JPH1171290A - Hyaluronidase inhibitor and food and beverage containing the same - Google Patents

Abstract

PROBLEM TO BE SOLVED: To obtain a hyaluronidase inhibitor improved in inhibitory effects and high in safety, useful as a therapeutic agent for rheumatism and deformant arthritis, containing an extract of Phyllantus emblucus as an active ingredient. SOLUTION: This inhibitor comprises an extract of Phyllantus emblucus as an active ingredient. The extract is obtained, for example, by crushing and grinding the fruit, seed, bark, etc., of amla and extracting the ground material with a solvent such as water at 40-100 deg.C for 10 minutes to 24 hours. The hyaluronidase inhibitor is added to a food and beverage such as confectionery, bread, grain, noodles to provide a hyaluronidase inhibiting food and beverage. In this case, the amount of the hyaluronidase added to a food and beverage is preferably 0.01-5 w/w % based on the food and beverage calculated as the amla extracted essence.

Description

DETAILED DESCRIPTION OF THE INVENTION

[0001]

BACKGROUND OF THE INVENTION 1. Field of the Invention The present invention relates to a novel hyaluronidase inhibitor comprising a highly safe plant extract and a food or drink for inhibiting hyaluronidase containing the hyaluronidase inhibitor.

[0002]

2. Description of the Related Art Hyaluronic acid is a linear high-molecular polysaccharide formed by alternately binding β-DN-acetylglucosamine and β-D-glucuronic acid, and has a large molecular weight of 10%.
More than 100,000 are known. Hyaluronic acid is widely distributed in connective tissues and the like of mammals, and is considered to contribute to the retention of intercellular water in the skin and the maintenance of lubricity and flexibility of the skin. Moreover, hyaluronidase is widely distributed in the human body, and hyaluronic glucosaminidase is known as hyaluronidase in mammals including humans. On the other hand, it is known that hyaluronic acid in joint cavities of patients with rheumatism and osteoarthritis is reduced in molecular weight, and a hyaluronidase inhibitor that suppresses the degradation of hyaluronic acid is expected as a therapeutic agent for such diseases. I have. Furthermore, it is said that hyaluronidase is associated with the occurrence of inflammation, and it is considered that inhibitors of this enzyme are also effective in preventing and treating inflammation and allergy. Already, various hyaluronidase inhibitors have been extracted from various plants, and patent applications have been filed [Japanese Patent Application Laid-Open No. 1-128933 (Shaku-oh, Uren etc.),
JP-A-2-11520 (Jiyu, Gennoshoko etc.), JP-A-2-221229 (licorice etc.), JP-A-6-80576 (Chippi, pheasant etc.)].

[0003]

As described above, hyaluronidase inhibitors are expected to be promising not only for the treatment but also for the prevention of diseases. Great care must be taken.
An object of the present invention is to search for a new hyaluronidase inhibitor having a high inhibitory effect and high safety from food materials,
An object of the present invention is to provide the inhibitor and a food or drink for inhibiting hyaluronidase containing the same.

[0004]

SUMMARY OF THE INVENTION To summarize the present invention, a first invention of the present invention relates to a hyaluronidase inhibitor comprising an extract of Filanthus emblica as an active ingredient, and a second invention relates to the inhibitor of the invention. And a food or beverage for hyaluronidase inhibition.

As a result of intensive studies, the present inventors have found that an extract having a scientific name of Phylanthus emblica has a high hyaluronidase inhibitory activity, and have completed the present invention.

[0006]

BEST MODE FOR CARRYING OUT THE INVENTION The present invention will be specifically described below. First, Filanthus emblica in the present invention is native to India, Ceylon, the Malay Peninsula, Sunda Island, and China, and is a deciduous small tree cultivated in part of Taiwan and belongs to the Euphorbiaceae family. Filanthus emblica has been eaten raw for a long time in India and other countries, and has been used as a food and drink such as jelly and pickles. It is used for diarrhea, cholera, gallbladder, etc., and is extremely safe for the human body. In addition, Yukan in the Japanese name and those commonly called Amala are also used in the present invention.
Belongs to Emblica.

[0007] As a raw material of Filanthus emblica for extraction of the present invention, fruits, seeds, bark and the like of Amara can be used. The shape of the fruit, seed, bark, etc. to be used is not particularly limited, but it is preferable to use it after being crushed or pulverized from the viewpoint of extraction efficiency.

[0008] The extraction method from the Amara raw material is not particularly limited, and examples thereof include a commonly used method such as a batch method and a column method. Amara fruit, seeds, the solvent used for the extraction of the active ingredient from the bark, etc., water, methyl alcohol, ethyl alcohol, butanol, ethyl acetate, ether, acetone or one or more solvents such as n-hexane and the like. They can be used by appropriately mixing. Although the extraction temperature is not particularly limited, a temperature of about 40 to 100 ° C is preferable, and when an organic solvent is used, 50 to 90 ° C is preferable. Also, the extraction time is not particularly limited, but is several minutes to several tens of hours, preferably 10 minutes to 24 hours. Furthermore, the presence or absence of stirring at the time of extraction can be arbitrarily selected.

The extract of the present invention is used as a hyaluronidase inhibitor as it is, or in a diluted or concentrated state, or in the form of a concentrated, dried powder, granule or tablet. Thereby, a food or beverage for hyaluronidase inhibition can be provided. Foods and drinks here include, for example, confectionery and baking, flour and noodles, agricultural and forest processed foods, livestock processed products, milk and dairy products, oils and fats processed oils, alcoholic beverages, beverages, seasonings, food ingredients, etc. Is mentioned.

The amount of the hyaluronidase inhibitor to be contained in the food or drink is 0.001 w / w% per food or drink in terms of the amara extract (solid) obtained in Example 1 described later.
As long as it is at least, preferably 0.01 to 5 w / w%
It is.

[0011]

EXAMPLES Hereinafter, the present invention will be described more specifically based on examples, but the present invention is not limited to these examples.

Example 1 (Preparation of Hyaluronidase Inhibitor by Amara Extract) Five-fold amount of distilled water was added to 200 g of dried and ground Amara fruit, heated and extracted with hot water at 90 ° C. for 1 hour. The supernatant and the residue were separated. The supernatant obtained by washing the residue with warm water is combined with the above-mentioned supernatant, and is subjected to 50 ° C. under reduced pressure (720 mmHg) according to a conventional method.
And dried to obtain 30 g of Amara fruit extract (hereinafter abbreviated as Amara extract).

Example 2 (Results of Measurement of Hyaluronidase Inhibiting Activity) The Amara extract obtained in Example 1 was dissolved in a 0.1 M phosphate buffer solution (pH 4), and 2 mg / ml [specific gravity: 1.
0 (20 ° C.)] for activity measurement. Hyaluronidase inhibitory activity was measured by Morgan-Elson.
-Elson) method [The Journal of Biological
Chemistry (J. Biol. Chem), Vol. 217, p. 959 (1955)].
That is, 1.9 mg / m of beef testicular hyaluronidase [Type NS, manufactured by Sigma-Aldrich Japan Co., Ltd.]
0.1M phosphate buffer (pH 4.0)
To 1, 0.2 ml of the above extract sample was added in a stepwise change in concentration. After incubating the mixture for 20 minutes at 37 ° C, 0.2 ml of a phosphate buffer containing 0.5 mg / ml of Compound 48/80 (manufactured by Sigma-Aldrich Japan) was added, and the mixture was added at 37 ° C.
Incubated for 0 minutes to activate the enzyme. Further, a phosphate buffer solution containing 0.8 mg / ml of potassium hyaluronate (manufactured by Wako Pure Chemical Industries, Ltd.) as a substrate was used.
Then, an enzyme reaction was carried out at 37 ° C. for 40 minutes. Then, 0.2 ml of 0.4N sodium hydroxide and 0.2 ml
ml of 0.8 M sodium borate buffer (pH 9.2)
And boil for 3 minutes to inactivate the enzyme, then 1 w / v
% P-dimethylaminobenzaldehyde-containing 10N hydrochloric acid: glacial acetic acid mixture [1.25: 98.75 (v: v)]
Was added and the color reaction was carried out at 37 ° C. for 20 minutes, and the acetyl amino acid released from hyaluronic acid was quantified by the color reaction. According to this method, the inhibition by the extract is measured as a decrease in absorbance, and the inhibition rate is calculated by the following formula (Equation 1).
It is represented by

[0014]

## EQU1 ## Inhibition rate (%) = [(absorbance of blank−absorbance of sample) / (absorbance of blank)] × 100

The results are shown in FIG. The concentration of the Amara extract was expressed as a ratio of the solid content weight of the Amara extract to the reaction solution volume. In FIG. 1, the vertical axis indicates the inhibition rate (%), and the horizontal axis indicates the concentration (μg / ml). 50 of the Amara extract obtained in Example 1 obtained from FIG.
The concentration (1C ₅₀ ) showing% inhibition is 175 μg / ml. Tencha extract, a hyaluronidase inhibitory activity and an antiallergic agent, which has been known and put to practical use,
Alternatively, the values were almost equivalent to those of 200 μg / ml or 100 μg / ml of apple polyphenol. The Amara extract used this time is a crude extract directly extracted with hot water from Amara fruit, and has a very strong inhibitory effect as such a crude extract.

Example 3 (Preparation of tablet) Using the Amara extract obtained in Example 1, a tablet composition was prepared by a conventional method based on the composition shown in Table 1, and the tablet was prepared using a tablet press. Was prepared.

[0017]

[Table 1]

The above composition is 40 mg per tablet (2 g).
(2 w / w%) Amara extract extract, and can be expected to have effects such as improvement of allergy.

Example 4 (Preparation of tofu) 1 g of soymilk cooled to 15-18 ° C., 4 g of natural bittern, and the Amara extract 13 obtained in Example 1
After adding 3mg, pack into plastic cups,
Sealed with sealer. This was heated in hot water (85 ° C.) for 45 minutes to coagulate, and then cooled on ice to obtain a filled silken tofu. This tofu is 40 mg (0.4 g) per one chop (300 g).
013 w / w%), and the texture and taste were good.

Example 5 (Preparation of candy) 1.2 kg of sugar and 0.8 kg of starch syrup were dissolved in a dissolver (11
0 ° C) and then mixed with a cooker at 120-130 ° C.
The mixture was boiled until the water content was reduced to 2% or less, 8 g of Amara extract extract and perfume were kneaded and mixed well, then cooled and molded. This candy is 40mg per grain (4g)
(1% w / w) Amara extract, and both flavor and taste were good.

Example 6 (Preparation of Beverage) Raw materials containing the Amara extract obtained in Example 1 in Table 2 were mixed to obtain a beverage containing the Amara extract.

[0022]

[Table 2] (*: Contains 25 w / w% water)

The above raw materials were dissolved in water, filled up to 1 liter, sterilized (80 ° C., 10 minutes), and bottled. 100 ml of this beverage contained 40 mg (0.04 w / w%) of the Amara extract, and both flavor and taste were good.

Example 7 (Preparation of ramen) 400 g of flour exclusively for ramen (wheat flour containing brine etc.)
Then, 160 mg of Amara extract extract obtained in Example 1 and 160 ml of water were added thereto, and mixed so as to be ragged. This was noodle-made with a home noodle-making machine (manufactured by Sanyo Electric Co., Ltd.) to obtain ramen. This ramen is served alone (flour 1
40 mg (0.04 equivalent) of Amara extract
w / w%), and both texture and taste were good.

[0025]

As described above in detail, the extract of Amara belonging to Filanthus emblica has a high hyaluronidase inhibitory activity. Amara is a highly safe substance that has been used as a crude drug or food and drink since ancient times, and the present invention is a hyaluronidase inhibitor that is extremely effective in practical use. In addition, by incorporating this hyaluronidase inhibitor into foods and drinks, foods and drinks for inhibiting hyaluronidase with good quality such as flavor can be easily obtained.

[Brief description of the drawings]

FIG. 1 is an explanatory diagram showing the hyaluronidase inhibitory activity of an Amara extract.

Continued on the front page (51) Int.Cl. ⁶ Identification code FI A23L 2/52 A23L 2/38 C 2/38 C12N 9/99 C12N 9/99 A23L 2/00 F (72) Inventor Tatsuya Kawabe Otsu, Shiga 3-4-1, Seta-ichi, Takara Shuzo Co., Ltd. Central Research Laboratory (72) Inventor Hideo Morita 3-4-1, Seta, Otsu-shi, Shiga Prefecture Takara Shuzo Co., Ltd. Central Research Laboratory Co., Ltd.

Claims (2)

[Claims] 1. A hyaluronidase inhibitor comprising an extract of Filanthus emblica as an active ingredient. 2. A food or drink for inhibiting hyaluronidase, comprising the hyaluronidase inhibitor according to claim 1.