U.S. patent application number 14/008112 was filed with the patent office on 2014-01-16 for pharmaceutical composition for preventing or treating macular degeneration.
This patent application is currently assigned to CATHOLIC UNIVERSITY INDUSTRY ACADEMIC COOPERATION FOUNDATION. The applicant listed for this patent is Yun-Seok Cho, Jun-Sub Choi, Choun-Ki Joo, Nack-Jeong Kim, Geun-Hyeog Lee, Hye-Sung Lee, Jin-Ha Park, Jee-Hee Suh, Jae-Sik Yang, Kyu-Yang Yi, Sung-Eun Yoo. Invention is credited to Yun-Seok Cho, Jun-Sub Choi, Choun-Ki Joo, Nack-Jeong Kim, Geun-Hyeog Lee, Hye-Sung Lee, Jin-Ha Park, Jee-Hee Suh, Jae-Sik Yang, Kyu-Yang Yi, Sung-Eun Yoo.
Application Number | 20140018402 14/008112 |
Document ID | / |
Family ID | 46932137 |
Filed Date | 2014-01-16 |
United States Patent
Application |
20140018402 |
Kind Code |
A1 |
Yi; Kyu-Yang ; et
al. |
January 16, 2014 |
PHARMACEUTICAL COMPOSITION FOR PREVENTING OR TREATING MACULAR
DEGENERATION
Abstract
The present invention provides a pharmaceutical composition for
preventing or treating macular degeneration, which comprises
benzopyran derivatives substituted with secondary amines including
imidazole or pharmaceutically acceptable salts thereof as an active
ingredient. The pharmaceutical composition of the present invention
may be used in the form of eye drops.
Inventors: |
Yi; Kyu-Yang; (Daejeon,
KR) ; Yoo; Sung-Eun; (Chungcheongnam-do, KR) ;
Kim; Nack-Jeong; (Daejeon, KR) ; Suh; Jee-Hee;
(Daejeon, KR) ; Joo; Choun-Ki; (Seoul, KR)
; Choi; Jun-Sub; (Gyeonggi-do, KR) ; Yang;
Jae-Sik; (Gyeonggi-do, KR) ; Lee; Geun-Hyeog;
(Gyeonggi-do, KR) ; Cho; Yun-Seok; (Gyeonggi-do,
KR) ; Park; Jin-Ha; (Gyeonggi-do, KR) ; Lee;
Hye-Sung; (Seoul, KR) |
|
Applicant: |
Name |
City |
State |
Country |
Type |
Yi; Kyu-Yang
Yoo; Sung-Eun
Kim; Nack-Jeong
Suh; Jee-Hee
Joo; Choun-Ki
Choi; Jun-Sub
Yang; Jae-Sik
Lee; Geun-Hyeog
Cho; Yun-Seok
Park; Jin-Ha
Lee; Hye-Sung |
Daejeon
Chungcheongnam-do
Daejeon
Daejeon
Seoul
Gyeonggi-do
Gyeonggi-do
Gyeonggi-do
Gyeonggi-do
Gyeonggi-do
Seoul |
|
KR
KR
KR
KR
KR
KR
KR
KR
KR
KR
KR |
|
|
Assignee: |
CATHOLIC UNIVERSITY INDUSTRY
ACADEMIC COOPERATION FOUNDATION
SEOUL
KR
KOREA RESEARCH INSTITUTE OF CHEMICAL TECHNOLOGY
Daejeon
KR
|
Family ID: |
46932137 |
Appl. No.: |
14/008112 |
Filed: |
March 29, 2011 |
PCT Filed: |
March 29, 2011 |
PCT NO: |
PCT/KR12/02310 |
371 Date: |
September 27, 2013 |
Current U.S.
Class: |
514/397 ;
548/311.4 |
Current CPC
Class: |
A61K 47/32 20130101;
A61K 9/08 20130101; A61P 27/02 20180101; A61K 9/10 20130101; A61K
9/0048 20130101; A61K 47/10 20130101; A61K 31/4178 20130101 |
Class at
Publication: |
514/397 ;
548/311.4 |
International
Class: |
A61K 9/00 20060101
A61K009/00; A61K 31/4178 20060101 A61K031/4178 |
Foreign Application Data
Date |
Code |
Application Number |
Mar 30, 2011 |
KR |
10-2011-002846 |
Claims
1. A method for preventing or treating macular degeneration
comprising administering an effective amount of a compound of
Formula 1 or a pharmaceutically acceptable salt thereof to a
subject in need thereof: ##STR00007## wherein, R.sup.1 represents
H, CN, NO.sub.2 or NH.sub.2, R.sup.2 represents ##STR00008##
R.sup.3 and R.sup.4 are independent each other and represent H, Cl,
Br, F, C.sub.1.about.C.sub.3 straight or branched alkyl, OR.sup.b,
CF.sub.3, OCF.sub.3, NO.sub.2 or CO.sub.2R.sup.b, R.sup.a
represents C.sub.1.about.C.sub.4 straight or branched alkyl,
R.sup.b represents H or C.sub.1.about.C.sub.3 alkyl, and *
represents a chiral center.
2. The method of claim 1, wherein the compound of Formula 1 is
selected from the group consisting of:
(2S,3S,4R)-6-nitro-4-[N-(4-chlorophenyl)-N-(1H-imidazol-2-ylmethyl)amino]-
-3-hydroxy-2-methyl-2-dimethoxymethyl-3,4-dihydro-2H-1-benzopyran;
(2S,3R,4S)-6-nitro-4-[N-(4-chlorophenyl)-N-(1H-imidazol-2-ylmethyl)amino]-
-3-hydroxy-2-methyl-2-dimethoxymethyl-3,4-dihydro-2H-1-benzopyran;
(2S,3R,4S)-6-nitro-4-[N-(4-chlorophenyl)-N-(1H-imidazol-2-ylmethyl)amino]-
-3-hydroxy-2-methyl-2-dimethoxymethyl-3,4-dihydro-2H-1-benzopyran;
(2S,3S,4R)-6-nitro-4-[N-(4-chlorophenyl)-N-(1H-imidazol-2-ylmethyl)amino]-
-3-hydroxy-2-methyl-2-dimethoxymethyl-3,4-dihydro-2H-1-benzopyran;
(2S,3S,4R)-6-nitro-4-[N-(4-trifluoromethylphenyl)-N-(1H-imidazol-2-ylmeth-
yl)amino]-3-hydroxy-2-methyl-2-dimethoxymethyl-3,4-dihydro-2H-1-benzopyran-
;
(2S,3S,4R)-6-nitro-4-[N-(4-methoxyphenyl)-N-(1H-imidazol-2-ylmethyl)amin-
o]-3-hydroxy-2-methyl-2-dimethoxymethyl-3,4-dihydro-2H-1-benzopyran;
(2S,3S,4R)-6-nitro-4-[N-(4-trifluoromethoxyphenyl)-N-(1H-imidazol-2-ylmet-
hyl)amino]-3-hydroxy-2-methyl-2-dimethoxymethyl-3,4-dihydro-2H-1-benzopyra-
n;
(2S,3S,4R)-6-nitro-4-[N-(4-bromophenyl)-N-(1H-imidazol-2-ylmethyl)amino-
]-3-hydroxy-2-methyl-2-dimethoxymethyl-3,4-dihydro-2H-1-benzopyran;
(2S,3S,4R)-6-nitro-4-[N-(2,4-dimethylphenyl)-N-(1H-imidazol-2-ylmethyl)am-
ino]-3-hydroxy-2-methyl-2-dimethoxymethyl-3,4-dihydro-2H-1-benzopyran;
(2S,3S,4R)-6-nitro-4-[N-(2-isopropylphenyl)-N-(1H-imidazol-2-ylmethyl)ami-
no]-3-hydroxy-2-methyl-2-dimethoxymethyl-3,4-dihydro-2H-1-benzopyran;
(2S,3S,4R)-6-nitro-4-[N-(2,3-dimethylphenyl)-N-(1H-imidazol-2-ylmethyl)am-
ino]-3-hydroxy-2-methyl-2-dimethoxymethyl-3,4-dihydro-2H-1-benzopyran;
(2R,3R,4S)-6-nitro-4-[N-(2,3-dimethylphenyl)-N-(1H-imidazol-2-ylmethyl)am-
ino]-3-hydroxy-2-methyl-2-dimethoxymethyl-3,4-dihydro-2H-1-benzopyran;
(2S,3R,4S)-6-nitro-4-[N-(4-bromophenyl)-N-(1H-imidazol-2-ylmethyl)amino]--
3-hydroxy-2-methyl-2-dimethoxymethyl-3,4-dihydro-2H-1-benzopyran;
(2S,3R,4S)-6-nitro-4-[N-(4-methoxyphenyl)-N-(1H-imidazol-2-ylmethyl)amino-
]-3-hydroxy-2-methyl-2-dimethoxymethyl-3,4-dihydro-2H-1-benzopyran;
(2S,3S,4R)-6-nitro-4-[N-(4-fluorophenyl)-N-(1H-imidazol-2-ylmethyl)amino]-
-3-hydroxy-2-methyl-2-dimethoxymethyl-3,4-dihydro-2H-1-benzopyran;
(2S,3S,4R)-6-nitro-4-[N-(2-methoxyphenyl)-N-(1H-imidazol-2-ylmethyl)amino-
]-3-hydroxy-2-methyl-2-dimethoxymethyl-3,4-dihydro-2H-1-benzopyran;
(2S,3R,4S)-6-nitro-4-[N-(2-isopropylphenyl)-N-(1H-imidazol-2-ylmethyl)ami-
no]-3-hydroxy-2-methyl-2-dimethoxymethyl-3,4-dihydro-2H-1-benzopyran;
(2S,3R,4S)-6-nitro-4-[N-(2-methoxyphenyl)-N-(1H-imidazol-2-ylmethyl)amino-
]-3-hydroxy-2-methyl-2-dimethoxymethyl-3,4-dihydro-2H-1-benzopyran;
(2S,3R,4S)-6-nitro-4-[N-(3-chlorophenyl)-N-(1H-imidazol-2-ylmethyl)amino]-
-3-hydroxy-2-methyl-2-dimethoxymethyl-3,4-dihydro-2H-1-benzopyran;
(2S,3S,4R)-6-nitro-4-[N-(3-chlorophenyl)-N-(1H-imidazol-2-ylmethyl)amino]-
-3-hydroxy-2-methyl-2-dimethoxymethyl-3,4-dihydro-2H-1-benzopyran;
(2S,3R,4S)-6-nitro-4-[N-(4-trifluoromethoxyphenyl)-N-(1H-imidazol-2-ylmet-
hyl)amino]-3-hydroxy-2-methyl-2-dimethoxymethyl-3,4-dihydro-2H-1-benzopyra-
n;
(2S,3S,4R)-6-cyano-4-[N-(4-chlorophenyl)-N-(1H-imidazol-2-ylmethyl)amin-
o]-3-hydroxy-2-methyl-2-dimethoxymethyl-3,4-dihydro-2H-1-benzopyran;
(2S,3R,4S)-6-amino-4-[N-(4-chlorophenyl)-N-(1H-imidazol-2-ylmethyl)amino]-
-3-hydroxy-2-methyl-2-dimethoxymethyl-3,4-dihydro-2H-1-benzopyran;
(2S,3S,4R)-6-amino-4-[N-(4-chlorophenyl)-N-(1H-imidazol-2-ylmethyl)amino]-
-3-hydroxy-2-methyl-2-dimethoxymethyl-3,4-dihydro-2H-1-benzopyran;
(2S,3S,4R)-6-amino-4-[N-(4-trifluoromethylphenyl)-N-(1H-imidazol-2-ylmeth-
yl)amino]-3-hydroxy-2-methyl-2-dimethoxymethyl-3,4-dihydro-2H-1-benzopyran-
;
(2S,3R,4S)-6-amino-4-[N-(4-trifluoromethoxyphenyl)-N-(1H-imidazol-2-ylme-
thyl)amino]-3-hydroxy-2-methyl-2-dimethoxymethyl-3,4-dihydro-2H-1-benzopyr-
an;
(2S,3R,4S)-6-amino-4-[N-(2,3-dimethylphenyl)-N-(1H-imidazol-2-ylmethyl-
)amino]-3-hydroxy-2-methyl-2-dimethoxymethyl-3,4-dihydro-2H-1-benzopyran;
(2S,3R,4S)-6-amino-4-[N-(4-methoxyphenyl)-N-(1H-imidazol-2-ylmethyl)amino-
]-3-hydroxy-2-methyl-2-dimethoxymethyl-3,4-dihydro-2H-1-benzopyran;
(2S,3R,4S)-6-amino-4-[N-(4-bromophenyl)-N-(1H-imidazol-2-ylmethyl)amino]--
3-hydroxy-2-methyl-2-dimethoxymethyl-3,4-dihydro-2H-1-benzopyran;
(2S,3S,4R)-6-amino-4-[N-(2,3-dimethylphenyl)-N-(1H-imidazol-2-ylmethyl)am-
ino]-3-hydroxy-2-methyl-2-dimethoxymethyl-3,4-dihydro-2H-1-benzopyran;
(2S,3S,4R)-6-amino-4-[N-(2-methoxyphenyl)-N-(1H-imidazol-2-ylmethyl)amino-
]-3-hydroxy-2-methyl-2-dimethoxymethyl-3,4-dihydro-2H-1-benzopyran;
(2S,3S,4R)-6-amino-4-[N-(4-methoxyphenyl)-N-(1H-imidazol-2-ylmethyl)amino-
]-3-hydroxy-2-methyl-2-dimethoxymethyl-3,4-dihydro-2H-1-benzopyran;
(2S,3S,4R)-6-amino-4-[N-(2,4-dimethylphenyl)-N-(1H-imidazol-2-ylmethyl)am-
ino]-3-hydroxy-2-methyl-2-dimethoxymethyl-3,4-dihydro-2H-1-benzopyran;
(2S,3S,4R)-6-amino-4-[N-(2-isopropylphenyl)-N-(1H-imidazol-2-ylmethyl)ami-
no]-3-hydroxy-2-methyl-2-dimethoxymethyl-3,4-dihydro-2H-1-benzopyran;
(2S,3S,4R)-6-amino-4-[N-(4-trifluoromethoxyphenyl)-N-(1H-imidazol-2-ylmet-
hyl)amino]-3-hydroxy-2-methyl-2-dimethoxymethyl-3,4-dihydro-2H-1-benzopyra-
n;
(2S,3S,4R)-6-amino-4-[N-(4-bromophenyl)-N-(1H-imidazol-2-ylmethyl)amino-
]-3-hydroxy-2-methyl-2-dimethoxymethyl-3,4-dihydro-2H-1-benzopyran;
and
(2S,3S,4R)-6-amino-4-[N-(4-fluorophenyl)-N-(1H-imidazol-2-ylmethyl)amino]-
-3-hydroxy-2-methyl-2-dimethoxymethyl-3,4-dihydro-2H-1-benzopyran.
3. The method of claim 1, wherein the compound of Formula 1 or a
pharmaceutically acceptable salt thereof is administered in the
form of eye drop formulation to be delivered to the retina.
4. The method of claim 1, wherein normal regeneration of corneal
epithelial cells is not inhibited.
5. An eye drop formulation for preventing or treating macular
degeneration comprising a compound of Formula 1 or a
pharmaceutically acceptable salt thereof as an active ingredient:
##STR00009## wherein R.sup.1, R.sup.2, R.sup.3, R.sup.4 and * are
the same as defined in claim 1.
6. The eye drop formulation for preventing or treating macular
degeneration of claim 5, in a solution form or a suspension
form.
7. The method of claim 1, comprising administering an effective
amount of
(2R,3R,4S)-6-amino-4-[N-(4-chlorophenyl)-N-(1H-imidazol-2-ylmethyl)amino]-
-3-hydroxy-2-methyl-2-dimethoxymethyl-3,4-dihydro-2H-1-benzopyran
or its pharmaceutically acceptable salt as an active
ingredient.
8. An eye drop formulation for preventing or treating macular
degeneration, comprising
(2R,3R,4S)-6-amino-4-[N-(4-chlorophenyl)-N-(1H-imidazol-2-ylmethyl)amino]-
-3-hydroxy-2-methyl-2-dimethoxymethyl-3,4-dihydro-2H-1-benzopyran
or its pharmaceutically acceptable salt as an active
ingredient.
9. The eye drop formulation for preventing or treating macular
degeneration of claim 8, having in a solution form or a suspension
form.
Description
TECHNICAL FIELD
[0001] The present invention relates to a pharmaceutical
composition for preventing or treating macular degeneration, which
comprises benzopyran derivatives substituted with secondary amines
including imidazole; or pharmaceutically acceptable salts thereof
as an active ingredient.
BACKGROUND ART
[0002] The nerve tissue located at the center of the retina of the
eye is called as a macula. The macula includes most of phtoreceptor
cells responding to light stimuli; and the appearances of objects
are focused at the center of the macula. Therefore, the macular
plays an essential role in maintaining eyesight. Age-related
macular degeneration (AMD) is a chronic disease characterized by
the degenerations in the retinal pigment epithelium and Bruch's
membrane in the macular as well as the choroidal capillary.
Anatomically, the sensory retina is located in front of the retinal
pigment epithelium. The nutrition, support, recycling, and
treatment of wastes of the sensory retina depend on the retinal
pigment epithelium. Bruch's membrane having a five-layered
structure is sandwiched between the choroid and the retinal pigment
epithelium. The innermost layer is a basal membrane of the retinal
pigment epithelium; and the outermost layer is a basal membrane of
the choroidal capillary. That is, the macular degeneration is a
degenerative disease occurred in the complex of the retinal pigment
epithelium, Bruch's membrane, and the choroidal capillary.
[0003] This disease, which occurs primarily in the ages over 50
years old, is the main cause of blindness in the population of more
than 60 years old in the Western countries; and the trend thereof
is also increasing in Korea. Although the cause of age-related
macular degeneration is not still elucidated, the risk factors
include age (especially, sharp increase is shown after the age of
75 years old), smoking (most notable environmental risk factor),
high blood pressure, obesity, genetic cause, excessive UV exposure,
low blood concentration of antioxidant, and the like.
[0004] In the macular degeneration, there are two types, i.e., dry
(non-exudative) macular degeneration and wet (exudative) macular
degeneration. The dry macular degeneration (dry AMD, non-exudative
AMD, or non-neovascular AMD) is associated with the waste formation
of yellow deposits, known as drusen, under the retina. The large
formation of drusen causes disturbing the blood flow to the retina,
especially to the macular, which leads to obscure vision, thereby
bring about visual impairment. Although the dry macular
degeneration does not cause a sudden loss of vision, it may be
developed to a wet macular degeneration. Under the retina, there
are the choroid containing a set of vessels buried within the
fibrous tissue and the pigment epithelium covering the choroid
layer. The wet macular degeneration (wet AMD, exudative AMD, or
neovascular AMD) is associated with the angiogenesis from the
choroid area under the retina. The burst of these weak neovessels
causes hemorrhage and exudation, which leads to degeneration in the
macula area of the retina, thereby bring about visual impairment.
Because the wet macular degeneration is developed very rapidly, it
is known that vision can be deteriorated in several weeks; or that
loss of vision can be caused between 2 month and 3 years.
[0005] As a therapy for macular degeneration, a photodynamic
therapy (PDT) and an injection therapy of antibody against
angiogenic growth factor are currently being used. The photodynamic
therapy is a method which comprises injecting a photosensitizer,
Visudyne, through the blood vessels, followed by irradiation of the
eye with a specific laser reactive only to the photosensitizer at
the time when the photosensitizer arrives at neovessels of the
retina, so as to selectively destroy the neovessels. However, the
photodynamic therapy causes many recurrent cases after the
treatment, which requires repetitive treatments. And also, there is
a drawback that the repetitive treatments causes damage of the
retina itself. The antibody injection therapy is a method injecting
directly into the retina an anti-VEGF antibody which inhibits the
formation and proliferation of neovessels through selectively
binding to the vascular endothelial growth factor (VEGF) critical
to the formation and progression of neovessels. As a protein drug
used in the antibody injection therapy, there are Lucentis and
Avastin. Lucentis has been approved by the FDA as a therapeutic
agent of wet macular degeneration. Although Avastin is approved to
treat cancer, it is being clinically used to treat wet AMD.
[0006] The antibody injection therapy has some drawbacks: for
example, it requires high therapeutic cost, local administration
(especially direct administration into the eye), and repeated
injections. Therefore, in terms of patients' drug compliance,
therapeutic cost, etc., there is a need for developing an
(non-injectable) eye drop formulation based on a low-molecular
weight synthetic compound.
DETAILED DESCRIPTION OF THE INVENTION
Technical Problem
[0007] The present inventors performed various researches for
developing effective compounds for preventing and treating macular
degeneration. As a result, the present inventors found that certain
benzopyran derivatives having therapeutic effects for cancer,
rheumatoid arthritis, etc. previously reported by the present
inventors can be prepared as an eye drop formulation based on a
low-molecular weight material; and usefully applied to the
prevention and treatment of macular degeneration, without injecting
directly into the affected site as in the antibody injection
therapy.
[0008] Therefore, it is an object of the present invention to
provide a pharmaceutical composition for preventing or treating
macular degeneration, which comprises a certain benzopyran
derivative as an active ingredient.
Technical Solution
[0009] According to an aspect of the present invention, there is
provided a pharmaceutical composition for preventing or treating
macular degeneration, which comprises a compound of Formula 1 or
its pharmaceutically acceptable salt as an active ingredient:
##STR00001##
[0010] wherein,
[0011] R.sup.1 represents H, CN, NO.sub.2 or NH.sub.2,
[0012] R.sup.2 represents
##STR00002##
[0013] R.sup.3 and R.sup.4 are independent each other and represent
H, Cl, Br, F, C.sub.1.about.C.sub.3 straight or branched alkyl,
OR.sup.b, CF.sub.3, OCF.sub.3, NO.sub.2 or CO.sub.2R.sup.b,
[0014] R.sup.a represents C.sub.1.about.C.sub.4 straight or
branched alkyl,
[0015] R.sup.b represents H or C.sub.1.about.C.sub.3 alkyl, and
[0016] * represents the chiral center.
[0017] In an embodiment of the present invention, there is provided
a pharmaceutical composition for preventing or treating macular
degeneration, which comprises
(2R,3R,4S)-6-amino-4-[N-(4-chlorophenyl)-N-(1H-imidazol-2-ylmethyl)amino]-
-3-hydroxy-2-methyl-2-dimethoxymethyl-3,4-dihydro-2H-1-benzopyran
or its pharmaceutically acceptable salt as an active
ingredient.
[0018] In another embodiment of the present invention, there is
provided an eye drop formulation for preventing or treating macular
degeneration, which comprises
(2R,3R,4S)-6-amino-4-[N-(4-chlorophenyl)-N-(1H-imidazol-2-ylmethyl)amino]-
-3-hydroxy-2-methyl-2-di methoxymethyl-3,4-dihydro-2H-1-benzopyran
or its pharmaceutically acceptable salt as an active ingredient.
The eye drop formulation may have a solution form or a suspension
form.
Advantageous Effects
[0019] It is newly found by the present invention that the compound
of Formula 1 or its pharmaceutically acceptable salt may be
usefully applied to the prevention and treatment of macular
degeneration, the degenerative disease of retinal choroidal
capillary. Especially, it is found by the present invention that,
when
(2R,3R,4S)-6-amino-4-[N-(4-chlorophenyl)-N-(1H-imidazol-2-ylmethyl)amino]-
-3-hydroxy-2-methyl-2-dimethoxymethyl-3,4-dihydro-2H-1-benzopyran
or its pharmaceutically acceptable salt is administered to the
cornea in the form of eye drops, it is delivered to the retina
without inhibiting normal regeneration of corneal epithelial cells.
Therefore, the compound of Formula 1, including
(2R,3R,4S)-6-amino-4-[N-(4-chlorophenyl)-N-(1H-imidazol-2-ylmethyl)amino]-
-3-hydroxy-2-methyl-2-dimethoxymethyl-3,4-dihydro-2H-1-benzopyran,
or its pharmaceutically acceptable salt can be usefully applied for
preventing or treating macular degeneration in an eye drop
formulation.
DESCRIPTION OF DRAWINGS
[0020] FIG. 1 is the results (photographs) obtained by evaluating
inhibitory effects against corneal angiogenesis in animal models
with corneal damage.
[0021] FIGS. 2 and 3 are the results (graphs) obtained by
evaluating inhibitory effects against corneal angiogenesis in
animal models with corneal damage.
[0022] FIG. 4 is the results obtained by evaluating inhibitory
effects against expression of the VEGF receptor 2 (FLK-1).
[0023] FIG. 5 is the results (photographs) obtained by evaluating
effects on normal regeneration of the corneal epithelial cells.
[0024] FIG. 6 is the results (graphs) obtained by evaluating
effects on normal regeneration of the corneal epithelial cells.
[0025] FIG. 7 is the results (photographs) obtained by evaluating
inhibitory effects against retinal angiogenesis in Brown Norway
rats.
[0026] FIG. 8 is the results (graphs) obtained by evaluating
inhibitory effects against retinal angiogenesis in Brown Norway
rats.
[0027] FIG. 9 is the results (photographs) obtained by evaluating
inhibitory effects against retinal angiogenesis in micro pigs.
[0028] FIG. 10 is the results (graphs) obtained by evaluating
inhibitory effects against retinal angiogenesis in micro pigs.
[0029] FIG. 11 is the results (photographs) obtained by evaluating
inhibitory effects of the eye drop solution and the eye drop
suspension against retinal angiogenesis.
[0030] FIG. 12 is the results (graphs) obtained by evaluating
inhibitory effects of the eye drop solution and the eye drop
suspension against retinal angiogenesis.
BEST MODE FOR CARRYING OUT THE INVENTION
[0031] There is provided a pharmaceutical composition for
preventing or treating macular degeneration, which comprises a
compound of Formula 1 or its pharmaceutically acceptable salt as an
active ingredient:
##STR00003##
[0032] wherein,
[0033] R.sup.1 represents H, CN, NO.sub.2 or NH.sub.2,
[0034] R.sup.2 represents
##STR00004##
[0035] R.sup.3 and R.sup.4 are independent each other and represent
H, Cl, Br, F, C.sub.1.about.C.sub.3 straight or branched alkyl,
OR.sup.b, CF.sub.3, OCF.sub.3, NO.sub.2 or CO.sub.2R.sup.b,
[0036] R.sup.a represents C.sub.1.about.C.sub.4 straight or
branched alkyl,
[0037] R.sup.b represents H or C.sub.1.about.C.sub.3 alkyl, and
[0038] * represents the chiral center.
[0039] In the pharmaceutical composition according to the present
invention, the compound of Formula 1 may be selected from the group
consisting of: [0040]
(2S,3S,4R)-6-nitro-4-[N-(4-chlorophenyl)-N-(1H-imidazol-2-ylmethyl)amino]-
-3-hydroxy-2-methyl-2-dimethoxymethyl-3,4-dihydro-2H-1-benzopyran;
[0041]
(2S,3R,4S)-6-nitro-4-[N-(4-chlorophenyl)-N-(1H-imidazol-2-ylmethyl)amino]-
-3-hydroxy-2-methyl-2-dimethoxymethyl-3,4-dihydro-2H-1-benzopyran;
[0042]
(2R,3R,4S)-6-nitro-4-[N-(4-chlorophenyl)-N-(1H-imidazol-2-ylmethyl)amino]-
-3-hydroxy-2-methyl-2-dimethoxymethyl-3,4-dihydro-2H-1-benzopyran;
[0043]
(2R,3S,4R)-6-nitro-4-[N-(4-chlorophenyl)-N-(1H-imidazol-2-ylmethyl)amino]-
-3-hydroxy-2-methyl-2-dimethoxymethyl-3,4-dihydro-2H-1-benzopyran;
[0044]
(2S,3S,4R)-6-nitro-4-[N-(4-trifluoromethylphenyl)-N-(1H-imidazol-2-ylmeth-
yl)amino]-3-hydroxy-2-methyl-2-dimethoxymethyl-3,4-dihydro-2H-1-benzopyran-
; [0045]
(2S,3S,4R)-6-nitro-4-[N-(4-methoxyphenyl)-N-(1H-imidazol-2-ylmeth-
yl)amino]-3-hydroxy-2-methyl-2-dimethoxymethyl-3,4-dihydro-2H-1-benzopyran-
; [0046]
(2S,3S,4R)-6-nitro-4-[N-(4-trifluoromethoxyphenyl)-N-(1H-imidazol-
-2-ylmethyl)amino]-3-hydroxy-2-methyl-2-dimethoxymethyl-3,4-dihydro-2H-1-b-
enzopyran; [0047]
(2S,3S,4R)-6-nitro-4-[N-(4-bromophenyl)-N-(1H-imidazol-2-ylmethyl)amino]--
3-hydroxy-2-methyl-2-dimethoxymethyl-3,4-dihydro-2H-1-benzopyran;
[0048]
(2S,3S,4R)-6-nitro-4-[N-(2,4-dimethylphenyl)-N-(1H-imidazol-2-ylmethyl)am-
ino]-3-hydroxy-2-methyl-2-dimethoxymethyl-3,4-dihydro-2H-1-benzopyran;
[0049]
(2S,3S,4R)-6-nitro-4-[N-(2-isopropylphenyl)-N-(1H-imidazol-2-ylmet-
hyl)amino]-3-hydroxy-2-methyl-2-dimethoxymethyl-3,4-dihydro-2H-1-benzopyra-
n; [0050]
(2S,3S,4R)-6-nitro-4-[N-(2,3-dimethylphenyl)-N-(1H-imidazol-2-yl-
methyl)amino]-3-hydroxy-2-methyl-2-dimethoxymethyl-3,4-dihydro-2H-1-benzop-
yran; [0051]
(2R,3R,4S)-6-nitro-4-[N-(2,3-dimethylphenyl)-N-(1H-imidazol-2-ylmethyl)am-
ino]-3-hydroxy-2-methyl-2-dimethoxymethyl-3,4-dihydro-2H-1-benzopyran;
[0052]
(2R,3R,4S)-6-nitro-4-[N-(4-bromophenyl)-N-(1H-imidazol-2-ylmethyl)-
amino]-3-hydroxy-2-methyl-2-dimethoxymethyl-3,4-dihydro-2H-1-benzopyran;
[0053]
(2R,3R,4S)-6-nitro-4-[N-(4-methoxyphenyl)-N-(1H-imidazol-2-ylmethy-
l)amino]-3-hydroxy-2-methyl-2-dimethoxymethyl-3,4-dihydro-2H-1-benzopyran;
[0054]
(2S,3S,4R)-6-nitro-4-[N-(4-fluorophenyl)-N-(1H-imidazol-2-ylmethyl-
)amino]-3-hydroxy-2-methyl-2-dimethoxymethyl-3,4-dihydro-2H-1-benzopyran;
[0055]
(2S,3S,4R)-6-nitro-4-[N-(2-methoxyphenyl)-N-(1H-imidazol-2-ylmethy-
l)amino]-3-hydroxy-2-methyl-2-dimethoxymethyl-3,4-dihydro-2H-1-benzopyran;
[0056]
(2R,3R,4S)-6-nitro-4-[N-(2-isopropylphenyl)-N-(1H-imidazol-2-ylmet-
hyl)amino]-3-hydroxy-2-methyl-2-dimethoxymethyl-3,4-dihydro-2H-1-benzopyra-
n; [0057]
(2R,3R,4S)-6-nitro-4-[N-(2-methoxyphenyl)-N-(1H-imidazol-2-ylmet-
hyl)amino]-3-hydroxy-2-methyl-2-dimethoxymethyl-3,4-dihydro-2H-1-benzopyra-
n; [0058]
(2R,3R,4S)-6-nitro-4-[N-(3-chlorophenyl)-N-(1H-imidazol-2-ylmeth-
yl)amino]-3-hydroxy-2-methyl-2-dimethoxymethyl-3,4-dihydro-2H-1-benzopyran-
; [0059]
(2S,3S,4R)-6-nitro-4-[N-(3-chlorophenyl)-N-(1H-imidazol-2-ylmethy-
l)amino]-3-hydroxy-2-methyl-2-dimethoxymethyl-3,4-dihydro-2H-1-benzopyran;
[0060]
(2R,3R,4S)-6-nitro-4-[N-(4-trifluoromethoxyphenyl)-N-(1H-imidazol--
2-ylmethyl)amino]-3-hydroxy-2-methyl-2-dimethoxymethyl-3,4-dihydro-2H-1-be-
nzopyran; [0061]
(2S,3S,4R)-6-cyano-4-[N-(4-chlorophenyl)-N-(1H-imidazol-2-ylmethyl)amino]-
-3-hydroxy-2-methyl-2-dimethoxymethyl-3,4-dihydro-2H-1-benzopyran;
[0062]
(2R,3R,4S)-6-amino-4-[N-(4-chlorophenyl)-N-(1H-imidazol-2-ylmethyl)amino]-
-3-hydroxy-2-methyl-2-dimethoxymethyl-3,4-dihydro-2H-1-benzopyran;
[0063]
(2S,3S,4R)-6-amino-4-[N-(4-chlorophenyl)-N-(1H-imidazol-2-ylmethyl)amino]-
-3-hydroxy-2-methyl-2-dimethoxymethyl-3,4-dihydro-2H-1-benzopyran;
[0064]
(2S,3S,4R)-6-amino-4-[N-(4-trifluoromethylphenyl)-N-(1H-imidazol-2-ylmeth-
yl)amino]-3-hydroxy-2-methyl-2-dimethoxymethyl-3,4-dihydro-2H-1-benzopyran-
; [0065]
(2R,3R,4S)-6-amino-4-[N-(4-trifluoromethoxyphenyl)-N-(1H-imidazol-
-2-ylmethyl)amino]-3-hydroxy-2-methyl-2-dimethoxymethyl-3,4-dihydro-2H-1-b-
enzopyran; [0066]
(2R,3R,4S)-6-amino-4-[N-(2,3-dimethylphenyl)-N-(1H-imidazol-2-ylmethyl)am-
ino]-3-hydroxy-2-methyl-2-dimethoxymethyl-3,4-dihydro-2H-1-benzopyran;
[0067]
(2R,3R,4S)-6-amino-4-[N-(4-methoxyphenyl)-N-(1H-imidazol-2-ylmethy-
l)amino]-3-hydroxy-2-methyl-2-dimethoxymethyl-3,4-dihydro-2H-1-benzopyran;
[0068]
(2R,3R,4S)-6-amino-4-[N-(4-bromophenyl)-N-(1H-imidazol-2-ylmethyl)-
amino]-3-hydroxy-2-methyl-2-dimethoxymethyl-3,4-dihydro-2H-1-benzopyran;
[0069]
(2S,3S,4R)-6-amino-4-[N-(2,3-dimethylphenyl)-N-(1H-imidazol-2-ylme-
thyl)amino]-3-hydroxy-2-methyl-2-dimethoxymethyl-3,4-dihydro-2H-1-benzopyr-
an; [0070]
(2S,3S,4R)-6-amino-4-[N-(2-methoxyphenyl)-N-(1H-imidazol-2-ylme-
thyl)amino]-3-hydroxy-2-methyl-2-dimethoxymethyl-3,4-dihydro-2H-1-benzopyr-
an; [0071]
(2S,3S,4R)-6-amino-4-[N-(4-methoxyphenyl)-N-(1H-imidazol-2-ylme-
thyl)amino]-3-hydroxy-2-methyl-2-dimethoxymethyl-3,4-dihydro-2H-1-benzopyr-
an; [0072]
(2S,3S,4R)-6-amino-4-[N-(2,4-dimethylphenyl)-N-(1H-imidazol-2-y-
lmethyl)amino]-3-hydroxy-2-methyl-2-dimethoxymethyl-3,4-dihydro-2H-1-benzo-
pyran; [0073]
(2S,3S,4R)-6-amino-4-[N-(2-isopropylphenyl)-N-(1H-imidazol-2-ylmethyl)ami-
no]-3-hydroxy-2-methyl-2-dimethoxymethyl-3,4-dihydro-2H-1-benzopyran;
[0074]
(2S,3S,4R)-6-amino-4-[N-(4-trifluoromethoxyphenyl)-N-(1H-imidazol--
2-ylmethyl)amino]-3-hydroxy-2-methyl-2-dimethoxymethyl-3,4-dihydro-2H-1-be-
nzopyran; [0075]
(2S,3S,4R)-6-amino-4-[N-(4-bromophenyl)-N-(1H-imidazol-2-ylmethyl)amino]--
3-hydroxy-2-methyl-2-dimethoxymethyl-3,4-dihydro-2H-1-benzopyran;
and [0076]
(2S,3S,4R)-6-amino-4-[N-(4-fluorophenyl)-N-(1H-imidazol-2-ylmethyl-
)amino]-3-hydroxy-2-methyl-2-di
methoxymethyl-3,4-dihydro-2H-1-benzopyran.
[0077] Among the compounds of Formula 1, especially preferable
compound is
(2R,3R,4S)-6-amino-4-[N-(4-chlorophenyl)-N-(1H-imidazol-2-ylmethyl)amino]-
-3-hydroxy-2-methyl-2-di methoxymethyl-3,4-dihydro-2H-1-benzopyran
or its pharmaceutically acceptable salt. Therefore, in an
embodiment of the present invention, there is provided a
pharmaceutical composition for preventing or treating macular
degeneration, which comprises
(2R,3R,4S)-6-amino-4-[N-(4-chlorophenyl)-N-(1H-imidazol-2-ylmethyl)amino]-
-3-hydroxy-2-methyl-2-dimethoxymethyl-3,4-dihydro-2H-1-benzopyran
or its pharmaceutically acceptable salt as an active
ingredient.
[0078] The compound of Formula 1 may be used as a pharmaceutically
acceptable salt, including e.g., an acid addition salt derived from
pharmaceutically acceptable free acids, an alkali metal salt
(sodium salt, potassium salt, etc.), and an alkali earth metal salt
(calcium salt, magnesium salt, etc.). The free acid includes an
inorganic acid and an organic acid. The inorganic acid includes
hydrochloric acid, hydrobromic acid, nitric acid, sulfuric acid,
perchloric acid, phosphoric acid, etc. The organic acid includes
citric acid, acetic acid, lactic acid, maleic acid, fumaric acid,
gluconic acid, methanesulfonic acid, glycolic acid, succinic acid,
tartaric acid, galacturonic acid, embonic acid, glutamic acid,
aspartic acid, oxalic acid, (D) or (L) malic acid, maleic acid,
methanesulfonic acid, ethanesulfonic acid, 4-toluenesulfonic acid,
salicylic acid, citric acid, benzoic acid, malonic acid, etc.
Examples of the pharmaceutically acceptable salt of the compound of
Formula 1 include acetate, aspartate, benzoate, besylate,
bicarbonate/carbonate, bisulfate/sulfate, borate, camsylate,
citrate, edicylate, ecylate, formate, fumarate, gluceptate,
gluconate, glucuronate, hexafluorophosphate, hibenzate,
hydrochloride/chloride, hydrobromide/bromide, hydroiodide/iodide,
isethionate, latate, malate, maleate, malonate, mesylate,
methylsulfate, naphthylate, 2-napsylate, nicotinate, nitrate,
orotate, oxalate, palmitate, famoate, phosphate/hydrogen
phosphate/dihydrogen phosphate, saccharate, stearate, succinate,
tartrate, tosylate, trifluoroacetate, aluminium salt, arginine
salt, benzathine salt, calcium salt, choline salt, diethylamine
salt, diolamine salt, glycine salt, lysine salt, magnesium salt,
meglumine salt, olamine salt, potassium salt, sodium salt,
tromethamine salt, zinc salt, etc. Among them, hydrochloride and/or
trifluoroacetate are preferable.
[0079] The pharmaceutically acceptable salt of the compound of
Formula 1 may be prepared according to conventional methods. For
example, the acid addition salt may be prepared by dissolving the
compound of Formula 1 in a water-miscible organic solvent, such as
acetone, methanol, ethanol, or acetonitrile; and then adding an
organic acid in excess or an aqueous solution of an inorganic acid,
so as to precipitate or crystallize the resulting salt.
Subsequently, the acid addition salt may be isolated by evaporating
the solvent or the excessive acid and then drying the resulting
residue; or by suction-filtering the precipitated salt.
[0080] And also, the compound of Formula 1 or its pharmaceutically
acceptable salt includes its isomers, hydrates, and solvates.
[0081] The compound of Formula 1 or its pharmaceutically acceptable
salt shows an inhibitory activity against corneal angiogenesis and
inhibitory activity against expression of corneal angiogenesis
marker, FLK-1 (see FIGS. 1 to 4). The normal healing ability
(regeneration ability) of corneal epithelial cells is not inhibited
by the pharmaceutical composition of the present invention (see
FIGS. 5 and 6). And also, the compound of Formula 1 or its
pharmaceutically acceptable salt shows excellent activity for
preventing or treating macular degeneration, a disease induced by
degeneration of the retinal choroidal capillary (see FIGS. 7 to
10). Especially, it is found by the present invention that the
compound of Formula 1 or its pharmaceutically acceptable salt may
be prepared into a topically administrable external eye drop
formulation form, which shows excellent activity for preventing or
treating macular degeneration without injecting into the affected
site (see FIGS. 11 and 12). In addition, it is found by the present
invention that, when the eye drop formulation is administered to
the cornea, the active ingredient is effectively delivered to the
retina, while only a negligible amount is absorbed into the blood
(see Table 1).
[0082] Therefore, the present invention includes, within its scope,
an eye drop formulation for preventing or treating macular
degeneration, which comprises a compound of Formula 1 or its
pharmaceutically acceptable salt as an active ingredient. In an
embodiment of the present invention, there is provided an eye drop
formulation for preventing or treating macular degeneration, which
comprises
(2R,3R,4S)-6-amino-4-[N-(4-chlorophenyl)-N-(1H-imidazol-2-ylmethyl)amino]-
-3-hydroxy-2-methyl-2-dimethoxymethyl-3,4-dihydro-2H-1-benzopyran
or its pharmaceutically acceptable salt as an active ingredient.
The eye drop formulation may have a solution form or a suspension
form.
[0083] For example, an eye drop formulation in a solution form may
comprises a solubilizer such as polyethylene glycol 400, glycerin,
etc.; a stabilizer such as EDTA etc.; a buffering agent such as
boric acid etc.; a pH controlling agent such as hydrochloric acid,
sodium hydroxide, etc., in addition to the compound of Formula 1 or
its pharmaceutically acceptable salt (for example,
(2R,3R,4S)-6-amino-4-[N-(4-chlorophenyl)-N-(1H-imidazol-2-ylmethyl)amino]-
-3-hydroxy-2-methyl-2-dimethoxymethyl-3,4-dihydro-2H-1-benzopyran
or its salt). And also, an eye drop formulation in a suspension
form may comprises a viscosity controlling agent such as
cross-linked polyvinylpyrrolidone (for example, povidone K-25)
etc.; an isotonic agent such as sodium chloride, etc.; a stabilizer
such as EDTA etc.; a buffering agent such as boric acid, Borax,
etc.; a pH controlling agent such as hydrochloric acid, sodium
hydroxide, etc., in addition to the compound of Formula 1 or its
pharmaceutically acceptable salt (for example,
(2R,3R,4S)-6-amino-4-[N-(4-chlorophenyl)-N-(1H-imidazol-2-ylmeth-
yl)amino]-3-hydroxy-2-methyl-2-dimethoxymethyl-3,4-dihydro-2H-1-benzopyran
or its salt). If necessary, the pharmaceutical composition in the
eye drop formulation form may be sterilized according to
conventional methods; or further comprise a supplement agent such
as a preservative, a hydrating agent, an emulsifier, a solubilizing
agent, a salt for controlling osmotic pressure, and/or a buffering
agent.
[0084] In the pharmaceutical composition of the present invention,
the compound of Formula 1 or its pharmaceutically acceptable salt
may be administered in an amount of typically about 0.01 to about
100 mg/day, preferably 0.03 to 80 mg/day based on adults having 70
Kg of body weight, although the amount may be changed according to
the patient's age, body weight, sex, dosage form, health condition,
severity of disease, etc. The administration may be carried out in
an appropriate interval, e.g., in a single dose or in divided doses
per day, according to the doctor's or pharmacist's instruction.
[0085] The present invention includes, within its scope, a use of a
compound of Formula 1 or its pharmaceutically acceptable salt for
the manufacture of a medicament for preventing or treating macular
degeneration:
##STR00005##
[0086] wherein, R.sup.1, R.sup.2, R.sup.3 and R.sup.4 are the same
as defined in the above; and * represents the chiral center.
[0087] In the use of the present invention, the compound of Formula
1 may be preferably
(2R,3R,4S)-6-amino-4-[N-(4-chlorophenyl)-N-(1H-imidazol-2-ylmethyl)amino]-
-3-hydroxy-2-methyl-2-dimethoxymethyl-3,4-dihydro-2H-1-benzopyran.
The medicament may have a solution form or a suspension form. In
the use of the present invention, the compound of Formula 1 or its
pharmaceutically acceptable salt may be administered in an amount
of typically about 0.01 to about 100 mg/day, preferably 0.03 to 80
mg/day based on adults having 70 Kg of body weight, although the
amount may be changed according to the patient's age, body weight,
sex, dosage form, health condition, severity of disease, etc. The
administration can be carried out in an appropriate interval, e.g.,
in a single dose or in divided doses per day, according to the
doctor's or pharmacist's instruction.
[0088] The present invention also includes, within its scope, a
method for preventing or treating macular degeneration in a
patient, which comprises administering a therapeutically effective
amount of the compound of Formula 1 or its pharmaceutically
acceptable salt to the patient in need thereof:
##STR00006##
[0089] wherein, R.sup.1, R.sup.2, R.sup.3 and R.sup.4 are the same
as defined in the above; and * represents the chiral center.
[0090] In the method for preventing or treating macular
degeneration of the present invention, the compound of Formula 1
may be preferably
(2R,3R,4S)-6-amino-4-[N-(4-chlorophenyl)-N-(1H-imidazol-2-ylmethyl)amino]-
-3-hydroxy-2-methyl-2-dimethoxymethyl-3,4-dihydro-2H-1-benzopyran.
The compound may be administered in an eye drop formulation having
a solution form or a suspension form. In the method for preventing
or treating macular degeneration of the present invention, the
compound of Formula 1 or its pharmaceutically acceptable salt may
be administered in an amount of typically about 0.01 to about 100
mg/day, preferably 0.03 to 80 mg/day based on adults with 70 Kg of
body weight, although the amount may be changed according to the
patient's age, body weight, sex, dosage form, health condition,
severity of disease, etc. The administration can be carried out in
an appropriate interval, e.g., in a single dose or in divided doses
per day, according to the doctor's or pharmacist's instruction.
[0091] The present invention will be described in further detail
with reference to the following preparation examples and examples.
These e preparation examples and examples are for illustrative
purposes only and are not intended to limit the scope of the
present invention.
Preparation Example 1
Preparation of
(2S,3S,4R)-6-nitro-4-[N-(4-chlorophenyl)-N-(1H-imidazol-2-ylmethyl)amino]-
-3-hydroxy-2-methyl-2-dimethoxymethyl-3,4-dihydro-2H-1-benzopyran
[0092] The epoxide compound,
(2S,3S,4S)-6-nitro-2-methyl-2-dimethoxymethyl-3,4-epoxy-3,4-dihydro-2H-1--
benzopyran (437 mg, 1.55 mmol) and
(4-chlorophenyl)(1H-imidazol-2-ylmethyl)amine (323 mg, 1.55 mmol)
were dissolved in acetonitrile (2 mL). To the resulting solution
was added anhydrous cobalt chloride (CoCl.sub.2) (202 mg, 1.55
mmol). The reaction mixture was stirred at 60.degree. C. for 10
hours; and then a saturated aqueous solution of NaHCO.sub.3 (5 mL)
was added to the mixture, which was extracted with ethyl acetate
(30 mL). The organic layer was washed with brine, dried over
anhydrous Na.sub.2SO.sub.4, and then concentrated under reduced
pressure. The resulting residue was purified with silica gel column
chromatography (hexane:ethyl acetate=2:1) to give the title
compound (304 mg, 40%).
[0093] .sup.1H NMR (200 MHz, CDCl.sub.3) .delta. 1.49 (s, 3H), 3.60
(s, 3H), 3.63 (s, 3H), 4.32 (m, 1H), 4.57 (s, 1H), 5.14 (br s, 1H),
6.75 (br s, 2H), 6.97 (m, 4H), 7.27 (m, 2H), 7.93 (s, 1H), 8.08 (d,
1H).
Preparation Example 2
Preparation of
(2S,3R,4S)-6-nitro-4-[N-(4-chlorophenyl)-N-(1H-imidazol-2-ylmethyl)amino]-
-3-hydroxy-2-methyl-2-dimethoxymethyl-3,4-dihydro-2H-1-benzopyran
[0094] The title compound (76 mg, 34%) was prepared in accordance
with the same procedures as in Preparation Example 1, using the
epoxide compound,
(2S,3R,4R)-6-nitro-2-methyl-2-dimethoxymethyl-3,4-epoxy-3,4-dihydro-2H-1--
benzopyran (129 mg, 0.46 mmol) and
(4-chlorophenyl)(1H-imidazol-2-ylmethyl)amine (95 mg, 0.46
mmol).
[0095] .sup.1H NMR (200 MHz, CDCl.sub.3) .delta. 1.66 (s, 3H), 3.60
(s, 3H), 3.69 (s, 3H), 3.87 (br s, 1H), 4.13 (m, 1H), 4.29 (d, 1H),
4.43 (d, 1H), 4.64 (s, 1H), 5.64 (d, 1H), 6.83 (d, 2H), 6.95 (m,
4H), 7.15 (d, 2H), 7.86 (s, 1H), 8.06 (m, 2H), 8.41 (s, 1H).
Preparation Example 3
Preparation of
(2R,3R,4S)-6-nitro-4-[N-(4-chlorophenyl)-N-(1H-imidazol-2-ylmethyl)amino]-
-3-hydroxy-2-methyl-2-dimethoxymethyl-3,4-dihydro-2H-1-benzopyran
[0096] The title compound (2.13 g, 64%) was prepared in accordance
with the same procedures as in Preparation Example 1, using the
epoxide compound,
(2R,3R,4R)-6-nitro-2-methyl-2-dimethoxymethyl-3,4-epoxy-3,4-dih-
ydro-2H-1-benzopyran (1.038 g, 3.7 mmol) and
(4-chlorophenyl)(1H-imidazol-2-ylmethyl)amine (766 mg, 3.7
mmol).
[0097] .sup.1H NMR (200 MHz, CDCl.sub.3) .delta. 1.49 (s, 3H), 3.60
(s, 3H), 4.32 (m, 1H), 4.57 (s, 1H), 5.14 (br s, 1H), 6.75 (br s,
2H), 6.97 (m, 4H), 7.27 (m, 2H), 7.93 (s, 1H), 8.08 (d, 1H).
Preparation Example 4
Preparation of
(2R,3S,4R)-6-nitro-4-[N-(4-chlorophenyl)-N-(1H-imidazol-2-ylmethyl)amino]-
-3-hydroxy-2-methyl-2-dimethoxymethyl-3,4-dihydro-2H-1-benzopyran
[0098] The title compound (269 mg, 63%) was prepared in accordance
with the same procedures as in Preparation Example 1, using the
epoxide compound,
(2R,3S,4S)-6-nitro-2-methyl-2-dimethoxymethyl-3,4-epoxy-3,4-dih-
ydro-2H-1-benzopyran (250 mg, 0.88 mmol) and
(4-chlorophenyl)(1H-imidazol-2-ylmethyl)amine (183 mg, 0.88
mmol).
[0099] .sup.1H NMR (200 MHz, CDCl.sub.3) .delta. 1.66 (s, 3H), 3.60
(s, 3H), 3.69 (s, 3H), 3.87 (br s, 1H), 4.13 (m, 1H), 4.29 (d, 1H),
4.43 (d, 1H), 4.64 (s, 1H), 5.64 (d, 1H), 6.83 (d, 2H), 6.95 (m,
4H), 7.15 (d, 2H), 7.86 (s, 1H), 8.06 (m, 2H), 8.41 (s, 1H).
Preparation Example 5
Preparation of
(2S,3S,4R)-6-nitro-4-[N-(4-trifluoromethylphenyl)-N-(1H-imidazol-2-ylmeth-
yl)amino]-3-hydroxy-2-methyl-2-dimethoxymethyl-3,4-dihydro-2H-1-benzopyran
[0100] The title compound (146 mg, 22%) was prepared in accordance
with the same procedures as in Preparation Example 1, using the
epoxide compound,
(2S,3S,4S)-6-nitro-2-methyl-2-dimethoxymethyl-3,4-epoxy-3,4-dih-
ydro-2H-1-benzopyran (356 mg, 1.26 mmol) and
(4-trifluoromethylphenyl)(1H-imidazol-2-ylmethyl)amine (305 mg,
1.26 mmol).
[0101] .sup.1H NMR (200 MHz, CDCl.sub.3) .delta. 1.51 (s, 3H), 3.60
(s, 3H), 3.61 (s, 3H), 4.32 (m, 3H), 4.57 (s, 1H), 5.14 (br s, 1H),
6.85 (m, 2H), 6.95 (m, 4H), 7.38 (d, 2H), 7.91 (s, 1H), 8.05 (dd,
2H), 8.42 (m, 1H).
Preparation Example 6
Preparation of
(2S,3S,4R)-6-nitro-4-[N-(4-methoxyphenyl)-N-(1H-imidazol-2-ylmethyl)amino-
]-3-hydroxy-2-methyl-2-dimethoxymethyl-3,4-dihydro-2H-1-benzopyran
[0102] The title compound (280 mg, 28%) was prepared in accordance
with the same procedures as in Preparation Example 1, using the
epoxide compound,
(2S,3S,4S)-6-nitro-2-methyl-2-dimethoxymethyl-3,4-epoxy-3,4-dih-
ydro-2H-1-benzopyran (591 mg, 2.10 mmol) and
(4-methoxyphenyl)(1H-imidazol-2-ylmethyl)amine (427 mg, 2.10
mmol).
[0103] .sup.1H NMR (200 MHz, CDCl.sub.3) .delta. 1.47 (s, 3H), 3.59
(d, 6H), 3.68 (s, 3H), 4.30 (m, 2H), 4.54 (m, 2H), 5.02 (d, 1H),
6.67-6.78 (m, 4H), 6.89-7.26 (m, 3H), 8.04 (m, 2H).
Preparation Example 7
Preparation of
(2S,3S,4R)-6-nitro-4-[N-(4-trifluoromethoxyphenyl)-N-(1H-imidazol-2-ylmet-
hyl)amino]-3-hydroxy-2-methyl-2-dimethoxymethyl-3,4-dihydro-2H-1-benzopyra-
n
[0104] The title compound (181 mg, 47%) was prepared in accordance
with the same procedures as in Preparation Example 1, using the
epoxide compound,
(2S,3S,4S)-6-nitro-2-methyl-2-dimethoxymethyl-3,4-epoxy-3,4-dih-
ydro-2H-1-benzopyran (200 mg, 0.71 mmol) and
(4-trifluoromethoxyphenyl)(1H-imidazol-2-ylmethyl)amine (183 mg,
0.71 mmol).
[0105] .sup.1H NMR (200 MHz, CDCl.sub.3) .delta. 1.50 (s, 3H), 3.60
(d, 6H), 4.2-4.50 (m, 2H), 4.58-5.65 (m, 2H), 5.18 (s, 1H),
6.91-6.95 (m, 7H), 8.00 (s, 1H), 8.05 (dd, 1H).
Preparation Example 8
Preparation of
(2S,3S,4R)-6-nitro-4-[N-(4-bromophenyl)-N-(1H-imidazol-2-ylmethyl)amino]--
3-hydroxy-2-methyl-2-dimethoxymethyl-3,4-dihydro-2H-1-benzopyran
[0106] The title compound (310 mg, 41%) was prepared in accordance
with the same procedures as in Preparation Example 1, using the
epoxide compound,
(2S,3S,4S)-6-nitro-2-methyl-2-dimethoxymethyl-3,4-epoxy-3,4-dih-
ydro-2H-1-benzopyran (400 mg, 1.42 mmol) and
(4-bromophenyl)(1H-imidazol-2-ylmethyl)amine (359 mg, 1.42
mmol).
[0107] .sup.1H NMR (200 MHz, CDCl.sub.3) .delta. 1.48 (s, 3H), 3.61
(d, 6H), 4.10-4.19 (m, 2H), 4.20-4.40 (m, 2H), 5.13 (s, 1H),
6.70-7.01 (m, 6H), 7.21 (s, 1H), 7.94 (s, 1H), 8.06 (dd, 1H).
Preparation Example 9
Preparation of
(2S,3S,4R)-6-nitro-4-[N-(2,4-dimethylphenyl)-N-(1H-imidazol-2-ylmethyl)am-
ino]-3-hydroxy-2-methyl-2-dimethoxymethyl-3,4-dihydro-2H-1-benzopyran
[0108] The title compound (231 mg, 33%) was prepared in accordance
with the same procedures as in Preparation Example 1, using the
epoxide compound,
(2S,3S,4S)-6-nitro-2-methyl-2-dimethoxymethyl-3,4-epoxy-3,4-dih-
ydro-2H-1-benzopyran (400 mg, 1.42 mmol) and
(2,4-dimethylphenyl)(1H-imidazol-2-ylmethyl)amine (287 mg, 1.42
mmol).
[0109] .sup.1H NMR (200 MHz, CDCl.sub.3) .delta. 1.39 (s, 3H), 2.19
(s, 3H), 2.47 (s, 3H), 3.59 (d, 6H), 4.15-4.82 (m, 5H), 6.80-6.89
(m, 5H), 7.58 (d, 1H), 7.94-7.99 (dd, 1H), 8.62 (m, 1H).
Preparation Example 10
Preparation of
(2S,3S,4R)-6-nitro-4-[N-(2-isopropylphenyl)-N-(1H-imidazol-2-ylmethyl)ami-
no]-3-hydroxy-2-methyl-2-dimethoxymethyl-3,4-dihydro-2H-1-benzopyran
[0110] The title compound (140 mg, 20%) was prepared in accordance
with the same procedures as in Preparation Example 1, using the
epoxide compound,
(2S,3S,4S)-6-nitro-2-methyl-2-dimethoxymethyl-3,4-epoxy-3,4-dih-
ydro-2H-1-benzopyran (400 mg, 1.42 mmol) and
(2-isopropylphenyl)(1H-imidazol-2-ylmethyl)amine (306 mg, 1.42
mmol).
[0111] .sup.1H NMR (200 MHz, CDCl.sub.3) .delta. 1.22-1.29 (m,
10H), 3.60 (d, 6H), 4.07-4.63 (m, 5H), 6.79-7.35 (m, 6H), 7.78 (m,
1H), 7.99 (dd, 1H), 8.61 (m, 1H)
Preparation Example 11
Preparation of
(2S,3S,4R)-6-nitro-4-[N-(2,3-dimethylphenyl)-N-(1H-imidazol-2-ylmethyl)am-
ino]-3-hydroxy-2-methyl-2-dimethoxymethyl-3,4-dihydro-2H-1-benzopyran
[0112] The title compound (253 mg, 37%) was prepared in accordance
with the same procedures as in Preparation Example 1, using the
epoxide compound,
(2S,3S,4S)-6-nitro-2-methyl-2-dimethoxymethyl-3,4-epoxy-3,4-dih-
ydro-2H-1-benzopyran (400 mg, 1.42 mmol) and
(2,3-dimethylphenyl)(1H-imidazol-2-ylmethyl)amine (287 mg, 1.42
mmol).
[0113] .sup.1H NMR (200 MHz, CDCl.sub.3) .delta. 1.39 (s, 3H), 2.17
(s, 3H), 2.41 (s, 3H), 3.61 (d, 6H), 4.26-4.74 (m, 5H), 6.76-6.95
(m, 4H), 6.98 (m, 1H), 7.58 (d, 1H), 7.95 (dd, 1H), 8.63 (d,
1H).
Preparation Example 12
Preparation of
(2R,3R,4S)-6-nitro-4-[N-(2,3-dimethylphenyl)-N-(1H-imidazol-2-ylmethyl)am-
ino]-3-hydroxy-2-methyl-2-dimethoxymethyl-3,4-dihydro-2H-1-benzopyran
[0114] The title compound (416 mg, 49%) was prepared in accordance
with the same procedures as in Preparation Example 1, using the
epoxide compound,
(2R,3R,4R)-6-nitro-2-methyl-2-dimethoxymethyl-3,4-epoxy-3,4-dih-
ydro-2H-1-benzopyran (500 mg, 1.77 mmol) and
(2,3-dimethylphenyl)(1H-imidazol-2-ylmethyl)amine (358 mg, 1.77
mmol).
[0115] .sup.1H NMR (200 MHz, CDCl.sub.3) .delta. 1.39 (s, 3H), 2.17
(s, 3H), 2.41 (s, 3H), 3.61 (d, 6H), 4.26-4.74 (m, 5H), 6.76-6.95
(m, 4H), 6.98 (m, 1H), 7.58 (d, 1H), 7.95 (dd, 1H), 8.63 (d,
1H).
Preparation Example 13
Preparation of
(2R,3R,4S)-6-nitro-4-[N-(4-bromophenyl)-N-(1H-imidazol-2-ylmethyl)amino]--
3-hydroxy-2-methyl-2-dimethoxymethyl-3,4-dihydro-2H-1-benzopyran
[0116] The title compound (570 mg, 60%) was prepared in accordance
with the same procedures as in Preparation Example 1, using the
epoxide compound,
(2R,3R,4R)-6-nitro-2-methyl-2-dimethoxymethyl-3,4-epoxy-3,4-dih-
ydro-2H-1-benzopyran (500 mg, 1.78 mmol) and
(4-bromophenyl)(1H-imidazol-2-ylmethyl)amine (450 mg, 1.78
mmol).
[0117] .sup.1H NMR (200 MHz, CDCl.sub.3) .delta. 1.48 (s, 3H), 3.61
(d, 6H), 4.10-4.19 (m, 2H), 4.20-4.40 (m, 2H), 5.13 (s, 1H),
6.70-7.01 (m, 6H), 7.21 (s, 1H), 7.94 (s, 1H), 8.06 (dd, 1H).
Preparation Example 14
Preparation of
(2R,3R,4S)-6-nitro-4-[N-(4-methoxyphenyl)-N-(1H-imidazol-2-ylmethyl)amino-
]-3-hydroxy-2-methyl-2-dimethoxymethyl-3,4-dihydro-2H-1-benzopyran
[0118] The title compound (446 mg, 86%) was prepared in accordance
with the same procedures as in Preparation Example 1, using the
epoxide compound,
(2R,3R,4R)-6-nitro-2-methyl-2-dimethoxymethyl-3,4-epoxy-3,4-dih-
ydro-2H-1-benzopyran (300 mg, 1.06 mmol) and
(4-methoxyphenyl)(1H-imidazol-2-ylmethyl)amine (216 mg, 1.06
mmol).
[0119] .sup.1H NMR (200 MHz, CDCl.sub.3) .delta. 1.47 (s, 3H), 3.59
(d, 6H), 3.68 (s, 3H), 4.30 (m, 2H), 4.54 (m, 2H), 5.02 (d, 1H),
6.67-6.78 (m, 4H), 6.89-7.26 (m, 3H), 8.04 (m, 2H).
Preparation Example 15
Preparation of
(2S,3S,4R)-6-nitro-4-[N-(4-fluorophenyl)-N-(1H-imidazol-2-ylmethyl)amino]-
-3-hydroxy-2-methyl-2-dimethoxymethyl-3,4-dihydro-2H-1-benzopyran
[0120] The title compound (650 mg, 48%) was prepared in accordance
with the same procedures as in Preparation Example 1, using the
epoxide compound,
(2S,3S,4S)-6-nitro-2-methyl-2-dimethoxymethyl-3,4-epoxy-3,4-dih-
ydro-2H-1-benzopyran (800 mg, 2.84 mmol) and
(4-fluorophenyl)(1H-imidazol-2-ylmethyl)amine (380 mg, 1.8
mmol).
[0121] .sup.1H NMR (200 MHz, CDCl.sub.3) .delta. 1.49 (s, 3H), 3.60
(d, 6H), 4.30 (m, 2H), 4.60 (m, 2H), 5.05 (m, 1H), 6.76-6.97 (m,
7H), 7.95 (s, 1H), 8.03 (dd, 1H)
Preparation Example 16
Preparation of
(2S,3S,4R)-6-nitro-4-[N-(2-methoxyphenyl)-N-(1H-imidazol-2-ylmethyl)amino-
]-3-hydroxy-2-methyl-2-dimethoxymethyl-3,4-dihydro-2H-1-benzopyran
[0122] The title compound (500 mg, 58%) was prepared in accordance
with the same procedures as in Preparation Example 1, using the
epoxide compound,
(2S,3S,4S)-6-nitro-2-methyl-2-dimethoxymethyl-3,4-epoxy-3,4-dih-
ydro-2H-1-benzopyran (500 mg, 1.78 mmol) and
(2-methoxyphenyl)(1H-imidazol-2-ylmethyl)amine (253 mg, 1.25
mmol).
[0123] .sup.1H NMR (200 MHz, CDCl.sub.3) .delta. 1.38 (s, 3H), 3.60
(d, 6H), 3.91 (s, 3H), 3.97 (m, 1H), 4.74 (d, 1H), 4.60-4.84 (m,
3H), 6.80-7.03 (m, 6H), 7.58 (m, 1H), 7.99 (dd, 1H), 8.86 (m,
1H)
Preparation Example 17
Preparation of
(2R,3R,4S)-6-nitro-4-[N-(2-isopropylphenyl)-N-(1H-imidazol-2-ylmethyl)ami-
no]-3-hydroxy-2-methyl-2-dimethoxymethyl-3,4-dihydro-2H-1-benzopyran
[0124] The title compound (72 mg, 42%) was prepared in accordance
with the same procedures as in Preparation Example 1, using the
epoxide compound,
(2R,3R,4R)-6-nitro-2-methyl-2-dimethoxymethyl-3,4-epoxy-3,4-dihydro-2H-1--
benzopyran (100 mg, 0.35 mmol) and
(2-isopropylphenyl)(1H-imidazol-2-ylmethyl)amine (75 mg, 0.35
mmol).
[0125] .sup.1H NMR (200 MHz, CDCl.sub.3) .delta. 1.22-1.29 (m,
10H), 3.60 (d, 6H), 4.07-4.63 (m, 5H), 6.79-7.35 (m, 6H), 7.78 (m,
1H), 7.99 (dd, 1H), 8.61 (m, 1H).
Preparation Example 18
Preparation of
(2R,3R,4S)-6-nitro-4-[N-(2-methoxyphenyl)-N-(1H-imidazol-2-ylmethyl)amino-
]-3-hydroxy-2-methyl-2-dimethoxymethyl-3,4-dihydro-2H-1-benzopyran
[0126] The title compound (580 mg, 67%) was prepared in accordance
with the same procedures as in Preparation Example 1, using the
epoxide compound,
(2R,3R,4R)-6-nitro-2-methyl-2-dimethoxymethyl-3,4-epoxy-3,4-dih-
ydro-2H-1-benzopyran (500 mg, 1.78 mmol) and
(2-methoxyphenyl)(1H-imidazol-2-ylmethyl)amine (231 mg, 1.78
mmol).
[0127] .sup.1H NMR (200 MHz, CDCl.sub.3) .delta. 1.38 (s, 3H), 3.60
(d, 6H), 3.91 (s, 3H), 3.97 (m, 1H), 4.74 (d, 1H), 4.60-4.84 (m,
3H), 6.80-7.03 (m, 6H), 7.58 (m, 1H), 7.99 (dd, 1H), 8.86 (m,
1H).
Preparation Example 19
Preparation of
(2R,3R,4S)-6-nitro-4-[N-(3-chlorophenyl)-N-(1H-imidazol-2-ylmethyl)amino]-
-3-hydroxy-2-methyl-2-dimethoxymethyl-3,4-dihydro-2H-1-benzopyran
[0128] The title compound (337 mg, 39%) was prepared in accordance
with the same procedures as in Preparation Example 1, using the
epoxide compound,
(2R,3R,4R)-6-nitro-2-methyl-2-dimethoxymethyl-3,4-epoxy-3,4-dih-
ydro-2H-1-benzopyran (500 mg, 1.77 mmol) and
(3-chlorophenyl)(1H-imidazol-2-ylmethyl)amine (366 mg, 1.77
mmol).
[0129] .sup.1H NMR (200 MHz, CDCl.sub.3) .delta. 1.51 (s, 3H), 3.61
(d, 6H), 4.20-4.57 (m, 2H), 4.57-4.59 (m, 2H), 5.17 (s, 1H),
6.69-6.73 (m, 3H), 6.94-7.01 (m, 4H), 7.89 (m, 1H), 8.04 (dd,
1H).
Preparation Example 20
Preparation of
(2S,3S,4R)-6-nitro-4-[N-(3-chlorophenyl)-N-(1H-imidazol-2-ylmethyl)amino]-
-3-hydroxy-2-methyl-2-dimethoxymethyl-3,4-dihydro-2H-1-benzopyran
[0130] The title compound (280 mg, 35%) was prepared in accordance
with the same procedures as in Preparation Example 1, using the
epoxide compound,
(2S,3S,4S)-6-nitro-2-methyl-2-dimethoxymethyl-3,4-epoxy-3,4-dih-
ydro-2H-1-benzopyran (450 mg, 1.6 mmol) and
(3-chlorophenyl)(1H-imidazol-2-ylmethyl)amine (232 mg, 1.1
mmol).
[0131] .sup.1H NMR (200 MHz, CDCl.sub.3) .delta. 1.51 (s, 3H), 3.61
(d, 6H), 4.20-4.57 (m, 2H), 4.57-4.59 (m, 2H), 5.17 (s, 1H),
6.69-6.73 (m, 3H), 6.94-7.01 (m, 4H), 7.89 (m, 1H), 8.04 (dd,
1H).
Preparation Example 21
Preparation of
(2R,3R,4S)-6-nitro-4-[N-(4-trifluoromethoxyphenyl)-N-(1H-imidazol-2-ylmet-
hyl)amino]-3-hydroxy-2-methyl-2-dimethoxymethyl-3,4-dihydro-2H-1-benzopyra-
n
[0132] The title compound (155 mg, 40%) was prepared in accordance
with the same procedures as in Preparation Example 1, using the
epoxide compound,
(2R,3R,4R)-6-nitro-2-methyl-2-dimethoxymethyl-3,4-epoxy-3,4-dih-
ydro-2H-1-benzopyran (200 mg, 0.71 mmol) and
(4-trifluoromethoxyphenyl)(1H-imidazol-2-ylmethyl)amine 183 mg
(0.71 mmol).
[0133] .sup.1H NMR (200 MHz, CDCl.sub.3) .delta. 1.49 (s, 3H), 3.60
(d, 6H), 4.20-4.50 (m, 2H), 4.58-5.65 (m, 2H), 5.18 (s, 1H),
6.91-6.95 (m, 7H), 7.99 (s, 1H), 8.04 (dd, 1H).
Preparation Example 22
Preparation of
(2S,3S,4R)-6-cyano-4-[N-(4-chlorophenyl)-N-(1H-imidazol-2-ylmethyl)amino]-
-3-hydroxy-2-methyl-2-dimethoxymethyl-3,4-dihydro-2H-1-benzopyran
[0134] The title compound (106 mg, 28%) was prepared in accordance
with the same procedures as in Preparation Example 1, using the
epoxide compound, (2S,3S,4S)-6-cyano-2-methyl-2-di
methoxymethyl-3,4-epoxy-3,4-dihydro-2H-1-benzopyran (210 mg, 0.8
mmol) and (4-chlorophenyl)(1H-imidazol-2-ylmethyl)amine (167 mg,
0.8 mmol).
[0135] .sup.1H NMR (200 MHz, CDCl.sub.3) .delta. 1.47 (s, 3H), 3.58
(s, 3H), 3.62 (s, 3H), 4.35 (m, 1H), 4.57 (s, 1H), 5.16 (br s, 1H),
6.81-6.93 (m, 3H), 7.17 (d, 1H), 7.38 (s, 1H), 7.51 (dd, 1H).
Preparation Example 23
Preparation of
(2R,3R,4S)-6-amino-4-[N-(4-chlorophenyl)-N-(1H-imidazol-2-ylmethyl)amino]-
-3-hydroxy-2-methyl-2-dimethoxymethyl-3,4-dihydro-2H-1-benzopyran
[0136] The nitro compound (521 mg, 1.07 mmol) prepared from
Preparation Example 3 was dissolved in methanol (3 mL); and then
10% Pd/C (50 mg) was added thereto. The mixture was hydrogenated
under 3 atmosphere pressure of H.sub.2 for 12 hours. The reaction
mixture was filtered through a Celite pad to remove a solid; and
the filtrate was concentrated. The resulting residue was purified
with silica gel column chromatography
(methanol:dichloromethane=5:95) to give the title compound (368 mg,
75%).
[0137] .sup.1H NMR (200 MHz, CDCl.sub.3) .delta. 1.42 (s, 3H), 3.61
(s, 6H), 4.27 (m, 2H), 4.42 (s, 1H), 4.52 (d, 1H), 5.24 (m, 1H),
6.29 (s, 1H), 6.58 (d, 2H), 6.70 (d, 2H), 6.98 (m, 3H), 7.41 (m,
2H).
Preparation Example 24
Preparation of
(2S,3S,4R)-6-amino-4-[N-(4-chlorophenyl)-N-(1H-imidazol-2-ylmethyl)amino]-
-3-hydroxy-2-methyl-2-dimethoxymethyl-3,4-dihydro-2H-1-benzopyran
[0138] The nitro compound (177 mg, 0.36 mmol) prepared from
Preparation Example 1 was dissolved in methanol (2 mL); and then a
0.4 M aqueous solution of Cu(OAc).sub.2 (0.38 mL, 0.15 mmol) was
added thereto. Sodium borohydride (113 mg, 3.0 mmol) was slowly
added at room temperature over 10 minutes to the reaction mixture.
The reaction mixture was stirred for an hour; and then ethyl
acetate (5 mL) was added thereto. The black precipitates were
removed by filtration; and then a saturated aqueous solution of
NaHCO.sub.3 (5 mL) was added to the filtrate. The mixture was
extracted with ethyl acetate (30 mL). The organic layer was washed
with brine, dried over anhydrous sodium sulfate, and then
concentrated in vacuo to remove the solvent. The resulting residue
was purified with silica gel column chromatography (n-hexane:ethyl
acetate=1:4) to give the title compound (58 mg, 35%).
[0139] .sup.1H NMR (200 MHz, CDCl.sub.3) .delta. 1.42 (s, 3H), 3.61
(s, 6H), 4.27 (m, 2H), 4.52 (d, 1H), 4.42 (s, 1H), 5.24 (m, 1H),
6.29 (s, 1H), 6.58 (d, 2H), 6.70 (d, 2H), 6.98 (m, 3H), 7.41 (m,
2H).
Preparation Example 25
Preparation of
(2S,3S,4R)-6-amino-4-[N-(4-trifluoromethylphenyl)-N-(1H-imidazol-2-ylmeth-
yl)amino]-3-hydroxy-2-methyl-2-dimethoxymethyl-3,4-dihydro-2H-1-benzopyran
[0140] The title compound (34 mg, 57%) was prepared in accordance
with the same procedures as in Preparation Example 24, using the
nitro compound (65 mg, 0.12 mmol) prepared from Preparation Example
5.
[0141] .sup.1H NMR (200 MHz, CDCl.sub.3) .delta. 1.38 (s, 3H), 3.60
(s, 3H), 4.06-4.85 (m, 3H), 4.41 (s, 1H), 5.06 (br s, 2H), 6.31 (s,
1H), 6.57 (d, 2H), 6.80-7.18 (m, 7H)
Preparation Example 26
Preparation of
(2R,3R,4S)-6-amino-4-[N-(4-trifluoromethoxyphenyl)-N-(1H-imidazol-2-ylmet-
hyl)amino]-3-hydroxy-2-methyl-2-dimethoxymethyl-3,4-dihydro-2H-1-benzopyra-
n
[0142] The title compound (23 mg, 24%) was prepared in accordance
with the same procedures as in Preparation Example 23, using the
nitro compound (100 mg, 0.19 mmol) prepared from Preparation
Example 21.
[0143] .sup.1H NMR (200 MHz, CDCl.sub.3) .delta. 1.50 (s, 3H), 3.60
(d, 6H), 4.20-4.50 (m, 2H), 4.59 (s, 2H), 5.18 (s, 1H), 6.30 (s,
1H), 6.60 (dd, 2H), 6.70-6.96 (m, 6H)
Preparation Example 27
Preparation of
(2R,3R,4S)-6-amino-4-[N-(2,3-dimethylphenyl)-N-(1H-imidazol-2-ylmethyl)am-
ino]-3-hydroxy-2-methyl-2-dimethoxymethyl-3,4-dihydro-2H-1-benzopyran
[0144] The title compound (19 mg, 15%) was prepared in accordance
with the same procedures as in Preparation Example 23, using the
nitro compound (135 mg, 0.28 mmol) prepared from Preparation
Example 12.
[0145] .sup.1H NMR (200 MHz, CDCl.sub.3) .delta. 1.29 (s, 3H), 2.27
(s, 3H), 2.43 (s, 3H), 3.60 (s, 6H), 4.41-4.63 (m, 5H), 6.57 (dd,
1H), 6.70-7.19 (m, 6H), 7.40 (d, 1H)
Preparation Example 28
Preparation of
(2R,3R,4S)-6-amino-4-[N-(4-methoxyphenyl)-N-(1H-imidazol-2-ylmethyl)amino-
]-3-hydroxy-2-methyl-2-dimethoxymethyl-3,4-dihydro-2H-1-benzopyran
[0146] The title compound (21 mg, 23%) was prepared in accordance
with the same procedures as in Preparation Example 23, using the
nitro compound (100 mg, 0.21 mmol) prepared from Preparation
Example 14.
[0147] .sup.1H NMR (200 MHz, CDCl.sub.3) .delta. 1.36 (s, 3H), 3.60
(d, 6H), 3.64 (s, 3H), 4.20-4.60 (m, 3H), 4.45 (s, 1H), 4.70-4.90
(m, 2H), 6.50 (m, 1H), 6.70 (dd, 1H), 6.80-7.00 (m, 6H), 7.40 (d,
1H).
Preparation Example 29
Preparation of
(2R,3R,4S)-6-amino-4-[N-(4-bromophenyl)-N-(1H-imidazol-2-ylmethyl)amino]--
3-hydroxy-2-methyl-2-dimethoxymethyl-3,4-dihydro-2H-1-benzopyran
[0148] The title compound (50 mg, 53%) was prepared in accordance
with the same procedures as in Preparation Example 23, using the
nitro compound (100 mg, 0.19 mmol) prepared from Preparation
Example 13.
[0149] .sup.1H NMR (200 MHz, CDCl.sub.3) .delta. 1.48 (s, 3H), 3.61
(d, 6H), 4.10-4.19 (m, 2H), 4.22 (s, 2H), 5.13 (s, 1H), 6.33-7.15
(m, 9H).
Preparation Example 30
Preparation of
(2S,3S,4R)-6-amino-4-[N-(2,3-dimethylphenyl)-N-(1H-imidazol-2-ylmethyl)am-
ino]-3-hydroxy-2-methyl-2-dimethoxymethyl-3,4-dihydro-2H-1-benzopyran
[0150] The title compound (35 mg, 54%) was prepared in accordance
with the same procedures as in Preparation Example 23, using the
nitro compound (70 mg, 0.14 mmol) prepared from Preparation Example
11.
[0151] .sup.1H NMR (200 MHz, CDCl.sub.3) .delta. 1.29 (s, 3H), 2.27
(s, 3H), 2.43 (s, 3H), 3.60 (s, 6H), 4.41-4.63 (m, 5H), 6.57 (dd,
1H), 6.70-7.19 (m, 6H), 7.40 (d, 1H).
Preparation Example 31
Preparation of
(2S,3S,4R)-6-amino-4-[N-(2-methoxyphenyl)-N-(1H-imidazol-2-ylmethyl)amino-
]-3-hydroxy-2-methyl-2-dimethoxymethyl-3,4-dihydro-2H-1-benzopyran
[0152] The title compound (74 mg, 66%) was prepared in accordance
with the same procedures as in Preparation Example 23, using the
nitro compound (80 mg, 0.16 mmol) prepared from Preparation Example
16.
[0153] .sup.1H NMR (200 MHz, CDCl.sub.3) .delta. 1.30 (s, 3H), 3.60
(d, 6H), 3.80 (s, 3H), 4.10-4.30 (m, 2H), 4.45 (s, 1H), 4.70-4.90
(m, 2H), 6.50 (dd, 1H), 6.70-7.00 (m, 7H), 7.40 (d, 1H).
Preparation Example 32
Preparation of
(2S,3S,4R)-6-amino-4-[N-(4-methoxyphenyl)-N-(1H-imidazol-2-ylmethyl)amino-
]-3-hydroxy-2-methyl-2-dimethoxymethyl-3,4-dihydro-2H-1-benzopyran
[0154] The title compound (74 mg, 77%) was prepared in accordance
with the same procedures as in Preparation Example 23, using the
nitro compound (103 mg, 0.21 mmol) prepared from Preparation
Example 6.
[0155] .sup.1H NMR (200 MHz, CDCl.sub.3) .delta. 1.36 (s, 3H), 3.60
(d, 6H), 3.64 (s, 3H), 4.20-4.60 (m, 3H), 4.45 (s, 1H), 4.70-4.90
(m, 2H), 6.50 (m, 1H), 6.70 (dd, 1H), 6.80-7.00 (m, 6H), 7.40 (d,
1H).
Preparation Example 33
Preparation of
(2S,3S,4R)-6-amino-4-[N-(2,4-dimethylphenyl)-N-(1H-imidazol-2-ylmethyl)am-
ino]-3-hydroxy-2-methyl-2-dimethoxymethyl-3,4-dihydro-2H-1-benzopyran
[0156] The title compound (54 mg, 67%) was prepared in accordance
with the same procedures as in Preparation Example 23, using the
nitro compound (86 mg, 0.18 mmol) prepared from Preparation Example
9.
[0157] .sup.1H NMR (200 MHz, CDCl.sub.3) .delta. 1.26 (s, 3H), 2.20
(s, 3H), 2.43 (s, 3H), 3.58 (s, 6H), 4.36-4.54 (m, 3H), 4.60 (m,
2H), 6.56 (dd, 1H), 6.70 (dd, 1H), 6.80-7.15 (m, 6H), 7.36 (d,
1H).
Preparation Example 34
Preparation of
(2S,3S,4R)-6-amino-4-[N-(2-isopropylphenyl)-N-(1H-imidazol-2-ylmethyl)ami-
no]-3-hydroxy-2-methyl-2-dimethoxymethyl-3,4-dihydro-2H-1-benzopyran
[0158] The title compound (30 mg, 73%) was prepared in accordance
with the same procedures as in Preparation Example 23, using the
nitro compound (45 mg, 0.09 mmol) prepared from Preparation Example
10.
[0159] .sup.1H NMR (200 MHz, CDCl.sub.3) .delta. 1.22-1.29 (m, 9H),
3.60 (d, 6H), 4.10-4.62 (m, 5H), 6.50-6.77 (m, 2H), 6.85-7.30 (m,
6H), 7.60 (m, 1H).
Preparation Example 35
Preparation of
(2S,3S,4R)-6-amino-4-[N-(4-trifluoromethoxyphenyl)-N-(1H-imidazol-2-ylmet-
hyl)amino]-3-hydroxy-2-methyl-2-dimethoxymethyl-3,4-dihydro-2H-1-benzopyra-
n
[0160] The title compound (34 mg, 72%) was prepared in accordance
with the same procedures as in Preparation Example 23, using the
nitro compound (50 mg, 0.10 mmol) prepared from Preparation Example
7.
[0161] .sup.1H NMR (200 MHz, CDCl.sub.3) .delta. 1.50 (s, 3H), 3.60
(d, 6H), 4.20-4.50 (m, 2H), 4.59 (s, 2H), 5.18 (s, 1H), 6.30 (s,
1H), 6.60 (dd, 2H), 6.70-6.96 (m, 6H).
Preparation Example 36
Preparation of
(2S,3S,4R)-6-amino-4-[N-(4-bromophenyl)-N-(1H-imidazol-2-ylmethyl)amino]--
3-hydroxy-2-methyl-2-dimethoxymethyl-3,4-dihydro-2H-1-benzopyran
[0162] The title compound (41 mg, 88%) was prepared in accordance
with the same procedures as in Preparation Example 23, using the
nitro compound (50 mg, 0.10 mmol) prepared from Preparation Example
8.
[0163] .sup.1H NMR (200 MHz, CDCl.sub.3) .delta. 1.48 (s, 3H), 3.61
(d, 6H), 4.10-4.19 (m, 2H), 4.22 (s, 2H), 5.13 (s, 1H), 6.33-7.15
(m, 9H).
Preparation Example 37
Preparation of
(2S,3S,4R)-6-amino-4-[N-(4-fluorophenyl)-N-(1H-imidazol-2-ylmethyl)amino]-
-3-hydroxy-2-methyl-2-dimethoxymethyl-3,4-dihydro-2H-1-benzopyran
[0164] The title compound (44 mg, 95%) was prepared in accordance
with the same procedures as in Preparation Example 23, using the
nitro compound (50 mg, 0.10 mmol) prepared from Preparation Example
15.
[0165] .sup.1H NMR (200 MHz, CDCl.sub.3) .delta. 1.49 (s, 3H), 3.60
(d, 6H), 4.30 (m, 4H), 4.98 (s, 1H), 6.33 (s, 1H), 6.55 (dd, 2H),
6.60-6.92 (m, 6H).
Example 1
Evaluation of Inhibitory Effects Against Angiogenesis in Animal
Models Having Corneal Damage
[0166] The tests were performed by using Sprague Dawley rats (8
weeks old, male) as animals having corneal damage (15 rats for the
experiment group; and 1 rat for normal group, respectively). The
corneal damage was induced by applying a cotton swab coated with
silver nitrate to the surface of the rat corneas, as as to induce a
damage having a diameter of 2 mm in the central cornea. On the
third day after inducing the corneal damages, we confirmed the
establishment of corneal angiogenesis-inducing models and then
subconjunctivally injected a solution (20 .mu.l) prepared by
dissolving the compound of Preparation Example 23, i.e.,
(2R,3R,4S)-6-amino-4-[N-(4-chlorophenyl)-N-(1H-imidazol-2-ylmethyl)amino]-
-3-hydroxy-2-methyl-2-dimethoxymethyl-3,4-dihydro-2H-1-benzopyran
(KR-31831) in phosphate buffered saline (PBS) containing 10% PEG
400, in the concentration of 0.3 mg/ml. In case of the control
group, the PBS (20 .mu.l) containing 10% PEG 400 was
subconjunctivally injected. As a positive control, a solution (20
.mu.l) prepared by dissolving Avastin (used as an angiogenesis
inhibitor) in PBS in the concentration of 10 mg/ml was
subconjunctivally injected. 1 week after the drug administrations,
the photographs of the corneal angiogenesis were shown in FIG. 1.
The evaluations quantifying the photographs of FIG. 1 were
performed by using the Image J (NIH, USA) program and the results
thereof were shown in FIG. 2.
[0167] As shown in FIG. 1, the animal models injected with the
compound of the present invention showed lower corneal angiogenesis
than the control group injected with PBS and the positive group
injected with Avastin. And also, as shown in FIG. 2, the group
administered with the compound of the present invention (KR) and
the group administered with Avastin (Ava) respectively showed
52.6.+-.10% and 66.4.+-.10.6% of corneal angiogenesis, in
comparison with the corneal angiogenesis (100%) of the control
group (Cont). Therefore, it can be seen that, even when the
compound of the present invention was administered in lower
concentration than Avastin, it has superior inhibitory effect
against corneal angiogenesis (FIG. 2).
Example 2
Evaluation of Inhibitory Effects Against Angiogenesis in Animals
Model Having Corneal Damage
[0168] Except for performing both the subconjunctival injection and
the eye drop-administration of
(2R,3R,4S)-6-amino-4-[N-(4-chlorophenyl)-N-(1H-imidazol-2-ylmethyl)amino]-
-3-hydroxy-2-methyl-2-dimethoxymethyl-3,4-dihydro-2H-1-benzopyran
and Avastin, the inhibitory effects against angiogenesis were
evaluated according to the same manners as in Example 1. For the
subconjunctival injection, we used a solution (20 .mu.l) prepared
by dissolving
(2R,3R,4S)-6-amino-4-[N-(4-chlorophenyl)-N-(1H-imidazol-2-ylmethyl)amino]-
-3-hydroxy-2-methyl-2-dimethoxymethyl-3,4-dihydro-2H-1-benzopyran
in PBS containing 10% PEG 400, in the concentration of 0.3 mg/ml;
and a solution (20 .mu.l) prepared by dissolving Avastin in PBS in
the concentration of 10 mg/ml. And also, for the eye
drop-administration, we used each solution (50 .mu.l, one drop)
prepared by dissolving each drug in PBS containing 30% by weight of
PEG (polyethylene glycol 400) and 10% by weight of DMSO (dimethyl
sulfoxide) in the same concentrations thereof as mentioned in the
above; and then adjusting the pH of the resulting solution to pH 7
with hydrochloric acid. The results thereof are shown in FIG.
3.
[0169] As shown in FIG. 3, in comparison with the angiogenesis
(100%) of the control group (Cont), 29.3.+-.8.6% of angiogenesis
was shown in case of the group administered with Avastin (Ava); and
25.1.+-.10.8% of angiogenesis was shown in case of the group
administered with the compound of the present invention (KR). It
can be seen that, even when the compound of the present invention
was used in lower concentration than Avastin, it has superior
inhibitory effect against angiogenesis (FIG. 3), which is the same
as in Example 1.
Example 3
Measurement of the Expression of a VEGF Receptor 2 (FLK-1)
[0170] The tests were performed for confirming corneal angiogenesis
as a marker. To a Trizole.TM. solution (200 .mu.l), were added each
corneal tissue obtained from the rats of Example 1. Total RNAs were
isolated and then cDNAs were prepared by using a reverse
transcriptase. Each cDNA (5 .mu.l) was mixed with the primer set of
FLK-1 and then PCR amplication was performed in a thermocycler
under the following conditions; denaturation at 94.degree. C. for 5
minutes, and then 35 cycles of denaturation at 94.degree. C. for 20
seconds, annealing at 58.degree. C. for 1 minute, and extension at
72.degree. C. for 25 seconds. Each PCR product was loaded on 1%
agarose gel; and the resulting ethidium bromide fluorescent bands
were identified. The results thereof were shown in FIG. 4.
[0171] As shown in FIG. 4, it can be seen that the group (KR)
subconjunctivally injected with a solution (20 .mu.l) of the
compound of the present invention, i.e.,
(2R,3R,4S)-6-amino-4-[N-(4-chlorophenyl)-N-(1H-imidazol-2-ylmethyl)amino]-
-3-hydroxy-2-methyl-2-dimethoxymethyl-3,4-dihydro-2H-1-benzopyran
in the concentration of 0.3 mg/ml showed remarkably low (about 30%)
FLK-1 expression in the cornea, in comparison with the FLK-1
expression in the cornea (100%) in case of the control group (Non)
injected with PBS (20 .mu.l) and the FLK-1 expression in the cornea
(about 59%) in case of the group (Avastin) injected with Avastin
(20 .mu.l) in the concentration of 10 mg/ml
Example 4
Evaluation of Effects on Normal Regeneration of Corneal Epithelial
Cells in Animal Models
[0172] Animal models having damage in the corneal epithelial cells
were established by anesthetizing rats (8 weeks old, 200 g to 250
g) with a mixed solution of rompun and zoletil (1 ml/kg) and then
scraping the corneal epithelial cells at the center of the cornea
in a circular form having 2 mm diameter with a surgical knife. The
removal of corneal epithelial cells was confirmed with a 1%
fluorescein solution. The eye drop formulation (50 .mu.l) used in
Example 2 was administered twice a day, i.e., every 12 hours. The
corneas at 0 hour, 18 hours, 36 hours after the administration were
shown in FIG. 5. The evaluations quantifying the photographs of
FIG. 5 were performed by using the Image J (NIH, USA) program and
the results thereof were shown in FIG. 6.
[0173] As shown in FIGS. 5 and 6, when each corneal epithelial
damage size was compared with the whole cornea, the control group
(Control) showed 51.+-.9% of healing rate at 0 hour after the
administration, 54.+-.10% of healing rate at 18 hours after the
administration, and 92.+-.6% of healing rate at 36 hours after the
administration, respectively; and the group (KR eyedrop)
administered with the eye drop formulation containing the compound
of the present invention, i.e.,
(2R,3R,4S)-6-amino-4-[N-(4-chlorophenyl)-N-(1H-imidazol-2-ylmethyl)amino]-
-3-hydroxy-2-methyl-2-dimethoxymethyl-3,4-dihydro-2H-1-benzopyran,
showed 52.+-.9% of healing rate at 0 hour after the administration,
55.+-.9% of healing rate at 18 hours after the administration, and
94.+-.8% of healing rate at 36 hours after the administration,
respectively. Therefore, it can be seen that the normal healing
rate of corneal epithelial cells is not inhibited by the eye drop
formulation containing the compound of the present invention.
Example 5
Observation of Therapeutic Effects of Macular Degeneration in
Animal Models Having Retinal Damage
[0174] The tests were performed by using Brown Norway rats (8 weeks
old, male) as animals having retinal damage (15 rats for the
experiment group; and 1 rat for normal group, respectively). The
retinal Bruch's membrane of each Brown Norway rat was damaged using
a Zeiss 532s laser at the intensity of 200 mV, 30 ms. On the third
day after the laser-damaging, the drug was administered into the
vitreous humor in order to observe the retinal vessels. The
compound of the present invention, i.e.,
(2R,3R,4S)-6-amino-4-[N-(4-chlorophenyl)-N-(1H-imidazol-2-ylmethyl)amino]-
-3-hydroxy-2-methyl-2-dimethoxymethyl-3,4-dihydro-2H-1-benzopyran
was administered with the solutions (10 .mu.l of each solution)
obtained by dissolving in PBS containing 30% PEG 400 and 10% DMSO,
in the concentrations of 0.1 mg/ml and 0.3 mg/ml. Avastin was
administered with the solution (10 .mu.l) obtained by dissolving in
PBS in the concentration of 2.5 mg/ml. In case of the control
group, the PBS (10 .mu.l) was administered. In addition,
(2R,3R,4S)-6-amino-4-[N-(4-chlorophenyl)-N-(1H-imidazol-2-ylmethyl)amino]-
-3-hydroxy-2-methyl-2-dimethoxymethyl-3,4-dihydro-2H-1-benzopyran
was intraperitoneally administered with the solution obtained by
dissolving in PBS containing 30% PEG 400 and 10% DMSO, in the dose
of 25 mg/kg. 2 weeks after the drug administrations, dextran-FITC
was injected into the left ventricle in the dose of 25 mg/kg,
followed by enucleating the eye after 5 minutes therefrom; and then
fixing in a 10% formalin solution. The cornea and the eye lens were
removed from the fixed eye, which was then observed under a
fluorescence microscope. Angiogenesis was identified by the
intensity and leakage of dextran-FITC in the laser-induced damage
site; and the results thereof were shown in FIG. 7. The evaluations
quantifying the photographs of FIG. 7 were performed by using the
Image J (NIH, USA) program and the results thereof were shown in
FIG. 8.
[0175] As shown in FIG. 7, a reticulate form were observed in the
normal retinal vessels (Normal) by the dextran-FITC staining, while
the leakage of dextran-FITC in the laser-induced damage site was
observed in case of the control group (Control). However, it can be
seen that, in case of the group administered with the compound of
the present invention, the leakage area of dextran-FITC in the
laser-induced damage site was reduced in comparison with the
control group and the Avastin-administered group.
[0176] FIG. 8 shows the evaluation results through quantifying the
fluorescence intensities in the photographs of FIG. 7. When the
fluorescence intensity of the surrounding vessels was 100, the
control group (Control) administered with a sterile saline showed
67.+-.3 of fluorescence intensity; and the group administered with
Avastin (Ava) showed 46.+-.3 of fluorescence intensity. In
contrast, among the groups administered with the compound of the
present invention, the 0.1 mg vitreous humor-administered group
(0.1 mg/ml, VI) showed 53.+-.2.8 of fluorescence intensity; the 0.3
mg vitreous humor-administered group (0.3 mg/ml, VI) showed
45.+-.3.1 of fluorescence intensity; and the intraperitoneally
administered group (25 mg/kg, IP) showed 51.+-.5 of fluorescence
intensity. Therefore, it can be seen that the groups administered
with the compound of the present invention showed remarkably low
fluorescence intensity; and that, even when the compound of the
present invention was injected in lower concentration than Avastin
known as an angiogenesis inhibitor, it has equivalent or more
effects. In addition, from the results that the intraperitoneal
administration also showed the therapeutic effects, it can be seen
that the efficiency in terms of administration method is higher in
comparison with the conventional therapeutic agent.
Example 6
Measurement of Therapeutic Effects of Macular Degeneration in
Animal Models Having Retinal Damage
[0177] The tests were performed by using Yutacan Micro pigs (3 kg,
male) as animal models having retinal damage (15 pigs for the
experiment group). The retinal macular degeneration models were
established according to the same procedures as in Brown Norway
rats of Example 5. The eye drop formulation (100 .mu.l, 0.9 mg/ml)
prepared according to the same procedures as in Example 2 was
administered four times per day for 2 weeks. For vitreous
injection, the solution of the same compound (20 .mu.l, 0.3 mg/ml)
was injected into the vitreous humor once. 2 weeks after the
administrations, 1% fluorescein was intravenously injected so as to
observe the laser-induced damage site; and the effects of the
compound of the present invention, i.e.,
(2R,3R,4S)-6-amino-4-[N-(4-chlorophenyl)-N-(1H-imidazol-2-ylmethyl)amino]-
-3-hydroxy-2-methyl-2-dimethoxymethyl-3,4-dihydro-2H-1-benzopyran.
The results were shown in FIG. 9. The evaluations quantifying the
photographs of FIG. 9 were performed by using the Image J (NIH,
USA) program and the results thereof were shown in FIG. 10.
[0178] As shown in FIGS. 9 and 10, when the intensity of the
non-drug administered control group was 1, the eye drop
formulation-administered group (4 times per day, 2 weeks) (Eye
drop) showed 0.46.+-.0.17 of intensity; and the
vitreously-administered group (Vitreous Inj.) showed 0.36.+-.0.14
of intensity. Therefore, the compound of the present invention is
effective for the treatment of macular degeneration. Especially, it
can be seen that, even when the compound of the present invention
is administered in the eye drop formulation form, it shows an
excellent therapeutic effect of macular degeneration.
Example 7
Measurement of Retinal Delivery of Eye Drop Formulation in Animal
Models
[0179] We evaluated whether the eyd drop formulation containing
(2R,3R,4S)-6-amino-4-[N-(4-chlorophenyl)-N-(1H-imidazol-2-ylmethyl)amino]-
-3-hydroxy-2-methyl-2-dimethoxymethyl-3,4-dihydro-2H-1-benzopyran
is penetrated through the cornea and then delivered to the retina.
Brown Norway rats (8 weeks old) were used in the test; and the
animal models were established according to the same procedures as
in Example 5. The eye drop formulations (100 .mu.l a of each
formulation) of the following Formulation Examples 1 to 3 (i.e.,
the eye drop formulations in the solution form of Formulation
Examples 1 and 2; and the eye drop formulation in the suspension
form of Formulation Example 3) were administered four times per day
for 2 weeks, respectively. 2 weeks after the administrations, 1%
fluorescein was intravenously injected so as to observe the
laser-induced damage site; and the effects of the compound of the
present invention, i.e.,
(2R,3R,4S)-6-amino-4-[N-(4-chlorophenyl)-N-(1H-imidazol-2-ylmethyl)amino]-
-3-hydroxy-2-methyl-2-dimethoxymethyl-3,4-dihydro-2H-1-benzopyran.
The results were shown in FIG. 11. The evaluations quantifying the
photographs of FIG. 11 were performed by using the Image J (NIH,
USA) program and the results thereof were shown in FIG. 12.
[0180] As shown in FIGS. 11 and 12, when the intensity of the
non-drug administered control group was 100%, the group
administered with the eye drop formulation in the solution form
having the concentration of 0.9 mg/ml (Eyedrop 0.9 mg/ml) showed
64.3.+-.9.7% of intensity; the group administered with the eye drop
formulation in the solution form having the concentration of 0.6
mg/ml (Eyedrop 0.6 mg/ml) showed 75.8.+-.11.6% of intensity; and
the group administered with the eye drop formulation in the
suspension form having the concentration of 0.6 mg/ml (Suspension
0.6 mg/ml) showed 71.57.+-.11.58% of intensity. Therefore, it can
be seen that, even when the eye drop formulations prepared
according to the present invention were externally and topically
administered, those show excellent therapeutic effects of macular
degeneration.
Example 8
Measurement of Retinal Delivery of Eye Drop Formulation in Animal
Models
[0181] We evaluated whether the eyd drop formulation containing
(2R,3R,4S)-6-amino-4-[N-(4-chlorophenyl)-N-(1H-imidazol-2-ylmethyl)amino]-
-3-hydroxy-2-methyl-2-dimethoxymethyl-3,4-dihydro-2H-1-benzopyran
is penetrated through the cornea and then delivered to the retina.
Rats (8 weeks old) were used in the test; and the eye drop
formulation (50 .mu.l) prepared in the following Formulation
Example 1 was dropped on each rat's eye. The delivery level of the
compound to the retina was measured; and the results thereof are
shown in the following table 1.
TABLE-US-00001 TABLE 1 Time Aqueous humor Vitreous humor Retina
Plasma 30 minutes 1250 ng/ml 195 ng/ml 846 ng/ml N.D. 120 minutes
827 ng/ml 194 ng/ml 225 ng/ml 0.9 ng/ml N.D.: not detected
[0182] As shown in the above table 1, the compound was detected in
the levels of 1250 ng/ml in the aqueous humor, 195 ng/ml in the
vitreous humor, and 846 ng/ml in the retina, respectively, 30
minutes after the administration. 120 minutes after the
administration, the compound was detected in the levels of 827
ng/ml in the aqueous humor, 194 ng/ml in the vitreous humor, and
225 ng/ml in the retina, respectively. In the plasma, the compound
was not detected, 30 minutes after the administration; and was
detected in the level of 0.9 ng/ml, 120 minutes after the
administration. Therefore, it can be seen that, when the compound
of the present invention was dropped on the eye, it is delivered to
the retina; and that only a negligible amount is absorbed into the
blood.
[0183] The compound of Formula 1 or its salt according to the
present invention may be formulated into various dosage forms
according to the purposes, preferably into an eye drop formulation
form. The eye drop formulation may be in a solution form or in a
suspension form. The following Formulation Examples illustrate
representative examples of the eye drop formulations in a solution
or suspension form, but the present invention is not limited
thereto.
Formulation Examples 1 and 2
Preparation of Eye Drop Formulations in the Solution Form
[0184] The eye drop formulations in the solution form were prepared
according to the components and amounts shown in Table 2. Each
amount of Table 2 represents the amount thereof per 1 ml of total
volume. The compound of Preparation Example 23, polyethylene glycol
400, glycerin, EDTA, and boric acid were dissolved in sterile
water. The pH was adjusted to 6.5.+-.0.5 with diluted hydrochloric
acid; and then sterile water was added to the solution, so as to
adjust the total volume.
TABLE-US-00002 TABLE 2 Formulation Formulation Component Example 1
Example 2 The compound of 0.9 mg 0.6 mg Formula 1 (Preparation
Example 23) Polyethylene glycol 400 150 mg 150 mg Glycerin 120 mg
120 mg EDTA 0.5 mg 0.5 mg Boric acid 10 mg 10 mg Diluted
hydrochloric acid q.s. q.s.
Formulation Example 3
Preparation of Eye Drop Formulation in the Suspension Form
[0185] The eye drop formulation in the suspension form was prepared
according to the components and amounts shown in Table 3. Each
amount of Table 3 represents the amount thereof per 1 ml of total
volume. The compound of Preparation Example 23, povidone K-25,
EDTA, boric acid, Borax, and sodium chloride were dispersed in
sterile water. The pH was adjusted to 7.0.+-.0.5 with diluted
hydrochloric acid; and then sterile water was added to the
suspension, so as to adjust the total volume.
TABLE-US-00003 TABLE 3 Formulation Component Example 3 The compound
of Formula 1 0.6 mg (Preparation Example 23) Povidone K-25 20 mg
EDTA 0.5 mg Boric acid 10 mg Borax 1 mg Sodium chloride 2 mg
Diluted hydrochloric acid q.s.
* * * * *