U.S. patent application number 12/468929 was filed with the patent office on 2009-09-10 for new 2-(alpha-hydroxypentyl) benzoates, their preparations and their uses.
This patent application is currently assigned to THE INSTITUTE OF MATERIA MEDICA OF CHINESE ACADEMY OF MEDICAL SCIENCES.. Invention is credited to Ying PENG, Xiaoliang WANG, Zhibin XU, Jinghua YANG.
Application Number | 20090227809 12/468929 |
Document ID | / |
Family ID | 33569588 |
Filed Date | 2009-09-10 |
United States Patent
Application |
20090227809 |
Kind Code |
A1 |
YANG; Jinghua ; et
al. |
September 10, 2009 |
NEW 2-(ALPHA-HYDROXYPENTYL) BENZOATES, THEIR PREPARATIONS AND THEIR
USES
Abstract
The invention relates to new synthetic 2-(.alpha.-hydroxypentyl)
benzoates, their preparation and pharmaceutical compositions
containing such salts as active ingredients. The invention also
relates to the use of the compounds for preventing and treating
cardioischemia, cerebroischemia and cardiac or cerebral arterial
occlusion (obstruction), etc.
Inventors: |
YANG; Jinghua; (Beijing,
CN) ; WANG; Xiaoliang; (Beijing, CN) ; XU;
Zhibin; (Beijing, CN) ; PENG; Ying; (Beijing,
CN) |
Correspondence
Address: |
LADAS & PARRY LLP
26 WEST 61ST STREET
NEW YORK
NY
10023
US
|
Assignee: |
THE INSTITUTE OF MATERIA MEDICA OF
CHINESE ACADEMY OF MEDICAL SCIENCES.
|
Family ID: |
33569588 |
Appl. No.: |
12/468929 |
Filed: |
May 20, 2009 |
Related U.S. Patent Documents
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Application
Number |
Filing Date |
Patent Number |
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10512792 |
May 27, 2005 |
7550507 |
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PCT/CN02/00320 |
May 9, 2002 |
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12468929 |
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Current U.S.
Class: |
560/60 |
Current CPC
Class: |
A61P 43/00 20180101;
A61P 7/02 20180101; C07C 65/01 20130101; A61P 9/10 20180101; C07C
65/03 20130101 |
Class at
Publication: |
560/60 |
International
Class: |
C07C 69/78 20060101
C07C069/78 |
Claims
1. A compound of the following general formula (I): ##STR00004##
wherein M is a monovalent metal ion, a divalent metal ion, or an
organic base group and n is 1 or 2.
2. The compound according to claim 1, wherein M is a potassium ion,
a sodium ion, or a lithium ion.
3. The compound according to claim 1, wherein M is a calcium ion, a
magnesium ion, or a zinc ion.
4. The compound according to claim 1, wherein M is an anilino
group, a benzyl amino group, a morpholinyl group or a diethylamino
group.
5. A preparation method of the compound of general formula (I)
according to claim 1 wherein M is a monovalent metal ion,
comprising: solving an equivalent racemic
3-n-butyl-isobenzofuran-1-(3H)-one in a hydrolysis and ring-opening
reaction solvent medium, adding an equivalent or slightly excess
amount of monovalent base, conducting hydrolysis and ring-opening
reaction under the temperature of 10.about.100.degree. C., to
afford the 2-(.alpha.-hydroxypentyl) benzoate of general formula
(I) wherein M is a monovalent metal ion.
6. The preparation method according to claim 5, wherein the solvent
for the hydrolysis and ring-opening reaction is any one of
methanol, ethanol, acetone, isopropanol, water or mixture of
water-alcohol (ketone), the monovalent base is an inorganic base,
such as sodium hydroxide, potassium hydroxide, or lithium
hydroxide; or a chemically pure organic base, such as sodium
(potassium) methoxide, sodium (potassium) ethoxide.
7. A preparation method of the compound of general formula (I)
according to claim 1 wherein M is a divalent metal ion, comprising:
solving 2-(.alpha.-Hydroxypentyl) benzoate of general formula (I)
wherein M is a monovalent metal ion (sodium or potassium) in a
solvent, adding an equivalent or slightly excess amount of
chemically pure divalent metal salt and conducting an ion-exchange
reaction under the temperature of 10-100.degree. C., to afford the
2-(.alpha.-hydroxylpentyl) benzoate of general formula (I) wherein
M is a divalent metal ion.
8. The preparation method according to claim 7, wherein the
reaction solvent medium is any one of methanol, ethanol, acetone,
isopropanol, water or a mixture of water-alcohol (ketone); and the
divalent metal salt is any one of magnesium chloride, calcium
chloride, or zinc chloride.
9. A preparation method of the compound of general formula (I)
according to claim 1, comprising: (1) solving
2-(.alpha.-hydroxypentyl) benzoate of general formula (I) wherein M
is a monovalent metal ion in a solvent medium, adding an inorganic
acid to the solution and adjusting the pH value to 6.0.about.2.0,
allowing the 2-(.alpha.-hydroxypentyl) benzoate to react with the
acid to afford 2-(.alpha.-hydroxypentyl) benzoic acid, at a
temperature of -20.about.20.degree. C., preferably
-20.about.0.degree. C.; (2) adding an organic extraction solvent to
the so-obtained benzoic acid solution and extracting free
2-(.alpha.-hydroxypentyl) benzoic acid to obtain an organic
solution containing 2-(.alpha.-hydroxypentyl) benzoic acid, keeping
the solution for use under the temperature of -20.about.10.degree.
C., preferably -20.about.0.degree. C.; (3) adding a solution which
contains an equivalent or slightly excess amount of monovalent base
to the 2-(.alpha.-hydroxypentyl) benzoic acid solution obtained in
the above step under the temperature of -10.about.0.degree. C., to
afford a 2-(.alpha.-hydroxylpentyl) benzoate of general formula (I)
wherein M is a monovalent metal ion; (4) adding a solution which
contains an equivalent or slightly excess amount of divalent base
or a divalent metal salt solution to the 2-(.alpha.-hydroxypentyl)
benzoic acid solution obtained in step (2) under the temperature of
-10.about.0.degree. C., to afford a 2-(.alpha.-hydroxypentyl)
benzoate of general formula (I) wherein M is a divalent metal ion;
(5) adding a solution which contains an equivalent or slightly
excess amount of organic base to the 2-(.alpha.-hydroxypentyl)
benzoic acid solution obtained in step (2) under the temperature of
-10.about.0.degree. C., to afford a 2-(.alpha.-hydroxypentyl)
benzoate of general formula (I) wherein M is an organic base.
10. The preparation method according to claim 9, wherein the acid
used in the acidification reaction is any one of concentrated or
diluted hydrochloric acid or sulfuric acid; the acidification
reaction temperature is controlled within the range of
-20.about.+20.degree. C.; the organic solvent to extract
2-(.alpha.-hydroxypentyl) benzoic acid is any one of ether, ethyl
acetate, chloroform, dichloromethane, benzene, toluene, petroleum
ether, n-hexane, or cyclohexane; the monovalent base is any one of
sodium hydroxide, potassium hydroxide, lithium hydroxide, sodium
(or potassium) methoxide, or sodium (or potassium) ethoxide; the
divalent inorganic metal salt or divalent inorganic base is any one
of magnesium chloride, magnesium carbonate, calcium chloride,
calcium carbonate, zinc chloride, zinc carbonate, magnesium
sulfate, magnesium hydroxide, or calcium hydroxide; the organic
base is any one of aniline, benzyl amine, morpholine or
diethylamine; the solvent is any one of methanol, ethanol, acetone,
isopropanol, water or water-alcohol (ketone) or any one of ether,
acetyl acetate, chloroform, dichloromethane, benzene, toluene,
petroleum ether, n-hexane, or cyclohexane.
Description
TECHNICAL FIELD
[0001] This invention relates to new chemically synthetic
2-(.alpha.-hydroxypentyl) benzoates, their preparations and
pharmaceutical compositions containing the salts as active
ingredients. This invention also relates to the uses of the
compounds in the prevention and treatment of the diseases such as
cardioischemia, cerebroischemia, heart and brain arterial
occlusions, etc.
BACKGROUND OF THE INVENTION
[0002] Acute ischemic cerebral apoplexy is a common disease with
high incidence (12-18 cases per ten thousands) and high death rate
(6-12 cases per ten thousands) which seriously endangers human's
health, which often leaves sequela in the survivals and imposes a
heavy burden on the patients' family and society. Therefore, it is
very valuable of developing drug for the prevention and treatment
of the disease. Many researchers have been studying the mechanism
of the disease (acute ischemic cerebral apoplexy) since the 1980's,
and have proposed theories such as energy metabolism, excitatory
poison, oxidative injury, calcium overload and many other theories
for the purpose of developing high effective and low toxic drugs.
However, an ideal therapeutic drug is still under developing. Drugs
such as calcium antagonists, excitatory receptor antagonists, free
radical scavengers are being clinically used, but the effects are
uncertain. Thrombolytic drugs such as t-PA are being used to treat
acute ischemic cerebral apoplexy (within 6 hrs from incidence), and
is effective, but the hazard of hemorrhage has not been solved.
Thus it is still a focus of developing new drugs to treat ischemic
cerebral apoplexy.
[0003] Coronary heart disease is also a severe disease which harms
human's health. Due to the coronary atherosclerosis and the
formation of thrombus, ischemic cardiac muscle trauma is induced.
For this reason, it has been being a leading work of developing new
drugs to prevent and treat coronary atherosclerosis, prevent
thrombus formation and dilatate coronary artery.
SUMMARY OF THE INVENTION
[0004] The object of the present invention is to provide new
2-(.alpha.-hydroxypentyl) benzoates which can significantly inhibit
platelet aggregation and ameliorate cerebral microcirculation and
effective to cardioischemia, cerebroischemia, heart and brain
arterial occlusions.
[0005] Another object of the present invention is to provide a
synthetic method of 2-(.alpha.-hydroxypentyl) benzoates.
[0006] Yet another object of the present invention is to provide a
pharmaceutical composition which can prevent and treat
cardioischemia, cerebroischemia, heart and brain arterial
occlusions.
[0007] The fourth object of the present invention is to provide use
of the above compounds and pharmaceutical compositions in the
prevention and treatment of cardioischemia, cerebroischemia, heart
and brain arterial occlusions, and amelioration of cerebral
microcirculation.
[0008] The present invention provides a compound of the following
general formula (I):
##STR00001##
wherein n=1,2; M is a monovalent metal ion, such as K.sup.+,
Na.sup.+, Li.sup.+; a divalent metal ion, such as Ca.sup.2+,
Mg.sup.2+, Zn.sup.2+; or an organic basic group, such as anilino,
benzyl amino, morpholinyl or diethylamino.
[0009] The preparation method of present invention is as
following:
[0010] 1. Preparation of 2-(.alpha.-hydroxypentyl) benzoates of
general formula (I) wherein M is a monovalent metal ion:
[0011] Solving an equivalent racemic
3-n-butyl-isobenzofuran-1-(3H)-one in a hydrolysis and ring-opening
reaction solvent medium, adding an equivalent or slightly excess
amount of monovalent base. After that hydrolysis and ring-opening
reaction is conducted under the temperature of 10-100.degree. C.,
for 0.5-6 hours, to afford the 2-(.alpha.-hydroxypentyl) benzoates
of general formula (I) wherein M is a monovalent metal ion.
[0012] The solvent for the hydrolysis and ring-opening reaction may
be any one of methanol, ethanol, acetone, isopropanol, water or
mixture of H.sub.2O-alcohol (or ketone).
[0013] The crystallization solvent may be any one of methanol,
ethanol, propanol, isopropanol, acetone, acetyl acetate,
chloroform, ether, dichloromethane, benzene, toluene, petroleum
ether, or a mixture of two or three above solvents with different
proportions; monovalent base may be an inorganic base, such as a
chemically pure inorganic base, for example sodium hydroxide,
potassium hydroxide, or lithium hydroxide, etc; or a chemically
pure organic base, such as sodium (or potassium) methoxide, sodium
(or potassium) ethoxide, etc.
[0014] 2. Preparation of 2-(.alpha.-hydroxypentyl) benzoates of
general formula (I) wherein M is a divalent metal ion:
[0015] 2-(.alpha.-Hydroxypentyl) benzoate of general formula (I)
wherein M is a monovalent metal ion (e.g., sodium or potassium) is
solved in a solvent, then an equivalent or slightly excess amount
of chemically pure divalent metal salt is added and an ion-exchange
reaction is conducted under the temperature of 10-100.degree. C.
for 0.5-10 hours to afford the 2-(.alpha.-hydroxypentyl) benzoates
of general formula (I) wherein M is a divalent metal ion.
[0016] The reaction solvent medium is methanol, ethanol, acetone,
isopropanol, water or a mixture of water-alcohol (or ketone);
divalent metal salt may be magnesium chloride, calcium chloride, or
zinc chloride; the crystallization solvent may be methanol,
ethanol, isopropanol, acetyl acetate, chloroform, ether,
dichloromethane, or a mixture of two or three above solvents with
different proportions.
[0017] 3. Preparation of the 2-(.alpha.-hydroxypentyl) benzoates of
general formula (I) wherein M is an organic base:
[0018] (1) 2-(.alpha.-hydroxypentyl) benzoate of general formula
(I) wherein M is a monovalent metal ion (e.g., sodium or potassium)
is solved in a solvent, then a inorganic acid is added to the
solution to adjust pH value to 6.0-2.0 at the temperature of
-20-20.degree. C., preferably -20-0.degree. C., to acidify the
2-(.alpha.-hydroxy pentyl) benzoate and 2-(.alpha.-hydroxypentyl)
benzoic acid is obtained.
[0019] (2) After the reaction is completed, an organic extraction
solvent is added to the solution and the free acid
2-(.alpha.-hydroxypentyl) benzoic acid is extracted with common
extraction method at a temperature of -20-0.degree. C. An organic
solution containing 2-(.alpha.-hydroxypentyl) benzoic acid is
obtained and the solution is kept for use under the temperature of
-20-10.degree. C., preferably -20.about.0.degree. C.
[0020] (3) To the solution obtained in the above step, a solution
which contains an equivalent or slightly excess amount of
monovalent base to the 2-(.alpha.-hydroxypentyl) benzoic acid, for
example an alcohol solution of potassium hydroxide, is added, at
the temperature of -10.about.0.degree. C., to afford the
2-(.alpha.-hydroxylpentyl) benzoate of present invention wherein M
is a monovalent metal ion, such as potassium. After the reaction is
sufficiently conducted, with a same purification method with that
of preparation method 1, the compound of present invention, i.e.,
2-(.alpha.-hydroxylpentyl) benzoate wherein M is a monovalent metal
ion is prepared.
[0021] (4) To the solution obtained in step (2), a solution which
contains an equivalent or slightly excess amount of divalent base
or a divalent metal salt to the 2-(.alpha.-hydroxypentyl) benzoic
acid, for instance an alcohol solution of calcium hydroxide, is
added, under the temperature of -10.about.0.degree. C., to afford
the 2-(.alpha.-hydroxypentyl) benzoate of present invention wherein
M is a divalent metal ion, such as calcium. After the reaction is
sufficiently conducted, with a same purification method with that
of preparation method 2, the compound of present invention, i.e.,
2-(.alpha.-hydroxylpentyl) benzoate wherein M is divalent metal ion
is prepared.
[0022] (5) To the solution obtained in step (2), a solution which
contains an equivalent or slightly excess amount of organic base to
the 2-(.alpha.-hydroxypentyl) benzoic acid, for instance chemically
pure aniline, is added, under the temperature of
-10.about.0.degree. C., to afford the 2-(.alpha.-hydroxypentyl)
benzoate of present invention wherein M is an organic base, such as
aniline. After the reaction is sufficiently conducted, with a same
purification method with that of preparation method 2, the compound
of present invention, i.e., 2-(.alpha.-hydroxylpentyl) benzoate
wherein M is divalent metal ion is prepared.
[0023] The acid used in the acidify reaction may be any one of
concentrated or diluted hydrochloric acid or sulfuric acid; the
temperature should be controlled within the range of
-20.about.+20.degree. C.; the organic solvent to extract
2-(.alpha.-hydroxypentyl) benzoic acid may be any one of ether,
ethyl acetate, chloroform, dichloromethane, benzene, toluene,
petroleum ether, n-hexane, or cyclohexane; the monovalent base is
selected from potassium hydroxide, sodium hydroxide, lithium
hydroxide, sodium (or potassium) methoxide, or sodium (or
potassium) ethoxide; the divalent inorganic metal salt or divalent
inorganic base is selected from MgCl.sub.2, MgCO.sub.3, CaCl.sub.2,
CaCO.sub.3, ZnCl.sub.2, ZnCO.sub.3, MgSO.sub.4, Zn(OH).sub.2,
Mg(OH).sub.2 or Ca(OH).sub.2; the organic base is selected from
aniline, benzyl amine, morpholine or diethylamine; the solvent to
solve the 2-(.alpha.-hydroxylpentyl) benzoate wherein M is a
monovalent metal salt may be any one of H.sub.2O, MeOH--H.sub.2O,
EtOH--H.sub.2O, acetone-H.sub.2O, isopropanol-H.sub.2O.
[0024] It has been found that the present compounds show good
effects on prevention and therapeutics to cardioischemia and
cerebroischemia, further, the present compounds also have
pharmacological effects of anti-platelet aggregation, arterial
occlusion of heart and brain therapeutic effects and cerebral
microcirculation amelioration effect, etc.
[0025] The present compounds have shown from animal tests excellent
effects on protecting ischemic injury of heart, anti-platelet
aggregation and alleviation of injury due to cerebral-arterial
occlusion, moreover, no side effects such as exciting or hemorrhage
have not been found.
[0026] The pharmaceutical composition of the prevent invention
comprises an treatment effective amount of the compound of the
present invention as active ingredient and a pharmaceutically
acceptable carrier.
[0027] The compounds and pharmaceutical compositions of the prevent
invention can be used for the preparation of drugs which can
prevent and treat cardioischemia and cerebroischemia, arterial
occlusion of heart and brain, amelioration of cardiac-cerebral
microcirculations.
[0028] The above mentioned "pharmaceutically acceptable carrier"
means the ordinary drug carriers such as diluents, excipients,
fillers such as starch, saccharide; binders such as cellulose
derivatives, alginates, gelatin, and polyvinylpyrrolidone;
moisturizing agents such as glycerol; disintegrating agents such as
agar, calcium carbonate and sodium bicarbonate; resorption
accelerators such as quaternary ammonium compounds; surface active
agents such as cetyl alcohol; adsorptive carriers such as kaolin
and bentonite; lubricants such as talc, calcium or magnesium
stearate, and polyethyleneglycols; further, other assistants such
as flavors and sweeteners can also be added.
[0029] The compounds of this invention can be administered orally
and intravenously to the patients in need of such treatment in a
form of pharmaceutical formulation. When administrated orally, it
can be administrated in the dosage form such as tablets, particles,
or capsules. And it can be in the form of solutions or oily/aqueous
suspensions for injection. Preferred dosage forms are tablet,
capsule and injection.
[0030] The dosage forms of present pharmaceutical compositions can
be prepared by common procedures in the art. For example, the
compounds of this invention can be admixed with one or more
carriers and formed into a desired dosage form.
[0031] The present pharmaceutical composition preferably contains
active ingredients in a weight ratio of 3:2, most preferably
1:1.
[0032] The dose of present compound may vary based on
administration route, age, weight, disease type and severeness of
disease of the patient being treated. A typical daily dose may be
from 50 mg to 600 mg per day, preferably 100.about.200 mg per day,
which can be administered once or more times.
DETAILED DESCRIPTION OF THE INVENTION
[0033] The following examples are only illustrative and are not
intended to limit the scope of the present invention.
Example 1
Preparation of Racemic Potassium 2-(.alpha.-hydroxypentyl) benzoate
(Herein after Also Referred as Potassium
dl-2-(.alpha.-hydroxypentyl) benzoate, or dl-PHPB)
[0034] Dissolving dl-3-n-Butyl-isobenzofuran-1-(3H)-one (8.5 g,
0.045 mol) in 20 ml methanol and a 10 ml methanol solution of KOH
(2.6 g, 0.046 mol) was added. The reaction solution was stirred
under reflux for one hour. After that, TLC analysis (petroleum
ether-acetone=10:1) and I.sub.2 vapor coloration showed that the
starting material was disappeared. The reaction solution was
concentrated under reduced pressure to afford a sticky yellow
residue, which was allowed to crystallize in the refrigerator after
addition of 20 ml of chloroform. The crude product was
recrystallized in MeOH--CHCl.sub.3, and a white granular crystal
(10.07 g, yield=91.50%) was obtained.
Example 2
Preparation of Potassium dl-2-(.alpha.-hydroxypentyl) benzoate
[0035] Dissolving dl-3-n-Butyl-isobenzofuran-1-(3H)-one (0.63 g,
3.3 mmol) in 10 ml methanol and a 10 ml methanol solution of KOH
(0.19 g, 3.4 mmol) was added. The reaction solution was stirred
under reflux for one hour. After that, TLC analysis (petroleum
ether-acetone=10:1) and I.sub.2 vapor coloration showed that the
starting material was disappeared. The reaction solution was
concentrated under reduced pressure to afford a sticky yellow
residue, which was allowed to crystallize in the refrigerator after
addition of 5 ml of chloroform. The crude product was
recrystallized in MeOH--CHCl.sub.3, and a white granular crystal
(0.6 g, yield=73.56%) was obtained.
Example 3
Preparation of Potassium dl-2-(.alpha.-hydroxypentyl) benzoate
[0036] Dissolving sodium dl-2-(.alpha.-hydroxypentyl) benzoate
(1.96 g, 8.5 mmol) in 10 ml of H.sub.2O and the solution was cooled
to about 0.degree. C. in an ice-salt bath. The pH was adjusted to
2.0.about.3.0 with 1N HCl and the solution was extracted quickly
with cold ether (3.times.20 ml). The ether extract was combined and
dried with anhydrous Na.sub.2SO.sub.4 at a low temperature for 3
hours and then filtered quickly under the low temperature. To the
filtrate a 20 ml methanol solution of anhydrous K.sub.2CO.sub.3
(0.58 g, 4.2 mmol) was added and the mixture was stirred fastly to
ambient temperature. A white solid appeared in the ether solution
and the solution was kept for over 24 hours, then the white solid
was filtered and dried (1.4 g, yield=66.67%).
Example 4
Preparation of Potassium dl-2-(.alpha.-hydroxypentyl) benzoate
[0037] Dissolving sodium dl-2-(.alpha.-hydroxypentyl) benzoate
(1.78 g, 7.7 mmol) in about 10 ml of H.sub.2O and the solution was
cooled to about 0.degree. C. in an ice-salt bath. The pH was
adjusted to 2.0.about.3.0 with 1N HCl and the solution was
extracted quickly with cold ether (3.times.20 ml). The ether layer
was combined and dried with anhydrous Na.sub.2SO.sub.4 at a low
temperature for 2 hours and then filtered quickly under the low
temperature. To the filtrate a 10 ml methanol solution of KOH (0.43
g, 7.7 mmol) was dropped in and the mixture was kept in the
ice-salt bath to ambient temperature. After concentration under
reduced pressure, the residue was recrystallized with MeOH-ether. A
white solid was obtained (1.3 g, yield=68.42%).
[0038] The potassium dl-2-(.alpha.-hydroxypentyl) benzoate as
prepared in the above Examples 1 till 4 procedures is a white
granular crystal.
[0039] mp. 151-152.degree. C.
[0040] IR (KBr)
[0041] 3198 cm.sup.-1 (.nu..sub.OH), 2933 cm.sup.-1 (.nu..sub.CH3),
1577, 1561 (.nu..sub.COO)
[0042] .sup.1H-NMR (300 MHz, DMSO) .delta. (ppm)
[0043] 7.65 (dd, J=6.3 Hz, 2.7 Hz, 1H), 7.17-7.05 (m, 3H), 4.32 (t,
1H), 3.40 (s, 1H), 1.80-1.55 (m, 2H), 1.38-1.04 (m, 4H), 0.81 (t,
3H)
[0044] Elemental Analysis C.sub.12H.sub.15O.sub.3K (FW246.35)
TABLE-US-00001 C(%) H(%) K(%) Calculated 58.51 6.14 15.87 Found
58.24 6.01 15.84 ##STR00002##
Example 5
Preparation of Sodium dl-2-(.alpha.-hydroxypentyl) benzoate
[0045] Dissolving dl-3-n-Butyl-isobenzofuran-1-(3H)-one (4.6 g,
0.024 mol) in 20 ml methanol and a 10 ml aqueous solution of NaOH
(0.86 g, 0.022 mol) was added. The reaction solution was stirred
under reflux for 2 hours. After that, TLC analysis (petroleum
ether-acetone=10:1) and I.sub.2 vapor coloration showed that the
starting material was disappeared. The reaction solution was
concentrated under reduced pressure to afford a sticky yellow
residue, which did not crystallize in the solvents such as
chloroform, acetone, ether or methanol, etc. A small amount of
residue was solidified with ether-ethyl acetate (10:1) to afford a
white solid, which was very hydroscopic and became sticky over
filtration. The yellow residue was washed with ether for several
times and dehydrated with anhydrous benzene, dried under reduced
pressure to afford a white foamed solid (3.67 g, yield=65.91%).
Example 6
Preparation of Sodium dl-2-(.alpha.-hydroxypentyl) benzoate
[0046] Dissolving dl-3-n-Butyl-isobenzofuran-1-(3H)-one (7.6 g,
0.04 mol) in 20 ml methanol and a fresh methanol solution (20 ml)
of sodium methoxide (0.92 g, 0.04 mol of Na) was added. The
reaction solution was stirred under reflux for 2 hours. After that,
TLC analysis (petroleum ether-acetone=10:1) and I.sub.2 vapor
coloration showed that the starting material was disappeared. The
reaction solution was concentrated under reduced pressure to afford
a sticky yellow residue, which did not crystallize in the solvents
such as chloroform, acetone, ether or methanol, etc. The yellow
residue was washed with ether for several times and dehydrated with
anhydrous benzene, dried under reduced pressure to afford a white
foamed solid (6.4 g, yield=69.56%).
Example 7
Preparation of Sodium dl-2-(.alpha.-hydroxypentyl) benzoate
[0047] Dissolving dl-PHPB (1.0 g, 0.004 mol) in 10 ml of H.sub.2O
and the solution was cooled to about 0.degree. C. in an ice-salt
bath. The pH was adjusted to 2.0.about.3.0 with 1N HCl and the
solution was extracted quickly with cold ether (3.times.20 ml). The
ether extract was combined and dried with anhydrous
Na.sub.2SO.sub.4 at a low temperature for 2 hours and then filtered
quickly under the low temperature. To the filtrate a 20 ml methanol
solution of anhydrous Na.sub.2CO.sub.3 (0.58 g, 4.2 mmol) was added
and the solution became turbid and return clear after 3 hours of
standing with ice-salt bath. The reaction solution was concentrated
under reduced pressure to afford a sticky yellow residue, which was
washed with ether for several times and dehydrated with anhydrous
benzene, dried under reduced pressure to afford a white foamed
solid (0.4 g, yield=42.78%).
Example 8
Preparation of dl-Sodium-2-(.alpha.-hydroxypentyl) benzoate
[0048] Dissolving dl-PHPB (2.4 g, 0.01 mol) in 20 ml of H.sub.2O
and the solution was cooled to about 0.degree. C. in an ice-salt
bath. The pH was adjusted to 2.0.about.3.0 with 1N HCl and the
solution was extracted quickly with cold ether (3.times.20 ml). The
ether extract was combined then a 20 ml methanol solution of NaOH
(0.39 g, 0.01 mol), the resulted solution was kept over night in
the ice-salt bath. After concentration under reduced pressure, a
sticky yellow residue was afforded, which was processed as above,
and a white foamed solid (1.2 g, yield==53.48%) was afforded.
[0049] The sodium dl-2-(.alpha.-hydroxypentyl) benzoate as prepared
in the above Examples 5 till 8 procedures is a foamed white
solid.
[0050] IR (film) 3398 cm.sup.-1 (.nu..sub.OH), 2969 cm.sup.-1
(.nu..sub.CH3), 1558, 1394 cm.sup.-1 (.nu..sub.COO)
[0051] .sup.1H-NMR (300 MHz, DMSO) .delta. (ppm) 7.70 (dd, J=6.9
Hz, 1.8 Hz, 1H), 7.17-7.07 (m, 3H), 4.39 (t, 1H), 1.65-1.48 (m,
2H), 1.39-1.10 (m, 4H), 0.83 (t, 3H)
Example 9
Preparation of Lithium dl-2-(.alpha.-hydroxypentyl) benzoate
[0052] Dissolving dl-3-n-Butyl-isobenzofuran-1-(3H)-one (1.2 g,
0.006 mol) in 15 ml methanol and LiOH.H.sub.2O (0.26 g, 0.006 mol)
was added. The reaction solution was stirred under reflux for 2
hours. After that, TLC analysis (petroleum ether-acetone=10:1) and
I.sub.2 vapor coloration showed that the starting material was
disappeared. The reaction solution was concentrated under reduced
pressure to afford a white crystal (1.1 g, yield=81.38%).
[0053] mp. 134-136.degree. C.
[0054] IR (KBr)
[0055] 3323 cm.sup.-1 (.nu..sub.OH), 2931 cm.sup.-1 (.nu..sub.CH3),
1604, 1414 cm.sup.-1 (.nu..sub.COO)
[0056] .sup.1H-NMR (300 MHz, DMSO) .delta. (ppm)
[0057] 7.66 (dd, 1H), 7.17-7.16 (m, 3H), 4.32 (t, 1H), 1.78-1.54
(m, 2H), 1.27-1.02 (m, 4H), 0.84 (t, 3H)
[0058] Elemental Analysis C.sub.12H.sub.15O.sub.3Li (FW214.19)
TABLE-US-00002 C(%) H(%) Li(%) Calculated 67.29 7.06 3.24 Found
67.34 6.87 3.26
Example 10
Preparation of Calcium dl-2-(.alpha.-hydroxypentyl) benzoate
[0059] Dissolving dl-3-n-Butyl-isobenzofuran-1-(3H)-one (1.4 g, 7.1
mmol) in 15 ml methanol and a 20 ml aqueous solution of NaOH (0.28
g, 7.0 mmol) was added. The reaction solution was stirred under
reflux for 2 hours. After that, TLC analysis (petroleum
ether-acetone=10:1) and I.sub.2 vapor coloration showed that the
starting material was disappeared. Calcium chloride (0.4 g, 4.5
mmol) was solved in 40 ml of H.sub.2O, and the solution was dropped
into the above reaction solution. The reaction was carried out in a
60.degree. C. water bath for 2 hours, and the pH was adjusted to
about 7 with 1N HCl and then reaction solution was filtered. The
filtrate was concentrated to 10 ml under reduced pressure and a
white solid appeared. The solution with the white solid appeared
was kept for 30 minutes and then filtered. The filter cake was
washed several times with water and H.sub.2O-MeOH (1:1),
respectively, until no chloride ion can be detected. Heating the
filter cake to dry to afford a white solid (1.08 g,
yield=33.23%).
Example 11
Preparation of Calcium dl-2-(.alpha.-hydroxypentyl) benzoate
[0060] Dissolving dl-3-n-Butyl-isobenzofuran-1-(3H)-one (1.1 g, 5.8
mmol) in 20 ml anhydrous ethanol and a 10 ml aqueous solution of
NaOH (0.39 g, 9.8 mmol) was added. The reaction solution was
stirred under reflux for 2 hours. After that, TLC analysis
(petroleum ether-acetone=10:1) and I.sub.2 vapor coloration showed
that the starting material was disappeared. The reaction solution
was cooled to about 0.degree. C. in an ice-salt bath, and acidified
to pH 5.0.about.6.0 with 1N HCl, additional 20 ml ethanol was added
to solve the precipitated white solid. Calcium carbonate powder
(0.33 g, 3.3 mmol) was added and the reaction solution was stirred
vigorously under low temperature and kept over night. A white solid
appeared from the reaction solution and the reaction solution was
filtered. The filter cake was washed several times with
H.sub.2O-MeOH. Heating the filter cake to dry to afford a white
solid (0.65 g, yield=24.73%).
Example 12
Preparation of Calcium dl-2-(.alpha.-hydroxypentyl) benzoate
[0061] Calcium chloride (0.12 g, 0.002 mmol) was dissolved in 20 ml
H.sub.2O, and heated to about 60.degree. C. To the solution a 10 ml
aqueous solution of dl-PHPB (0.5 g, 0.001 mol) was added. After a
while, a white solid appeared and the reaction was continued for
another 3 hours. After that, the reaction solution was filtered and
the filter cake was washed with hot water and dried under heating
to obtain a white solid (0.21 g) and the filtrate was concentrated
under reduced pressure and washed with hot water and heated to dry,
additional 0.15 g of white solid was obtained. The combined total
white solid was 0.36 g (yield=78.02%).
[0062] Calcium dl-2-(.alpha.-hydroxypentyl) benzoate as prepared in
the above Examples 9 till 12 procedures is a white solid, which
decomposes above a temperature of 252.degree. C.
[0063] IR (KBr)
[0064] 3323 cm.sup.-1 (.nu..sub.OH), 2931 cm.sup.-1 (.nu..sub.CH3),
1604, 1401 cm.sup.-1 (.nu..sub.COO)
[0065] .sup.1H-NMR (300 MHz, DMSO) .delta. (ppm)
[0066] 7.56 (d, 1H), 7.24-7.05 (m, 3H), 4.55 (t, 1H), 1.71-1.52 (m,
2H), 1.26-1.04 (m, 4H), 0.80 (t, 3H)
[0067] Elemental Analysis C.sub.24H.sub.30O.sub.6Ca (FW454.57)
TABLE-US-00003 C(%) H(%) Ca(%) Calculated 63.41 6.65 8.82 Found
63.20 6.61 9.02
Example 13
Preparation of dl-2-(.alpha.-hydroxypentyl) benzoic Acid Benzyl
Amine Salt
[0068] Dissolving dl-3-n-Butyl-isobenzofuran-1-(3H)-one (1.4 g, 7.1
mmol) in 15 ml methanol and a 20 ml aqueous solution of NaOH (0.28
g, 7.0 mmol) was added. The reaction solution was stirred
continuously under reflux for 2 hours. After that, TLC analysis
(petroleum ether-acetone=10:1) and I.sub.2 vapor coloration showed
that the starting material was disappeared. The reaction solution
was cooled to about 0.degree. C. in an ice-salt bath and acidified
with 1N HCl to pH 3.0-4.0, and the solution was extracted quickly
with cold ether (3.times.20 ml). The ether extract was combined and
dried with anhydrous Na.sub.2SO.sub.4 at a low temperature and then
filtered. Benzyl amine (0.9 ml, 8.24 mmol) was added to the
reaction solution and the reaction solution was kept over night in
the ice-salt bath. After concentration under reduced pressure, a
yellow sticky residue was obtained and a white solid (1.07 g)
separated under the addition of petroleum ether-ether, which was
recrystallized in ethyl acetate to afford a white crystal (0.61 g,
yield=26.28%).
[0069] mp. 86-88.degree. C.
[0070] IR (KBr) 3396 cm.sup.-1 (.nu..sub.NH), 2927 cm.sup.-1 (br,
.nu..sub.OH), 1637 cm.sup.-1 (.nu..sub.C.dbd.C) 1515 cm.sup.-1
(.nu..sub.COO)
[0071] .sup.1HNMR (300 MHz, DMSO) .delta. (ppm)
[0072] 7.65-7.13 (m, 9H), 4.67 (t, 1H, CH), 3.95 (d, 2H, CH.sub.2),
1.58-1.66 (m, 2H, CH.sub.2), 1.14-1.34 (m, 4H, CH.sub.2CH.sub.2),
0.82 (t, 3H, CH.sub.3)
[0073] Elemental Analysis C.sub.19H.sub.25NO.sub.3(FW315.41)
TABLE-US-00004 C(%) H(%) N(%) Calculated 72.35 7.99 4.44 Found
72.32 7.98 4.76 ##STR00003##
Example 14
Preparation of Pharmaceutical Compositions
TABLE-US-00005 [0074] Tablets Ingredient Amount (mg/tablet) Active
ingredient 50~200 Medicinal starch 20~50 Microcrystalline cellulose
15~35 Magnesium stearate 0.5 Talc 0.5~1 Sodium carboxymethyl
cellulose 2~5
[0075] The active ingredient, starch, microcrystalline cellulose
and sodium carboxymethyl cellulose were crushed and mixed. The
mixture was moisturized homogenously and powdered, which were then
sieved and dried and then sieved again. Magnesium stearate and talc
were mixed with the above mixture and the mixture was compressed to
tablets, and the tablets were coated with a film coating (which may
be hydroxypropylmethyl cellulose and the like). Each tablet
contains 50.about.200 mg of active ingredient.
Example 15
Preparation of Pharmaceutical Compositions
TABLE-US-00006 [0076] Capsules Ingredient Amount (mg/capsule)
Active ingredient 50~200 Medicinal starch or mannitose 20~50 Methyl
cellulose 3~4 Cross-linked PVP 0.5~1
[0077] The active ingredient and excipients were mixed and sieved.
The so-obtained mixture was filled into stomach-soluble hard
capsules with determine amounts. Each capsule contains 50.about.200
mg of active ingredient.
Example 16
Preparation of Pharmaceutical Compositions
TABLE-US-00007 [0078] Intravenous solution Ingredient Amount Active
ingredient 50~100 mg/bottle Sodium hydroxide Appropriate Water for
injection or saline 5~50 ml
[0079] The active ingredient was dissolved in an appropriate amount
of water for injection or isotonic saline and then filtered.
Adjusting pH to 10.5 (which may vary from 9.0 to 10.5) with
appropriate amount of NaOH. The intravenous solution was filled
into bottles under sterilizing condition.
Example 17
Preparation of Pharmaceutical Compositions
TABLE-US-00008 [0080] Lyophilized intravenous injection Ingredient
Amount Active ingredient 50~100 mg/bottle Sodium hydroxide
Appropriate
[0081] The active ingredient was dissolved in an appropriate amount
of water for injection and the pH was adjusted to 9.0.about.10.0
with NaOH. The solution was filtered and freeze-dried to afford a
cake or powder. The lyophilized injection can be injected and
transfused intravenously after it is solved in the 0.9% NaCl
solution for injection or 5% glucose injection.
Test Example 1
Effect of the Present Compounds on the Infarction Area after Local
Cerebral Ischemia in Rats
[0082] (1) Materials and Methods
[0083] Animal: male Wistar rats (250-280 g), from the Animal Center
of the Chinese Academy of Medical Sciences.
[0084] Drugs:
[0085] dl-PHPB was dissolved in double distilled water.
[0086] 2,3,5-triphenyltetrazolium chloride (TTC) was purchased from
Beijing Chemical Plant.
[0087] Methods: Middle cerebral artery occlusion (MCAO): the rats
were anesthetized by trichloroacetaldehyde monohydrate (350 mg/kg,
ip). The left CCA was exposed through a middle neck incision and
was carefully dissected free from surrounding nerves. The ICA and
ECA were isolated. Then a 4 cm long 0.26 mm diameter nylon suture
was inserted through proximal ECA into ICA for a length of 2.0 mm
from the bifurcation. The wounds were sutured and the rats were
released. The room temperature was kept 24-25.degree. C. during the
test.
[0088] Groups:
[0089] Animals were divided into two groups, 1) administration
group: dl-PHPB (200 mg/kg) and HCl (pH 1.6) 0.5 ml (to mimic the
acidic condition of the human stomach) were administrated per os
(p.o) 30 minutes prior to ischemia; 2) Control group:
double-distilled water and HCl (pH 1.6) 0.5 ml were administrated
per os (p.o) 30 minutes prior to ischemia.
[0090] Infarction Measurement:
[0091] The anaesthetized animals were decapitated 24 hours after
MCAO. Each brain were rapidly removed and kept in ice-cold saline
(0-4.degree. C.). After 10 minutes, the coronal section was sliced
into five pieces after the removal of bulbus olfactus, cerebellum,
and low brain stem. Upon cutting, the first cut was at the center
point of the connection line of the polus anterior of cerebrum and
the optic chiasm, the second cut was at the site of optic chiasm,
the third was at the site of infundibulum stalk, and the fourth was
between the infundibulum stalk and the caudal pole of posterior
lobe. The sliced brain was kept in a 5 ml solution of TTC (4%) and
K.sub.2HPO.sub.4 (IM), shaded and incubated at 37.degree. C. for 30
minutes. During the incubation, slices were turned over every 7-8
minutes. After staining, the normal cerebral tissue showed rosy
color, but the infarcted tissue showed white color. The infarcted
tissue was separated from the normal tissue and weighed. The
infarction area is calculated from the weight percentage of the
infarct tissue to the total cerebral tissue.
[0092] (2) Result:
[0093] Effect of dl-PHBP to the cerebral infarct areas in the rats
of permanent MCAO.
[0094] Table 1 shows the infarct area, the area of the dl-PHBP
treated group is 19.83.+-.3.53%, and that of the control is
26.99.+-.3.51%. The infarction is significantly reduced (P<0.01)
in the treated group compared to the control.
TABLE-US-00009 TABLE 1 Control group (%) Treated group(%) 30.50
26.01 27.08 20.24 27.92 18.24 30.64 22.74 24.87 16.84 27.44 15.93
20.46 18.79 X .+-. SD 26.99 .+-. 3.51 19.83 .+-. 3.53** **P <
0.01 vs control group
[0095] (3) Conclusion:
[0096] Potassium 2-(.alpha.-hydroxypentyl)-benzoate, dl-PHPB)
significantly reduced the cerebral tissue injury induced by MCAO
and decreased the infarction areas.
Test Example 2
The Effect of the Present Compounds on Platelet Aggregation
[0097] (1) Materials and Methods
[0098] Animal: male Wistar rats (260-280 g), from the Animal Center
of the Chinese Academy of Medical Sciences.
[0099] Drugs:
[0100] Potassium 2-(1-hydroxypentyl)-benzoate, dl-PHPB),
[0101] ADP, obtained from Shanghai Institute of Biochemistry,
Acedemia Sinica.
[0102] Equipment: Platelet Aggregometer (type: PAT-4A MEGURO-KU,
TOKYO, JAPAN.
[0103] Groups: Animals were divided into four groups:
[0104] 1) Control group: double-distilled water (400 mg/kg) was
administrated per os (p.o) 30 minutes prior blood sampling;
[0105] 2) Dl-PHPB (400 mg/kg) was administrated per os (p.o) 30
minutes prior to the blood sampling;
[0106] 3) Dl-PHPB (400 mg/kg) was administrated per os (p.o) 60
minutes prior to the blood sampling;
[0107] 4) Dl-PHPB (200 mg/kg) was administrated per os (p.o) 30
minutes prior to the blood sampling;
[0108] Method:
[0109] After oral administration of dl-PHPB, blood was drawn from
the carotid of rats at the time points of 30 and 60 minutes,
respectively. Platelet rich plasma (PRP) and platelet poor plasma
(PPP) were prepared upon common procedure. According to the method
described by of Born, PRP (200 .mu.l) was put on the Platelet
Aggregometer and pre-incubated at 37.degree. C. for 5 minutes and
then ADP with the final concentration of 5 .mu.mol/L was added to
induce platelet aggregation. The maximum aggregation was measured 5
minutes after the addition of ADP.
[0110] (2) Results
[0111] Table 2 shows the inhibitory effect on platelet aggregation
induced by ADP.
TABLE-US-00010 TABLE 2 Rate of platelet Rate of Groups N
aggregation (%) inhibition (%) Control 7 0.5556 .+-. 0.0287 --
dl-PHPB (400 mg/kg 30 min) 7 0.4483 .+-. 0.0610** 19.31 dl-PHPB
(400 mg/kg 60 min) 9 0.4989 .+-. 0.0562* 10.21 dl-PHPB (200 mg/kg
30 min) 8 0.5021 .+-. 0.0622 6.62 **P < 0.01, *P < 0.05 vs
Control group
[0112] (3) Conclusion:
[0113] Potassium 2-(1-hydroxypentyl)-benzoate, dl-PHPB)
significantly inhibits platelet aggregation compared to the control
group.
Test Example 3
The Protective Effects of dl-PHPB on Cardiac Arrhythmia Induced by
Ischemia-Reperfusion in Isolated Hearts on Rats
[0114] (1) Materials and Methods
[0115] Animals: male Wistar rats, 250-300 g, from the Animal Center
of the Chinese Academy of Medical Sciences, randomly grouped.
[0116] Drugs:
[0117] dl-PHPB,
[0118] NaCl, from the Beijing Chemical Reagent Factory No. 2,
[0119] KCl and MgSO.sub.4, Beijing Shuanghuan Chemical Reagents
Factory,
[0120] KH.sub.2PO.sub.4, Beijing Yili Fine Chemicals Co., Ltd.
[0121] NaHCO.sub.3, Beijing Chemical Reagents Company
[0122] Glucose, Beijing Guohua Chemical Reagents Factory
[0123] CaCl.sub.2, Sigma.
[0124] Apparatus:
[0125] Langendoff perfusion system;
[0126] XD-7100 ECG, Shanghai Medical Electronic Devices Factory
[0127] Methods:
[0128] (1) remove the rat heart quickly after decapitation and put
it into K--H solution at 4.degree. C., fix the aorta to the
perfusion system;
[0129] (2) perfuse the heart with K--H solution, 6-8 ml/min,
37.+-.0.5.degree. C., under the pressure of about 60 mm
H.sub.2O;
[0130] (3) connect the two copper electrodes to the cardiac apex
and the bottom of the right atria and record the cardiogram;
[0131] (4) perporate under the left anterior descending branch of
the coronary artery with 3/0 line;
[0132] (5) perfuse the heart for 10 minutes and record the normal
cardiogram;
[0133] (6) ligate the left anterior descending branch of the
coronary artery to make myocardial ischemia for 15 minutes;
[0134] (7) nip the line and resume the perfusion, record the change
of cardiogram for 30 minutes (mainly on VF and Spasmic VT);
[0135] (8) dissolve the dl-PHPB at pH 1.5 to a desired
concentration, add it into K--H solution to do the experiments.
[0136] (2) Results:
[0137] Table 3 shows the protective effects of dl-PHPB on cardiac
arrhythmia induced by ischemia-reperfusion in isolated hearts on
rats
TABLE-US-00011 TABLE 3 Spasmic Duration of VT VT VF VE Arrhythmia
Incidence Duration Incidence Incidence Incidence Group N (min) (%)
(min) (%) (%) (%) Control 12 22.5 .+-. 9.5 100 8.2 .+-. 7.9 66.7
33.3 83.3 dl-PHPB 6 23.3 .+-. 5.8 100 5.0 .+-. 6.5 50 33.3 66.7
(10.sup.-5 mol/L) dl-PHPB 7 1.7 .+-. 0.6*** 100 1.1 .+-. 0.6** 0* 0
42.9 (10.sup.-4 mol/L) VT: ventricular tachycardia VF: ventricular
fibrillation VE: ventricular ectopic beats *P < 0.05, **P <
0.01, ***P < 0.001 compared with the control group
[0138] (3) Conclusion:
[0139] Dl-PHPB can significantly shorten the duration of arrhythmia
and VT induced by ischemia-reperfusion in isolated hearts on rats,
decreases the accidence of spasmic ventricular tachycardia, and
shows significant protections on the ischemic injuries of hearts at
the concentration of 10.sup.-4 mol/L.
INDUSTRIAL AVAILABILITY
[0140] The novel 2-(.alpha.-hydroxypentyl) benzoates can be used
for the preparation of pharmaceutical compositions, which is used
for the prevention and treatment of the diseases such as cardiac
ischemia, cerebral ischemia, arterial occlusion (obstruction) of
heart and brain, etc.
* * * * *