U.S. patent application number 12/593438 was filed with the patent office on 2010-06-17 for medicinal composition, food or drink having effect of enhancing sympathetic nervous activity.
This patent application is currently assigned to SUNTORY HOLDINGS LIMITED. Invention is credited to Yoshinori Beppu, Hajime Komura, Katsuya Nagai, Tomohiro Rogi, Nobuo Tsuruoka.
Application Number | 20100150890 12/593438 |
Document ID | / |
Family ID | 37023848 |
Filed Date | 2010-06-17 |
United States Patent
Application |
20100150890 |
Kind Code |
A1 |
Beppu; Yoshinori ; et
al. |
June 17, 2010 |
MEDICINAL COMPOSITION, FOOD OR DRINK HAVING EFFECT OF ENHANCING
SYMPATHETIC NERVOUS ACTIVITY
Abstract
It is intended to provide a safe composition for preventing or
treating obesity or related complications which has an autonomic
nervous control effect, in particular, an effect of enhancing
sympathetic nervous activity and an effect of promoting energy
metabolism. Thus, a medicinal composition, a food or a drink
containing 2,3-dihydro-3,5-dihydroxy-6-methyl-4H-pyran-4-one and
having an autonomic nervous control effect and an effect of
promoting energy metabolism is provided.
Inventors: |
Beppu; Yoshinori; (Osaka,
JP) ; Tsuruoka; Nobuo; (Osaka, JP) ; Rogi;
Tomohiro; (Osaka, JP) ; Komura; Hajime;
(Osaka, JP) ; Nagai; Katsuya; (Osaka, JP) |
Correspondence
Address: |
DRINKER BIDDLE & REATH (DC)
1500 K STREET, N.W., SUITE 1100
WASHINGTON
DC
20005-1209
US
|
Assignee: |
SUNTORY HOLDINGS LIMITED
Osaka-shi
JP
|
Family ID: |
37023848 |
Appl. No.: |
12/593438 |
Filed: |
March 28, 2008 |
PCT Filed: |
March 28, 2008 |
PCT NO: |
PCT/JP2008/056075 |
371 Date: |
September 28, 2009 |
Current U.S.
Class: |
424/93.42 ;
424/93.1; 424/93.4; 424/93.45; 424/93.48; 514/460 |
Current CPC
Class: |
C12N 1/14 20130101; A23F
5/02 20130101 |
Class at
Publication: |
424/93.42 ;
514/460; 424/93.1; 424/93.4; 424/93.45; 424/93.48 |
International
Class: |
A61K 35/74 20060101
A61K035/74; A61K 31/35 20060101 A61K031/35; A61K 35/66 20060101
A61K035/66 |
Foreign Application Data
Date |
Code |
Application Number |
Mar 24, 2005 |
JP |
2005-086884 |
Claims
1. A medicinal composition, a food or a drink which contains a
clinically effective amount of
2,3-dihydro-3,5-dihydroxy-6-methyl-4H-pyran-4-one and a
pharmaceutically or nutritionally acceptable additive and has an
autonomic nervous control effect.
2. A medicinal composition, food or drink as claimed in claim 1
wherein the autonomic nervous control effect is an effect of
enhancing sympathetic nervous activity.
3. A medicinal composition, food or drink as claimed in claim 1
which has an effect of promoting energy metabolism.
4. A medicinal composition, food or drink as claimed in claim 1
which is to be used for treating a disease or condition relating to
autonomic nervous activity or energy metabolism.
5. A medicinal composition, food or drink as claimed in claim 4
wherein the disease or condition relating to autonomic nervous
activity or energy metabolism is obesity and/or related
complications.
6. A medicinal composition, food or drink as claimed in claim 1
which contains a microorganism capable of producing
2,3-dihydro-3,5-dihydroxy-6-methyl-4H-pyran-4-one or a processed
product thereof.
7. A medicinal composition, food or drink as claimed in claim 1
which contains a culture supernatant of a microorganism capable of
producing 2,3-dihydro-3,5-dihydroxy-6-methyl-4H-pyran-4-one or a
processed product thereof.
8. A medicinal composition, food or drink as claimed in claim 6
wherein the microorganism is a lactic acid bacterium.
9. A medicinal composition, food or drink as claimed in claim 8
wherein the lactic acid bacterium is one belonging to the genus
Streptococcus, Lactobacillus, Leuconostoc, Pediococcus,
Bifidobacterium, Tetragenococcus, Weissella, Enterococcus,
Melisscoccus, Lactococcus, Carnobacterium, Vagococcus, Atopobium,
Lactosphaera, Oenococcus, Abiotrophia, Paralactobacillus,
Granulicatella, Atopobactor, Alkalibacterium or Olsenella.
10. A medicinal composition, food or drink as claimed in claim 6
wherein the microorganism belongs to Lactobacillus plantarum
(preferably Lactobacillus plantarum SAM 2446 strain (FERM
BP-10438)) or Lactobacillus brevis (preferably Lactobacillus brevis
SAM 2447 strain (FERM BP-10439)).
11. A method of producing a medicinal composition, food or drink as
claimed in claim 1 which comprises the step of culturing a
microorganism capable of producing
2,3-dihydro-3,5-dihydroxy-6-methyl-4H-pyran-4-one in a medium.
Description
TECHNICAL FIELD
[0001] The present invention relates to a medicinal composition, a
food or a drink which contains
2,3-dihydro-3,5-dihydroxy-6-methyl-4H-pyran-4-one and has an
autonomic nervous control effect. Further, it relates to the
medicinal composition, food or drink as described above which has
an effect of enhancing sympathetic nervous activity and an effect
of promoting energy metabolism and is to be used for treating a
disease or condition relating to the autonomic nervous activity or
energy metabolism, in particular, obesity and/or related
complications.
BACKGROUND ART
[0002] To maintain the homeostasis, a living body has the autonomic
nervous system, the endocrine system and the immune system. Among
these systems, the autonomic nervous system is relatively free from
the cerebral control and acts automatically without being
consciously controlled, which is the origin of the name. It mainly
controls visceral functions. The autonomic nervous system comprises
the sympathetic nervous system and the parasympathetic nervous
system and is controlled based on a balance between these two
systems. When the body is in action, the sympathetic nervous
activity become predominant and the whole body is in a state of
tension. On the contrary, when the parasympathetic nervous activity
become predominant, the tension is loosened and the body is relaxed
(Non-Patent Document 1). Sympathetic hyperactivity causes pupil
dilation, tachycardia, a rise in blood pressure, substance
metabolism, hyperglycemia and so on. Parasympathetic hyperactivity
causes activation of the digestion and absorption system, secretion
of sweat and saliva and so on. It is also known that the autonomic
nervous activity relate to control of the white adipose tissue and
the brown adipose tissue functions. Namely, the white adipose
tissue function is controlled by the sympathetic nerve and hormones
in the blood, while the brown adipose tissue function is controlled
mainly by the sympathetic nerve (Non-Patent Document 2).
[0003] On the other hand, obesity has become a serious health
problem not only in developed countries but also in developing
countries, so that antiobesity measures are required worldwide. In
people having BMI (body mass index; a standard defining obesity) of
25 or higher, prevalences of various diseases such as hypertension,
hyperglycemia and hyperglyceridemia show linear increases with an
increase in BMI (Non-Patent Document 3). Also, it has been
clarified that many pathological conditions induced by obesity
relate not only to a degree of obesity but also to visceral fat
accumulation.
[0004] Although antiobesity drugs have been developed and used
around the world under these circumstances, it is also known that
these drugs exert side effects. As a typical example thereof,
sibutramine (Meridia manufactured by Knoll, etc.) has a possible
risk of inducing an increase in heart rate or a rise in blood
pressure, which makes it difficult to administer this drug to
patients with symptoms frequently occurring in obese persons such
as hypertension, cardiac angina, congestive heart failure or
cerebral stroke. Orlistat (Xenical manufactured by Roche) sometimes
shows serious side effects of diarrhea and steatorrhea.
[0005] In Japan, as antiobesity drugs, mazindol
(Sanorex)(Non-Patent Document 4), which is reported as having an
antiobesity effect via the activation of the brown adipose tissue,
and Farng Feng Tong San (bohutusyo-san) have been mainly employed.
Farng Feng Tong San is composed of 18 kinds of crude drugs
including ephedra herb which contains 1-ephedrine and
d-pseudophedrine and enhances the release of noradrenaline from the
sympathetic nervous ending to thereby activate .beta.-adrenaline
receptor in the brown adipose tissue (Non-Patent Document 5). It is
reported that these drugs exert side effects too. That is, it has
been confirmed that mazindol causes thirst, constipation, nausea
and sleep disorders while Farng Feng Tong San causes hepatic
disorders.
[0006] Paying attention to therapeutic effects of lactic acid
bacteria, in particular, there have been disclosed an antiobesity
effect of a fermented product obtained by fermenting carrot root by
using lactic acid bacteria (Patent Document 1), an effect of
preventing visceral fat accumulation achieved by a lactic acid
bacterium cultured product (Patent Document 2) and so on. However,
no specific action on the white adipose tissue and the brown
adipose tissue has been verified so far. Paying attention to health
improvement using the autonomic nervous control effect of lactic
acid bacteria, there has been merely reported an effect of
alleviating constipation (Patent Document 3) but no effect on
obesity has been disclosed so far.
[0007] On the other hand, concerning
2,3-dihydro-3,5-dihydroxy-6-methyl-4H-pyran-4-one which is
contained in various foods and can be synthesized by a plurality of
known methods, there have been known an inhibitory effect of
tyrosinase, an inhibitory effect of melanogenesis (Patent Document
4) and an antioxidant effect (Patent Document 5) alone.
Patent Document 1: JP-A-2004-201598
Patent Document 2: JP-A-2004-99539
Patent Document 3: JP-A-2004-155727
Patent Document 4: JP-A-2006-28077
Patent Document 5: JP-A-H04-1291 (JP No. 2843639)
Non-Patent Document 1: Toshio Hagihara, at al., Iwanami Koza,
Gendai Igaku no Kiso, vol. 4, Seitai no Chousetsu Shisutemu,
1999
[0008] Non-Patent Document 2: Nippon Rinsho, vol. 6, extra ed. 6,
288-294, 2003
Non-Patent Document 3: Himan Kenkyu 6:4-17, 2000
[0009] Non-Patent Document 4: Yoshida T., et al.: Clin. Exp.
Pharmacol. Physiol. 23:476-482, 1996 Non-Patent Document 5: Nippon
Rinsho, vol. 61, extra. ed. 6, 649-654, 2003
DISCLOSURE OF THE INVENTION
Problems that the Invention is to Solve
[0010] There are a large number of autonomic nervous disorders, and
of health disorders caused thereby. However, it has been a common
practice to separately treat these disorders, and no attempt has
been made so far to control the autonomic nervous activity per se
that are the fundamental cause for these disorders. Although each
symptom can be temporarily alleviated by such a symptomatic
treatment, there still remains a high risk of recurrence or of the
onset of another symptom caused by autonomic nervous disorders.
Thus, there have been required drugs, foods and drinks whereby the
autonomic nervous activity per se, i.e., the fundamental cause can
be controlled.
[0011] Moreover, preventives and remedies for obesity and related
complications that have been widely used hitherto exhibit side
effects. Thus, it is difficult to establish the desired antiobesity
effects, minimal side effects and high safety at the same time.
Furthermore, a process for synthesizing a drug involves additional
step(s) and a thus synthesized drug should be processed into a form
assuring easy intake thereof such as tablets, which brings about
another problem; high cost. Under these circumstances, it has been
required to utilize the antiobesity effect of a safe food material
or a food-origin component that has been taken by human beings for
a long time and it has been also strongly required to develop an
active substance which can be obtained from a natural material
occurring in nature via a simple process.
[0012] No remedy for obesity based on the promotion of energy
consumption has been developed so far (Nippon Rinsho, vol. 61,
extra ed. 6, 686-691, 2003). Since the relationship between
enlargement in adipose tissue and insulin resistance has been
clarified, it is expected that an energy consumption promoter
ensuring a qualitative change of enlarged adipose cells into
small-sized cells having normal functions would be usable as an
ideal remedy for obesity. When an antiobesity effect is to be
achieved by enhancing the sympathetic nervous activity of adipose
tissue from the above-described point of view, it is anticipated
that blood pressure would be raised. Thus, this phenomenon should
be avoided. Accordingly, it is ideal, if possible, that such an
antiobesity effect based on the promotion of energy consumption
having the characteristics as discussed above is provided with the
use of a safe food material or a food-origin component that has
been taken by human beings for a long time.
[0013] An object of the present invention is to provide a safe
composition for preventing or treating obesity and/or related
complications which have an autonomic nervous control effect; in
particular, an effect of enhancing sympathetic nervous activity
and, moreover, an effect of promoting energy metabolism.
Means for Solving the Problems
[0014] It has been confirmed that cells of a specific lactic acid
bacterium can reduce sympathetic nervous activity of the kidney and
enhance parasympathetic nervous activity of the stomach (Tanida M,
et al., Neuroscience Letters 389:109-114, 2005).
[0015] To achieve the object as described above, the present
inventors conducted intensive studies and isolated
2,3-dihydro-3,5-dihydroxy-6-methyl-4H-pyran-4-one from a culture
supernatant of a lactic acid bacterium with the use of the direct
effects on the rat autonomic nervous activity as an indication.
Then, they have found that this compound has an effect of rather
lowering blood pressure, though it has an effect of enhancing
sympathetic nervous activity of the kidney and that it has another
effect of enhancing sympathetic nervous activity of the white
adipose tissue and the brown adipose tissue. Moreover, they have
found that this compound has still another effect of promoting
energy metabolism, thus completing the present invention.
[0016] Accordingly, the present invention relates to a medicinal
composition, a food or a drink which contains a clinically
effective amount of
2,3-dihydro-3,5-dihydroxy-6-methyl-4H-pyran-4-one and a
pharmaceutically or nutritionally acceptable additive, and has an
autonomic nervous control effect.
[0017] The invention further relates to the medicinal composition,
food or drink as described above wherein the autonomic nervous
control effect is an effect of enhancing sympathetic nervous
activity.
[0018] The invention further relates to a medicinal composition, a
food or a drink, which has an effect of promoting energy
metabolism.
[0019] The invention further relates to the medicinal composition,
food or drink as described above which is to be used for treating a
disease or condition relating to autonomic nervous activity or
energy metabolism.
[0020] The invention further relates to the medicinal composition,
food or drink as described above wherein the disease or condition
relating to the autonomic nervous activity or energy metabolism is
obesity and/or related complications (for example,
diabetes/impaired glucose tolerance, hyperlipemia, hypertension,
hyperuricemia/gout, coronary artery diseases (heart infarction,
angina pectoris, etc.), brain infarction (brain thrombosis,
transient ischemic attack, etc.), sleep apnea syndrome, fatty
liver, orthopedic diseases (arthritis deformans, lumbar disease,
etc.), menstruation disorder and so on).
[0021] The invention further relates to the medicinal composition,
food or drink as described above which contains a microorganism
capable of producing
2,3-dihydro-3,5-dihydroxy-6-methyl-4H-pyran-4-one or a processed
product thereof.
[0022] The invention further relates to the medicinal composition,
food or drink as described above which contains a culture
supernatant of a microorganism capable of producing
2,3-dihydro-3,5-dihydroxy-6-methyl-4H-pyran-4-one or a processed
product thereof.
[0023] The invention further relates to the medicinal composition,
food or drink as described above wherein the microorganism is a
lactic acid bacterium.
[0024] The invention further relates to the medicinal composition,
food or drink as described above wherein the lactic acid bacterium
is one belonging to the genus Streptococcus, Lactobacillus,
Leuconostoc, Pediococcus, Bifidobacterium, Tetragenococcus,
Weissella, Enterococcus, Melisscoccus, Lactococcus, Carnobacterium,
Vagococcus, Atopobium, Lactosphaera, Oenococcus, Abiotrophia,
Paralactobacillus, Granulicatella, Atopobactor, Alkalibacterium or
Olsenella. The invention further relates to the medicinal
composition, food or drink as described above wherein the
microorganism belongs to Lactobacillus plantarum (preferably
Lactobacillus plantarum SAM 2446 strain (FERM ABP-10438)) or
Lactobacillus brevis (preferably Lactobacillus brevis SAM 2447
strain (FERM ABP-10439)).
[0025] The invention further relates to a method of producing the
medicinal composition, food or drink as described above which
comprises the step of culturing a microorganism capable of
producing 2,3-dihydro-3,5-dihydroxy-6-methyl-4H-pyran-4-one in a
medium.
[0026] The invention further relates to the medicinal composition,
food or drink as described above wherein the autonomic nervous
control effect is one or more effects (preferably all effects)
selected from the group consisting of an effect of enhancing
sympathetic nervous activity of the kidney, an effect of enhancing
sympathetic nervous activity of the adrenal gland, an effect of
reducing vagal (parasympathetic) nervous activity of the stomach,
an effect of enhancing sympathetic nervous activity of the white
adipose tissue and an effect of enhancing sympathetic nervous
activity of the brown adipose tissue.
[0027] The invention further relates to the medicinal composition,
food or drink as described above wherein autonomic nervous control
effect is not accompanied by a rise in blood pressure.
[0028] The invention further relates to the medicinal composition,
food or drink as described above which has an effect of promoting
energy metabolism and/or an effect of promoting fat burning.
[0029] The invention further relates to the medicinal composition
as described above which is to be orally administered.
[0030] The invention further relates to an autonomic modulator
which contains 2,3-dihydro-3,5-dihydroxy-6-methyl-4H-pyran-4-one as
the active ingredient.
[0031] The invention further relates to use of
2,3-dihydro-3,5-dihydroxy-6-methyl-4H-pyran-4-one in producing a
medicinal composition, a food or a drink for treating a disease or
condition relating to autonomic nervous activity.
[0032] The invention further relates to a method of treating a
disease relating to autonomic nervous activity which comprises
administering
2,3-dihydro-3,5-dihydroxy-6-methyl-4H-pyran-4-one.
[0033] The invention further relates to an energy metabolism
controlling agent which contains
2,3-dihydro-3,5-dihydroxy-6-methyl-4H-pyran-4-one as the active
ingredient.
[0034] The invention further relates to use of
2,3-dihydro-3,5-dihydroxy-6-methyl-4H-pyran-4-one in producing a
medicinal composition, a food or a drink for treating a disease or
condition relating to energy metabolism.
[0035] The invention further relates to an autonomic nervous
control method which comprises administering
2,3-dihydro-3,5-dihydroxy-6-methyl-4H-pyran-4-one.
[0036] The invention further relates to an energy metabolism
controlling method which comprises administering
2,3-dihydro-3,5-dihydroxy-6-methyl-4H-pyran-4-one.
[0037] 2,3-Dihydro-3,5-dihydroxy-6-methyl-4H-pyran-4-one
(hereinafter sometimes referred to simply as "DDMP") to be used in
the present invention is represented by the following formula.
##STR00001##
[0038] This compound is obtained in, for example, Example 1 as will
be described hereinafter. A medicinal composition, a food or a
drink (hereinafter sometimes referred to simply as "composition")
containing DDMP has an autonomic nervous control effect; in
particular, an effect of enhancing sympathetic nervous activity.
Further, it has an effect of promoting energy metabolism, which
makes it particularly useful in treating obesity and/or related
complications.
[0039] It is known that DDMP, which is a compound analogous to
maltol known as an aroma component, is contained in various foods
and so on. For example, it is contained in various kinds of foods
and so on, such as propolis, oak barrels, maguey, wine, brandy,
milk, cookies, birch sap, popcorn, strawberry jam, blackstrap
molasses, brown sugar and so on. DDMP increases in the course of
aging of wine (Journal International des Sciences de la Vigne et du
Vin 32:99-110, 1998) and brandy (Priklanda Biokhmiyai
Mikrobiologiya 15:902-908, 1979). On the other hand, it is formed
by a Maillard reaction in an oak barrel (Cutzach Izabelle et al.:
Journal of Agricultural and Food Chemistry 45:2217-2224, 1997),
maguey (Lopez, M. G., et al.: Frontiers of Flavour Science:
523-526, 2000) and milk (Pischetsrieder, Monica et al.: Food
Research and Technology, 208:172-177, 1999).
[0040] Therefore, DDMP and the DDMP-containing composition as
described above can be obtained from foods, plants, etc. containing
DDMP or materials in which DDMP has been formed by aging, heating,
a Maillard reaction or the like. In forming DDMP, an optional
procedure well known to a person skilled in the art may be
conducted so as to increase the yield. To efficiently form the
target compound by the Maillard reaction, for example, it is a
common practice to add ascorbic acid or another substance capable
of participating in the Maillard reaction.
[0041] As will be described hereinafter in Examples, the present
inventors have also found that DDMP is contained in a culture
supernatant of a lactic acid bacterium (Lactobacillus plantarum SAM
2446 strain (FERM ABP-10438)).
[0042] To separate DDMP from the above-described material, culture
supernatant, etc., the starting material may be fractionated as
such. Alternatively, it may be concentrated by freeze drying,
liquid-liquid partition using an organic solvent or the like prior
to the fractionation. The subsequent fractionation procedure may be
carried out by, if desired, conducting of crude fractionation by
ultrafiltration or the like followed by a treatment with the use of
a reversed phase column, etc. The liquid-liquid partition,
ultrafiltration, reversed phase column treatment, etc. as described
above can be conducted by methods commonly employed by a person
skilled in the art.
[0043] It is also possible to obtain DDMP by synthesis. Examples of
the method of synthesizing DDMP include a method which comprises
heating glucose with .beta.-alanine (Nishibori S., et al.: Journal
of Agricultural and Food Chemistry 41:2374-2377, 1993), a method
which comprises heating sucrose with wine yeast (Mishiev, P. J., et
al.: Prikladnyaba Biokhimiya i Mikrobiologiya 17:307-311, 1981), a
method which comprises heating glucose with piperidine (Chen
Yongkuan et al.: Huaxue Yanjiu Yu Yingyong 15:45-48, 2003), a
method which comprises using glucose, piperidine and thioglycolic
acid (Isabelle Cutzach et al.: J. Agric. Food Chem. 47:1663-67,
1999), a method which comprises heating a saccharide composition
during hydrolysis of starch with L-arginine and glutamic acid, or a
saccharide composition during hydrolysis of starch with glutamic
acid (Song Wendong et al.: Fenix Huaxue 29:620, 2001) and so on,
though the synthesis method is not particularly restricted.
[0044] The formation of DDMP by such a method can be confirmed by,
for example, MS spectrometry or NMR spectrometry.
[0045] DDMP contained in the composition according to the present
invention may be used in the form of a pharmaceutically acceptable
salt such as hydrochloride or in the form of a salt (a prodrug)
that is converted into DDMP in vivo. Also, use can be made of a
D-compound, an L-compound or a racemate.
[0046] It has been reported that DDMP, which is an approved food
additive perfume having a high safety, has an inhibitory effect of
melanogenesis (Patent Document 4) and an antioxidant effect (Patent
Document 5). However, it has never been reported so far that DDMP
has an effect on the autonomic nerve, an effect on the sympathetic
nerve, an effect of promoting energy metabolism, or an effect
targeting the treatment of obesity and/or related
complications.
[0047] The present invention also provides the medicinal
composition, food or drink as described above which contains a
microorganism capable of producing DDMP or a processed product of
the same, or a culture supernatant of this microorganism or a
processed product of the same.
[0048] The expression "capable of producing DDMP" as used herein
means capable of producing DDMP when cultured under effective
conditions. Such effective conditions can be appropriately
determined by a person skilled in the art. The capability of
producing DDMP of a microorganism can be confirmed by analyzing the
microorganism (including a microorganism per se, dried cells of a
microorganism, a culture medium of a microorganism, an extract of a
microorganism and so on) by using a commonly employed procedure
such as LC, MS or NMR to examine the presence of DDMP, etc. A
microorganism capable of producing DDMP in the case of
appropriately controlling the culture conditions (the culture
composition, culture temperature, culture pH, culture density,
etc.) also falls within the scope of the microorganism capable of
producing DDMP.
[0049] The microorganism as described above may be collected from a
natural source. Moreover, a variant and/or a recombinant designed
as being capable of producing DDMP are also included in the scope
of the present invention. A variant and/or a recombinant designed
as showing an elevated DDMP productivity compared with a wild
strain when cultured in a culture medium of the same composition
also fall within the scope.
[0050] The microorganism capable of producing DDMP may be, for
example, a lactic acid bacterium, a yeast, a hay bacillus or the
like. Examples of the lactic acid bacterium include microorganisms
belonging to the genus Streptococcus, Lactobacillus, Leuconostoc,
Pediococcus, Bifidobacterium, Tetragenococcus, Weissella,
Enterococcus, Melisscoccus, Lactococcus, Carnobacterium,
Vagococcus, Atopobium, Lactosphaera, Oenococcus, Abiotrophia,
Paralactobacillus, Granulicatella, Atopobactor, Alkalibacterium or
Olsenella; examples of the yeast include microorganisms belonging
to the genus Candida or Saccharomyces; and examples of the hay
bacillus include microorganisms belonging to Bacillus subtilis.
[0051] Particularly preferable microorganisms are Lactobacillus
plantarum (more specifically Lactobacillus plantarum SAM 2446
strain (FERM ABP-10438)) and Lactobacillus brevis (more
specifically Lactobacillus brevis SAM 2447 strain (FERM
ABP-10439)).
[0052] The present invention further provides a method of producing
the medicinal composition, food or drink as described above which
comprises the step of culturing the above-described microorganism
capable of producing DDMP in a medium. The microorganism can be
cultured by inoculation of an appropriate medium with the
microorganism and employing of a culture procedure that is well
known to a person skilled in the art, depending on the kind of the
microorganism.
[0053] Next, the culture in the case where the microorganism is a
lactic acid bacterium will be briefly described by way of example.
As the medium, use can be made of, for example, an agar medium
and/or a liquid medium. To the medium, a carbon source and a
nitrogen source well known to a person skilled in the art are
optionally added, each at a desired concentration. If necessary,
trace nutrients such as an inorganic ion and a vitamin may be
further added. More conveniently, use can be made of a marketed
medium such as MRS medium to which additives may be optionally
added. After preparation of the medium, it is adjusted to pH 6.0 to
7.0 with the use of an appropriate acid or base and then sterilized
using an autoclave or the like.
[0054] Subsequently, the medium is inoculated with the lactic acid
bacterium. Then, the microorganism can be proliferated in the
medium by shaken culture, static culture, industrial-scaled culture
in a tank, solid culture in a solid medium such as an agar medium,
etc. while the culture temperature is controlled to 10.degree. C.
to 45.degree. C. usually for 1 to 2 days. The culture conditions
vary depending on the microorganism employed. In the case of using
Lactobacillus plantarum, for example, static culture can be
conducted in MRS medium of around pH 6.5 at around 37.degree. C.
for 1 day. The microorganism thus cultured is centrifuged if
desired, and then filtered if necessary. Thus, a culture
supernatant can be obtained.
[0055] As will be shown in Examples hereinafter, DDMP has an effect
of enhancing the rat kidney sympathetic nervous activity without
causing a rise in blood pressure when transduodenally administered
to rats. Further, it has an effect of reducing vagal
(parasympathetic) nervous activity of the stomach, an effect of
enhancing sympathetic nervous activity of the interscapular brown
adipose tissue and an effect of enhancing sympathetic nervous
activity of the epididymal white adipose tissue in rats. That is to
say, DDMP is intestinal canal-inhibitive, enhances sympathetic
nervous activity (the kidney and the adrenal gland), reduces vagal
(parasympathetic) nervous activity of the stomach and further
enhances sympathetic nervous activity of the white adipose tissue
and the brown adipose tissue without causing a rise in blood
pressure.
[0056] As will be shown in Examples hereinafter, DDMP caused an
increase in the rat oxygen consumption when forcibly orally
administered into the stomach of rats, which indicates that DDMP
has an effect of promoting energy metabolism.
[0057] As methods for evaluating autonomic nervous activity, there
can be enumerated biophysical measurement methods using, for
example, an electrocardiograph, a sphygmomanometer, a galvanic skin
reflex meter or a pupil meter and biochemical methods of measuring
blood catecholamine concentration, etc. Although analysis on heart
rate change using an electrocardiograph is a commonly employed
method, the present inventors evaluated the effects of DDMP on
autonomic nervous activity by a method using rats, as will be shown
in Examples hereinafter, whereby the effects of the administered
substance on the autonomic nervous activity can be directly
evaluated. This method is characterized in that the effects of the
administered substance on the autonomic nervous activity
controlling individual organs, tissues and so on can be separately
examined. Moreover, this method has a merit that autonomic nervous
control functions of different substances can be successively
evaluated so long as the test animal survives.
[0058] The following table shows organs, hormones and vital actions
seemingly participating in autonomic nervous activity, possible
effects of the autonomic nerve thereon, and examples of the
expected effects.
TABLE-US-00001 TABLE 1 Sympathomimetic Parasympathomimetic Pancreas
Insulin Decrease Increase Glucagon Increase Decrease Adrenal Blood
glucose Increase Decrease gland Blood pressure Increase Decrease
Liver Glycogen Decomposition Synthesis Sugar Gluconeogenesis
Glycolysis .fwdarw.Sugar synthesis enhancement .fwdarw.Sugar use
promotion Kidney Blood Increase Decrease pressure Stomach
Digestion/ Decrease Promotion absorption .fwdarw.Loss of appetite
.fwdarw.Appetite (antiobesity) enhancement Skin Blood flow rate
Decrease Increase .fwdarw.Moisturization enhancement (cosmetic)
Immunity Decrease Increase .fwdarw.NK activity rise Pupil
Enlargement Contraction Heart rate Increase Decrease Cardio-
Enhancement Regulation vascular (facial pallor) (facial flush)
system Sweat and Secretion Secretion enhancement saliva
regulation
[0059] As the above table clearly shows, the autonomic nervous
activity closely affect various controlling mechanisms in vivo, for
example, blood sugar control, blood pressure control, hormone
secretion control, sugar metabolism control, gastric juice
secretion control, blood flow rate control, body temperature
control and so on. When the sympathetic nervous activity are
enhanced, for example, there arise fat decomposition in the white
adipose tissue and heat production enhancement in the brown adipose
tissue (Non-Patent Document 2), sugar synthesis due to
gluconeogenesis in the liver, and a decrease in the
digestion/absorption in the stomach. When the parasympathetic
nervous activity are enhanced, there arise promotion of sugar use
due to glycolysis enhancement in the liver, appetite enhancement in
the stomach, moisturization enhancement due to an increase in blood
flow rate in the skin and immunopotentiation due to a rise in NK
activity in the immunity.
[0060] When particular attention is paid to antiobesity measures,
visceral fat, among the fats relating to obesity, has a higher
metabolic activity than subcutaneous fat. Therefore, it is known
that visceral fat is quickly synthesized at hyperalimentation and
vigorously synthesized and decomposed during exercise or in cases
of malnutrition. Accordingly, it is considered that fat synthesis
and decomposition are both vigorously conducted in an individual
with visceral fat accumulation and the resulting free fatty acids
that are the metabolites are released in a large amount. In
addition, these free fatty acids released from the visceral fat
would flow directly into the liver via the portal vein. The
excessive inflow of the free fatty acids regulates the insulin
clearance in the liver and potentiates gluconeogenesis, thereby
elevating blood sugar level. As a result, peripheral insulin
resistance is induced. It is considered that obesity results in
impaired glucose tolerance through this mechanism (Matsuzawa Y.:
Diabetes Metab. Rev. 13:3-13, 1997). On the other hand, the onset
mechanism from obesity to hyperlipemia seemingly proceeds as
follows: namely, a mechanism mediated by an increase in the
excessive inflow of the free fatty acids from the visceral fat into
the liver followed by an increase in fat synthesis substrates and
enhancement of expression of lipid synthesis-relating protein
genes; or a mechanism mediated by the promotion of exogenous
cholesterol absorption due to insulin resistance (Kuriyama H. et
al.: Hepatology 27:557-562, 1998).
[0061] When the cross-talk of adipose tissue function and autonomic
nervous activity is taken into consideration from the viewpoint of
factors causative of obesity, the adipose tissue includes the white
adipose tissue serving as a storage of fat energy and the brown
adipose tissue oxidizing and decomposing fat and releasing energy
from the body as heat. The white adipose tissue is widely
distributed all over the body, mainly around visceral tissues and
subcutaneous tissues. It is a specialized organ for storing
excessive energy after food intake and re-supplying it as fatty
acids and glycerol to the whole body. It is also reported that a
decrease in the fat decomposition by the white adipose tissue and a
decrease in the activity of the brown adipose tissue result in
obesity (Nicholls, et al.: Physiol. Rev. 64:1-64, 1984).
[0062] The autonomic nervous activity participate in the control of
the functions of the white adipose tissue and the brown adipose
tissue. Namely, the function of the white adipose tissue is
controlled by the sympathetic nervous and blood hormones, while the
function of the brown adipose tissue is controlled mainly by the
sympathetic nervous (Non-Patent Document 2). When the sympathetic
nervous activity of the white adipose tissue is enhanced, fat
decomposition is promoted. When the sympathetic nervous activity of
the brown adipose tissue is enhanced, fat oxidation and
decomposition are promoted and energy is released from the body as
heat.
[0063] The term "autonomic nervous control effect" as used herein
includes an effect of enhancing sympathetic nervous activity, an
effect of reducing sympathetic nervous activity enhancement, an
effect of reducing sympathetic nervous activity, an effect of
enhancing parasympathetic nervous activity, an effect of reducing
parasympathetic nervous activity enhancement and an effect of
reducing parasympathetic nervous activity. It preferably means an
effect selected from among an effect of enhancing sympathetic
nervous activity, an effect of reducing parasympathetic nervous
activity enhancement and an effect of reducing parasympathetic
nervous activity. More specifically, it means an effect selected
from among an effect of enhancing the sympathetic nervous activity
in the kidney, an effect of enhancing the sympathetic nervous
activity in the adrenal gland, an effect of reducing vagal
(parasympathetic) nervous activity of the stomach, an effect of
enhancing sympathetic nervous activity of the white adipose tissue
and an effect of enhancing sympathetic nervous activity of the
brown adipose tissue.
[0064] Because it contains DDMP, the composition according to the
present invention is usable in treating a disease or condition
relating to autonomic nervous activity; for example, symptoms
frequently occurring due to autonomic nervous disorders such as
shortness of breath, palpitation, shoulder stiffness, headache,
vertigo, anxiety, loss of appetite, lassitude, sleep disorder, etc.
It is particularly useful in treating a disease or condition which
can be treated by enhancing sympathetic nervous activity; for
example, obesity. Also, the composition according to the present
invention reduces the vagal (parasympathetic) nervous activity of
the stomach so as to regulate increased appetite and enhances the
sympathetic nervous activity of the white adipose tissue and the
brown adipose tissue without causing a rise in blood pressure.
Owing to these functions, the composition according to the present
invention promotes fat decomposition in these adipose tissues and
thus prevents fat accumulation, which makes it useful in treating
obesity and/or related complications (for example,
diabetes/impaired glucose tolerance, hyperlipemia, hypertension,
hyperuricemia/gout, coronary artery diseases (heart infarction,
angina pectoris, etc.), brain infarction (brain thrombosis,
transient ischemic attack, etc.), sleep apnea syndrome, fatty
liver, orthopedic diseases (arthritis deformans, lumbar disease,
etc.), menstruation disorder and so on).
[0065] The composition according to the present invention enhances
sympathetic nervous activity of the white adipose tissue so as to
promote fat decomposition. Also, it enhances sympathetic nervous
activity of the brown adipose tissue and promotes oxidation and
decomposition of fat to thereby release energy from the body as
heat, which makes it effective in enhancing energy metabolism. The
enhancement of energy metabolism can be evaluated by, for example,
measuring the oxygen consumption in vivo.
[0066] The expression "to treat a disease or condition" as used
herein means to prevent worsening of the disease or condition, to
relieve the disease or condition, and to prevent the disease or
condition.
[0067] The composition according to the present invention
containing DDMP contains DDMP in a clinically effective amount. The
clinically effective amount means an amount clinically effective in
controlling the autonomic nerve. Based on the results of Examples,
as will be described hereinafter, it is determined that a
sufficient effect can be obtained by administering 0.5 .mu.g/kg or
more of DDMP. Although there is no upper limit of the
administration dose of DDMP, it is generally favorable that the
DDMP dose does not exceed about 50 mg/kg from the viewpoint of the
characteristic smell thereof, and in view of cost.
[0068] To fully achieve the effect, it is desirable that the
composition according to the present invention contains from 0.5
.mu.g/kg to 50 mg/kg per dose (preferably from 1 .mu.g/kg to 10
mg/kg and more preferably from 2 .mu.g/kg to 5 mg/kg) of DDMP. More
specifically, it is desirable that the composition according to the
present invention contains from 30 .mu.g to 3000 mg (preferably
from 60 .mu.g to 600 mg and more preferably from 120 .mu.g to 300
mg) per dose of DDMP in the case of administering to an adult
human. It is possible that the composition according to the present
invention contains from 0.05 .mu.g to 5 mg (preferably from 0.1
.mu.g to 1 mg and more preferably from 0.2 .mu.g to 0.5 mg) of DDMP
per product weight (g). From another viewpoint, it is possible that
the composition according to the present invention contains 25
.mu.g or more (preferably 40 .mu.g or more and more preferably 80
.mu.g or more) of DDMP per product weight (g). In each case, the
upper limit may be adjusted to, for example, 0.5 mg, 1 mg or 5 mg
per product weight (g).
[0069] In the composition according to the present invention, use
can be made of a material containing DDMP, a microorganism capable
of producing DDMP, a culture supernatant of the microorganism, a
culture containing the microorganism, etc. either as such or after
isolation and purification via extraction, fractionation or the
like, though the present invention is not particularly restricted
thereto. It is also possible to use a processed product such as a
concentrate or a dry powder obtained by treating such a material as
described above by using a commonly employed procedure such as
sterilization, vacuum concentration, freeze drying or the like. In
the case of a concentrate which can be hardly powdered by drying,
it may be mixed with an excipient commonly employed in the art such
as dextrin, a high-molecular starch hydrolyzate or a high-molecular
peptide and then powdered by drying. From the viewpoints of
handling properties and storage properties, a powdered composition
is preferred.
[0070] The composition according to the present invention can be
processed into various forms such as foods and drinks (including
foods, drinks, seasonings, alcoholic drinks, functional foods and
so on) and medicinal compositions. For example, it is possible to
provide a food, a drink or a medicine with the use of DDMP, a
microorganism capable of producing DDMP, a culture supernatant of
the microorganism, a processed product thereof and so on.
[0071] Examples of the foods and drinks provided by the present
invention include various ones such as candies, troches, yogurts,
ice creams, puddings, jellies, mizu-yokan (adzuki bean jelly),
alcoholic drinks, coffee drinks, juices, fruit drinks, carbonated
drinks, soft drinks, milk, milk serum drinks, lactic acid drinks
and so on.
[0072] These foods and drinks can be prepared by conventional
methods with the use of various additives if needed. More
specifically, these foods and drinks can be produced by blending of
appropriate materials commonly employed in foods such as glucose,
fructose, sucrose, maltose, sorbitol, stevioside, rubusoside, corn
syrup, lactose, citric acid, tartaric acid, malic acid, succinic
acid, lactic acid, L-ascorbic acid, dl-.alpha.-tocopherol, sodium
erythorbate, glycerol, propylene glycol, a glycerol fatty acid
ester, a polyglycerol fatty acid ester, a sucrose fatty acid ester,
a sorbitan fatty acid ester, a propylene glycol fatty acid ester,
acacia, carrageenan, casein, gelatin, pectin, agar, vitamin Bs,
nicotinamide, calcium panthothenate, amino acids, calcium salts, a
colorant, a perfume, a preserver and so on.
[0073] The medicines provided by the present invention may be in
various dosage forms that are prepared by conventional methods with
the use of various additives if needed. For example, it is possible
to provide medicines for oral administration such as tablets,
capsules, granules, powder, syrups and extracts and medicines for
parenteral administration such as ointments, eye ointments,
lotions, creams, patches, suppositories, eye drops, nasal drops and
injections.
[0074] These medicines may be produced by conventional methods with
the use of various additives. Any additives commonly employed in
the art can be used without specific restriction. Examples thereof
include solid carriers such as starch, lactose, sucrose, mannitol,
carboxymethylcellulose, corn starch and inorganic salts; liquid
carriers such as distilled water, physiological saline, aqueous
glucose solution and alcohols such as ethanol, propylene glycol and
polyethylene glycol; and oily carriers such as various animal or
vegetable oils, white vaseline, paraffin and waxes.
[0075] The composition according to the present invention contains
DDMP as the active ingredient, optionally together with other
active ingredient(s) commonly employed in medicinal compositions,
foods and drinks.
[0076] For example, the composition according to the present
invention may contain one member or a combination of two or more
members selected from among a substance having an autonomic control
effect (a sympathetic agents, a parasympathetic agents, etc.), a
hypotensive substance, an antiobesity substance (a substance
stimulating the appetite regulatory system, a substance acting on
the satiety center, a substance participating in energy metabolism
(for example, a .beta..sub.3 receptor agents, a substance promoting
energy consumption), etc.) that are well known to a person skilled
in the art.
[0077] In the case of the composition according to the present
invention containing a microorganism capable of producing DDMP, a
processed product thereof, a culture supernatant thereof or a
processed product thereof, it may contain, in particular, the
above-described substances with microbial origin (a substance
having an autonomic control effect, a hypotensive substance, a
substance promoting energy metabolism, etc.).
[0078] In the medicinal composition, food or drink according to the
present invention, indication may be made of a specific purpose of
use (for example, controlling the autonomic nerve, enhancing the
sympathetic nervous activity, regulating the parasympathetic
nervous activity, preventing obesity, treating obesity, treating a
complication of obesity, promoting energy metabolism, promoting fat
burning or maintaining health) and/or a specific way of use (for
example, dose, dosing frequency and dosing method).
EFFECTS OF THE INVENTION
[0079] As discussed above in detail, the present invention provides
a medicinal composition, a food or a drink for controlling
autonomic nervous activity per se (in particular, enhancing the
sympathetic nervous activity) that are the fundamental factors
causative of a disease or condition relating to the autonomic
nervous activity. In particular, it provides a medicinal
composition, a food or a drink having an effect of enhancing the
sympathetic nervous activity of the white adipose tissue and the
brown adipose tissue without causing a rise in blood pressure.
Further, it provides a medicinal composition, a food or a drink
having an effect of promoting energy metabolism. Thus, it provides
a medicinal composition, a food or a drink which is efficacious and
highly safe in treating obesity and/or related complications with
little side effect (no rise in blood pressure) and can be
conveniently produced.
BRIEF DESCRIPTION OF THE DRAWINGS
[0080] FIG. 1 presents an MS spectrum of
2,3-dihydro-3,5-dihydroxy-6-methyl-4H-pyran-4-one obtained in
Example 1.
[0081] FIG. 2 presents NMR spectra ((a) showing a 13C-NMR spectrum,
while (b) showing a 1H-NMR spectrum) of
2,3-dihydro-3,5-dihydroxy-6-methyl-4H-pyran-4-one obtained in
Example 1.
[0082] FIG. 3 is a graph which shows the effect of enhancing the
rat kidney sympathetic nervous activity of
2,3-dihydro-3,5-dihydroxy-6-methyl-4H-pyran-4-one obtained in
Example 1.
[0083] FIG. 4 is a graph which shows that
2,3-dihydro-3,5-dihydroxy-6-methyl-4H-pyran-4-one obtained in
Example 1 causes no rise in blood pressure.
[0084] FIG. 5 is a graph which shows the effect of reducing rat
stomach vagal (parasympathetic) nervous activity of
2,3-dihydro-3,5-dihydroxy-6-methyl-4H-pyran-4-one obtained in
Example 1.
[0085] FIG. 6 is a graph which shows the effect of enhancing
sympathetic nervous activity of the rat interscapular brown adipose
tissue of 2,3-dihydro-3,5-dihydroxy-6-methyl-4H-pyran-4-one
obtained in Example 1.
[0086] FIG. 7 is a graph which shows the effect of enhancing the
rat white adipose tissue sympathetic nervous activity of
2,3-dihydro-3,5-dihydroxy-6-methyl-4H-pyran-4-one obtained in
Example 1.
[0087] FIG. 8 is a graph which shows the effect of promoting rat
energy metabolism of
2,3-dihydro-3,5-dihydroxy-6-methyl-4H-pyran-4-one obtained in
Example 1.
EXAMPLES
[0088] Now, the present invention will be described in greater
detail by reference to the following Examples, though the present
invention is not restricted thereto.
[0089] In the present Examples,
2,3-dihydro-3,5-dihydroxy-6-methyl-4H-pyran-4-one was prepared by
using a culture supernatant of Lactobacillus plantarum SAM 2446
strain (FERM ABP-10438) as the starting material and conducting a
fractionation treatment. Then, the effects of
2,3-dihydro-3,5-dihydroxy-6-methyl-4H-pyran-4-one on rat autonomic
nervous activity were examined.
[0090] In the present Examples, moreover,
2,3-dihydro-3,5-dihydroxy-6-methyl-4H-pyran-4-one was prepared by
using glucose, piperidine and thioglycolic acid. Then, the effects
of 2,3-dihydro-3,5-dihydroxy-6-methyl-4H-pyran-4-one on rat energy
metabolism was examined.
[0091] In the present Examples, furthermore, medicines, foods and
drinks were produced using DDMP.
Example 1
Preparation of 2,3-dihydro-3,5-dihydroxy-6-methyl-4H-pyran-4-one
(Purification from Lactic Acid Bacterium Culture Supernatant)
[0092] As the lactic acid bacterium culture supernatant, use was
made of a freeze-dried culture supernatant obtained by culturing
Lactobacillus plantarum SAM 2446 strain (FERM ABP-10438) in MRS
medium. A sterilized MRS medium was inoculated with Lactobacillus
plantarum SAM 2446 strain (FERM ABP-10438) which was then
statically cultured therein at 37.degree. C. for 1 day. Next, the
cells were removed by centrifugation to give a culture supernatant.
By use of the culture supernatant thus obtained, a freeze dried
product was prepared by a conventional method and employed as the
starting material. First, the lactic acid bacterium culture
supernatant was ultra-filtered using Ultrafiltration Membranes YM10
(manufactured by Millipore) to give a low molecular fraction
passing through the membranes. Next, the obtained fraction was
chromatographically fractionated through Develosil C30 UG-5 Column
(manufactured by NOMURA CHEMICAL Co., Ltd.) with the use of the
effect on the kidney sympathetic nervous activity observed in
transduodenal administration as an indication, thereby giving
2,3-dihydro-3,5-dihydroxy-6-methyl-4H-pyran-4-one. FIGS. 1 and 2
present MS and NMR data of this compound. In this method, the yield
of 2,3-dihydro-3,5-dihydroxy-6-methyl-4H-pyran-4-one was 1.78 mg
per 100 ml of the lactic acid bacterium culture supernatant.
Example 2
Evaluation of the effects of
2,3-dihydro-3,5-dihydroxy-6-methyl-4H-pyran-4-one on Rat Kidney
Sympathetic Nervous Activity, Blood Pressure and Stomach Vagal
(Parasympathetic) Nervous Activity
[0093] In these experiments, use was made of male Mister rats
weighing about 300 g which had been fed in a thermostatic chamber
(24.degree. C.) with a light-dark cycle of 12-hour intervals (under
illumination from 8:00 to 20:00) for 1 week or longer. A single rat
was employed for each nervous experiment. The animals were fed with
a feed (Oriental Yeast, MF) and water ad libitum. The autonomic
nervous activity were examined by fasting the rats for 3 hours on
the day of the experiment and then subjecting to an abdominal
surgery under urethane-anesthesia at the intermediate point of the
light period. Then, the effects of the transduodenal administration
of 2,3-dihydro-3,5-dihydroxy-6-methyl-4H-pyran-4-one obtained in
Example 1 on the kidney sympathetic nervous activity, stomach vagal
(parasympathetic) nervous activity and blood pressure were measured
in accordance with the method reported by T. Yamano et al.,
Neurosci. Lett. 313:78-82 (2001), A. Niijima et al., Autonom.
Neurosci.: Basic & Clinical 97:99-102 (2002) and M. Tanida et
al., Am. J. Physiol: Regulatory, Integrative and Comparative
Physiology 288:R447--R455 (2005). Namely, the sympathetic nerve
controlling the kidney and the parasympathetic nerve controlling
the stomach were picked up using a silver electrode. A cannula was
inserted into the left femoral artery and then the blood pressure
was measured by use of a transducer. From the initiation of the
surgery to the completion of the measurement, a tube was inserted
into the respiratory tract for the management of the airway. The
body temperature (rat rectal temperature) was maintained at
35.+-.0.5.degree. C. with the use of a heater. The transduodenal
administration was conducted by dissolving
2,3-dihydro-3,5-dihydroxy-6-methyl-4H-pyran-4-one obtained in
Example 1 in physiological saline to give a concentration of 10
.mu.g/2 ml, and administering 2 ml/animal of the obtained solution
at a rate of 1 ml/min through a polyethylene tube inserted into the
duodenum. Electrical nervous activity thus obtained were amplified
with a condenser type differential amplifier, monitored with an
oscilloscope and recorded on a magnetic tape. To separate the raw
data from the background noise, all of the nervous activity were
converted into standard pulses by use of a slicer and a window
discriminator before analysis. Discharge frequencies were indicated
on a pen recorder by use of a rate meter at a reset time of 5
seconds. The data was analyzed based on the average discharge
frequency per 5 seconds (pulse/5 sec) and expressed as a percentage
by referring to the level before the administration as to 100%. A
group with the administration of physiological saline was employed
as a control.
[0094] As a result, the control group showed no effect on all of
the autonomic nervous activity and blood pressure. In the
administration group, on the other hand, the kidney sympathetic
nervous activity was enhanced (FIG. 3) while the blood pressure was
not elevated (FIG. 4) and the stomach vagal (parasympathetic)
nervous activity was reduced (FIG. 5).
Example 3
Evaluation of the Effects of
2,3-dihydro-3,5-dihydroxy-6-methyl-4H-pyran-4-one on Rat
Interscapular Brown Adipose Tissue Sympathetic Nervous Activity and
Epididymal White Adipose Tissue Sympathetic Nervous Activity
[0095] The procedure of Example 2 was followed and thus the effects
of the transduodenal administration (10 .mu.g) of
2,3-dihydro-3,5-dihydroxy-6-methyl-4H-pyran-4-one obtained in
Example 1 on the rat epididymal white adipose tissue sympathetic
nervous activity and the interscapular brown adipose tissue
sympathetic nervous activity were examined in accordance with the
methods reported in the documents cited in Example 2. A single rat
was employed for each nervous experiment.
[0096] As a result, both of the interscapular brown adipose tissue
sympathetic nervous activity (FIG. 6) and the epididymal white
adipose tissue sympathetic nervous activity (FIG. 7) were
enhanced.
Example 4
Preparation of 2,3-dihydro-3,5-dihydroxy-6-methyl-4H-pyran-4-one
(Synthesis with the Use of Glucose, Piperidine and Thioglycolic
Acid)
[0097] 2,3-Dihydro-3,5-dihydroxy-6-methyl-4H-pyran-4-one was
prepared by use of glucose, piperidine and thioglycolic acid in
accordance with Isabelle Cutzach et al.: J. Agric. Food Chem.
47:1663-67, 1999 (modified). Next the synthesis method reported in
this document will be briefly described.
[0098] To a suspension of D-glucose (600 g, 3.33 mol) and
triethylamine (192 mL, 3.33 mol), piperidine (329 mL, 3.33 mol) and
acetic acid (192 mL, 3.33 mol) were added under stirring at room
temperature. The obtained mixture was stirred under heating at
70.degree. C. for 0.1 hour. After cooling to room temperature by
standing, the mixture was allowed to stand in a refrigerator
overnight. The solid thus precipitated was washed with 4 L of a
solvent mixture of ethanol-acetone (1:1) and collected by
filtration. Then, it was heated to 35.degree. C. and dried under
reduced pressure to give a piperidine derivative.
[0099] A solution of the obtained piperidine derivative (176 g,
0.711 mol) in ethanol (1.6 L) was stirred under heating at
75.degree. C. A solution of thioglycolic acid (49.6 ml, 0.713 mol)
in ethanol (135 mL) was dropped thereinto over 40 minutes and the
mixture was heated to 75.degree. C. and stirred for additional 20
hours. After completion of the reaction, the mixture was
concentrated under reduced pressure to give crude
2,3-dihydro-3,5-dihydroxy-6-methyl-4H-pyran-4-one. The crude
2,3-dihydro-3,5-dihydroxy-6-methyl-4H-pyran-4-one thus obtained was
subjected to column chromatography successively with Daiso SP
120-40/60-ODS-B (manufactured by DAISO) and CHP20P (manufactured by
MITSUBISHI CHEMICAL Co.). Thus a purified
2,3-dihydro-3,5-dihydroxy-6-methyl-4H-pyran-4-one preparation was
obtained.
[0100] In the above synthesis method, the purified
2,3-dihydro-3,5-dihydroxy-6-methyl-4H-pyran-4-one preparation was
obtained at a yield of 6.93 g of the purified
2,3-dihydro-3,5-dihydroxy-6-methyl-4H-pyran-4-one preparation per
600 g of glucose.
Example 5
Effect of Promoting Energy Metabolism of
2,3-dihydro-3,5-dihydroxy-6-methyl-4H-pyran-4-one
[0101] Male SD rats aged 7 weeks were preliminarily fed with a
solid feed (CE-2: NIPPON CREA) and water ad libitum for 1 week for
the habituation to the light-dark cycle of 12-hour intervals. The
thus habituated animals were fasted for 16 hours before the
administration of the samples and then divided into two groups.
Then, 40 .mu.g/8 ml/kg body weight of DDMP and 8 ml/kg body weight
of distilled water for injection were forcibly orally administered
into the stomach to the respective groups. Within 60 minutes
following the administration, oxygen consumption was measured with
a metabolic meter for small animals (MK-5000RQ2: manufactured by
MUROMACHI KIKAI Co., Ltd.). A combination of a DDMP-administered
rat with a water-administered rat was referred to as a single
experimental lot and the averages of five combinations (i.e., a
DDMP-administered group having 5 rats and a water-administered
group having 5 rats) were calculated.
[0102] As a result, the DDMP-administered group showed an increase
in oxygen consumption compared with the water-administered group.
This difference was observed from 15 to 33 minutes after the
administration (FIG. 8). Based on these results, it was confirmed
that DDMP has an effect of promoting energy metabolism.
Production Example 1
DDMP-Containing Medicine
Tablet:
[0103] A DDMP-containing medicine (tablets) was produced by the
following method.
[0104] 2 g of DDMP obtained in Example 4, 296.7 g of lactose and
1.3 g of magnesium stearate were mixed together and tabletted with
a single-shot tabletting machine to give tablets, each being 10 mm
in diameter and 300 mg in weight.
[0105] Granules:
[0106] 2 g of DDMP obtained in Example 4 was added to 296.7 g of
lactose and 1.3 g of magnesium stearate and the mixture was
compressed, powdered, dressed and sieved to give granules of 20-50
mesh.
Production Example 2
DDMP-Containing Food and Drink
[0107] Various DDMP-containing foods and drinks of the following
compositions were produced by conventional methods.
TABLE-US-00002 (Component) (Part by weight) Ice cream: Fresh cream
(fat: 45%) 33.8 Skim milk powder 11.0 Granulated sugar 14.8
Sugar-containing yolk 0.3 Vanilla essence 0.1 Water 39.998 DDMP
0.002 Total 100.00 Juice: Unshu orange juice concentrate (frozen)
5.0 High-fructose corn syrup 11.0 Citric acid 0.2 L-Ascorbic acid
0.02 DDMP 0.002 Perfume 0.2 Colorant 0.1 Water 83.478 Total 100.00
Lactic acid drink: Fermented milk (solid milk content: 21%) 14.76
High-fructose corn syrup 13.31 Pectin 0.5 Citric acid 0.08 Perfume
0.15 Water 71.198 DDMP 0.002 Total 100.00 Yogurt: Fresh milk (fat:
3.4%) 80.0 Fresh cream (fat: 50%) 8.0 Skim milk powder 1.5 Water
7.498 Starter 3.0 DDMP 0.002 Total 100.00 Coffee drink: Granulated
sugar 8.0 Skim milk powder 5.0 Caramel 0.2 Coffee extract 2.0
Perfume 0.1 Polyglycerol fatty acid ester 0.05 Sodium chloride 0.05
Water 84.598 DDMP 0.002 Total 100.00 Alcoholic drink: 50 vol. %
ethanol 32.0 Sugar 8.4 Fruit juice 2.4 DDMP 0.002 Purified water
57.198 Total 100.00
Discussion
[0108] Although 2,3-dihydro-3,5-dihydroxy-6-methyl-4H-pyran-4-one
obtained from the lactic acid culture supernatant in Example 1
enhanced the kidney sympathetic nervous activity, it caused no rise
in blood pressure. Based on these results, it is expected to have a
higher safety level relative to hypertension, which brings about a
problem as a complication of obesity, than commonly employed
compositions having an effect of enhancing sympathetic nervous
activity. Since it showed an effect of reducing vagal
(parasympathetic) nervous activity of the stomach, it is expected
to have an effect of intestinal canal-inhibitively regulating
appetite. Moreover, it showed effects of enhancing sympathetic
nervous activity of the interscapular brown fat adipose and the
epididymal white adipose tissue. Thus, it is expected to have an
antiobesity effect through the enhancement of fat burning. The
procedures employed in the above Examples 2 and 3 are also
advantageous in that the autonomic nervous control functions of
different substances can be successively evaluated so long as the
test animal survives.
[0109] It has been further clarified that
2,3-dihydro-3,5-dihydroxy-6-methyl-4H-pyran-4-one enhances energy
metabolism. These results indicate that use of
2,3-dihydro-3,5-dihydroxy-6-methyl-4H-pyran-4-one enables the
provision of a medicinal composition, a food or a drink which is
efficacious against obesity and/or related complications.
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