U.S. patent application number 10/702427 was filed with the patent office on 2004-05-13 for immunosuppressive imidazole derivatives and their combination preparations with tacrolimus of cyclosporins.
This patent application is currently assigned to Fujisawa Pharmaceutical Company, Ltd.. Invention is credited to Chida, Noboru, Nakayama, Osamu, Sakai, Fumihiko, Yamazaki, Harumi, Yokota, Yoshihiro.
Application Number | 20040092436 10/702427 |
Document ID | / |
Family ID | 27158071 |
Filed Date | 2004-05-13 |
United States Patent
Application |
20040092436 |
Kind Code |
A1 |
Sakai, Fumihiko ; et
al. |
May 13, 2004 |
Immunosuppressive imidazole derivatives and their combination
preparations with tacrolimus of cyclosporins
Abstract
Present invention is relating to a new use of a compound
possessing an inhibitory activity on the production of nitric
oxide, for increasing an effect caused by IL-2 inhibitor.
Inventors: |
Sakai, Fumihiko;
(Tsukuba-shi, JP) ; Yamazaki, Harumi;
(Niihari-gun, JP) ; Chida, Noboru;
(Nishinomiya-shi, JP) ; Nakayama, Osamu;
(Kitasouma-gun, JP) ; Yokota, Yoshihiro;
(Nishinomiya-shi, JP) |
Correspondence
Address: |
LEYDIG VOIT & MAYER, LTD
700 THIRTEENTH ST. NW
SUITE 300
WASHINGTON
DC
20005-3960
US
|
Assignee: |
Fujisawa Pharmaceutical Company,
Ltd.
Yodogawa-ku
JP
532-8514
|
Family ID: |
27158071 |
Appl. No.: |
10/702427 |
Filed: |
November 7, 2003 |
Related U.S. Patent Documents
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Application
Number |
Filing Date |
Patent Number |
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10702427 |
Nov 7, 2003 |
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09646886 |
Dec 1, 2000 |
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6673807 |
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09646886 |
Dec 1, 2000 |
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PCT/JP99/01806 |
Apr 5, 1999 |
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Current U.S.
Class: |
424/278.1 ;
514/20.5; 514/291 |
Current CPC
Class: |
A61P 37/06 20180101;
A61K 31/4178 20130101; A61K 31/00 20130101; A61P 43/00 20180101;
A61K 31/4439 20130101 |
Class at
Publication: |
514/011 ;
514/291 |
International
Class: |
A61K 038/13; A61K
031/4745 |
Foreign Application Data
Date |
Code |
Application Number |
Apr 6, 1998 |
AU |
PP2827 |
May 4, 1998 |
AU |
PP3324 |
Aug 4, 1998 |
AU |
PP5056 |
Claims
What is claimed is:
1. A method for increasing graft survival time in a transplantation
subject having a graft comprising: administering to said subject
compound (a) possessing an inhibitory activity on the production of
nitric oxide and compound (b) that is a cyclosporin or a rapamycin,
in an amount and for a time and under conditions effective to
increase graft survival time.
2. The method of claim 1 comprising administering a
cyclosporin.
3. The method of claim 1 comprising administering a rapamycin.
Description
CROSS-REFERENCE TO RELATED PATENT APPLICATIONS
[0001] This patent application is a Divisional of copending U.S.
patent application Ser. No. 09/646,886, filed Dec. 1, 2000,
pending, which is a 35 USC 371 of application PCT/JP99/01806, filed
Apr. 5, 1999.
TECHNICAL FIELD
[0002] This invention relates to a new use of a compound possessing
an inhibitory activity on the production of nitric oxide, which is
useful in a medical field.
BACKGROUND ART
[0003] Various compounds possessing an inhibitory activity on the
production of nitric oxide have already been known, for example, in
EP 0 394 989-A2, WO96/16981, WO97/45425, WO98/27108, etc.
DISCLOSURE OF INVENTION
[0004] This invention relates to a new use of a compound possessing
an inhibitory activity on the production of nitric oxide, for
increasing an effect caused by interleukin 2 inhibitor
(hereinafter, referred to IL-2 inhibitor).
[0005] Therefore, one object of the present invention is to provide
a new use of a compound possessing an inhibitory activity on the
production of nitric oxide, for increasing an effect caused by IL-2
inhibitor.
[0006] Another object of this invention is to provide a method for
increasing an effect caused by IL-2 inhibitor by administering an
effective amount of a compound possessing an inhibitory activity on
the production of nitric oxide.
[0007] A further object of this invention is to provide a use of a
compound possessing an inhibitory activity on the production of
nitric oxide for manufacturing a medicament for increasing an
effect caused by IL-2 inhibitor.
[0008] Still further object of this invention is to provide a
composition comprising a compound possessing an inhibitory activity
on the production of nitric oxide, for increasing an effect caused
by IL-2 inhibitor.
[0009] In the present invention, the "compound possessing an
inhibitory activity on the production of nitric oxide" should not
be limited and be considered to mean any compounds which have an
inhibitory activity on the production of nitric oxide. Preferable
one is a compound possessing an inhibitory activity on the
production of inducible nitric oxide synthase (iNOS), and the other
preferable one is a compound possessing an iNOS-inhibitory
activity.
[0010] The compound having the following formula (I) are
exemplified as a preferable example of the above "compound
possessing an inhibitory activity on the production of nitric
oxide". 1
[0011] wherein
[0012] R.sup.1 is indolyl which may have a suitable substituent
selected from the group consisting of lower alkyl, phenyl, halogen,
lower alkoxy, and nitro, benzofuranyl, phenyl which may have one or
two suitable substituent(s) selected from the group consisting of
amino, acylamino, lower alkylamino, halogen, lower alkoxy and
nitro, lower alkyl, quinoxalinyl, quinolyl, pyrrolyl, pyrimidinyl
having benzofuranyl, benzimidazolyl, benzothienyl, benzothiazolyl,
benzoxazolyl, indolinyl, anilino, phenylcarbamoyl or imidazolyl
which may have one or two suitable substituent(s) selected from the
group consisting of phenyl, lower alkyl and indolyl;
[0013] R.sup.2 is hydrogen or phenyl(lower)alkyl;
[0014] R.sup.4 is hydrogen, phenyl or pyridyl, each of which may
have suitable substituent(s) selected from the group consisting of
lower alkyl, lower alkoxy, lower alkylthio, halogen, trihalomethyl,
nitro, cyano, imidazolyl, optionally protected hydroxy, acyl,
amino, acylamino, diacylamino, di(lower)alkyl-amino,
amino(lower)alkyl, acylamino(lower)alkyl, pyrazolyl, morpholinyl,
piperidyl, triazolyl, lower alkoxy(lower)alkoxy,
hydroxy(lower)alkyl, lower alkylpiperazinyl, phenyl and carboxy,
quinolyl or 3,4-methylenedioxyphenyl;
[0015] R.sup.5 is hydrogen, imidazolyl, phenyl, nitrophenyl,
phenyl(lower)-alkyl, optionally esterified carboxy or a group of
the formula: 2
[0016] in which R.sup.7 and R.sup.8 are the same or different and
each is hydrogen, phenyl, phenyl(lower)alkyl, lower alkyl or lower
alkoxy; or R.sup.4 and R.sup.5 in combination form a group of the
formula:
--CH.dbd.CH--CH.dbd.CH--
[0017] Y is a group of the formula: 3
[0018] in which R.sup.3 is hydrogen or a group of the formula:
--(CH2)n-R.sup.6
[0019] in which R.sup.6 is optionally protected hydroxy, acyl,
carboxy, acylamino, lower alkoxy, phenyl(lower)alkoxy, lower
alkylthio, phenyl which may have a suitable substituent selected
from the group consisting of lower alkoxy, halogen, amino,
acylamino, diacylamino and nitro, pyridyl which may have a suitable
substituent selected from the group consisting of lower alkoxy and
halogen, pyrazinyl, pyrimidinyl, furyl, imidazolyl, naphthyl,
N-(lower)alkylindolyl or 3,4-methylenedioxyphenyl, and n is an
integer of 0 to 3, or a group of the formula: 4
[0020] in which R.sup.11 is phenyl, phenoxy or phenyl(lower)alkoxy;
or R.sup.2 and R.sup.3 in combination form a group of the formula:
5
[0021] m is 0 or 1; and
[0022] X is S or NR.sup.9
[0023] in which R.sup.9 is hydrogen, lower alkyl, cyclo(lower)alkyl
or a group of the formula: 6
[0024] in which R.sup.10 is hydrogen, lower alkyl or lower alkoxy;
or a pharmaceutically acceptable salt thereof.
[0025] Suitable pharmaceutically acceptable salts of the compound
(I) are conventional non-toxic salts and include, for example, a
salt with a base or an acid addition salt such as a salt with an
inorganic base, for example, an alkali metal salt (e.g., sodium
salt, potassium salt, etc.), an alkaline earth metal salt (e.g.,
calcium salt, magnesium salt, etc.), an ammonium salt; a salt with
an organic base, for example, an organic amine salt (e.g.,
triethylamine salt, pyridine salt, picoline salt, ethanolamine
salt, triethanolamine salt, dicyclohexylamine salt,
N,N'-dibenzylethylenediamine salt, etc.); an inorganic acid
addition salt (e.g., hydrochloride, hydrobromide, sulfate,
phosphate, etc.); an organic carboxylic or sulfonic acid addition
salt (e.g., formate, acetate, trifluoroacetate, maleate, tartrate,
citrate, fumarate, methanesulfonate, benzenesulfonate,
toluenesulfonate, etc.); and a salt with a basic or acidic amino
acid (e.g., arginine, aspartic acid, gultamic acid, etc.).
[0026] In the above and subsequent descriptions of the present
specification, suitable examples and illustration of the various
definitions which the present invention intends to include within
the scope thereof are explained in detail as follows.
[0027] The term "lower" is used to intend a group having 1 to 6,
preferably 1 to 4, carbon atom(s), unless otherwise provided.
[0028] Suitable "lower alkyl" and "lower alkyl moiety" in the terms
"lower alkylthio", "lower alkylthio(lower)alkyl",
"N-(lower)alkylindolyl", "lower alkylamino", di(lower)alkylamino",
"phenyl(lower)alkyl", "amino(lower)alkyl",
"acylamino-(lower)alkyl", "hydroxy(lower)alkyl" and "lower
alkyl-piperazinyl" include straight or branched one having 1 to 6
carbon atom(s), such as methyl, ethyl, propyl, isopropyl, butyl,
isobutyl, sec-butyl, tert-butyl, pentyl, tert-pentyl and hexyl, in
which more preferred one is C1-C4 alkyl.
[0029] Suitable "lower alkoxy" and "lower alkoxy moiety" in the
terms "lower alkoxy(lower)alkoxy" and "phenyl(lower)alkoxy"
include, for example, methoxy, ethoxy, propoxy, isopropoxy, butoxy,
isobutoxy, tert-butoxy, pentyloxy, tert-pentyloxy and hexyloxy, in
which more preferred one is C1-C4 alkoxy.
[0030] Suitable "halogen" includes, for example, fluorine, bromine,
chlorine and iodine.
[0031] "Optionally esterified carboxy" includes carboxy and
esterified carboxy. Suitable examples of said ester include lower
alkyl ester (e.g., methyl ester, ethyl ester, propyl ester,
isopropyl ester, butyl ester, isobutyl ester, tert-butyl ester,
pentyl ester, tert-pentyl ester, hexyl ester, etc.); lower alkenyl
ester (e.g., vinyl ester, allylester, etc.); lower alkynyl ester
(e.g., ethynyl ester, propynyl ester, etc.); lower
alkoxy(lower)alkyl ester (e.g., methoxymethyl ester, ethoxymethyl
ester, isopropoxymethyl ester, 1-methoxyethyl ester, 1-ethoxyethyl
ester, etc.); mono(or di or tri)-aryl(lower)alkyl ester, for
example, mono(or di or tri)phenyl-(lower)alkyl ester which may have
one or more suitable substituent(s) [e.g., benzyl ester,
4-methoxybenzyl ester, 4-nitrobenzyl ester, phenethyl ester, trityl
ester, benzhydryl ester, bis(methoxyphenyl)methyl ester,
3,4-dimethoxybenzyl ester, 4-hydroxy-3,5-di-tert-butylbenzyl ester,
etc.]; and aryl ester which may have one or more suitable
substituent(s) such as substituted or unsubstituted phenyl ester
(e.g., phenyl ester, tolyl ester, tert-butylphenyl ester, xylyl
ester, mesityl ester, cumenyl ester, 4-chlorophenyl ester,
4-methoxyphenyl ester, etc.).
[0032] Suitable "trihalomethyl" includes, for example,
trifluoromethyl, trichloromethyl and tribromomethyl, in which
preferred one is trifluoromethyl.
[0033] Suitable "amino protective group" includes, for example,
acyl and conventional protective group such as mono(or di or
tri)aryl(lower)-alkyl, for example, mono(or di or
tri)phenyl(lower)alkyl (e.g., benzyl, trityl, etc.).
[0034] Suitable "acyl" and "acyl moiety" in the terms "acylamino",
"diacylamino" and "acylamino(lower)alkyl" include, for example,
carbamoyl which may be substituted by suitable substituent(s),
aliphatic acyl group and acyl group containing an aromatic ring,
which is referred to as aromatic acyl, or a heterocyclic ring,
which is referred to as heterocyclic acyl.
[0035] Suitable examples of said acyl are illustrated as follows:
"carbamoyl which may be substituted by suitable substituent(s)"
includes a group of the formula: 7
[0036] wherein R.sup.12 and R.sup.13 are the same or different and
each is hydrogen, lower alkyl, phenyl which may have a suitable
substituent selected from the group consisting of lower alkoxy and
halogen, phenyl(lower)alkyl, pyridyl, pyridyl(lower)alkyl or
3,4-ethylenedioxyphenyl; aliphatic acyl such as lower alkanoyl
which may be substituted by one to three halogen atoms (e.g.,
formyl, acetyl, propanoyl, butanoyl, 2-methylpropanoyl, pentanoyl,
2,2-dimethylpropanoyl, hexanoyl, trichloroacetyl, trifluoroacetyl,
etc.), lower alkoxycarbonyl (e.g., methoxycarbonyl, ethoxycarbonyl,
tert-butoxycarbonyl, tert-pentyloxycarbonyl, etc.), lower
alkyl-sulfonyl (e.g., methylsulfonyl, ethylsulfonyl, etc.), lower
alkoxysulfonyl (e.g., methoxysulfonyl, ethoxysulfonyl, etc.),
cyclo(lower)alkylcarbonyl (e.g., cyclopentylcarbonyl,
cyclo-hexylcarbonyl, etc.), and the like; aromatic acyl such as
aroyl (e.g., benzoyl, toluoyl, naphthoyl, etc.),
aryl(lower)alkanoyl [e.g., phenyl(lower)alkanoyl (e.g.,
phenylacetyl, phenylpropanoyl, phenylbutanoyl, etc.),
naphthyl(lower)alkanoyl (e.g., naphthylacetyl, naphthyl-propanoyl,
naphthylbutanoyl, etc.), etc.], aryl(lower)alkoxy-carbonyl [e.g.,
henyl(lower)alkoxycarbonyl (e.g., benzyloxy-carbonyl, etc.), etc.],
aryloxycarbonyl (e.g., phenoxycarbonyl, naphthyloxycarbonyl, etc.),
aryloxy(lower)alkanoyl (e.g., phenoxyacetyl, phenoxypropionyl,
etc.), arylsulfonyl (e.g., phenylsulfonyl, p-tolylsufonyl, etc.),
and the like; heterocyclic acyl such as indolylcarbonyl (e.g.,
indolyl-2-yl-carbonyl, etc.), benzofuranylcarbonyl (e.g.,
benzofuran-2-yl-carbonyl), quinoxalinylcarbonyl, quinolylcarbonyl,
pyrrolyl-carbonyl, benzimidazolylcarbonyl, benzothienylcarbonyl,
benzothiazolylcarbonyl, imidazolylcarbonyl, pyridylcarbonyl,
morpholinylcarbonyl (e.g., morpholinocarbonyl) and the like.
"Optionally protected hydroxy" includes hydroxy and protected
hydroxy. Suitable examples of "hydroxy protective group" in the
term "protected hydroxy" include acyl (e.g., acetyl,
trichloroacetyl, etc.), mono(or di or tri)phenyl(lower)alkyl which
may have one or more suitable substituent(s) (e.g., benzyl,
4-methoxybenzyl, trityl, etc.), trisubstituted silyl [e.g.,
tri(lower)alkylsilyl (e.g., trimethylsilyl,
tert-butyl-dimethylsilyl, etc.), etc.], tetrahydropyranyl and the
like.
[0037] Suitable "protected carboxy" is carboxy group protected by
conventional protective group such as lower alkoxycarbonyl [e.g.,
methoxycarbonyl, ethoxycarbonyl, propoxycarbonyl,
isopropoxycarbonyl, butoxycarbonyl, sec-butoxycarbonyl,
isobutoxycarbonyl, tert-butoxycarbonyl, pentyloxycarbonyl,
neopentyloxycarbonyl, hexyloxycarbonyl, etc.], optionally
substituted phenyl(lower)alkoxycarbon- yl for example, mono- or di-
or tri-phenyl(lower)alkoxycarbonyl which may be substituted by
nitro[e.g., benzyloxycarbonyl, 4-nitrobenzyloxycarbonyl- ,
benzhydryloxycarbonyl, trityloxycarbonyl, etc.] and the like.
[0038] Suitable "cyclo(lower)alkyl" includes cycloalkyl having 3 to
6 carbon atoms, such as cyclopropyl, cyclobutyl, cyclopentyl and
cyclohexyl, in which more preferred ones are cyclopropyl and
cyclobutyl.
[0039] The term "morpholinyl" includes 2-morpholinyl, 3-morpholinyl
and 4-orpholinyl (i.e. morpholino).
[0040] The term "piperidyl" includes 1-piperidyl (i.e. piperidino),
2-piperidyl, 3-piperidyl and 4-piperidyl.
[0041] And further, the compound having the following formula (II)
is also exemplified as the preferable one of the "compound
possessing an inhibitory activity on the production of nitric
oxide". 8
[0042] wherein,
[0043] R.sup.21 is benzofuranyl having halogen,
[0044] R.sup.22 is lower alkyl, and
[0045] R.sup.23 is morpholinyl.
[0046] The compound (I) and (II) usable in the present invention
can be prepared in a similar manner to that of WO98/27108
(PCT/JP97/04243), the disclosure of which is incorporated herein by
reference.
[0047] It is to be noted that the compound (I), (II) and other
compounds may include one or more stereoisomer(s) such as optical
isomer(s) and geometrical isomer(s) due to asymmetric carbon
atom(s) and double bond(s), and all of such isomers and mixtures
thereof are included within the scope of this invention.
[0048] The compound (I), (II) and pharmaceutically acceptable salts
thereof can be in a form of solvates [e.g., hydrate, ethanolate,
etc.], which are included within the scope of the present
invention.
[0049] And further, the compound (I), (II) and pharmaceutically
acceptable salts thereof can be in a form of pro-drugs, suitable
derivatives, and so on.
[0050] The compound (I), (II) and pharmaceutically acceptable salts
thereof possesses a strong inhibitory activity on the production of
nitric oxide (NO). They are expected to possess a nitric oxide
synthase (NOS)-inhibitory activity or a NOS-production inhibitory
activity.
[0051] And further, the compounds shown in EP 0 394 989-A2, WO
96/16981, WO 97/45425, the Japanese Patent publication No.
10-45751, the Japanese patent application No. 9-160128, and so on,
can also be used as "the compound possessing the inhibitory
activity on the production of nitric oxide".
[0052] The "IL-2 inhibitor" used in the present invention should
not be limited and be considered to mean any ones possessing IL-2
inhibitory activity. The particular example is the one possessing
an inhibitory activity on the production of IL-2. And the other is
the one that inhibits the transmission of IL-2 signal.
[0053] IL-2 is known to mediate immune system. Therefore, the
preferable "effect caused by interleukin 2 inhibitor" is an
immunosuppressive activity. Particularly, "the effect caused by
interleukin 2 inhibitor" may be the treatment and prevention of
rejection by transplantation, Graft-versus-Host diseases by medulla
ossium transplantation, autoimmune diseases, and so on. And the
present invention is useful to suppress immune reaction, to prolong
the survival period of the graft, to reduce the administration
amount of IL-2 inhibitor, and/or to reduce undesirable side effects
caused by IL-2 inhibitor.
[0054] Preferable "IL-2 inhibitor" is, for example, the tricyclic
macrolide shown in EP-0184162, WO89/05303, WO93/05058, WO96/31514,
and so on, the disclosure of which is incorporated herein by
reference. It is well known that those tricyclic macrolides have
strong IL-2 inhibitory activity.
[0055] As a particular example of the tricyclic macrolides
compounds, the tricyclic compound of the following formula (III)
can be exemplified. 9
[0056] (wherein each of adjacent pairs of R.sup.1 and R.sup.2,
R.sup.3 and R.sup.4, and R.sup.5 and R.sup.6 independently
[0057] (a) is two adjacent hydrogen atoms, but R.sup.2 may also be
an alkyl group or
[0058] (b) may form another bond formed between the carbon atoms to
which they are attached;
[0059] R.sup.7 is a hydrogen atom, a hydroxy group, a protected
hydroxy group, or an alkoxy group, or an oxo group together with
R.sup.1;
[0060] R.sup.8 and R.sup.9 are independently a hydrogen atom or a
hydroxy group;
[0061] R.sup.10 is a hydrogen atom, an alkyl group, an alkyl group
substituted by one or more hydroxy groups, an alkenyl group, an
alkenyl group substituted by one or more hydroxy groups, or an
alkyl group substituted by an oxo group;
[0062] X is an oxo group, (a hydrogen atom and a hydroxy group), (a
hydrogen atom and a hydrogen atom), or a group represented by the
formula --CH.sub.2O--;
[0063] Y is an oxo group, (a hydrogen atom and a hydroxy group), (a
hydrogen atom and a hydrogen atom), or a group represented by the
formula N--NR.sup.11R.sup.12 or N--OR.sup.13;
[0064] R.sup.11 and R.sup.12 are independently a hydrogen atom, an
alkyl group, an aryl group or a tosyl group;
[0065] R.sup.13, R.sup.14, R.sup.15, R.sup.16, R.sup.17, R.sup.18,
R.sup.19, R.sup.22 and R.sup.23 are independently a hydrogen atom
or an alkyl group;
[0066] R.sup.24 is an optionally substituted ring system which may
contain one or more heteroatoms;
[0067] n is an integer of 1 or 2; and
[0068] in addition to the above definitions, Y, R.sup.10 and
R.sup.23, together with the carbon atoms to which they are
attached, may represent a saturated or unsaturated 5- or 6-membered
nitrogen, sulfur and/or oxygen containing heterocyclic ring
optionally substituted by one or more groups selected from the
group consisting of an alkyl, a hydroxy, an alkoxy, a benzyl, a
group of the formula --CH.sub.2Se(C.sub.6H.sub.5), and an alkyl
substituted by one or more hydroxy groups.
[0069] Preferable R.sup.24 may be cyclo(C.sub.5-7)alkyl group, and
the following ones can be exemplified.
[0070] (a) a 3,4-di-oxo-cyclohexyl group;
[0071] (b) a 3-R.sup.20-4-R.sup.21-cyclohexyl group, in which
R.sup.20 is hydroxy, an alkoxy group, an oxo group, or a
--OCH.sub.2OCH.sub.2CH.sub.2- OCH.sub.3 group, and
[0072] R.sup.21 is hydroxy, --OCN, an alkoxy group, a heteroaryloxy
which may be substituted by suitable substituents, a
--OCH.sub.2OCH.sub.2CH.sub- .2OCH.sub.3 group, a protected hydroxy
group, chloro, bromo, iodo, aminooxalyloxy, an azido group,
p-tolyloxythiocarbonyloxy,
[0073] or R.sup.25R.sup.26CHCOO--, in which R.sup.25 is optionally
protected hydroxy or protected amino, and
[0074] R.sup.26 is hydrogen or methyl, or
[0075] R.sup.20 and R.sup.21 together form an oxygen atom in an
epoxide ring; or
[0076] (c) cyclopentyl group substituted by methoxymethyl,
optionally protected hydroxymethyl, acyloxymethyl (in which the
acyl moiety optionally contains either a methylamino group which
may be quaternized, or a carboxy group which may be esterified),
one or more amino and/or hydroxy groups which may be protected, or
aminooxalyloxymethyl. A preferred example is a 2-formyl-cyclopentyl
group.
[0077] The definitions used in the above general formula (III) and
the specific and preferred examples thereof are now explained and
set forth in detail.
[0078] The term "lower" means, unless otherwise indicated, a group
having 1 to 6 carbon atoms.
[0079] Preferable examples of the "alkyl groups" and an alkyl
moiety of the "alkoxy group" include a straight or branched chain
aliphatic hydrocarbon residue, for example, a lower alkyl group
such as methyl, ethyl, propyl, isopropyl, butyl, isobutyl, pentyl,
neopentyl and hexyl.
[0080] Preferable examples of the "alkenyl groups" include a
straight or branched chain aliphatic hydrocarbon residue having one
double-bond, for example, a lower alkenyl group such as vinyl,
propenyl (e.g., allyl group), butenyl, methylpropenyl, pentenyl and
hexenyl.
[0081] Preferable examples of the "aryl groups" include phenyl,
tolyl, xylyl, cumenyl, mesityl and naphthyl.
[0082] Preferable protective groups in the "protected hydroxy
groups" and the "protected amino" are 1-(lower
alkylthio)-(lower)alkyl group such as a lower alkylthiomethyl group
(e.g., methylthiomethyl, ethylthiomethyl, propylthiomethyl,
isopropylthiomethyl, butylthiomethyl, isobutylthiomethyl,
hexylthiomethyl, etc.), more preferably C.sub.1-C.sub.4
alkylthiomethyl group, most preferably methylthiomethyl group;
trisubstituted silyl group such as a tri(lower)alkylsilyl (e.g.,
trimethylsilyl, triethylsilyl, tributylsilyl,
tert-butyldimethylsilyl, tri-tert-butylsilyl, etc.) or lower
alkyl-diarylsilyl (e.g., methyldiphenylsilyl, ethyldiphenylsilyl,
propyldiphenylsilyl, tert-butyldiphenylsilyl, etc.), more
preferably tri(C.sub.1-C.sub.4)alkyl- silyl group and
C.sub.1-C.sub.4 alkyldiphenylsilyl group, most preferably
tert-butyldimethylsilyl group and tert-butyldiphenylsilyl group;
and an acyl group such as an aliphatic, aromatic acyl group or an
aliphatic acyl group substituted by an aromatic group, which are
derived from a carboxylic acid, sulfonic acid or carbamic acid.
[0083] Examples of the aliphatic acyl groups include a lower
alkanoyl group optionally having one or more suitable substituents
such as carboxy, e.g., formyl, acetyl, propionyl, butyryl,
isobutyryl, valeryl, isovaleryl, pivaloyl, hexanoyl, carboxyacetyl,
carboxypropionyl, carboxybutyryl, carboxyhexanoyl, etc.; a
cyclo(lower)alkoxy(lower)alkanoy- l group optionally having one or
more suitable substituents such as lower alkyl, e.g.,
cyclopropyloxyacetyl, cyclobutyloxypropionyl,
cycloheptyloxybutyryl, menthyloxyacetyl, menthyloxypropionyl,
menthyloxybutyryl, menthyloxypentanoyl, menthyloxyhexanoyl, etc.; a
camphorsulfonyl group; or a lower alkylcarbamoyl group having one
or more suitable substituents such as carboxy or protected carboxy,
for example, carboxy(lower)alkylcarbamoyl group (e.g.,
carboxymethylcarbamoyl, carboxyethylcarbamoyl,
carboxypropylcarbamoyl, carboxybutylcarbamoyl,
carboxypentylcarbamoyl, carboxyhexylcarbamoyl, etc.),
tri-(lower)alkylsilyl(lower)alkoxycarbonyl(lower)alkylcarbamoyl
group (e.g., trimethylsilylmethoxycarbonylethylcarbamoyl,
trimethylsilylethoxycarbonylpropylcarbamoyl,
triethylsilylethoxycarbonylp- ropylcarbamoyl,
tert-butyldimethylsilylethoxycarbonylpropylcarbamoyl,
trimethylsilylpropoxycarbonylbutylcarbamoyl, etc.) and so on.
[0084] Examples of the aromatic acyl groups include an aroyl group
optionally having one or more suitable substituents such as nitro,
e.g., benzoyl, toluoyl, xyloyl, naphthoyl, nitrobenzoyl,
dinitrobenzoyl, nitronaphthoyl, etc.; and an arenesulfonyl group
optionally having one or more suitable substituents such as
halogen, e.g., benzenesulfonyl, toluenesulfonyl, xylenesulfonyl,
naphthalenesulfonyl, fluorobenzenesulfonyl, chlorobenzenesulfonyl,
bromobenzenesulfonyl, iodobenzenesulfonyl, etc.
[0085] Examples of the aliphatic acyl groups substituted by an
aromatic group include ar(lower)alkanoyl group optionally having
one or more suitable substituents such as lower alkoxy or
trihalo(lower)alkyl, e.g., phenylacetyl, phenylpropionyl,
phenylbutyryl, 2-trifluoromethyl-2-methoxy- -2-phenylacetyl,
2-ethyl-2-trifluoromethyl-2-phenylacetyl,
2-trifluoromethyl-2-propoxy-2-phenylacetyl, etc.
[0086] More preferable acyl groups among the aforesaid acyl groups
are C.sub.1-C.sub.4 alkanoyl group optionally having carboxy,
cyclo(C.sub.5-C.sub.6)alkoxy(C.sub.1-C.sub.4)alkanoyl group having
two (C.sub.1-C.sub.4) alkyls at the cycloalkyl moiety,
camphorsulfonyl group, carboxy-(C.sub.1-C.sub.4)alkylcarbamoyl
group, tri(C.sub.1-C.sub.4)alkyls-
ilyl(C.sub.1-C.sub.4)-alkoxycarbonyl(C.sub.1-C.sub.4)alkylcarbamoyl
group, benzoyl group optionally having one or two nitro groups,
benzenesulfonyl group having halogen, or
phenyl(C.sub.1-C.sub.4)alkanoyl group having C.sub.1-C.sub.4 alkoxy
and trihalo(C.sub.1-C.sub.4)alkyl group. Among these, the most
preferable ones are acetyl, carboxypropionyl, menthyloxyacetyl,
camphorsulfonyl, benzoyl, nitrobenzoyl, dinitrobenzoyl,
iodobenzenesulfonyl and
2-trifluoromethyl-2-methoxy-2-phenylacetyl.
[0087] Preferable examples of the "5- or 6-membered nitrogen,
sulfur and/or oxygen containing heterocyclic ring" include a
pyrrolyl group and a tetrahydrofuryl group.
[0088] The ticyclic compounds (III) and its pharmaceutically
acceptable salt for use in accordance with this invention are well
known to have excellent immunosuppressive activity, antimicrobial
activity and other pharmacological activities and, as such, be of
value for the treatment or prevention of rejection reactions by
transplantation of organs or tissues, graft-vs-host diseases,
autoimmune diseases, and infectious diseases [EP-A-0184162,
EP-A-0323042, EP-A-423714, EP-A-427680, EP-A-465426, EP-A-480623,
EP-A-532088, EP-A-532089, EP-A-569337, EP-A-626385, WO89/05303,
WO93/05058, WO96/31514, WO91/13889, WO91/19495, WO93/5059,
etc.].
[0089] Particularly, the compounds which are designated as FR900506
(.dbd.FK506), FR900520 (ascomycin), FR900523, and FR900525 are
products produced by microorganisms of the genus Streptomyces, such
as Streptomyces tsukubaensis No. 9993 [deposited with National
Institute of Bioscience and Human Technology Agency of Industrial
Science and Technology (formerly Fermentation Research Institute
Agency of Industrial Science and Technology), at 1-3, Higashi
1-chome, Tsukuba-shi, Ibaraki, Japan, date of deposit Oct. 5, 1984,
accession number FERM BP-927] or Streptomyces hygroscopicus subsp.
yakushimaensis No. 7238 [deposited with National Institute of
Bioscience and Human Technology Agency of Industrial Science and
Technology (formerly Fermentation Research Institute Agency of
Industrial Science and Technology), at 1-3, Higashi 1-chome,
Tsukuba-shi, Ibaraki, Japan, date of deposit Jan. 12, 1985,
accession number FERM BP-928][EP-A-0184162]. The FK506 (general
name: tacrolimus) of the following chemical formula, in particular,
is a representative compound. 10
[0090] Chemical name:
[0091]
17-allyl-1,14-dihydroxy-12-[2-(4-hydroxy-3-methoxycyclohexyl)-1-met-
hylvinyl]-23,25-dimethoxy-13,19,21,27-tetramethyl-11,28-dioxa-4-azatricycl-
o[22.3.1.0.sup.4,9]octacos-18-ene-2,3,10,16-tetraone
[0092] The preferred examples of the tricyclic compounds (III) are
the ones, wherein each of adjacent pairs of R.sup.3 and R.sup.4 or
R.sup.5 and R.sup.6 independently form another bond formed between
the carbon atoms to which they are attached; each of R.sup.8 and
R.sup.23 is independently
[0093] a hydrogen atom;
[0094] R.sup.9 is a hydroxy group;
[0095] R.sup.10 is a methyl group, an ethyl group, a propyl group
or an allyl group;
[0096] X is (a hydrogen atom and a hydrogen atom) or an oxo
group;
[0097] Y is an oxo group;
[0098] each of R.sup.14, R.sup.15, R.sup.16, R.sup.17, R.sup.18,
R.sup.19, and R.sup.22 is a methyl group;
[0099] R.sup.24 is a 3-R.sup.20-4-R.sup.21-cyclohexyl group, in
which R.sup.20 is hydroxy, an alkoxy group, an oxo group, or a
[0100] --OCH.sub.2OCH.sub.2CH.sub.2OCH.sub.3 group, and
[0101] R.sup.21 is hydroxy, --OCN, an alkoxy group, a heteroaryloxy
which may be substituted by suitable substituents, a
--OCH.sub.2OCH.sub.2CH.sub- .2OCH.sub.3 group, a protected hydroxy
group, chloro, bromo, iodo, aminooxalyloxy, an azido group,
p-tolyloxythiocarbonyloxy, or R.sup.25R.sup.26CHCOO--,
[0102] in which R.sup.25 is optionally protected hydroxy or
protected amino, and
[0103] R.sup.26 is hydrogen or methyl, or
[0104] R.sup.20 and R.sup.21 together form an oxygen atom in an
epoxide ring; and
[0105] n is an integer of 1 or 2.
[0106] The most preferable tricyclic compounds (III) is, in
addition to FK506, ascomycin derivatives such as
halogenated-ascomycin (e.g., 33-epi-chloro-33-desoxyascomycin),
which is disclosed in EP 427,680, example 66a.
[0107] As the other preferable example of the IL-2 inhibitor,
rapamycin [THE MERCK INDEX (12th edition), No. 8288] and its
derivatives can be exemplified. Preferred example of the
derivatives is an O-substituted derivative in which the hydroxy in
position 40 of formula A illustrated at page 1 of WO 95/16691,
incorporated herein by reference, is replaced by --OR.sub.1 in
which R.sub.1 is hydroxyalkyl, hydroalkoxyalkyl, acylaminoalkyl and
aminoalkyl; for example 40-O-(2-hydroxy)ethyl-rapamyci- n,
40-O-(3-hydroxy)propyl-rapaamycin,
40-O-[2-(2-hydroxy)ethoxy]ethyl-rapa- mycin and
40-O-(2-acetaminoethyl)-rapamycin. These O-substituted derivatives
may be produced by reacting rapamycin (or dihydro or
deoxo-rapamycin) with an organic radical attached to a leaving
group (for example RX where R is the organic radical which is
desired as the O-substituent, such as an alkyl, allyl, or benzyl
moiety, and X is a leaving group such as CCl.sub.3C(NH)O or
CF.sub.3SO.sub.3) under suitable reaction conditions. The
conditions may be acidic or neutral conditions, for example in the
presence of an acid like trifluoromethanesulfonic acid,
camphorsulfonic acid, p-toluenesulfonic acid or their respective
pyridinium or substituted pyridinium salts when X is
CCl.sub.3C(NH)O or in the presence of a base like pyridine, a
substituted pyridine, diisopropylethylamine or
pentamethylpiperidine when X is CF.sub.3SO.sub.3. The most
preferable one is 40-O-(2-hydroxy)ethyl rapamycin, which is
disclosed in WO94/09010, the disclosure of which is incorporated
herein by reference.
[0108] The tricyclic compounds (III), and rapamycin and its
derivatives, may be in a form of its salt, which includes
conventional non-toxic and pharmaceutically acceptable salt such as
the salt with inorganic or organic bases, specifically, an alkali
metal salt such as sodium salt and potassium salt, an alkali earth
metal salt such as calcium salt and magnesium salt, an ammonium
salt and an amine salt such as triethylamine salt and
N-benzyl-N-methylamine salt.
[0109] With respect to the IL-2 inhibitor of the present invention,
particularly the tricyclic macrolide compounds, it is to be
understood that there may be conformers and one or more
stereoisomers such as optical and geometrical isomers due to
asymmetric carbon atom(s) or double bond(s), and such conformers
and isomers are also included within the scope of the present
invention. And further, the tricyclic macrolide compounds can be in
the form of a solvate, which is included within the scope of the
present invention. The solvate preferably include a hydrate and an
ethanolate.
[0110] Further example of the IL-2 inhibitor is cyclosporin and its
derivatives such as cyclosporin A, B, C, D, E, F, G, etc, which are
shown in THE MERCK INDEX (12th edition), No. 2821, U.S. Pat. Nos.
4,117,118, 4,215,199, 4,288,431, 4,388,307, Helv. Chim. Acta. 60,
1568(1977) and 65, 1655(1982), Transplant. Proc. 17, 1362(1985),
and so on. Among which, the most preferable one is cyclosporin A.
The disclosures of the above references are incorporated
herein.
[0111] The tricyclic compounds (III) and its pharmaceutically
acceptable salts, and cyclosporin or its derivatives may be
classified as "IL-2 production inhibitor", which show
immunosuppressive activity by inhibiting the production of IL-2.
And rapamycin or its derivatives may be classified as "IL-2 signal
transmission inhibitor", which show immunosuppressive activity by
inhibiting the transmission of IL-2 signal.
[0112] For therapeutic administration, the compound possessing the
inhibitory activity on the production of nitric oxide in the
present invention is used in the form of a conventional
pharmaceutical preparation in admixture with a conventional
pharmaceutically acceptable carrier such as an organic or inorganic
solid or liquid excipient which is suitable for oral, parenteral or
external administration. The pharmaceutical preparation may be
compounded in a solid form such as granule, capsule, tablet,
dragee, suppository or ointment, or in a liquid form such as
solution, suspension or emulsion for injection, intravenous drip,
ingestion, eye drop, etc. If needed, there may be included in the
above preparation auxiliary substance such as stabilizing agent,
wetting or emulsifying agent, buffer or any other commonly used
additives.
[0113] The "compound possessing the inhibitory activity on the
production of nitric oxide" as the effective ingredient may usually
be administered in a amount which can inhibit the production of
nitric oxide. In particular, it may be a unit dose of 0.001 mg/kg
to 500 mg/kg, preferably 0.01 mg/kg to 10 mg/kg, 1 to 4 times a
day. However, the above dosage may be increased or decreased
according to age, body weight and conditions of the patient or
administering method.
[0114] If advisable, the compound possessing an inhibitory activity
on the production of nitric oxide can be mixed with the IL-2
inhibitor prior to its use. So, the composition comprising the said
compound possessing the inhibitory activity on the production of
nitric oxide of the present invention may further comprise the IL-2
inhibitor. And optionally, it comprises further additional
ingredients, such as, mycophenolate mofetil (CellCept), steroids,
Azathiopurine, and so on.
[0115] The following Preparations and Examples are given for the
purpose of illustrating the present invention in detail.
[0116] In the following Examples and Preparations, there are
employed the other abbreviations in addition to the abbreviations
adopted by the IUPAC-IUB (Commission on Biological
Nomenclature).
[0117] The abbreviations used are as follows.
[0118] Boc: tert-butoxycarbonyl
[0119] Me: methyl
[0120] Et: ethyl
[0121] Pr: propyl
[0122] i-Pr: isopropyl
[0123] Bu: butyl
[0124] Ph: phenyl
[0125] Ts: p-toluenesulfonyl
[0126] Ac: acetyl
[0127] Bn: benzyl
[0128] Cbz: benzlyoxycarbonyl
[0129] Tf: trifluoromethanesulfonyl
[0130] The starting compounds used and the compounds obtained in
the following Preparations are given in the Tables as below, in
which the formula of the starting compounds are in the left and the
formula of the object compounds are in the right, respectively.
1 Preparation Formula No. starting compound object compound 1 11 12
2 13 14 3 15 16 4 17 18 5 19 20 6 21 22 7 23 24 8 25 26
[0131] Preparation 1
[0132] A solution of the starting compound (669 mg) and 40%
methylamine (0.7 ml) in a mixture of acetic acid (0.7 ml) and
xylene (7 ml) was refluxed for 4 hours in a flask equipped with a
Dean-Stark trap. The mixture was concentrated, neutralized with 1N
sodium hydroxide solution, and extracted three times with
chloroform. The organic layer was washed with brine, dried over
magnesium sulfate, filtered and concentrated. The residue was
purified by column chromatography (silica gel,
chloroform/methanol=50/1) to give the object compound as an oil
(445 mg).
[0133] MASS (ESI) (m/z): 288 (M+H).sup.+
[0134] .sup.1H-NMR (CDCl3,300 MHz).delta.: 1.46(9H,s), 3.60(3H,s),
4.48(2H,d,J=5 Hz), 5.33(1H,br s), 6.99(1H,s), 7.30-7.52(5H,m)
[0135] Preparation 2
[0136] To a solution of the starting compound (3.10 g) in methanol
(15 ml) was added concentrated hydrochloric acid (3 ml), and the
mixture was heated to 50.degree. C. for 2 hours. The mixture was
concentrated, made basic with a 1N sodium hydroxide solution, and
extracted three times with chloroform. The organic layer was dried
over magnesium sulfate, and filtered. Evaporation of the solvent
gave the object compound (2.35 g).
[0137] MASS (ESI) (m/z): 308 (M+H).sup.+
[0138] .sup.1H-NMR (CDCl3,300 MHz).delta.: 3.02-3.22(2H,m),
3.21(3H,s), 3.78(3H,s), 4.11 (1H,t,J=7 Hz), 6.81(2H,d,J=8 Hz),
6.99(2H,d,J=8 Hz), 7.04(1H,s), 7.21-7.48(5H,m)
[0139] Preparation 3
[0140] To an ice-cooled mixture of the starting compound (599 mg),
2-aminoacetophenone hydrochloride (362 mg) and
1-hydroxy-benzotriazole (270 mg) in dichloromethane (6 ml) was
added 1-(3-dimethylamino-propyl)-3- -ethylcarbodiimide (349 mg).
The mixture was stirred at room temperature for 12 hours. A
saturated aqueous sodium hydrogencarbonate solution was added to
the mixture, and then the mixture was extracted three times with
chloroform. The organic layer was washed with brine, dried over
magnesium sulfate, filtered, and concentrated. The residue was
purified by column chromatography (silica gel,
chloroform/methanol=70/1) to give the object compound (823 mg).
[0141] MASS (ESI) (m/z): 417 (M+H).sup.+
[0142] .sup.1H-NMR (CDCl3,300 MHz).delta.: 1.41(9H,s),
2.96-3.20(2H,m), 4.47(1H,m), 4.70(2H,AB of ABX,JAB=15 Hz),
5.01(1H,br s), 6.92(1H,br s), 7.13(2H,d,J=8 Hz), 7.24(2H,d,J=8 Hz),
7.41-7.68(3H,m), 7.88-8.00(2H,m)
[0143] Preparation 4
[0144] The object compound was obtained according to a similar
manner to that of Preparation 3.
[0145] oil
[0146] MASS: 450 (M+1)
[0147] .sup.1H-NMR (CDCl3) .delta.: 1.42(9H,s), 3.20-3.30(1H,m),
3.31-3.42 (1H,m), 4.62-4.73(1H,m), 4.70(2H,d,J=6 Hz), 6.42(1H,br
s),7.15(1H,t,J=6 Hz), 7.21(1H,d,J=6 Hz), 7.23(1H,s), 7.33(1H,s),
7.50(2H,d,J=8 Hz), 7.60(1H,t,J=8 Hz), 7.97(1H,s), 8.00(1H,br s),
8.08(2H,d,J=8 Hz), 8.57(1H,d,J=8 Hz)
[0148] Preparation 5
[0149] The object compound was obtained according to a similar
manner to that of Preparation 1.
[0150] MASS (ESI) (m/z): 473 (M+H).sup.+
[0151] .sup.1H-NMR (CDCl3,300 MHz).delta.: 0.76(3H,t,J=7 Hz),
1.38(9H,s), 1.40-1.60(2H,m), 3.48-3.80(2H,m), 3.88-4.08(2H,m),
5.40-5.60(2H,m), 7.02-7.65(10H,m), 7.92(1H,s), 8.52(1H,d,J=5
Hz)
[0152] Preparation 6
[0153] The object compound was obtained according to a similar
manner to that of Preparation 2.
[0154] MASS (ESI) (m/z): 373 (M+H).sup.+
[0155] .sup.1H-NMR (CDCl3,300 MHz).delta.: 0.78(3H,t,J=7 Hz),
1.36-1.72 (2H,m),3.42-3.74(2H,m), 3.85-4.24(2H,m), 4.81-5.02(1H,m),
7.08(1H,s), 7.15-7.72(9H,m), 7.93(1H,s), 8.55(1H,d,J=5 Hz)
[0156] Preparation 7
[0157] To an ice-cooled solution of the starting compound (100 mg),
indole-2-carboxylic acid (50 mg) and 1-hydroxybenzotriazole (41.9
mg) in dichloromethane (10 ml) was added
1-(3-dimethylaminopropyl)-3-ethylcarbod- iimide hydrochloride (71.4
mg). The mixture was stirred at room temperature for 12 hours. A
saturated aqueous sodium hydrogencarbonate solution was added to
the mixture, and then the mixture was extracted three times with
chloroform. The organic layer was washed with brine, dried over
magnesium sulfate, filtered and concentrated. The residue was
purified by column chromatography (silica gel,
chloroform/methanol=70/1) to give the object compound as white
powder (50 mg).
[0158] MASS(m/z): 466 (M+1)
[0159] .sup.1H-NMR (CDCl3) .delta.: 1.43(3H,t,J=7 Hz), 3.48(3H,s),
3.60(2H,m), 4.03(2H,q,J=7 Hz), 5.97(1H,m), 6.91(2H,d,J=8 Hz),
6.94(1H,s), 6.99(1H,s), 7.10-7.12(3H,m), 7.17(2H,d,J=8 Hz),
7.37(1H,d,J=8 Hz), 7.50(1H,t,J=8 Hz), 7.63(1H,d,J=8 Hz),
9.41(1H,s)
[0160] Preparation 8
[0161] The compound was obtained according to a similar manner to
that of Preparation 7.
[0162] MASS (ESI) (m/z): 516 (M+H).sup.+
[0163] .sup.1H-NMR (DMSO-d6,300 MHz).delta.: 0.64(3H,t,J=7 Hz),
1.31-1.55(2H,m),3.41-3.67(2H,m), 3.90-4.28(2H,m),
5.86-6.00(1H,m),6.97-7.- 21(5H,m), 7.27(1H,s), 7.29-7.42(2H,m),
7.53(2H,d,J=8 Hz), 7.55-7.68(2H,m), 7.73(2H,d,J=8 Hz), 7.81(1H,s),
8.32(1H,s), 8.49(1H,d,J=5 Hz), 9.09(1H,br d,J=8 Hz), 10.50(1H,br
s)
[0164] Preparation 9
[0165] To a solution of
2-amino-1-(4-morpholin-4-ylphenyl)ethan-1-one dihydrochloride (3.71
g), (2S)-(tert-butoxycarbonylamino)-3-(2-pyridyl)pr- opanoic acid
(5.73 g) and diphenylphosphoryl azide (3.48 g) in
N,N-dimethylformamide (70 ml) was added dropwise
N,N-diisopropylethylamin- e (4.41 ml) at 0.degree. C. and the
mixture was stirred for 20 minutes. The mixture was heated to
ambient temperature and stirred for 8 hours. The resulting mixture
was diluted with ethyl acetate (200 ml) and washed successively
with water, a saturated aqueous sodium hydrogencarbonate solution
and brine. The organic layer was dried over magnesium sulfate and
concentrated in vacuo. The residual solid was triturated with ethyl
acetate-diisopropyl ether (1:2) to give
(2S)-(tert-butoxycarbonylamino)-N-
-[2-(4-morpholin-4-ylphenyl)-2-oxoethyl]-3-(2-pyridyl)-propanamide
(2.06 g) as off-white crystals.
[0166] ESI-NS; 469 (M+H)
[0167] .sup.1H-NMR (300 MHz, CDCl3) .delta. 1.46(9H,s),
3.20-3.38(6H,m), 3.82-3.88(4H,m), 4.60(2H,d,J=5 Hz), 4.64-4.74
(1H,br), 6.37-6.45(1H,br), 6.86(2H,d,J=9 Hz), 7.14(1H,dd,J=5, 8
Hz), 7.21 (1H,d,J=8 Hz), 7.59(1H,t,J=8 Hz), 7.82-7.90(3H,m), 8.56
(1H, d, J=5 Hz)
[0168] Preparation 10
[0169] To a solution of
(2S)-(tert-butoxycarbonylamino)-N-[2-(4-morpholin--
4-ylphenyl)-2-oxoethyl]-3-(2-pyridyl)-propanamide (2.0 g) in acetic
acid (4.0 ml) and xylene (60 ml) was added methylamine (40% in
water, 4.0 ml) and the mixture was refluxed for 3 hours in a
round-bottomed flask equipped with a Dean-Stark apparatus. The
mixture was cooled to ambient temperature and a mixture of acetic
acid (4.0 ml) and methylamine (40% in water, 4.0 ml) was added to
the solution. The solution was refluxed for 2 hours and cooled to
ambient temperature. The solution was extracted with 1N
hydrochloric acid (100 ml) and the aqueous layer was washed with
ethyl acetate (50 ml). The aqueous layer was made basic with a
saturated aqueous sodium hydrogencarbonate solution and extracted
with ethyl acetate (100 ml). The organic layer was washed
successively with an aqueous sodium hydrogencarbonate solution and
brine, dried over magnesium sulfate and concentrated in vacuo. The
residue was purified by silica gel column chromatography (eluent;
2% methanol in chloroform) to give
(1S)-(tert-butoxy)-N-[1-[1-methyl-5-(4-morpholin-4-ylphenyl)imidazol-2-yl-
]-2-(2-pyridyl)ethyl]formamide (1.45 g) as yellow crystals.
[0170] ESI-MS; 464 (M+H)
[0171] .sup.1H-NMR (300 MHz, CDCl3) .delta. 1.37(9H,s),
3.17-3.23(4H,m), 3.40(3H,s), 3.41-3.47(2H,m), 3.83-3.92(4H,m),
5.33-5.47(2H,m), 6.93(1H,s), 6.94(2H,d, J=9 Hz), 7.08-7.16(2H,m),
7.21 (2H,d,J=9 Hz), 7.56(1H,t,J=8 Hz), 8.55(1H,d,J=5 Hz)
[0172] Preparation 11
[0173] To a solution of
(1S)-(tert-butoxy)-N-[1-[1-methyl-5-(4-morpholin-4-
-ylphenyl)imidazol-2-yl]-2-(2-pyridyl)ethyl] formamide (1.43 g) in
dichloromethane (25 ml) was added trifluoroacetic acid (5.0 ml) at
0.degree. C. and the mixture was stirred at ambient temperature for
2.5 hours. The resulting mixture was concentrated in vacuo and the
residue was dissolved in water (20 ml). The aqueous layer was made
basic with a saturated aqueous sodium hydrogencarbonate solution
and extracted with chloroform (80 ml). The organic layer was dried
over magnesium sulfate and concentrated in vacuo. The residue was
triturated with ethyl acetate-diisopropyl ether (1:2) to give
(1S)-[1-methyl-5-(4-morpholin-4-y-
lphenyl)imidazol-2-yl]-2-(2-pyridyl)-ethylamine (1.02 g) as yellow
crystals.
[0174] ESI-MS; 364 (M+H)
[0175] .sup.1H-NNR (300 MHz, CDCl3).delta. 3.17-3.23(4H,m),
3.27-3.47(2H,m), 3.49(3H,s), 3.84-3.91 (4H,m), 4.58(1H,dd,J=5, 8
Hz), 6.95(2H,d,J=9 Hz), 6.97(1H,s), 7.11-7.18(2H,m), 7.23(2H,d, J=9
Hz), 7.59(1H,t,J=8 Hz), 8.58(1H,d,J=5 Hz)
[0176] Preparation 12
[0177] To a solution of
(1S)-[1-methyl-5-(4-morpholin-4-ylphenyl)-imidazol-
-2-yl]-2(2-pyridyl)ethylamine (120 mg),
5-chlorobenzo[d]furan-2-carboxylic acid (68.1 mg) and
1-hydroxybenzotriazole (49.1 mg) in N,N-dimethylformamide (2.0 ml)
was added 1-(3-dimethylaminopropyl)-3-ethy- lcarbodiimide
hydrochloride (69.6 mg). The mixture was stirred at ambient
temperature for 2 hours and allowed to stand overnight. The
resulting mixture was diluted with water (20 ml) and extracted with
ethyl acetate (25 ml). The organic layer was extracted with 1N
hydrochloric acid (15 ml) and the aqueous layer was made basic with
a saturated aqueous sodium hydrogencarbonate solution, then
extracted with ethyl acetate (20 ml). The organic layer was washed
with brine, dried over magnesium sulfate and concentrated in vacuo.
The residual solid was treated with hot acetonitrile (1.5 ml) and
the mixture was cooled to ambient temperature. The solid was
collected by filtration and washed with acetonitrile to give
(1S)-(5-chlorobenzo[d]furan-2-yl)-N-[1-[1-methyl-5-(4-morpholin-4-yl-
phenyl)imidazol-2-yl]-2-(2-pyridyl)ethyl]formamide (89 mg) as
off-white crystals
[0178] mp 162-164.degree. C.
[0179] ESI-MS; 542 (M+H)
[0180] .sup.1H-NMR (300 MHz, DMSO-d.sub.6).delta. 3.11-3.17(4H,m),
3.42-3.60(2H,m), 3.53(3H,s), 3.70-3.77(4H,m), 5.83(1H,q,J=8 Hz),
6.88(1H,s), 6.99(2H,d,J=9 Hz), 7.18(1H,dd,J=5, 8 Hz), 7.26(2H,d,J=9
Hz), 7.30(1H,d,J=8 Hz), 7.47(1H,d,J=8 Hz), 7.59(1H,s),
7.63(1H,d,J=8 Hz), 7.68(1H,d,J=8 Hz), 7.87(1H,s), 8.48(1H,d,J=5
Hz), 9.32(1H,d, J=8 Hz)
[0181] [.alpha.]D=171.90 (CHCl3, c=1.030%)
[0182] Preparation 13
[0183] In order to illustrate the activity of the compounds (I) and
(II) the pharmacological test result of the representative
compounds (a). (g) of the compound (I) and (II), which were
obtained in a similar manner to that of the above-mentioned
Preparations and/or the above-identified WO 98/27108
(PCT/JP97/04243), are shown in the following.
[0184] Test Compounds: 2728
[0185] Test: Assay for inhibitory activity on the production of
nitric oxide
[0186] The murine macrophage cell line RAW264.7 (American Type
Culture Collection, No. TIB71) was used in this study. RAW264.7
cells were grown on F75 plastic culture flasks at 37.degree. C., 5%
in Dulbecco's modified Eagle's medium (DMEM) supplemented with
L-glutamine, penicillin, streptomycin and 10% heat-inactivated
fetal bovine serum. They were removed from culture flasks by rubber
cell scraper and were centrifuged and resuspended in DMEM without
phenol red. They were plated in 96-well microtiter plates (10.sup.5
cells per well) and allowed to adhere over 2 hours. The test
samples were added and the cells were preincubated for 1 hour.
Thereafter the cells were activated with both of lipopolysaccharide
(LPS) (1 .mu.g/ml) and interferon .gamma. (INF .gamma.) (3 u/ml)
for 18-24 hours. An equal volume of Griess reagent (1%
sulfanilamide/0.1% N-naphthylethylenediamine dihydrochloride/2.5%
H3PO4) was added and the cells were incubated at room temperature
for 10 minutes. The absorbance was read at 570 nm using microplate
reader and NO2- was measured using NaNO2 as a standard.
[0187] Test result:
2 Test compound (10.sup.-5 M) Inhibition (%) (a) 100 (b) 100 (c)
100 (d) 100 (e) 100 (f) 100 (g) 100
EXAMPLE 1
[0188] Protective effect of the compound (f) or (g) combined with
FK506 on rat cardiac allograft:
[0189] (Method)
[0190] Experiments were performed on male Lewis and ACI rats
weighing 175-200 g. Rats were anesthetized with sodium
pentobarbital (50 mg/kg, i.p.), and performed allogeneic (Lewis
donor to ACI recipient) heterotopic intra-abdominal cardiac
transplantation. Experimental groups were divided into single-drug
group and combined-drug groups. Single-drug dose of FK506 was 0.32
mg/kg. Combined-dose groups were K506(0.32 mg/kg)+the compound
(f)(3.2 mg/kg), FK506(0.32 mg/kg)+the compound (f) (10 mg/kg), and
FK506(0.32 mg/kg)+the compound (g) (10 mg/kg). The grafted hearts
were monitored by daily palpation and complete rejection was
defined as the cessation of palpable contractile activity. Drugs
were suspended in a solution of 0.5% methylcellulose, and
administered by daily gastric intubation in a volume of 5 ml/kg of
body weight for 14 days. The blood samples were collected on 5 days
after transplantation for analysis of nitrite/nitrate (NOx). The
concentration of NOx in rat plasma was measured by using a
spectrophotometric assay based upon the Griess reaction.
[0191] (Result)
[0192] The compounds (f) or (g), which was used in the preparation
13, was examined in combination with a potent immunosuppressive
agent (FK506) to determine whether they could improve rat cardiac
allograft survival. Graft survival and the concentration of NOx are
shown in the following tables 1 and 2.
3TABLE 1 Protective effect of the compound (f) combined with FK506
on rat cardiac allograft. Test MST NOx compound(s) n (day) (.mu.M)
FK506 12 8.5 >40 (0.32 mg/kg) Compound (f) 11 12 <30 (3.2
mg/kg) + FK506 (0.32 mg/kg) Compound (f) 12 >40### <30 (10
mg/kg) + FK506 (0.32 mg/kg) Significant vs. FK506 (0.32 mg/kg) at
###: p < 0.001 by Mann-Whitney test. Significant vs. FK506 (0.32
mg/kg) at *: p < 0.05, **: p < 0.01 by Mann-Whitney test.
MST: Median Survival Time
[0193]
4TABLE 2 Protective effect of the compound (g) combined with FK506
on rat cardiac allograft. Test MST compound n (day) FK506 6 10
(0.32 mg/kg) Compound (g) (10 mg/kg) + 9 >30 FK506 (0.32
mg/kg)
[0194] The elevated NOx production during cardiac allograft
rejection was inhibited by the compound (f) or (g) in a
dose-dependent manner. The combination of the compound (f) or (g)
and FK506 dramatically prolonged the graft survival. These results
indicate that NOx plays a pivotal role in the regulation of graft
survival and the compound (f) or (g) is good candidates for
combination with FK506 on allograft model.
[0195] The major immune mechanisms of cellular allograft rejection
are T lymphocyte-mediated cytotoxicity and delayed-type
hypersensitivity. The effector cells of delayed-type
hypersensitivity are macrophages and nitric oxide generated by iNOS
in macrophages has been demonstrated to have cytotoxicity to
allograft.
[0196] It is known that FK506 and cyclosporine A inhibit immune
reaction mediated by lymphocytes at a low dose, but they do not
strongly inhibit nitric oxide production by iNOS in
macrophages.
[0197] According to the present invention, allograft rejection
could remarkably and/or synergistically be reduced by suppression
of nitric oxide production by macrophages and further by
suppression of cytotoxic T cell.
[0198] Particularly, the present invention is quite useful for
treating and/or preventing acute rejection after
transplantation.
[0199] While the effective dosage of the IL-2 inhibitor depends on
the type of the said IL-2 inhibitor, the patient's age, type of
disease, severity of illness, and other factors, a daily dose
thereof is about 0.01.1000 mg, preferably 0.05.500 mg, and more
preferably, 0.1.100 mg for therapeutic purposes. The average unit
dose may be generally about 0.1 mg, 0.5 mg, 1 mg, 5 mg, 10 mg, 50
mg, 100 mg, 250 mg, or 500 mg.
[0200] When the above-mentioned tricyclic macrolides is used as the
IL-2 inhibitor in the present invention, the pharmaceutical
composition of the present invention is useful for increasing the
effect of the treatment and/or prevention of the following diseases
and conditions because of the pharmacologic activities possessed by
the said tricyclic macrolides.
[0201] Rejection reactions by transplantation of organs or tissues
such as the heart, kidney, liver, bone marrow, skin, cornea, lung,
pancreas, small intestine, limb, muscle, nerve, intervertebral
disc, trachea, myoblast, cartilage, etc.; graft-versus-host
reactions following bone marrow transplantation; autoimmune
diseases such as rheumatoid arthritis, systemic lupus
erythematosus, Hashimoto's thyroiditis, multiple sclerosis,
myasthenia gravis, type I diabetes, etc.; and infections caused by
pathogenic microorganisms (e.g. Aspergillus fumigatus, Fusarium
oxysporum, Trichophyton asteroides, etc.). Inflammatory or
hyperproliferative skin diseases or cutaneous manifestations of
immunologically-mediated diseases (e.g. psoriasis, atopic
dermatitis, contact dermatitis, eczematoid dermatitis, seborrheic
dermatitis, lichen planus, pemphigus, bullous pemphigoid,
epidermolysis bullosa, urticaria, angioedema, vasculitides,
erythema, dermal eosinophilia, lupus erythematosus, acne, and
alopecia areata); autoimmune diseases of the eye (e.g.
keratoconjunctivitis, vernal conjunctivitis, uveitis associated
with Behcet's disease, keratitis, herpetic keratitis, conical
keratitis, corneal epithelial dystrophy, keratoleukoma, ocular
premphigus, Mooren's ulcer, scleritis, Graves' ophthalmopathy,
Vogt-Koyanagi-Harada syndrome, keratoconjunctivitis sicca (dry
eye), phlyctenule, iridocyclitis, sarcoidosis, endocrine
ophthalmopathy, etc.); reversible obstructive airways diseases
[asthma (e.g. bronchial asthma, allergic asthma, intrinsic asthma,
extrinsic asthma, and dust asthma), particularly chronic or
inveterate asthma (e.g. late asthma and airway
hyper-responsiveness) bronchitis, etc.]; mucosal or vascular
inflammations (e.g. gastric ulcer, ischemic or thrombotic vascular
injury, ischemic bowel diseases, enteritis, necrotizing
enterocolitis, intestinal damages associated with thermal burns,
leukotriene B4-mediated diseases); intestinal
inflammations/allergies (e.g. coeliac diseases, proctitis,
eosinophilic gastroenteritis, mastocytosis, Crohn's disease and
ulcerative colitis); food-related allergic diseases with
symptomatic manifestation remote from the gastrointestinal tract
(e.g. migrain, rhinitis and eczema); renal diseases (e.g.
intestitial nephritis, Goodpasture's syndrome, hemolytic uremic
syndrome, and diabetic nephropathy); nervous diseases (e.g.
multiple myositis, Guillain-Barre syndrome, Meniere's disease,
multiple neuritis, solitary neuritis, cerebral infarction,
Alzheimer's disease, Parkinson's disease, amyotrophic lateral
sclerosis, ALS, and radiculopathy); cerebral ischemic disease,
e.g., head injury, hemorrhage in brain (e.g., subarachnoid
hemorrhage, intracerebral hemorrhage), cerebral thrombosis,
cerebral embolism, cardiac arrest, stroke, transient ischemic
attack (TIA), hypertensive encephalopathy, cerebral infarction;
endocrine diseases (e.g. hyperthyroidism, and Basedow's disease);
hematic diseases (e.g. pure red cell aplasia, aplastic anemia,
hypoplastic anemia, idiopathic thrombocytopenic purpura, autoimmune
hemolytic anemia, agranulocytosis, pernicious anemia, megaloblastic
anemia, and anerythroplasia); bone diseases (e.g. osteoporosis);
respiratory diseases (e.g. sarcoidosis, pulmonary fibrosis, and
idiopathic interstitial pneumonia); skin diseases (e.g.
dermatomyositis, leukoderma vulgaris, ichthyosis vulgaris,
photosensitivity, and cutaneous T-cell lymphoma); circulatory
diseases (e.g. arteriosclerosis, atherosclerosis, aortitis
syndrome, polyarteritis nodosa, and myocardosis); collagen diseases
(e.g. scleroderma, Wegener's granuloma, and Sjogren's syndrome);
adiposis; eosinophilic fasciitis; periodontal diseases (e.g. damage
to gingiva, periodontium, alveolar bone or substantia ossea
dentis); nephrotic syndrome (e.g. glomerulonephritis); male pattern
alopecia, alopecia senile; muscular dystrophy; pyoderma and Sezary
syndrome; chromosome abnormality-associated diseases (e.g. Down's
syndrome); Addison's disease; active oxygen-mediated diseases [e.g.
organ injury (e.g. ischemic circulation disorders of organs (e.g.
heart, liver, kidney, digestive tract, etc.) associated with
preservation, transplantation, or ischemic diseases (e.g.
thrombosis, cardial infarction, etc.)); intestinal diseases (e.g.
endotoxin shock, pseudomembranous colitis, and drug- or
radiation-induced colitis); renal diseases (e.g. ischemic acute
renal insufficiency, chronic renal failure); pulmonary diseases
(e.g. toxicosis caused by pulmonary oxygen or drugs (e.g. paracort,
bleomycin, etc.), lung cancer, and pulmonary emphysema); ocular
diseases (e.g. cataracta, iron-storage disease (siderosis bulbi),
retinitis, pigmentosa, senile plaques, vitreous scarring, corneal
alkali burn); dermatitis (e.g. erythema multiforme, linear
immunoglobulin A bullous dermatitis, cement dermatitis); and other
diseases (e.g. gingivitis, periodontitis, sepsis, pancreatitis, and
diseases caused by environmental pollution (e.g. air pollution),
aging, carcinogen, metastasis of carcinoma, and hypobaropathy)];
diseases caused by histamine release or leukotriene C4 release;
restenosis of coronary artery following angioplasty and prevention
of postsurgical adhesions; autoimmune diseases and inflammatory
conditions (e.g., primary mucosal edema, autoimmune atrophic
gastritis, premature menopause, male sterility, juvenile diabetes
mellitus, pemphigus vulgaris, pemphigoid, sympathetic ophthalmitis,
lens-induced uveitis, idiopathic leukopenia, active chronic
hepatitis, idiopathic cirrhosis, discoid lupus erythematosus,
autoimmune orchitis, arthritis (e.g. arthritis deformans), or
polychondritis); Human Immunodeficiency Virus (HIV) infection,
AIDS; allergic conjunctivitis; hypertrophic cicatrix and keloid due
to trauma, burn, or surgery.
[0202] In addition, the said tricyclic macrolides have liver
regenerating activity and/or activities of stimulating hypertrophy
and hyperplasia of hepatocytes. Therefore, the pharmaceutical
composition of the present invention is useful for increasing the
effect of the therapy and/or prophylaxis of liver diseases [e.g.
immunogenic diseases (e.g. chronic autoimmune liver diseases such
as autoimmune hepatic diseases, primary biliary cirrhosis or
sclerosing cholangitis), partial liver resection, acute liver
necrosis (e.g. necrosis caused by toxins, viral hepatitis, shock,
or anoxia), hepatitis B, non-A non-B hepatitis, hepatocirrhosis,
and hepatic failure (e.g. fulminant hepatitis, late-onset hepatitis
and "acute-on-chronic" liver failure (acute liver failure on
chronic liver diseases))].
[0203] And further, the present composition is also useful for
increasing the effect of the prevention and/or treatment of various
diseases because of the useful pharmacological activity of the said
tricyclic macrolides, such as augmenting activity of
chemotherapeutic effect, activity of cytomegalovirus infection,
anti-inflammatory activity, inhibiting activity against
peptidyl-prolyl isomerase or rotamase antimalarial activity,
antitumor activity, and so on.
[0204] The patents, patent applications and publications cited
herein are incorporated by reference.
* * * * *