JP2008024620A - Adiponectin production inhibitor, skin care preparation, bath preparation, food and drink, and fodder - Google Patents

Abstract

<P>PROBLEM TO BE SOLVED: To provide an adiponectin production inhibitor useful for prevention, mitigation and amelioration of various kinds of disorders, diseases, symptoms and the like caused by adiponectin. <P>SOLUTION: The adiponectin production inhibitor contains a crude drug of Rooibos leaf, Phellodendri cortex or Bambusa tuldoides by discovering that the crude drug of the Rooibos leaf, the Phellodendri cortex or the Bambusa tuldoides has activities for inhibiting the production of the adiponectin at a fat cell. Various skin care preparations, the bath preparations, the food and drink and the fodder useful for the prevention, mitigation and amelioration of various disorders, the diseases, the symptoms and the like caused by the adiponectin are also provided. <P>COPYRIGHT: (C)2008,JPO&INPIT

Description

  The present invention relates to an adiponectin production inhibitor that suppresses the production of white adipocyte-derived adiponectin that functions as an adipocytokine in vivo. Furthermore, the present invention relates to a slimming skin external preparation, a slimming bath preparation, a slimming food and drink, and a slimming feed containing the adiponectin production inhibitor.

Recent advances in molecular genetics have elucidated the molecular mechanism of a new regulatory system of energy metabolism using adipocytokine as a mediator.
Adiponectin is an adipocytokine discovered in 1995 and is a protein found in mammals such as mice and humans, and is present in normal human blood at a high concentration of 5 to 10 μg / ml. Despite being specifically secreted from adipose tissue, the adiponectin concentration in blood has an inverse correlation with the visceral fat area measured by BMI and CT scan, and it is revealed that it increases with weight loss. ing. Adiponectin has an anti-arteriosclerotic effect on vascular endothelial cells, vascular smooth muscle cells, and macrophages. In addition, administration of adiponectin into the ventricle to mice confirmed a decrease in body weight through a mechanism different from that of leptin, a different adipocytokine, and revealed that adiponectin also has functions such as body weight regulation in the brain. Therefore, it is considered that adiponectin also has anti-obesity action and anti-diabetic action. In contrast to the relationship between obesity, insulin resistance and leptin resistance, adiponectin resistance is not observed in obesity and type 2 diabetes models, and therefore adiponectin replacement therapy can be expected.

Patent Document 1 discloses an extract extracted from at least one of rice bran, rahan fruit, shimeji, chrysanthemum, rye, birch and moon peach, or at least one of the plant materials or a microbial converter of these extracts. An adiponectin secretagogue that contains is shown.
However, the drug described in Patent Document 1 is a drug that promotes adiponectin secretion and does not suppress adiponectin secretion. Furthermore, the chemical | medical agent of patent document 1 does not contain the rooibos leaf of this invention, a buckwheat, and a chikujo.
JP 2005-68132 A

The physiological function of adiponectin, which is becoming clear in recent years, plays an important role in the life support mechanism of living organisms. It is expected that various lifestyle-related diseases and diseases can be prevented and improved by controlling the serum adiponectin level within an appropriate range.
However, no technique has yet been found to control adiponectin production itself.
The present invention relates to an adiponectin production inhibitor for the purpose of prevention, alleviation, and improvement of various diseases, diseases and symptoms caused by adiponectin.

In order to solve the above-mentioned problems, the present inventors consider that it is important to maintain the in vivo level of adiponectin within an appropriate range, and an adiponectin production inhibitor that acts on adiponectin production itself. Pay attention.
The present inventors have conducted extensive research on the production of adiponectin in white adipocytes with the aim of developing an adiponectin production inhibitor. As a result, it was found for the first time that a specific crude drug has an adiponectin production inhibitory action, and the present invention was completed based on this finding.

It has been found that the herbal medicines of rooibos leaves, alum and chikujo have an action of suppressing the production of adiponectin from adipocytes and are useful in various applications as an adiponectin production inhibitor.
These can be added to various skin external preparations and bath preparations including cosmetics, and foods and drinks, and are intended to prevent, alleviate and improve various diseases, diseases and symptoms caused by adiponectin. Enables the application of

The adiponectin production inhibitor of the present invention contains one or two or more herbal medicines selected from rooibos leaves, agate, and chikujo.
As the rooibos leaf used in the present invention, the rooibos leaf of the leguminous plant is used, but the above-ground part and the same genus plant can also be used.
As the buckwheat used in the present invention, the bark of the mandarin family plant yellowfin is used, but the above-ground part and the same genus plant can also be used.
The chrysanthemum used in the present invention uses a bee of a gramineous plant or an inside of a bamboo shoot, but an above-ground part or a plant belonging to the same genus can also be used.

In the present invention, as the rooibos leaf, apricot, and chikujo, the above-mentioned various parts can be used as they are or after being crushed or crushed after being dried or dried. Furthermore, the extract obtained by extracting these with a solvent, and the non-volatile content obtained by evaporating or lyophilizing the extraction solvent from the extract can be used.
Extraction solvents include water, alcohols (eg, methanol, ethanol, ethylene glycol, propylene glycol, 1,3-butylene glycol, etc.), ketones such as acetone, ethers such as diethyl ether, and aromatic carbonization such as toluene. Various solvents, such as hydrogen, are mentioned, It can use individually or in combination of 2 or more types of mixed solvents.

  The herbal medicine used in the present invention is generally used as a component of a medicine or folk medicine, and its safety has been confirmed. The adiponectin production inhibitor of the present invention can be incorporated into various skin external preparations and internal preparations such as pharmaceuticals and cosmetics, bath preparations, foods and drinks, and the like.

The adiponectin production inhibitor of the present invention contains the herbal medicines alone or as a mixture. The total blending amount of the crude drug differs depending on the dosage form, but the evaporation residue may be used as it is, and the blending amount may be appropriately adjusted depending on the intended use. The usage-amount of the adiponectin production inhibitor of this invention does not have a restriction | limiting in particular, It can adjust suitably with a use or application.
Moreover, the adiponectin production inhibitor of this invention is applicable to various forms, such as external use, internal use, and the process to a raw material. Furthermore, it can be used in combination with chemicals used in normal external use, internal use, treatment of raw materials, and the like, and thereby the effects of the present invention are more easily expressed.

The skin external preparation of the present invention contains the herbal medicine alone or as a mixture. The total amount of the crude drug varies depending on the dosage form. Moreover, there is no restriction | limiting in particular in the usage-amount of the skin external preparation of this invention, According to a user's liking, it can adjust suitably.
Moreover, the bath agent of this invention can contain the said crude drug individually or as a mixture, and the total compounding quantity changes suitably with dosage forms.
Moreover, the food and drink and the feed of the present invention can contain the herbal medicines alone or as a mixture, and the total blending amount thereof varies depending on the form.

The skin external preparation and bath preparation of the present invention include, in addition to the above-mentioned various crude drugs, known functional ingredients used in ordinary external preparations or bath preparations, such as humectants, emollients, blood circulation promoters, cell activation agents. Agents, antioxidants, anti-inflammatory agents, antibacterial agents, whitening agents, peroxide inhibitors and the like can be blended.
For example, humectants such as glycerin, butylene glycol, urea and amino acids; emollients such as squalane, macadamia nut oil, olive oil, jojoba oil and silicone oil; blood circulation promoters such as vitamin Es and pepper tincture; cell activation of nucleic acids and the like Antioxidants such as dibutylhydroxytoluene (BHT), dibutylhydroxyanisole (BHA), tocopherol acetate and ascorbic acids; anti-inflammatory agents such as glycyrrhizin and allantoin; hinokitiol, benzalkonium chloride, chlorhexidine salt, parahydroxybenzoate Various known substances such as antibacterial agents such as acid esters; whitening agents such as ascorbic acid and arbutin; peroxide inhibitors such as superoxide dismutase (SOD) can be blended.
In addition, various extracts derived from plants, animals, and microorganisms such as dragon extract, ginkgo biloba extract, peony extract, placenta extract, and lactic acid bacteria culture extract can be freely added and used.

The external preparation for skin of the present invention is an externally applicable agent, and its dosage form is not particularly limited, and examples thereof include pastes, creams, gels, ointments, lotions, emulsions, packs, powders, and haptics. it can.
The dosage form of the bath preparation of the present invention means all applicable forms, and examples thereof include solid preparations such as powders and granules, and liquid preparations such as emulsions and pastes.
The food and drink and feed dosage forms of the present invention mean all applicable forms, for example, solid preparations such as biscuits, cookies, tablets, capsules, candy, gum and powder, liquid preparations such as beverages, and jelly Examples thereof include semi-solid preparations.

In addition, in the external preparation for skin, bath preparation, food and drink of the present invention, base ingredients such as surfactants and fats and oils for the formulation, and thickeners, preservatives as necessary, Various additives such as tonicity agents, antioxidants, ultraviolet absorbers, chelating agents, fragrances, and coloring agents can be used in combination.
Although it does not specifically limit as said surfactant, A general nonionic surfactant, anionic surfactant, a cationic surfactant, and an amphoteric surfactant can be used. For example, higher alcohol alkylene oxide adduct, higher alkyl amine alkylene oxide adduct, higher fatty acid amide alkylene oxide adduct, polyhydric alcohol fatty acid ester, hydrogenated castor oil alkylene oxide adduct, polyethylene glycol sorbitan alkyl ester, sterol Nonionic surfactants such as alkylene oxide adducts such as sodium alkylsulfate, sodium alkyloxyltaurate, sodium α-olefin sulfonate, sodium polyoxyalkylene alkyl ether sulfate, alkylpyridinium chloride, Cationic surfactants such as distearyl dimerylammonium chloride; amphoteric compounds such as sodium aminopropionate and alkylpolyaminoethylglycine Surface active agents. Of these, one or more surfactants can be selected and used.

Base material components that can be used in the present invention are not particularly limited. For example, olive oil, camellia oil, avocado oil, macadamia nut oil, apricot oil, jojoba oil, squalane, squalene, horse oil, etc. And the known fats and oils.
The thickener usable in the present invention is not particularly limited, and examples thereof include polyvinyl alcohol, polyvinyl acrylamide, polyethylene glycol, and various derivatives thereof; celluloses such as hydroxyalkyl cellulose and derivatives thereof; dextran. And gums such as gelatin, gum arabic, and tragacanth; and water-soluble polymers such as carboxyvinyl polymer.
The preservative that can be used in the present invention is not particularly limited, and examples thereof include parahydroxybenzoic acid esters, paraoxybenzoic acid salts and derivatives thereof, phenoxyethanol, hinokitiol, benzalkonium chloride, chlorhexidine salts, and the like. It is done.
The isotonic agent that can be used in the present invention is not particularly limited, and examples thereof include inorganic salts such as sodium chloride, potassium chloride, and calcium chloride.
Although it does not specifically limit as an ultraviolet absorber which can be used in this invention, For example, a para amino benzoic acid, a benzophenone series ultraviolet absorber, a benzotriazole type ultraviolet absorber, etc. are mentioned.
The chelating agent that can be used in the present invention is not particularly limited, and examples thereof include ethylenediaminetetraacetic acid, phytic acid, citric acid, and water-soluble salts thereof.
In the food and drink and the feed of the present invention, various materials usually used in foods can be used.

EXAMPLES Hereinafter, although an Example and a comparative example demonstrate this invention further more concretely, this invention is not limited at all by these Examples.
Production method of plant extract About rooibos leaves, buckwheat or chrysanthemum, extract over 1 hour under reflux of 50% aqueous ethanol solution, and extract the ethanol and water extraction solvent from the obtained extract under reduced pressure. Got. These plant extracts were used in the examples shown below and subjected to an evaluation test.

[Examples 1 to 10 and Comparative Example 1]
1. Adiponectin production inhibition test using adipocytes Cell Culture Method 3T3-L1 adipocyte precursor cells were suspended in adipocyte precursor medium and seeded at 10 ⁴ per well in a 48-well plate. After culturing under conditions of 37 ° C. and 5% CO ₂ for 8 days, the medium was replaced with a differentiation induction medium (insulin 5 μg / ml, dexamethasone 1 μM, IBMX 0.5 mM) and cultured for 2 days. Next, an adipocyte medium containing 5 μg / ml of insulin was added, and after culturing for 2 days, the adipocyte medium without insulin was further cultured for 10 days.
Thereafter, after washing the cells with a new medium, the plant extracts of Examples 1 to 10 described in Table 1 or Comparative Example 1 (no addition) was added to each well so as to have the final concentration of Table 1, respectively. Cultured for 24 hours. The medium was collected from each well, centrifuged at 3500 × g, and the obtained culture supernatant was subjected to adiponectin quantification.

2. Adiponectin Quantification Method The adiponectin concentration in the culture supernatant was measured using a commercially available ELISA kit (Adiponectin ELISA Kit, manufactured by Funakoshi), and the amount of adiponectin per 1 mg protein was calculated. The measurement procedure followed the manual attached to the kit. The specific measurement procedure is as follows.
(1) Binding of adiponectin contained in the sample to the antibody plate After washing the antibody plate attached to the kit, 100 μl of each concentration of adiponectin standard solution and diluted sample (culture supernatant) is added to each well. The reaction was allowed to stand at 22 to 28 ° C. for 60 minutes.
(2) Antigen-antibody reaction After washing each well, 100 μl of biotin-labeled antibody solution was added and allowed to stand at 22-28 ° C. for 60 minutes. Further, after washing, 100 μl of enzyme-labeled streptavidin solution was added and allowed to stand at 22 to 28 ° C. for 60 minutes.
(3) Detection and quantification of adiponectin After washing each well, 100 μl of the substrate solution was added and allowed to stand at 22-28 ° C. for 15 minutes. Thereafter, 100 μl of a reaction stop solution was added, and the absorbance of each well was measured using a microplate reader set to 450 nm (measurement wavelength) and 655 nm (control wavelength). A calibration curve was created from the absorbance of the adiponectin standard solution, and the adiponectin concentration in the sample was calculated. Further, the amount of adiponectin per 1 mg protein was calculated from the obtained adiponectin concentration, and the amount of adiponectin in the system to which each plant extract was added was divided by the amount of adiponectin in the system to which no plant extract was added. The adiponectin production rate was calculated.

表１に示すように、実施例１〜１０の植物エキスはアディポネクチンの産生に対し、有意に抑制作用を示していることが判った。なお、表１において、比較例１の値は、植物エキスが無添加の時の値（標準値）である。

As shown in Table 1, it was found that the plant extracts of Examples 1 to 10 significantly inhibited the production of adiponectin. In Table 1, the value of Comparative Example 1 is a value (standard value) when no plant extract is added.

  Next, although the formulation example of the skin external preparation for slimming of this invention which mix | blended the said plant extract is shown, these examples do not limit this invention at all.

Formulation examples of slimming skin external preparations Slimming skin external preparations were prepared with the compositions shown in Tables 2-5. The preparation method is as follows. The “remaining amount” of water in the table means an amount for making the total amount 100 g.

Ａ成分、Ｂ成分ともに８０℃で加温溶解し、Ｂ成分をＡ成分に撹拌しながら徐々に加え乳化した。撹拌しながら冷却し、４０℃でＣ成分を添加し、３５℃で調製を終了した。 [Examples 11 to 15 and Comparative Example 2 (skin lotion)]

Both the A component and the B component were dissolved by heating at 80 ° C., and the B component was gradually added to the A component while stirring to emulsify. The mixture was cooled with stirring, the C component was added at 40 ° C, and the preparation was completed at 35 ° C.

Ｄ成分、Ｅ成分のそれぞれを８０℃に加温溶解した。Ｅ成分を撹拌しながらＤ成分を加え均一とした後、ホモミキサーで乳化した。ハンドルミキサーで撹拌しながら冷却し、４０℃以下でＦ成分を添加し、調製した。 [Examples 16 to 19 and Comparative Example 3 (milky lotion)]

Each of component D and component E was heated and dissolved at 80 ° C. The D component was added and homogenized while stirring the E component, and then emulsified with a homomixer. It cooled, stirring with a handle mixer, and added F component at 40 degrees C or less, and prepared.

Ｇ成分、Ｈ成分をそれぞれ７５℃に加温した。パドルミキサーでＧ成分を撹拌しながら、Ｈ成分を少量ずつ加えた。撹拌しながら冷却し、４０℃でＩ成分を添加した後、約３５℃で調製した。 [Examples 20 to 23 and Comparative Example 4 (cream)]

G component and H component were each heated to 75 ° C. While stirring the G component with a paddle mixer, the H component was added little by little. After cooling with stirring and adding component I at 40 ° C, it was prepared at about 35 ° C.

Ｊ成分、Ｋ成分をそれぞれ７５℃に加温した。パドルミキサーでＪ成分を撹拌しながら、Ｋ成分を少量ずつ加えた。その後、パドルミキサーで撹拌しながら冷却し、４５℃以下でＬ成分を添加して調製した。 [Examples 24 to 30 and Comparative Example 5 (body cream)]

The J component and the K component were each heated to 75 ° C. While stirring the J component with a paddle mixer, the K component was added little by little. Then, it cooled, stirring with a paddle mixer, and prepared by adding L component at 45 degrees C or less.

Evaluation of slimming skin external preparations For the slimming skin external preparations of Examples 11 to 30 and Comparative Examples 2 to 5, 48 males and 48 females who suffer from the fat layer of the waist and hips were monitored, and fat was removed. The increase and decrease of layers were investigated. That is, it was divided into 24 groups of 2 males and 2 females, and the slimming skin external preparations shown in Table 6 were used respectively. In addition, the test method for the external preparation for skin is to take each skin external preparation in a size of about 500 yen coins on the palm after daily bathing, and to conduct a normal life except for applying it while massaged. Continued for a while. The waist and hip dimensions before and after the test were measured to evaluate the slimming effect. Table 6 shows the measurement results.

アディポネクチン産生抑制作用を有する植物エキス、つまり本発明のアディポネクチン産生抑制剤を添加していない比較例２〜５の皮膚外用剤では、ウエストおよびヒップの寸法はほぼ変化しなかったが、本発明のアディポネクチン産生抑制剤を添加した痩身用皮膚外用剤はいずれも寸法が減少し、痩身効果が認められた。

In the plant extract having an adiponectin production inhibitory action, that is, the skin external preparations of Comparative Examples 2 to 5 to which the adiponectin production inhibitor of the present invention was not added, the dimensions of the waist and hips were not substantially changed, but the adiponectin of the present invention The slimming skin external preparations to which production inhibitors were added all decreased in size and a slimming effect was observed.

  Next, although the formulation example of the slimming bath preparation of this invention which mix | blended the said plant extract is shown, these examples do not limit this invention at all.

Ｍ成分、Ｎ成分をそれぞれ均一になるまで混合した後、Ｍ成分とＮ成分とを混合し、さらに均一になるまで充分混合した。 [Examples 31 to 39 and Comparative Example 6]

The M component and the N component were mixed until they became uniform, and then the M component and the N component were mixed and mixed well until they became more uniform.

Evaluation of slimming bath agent The slimming bath agent of the above-mentioned prescription example was monitored for 20 men and 20 women to examine the feeling of use. That is, it was divided into 10 groups of 2 men and 2 women, and the slimming bathing agents shown in Table 8 were used at the time of bathing. The test method for the bathing agent was a normal life except that 30 ml of the bathing agent was dissolved in 180 L of hot water at the time of daily bathing, mixed well and bathed, and this was continued for one month. One month later, the feeling of use was evaluated in the following four stages for each part of the upper arm, waist, hips, and thighs. The evaluation is shown in Table 8.
○: Feeling tightened △: Feeling slightly tightened ×: Not changed XX: Feeling slack

  In the plant extracts having an adiponectin production inhibitory action, that is, the bath preparations of Examples 31 to 39 to which the adiponectin production inhibitor of the present invention was added, there were many evaluations of a tight feeling and a slightly tight feeling regarding the upper arm, waist, hip and thigh. Since most of the evaluation that the bath preparation of Comparative Example 6 to which the adiponectin production inhibitor of the present invention was not added was the same, the slimming bath preparation to which the adiponectin production inhibitor of the present invention was added Was also significant.

  Next, examples of prescriptions for slimming food and drink and feed according to the present invention containing the plant extract are shown, but these examples do not limit the present invention in any way.

Ｐ成分を全部混ぜ合わせ、４０℃に加温したＱ成分とＲ成分を混ぜ合わせ、均一になった時点でよくこねた。３５℃で４０分一次発酵させ、空気を抜くように再度こねた。４０℃で４０分二次発酵後、１８０℃で２５分間焼いた。 Slimming food and drink formulation examples [Examples 40 to 42 and Comparative Example 7 (bread)]

All of the P component was mixed, and the Q component and R component heated to 40 ° C. were mixed, and kneaded well when it became uniform. Primary fermentation was carried out at 35 ° C. for 40 minutes, and kneaded again so as to remove air. After secondary fermentation at 40 ° C. for 40 minutes, it was baked at 180 ° C. for 25 minutes.

Ｓ成分を、予めＴ成分をＵ成分に溶解した８０℃のお湯に溶かし、よく混ぜ合わせた。型に１０ｇずつ流し入れた後冷蔵庫で固めた。 [Examples 43 to 45 and Comparative Example 8 (jelly)]

The S component was dissolved in 80 ° C. hot water in which the T component was previously dissolved in the U component and mixed well. After pouring 10 g into the mold, it was hardened in the refrigerator.

Ｖ成分とＷ成分を良く混ぜ合わせ、冷蔵庫で冷やした。塩抜きした羊の腸に、混ぜ合わせたものを詰めて、それを６０〜７０℃で１時間燻製にし、さらに１時間かけて乾燥させた。乾燥させたものを７０℃程度のお湯で３０分程度ゆで、殺菌処理した。再度、乾燥させたものを完成品とした。 [Examples 46 to 48 and Comparative Example 9 (sausage)]

V component and W component were mixed well and cooled in a refrigerator. The salted sheep intestines were filled with the blend, smoked at 60-70 ° C. for 1 hour, and further dried for 1 hour. The dried product was sterilized by boiling in hot water at about 70 ° C. for about 30 minutes. The dried product was again used as the finished product.

Ｘ成分とＹ成分を良く混ぜ合わせる。混ぜ合わせたものを打錠機を用いてタブレット状に押し固め、錠剤を形成した。 [Examples 49 to 51 and Comparative Example 10 (Supplement (Tablet))]

Mix X component and Y component well. The mixture was pressed into a tablet using a tableting machine to form a tablet.

Evaluation of slimming foods and feeds Regarding the slimming foods and feeds of the above-mentioned prescription examples, 32 men and 32 women who suffered from the fat layer of the waist and hips were monitored to examine the increase and decrease of the fat layer. . That is, it divided into 16 groups of 2 men and women, and used the food and drink shown in Table 13, respectively. In addition, the test method of this food / drink was carried out for one month, carrying out a normal life except ingesting each food / beverage after every dinner. The waist and hip dimensions before and after the test were measured to evaluate the slimming effect. Table 13 shows the measurement results.

アディポネクチン産生抑制作用を有する植物エキス、つまり本発明のアディポネクチン産生抑制剤を添加していない比較例７〜１０の飲食物では、ウエストおよびヒップの寸法はほぼ変化しなかったが、本発明のアディポネクチン産生抑制剤を添加した痩身用飲食物ではいずれも寸法が減少し、痩身効果が認められた。

In the plant extract having an adiponectin production inhibitory activity, that is, the food and drink of Comparative Examples 7 to 10 to which the adiponectin production inhibitor of the present invention was not added, the waist and hip dimensions were not substantially changed, but the adiponectin production of the present invention was not changed. The slimming foods and drinks to which the inhibitor was added all decreased in size, and a slimming effect was observed.

  The herbal medicine of the present invention has an action of suppressing the production of adiponectin and is useful as an adiponectin production inhibitor. The adiponectin production inhibitor of the present invention can be incorporated into cosmetics, quasi drugs and bath preparations. In particular, as a slimming skin external preparation, slimming bath preparation, slimming food and drink, and slimming feed, production of adiponectin can be suppressed systemically or locally, and obesity can be improved, suppressed or prevented.

Claims (4)

  An adiponectin production inhibitor containing one or more herbal medicines selected from rooibos leaves, apricots and chikujo.   A slimming skin external preparation containing the adiponectin production inhibitor according to claim 1.   A slimming bath preparation comprising the adiponectin production inhibitor according to claim 1. A slimming food and drink and a slimming feed comprising the adiponectin production inhibitor according to claim 1.