U.S. patent application number 14/630911 was filed with the patent office on 2015-06-18 for cholecystokinin secretion-promoting composition.
The applicant listed for this patent is J-OIL MILLS, INC.. Invention is credited to Hiroshi HARA, Tohru HIRA, Jun IMAGI, Chigusa NISHIYAMA, Noriko TOKURA, Takatoshi YAMASHITA.
Application Number | 20150164823 14/630911 |
Document ID | / |
Family ID | 50278016 |
Filed Date | 2015-06-18 |
United States Patent
Application |
20150164823 |
Kind Code |
A1 |
HARA; Hiroshi ; et
al. |
June 18, 2015 |
CHOLECYSTOKININ SECRETION-PROMOTING COMPOSITION
Abstract
A CCK secretion-promoting substance and a use thereof are
provided. This cholecystokinin secretion-promoting composition
contains, as an active component, acrylic acid and/or an
unsaturated aldehyde having a main chain of 4-12 carbon atoms
having a double bond in at least position 2 or 4, wherein the main
chain has 4-9 carbon atoms if there is a double bond only in
position 2 and the main chain has 9-12 carbon atoms if there is a
double bond only in position 4. This cholecystokinin
secretion-promoting composition can be used as an appetite
suppressant. By adding this cholecystokinin secretion-promoting
composition to food, appetite-suppressing food products can be
provided.
Inventors: |
HARA; Hiroshi; (Hokkaido,
JP) ; HIRA; Tohru; (Hokkaido, JP) ; NISHIYAMA;
Chigusa; (Tokyo, JP) ; TOKURA; Noriko; (Tokyo,
JP) ; YAMASHITA; Takatoshi; (Tokyo, JP) ;
IMAGI; Jun; (Tokyo, JP) |
|
Applicant: |
Name |
City |
State |
Country |
Type |
J-OIL MILLS, INC. |
Tokyo |
|
JP |
|
|
Family ID: |
50278016 |
Appl. No.: |
14/630911 |
Filed: |
February 25, 2015 |
Related U.S. Patent Documents
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Application
Number |
Filing Date |
Patent Number |
|
|
PCT/JP2013/068827 |
Jul 10, 2013 |
|
|
|
14630911 |
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Current U.S.
Class: |
514/703 ;
562/598; 568/448 |
Current CPC
Class: |
A61P 43/00 20180101;
A61K 31/19 20130101; A23L 33/10 20160801; A23V 2002/00 20130101;
A61K 31/11 20130101; A61K 9/0056 20130101; A61P 3/04 20180101; C07C
47/21 20130101 |
International
Class: |
A61K 31/11 20060101
A61K031/11; A61K 31/19 20060101 A61K031/19 |
Foreign Application Data
Date |
Code |
Application Number |
Sep 14, 2012 |
JP |
2012202450 |
Claims
1. A cholecystokinin secretion-promoting composition comprising, as
an active component, acrylic acid and/or an unsaturated aldehyde
having a main chain of 4 to 12 carbon atoms having a double bond in
at least position 2 or 4, wherein the main chain has 4 to 9 carbon
atoms if there is a double bond in only position 2, and the main
chain has 9 to 12 carbon atoms if there is a double bond in only
position 4.
2. The cholecystokinin secretion-promoting composition according to
claim 1, wherein the unsaturated aldehyde is selected from the
group consisting of di-unsaturated aldehydes having 4 to 12
main-chain carbon atoms and having double bonds in positions 2 and
4, mono-unsaturated aldehydes having 4 to 9 main-chain carbon atoms
and having a double bond in position 2, and mono-unsaturated
aldehydes having 9 to 12 main-chain carbon atoms and having a
double bond in position 4.
3. The cholecystokinin secretion-promoting composition according to
claim 1, comprising the unsaturated aldehyde having a double bond
in position 2.
4. The cholecystokinin secretion-promoting composition according to
claim 1, comprising the unsaturated aldehyde having double bonds in
positions 2 and 4.
5. The cholecystokinin secretion-promoting composition according to
claim 1, wherein the unsaturated aldehyde is in a trans
conformation.
6. The cholecystokinin secretion-promoting composition according to
claim 1, comprising the unsaturated aldehyde having two double
bonds and the unsaturated aldehyde having one double bond, or
comprising the unsaturated aldehyde having two different double
bonds.
7. The cholecystokinin secretion-promoting composition according to
claim 6, wherein the unsaturated aldehyde having one double bond is
trans-2-octenal.
8. An appetite suppressant comprising the cholecystokinin
secretion-promoting composition according to claim 1.
9. An appetite-suppressing food product comprising the
cholecystokinin secretion-promoting composition according to claim
1.
Description
TECHNICAL FIELD
[0001] The present invention relates to a cholecystokinin
secretion-promoting composition, more specifically, a
cholecystokinin secretion-promoting composition useful as an
appetite suppressant.
BACKGROUND
[0002] The term "obesity" refers to a state of having a larger body
weight or a state of having an excessive accumulation of body fat
compared to a normal state. Obesity is a contemporary lifestyle
disease caused by biased dietary habits, lack of exercise, or lack
of sleep. Obesity caused by a metabolic disorder or an endocrine
disease is called symptomatic obesity. Either obesity can be a risk
factor of diseases such as hyperlipidemia, hypercholesterolemia,
hypertriglyceridemia, hypertension, arteriosclerosis, ischemic
heart disease, stroke, and arteriosclerosis obliterans.
Accordingly, prophylaxis or treatment of obesity can be an
effective means for preventing these diseases.
[0003] Cholecystokinin (hereinafter, referred to as CCK) is a
gastrointestinal hormone secreted from the duodenal mucosal cells
by ingestion of a lipid, protein, or other nutrients. This hormone
is known to accelerate the enzyme secretion from the pancreas, to
contract the gallbladder, and to delay the transfer of gastric
contents to the duodenum by closing the pylorus of the stomach
(Non-Patent Literature 1). This hormone also directly acts on the
central nervous system to suppress the appetite. Thus, the hormone
has a physiological function of inducing a feeling of fullness.
[0004] By focusing on the physiological effects of CCK, materials
having a CCK secretion stimulating activity have been searched, and
the uses of the materials have been developed. For example, Patent
Literature 1 (Arginine-containing peptide having a cholecystokinin
secretion-stimulating activity and food containing the peptide)
discloses that a pepsin digestion product of soybean
.beta.-conglycinin promotes the CCK-secreting activity in rats to
reduce the food intake and that the soybean .beta.-conglycinin is
used in a food intake-suppressing food. Non-Patent Literatures 2
and 3 ("Soybean .beta.-Conglycinin Bromelain Hydrolysate Stimulates
Cholecystokinin Secretion by Enteroendocrine STC-1 Cells to
Suppress the Appetite of Rats under Meal-Feeding Conditions" and
"Acute effect of soybean beta-conglycinin hydrolysate ingestion on
appetite sensations in healthy humans") report on that a bromelain
hydrolysate of soybean .beta.-conglycinin having a CCK
secretion-promoting action suppresses the appetite of rats and
human beings. Non-Patent Literature 4 (Development of peptide
regulating appetite by controlling a gastrointestinal hormone with
high safety) reports on a food containing a bromelain digestion
product or peptide of .beta.-conglycinin.
[0005] Patent Literature 2 (Composition containing pork-derived
peptide having a food intake-suppressing action) discloses a
composition containing a peptide having a cholecystokinin
secretion-stimulating activity or food intake-suppressing activity
prepared by digesting pork or pork-derived protein with pepsin on
the basis that the pig-derived peptide promotes CCK secretion and
reduces the food intake of rats.
[0006] Patent Literature 3 (Pharmacological composition for
suppressing appetite) discloses a pharmacological composition for
suppressing appetite containing an ingredient showing a
cholecystokinin secretion-stimulating action prepared from yeast.
This ingredient is low in calorie and has high heat resistance and
high enzymolysis resistance.
[0007] The list of patent literature and non-patent literature
documents as described herein is as follows:
PATENT LITERATURE
[0008] Patent Literature 1: Japanese Patent Laid-Open No.
2004-10569 [0009] Patent Literature 2: Japanese Patent Laid-Open
No. 2007-230978 [0010] Patent Literature 3: Japanese Patent
Laid-Open No. 2009-84191
NON-PATENT LITERATURE
[0010] [0011] Non-Patent Literature 1: Eiyo Kino Kagaku
(Nutritional Biochemistry), p. 42, (edited by Eiyo Kino Kagaku
Kenkyukai, Asakura Publishing Co., Ltd., published in 1996) [0012]
Non-Patent Literature 2: Biosci. Biotechnol. Biochem., 75(5),
848-853, 2011 [0013] Non-Patent Literature 3: Appetite, 57(3),
765-768, 2011 [0014] Non-Patent Literature 4:
http://www.naro.affrc.go.jp/brain/ibunya/files/2006.sub.--7
syokuyoku.pdf
SUMMARY
[0015] It has been demonstrated that there are CCK
secretion-promoting materials, such as a degradation product of
.beta.-conglycinin and a pig-derived peptide, and that these
materials cause advantageous effects, such as a delay in the
excretion of gastric contents, a reduction in food intake, and an
increase in the feeling of fullness, in animals including human
beings.
[0016] Accordingly, it is an object of the present invention to
search a new orally ingestible CCK secretion-promoting material and
to provide a use of the material.
[0017] During studies on the mechanism of promoting the secretion
of CCK in cultured cells by food ingredients, the present inventors
have surprisingly found that acrylic acid and specific unsaturated
aldehydes have a CCK secretion-stimulating activity and have
accomplished the present invention. That is, the present invention
provides a cholecystokinin secretion-promoting composition
(hereinafter, referred to as CCK secretion-promoting agent) as an
active component, acrylic acid and/or an unsaturated aldehyde
having a main chain of 4 to 12 carbon atoms having a double bond in
at least position 2 or 4, wherein the main chain has 4 to 9 carbon
atoms if there is a double bond in only position 2, and the main
chain has 9 to 12 carbon atoms if there is a double bond in only
position 4. Throughout the specification, the location of a double
bond is numbered in accordance with the IUPAC nomenclature.
[0018] The secretion of cholecystokinin produced by I cells in the
small intestine is physiologically promoted by peptides, amino
acids, and fatty acids in the duodenum. For example, peptides (such
as degradation products of (.beta.-conglycinin), proteins (whey and
casein), fatty acids, and calcium are conventionally known to
promote the secretion of CCK. However, it is quite a surprise that
the unsaturated aldehydes and other materials specified by the
present invention have a CCK secretion-stimulating activity.
[0019] The unsaturated aldehyde is preferably selected from the
group consisting of di-unsaturated aldehydes having 4 to 12
main-chain carbon atoms and having double bonds in positions 2 and
4, mono-unsaturated aldehydes having 4 to 9 main-chain carbon atoms
and having a double bond in position 2, and mono-unsaturated
aldehydes having 9 to 12 main-chain carbon atoms and having a
double bond in position 4.
[0020] The CCK secretion-promoting agent preferably comprises the
unsaturated aldehyde having a double bond in position 2.
[0021] The CCK secretion-promoting agent preferably comprises the
unsaturated aldehyde having double bonds in positions 2 and 4. The
unsaturated aldehydes are preferably in trans conformations.
[0022] The CCK secretion-promoting agent preferably comprises the
unsaturated aldehyde having two double bonds and the unsaturated
aldehyde having one double bond, or comprises the unsaturated
aldehyde having two different double bonds. In particular, the
unsaturated aldehyde having one double bond is preferably
trans-2-octenal.
[0023] The present invention also provides an appetite suppressant
including the CCK secretion-promoting agent.
[0024] The present invention also provides an appetite-suppressing
food product including the CCK secretion-promoting agent.
DETAILED DESCRIPTION
[0025] The present invention relates to a novel CCK
secretion-promoting agent containing a specific unsaturated
aldehyde as an active ingredient. Some examples of the unsaturated
aldehyde used in the present invention have been confirmed to be
safe as food additives. These unsaturated aldehydes are
inexpensively available and can be easily processed into a solution
or a solid. Thus, the unsaturated aldehydes are superior to
conventional CCK secretion-promoting materials in these points.
[0026] Oral ingestion of the CCK secretion-promoting agent of the
present invention promotes the activity of secreting
cholecystokinin, resulting in a reduction in food intake and also a
decrease in the sensation of hunger. Accordingly, the CCK
secretion-promoting agent of the present invention can be used as,
for example, an appetite suppressant, hyperphagia preventive agent,
or obesity preventive agent.
[0027] The present invention can also provide an
appetite-suppressing food product that induces a feeling of
fullness with a small amount thereof. The appetite-suppressing food
product can be prepared by adding a cholecystokinin
secretion-promoting composition of the present invention to a
foodstuff.
[0028] The CCK secretion-promoting agent of the present invention
comprises acrylic acid and/or an unsaturated aldehyde having 4 to
12 main-chain carbon atoms and a double bond in at least position 2
or 4. It has been revealed that acrylic acid and/or a specific
unsaturated aldehyde used in the present invention has a high
CCK-secreting activity, whereas saturated aldehydes, saturated or
unsaturated alcohols, saturated fatty acids, unsaturated fatty acid
excluding acrylic acid, and saturated or unsaturated hydrocarbons
have a low CCK secretion-stimulating activity.
[0029] The mono-unsaturated aldehyde having a double bond in only
position 2 has 4 to 9 main-chain carbon atoms. If the number of
carbon atoms in the main chain is at least 10, the CCK-secreting
activity decreases. The mono-unsaturated aldehyde having a double
bond in only position 4 has 9 to 12 main-chain carbon atoms. If the
number of carbon atoms in the main chain is not more than 8, the
CCK-secreting activity decreases.
[0030] Although an unsaturated aldehyde having three main-chain
carbon atoms and having a double bond in position 2, propenal, also
has a CCK-secreting activity, Globally Harmonized System of
Classification and Labeling of Chemicals (GHS) classifies propenal
in class 2 of acute toxicity (the swallowing of it endangers the
life). Thus, propenal is believed to be highly toxic and is
inadequate as a CCK secretion-promoting agent that is ingested to
human beings. Some unsaturated aldehydes having more than 12
main-chain carbon atoms are solid at ordinary temperature and are
predicted to cause separation or precipitation when they are used
in liquid formulations or foods. Thus, such unsaturated aldehydes
may be restricted in the use. In also unsaturated aldehydes having
a double bond in a position other than positions 2 and 4, the
CCK-secreting activity is low.
[0031] Examples of the CCK secretion-promoting agent include
trans,trans-2,4-hexadienal, trans,trans-2,4-heptadienal,
trans,trans-2,4-nonadienal, trans,trans-2,4-decadienal,
trans,trans-2,4-dodecadienal, 2,3-butadienal, 2,4-pentadienal,
3,4-pentadienal, 2,7-octadienal, 2,6-octadienal, 2,4-octadienal,
2,6-nonadienal, 4,7-decadienal, 2,4-undecadienal, 2,6-dodecadienal,
3,7-dimethyl-2,6-nonadienal, trans-2-methyl-2,6-heptadienal,
2,4-dimethyl-2,6-heptadienal, 3,6-dimethyl-2,5-heptadienal,
3,7-dimethyl-2,6-octadienal, 3,7-dimethyl-2,7-octadienal,
2,4-diethyl-2,6-heptadienal, 3,4,8-trimethyl-2,7-nonadienal,
5,9-dimethyl-4,9-decadienal, 4,8-dimethyl-4,9-decadienal,
5,9-dimethyl-4,8-decadienal,
trans-2-methyl-6-methylene-2,7-octadienal, 4,5-hexadienal,
4,7-undecadienal, trans-2-butenal, trans-2-heptenal,
trans-2-octenal, trans-2-nonenal, trans-2-pentenal,
3-methyl-2-butenal, trans-2-methyl-2-butenal, 2-ethyl-2-butenal,
4-methyl-2-pentenal, trans-2-methyl-2-pentenal, 2-methyl-2-octenal,
2-propyl-2-heptenal, 3-propyl-2-heptenal,
4-(2,5-dimethylcyclohexylidene)-2-butenal, 2-butyl-2-octenal,
5-(methylthio)-2-[(methylthio)methyl]-2-pentenal,
4-methyl-2-[(methylthio)methyl]-2-hexenal,
5-methyl-2-[(methylthio)methyl]-2-hexenal,
4-methyl-2-[(methylthio)methyl]-2-pentenal,
2-[(methylthio)methyl]-2-butenal, 2-ethyl-2-hexenal,
2-butyl-2-octenal, trans-4-decenal, cis-4-decenal, 4-nonenal,
4-dodecenal, 4-undecenal, 2-methyl-4-undecenal,
2-methyl-4-dodecenal, 2,4,6-octatrienal,
5,9-dimethyl-2,4,8-decatrienal, 2,4,6-nonatrienal,
2,4,8-undecatrienal, 2,4,6,8-undecatetraenal,
2,4,6,8-nonatetraenal, 2,4,6,8-decatetraenal,
2,4,6,8,10-undecapentaenal, and 2,4,6,8,10-dodecapentaenal.
[0032] The unsaturated aldehyde is preferably selected from the
group consisting of di-unsaturated aldehydes having 4 to 12
main-chain carbon atoms and having double bonds in positions 2 and
4, mono-unsaturated aldehydes having 4 to 9 main-chain carbon atoms
and having a double bond in position 2, and mono-unsaturated
aldehydes having 9 to 12 main-chain carbon atoms and having a
double bond in position 4. Examples of such an unsaturated aldehyde
include trans,trans-2,4-hexadienal, trans,trans-2,4-heptadienal,
trans,trans-2,4-nonadienal, trans,trans-2,4-decadienal,
trans,trans-2,4-dodecadienal, 2-butenal, trans-2-heptenal,
trans-2-octenal, trans-2-nonenal, trans-4-decenal, and
cis-4-decenal.
[0033] The CCK secretion-promoting agent of the present invention
preferably contains an unsaturated aldehyde having a double bond in
position 2. Examples of such an unsaturated aldehyde include
trans,trans-2,4-hexadienal, trans,trans-2,4-heptadienal,
trans,trans-2,4-nonadienal, trans,trans-2,4-decadienal,
trans,trans-2,4-dodecadienal, 2-butenal (crotonaldehyde),
trans-2-heptenal, trans-2-octenal, and trans-2-nonenal.
[0034] The CCK secretion-promoting agent containing an unsaturated
aldehyde having double bonds in positions 2 and 4 has a higher
CCK-secreting activity and is particularly preferred. Examples of
such an unsaturated aldehyde include trans,trans-2,4-hexadienal,
trans,trans-2,4-heptadienal, trans,trans-2,4-nonadienal,
trans-trans-2,4-decadienal, and trans,trans-2,4-dodecadienal.
[0035] The unsaturated aldehyde is preferably in a trans
conformation.
[0036] The CCK secretion-promoting agent preferably contains an
unsaturated aldehyde having two double bonds and an unsaturated
aldehyde having one double bond or contains two different
unsaturated aldehydes having two double bonds. It has been revealed
that the combination of two or more different unsaturated aldehydes
synergistically promotes the activity of secreting cholecystokinin.
The unsaturated aldehyde having only one double bond preferably has
eight or less carbon atoms and is more preferably trans-2-octenal.
The unsaturated aldehyde having two double bonds is preferably
trans,trans-2,4-hexadienal, trans,trans-2,4-heptadienal, or
trans,trans-2,4-decadienal. In the CCK secretion-promoting agent
containing two different unsaturated aldehydes having two double
bonds, one of the unsaturated aldehydes is preferably
trans,trans-2,4-heptadienal.
[0037] The CCK secretion-promoting agent of the present invention
induces, for example, a delay in the excretion of gastric contents,
a reduction in food intake (appetite), a decrease in the sensation
of hunger, or an increase in the feeling of fullness in animals
including human beings, as in the conventional
cholecystokinin-secreting activity. Accordingly, examples of the
use of the CCK secretion-promoting agent of the present invention
include appetite suppressants, hyperphagia preventive or
therapeutic agents, and obesity preventive or therapeutic agents
(hereinafter, referred to as appetite suppressant, etc.).
[0038] The CCK secretion-promoting agent, the appetite suppressant,
etc. of the present invention can be provided as drugs or
functional foods. The content of the unsaturated aldehyde in the
CCK secretion-promoting agent or the appetite suppressant, etc. is
usually 0.005% to 60% by weight, preferably 0.05% to 30% by
weight.
[0039] The CCK secretion-promoting agent and other agents of the
present invention can contain pharmacologically or bromatologically
acceptable other additives within a range that does not impair the
effects of the present invention, in addition to the unsaturated
aldehyde as an active ingredient. Examples of such additives
include excipients such as corn starch, crystalline cellulose, and
lactose; disintegrators such as starch, sodium alginate, gelatin,
calcium carbonate, and calcium citrate; binding agents such as
methyl cellulose and its salts, ethyl cellulose, gum arabic, and
gelatin; lubricants such as talc, magnesium stearate, polyethylene
glycol, and hydrogenated vegetable oils; xanthine derivatives; pH
adjusters; cooling agents; suspending agents; viscous agents;
solubilizers; antioxidants; coating agents; plasticizers;
surfactants; water; alcohols; water-soluble polymers; sweeteners
such as fructose, glucose, and sorbitol; corrigents; acidifiers
such as citric acid; flavoring agents; coloring agents; vitamins;
minerals; and lipids.
[0040] The CCK secretion-promoting agent and other agents of the
present invention may be in any form. Examples of the form include
solid formulations such as powders, granules, capsules, pills,
tablets, chewable tablets, and drops; and liquid formulations such
as drinkable preparations, liquid agents, suspensions, emulsions,
syrups, and dry syrups.
[0041] The CCK secretion-promoting agent and other agents of the
present invention may be administered in any route. In the case of
a drug, the drug may be administered orally or parenterally (e.g.,
intravenous injection, intramuscular injection, subcutaneous
injection, intraperitoneal administration, rectal administration,
or transdermal administration).
[0042] Since the mechanism of the action of CCK is a delay of the
transfer of gastric contents to the duodenum by closing the pylorus
of the stomach, the CCK secretion-promoting agent and other agents
of the present invention are desirably ingested before foods reach
the duodenum. As shown by the cell test described below, since the
CCK secretion-promoting agent produces the effect within 60 minutes
after the ingestion, specifically, the CCK secretion-promoting
agent and other agents of the present invention is preferably
ingested before or during the meal.
[0043] In the case of using the CCK secretion-promoting agent and
other agents of the present invention as drugs, the dosage varies
depending on the age, body weight, and anamnesis (e.g., obesity) of
a patient to whom the agent is administered. Specifically, the
daily dosage of the unsaturated aldehyde is usually 0.5 to 10 mg/kg
of body weight, preferably 1 to 5 mg/kg of body weight, for an
adult.
[0044] In the case of using the CCK secretion-promoting agent and
other agents of the present invention as functional foods, the
ingestion amount varies depending on the age, body weight, and
anamnesis (e.g., obesity) of a subject who ingests the agent.
Specifically, the daily ingestion of the unsaturated aldehyde is
usually 0.5 to 10 mg/kg of body weight, preferably 1 to 5 mg/kg of
body weight, for an adult.
[0045] The present invention also provides an appetite-suppressing
food product containing the CCK secretion-promoting agent. Examples
of the food include feeds for pets. A food containing the CCK
secretion-promoting agent of the present invention can suppress
food ingestion. Examples of the food include, but not limited to,
common processed foods and drinks, for example, side dishes such as
salads, fried foods, tofu, and konnyaku; soups; bread, rice, and
noodles; confectionery such as cookies, muffins, cakes, chips,
snack confectionery, chocolates, jellies, puddings, chewing gum,
and candies; dairy products such as yogurt and milk; fish meat
pastes such as ham, sausages, and boiled fish pastes; drinks such
as coffee, juices, and sports beverages; and flavorings such as
dressings, soy sauce, and sauces.
[0046] The amount of the unsaturated aldehyde in an
appetite-suppressing food product containing the CCK
secretion-promoting agent is preferably 0.001% to 0.03% by weight,
preferably 0.003% to 0.015% by weight.
EXAMPLES
[0047] The present invention will now be described in more detail
by means of Examples according to the present invention and
Comparative Examples, but is not limited to the following
examples.
Examples 1 to 12
1-1. Preparation of Test Material
[0048] As shown in Table 1, aldehydes having 3 to 13 main-chain
carbon atoms and having 0 to 2 double bonds in the main chain,
ketones, alcohols, fatty acids, and hydrocarbons were prepared. As
a condition for maximizing the secretion amount of CCK in a cell
test system, 70 mM KCl (positive control) causing hormone release
by depolarization was prepared.
[0049] Since the number of materials that can be tested at once is
restricted, the test materials were divided into five groups as
shown in Table 1. In order to solve the discontinuity between
experimental groups, every group included
trans,trans-2,4-decadienal. Each test material was tested three
times, and the average value of the results was calculated.
TABLE-US-00001 TABLE 1 Experi- mental group Test material 1
trans,trans-2,4-hexadienal, trans,trans-2,4-heptadienal,
trans,trans-2,4-nonadienal, trans,trans-2,4-decadienal,
trans-2-butenal (crotonaldehyde), trans-2-heptenal, butanal
(butyraldehyde), and nonanal 2 trans,trans-2,4-decadienal,
trans,trans-2,4-dodecadienal, trans-4- decenal, cis-4-decenal,
decanal, undecanal, dodecanal, tridecanal, trans-2-decenal,
cis-4-heptenal, and cis-7-decenal 3 trans,trans-2,4-decadienal,
trans-2-nonenal, octanal, trans-2- undecenal, trans-2-dodecenal,
cis-6-nonenal, and trans-10- undecenal 4
trans,trans-2,4-decadienal, trans-2-octenal, cis-4-decenal, acrylic
acid, 2-propen-1-ol, trans-2-octen-1-ol, cis-4-decen-1-ol, and
trans-2-octenoic acid 5 trans,trans-2,4-decadienal, octanal,
decanal, dodecanal, tridecanal, 1-octanol, n-decyl alcohol, dodecyl
alcohol, n-tridecyl alcohol, trans-3-decen-2-one, n-octanoic acid,
decanoic acid, lauric acid, n-tridecanoic acid, decane, cis-2
-decene, and trans,trans-1,9-decadiene
1-2. Process of Preparing Evaluation Solution
[0050] The test materials were each dissolved in ethanol, and each
solution was diluted with Hepes buffer (140 mM NaCl, 4.5 mM KCl, 20
mM Hepes, 1.2 mM CaCl.sub.2, 1.2 mM MgCl.sub.2, and 10 mM
D-glucose, pH 7.4) to give a 0.1 vol % ethanol solution, which was
used as the evaluation solution.
1-3. Confirmation Test of CCK Secretion-Stimulating Activity
[0051] Mouse intestinal CCK-producing cell line STC-1 was cultured
in a 48-well plate for two to three days until subconfluent in a
Dulbecco's modified Eagle's medium containing 10% bovine fetal
serum at 37.degree. C. in the presence of 5% CO.sub.2. After the
wells were washed with Hepes buffer, 100 .mu.L of the evaluation
solution containing 100 .mu.M of a test material was added to a
well, followed by incubation at 37.degree. C. for 60 minutes. The
supernatant was collected and was centrifuged (800.times.g, 5 mM,
4.degree. C.) to precipitate the exfoliated cells. The resulting
supernatant (80 .mu.L) was collected and was cryopreserved. The
concentration of CCK in the supernatant was measured with a
commercially available enzyme immunoassay kit (manufactured by
Phoenix Pharmaceuticals, Inc.).
[0052] The secretion amount of CCK varied from 10 to 30 pM among
the tests of a vehicle (i.e., a blank). The secretion amounts of
CCK in the tests for test materials (100 .mu.M) of Examples 1 to 12
reached 30 to 60 pM and were always two to three times that by the
vehicle, whereas the secretion amounts of CCK in the tests for test
materials of Comparative Examples were equivalent to that by the
vehicle or lower than that by the vehicle in some cases. Thus, it
was demonstrated that acrylic acid and the specific unsaturated
aldehydes used in Examples have a CCK secretion-stimulating
activity.
[0053] In order to investigate the change in the secretion amount
of CCK by addition of a test material to the cells in a general
condition, a value obtained by subtracting the measured value of
the vehicle from the measured value of a sample was used as the
true value, and the score of the CCK secretion-stimulating activity
was determined by dividing the true value by the value of 70 mM KCl
as a control. The variance was analyzed for each experimental
group. The difference in variance between an experimental group and
the vehicle was examined for determining whether there was a
significant difference (P<0.05). The results are shown in Tables
2A and 2B.
TABLE-US-00002 TABLE 2A Number Double bond CCK secretion-promoting
activity of Position Significant difference from vehicle Name of
carbon Position Position Struc- Group Group Group Group Group
ingredient Class atoms Number 2 4 Others ture Score 1 2 3 4 5
Control 70 mM KCl -- -- -- -- -- -- -- 1 Example 1 trans,trans-2,4-
aldehyde 6 2 .smallcircle. .smallcircle. trans 0.840 * hexadienal
Example 2 trans,trans-2,4- aldehyde 7 2 .smallcircle. .smallcircle.
trans 0.885 * heptadienal Example 3 trans,trans-2,4- aldehyde 9 2
.smallcircle. .smallcircle. trans 0.975 * nonadienal Example 4
trans,trans-2,4- aldehyde 10 2 .smallcircle. .smallcircle. trans
1.088 * * * * * decadienal Example 5 trans,trans-2,4- aldehyde 12 2
.smallcircle. .smallcircle. trans 0.720 * dodecadienal Example 6
trans-2-butenal aldehyde 4 1 .smallcircle. trans 0.644 *
(crotonalde- hyde) Example 7 trans-2-heptenal aldehyde 7 1
.smallcircle. trans 0.624 * Example 8 trans-2-octenal aldehyde 8 1
.smallcircle. trans 0.698 * Example 9 trans-2-nonenal aldehyde 9 1
.smallcircle. trans 0.491 -- Example 10 trans-4-decenal aldehyde 10
1 .smallcircle. trans 0.441 -- Example 11 cis-4-decenal aldehyde 10
1 .smallcircle. cis 0.377 -- -- Example 12 acrylic acid fatty acid
3 1 .smallcircle. trans 0.957 * *: A significant difference (P <
0.05) was found --: No significant difference was found
TABLE-US-00003 TABLE 2B Number Double bond CCK secretion-promoting
activity of Position Significant difference from vehicle Name of
carbon Position Position Struc- Group Group Group Group Group
ingredient Class atoms Number 2 4 Others ture Score 1 2 3 4 5
Control 70 mM KCl -- -- -- -- -- -- -- 1 Comparative butanal
aldehyde 4 0 0.017 -- Example 1 (butyral- dehyde) Comparative
octanal aldehyde 8 0 -0.004 -- -- Example 2 Comparative nonanal
aldehyde 9 0 -0.003 -- Example 3 Comparative decanal aldehyde 10 0
0.042 -- -- Example 4 Comparative undecanal aldehyde 11 0 0.189 --
Example 5 Comparative dodecanal aldehyde 12 0 -0.093 -- -- Example
6 Comparative tridecanal aldehyde 13 0 0.049 -- -- Example 7
Comparative trans-2- aldehyde 10 1 .smallcircle. trans 0.046 --
Example 8 decenal Comparative trans-2- aldehyde 11 1 .smallcircle.
trans -0.137 -- Example 9 undecenal Comparative trans-2- aldehyde
12 1 .smallcircle. trans -0.319 -- Example 10 dodecenal Comparative
cis-4- aldehyde 8 1 .smallcircle. cis -0.001 -- Example 11 heptenal
Comparative cis-6- aldehyde 9 1 position cis -0.648 -- Example 12
nonenal 6 Comparative cis-7- aldehyde 10 1 position cis 0.274 --
Example 13 decenal 7 Comparative trans-10- aldehyde 11 1 position
trans -0.458 -- Example 14 undecenal 10 Comparative 2-propen-
alcohol 3 1 .smallcircle. 0.557 -- Example 15 1-ol Comparative
trans-2-octen- alcohol 8 1 .smallcircle. trans 0.321 -- Example 16
1-ol Comparative cis-4-decen- alcohol 10 1 .smallcircle. cis -0.171
-- Example 17 1-ol Comparative l-octanol alcohol 8 0 -0.134 --
Example 18 Comparative n-decyl alcohol 10 0 0.070 -- Example 19
alcohol Comparative dodecyl alcohol 12 0 -0.304 -- Example 20
alcohol Comparative n-tridecyl alcohol 13 0 -0.225 -- Example 21
alcohol Comparative trans-3-decen- ketone 10 1 position trans 0.169
-- Example 22 2-one 3 Comparative trans-2- fatty 8 1 .smallcircle.
trans 0.146 -- Example 23 octenoic acid acid Comparative n-octanoic
fatty 8 0 -0.140 -- Example 24 acid acid Comparative decanoic fatty
10 0 -0.207 -- Example 25 acid acid Comparative lauric fatty 12 0
-0.225 -- Example 26 acid acid Comparative n-tridecanoic fatty 13 0
-0.234 -- Example 27 acid acid Comparative decane hydro- 10 0
-0.224 -- Example 28 carbon Comparative cis-2- hydro- 10 1
.smallcircle. cis -0.208 -- Example 29 decene carbon Comparative
trans,trans- hydro- 10 2 positions trans -0.211 -- Example 30
1,9-decadiene carbon 1, 9 *: A significant difference (P < 0.05)
was found --: No significant difference was found
[0054] Tables 2A and 2B revealed the following: A CCK
secretion-stimulating activity was observed in unsaturated
aldehydes of Examples 1 to 11 with significant differences. The
comparison between Examples 1 to 11 and Comparative Examples 15 to
30 demonstrates that alcohols, fatty acids excluding acrylic acid,
and hydrocarbons do not have a CCK secretion-stimulating activity,
whereas aldehydes have a CCK secretion-stimulating activity. The
comparison between Examples 1 to 11 and Comparative Examples 1 to 7
demonstrates that the aldehydes must be unsaturated aldehydes. The
comparison between Examples 1 to 11 and Comparative Examples 12 to
14 demonstrates that a double bond must be present in at least
position 2 or 4.
[0055] The comparison between Examples 6 to 9 and Comparative
Examples 8 to 10 demonstrates that an unsaturated aldehyde having a
double bond in only position 2 must have 9 or less main-chain
carbon atoms. The comparison between Examples 10 to 11 and
Comparative Example 11 demonstrates that an unsaturated aldehyde
having a double bond in only position 4 must have 9 or more
main-chain carbon atoms.
[0056] The comparison between Example 10 and Example 11
demonstrates that preferred unsaturated aldehydes are in trans
conformations. The superiority of the trans conformation is also
suggested in that the best results are obtained in Examples 1 to 5
all in trans conformations.
[0057] As shown in Examples 1 to 5, the excellent results regarding
the CCK secretion-stimulating activity are observed in unsaturated
aldehydes having double bonds in positions 2 and 4. In particular,
the CCK secretion-stimulating activity of
trans,trans-2,4-decadienal is the highest.
[0058] It has been confirmed in a cell test system of a
conventional technology that an ingredient (e.g., a degradation
product of (.beta.-conglycinin) showing a CCK-secreting activity
suppresses appetite. Accordingly, it is obvious that the CCK
secretion-promoting agent of the present invention also functions
as an appetite suppressant.
Examples 13 to 23
Synergistic Effect of CCK Secretion-Stimulating Activity
[0059] Combinations of two different unsaturated aldehydes shown in
Table 3 were evaluated for their CCK secretion-stimulating
activity. The evaluation was performed as in Example 1 except that
the concentrations of the two different test materials were each 50
.mu.M (a half of the concentration in the test described in
paragraph 1-3). The results are shown in Table 3.
TABLE-US-00004 TABLE 3 CCK secretion- promoting activity
Significant difference Ingredient 1 Ingredient 2 Score from vehicle
Control 70 mM KCl (100 .mu.M) 1 Example 4
trans,trans-2,4-decadienal (100 .mu.M) 1.088 * Example 13
trans,trans-2,4- trans,trans-2,4- 0.492 * decadienal hexadienal
Example 14 trans,trans-2,4- trans,trans-2,4- 0.795 * decadienal
heptadienal Example 15 trans,trans-2,4- trans-2-octenal 1.127 *
decadienal Example 16 trans,trans-2,4- trans-2-nonenal 0.594 --
decadienal Example 17 trans,trans-2,4- trans,trans-2,4- 0.934 *
hexadienal heptadienal Example 18 trans,trans-2,4- trans-2-octenal
1.190 * hexadienal Example 19 trans,trans-2,4- trans-2-nonenal
0.868 * hexadienal Example 20 trans,trans-2,4- trans-2-octenal
1.383 * heptadienal Example 21 trans-2-octenal trans-2-nonenal
0.479 -- * A significant difference (P < 0.05) was found --: No
significant difference was found
[0060] Table 3 demonstrates that the CCK-secreting activity is
promoted by a combination of an unsaturated aldehyde having two
double bonds and an unsaturated aldehyde having one double bond or
by a combination of two different unsaturated aldehydes having two
double bonds. It has been demonstrated that the CCK-secreting
activity is highly promoted, in particular, by a combination of an
unsaturated aldehyde having two double bonds and an unsaturated
aldehyde having eight or less carbon atoms and one double bond. It
has been demonstrated that the CCK-secreting activity is further
highly promoted by a combination of an unsaturated aldehyde having
double bonds in positions 2 and 4 and trans-2-octenal having a
double bond in position 2.
* * * * *
References