Title of Invention

A COMPOUND OF BENZOPYRAN DERIVATIVES SUBSTITUED WITH A BENZIMIDAZOLE DERIVATIVE AND PHARMACETICALL ACCEPTABLE SALTS THEREOF

Abstract The present invention relates to benzopyran derivatives substituted with a benzimidazole derivative, or pharmaceutically acceptable salts thereof, a preparation method of the same and phamnarceutical compositions containing them. Benzopyran derivatives substituted with a benzimidazole derivative, represented in Formula (1), have the function of protecting heart from ischemia-reperfusion without side effect like vasodilation, so that a pharmaceutical composition containing benzopyran derivatives substituted with a benzimidazole derivative or pharmaceutically acceptable salts thereof of the present invention as an effective ingredient can be effectively used for the protection of tissues influenced by ischemia-reperfusion, for example, for the protection of heart, nervous cells, brain, retinal cells, storage organs, etc. and for the treatment of diseases caused by ischemia-reperfusion.
Full Text BENZOPYRAN DERIVATIVES SUBSTITUTED WL'.
BENZIMIDAZOLE DERIVATIVE, PHABMACEOTK
ACCEPTABLE SALTS THEREOF, THEIR PREPARATIONS
AND PHARMACEUTICAL COMPOSITIONS CONTAINING THEM
FIELD OF THE INVENTION
The present invention relates to benzopyran
derivatives substituted with a benzimidazole derivative,
represented in , pharmaceutically acceptable
salts thereof, processes for preparing the same and a
pharmaceutical compositions containing them as an
effective ingredient having the function of protecting
heart against damage caused by ischemia-reperfu£ ion.
(Wherein, R1, R2, R3, R4, R5, R
6
defined in the description.)
are as
BACKGROUND
ischemia heart disease results from myocardial
ischemia developed by a serious deficiency of oxygen
supply caused by interruption of blood flow to heart by
a reason like arteriosclerosis (G. J. Grover, Can. J.
Physiol. 75, 309, 1997; G. D. Lopaschuk fit al.
& Medicine 42, 1997) . Myocardial ischemia induces
pathological changes in cells progressively, leading to
irreversible myocardial damage and even necrosis of
cells and tissues, at last. In early stage when damage
is reversible, irreversible damage might be prevented
by reperfusion through surgical operations such c.s PTCA
(percutaneous transluminal coronary angioplast;
cience
CABG (coronary artery bypass graft) or
thrombolytics, but the restoration of f 1 DW by
reperfusion therapy is accompanied by a ::urther
injurious phenomenon called reperfusion injury (D. J.
Hearse, Medicographia 18, 22, 1996) . It is difficult
to clearly separate ischemic injury from that mediated
by reperfusion. Reperfusion injury is caused by sudden
restoration of blood flow by reperfusion t'lerapy,
mainly due to reactive oxygen free radicals and -alcium
overload. Reperfusion injury includes a range of
) and
using
events, such as arrhythmia, vascular damage, myocarcial
dysfunction and serious neurocognitive dysfunction.
In order to delay damage by ischemia and minimize
reperfusion injury, studies have actively been
undergoing on pharmacotherapy using immune modulators,
agents to suppress apoptosis, ion channel modulators,
etc, artificial blood products to enhance the oxygen
carrying potential of blood, and development of devices
and operation procedures, but neither of them has oeen
in commercial use, so far. As an ion cha
modulators, an inhibitor of Na-H exchanger (NHE)
adenosine Ai/A2 antagonist and a KATP opener (ATPsensitive
potassium channel opener) draw our attention.
According to earlier reports, diazoxide, a
mel
an
KATp
opener, can reduce damage due to oxidative stress by
suppressing the generation of oxygen free radicals in
mitochondria by inducing oxidation of flavoprotein (A.
A. Starkov, Biosci, Rep. 17, 273, 1997; V. P. Skulachev,
Q. Rev. Biophus. 29, 169, 1996), and the opening of
relates to the generation of antioxidant enzymes
Okubo et al., Mol. and cell Biochem, 196, 3, 1999)
the decrease of release of excitatory amino acids
Moreau, G. Huber, Brain Res., 31, 65, 1999).
general KATP openers have not only cardioprotective
activity but also vasorelaxant activity, meaning that
^•ATP
(S.
and
(J-L
The
the relaxation of coronary and peripheral blood vessels
drops blood pressure, so that blood flow to damaged
tissues decreases, which is negative factor for
cardioprotection. That is, vasorelaxation is a kind of
side effect of those openers for heart protection.
KATP, which was first found in myocardium, is
distributed in variety of organs and tissues such as
(3 -cells of pancreas, smooth muscles, kidney and
central nervous system, etc., so that it has been a
major target for the development of a novel d::ug but,
at the same time, it is hard to develop
medicine working selectively toward a specific
tissue. According to Atwal et al, the cardiopr
activity and vasorelaxant activity of K.•ATP
related each other and benzopyranyl cyanoguanidines
(BMS-180448) having a structure of
responses specifically to KATP in heart,
conventional potassium channel openers,
compounds have been confirmed to have compe
weak vasorelaxant activity, so that they can
heart without a significant hypotensive action, which
provides a new chance for the development of
therapeutic agent for ischemic heart diseases.
a novel
organ or
otective
are not
unlike
Those
ratively
protect
a novel

(Figure Removed)
Thus, the inventors of the present invention
synthesized benzopyran derivatives substituted with
benzimidazole derivatives, in which the guanidinyl
group substituted in the 4-position of benzopyran was
cyclized to a benzene ring to form a benzimidazole ring.
And the present inventors completed this invention by
confirming that the compound of the invention had an
excellent cardioprotective effect against the damage
I
caused by ischemia-reperfusion, so that it can be
effectively used as a protective agent or therc.peutic
agent for ischemia-reperfusion related diseases.
Precisely, the compound can be used for the treatment
of ischemic heart diseases such as myocardial
infarction, unstable angina pectoris, etc. and for the
protection of heart upon thrombolytic therapy or
reperfusion therapy such as PTCA (percutaneous
transluminal coronary angioplasty) and CABG (coronary
artery bypass graft), and for the protection of
ischemia-reperfusion related tissues such as nerve
cells, brain, retinal cells, storage organs, etc.
SUMMARY OF THE INVENTION
It is an object of this invention to provide
benzopyran derivatives substituted with benzimidazole
derivatives, represented in , or
pharmaceutically acceptable salts thereof.
It is also an object of this invention to provide
processes for preparing benzopyran derivatives
substituted with benzimidazole derivatives, represented
in , or pharmaceutically acceptable
thereof.
It is a further object of this invention to
provide pharmaceutical composition con :aining
benzopyran derivatives substituted with benzimjidazole
derivatives, represented in pharmaceutically acceptable salts of the same
effective ingredient.
DETAILED DESCRIPTION OF PREFERRED EMBODIMEN
In order to achieve the above object, the present
invention provides benzopyran derivatives substituted
salts
or
as an
rs
with a benzimidazole derivative, pharmaceut
acceptable salts thereof, processes for preparir
same and a pharmaceutical composition containing
as an effective ingredient.
Hereinafter, the present invention is described
in detail.
The present invention provides benz|opyran
derivatives substituted with benzimidazole derive]tives,
represented in , or pharmaceut
acceptable salts thereof.
.cally
g the
them
ically
(Wherein,
X is O, S or NCN;
\l is N02, NH2, H, CN, NHCOCH3, NHCQCF3 or
NHS02CH3;
OR' C H C H^ Z
R2 is XOR , CH2ORa or CH3
Wherein,
Ra is Ci~ C4 straight or branched alkyl;
Z is C2~ C6 straight or branched alkyl;
R3 is OH or OCOCH3;
R4 is Ci ~ C4 straight or branched alkyl;
R5 and R6 are independently H, Ci~ C4 straight or
branched alkyl, alkoxy or halogen;
* represents a chiral carbon.)
The present invention also provides, in addition
to benzopyran derivatives represented in
and pharmaceutically acceptable salts, solvat^s and
hydrates thereof.
Benzopyran derivatives of the present in1/ention
represented in include not only a ::acemic
mixture but also any diastereoisomer in which at. least
one carbon in the 2, 3, or 4-position is chiral. In
, if all the carbons in the 2, 3 and 4-
position are chiral, 3,4-dihydro benzopyran compounds
of the present invention are in the form of
diastereoisomers as seen in di), (I2) , (Is), ajid (I4)
in the below .

(Wherein, X, R1, R2, R3, R4, R5 and R6 (are as
defined in .)
Preferable compounds of include:
1) (2S, 3R, 4S)-6-nitro-3,4-dihydro-3-hydrdxy-2-
dimethoxymethyl-2-methyl-4-(2,3-dihydro-2-cyanoiminolH-
benzimidazol-l-yl)-2H-l-benzopyran;
2) (2R, 3R, 4S)-6-nitro-3,4-dihydro-3-hydrd>xy-2-
dimethoxymethyl-2-methyl-4-(2,3-dihydro-2-cyanoimi)nolH-
benzimidazol-l-yl)-2H-l-benzopyran;
3) (25, 35, 4R)-6-nitro-3,4-dihydro-3-hydrcfcxy-2-
dimethoxymethyl-2-methyl-4-(2,3-dihydro-2-cyanoim:.nolH-
benzimidazol-l-yl)-2H-l-benzopyran;
4) (2R, 3S, 4S)-6-nitro-3,4-dihydro-3-hydrbxy-2-
dimethoxymethyl-2-methyl-4-(2,3-dihydro-2-cyanoim
IH-benzimidazol-l-yl)-2H-l-benzopyran;
5) (2S, 3R, 4S)-6-amino-3,4-dihydro-3-hydrbxy-2-
dimethoxymethyl-2-methyl-4- (2, 3-dihydro-2-cyanoimi.no-
IH-benziraidazol-l-yl)-2H-l-benzopyran;
6) (25, 3R, 4S)-6-nitro-3,4-dihydro-3-hydroxy-2-
dimethoxymethyl-2-methyi-4-(2,3-dihydro-3-methyl-2-
cyanoimino-lH-benzimidazol-1-yl)-2H-l-benzopyran;
no-
7) (2S, 3R, 4S)-6-nitro-3,4-dihydro-3-hydr
dimethoxymethyl-2-methyl-4-(2,3-dihydro-5,6-dimet
cyanoimino-lH-benzimidazol-1-yl)-2H-l-benzopyran;
8) (2S, 3R, 4S)-6-amino-3,4-dihydro-3-hydroxy-2-
dimethoxymethyl-2-methyl-4-(2,3-dihydro-5,6-dimet
cyanoimino-lH-benzimidazol-l-yl)-2H-l-benzopyran;
:xy-2-
iyl-2-
hyl-2-
9) (2S, 3R, 4S)-6-nitro-3,4-dihydro-3-hydr dimethoxymethyl-2-methyl-4-(2,3-dihydro-3,5, 6-
trimethyl-2-cyanoimino-lH-benzimidazol-l-yl)-2H-1
benzopyran;
10) (25, 3R, 4S) -6-amino-3,4-dihydro-3-hydr|oxy-2-
dimethoxymethyl-2-methyl-4-(2,3-dihydro-3,5,6-
trimethyl-2-cyanoimino-lH-benzimidazol-l-yl)-2H-1
benzopyran;
11) (2S, 3R, 4S)-6-nitro-3,4-dihydro-3-acet
dimethoxymethyl-2-methyl-4-(2,3-dihydro-2-cyanoim|ino-
IH-benzimidazol-l-yl)-2H-l-benzopyran;
12) (2S, 3R, 4S)-6-amino-3,4-dihydro-3-ace
dimethoxymethyl-2-methyl-4- (2, 3-dihydro-2-cyanoirr|ino-
IH-benzimidazol-l-yl)-2H-l-benzopyran;
13) (2S, 3R, 4S)-6-acetamino-3,4-dihi
hydroxy-2-dimethoxymethyl-2-methyl-4-(2, 3-dihydrc
cyanoimino-lH-benzimidazol-1-yl)-2H-l-benzopyran
14) (2S, 3R, 4S)-6-acetamino-3,4-dih
acetoxy-2-dimethoxymethyl-2-methyl-4-(2,3-dihydrc
cyanoimino-lH-benzimidazol-1-yl)-2H-1-benzopyran
15) (2S, 3R, 4S)-6-benzoylamino-3,4-dih;
hydroxy-2-dimethoxymethyl-2-methyl-4-(2,3-dihydr cyanoimino-lH-benzimidazol-1-yl)-2H-l-benzopyran
16) (23, 3R, 4S) -6- (trif luoroacetyl) ami:
dihydro-3-hydroxy-2-dimethoxymethyl-2-methyl-4-(i
oxy-2-
oxy-2-
dro-3-
-2-
dro-3-
-2-
dro-3-
-2-
0-3,4-
,3-
dihydro-2-cyanoimino-lH-benzimidazol-l-yl)-2H-lberizopyran;
17) (23, 3R, 4S) -6-methanesulfonylamincj>-3,4-
dihydro-3-hydroxy-2-dimethoxymethyl-2-methyl-4-(2,
dihydro-2-cyanoimino-lH-benzimidazol-l-yl)-2H-1-
benzopyran;
18) (2S, 3R, 4S)-6-nitro-3,4-dihydro-3-hydr dimethoxymethyl-2-methyl-4-(2,3-dihydro-2-oxo-lHbenzimidazol-
1-yl)-2H-l-benzopyran;
19) (25, 3R, 4S)-6-amino-3,4-dihydro-3-hydr dimethoxymethyl-2-methyl-4-(2,3-dihydro-2-oxo-lHbenzimida-
ol-l-yl)-2H-l-benzopyran;
20) (3R, 4S)-6-nitro-3,4-dihydro-3-hydroxV-2,2-
dimethyl-4- (2, 3-dihydro-2-oxo-lH-benzimidazol-l-y.L) -2H-
1-benzopyran;
21) (3R, 4S) -6-amino-3, 4-dihydro-3-hydrox|y-2, 2-
dimethyl-4-(2,3-dihydro-2-oxo-lH-benzimidazol-l-yL)-2H-
1-benzopyran;
22) (2S, 3R, 4S)-6-nitro-3,4-dihydro-3-hydrbxy-2-
dimethoxymethyl-2-methyl-4-(2,3-dihydro-2-thioxo-]LHbenzimidazol-
1-yl)-2H-l-benzopyran;
3-
23)(3R, 4S)-6-nitro-3,4-dihydro-3-hydrox
dimethyl-4- (2, 3-dihydro-2-thioxo-lH-benzimidazol-JL-yi;
2H-l-benzopyran;
24) (3R, 4S)-6-amino-3,4-dihydro-3-hydrox
y-2,2-
t-2,2-
dimethyl-4-(2,3-dihydro-2-thioxo-lH-benzimidazol- .-yl)-
2H-l-benzopyran;
25) (2S, 3R, 4S)-6-nitro-3,4-dihydro-3-hydrbxy-2-
diethoxymethyl-2-methyl-4-(2,3-dihydro-2-cyanoimino-lHbenzimidazol-
1-yl)-2H-l-benzopyran;
26) (2S, 3R, 4S)-6-nitro-3,4-dihydro-3-hydrbxy-2-
methoxymGthyl-2-methyl-4- (2, 3-dihydro-2-cyanoiminJD-lHbenzimidazol-
1-yl)-2H-l-benzopyran;
27) (2S, 3R, 4S)-6-amino-3,4-dihydro-3-hydroxy-2-
methoxymethyl-2-methyl-4-(2,3-dihydro-2-cyanoimino-lHbenzimidazol-
1-yl)-2H-l-benzopyran;
28) (2S, 3R, 4S)-6-nitro-3,4-dihydro-3-hydr|oxy-2-
methoxymethyl-2-methyl-4-(2,3-dihydro-2-cyanoimino-lH-
6-methylbenzimidazol-l-yl)-2H-l-benzopyran;
29) (2S, 3R, 4S)-6-nitro-3,4-dihydro-3-hyd4oxy-2-
([l,3]dioxan-2-yl)-2-methyl-4-(2,3-dihydro-2-
cyanoimino-lH-benzimidazol-l-yl)-2H-l-benzopyran;
30) (2S, 3R, 4S)-6-amino-3,4-dihydro-3-hydi|oxy-2-
([1,3]dioxan-2-yl)-2-methyl-4-(2,3-dihydro-2-
cyanoimino-lH-benzimidazol-1-yl)-2H-l-benzopyran;
31) (2S, 3R, 4S)-6-nitro-3,4-dihydro-3-hyd|oxy-2-
([l,3]dioxolan-2-yl)-2-methyl-4-(2,3-dihydro-2-
cyanoimino-lH-benzimidazol-1-yl)-2H-l-benzopyran;
32) (23, 3R, 4S)-6-nitro-3,4-dihydro-3-hyd2J-oxy-2-
[1,3]-5,5-dimethyldioxan-2-yl)-2-methyl-4-(2,3-
and
dihydro-2-cyanoimino-lH-benzimidazol-l-yl) "-2H-1-
benzopyran.
The compounds of of the p
invention are available in the form of pharmaceut
acceptable salts. And acid addition salts prepar
pharmaceutically acceptable free acids or metal s
re useful.
The acid salts of the compounds according •
present invention can be prepared in the cus
manner, for example by dissolving the compou
in excess aqueous free acid solution and
precipitating the salt using a water-miscible o
solvent, such as methanol, ethanol, acetor
acetonitrile. It is also possible to prepare th
salt by heating equivalent amounts of the compo'
and an free acid in water or alcohol
as glycol monomethyl ether, and then evaporatii
mixture to dryness or filtering off the precip
esent
:ally
sd by
Its a
o the
omary
d of
game
or
acid
nd of
such
a the
tated
salt with suction. Whether it is inorganic or orjganic,
a free acid can be used if it is pharmaceutlically
acceptable. Examples of the inorganic free
include hydrochloric acid, hydrobromic acid, su
acid, and phosphoric acid. Available organic free
acids are exemplified by citric acid, acetic
acid
Ifuric
acid,
lactic acid, tartaric acid, maleic acid, fumaric acid,
formic acid, propicnic acid, oxalic acid,
trifluoroacetic acid, benzoic acid, gluconic acid,
methanesulfonic acid, glycolic acid, succinic add, 4-
toluenesulfonic acid, galacturonic acid, embonic acid,
glutamic acid and aspartic acid.
Also, the compounds of may be in the
form of pharmaceutically acceptable alkali metal or
alkaline earth metal salts. The alkali metal or
alkaline earth metal salts of the compounds of 1> can be obtained, for example, by dissolviig the
compound of • in excess alkali metal or
alkaline earth metal hydroxide solution, filtering off
the undissolved materials and evaporating the fi.Iterate
to dryness. Sodium, potassium or calcium salts are
pharmaceutically suitable.
The present invention also provides proces
preparing benzopyran derivatives substitute
benzimidazole derivatives of .
Particularly, the present invention proprocess for preparing a compound of Formula (I)
is shown in . Reaction of a c
with a diamine compound (IV) in the presenc
proper metal salt gives a compound of Formula (V
compound
ses for
with
ides a
. which
of a
. Then,
cyclization of compound (IV) using an appropriate
reagent for introducing an X group affords a compound
(I'). Finally, a benzopyran compound substituted with
a benzimidazole (I) is prepared by changing
substituents R1, R2, R3, R4, R5 and R6. This is Refined
as 'preparation process 1' hereafter.
Scheme 1
(Figure Removed)

(Wherein, X, R1, R2, R3, R4, R5, R6 and *
defined in .)
are as
The present invention also provides another
process for preparing a compound of Formula (I) which
is shown in . cyclization of a diamine
compound (IV) using proper reagent gives a compound of
Formula (VI). Then, epoxide ring opening of compound
(111) is accomplished by reaction with a compound (VI) in
the presence of a proper base, giving a compound of
Formula (!')• Finally, a benzopyran con pound
substituted with a benzimidazole (I) is prepared by
introduing substituents R1, R2, R3, R4, R5 and R6. This
is defined as 'preparation process 2' hereafter.

(Figure Removed)

(Wherein, X, R1, R2, R3, R4, R5, R6 and *
defined in .)
are as
In the present invention, a compound of formula
1> can be prepared in the form of an ind:ijvidual
diastereomer from the corresponding diastereoner of
starting material. Each diastereomer can a!.so be
obtained by separating the diastereomeric mixtbre of
compound (I) prepared from a diasteremeric mixture of
starting material. The separation of diastereomers can
be carried out by column chromatography or
reerystallization.
The preparation processes for benzjopyran
derivatives substituted with a benzimidazole derivative
represented in of the present invention are
illustrated in more detail hereafter.
I. Preparation of starting material
(1) Preparation of epoxide compound (III)
Epoxide compound (III ) used as a starting material
in can be prepared by processes described in
Korean Patent No. 2000-60647 and U.S. Patent 6,3J23,238.
As shown in the , each diastereomer (III
i) , (Ilia), (Ills) and (III 4) of a compound (III) I can be
possibly prepared from olefin compounds (Vlli) and (Vila)
by employing Mn (III) Salen epoxidation patalyst
described in the above patents. !

(Figure Removed)
(Wherein, R1 and R2 are as defined in II. Preparation process 1
The preparation process for a compound of Formula
(I) represented in comprises the following
steps:
1) preparing compound (V ) by reaction of
epoxide compound (111) with diamine
compound (IV) in the presence of a proper
metal salt in proper solvent;
2) preparing compound (I') by cyclization of
diamine compound (V ) using an appropriate
reagent for introducing X group; and
3) preparing compound (I) by changing
substituents of the compound (I')-
In the step 1) is a reaction of epoxide cojnpound
[III] with diamine compound (IV) in the presence! of a
proper metal salt in proper solvent.
As a metal salt, Mg(C104}2, CoCl2, LiClO4, NkC104,
CaCl2, ZnCl2, LiBF4 or Zn(Tf)2 can be used. As a scjlvent,
I
acetonitrile, tetrahydrofuran or dimethylformamidja can
be used and acetonitrile is preferred. Reaction
temperature ranges from room temperature to the boiling
point of the solvent.
In case that an individual stereoisomeij
epoxide compound (III) is used as starting material,!
of
the
stereroisomer with a stereochemistry corresponding to
the stereoisomer used as starting material wil4 be
i
obtained. As shown in the below , compojunds
(V i) , (V2), (V3) and (V4) are prepared from jaach
epoxide compound (Illi), (III2), (III3) and (III4). i
I

(Figure Removed)

As shown in , in the above step 2), the
compound of in which X is 0, S or NCN can
be prepared from a compound (V) by performing a
cyclization using an appropriate reagent.

(Figure Removed)
(Wherein, R1, R2, R5, R6 and * are as defined in
.)
The compound of formula (Ii' ) in which X is O can
be prepared by using carbonyl transfer reagent derived
from phosgene such as phosgene, urea, dimethyl
carbonate, carbonyldiimidazole, triphosgene, 1,1'-
carbonyl-di-1,2,4-triazole, 1-disuccinimidyl carbonate,
di-2-pyridyl carbonate, etc.
The compound of formula (3V ) in which X is S can
be prepared by using thiocarbonyl transfer reagent
derived from thiophosgene such as thiophosgene,
thiourea, 1,1-thiocarbonyldiimidazole, 1/1'-
thiocarbonyldi-1,2,4-triazole, di-2-pyridyl
thiocarbonate, 1,1'-thiocarbonyl-2,2'-pyridone, etc.
The compound of formula (I3' ) / in which X is NCN
can be prepared by using diphenyl cyanocarbonimidate or
N-cyanodithioiminocarbonate.
In the above step 3) , a compound (I) of 1> is prepared by changing substituents R1, R2, R3, R4,
R5 and R6 by alkylation, acylation, reduction, or
substitution, etc.
For example, as shown in , if R1 of a
compound (I) is amino group, the compound can be
prepared by reducing nitro group, for which
hydrogenation is performed using a metal catalyst such
as platinum, palladium on carbon (Pd/C) or Raney-nickel
in proper solvent. Alternatively, a nitro group can be
reduced by a reducing agent like NaBH4 in the presence
of CuS04, Cu(OAc)2, CoCl2, SnCl2 or NiCl2. In this
reaction, preferable solvent is a mixture of water and
methanol and reaction temperature ranges from room
temperature to the boiling point of the solvent.

(Figure Removed)
(Wherein, R2, R3, R4, R5, R6 and * are as defined
in .)
III. Preparation process 2
Another process to prepare a compound (I) of
is illustrated in . In step 1) of
, as shown in , Cyclization of
diamine compound (IV) gives compounds (Vli, VI2, and VI3) ,
using such reagents as in step 2) of the preparation
process 1.

(Figure Removed)

In step 2), a compound (I') is prepared by
epoxide ring opening, in which a compound (VI) is
reacted with epoxide compound (III) in the presence of
base. Both inorganic base such as sodium hydride,
potassium t-butoxide, sodium methoxide, etc. and
organic base such as 1,8-diazabicyclo[5.4.0]undec-7-ene
(DBU), etc, can be used.
In step 3), a compound (I) is prepared by
changing substituents as described in the preparation
process 1.
The present invention further provides
pharmaceutical composition for cardioprotection
containing benzopyran derivatives substituted with a
benzimidazole derivative, represented in ,
or pharmaceutically acceptable salts thereof as an
effective ingredient.
When tested in ischemic heart models of
Langendorff using isolated rat hearts, compounds of the
present invention significantly prolong the time to
contracture (TTC) , an index of heart protection, and
improve recovery of the cardiac function (left
ventricular developed pressure x heart rate, LVDP x HR)
after reperfusion, but reduce release of lactate
dehydrogenase (LDH), an index for cell damage, which
are similar or superior to cardioprotecting activity of
BMS-180448, a control. In ischemic myocardium models
using anesthetized rat, compounds of the present
invention also show similar antiischemic activity to
BMS-180448. In the tests with blood vessels isolated
from a white rat, the compounds of the present
invention shows better cardioselective antiischemic
activity than BMS-180448 owing to their minor
vasorelaxant activity.
In conclusion, the compounds of the present
invention do not drop blood pressure owing to their
minor vasorelaxant activity but have excellent
antiischemic activity. Therefore, the compounds of the
invention can be effectively used not only for the
protection of heart but also for the prevention or the
treatment of ischemic heart diseases such as myocardial
infarction and unstable angina pectoris and ischemiareperfusion
related diseases caused by thrombolytics or
reperfusxon therapy like PTCA (percutaneous
transluminal coronary angioplasty) and CABG (coronary
artery bypass graft), decrease of myocardial
contractility, myocardial injury, change of energy
metabolism and decline of cognitive capability. In
addition, the compounds of the present invention can be
used as a protective agent against brain injury, a
protective agent for retinal cells or organs for longterm
storage such as heart, kidney, liver and tissues,
or a treating agent for isr.hemia-reperfu'sion related
diseases.
EXAMPLES
In the following the invention is described in
more detail with reference to examples. These examples
are intended for illustration only and are not to be
construed as any limitation.
In the present invention, infrared spectroscopy,
nuclear magnetic resonance spectroscopy, mass
spectroscopy, liquid chromatography, x-ray
crystallography, polarimetry were used along with the
comparison of estimated results of elemental analysis
of the representative compounds with analyzed results
of them in order to confirm their molecular structures.
Example 1: Preparation of (25, 3R, 45)-6-nitro-3,4-
dihydro-3-hydroxy-2-dimethoxymethyl-2-methyl-4-(2,3-
dihydro-2-cyanoimino-lH-benzimidazol-l-yl)-2H-1-
benzopyran
Preparation of (2S, 3R, 4S)-6-nitro-3,4-
dihydro-3-hydroxy-2-dimethoxymethyl-2-methyl-4-[(2-
aminophenyl)amino]-2H-l-benzopyran
950 mg (3.38 mmol) of epoxide compound (2S, 3R,
4R)-6-nitro-3,4-dihydro-3,4-epoxy-2-dimethoxymethyl-2-
methyl-2H-l-benzopyran and 370 ing (3.38 mmol) of 1,2-
phenylinediamine were dissolved in 3 M of acetonitrile
(CH3CN), then 754 mg (3.38 mmol) of magnesium
perchlorate [Mg(C104)2] was added thereto. The reaction
is stirred at room temperature for 2 hours, 10 M£ of
saturated NaHC03 solution was added, and aqueous layer
was extracted with 30 M of ethyl acetate. Combined
organic layer was dried over anhydrous MgS04, filtered
and concentrated under reduced pressure. The residue
was purified by column chromatography (hexane: ethyl
acetate = 1:1), to give 670 mg (yield: 51%) of the
target compound.
ltt NMR (200 MHz, CDC13) 6l.48(s, 3H),3.53(s, 3H) ,
3.55(s, 3H) , 4.08(d, 1H) , 4.47(s, 1H) , 4.62(d,
6.79(m, 5H) , 8.06(dd, 1H) , 8.32(d,
Preparation of (2S, 3R, 4S) -6-nitro-3, 4-
dihydro-3-hydroxy-2-dimethoxymethyl-2-methyl-4- (2,3-
dihydro-2-cyanoimino-lH-benzimidazol-l-yl)-2H-lbenzopyran
590 mg (1.52 mmol) of the compound obtained in the
above step 1 was dissolved in 6 M of i-PrOH, then 360
mg (1.52 mmol) of diphenyl cyanocarbonimidate and 424 \lt
(3.04 mmol) of triethylamine were added thereto. The
reaction was stirred and refluxed for 12 hours, 30 M£ of
saturated NaHC03 solution was added, aqueous layer was
extracted with 40 M of ethyl acetate. Combined organic
layer was washed with brine and dried over anhydrous
MgS04, and concentrated under reduced pressure. The
residue was purified by silica gel column
chromatography (hexane-.ethyl acetate = 1:1), to give
420 mg (yield: 64%) of the target compound.
1H NMR (200 MHz, CDC13) 6l.66(s, 3H) , 3.50(s, 3H) ,
3.55(s, 3H), 4.21(d, 1H) , 4.56(s, 1H) , 6.11(d,
6.25(d, 1H), 6.90(m, 1H) , 7.11(m, 2H) , 7.28(m,
7.76(d, 1H) , 8.11(dd,
Example 2: Preparation of (2R, 3R, 4S) -6-nitro-3, 4-
dihydro-3-hydroxy-2-dimethoxymethyl-2-methyl-4- (2,3-
dihydro-2-cyanoimino-lH-benzimidazol-l-yl)-2H-lbenzopyran
Preparation of (2R, 3R, 4S) -6-nitro-3, 4-
dihydro-3-hydroxy-2-dimethoxymethyl-2-methyl-4- [ (2-
aminophenyl) amino] -2H-l-benzopyran
540 mg (1.92 mmol) of epoxide compound (2R, 3R,
4R) -6-nitro-3, 4-dihydro-3, 4-epoxy-2-dimethoxymethyl-2-
methyl-2H-l-benzopyran and 208 mg (1.92 mmol) of 1,2-
phenylinediamine were reacted in analogy to the
procedure described in step 1 of the example 1, to give
404 mg (yield: 54%) of the target compound.
XH NMR (200 MHz, CDC13) 5l.43(s, 3H) , 3.50(s, 3H) ,
3.55(s, 3H), 4.14(d, 1H) , 4.45(s, 1H) , 4.49(d,
6.75(m, 5H), 8.09(dd, 1H), 8.32(d,
Preparation of (2R, 3R, 4S)-6-nitro-3,4-
dihydro-3-hydroxy-2-dimethoxymethyl-2-methyl-4-(2, 3-
dihydro-2-cyanoimino-lH-benzimidazol-l-yl)-2H-lbenzopyran
Reaction of 404 mg (1.04 mmol) of the compound
prepared in the above step 1 was performed in analogy
to the procedure described in the step 2 of the example
1, to give 306 mg (yield: 67%) of the target compound.
1H NMR (200 MHz, CDC13) 5l.49(s, 3H), 3.62(s, 3H),
3.64(s, 3H), 4.51(s, 1H) , 4.78(d, 1H) , 5.90(d, 1H) ,
6.40(d, 1H), 6. 9-7.4(m, 4H), 7.79(d, 1H), 8.14(dd, 1H)
Example 3; Preparation of (2S, 3S, 4R)-6-nitro-3,4-
dihydro-3-hydroxy-2-dimethoxymethyl-2-methyl-4-(2,3-
dihydro-2-cyanoimino-lH-benzimidazol-l-yl)-2H-1-
benzopyran
Preparation of (2S, 3S, 4R)-6-nitro-3,4-
dihydro-3-hydroxy-2-dimethoxymethyl-2-methyl-4-[(2-
aminophenyl)amino]-2H-l-benzopyran
Reaction of 2 g (7.11 mmol) of epoxide compound
(2S, 3S, 4S)-6-nitro-3,4-dihydro-3,4-epoxy-2-
dimethoxymethyl-2-methyl-2H-l-benzopyran with 1.15 g
(10.7 ramol) of 1,2-phenylinediamine were performed in
analogy to the procedure described in step 1 of the
example 1, to give 2.08 g (yield: 75%) of the target
compound.
XH NMR (200 MHz, CDC13) 6l.36(s, 3H), 3.58(s, 3H),
3.59(s, 3H), 4.23(d, 1H) , 4.41(s, 1H) , 4.51(d, 1H) ,
6.72-6.78(m, 4H) , 6.90(d, 1H) , 8.03(dd, 1H) , 8.34(d,
1H)
Mass : 389, 296, 119, 108, 75
Preparation of (2S, 3S, 4R)-6-nitro-3, 4-
dihydro-3-hydroxy-2-dimethoxymethyl-2-methyl-4-(2,3-
dihydro-2-cyanoimino-lH-benzimidazol-l-yl)-2H-1-
benzopyran
Reaction of 1.51 g (3.88 mmol) of the compound
prepared in the above step 1 was performed in analogy
to the procedure described in step 2 of the example 1,
to give 506 mg (yield: 30%) of the target compound.
1H NMR (200 MHz, CDC13) 6l.49(s, 3H), 3.62(s, 3H),
3.64(s, 3H), 4.50(s, 1H) , 4.77(d, 1H) , 5.90(d, 1H) ,
6.37(d, 1H), 6.92(t, 1H), 7.06-7.14(m, 2H) , 7.29(d, 1H),
7.76(d, 1H), 8.12{dd, 1H)
Mass : 439, 250, 190, 158, 75
Example 4; Preparation of (2R, 3S, 4S)-6-nitro-3,4-
dihydro-3-hydroxy-2-dimethoxymethyl-2-methyl-4-(2,3-
dihydro-2-cyanoimino-lH-benzimidazol-l-yl)-2H-1-
benzopyran
Preparation of (2R, 3S, 4S)-6-nitro-3,4-
dihydro-3-hydroxy-2-dimethoxymethyl-2-methyl-4-[(2-
aminophenyl)amino]-2H-l-benzopyran
Reaction of 1.50 g (5.33 mmol) of epoxide
compound (2R, 3R, 4R)-6-nitro-3,4-dihydro-3,4-epoxy-2-
dimethoxymethyl-2-methyl-2H-l-benzopyran with 692 mg
(6.40 mmol) of 1,2-phenylinediamine was performed in
analogy to the procedure described in step 1 of the
example 1, to give 1.74 g (yield: 84%) of the target
compound.
XH NMR (200 MHz, CDC13) 6l.48(s, 3H), 3.53(s, 3H),
3.55(s, 3H), 3.88(br-s, OH), 4.09(d, 1H), 4.48(s, 1H) ,
4.64(br-s, 1H), 6.71-6.97(m, 5H), 8.06(dd, 1H), 8.32(d,
1H)
Mass : 388, 295, 119, 108, 75
Preparation of (2R, 3S, 4S)-6-nitro-3,4-
dihydro-3-hydroxy-2-dimethoxymethyl-2-methyl-4-(2,3-
dihydro-2-cyanoimino-lH-benzimidazol-l-yl)-2H-l-
benzopyran
Reaction of 1.74 g (4.47 mmol) of the compound
prepared in the above step 1 was performed in analogy
to the procedure described in step 2 of the example 1,
to give 1.12 g (yield: 57%) of the target compound.
1H NMR (200 MHz, CDC13) 6l.66(s, 3H), 3.50(s, 3H),
3.56(s, 3H) , 4.24(d, 1H) , 4.57(s, 1H) , 6.14(d, 1H) ,
6.27(d, 1H), 6.90(t, 1H), 7.05-7.13(m, 2H), 7.31(d, 1H) ,
7.76(d, 1H) , 8.14(dd, 1H)
Mass : 439, 250, 190, 75
Example 5: Preparation of (2S, 3R, 4S)-6-amino-3,4-
dihydro-3-hydroxy-2-dimethoxymethyl-2-methyl-4-(2,3-
dihydro-2-cyanoimino-lH-benzimidazol-l-yl)-2H-1-
benzopyran
150 mg (0.34 mmol) of the compound obtained in the
example 1 was dissolved in 3 M of methanol, to which
20 mg of 10% Pd/C was added. The reaction was stirred
for 5 hours at room temperature under 3 atm of hydrogen
gas. The reaction solution was filtered with celite
pad to eliminate solid substances, the filterate
concentrated under reduced pressure. The residue was
purified by silica gel column chromatography

(hexane:ethyl acetate = 2:1), to give 120 mg (yield:
80%) of the target compound.
aH NMR (200 MHz, CDC13) 5l.55(s, 3H) , 3.49(br-s,
2H, NH2), 3.53(sf 3H), 3.57(s, 3H), 4.23(d, 1H), 4.59(s,
, 5.81(d, 1H), 6.18(d, 1H) , 6.48(d, 1H) , 6.60(dd,
, 6.78(d, 1H), 6.91(dd, 1H) , 7.07(dd, 1H) , 7.24(d,
1H)
Example 6: Preparation of (2S, 3R, 4S)-6-nitro-3,4-
dihydro-3-hydroxy-2-dimethoxymethyl-2-inethyl-4-(2,3-
dihydro-3-methyl-2-cyanoimino-lH-benzimidazol-l-yl)-2H-
1-benzopyran
400 mg (0.91 mmol) of the compound obtained in
example 1 was dissolved in 5 M£ of DMF, then 250 mg
(1.82 mmol) of K2C03 and 170 mg (1.18 mmol) of CH3I were
added thereto. The reaction was stirred at room
temperature for 12 hours, 30 M of saturated NaHC03
solution was added, and aqueous layer was extracted
with 50 M£ of ethyl acetate. Organic layer was washed
with brine and dried over anhydrous MgS04, concentrated
under reduced pressure. The residue was purified by
silica gel column chromatography (hexane:ethyl acetate
= 1:1), to give 390 mg (yield: 95%) of the target
compound.
XH NMR (200 MHz, CDC13) 61.65 (s, 3H) , 3.50 (s,
3H), 3.55 (s, 3H), 4.56 (s, 1H), 6.23 (d, 1H) , 6.52 (d,
1H), 6.92 (m, 2H), 6.96 (d, 2H), 7.06 (d, 2H), 7.82 (d,
1H), 8.15 (dd, 1H)
Example 7: Preparation of (2S, 3R, 4S)-6-nitro-3,4-
dihydro-3-hydroxy-2-dimethoxymethyl-2-methyl-4-(2,3-
dihydro-5,6-dimethyl-2-cyanoimino-lH-benzimidazol-lyl)-
2H-l-benzopyran
Preparation of (2S, 3R, 4S)-6-nitro-3,4-
dihydro-3-hydroxy-2-dimethoxymethyl-2-methyl-4-[(2-
amino-4, 5-dimethylphenyl).amino] -2H-l-benzopyran
Reaction of 480 ing (1.71 nunol) of epoxide compound
(2S, 3R, 4R)-6-nitro-3,4-dihydro-3,4-epoxy-2-
dimethoxymethyl-2-methyl-2H-l-benzopyran with 232 mg
(1.71 mmol) of 4,5-dimethyl-l,2-phenylinediamine was
performed in analogy to the procedure described in step
1 of the example 1, to give 307 mg (yield: 43%) of the
target compound.
*H NMR (200 MHz, CDC13) 6l.47(s, 3H), 2.16(s, 3H),
2.19(s, 3H) , 3.53(s, 3H) , 3.55(s, 3H) , 4.08(d,
4.47{s, 1H), 4.56(d, 1H) , 6.61(s, 1H) , 6.65(s,
6.93(d, 1H), 8.07(dd, 1H) , 8.34(d,
Preparation of (2S, 3R, 4S) -6-nitro-3, 4-
dihydro-3-hydroxy-2-dimethoxymethyl-2-methyl-4- (2,3-
dihydro-5, 6-dimethyl-2-cyanoimino-lH-benzimidazol-lyl)
-2H-l-benzopyran
Reaction of 110 mg ( 0 . 2 6 mmol) of the compound prepared
in the above step 1 was performed in analogy tc the
procedure described in step 2 of the example 1, tc give
74 mg (yield: 58%) of the target compound.
XH NMR (200 MHz, CDC13) 6 l . 6 7 ( s , 3H) , 2 . 0 7 ( s , 3H) ,
2 . 2 0 ( s , 3H), 3.51(3, 3H), 3.55(5, 3H) , 4.20(d, 1H) ,
4 . 5 5 ( s , 1H), 6.01(s, 1H), 6.06(d, 1H) , 7.04(d, 1H) ,
7 . 0 7 ( 3 , 1H) , 7 . 7 4 ( d , 1H) , 8.13(dd, 1H)
Example 8: Preparation of (2S, 3R, 4S) -6-amino-3, 4-
dihydro-3-hydroxy-2-dimethoxymethyl-2-methyl-4- ( 2 , 3-
dihydro-5, 6-dimethyl-2-cyanoimino-lH-benzimidazol-lyl)
-2H-l-benzopyran
Reaction of 210 mg (0.43 mmol) of a nitro compound
prepared in example 7 was performed in analogy to the
procedure described in the example 5, to give 177 mg
(yield: 91%) of the target compound.
*H NMR (200 MHz, CDC13) 6l.54(s, 3H) , 2.15(s, 3H) ,
2.17(s, 3H) , 3.52(s, 3H) , 3.54(s, 3H) , 4.25(d,
4.66(s, 1H) , 6.14(d, 1H) , 6.17(d, 1H) , 6.39(s,
6.56(dd, 1H) , 6.74(d, 1H) , 7.32(s,
Example 9: Preparation of (2S, 3R, 4S) -6-nitro-3, 4-
dihydro-3-hydroxy-2-dimethoxymethyl-2-methyl-4- (2, 3-
dihydro-3, 5, 6-trimethyl-2-cyanoimino-lH-benzimidazol-lyl)
-2H-l-benzopyran
Reaction of 180 mg (0.37 mmol) of the compound prepared
in example 7 was performed in analogy to the procedure
described in the example 6, to give 144 mg (yield: 81%)
of the target compound.
1E NMR (200 MHz, CDC13) 6l.65(s, 3H) , 2.07(s, 3H) ,
2.26(8, 3H), 3.51(s, 3H) , 3.54(s, 3H) , 3.87(s, 3H) ,
4.55(s, 1H) , 5.98(s, 1H) , 6.44(d, 1H) , 6.94(s,
7.05(d, 1H) , 7.80(d, 1H) , 8.17(dd,
Example 10: Preparation of (2S, 3R, 4S) -6-amino-3, 4-
dihydro-3-hydroxy-2-dimethoxymethyl-2-methyl-4-(2,3-
dihydro-3, 5, 6-trimethyl-2-cyanoimino-lH-benzimidazol-lyl)
-2H-l-benzopyran
Reaction of 70 mg (0.15 mmol) of a nitro compound
prepared in example 9 was performed in analogy to the
procedure described in the example 5, to give 58 mg
(yield: 89%) of the target compound.
ltt NMR (200 MHz, CDC13) 6l.56(s, 3H) , 2.12(s, 3H) ,
2.28(s, 3H) , 3.53(s, 3H) , 3.59(3, 3H) , 3.80(s, 3H) ,
4.17(t, 1H) , 4.62(s, 1H) , 6.24(s, 1H) , 6.26(d,
6.64(dd, 1H), 6.78(d, 1H) , 6.91(s,
Example 11: Preparation of (2S, 3R, 4S) -6-nitro-3, 4-
dihydro-3-acetoxy-2-dimethoxymethyl-2-methyl-4- (2,3-
dihydro-2-cyanoimino-lH-benzimidazol-l-yl)-2H-lbenzopyran
388 mg (0.88 mmol) of the compound obtained in the
example 1 was dissolved in 4 M of methylenechloride,
then 83 \i£ (0.88 mmol) of acetic anhydride, 0.18 M0
(1.32 mmol) of triethylamine and 32 mg (0.26 mmol) of
4-dimethylaminopyridine were added thereto. The
reaction was stirred at room temperature for 2 hours,
30 M of saturated NaHC03 solution was added, aqueous
layer was extracted with 60 M£ of methylenechloride.
Organic layer was washed with brine and dried over
anhydrous MgSO4, concentrated under reduced pressure.
The residue was purified by silica gel column
chromatography (hexane:ethyl acetate = 2:1), to give
376 mg (yield: 89%) of the target compound.
XH NMR (200 MHz, CDC13) 6l.49(s, 3H) , 2.10(s, 3H),
3.52(s, 3H), 3.55(s, 3H) , 4.63(s, 1H) , 5.62(d,
6.24(d, 1H), 6.31(d, 1H), 6.91-7.24(m, 3H), 7.33(d,
7.70(d, 1H), 8.13(dd, 1H)
Mass : 481(M+)
Example 12: Preparation of (2S, 3R, 4S)-6-amino-3,4-
dihydro-3-acetoxy-2-dimethoxymethyl-2-methyl-4-(2,3-
dihydro-2-cyanoimino-lH-benzimidazol-l-yl) -2H-1-
benzopyran
196 mg (0.41 mmol) of a nitro compound obtained in
the example 11 was dissolved in 10 M of methanol, to
which 98 mg of Raney-Ni was added. The reaction was
stirred for 15 hours at room temperature under 3 atm of
hydrogen gas. The reaction solution was filtered to
eliminate Ni, followed by concentration under reduced
pressure. The residue was purified by silica gel
column chromatography (hexane:ethyl acetate = 1:2), to
give 70 mg (yield: 38%) of the target compound.
:H NMR (200 MHz, CDC13) 6l.40(s, 3H), 2.06(s, 3H) ,
3.35(br-s, 2H, -NH2), 3.47(s, 3H) 3.55(s, 3H), 4.56(s,
1H), 5.63(d, 1H), 6.05(d, 1H), 6.12(d, 1H), 6.53(d, 1H),
6.59(dd, 1H), 6.81(d, 1H), 6.96(dd, 1H), 7.11(dd,
7.29(d,
Example 13: Preparation of (2S, 3R, 4S)-6-acetamino-
3,4-dihydro-3-hydroxy-2-dimethoxymethyl-2-methyl-4-
(2,3-dihydro-2-cyanoimino-lH-benzimidazol-l-yl)-2H-1-
benzopyran
Reaction of 100 mg (0.24 mmol) of the compound
prepared in example 5 was performed in analogy to the
procedure described in the example 11, to give 62 mg
(yield: 57%) of the target compound.
XH NMR (200 MHz, CDC13) 5l.49(s, 3H) , 1.93(s, 3H) ,
3.44(s, 3H), 3.46(s, 3H) , 4.13(dd, 1H) , 4.52(s, 1H) ,
5.81(d, 1H), 6.34(d, 1H), 6.71(d, 1H), 6.80-6.87(m, 2H),
7.01(dd, 1H), 7.20(d, 1H), 7.68(dd, 1H), 8.59(s, 1H, -
NH), 12.26(3, 1H, -NH)
Example 14: Preparation of (2S, 3R, 4S)-6-acetamino-
3,4-dihydro-3-acetoxy-2-dimethoxymethyl-2-methyl-4-
(2,3-dihydro-2-cyanoimino-lH-benzimidazol-l-yl)-2H-1-
benzopyran
88 mg (0.21 mmol) of the compound obtained in
example 5 was dissolved in methylenechloride, then 61
\i£ (0.64 mmol) of acetic anhydride, 120 \& (0.86 mmol)
of triethylamine and 8 mg (0.06 mmol) of 4-
dimethylaminopyridine were added thereto. The reaction
was stirred at room temperature for 12 hours, saturated
NaHC03 solution was added, aqueous layer was extracted
with methylenechloride. Organic layer was washed with
brine and dried over anhydrous MgS04, concentrated
under reduced pressure. The residue was purified by
silica gel column chromatography (hexane:ethyl acetate
= 1:2), to give 35 mg (yield: 34%) of the target
compound.
1H NMR (200 MHz, CDC13) 5l.43(s, 3H) , 2.02(s, 3H) ,
2.07(S, 3H), 3.48(s, 3H) , 3.54(s, 3H) , 4.59(s,
5.63(d, 1H), 6.14(d, 1H), 6.57(d, 1H) , 6.67(d,
6.94-6.98(m, 2H), 7.08-7.15(m, 2H), 7.62(dd,
11.85(br-s,
Example 15: Preparation of (2S, 3R, 4S)-6-benzoylamino-
3,4-dihydro-3-hydroxy-2-dimethoxymethyl-2-methyl-4-
(2,3-dihydro-2-cyanoimino-lH-benzimidazol-l-yl)-2H-1-
benzopyran
100 ing (0.24 mmol) of the compound obtained in
example 5 was dissolved in 1 M of THF, then 28 \i£ (0.24
mmol) of benzoyl chloride and 51 \i& (0.37 mmol) of
triethylamine were added thereto. The reaction was
stirred at room temperature for 2 hours, 10 M of
saturated NaHC03 solution was added, and aqueous layer
was extracted with 20 M of ethyl acetate. Organic
layer was dried over anhydrous MgS04, and concentrated
under reduced pressure. The residue was purified by
silica gel column chromatography (hexane:ethyl acetate
= 1:2), to give 68 mg (yield: 54%) of the target
compound.
:H NMR (200 MHz, CDC13) 5l.60(s, 3H) , 3.55(s, 6H) ,
4-25(dd, 1H), 4.61(s, 1H), 5.95(d, 1H) , 6.44(d, 1H) ,
6.87-6.94(m, 2H), 7.00(d, 1H), 7.08(dd, 1H), 7.37-
7.48(111, 3H), 7.77(m, 3H) , 7.85(dd, 1H)
Example 16: Preparation of (2S, 3R, 4S)-6
(trifluqroacetyl)amino-3,4-dihydro-3-hydroxy-2-
dimethoxymethyl-2-methyl-4-(2.3-dihvdro-2-cyanoiminolH-
benzimidazol-l-yl)-2H-l-benzopyran
Reaction of 120 mg (0.29 mmol) of the compound
prepared in the example 5 with 41 \l£ (0.29 mmol) of
trifluoroacetic anhydride was performed in analogy to
the procedure described example 11 to give 30 mg
(yield: 21%) of the target compound.
1E NMR (200 MHz, CDC13) 5l.61(s, 3H) , 3.53{s,- 3H) ,
3.55(s, 3H) , 4.23(m, 1H) , 4.57(s, 1H) , 5.98(d, 1H) ,
6.39(d, 1H) , 6.86-7.14(m, 4H) , 7.73(dd, 1H) , 8.06(s,
1H)
Example 17: Preparation of (2S, 3R, 4S)-6-
methanesulfonylamino-3,4-dihydro-3-hydroxy-2-
dimethoxymethyl-2-methyl-4-(2,3-dihydro-2-cyanoiminolH-
benzimidazol-1-yl)-2H-l-benzopyran
120 mg (0.29 mmol) of the compound obtained in
example 5 was dissolved in 2 M of methylenechloride,
then 23 ^ (0.29 mmol) of methanesulfonyl chloride and
76 v& (0.44 mmol) of diisopropylethylamine were added
thereto. The reaction was stirred at room temperature
for 14 hours, saturated NaHC03 solution was added, and
aqueous layer was extracted with 30 W of
methylenechloride. Organic layer was dried over
anhydrous MgS04, concentrated under reduced pressure.
The residue was purified by silica gel column
chromatography (hexane : ethyl acetate = 1:1), to give 35
mg (yield: 26%) of the target compound.
1E NMR (200 MHz, CDC13) 5l.60(s, 3H) , 2.62(3, 3H) ,
3.55(s, 6H) , 4.20(dd, 1H) , 4.59(s, 1H) , 5.96{d, 1H) ,
6.33(d, 1H) , 6.67(d, 1H) , 6. 81-6. 98 (m, 3H) , 7.08(dd,
1H) , 7.31(m, 1H)
Example 18: Preparation of (2S, 3R, 45) -6-nitro-3, 4-
dihydro-3-hydroxy-2-dimethoxymethyl-2-methyl-4- (2,3-
dihydro-2-oxo-lH-benzimidazol-l-yl) -2H-l-benzopyran
391 mg (1.0 mmol) of the compound obtained in the
step 1 of the example 1 was dissolved in 4 M of
methylenechloride, then 262 mg (1.21 mmol) of di-2-
pyridyl carbonate and 12 mg (0.10 mmol) of 4-
dimethylaminopyridine were added thereto. The reaction
was stirred at room temperature for 1 hour, solvent was
evaporated under reduced pressure. The residue was
purified by silica gel column chromatography, to give
193 mg (yield: 47%) of the target compound.
*H NMR (200 MHz, CDC13) 5l.26(s, 3H), 3.47(s, 3H),
3.53(s, 3H), 4.23(br, 2H) , 4.57(s, 1H) , 5.96(d,
6.01(d, 1H), 6.80(t, 1H), 6.99-7.26(m, 4H), 7.89(s,
8.10(dd, 1H), 8.91(s, 1H)
Mass : 415, 324, 206, 190
Example 19: Preparation of (2S, 3R, 43)-6-amino-3,4-
dihydro-3-hydroxy-2-dimethoxymethyl-2-methyl-4-(2,3-
dihydro-2-oxo-lH-benzimidazol-l-yl)-2H-l-benzopyran
Reaction of 100 mg (0.24 mmol) of the compound
prepared in example 18 was performed in analogy to the
procedure described in the example 5, to give 58 mg
(yield: 63%) of the target compound.
1H NMR (200 MHz, CDC13) 6l.54(s, 3H) , 3.36(br,
NH2), 3.52(s, 6H) , 4.19(t, 1H) , 4.61(s, 1H) , 5.67(d,
1H) , 6.33(m, 2H), 6.57(m, 1H), 6.77(m, 2H), 6.95(m, 2H),
9.73(s, NH)
Mass : 385(M+), 292, 176, 160
Example 20: Preparation of (3R, 4S) -6-nitro-3, 4-
dihydro-3-hydroxy-2,2-dimethyl-4-(2,3-dihydro-2-oxo-lHbenzimidazol-
1-yl) -2H-l-benzopyran
Preparation of (3R, 4S) -6-nitro-3, 4-dihydro-3-
hydroxy-2,2-dimethyl-4- [ (2-aminophenyl) amino] -2H-1-
benzopyran
Reaction of 1.0 g (4.53 mmol) of epoxide compound
(3R, 4R) -6-nitro-3, 4-dihydro-3, 4-epoxy-2, 2-dimethyl-2H-
1-benzopyran with 0.49 g (4.53 mmol) of 1,2-
phenylinediamine was performed in analogy to the
procedure described in the step 1 of the example 1, to
give 0.51 g (yield: 32%) of the target compound.
XH NMR (200 MHz, CDC13) 6l.34(s, 3H) , 1.53(s, 3H) ,
3.75(d, 1H), 4.51(d, 1H), 6.70(m, 5H) , 8.01(dd,
8.28(s,
Preparation of (3R, 4S) -6-nitro-3, 4-dihydro-3-
hydroxy-2 , 2-dimethyl-4- (2, 3-dihydro-2-oxo-lHbenzimidazol-
l-yl)-2H-l-benzopyran
Reaction of 504 mg (1.80 mmol) of the compound
prepared in the above step 1 was performed in analogy
to the procedure described in the example 18, to give
410 mg (yield: 65%) of the target compound.
1H NMR (200 MHz, CDC13) 6l.42(s,3H), 1.62(s,3H),
3.59(br-s,lH-OH) , 4.18(m,1H), 5.67(d,lH), 6.15(d,lH),
6.77(t,lH), 6.93(m,3H), 7.82(s,lH), 8.10(dd,lH), (brs,
lH)
Example 21: Preparation of (3R, 4S)-6-amino-3,4-
dihydro-3-hydroxy-2,2-dimethyl-4-(2,3-dihydro-2-oxo-lHbenzimidazol-
l-yl)-2H-l-benzopyran
Reaction of 130 ing (0.37 mmol) of the compound
prepared in example 20 was performed in analogy to the
procedure described in the example 5, to give 110 mg
(yield: 92%) of the target compound.
1H NMR (200 MHz, CDC13) 6l.37(s, 3H) , 1.50(s, 3H) ,
3.37(br-s, 2H, -NH2), 4.07(br-s, 1H, -OH), 5.39(d, 1H),
6.31(dd, 1H) , 6.62(dd, 1H) , 6.74(d, 1H) , 6.91(m, 2H) ,
7.20(d, 2H)
Example 22: Preparation of (2S, 3R, 4S)-6-nitro-3,4-
dihydro-3-hydroxy-2-dimethoxymethyl-2-methyl-4-(2,3-
dihydro-2-thioxo-lH-benzimidazol-l-yl)-2H-l-benzopyran
Reaction of 300 mg (0.77 mmol) of the compound
prepared in the step 1 of the example 1 with 197 mg
(0.85 mmol) of di-2-pyridyl thiocarbonate and 9 mg
(0.08 mmol)of 4-dimethylaminopyridine was performed in
analogy to the procedure described in the example 18,
to give 240 mg (yield: 77%) of the target compound.
XH NMR (300 MHz, CDC13+1 drop DMSO) 5 1.65(s, 3H) ,
3.57(s, 3H) , 3.60(s, 3H) , 4.34(t, 1H) , 4.69(s,
6.35(d, 1H), 6.90-7.15(m, 4H), 7.27(m, 2H), 7.71(s,
8.13(dd, 1H), 11.7(br, 1H)
Mass : 431(M+), 353, 338, 206, 190
Example 23: Preparation of (3R, 4S)-6-nitro-3,4-
dihydro-3-hydroxy-2,2-dimethyl-4-(2, 3-dihydro-2-thioxo-
IH-benzimidazol-l-yl)-2H-l-benzopyran
Reaction of 400 ITK) (1.21 mmol) of the compound
prepared in the step 1 of the example 20 with 280 mg
(1.21 mmol) of di-2-pyridyl thiocarbonate was performed
in analogy to the procedure described in the example 22,
to give 250 mg (yield: 56%) of the target compound.
TH NMR (200 MHz, CDC13) 6l.48(s, 3H) , 1.60(s, 3H) ,
3.17(br-s, 1H, OH), 4.26(m, 1H), 6.28(d, 1H), 6.80(d,
1H), 6.96(m, 4H), 7.71(s, 1H) , 8.11(dd, 1H), 10.32(s,
1H, NH)
Example 24: Preparation of (3R, 4S)-6-amino-3,4-
dihydro-3-hydroxv-2,2-dimethyl-4-(2,3-dihvdro-2-thioxolH-
benzimidazol-l-yl)-2H-l-benzopyran
Reaction 145 mg (0.39 mmol) of the compound
prepared in the example 23 was performed in analogy to
the procedure described in the example 5, to give 115
mg (yield: 86%) of the target compound.
1H NMR (200 MHz, CDC13) 6l.24(s, 3H) , 1.39(s, 3H),
•3.98(m, 2H), 4.48(br-s, 1H), 5.26(d, 1H), 5.50(d, 1H),
5.71(s, 2H) , 6.02(s, 1H) , 6.32(m, 2H) , 6.77(m, 2H) ,
10.90(s, 1H)
Mass : 341, 320, 204, 106
Example 25: Preparation of (2S, 3R, 4S)-6-nitro-3,4-
dihydro-3-hydroxy-2-diethoxymethyl-2-methyl-4-(2,3-
dihydro-2-cyanoimino-lH-benzimidazol-l-yl)-2H-1-
benzopyran
Preparation of (2S, 3R, 4S)-6-nitro-3,4-
dihydro-3-hydroxy-2-diethoxymethyl-2-methyl-4-[(2-
aminophenyl)amino]-2H-l-benzopyran
Reaction of 708 mg (2.29 mmol) of epoxide compound
(2S, 3R, 4R)-6-nitro-3,4-dihydro-3,4-epoxy-2-diethoxy-
2-methyl-2H-l-benzopyran with 495 mg (4.58 mmol) of
1,2-phenylinediamine was performed in analogy to the
procedure described in the example 1, to give 791 mg
(yield: 83%) of the target compound.
1R NMR (200 MHz, CDC13) 6l.l2(t, 3H), 1.20(t, 3H),
1.51(s, 3H), 3.62(m, 2H) , 3.65(br-s, 2H, NH2), 3.81(m,
2H) , 3.92(d, 1H) , 4.08(m, 2H, NH, OH), 4.62(s, 1H) ,
4.69(dd, 1H), 6.74-6.90(m, 4H) , 6.93(d, 1H), 8.08(dd,
1H), 8.34(d, 1H)
Preparation of (2S, 3R, 4S)-6-nitro-3,4-
dihydro-3-hydroxy-2-diethoxymethyl-2-methyl-4-(2,3-
dihydro-2-cyanoimino-lH-benzimidazol-l-yl)-2H-1-
benzopyran
Reaction of 200 mg (0.48 mmol) of the compound
prepared in the above step 1 was performed in analogy
to the procedure described in the step 2 of the example
1, to give 51 mg (yield: 23%) of the target compound.
:H NMR (300 MHz, CDC13) 6l.l2(t, 3H), 1.29(t, 3H),
1.63(s, 3H), 3.59(m, 2H) , 3.67(d, 1H) , 3.83(m, 2H) ,
4.22(dd, 1H), 4.72(s, 1H) , 6.18(d, 1H) , 6.28(d,
6.92(dd, 1H) , 7.04(d, 1H), 7.13(dd, 1H), 7.36(d,
7.78(d, 1H), 8.15(dd, 1H)
Example 26: Preparation of (2S, 3R, 4S)-6-nitro-3,4-
dihydro-3-hydroxy-2-methoxymethyl-2-methyl-4-(2,3-
dihydro-2-cyanoimino-lH-benzimidazol-l-yl)-2H-1-
benzopyran
Preparation of (2S, 3R, 4S)-6-nitro-3,4-
dihydro-3-hydroxy-2-methoxymethyl-2-methyl-4-[(2-
aminophenyl)amino]-2H-l-benzopyran
Reaction of 300 mg (1.19 mmol) of epoxide compound
(2S, 3R, 4R)-6-nitro-3,4-dihydro-3,4-epoxy-2-
methoxymethyl-2-methyl-2H-l-benzopyran with 258 mg
(2.39 mmol) of 1,2-phenylinediamine was performed in
analogy to the procedure described in the step 1 of the
example 1, to give 364 mg (yield: 85%) of the target
compound.
:H NMR (300 MHz, CDC13) 6l.48(s, 3H), 3.38(s, 3H),
3.69(dd, 2H), 3.95(d, 1H) , 4.72(d, 1H) , 6.71-6.88(m,
4H), 6.95(d, 1H), 8.07(dd, 1H), 8.29(d, 1H)
Mass : 359, 296, 256, 119, 107, 80
Preparation of (2S, 3R, 4S)-6-nitro-3,4-
dihydro-3-hydroxy-2-methoxymethyl-2-methyl-4-(2, 3-
dihydro-2-cyanoimino-lH-benzimidazol-l-yl)-2H-l-
benzopyran
Reaction of 279 mg (0.78 mmol) of the compound
prepared in the above step 1 was performed in analogy
to the procedure described in the step 2 of the example
1, to give 183 mg (yield: 58%) of the target compound.
XH NMR (300 MHz, CDC13) 6l.65(s, 3H), 3.41(s, 3H),
3.76(dd, 2H), 4.26(d, 1H) , 6.10(d, 1H) , 6.30(d,
6.89(d, 1H), 7.03(d, 1H), 7.27(m, 2H) , 7.73(d,
8.14(dd, 1H), 11.88(br-s, 1H)
Mass : 409, 346, 206, 158, 132, 57
Example 27: Preparation of (2S, 3R, 4S)-6-amino-3,4-
dihydro-3-hydroxy-2-methoxymethyl-2-methyl-4-(2,3-
dihydro-2-cyanoimino-lH-benzimidazol-l-yl)-2H-1-
benzopyran
Reaction of 141 mg (0.35 mmol) of the compound
prepared in example 26 was performed in analogy to the
procedure described in the example 5, to give 38 mg
(yield: 29%) of the target compound.
1H NMR (300 MHz, CDC13) 6l.65(s, 3H) , 3.34(br-s,
1H), 3.44(s, 3H), 3.56(d, 1H) , 3.89(d, 1H) , 4.23(d, 1H) ,
5.84(d, 1H), 6.19(d, 1H) , 6.49(d, 1H) , 6.60(dd,
6.77(d, 1H), 6.93(t, 1H) , 7.09(t, 1H), 7.30(d,
Mass : 379, 319, 287, 133, 121
Example 28: Preparation of (2S, 3R, 4S)-6-nitro-3,4-
dihydro-3-hydroxy-2-methoxymethyl-2-methyl-4-(2,3-
dihydro-2-cyanoimino-lH-6-methylbenzimidazol-l-yl)-2H-
1-benzopyran
Preparation of (2S, 3R, 4S)-6-nitrc-3,4-
dihydro-3-hydroxy-2-methoxymethyl-2-methyl-4-[(2-amino-
5-methylphenyl)amino]-2H-l-benzopyran
Reaction of 300 mg (1.19 mmol) of epoxide compound
(2S, 3R, 4R)-6-nitro-3,4-dihydro-3,4-epoxy-2-
methoxymethyl-2-methyl-2H-l-benzopyran with 146 mg
(1.19 mmol) of 3,4-diaminotoluene was performed in
analogy to the procedure described in 44%) cf the
target compound.
1H NMR (300 MHz, CDC13) 5l.46(s, 3H), 2.26(s, 3H) ,
3.36(s, 3H), 3.66(m,3H), 3.37(d, 1H), 4.66(t, 1H) ,
6.52-6.73(m, 3H) , 6.95(dd, 1H), 8.04(dd, 1H) , 8.30(dd,
1H)
Mass : 373, 310, 146, 133, 121, 83
Preparation of (2S, 3R, 4S)-6-nitro-3,4-
dihydro-3-hydroxy-2-methoxymethyl-2-methyl-4-(2,3-
dihydro-2-cyanoimino-lH-6-methylbenzimidazol-l-yl)-2H-
1-benzopyran
Reaction of 144 mg (0.39 mmol) of the compound
prepared in the above step 1 was performed in analogy
to the procedure described in the step 2 of the example
1, to give 54 mg (yield: 33%) of the target compound.
1H NMR (300 MHz, CDC13) 5l.65(s, 3H), 3.27(s, 3H),
3.41(s, 3H), 3.77(dd, 2H), 4.26(dd, 1H), 6.60(dd,
6.16(d, 1H) , 7.06(m, 2H) , 7.16(d, 1H) , 7.73(d,
8.13(dd, 1H)
Mass : 423, 380, 335, 289, 172, 147, 57
Example 29: Preparation of (25, 3R, 4S)-6-nitro-3,4-
dihydro-3-hydroxy-2-([1,3]dioxan-2-yl)-2-methyl-4-(2,3-
dihydro-2-cyanoimino-lH-benzimidazol-l-yl)-2H-1-
benzopyran
Preparation of (2S, 3R, 4S)-6-nitro-3,4-
dihydro-3-hydroxy-2-([1,3]dioxan-2-yl)-2-methyl-4-[(2-
aminophenyl)amino]-2H-l-benzopyran
Reaction of 400 mg (1.36 mmol) of epoxide compound
(2S, 3R, 4R)-6-nitro-3,4-dihydro-3,4-epoxy-2-
([1,3]dioxan-2-yl)-2-methyl-2H-l-benzopyran with 295 mg
(2.73 mmol) of 1,2-phenylenediamine was performed in
analogy to the procedure described in the step I of the
example 1, to give 500 KK) (yield: 92%) of the target
compound.
*H NMR (300 MHz, CDC13) 6l.41(d, 1H) , 1.50(8, 3H) ,
2.14(m, 1H), 3.35(br-s, 2H) , 3.81(m, 2H) , 3.87(br-s,
1H), 4.11(m, 1H), 4.19(m, 2H) , 4.69(d, 1H), '4.85(3, 1H) ,
6.71-6.88(m, 4H) , 7.00(d, 1H) , 8.07(dd, 1H) , 8.32(d,
1H)
Preparation of (2S, 3R, 4S) -6-nitro-3, 4-
dihydro-3-hydroxy-2- ( [1, 3] dioxan-2-yl) -2-methyl-4- (2, 3-
dihydro-2-cyanoimino-lH-benzimidazol-l-yl) -2H-1-
benzopyran
Reaction of 400 mg (1.0 mmol) of the compound
prepared in the above step 1 was performed in analogy
to the procedure described in the step 2 of the example
1, to give 269 mg (yield: 60%) of the target compound.
ltt NMR (300 MHz, CDC13) 6l.46(d, 1H) , 1.70(s, 3H) ,
2.22(m, 1H) , 3.81(m, 2H) , 4.25(m, 3H) , 4.94(s,
6.27(d, 1H) , 6.31(d, 1H) , 6.91(dd, 1H) , 7.06(d,
7.11(dd, 1H) , 7.28(d, 1H) , 7.75(d, 1H) , 8.13(dd,
Example 30: Preparation of (2S, 3R, 4S) -6-amino-3, 4-
dihydro-3-hydroxy-2- ( [1, 3] dioxan-2-yl) -2-methyl-4- (2,3-
dihydro-2-cyanoimino-lH-benzimidazol-l-yl)-2H-1-
benzopyran
Reaction of 173 mg (0.38 mmol) of the compound
prepared in example 29 was performed in analogy to the
procedure described in the example 12, to give 25 mg
(yield: 15%) of the target compound.
1E NMR (300 MHz, CDC13) 5l.42(d, 1H), 1.71(s, 3H),
2.18(m, 1H) , 3.50(d, 2H), 3.75(dd, 1H), 3.92)dd,
4.18-4.31(m, 3H), 5.00(s, 1H), 5.90(d, 1H), 6.18(d,
6.47(d, 1H) , 6.60(dd, 1H) , 6.83(d, 1H), 6.92(dd,
7.08(dd, 1H), 7.23(d,'lH), 11.86(br-s, 1H)
Mass : 421 (M+)
Example 31: Preparation of (2S, 3R, 4S)-6-nitro-3,4-
dihydro-3-hydroxy-2-([1,3]dioxolan-2-yl)-2-methyl-4-
(2,3-dihydro-2-cyanoimino-lH-benzimidazol-l-yl)-2H-1-
benzopyran
Preparation of (2S, 3R, 4S)-6-nitro-3,4-
dihydro-3-hydroxy-2-([1,3]dioxolan-2-yl)-2-methyl-4-
[(2-aminophenyl)amino]-2H-l-benzopyran
Reaction of 417 mg (1.49 mmol) of epoxide compound
(2s, 3R, 4R)-6-nitro-3,4-dihydro-3,4-epoxy-2-
([l,3]dioxolan-2-yl)-2-methyl-2H-l-benzopyran with 323
mg (3.00 mmol) of 1,2-phenylinediamine was performed in
analogy to the procedure described in the step 1 of the
example 1, to give 596 mg (yield: 87%) of the target
compound.
XH NMR (300 MHz, CDC13) 51.54(3, 3H) , 3.40(br-s,
3H) , 3.80-4.08(m, 6H), 4.11(m, 1H), 4.86(d, 1H),
5.26(s, 1H), 6.75-6.87(m, 4H) , 6.95(d, 1H) , 8.06(dd,
1H) , 8.27(d, 1H)
Preparation of (2S, 3R, 4S)-6-nitro-3,4-
dihydro-3-hydroxy-2-([1,3]dioxolan-2-yl)-2-methyl-4-
(2, 3-dihydro-2-cyanoimino-lH-benzimidazol-l-yl)-2H-1-
benzopyran
Reaction of 200 mg (0.52 mmol) of the compound
prepared in the above step 1 was performed in analogy
to the procedure described in the step 2 of the example
1, to give 140 mg (yield: 62%) of the target compound.
*H NMR (300 MHz, CDC13) 6l.59(s, 3H), 3.53(m, 1H),
3.80(m, 1H) , 3.88(m, 1H) , 3.98(m, 1H) , 4.40(d,
5.39(s, 1H), 6.24(d, 1H) , 6.53(d, 1H) , 7.00(dd,
7.08(d, 1H) , 7.18(dd, 1H), 7.33(dd. 1H) , 7.55(d,
8.10(dd, 1H)
Mass : 437 (M+)
Example 32: Preparation of (2S, 3R, 4S) -6-nitro-3, 4-
dihydro-3-hydroxy-2- ([1,3] -5, 5-dimethyldioxan-2-yl) -2-
methyl-4- (2, 3-dihydro-2-cyanoimino-lH-benzimidazol-lyl)
-2H-l-benzopyran
Preparation of (2S, 3R, 4S) -6-nitro-3, 4-
dihydro-3-hydroxy-2- ( [1, 3] -5, 5-dimethyldioxan-2-yI) -2-
methyl-4- [ (2-aminophenyl) amino] -2H-l-benzopyran
Reaction of 400 mg (1.24 mmol) of epoxide compound
(2S, 3R, 4R) -6-nitro-3,4-dihydro-3,4-epoxy-2-( [1,3]-
5, 5-dimethyldioxan-2-yl) -2-methyl-2H-l-benzopyran with
269 mg (2.49 mmol) of 1, 2-phenylenediamine was
performed in analogy to the procedure described in the
step I of the example 1, to give 474 mg (yield: 89%) of
the target compound.
:H NMR (300 MHz, CDC13) 50.75(s, 3H) , 1.19(s, 3H) ,
1.54(s, 3H), 3.36(br-s, 2H) , 3.48(m, 2H) , 3.71(m, 2H) ,
3.73(br-s, 1H), 3.91(br-s, 1H) , 4.14(d, 1H) , 4.70(br-s,
1H), 4.75(s, 1H), 6.71-6.88(m, 4H) , 7.00{d, 1H) ,
8.08(dd, 1H), 8.33(d,
Preparation of (2S, 3R, 4S) -6-nitro-3, 4-
dihydro-3-hydroxy-2- ([1,3] -5, 5-dimethyldioxan-2-yl) -2-
methyl-4-(2,3-dihydro-2-cyanoimino-lH-benzimidazol-lyl)-
2H-l-benzopyran
Reaction of 200 mg (0.47 mmol) of the compound
prepared in the above step 1 was performed in analogy
to the procedure described in the step 2 of the example
1, to give 178 mg (yield: 79%) of the target compound.
1H NMR (300 MHz, CDC13) 50.77(s, 3H), 1.33(s, 3H) ,
1.72(s, 3H), 3.46(d, 1H), 3.52(d, 1H), 3.64(br-s,
3.76(d, 2H), 4.26(d, 1H) , 4.83(s, 1H) , 6.28(d,
6.31(d, 1H), 6.91(dd, 1H), 7.10(m, 2H) , 7.34{d,
7.76(d, 1H), 8.13(dd, 1H)
Mass : 479 (M+)
The following experiments were performed to
investigate pharmacological activities of compounds of
the present invention represented in .
Experiment I: Vasodilating effect on blood vessel
isolated from white rats
In order to investigate vasorelaxation effect of
compounds represented in on blood vessel,
following experiments were performed.
White rats (350 ~ 450 g; the Experimental Animal
Team of the Korea Research Institute of Chemical
Technology) were knocked to be unconscious by hitting
the occipital region, sacrificed by cervical
dislocation, and underwent thoracotomy. After being
quickly removed, the thoracic aorta was deprived of the
adipose tissue and cut into aortic rings of 3 nun width.
The aorta was lightly rubbed with cotton club soaked in
a modified Krebs Henseleit buffer (physiological salt
solution) to remove the inner epithelial layer
therefrom. While being hung in an organ bath
containing a physiological buffer, the vascular smooth
muscle was allowed to equilibrate under a resting
tension of 2 g and then, stand 'for 1 hour at 37°C for
stabilization, supplying a carbogen consisting of 95%
02 I 5% COo. Thereafter, the vascular smooth muscle was
constricted with 1CT5 M phenylephrine and washed several
times with physiological saline solution. The said
procedure was repeated to ensure the stable reactivity
of vascular smooth muscle to repetitive
constriction/relaxation.
Thereafter, 3 x 1CT6 M methoxamine was applied to
induce an intensive constriction in the vascular smooth
muscle. When the vasoconstriction induced by the
methoxamine was reached and maintained to a maximum,
test compounds and control material were cumulatively
added to the organ bath with concentration of 1, 3, 10
and 30 M M, respectively, to examine the vasodilating
effect. Cromakalim and BMS-180448 (compound of
) , known to be the first generation KATP
opener with potent vasodilating effect, were used as
control materials.
The change rate of constriction by the addition
of the drugs compared to the maximal constriction
induced by methoxamine was calculated to plot a
concentration-relaxation response curve. Through a
least linear regression analysis, IC50 that the drug
concentration at which the vascular tissue is relaxed
to 50% extent of the maximal constriction, was obtained
for each drug. And the results were shown in the below
Table 1.

Vasodilating effect of the compounds of
(Table Removed)

As shown in rhe above Table I, Cromakalim
represented a potent vasorelaxation effect, showing
0.067 nM of IC50 on the isolated rat aorta constricted
with methoxamine (3 nM) while IC50 of BMS-180448 was
1.38 nM, which was twenty times weaker vasorelaxation
effects than that of Cromakalim. On the other hand,
IC50 of the compound of example 1 was 50.1 jiM, showing
36 times weaker vasorelaxation effect than that of BMS-
180448, a general KATP opener, used as a control
material. The compound of example 20 also showed 10
times weaker vasorelaxation effect than that of BMS-
180448.
Cromakalim or BMS-180448 protects heart by acting
toward KATP in heart and drops blood pressure by
dilating blood vessels by acting toward KATP in coronary
and peripheral blood vessels. Hypotensive action may
mask any cardioprotective effects due to reduction
under coronary artery perfusion pressure, and would
limit utility in treating myocardial ischemia.
Therefore, the compounds of the present invention may
be more optimal for cardioprotective agents by virtue
of their weak vasorelaxation activity.
Again, the compounds of the present invention
have enhanced cardioprotectjve function with weak
Experiment 2: Cardioprotective effect on isolated
ischemic heart models of white rats
The experiment confirming that the compounds of
have the protective effect (antiischemic
effect) on ischemic heart was accomplished in the below.
100 mg/kg of sodium pentobarbital was injected in
abdominal cavity of white male rats (300 ~ 450 g; the
experimental animal team of the Korea Research
Institute of Chemical Technology) to anesthetize them.
Then, an intravenous injection of 1000 U/kg of heparin
was performed before taking out heart. Particularly,
cannula(PE 240) was inserted in the trachea, and
artificial respiration was tried upon the rat by using
a rodent ventilator. Under that condition, aortic
cannula was inserted in the aorta and heart was taken
out under retrograde perfusion. The extracted heart
was hung on Langendorff apparatus quickly and
unnecessary tissues on heart were removed. Perfusion
was induced under static pressure (85 mmHg) with 37°C
modified Krebs-Henseleit bicarbonate buffer
(composition : 116 NaCl, 4.7 KC1, 1.1 MgS04, 1.17
KH2P04, 24.9 NaHC03, 2.52 CaCl2, 8.32 Glucose, 2.0
Pyruvate) saturated with 95% O2/5% C02. A metal cannula,
to which a latex balloon filled with a ethanoldistilled
water mixture (1:1 vol/vol) was linked, was
inserted in left ventricle through pulmonary vein.
Then, left ventricular pressure transmitted through the
balloon was transduced by using pressure transducer,
and amplified by using isovolumetric amplifier(Plugsys
bridge amplifier). Then, the pressure was recorded in
a recorder (Linearcorder mark 8 WR 3500). Thereafter,
heart was stabilized for 15 minutes. Then, left
ventricular end diastolic pressure (LVEDP) was given by
5 mmHg and such volume of the balloon was kept
throughout the experiments.
Baseline cardiac contractile function, heart rate
(HR) , and coronary flow (CF) were measured. Cardiac
contractile function was calculated by subtracting LVSP
(left ventricular peak systolic pressure) from LVEDP
(left ventricular end diastolic pressure), yielding
LVDP (left ventricular developed pressure). Double
product RPP (rate-pressure product)(DP), another
important parameter for indirectly assessing cardiac
performance in Langendorff heart, in which cardiac
output could not be measured ordinarily, was calculated
by multiplying HR by LVDP. Throughout the experiment,
total coronary blood flow was measured by the use of
coronary flow probe (diameter: 1.0 mm) installed in
aortic cannula with electromagnetic flowmeter.
Temperature of heart was steadily maintained by
immersing the heart at 37°C in physiological saline
solution to which 95% 02/5% CO2 was constantly supplied.
After stabilization for 15 min, the hearts were pretreated
for 10 min with vehicle (0.04% DMSO) only or a
compound of the present invention or the control
material in the vehicle. Thereafter, cardiac
contractile function, HR and CF were repeatedly
measured. Global ischemia was induced by completely
shutting off the perfusate for 30 min. Severity of
ischemia was determined as the time to contracture (TTC,
min) during global ischemia in which the first 5 mmHg
increase in EDP was observed. Then, the hearts were
reperfused and, 30 min later, contractile functions
(LVDP, HR and CF) were repeatedly measured. After
reperfusion was accomplished for 30 min, LDH (lactate
dehydrogenase) was measured with a kit as a sensitive
index for loss of cell viability. The results were
shown in Table 2.

Cardioprotective effect in ischemic heart models of
Rats
(Table Removed)

In vehicle-treated group, reperfusion DP (LVDP X
HR) , an index for contractility function, was decreased
to 15.8% of pre-treatment DP, and EDP was increased to
45.1 mmHg from 5 mmHg, and TTC was 19.8 min, and
reperfusion LDH release was 31.3 unit/g as shown in the
above.
In BMS-180448 treated group, reperfusion
contractile function (DP, LVDP x HR) was 67.6% of pretreatment
DP, which was significantly improved compared
to the vehicle treated group. EDP was 16.5 mmHg,
significantly lower than control, and TTC was 27.8 min,
prolonged than control, and reperfusion LDH release was
17.2 Unit/g, decreased than control. Then, in BMS-
180448 treated group, all parameters showed significant
protective effect on ischemic heart.
When compared only in antiischemic effects based
on those parameters, cardiac contractile function, EDP,
TTC, and LDH release, the compounds of the present
invention were similar to or superior to BMS-180448.
However, because the compounds of the present invention
have remarkably lower vasorelaxant effects than BMS-
180448 dose, they are far superior to the conventional
drugs in cardioselective antiischemic activity.
Especially, the compound of Example 1 showed a .good
cardioprotective effect, of which contractile function
(LVDP x HR) was improved to 68.8% of pre-treatment
index, and EDP was 20.3 mmHg, and TTC was 23.2 min, and
reperfusion LDH release was 8.1 unit/g, with very low
vasodilation activity (IC50 = 50.1 p M) . So, it shows
much better cardioselectivity upon vasodilation than
BMS-180448. Consequently, the compounds of the present
invention can be used for the treatment of ischemic
heart diseases by virtue of their excellent selectivity
and protective activity against ischemic cardiovascular
diseases. Besides, the compounds can also be used as a
protective agent for ischemic brain and retinal cell
damage caused by ischemia-reperfusion or for storage
organs,
Experimental 3: Acute oral toxicity test in rats
The following experiments were performed to see
if the compounds of had acute toxicity in
rats .
6-week old SPF SD line rats were used in the
tests for acute toxicity test. The compounds prepared
in the Example 1 were suspended in 0.5% methylcelLulose
solution and orally administered once to 2 rats per
group with the dosage of 1 g/kg/15 M. Death, clinical
symptoms, and weight change in rats were observed,
hematological tests and biochemical tests of blood were
performed, and any abnormal signs in the
gastrointestinal organs of chest and abdomen were
checked with eyes during autopsy.
The results showed that the test compounds did
not cause any specific clinical symptoms, weight change,
or death in rats. No change was observed in
hematological tests, biochemical tests of blood, and
autopsy. The compounds used in this experimert were
evaluated to be safe substances since they did not
cause any toxic change in rats up to the level of 2
g/kg and their estimated LD50 values were much jreater
INDUSTRIAL APPLICABILITY
As explained hereinbefore, the compounds of
present invention represented in have
excellent cardioprotective effect against damage by
ischemia-reperfusion without lowering blood pressure
owing to their weak vasorelaxation activity. Thus, a
pharmaceutical composition containing benzopyran
derivatives substituted with a benzimidazole derivative,
represented in , or pharmaceutically
acceptable salts of the same can be used as a
protective or therapeutic agent for ischemiareperfusion
related damage or diseases, that is, the
compounds are not only useful for the treatment of
ischemic heart diseases such as myocardial infarction,
unstable angina pec'toris, etc, the protection of heart
from the damage caused by thrombolytics or reperfusion
therapy such as PTCA (percutaneous transluminal
coronary angioplasty) and CABG (coronary artery bypass
graft), and the protection of ischemia-reperfusion
related tissues such as nerve cells, brain, retinal
cells, storage organs, etc.
Those skilled in the art will appreciate that the
conceptions and specific embodiments disclosed in the
foregoing description may be readily utilized as a
basis for modifying or designing other embodiments for
carrying out the same purposes of the present invention.
Those skilled in the art will also appreciate that such
equivalent embodiments do not depart from the spirit
and scope of the invention as set forth in the appended claims.


We claim:
1. A bcnzopyran derivatives substituted with a benzimidazole derivative of Formula 1:

(Formula Removed)

Wherein,
X is O,S orNCN;
R1 is N02, NH2, H, CN, NHCOCH3, NHCOCF3 or NHSO2CH3;


and
(Formula Removed)
wherein,
Ra is C1 □C4 straight or branched alkyl; and
Z is C2 □C6 straight or branched alkyl;
R3 is OH or OCOCH3;
R4 is C1 □C4 straight or branched alkyl;
R5 and R6 are independently H, C1 □ C4 straight or branched alkyl, alkoxy or halogen;
* represents a chiral carbon,
or a pharmaceutically acceptable salt thereof.


2. The compound as claimed in claim 1, selected from the group consisting of:
1 )(2S, 3R, 4S)-6-nitro-3,4-dihydro-3-hydroxy-2-dimethoxymethyl-2-methyl-4-(2,3-dihydro-
2-cyanoimino-1 H-benzimidazol-1 -yl)-2H-1 -benzopyran; 2)(2R, 3R, 4S)-6-nitro-3,4-dihydro-3-hydroxy-2-dimethoxymethyl-2-methyl-4-(2,3-dihydro-
2-cyanoimino-1 H-benzimidazol-1 -yl)-2H-1 -benzopyran; 3)(2S, 3S, 4R)-6-nitro-3,4-dihydro-3-hydroxy-2-dimethoxymethyl-2-methyl-4-(2,3 -dihydro-
2-cyanoimino-1 H-benzimidazol-1 -yl)-2H-1 -benzopyran; 4)(2R, 3S,4S)-6-nitro-3,4-dihydro-3-hydroxy-2-dimethoxymethyl-2-methyl-4-(2,3-dihydro-
2-cyanoimino-1 H-benzimidazol-1 -yl)-2H-1 -benzopyran; 5)(2S, 3R,4S)-6-amino-3,4-dihydro-3-hydroxy-2-dimethoxymethyl-2-methyl-4-(2,3-
dihydro-2-cyanoimino-1 H-benzimidazol-1 -yl)-2H-1 -benzopyran; 6)(2S, 3R, 4S)-6-nitro-3,4-dihydro-3-hydroxy-2-dimethoxymethyl-2-methyl-4-(2,3-dihydro-
3-methyl-2-cyanoimino-1 H-benzimidazol-1 -yl)-2H-1 -benzopyran; 7)(2S, 3R,4S)-6-nitro-3,4-dihydro-3-hydroxy-2-dimethoxymethyl-2-methyl-4-(2,3-dihydro-
5,6-dimethyl-2-cyanoimino-1 H-benzimidazol-1 -yl)-2H-1 -benzopyran; 8)(2S, 3R,4S)-6-amino-3,4-dihydro-3-hydroxy-2-dimethoxymethyl-2-methyl-4-(2,3-
dihydro-5,6-dimethyl-2-cyanoimino-1 H-benzimidazol-1 -yl)-2H-1 -benzopyran; 9)(2S, 3R, 4S)-6-nitro-3,4-dihydro-3-hydroxy-2-dimethoxymethyl-2-methyl-4-(2,3-dihydro-
3,5,6-trimethyl-2-cyanoimino-1 H-benzimidazol-1 -yl)-2H-1 -benzopyran; 10)(2S, 3R, 4S)-6-amino-3,4-dihydro-3-hydroxy-2-dimethoxymethyl-2-methyl-4-(2,3-
dihydro-3,5,6-trimethyl-2-cyanoimino-1 H-benzimidazol-1 -yl)-2H-1 -benzopyran; 11)(2S, 3R, 4S)-6-nitro-3,4-dihydro-3-acetoxy-2-dimethoxymethyl-2-methyl-4-(2,3-dihydro-
2-cyanoimino-1 H-benzimidazol-1 -yl)-2H-1 -benzopyran; 12)(2S, 3R, 4S)-6-amino-3,4-dihydro-3-acetoxy-2-dimethoxymethyl-2-methyl-4-(2,3-
dihydro-2-cyanoimino-1 H-benzimidazol-1 -yl)-2H-1 -benzopyran; 13)(2S, 3R, 4S)-6-acetamino-3,4-dihydro-3-hydroxy-2-dimethoxymethyl-2-methyl-4-(2,3-
dihydro-2-cyanoimino-1 H-benzimidazol-1 -yl)-2H-1 -benzopyran;
14)(2S, 3R, 4S)-6-acetamino-3,4-dihydro-3-acetoxy-2-dimethoxymethyl-2-methyl-4-(2,3-dihydro-2-cyanoimino-1 H-benzimidazol-1 -yl)-2H-1 -benzopyran;
15)(2S, 3R, 4S)-6-benzoylamino-3,4-dihydro-3-hydroxy-2-dimethoxymethyl-2-methyl-4-(2,3-dihydro-2-cyanoimino-1 H-benzimidazol-1 -yl)-2H-1 -benzopyran;
16)(2S, 3R, 4S)-6-(trifluoroacetyl)amino-3,4-dihydro-3-hydroxy-2-dimethoxymethyl-2-methyl-4-(2,3 -dihydro-2-cyanoimino-1 H-benzimidazol-1 -yl)-2H-1 -benzopyran;

17)(2S, 3R, 4S)-6-methanesulfonylamino-3,4-dihydro-3-hydroxy-2-dimemoxymethyl-2-
methyl-4-(2,3 -dihydro-2-cyanoimino-1 H-benzimidazol-1 -yl)-2H-1 -benzopyran; 18)(2S, 3R,4S)-6-nitro-3,4-dihydro-3-hydroxy-2-dimethoxymethyl-2-methyl-4-(2,3-dihydro-
2-oxo-1 H-benzimidazol-1 -yl)-2H-1 -benzopyran; 19)(2S, 3R, 4S)-6-amino-3,4-dihydro-3-hydroxy-2-dimethoxymethyl-2-methyl-4-(2,3-
dihydro-2-oxo-1 H-benzimidazol-1 -yl)-2H-1 -benzopyran; 20)(2S, 3R, 4S)-6-nitro-3,4-dihydro-3-hydroxy-2-dimethoxymethyl-2-methyl-4-(2,3-dihydro-
2-thioxo-1 H-benzimidazol-1 -yl)-2H-1 -benzopyran; 21)(2S, 3R,4S)-6-nitro-3,4-dihydro-3-hydroxy-2-diethoxymethyl-2-methyl-4-(2,3-dihydro-2-
cyanoimino-1 H-benzimidazol-1 -yl)-2H-1 -benzopyran; 22)(2S, 3R,4S)-6-nitro-3,4-dihydro-3-hydroxy-2-methoxymethyl-2-methyl-4-(2,3-dihydro-2-
cyanoimino-1 H-benzimidazol-1 -yl)-2H-1 -benzopyran; 23)(2S, 3R,4S)-6-amino-3,4-dihydro-3-hydroxy-2-methoxymethyl-2-methyl-4-(2,3-dihydro-
2-cyanoimino-1 H-benzimidazol-1 -yl)-2H-1 -benzopyran; 24)(2S, 3R, 4S)-6-nitro-3,4-dihydro-3-hydroxy-2-methoxymethyl-2-methyl-4-(2,3-dihydro-2-
cyanoimino-1 H-6-methylbenzimidazol-1 -yl)-2H-1 -benzopyran; 25)(2S, 3R, 4S)-6-nitro-3,4-dihydro-3-hydroxy-2-([l ,3]dioxan-2-yl)-2-methyl-4-(2,3-
dihydro-2-cyanoimino-1 H-benzimidazol-1 -yl)-2H-1 -benzopyran; 26)(2S, 3R, 4S)-6-amino-3,4-dihydro-3-hydroxy-2-([l ,3]dioxan-2-yl)-2-methyl-4-(2,3-
dihydro-2-cyanoimino-1 H-benzimidazol-1 -yl)-2H-1 -benzopyran; 27)(2S, 3R, 4S)-6-nitro-3,4-dihydro-3-hydroxy-2-([l,3]dioxolan-2-yl)-2-methyl-4-(2,3-
dihydro-2-cyanoimino-1 H-benzimidazol-1 -yl)-2H-1 -benzopyran; and 28)(2S, 3R, 4S)-6-nitro-3,4-dihydro-3-hydroxy-2-([l,3]-5,5-dimethyldioxan-2-yl)-2-methyl-
4-(2,3-dihydro-2-cyanoimino-1 H-benzimidazol-1 -yl)-2H-1 -benzopyran,
or a pharmaceutically acceptable salt thereof.
3. A compound as claimed in claim 1 as and when used in the preparation of a pharmaceutical composition for treatment of ischemia-reperfusion related brain injury, retinal cells or storage organs, and protection of storage organs such as heart, kidney, liver and tissues along with carriers.

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5405-DELNP-2005-Abstract-(04-03-2009).pdf

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5405-DELNP-2005-Claims-(04-03-2009).pdf

5405-DELNP-2005-Claims-(21-04-2009).pdf

5405-delnp-2005-claims.pdf

5405-DELNP-2005-Correspondence-Others-(04-03-2009).pdf

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5405-delnp-2005-correspondence-others.pdf

5405-delnp-2005-description (complete).pdf

5405-DELNP-2005-Form-1-(04-03-2009).pdf

5405-delnp-2005-form-1.pdf

5405-delnp-2005-form-18.pdf

5405-DELNP-2005-Form-2-(04-03-2009).pdf

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5405-DELNP-2005-Form-3-(04-03-2009).pdf

5405-delnp-2005-form-3.pdf

5405-delnp-2005-form-5.pdf

5405-delnp-2005-pct-304.pdf

5405-DELNP-2005-Petition-138-(21-04-2009).pdf


Patent Number 234548
Indian Patent Application Number 5405/DELNP/2005
PG Journal Number 26/2009
Publication Date 26-Jun-2009
Grant Date 05-Jun-2009
Date of Filing 24-Nov-2005
Name of Patentee DONGBU HANNONG CHEMICAL CO., LTD.
Applicant Address 19-20F, DONGBU FINANCIAL CENTER, 891-10 DAECHI-DONG, KANGNAM-KU, SEOUL 135-523 KOREA.
Inventors:
# Inventor's Name Inventor's Address
1 LIM HONG #104-203 BANPO APT., 952 BANPO-DONG, SEOCHO-KU, SEOUL 137-041 KOREA.
2 LEE DONG HA #110-401 SEJONG APT., JEONMIN-DONG, YUSCONG-KU 305-728, TACJON-SI 305-728 KOREA.
3 KIM SUN OK #102-706 DURE APT., SHINSUNG-DONG, YUSCONG-KI, TAEJON-SI 305-345 KOREA.
4 YOO SUNG-EUN 314-6 KEUMAM-LI, JANGGI-MYUN, KONGJU-SI, CHUNGCHONGNAM-DO 314-911 KOREA.
5 YI KYU YANG #130-1001 HANBIT APT., OUN-DONG, YUSEONG-KU, TAEJON-SI, 305-755 KOREA.
6 KIM NAK JEONG #102-701 HANWOOL APT., SHINSUNG-DONG, YUSEONG-KU, TAEJON-SI 305-345 KOREA.
7 KWANG SUN KYUNG 107-1002 HANARO APT., WOLPYEONG-DONG, SEO-KU, TAEJON-SI 302-748 KOREA.
8 KIM TAE MI #205-803 JUNGONG APT., WOLPYEONG-2DONG, SEO-KU, TAEJON-SI 302-282 KOREA
9 LEE BYUNG HO #116-804 HANBIT APT., OUN-DONG, YUSEOUNG-KU, TAEJON-SI 305-755 KOREA.
10 SEO HO-WON #104-1402 TAEPYEONG APT., TAEPYEONG-DONG, JUNG-KU, TAEJON-SI 301-211 KOREA.
11 LEE SUN KYUNG #8-306 LG APT., DORYONG-DONG, YUSEONG-KU, TAEJON-SI 305-340 KOREA.
12 SUH JEE HEE #116-802 HANBIT APT., OUN-DONG, YUSEONG-KU, TAEJON-SI 305-755 KOREA.
PCT International Classification Number C07D 405/04
PCT International Application Number PCT/KR2004/001269
PCT International Filing date 2004-05-28
PCT Conventions:
# PCT Application Number Date of Convention Priority Country
1 10-2003-0034109 2003-05-28 Republic of Korea