Title: Preventive or remedy for hypertension
Abstract: Provided is a method of treating hypertension, which comprises administering an effective amount of a compound represented by the following formula (1) or (2): ##STR1##
wherein, R1 and R2 are the same or different and each independently represents hydrogen, alkyl, alkenyl, cycloalkyl, cycloalkenyl, alkoxyalkyl, aryl, alkylaryl, aralkyl, or acyl, R3 represents hydroxyl, ester bond residue, or amide bond residue, R4 represents ester bond residue or amide bond residue, or a pharmaceutically acceptable salt thereof (except ferulic acid). When the preventive or remedy for hypertension according to the present invention is administered, ferulic acid exists in the blood for a long period of time, thereby continuously suppressing a blood pressure rise. Moreover, the hypertension preventive or remedy according to the present invention has reduced in bitterness peculiar to ferulic acid, which enables patients to take it continuously.
Patent Number: 6,894,077 Issued on 05/17/2005 to Suzuki, et al.
| Inventors:
|
Suzuki; Atsushi (Haga-gun, JP);
Ochiai; Ryuji (Haga-gun, JP);
Tokimitsu; Ichiro (Haga-gun, JP)
|
| Assignee:
|
Kao Corporation (Tokyo, JP)
|
| Appl. No.:
|
161739 |
| Filed:
|
June 5, 2002 |
Foreign Application Priority Data
| Jun 05, 2001[JP] | 2001-169261 |
| Current U.S. Class: |
514/534; 514/57; 514/533; 514/561; 514/563; 514/567; 514/568; 514/730; 514/734 |
| Intern'l Class: |
A61K 031/24 |
| Field of Search: |
514/533,534,567,568,730,734,57,561,563,557
|
References Cited [Referenced By]
U.S. Patent Documents
| 6458392 | Oct., 2002 | Okawa et al.
| |
| 2002/0192317 | Dec., 2002 | Okawa et al.
| |
| Foreign Patent Documents |
| 0 387 000 | Sep., 1990 | EP.
| |
| 1 090 635 | Apr., 2001 | EP.
| |
| 1 172 112 | Jan., 2002 | EP.
| |
| 1 186 294 | Mar., 2002 | EP.
| |
| 1 186 297 | Mar., 2002 | EP.
| |
| 2 302 745 | Oct., 1976 | FR.
| |
| 2 734 478 | Nov., 1996 | FR.
| |
| WO 9101724 | Feb., 1991 | WO.
| |
| WO 9216544 | Oct., 1992 | WO.
| |
| WO 9801143 | Jan., 1998 | WO.
| |
| WO 0112178 | Feb., 2001 | WO.
| |
| WO 0112178 | Feb., 2001 | WO.
| |
| WO 0224212 | Mar., 2002 | WO.
| |
Other References
Derwent Publications, AN 1992-337587, XP-002219275, JP 04-243822, Aug. 31, 1992.
J-T. Cheng, et al., Chemical Abstracts, 1 page, XP-002219274, "Antihypertensive
Activity of Phenolics From the Flower of Lonicera japonica", 1994.
M. Carmignani, et al., Journal of Medicinal Chemistry, vol. 44, No. 18, pp. 2950-2958,
XP-002219273, "Novel Hypotensive Agents From Verbesina caracasana. 8. Synthesis
and Pharmacology of (3,4-Dimethoxycinnamoyl)-N1-Agmatine and Synthetic
Analogues", 2001.
H, P. Pham, et al., Prostaglandins in Clinical Research, pp. 609-613, XP-008007450,
"Comparative Effects on TXA2 Biosynthesis of Compounds Extracted From Three Verbenaceae
Used in African Folk Medicine", 1989.
K. Hishikawa, et al., Japanese Journal of Pharmacology, vol. 85, No. 1, p. 11P,
XP-008007448, "Possible Role of a Specific NF-K-B Inhibitor as Anti-Atherosclerosis
Agent", Mar. 21-23, 2001.
A. Suzuki, et a., Hypertension Research, vol. 25, No. 1, pp. 99-107, XP-008007437,
"Green Coffee Bean Extract and its Metabolites Have a Hypotensive Effect in Spontaneously
Hypertensive Rats", 2002.
|
Primary Examiner: Jones; Dwayne
Attorney, Agent or Firm: Oblon, Spivak, McClelland, Maier & Neustadt, P.C.
Claims
1. A method for treating hypertension, which comprises administering to a patient
in need thereof an effective amount of a composition comprising a compound of formula
(2):
##STR4##
wherein, R
1 and R
2 are the same or different and each independently
represents a hydrogen atom, an alkyl group, an alkenyl group, a cycloalkyl group,
a cycloalkenyl group, an alkoxyalkyl group, an aryl group, an alkylaryl group,
an aralkyl group or an acyl group, R
4 represents an ester bond residue
or an amide bond residue, or a pharmaceutically acceptable salt thereof.
2. The method of claim 1, wherein the compound of formula (2) is curcumin.
3. The method of claim 1, wherein the alkyl, alkenyl, cycloalkyl, cycloalkenyl,
alkoxyalkyl, aryl, alkylaryl and aralkyl groups of R
1 or R
2 are
derived from C
1-40 alcohols or aryl alcohols.
4. The method of claim 1, wherein the acyl group of R
1 or R
2
is derived from C
1-40 carboxylic acids.
5. The method of claim 1, wherein R
4 is an ester bond residue.
6. The method of claim 5, wherein the ester bond residue is selected from the
group consisting of residues derived from linear C
1-40 monohydric alcohols,
residues derived from linear C
1-40 polyhydric alcohols, residues derived
from branched C
1-40 monohydric alcohols, residues derived from branched
C
1-40 polyhydric alcohols, residues derived from hydroxyl-containing
carboxylic acids, residues derived from sugar alcohols, and residues derived from sugars.
7. The method of claim 1, wherein R
4 is an amide bond residue.
8. The method of claim 7, wherein the amide bond residue is derived from water
soluble amino acids.
9. The method of claim 1, wherein said effective amount ranges from 0.001 to
50 g.
10. The method of claim 1, wherein said composition further comprises a pharmaceutically
acceptable carrier.
11. The method of claim 1, wherein said administering is orally.
12. The method of claim 11, wherein said composition is in a form selected from
the group consisting of tablets, granules, fine subtilaes, pills, powders, hard
capsules, soft capsules, troches, chewables and liquids.
13. The method of claim 11, wherein said composition is in a liquid form.
14. The method of claim 1, wherein said administering is parenterally.
15. The method of claim 13, wherein said compound of formula (2) is in an amount
of 0.001 to 50 wt.%.
Description
BACKGROUND OF THE INVENTION
1. Field of the Invention
The present invention relates to a preventive or remedy for hypertension.
2. Description of the Related Art
Cardiac diseases such as angina pectoris, myocardial infarction and heart
failure, and cerebrovascular diseases such as cerebral infarction, cerebral hemorrhage
and subarachnoid hemorrhage have a close relation to hypertension and they are,
respectively, the second and third leading causes of death among Japanese. According
to National Livelihood Survey (fiscal 1998) of Health and Welfare Ministry, out
of 1000 patients attending a hospital, 64 patients go there for treating hypertension
and it is the first leading cause of disease in Japan. For the treatment of hypertension,
employed is drug therapy using an antihypertensive such as diuretic, sympatholytic
depressant, vasodilator or angiotensin converting enzyme inhibitor. Therapy with
such a drug is applied mainly to patients of serious hypertension. General treatment
for lifestyle modification including dietetic therapy, therapeutic exercise and
cessation of drinking or smoking is, on the other hand, employed for patients at
various stages of hypertension from mild hypertension to severe hypertension. Importance
of the general treatment has therefore been recognized recently. Of the general
treatment, improvement in eating habits is said to be important. There exists a
number of foods which have traditionally been said to have an antihypertensive
action. In addition, antihypertensive materials derived from foods have been searched
extensively, and many active ingredients having an antihypertensive action have
been separated or isolated.
Although many of the drugs employed to treat hypertension are satisfactory
in their effectiveness, they are not completely free from side effects such as
tachycardia and bradycardia and place a heavy burden on patients. Foods which are
said to have an antihypertensive action, or active ingredients thereof do not always
have satisfactory effectiveness and many of them need enough time to exhibit their
antihypertensive effect fully. Recently, the present inventor has found (in Japanese
Patent Application No. 2000-107957) that ferulic acid exhibits a high hypertension
ameliorating effect while having less side effects. It is however revealed that
the antihypertensive effect of ferulic acid does not last long because of its high
metabolic rate as measured in vivo.
An object of the present invention is therefore to provide a preventive or remedy
for hypertension which has a long lasting antihypertensive effect, has a high degree
of safety, does not impose a large stress on patients upon intake of it and has
a higher antihypertensive action.
SUMMARY OF THE INVENTION
The present inventor has found that ferulic acid has an antihypertensive action,
but owing to a high metabolic rate in vivo, its blood level reaches the maximum
about 2 hours after administration and its metabolism and excretion are completed
only after about 4 hours, while a specific compound having a ferulic acid skeleton
has a long lasting antihypertensive effect because the compound after oral administration
is metabolized into ferulic acid and this ferulic acid exists in the blood for
a long time. The present inventor has also found that a compound having the specific
ferulic acid skeleton is reduced in bitterness peculiar to ferulic acid and therefore
has an improved taste, which permits patients to take a sufficient amount daily,
and in addition it has a high degree of safety so that it is useful as a preventive
or remedy for hypertension.
In one aspect of the present invention, there is thus provided use of a compound
represented by the following formula (1) or (2):
##STR2##
wherein, R
1 and R
2 are the same or different and each
independently represents a hydrogen atom, an alkyl group, an alkenyl group, a cycloalkyl
group, a cycloalkenyl group, an alkoxyalkyl group, an aryl group, an alkylaryl
group, an aralkyl group or an acyl group, R
3 represents a hydroxyl group,
an ester bond residue or an amide bond residue, R
4 represents an ester
bond residue or an amide bond residue, or a pharmaceutically acceptable salt thereof
(except ferulic acid) for preparing a preventive or remedy for hypertension.
In another aspect of the present invention, there is also provided a method for
treating hypertension, which comprises administering an effective amount of a compound
represented by the following formula (1) or (2):
##STR3##
wherein, R
1 and R
2 are the same or different and each
independently represents a hydrogen atom, an alkyl group, an alkenyl group, a cycloalkyl
group, a cycloalkenyl group, an alkoxyalkyl group, an aryl group, an alkylaryl
group, an aralkyl group, or an acyl group, R
3 represents hydroxyl group,
an ester bond residue, or an amide bond residue, R
4 represents an ester
bond residue or an amide bond residue, or a pharmaceutically acceptable salt thereof
(except ferulic acid).
When the preventive or remedy for hypertension according to the present invention
is administered to patients, ferulic acid stays in their blood for a long period
of time, thereby continuously suppressing a blood pressure rise. Moreover, the
hypertension preventive or remedy according to the present invention has a reduced
bitterness, which bitterness is peculiar to ferulic acid, and this improved taste
enables patients to continue medication.
BRIEF DESCRIPTION OF THE INVENTION
Examples of the alkyl, alkenyl, cycloalkyl, cycloalkenyl, alkoxyalkyl, aryl,
alkylaryl and aralkyl groups in the formulas (1) and (2) include groups derived
from C
1-40 alcohols or aryl alcohols. Examples of such alcohols or aryl
alcohols include linear or branched C
1-40 alkyl or alkenyl alcohols,
aryl alcohols, monoterpene alcohols, sesquiterpene alcohols, diterpene alcohols,
triterpene alcohols, sterols, and trimethyl sterols. Specific examples include
methanol, ethanol, glycerol, oleyl alcohol, 2-ethylhexyl alcohol, allyl alcohol,
cetyl alcohol, menthyl alcohol, phenol, and benzyl alcohol. Of these groups derived
from alcohols or aryl alcohols, C
1-40 alkyl groups such as methyl, ethyl,
docosyl and tetradocosyl are preferred from the viewpoint of durability of hypotensive
effect, with ethyl, docosyl and tetradocosyl being particularly preferred.
Examples of the acyl group represented by R
1 or R
2 include
acyl groups derived from C
1-40 carboxylic acids. Such carboxylic acids
include C
1-40 carboxylic acids such as linear or branched alkyl or alkenylcarboxylic
acids, arylcarboxylic acids, monoterpenecarboxylic acids, sesquiterpenecarboxylic
acids, diterpenecarboxylic acids, triterpenecarboxylic acids and sterolcarboxylic
acids. Specific examples include formic acid, acetic acid, lactic acid, citric
acid, gluconic acid, fumaric acid, α-ketoglutaric acid, succinic acid, glycolic
acid, malic acid, tartaric acid, pyruvic acid, malonic acid, butyric acid, caproic
acid, caprylic acid, capric acid, lauric acid myristic acid, palmitic acid, stearic
acid, arachidic acid, behenic acid, valeric acid, enanthic acid, pelargonic acid,
margaric acid, myristoleic acid, palmitoleic acid, petroselinic acid, oleic acid,
vaccenic acid, linolic acid, linolenic acid, eleostearic acid, arachidonic acid,
erucic acid, glucuronic acid and mevalonic acid. Of these acyl groups, C
1-4
alkanoyl groups such as acetyl and formyl, particularly acetyl group, are
preferred from the viewpoint of the stability of the resulting compound.
The group R
3 is, as well as a hydroxyl group, a residue ester-bonded
or amide-bonded to the carboxyl group of a ferulic acid skeleton. The ester-bonded
residues include residues derived from linear or branched, monohydric or polyhydric
alcohols having 1 to 40 carbon atoms, residues derived from hydroxyl-containing
carboxylic acids, and residues derived from sugar alcohol and residues derived
from sugar. Monohydric or polyhydric alcohols include alkyl or alkenyl alcohols,
aryl alcohols, monoterpene alcohols, sesquiterpene alcohols, diterpene alcohols
and triterpene alcohols. Specific examples include monohydric alcohols such as
methanol, ethanol, propanol, butanol, pentanol, hexanol, heptanol, octanol, nonanol,
decanol, undecanol, dodecanol, tridecanol, tetradecanol, pentadecanol, hexadecanol,
heptadecanol, octadecanol, nonadecanol, eicosanol, heneicosanol, docosanol, tricosanol,
tetracosanol, oleyl alcohol, 2-ethylhexyl alcohol, allyl alcohol, cetyl alcohol,
menthyl alcohol, phenol, benzyl alcohol, and diacyl glycerol; and polyhydric alcohols
such as glycerol, monoacylglycerol and phosphatidylglycerol.
The hydroxyl-containing carboxylic acids include carboxylic acids containing
one hydroxyl group such as citric acid, isocitric acid, malic acid, glycolic acid,
cumaric acid, ferulic acid, isoferulic acid, vanillic acid, and homovanillic acid;
and carboxylic acid containing two or more hydroxyl groups such as gluconic acid,
tartaric acid, quinic acid, Shikimic acid, caffeic acid, gallic acid, vanillylmandelic
acid, glucuronic acid and mevalonic acid.
Of these hydroxyl-containing carboxylic acids, quinic acid, Shikimic acid, cinnamic
acid, cumaric acid, citric acid, caffeic acid, ferulic acid, dimethoxycinnamic
acid, gallic acid, and glucuronic acid are preferred from the viewpoint of durability
of an antihypertensive effect, with quinic acid being especially preferred.
Sugar alcohols include natural sugar alcohols, especially those contained in
plants, sugar alcohols obtained by subjecting plants to chemical treatment upon
extraction and/or fractionation, and sugar alcohols obtained by chemical modification
of natural ones. Specific examples include alcohols obtained by the reduction of
the carbonyl group of a monosaccharide, oligosaccharide or polysaccharide. Monosaccharide
alcohols include erythritol which is a four-carbon sugar alcohol obtained by fermentation
and decomposition of D-glucose with a yeast, xylitol which is a five-carbon sugar,
sorbitol which is a six-carbon sugar, and mannitol. Specific examples of oligosaccharide
include palatinit (hydrogenated palatinose), maltitol (hydrogenated maltose), lactitol
and branched oligosaccharide alcohol. Polysaccharide alcohols include hydrogenated
dextrin used as a glutinous starch syrup.
Of these sugar alcohols, erythritol, xylitol, sorbitol, and mannitol are preferred,
with erythritol being especially preferred.
Saccharides include arabinose, galactose, glucose, fructose, mannose,
ribose, maltose, cellobiose, sucrose and lactose and polymers thereof. Of these,
arabinose and galactose, and polymers thereof are especially preferred.
The amide-bonded residues as R
3 include residues derived from water
soluble amino acids. Examples of such an amino acid include glycine, alanine, valine,
leucine, isoleucine, phenylalanine, proline, serine, threonine, cysteine, cystine,
methionine, tryptophan, tyrosine, asparagine, glutamine, aspartic acid, glutamic
acid, lysine, arginine and histidine. Of these amino acids, glycine and tyrosine
are preferred from the viewpoint of durability of antihypertensive effect, with
glycine being especially preferred.
Of the groups R
3, residues derived from C
1-40 alcohols
such
as ethanol, docosanol and tetracosanol; residues derived from hydroxyl-containing
carboxylic acids such as quinic acid, gallic acid, citric acid and glucuronic acid,
and amino acid residues such as glycine are preferred from the viewpoint of the
durability of antihypertensive effect.
The group R
4 represents any one of ester-bonded residues and amide-bonded
residues, of which preferred are ester-bonded residues such as residues derived
from polyhydric alcohols such as glycerol, monoacyl glycerol and phosphatidylglycerol;
residues derived from carboxylic acid having at least 2 hydroxyl groups such as
gluconic acid, tartaric acid, quinic acid, Shikimic acid, caffeic acid, gallic
acid, vanillylmandelic acid, glucuronic acid, and mevalonic acid; and residues
derived from sugar alcohols or saccharides. When the group R
4 is a residue
derived from sugar alcohols or saccharides, the sugar alcohols or saccharides described
above as R
3 are usable as R
4.
As the group R
4, residues of arabinose, gallic acid, quinic acid and
glucuronic acid are especially preferred from the viewpoint of durability of an
antihypertensive effect.
As the compounds represented by the formula (1) or (2), preferred are chlorogenic
acids. Specific examples include 3-caffeoylquinic acid (neochlorogenic acid), 4-caffeoylquinic
acid (cryptochlorogenic acid), 5-caffeoylquinic acid (chlorogenic acid), 3,4-dicaffeoyl)
quinic acid, 3,5-dicaffeoil quinic acid, 4,5-dicaffeoyl) quinic acid, 3-feruloylquinic
acid, 4-feruloylquinic acid, 5-feruloylquinic acid, and 3-feruloyl-4-caffeoylquinic acid.
Examples of the extract from plants other than chlorogenic acids include
dimethyl caffeate ether, phenylethyl caffeate, 2-O-caffeoyl-albutin, caffeoyl-calleryanin,
3-O-caffeoyl-shikimic acid, fukinolic acid, echinacoside, 1,3-dicaffeoylquinic
acid, cichoric acid, coniferyl alcohol, curcumin, lignan and lignine.
Of these natural substances, 3-caffeoylquinic acid (neochlorogenic acid), 4-caffeoylquinic
acid (cryptochlorogenic acid), 5-caffeoylquinic acid (chlorogenic acid), 3,4-dicaffeoylquinic
acid, 3,5-dicaffeoylquinic acid, 4,5-dicaffeoylquinic acid, 3-feruloylquinic acid,
4-feruloylquinic acid, 5-feruloylquinic acid, 3-feruloyl-4-caffeoylquinic acid,
1,3-dicaffeoylquinic acid, lignan, and curcumin are preferred from the viewpoint
of durability of an antihypertensive effect, with 3-caffeoylquinic acid, 4-caffeoylquinic
acid, 5-caffeoylquinic acid and curcumin are especially preferred.
Of the compounds represented by the formula (1) or (2), especially preferred
compounds
include caffeoyl glucuronide, caffeoylglycine, feruloylglycine, feruloyl-arabinose,
3-feruloyl-4-caffeoyl arabinose, 3-caffeoylquinic acid, 4-caffeoylquinic acid and
5-caffeoylquinic acid.
The compounds of the formula (1) or (2) to be used in the invention can be extracted
from natural substances, particularly plants, containing them, while they can be
prepared industrially by chemical synthesis.
In the latter case, the compounds can be prepared by reacting ferulic acid with
an alcohol, carboxylic acid and the like corresponding to R
1, R
2,
R
3 and R
4. Ferulic acid used as the raw material can be obtained
by hydrolysis of a ferulate ester, which is available from plants, with hot sulfuric
acid under pressure, followed by purification; or by culturing bacteria (Pseudomonas)
in a broth containing a clove oil obtained by steam distillation of buds and leaves
of
Syzygium aromaticum MERRILL et PERRY or a broth containing eugenol available
by purification of the clove oil, followed by separation of the resulting culture
broth and purification. When ferulic acid is prepared by chemical synthesis, condensation
reaction of vanillin with malonic acid can be employed (Journal of American Chemical
Society, 74, 5346(1952)). Ferulic acid has stereoisomers. Any one of them is usable.
A mixture of the isomers is also usable.
Preferred examples of the plants from which the ferulate ester, among the
compounds represented by the formula (1) or (2), is extracted include coffee, apple,
grape, onion, Japanese radish, lemon,
Cnidium officinale, Angelicae radix, turpentine
tree,
Coptis Rhizome, turmeric,
Ferula assafoetida L., sweet potato,
leaves of sunflower, seeds of sunflower, jew's mallow sugarcane, corn, wheat, barley
and rice, with rice being particularly preferred. The term "rice" as used herein
means raw or dry seeds of rice (
Oryza sativa LINNE).
The compounds represented by the formula (1) or (2) include those prepared by
chemical treatment of the extract or fraction, which has been obtained from a natural
substance, particularly, a plant; and those prepared by chemical modification of
the natural substance. For example, a rice bran oil obtained from rice bran is
separated using hydrous ethanol and hexane and then ethyl ferulate is available
from the hydrous ethanol fraction.
The compounds represented by the formula (1) or (2) are available by direct extraction
from natural substances. In this case, they are prepared as a mixture of an ester
compound, amide compound and ether compound. Extraction from plants such as raw
coffee beans, leaves of a nandina and unripe apple fruits yields a mixture of chlorogenic
acids. Alternatively, extraction of the seeds of
Coffea arabica LINNE with
a warm aqueous solution of ascorbic acid or citric acid produces chlorogenic acids
to which ascorbate or citrate ester has been ester-bonded.
The compounds represented by the formula (1) or (2) have improved water solubility
and enhanced physiological availability when they are in the form of a pharmaceutically
acceptable salt. No particular limitation is imposed on the salt of ferulic acid
insofar as it is pharmaceutically acceptable. Examples of a basic substance for
the formation of such a salt include hydroxides of an alkali metal such as lithium
hydroxide, sodium hydroxide and potassium hydroxide, hydroxides of an alkaline
earth metal such as magnesium hydroxide and calcium hydroxide, inorganic bases
such as ammonium hydroxide and basic amino acids such as arginine, lysine, histidine
and ornithine, and organic bases such as monoethanolamine, diethanolamine and triethanolamine.
Of them, hydroxides of an alkali metal or alkaline earth metal are particularly preferred.
The hypertension preventive or remedy according to the present invention may
be prepared by first preparing a salt of the above-described compound and adding
the salt into a composition made of other components, or by adding ferulic acid
and a salt-forming component therewith to the composition separately, thereby forming
the salt in the resulting mixture.
The hypertension preventive or remedy according to the present invention is preferably
administered to an adult (weight: 60 kg) in an amount of about 0.001 to 50 g, preferably
about 0.003 to 20 g, especially about 0.05 to 10 g a day in terms of ferulic acid.
When a plant extract is employed, the amount in terms of dry weight can be administered.
The hypertension preventive or remedy according to the present invention can
be prepared as an orally administrable or parenterally administrable composition
by adding to its effective ingredient a pharmaceutically acceptable carrier. Of
them, the orally administrable composition is preferred. Examples of the orally
administrable composition include tablets, granules, fine subtilaes, pills, powders,
capsules (including hard capsules and soft capsules), troches, chewables and liquids
(medical drinks).
The hypertension preventive or remedy according to the present invention has
a high degree of safety so that no problem occurs even if those who have a normal
blood pressure usually take it as a food or beverage. The preventive or remedy
of the present invention can be taken as a beverage such as juice or coffee, a
liquid food such as soup, an emulsion or pasty food such as milk or curry, a semi-solid
food such as jelly and gummy, a solid food such as gum, tofu or supplement, a powdery
food, or an oil- or fat-containing food such as margarine, mayonnaise or dressing.
The compound of the present invention is added to such a beverage or drink in
an amount of 0.001 to 50 wt. %, preferably 0.01 to 25 wt. %, especially 0.1 to
10 wt. %. The content of ferulic acid is confirmed by high-performance liquid chromatography
equipped with an electrochemical detector.
EXAMPLES
Example 1
Identification of an Antihypertensive Component
1) Animals provided for test
Each of 15 week-old, spontaneously hypertensive male rats ("SHR") was anesthetized
and its blood pressure was measured at the carotid artery by using a commercially
available noninvasive sphygmomanometer for rats (manufactured by Softlon Co., Ltd.).
Its electrocardiograph was recorded by an electrocardiogram. A sample was injected
to the femoral vein through a catheter. After the rats were accustomed sufficiently
to the sphygmomanometric operation, the evaluation test was started. The rats were
all bred under conditions (in a breeding room in a rat region) at a room temperature
of 25±1° C., humidity of 55±10% RH and illumination for 12 hours
(from 7:00 am to 7:00 pm).
(2) Administration method and amount
In the control plot, physiological saline was employed. In Test plot 1, Test
plot
2 and Test plot 3, a solution of 5 μg mol/kg caffeic acid in physiological
saline, a solution of 5 μg mol/kg quinic acid in physiological saline and
a solution of 5 μg mol/kg ferulic acid in physiological saline were used, respectively.
(3) Test method
Through a catheter, the sample was intravenously administered and while administration,
systolic blood pressures of the carotid artery were measured with the passage of time.
Fluctuations in the blood pressure was not recognized when caffeic acid
or quinic acid was intravenously administered, while lowering in the blood pressure
was recognized when ferulic acid was administered.
Example 2
Measurement of an Antihypertensive Effect
1) Animals provided for test
After each of 15 week-old, spontaneously hypertensive male rats ("SHR") was
accustomed to sphygmomanometric operation by preliminarily measuring its blood
pressure for 7 successive days using a commercially available noninvasive sphygmomanometer
for rats (manufactured by Softlon Co., Ltd.), the evaluation test was started.
The rats were all bred under conditions (in a breeding room in a rat region) at
a room temperature of 25±1° C., humidity of 55±10% RH and illumination
for 12 hours (from 7:00 am to 7:00 pm).
(2) Administration method and amount
In the test plot (Comparative Example), a solution of ferulic acid (50 mg/kg
as
a dose) in physiological saline was employed. In Test plot 2, Test plot 3, Test
plot 4, Test plot 5 and Test plot 6, a solution of chlorogenic acid (50 mg/kg as
a dosage) in physiological saline, oryzanol (50 mg/kg as a dosage), curcumin (50
mg/kg as a dosage), phenylethyl caffeate (50 mg/kg as a dosage) and rosmarinic
acid (50 mg/kg as a dosage) were used, respectively. In each test plot, physiological
saline was employed as a control. Each of the samples was orally administered.
(3) Test method
SHRs were fasted overnight and divided into groups, each consisting of 5 rats.
Systolic blood pressures of the caudal artery were measured prior to administration,
and several times during 30 minutes to 24 hours after administration
(4) Statistical treatment method
The test results thus obtained were expressed by the mean value (%) and standard
deviation (SE) of the changing ratio (%) in systolic blood pressure.
A lowering ratio of the systolic blood pressure measured during 30 minutes to
24
hours each after administration relative to the systolic blood pressure prior to
administration is shown in Table 1.
| TABLE 1 |
| Successive blood pressures after the administration of various test samples |
| Control |
Mean |
0 |
-0.5 |
-0.3 |
-2.1 |
|
0.2 |
-0.5 |
|
|
|
| |
S.E |
0 |
2.4 |
1.1 |
1.9 |
|
1.4 |
1.5 |
| Test plot 1 |
Mean |
0 |
-9.5 |
-10.3 |
-6.8 |
|
-7.9 |
-3.2 |
| |
S.E |
0 |
1.5 |
1.1 |
1.4 |
|
1 |
1.7 |
| Control |
Mean |
0 |
|
|
|
-0.2 |
|
-1.3 |
-2.4 |
-2.2 |
-1.4 |
| |
S.E |
0 |
|
|
|
1.5 |
|
1 |
1 |
1.1 |
1.8 |
| Test plot 2 |
Mean |
0 |
|
|
|
-4 |
|
-6.7 |
-7.3 |
-6.8 |
-2.9 |
| |
S.E |
0 |
|
|
|
1.7 |
|
1.8 |
3.9 |
2.3 |
2.8 |
| Control |
Mean |
0 |
|
|
|
|
|
0.3 |
|
-0.5 |
5.6 |
| |
S.E |
0 |
|
|
|
|
|
2.9 |
|
2.1 |
6.9 |
| Test plot 3 |
Mean |
0 |
|
|
|
|
|
-8.5 |
|
-9.1 |
-11.0 |
| |
S.E |
0 |
|
|
|
|
|
1.8 |
|
3.8 |
2.0 |
| Test plot 4 |
Mean |
0 |
|
|
|
|
|
-11.1 |
|
-9.0 |
-11.2 |
| |
S.E |
0 |
|
|
|
|
|
1.8 |
|
0.0 |
4.5 |
| Test plot 5 |
Mean |
0 |
|
|
|
|
|
-15.2 |
|
-3.5 |
-5.3 |
| |
S.E |
0 |
|
|
|
|
|
1.4 |
|
4.5 |
5.3 |
| Test plot 6 |
Mean |
0 |
|
|
|
|
|
-13.7 |
|
-1.3 |
-0.7 |
| |
S.E |
0 |
|
|
|
|
|
1.4 |
|
0.3 |
1.6 |
| Test plot 7 |
Mean |
0 |
|
|
|
|
|
-6.4 |
|
0.9 |
2.2 |
| |
S.E |
0 |
|
|
|
|
|
6.6 |
|
1.4 |
1.6 |
| Control |
Mean |
0 |
|
-0.5 |
-2.2 |
|
-4.2 |
| |
S.E |
0 |
|
0 |
0 |
|
0 |
| Test plot 8 |
Mean |
0 |
|
-15.2 |
-12.8 |
|
-13.8 |
| |
S.E |
0 |
|
0 |
0 |
|
0 |
| * Test plot 1 (ferulic acid 50 mg/Kg) |
| Test plot 2 (5-chlorogenic acid 50 mg/Kg) |
| Test plot 3 (4-chlorogenic acid 50 mg/Kg) |
| Test plot 4 (3-chlorogenic acid 50 mg/Kg) |
| Test plot 5 (curcumin 50 mg/Kg) |
| Test plot 6 (phenylethyl caffeic acid 50 mg/Kg) |
| Test plot 7 (rosemaric acid 50 mg/Kg) |
| Test plot 8 (oryzanol 50 mg/Kg) |
As is apparent from Table 1, rats in Test plots 2 to 6 each exhibited a long-lasting
antihypertensive effect compared with those in Test plot 1 (ferulic acid).
Example 3
Measurement the Blood Level of Ferulic Acid
(1) Animals provided for the test
Each of 15 week-old, spontaneously hypertensive male rats ("SHR") was preliminarily
bred in a similar manner to Example 2.
(2) Administration method and amount
To the SHR, 200 mg/kg of chlorogenic acid was orally administered once.
(3) Test method
SHRs were fasted overnight and then divided into groups, each consisting of
5 rats. The blood levels of chlorogenic acid, caffeic acid and ferulic acid were
measured prior to administration and several times during from 30 minutes to 24
hours, each after administration.
The blood levels of chlorogenic acid, caffeic acid and ferulic acid measured
prior to administration and several times during from 30 minutes to 24 hours, each
after administration are shown in Table 2.
| TABLE 2 |
| Successive blood levels of chlorogenic acid, caffeic acid and |
| ferulic acid after the administration of chlorogenic acid |
| Chlorogenic acid |
0 |
0 |
0 |
0 |
0 |
0 |
| Caffeic acid |
0 |
0.07 |
0.179 |
0.166 |
0.05 |
0 |
| Ferulic acid |
0 |
0.074 |
0.154 |
0.174 |
0.145 |
0 |
As is apparent from Table 2, not chlorogenic acid but caffeic acid and ferulic
acid were observed in the blood, indicating that chlorogenic acid was metabolized
into caffeic acid and ferulic acid promptly after oral administration. The time-dependent
change has revealed the occurrence of in vivo conversion from caffeic acid into
ferulic acid.
Example 4
Measurement of the Blood Level of Ferulic Acid
(1) Test subject, administration method and administration amount
A beverage mixture containing an extract of raw coffee beans (chlorogenic acid
group: 280 mg/day in terms of the amount of chlorogenic acid) was fed to 5 healthy
subjects for successive 6 weeks. From the patients who did not take breakfast,
the blood was collected 24 hours after final drinking. The blood was also collected
under similar conditions from the group (placebo group: 0 mg/kg of chlorogenic
acid administered, the group consisting of 3 subjects) free from the successive
application of the above-described beverage mixture containing an extract from
raw coffee beans.
(3) Test method
The blood levels of chlorogenic acid, caffeic acid and ferulic acid were measured
using liquid chromatography.
| TABLE 3 |
| Blood levels of caffeic acid and ferulic acid 24 hours after |
| administration of chlorogenic acid |
| (280 mg/Kg in terms of human body weight) |
| |
Placebo group |
Chlorogenic acid group |
| |
| |
Caffeic acid |
0.009 |
0.0954 |
| |
Ferulic acid |
0 |
0.1044 |
| |
The results are as shown in Table 3. In the blood, not chlorogenic acid but caffeic
acid and ferulic acid were observed, suggesting that even in oral administration
to human being, chlorogenic acid was metabolized into caffeic acid and ferulic
acid. The time-dependent change has revealed the occurrence of in vivo conversion
of caffeic acid to ferulic acid.
From Examples 1 to 4, it has been understood that exhibition of the antihypertensive
effect of chlorogenic acid, quinic acid or caffeic acid owes to the metabolism
of them into ferulic acid, but chlorogenic acid, quinic acid or caffeic acid has
a markedly long-lasting antihypertensive effect so that it is useful as an antihypertensive.
Example 5
Soft Capsules
| |
| |
Gelatin |
70.0 |
(wt. %) |
| |
Glycerin |
22.9 |
| |
Methyl paraoxybenzoate |
0.15 |
| |
Methyl paraoxybenzoate |
0.15 |
| |
Propyl paraoxybenzoate |
0.51 |
| |
Water |
6.44 |
| |
Soft capsules (oval-type, weight: 150 mg) composed of the above-described composition
were filled with 400 mg of soybean oil, 50 mg of dicaffeoyl tartaric acid and 50
mg of eicosanol caffeate in a manner known per se in the art. These capsules exhibited
a good antihypertensive action when orally administered.
Example 6
The using example as a beverage will next be described.
| |
| |
Skim milk |
3.5 |
(wt. %) |
| |
Enzyme-hydrolyzed milk casein |
3.5 |
| |
Fructose |
9.0 |
| |
Eicosylferulate ester |
0.1 |
| |
3-Feruloyl-4-caffeoylarabinose |
10.0 |
| |
Citric acid |
0.1 |
| |
Ascorbic acid |
0.1 |
| |
Flavor |
0.1 |
| |
Water |
73.6 |
| |
It has been found that the beverage made of the above-described composition had
high storage stability and had good taste.
Example 7
An application example to wheat flour products will next be described.
| |
| |
Rapeseed oil |
15 |
(g) |
| |
Corn starch |
15 |
| |
Wheat flour |
42.6 |
| |
Butter |
5 |
| |
Fructose |
14 |
| |
Caffeoylglycine |
2 |
| |
Ferulyl-citric acid |
0.4 |
| |
Table salt |
0.5 |
| |
Sodium bicarbonate |
0.5 |
| |
Water |
5 |
| |
Cookies made of the above-described composition were baked.
*
