U.S. patent application number 10/277967 was filed with the patent office on 2003-08-07 for ligand for peroxisome proliferator-activated receptor.
Invention is credited to Kishida, Hideyuki, Kitahara, Mikio, Kitano, Mitsuaki, Kuroda, Minpei, Mae, Tatsumasa, Mimaki, Yoshihiro, Nakagawa, Kaku, Sashida, Yutaka, Tsukagawa, Misuzu.
Application Number | 20030147979 10/277967 |
Document ID | / |
Family ID | 19141144 |
Filed Date | 2003-08-07 |
United States Patent
Application |
20030147979 |
Kind Code |
A1 |
Mae, Tatsumasa ; et
al. |
August 7, 2003 |
Ligand for peroxisome proliferator-activated receptor
Abstract
The object of the present invention is to provide PPAR.gamma.
ligand derived from naturally occurring sources and a composition
for preventing and/or improving Insulin Resistance Syndrome,
diabetes mellitus, obesity or visceral fat obesity comprising the
PPAR.gamma. ligand as an active agent. The present invention
relates to a ligand for peroxisome proliferator-activated receptor
which comprises curcumin or its derivatives. The composition
according to the present invention, which comprises the PPAR.gamma.
ligand as an active agent is useful for preventing and/or improving
Insulin Resistance Syndrome, diabetes mellitus, obesity or visceral
fat obesity.
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) ;
Kuroda, Minpei; (Tokyo, JP) ; Mimaki, Yoshihiro;
(Tokyo, JP) ; Sashida, Yutaka; (Tokyo,
JP) |
Correspondence
Address: |
Kenyon & Kenyon
Suite 700
1500 K Street, N.W.
Washington
DC
20005
US
|
Family ID: |
19141144 |
Appl. No.: |
10/277967 |
Filed: |
October 23, 2002 |
Current U.S.
Class: |
424/756 ;
514/679 |
Current CPC
Class: |
A23L 7/109 20160801;
A23D 9/007 20130101; A61P 3/10 20180101; A23K 50/40 20160501; A61P
43/00 20180101; A61K 31/00 20130101; A23K 20/111 20160501; A61K
31/12 20130101; A61P 3/04 20180101; A23D 7/0056 20130101; A23L
27/60 20160801; A23L 33/105 20160801; A21D 2/14 20130101 |
Class at
Publication: |
424/756 ;
514/679 |
International
Class: |
A61K 035/78; A61K
031/12 |
Foreign Application Data
Date |
Code |
Application Number |
Oct 23, 2001 |
JP |
2001-324414 |
Claims
What we claim is:
1. A ligand for peroxisome proliferator-activated receptor which
comprises curcumin or its derivative.
2. The ligand for peroxisome proliferator-activated receptor
according to claim 1, wherein said peroxisome
proliferator-activated receptor is a peroxisome
proliferator-activated receptor .gamma..
3. A composition for preventing and/or improving Insulin Resistance
Syndrome, which comprises at least one selected from the group
consisting of curcumin and its derivatives as an active agent.
4. A composition for preventing and/or improving diabetes mellitus,
which comprises at least one selected from the group consisting of
curcumin and its derivatives as an active agent.
5. A composition for preventing and/or improving obesity or
visceral fat obesity, which comprises at least one selected from
the group consisting of curcumin and its derivatives as an active
agent.
6. The composition according to any of claims 3 to 5, which is
applicable for foods and drinks.
7. The composition according to any of claims 3 to 5, which is
applicable for pharmaceuticals.
8. The composition according to any of claims 3 to 5, which can be
given to domestic animals or pets.
Description
BACKGROUND OF THE INVENTION
[0001] The present invention relates to a ligand for peroxisome
proliferator-activated receptor, and a composition for preventing
and/or improving Insulin Resistance Syndrome (e.g., type-II
diabetes mellitus, hyperinsulinemia, dyslipidemia, obesity,
hypertension and arteriosclerotic cardiovascular disease)
comprising the ligand for peroxisome proliferator-activated
receptor as an active agent.
PRIOR ART
[0002] Peroxisome proliferator-activated receptor (PPAR) is a
ligand-dependent transcriptional regulatory factor belonging to the
nuclear receptor family, which has been identified as a
transcriptional regulatory factor that regulates expression of a
group of genes that maintain lipid metabolism. It is known that
three subtypes of PPAR, i.e., PPAR.alpha., PPAR.delta. (PPAR.beta.,
NUC-1, FAAR) and PPAR.gamma., have been identified in mammals.
PPAR.alpha. is mainly expressed in the liver while PPAR.delta. is
ubiquitously expressed. PPAR.gamma. has two isoforms, PPAR.gamma.1
and PPAR65 2. PPAR.gamma.1 is expressed not only in adipose tissues
but also in immune system organs, adrenals and small intestine.
PPAR.gamma.2 is specifically expressed in adipose tissues, and is a
master regulator which regulates differentiation/maturation of
adipocytes (Teruo Kawada, Igaku no Ayumi (Journal of Clinical and
Experimental Medicine) 184, 519-523, 1998).
[0003] Examples of known PPAR.gamma. ligands include: arachidonic
acid metabolites such as 15-deoxy-.DELTA.12, 14-prostaglandin J2
and .DELTA.12-prostaglandin J2; unsaturated fatty acids such as
.omega.-3-polyunsaturated fatty acid, .alpha.-linolenic acid,
eicosapentaenoic acid (EPA) and docosahexaenoic acid (DHA); and
eicosanoids such as 9-hydroxy-octadecadienoic acid and
13-hydroxy-octadecadienoic acid (J. Auwerx, Diabetologia, 42,
1033-1049, 1999). It has been also disclosed that PPAR.gamma.
ligands include C.sub.10-26 conjugated-unsaturated fatty acids
having a conjugated-triene or tetraene structure (Japanese Patent
Application 2000-355538). It is also known that examples of
synthetic PPAR.gamma. ligands include thiazolidinediones such as
troglitazone, pioglitazone and rosiglitazone.
[0004] It has been suggested that thiazolidinediones, which are
PPAR.gamma. ligands, are associated with improvement of insulin
resistance since their agonistic activities correlates with their
hypoglycemic actions. Based on these findings, they were developed
as drugs for improving insulin resistance against type-II diabetes
mellitus (non-insulin dependent diabetes mellitus: NIDDM). Namely,
a thiazolidinedione, which is one of PPAR.gamma. ligands, can
improve insulin resistance by activating PPAR.gamma. to increase
number of small adipocytes with normal function differentiated from
preadipocytes and to decrease number of large adipocytes, which
hyperproduce and/or hypersecrete factors causing insulin resistance
such as TNF.alpha. and free fatty acid, by apoptosis (A. Okuno, et
al., Journal of Clinical Investigation, 101, 1354-1361, 1998).
PPAR.gamma. ligands are also useful for prevention and/or
improvement of Insulin Resistance Syndrome, not only for type-II
diabetes mellitus but also for hyperinsulinemia, dyslipidemia,
obesity, hypertension and arteriosclerotic cardiovascular disease
(R. A. DeFronzo, et al., Diabetes Care, 14, 173-194, 1991), due to
its ability to improve insulin resistance. As for the effect
against obesity, it has been reported that administration of
troglitazone to type-II diabetic patients reduces visceral fat in
the patients (I. E. Kelly, et al., Diabetes Care, 22, 288-293,
1999; Y. Mori, et al., Diabetes Care, 22, 908-912, 1999). Thus,
PPAR.gamma. ligands are also useful for prevention and/or
improvement of visceral fat obesity.
[0005] Curcumin and its derivatives are components contained in
tropical or subtropical plants, of which a good representative is
perennial Curcuma longa, belonging to Zingiberaceae. Curcuma longa
is generally known as turmeric, one of spices which are used in
curry, and can be used not only for foods, but also as a colorant
in food or clothing, or as a herbal medicine in traditional
therapies such as Chinese medicine (Kampo), Indian Ayurveda and
Indonesian Jamu due to its hemostatic, stomachic, antibacterial and
anti-inflammatory actions.
[0006] It has been proved that curcumin has various physiological
activities such as anti-oxidative action, cholagogic action, the
internal organs (hepatic or pancreatic) function-potentiating
action, carcinogenesis-inhibiting action, lipid
metabolism-improving action, and whitening action. P. Suresh Babu
and K. Srinivasan reported that streptozotocin-induced diabetic
rats, which were maintained on diet containing 0.5% curcumin,
exhibited reduced cholesterol, triglyceride and phospholipid levels
in blood (Molecular and Cellular Biochemistry, 166, 169-175, 1997)
and amelioration of renal lesions associated with diabetes mellitus
(Molecular and Cellular Biochemistry, 181, 87-96, 1998). Japanese
Patent Application Hei-11-246399 discloses that enhanced activity
of acyl-CoA oxidase (.beta.-oxidation promotive enzyme) and
inhibition of triglyceride accumulation in the liver were observed
in rats which received curcumin. However, it has not been known
that curcumin and/or its derivatives are PPAR.gamma. ligands and
have hypoglycemic or visceral fat-reducing action.
[0007] As described above, PPAR.gamma. ligands can improve insulin
resistance and prevent and/or improve Insulin Resistance Syndrome
such as type-II diabetes mellitus, hyperinsulinemia, dyslipidemia,
obesity (particularly visceral fat obesity) , hypertension and
arteriosclerotic cardiovascular disease. Accordingly, the object of
the present invention is to provide a PPAR.gamma. ligand derived
from naturally occurring sources and a composition comprising the
PPAR.gamma. ligand as an active agent for preventing and/or
improving Insulin Resistance Syndrome, diabetes mellitus, obesity
or visceral fat obesity.
SUMMARY OF THE INVENTION
[0008] The present inventors found that Curcuma extract has
hypoglycemic action and that, after intense studies, particular
components contained in Curcuma (curcumin and its derivatives) have
PPAR.gamma. ligand activities. They also found that these
particular components have hypoglycemic action and visceral
fat-reducing action. The present invention was developed based on
these findings.
[0009] In summary, the present invention relates to a ligand for
peroxisome proliferator-activated receptor
[0010] which comprises curcumin or its derivative.
[0011] The present invention also relates to a composition for
preventing and/or improving Insulin Resistance Syndrome, diabetes
mellitus, or obesity or visceral fat obesity
[0012] which comprises at least one selected from the group
consisting of curcumin and its derivatives as an active agent.
DESCRIPTION OF THE PREFERRED EMBODIMENTS
[0013] The present invention will be described in more detail
referring to the following embodiments. The PPAR.gamma. ligand
according to the present invention comprises curcumin or its
derivative. A composition comprising, as an active agent, at least
one selected from the group consisting of curcumin and its
derivatives according to the present invention has hypoglycemic
action and visceral fat-reducing action, and therefore be useful to
prevent and/or improve Insulin Resistance Syndrome, diabetes
mellitus, obesity or visceral fat obesity.
[0014] Curcumin to be used in the present invention is 1,7,-bis
(4-hydroxy-3-methoxyphenyl)-1,6-heptadiene-3,5-dione, and curcumin
derivatives (curcuminoids) include, for example, demethoxycurcumin,
bisdemethoxycurcumin, dihydrocurcumin, tetrahydrocurcumin,
hexahydrocurcumin, dihydroxytetrahydrocurcumin, Yakuchinone A and
Yakuchinone B, and their salts, oxidants, reductants, glycosides
and esters thereof. Those may be purified from plants or chemically
synthesized compounds. Plant-derived curcumin and/or its
derivatives can be obtained by extraction from plants including
Zingiberaceae Curcuma, such as Curcuma longa(turmeric), Curcuma
aromatica(wild turmeric), Curcuma zedoaria (zedoary), Curcuma
xanthorrhiza, mango ginger, Indonesian arrowroot, yellow zedoary,
black zedoary and galangal.
[0015] Any conventional method can be used to prepare curcumin and
its derivatives to be used in the present invention. For example,
turmericoleoresin, a food additive, which essentially contains
curcumin, can be produced by extracting from a dry product of
rhizome of turmeric with ethanol at an elevated temperature, with
hot oil and fat or propylene glycol, or with hexane or acetone at
from room temperature to a high temperature. Alternatively, those
can be produced by the methods disclosed in Japanese Patent
Application 2000-236843, Japanese Patent Application Hei-11-235192
and Japanese Patent Application Hei-6-9479, and Japanese Kohyo
Publication Hei-11-502232 and Japanese Kohyo Publication
Hei-9-503528. According to the present invention, a purified
product of at least one selected from the group consisting of
curcumin and its derivatives may be used. Alternatively, a
semi-purified or crude product thereof may be used, provided that
it does not contain impurities which may not be acceptable as a
pharmaceutical or food product.
[0016] A composition for preventing and/or improving Insulin
Resistance Syndrome, diabetes mellitus, obesity or visceral fat
obesity according to the present invention may comprise a
PPAR.gamma. ligand, and is the composition comprises, as an active
agent, at least one selected from the group consisting of curcumin
and its derivatives. The compositions of the invention may be used
in, for example, but not limited to, foods and drinks including
foods with health claims (e.g., foods for specified health uses or
foods for nutrient function claims) or health foods,
pharmaceuticals and quasi drugs.
[0017] When used as foods and drinks, the inventive compositions
may be administered alone, or formulated in combination with any
known carrier(s) and/or additive(s) into any suitable dosage form
including, for example, capsules, tablets and granules. The
PPAR.gamma. ligand according to the present invention may be
present in such formulations at an amount of 0.1 to 100% by weight,
and preferably 10 to 90% by weight. Alternatively, the inventive
composition may be added to any kinds of foods and drinks,
including: confectionery such as chewing gums, chocolates, candies,
jellies, biscuits or crackers; frozen desserts such as an ice cream
or ice cube; drinks such as tea, soft drinks, nutritional
supplement drinks or beauty supplement drinks; noodles such as udon
noodle, Chinese noodle, spaghetti or instant noodle; foods made
from fish paste such as boiled fish paste (kamaboko), tube-shaped
fish paste cake (chikuwa) or a cake of pounded fish (hanpen);
dressing, mayonnaise, sauce or other seasonings; fat foods such as
margarine, butter or salad oil; bread; ham; soup; boil-in-bag
foods; and frozen foods. A food or drink containing the inventive
composition may be given to a human at a dose of 0.1 to 3000 mg/kg
body weight/day (based on inventive PPAR.gamma. ligand) for an
adult, and preferably 1 to 300 mg/kg body weight/day (based on the
PPAR.gamma. ligand). The inventive composition may be also used as
a feed for a domestic animal or pet-food for a pet. In this case, a
feed or food containing the inventive composition may be preferably
given at a dose of 0.1 to 3000 mg/kg body weight/day (based on the
PPAR.gamma. ligand).
[0018] When used as pharmaceuticals, the inventive compositions may
be formulated into any suitable dosage forms for administration
including, but not limited to, capsules, tablets, granules,
injection solution, suppositories and patches. In preparation of
the drugs, such formulations comprising the inventive composition
may additionally comprise other pharmaceutically acceptable
additive(s) such as an excipient, a disintegrator, a lublicant, a
binder, an anti-oxidant, a colorant, an anti-aggregation agent, a
sorbefacient, a solubilizer and/or a stabilizer as appropriate.
Such a formulation may be administered to a human at a dose of 0.1
to 3000 mg/kg body weight/day (based on inventive PPAR.gamma.
ligand) for an adult, and preferably 1 to 300 mg/kg body weight/day
(based on PPAR.gamma. ligand), once or divided into several times a
day. The inventive composition may be also administered to a
domestic animal or a pet as a pharmaceutical drug. In this case, a
formulation containing the inventive composition may be preferably
administered at a dose of 0.1 to 3000 mg/kg body weight/day (based
on the inventive PPAR.gamma. ligand).
[0019] According to the present invention, a ligand for peroxisome
proliferator-activated receptor .gamma. (PPAR.gamma. ) and a
composition comprising the same are provided. The composition
according to the present invention is useful for preventing and/or
improving Insulin Resistance Syndrome, diabetes mellitus, obesity
or visceral fat obesity.
EXAMPLES
[0020] The present invention will be described in more detail by
referring to the following Examples though the present invention is
not limited to these Examples.
Example 1
Extraction and Isolation of Compounds from Curcuma longa
[0021] Curcuma longa powder (1.0 kg) was extracted with ethanol
(8.0 L) at room temperature in darkness for 2 days and filtered to
give an extracted solution. The solvent was removed from the
extracted solution by vacuum concentration to give an extract (118
g). The extract was then subjected to porous ion-exchange resin
DIAION HP-20 column chromatography (1600 ml), and eluted
sequentially with 30% methanol, 50% methanol and 80% methanol (1.5
L each), and then methanol, ethanol and ethyl acetate (3 L each) to
give 6 fractions (fractions 1, 2, 3, 4, 5 and 6). Purification was
performed by repeatedly subjecting fraction 4 (63.5 g) to silica
gel chromatography A (eluant; hexane:
acetone=2:1.fwdarw.3:2.fwdarw.4:3, v/v) and silica gel
chromatography B (eluant; chloroform: acetone=99:1.fwdarw.19:1,
v/v) to give compound 1 (6.4 g) compound 2 (1.2 g) and compound 3
(1.1 g).
[0022] Structure analysis showed that compounds 1 to 3 were known
compounds: compound 1 was identified as curcumin, compound 2 was
demethoxycurcumin and compound 3 was bisdemethoxycurcumin,
respectively. The structures of these compounds were identified
based on the spectrum data described in the report by M. Kuroyanagi
et al. (Yakugaku Zassi (Journal of Pharmaceuticals Society of
Japan), 90,1467-1470, 1970). Structural formulae of compounds 1 to
3 are shown in Table 1 below.
1TABLE 1 Compound No. Compound Name Structural Formula Compound 1
Curcumin 1 Compound 2 Demethoxycurcumin 2 Compound 3
Bisdemethoxycurcumin 3
Example 2
PPAR.gamma. Ligand Activity
[0023] CV-1 cells (culture cells derived from male African green
monkey kidney) were inoculated into a 96-well culture plate at
6.times.10.sup.3 cells/well, and incubated at 37.degree. C. for 24
hours under 5% CO.sub.2 condition. As a medium, a DMEM (Dulbecco's
Modified Eagle Medium, product of GIBCO) containing 10% FBS (fetal
bovine serum), 10 ml/L penicillin-streptomycin (5000 IU/ml and 5000
.mu.g/ml, respectively, product of GIBCO), 37 mg/L ascorbic acid
(product of Wako Pure Chemical Industries, Ltd.) was used. Cells
were washed with OPTI-MEM (product of GIBCO), and transfected with
pM-mPPAR.gamma. and 4.times.UASg-luc using LipofectAMINE PLUS.TM.
(product of GIBCO). The pM-mPPAR.gamma. is a plasmid for chimeric
protein expression which consisted of a yeast-derived transcription
factor GAL4 gene (amino acid sequences 1 to 147) ligated to a mouse
PPAR.gamma. ligand binding domain gene (amino acid sequences 174 to
475). The 4.times.UASg-luc is a reporter plasmid incorporated
luciferase gene with 4-time-repeated responsive element (UASg) of
GAL4 ligated thereto at the upstream end thereof. At approximately
24 hours after the transfection, the medium was replaced by a
medium containing each sample (n=4) and cells were incubated for
additional 24 hours. Each sample used was dissolved in dimethyl
sulfoxide (DMSO) and DMSO was used as an untreated control sample.
These samples were added to the medium at a volume ratio of 1/1000.
Cells were washed with Ca, Mg-containing phosphate buffered saline
(PBS+) and added with LucLite.TM. (product of Packard). Then, the
luminescent intensity by expressed luciferase was determined in a
TopCount.TM. Microplate Scintillation/Luminescence Counter (product
of Packard).
[0024] The luminescent intensity was determined in the control
group in the same way as in the test groups, but using pM (a
plasmid lacking PPAR.gamma. ligand binding domain gene) instead of
pM-mPPAR.gamma.. The ratio (test group/control group) of the
average luminescent intensity between the test and control groups
(n=4) was determined for each sample, and the relative activity of
test sample against the untreated control sample was determined as
the PPAR.gamma. ligand activity of the sample. The results are
shown in Table 2 below.
2 TABLE 2 PPAR.sub..gamma. Added Concentration ligand activity
Untreated Control (DMSO) (0.1%) 1.00 Troglitazone 0.5 .mu.M 2.37
.+-. 0.24 1 .mu.M 4.26 .+-. 0.22 2 .mu.M 6.93 .+-. 0.56 Compound 1
2 .mu.g/ml (5.4 .mu.M) 2.36 .+-. 0.94 (Curcumin) 5 .mu.g/ml (13.6
.mu.M) 3.67 .+-. 1.06 10 .mu.g/ml (27.2 .mu.M) 4.16 .+-. 1.10
Compound 2 2 .mu.g/ml (5.9 .mu.M) 2.17 .+-. 0.39
(Demethoxycurcumin) 5 .mu.g/ml (14.8 .mu.M) 4.25 .+-. 0.77 10
.mu.g/ml (29.6 .mu.M) 4.41 .+-. 0.35 Compound 3 2 .mu.g/ml (6.5
.mu.M) 1.88 .+-. 0.33 (Bisdemethoxycurcumin) 5 .mu.g/ml (16.2
.mu.M) 3.55 .+-. 0.38 10 .mu.g/ml (32.5 .mu.M) 4.08 .+-. 0.46 (Mean
.+-. SD)
[0025] PPAR.gamma. ligand activity of each test compound was
compared with that of troglitazone (product of Sankyo) used as a
positive control. It is apparent from Table 2 that curcumin,
demethoxycurcumin and bisdemethoxycurcumin exhibited
concentration-dependent PPAR.gamma. ligand activities.
Example 3
The Effect of the Compound in Model Mice of Type-II Diabetes
Mellitus
[0026] KK-Ay mice, a model of genetically obese and type-II
diabetic animals, were used to evaluate the effect of curcumin.
Pioglitazone, a drug for treating diabetes mellitus, was used as a
positive control.
[0027] KK-Ay mice (females, 6 weeks old) were divided into 3 groups
(5 animals per group). By using a normal diet (product of Oriental
Yeast Co. Ltd., Table 3) as a base feed, three types of feeds,
i.e., a diet without any additives, with pioglitazone and with
curcumin, were prepared. Mice were placed in an environment in
which they were freely accessible to a diet without any additives
(control group), with pioglitazone (pioglitazone-added group) or
with curcumin (curcumin-added group) for 4 weeks. Pioglitazone used
was obtained by grinding Actos tablet 30 (30 mg pioglitazone per
tablet, Takeda Chemical Industries Ltd.) in an agate mortar. The
ground Actos tablet was then added to the normal diet at a dose of
0.04% pioglitazone. Curcumin was added to the normal diet at a dose
of 0.5% curcumin.
3 TABLE 3 Normal diet (AIN-93G modified) Ratio Fat 22% Carbohydrate
58.5% Protein 19.5% Total energy 4,100 kcal/kg Formulation Casein
20.000% Cornstarch 49.948% Sucrose 10.000% Soybean oil 10.000%
Cellulose powder 5.000% AIN-93 mineral mixture 3.500% AINl93
vitamin mixture 1.000% Choline bitartrate 0.250% Tertiary
butylhydroquinone 0.002% L-cystine 0.300%
[0028] During feeding, a small amount of blood was collected from
the caudal vein of mice every week to determine blood glucose level
using a simple blood glucose level analyzer GLUTESTACE (Sanwa
Kagaku Kenkyusho Co., Ltd.).
[0029] The mouse body weights are shown in Table 4. The mouse body
weights in both pioglitazone-added and curcumin-added groups
changed in a similar pattern of control group (without any
additives) without any significant difference.
4 TABLE 4 Mouse Body weight (g) Control group (without any
Pioglitazone-added Curcumin-added additives) group group Start 27.3
.+-. 0.3 26.9 .+-. 1.0 25.5 .+-. 0.7 After 1 week 34.5 .+-. 0.9
36.7 .+-. 1.0 34.7 .+-. 1.1 After 2 weeks 38.9 .+-. 1.1 39.9 .+-.
1.2 39.8 .+-. 0.7 After 3 weeks 41.0 .+-. 1.3 42.3 .+-. 1.2 42.8
.+-. 0.9 After 4 weeks 43.5 .+-. 1.3 43.8 .+-. 1.4 44.7 .+-. 1.0
(Mean .+-. SD)
[0030] Blood glucose levels are shown in Table 5 below. When
feeding started, mice had a blood glucose level of 139 to 151
mg/dl, and hyperglycemia was not observed in any groups. Mice of
the control group (without any additives) showed elevated blood
glucose level, indicating development of diabetes mellitus.
Elevation of blood glucose level observed in the mice of the group
which received pioglitazone (a drug for treating diabetes mellitus)
was suppressed significantly, compared with that of the control
group (without any additives), indicating potent hypoglycemic
action of pioglitazone. In the curcumin group, the elevation of
blood glucose level was also suppressed significantly, indicating
that curcumin has hypoglycemic action.
5 TABLE 5 Blood Glucose Level (mg/dl) Control group (without any
Pioglitazone-added Curcumin-added additives) group group Start 142
.+-. 12 151 .+-. 9 139 .+-. 7 After 1 weeks 322 .+-. 70 163 .+-. 23
** 191 .+-. 18 ** After 2 weeks 427 .+-. 70 182 .+-. 9 ** 222 .+-.
46 ** After 3 weeks 455 .+-. 66 166 .+-. 18 ** 348 .+-. 125 After 4
weeks 479 .+-. 71 153 .+-. 21 ** 344 .+-. 105 * (Mean .+-. SD: *p
< 0.05; **p < 0.01)
Example 4
The Effect of the Compound in Model Mice of Diet-induced
Obesity
[0031] C57BL/6J mice (females, 8 weeks old) were freely accessible
to a high fat/high sugar diet (Oriental Yeast Co., Ltd., Table 6)
for 8 weeks to obtain dietary obese animals. By using a normal diet
(Oriental Yeast Co. Ltd., Table 3) as a base feed, two types of
feeds, i.e., a diet without any additives and with 0.5% curcumin,
were prepared. Next, said mice were divided into 2 groups (7
animals per group), and each group was freely accessible to a diet
without any additives (control group) or with 0.5% curcumin
(curcumin group) for 4 weeks. After an overnight fasting, mice were
subjected to abdominal section under ether anesthesia to collect
blood from the abdominal aorta, and then sacrificed. Then, adipose
tissues were collected from the tissues around uterus, kidney and
mesentery, and their weights were determined. The sum of the
weights of periuterine, perirenal and mesenteric adipose tissues
was determined as the total amount of intra-abdominal adipose
tissue. The results are shown in Table 7.
6 TABLE 6 High fat/High sugar divisional diet Ratio Fat 53%
Carbohydrate 27% Protein 20% Total energy 5,100 kcal/kg Formulation
Casein 25.000% Cornstarch 14.869% Sucrose 20.000% Soybean oil
2.000% Lard 14.000% Beef tallow 14.000% Cellulose powder 5.000%
AIN-93 mineral mixture 3.500% AINl93 vitamin mixture 1.000% Choline
bitartrate 0.250% Tertiary butylhydroquinone 0.006% L-cystine
0.375%
[0032]
7 TABLE 7 Control group (without any Curcumin-added additives)
group Diet intake amount (g/day/animal) 3.16 .+-. 0.58 3.24 .+-.
0.55 Body weight after feeding (g) 24.4 .+-. 2.5 22.9 .+-. 0.9
Adipose tissue per body weight (% by weight) Periuterine adipose
tissue (a) 1.64 .+-. 0.82 0.8 .+-. 0.21 ** Perirenal adipose tissue
(b) 0.86 .+-. 0.50 0.46 .+-. 0.15 * Mesenteric adipose tissue (c)
0.75 .+-. 0.43 0.39 .+-. 0.11 * Intra-abdominal adipose tissue 3.25
.+-. 1.73 1.65 .+-. 0.42 ** (a + b + c) (Mean .+-. SD; *p <
0.05; **p < 0.01)
[0033] It is apparent from Table 7 that no significant difference
was detected in diet intake amount or body weight between the
curcumin group and the control group (without any additives) while
the curcumin group exhibited significantly reduced the weights of
periuterine, perirenal, mesenteric and intra-abdominal adipose
tissues when compared with those of the control group. In other
words, it was proved that intake of curcumin-containing food
reduces the visceral fat accumulated by taking high-fat/high sugar
diet.
8EXAMPLE 5 Preparation of curcumin-containing tablets Curcumin 45
parts by weight Lactose 35 parts by weight Crystalline cellulose 15
parts by weight Sucrose fatty acid ester 5 parts by weight
[0034] Curcumin-containing tablets for foods were prepared using
the above-listed ingredients according to a conventional
method.
9EXAMPLE 6 Preparation of curcumin-containing soft capsules
Curcumin 40 parts by weight Sesame oil 55 parts by weight Glycerin
fatty acid ester 5 parts by weight
[0035] Curcumin-containing soft capsules for foods were prepared
using the above-listed ingredients according to a conventional
method.
10EXAMPLE 7 Preparation of curcumin-containing crackers Curcumin 1
part by weight Plain flour 120 parts by weight Salt 1 part by
weight Baking powder 2 parts by weight Butter 30 parts by weight
Water 40 parts by weight
[0036] Curcumin-containing crackers were prepared using the
above-listed ingredients according to a conventional method.
11EXAMPLE 8 Preparation of curcumin-containing udon noodle Curcumin
1 part by weight Bread flour 100 parts by weight Plain flour 100
parts by weight Salt 10 parts by weight Water 100 parts by
weight
[0037] Curcumin-containing udon noodle was prepared using the
above-listed ingredients according to a conventional method.
12EXAMPLE 9 Preparation of curcumin-containing dressing Curcumin 10
parts by weight Olive oil 80 parts by weight Vinegar 60 parts by
weight Salt 3 parts by weight Pepper 1 part by weight Lemon juice 5
parts by weight
[0038] Curcumin-containing dressing was prepared using the
above-listed ingredients according to a conventional method.
* * * * *