U.S. patent application number 10/433089 was filed with the patent office on 2004-02-12 for compositions for preventing or ameliorating multiple risk factor syndromes.
Invention is credited to Kishida, Hideyuki, Kitahara, Mikio, Kitano, Mitsuaki, Mae, Tatsumasa, Nakagawa, Kaku, Tsukagawa, Misuzu.
Application Number | 20040028751 10/433089 |
Document ID | / |
Family ID | 27345416 |
Filed Date | 2004-02-12 |
United States Patent
Application |
20040028751 |
Kind Code |
A1 |
Mae, Tatsumasa ; et
al. |
February 12, 2004 |
Compositions for preventing or ameliorating multiple risk factor
syndromes
Abstract
The invention aims at a composition for preventing or
ameliorating multiple risk factor syndrome involving visceral
fat-type obesity, diabetes mellitus, hyperlipemia and hypertension,
which comprises at least one member selected from the group
consisting of a licorice hydrophobic extract, a turmeric extract, a
clove extract, and a cinnamon extract.
Inventors: |
Mae, Tatsumasa;
(Kakogawa-shi, JP) ; Tsukagawa, Misuzu;
(Akashi-shi, JP) ; Kishida, Hideyuki;
(Kakogawa-shi, JP) ; Kitano, Mitsuaki;
(Takasago-shi, JP) ; Kitahara, Mikio; (Kobe-shi,
JP) ; Nakagawa, Kaku; (Kyoto-shi, JP) |
Correspondence
Address: |
KENYON & KENYON
1500 K STREET, N.W., SUITE 700
WASHINGTON
DC
20005
US
|
Family ID: |
27345416 |
Appl. No.: |
10/433089 |
Filed: |
September 11, 2003 |
PCT Filed: |
December 12, 2001 |
PCT NO: |
PCT/JP01/10869 |
Current U.S.
Class: |
424/684 |
Current CPC
Class: |
A61K 36/9066 20130101;
A61K 8/9794 20170801; A23L 33/15 20160801; A61P 9/12 20180101; Y10S
514/909 20130101; A61P 3/10 20180101; A61K 36/54 20130101; A23L
33/175 20160801; A23K 50/40 20160501; A23L 33/105 20160801; A61K
36/61 20130101; A61P 3/04 20180101; A61P 3/06 20180101; A23L 33/10
20160801; A23L 33/16 20160801; A21D 2/36 20130101; A61K 8/9789
20170801; A23K 20/10 20160501; A61P 43/00 20180101; A61Q 19/06
20130101; A23L 27/60 20160801; A61K 36/48 20130101; A61K 36/48
20130101; A61K 2300/00 20130101; A61K 36/54 20130101; A61K 2300/00
20130101; A61K 36/61 20130101; A61K 2300/00 20130101; A61K 36/9066
20130101; A61K 2300/00 20130101 |
Class at
Publication: |
424/684 |
International
Class: |
A61K 033/06 |
Foreign Application Data
Date |
Code |
Application Number |
Dec 12, 2000 |
JP |
2000-376930 |
Apr 4, 2001 |
JP |
2001-106216 |
Jun 25, 2001 |
JP |
2001-191628 |
Claims
1. A composition for preventing or ameliorating multiple risk
factor syndrome which comprises at least one member selected from
the group consisting of a licorice hydrophobic extract, a turmeric
extract, a clove extract, and a cinnamon extract.
2. The composition according to claim 1, which comprises at least
one member selected from the group consisting of a licorice
hydrophobic extract, a clove extract, and a cinnamon extract.
3. The composition according to claim 1, which comprises a licorice
hydrophobic extract.
4. The composition according to claim 1, which comprises a turmeric
extract.
5. The composition according to claim 1, which comprises a clove
extract.
6. The composition according to claim 1, which comprises a cinnamon
extract.
7. The composition according to claim 1, which is for food/beverage
use.
8. The composition according to claim 1, which is for medical
use.
9. The composition according to claim 1, which is for human
use.
10. The composition according to claim 1, which is for domestic or
pet animal use.
11. The composition according to claim 1, which is for cosmetic or
quasi-drug use.
12. The composition according to claim 1, wherein the multiple risk
factor syndrome is at least one member selected from the group
consisting of visceral fat-type obesity, diabetes mellitus,
hyperlipemia, and hypertension.
13. The composition according to claim 1, wherein the multiple risk
factor syndrome is visceral fat-type obesity.
14. The composition according to claim 1, wherein the multiple risk
factor syndrome is diabetes mellitus.
15. The composition according to claim 1, wherein the multiple risk
factor syndrome is hyperlipemia.
16. The composition according to claim 1, wherein the multiple risk
factor syndrome is hypertension.
Description
[0001] 1. Technical Field
[0002] The present invention relates to a composition for
preventing or ameliorating multiple risk factor syndrome involving
visceral fat-type obesity, diabetes mellitus, hyperlipemia and
hypertension.
[0003] 2. Background Art
[0004] Lifestyle-related desease arising from deteriorations of
living habit such as supernutrition and under exercise is a major
social concern today. Lifestyle-related desease involves various
states of desease such as obesity, diabetes mellitus, hyperlipemia
and hypertension, and these states of desease are gathering
considerable attention as multiple risk factor syndrome that maybe
an underlying disease of arteriosclerosis. Moreover, this is a
matter of concern not only in humans but also in pet animals such
as dogs and cats.
[0005] Obesity is roughly classified into subcutaneous fat-type
obesity resulting from accumulation of subcutaneous fat and
visceral fat-type obesity resulting from accumulation of
intra-abdominal fat.
[0006] Visceral fat-type obesity is a desease which is accompanied
by hyperlipemia, glucose tolerance abnormality, and hypertension at
high rates and also characterized by strong insulin resistance so
that it is known as multiple risk factor syndrome. Moreover,
visceral fat accumulation is not only found in obese individuals
but also observed in persons within the normal body weight range
and is likely to accompany the multiple risk referred to above (BIO
Clinica, September, 2000 expanded issue, 16-55). Thus, visceral fat
accumulation triggers insulin resistance, diabetes mellitus,
hyperlipemia, hypertension and the like. Matsuzawa et al. coined
the term "visceral fat syndrome" for this disease entity
(Diabetes/Metabolism Reviews, 13, 3-13, 1997). "Syndrome X" as
proposed by G. M. Reaven (Diabetes, 37, 1595-1607, 1988), "deadly
quartet" as proposed by N. M. Kaplan (Archives of Internal
Medicine, 149, 1514-1520, 1989), and "insulin-resistant syndrome"
as proposed by R. A. DeFronzo (Diabetes Care, 14, 173-194, 1991)
signify the same disease entity.
[0007] The therapy of obesity generally consists of diet therapy
and exercise therapy, and drug therapy is indicated in serious
obesity cases. However, these therapeutic regimens are recommended
or prescribed to patients by medical institutions, and are not
directed to the so-called candidate patients in the reserve.
Moreover, appetite suppressants, inhibitors of digestion and
absorption of carbohydrate and fat as energy sources, and energy
consumption promoters can be mentioned as antiobesity agents.
However, appetite suppressants and inhibitors of digestion and
absorption do not positively lower visceral fat, and energy
consumption promoters do not specifically lower visceral fat.
[0008] Japanese Kokai Publication Hei-11-187843 discloses that
administration of an extract of Cassia nomame having fat absorption
inhibitory action and an extract of Morus bombycis having
carbohydrate absorption inhibitory action in combination leads to
inhibition of body weight gain and reductions in subcutaneous fat
and visceral fat. Regarding other substances of the natural origin,
it is known that substances derived from highly unsaturated oils
such as fish oil and vegetable oil, e.g. linseed oil and perilla
oil (Japanese Kokai Publication Hei-10-231495), or conjugated
isomerized highly unsaturated fatty acids (Japanese Kokai
Publication 2000-144170) have an accumulated visceral fat lowering
function, that an amylase inhibitor derived from wheat (Japanese
Kokai Publication Hei-09-194390) inhibits visceral fat
accumulation, and that D-xylose and/or L-arabinose (Japanese Kokai
Publication Hei-07-309765 and Japanese Kokai Publication
Hei-07-242551) inhibits accumulation of body fat (subcutaneous fat
and visceral fat).
[0009] Diabetes mellitus is a disease the chief manifestation of
which is chronic hyperglycemia resulting from a deficiency in
insulin activity, and not less than 90% of diabetics are those with
type II diabetes (non-insulin dependent diabetes mellitus; NIDDM).
The number of diabetic patients in Japan has increased dramatically
from 1.57 million in 1993 to 2.18 million in 1996 (a patient survey
of the Health and Welfare Ministry). Furthermore, according to the
Health and Welfare Ministry's diabetes census undertaken in
November 1997, the persons strongly suspected to be diabetic
(inclusive of those on therapy) numbered 6.9 million and the sum of
this number and the number of individuals in whom diabetes could
not be ruled out was estimated at 13.7 million. Thus, besides the
sharply increasing number of diabetics, the number of individuals
in the reserve and not on therapy by medical institutions is by far
large, and this is a matter of serious concern.
[0010] The therapy of diabetes mellitus includes diet therapy and
exercise therapy, and in cases not sufficiently responsive to these
therapies, drug therapy is superimposed. The drug therapy includes
insulin therapy and administration of oral hypoglycemic drugs. As
oral hypoglycemic drugs, there can be mentioned sulfonylurea
derivatives such as tolbutamide and glibenclamide; biguanides such
as buformin and metformin; .alpha.-glucosidase inhibitors such as
acarbose and voglibose; and insulin resistance improving drugs such
as troglitazone and pioglitazone.
[0011] In diet therapy, the patient is recommended to refrain from
overeating and adhere to a legitimate calorie intake and, if one is
obese, instructed to make efforts to slim down to the standard body
weight. As foodstuffs to be used in diet therapy, assorted foods
for preparation of diets for diabetics, which are special-use foods
approved by the Health and Welfare Ministry, are known. However,
these are no more than the so-called low-calorie,
balanced-nutrition foods, and cannot be considered to have
intrinsic therapeutic efficacy for diabetes.
[0012] Furthermore, these therapies are prescribed for diabetics by
medical institutions and not directed to the so-called diabetics in
the reserve who outnumber the diabetics by a large margin. There is
accordingly a demand for a composition effective for preventing or
ameliorating diabetes mellitus in the form of a food/beverage, such
as food with health claims (food for specified health uses and food
with nutrient function claims) or health food, or a drug (inclusive
of a quasi-drug) to which access may be easily had by any one not
associated with medical institutions.
[0013] The incidence of diabetes has been on the steady increase in
domestic or pet animals, too, and development of a composition
effective for preventing or ameliorating diabetes in domestic and
pet animals is being desired.
[0014] Japanese Kokai Publication Hei-01-233217 discloses an
antidiabetic composition comprising curcumenone, the extract of
Curcuma aromatica, the family Zingiberaceae, as an active
ingredient. Japanese Patent Hei-06-192086 discloses an antidiabetic
composition comprising
[0015] (4S,5S)-(+)-germacrone-4,5-epoxide, the extract of Curcuma
aromatica, as an active ingredient. Curcuma aromatica is known to
be a crude drug but has not been cleared for use as a food
additive.
[0016] Licorice and its aqueous extract are in use as crude drugs
having analgesic/anticonvulsant and expectorant actions or as
foods. Since the chief component glycyrrhizin (glycyrrhizinic acid)
is about 200 times as sweet as sucrose, the "licorice extract"
obtainable by extracting licorice with water or an alkaline aqueous
medium is a food additive for use as a sweetener as well (Annotated
List of Additives in Available Books, page 163, Japanese Food
Additives Association, 1999). Its physiologic actions so far known
are adrenocortical electrolyte or glycocorticoid-like action,
estrogen-like action, testosteron production-inhibitory action,
antitussive action, antiinflammatory action, antiallergic action,
detoxicating action, hyperlipemia-improving action, gastric mucosal
cell cyclic-AMP concentration increasing action, experimental liver
impairment preventing or ameliorating action, antiviral action,
interferon-inducing action, anticaries action, tumor promoter
inhibitory action, cytosolic Ca.sup.2+ lowering action,
phospholipase A.sub.2 inhibitory action, LTB.sub.4 and PGE.sub.2
production inhibitory action, and platelet activating factor
production inhibitory action, among others. Furthermore, licorice
is one of the components of the traditional Chinese medicine
Byakko-ka-Ninjin-To which is prescribed for the diabetes-associated
intense dry-mouth, polyposia, and polyuria and, like gypsum,
anemarrhena, and ginseng which are also formulated, an aqueous
extract of licorice reportedly has a hypoglycemic action (I.
Kimura, et al.: Phytotheraphy Research, 13, 484-488, 1999).
[0017] The licorice residues which remain after extraction of
glycyrrhizin from licorice with water or an aqueous alkaline medium
are known to have a hepato-tonic (hepatoprotectant) action and/or
an antiinfective action (Japanese Kokai Publication Hei-09-143085),
an immunopotentiating action (Japanese Kokai Publication
Hei-05-262658), and an antiviral action (Japanese Kokai Publication
Hei-01-175942). Furthermore, the "licorice oily extract" which can
be obtained by extracting the residues of licorice after aqueous
washing with ethanol, acetone, or hexane is an antioxidant for use
as a food additive (Annotated Lists of Additives in Available
Books, page 164, Japanese Food Additives Association, 1999). The
licorice oily extract is known to have antibacterial activity
against Helicobacter pylori (Japanese Kokai Publication
Hei-10-130161, Japanese Kokai Publication Hei-08-119872). However,
it is not known that said licorice residues after extraction of
glycyrrhizin or said licorice oily extract ever has a visceral fat
lowering action, a hypoglycemic action, a lipid metabolism
improving action, or a blood pressure elevation inhibitory
action.
[0018] Incidentally, it is long known that an excessive intake or
prolonged use of licorice induces pseudoaldosteronism inclusive of
hypertension, edema, and hypokalemia. This condition is caused by
glycyrrhizin which is the main component of licorice. It is also
reported that hypertension may be elicited by glycyrrhetinic acid
which is a hydrolyzate of glycyrrhizin (H. Siguruonsdottir, et al.:
Journal of Human Hypertension, 15, 549-552, 2001). Thus, a licorice
aqueous extract which contains the hydrophilic fraction of licorice
and is composed predominantly of glycyrrhizin induces
hypertension.
[0019] As tropical plants relegated to the genus Curcuma, the
family Zingiberaceae, of Tropical Asian origin, several varieties
such as Curcuma longa, Curcuma aromatica, Curcuma zedoaria, and
Curcuma xanthorrhiza are known. Among these, Curcuma longa is
commonly called "turmeric" and known to be one of the component
spices of curry. This is not only used as a foodstuff but, because
the principal component curcumin of Curcuma longa is a yellow dye
(natural color), this plant or an extract thereof (curcuma dye) is
used as a dyestuff or a coloring agent (a food additive). As a
herbal medicine in traditional therapies such as Kampo (traditional
Chinese medicine) of China, Ayur-Veda of India, and Jamu of
Indonesia, it is long known that Curcuma longa has hemostatic,
stomachic, antibacterial, and antiinflammatory actions, and
actually this plant is still in use as a medicine. Furthermore, the
efficacy of turmeric (Curcuma longa) and its principal component
curcumin is attracting attention and has been found to have various
physiologic actions such as an antioxidant action, a cholagogue
action (a choleretic action), a visceral (liver, pancreas)
function-potentiating action, an anti-tumor effect, a lipid
metabolism improving action, and a skin lightening action.
[0020] However, it is not known that turmeric or its extract ever
has a visceral fat lowering action, a hypoglycemic action, or a
blood pressure elevation inhibitory action.
[0021] Clove is the flower-bud, leaf, or flower of Syzygium
aromaticum or Eugenia caryophyllata of the family Myrtaceae and is
known to be one of spices. By virtue of its
antibacterial/bactericidal action and an analgesic/anesthetic
action, clove has been used for many generations not only as a
breath sweetener or an antiodontalgic but also as a stomachic in
the realm of crude drugs and herbal medicines (Kampo medicine).
Furthermore, a clove extract has been applied as an antioxidant for
food additive use. However, it is not known that clove or its
extract ever has visceral fat lowering action, a hypoglycemic
action, a lipid metabolism improving action or a blood pressure
elevation inhibitory action.
[0022] Cinnamon or cassia is the bark of Cinnamomum cassia, C.
zeylanicum, or C. loureirii of the family Lauraceae. Cinnamon has
antibacterial and antioxidant actions and has long been used as one
of spices and its bark has been used in medicinal applications.
However, it is not known that cinnamon or its extract has a
visceral fat lowering action, a hypoglycemic action, a lipid
metabolism improving action, or a blood pressure elevation
inhibitory action.
SUMMARY OF THE INVENTION
[0023] The present invention has for its object to provide a
composition for preventing or ameliorating multiple risk factor
syndrome involving visceral fat-type obesity, diabetes mellitus,
hyperlipemia, and hypertension, without risks for side effects or
other safety problems.
[0024] The inventors of the present invention, postulating that the
cause of multiple risk factor syndrome is visceral fat-type
obesity, considered that multiple risk factor syndrome can be
prevented or ameliorated by lowering accumulated visceral fat.
[0025] Therefore, the inventors explored in earnest for possible
means for accomplishing the above object and found that a licorice
hydrophobic extract, a turmeric extract, a clove extract, and a
cinnamon extract each has an accumulated visceral fat lowering
action, a blood glucose elevation inhibitory action, a hypoglycemic
action, a lipid metabolism improving action and a blood pressure
elevation inhibitory action. The present invention has been
developed on the basis of the above finding.
[0026] The present invention, therefore, is directed to a
composition for preventing or ameliorating multiple risk factor
syndrome
[0027] which comprises at least one member selected from the group
consisting of a licorice hydrophobic extract, a turmeric extract, a
clove extract, and a cinnamon extract. This composition for
preventing or ameliorating multiple risk factor syndrome according
to the invention may be a composition for preventing or
ameliorating visceral fat-type obesity, a composition for
preventing or ameliorating diabetes mellitus, a composition for
preventing or ameliorating hyperlipemia, or a composition for
preventing or ameliorating hypertension.
[0028] The present invention is further directed to a method for
preventing or ameliorating multiple risk factor syndrome which
comprises administering or applying a composition containing at
least one member selected from the group consisting of a licorice
hydrophobic extract, a turmeric (Curcuma longa) extract, a clove
extract, and a cinnamon extract to a human being, a domestic
animal, or a pet animal.
[0029] Furthermore, the present invention is directed to the use of
at least one member selected from the group consisting of a
licorice hydrophobic extract, a turmeric extract, a clove extract,
and a cinnamon extract for the production of a composition for
preventing or ameliorating multiple risk factor syndrome.
DETAILED DESCRIPTION OF THE INVENTION
[0030] The present invention is now described in detail.
[0031] The composition of the present invention comprises at least
one member selected from the group consisting of a licorice
hydrophobic extract, a turmeric extract, a clove extract, and a
cinnamon extract. These extracts are food items and, moreover, have
been cleared for use as food additives, thus being free from side
effects and other safety problems.
[0032] The composition of the invention has a visceral fat lowering
action and, as such, is capable of preventing or ameliorating
multiple risk factor syndrome, hence, visceral fat-type obesity,
diabetes mellitus, hyperlipemia, and hypertension. Thus, in
addition to said visceral fat lowering action, it has a blood
glucose elevation inhibitory action, a hypoglycemic action, a lipid
metabolism improving action, and a blood pressure elevation
inhibitory action. The lipid metabolism improving action mentioned
just above is a total cholesterol lowering action, a triglyceride
lowering action, or a free fatty acid lowering action in serum.
[0033] The diabetes mellitus in multiple risk factor syndrome is
type II diabetes (non-insulin dependent diabetes mellitus)
resulting from glucose tolerance abnormality. Moreover, the
hyperlipemia in multiple risk factor syndrome is a state of desease
characterized by an elevation of serum total cholesterol,
triglyceride, or free fatty acid resulting from an abnormality of
lipid metabolism.
[0034] The visceral fat lowering effect can be titrated by weighing
the accumulated intra-abdominal fat, for example the fat around the
genital organ (epididymis, uterus), perirenal fat, mesenteric fat,
omental fat, etc. in experimental animals. The experimental animals
for use in this procedure are preferably dietary obesity model
animals reared on a high-fat diet (M. Rebuffe-Scrive, et al.,
Metabolism, 42, 1405-1409, 1993) or hereditary obesity model
animals, such as KK mice, KK-Ay mice, ob/ob mice, db/db mice,
Zucker fatty rats, and OLETF rats. The visceral fat lowering effect
can be evaluated from a reduction in the visceral fat area on an
abdominal CT scan image in experimental animals (C. S. Hun, et al.:
Biochemical and Biophysical Research Communications, 259, 85-90,
1999) and humans (K. Tokunaga, et al.: International Journal of
Obesity, 7, 437-445, 1983) but more expediently from a reduction in
the girth.
[0035] Furthermore, because the composition of the invention has a
visceral fat lowering action, it is capable of preventing or
ameliorating multiple risk factor syndrome involving visceral
fat-type obesity, diabetes mellitus, hyperlipemia, and
hypertension. Thus, by lowering visceral fat, the body fat
percentage can be decreased to prevent or ameliorate obesity.
Moreover, as visceral fat is decreased, the amounts of secretion of
various physiologically active substances (adipocytokines) from
adipocytes, such as leptin, TNF-.alpha. (tumor necrosis
factor-.alpha.), free fatty acids, apoprotein E, angiotensinogen,
PAI-1 (plasminogen activator inhibitor-1), etc. are decreased.
Thus, as visceral fat is decreased, TNF-.alpha. involved in insulin
resistance is decreased and the blood glucose level is depressed by
the resulting improvement in insulin resistance and remission of
glucose tolerance impairment, thus contributing to preventing or
ameliorating diabetes mellitus. Moreover, as free fatty acids and
apoprotein E, which is associated with lipid transport, are
decreased, lipid abnormalities are ameliorated to depress the blood
lipid level, thus contributing to preventing or ameliorating
hyperlipemia. Angiotensinogen, which is involved in the regulation
of blood pressure, and PAI-1, which is involved in thrombus
formation, are also decreased and the blood pressure falls,
contributing to preventing or ameliorating hypertension or
arteriosclerosis.
[0036] Licorice, the raw material of the licorice hydrophobic
extract for use in the present invention, includes Glycyrrhiza
glabra, G. uralensis, and G. inflata, of the genus Glycyrrhiza, the
family Leguminosae. Licorice is a food eaten over centuries, and
has been used as a food additive and a crude drug as well.
[0037] The licorice hydrophobic extract for use in the invention
can be obtained by extracting the active component of licorice or
its powder with an organic solvent. An alternative production
process comprises extracting out the hydrophilic component of
licorice with water or an alkaline aqueous solution in advance and
extracting the licorice residue, either as it is or after drying,
with an organic solvent. The organic solvent for use in this
process is preferably a solvent that has been cleared for use in
the production and processing of pharmaceutical products, foods,
and food additives, thus including acetone, ethanol, gycerol, ethyl
acetate, diethyl ether, cyclohexane, butanol, propanol, propylene
glycol, hexane, and methanol, among others. Oils such as edible
oils can also be used. A mixture of two or more kinds of such
solvents and a mixture of any of them with water may also be used.
Furthermore, in order to extract out the hydrophobic fraction of
licorice using a single solvent with good efficiency, ethyl
acetate, acetone or ethanol is preferred. The extract obtained by
extracting out the hydrophobic fraction of licorice or the extract
from which the extractant solvent has been removed is herein
defined as the licorice hydrophobic extract. Moreover, unless it
contains impurities objectionable in pharmaceutical products and
foods, a crude extract or a semi-purified extract can also be used
in the present invention.
[0038] Turmeric, the raw material of the turmeric extract for use
in the present invention, is the root or the rhizome of Curcuma
longa which is a perennial plant of the genus Curcuma, the family
Zingiberaceae. Turmeric is a food eaten over centuries as the spice
"turmeric", and turmeric dye obtainable by extracting turmeric with
an organic solvent, such as ethanol, hexane, or acetone, has been
cleared for use as a food additive for coloring and ranks high in
safety.
[0039] The turmeric extract for use in the invention can be
obtained by extracting the active component of turmeric or its
powder with water or an organic solvent. The organic solvent for
use in this process is preferably a solvent that has been cleared
for use in the production and processing of pharmaceutical
products, foods and food additives, thus including, acetone,
ethanol, glycerol, ethyl acetate, diethyl ether, cyclohexane,
butanol, propanol, propylene glycol, hexane, methanol, among
others. Oils such as edible oils can also be used. A mixture of two
or more kinds of such solvents and a mixture of any of them with
water may also be used. The extract obtained by the above
extraction procedure or the extract from which the extractant
solvent has been removed is herein defined as the turmeric extract.
Moreover, unless it contains impurities objectionable in
pharmaceutical products and foods, a crude extract or a
semi-purified extract can also be used in the present
invention.
[0040] Clove, the raw material of the clove extract for use in the
present invention is the flower-bud, leaf, or flower of Syzygium
aromaticum or Eugenia caryophyllata of the family Myrtaceae. Clove
is a food eaten over centuries as a spice, and the clove extract
has been cleared for use as a food additive functioning as an
antioxidant and ranks high in safety.
[0041] The clove extract for use in the invention can be obtained
by extracting the active component of clove or its powder with
water or an organic solvent. The organic solvent for use in this
process is preferably a solvent that has been cleared for use in
the production and processing of pharmaceutical products, foods and
food additives, thus including, acetone, ethanol, glycerol, ethyl
acetate, diethyl ether, cyclohexane, butanol, propanol, propylene
glycol, hexane, methanol, among others. Oils such as edible oils
can also be used. Among these solvents, two or more species may be
used as a mixture or a mixture thereof with water may also be used.
The extract obtained by the above extraction procedure or the
extract from which the extractant solvent has been removed is
herein defined as the clove extract. Moreover, unless it contains
impurities objectionable in pharmaceutical products and foods, a
crude extract or a semi-purified extract can also be used in the
present invention.
[0042] Cinnamon, the raw material of the cinnamon extract for use
in the invention is Cinnamomum cassia, C. zeylanicum, or C.
loureirii of the genus Cinnamomum of the family Lauraceae. Cinnamon
is a food eaten over centuries as a spice and has no side effect or
other safety problem.
[0043] The cinnamon extract for use in the invention can be
obtained by extracting the active component of cinnamon or its
powder with water or an organic solvent. The organic solvent for
use in this process is preferably a solvent that has been cleared
for use in the production and processing of pharmaceutical
products, foods and food additives, thus including, acetone,
ethanol, glycerol, ethyl acetate, diethyl ether, cyclohexane,
butanol, propanol, propylene glycol, hexane, methanol, among
others. Oils such as edible oils can also be used. Among these
solvents, two or more species may be used as a mixture or a mixture
thereof with water may also be used. The extract obtained by the
above extraction procedure or the extract from which the extractant
solvent has been removed is herein defined as the cinnamon extract.
Moreover, unless it contains impurities objectionable in
pharmaceutical products and foods, a crude extract or a
semi-purified extract can also be used in the present
invention.
[0044] The composition of the invention is a composition for
preventing or ameliorating multiple risk factor syndrome and
provided that it contains at least one member of the group
consisting of said licorice hydrophobic extract, turmeric extract,
clove extract, and cinnamon extract, it is not restricted in form
and can be used as a food/beverage, such as food with health claims
(food for specified health uses and food with nutrient function
claims) or health food, or as a pharmaceutical product, a cosmetic
product, or a quasi-drug.
[0045] For use as a food/beverage, it can be directly ingested or
may be formulated into easily ingestable products, such as
capsules, tablets, granules, and the like, with the aid of a known
carrier, auxiliary agent or the like for ingestion. The amount of
each extract in such a formulated product may be 0.1 to 100 weight
%, preferably 10 to 90 weight %. Furthermore, it can be mixed into
raw materials for all kinds of food and beverage products, for
example confections such as chewing gum, chocolate, candies,
jellies, biscuits, crackers, etc.; frozen sweets such as ice cream,
ice candies, etc.; beverages such as tea, nonalcoholic beverages,
nutritional drinks, drinks for beauty, etc.; noodles such as
Japanese wheat noodles, Chinese noodles, spaghetti, instant
noodles, etc.; fish paste foods such as fish minced and steamed
(kamaboko), fish sausage (chikuwa), minced flesh (hannpen), etc.;
seasonings such as dressings, mayonnaise, sauces, etc.; oleaginous
products such as margarine, butter, salad oil, etc.; bakery
products, hams, soups, retort foods, frozen foods, and so forth. In
taking such a food or beverage composition, the recommended daily
intake for an adult human is 0.1 to 1,000 mg/kg, more preferably 1
to 100 mg/kg, on an extract content basis. Such compositions can
also be used as feeds for domestic and pet animals or as pet foods,
and the recommended daily intake in these applications is
preferably 0.1 to 1,000 mg/kg on an extract content basis.
[0046] For use as a pharmaceutical product, the dosage form is not
particularly restricted but includes capsules, tablets, granules,
injections, suppositories, and patches. Such dosage forms can be
prepared by suitably formulating pharmaceutically acceptable
material for preparation such as excipient, disintegrator,
lubricant, binder, antioxidant, coloring agent, aggregation
inhibitor, absorption promoter, solubilizing agent, stabilizer, and
so on. The daily dosage of such a preparation for adult human is
0.1 to 1,000 mg/kg, preferably 1 to 100 mg/kg, on an extract
content basis, which dosage is to be administered once a day or in
a few divided doses a day. The composition can also be used as a
pharmaceutical product for domestic and pet animals and the daily
dosage for this application is preferably 0.1 to 1,000 mg/kg on an
extract content basis.
[0047] For use as a cosmetic agent or a quasi-drug, the composition
can be used in such forms as, inter alia, ointments, liniments,
aerosols, creams, soaps, face cleansers, body cleansers, toilet
water, lotions, and bath agents.
[0048] The composition of the invention can be administered or
applied to all animals inclusive of fish, reptiles, amphibians,
feathers, and mammals. The mammalian animal referred to just above
is not particularly restricted but includes human, monkeys, dogs,
cats, bovine species, equine species, swine species, sheep, mice,
rats, and guinea pigs, among others.
BEST MODE FOR CARRYING OUT THE INVENTION
[0049] The following examples illustrate the present invention in
further detail without defining the scope of the invention.
[0050] Licorice Hydrophobic Extract
PREPARATION EXAMPLE 1
Preparation of a Licorice Hydrophobic extract-1
[0051] Using a glass vessel, 500 g of licorice powder (Kaneka Sun
Spice Co., Ltd.) was steeped in 5 volumes of ethyl acetate and
allowed to stand at room temperature, protected against light, for
1 week with occasional stirring. The mixture was filtered through
filter paper (ADVANTEC No.2) twice to remove the powder and recover
an extract solution. The extract solution was concentrated under
reduced pressure to remove the solvent, whereupon 33.91 g of a
licorice hydrophobic extract was obtained.
REFERENCE EXAMPLE 1
The Glycyrrhizin Content of the Licorice Hydrophobic Extract
[0052] The glycyrrhizin content of the licorice hydrophobic extract
prepared in Preparation Example 1 was quantitated by HPLC referring
to the method of M. A. Raggi, et al. (Boll. Chim. Farmaceutico.,
133, 704-708, 1994). As the analytical column, COSMOSIL 5C18-AR,
4.6.times.250 mm (Nacalai Tesque, Inc.) was used at 40.degree. C.
Using acetonitrile:distilled water:acetic acid (35:65:0.5, v/v) as
a mobile phase run at a flow rate of 1 ml/min., the peak was
detected at 251 nm. Using ammonium glycyrrhizinate (Wako Pure
Chemical Industries, Ltd.; food additive test grade) as a
glycyrrhizin standard, a calibration curve was constructed over the
concentration range of 1 to 20 .mu.g/ml, and an assay was carried
out.
[0053] As a result, the amount of glycyrrhizin in 1 mg/ml of the
licorice hydrophobic extract was 4 .mu.g/ml, i.e. the glycyrrhizin
content of the extract was 0.4%. Since the glycyrrhizin content is
generally not less than 2.5% in licoice powders, not less than 4.5%
in aqueous extracts, or not less than 6.0% in hot water extracts,
the glycyrrhizin content of the licorice hydrophobic extract was
found to be extremely low.
TEST EXAMPLE 1
Visceral Fat Lowering Effect
[0054] C57BL/6J mice (female, 10 weeks old) were given a high-fat,
high-carbohydrate food (product of Oriental Yeast Co.; Table 1) ad
libitum for 4 to 8 weeks to establish dietary obesity. The mice
were then divided into groups of 6 to 8 and using a normal food
(product of Oriental Yeast Co.; Table 1) as a basal diet, a
non-treatment group (control group) and a group given the diet
supplemented with the licorice hydrophobic extract obtained in
Preparation Example 1 were established. In both groups, the mice
had free access to food for 4 weeks. After overnight fasting, the
abdomen was opened under ether anesthesia and the mouse was
sacrificed by exsanguination from the abdominal aorta. Then, the
periuterine fat and perirenal fat were excised and weighed. The sum
of periuterine fat weight and perirenal fat weight was recorded as
the intra-abdominal fat mass. The data are presented in Table
2.
1 TABLE 1 High-fat/ high-carbohydrate Normal food food (Modified
AIN-93G) Percent Fat 53% 22% breakdown Carbohydrate 27% 58.5%
Protein 20% 19.5% Total energy 5.100 kcal/kg 4.100 kcal/kg Formula
Casein 25.000% 20.000% Cornstarch 14.869% 49.948% Sucrose 20.000%
10.000% Soybean oil 2.000% 10.000% Lard 14.000% 0.000% Beef tallow
14.000% 0.000% Cellulose powder 5.000% 5.000% AIN-93 mineral mix
3.500% 3.500% AIN-93 vitamin mix 1.000% 1.000% Choline bitartrate
0.250% 0.250% tert-Butylhydroquinone 0.006% 0.002% L-cystine 0.375%
0.300%
[0055]
2TABLE 2 Body weight Intra-abdominal Food consumption after feeding
fat mass/body (n = 6/group, Mean .+-. SD) (g/day/mouse) (g) weight
(%) Non-treatment group 2.92 .+-. 0.39 23.0 .+-. 1.1 1.08 .+-. 0.24
(Control group) 0.1% Licorice 2.81 .+-. 0.50 22.2 .+-. 0.5 0.82
.+-. 0.15 hydrophobic extract group
[0056] It is apparent from Table 2 that compared with the
non-treatment group (control group), the group given the diet
supplemented with the licorice hydrophobic extract showed a
remarkably smaller intra-abdominal fat mass, although no difference
was found in food consumption or in body weight. It is, therefore,
clear that the visceral fat accumulated by the intake of a
high-fat, high-carbohydrate food was decreased by the intake of a
food containing the licorice hydrophobic extract.
TEST EXAMPLE 2
Preventing Effect on Diabetes Mellitus in Type II Diabetic Model
Mice-1
[0057] Using KK-Ay mice which are model animals presenting with
hereditary obesity and developing type II diabetes, the
diabetes-preventing effect of the licorice hydrophobic extract
prepared in Preparation Example 1 was evaluated. As a positive
control, the antidiabetic agent troglitazone was used.
[0058] The KK-Ay mice (female, 6 weeks old) were divided into 3
groups (5 per group), and using a normal food (Oriental Yeast Co.;
Table 1) as a basal diet, a non-treatment group (control group), a
troglitazone group, and a licorice hydrophobic extract group were
established. In all groups, the mice had free access to food for 4
weeks. Regarding troglitazone, Noscal Tab. 200 (200 mg of
troglitazone in each tablet; Sankyo Co., Ltd.) was pulverized in an
agate mortar and added to the normal food at a final concentration
of 0.2%. As the licorice hydrophobic extract, the extract prepared
in Preparation Example 1 was added to the normal food at a final
concentration of 0.15%.
[0059] During the feeding period, small samples of blood were taken
weekly from the mouse tail vein and the blood glucose was
determined with the simple blood glucose test Novo Assist Plus
(Novo Nordisk Pharma Ltd.).
[0060] At the end of the feeding period, the mice were fasted
overnight and the abdomen was opened under ether anesthesia. The
blood was withdrawn from the abdominal aorta and the liver was
excised and weighed. In addition, Sogo Ikagaku Kenkyusho Co., Ltd.
was entrusted with analyses for serum total cholesterol (T-CHO),
triglyceride (TG), free fatty acid (NEFA), GOT, GPT, LAP,
cholinesterase (ChoE), total protein (TP-S), albumin (ALB-S), and
A/G ratio.
3 TABLE 3 Mouse body weight (g) Non-treatment Licorice group
Troglitazone hydrophobic (Control group) group extract group Start
27.3 .+-. 1.0 28.0 .+-. 0.8 27.7 .+-. 0.9 1 week 33.7 .+-. 1.3 34.5
.+-. 1.9 32.9 .+-. 0.7 2 week 39.2 .+-. 0.9 40.7 .+-. 3.5 39.4 .+-.
1.7 3 week 42.1 .+-. 1.5 43.0 .+-. 5.3 41.9 .+-. 2.1 4 week 46.0
.+-. 1.6 48.6 .+-. 1.6* 43.9 .+-. 1.7 *(p < 0.05)
[0061] Mouse body weights are shown in Table 3. The mean body
weight of mice in the troglitazone group was increased slightly
over the non-treatment group (control group) and significantly
after 4 weeks. The licorice hydrophobic extract group showed no
significant difference from the non-treatment group (control
group).
4 TABLE 4 Blood glucose (mg/dl) Non-treatment Licorice group
Troglitazone hydrophobic (Control group) group extract group Start
151 .+-. 12 145 .+-. 11 157 .+-. 18 1 week 391 .+-. 95 217 .+-.
74.sunburst..sunburst. 358 .+-. 59 2 week 417 .+-. 71 173 .+-.
40.sunburst..sunburst. 290 .+-. 78.sunburst. 3 week 471 .+-. 43 304
.+-. 71.sunburst..sunburst. 342 .+-. 12.sunburst..sunburst. 4 week
441 .+-. 67 246 .+-. 51.sunburst..sunburst. 272 .+-.
48.sunburst..sunburst. .sunburst.(p < 0.05,
.sunburst..sunburst.(p < 0.01
[0062] The blood glucose data are presented in Table 4. The blood
glucose level at the start of feeding was 145 to 157 mg/dl; none of
the groups were hyperglycemic. After 1 week, the blood glucose in
the non-treatment group (control group) exceeded 390 mg/dl,
indicating the onset of diabetes. In contrast to this non-treatment
group (control group), the troglitazone group showed a significant
inhibition of blood glucose elevation, indicating that the drug
inhibits the elevation of blood glucose. The dosage of troglitazone
at this time-point as calculated from mouse body weight and food
consumption was about 270 mg/kg/day. On the other hand, the
licorice hydrophobic extract group showed a significant inhibition
of blood glucose elevation after 2 weeks, indicating that the
extract inhibits the elevation of blood glucose. The dosage of the
licorice hydrophobic extract at this time-point as calculated from
mouse body weight and food consumption was about 190 mg/kg/day.
5 TABLE 5 Non-treatment Licorice group Troglitazone hydrophobic
(Control group) group extract group Liver weight 4.23 .+-. 0.27
6.84 .+-. 1.30** 3.26 .+-. 0.51 (g/100 g body weight) T-CHO 161
.+-. 12 163 .+-. 7 131 .+-. 13** (mg/dl) TG 127 .+-. 57 39 .+-. 14*
82 .+-. 26 (mg/dl) NEFA 3299 .+-. 600 1750 .+-. 302** 2285 .+-.
175* (.mu.Eq/L) GOT 107 .+-. 22 121 .+-. 26 83 .+-. 17 (IU/L) GPT
37.4 .+-. 13.7 70.5 .+-. 26.2* 28.2 .+-. 6.3 (IU/L) LAP 63.4 .+-.
8.4 60.8 .+-. 6.1 57.4 .+-. 5.7 (IU/L) ChoE 145 .+-. 13 153 .+-. 9
145 .+-. 10 (IU/L) TP-S 5.00 .+-. 0.21 5.13 .+-. 0.17 4.90 .+-.
0.10 (g/dl) ALB-S 2.56 .+-. 0.13 2.50 .+-. 0.12 2.40 .+-. 0.07
(g/dl) A/G ratio 1.04 .+-. 0.05 0.98 .+-. 0.05 0.98 .+-. 0.08 *(p
< 0.05), **(p < 0.01)
[0063] The liver weight data and blood examination data are
presented in Table 5. Compared with the non-treatment group
(control group), the troglitazone group showed significant
decreases in TG and NEFA, indicating the lipid metabolism improving
action of the drug. However, the troglitazone group showed a
significant gain in liver weight, a significant elevation of GPT,
and a 13% elevation, which, however, is not statistically
significant, in GOT, suggesting a liver impairment. On the other
hand, the licorice hydrophobic extract group showed significant
falls in T-CHO and NEFA and a 35% decrease, which, however, is not
statistically significant, in TG, indicating the lipid metabolism
improving action of the extract. Furthermore, the licorice
hydrophobic extract group showed no significant changes in markers
of liver function such as liver weight, GOT and GPT, indicating
that the extract has no hepatotoxicity.
[0064] It was clear from the foregoing results that, like the
positive control troglitazone, the licorice hydrophobic extract has
both a blood glucose elevation inhibitory action and a lipid
metabolism improving action.
PREPARATION EXAMPLE 2
Preparation of a Licorice Hydrophobic Extract-2
[0065] Using a glass vessel, 300 g of licorice powder (Kaneka Sun
Spice Co., Ltd.) was steeped in 10 volumes of lukewarm water (about
40.degree. C.) and stirred, protected against light, overnight (for
about 18 hours). The mixture was then filtered through filter paper
(ADVANTEC No.2) and the residue obtained was dried (about
60.degree. C.) to recover 265 g of an aqueous extraction residue.
This residue was steeped in 5 volumes of ethanol and allowed to
stand at room temperature, protected against light, for 1 week with
occasional stirring. The mixture was filtered through filter paper
(ADVANTEC No.2) twice to remove the powder and recover an extract
solution. This extract solution was concentrated under reduced
pressure to remove the solvent, whereupon 23.62 g of a licorice
hydrophobic extract was obtained.
TEST EXAMPLE 3
Preventing Effect on Diabetes Mellitus in Type II Diabetic Model
Mice-2
[0066] The preventing effect on diabetes of the licorice
hydrophobic extract obtained in Preparation Example 2 was evaluated
by the same procedure as used in Test Example 2. In the licorice
hydrophobic extract group, the extract prepared in Preparation
Example 2 was added to the normal food at a final concentration of
0.2%. As a positive control, the antidiabetic agent pioglitazone
was used. As pioglitazone, Actos Tab. 30 (30 mg of pioglitazone in
each tablet; Takeda Chemical Industries, Ltd.) was pulverized in an
agate mortar and added to the normal food at a final concentration
of 0.04% as pioglitazone. For the determination of blood glucose,
the simple blood glucose test Glutest Ace (Sanwa Kagaku Kenkyusho
Co., Ltd.) was used.
6 TABLE 6 Mouse body weight (g) Non-treatment Licorice group
Pioglitazone hydrophobic (Control group) group extract group Start
27.3 .+-. 0.3 26.9 .+-. 1.0 26.4 .+-. 0.9 1 week 34.5 .+-. 0.9 36.1
.+-. 1.0 34.4 .+-. 1.9 2 week 38.9 .+-. 1.1 39.9 .+-. 1.2 38.8 .+-.
2.1 3 week 42.4 .+-. 1.5 42.9 .+-. 1.2 41.7 .+-. 1.9 4 week 43.5
.+-. 1.3 43.8 .+-. 1.4 43.2 .+-. 1.9
[0067] The mouse body weight data are presented in Table 6. The
time course of mouse body weight in the pioglitazone group and that
in the licorice hydrophobic extract group were both comparable to
the time course in the non-treatment group (control group), showing
no significant difference.
7 TABLE 7 Blood glucose (mg/dl) Non-treatment Licorice group
Pioglitazone hydrophobic (Control group) group extract group Start
142 .+-. 12 151 .+-. 9 143 .+-. 8 1 week 322 .+-. 70 163 .+-. 23**
180 .+-. 4** 2 week 427 .+-. 70 182 .+-. 9** 257 .+-. 73** 3 week
455 .+-. 66 167 .+-. 18** 206 .+-. 26** 4 week 479 .+-. 71 153 .+-.
21** 231 .+-. 59** **(p < 0.01)
[0068] The blood glucose data are presented in Table 7. In the
non-treatment group (control group), the blood glucose level was
elevated after 1 week, indicating the onset of diabetes. In
contrast, the elevation of blood glucose was significantly
inhibited in the pioglitazone group, indicating the inhibitory
action of the drug on the elevation of blood glucose. The dosage of
pioglitazone at this time-point as calculated from mouse body
weight and food consumption was about 50 mg/kg/day. The elevation
of blood glucose was significantly inhibited in the licorice
hydrophobic extract group, too, indicating the inhibitory action of
the extract on the elevation of blood glucose. The dosage of the
licorice hydrophobic extract at this time-point as calculated from
mouse body weight and food consumption was about 260 mg/kg/day.
[0069] It is clear from the foregoing results that the licorice
hydrophobic extract inhibits the elevation of blood glucose as does
the positive control pioglitazone, indicating the preventing effect
of the extract on diabetes.
TEST EXAMPLE 4
Ameliorating Effect on Diabetes Mellitus in Type II Diabetic Model
Mice
[0070] Using KK-Ay mice, the ameliorating effect on diabetes of the
licorice hydrophobic extract prepared in Preparation Example 2 was
evaluated. As a positive control, the antidiabetic agent
pioglitazone was used.
[0071] The KK-Ay mice (female, 15 weeks old) were divided into 3
groups (4 per group), and using the normal food as a basal diet, a
non-treatment group (control group), a pioglitazone group, and a
licorice hydrophobic extract group were established. The mice had
free access to food for 7 days. Pioglitazone was added to the
normal food at a final concentration of 0.02%. As the licorice
hydrophobic extract, the extract prepared in Preparation Example 2
was added to the normal food at a final concentration of 0.2%. For
the determination of blood glucose, the simple blood glucose test
Glutest Ace (Sanwa Kagaku Kenkyusho Co., Ltd.) was used.
8 TABLE 8 Blood glucose (mg/dl) Non-treatment Licorice group
Pioglitazone hydrophobic (Control group) group extract group Start
391 .+-. 45 378 .+-. 36 389 .+-. 63 Day 4 528 .+-. 52 168 .+-. 49**
192 .+-. 61** Day 7 534 .+-. 44 143 .+-. 16** 272 .+-. 61** **(p
< 0.01)
[0072] The blood glucose data are presented in Table 8. The blood
glucose level at the start of feeding was 378 to 391 mg/dl,
indicating that the mice in all groups were hyperglycemic. In the
non-treatment group (control group), the blood glucose registered
about 530 mg/dl on day 4 and day 7, indicating an exacerbation. In
contrast, in both the pioglitazone group and the licorice
hydrophobic extract group, blood glucose was decreased
significantly on both day 4 and day 7, indicating that both the
drug and the extract had a hypoglycemic action.
[0073] It was clear from the foregoing results that the licorice
hydrophobic extract has hypoglycemic action leading to an
ameliorative effect of diabetes, just as the positive control
pioglitazone.
PREPARATION EXAMPLE 3
Preparation of a Licorice Hydrophobic Extract-3
[0074] Using a stainless steel vessel, 1.5 kg of licorice powder
(Kaneka Sun Spice Co., Ltd.) was steeped in 10 volumes of water and
stirred at room temperature, protected against light, for about 24
hours. The mixture was centrifuged (5,000 g, 20 min.) and the
residue obtained was dried (about 55.degree. C.) to give an aqueous
extraction residue. This residue was steeped in 5 volumes of
ethanol and allowed to stand at room temperature, protected against
light, for 1 week with occasional stirring. The mixture was then
filtered through filter paper (ADVANTEC No. 2) twice to remove the
powder and recover an extract solution. This extract solution was
concentrated under reduced pressure to remove the solvent and
recover 141.5 g of a licorice hydrophobic extract.
TEST EXAMPLE 5
Efficacy in Spontaneously Hypertensive Rats
[0075] Using spontaneously hypertensive rats (SHR) which are model
animals of essential hypertension developing hypertension with
aging, the efficacy of the licorice hydrophobic extract prepared in
Preparation Example 3 was evaluated. As a positive control,
enalapril maleate, an antihypertensive agent, was used.
[0076] The SHRs (male, 5 week old) were divided into 3 groups (8
per group) to establish a vehicle control group, a licorice
hydrophobic extract group, and an enalapril maleate group. The
vehicle control group was treated with propylene glycol in doses of
3 ml/kg/day, the licorice hydrophobic extract group with a solution
of the licorice hydrophobic extract in propylene glycol in doses of
300 mg/3 ml/kg/day, and the enalapril maleate group with a solution
of enalapril maleate in 0.5% methylcellulose (aq. sol.) in doses of
20 mg/10 ml/kg/day, all orally for 3 consecutive weeks. During the
period, the mice had free access to food (CRF-1, Oriental Yeast
Co.) and water.
[0077] On the day immediately preceding the start of treatment and
on day 7, day 14, and day 21 of treatment, the blood pressure was
measured at the tail artery using a non-invasive automatic
hemomanometer (BP-97A, Softron Co., Ltd.). The measurement of blood
pressure was performed before each administration of the
sample.
9 TABLE 9 Body weight (g) Vehicle Licorice Enalapril Control
hydrophobic maleate group extract group group Before start 76 .+-.
2.1 76 .+-. 1.7 77 .+-. 1.7 1 week 114 .+-. 2.9 118 .+-. 1.8 115
.+-. 1.0 2 week 156 .+-. 4.1 161 .+-. 2.8 151 .+-. 1.2 3 week 192
.+-. 4.3 200 .+-. 3.1 187 .+-. 1.4
[0078] The body weights of SHRs are presented in Table 9. The time
course of body weight was almost comparable in all groups, showing
no significant difference.
10 TABLE 10 Body weight (mmHg) Vehicle Licorice Enalapril Control
hydrophobic mateate group extract group group Before start 113 .+-.
2.2 112 .+-. 2.5 111 .+-. 2.5 Day 7 137 .+-. 4.0 125 .+-.
3.3.sunburst. 118 .+-. 3.2.sunburst..sunburst. Day 14 156 .+-. 3.3
143 .+-. 5.2 119 .+-. 3.4.sunburst..sunburst. Day 21 166 .+-. 2.9
153 .+-. 2.8.sunburst..sunburst. 134 .+-. 3.1.sunburst..sunburst.
.sunburst.(p < 0.05), .sunburst..sunburst.(p < 0.01)
[0079] The blood pressure data are presented in Table 10. The blood
pressure before the start of administration was 111 to 113 mmHg;
thus no group showed a hypertensive state. In the vehicle control
group, the blood pressure began to rise on day 7, indicating the
onset of hypertension. Compared with the vehicle control group, the
elevation of blood pressure was inhibited in the licorice
hydrophobic extract group, with a significant difference being
noted on day 7 and on day 21. In the antihypertensive agent
enalapril maleate group, the elevation of blood pressure was
significantly inhibited on and after day 7.
[0080] It is clear from the foregoing results that the licorice
hydrophobic extract is effective in preventing or ameliorating
hypertension.
11 (Example 1) Preparation of licorice hydrophobic
extract-containing tablets Licorice hydrophobic extract 45 weight
parts Lactose 35 weight parts Crystalline cellulose 15 weight parts
Sucrose fatty acid ester 5 weight parts
[0081] According to the above recipe, licorice hydrophobic
extract-containing tablets for food/beverage or medical use were
manufactured by the established procedure.
12 (Example 2) Preparation of licorice hydrophobic
extract-containing soft capsules Licorice hydrophobic extract 40
weight parts Sesame oil 55 weight parts Glycerin fatty acid ester 5
weight parts
[0082] According to the above recipe, licorice hydrophobic
extract-containing soft capsules for food/beverage or medical use
were manufactured by the established procedure.
13 (Example 3) Preparation of licorice hydrophobic
extract-containing crackers Licorice hydrophobic extract 1 weight
part.sup. Soft flour 120 weight parts Common salt 1 weight
part.sup.11 Baking powder 2 weight parts Butter 30 weight parts
Water 40 weight parts
[0083] According to the above recipe, licorice hydrophobic
extract-containing crackers were manufactured by the established
procedure.
14 (Example 4) Preparation of licorice hydrophobic
extract-containing noodles Licorice hydrophobic extract 1 weight
part.sup. Hard flour 100 weight parts Soft flour 100 weight parts
Common salt 10 weight parts Water 100 weight parts
[0084] According to the above recipe, licorice hydrophobic
extract-containing noodles were manufactured by the established
procedure.
15 (Example 5) Preparation of a licorice hydrophobic
extract-containing dressing Licorice hydrophobic extract 10 weight
parts Olive oil 80 weight parts Vinegar 60 weight parts Common salt
3 weight parts Pepper 1 weight part.sup. Lemon juice 5 weight
parts
[0085] According to the above recipe, a licorice hydrophobic
extract-containing dressing was manufactured by the established
procedure.
[0086] Turmeric Extract
PREPARATION EXAMPLE 4
Preparation of a Turmeric Extract-1
[0087] Using a glass vessel, 400 g of turmeric powder (Kaneka Sun
Spice Co., Ltd.) was steeped in 5 volumes of ethanol and allowed to
stand at room temperature, protected against light, for 1 week with
occasional stirring. The mixture was filtered through filter paper
(ADVANTEC No.2) twice to remove the powder and recover an extract
solution. This extract solution was concentrated under reduced
pressure to remove the solvent and recover 43.71 g of a turmeric
extract.
TEST EXAMPLE 6
Visceral Fat Lowering Effect
[0088] C57BL/6J mice (female, 10 weeks old) were given a high-fat,
high-carbohydrate food (product of Oriental Yeast Co.; Table 1) ad
libitum for 4 to 8 weeks to establish dietary obesity. The mice
were then divided into groups of 6 to 8 and using a normal food
(product of Oriental Yeast Co.; Table 1) as a basal diet, a
non-treatment group (control group) and a group given the diet
supplemented with the turmeric extract obtained in Preparation
Example 4 were established. In both groups, the mice had free
access to food for 4 weeks. After overnight fasting, the abdomen
was opened under ether anesthesia and the mouse was sacrificed by
exsanguination from the abdominal aorta. Then, the periuterine fat
and perirenal fat were excised and weighed. The sum of periuterine
fat weight and perirenal fat weight was recorded as the
intra-abdominal fat mass. The data are presented in Table 11 and
12.
16TABLE 11 Body weight Intra-abdominal Food consumption after
feeding fat mass/body (n = 8/group, Mean .+-. SD) (g/day/mouse) (g)
weight (%) Non-treatment group 3.17 .+-. 0.43 22.9 .+-. 1.5 1.76
.+-. 0.73 (Control group) 1% Turmeric 2.92 .+-. 0.62 21.7 .+-. 1.1
0.71 .+-. 0.20 extract group
[0089]
17TABLE 12 Body weight Intra-abdominal Food consumption after
feeding fat mass/body (n = 6/group, Mean .+-. SD) (g/day/mouse) (g)
weight (%) Non-treatment group 2.92 .+-. 0.39 23.0 .+-. 1.1 1.08
.+-. 0.24 (Control group) 0.5% Turmeric 2.53 .+-. 0.59 22.6 .+-.
0.9 0.00 .+-. 0.50 extract group
[0090] It is apparent from Tables 11 and 12 that compared with the
non-treatment group (control group), the turmeric extract group
showed no difference in food consumption or in body weight but the
intra-abdominal fat mass was remarkably decreased in this group. It
was clear that the visceral fat accumulated by the intake of the
high-fat, high-carbohydrate food was decreased by the intake of the
turmeric extract-supplemented food.
PREPARATION EXAMPLE 5
Preparation of a Turmeric Extract-2
[0091] Using a glass vessel, 793 g of turmeric powder (Kaneka Sun
Spice Co., Ltd.) was steeped in 4,000 ml of ethanol and allowed to
stand at room temperature, protected against light, for 1 week with
occasional stirring. The mixture was filtered through filter paper
(ADVANTEC No.2) twice to remove the powder and recover an extract
solution. This extract solution was concentrated under reduced
pressure to remove the solvent and recover 79.48 g of a turmeric
extract.
TEST EXAMPLE 7
Efficacy in Type II Diabetic Model Mice
[0092] Using KK-Ay mice which are model animals presenting with
hereditary obesity and developing type II diabetes, the effect of
the turmeric extract was evaluated. As a positive control, the
antidiabetic agent pioglitazone was used.
[0093] The KK-Ay mice (female, 6 weeks old) were divided into 3
groups (5 per group) and using the normal food (Oriental Yeast Co.;
Table 1) as a basal diet, the animals in the non-treatment group
(control group), pioglitazone group, and turmeric extract group
were fed ad libitum for 4 weeks. As pioglitazone, Actos Tab. 30 (30
mg of pioglitazone in each tablet; Takeda Chemical Industries,
Ltd.) was pulverized in an agate mortar and added to the normal
food at a final concentration of 0.04% as pioglitazone. As to the
turmeric extract, the extract prepared in Preparation Example 5 was
added to the normal food at a final concentration of 0.5%.
[0094] During the feeding period, small samples of blood were taken
weekly from the mouse tail vein and the blood glucose was measured
with the simple blood glucose test Glutest Ace (Sanwa Kagaku
Kenkyusho Co., Ltd.).
18 TABLE 13 Mouse body weight (g) Non-treatment group Pioglitazone
Turmeric extract (Control group) group group Start 27.3 .+-. 0.3
26.9 .+-. 1.0 27.1 .+-. 0.8 1 week 34.5 .+-. 0.9 36.7 .+-. 1.0 35.3
.+-. 1.1 2 week 38.9 .+-. 1.1 39.9 .+-. 1.2 39.4 .+-. 1.2 3 week
41.0 .+-. 1.3 42.3 .+-. 1.2 41.8 .+-. 1.3 4 week 43.5 .+-. 1.3 43.8
.+-. 1.4 44.4 .+-. 1.6
[0095] The body weights of mice are presented in Table 13. Both the
pioglitazone group and the turmeric extract group followed nearly
the same time courses of body weight as the non-treatment group
(control group), showing no significant difference.
19 TABLE 14 Blood glucose (mg/dl) Non-treatment group Pioglitazone
Turmeric extract (Control group) group group Start 142 .+-. 12 151
.+-. 9 142 .+-. 6 1 week 322 .+-. 70 163 .+-. 23** 193 .+-. 28** 2
week 427 .+-. 70 182 .+-. 9** 201 .+-. 50** 3 week 455 .+-. 66 166
.+-. 18** 244 .+-. 41** 4 week 479 .+-. 71 153 .+-. 21** 270 .+-.
31** **(p < 0.01)
[0096] The blood glucose data are presented in Table 14. The blood
glucose level at the start of feeding was 142 to 151 mg/dl, with
none of the groups being hyperglycemic. In the non-treatment group
(control group), the blood glucose level was elevated after week 1,
indicating the onset of diabetes. In the turmeric extract group,
compared with the non-treatment group (control group), the
elevation of blood glucose was significantly inhibited as it was
true of the antidiabetic pioglitazone group, indicating that the
extract has a blood glucose lowering action. The dosage of the
turmeric extract at this time-point as calculated from mouse body
weight and food consumption was about 670 mg/kg/day. It is clear
from the foregoing results that the turmeric extract is effective
in preventing or ameliorating diabetes mellitus.
20 (Example 6) Preparation of turmeric extract-containing tablets
Turmeric extract 45 weight parts Lactose 35 weight parts
Crystalline cellulose 15 weight parts Sucrose fatty acid ester 5
weight parts
[0097] According to the above recipe, turmeric extract-containing
tablets for food/beverage or medical use were manufactured by the
established procedure.
21 (Example 7) Preparation of turmeric extract-containing soft
capsules Turmeric extract 40 weight parts Sesame oil 55 weight
parts Glycerin fatty acid ester 5 weight parts
[0098] According to the above recipe, turmeric extract-containing
soft capsules for food/beverage or medical use were manufactured by
the established procedure.
22 (Example 8) Preparation of turmeric extract-containing crackers
Turmeric extract 1 weight part.sup. Soft flour 120 weight parts
Common salt 1 weight part.sup. Baking powder 2 weight parts Butter
30 weight parts Water 40 weight parts
[0099] According to the above recipe, turmeric extract-containing
crackers were manufactured by the established method.
23 (Example 9) Preparation of turmeric extract-containing noodles
Turmeric extract 1 weight part.sup. Hard flour 100 weight parts
Soft flour 100 weight parts Common salt 10 weight parts Water 100
weight parts
[0100] According to the above recipe, turmeric extract-containing
noodles were manufactured by the established method.
24 (Example 10) Preparation of a turmeric extract-containing
dressing Turmeric extract 10 weight parts Olive oil 80 weight parts
Vinegar 60 weight parts Common salt 3 weight parts Pepper 1 weight
part.sup. Lemon juice 5 weight parts
[0101] According to the above recipe, a turmeric extract-containing
dressing was manufactured by the established procedure.
[0102] Clove Extract
PREPARATION EXAMPLE 6
Preparation of a Clove Extract-1
[0103] Using a glass vessel, 600 g of clove powder (Kaneka Sun
Spice Co., Ltd.) was steeped in 5 volumes of ethyl acetate and
allowed to stand at room temperature, protected against light, for
1 week with occasional stirring. The mixture was then filtered
through filter paper (ADVANTEC No. 2) twice to remove the powder
and recover an extract solution. This extract solution was
concentrated under reduced pressure to remove the solvent and
recover 47.59 g of a clove extract.
TEST EXAMPLE 8
Visceral Fat Lowering Effect
[0104] C57BL/6J mice (female, 10 weeks old) were given a high-fat,
high-carbohydrate food (product of Oriental Yeast Co.; Table 1) ad
libitum for 4 to 8 weeks to establish dietary obesity. The mice
were then divided into groups of 6 to 8 and using a normal food
(product of Oriental Yeast Co.; Table 1) as a basal diet, a
non-treatment group (control group) and a group given the diet
supplemented with the clove extract obtained in Preparation Example
6 were established. In both groups, the mice had free access to
food for 4 weeks. After overnight fasting, the abdomen was opened
under ether anesthesia and the mouse was sacrificed by
exsanguination from the abdominal aorta. Then, the periuterine fat
and perirenal fat were excised and weighed. The sum of periuterine
fat weight and perirenal fat weight was recorded as the
intra-abdominal fat mass. The data are presented in Table 15.
25TABLE 15 Body weight Intra-abdominal Food consumption after
feeding fat mass/body (n = 8/group, Mean .+-. SD) (g/day/mouse) (g)
weight (%) Non-treatment group 3.17 .+-. 0.43 22.9 .+-. 1.5 1.76
.+-. 0.73 (Control group) 1% Clove 2.55 .+-. 0.53 21.7 .+-. 0.6
0.79 .+-. 0.15 extract group
[0105] It is apparent from Table 15 that compared with the
non-treatment group (control group), the clove extract group showed
no difference in food consumption or in body weight but the
intra-abdominal fat mass was remarkably decreased in this group. It
was clear that the visceral fat accumulated by the intake of the
high-fat, high-carbohydrate food was decreased by the intake of the
clove extract-supplemented food.
PREPARATION EXAMPLE 7
Preparation of a Clove Extract-2
[0106] Using a glass vessel, 2 kg of clove powder (Kaneka Sun Spice
Co., Ltd.) was steeped in 10 L of ethanol and allowed to stand at
room temperature, protected against light, for 1 week with
occasional stirring. The mixture was filtered through filter paper
(ADVANTEC No. 2) twice to remove the powder and recover an extract
solution. This extract solution was concentrated under reduced
pressure to remove the solvent and recover 217 g of a clove
extract.
TEST EXAMPLE 9
Efficacy in Type II Diabetic Model Mice
[0107] Using KK-Ay mice which are model animals presenting with
hereditary obesity and developing type II diabetes, the effect of
the clove extract was evaluated. As a positive control, the
antidiabetic agent pioglitazone was used.
[0108] The KK-Ay mice (female, 6 weeks old) were divided into 3
groups (5 per group) and using the normal food (Oriental Yeast Co.;
Table 1) as a basal diet, the animals in the non-treatment group
(control group), pioglitazone group, and clove extract group were
fed ad libitum for 3 weeks. As pioglitazone, Actos Tab. 30 (30 mg
of pioglitazone in each tablet; Takeda Chemical Industries, Ltd.)
was pulverized in an agate mortar and added to the normal food at a
final concentration of 0.02% as pioglitazone. As to the clove
extract, the extract prepared in Preparation Example 7 was added to
the normal food at a final concentration of 0.5%.
[0109] During the feeding period, small samples of blood were taken
weekly from the mouse tail vein and the blood glucose was measured
with the simple blood glucose test Glutest Ace (Sanwa Kagaku
Kenkyusho Co., Ltd.).
26 TABLE 16 Mouse body weight (g) Non-treatment group Pioglitazone
Clove extract (Control group) group group Start 26.7 .+-. 1.1 27.4
.+-. 0.5 27.1 .+-. 1.1 1 week 34.2 .+-. 2.1 35.0 .+-. 1.1 34.7 .+-.
1.9 2 week 39.2 .+-. 2.0 41.0 .+-. 1.5 39.2 .+-. 1.4 3 week 42.7
.+-. 2.5 45.4 .+-. 1.4 42.5 .+-. 2.1
[0110] The body weights of mice are presented in Table 16. Both the
pioglitazone group and the clove extract group followed nearly the
same time courses of body weight as the non-treatment group
(control group), showing no significant difference.
27 TABLE 17 Blood glucose (mg/dl) Non-treatment group Pioglitazone
Clove extract (Control group) group group Start 163 .+-. 14 156
.+-. 16 171 .+-. 14 1 week 304 .+-. 11 208 .+-. 21 289 .+-. 63 2
week 457 .+-. 48 192 .+-. 17** 214 .+-. 67** 3 week 514 .+-. 60 164
.+-. 16** 311 .+-. 90** *(p < 0.05), **(p < 0.01)
[0111] The blood glucose data are presented in Table 17. The blood
glucose level at the start of feeding was 156 to 171 mg/dl, with
none of the groups being hyperglycemic. In the non-treatment group
(control group), the blood glucose level rose after week 1,
indicating the onset of diabetes. In the clove extract group,
compared with the non-treatment group (control group), the
elevation of blood glucose was significantly inhibited as it was
true of the antidiabetic pioglitazone group, indicating that the
extract has a blood glucose lowering action. The dosage of the
clove extract at this time-point as calculated from mouse body
weight and food consumption was about 660 mg/kg/day.
28 (Example 11) Preparation of clove extract-containing tablets
Clove extract 45 weight parts Lactose 35 weight parts Crystalline
cellulose 15 weight parts Sucrose fatty acid ester 5 weight
parts
[0112] According to the above recipe, clove extract-containing
tablets for food/beverage or medical use were manufactured by the
established procedure.
29 (Example 12) Preparation of clove extract-containing soft
capsules Clove extract 40 weight parts Sesame oil 55 weight parts
Glycerin fatty acid ester 5 weight parts
[0113] According to the above recipe, clove extract-containing soft
capsules for food/beverage or medical use were manufactured by the
established procedure.
30 (Example 13) Preparation of clove extract-containing crackers
Clove extract 1 weight part.sup. Soft flour 120 weight parts Common
salt 1 weight part.sup. Baking powder 2 weight parts Butter 30
weight parts Water 40 weight parts
[0114] According to the above recipe, clove extract-containing
crackers were manufactured by the established method.
31 (Example 14) Preparation of clove extract-containing noodles
Clove extract 1 weight part.sup. Hard flour 100 weight parts Soft
flour 100 weight parts Common salt 10 weight parts Water 100 weight
parts
[0115] According to the above recipe, clove extract-containing
noodles were manufactured by the established method.
32 (Example 15) Preparation of a clove extract-containing dressing
Clove extract 10 weight parts Olive oil 80 weight parts Vinegar 60
weight parts Common salt 3 weight parts Pepper 1 weight part.sup.
Lemon juice 5 weight parts
[0116] According to the above recipe, a clove extract-containing
dressing was manufactured by the established method.
[0117] Cinnamon Extract
PREPARATION EXAMPLE 8
Preparation of a Cinnamon Extract
[0118] Using a glass vessel, 1,000 g of cinnamon powder (Kaneka Sun
Spice Co., Ltd.) was steeped in 5 volumes of ethyl acetate and
allowed to stand at room temperature, protected against light, for
1 week with occasional stirring. The mixture was then filtered
through filter paper (ADVANTEC No. 2) twice to remove the powder
and recover an extract solution. This extract solution was
concentrated under reduced pressure to remove the solvent and
recover 59.57 g of a cinnamon extract.
TEST EXAMPLE 10
Visceral Fat Lowering Effect
[0119] C57BL/6J mice (female, 10 weeks old) were given a high-fat,
high-carbohydrate food (product of Oriental Yeast Co.; Table 1) ad
libitum for 4 to 8 weeks to establish dietary obesity. The mice
were then divided into groups of 6 to 8 and using a normal food
(product of Oriental Yeast Co.; Table 1) as a basal diet, a
non-treatment group (control group) and a group given the diet
supplemented with the cinnamon extract obtained in Preparation
Example 8 were established. In both groups, the mice had free
access to food for 4 weeks. After overnight fasting, the abdomen
was opened under ether anesthesia and the mouse was sacrificed by
exsanguination from the abdominal aorta. Then, the periuterine fat
and perirenal fat were excised and weighed. The sum of periuterine
fat weight and perirenal fat weight was recorded as the
intra-abdominal fat mass. The data are presented in Table 18.
33TABLE 18 Body weight Intra-abdominal Food consumption after
feeding fat mass/body (n = 8/group, Mean .+-. SD) (g/day/mouse) (g)
weight (%) Non-treatment group 3.17 .+-. 0.43 22.9 .+-. 1.5 1.76
.+-. 0.73 (Control group) 1% Cinnamon 2.61 .+-. 0.38 21.5 .+-. 1.0
1.00 .+-. 0.27 extract group
[0120] It is apparent from Table 18 that compared with the
non-treatment group (control group), the cinnamon extract group
showed no difference in food consumption or in body weight but the
intra-abdominal fat mass was remarkably decreased in this group. It
was clear that the visceral fat accumulated by the intake of the
high-fat, high-carbohydrate food was decreased by the intake of the
cinnamon extract-supplemented food.
34 (Example 16) Preparation of cinnamon extract-containing tablets
Cinnamon extract 45 weight parts Lactose 35 weight parts
Crystalline cellulose 15 weight parts Sucrose fatty acid ester 5
weight parts
[0121] According to the above recipe, cinnamon extract-containing
tablets for food/beverage or medical use were manufactured by the
established procedure.
INDUSTRIAL APPLICABILITY
[0122] The present invention, constituted as above, provides a
composition for preventing or ameliorating multiple risk factor
syndrome involving visceral fat-type obesity, diabetes mellitus,
hyperlipemia, and hypertension. Intake of the composition of the
invention results in reductions in accumulated visceral fat and,
hence, leads to preventing or ameliorating lifestyle-related
diseases arising from accumulation of visceral fat, such as
obesity, diabetes mellitus, hyperlipemia, and hypertension.
* * * * *