JP3178822B2 - 3α-hydroxysteroid dehydrogenase inhibitory activator and hyaluronidase inhibitory activator - Google Patents

Description

Description: TECHNICAL FIELD The present invention relates to an anti-inflammatory composition effective for hay fever, and more particularly, to a composition which can impair the flavor of food even when added to food. The present invention relates to a 3α-hydroxysteroid dehydrogenase inhibitory activity and a hyaluronidase inhibitory activity having a good taste.

[Conventional technology]

In recent years, the prevalence of allergic diseases such as hay fever,
It tends to increase significantly. Especially in Japan, the number of cedar pollinosis patients is increasing rapidly, and it is estimated that about 10% of urban residents have cedar hay fever. Pollinosis is characterized by the onset of pollen only when it is flying.However, once affected, it causes unpleasant initial symptoms such as persistent itching and constant runny nose. It generally lasts weeks to months until it decreases. Also, as these early symptoms progress, headaches, gastrointestinal disorders,
Fatigue and the like due to insomnia are also occurring, which has developed into a serious social problem.

Therefore, research on the treatment of hay fever has been widely conducted, and medical products have been actively developed. Not only therapeutic drugs at hospitals, but also over-the-counter drugs such as antihistamines and corticosteroids are widely used.

However, the above-mentioned over-the-counter medicines can be easily used by patients, but if used improperly, they may cause drowsiness or cause corneal infection.

Therefore, research on Kampo therapy using crude drugs has recently attracted attention. Kampo therapy is a treatment using a crude drug derived from natural products, and although it is slightly delayed, it can be used to improve not only local symptoms but also physical condition, so the medicinal effect is systemic. is there.

In addition, there are few side effects compared to drugs, and the safety is high even when taken for a long time.

The Kampo medicines used in the Kampo therapy include Kakkonto,
Small seiryu hot water and Eppika hot water are known. These herbal medicines are composed of several kinds of crude drugs. For example, in the case of Kakkonto, crude drugs such as kakkon, mao, cinnamon bark, peony, jujube, licorice, ginger and the like are included.

These herbal medicines are used in the form of granules, tablets, and the like, but often suffer from peculiar bitterness and poor aftertaste, and have a drawback that they are difficult to take. Therefore, the research was started on the idea that providing the herbal components in Chinese medicine in the form of foods such as gum, candy, and beverages would make it easier to take.

However, some of these crude drugs have not been approved as food additives. In addition, it has been found that even if it can be used for foods, it often has a peculiar smell and taste, thereby impairing the original flavor of the foods.

[Problems to be solved by the invention]

The present invention has been made in view of such circumstances, and an object of the present invention is to provide 3α-hydroxysteroid dehydrase.
ogenase) or hyaluronidase
3α-hydroxysteroid dehydrogenase inhibitory activity that is excellent in anti-inflammatory effect against hay fever, high in safety, and does not impair the flavor of food even when added to food. Agents and hyaluronidase inhibitory active agents.

[Means for solving the problem]

An object of the present invention is to provide a 3α-hydroxysteroid dehydrogenase (3α-hydroxystease) having at least one food material powder or extract selected from the following group, which is excellent in anti-inflammatory effect against hay fever.
a) All spices, basil, coriander, nutmeg, pepper, sage, thyme, hops, gloves (cloves), cinnamon (cinnamon), mint (mint), licorice (licorice), fragrance, rosemary At least one spice or herb selected from the group consisting of perilla, perilla, asenyaku, chrysanthemum, and gardenia.

b) Coffee beans c) Cocoa beans d) Chlorella e) Mushrooms f) Rosaceae fruits g) Non-fermented tea h) Propolis and powder or extract of at least one food material selected from the following group Hyaluronidase (hy
aluronidase).

a) All spices, basil, coriander, sage,
At least one spice or herb selected from the group consisting of thyme, hops, gloves (cinch), cinnamon (cinnamon), mint (mint), licorice (licorice), fenca, rosemary, perilla, asenyaku, chrysanthemum, and gardenia.

b) Coffee beans c) Cocoa beans d) Chlorella e) Mushrooms f) Rosaceae fruits g) Propolis That is, the present inventors have developed an anti-inflammatory function from various food materials conventionally used in foods. We considered whether there was a method of screening food materials having the above and examined. As a result, it has been noted that it has been reported that the higher the inhibitory activity rate against 3α-HSD or HYD, the higher the anti-inflammatory effect (Hitomi et al .; Abstracts of the 34th Annual Meeting of the Japanese Society of Pharmacognostics 16 (1987)) , Kakegawa et al., Proceedings of the 14th Symposium on Crude Drug Analysis, P.35 (1985)). When this method was applied to food materials, certain food materials showed strong inhibitory activity against 3α-HSD or HYD. Further, as a result of eating these food materials actually added to the food, it was confirmed that a food having an excellent anti-inflammatory effect and a good flavor was obtained, and the present invention was completed.

 Next, the present invention will be described in detail.

First, 3α-HSD is NAD as shown in the following formula (I).
P or NAD-related oxidoreductase. This enzyme is strongly inhibited by existing anti-inflammatory agents, and a high correlation has been observed between the inhibitory concentration and anti-inflammatory results in vivo.

HYD, as shown in (II) below, is an enzyme that degrades hyaluronic acid, is strongly inhibited by existing anti-inflammatory agents, and has a high correlation between the inhibitory concentration and the anti-inflammatory effect in vivo. Has been recognized.

The 3α-hydroxysteroid dehydrogenase inhibitor and the hyaluronidase inhibitor of the present invention are selected from the following spices or herbs, coffee beans, cocoa beans, chlorella, mushrooms, rose fruits, unfermented tea, and propolis. And at least one food material powder or extract.

First, the spices or herbs used in the present invention are mainly spices obtained from dried seeds, fruits, flowers, buds, leaf stems, bark, roots, etc. of plants grown in the tropical to temperate regions, and are characterized by medicinal properties or dishes. Herbs with seasonal flavor (including those called spices in the classification, but whole plants, leaves, flowers, fruits, rhizomes, etc.) are used. For example, allspice, basil, coriander, nutmeg (mace), pepper (black pepper, white pepper), sage, thyme, hops, gloves (cloves), cinnamon (cinnamon), mint (mint), licorice (licorice), fenka , Rosemary, perilla, asenyaku, chrysanthemum flower, gardenia, and the like.

Next, the coffee beans used in the present invention are roasted green coffee beans, and varieties are not particularly limited.

The cocoa beans used in the present invention are seeds produced in a tropical evergreen tree, and include those that are naturally fermented and dried, and processed products thereof, such as cocoa and chocolate.

The chlorella used in the present invention is a single-celled green alga belonging to the order Chlorococcus, and may be a powdered algal body or one extracted with a suitable solvent after cell wall destruction.

Next, mushrooms used in the present invention include shiitake mushroom, enoki mushroom, shimeji mushroom, and the like.

In addition, the fruits of the Rosaceae used in the present invention are Yamako,
Loquat, apricot, plum and the like can be mentioned. For example, plum is used in the form of fresh fruits or processed products such as dried plum, pickled plum, plum juice and the like.

Examples of the unfermented tea used in the present invention include green tea, kairi tea, puer tea, hoji tea, and brown rice tea.

Further, the propolis used in the present invention is a sticky substance obtained by applying a resinous substance collected from various plants in the home range of a bee to a hive, and is one of the bee products having a complex composition. is there. Examples of the form to be used include natural propolis, its purified product and its processed product.
In particular, propolis natural powder, propolis tincture, propolis extract powder, etc. described in JP-A-62-210952 are preferably used as purified products.
Propolis natural powder is obtained by disinfecting, purifying, and powdering natural propolis, and contains 50 to 55% of a propolis component, 5 to 10% of essential oil, about 5% of pollen, and the like. Propolis tincture is tincture (alcohol solution) obtained by extracting natural propolis with alcohol, and has a solid content adjusted to 10%. Propolis extract powder is a water-dispersible powder obtained from the above-mentioned propolis tincture by a method of controlling the crystallization rate of a hydrophobic portion by adding sugar.

The above-mentioned form of the food material as an inhibitor for 3α-HSD and HYD may be a powder of a dried food material, an essential oil obtained by pulverizing these and performing steam distillation or the like, or An extract obtained by extracting a food material with a solvent such as water or ethanol may be used.

These may be used alone or in combination of several kinds. In particular, it is preferable to use an organic acid such as citric acid or fruit juice such as apple, peach, strawberry, orange, grape or the like containing a large amount of organic acid together with these, because a synergistic effect on inflammation is recognized.

These food materials contain, for example, components as shown in Table 1. Table 2 shows the inhibitory activity rates for these 3α-HSD and HYD.

Next, foods to which these 3α-hydroxysteroid dehydrogenase inhibitory activators and hyaluronidase inhibitory activators are added include gums, candy, jellies, tablet confections, beverages and the like. In particular, gums, candy, and tablet confections are foods that are eaten for a long time in the oral cavity, so that the 3α-hydroxysteroid dehydrogenase inhibitory activator and the hyaluronidase inhibitory activator gradually elute, making it easier to work on inflamed sites. This is preferable.

Also, the method of addition is not particularly limited, and may be prepared by mixing with other raw materials according to a conventional method.

Further, the 3α-hydroxysteroid dehydrogenase inhibitor and the hyaluronidase inhibitor are
Since it is relatively stable to heat, it can be used for foods heated at high temperatures such as heated beverages, snacks and cakes.

In addition to the above-mentioned foods, it is preferable to use it as a mouthwash, because it can be used as a mouthwash without discomfort in the mouth.

〔The invention's effect〕

As described above, the 3α-hydroxysteroid dehydrogenase inhibitor and the hyaluronidase inhibitor of the present invention have an excellent anti-inflammatory effect on hay fever. Moreover, since the food material is used as a raw material, the flavor of the food is not impaired and the safety is high.

Also, since it is stable against heat, it can be made into various forms of food.

 Next, the present invention will be specifically described based on examples.

[Examples 1 to 12 and Comparative Examples 1 and 2] Board gums and candy were prepared with the compositions shown in Tables 4 and 5, respectively, according to a conventional method. In addition, the thing which does not use an anti-inflammatory food material in the table | surface was set to Comparative Examples 1-2. Then these plate gums,
The candy was measured for its inhibitory activity against 3α-HSD and HYD, and further subjected to a sensory evaluation. In addition, the inhibitory activity rate and the sensory evaluation were performed as follows.

(1) according to the method of 3α-HSD inhibitory activity rate Hitomi et al. (Japan herbal Society 34th Abstracts 16
(1987) 3α-HSD inhibitory activity was measured.

1) Preparation of sample A 3α-HSD crude enzyme solution was prepared by the method shown in FIG.
It was diluted to 15% with M phosphate buffer (pH 6.0).

The food sample solution was prepared as follows. First, 5.0 g of plate gum cut into about 5 to 10 mm was extracted with stirring at 4 ° C. for 5 hours together with 100 ml of the above buffer solution. In addition, the candy was dissolved or suspended in the above-mentioned buffer to prepare 50 mg / ml.

2) Inhibitory activity measurement method In 0.1 M phosphate buffer (measured concentration 40 mg / ml) (pH 6.0)
At 37 ° C, 2.0 ml of food sample solution and 0.2 ml of 15% 3α-HSD crude enzyme solution
Was shaken for 10 minutes and 0.2 ml of NADPH (135 μM) was added. Ten
After shaking for minutes, the substrate nitroacetophenone (Nitroa
The reaction was started by adding 0.1 ml of cetophenone (750 μM), and the decrease in NADPH after 10 minutes was measured at an absorbance of 340 nm to calculate the inhibition rate.

 The concentrations in parentheses are final concentrations.

On the other hand, the control system was prepared by changing the food sample solution to the above phosphate buffer, and the substrate nitroacetophenone (Nitr
oacetophenone) was replaced with the above-mentioned phosphate buffer, and used as a blank system, and the amount of NADPH reduction was determined in the same manner.

 The inhibition rate is determined by the following equation.

C: NADPH decrease at 340 nm in the control system (partially described as △ 340 below) CB: △ 340 in the control system blank S: △ 340 in the sample system SB: △ 340 in the sample system blank (2) HYD inhibitory activity Rate 1) Preparation of sample First, 5.0 g of board gum cut into about 5 to 10 mm was mixed with 50 ml of 0.1 M acetate buffer (pH 3.5) at 4 ° C.
And extracted for 5 hours. In addition, the candy was dissolved or suspended in the above acetate buffer to prepare 400 mg / ml.

2) Inhibitory activity measurement method 0.2 ml of food sample solution at 37 ° C in 0.1 M acetate buffer (pH 3.5)
And 0.1 ml of a HYD solution (9 mg / ml 800NFunit / mg manufactured by Sigma) were shaken for 20 minutes, and then 0.2 ml of a 12.5 mM CaCl ₂ solution was added, followed by shaking for 20 minutes.
In addition, the substrate potassium hyaluronate (1.2 mg / ml) 0.5 ml
Was added to start the reaction. Forty minutes later, 0.4N sodium hydroxide was added to neutralize. The resulting oligosaccharide having N-acetylglucosamine at the end (NAG) was converted to Morgan-Elson (M
organ-Elson) at 585 nm.

The control system was prepared by changing the food sample solution to the above acetate buffer, and the blank system was prepared by changing the substrate to the above acetate buffer, and the amount of NAG produced was determined in the same manner.

 The inhibition rate is determined by the following equation.

C: Amount of NAC generated at 585 nm in the control system (partially described as $ 585) CB: $ 585 in the control system blank S: $ 585 in the sample system SB: $ 585 in the sample system blank (3) Sensory evaluation The taste and anti-inflammatory effect of eating the plate gum and candy of the example and the comparative example in the mouth twice a day (once every 5 minutes, 4 hours) for 3 days using 20 panelists. Sensory evaluation.

The panelists were 20 adults of our laboratory who were aware of any of the initial symptoms of hay fever such as runny nose, stuffy nose, sneezing, and itchy eyes. The experiment was conducted in early April.

 The results are shown in Tables 4 and 5.

From the above results, all of the examples were 3α-HSD, HY
It showed a strong inhibitory activity against D, and the panelists also recognized the effect on inflammation in eating results.

In addition, the flavor was as good as that of the comparative example, and the flavor was good.

Therefore, the example product to which the 3α-hydroxysteroid dehydrogenase inhibitory activity and the hyaluronidase inhibitory activity were added was able to reduce the symptoms of hay fever without affecting the flavor.

In contrast, all of the comparative examples had a low inhibitory activity rate against 3α-HSD and HYD, and had a low effect on the eating results of panelists.

[Brief description of the drawings]

FIG. 1 is an explanatory view showing a method for preparing a crude 3α-HSD enzyme solution in Examples.

──────────────────────────────────────────────────続 き Continued on the front page (51) Int.Cl. ⁷ Identification symbol FI A61P 29/00 A61P 29/00 Joint Panel Referee Yasuo Yoshimura Referee Kazuko Miyamoto Referee Hiroshi Fukatsu (56) References 185267 (JP, A) JP-A-1-181750 (JP, A) JP-A-1-165595 (JP, A) JP-A-64-47721 (JP, A) JP-A-63-198693 (JP, A) JP-A-61-186325 (JP, A)

Claims (2)

(57) [Claims] 1. A 3α-hydroxysteroid dehydrogenase (3α-hydroxysteroid) having an excellent anti-inflammatory effect against hay fever, characterized by containing a powder or extract of at least one food material selected from the following group:
dehydrogenase) inhibitor. a) Allspice, basil, coriander, nutmeg, pepper, sage, thyme, hop, glove (clove), cinnamon (cinnamon), mint (mint), licorice (licorice), fenka, rosemary, perilla, asenyaku, chrysanthemum And at least one spice or herb selected from the group consisting of gardenia. b) Coffee beans c) Cocoa beans d) Chlorella e) Mushrooms f) Rosaceae fruit g) Non-fermented tea h) Propolis 2. A hyaluronidase inhibitor having an excellent anti-inflammatory effect on hay fever, comprising a powder or extract of at least one food material selected from the following group: a) All spices, basil, coriander, sage,
At least one spice or herb selected from the group consisting of thyme, hops, gloves (cinch), cinnamon (cinnamon), mint (mint), licorice (licorice), fenca, rosemary, perilla, asenyaku, chrysanthemum, and gardenia. b) Coffee beans c) Cocoa beans d) Chlorella e) Mushrooms f) Rosaceae fruit g) Propolis