JP2002265385A - Qol improving preparation - Google Patents

Abstract

PROBLEM TO BE SOLVED: To provide a QOL(quality of life) improving preparation preferably in the form of a food/drink intended to improve QOL by activating mainly immunological function, and to provide a new application of an immunoactivating substance characterized by using it for improving QOL. SOLUTION: This QOL improving preparation is characterized by comprising an immunoactivating substance. The other objective application of such an immunoactivating substance for improving QOL is provided.

Description

DETAILED DESCRIPTION OF THE INVENTION

[0001]

The present invention relates to a QOL improver which is preferably used especially as a food or drink, and to the use of an immunostimulating substance for improving the QOL. Here, QOL stands for quality of life.

[0002]

2. Description of the Related Art In recent years, attention has been focused on improving the quality of life from a medical point of view. In this case, the quality of life is the quality of life of a person having a disease, and the objective result (such as various test values, etc., ), Plus the mental, psychological,
It is an index of satisfaction obtained by comprehensively judging the status of social activities. As a QOL evaluation method, a method of listening to a subjective health degree through a questionnaire survey and quantifying the result is used.

[0003] By such a questionnaire survey, the QOL of a healthy person can be measured, but not all people show a high value. The first thing that can be considered from these results is that there is a large range in the degree of health. For example, unbalanced diet, high-calorie high-fat diet, eating habits such as irregular meal time, lack of exercise, stress, and various unfavorable lifestyles such as smoking and drinking affect human health, QOL may be reduced even without illness. In addition, the number of people suffering from chronic diseases such as allergies is increasing with the deterioration of the environment, and the discomfort caused by such chronic diseases is one of the causes of lowering QOL. Furthermore, even if one takes great care of the lifestyle, the decline of biological functions with aging is unavoidable, which may cause illness, if not illness, and lower the QOL.

[0004]

SUMMARY OF THE INVENTION An object of the present invention is to provide a QOL improver intended to improve QOL by activating an immune function, preferably in the form of food or drink. An object of the present invention is to provide a new use of an immunostimulatory substance characterized by using an immunostimulatory substance for improving QOL.

[0005]

Means for Solving the Problems The present inventors have conducted intensive studies on the improvement of QOL of a so-called sick person as well as the improvement of the QOL of a healthy person. (For example, bacteria, viruses or molds), and is caused (even slightly) by a decrease in immune function associated with the above-mentioned undesirable lifestyle, aging or allergic disease. We found that opportunistic infections are a major cause of daily discomfort. As a result of further examination of such immunological findings, it was found that if a substance having an immunostimulatory effect was added to foods and drinks taken daily to maintain and improve the immune function, QOL
Has been obtained, and it has been found that health can be maintained and promoted. The present inventors have further studied and completed the present invention.

That is, the present invention relates to (1) a QOL improver characterized by containing an immunostimulating substance, (2) NK
A QOL enhancer containing an immunostimulatory substance that increases the cytotoxic activity of cells by 10% or more and / or an immunostimulatory substance that causes lymphocytes to proliferate by 10% or more in a lymphocyte co-stimulation test; The activator is 3-O
The QOL improver according to the above (1) or (2), which is a saccharide containing -α-D-glucopyranosyl-D-glucose as a constituent unit, and (4) the saccharide comprises nigerose, nigerosyl glucose, and nigerosyl maltose. The QOL improver according to the above (3), which is a mixture of at least one kind of nigerooligosaccharide selected from the group, and (5) the Q according to the above (1) to (4), which is a food or drink.
The present invention relates to an OL enhancer, (6) use of an immunostimulating substance for improving QOL, and (7) use of the QOL improver described in (1) to (4) for maintenance / promotion of health.

[0007]

BEST MODE FOR CARRYING OUT THE INVENTION The immunostimulating substance used in the present invention is not particularly limited. For example, activated hemicellulose obtained by enzymatically treating a plant fiber contained in a cell wall of a mushroom mycelium, a mycelium component of a sarcoma moss PSK, SPG which is a mycelial component of Suehirotake, lentinan which is a mycelial component of Shiitake, or mycelial components such as Agaricus, Reishi, Ningyotake, Kawariharatake or Maitake, and OK which is a bacterial component of Streptococcus
-432, lactic acid bacteria or a processed product thereof, 3-O-α-D
And saccharides containing -glucopyranosyl-D-glucose as a structural unit. Among them, lactic acid bacteria or processed products thereof, or saccharides containing 3-O-α-D-glucopyranosyl-D-glucose as a constituent unit are preferably used as the immunostimulating substance according to the present invention.

Here, the saccharide containing 3-O-α-D-glucopyranosyl-D-glucose as a constituent unit is not particularly limited, and those known per se may be used.
Specifically, for example, at least one or more α-1,3
Oligosaccharides containing a glucosidic bond and having a degree of glucose polymerization of about 2 or more can be mentioned.
Oligosaccharides having a degree of polymerization of about 0, more preferably nigerooligosaccharides, which are oligosaccharides having a degree of polymerization of about 2 to 7, are preferably used in the present invention. Such nigerooligosaccharides include α
In addition to oligosaccharides consisting only of -1,3 glucoside bonds,
α-1,3 glucoside bond and other bond (for example, α
-1,1, α-1,2, α-1,4, α-1,6 glucosidic bond). Among them, in the present invention, nigerose represented by the following formula,
More preferably, nigerosyl glucose or nigerosyl maltose is used. These may be used alone, or two or more kinds may be used in combination or as a mixture.

[0009]

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[0010]

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[0011]

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The saccharide containing 3-O-α-D-glucopyranosyl-D-glucose as a structural unit used as an immunostimulating substance in the present invention can be easily produced according to a method known per se. Specifically, for example, a nigerooligosaccharide preferable as the saccharide used in the present invention can be prepared by the following known method. For example, M. Stacey and JMWebber: Meth
ods in Carbohydrate Chemistry, I, 339-341, AcademicPr
ess (1962) proposes a method for producing a nigerooligosaccharide by hydrolyzing a polysaccharide produced by a microorganism, such as nigeran or ercinan, as a substrate using an enzyme or an acid. Also known is a method for preparing nigerose using a known α-glucosidase transglycosylation / condensation reaction (Kanaya Kenichi et al., Journal of the Japanese Society of Agricultural Chemistry, 53,
385-390 (1979), H. Fujimoto et al., Agric. Biol. Ch.
em., 52, 1345-1351 (1988)). Further, Japanese Unexamined Patent Application Publication No.
Japanese Patent No. 22958 discloses a method for producing nigerose by allowing a cyclodextrin-forming enzyme to act on a starch hydrolyzate. In addition, one or two or more of glycosyltransferases that provide an α-1,3 glucoside bond to a substrate containing an α-1,4 glucoside-linked polysaccharide or oligosaccharide, specifically Acremoni
um belonging to the genus um and producing a glycosyltransferase that produces an α-1,3 bond, such as Acremonium sp. S4G13 (FERM BP-437).
A method for producing a nigerooligosaccharide by reacting a glycosyltransferase prepared by culturing 3) according to a conventional method has also been disclosed (JP-A-7-59559). The nigerooligosaccharide used in the present invention may be prepared by any method, and is not limited to the above method. However, the most economical method known to date is considered to be the one described in the above-mentioned Japanese Patent Application Laid-Open No. 7-59.
This is a method using a glycosyltransferase (nigero-oligosaccharide-forming enzyme) described in Japanese Patent Application Laid-Open No. 559 or JP-A-2000-189101. In the present invention, it is preferable to use a nigerooligosaccharide prepared according to this method.

A preferred embodiment of the immunostimulating substance used in the present invention is a substance having a lymphocyte costimulatory effect. More specifically, in a lymphocyte (T lymphocyte and / or B lymphocyte) co-stimulation test, lymphocytes proliferate by about 10% or more, preferably about 15% or more, more preferably about 15 to 30%. The substance that causes it is preferred.

The lymphocyte costimulatory effect can be measured according to a method known per se. Specific examples of the measurement method include a lymphocyte co-stimulation test for the mouse spleen lymphocyte antigen-specific proliferation reaction described below. That is, the spleen is aseptically removed from the mouse, crushed in an RPMI 1640 medium, and passed through a 200-mesh screen to obtain a spleen cell suspension. After measuring the number of cells in the spleen cell suspension using an automatic hemacytometer, adjust the cell number to a concentration of 1 × 10 ⁷ / ml with RPMI 1640 medium (the resulting solution is referred to as “spleen cell suspension preparation”). ), And inoculate 50 μl per well into a 96-well tissue culture plate. A solution prepared by dissolving anti-B lymphocyte antigen receptor immunoglobulin M (anti-IgM) at a concentration of 200 μg / ml in RPMI 1640 medium was added in an amount of 50 μl per well.
Each of the spleen cell suspensions sown above was added to a B lymphocyte stimulation group.

[0015] The T lymphocyte stimulation group contains anti-T
Lymphocyte antigen receptor monoclonal antibody (anti-CD3)
A solution dissolved in borate buffer at a concentration of 10 μg / ml was added to a 96-well tissue culture plate in an amount of 100 μl per well, and allowed to stand at 37 ° C. for 3 hours to allow anti-CD3 to adhere to each well. RPM
After washing with I 1640 medium, add 50 μl of RPMI 1640 medium per well.
The spleen cell suspension preparation is seeded at 50 μl per well in the wells added.

To each of the two groups, a solution prepared by dissolving the immunostimulatory substance of the present invention at a concentration of 4 mg / ml in RPMI 1640 medium was added in an amount of 100 μl per well, and this was used as a test group. In addition, RPMI 1640 medium was added in an amount of 100 μl per well, and this was used as a comparative group. 37 ° C for these test and comparative groups
Culture for 2 days in a 5% CO 2 incubator. End culture
3 hours ago 3- (4,5-dimethyl-2-thiazolyl) -bromide
A solution prepared by dissolving 2,5-diphenyl-2H tetrazolium at a concentration of 5 mg / ml in RPMI 1640 medium was added at 10 μl / well, and at the end of the culture, a 20% by weight sodium dodecyl sulfate solution was added at 50 μl / well. After standing at ℃ for 1 day, the proliferation reaction of spleen cells is determined by measuring the absorbance of the culture solution at 550 nm using a microplate reader.

Another preferred embodiment of the immunostimulant used in the present invention is a substance having a natural killer cell (hereinafter referred to as "NK cell") activating effect. More specifically, an immunostimulatory substance capable of increasing the cytotoxic activity of NK cells by about 10% or more, preferably about 15 to 30%, more preferably about 20 to 30% is preferable.

The cytotoxic activity of such NK cells can be easily measured by those skilled in the art, for example, by measuring a natural killer activity using a fluorescent dye method. Specifically, the following measuring method can be used.
The liver is removed from the mouse, crushed in Hanks Balanced Salt Solution, and passed through a # 225 mesh. After centrifugation, the cells are resuspended in a 36% by weight Percoll-Hanks balanced salt solution and centrifuged to precipitate the liver mononuclear cell fraction. 0 to this fraction
After hemolysis with an 83% by weight ammonium chloride buffer, the suspension is suspended in RPMI 1640 medium to obtain a suspension of liver mononuclear cells. After measuring the cell count with an automatic hemocytometer, 2.0 x 1
0 ^6, 5.0 × 10 ^5, the concentration of 12.5 × 10 ⁴ / ml was prepared in RPMI 1640 medium, 40 × natural killer activity measurement, 10 × and
Use 2.5 × effector cell solution. For each well of a 96-well tissue culture V-bottom plate, 25 μl of each effector cell solution or 25 μl of 0.4% by weight of Triton X-100 dissolved in RPMI 1640 medium (for maximum fluorescence measurement) or RP
Seed 25 μl of MI 1640 medium (for minimum fluorescence measurement).
To this, the immunostimulating substance according to the present invention was mixed with RP at a concentration of 4 μg / ml.
Add 25 μl per well of the solution dissolved in MI 1640 medium,
This is referred to as a test group. Further, only 25 μl of RPMI 1640 medium was added per well, and this was used as a comparative group.

YAC-1 cells were used as target cells, and 1.0 × 10 ⁶ /
YAC-1 cells adjusted to a concentration of ml are labeled with 15 μM calcein AM dye for 30 minutes, washed, and suspended in RPMI 1640 medium to adjust the cell number to a concentration of 2.5 × 10 ⁴ / ml. This target cell was added to each of the test group and the comparative group in an amount of 5 per well.
Add 0 μl each to start the natural killer reaction. The reaction was incubated in a 5% CO2 incubator at 37 ° C for 3 hours, 50 μl of the supernatant was collected, and the amount of eluted calcein AM dye was detected by fluorescence measurement at an excitation wavelength of 490 nm and a fluorescence wavelength of 530 nm. I do. The cytotoxic activity is determined from the following equation. [Equation 1] Cytotoxic activity (%) = [(fluorescence value when effector cells are added−minimum fluorescence value) / (maximum fluorescence value−minimum fluorescence value)] ×
100

Originally, QOL was a term having a broad concept that encompassed living conditions such as economic conditions, occupations, and residences.
The QOL in the present invention refers to “Health related QOL” used in connection with health or disease.

The QOL improver of the present invention can be used to
In addition to improving QOL, it has an effect of improving QOL of, for example, a sick person. Specifically, the improvement and improvement of the QOL are determined by the fact that when the QOL improver of the present invention is administered or ingested, the numerical value obtained by quantifying the questionnaire results of the subjective health level substantially increases. can do.

More specifically, it can be determined by the following method. Headache / heavy weight, dizziness / dizziness, stiff shoulders, palpitations,
Facial flushing, numbness, swelling, pruritus, shortness of breath, constipation, pain, swelling, stiffness in joints, tired immediately after walking a short distance, throat dry, appetite, food is delicious Do you remain tired when you wake up in the morning, are you comfortable with falling asleep, do you wake up at night with urination (piss), wake up soon after you sleep, do you often have bad dreams, do you feel good? 24 questions about whether you are annoyed, somehow anxious, or depressed,
A questionnaire survey prepared with three-step answers (frequent, sometimes, not so often) was asked by each person who administered or ingested the QOL improver of the present invention at the start and end of the tasting. From the highest QOL
Score 2 points, 1 point, and sum the scores of the above 24 question items to obtain a total QOL score. If the total QOL score increases by 2 points or more before and after ingestion or administration, it is considered that the QOL has improved. to decide.

Here, "3 points in descending order of QOL, 2 points
"Score to one point" means, for example, in the question item of "headache / heavy weight", 3
Two points are given for points, "sometimes", and one point for "frequently". On the other hand, the question item "Do you have an appetite?"
The answer "sometimes" is given 2 points, and the answer "not so much" is given 1 point.

The QOL improver according to the present invention can be used not only as a medicine but also as a food or drink. More specifically, for example, seasonings, processed meat, processed fishery products,
It is also possible to provide the QOL improver according to the present invention in the form of processed agricultural products, staples, seasoned foods, cooked foods, desserts, dairy products, confections or snack confections. In particular, the QOL improver according to the present invention is preferably used as a food or drink.

The QOL improver according to the present invention is characterized in that it contains an immunostimulating substance, and further contains other known foods or food components, pharmaceutical carriers or excipients, for example. May be. Such other components are not particularly limited, and the desired QOL
Those skilled in the art can appropriately select them according to the specific use of the enhancer. Specifically, in the case of pharmaceuticals, excipients, disintegrants, binders, surfactants, emulsifiers, plasticizers, lubricants and saccharides , PH
In the case of food, various nutrients, vitamins, flavors, coloring agents, antioxidants, flavor substances such as cheese and chocolate, and synthetic sweeteners are exemplified.

The dosage form of the QOL enhancer according to the present invention may be oral, such as syrup, powder, granule, pill, tablet, hard capsule or soft capsule, depending on the method of ingestion or administration and the route of ingestion or administration. Preparations, suppositories, injections and the like, but are preferably used as oral preparations. In the case of taking the form of a pill or a tablet, it may be an orally disintegrating tablet. Further, when the QOL improver of the present invention is used as a food, granules,
It can be provided in the form of tablets, gums, candy, jelly, beverages, etc., but the form is not limited to the above.

[0027] The intake or dosage of the QOL improver of the present invention varies depending on the kind of the immunostimulating substance, the sex, age, and health condition of the person to be ingested or administered. What is necessary is just to select suitably from the range which the intake or dosage of a substance becomes an effective amount. The number of times of ingestion or administration may be once a day, or may be plural times.

[0028]

EXAMPLES The present invention will be described in detail below with reference to examples, but the present invention is not limited to these examples. In the present example, elderly people 65 years or older were allowed to eat and drink nigerooligosaccharides, and the QOL improvement effect of the nigerooligosaccharides was verified by examining the QOL before and after sampling. The study group included 25 healthy elderly people aged 65 years or older who obtained written informed consent from all participants prior to the start of the study. Content 30.8% by weight) [sold by Takeda Food Industry Co., Ltd.], and the control group was Hymaltose Syrup MC-45 (Nippon Shokuhin Kako Co., Ltd.).
A single-blind, placebo-controlled, comparative study was performed using
The control group (10 g control product / day ingested, 12 people) and the test group (10 g test product / day ingested, 13 people) were randomly assigned, the tasting period was 4 weeks, and the time of ingestion was arbitrary.

At the start and end of the tasting, 24 questions prepared for each taster in questionnaire questionnaire in three stages (headache / head weight, dizziness / dizziness, stiff shoulders, palpitations, hot flush, numbness, swelling) , Itching, shortness of breath, constipation,
Joint pain, swelling, stiffness, tiredness when walking a short distance, dry throat, appetite,
Are the foods good, are you tired when you wake up in the morning, are you good at falling asleep, get up at night urinating (piss), wake up soon after you sleep, do you often have bad dreams, Are you feeling refreshed or frustrated?
Do you feel anxious or feel depressed?) And evaluate your QOL. Answers were scored as 3 points, 2 points, and 1 point from the highest QOL, and the scores of the above 24 question items were totaled to obtain a total QOL score. Subtract the total QOL score before the tasting from the total QOL score after the tasting of each taster, and if the difference is 2 or more, the QO
If L is improved and the difference is 0 or 1 point, QO
When L is unchanged, if the difference is less than 0, it is determined that QOL has decreased. Intra-group comparison of total QOL scores was performed by comparing the total QOL scores before and after the tasting using the Wilcoxon signed rank sum test, and comparison between groups of QOLs was defined as three points of increase, invariance, and decrease in the total QOL score after tasting. The data were converted to a two-point, one-point ordinal scale and compared between the control group and the test group using the Mann-Whitney U test.

Table 1 shows the total score of QOL before and after the tasting, and Table 2 shows the number of persons whose QOL improved, remained unchanged, and decreased after the tasting. As is evident from Tables 1 and 2, the QOL of the nigero-oligosaccharide-containing syrup taster was improved, and the effect of eating and drinking the nigerooligosaccharide on the QOL was verified.

[0031]

[Table 1]

[0032]

[Table 2]

[0033]

As the life has been enriched and diversified in recent years, it has been inevitable to some extent that lifestyle habits have deteriorated or become irregular. In addition, allergic diseases caused by environmental pollution are also increasing. Furthermore, with the advent of an aging society, the number of people who complain of physical disorders due to the deterioration of physiological functions due to aging is increasing year by year, and is expected to increase further in the future. In view of such a social situation, the QOL improver according to the present invention is very useful not only for a so-called sick person but also for a healthy person. Further, the present invention can provide a new use of the immunostimulatory substance, in which the immunostimulatory substance is used for improving QOL.

──────────────────────────────────────────────────続 き Continued on the front page (51) Int.Cl. ⁷ Identification symbol FI Theme court ゛ (Reference) // C07H 3/04 C07H 3/04 3/06 3/06 (72) Inventor Norio Yamamoto Kobe, Hyogo Prefecture 4B018 LB01 LB05 LB06 LB08 LB09 LE02 MD31 ME07 4C057 BB03 BB04 4C084 AA17 NA14 ZA022 ZB092 4C086 AA01 AA02 EA01 MA01 MA02 MA03 MA04 NA14 ZAB ZB

Claims (7)

[Claims] 1. A QOL improver comprising an immunostimulating substance. 2. An improvement in QOL comprising an immunostimulatory substance that increases the cytotoxic activity of NK cells by 10% or more and / or an immunostimulatory substance that causes lymphocytes to proliferate by 10% or more in a lymphocyte co-stimulation test. Agent. 3. The QOL improver according to claim 1, wherein the immunostimulatory substance is a saccharide containing 3-O-α-D-glucopyranosyl-D-glucose as a constituent unit. 4. The QOL improver according to claim 3, wherein the saccharide is a mixture of at least one kind of nigerooligosaccharide selected from the group consisting of nigerose, nigerosyl glucose and nigerosyl maltose. 5. The QO according to claim 1, which is a food or drink.
L improver. 6. Use of an immunostimulating substance for improving QOL. 7. A method for maintaining and promoting health.
Use of the QOL improver described in 1.