,3-d]pyrimidine, furo[3,2-d]pyrimidine
and the like), 2,3-dihydropyrrolopyridine (e.g.,
2,3-dihydro-1H-pyrrolo[2,3-b]pyridine,
2,3-dihydro-1H-pyrrolo[3,2-b]pyridine and the like),
2,3-dihydropyrrolopyrimidine (e.g.,
2,3-dihydro-1H-pyrrolo[2,3-d]pyrimidi- ne,
2,3-dihydro-1H-pyrrolo[3,2-d]pyrimidine and the like),
5,6,7,8-tetrahydropyrido[2,3-d]pyrimidine,
5,6,7,8-tetrahydro-1,8-naphthy- ridine, 5,6,7,8-tetrahydroquinoline
and the like. When these rings form a hydrogenated aromatic ring,
the carbon atom in the ring may be carbonyl and includes, for
example, 2,3-dihydro-2-oxopyrrolopyridine,
2,3-dihydro-2,3-dioxopyrrolopyridine,
7,8-dihydro-7-oxo-1,8-naphthyridine- , 5,6,7,8-tetrahydro-7-oxo-
1,8-naphthyridine and the like.
[0163] These rings may be substituted by a substituent such as
halogen, alkyl, alkoxy, aralkyl, haloalkyl, nitro, amino,
alkylamino, cyano, formyl, acyl, aminoalkyl, mono- or
dialkylaminoalkyl, azide, carboxy, alkoxycarbonyl, carbamoyl,
alkylcarbamoyl, alkoxyalkyl (e.g., methoxymethyl, methoxyethyl,
methoxypropyl, ethoxymethyl, ethoxyethyl, ethoxypropyl and the
like), optionally substituted hydrazino and the like.
[0164] As used herein, the substituent of the optionally
substituted hydrazino includes alkyl, aralkyl, nitro, cyano and the
like, wherein alkyl and aralkyl are as defined for R and R.sup.1
and exemplified by methyl hydrazino, ethyl hydrazino, benzyl
hydrazino and the like.
[0165] In the present specification, each symbol of the formula
(II) is defined as follows.
[0166] The linear or branched alkyl having 1 to 6 carbon atoms at
R.sup.13, R.sup.14, R.sup.15 and R.sup.16 is methyl, ethyl, propyl,
isopropyl, butyl, isobutyl, sec-butyl, tert-butyl, pentyl, hexyl
and the like.
[0167] Aryl at R.sup.14 and R.sup.15 is phenyl, naphthyl and the
like.
[0168] Aralkyl at R.sup.14 and R.sup.15 is as defined for R and
R.sup.1.
[0169] Alkylene having 4 or less carbon atoms, which is formed by
R.sup.13 and R.sup.14 directly bonded to each other, is methylene,
ethylene, trimethylene, propylene, tetramethylene and the like.
[0170] Alkyl having 1 to 10 carbon atoms, which substitutes
alkylene having 4 or less carbon atoms formed by R.sup.13 and
R.sup.14 directly bonded to each other, is linear or branched alkyl
having 1 to 10 carbon atoms. Examples thereof include methyl,
ethyl, propyl, isopropyl, butyl, isobutyl, sec-butyl, tert-butyl,
pentyl, hexyl, heptyl, octyl, nonyl, decyl and the like.
[0171] Alkyl having 1 to 6 carbon atoms which substitutes ethylene
and trimethylene formed by R.sup.13 and R.sup.14 directly bonded to
each other is linear or branched alkyl having 1 to 6 carbon atoms,
which is the same as those for R.sup.13.
[0172] The heterocycle formed by R.sup.14 and R.sup.15 directly or
via oxygen atom bonded together with the adjacent nitrogen atom is
pyrrolidino, piperidino, morpholino, homopiperidino, homomorpholino
and the like.
[0173] Alkylene having 2 to 4 carbon atoms formed by R.sup.17 and
R.sup.18 directly bonded to each other is ethylene, trimethylene,
propylene, tetramethylene and the like.
[0174] Alkylene having 2 to 6 carbon atoms at Alk is ethylene,
trimethylene, propylene, tetramethylene, pentamethylene,
hexamethylene and the like.
[0175] Alkyl having 1 to 6 carbon atoms and alkyl having 1 to 10
carbon atoms, which are the substituents of alkylene having 2 to 6
carbon atoms at Alk, are as defined for R.sup.13.
[0176] Aryl and aralkyl, which are the substituents of alkylene
having 2 to 6 carbon atoms at Alk, are as defined for R.sup.14.
[0177] The compound to be used as the Rho kinase inhibitor of the
present invention is, for example, a compound of the formula (I),
which is exemplified by the following compounds.
[0178] (1) 4-(2-pyridylcarbamoyl)piperidine
[0179] (2) 1-benzyloxycarbonyl-4-(4-pyridylcarbamoyl)piperidine
[0180] (3) 1-benzoyl-4-(4-pyridylcarbamoyl)piperidine
[0181] (4) 1-propyl-4-(4-pyridylcarbamoyl)piperidine
[0182] (5)
[3-(2-(2-thienylmethyl)phenoxy)-2-hydroxypropyl]-4-(4-pyridylca-
rbamoyl)piperidine
[0183] (6) 4-(4-pyridylcarbamoyl)piperidine
[0184] (7)
1-benzyl-4-(4-pyridylcarbamoyl)-1,2,5,6-tetrahydropyridine
[0185] (8) 3-(4-pyridylcarbamoyl)piperidine
[0186] (9) 1-benzyl-3-(4-pyridylcarbamoyl)piperidine
[0187]
(10)1-(2-(4-benzyloxyphenoxy0ethyl)-4-(N-(2-pyridyl)-N-benzylcarbar-
moyl]pyridine
[0188] (11) 1-formyl-4-(4-pyridylcarbamoyl)piperidine
[0189] (12) 4-(3-pyridylcarbamoyl)piperidine
[0190] (13) 1-isopropyl-4-(4-pyridylcarbamoyl)piperidine
[0191] (14) 1-methyl-4-(4-pyridylcarbamoyl)piperidine
[0192] (15) 1-hexyl-4-(4-pyridylcarbamoyl)piperidine
[0193] (16) 1-benzyl-4-(4-pyridylcarbamoyl)piperidine
[0194] (17) 1-(2-phenylethyl)-4-(4-pyridylcarbamoyl)piperidine
[0195] (18)
1-(2-(4-methoxyphenyl)ethyl)-4-(4-pyridylcarbamoyl)piperidine
[0196] (19)
1-(2-(4-methoxyphenyl)ethyl)-4-(2-pyridylcarbamoyl)piperidine
[0197] (20)
1-(2-(4-chlorophenyl)ethyl)-4-(4-pyridylcarbamoyl)piperidine
[0198] (21) 1-diphenylmethyl4-(2-pyridylcarbamoyl)piperidine
[0199] (22)
1-[2-(4-(5-methyl-3-oxo-2,3,4,5-tetrahydropyridazin-6-yl)pheny-
l)ethyl]-4-(2-pyridylcarbamoyl)piperidine
[0200] (23)
1-(4-(4,5-dihydro-2-furyl)phenyl)-4-(4-pyridylcarbamoyl)piperi-
dine
[0201] (24) 1-(2-nitrophenyl)-4-(4-pyridylcarbamoyl)piperidine
[0202] (25) 1-(2-aminophenyl)-4-(4-pyridylcarbamoyl)piperidine
[0203] (26) 1-nicotinoyl-4-(4-pyridylcarbamoyl)piperidine
[0204] (27) 1-isonicotinoyl-4-(4-pyridylcarbamoyl)piperidine
[0205] (28)
1-(3,4,5-trimethoxybenzoyl)-4-(4-pyridylcarbamoyl)piperidine
[0206] (29) 1-acetyl-4-(4-pyridylcarbamoyl)piperidine
[0207] (30)
1-(3-(4-fluorobenzoyl)propyl)-4-(4-pyridylcarbamoyl)piperidine
[0208] (31)
1-(3-(4-fluorobenzoyl)propyl)-4-(2-pyridylcarbamoyl)piperidine
[0209] (32)
1-(1-(4-hydroxybenzoyl)ethyl)-4-(2-pyridylcarbamoyl)piperidine
[0210] (33)
1-(1-(4-benzyloxybenzoyl)ethyl)-4-(2-pyridylcarbamoyl)piperidi-
ne
[0211] (34)
1-(2-(4-hydroxyphenoxy)ethyl)-4-(2-pyridylcarbamoyl)piperidine
[0212] (35)
1-(4-(4-fluorophenyl)-4-hydroxybutyl)-4-(4-pyridylcarbamoyl)pi-
peridine
[0213] (36)
1-(1-methyl-2-(4-hydroxyphenyl)-2-hydroxyethyl)-4-(2-pyridylca-
rbamoyl)piperidine
[0214] (37) 1-cinnamyl-4-(2-pyridylcarbamoyl)piperidine
[0215] (38)
1-(2-hydroxy-3-phenoxypropyl)-4-(4-pyridylcarbamoyl)piperidine
[0216] (39)
1-(2-hydroxy-3-phenoxypropyl)-4-(3-pyridylcarbamoyl)piperidine
[0217] (40)
1-(2-hydroxy-3-phenoxypropyl)-4-(2-pyridylcarbamoyl)piperidine
[0218] (41)
1-(2-phenylethyl)-4-[N-(2-pyridyl)-N-(2-(N,N-dimethylamino)eth-
yl)carbamoyl]piperidine
[0219] (42)
1-benzyloxycarbonyl-4-(2-pyridylcarbamoyl)piperidine
[0220] (43)
1-(3-chlorophenyl)carbamoyl-4-(4-pyridylcarbamoyl)piperidine
[0221] (44) 1-[N-(2-pyridyl)-N-(2-( N,
N-dimethylamino)ethyl)carbamoyl]pip- eridine
[0222] (45)
1-methyl-4-(4-pyridylcarbamoyl)-1,2,5,6-tetrahydropyridine
[0223] (46) 1-nicotinoyl-3-(4-pyridylcarbamoyl)piperidine
[0224] (47)
1-[2-(4-fluorobenzoyl)ethyl]-4-(4-pyridylcarbamoyl)piperidine
[0225] (48)
1-(6-chloro-2-methylimidazo[1,2-a]pyridine-3-carbonyl)-4-(4-py-
ridylcarbamoyl)piperidine
[0226] (49) 1-(4-nitrobenzyl)-4-(4-pyridylcarbamoyl)piperidine
[0227] (50) 1-hexyl-4-(4-pyridylcarbamoyl)piperidine
[0228] (51)
1-benzyloxycarbonyl-4-(2-chloro-4-pyridylcarbamoyl)piperidine
[0229] (52) 4-(2-chloro-4-pyridylcarbamoyl)piperidine
[0230] (53)
1-(2-chloronicotinoyl)4-(4-pyridylcarbamoyl)piperidine
[0231] (54) 3-(2-chloro-4-pyridylcarbamoyl)piperidine
[0232] (55)
1-(4-phthalimidobutyl)-4-(4-pyridylcarbamoyl)piperidine
[0233] (56)
1-(3,5-di-tert-butyl-4-hydroxycinnamoyl)-4-(4-pyridylcarbamoyl-
)piperidine
[0234] (57) 1-carbamoylmethyl-4-(4-pyridylcarbamoyl)piperidine
[0235] (58)
1-benzyloxycarbonyl-4-(5-nitro-2-pyridylcarbamoyl)piperidine
[0236] (59) 4-(5-nitro-2-pyridylcarbamoyl)piperidine
[0237] (60)
trans-4-benzyloxycarboxamidomethyl-1-(4-pyridylcarbamoyl)cyclo-
hexane
[0238] (61)
trans-4-aminomethyl-1-(4-pyridylcarbamoyl)cyclohexane
[0239] (62)
trans-4-formamidomethyl-1-(4-pyridylcarbamoyl)cyclohexane
[0240] (63)
trans-4-dimethylaminomethyl-1-(4-pyridylcarbamoyl)cyclohexane
[0241] (64)
N-benzylidene-trans-(4-pyridylcarbamoyl)cyclohexylmethylamine
[0242] (65)
trans-4-benzylaminomethyl-1-(4-pyridylcarbamoyl)cyclohexane
[0243] (66)
trans-4-isopropylaminomethyl-1-(4-pyridylcarbamoyl)cyclohexane
[0244] (67) trans-4-nicotinoylaminomethyl- 1-(4-pyridylcarbamoyl
cyclohexane
[0245] (68)
trans-4-cyclohexylaminomethyl-1-(4-pyridylcarbamoyl)cyclohexan-
e
[0246] (69)
trans-4-benzyloxycarboxamide-1-(4-pyridylcarbamoyl)cyclohexane
[0247] (70) trans-4-amino-1-(4-pyridylcarbamoyl)cyclohexane
[0248] (71)
trans-4-(1-aminoethyl)-1-(4-pyridylcarbamoyl)cyclohexane
[0249] (72)
trans-4-aminomethyl-cis-2-methyl-1-(4-pyridylcarbamoyl)cyclohe-
xane
[0250] (73)
(+)-trans-4-(1-benzyloxycarboxamidopropyl)-1-cyclohexanecarbox-
ylic acid
[0251] (74)
(+)-trans-4-(1-benzyloxycarboxamidopropyl)-1-(4-pyridylcarbamo-
yl)cyclohexane
[0252] (75)
(-)-trans-4-(1-benzyloxycarboxamidpropyl)-1-(4-pyridylcarbamoy-
l)cyclohexane
[0253] (76)
(+)-trans-4-(1-aminopropyl)-1-(4-pyridylcarbamoyl)cyclohexane
[0254] (77)
(-)-trans4-(1-aminopropyl)-1-(4-pyridylcarbamoyl)cyclohexane
[0255] (78)
(-)-trans4-(1-benzyloxycarboxamidoethyl)-1-(4-pyridylcarbamoyl-
)cyclohexane
[0256] (79)
(+)-trans-4-(1-benzyloxycarboxamidoethyl)-1-(4-pyridylcarbamoy-
l)cyclohexane
[0257] (80)
(+)-trans-4-(1-aminoethyl)-1-(4-pyridylcarbamoyl)cyclohexane
[0258] (81)
(-)-trans4-(1-aminoethyl)-1-(4-pyridylcarbamoyl)cyclohexane
[0259] (82)
trans-4-(4-chlorobenzoyl)aminomethyl-1-(4-pyridylcarbamoyl)cyc-
lohexane
[0260] (83)
trans-4-aminomethyl-1-(2-pyridylcarbamoyl)cyclohexane
[0261] (84)
trans-4-benzyloxycarboxamidomethyl-1-(2-pyridylcarbamoyl)cyclo-
hexane
[0262] (85)
trans-4-methylaminomethyl-1-(4-pyridylcarbamoyl)cyclohexane
[0263] (86) trans-4-(
N-benzyl-N-methylamino)methyl-1-(4-pyridylcarbamoyl)-
cyclohexane
[0264] (87) trans-4-aminomethyl-N
-(3-pyridylcarbamoyl)cyclohexane
[0265] (88)
trans-4-aminomethyl-1-[(3-hydroxy-2-pyridyl)carbamoyl]cyclohex-
ane
[0266] (89)
trans-4-benzyloxycarboxamidomethyl-1-(3-pyridylcarbamoyl)cyclo-
hexane
[0267] (90)
trans-4-benzyloxycarboxamidomethyl-1-[(3-benzyloxy-2-pyridyl)c-
arbamoyl]cyclohexane
[0268] (91)
trans-4-phthalimidomethyl-1-(4-pyridylcarbamoyl)cyclohexane
[0269] (92)
trans-4-benzyloxycarboxamidomethyl-1-(3-methyl-4-pyridylcarbam-
oyl)cyclohexane
[0270] (93)
trans-4-aminomethyl-1-(3-methyl-4-pyridylcarbamoyl)cyclohexane
[0271] (94)
4-(trans-4-benzyloxycarboxamidomethylcyclohexylcarbonyl)amino--
2,6-dimethylpyridine-N-oxide
[0272] (95)
4-(trans-4-aminomethylcyclohexylcarbonyl)amino-2,6-dimethylpyr-
idine-N-oxide
[0273] (96)
trans-4-aminomethyl-1-(2-methyl-4-pyridylcarbamoyl)cyclohexane
[0274] (97)
trans-4-(1-benzyloxycarboxamidoethyl)-1-(4-pyridylcarbamoyl)cy-
clohexane
[0275] (98)
trans-4-(1-amino-1-methylethyl)-1-(4-pyridylcarbamoyl)cyclohex-
ane
[0276] (99)
trans-4-(2-aminoethyl)-1-(4-pyridylcarbamoyl)cyclohexane
[0277] (100)
trans-4-(2-amino-1-methylethyl)-1-(4-pyridylcarbamoyl)cyclohe-
xane
[0278] (101)
trans-4-(1-aminopropyl)-1-(4-pyridylcarbamoyl)cyclohexane
[0279] (102)
trans-4-aminomethyl-trans-1-methyl-1-(4-pyridylcarbamoyl)cycl-
ohexane
[0280] (103)
trans-4-benzylaminomethyl-cis-2-methyl-1-(4-pyridylcarbamoyl)-
cyclohexane
[0281] (104)
trans-4-(1-benzyloxycarboxamide-1-methylethyl)-1-(4-pyridylca-
rbamoyl)cyclohexane
[0282] (105) trans-4-benzyloxycarboxamidomethyl- 1-(
N-methyl-4-pyridylcarbamoyl)cyclohexane
[0283] (106)
trans-4-(1-acetamide-1-methylethyl)-1-(4-pyridylcarbamoyl)cyc-
lohexane
[0284] (107)
trans-N-(6-amino-4-pyrimidyl)-4-aminomethylcyclohexanecarboxa-
mide
[0285] (108)
trans-N-(1H-pyrrolo[2,3-b]pyridin-4-yl)-4-aminomethylcyclohex-
anecarboxamide
[0286] (109)
(+)-trans-N-(1H-pyrrolo[2,3-b]pyridin-4-yl)-4-(1-aminoethyl)c-
yclohexanecarboxamide
[0287] (110)
trans-N-(1H-pyrrolo[2,3-b]pyridin-4-yl)-4-(1-amino-1-methylet-
hyl)cyclohexanecarboxamide
[0288] (111)
trans-N-(1H-pyrazolo[3,4-b]pyridin-4-yl)-4-aminomethylcyclohe-
xanecarboxamide
[0289] (112)
(+)-trans-N-(1H-pyrazolo[3,4-b]pyridin-4-yl)-4-(1-aminoethyl)-
cyclohexanecarboxamide
[0290] (113)
trans-N-(1H-pyrazolo[3,4-b]pyridin-4-yl)-4-(1-amino-1-methyle-
thyl)cyclohexanecarboxamide
[0291] (114)
(+)-trans-N-(2-amino-4-pyridyl)-4-(1-aminoethyl)cyclohexaneca-
rboxamide
[0292] (115)
trans-N-(1H-pyrazolo[3,4-d]pyrimidin-4-yl)-4-aminomethylcyclo-
hexanecarboxamide
[0293] (116)
(+)-trans-N-(1H-pyrazolo[3,4-d]pyrimidin-4-yl)-4-(1-aminoethy-
l)cyclohexanecarboxamide
[0294] (117)
trans-N-(1H-pyrazolo[3,4-d]pyrimidin-4-yl)-4-(1-amino-1-methy-
lethyl)cyclohexanecarboxamide
[0295] (118)
trans-N-(4-pyrimidinyl)-4-aminomethylcyclohexanecarboxamide
[0296] (119)
trans-N-(3-amino-4-pyridyl)-4-aminomethylcyclohexanecarboxami-
de
[0297] (120)
trans-N-(7H-imidazo[4,5-d]pyrimidin-6-yl)-4-aminomethylcycloh-
exanecarboxamide
[0298] (121)
trans-N-(3H-1,2,3-triazolo[4,5-d]pyrimidin-7-yl)-4-aminomethy-
lcyclohexanecarboxamide
[0299] (122)
trans-N-(1-benzyl-1H-pyrazolo[3,4-b]pyridin-4-yl)-4-aminometh-
ylcyclohexanecarboxamide
[0300] (123)
trans-N-(1H-5-pyrazolyl)-4-aminomethylcyclohexanecarboxamide
[0301] (124)
trans-N-(1H-pyrazolo[3,4-b]pyridin-4-yl)-4-aminomethylcyclohe-
xanecarboxamide
[0302] (125)
trans-N-(4-pyridazinyl)-4-aminomethylcyclohexanecarboxamide
[0303] (126)
trans-N-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)-4-aminomethylcycloh-
exanecarboxamide
[0304] (127)
trans-N-(2-amino-4-pyridyl)-4-aminomethylcyclohexanecarboxami-
de
[0305] (128)
trans-N-(thieno[2,3-d]pyrimidin-4-yl)-4-aminomethylcyclohexan-
ecarboxamide
[0306] (129)
trans-N-(5-methyl-1,2,4-triazolo[1,5-a]pyrimidin-7-yl)-4-amin-
omethylcyclohexanecarboxamide
[0307] (130)
trans-N-(3-cyano-5-methylpyrazolo[1,5-a]pyrimidin-7-yl)-4-ami-
nomethylcyclohexanecarboxamide
[0308] (131) trans-N-(1H-pyrazolo[3,4-b]pyridin-4-yl)-4-( l-amino-
1-methylethyl)cyclohexanecarboxamide
[0309] (132)
trans-N-(2-(1-pyrrolidinyl)-4-pyridyl)-4-aminomethylcyclohexa-
necarboxamide
[0310] (133)
trans-N-(2,6-diamino-4-pyrimidyl)-4-aminomethylcyclohexanecar-
boxamide
[0311] (134) (+)-trans-N-(7-methyl-
1,8-naphthyridin-4-yl)-4-(1-aminoethyl- )cyclohexanecarboxamide
[0312] (135)
trans-N-(1-benzyloxymethylpyrrolo[2,3-b]pyridin-4-yl)-4-amino-
methylcyclohexanecarboxamide
[0313] (136)
(+)-trans-N-(1-methylpyrrolo[2,3-b]pyridin-4-yl)-4-(1-aminoet-
hyl)cyclohexanecarboxamide
[0314] (137)
trans-N-benzyl-N-(2-benzylamino-4-pyridyl)-4-(1-amino-1-methy-
lethyl)cyclohexanecarboxamide
[0315] (138)
trans-N-(2-azide-4-pyridyl)-4-aminomethylcyclohexanecarboxami-
de
[0316] (139)
trans-N-(2,3-dihydro-1H-pyrrolo[2,3-b]pyridin-4-yl)-4-aminome-
thylcyclohexanecarboxamide
[0317] (140)
trans-N-(2,3-dihydro-1H-pyrrolo[2,3-b]pyridin-4-yl)-4-(1-amin-
o-1-methylethyl)cyclohexanecarboxamide
[0318] (141-1)
trans-N-(2-carboxy-4-pyridyl)-4-aminomethylcyclohexanecarbo-
xamide
[0319] (141-2)
(R)-(+)-trans-N-(3-bromo-1H-pyrrolo[2,3-b]pyridin-4-yl)-4-(-
1-aminoethyl)cyclohexanecarboxamide
[0320] (142)
trans-N-(1H-pyrrolo[2,3-b]pyridin-4-yl)-4-guanidinomethylcycl-
ohexanecarboxamide
[0321] (143)
trans-N-(1H-pyrazolo[3,4-b]pyridin-4-yl)-4-guanidinomethylcyc-
lohexanecarboxamide
[0322] (144)
trans-N-(4-pyridyl)-4-guanidinomethylcyclohexanecarboxamide
[0323] (145)
trans-N-(1-methylpyrrolo[2,3-b]pyridin-4-yl)-4-(guanidinometh-
yl)cyclohexanecarboxamide
[0324] (146)
trans-N-(1H-pyrrolo[2,3-b]pyridin-4-yl)-4-(2-imidazolin-2-yl)-
aminomethylcyclohexanecarboxanide
[0325] (147)
trans-N-(1-benzyloxymethylpyrrolo[2,3-b]pyridin-4-yl)-4-guani-
dinomethylcyclohexanecarboxamide
[0326] (148)
trans-N-(2-amino-4-pyridyl)-4-guanidinomethylcyclohexanecarbo-
xamide
[0327] (149)
trans-N-(1-benzyloxymethyl-1H-pyrrolo[2,3-b]pyridin-4-yl)-4-(-
2-imidazolin-2-yl) aminomethylcyclohexanecarboxamide
[0328] (150)
trans-N-(1H-pyrrolo[2,3-b]pyridin-4-yl)-4-(3-benzylguanidinom-
ethyl)cyclohexanecarboxamide
[0329] (151)
trans-N-(1H-pyrrolo[2,3-b]pyridin-4-yl)-4-(3-phenylguanidinom-
ethyl)cyclohexanecarboxamide
[0330] (152)
trans-N-(1H-pyrrolo[2,3-b]pyridin-4-yl)-4-(3-propylguanidinom-
ethyl)cyclohexanecarboxamide
[0331] (153)
trans-N-(1H-pyrrolo[2,3-b]pyridin-4-yl)-4-(3-octylguanidinome-
thyl)cyclohexanecarboxamide
[0332] (154)
trans-N-(1-benzyloxymethylpyrrolo[2,3-b]pyridin-4-yl)-4-(2-be-
nzyl-3-ethylguanidinomethyl)cyclohexanecarboxamide
[0333] (155)
trans-N-(1H-pyrrolo[2,3-b]pyridin-4-yl)-4-(imidazol-2-yl)amin-
omethylcyclohexanecarboxamide
[0334] (156)
trans-N-(1H-pyrrolo[2,3-b]pyridin-4-yl)-4-(thiazol-2-yl)amino-
methylcyclohexanecarboxamide
[0335] (157) (R)-(+)-N-(4-pyridyl)4-(1-aminoethyl)benzamide
[0336] (158) N-(4-pyridyl)-4-(1-amino-1-methylethyl)benzamide
[0337] (159) N-(4-pyridyl)-4-aminomethyl-2-benzyloxybenzamide
[0338] (160) N-(4-pyridyl)-4-aminomethyl-2-ethoxybenzamide
[0339] (161)
(R)-(-)-N-(4-pyridyl)-4-(1-aminoethyl)-3-nitrobenzamide
[0340] (162)
(R)-(-)-N-(4-pyridyl)-3-amino4-(1-aminoethyl)benzamide
[0341] (163)
(R)-(+)-N-(4-pyridyl)-4-(1-aminoethyl)-3-chlorobenzamide
[0342] (164) N-(4-pyridyl)-3-aminomethylbenzamide
[0343] (165)
(R)-(+)-N-(1H-pyrrolo[2,3-b]pyridin-4-yl)-4-(1-aminoethyl)ben-
zamide
[0344] (166)
(R)-(+)-N-(1H-pyrazolo[3,4-b]pyridin-4-yl)-4-(1-aminoethyl)be-
nzamide
[0345] (167)
N-(1H-pyrazolo[3,4-b]pyridin-4-yl)-4-guanidinomethylbenzamide
[0346] (168) N-(4-pyridyl)-4-guanidinomethylbenzamide
[0347] (169)
(R)-(+)-N-(4-pyridyl)4-(1-aminoethyl)-3-fluorobenzamide
[0348] (170) N-(4-pyridyl)-4-aminomethylbenzamide
[0349] (171) N-(4-pyridyl)-4-aminomethyl-2-hydroxybenzamide
[0350] (172) N-(4-pyridyl)-4-(2-aminoethyl)benzamide
[0351] (173) N-(4-pyridyl)-4-aminomethyl-3-nitrobenzamide
[0352] (174) N-(4-pyridyl)-3-amino-4-aminomethylbenzamide
[0353] (175) (S)-(-)-N-(4-pyridyl)-4-(1-aminoethyl)benzamide
[0354] (176) (S)-(-)-N-(4-pyridyl)-2-(1-aminoethyl)benzamide
[0355] (177)
(R)-(+)-N-(4-pyridyl)-4-(1-aminoethyl)-2-chlorobenzamide
[0356] (178)
(R)-(+)-N-(1H-pyrrolo[2,3-b]pyridin-4-yl)-4-(1-(3-propylguani-
dino)ethyl)benzamide
[0357] (179)
(R)-(-)-N-(1H-pyrrolo[2,3-b]pyridin-4-yl)-4-(1-aminoethyl)-3--
azidebenzamide
[0358] (180)
(R)-(+)-N-(4-pyridyl)-4-(1-aminoethyl)-2-nitrobenzamide
[0359] (181)
(R)-(-)-N-(4-pyridyl)-4-(1-aminoethyl)-3-ethoxybenzamide
[0360] (182)
(R)-(+)-N-(3-iodo-1H-pyrrolo[2,3-b]pyridin-4-yl)-4-(1-aminoet-
hyl)benzamide
[0361] (183)
(R)-(+)-N-(3-iodo-1H-pyrrolo[2,3-b]pyridin-4-yl)-4-(1-aminoet-
hyl)-3-azidebenzamide
[0362] (184)
(R)-(-)-N-(4-pyridyl)-4-(1-aminoethyl)-3-hydroxybenzamide
[0363] (185)
N-(1H-pyrazolo[3,4-b]pyridin-4-yl)-4-guanidinomethyl-3-nitrob-
enzamide
[0364] (186)
(R)-N-(1H-pyrazolo[3,4-b]pyridin-4-yl)-4-(1-guanidinoethyl)-3-
-nitrobenzamide
[0365] (187)
(R)-N-(1H-pyrazolo[3,4-b]pyridin-4-yl)-4-(1-aminoethyl)-2-nit-
robenzamide
[0366] (188)
N-(1H-pyrazolo[3,4-b]pyridin-4-yl)-4-guanidinobenzamide
[0367] (189)
(R)-N-(1H-pyrazolo[3,4-b]pyridin-4-yl)-4-(1-aminoethyl)-3-nit-
robenzamide
[0368] (190)
(R)-N-(1H-pyrazolo[3,4-b]pyridin-4-yl)-4-(1-guanidinoethyl)be-
nzamide
[0369] (191) N-(1H-pyrazolo[3,4-b]pyridin-4-yl)
-4-(1-amino-2-hydroxyethyl- )benzamide
[0370] (192)
N-(1H-pyrazolo[3,4-b]pyridin-4-yl)-4-aminomethyl-3-nitrobenza-
mide
[0371] (193)
N-(1H-pyrrolo[2,3-b]pyridin-4-yl)-4-piperidinecarboxamide
[0372] (194)
N-(1H-pyrazolo[2,4-b]pyridin-4-yl)-4-piperidinecarboxamide
[0373] (195)
N-(1H-pyrazolo[3,4-b]pyridin-4-yl)-1-aminoacetyl4-piperidinec-
arboxamide
[0374] (196)
N-(1-methoxymethyl-1H-pyrazolo[3,4-b]pyridin-4-yl)-4-piperidi-
necarboxamide
[0375] (197)
N-(2,3-dihydro-1H-pyrrolo[2,3-b]pyridin-4-yl)-4-piperidinecar-
boxamide
[0376] (198)
N-(1H-pyrrolo[2,3-b]pyridin-4-yl)-1-(2-phenylethyl)-4-piperid-
inecarboxamide
[0377] (199)
N-(1H-pyrrolo[2,3-b]pyridin-4-yl)-1-amidino-4-piperidinecarbo-
xamide
[0378] (200)
N-(1H-pyrrolo[2,3-b]pyridin-4-yl)-1-(3-phenylpropyl)-4-piperi-
dinecarboxamide
[0379] (201)
N-(1H-pyrrolo[2,3-b]pyridin-4-yl)-1-benzyl-4-piperidinecarbox-
amide
[0380] (202)
N-(1H-pyrazolo[3,4-b]pyridin-4-yl)-1-(2-phenylethyl)-4-piperi-
dinecarboxamide
[0381] (203)
N-(1H-pyrazolo[3,4-b]pyridin-4-yl)-1-(3-phenylpropyl)-4-piper-
idinecarboxamide
[0382] Preferred are compounds (80), (109), (110), (112), (115),
(142), (143), (144), (145), (153), (157), (163), (165), (166) and
(179).
[0383] The compound to be used as the Rho kinase inhibitor of the
present invention is, for example, a compound of the formula (II),
which is exemplified by the following compounds.
[0384] (204) 1-(5-isoquinolinesulfonyl)homopiperazine
[0385] (205) 1-(5-isoquinolinesulfonyl)-2-methylhomopiperazine
[0386] (206) 1-(5-isoquinolinesulfonyl)-3-methylhomopiperazine
[0387] (207) 1-(5-isoquinolinesulfonyl)-6-methylhomopiperazine
[0388] (208)
1-(5-isoquinolinesulfonyl)-2,3-dimethylhomopiperazine
[0389] (209)
1-(5-isoquinolinesulfonyl)-3,3-dimethylhomopiperazine
[0390] (210) 1-(5-isoquinolinesulfonyl)-3-ethylhomopiperazine
[0391] (211) 1-(5-isoquinolinesulfonyl)-3-propylhomopiperazine
[0392] (212)
1-(5-isoquinolinesulfonyl)-3-isobutylhomopiperazine
[0393] (213) 1-(5-isoquinolinesulfonyl)-3-phenylhomopiperazine
[0394] (214) 1-(5-isoquinolinesulfonyl)-3-benzylhomopiperazine
[0395] (215) 1-(5-isoquinolinesulfonyl)-6-ethylhomopiperazine
[0396] (216) 1-(5-isoquinolinesulfonyl)-6-propylhomopiperazine
[0397] (217) 1-(5-isoquinolinesulfonyl)-6-butylhomopiperazine
[0398] (218) 1-(5-isoquinolinesulfonyl)-6-pentylhomopiperazine
[0399] (219) 1-(5-isoquinolinesulfonyl)-6-hexylhomopiperazine
[0400] (220)1-(5-isoquinolinesulfonyl)-6-phenylhomopiperazine
[0401] (221) 1-(5-isoquinolinesulfonyl)-6-benzylhomopiperazine
[0402] (222) 1-(5-isoquinolinesulfonyl)-4-methylhomopiperazine
[0403] (223)1 -(5-isoquinolinesulfonyl)-4-ethylhomopiperazine
[0404] (224) 1-(5-isoquinolinesulfonyl)-4-propylhomopiperazine
[0405] (225) 1-(5-isoquinolinesulfonyl)-4-butylhomopiperazine
[0406] (226) 1-(5-isoquinolinesulfonyl)-4-hexylhomopiperazine
[0407] (227)
N-(2-aminoethyl)-1-chloro-5-isoquinolinesulfonamide
[0408] (228)
N-(4-aminoethyl)-1-chloro-5-isoquinolinesulfonamide
[0409] (229)
N-(2-amino-1-methylethyl)-1-chloro-5-isoquinolinesulfonamide
[0410] (230) N-(2-amino-1-methylpentyl)-1-chloro-5-isoquinoline
[0411] (231) N-(3-amino-2-methylbutyl)-
1-chloro-5-isoquinolinesulfonamide
[0412] (232)
N-(3-di-n-butylaminopropyl)-1-chloro-5-isoquinolinesulfonamid-
e
[0413] (233) N-(
N-cyclohexyl-N-methylaminoethyl)-1-chloro-5-isoquinolines-
ulfonamide
[0414] (234)
N-(2-guanidinoethyl)-1-chloro-5-isoquinolinesulfonamide
[0415] (235)
N-(2-guanidinobutyl)-5-chloro-5-isoquinolinesulfonamide
[0416] (236)
N-(2-guanidino-1-methylethyl)-1-chloro-5-isoquinolinesulfonam-
ide
[0417] (237)
N-(2-guanidinomethylpentyl)-1-chloro-5-isoquinolinesulfonamid-
e
[0418] (238)
N-(2-guanidino-3-methylbutyl)-1-chloro-5-isoquinolinesulfonam-
ide
[0419] (239)
N-(3-guanidino-2-methylpropyl)-1-chloro-5-isoquinolinesulfona-
mide
[0420] (240)
N-(4-guanidino-3-methylbutyl)-1-chloro-5-isoquinolinesulfonam-
ide
[0421] (241)
2-methyl-4-(1-chloro-5-isoquinolinesulfonyl)piperazine
[0422] (242)
2-ethyl-4-(1-chloro-5-isoquinolinesulfonyl)piperazine
[0423] (243)
2-isobutyl-4-(1-chloro-5-isoquinolinesulfonyl)piperazine
[0424] (244)
2,5-dimethyl-4-(1-chloro-5-isoquinolinesulfonyl)piperazine
[0425] (245)
1-methyl-4-(1-chloro-5-isoquinolinesulfonyl)piperazine
[0426] (246)
1-amidino-4-(1-chloro-5-isoquinolinesulfonyl)piperazine
[0427] (247)
1-amidino4-(1-chloro-5-isoquinolinesulfonyl)homopiperazine
[0428] (248)
1-amidino-3-methyl-4-(1-chloro-5-isoquinolinesulfonyl)piperaz-
ine
[0429] (249)
1-amidino-2,5-dimethyl-4-(1-chloro-5-isoquinolinesulfonyl)pip-
erazine
[0430] (250)
N-(2-aminoethyl)-1-hydroxy-5-isoquinolinesulfonamide
[0431] (251)
N-(4-aminobutyl)-1-hydroxy-5-isoquinolinesulfonamide
[0432] (252)
N-(2-amino-1-methylethyl)-1-hydroxy-5-isoquinolinesulfonamide
[0433] (253)
N-(2-amino-1-methylheptyl)-1-hydroxy-5-isoquinolinesulfonamid-
e
[0434] (254)
N-(3-amino-2-methylbutyl)-1-hydroxy-5-isoquinolinesulfonamide
[0435] (255)
N-[3-(N,N-dibutylamino)propyl]-1-hydroxy-5-isoquinolinesulfon-
amide
[0436] (256)
N-[2-(N-cyclohexyl-N-methylamino)ethyl]-1-hydroxy-5-isoquinol-
inesulfonamide
[0437] (257)
N-(2-guanidinoethyl)-1-hydroxy-5-isoquinolinesulfonamide
[0438] (258)
N-(4-guanidinobutyl)-1-hydroxy-5-isoquinolinesulfonamide
[0439] (259)
N-(2-guanidino-1)-methylethyl)-1-hydroxy-5-isoquinolinesulfon-
amide
[0440] (260)
N-(1-guanidinomethylpentyl)-1-hydroxy-5-isoquinolinesulfonami-
de
[0441] (261)
N-(2-guanidino-3-methylbutyl)-1-hydroxy-5-isoquinolinesulfona-
mide
[0442] (262)
N-(3-guanidino-2-methylpropyl)-1-hydroxy-5-isoquinolinesulfon-
amide
[0443] (263)
N-(4-guanidino-3-methylbutyl)-1-hydroxy-5-isoquinolinesulfona-
mide
[0444] (264)
2-methyl-4-(1-hydroxy-5-isoquinolinesulfonyl)piperazine
[0445] (265)
2-ethyl-4-(1-hydroxy-5-isoquinolinesulfonyl)piperazine
[0446] (266)
2-isobutyl-4-(1-hydroxy-5-isoquinolinesulfonyl)piperazine
[0447] (267)
2,5-dimethyl-4-(1-hydroxy-5-isoquinolinesulfonyl)piperazine
[0448] (268)
1-methyl-4-(1-hydroxy-5-isoquinolinesulfonyl)piperazine
[0449] (269)
1-amidino-4-(1-hydroxy-5-isoquinolinesulfonyl)piperazine
[0450] (270)
1-amidino-4-(1-hydroxy-5-isoquinolinesulfonyl)homopiperazine
[0451] (271)
1-amidino-3-methyl-4-(1-hydroxy-5-isoquinolinesulfonyl)pipera-
zine
[0452] (272)
1-amidino-2,5-dimethyl-4-(1-hydroxy-5-isoquinolinesulfonyl)pi-
perazine
[0453] (273)
N-(2-methylaminoethyl)-1-chloro-5-isoquinolinesulfonamide
[0454] (274)
N-(2-ethylaminoethyl)-1-chloro-5-isoquinolinesulfonamide
[0455] (275)
N-(2-propylaminoethyl)-1-chloro-5-isoquinolinesulfonamide
[0456] (276)
N-(2-butylaminoethyl)-1-chloro-5-isoquinolinesulfonamide
[0457] (277)
N-(2-hexylaminoethyl)-1-chloro-5-isoquinolinesulfonamide
[0458] (278) 1-(1-chloro-5-isoquinolinesulfonyl)piperazine
[0459] (279) 1-(1-chloro-5-isoquinolinesulfonyl)homopiperazine
[0460] (280)
N-(2-methylaminoethyl)-1-hydroxy-5-isoquinolinesulfonamide
[0461] (281)
N-(2-ethylaminoethyl)-1-hydroxy-5-isoquinolinesulfonamide
[0462] (282)
N-(2-propylaminoethyl)-1-hydroxy-5-isoquinolinesulfonamide
[0463] (283)
N-(2-butylaminoethyl)-1-hydroxy-5-isoquinolinesulfonamide
[0464] (284)
N-(2-hexylaminoethyl)-1-hydroxy-5-isoquinolinesulfonamide
[0465] (285) 1-(1-hydroxy-5-isoquinolinesulfonyl)piperazine
[0466] (286) 1-(1-hydroxy-5-isoquinolinesulfonyl)homopiperazine
[0467] (287) 1-(5-isoquinolinesulfonyl)-4-methylpiperazine
[0468] (288) 1-(5-isoquinolinesulfonyl)-4-n-hexylpiperazine
[0469] (289) 1-(5-isoquinolinesulfonyl)-4-cinnamylpiperazine
[0470] (290) 1-(5-isoquinolinesulfonyl)piperazine
[0471] (291) N-(2-aminoethyl)-5-isoquinolinesulfonamide
[0472] (292) N-(4-aminobutyl)-5-isoquinolinesulfonamide
[0473] (293)
N-(3-di-n-butylaminopropyl)-5-isoquinolinesulfonamide
[0474] (294) 1-(5-isoquinolinesulfonyl)-3-methylpiperazine
[0475] (295) 1-(5-isoquinolinesulfonyl)-3-isobutylpiperazine
[0476] (296) 1-(5-isoquinolinesulfonyl)-2,5-dimethylpiperazine
[0477] (297)
N-(3-guanidino-2-phenylpropyl)-5-isoquinolinesulfonamide
[0478] (298)
N-(6-guanidino-1-methylheptyl)-5-isoquinolinesulfonamide
[0479] (299)
2-[2-(5-isoquinolinesulfonamide)ethylamino]-2-imidazoline
[0480] (300) 2-amidino-1-(5-isoquinolinesulfonyl)piperazine
[0481] (301)
4-amidino-2,5-dimethyl-1-(5-isoquinolinesulfonyl)piperazine
[0482] (302) 4-amidino-1-(5-isoquinolinesulfonyl)homopiperazine
[0483] (303)
4-(N1,N2-dimethylamidino)-1-(5-isoquinolinesulfonyl)piperazin-
e
[0484] (304)
4-amidino-3-butyl-1-(5-isoquinolinesulfonyl)piperazine
[0485] (305) 4-hexyl-1-(5-isoquinolinesulfonyl)ethylenediamine
[0486] (306) N-(4-guanidinobutyl)-5-isoquinolinesulfonamide
[0487] (307) N-(2-guanidinoethyl)-5-isoquinolinesulfonamide
[0488] (308) 1-(5-isoquinolinesulfonyl)-2-methylpiperazine
[0489] Preferred are compounds (204) and (308).
[0490] The compound to be used as the Rho kinase inhibitor of the
present invention may be a pharmaceutically acceptable acid
addition salt. The acid is exemplified by inorganic acid such as
hydrochloric acid, hydrobromic acid, sulfuric acid and the like and
organic acid such as methanesulfonic acid, fumaric acid, maleic
acid, mandelic acid, citric acid, tartaric acid, salicylic acid and
the like. The compound having a carboxyl group can be converted to
a salt with a metal such as sodium, potassium, calcium, magnesium,
aluminum and the like or a salt with amino acid such as lysine and
the like. In addition, their monohydrate, dihydrates, 1/2 hydrates,
1/3 hydrates, 1/4 hydrates, 2/3 hydrates, 3/2 hydrates and the like
are also encompassed in the present invention.
[0491] The compound of the formula (I) can be synthesized according
to the method disclosed in Japanese Patent Unexamined Publication
No. 62-89679, Japanese Patent Unexamined Publication No. 3-218356,
Japanese Patent Unexamined Publication No. 5-194401, Japanese
Patent Unexamined Publication No. 6-41080, WO95/28387 and the
like.
[0492] The compound of the formula (II) can be synthesized
according to the method disclosed in Japanese Patent Unexamined
Publication No. 57-156463, Japanese Patent Unexamined Publication
No. 57-200366, Japanese Patent Unexamined Publication No.
58-121278, Japanese Patent Unexamined Publication No. 58-121279,
Japanese Patent Unexamined Publication No. 59-93054, Japanese
Patent Unexamined Publication No. 60-81168, Japanese Patent
Unexamined Publication No. 61-152658, Japanese Patent Unexamined
Publication No. 61-227581, Japanese Patent Unexamined Publication
No. 62-103066, U.S. Pat. No. 4,678,783 and the like.
[0493] Of the compounds of the formula (I), a compound wherein Ra
is a group of the formula (c) and Rc is Rc', namely, an amide
compound of the formula (III) 10
[0494] wherein Rc' is an optionally substituted heterocycle
containing nitrogen of the above-mentioned Rc except pyridine, and
other symbols are as defined above, is a novel compound which can
be synthesized by the following methods.
[0495] Method 1
[0496] A compound of the formula (IV)
Rc'-NH-Rb (IV)
[0497] wherein each symbol is as defined above, and a compound of
the formula (V) 11
[0498] wherein each symbol is as defined above, or a reactive
derivative thereof are reacted to give the compound. The reactive
derivative of carboxylic acid compound is exemplified by acid
halide, ester, acid anhydride, mixed acid anhydride and the
like.
[0499] This reaction beneficially proceeds by stirring in the
presence of a solvent inert to the reaction, such as
tetrahydrofuran, dioxane, chloroform, dichloromethane,
dimethylformamide, benzene, toluene, ethanol and the like. Water,
alcohol or acid liberated during the reaction is removed from the
reaction mixture by a method known in the pertinent field, such as
azeotropic distillation, forming a complex, converting to salt and
the like.
[0500] Method 2
[0501] Of the compounds of the formula (III), a compound wherein L
has a substituent other than hydrogen can be produced by reacting a
compound wherein L is hydrogen, with a compound of the formula
(VI)
L.sup.1-M (VI)
[0502] wherein L.sup.1 is, of the aforementioned L, a substituent
other than hydrogen and M is a reactive atom, according to
N-alkylation or N-acylation known in this field.
[0503] Method 3
[0504] Of the compounds of the formula (III), a compound wherein L
is alkyl or has a substituent having the formula (i) can be
produced by reductive amination reaction of a compound wherein L is
hydrogen and a compound of the formula (VII)
L.sup.2.dbd.C.dbd.O (VII)
[0505] wherein L.sup.2 is a group that can be converted to alkyl or
a group of the formula (i), by reductive amination reaction.
[0506] Method 4
[0507] Of the compounds of the formula (III), a compound wherein L
is a group of the formula (1) 12
[0508] wherein Q.sup.1 is as defined above and W.sup.1 is
hydroxytrimethylene from among the substituents at W, can be
produced by reacting a compound of the formula (III) wherein L is
hydrogen and a compound of the formula (VIII) 13
[0509] wherein Q.sup.1 is as defined above.
[0510] The reaction advantageously proceeds in a suitable solvent
which does not influence the reaction, such as alcohol (e.g.,
methanol, ethanol, 2-propanol and the like), aliphatic or alicyclic
ketone (e.g., 2-propanone, 2-butanone, cyclohexane and the like)
and the like. Addition of a suitable base such as alkali metal
carbonate, hydrogencarbonate and the like enables acceleration of
the reaction rate. The reaction temperature is rather elevating,
which is preferably refluxing temperature of the reaction
mixture.
[0511] Method 5
[0512] Of the compounds of the formula (III), a compound wherein L
is hydrogen can be produced from a compound of the formula (III-a)
14
[0513] wherein B.sup.1 is alkoxy or aralkyloxy, from among the
aforementioned substituents B, and other symbols are as defined
above.
[0514] Of the compounds (III-a), a compound wherein B.sup.1 is
alkoxy is stirred in a suitable organic solvent which does not
influence the reaction, such as alcohol (e.g., methanol, ethanol,
2-propanol and the like) and ether (e.g., tetrahydrofuran and the
like) in the presence of a suitable base, such as hydroxide of
alkali metal or alkaline earth metal, carbonate or
hydrogencarbonate (e.g., sodium hydroxide, potassium carbonate,
sodium hydrogencarbonate and the like) and heated as necessary to
give a compound of the formula (III) wherein L is hydrogen.
[0515] Of the compounds (III-a), a compound wherein B.sup.1 is
aralkyloxy is subjected to reductive decomposition reaction in a
suitable organic solvent which does not influence the reaction in
the presence of a suitable catalyst such as palladium carbon and
the like using a hydrogen source of hydrogen, hydrazine, formic
acid, ammonium formate and the like at normal temperature or under
pressurization where necessary.
[0516] Moreover, a compound (III-a) is stirred in 5-35%, preferably
15-30%, acetic acid in the presence of hydrogen bromide, whereby
the compound can be converted. A compound of the formula (III-b)
15
[0517] wherein Y.sup.1 is methylene, from among the aforementioned
substituents Y, and other symbols are as defined above, is
subjected to catalytic hydrogenation decomposition reaction wherein
the compound is stirred in a suitable organic solvent which does
not influence the reaction in the presence of a suitable catalyst
such as palladium carbon and the like under hydrogen to give a
compound of the formula (III) wherein L is hydrogen.
[0518] The compound of the formula (III) thus obtained can be
separated from the reaction mixture and purified by a method known
in the field of art, such as recrystallization, chromatography and
the like.
[0519] In addition, the compound of the formula (III) can form a
pharmaceutically acceptable salt by a conventional method. The acid
to be used for forming a salt can be appropriately selected from
inorganic acids such as hydrochloric acid, hydrobromic acid,
sulfuric acid and the like, organic acids such as methanesulfonic
acid, fumaric acid, maleic acid, mandelic acid, citric acid,
tartaric acid, salicylic acid and the like, amino acids such as
lysine and the like, and metal such as sodium, potassium, calcium,
magnesium, aluminum and the like. These acid addition salts can be
converted to a corresponding free base by the reaction with alkali
such as sodium hydroxide, potassium hydroxide and the like
according to a known method. The salts can be also converted to
quaternary ammonium.
[0520] The compound of the formula (III) may exist as optical
isomer, racemate thereof or cis-trans isomer, all of which are
encompassed in the present invention. These isomers can be isolated
by a conventional method or produced by using various starting
compounds.
[0521] When the Rho kinase inhibitor of the present invention is
used as a pharmaceutical agent, particularly as a therapeutic agent
of hypertension, a therapeutic agent of angina pectoris, a
suppressive agent of cerebrovascular contraction, a therapeutic
agent of asthma, a therapeutic agent of peripheral circulation
disorder, a prophylactic agent of immature birth, a therapeutic
agent of arteriosclerosis, an anti-cancer drug, an
anti-inflammatory agent, an immunosuppressant, a therapeutic agent
of autoimmune disease, a contraceptive, a prophylactic agent of
digestive tract infection, an anti-AIDS drug, a therapeutic agent
of osteoporosis, a therapeutic agent of retinopathy or a brain
function improving drug, it can be prepared as a general
pharmaceutical agent. For example, the Rho kinase inhibitor of the
present invention is mixed with a pharmaceutically acceptable
carrier (e.g., excipient, binder, disintegrator, corrective,
corrigent, emulsifier, diluent, solubilizer and the like) to give a
pharmaceutical composition or a pharmaceutical preparation in the
form of tablet, pill, powder, granule, capsule, troche, syrup,
liquid, emulsion, suspension, injection (e.g., liquid, suspension
and the like), suppository, inhalant, percutaneous absorber, eye
drop, eye ointment and the like in the form suitable for oral or
parenteral preparation.
[0522] When preparing a solid preparation, an additive such as
sucrose, lactose, cellulose sugar, D-mannitol, maltitol, dextran,
starches, agar, arginates, chitins, chitosans, pectines,
tragacanth, gum arabic, gelatins, collagens, casein, albumin,
calcium phosphate, sorbitol, glycine, carboxymethyl cellulose,
polyvinylpyrrolidone, hydroxypropylcellulose,
hydroxypropylmethylcellulose, glycerol, polyethyleneglycol, sodium
hydrogencarbonate, magnesium stearate, talc and the like are used.
Tablets can be applied with a typical coating, where necessary, to
give sugar coated tablets, enteric tablets, film-coated tablets,
two-layer tablets and multi-layer tablets.
[0523] When preparing a semi-solid preparation, animal and plant
fats and oils (e.g., olive oil, corn oil, castor oil and the like),
mineral fats and oils (e.g., petrolatum, white petrolatum, solid
paraffin and the like), wax (e.g., jojoba oil, carnauba wax, bee
wax and the like), partly or entirely synthesized glycerol fatty
acid esters (e.g., lauric acid, myristic acid, palmitic acid and
the like), and the like are used. Examples of commercially
available products of these include Witepsol (manufactured by
Dynamitnovel Ltd.), Farmazol (NOF Corporation) and the like.
[0524] When preparing a liquid preparation, an additive, such as
sodium chloride, glucose, sorbitol, glycerol, olive oil, propylene
glycol, ethyl alcohol and the like, is used. In particular, when
preparing an injection, a sterile aqueous solution such as
physiological saline, isotonizing liquid, oily liquid (e.g., sesame
oil and soybean oil) and the like is used. Where necessary, a
suitable suspending agent such as sodium carboxymethylcellulose,
nonionic surfactant, solubilizer (e.g., benzyl benzoate and benzyl
alcohol), and the like can be concurrently used. Moreover, when an
eye drop is prepared, an aqueous liquid or solution is used, which
is particularly a sterile injectable aqueous solution. The liquid
for an eye drop can appropriately contain various additives such as
buffer (preferred are borate buffer, acetate buffer, carbonate
buffer and the like for less irritation), isotonizing agent,
solubilizer, preservative, thickener, chelating agent, pH adjuster
(preferably, pH is generally adjusted to about 6-8.5) and
aromatic.
[0525] The content of the active ingredient in these preparation is
0.1-100 wt %, suitably 1-50 wt %, of the preparation. While subject
to variation depending on the condition, body weight, age and the
like of patient, in general, about 1-500 mg of the active
ingredient is orally administered daily for an adult in a single
dose or several doses.
EXAMPLES
[0526] The present invention is described in more detail in the
following by way of Examples, Formulation Examples and
pharmacological action, to which the present invention is not
limited.
[0527] In the following, the synthetic method of the novel compound
of the formula (III) of the present invention is described by
referring to examples.
Example 1
[0528] (a) N-Benzyloxycarbonylisonipecotyl chloride (5 g) was added
to a solution of
4-amino-1-tert-butoxycarbonyl-1H-pyrrolo[2,3-b]pyridine (3 g) and
diisopropylethyamine (2.16 g) in acetonitrile (40 ml) and the
mixture was stirred at room temperature for 2 hours. The reaction
mixture was poured into ice-water and extracted with chloroform.
The residue obtained by water washing, drying and then
concentration under reduced pressure was purified by silica gel
column chromatography to give 6.3 g of
N-(1-tert-butoxycarbonyl-1H-pyrrolo-[2,3-b]pyridin-4-yl)-1-benzyloxycarbo-
nyl-4-piperidinecarboxamide.
[0529] PMR(CDCl.sub.3): 1.67(9H, s), 1.79(2H, m), 1.95(2H, m),
2.53(1H, m), 2.89(2H, m), 4.29(2H, m), 5.15(2H, s), 6.48(1H,
d,J=4.4 Hz), 7.36(5H, m), 7.59(1H, br), 7.61(1H, d,J=4.4 Hz),
7.99(1H, d,J=5.4 Hz), 8.43(1H, d,J=5.4 Hz)
[0530] (b) N-(1
-tert-Butoxycarbonyl-1H-pyrrolo[2,3-b]pyridin-4-yl)-
1-benzyloxycarbonyl-4-piperidinecarboxamide (2 g) was dissolved in
methanol (30 ml) and 10% palladium carbon hydroxide (0.5 g) was
added for hydrogenation (normal pressure). After the completion of
the reaction, the catalyst was filtered off and the filtrate was
concentrated under reduced pressure to give 1.2 g of
N-(1-tert-butoxycarbonyl-1H-pyrrolo[2,3-
-b]pyridin-4-yl)-4-piperidinecarboxamide.
[0531] PMR(DMSO-d.sub.6): 1.59(9H, s), 1.83(2H, m), 2.01(2H, m),
2.89(2H, m), 3.01(1H, m), 3.32 (2H, m), 7.19(1H, d,J=4.4 Hz),
7.68(1H, d,J=4.4 Hz), 7.97(1H, d,J=5.4 Hz), 8.24(1H, d,J=5.4 Hz),
8.81(1H, br), 10.45(1H, s)
[0532] (c) Formic acid (10 ml) was added to
N-(1-tert-butoxycarbonyl-1H-py-
rrolo[2,3-b]pyridin-4-yl)-4-piperidinecarboxamide (1 g) and the
mixture was stirred at room temperature for 2 hours. The mixture
was neutralized with aqueous 1N sodium hydroxide solution and
extracted with chloroform. The crystals obtained by water washing,
drying and then concentration under reduced pressure were dissolved
in 15% hydrochloric acid-methanol solution (5 ml). The crystals
obtained by concentration of the resulting solution were
recrystallized from ethanol-ethyl acetate to give 650 mg of
N-(1H-pyrrolo[2,3-b]pyridin-4-yl)-4-piperidine-carboxamide mono
hydrochloride monohydrate, melting point 273.degree. C.
(decomposition).
[0533] PMR(DMSO-d.sub.6): 1.52(2H, m), 1.69(2H, m), 2.51(2H, m),
2.70(1H, m), 2.97(2H, m), 3.32(1H, br), 6.79(1H, d,J=3.4 Hz),
7.31(1H, d,J=3.4 Hz), 7.79(1H, d,J=5.4 Hz), 8.04(1H, d,J=5.4 Hz),
9.82(1H, s), 11.54(1H, br)
Example 2
[0534] (a) A solution of
N-(1-tert-butoxycarbonyl-1H-pyrrolo[2,3-b]pyridin-
-4-yl)-4-piperidinecarboxamide (0.6 g), phenetyl bromide (390 mg)
and potassium carbonate (290 mg) in dimethylformamide (10 ml) was
stirred at 80.degree. C. for 2 hours. The reaction mixture was
poured into ice water and extracted with chloroform. The residue
obtained by water washing, drying and then concentration under
reduced pressure was purified by silica gel column chromatography
to give 550 mg of N-(1 -tert-butoxycarbonyl- 1
H-pyrrolo[2,3-b]pyridin-4-yl)-1-(2-phenylethyl)-4-
-piperidinecarboxamide.
[0535] PMR(DMSO-d.sub.6): 1.59(9H, s), 1.66(2H, m), 1.80(2H, m),
1.98(2H, m), 2.50(2H, m), 2.56(1H, m), 2.74(2H, m), 3.01(2H, m),
7.05(1H, d,J=4.4Hz), 7.23(5H, m), 7.68(1H, d,J=4.4Hz),
7.97(1H,J=5.4 Hz), 8.23(1H, d,J=5.4 Hz), 10.03(1H, s)
[0536] (b) Formic acid (5 ml) was added to
N-(1-tert-butoxycarbonyl-1H-pyr-
rolo[2,3-b]pyridin-4-yl)-1-(2-phenylethyl)-4-piperidinecarboxamide
(550 mg) and the mixture was stirred at room temperature for 2
hours. The mixture was neutralized with aqueous 1N sodium hydroxide
solution and extracted with chloroform. The crystals obtained by
water washing, drying and then concentration under reduced pressure
were dissolved in 15% hydrochloric acid-methanol solution (1 ml).
The crystals obtained by concentration of the resulting solution
were recrystallized from ethanol-ethyl acetate to give 250 mg of
N-(1H-pyrrolo[2,3-b]pyridin-4-yl)-
-1-(2-phenylethyl)-4-piperidinecarboxamide dihydrochloride 1/4
hydrate, melting point 272.degree. C. (decomposition).
[0537] PMR(DMSO-d.sub.6/TMS): 2.00-2.19(4H, m), 2.93-3.41(7H, m),
3.63-3.68(2H, m), 7.22-7.37(5H, m), 7.50(1H, d,J=2.0 Hz), 7.56(1H,
t,J=2.0 Hz), 8.25(1H, d,J=6.8 Hz), 8.33(1H, d,J=6.8Hz), 10.86(1H,
br), 11.36(1H, s), 12.77(1H, br)
Example 3
[0538] (a) A solution of
N-(l-tert-butoxycarbonyl-1H-pyrrolo[2,3-b]pyridin-
-4-yl)-4-piperidinecarboxamide (500 mg), benzyl bromide (370 mg)
and potassium carbonate (300 mg) in dimethylformamide (10 ml) was
stirred at 80.degree. C. for 4 hours. The reaction mixture was
poured into ice-water and extracted with chloroform. The residue
obtained by water washing, drying and then concentration under
reduced pressure was purified by silica gel column chromatography
to give 300 mg of N-(1 -tert-butoxycarbonyl- 1
H-pyrrolo[2,3-b]pyridin-4-yl)- 1
-benzyl-4-piperidinecarboxamide.
[0539] PMR(CDCl.sub.3): 1.65(9H, s), 1.91(4H, m), 2.04(2H, m),
2.35(1H, m), 2.97(2H, m), 3.51(2H, s), 6.44(1H, d,J=3.9 Hz),
7.30(5H, m), 7.49(1H, br), 7.57(1H, d,J=3.9 Hz), 7.99(1H, d,J=5.4
Hz), 8.41(1H, d,J=5.4 Hz)
[0540] (b) Formic acid (4 ml) was added to
N-(1-tert-butoxycarbonyl-1H-pyr-
rolo[2,3-b]pyridin-4-yl)-1-benzyl-4-piperidinecarboxamide (300 mg)
and the mixture was stirred at room temperature for 1 hour. The
mixture was neutralized with aqueous 1N sodium hydroxide solution
and extracted with chloroform. The crystals obtained by water
washing, drying and then concentration under reduced pressure were
dissolved in 15% hydrochloric acid-methanol solution (1 ml). The
crystals obtained by concentration of the resulting solution were
recrystallized from ethanol-ethyl acetate to give 120 mg of
N-(1H-pyrrolo[2,3-b]pyridin-4-yl)-1-benzyl-4-piperidinecar-
boxamide dihydrochloride monohydrate, melting point 260.degree. C.
(decomposition).
[0541] PMR(DMSO-d.sub.6/TMS) : 2.00-2.15(4H, m), 2.92-2.98(2H, m),
3.13-3.19(1H, m), 3.36-3.43(2H, m), 4.32(2H, s), 7.55(1H, br),
7.63(2H, m), 8.20(1H, d,J=6.4 Hz), 8.31(1H, d,J=6.4 Hz), 10.76(1H,
br), 11.25(1H, br), 12.69(1H, br)
[0542] The following compounds can be obtained in the same manner
as in the above Examples.
Example 4
[0543] N-(1H-pyrazolo[3,4-b]pyridin-4-yl)-4-piperidinecarboxamide
dihydrochloride 3/2 hydrate, melting point 277.degree. C.
(decomposition)
Example 5
[0544] N-(1H-pyrazolo
[3,4-b]pyridin-4-yl)-1-aminoacetyl-4-piperidinecarbo- xamide
dihydrochloride 1/2 hydrate, melting point 264.degree. C.
(decomposition)
Example 6
[0545]
N-(1-methoxymethyl-1H-pyrazolo[3,4-b]pyridin-4-yl)-4-piperidinecarb-
oxamide monohydrate, melting point 240-241.degree. C.
Example 7
[0546]
N-(2,3-dihydro-1H-pyrrolo[2,3-b]pyridin-4-yl)-4-piperidinecarboxami-
de dihydrochloride 3/2 hydrate, melting point 235.degree. C.
(decomposition)
Example 8
[0547] N-( 1 H-pyrrolo[2,3-b]pyridin-4-yl)- 1
-amidino-4-piperidinecarboxa- mide dihydrochloride 5/4 hydrate,
melting point 246.degree. C. (decomposition)
Example 9
[0548]
N-(1H-pyrrolo[2,3-b]pyridin-4-yl)-1-(3-phenylpropyl)-4-piperidineca-
rboxamide dihydrochloride, melting point 276.degree. C.
(decomposition)
Example 10
[0549]
N-(1H-pyrazolo[3,4-b]pyridin-4-yl)-1-(2-phenylethyl)-4-piperidineca-
rboxamide dihydrochloride hydrate, melting point 259-26 1.degree.
C. (decomposition)
Example 11
[0550]
N-(1H-pyrazolo[3,4-b]pyridin-4-yl)-1-(3-phenylpropyl)-4-piperidinec-
arboxamide dihydrochloride 1/2 hydrate, melting point
240-244.degree. C. (decomposition)
[0551] A method for preparing the pharmaceutical preparation of the
present invention is explained in the following.
Formulation Example 1
[0552] Tablets
1 Inventive compound 10.0 mg Lactose 50.0 mg Corn starch 20.0 mg
Crystalline cellulose 29.7 mg Polyvinylpyrrolidone K30 5.0 mg Talc
5.0 mg Magnesium stearate 0.3 mg 120.0 mg
[0553] The inventive compound, lactose, corn starch and crystalline
cellulose were mixed, kneaded with polyvinylpyrrolidone K30 paste
solution and passed through a 20-mesh sieve for granulation. After
drying at 50.degree. C. for 2 hours, the granules were passed
through a 24-mesh sieve, and talc and magnesium stearate were
added. Using a .phi.7 mm punch, tablets weighing 120 mg per tablet
were prepared.
[0554] Formulation Example 2
[0555] Capsules
2 Inventive compound 10.0 mg Lactose 70.0 mg Corn starch 35.0 mg
Polyvinylpyrrolidone K30 2.0 mg Talc 2.7 mg Magnesium stearate 0.3
mg 120.0 mg
[0556] The inventive compound, lactose, corn starch and crystalline
cellulose were mixed, kneaded with polyvinylpyrrolidone K30 paste
solution and passed through a 20-mesh sieve for granulation. After
drying at 50.degree. C. for 2 hours, the granules were passed
through a 24-mesh sieve and talc and magnesium stearate were added.
The mixture was filled in hard capsules (No. 4) to give capsules
weighing 120 mg.
[0557] The pharmacological action of the pharmaceutical preparation
of the present invention is explained in the following by way of
experimental examples.
Experimental Example 1
[0558] Rho Kinase Inhibitory Action (Inhibition of Bovine Aorta
Thoracia Rho Kinase)
[0559] The Rho kinase was prepared from bovine aorta of thorax by
partial purification as in the following. The artery was minced and
homogenized with a 9-fold amount of 50 mM
Tris-hydroxymethylaminomethane (Tris) (pH=7.4), 1 mM
dithiothreitol, 1 mM EGTA, 1 mM EDTA, 100 .mu.M
p-amidinophenylmethylsulfonyl fluoride, 5 .mu.M E-64, 5 .mu.M
leupeptine and 5 .mu.M pepstatin A. The homogenate was centrifuged
(10,000.times.g, 30 minutes) to give supernatant. The supernatant
was adsorbed onto a hydroxyapatite column. The column was washed
with 0.2M phosphate buffer (pH=6.8). The standard product of Rho
kinase was eluted with 0.4M phosphate buffer (pH=6.8). The Rho
kinase was assayed as follows.
[0560] A reaction mixture (total amount 50 .mu.l) containing 50 mM
Tris, 1 mM EDTA, 5 mM MgCl.sub.2, 50 .mu.g/ml histone, 10 .mu.M
GTP.gamma.S, 100 .mu.g/ml Rho, 2 .mu.M [.sup.32P]ATP, the Rho
kinase (3 .mu.l) prepared in the above and the test compound was
reacted at 30.degree. C. for 5 minutes. The reaction was terminated
by the addition of 25% trichloroacetic acid (TCA) solution (1 ml)
and the mixture was stood at 4.degree. C. for 30 minutes. Then, the
mixture was filtered through a membrane filter (HAWP type,
Millipore), and the radioactivity of the filter was counted on a
liquid scintillation counter. The inhibitory action of the test
compound was calculated from the following formula based on the
comparison of the radioactivity with the sample without the test
compound (control). The results are shown in Table 1. 1 Inhibition
( % ) = [ cpm under control - cpm in the presence of test compound
cpm under control ] .times. 100
3 TABLE 1 Test compound Inhibition (%) Compound 109.2HCl (1 .mu.M)
81 (10 .mu.M) 100 Compound 165.2HCl.3/2H.sub.2O (10 .mu.M) 100
Compound 80.2HCl.H.sub.2O (10 .mu.M) 100 Compound 204.2HC1 (10
.mu.M) 93
Experimental Example 2
[0561] Rho Kinase Inhibitory Action (Inhibition of Human Platelet
Rho Kinase (p160ROCK))
[0562] Human platelet p160ROCK was isolated by the method of
Ishizaki et al. (Ishizaki T et al., The EMBO J., 15(8), 1885-1893,
1996).
[0563] Kinase assay included the following steps. That is, a
reaction mixture (total amount 30 .mu.l) containing 50 mM
Hepes-NaOH (pH =7.4), 10 mM MgCl.sub.2, 5 mM MnCl.sub.2, 2 mM
dithiothreitol, 0.02% Brij35, 1 .mu.M [.gamma.-.sup.32P]ATP, 330
.mu.g/ml histone, p160ROCK (2 .mu.l) isolated by the method of
Ishizaki et al. and the test compound was incubated at 30.degree.
C. for 20 minutes. The solution was mixed with a 1/3 amount of
4.times.Laemmli sample buffer, boiled for 5 minutes and applied to
SDS-PAGE. The gel was stained with Coomassie Brilliant Blue and
dried. The band of histone was cut out and assayed for
radioactivity. The test compound was evaluated in the same manner
as in Experimental Example 1, and the concentration of each test
compound necessary for 50% inhibition was calculated as IC50
(.mu.M). The results are shown in Table 2.
4 TABLE 2 Test compound IC.sub.50 (.mu.M) Compound 80.2HCl.H.sub.2O
1.5 Compound 109.2HCl 0.11 Compound 143.2HCl.H.sub.2O 1.6 Compound
204.2HCl 3.8 Compound 308.2HCl 5.0
Experimental Example 3
[0564] Rho Kinase Inhibitory Action (Inhibition of p160ROCK and
ROCKII)
[0565] The standard enzyme products of p160ROCK (Ishizaki T et al.,
The EMBO J., 15(8), 1885-1893, 1996) and ROCKII (Nakagawa 0 et al.,
FEBS Lett. 392 189-193, 1996) were obtained in the following
manner. COS cells were seeded in a 3.5 cm dish and incubated
overnight. Using lipofectamine, the expression vectors of p160ROCK
and ROCKII (pCAG-myc-p160ROCK and pCAG-myc-ROCKII: see Ishizaki T
et al., The EMBO J., 15(8), 1885-1893, 1996 and Nakagawa O et al.,
FEBS Lett. 392 189-193, 1996) were transfected. After incubation
for 20 hours, the cells were washed once with ice-cooled PBS, and
the cells were lysed on ice for 20 minutes using a lysis buffer (20
mM Tris-HCl (pH=7.5), 1 mM EDTA, 1 mM EGTA, 5 mM MgC.sub.2, 25 mM
NaF, 10 mM .beta. glycerophosphate, 5 mM sodium pyrophorphate, 0.2
mM phenylmethylsulfonyl fluoride, 2 mM dithiothreitol, 0.2 mM
sodium vanadate, 0.05% Triton X-100, 0.1 .mu.M calyculin A). The
lysate was centrifuged at 10,000.times.g for 10 minutes and the
supernatant was recovered. To the supernatant was added 9E10
anti-myc epitope antibody (see Ishizaki T et al., The EMBO J.,
15(8), 1885-1893, 1996) and the mixture was shaken for 2 hours.
Then, protein G-Sepharose was added and the mixture was shaken for
2 more hours. The suspension was centrifuged at 1,000.times.g for 5
minutes and the resulting pellets were washed 3 times with lysis
buffer and once with kinase buffer (50 mM Hepes-NaOH (pH=7.4), 10
mM MgC.sub.2, 5 mM MnCl.sub.2, 2 mM dithiothreitol, 0.02% Brij35).
The pellets were suspended in kinase buffer to give a standard
enzyme product. The kinase assay followed the method shown in
Experimental Example 2, wherein the standard enzyme product
obtained in this Experimental Example was used instead of human
platelet Rho kinase (p160ROCK). The concentration of each test
compound necessary for 50% inhibition was calculated as IC50
(.mu.M). The results are shown in Table 3.
5 TABLE 3 IC.sub.50 (.mu.M) Test compound p160ROCK ROCK-II Compound
80.2HCl.H.sub.2O 0.63 0.56 Compound 109.2HCl 0.095 0.048 Compound
143.2HCl.H.sub.2O 0.88 0.47 Compound 204.2HCl 2.3 1.1
[0566] Experimental Example 4
[0567] vasodilating action
[0568] Male rabbits (body weight 1.9-3.0 kg) were anesthetized with
pentobarbital sodium and exsanguinated, whereafter thoractic aorta
was removed. An about 2 mm width aortic ring samples were prepared
and hung in a Magnus bath (40 ml) filled with Krebs-Henseleit
solution (37.degree. C., NaCl 117 mM; KCl 4.7 mM; CaCl.sub.2 2.5
mM; MgSO.sub.4 1.2 mM; NaHCO.sub.3 24.8 mM; KH.sub.2PO.sub.4 1.2
mM; glucose 11.0 mM) at a load of 2 g. The Magnus bath was
constantly bubbled with a mixed gas (95% O.sub.2+5% CO.sub.2 gas).
The tension of the preparation was measured with an isomeric
transducer (TB-61 IT, Nippon Koden). The preparation was contracted
with phenylephrine (10.sup.-6 M) and, after the contraction was
stabilized, the test compound was added accumulatively and relaxing
action was observed. The relaxing action of the test compound was
calculated by expressing the concentration of the test compound
necessary for 50% relaxation as IC50 (.mu.M) against the
contraction with phenylephrine as 100%. The results are shown in
Table 4.
[0569] Experimental Example 5
[0570] Effect on contraction by acetylcholine of trachea specimen
removed from guinea pig
[0571] Male Hartley guinea pigs (body weight 260-390 g) were
anesthetized by the peritoneal administration of pentobarbital
sodium (100 mg/kg) and exsanguinated, whereafter trachea was
removed. The anterior cartilage of the trachea was opened and the
band was cut in a 3 mm width strip to give a specimen. The specimen
was hung in a Magnus bath (40 ml) filled with Krebs-Henseleit
solution (NaCl 117 mM; KCl 4.7 mM; CaCl.sub.2 2.5 mM; MgSO.sub.4
1.2 mM; NaHCO.sub.3 24.8 mM ; KH.sub.2PO.sub.4 1.2 mM ; glucose
11.0 mM) at a load of 1 g. The Magnus bath was constantly bubbled
with a mixed gas (95% O.sub.2+5% CO.sub.2 gas). The tension of the
strip was measured with an isomeric transducer (TB-61 1T, Nippon
Koden) and depicted on a recorder (Ti-102, Tokai Irika). The strip
was contracted with acetylcholine (10.sup.-6 M) and, after the
contraction was stabilized, the test compound was added
accumulatively and relaxing reaction was observed. The relaxing
action of the test compound was calculated and expressed by the
concentration of the test compound necessary for 50% relaxation as
IC50 (.mu.M) against the maximum response with papaverine
(10.sup.-4 M) as 100%. The results are shown in Table 4.
6 TABLE 4 Vasorelaxing Trachea relaxing Test compound action
(.mu.M) action (.mu.M) Compound 80.2HCl.H.sub.2O 0.70 0.56 Compound
109.2HCl 0.1 0.043 Compound 165.2HCl.{fraction (3/2)}H.sub.2O 0.051
0.066 Compound 179.2HBr.1/2H.sub.2O 0.03 0.029
Experimental Example 6
[0572] Peripheral Blood Flow Increasing Action
[0573] Streptozotocin (STZ, 65 mg/kg) was intravenously injected to
male SD rats (body weight 200-300 g) to prepare diabetic rats. One
month later, STZ-induced diabetic rats were anesthetized with
pentobarbital sodium and the blood flow in the hind limb skin was
measured with laser blood flowmeter (ALF21R, Advance). The test
compound was intravenously administered via catheter dwelled in the
carotid arteries, and hind limb skin blood flow increasing action
was observed. The blood flow increasing action of the test compound
was expressed by increase percentage from the blood flow before
administration. The results are shown in Table 5.
7TABLE 5 Increase in skin blood flow .+-. Test compound standard
error (%) Compound 80.2HCl.H.sub.2O (1 .mu.g) 135.0 .+-. 13.4
Compound 157.HCl.H.sub.2O (1 .mu.g) 211.6 .+-. 13.6 Compound
165.2HCl.{fraction (3/2)}H.sub.2O (0.03 .mu.g) 135.8 .+-. 0.0 (0.1
.mu.g) 144.7 .+-. 0.0 Compound 166.2HCl.H.sub.2O (0.3 .mu.g) 143.2
.+-. 25.4 (1 .mu.g) 165.9 .+-. 42.5
Experimental Example 7
[0574] Inhibition of VIA (Very Late Antigen) Integrin
Activation
[0575] As the index of the activation by VLA integrin, phorbol
ester-induced adhesion of CEM cells (human T cell type established
cell) to fibronectin, which is a ligand of VLA integrin, was
measured. The inhibitory action on the induced adhesion by the test
compound was determined by the following method.
[0576] CEM cells were washed with RPMI1640 medium containing 0.5%
bovine serum albumin (BSA), 10 mM HEPES, 2 mM L-glutamin, 1 mM
sodium pyruvate, 60 .mu.g/ml kanamycin sulfate and 1.5 mg/ml sodium
hydrogencarbonate (hereinafter this medium is referred to as
culture solution) and suspended in this medium for use in the
following experiment. To each well of a 96 well plate coated with
human fibronectin were added CEM cells (5.times.10.sup.4) and the
test compound dissolved in the culture solution (final
concentration 1-100 .mu.M) to the amount of 100 .mu.l, and the
plate was stood at 37.degree. C. for 1 hour. Then, PMA (phorbol
12-myristate 13-acetate, TPA; final concentration 10 ng/ml) and the
test compound were added to the amount of 200 .mu.l, and the plate
was stood at 37.degree. C. for 30 minutes. Each well was washed
twice with the culture solution (200 .mu.l) at 37.degree. C., and
the LDH (lactate dehydrogenase) activity of the cells adhered to
the plate was determined, whereby the amount of the adhered cell
was measured. Based on the results obtained by the above-mentioned
method, the inhibitory action of the test compound on the induced
adhesion was calculated by the following formula. The results are
shown in Table 6.
[0577] Inhibition (%) of adhesion
induction=(a-b)/(a-c).times.100
[0578] a=number of cells adhered with the addition of PMA
[0579] b=number of cells adhered with the addition of test compound
and PMA
[0580] c=number of cells adhered without stimulation
8TABLE 6 Adhesion induction Test compound Concentration (.mu.M)
inhibition (%) Compound 80.2HCl.H.sub.2O 100 70 Compound 109.2HCl
100 67 Compound 143.2HCl.H.sub.2O 100 77 Compound
165.2HCl.{fraction (3/2)}H.sub.2O 10 40 Compound 204.2HCl 100 82
Anti-.beta.1 antibody 20 .mu.g/ml 118 IgG1 20 .mu.g/ml -25
Experimental Example 8
[0581] Inhibition of Bone Resorption (In Vitro)
[0582] The determination of the in vitro inhibition of bone
resorption using mouse femoral bone followed the method below.
[0583] The femoral bone of 3-6 week old male ICR mice was
aseptically removed, and bone marrow cavity was washed with F12
medium, containing 10% heat inactivated fetal bovine serum,
penicillin G calcium (100 units/mil), kanamycin sulfate (60 .mu.g)
and 0.15% sodium hydrogencarbonate (hereinafter the medium is to be
referred to as culture solution). After washing the bone marrow
cavity and then removing the soft tissue adhered to the bone, the
bone was subjected to incubation. The test compound was once
dissolved in dimethyl sulfoxide (DMSO) to give a 10 mg/ml solution,
which was diluted 1000-fold with the culture solution to give a 10
.mu.g/ml solution. The test compounds were respectively added to
the concentration shown in Table 7 and, using this culture solution
(1.2 ml), the ICR mouse femoral bone was incubated in a 24 well
plate for 6 days under the conditions of 5% CO.sub.2 gas, 95% air.
After the completion of the incubation, the culture supernatant was
recovered, and the amount of calcium suspending in the culture
supernatant was quantitatively determined by the chelate method
using o-cresolphthalein. The bone resorption inhibitory action of
the test compound was calculated by the following formula using the
incubation of the femoral bone without addition of the test
compound as a control. 2 Inhibition of bone resorption ( % ) = (
Amount of free Ca without addition of test compound - Amount of
free Ca with addition of test compound Amount of free Ca without
addition of test compound - Amount of Ca in culture ) .times.
100
[0584] This experiment was done with 4 cases in each group. As the
control, the same amount of DMSO alone as in the case with the
addition of the test compound was used. The results are shown in
Table 7.
Experimental Example 9
[0585] Inhibition of Mouse Allogenic Mixed Lymphocyte Reaction
[0586] A mouse allogenic mixed lymphocyte reaction (hereinafter to
be referred to as mouse allogenic MLR) was performed by mixed
culture (equal ratio) of the spleen cell of BALB/c mice as the
reaction cell and the spleen cell of C57BL/6 mice treated with
mitomycin C as stimulated cell.
[0587] The reaction cells were prepared by the following method.
Spleen was removed from 5-6 week old BALB/c mice and treated with
RPMI1640 medium (containing kanamycin sulfate (60 .mu.g/ml),
penicillin G potassium (100 units/ml),
N-2-hydroxyethylpiperazine-N'-2-ethanesulfonate (10 mM), 0.1%
sodium hydrogencarbonate and L-glutamin (2 mM)) supplemented with
5% heat inactivated fetal bovine serum (FBS) to give a single cell
suspension of the spleen cell. After hemolysis treatment, the
suspension was adjusted to 10.sup.7 cells/ml with RPMI 1640 medium
containing 10.sup.-4M 2-mercaptoethanol and 10% FBS and used as a
reaction cell suspension.
[0588] The reaction cell suspension (50 .mu.l) prepared by the
above method, stimulated cell suspension (50 .mu.l) and the test
compound (100 .mu.l) prepared using RPMI1640 medium containing 10%
FBS were added to a 96 well plate and incubated at 37.degree. C.
under 5% CO.sub.2 gas, 95% air for 4 days.
[0589] A pigment assay using
3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetr- azolium bromide
(MTT) was applied for the determination of lymphocyte
transformation reaction.
[0590] After the completion of culture, the supernatant (100 .mu.l)
in each well was removed, and 5 mg/ml MTT solution (20 .mu.l) was
added to each well, which was followed by incubation at 37.degree.
C. for 4 hours. Then, a 0.01 N hydrochloric acid solution (100
.mu.l) containing 10% sodium dodecyl sulfate was added and the
mixture was incubated at 37.degree. C. overnight. The resulting
purple crystals of formazan was dissolved and absorbance at 550 nm
was measured using a microplate absorption meter, which was used as
the index of lymphocyte transformation reaction of the mouse
allogenic MLR. The inhibition of mouse allogenic MLR was evaluated
by calculating the inhibition percentage by the following formula.
The results are shown in Table 7.
9TABLE 7 3 Inhibition ( % ) = [ 1 - Absorbance of MLR with addition
of test compound - absorbance of reacted cells alone Absorbance of
MLR without addition of test compound - absorbance of reacted cells
alone ] .times. 100 Mouse allogenic MLR Bone resorption inhibitory
activity Test compound inhibition % (.mu.M) IC.sub.50 (.mu.M)
Compound 80.2HCl.H.sub.2O 40.9(100) 9.6 Compound 109.2HCl 42.6(100)
1.6 Compound 112.2HCl 75.7(100) 4.4 Compound 110.2HCl.H.sub.2O
74.0(100) 1.1 Compound 142.2HCl.H.sub.2O 44.2(100) Compound
143.2HCl.H.sub.2O 39.4(100) Compound 308.2HCl 13.9
Experimental Example 10
[0591] Inhibition of Cell Growth of SK-Mel-28 Melanoma
[0592] Human SK-Mel-28 melanoma (10.sup.4 cells) and the test
compound were suspended in RPMI1640 medium containing 100 .mu.l of
10% FBS and incubated in a 96 well plate at 37.degree. C. under 5%
CO.sub.2 gas for 72 hours. After the incubation, 10 .mu.l of MTT (5
mg/ml) was added to each well and the cells were incubated at
37.degree. C. under 5% CO.sub.2 gas for 4 hours. Then, 10% sodium
dodecyl sulfate and 0.01 N hydrochloric acid solution were each
added by 10 .mu.l to respective wells. After the plate was stood
overnight, absorbance at 570 nm was measured using a microplate
reader and the inhibition percentage (% cytotoxicity) was
calculated by the following formula. The results are shown in Table
8.
[0593] The cytotoxicity against human cultured tumor cells was
confirmed by pigment method (Carmichael et al., Cancer Res., 47,
936-942, 1987 : Mosman, J. Immunol. Methods, 65, 55-63, 1983) using
3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide
(MTT).
[0594] The test compound was dissolved in dimethyl sulfoxide and
diluted with RPMI1640 medium before use. The final dimethyl
sulfoxide concentration was adjusted to not more than 0.25%. 4
Inhibition ( % ) = [ 1 - Absorbance when test compound was added
Absorbance when test compound was not added ] .times. 100
10TABLE 8 Test compound Cell growth inhibition IC.sub.50 (.mu.M)
Compound 115.2HBr.1/4H.sub.2O 9 Compound 109.2HCl 58 Compound
142.2HCl.H.sub.2O 59 Compound 145.2HCl.H.sub.2O 62
Experimental Example 11
[0595] Inhibition of Angiogenesis
[0596] The inhibition of angiogenesis was evaluated by using the
inhibition of lumen formation in vascular endothelial cell as an
index. To be specific, normal human umberical vascular endothelial
cells (KURABO INDUSTRIES LTD.) were suspended in E-GMUV medium at
5.5.times.10.sup.4 cells/ml and 400 .mu.l therefrom was added on
matrigel plate (EHS sarcoma-derived reconstructed basement
membrane, Collaborative Biomedical Products). Then, the test
compound (1 mM solution, 4 .mu.l) was added and the cells were
incubated at 37.degree. C. under 5% CO.sub.2 gas for 18 hours.
After the completion of the incubation, the number of lumen per
predetermined area was counted under a microscope. Inasmuch as the
number of lumen increases by the inhibition of lumen formation, the
test compound was evaluated by comparison of the number of lumen
with the control. The results are shown in Table 9.
11 TABLE 9 Test compound Number of lumen (10 .mu.M) Compound
109.2HCl 153% Compound 80.2HCl.H.sub.2O 174% Compound
110.2HCl.H.sub.2O 203% Compound 165.2HCl.3/2H.sub.2O 222% Compound
204.2HCl 133%
Experimental Example 12
[0597] Inhibition of Growth of Vascular Smooth Muscle Cell
[0598] The separation from the artery of rat and culture of smooth
muscle cell (SMC) followed the explant method of Ross (Ross, R. and
Glomset, J. A., N. Engl. J. Med., 295, 369-420, 1976). Male wistar
rats (10 week old) was slaughtered by cutting the carotid arteries
and aorta of thorax was removed. After removal of fat tissues
around the tunica externa and peeling of tunica intima, the artery
was minced and incubated in 10% fetal bovine serum (FBS)-containing
DMEM medium at 37.degree. C. under 5% CO.sub.2 gas. Seven days
later, the out-grown cells were separated by trypsin treatment,
washed with phosphate-buffered saline (PBS) and incubated in 10%
FBS-containing DMEM medium in a 80 cm.sup.2 culture flask. The
cells of subculture 2 were suspended in 10% FBS-containing DMEM
medium at 5.times.10.sup.4 cells/ml and 100 .mu.l thereof per well
was added to 96 well collagen-coated plate, which was incubated at
37.degree. C. under 5% CO.sub.2 gas for one day. The test compound
was appropriately diluted with dimethyl sulfoxide (DMSO) and added
to the 96 well plate. The concentration of DMSO in the medium was
adjusted to 1%. After 48 hours, 10 .mu.l of MTT solution (5 mg/ml)
was added and, 4 hours later, 10% sodium dodecyl sulfate-0.01 N
hydrochloric acid (50 .mu.l) was added. The absorbance at 570 nm
was measured the following day by an immunoreader. The SMC growth
inhibitory action of the test compound was shown by inhibition
percentage calculated by the following formula. The results are
shown in Table 10. 5 Inhibition ( % ) = [ 1 - Absorbance when test
compound was added Absorbance when test compound was not added ]
.times. 100
12 TABLE 10 Test compound IC.sub.50 (.mu.M) compound 153.2HCl 27
compound 157.2HCl.H.sub.2O 55 compound 165.2HCl.3/2H.sub.2O 38
compound 163.2HBr 63
Experimental Example 13
[0599] Acute Toxicity
[0600] The compound 109.2HCl and compound 143.2HCl.H.sub.2O were
respectively administered intraperitoneally to ddY mice and the
mice were monitored for 5 days. As a result, the intraperitoneal
administration at 30 mg/kg did not cause death.
[0601] The foregoing Formulation Examples and pharmacological
experiments reveal that the compounds of the formula (I) and the
formula (1) have strong Rho kinase inhibitory action. These Rho
kinase inhibitors have vasodilating action, trachea relaxing
action, peripheral blood flow increasing action, cell adhesion
induction inhibitory action, tumor cell metastasis inhibitory
action, bone resorption inhibitory action, mouse allogenic MLR
inhibitory activity, tumor cell growth inhibitory action,
angiogenesis inhibitory action, vascular smooth muscle cell growth
inhibitory action and other various actions. Therefore, they are
useful as pharmaceutical agents, particularly, a therapeutic agent
of hypertension, a therapeutic agent of angina pectoris, a
suppressive agent of cerebrovascular contraction, a therapeutic
agent of asthma, a therapeutic agent of peripheral circulation
disorder, a prophylactic agent of immature birth, a therapeutic
agent of arteriosclerosis, an anti-cancer drug, an
anti-inflammatory agent, an immunosuppressant, a therapeutic agent
of autoimmune disease, an anti-AIDS drug, a contraceptive, a
prophylactic agent of digestive tract infection, a therapeutic
agent of osteoporosis, a therapeutic agent of retinopathy and a
brain function improving drug.
[0602] In addition, since Rho kinase inhibitors of the present
invention have strong Rho kinase inhibitory activity, they are also
useful as reagents for the study relating to Rho and Rho kinase and
as diagnostics of the diseases related to them.
[0603] This application is based on application No. 212409/1996
filed in Japan, the contents of which are incorporated hereinto by
reference.
* * * * *