JP7466884B2 - Type I collagen production promoter in normal human gingival fibroblasts - Google Patents

Description

The present invention relates to a type I collagen production promoter in normal human gingival fibroblasts and an oral preparation.

Gums are one of the periodontal tissues that support the teeth and surround the roots of the teeth. They consist of two layers: the epithelial tissue that contacts the outside world and the dermal tissue underneath the epithelial tissue. It is said that about 60% of the gums is collagen, which is responsible for the gums' elasticity and adhesion.

Gingival fibroblasts are present in the dermis tissue, and are known to produce type I collagen. It is also known that the production of type I collagen in gingival fibroblasts decreases with age and due to toxins released by bacteria causing periodontal disease.
Therefore, in order to improve the health of periodontal tissues, it is thought that promoting the production of type I collagen in gingival fibroblasts is necessary.

To date, compositions have been proposed for medicinal use to maintain and improve gums, including compositions that contain astaxanthin and/or fatty acid esters of astaxanthin as active ingredients and that suppress and prevent apoptosis of gingival cells (see, for example, Patent Document 1).
However, the above proposal does not promote the production of type I collagen in gingival fibroblasts.

Therefore, there is currently a strong demand for the rapid development of a highly safe substance that has excellent type I collagen production promoting effects in normal human gingival fibroblasts.

JP 2015-110526 A

The present invention aims to solve the above-mentioned conventional problems and to achieve the following object: That is, the present invention aims to provide a type I collagen production promoter in normal human gingival fibroblasts that has an excellent type I collagen production promoting effect in normal human gingival fibroblasts and is highly safe.
Another object of the present invention is to provide an oral preparation which has an excellent type I collagen production promoting effect in normal human gingival fibroblasts and is highly safe.

As a result of intensive research conducted by the present inventors to solve the above problems, it was discovered that at least one selected from the group consisting of dipotassium glycyrrhizinate, wolfberry extract, Scutellaria root extract, licorice extract, carrot extract, royal jelly extract, Hypericum perforatum extract, Sophora japonica extract, and sweet tea extract has an excellent effect of promoting type I collagen production in normal human gingival fibroblasts, and the present invention was completed.

The present invention is based on the above-mentioned findings of the present inventors, and the means for solving the above-mentioned problems are as follows.
<1> A type I collagen production promoter in normal human gingival fibroblasts, comprising at least one selected from the group consisting of dipotassium glycyrrhizinate, wolfberry extract, Scutellaria root extract, licorice extract, carrot extract, royal jelly extract, Hypericum perforatum extract, Sophora japonica extract, and sweet tea extract.
<2> An oral preparation, comprising the promoter for type I collagen production in normal human gingival fibroblasts according to <1>.

The type I collagen production promoter in normal human gingival fibroblasts of the present invention solves the above-mentioned conventional problems, and provides a type I collagen production promoter in normal human gingival fibroblasts that has excellent type I collagen production promoting effect in normal human gingival fibroblasts and is highly safe.
The oral preparation of the present invention solves the above-mentioned conventional problems, and provides an oral preparation that has an excellent type I collagen production promoting effect in normal human gingival fibroblasts and is highly safe.

(Type I collagen production promoter in normal human gingival fibroblasts)
The type I collagen production promoter of the present invention in normal human gingival fibroblasts contains at least one selected from the group consisting of dipotassium glycyrrhizinate, wolfberry extract, Scutellaria root extract, licorice extract, carrot extract, royal jelly extract, Hypericum perforatum extract, Sophora japonica extract and sweet tea extract, and further contains other ingredients as necessary.

The fact that the above dipotassium glycyrrhizinate, wolfberry extract, Scutellaria root extract, licorice extract, honeysuckle extract, royal jelly extract, Hypericum perforatum extract, Sophora japonica extract and sweet tea extract have excellent type I collagen production promoting effects in normal human gingival fibroblasts and are useful as type I collagen production promoters in normal human gingival fibroblasts was not previously known, and is a new discovery by the present inventors.

The dipotassium glycyrrhizinate, wolfberry extract, Scutellaria root extract, licorice extract, honeysuckle extract, royal jelly extract, Hypericum perforatum extract, Sophora japonica extract, and sweet tea extract may be used alone or in combination of two or more.

<Dipotassium glycyrrhizinate>
Dipotassium glycyrrhizinate is a compound represented by C ₄₂ H ₆₀ K ₂ O ₁₆ and has a molecular weight of 899.1.
Dipotassium glycyrrhizinate used may be prepared by chemical synthesis or may be a commercially available product.
The method of chemical synthesis is not particularly limited and can be appropriately selected depending on the purpose. For example, there is a method of reacting glycyrrhizinic acid with potassium hydroxide (KOH) or the like.

The method for obtaining the glycyrrhizinic acid is not particularly limited and can be appropriately selected depending on the purpose. For example, a method using a commercially available product, a method for obtaining the glycyrrhizinic acid by at least one of isolation and purification methods from a plant extract containing glycyrrhizinic acid, etc. can be mentioned.

The plant extract containing glycyrrhizinic acid may be an extract obtained using a plant containing glycyrrhizinic acid as an extraction raw material, a diluted or concentrated solution of the extract, a dried product obtained by drying the extract, or a crude or purified product thereof.
The plant extract containing glycyrrhizic acid can be obtained by a method generally used for extracting plants. An example of a plant containing glycyrrhizic acid is licorice.

<Wolfberry extract>
Wolfberry (scientific name: Lycium chinense or Lycium barbarum ) is a plant belonging to the genus Lycium in the family Solanaceae, and is widely distributed in East Asia and can be easily obtained in these areas.

The wolfberry extract may be prepared from the part of the wolfberry used as the extraction raw material, or a commercially available product may be used.
The part of the wolfberry to be extracted is not particularly limited and can be appropriately selected depending on the purpose, but the leaves are preferred.
The method for preparing the wolfberry extract is not particularly limited and can be appropriately selected depending on the purpose. For example, the wolfberry extract can be prepared by the method for preparing extracts of natural products described below.

<Scutellaria root extract>
Scutellaria baicalensis (herbal medicine name) is the root of Scutellaria baicalensis (scientific name: Scutellaria baicalensis Georgi), a perennial plant belonging to the genus Scutellaria of the family Lamiaceae, distributed in northern and northeastern China, Mongolia, etc., and can be easily obtained from these areas. Scutellaria baicalensis has traditionally been used as a stomachic, antiallergic agent, etc.

The Scutellaria root extract may be prepared from the roots of Scutellaria baicalensis or a commercially available product.
The method for preparing the Scutellaria root extract is not particularly limited and may be appropriately selected depending on the purpose. For example, the extract may be prepared by the method for preparing extracts of natural products described below.

<Licorice extract>
Licorice (Glycyrrhiza) is a perennial herb belonging to the genus Glycyrrhiza of the family Fabaceae, and has been used since ancient times for medicinal purposes and as a raw material for sweeteners. The licorice includes Glychyrrhiza glabra , Glychyrrhiza inflata , Glychyrrhiza uralensis, Glychyrrhiza aspera , Glychyrrhiza eurycarpa , Glychyrrhiza pallidiflora, Glychyrrhiza yunnanensis , Glychyrrhiza lepidota, Glychyrrhiza glabra , Glychyrrhiza inflata, Glychyrrhiza uralensis , Glychyrrhiza lepidota ... There are various types, such as Glychyrrhiza lepidota , Glychyrrhiza echinata , and Glychyrrhiza acanthocarpa .

The licorice extract may be prepared from the part of the licorice plant used as the raw material for extraction, or a commercially available product may be used.
The type of licorice used as the extraction raw material is not particularly limited and can be appropriately selected depending on the purpose.
The part of licorice to be extracted is not particularly limited and may be appropriately selected depending on the purpose, and examples thereof include leaves, branches, bark, trunks, stems, fruits, seeds, flowers and other above-ground parts, roots, rhizomes, and mixtures of these parts. Among these, roots, leaves, and rhizomes are preferred.
The method for preparing the licorice extract is not particularly limited and can be appropriately selected depending on the purpose. For example, the licorice extract can be prepared by a method for preparing extracts of natural products, which will be described later.

<Honeybee Extract>
Lonicera japonica (scientific name: Lonicera japonica ) is a plant belonging to the Caprifoliaceae family and grows in various parts of China, Korea, and Japan, and can be easily obtained from these areas.

The above-mentioned ginseng extract may be prepared from the part of the plant used as the raw material for extraction, or a commercially available product may be used.
The part of the plant from which the extract is made is not particularly limited and can be appropriately selected depending on the purpose, and examples thereof include flower buds, leaves, branches, seeds, bark, roots, stems, or mixtures of these parts. Among these, leaves and stems are preferred.
The method for preparing the ginseng extract is not particularly limited and can be appropriately selected depending on the purpose. For example, the extract can be prepared by a method for preparing an extract of a natural product, which will be described later.

<Royal jelly extract>
Royal jelly is a secretion from the pharyngeal glands of young worker bees of the European honeybee (scientific name: Apis melifera L., also known as the Western honeybee) or the Asian honeybee (scientific name: Apis indeca Radoszkowski), which belongs to the genus Apis and the family Apidae and are distributed in Europe, Africa, etc., and is fed to the larvae that will become queen bees and to the queen bees that have become adults.

The royal jelly extract to be used may be one prepared from royal jelly, or a commercially available product may be used.
The method for preparing the royal jelly extract is not particularly limited and can be appropriately selected depending on the purpose. For example, the royal jelly extract can be prepared by the method for preparing extracts of natural products described below.

<Hypericum extract>
St. John's wort ( Hypericum erectum , Hypericum perforatum ) is a perennial herb belonging to the genus Hypericum in the family Hypericaceae, which grows naturally in Honshu, Shikoku, Kyushu, etc. and can be easily obtained from these areas.

The Hypericum extract may be prepared from the part of the plant used as the raw material for extraction, or a commercially available product may be used.
The part of Hypericum to be extracted is not particularly limited and may be appropriately selected depending on the purpose, and examples thereof include leaves, stems, roots, flowers, above-ground parts, and mixtures of these parts. Among these, above-ground parts and the whole plant are preferred.
The method for preparing the Hypericum extract is not particularly limited and can be appropriately selected depending on the purpose. For example, the extract can be prepared by a method for preparing an extract of a natural product, which will be described later.

<Moroccanus arbutus extract>
Morus alba (herbal medicine name) is the root bark of Morus alba (scientific name: Morus alba L.), a deciduous tall tree of the genus Morus in the family Mulberry, which is distributed in China and other areas, and can be easily obtained from these areas. Morus alba has traditionally been used as an anti-inflammatory, diuretic, antipyretic, analgesic, etc.

The above-mentioned Morus alba extract may be prepared from the root bark of Morus alba or may be a commercially available product.
The method for preparing the Morus albicans extract is not particularly limited and may be appropriately selected depending on the purpose. For example, the extract may be prepared by a method for preparing an extract of a natural product, which will be described later.

<Tencha extract>
Rubus suavissimus is a shrub belonging to the genus Rubus in the family Rosaceae, which grows naturally in southern China and can be easily obtained from these regions. Tencha extracts have been used as antiallergic agents and the like.

The sweet tea extract may be prepared from the part of the tea plant used as the raw material for extraction, or a commercially available product may be used.
The part of the sweet tea to be extracted is not particularly limited and can be appropriately selected depending on the purpose, and examples thereof include above-ground parts such as leaves, stems, flowers, and fruits, underground parts such as roots, and mixtures of these parts. Among these, leaves are preferred.
The method for preparing the sweet tea extract is not particularly limited and can be appropriately selected depending on the purpose. For example, the extract can be prepared by a method for preparing an extract of a natural product, which will be described later.

-Method for preparing extracts of natural products-
The above-mentioned wolfberry extract, Scutellaria root extract, licorice extract, honeysuckle extract, royal jelly extract, Hypericum extract, Sophora japonica extract and sweet tea extract (hereinafter sometimes referred to as "extracts of various natural products") can be easily obtained by utilizing a method generally used for plant extraction. The extracts of various natural products include extracts of various natural products, dilutions or concentrates of the extracts, dried products of the extracts, or crude or purified products thereof.

The method for preparing the extracts from the various natural products is not particularly limited and can be appropriately selected depending on the purpose. For example, the extracts can be obtained by drying the raw material for extraction, and then subjecting it to extraction with an extraction solvent, either directly or after crushing using a crusher. The drying can be performed in the sun or using a commonly used dryer.
In addition, the extract may be used after pretreatment such as degreasing with a non-polar solvent such as hexane. By carrying out pretreatment such as degreasing, extraction with a polar solvent can be carried out efficiently.

The extraction solvent is not particularly limited and can be appropriately selected depending on the purpose. Examples of the extraction solvent include water, a hydrophilic organic solvent, or a mixture thereof.

There are no particular limitations on the water that can be used as the extraction solvent, and it can be appropriately selected depending on the purpose. Examples of the water include pure water, tap water, well water, mineral water, hot spring water, spring water, fresh water, and water that has been subjected to various treatments. Treatments that can be applied to water include, for example, purification, heating, sterilization, filtration, ion exchange, adjustment of osmotic pressure, buffering, and the like. Therefore, water that can be used as the extraction solvent also includes purified water, hot water, ion-exchanged water, physiological saline, phosphate buffer, phosphate buffered physiological saline, and the like.

There are no particular limitations on the hydrophilic organic solvent that can be used as the extraction solvent for the plant, and it can be appropriately selected depending on the purpose. For example, it can be lower alcohols having 1 to 5 carbon atoms, such as methanol, ethanol, propyl alcohol, isopropyl alcohol, etc.; lower aliphatic ketones, such as acetone, methyl ethyl ketone, etc.; polyhydric alcohols having 2 to 5 carbon atoms, such as 1,3-butylene glycol, propylene glycol, glycerin, etc.; and mixed solvents of these hydrophilic organic solvents with water can also be used.

When using a mixed solvent of water and the hydrophilic organic solvent, the mixing ratio is not particularly limited and can be appropriately selected depending on the purpose. For example, in the case of a mixed solvent of water and a lower alcohol, the mixing ratio of water to lower alcohol is preferably 9:1 to 1:9 (volume ratio), and more preferably 8:2 to 2:8 (volume ratio). In addition, in the case of a mixed solvent of water and a lower aliphatic ketone, the mixing ratio of water to lower aliphatic ketone is preferably 9:1 to 2:8 (volume ratio). In addition, in the case of a mixed solvent of water and a polyhydric alcohol, the mixing ratio of water to polyhydric alcohol is preferably 5:5 to 1:9 (volume ratio).

The extraction solvent is preferably used at room temperature or at a temperature below the boiling point of the solvent.

The extraction method is not particularly limited and can be appropriately selected depending on the purpose. For example, the extraction raw material is immersed in an extraction solvent having an amount (mass ratio) of 5 to 15 times the amount of the extraction raw material, and soluble components are extracted at room temperature or under reflux heating, followed by filtration to remove the extraction residue to obtain an extract. Next, the solvent is distilled off from the extract to obtain a paste-like concentrate, and this concentrate is further dried to obtain a dried product.
The conditions for the extraction are not particularly limited and can be appropriately selected depending on the purpose.

In addition, the obtained extracts of the various natural products may be subjected to treatments such as dilution, concentration, drying, purification, etc. in accordance with conventional methods in order to obtain diluted products, concentrates, dried products, crude products, purified products, etc. of the extracts of the various natural products.
The above-obtained extracts of various natural products can be used as they are as type I collagen production promoters in normal human gingival fibroblasts, but the above-mentioned concentrated solutions and dried products are preferred in terms of ease of use. In obtaining the dried products, carriers such as dextrin and cyclodextrin may be added to improve hygroscopicity.

The content of at least one type of promoter of type I collagen production in normal human gingival fibroblasts selected from the group consisting of dipotassium glycyrrhizinate, wolfberry extract, Scutellaria root extract, licorice extract, carrot extract, royal jelly extract, Hypericum perforatum extract, Sophora japonica extract, and sweet tea extract is not particularly limited and can be appropriately selected depending on the purpose.

<Other ingredients>
The other components are not particularly limited and can be appropriately selected depending on the purpose. Examples of the other components include excipients, moisture-proofing agents, preservatives, strengthening agents, thickeners, emulsifiers, antioxidants, sweeteners, acidulants, seasonings, colorants, fragrances, whitening agents, moisturizers, oily components, UV absorbers, surfactants, alcohols, powder components, colorants, aqueous components, water, and skin nutrients.
The content of the other components in the promoter for type I collagen production in normal human gingival fibroblasts is not particularly limited and can be appropriately selected depending on the purpose.

The above dipotassium glycyrrhizinate, wolfberry extract, Scutellaria root extract, licorice extract, honeysuckle extract, royal jelly extract, Hypericum perforatum extract, Sophora japonica extract and sweet tea extract have excellent type I collagen production promoting effects in normal human gingival fibroblasts, and therefore can be used as active ingredients of type I collagen production promoters in normal human gingival fibroblasts.

<Applications>
The use of the type I collagen production promoter in normal human gingival fibroblasts of the present invention is not particularly limited and can be appropriately selected depending on the purpose. For example, it can be suitably used as an oral preparation as described below, and its content, method of use and dosage form can be appropriately selected depending on the purpose of use.

(Oral preparations)
The oral preparation of the present invention contains the promoter of type I collagen production in normal human gingival fibroblasts of the present invention, and further contains other ingredients as required.

The content of the type I collagen production promoter in normal human gingival fibroblasts in the oral preparation is not particularly limited and can be appropriately selected depending on the purpose. The oral preparation may consist only of the type I collagen production promoter in normal human gingival fibroblasts.

The other components in the oral preparation are not particularly limited and can be appropriately selected depending on the purpose. For example, main components, auxiliary components or other components used in the production of ordinary oral preparations can be used in combination. More specific examples include abrasives such as calcium hydrogen phosphate, calcium carbonate, insoluble sodium metaphosphate, aluminosilicate, anhydrous silicic acid, and resin; surfactants such as long-chain sodium alkyl sulfate, sodium lauryl sulfoacetate, lauryl diethanolamide, and sucrose fatty acid esters; binders such as carboxymethylcellulose (CMC), hydroxyethylcellulose, alginates, carrageenan, gum arabic, and polyvinyl alcohol; thickeners such as polyethylene glycol, sorbitol, glycerin, and propylene glycol; sweeteners such as saccharin, steviosides, glycyrrhizic acid, and thaumatin; preservatives such as dehydroacetic acid and sodium dehydroacetate; fragrances such as menthol, carvone, eugenol, anethole, peppermint oil, spearmint oil, peppermint oil, eucalyptus oil, ginger oil, and anise oil; and various pigments. These may be used alone or in combination of two or more.
The content of other ingredients in the oral preparation is not particularly limited and can be appropriately selected depending on the purpose.

The oral preparation of the present invention has excellent type I collagen production promotion properties in normal human gingival fibroblasts, and therefore can be suitably used, for example, for promoting collagen regeneration in periodontal tissues.

The form of the oral preparation is not particularly limited and can be appropriately selected depending on the purpose. Examples include toothpaste, mouthwash, lozenges, gum massage cream, gargle, mouth freshener, etc.

The type I collagen production promoter in normal human gingival fibroblasts and the oral preparation of the present invention are preferably applied to humans, but can also be applied to animals other than humans as long as their respective functional effects are achieved.

The following describes manufacturing and test examples of the present invention, but the present invention is not limited to these manufacturing and test examples.

The materials used in Test Examples 1-1 and 1-2 described below are as follows.
(1) Dipotassium glycyrrhizinate (External Product Dipotassium Glycyrrhizinate, manufactured by Maruzen Pharmaceutical Co., Ltd.)
(2) Scutellaria root extract (Scutellaria root extract BG-LC (extraction raw material: Scutellaria baicalensis (root), extraction solvent: 50% by volume 1,3-butylene glycol), manufactured by Maruzen Pharmaceutical Co., Ltd.)
(3) Licorice extract (Licorice extract (extraction raw material: licorice (root), extraction solvent: water), manufactured by Maruzen Pharmaceutical Co., Ltd.)
(4) Panax ginseng extract (Panax ginseng extract-J (extraction raw material: Panax ginseng (leaves), extraction solvent: 90% by volume ethanol), manufactured by Maruzen Pharmaceutical Co., Ltd.)
(5) Royal jelly extract (Royal jelly extract BG (extraction solvent: 50% by volume 1,3-butylene glycol), manufactured by Maruzen Pharmaceutical Co., Ltd.)
(6) Hypericum extract (Hypericum extract-J (extraction raw material: Hypericum perforatum (flowers, leaves and stems), extraction solvent: 90% by volume ethanol), manufactured by Maruzen Pharmaceutical Co., Ltd.)
(7) Morus alba extract (Morus alba extract BG (extraction raw material: Morus alba (root bark), extraction solvent: 50% by volume 1,3-butylene glycol), manufactured by Maruzen Pharmaceutical Co., Ltd.)
(8) Sweet Tea Extract (Tencha Extract BGW (extraction raw material: sweet tea (leaves), extraction solvent: 50% by volume 1,3-butylene glycol), manufactured by Maruzen Pharmaceutical Co., Ltd.)
(9) Wolfberry extract (see Production Examples 1 to 3 below)

(Production Examples 1 to 3)
-Production of wolfberry extract-
20 g of dried wolfberry leaves were added with 10 times the amount of solvent (see Table 1 below) and refluxed for 2 hours. After filtration under reduced pressure, the solvent was removed to obtain each extract. The yields are also shown in Table 1.

(Test Example 1-1)
<Test on promoting type I collagen production in NHGF cells>
Each of the above-mentioned materials was used as a test sample, and the promoting effect on type I collagen production in NHGF cells was tested by the following test method.

Normal human gingival fibroblasts were cultured in a low serum medium for normal human fibroblast proliferation (LFK-LS1038), and then the cells were harvested by trypsinization. The harvested cells were diluted with LFK-LS1038 medium to a concentration of 1.6 x ¹⁰⁵ cells/mL, and then seeded in a 96-well plate at 100 μL per well and cultured for 1 day.
After completion of the culture, 100 μL of the test sample (for sample concentration, see Tables 2-1 and 2-2 below) dissolved in serum-free medium for growing normal human fibroblasts (LFK-LS1010) was added to each well and cultured for 3 days. As a control, culture was performed in the same manner using LFK-LS1010 medium without adding any sample. After completion of the culture, the amount of type I collagen in the medium in each well was measured by ELISA. From the measurement results, the type I collagen production promotion rate (%) was calculated using the following formula 1.
The results are shown in the upper rows of Tables 2-1 and 2-2.
<Formula 1>
Type I collagen production promotion rate (%) = A/B x 100
In formula 1, "A" represents the "amount of type I collagen when a test sample is added," and "B" represents the "amount of type I collagen when no sample is added."

(Test Example 1-2)
<Test on promoting type I collagen production in NB1RGB cells>
Each of the above-mentioned materials was used as a test sample, and the type I collagen production promoting effect in NB1RGB cells was tested by the following test method.

Normal human skin fibroblasts (NB1RGB cells) were cultured in Dulbecco's MEM medium containing 10% FBS, and then the cells were harvested by trypsinization. The harvested cells were diluted with the above medium to a concentration of 1.6 x ¹⁰⁵ cells/mL, and then seeded in a 96-well plate at 100 μL per well and cultured overnight.
After completion of the culture, the medium was removed, and 150 μL of the test sample dissolved in 0.25% FBS-containing Dulbecco's MEM medium (see Tables 2-1 and 2-2 below for sample concentration) was added to each well and cultured for 3 days. As a control, culture was performed in the same manner using 0.25% FBS-containing Dulbecco's MEM medium without the addition of sample. After completion of the culture, the amount of type I collagen in the medium in each well was measured by ELISA. From the measurement results, the type I collagen production promotion rate (%) was calculated using the following formula 2.
The results are shown in the lower part of Tables 2-1 and 2-2.
<Formula 2>
Type I collagen production promotion rate (%) = C/D x 100
In formula 2, "C" represents the "amount of type I collagen when a test sample is added," and "D" represents the "amount of type I collagen when no sample is added."

From the results in Tables 2-1 and 2-2, in normal human skin fibroblasts, the promotion rate of type I collagen production was at most about 114%.
On the other hand, when the same type and concentration of material was used, the rate of promotion of type I collagen production in normal human gingival fibroblasts was at least higher than the rate of promotion of type I collagen production in normal human skin fibroblasts.
Therefore, it was confirmed that dipotassium glycyrrhizinate, wolfberry extract, Scutellaria root extract, licorice extract, honeysuckle extract, royal jelly extract, Hypericum perforatum extract, Sophora japonica extract and sweet tea extract exhibit excellent type I collagen production promoting effects specifically in normal human gingival fibroblasts, and these materials are considered to be excellent type I collagen production promoters in normal human gingival fibroblasts and active ingredients for oral preparations.

(Formulation Example 1)
A mouthwash having the following composition was prepared by a conventional method.
Dipotassium glycyrrhizinate 0.2 parts by mass (external glycyrrhizinate dipotassium, manufactured by Maruzen Pharmaceutical Co., Ltd.)
Dicalcium phosphate 45.0 parts by weight Sodium carboxymethylcellulose 1.0 part by weight Glycerin 20.0 parts by weight Sodium lauryl sulfate 2.0 parts by weight Fragrance 1.0 part by weight Purified water 30.0 parts by weight

(Formulation Example 2)
A mouthwash having the following composition was prepared by a conventional method.
0.2 parts by weight of Scutellaria root extract (Scutellaria root extract BG-LC, manufactured by Maruzen Pharmaceutical Co., Ltd.)
Dicalcium phosphate 45.0 parts by weight Sodium carboxymethylcellulose 1.0 part by weight Glycerin 20.0 parts by weight Sodium lauryl sulfate 2.0 parts by weight Fragrance 1.0 part by weight Purified water 30.0 parts by weight

(Formulation Example 3)
A mouthwash having the following composition was prepared by a conventional method.
Licorice extract: 0.2 parts by mass (Licorice extract, manufactured by Maruzen Pharmaceutical Co., Ltd.)
Dicalcium phosphate 45.0 parts by weight Sodium carboxymethylcellulose 1.0 part by weight Glycerin 20.0 parts by weight Sodium lauryl sulfate 2.0 parts by weight Fragrance 1.0 part by weight Purified water 30.0 parts by weight

(Formulation Example 4)
A mouthwash having the following composition was prepared by a conventional method.
0.2 parts by mass of ginseng extract (ginseng extract-J, manufactured by Maruzen Pharmaceutical Co., Ltd.)
Dicalcium phosphate 45.0 parts by weight Sodium carboxymethylcellulose 1.0 part by weight Glycerin 20.0 parts by weight Sodium lauryl sulfate 2.0 parts by weight Fragrance 1.0 part by weight Purified water 30.0 parts by weight

(Formulation Example 5)
A toothpaste having the following composition was prepared in a conventional manner.
Royal jelly extract: 0.1 parts by mass (Royal jelly extract BG, manufactured by Maruzen Pharmaceutical Co., Ltd.)
Calcium carbonate 39.0 parts by weight Sorbitol 22.0 parts by weight Carboxymethylcellulose 1.1 parts by weight Sodium lauryl sulfate 1.3 parts by weight Saccharin 0.1 parts by weight Fragrance 1.0 part by weight Chlorhexidine hydrochloride 0.01 parts by weight Purified water 35.0 parts by weight

(Formulation Example 6)
A toothpaste having the following composition was prepared in a conventional manner.
Hypericum extract: 0.1 parts by mass (Hypericum extract-J, manufactured by Maruzen Pharmaceutical Co., Ltd.)
Calcium carbonate 39.0 parts by weight Sorbitol 22.0 parts by weight Carboxymethylcellulose 1.1 parts by weight Sodium lauryl sulfate 1.3 parts by weight Saccharin 0.1 parts by weight Fragrance 1.0 part by weight Chlorhexidine hydrochloride 0.01 parts by weight Purified water 35.0 parts by weight

(Formulation Example 7)
A toothpaste having the following composition was prepared in a conventional manner.
- 0.1 parts by mass of mulberry bark extract (mulberry bark extract BG, manufactured by Maruzen Pharmaceutical Co., Ltd.)
Calcium carbonate 39.0 parts by weight Sorbitol 22.0 parts by weight Carboxymethylcellulose 1.1 parts by weight Sodium lauryl sulfate 1.3 parts by weight Saccharin 0.1 parts by weight Fragrance 1.0 part by weight Chlorhexidine hydrochloride 0.01 parts by weight Purified water 35.0 parts by weight

(Formulation Example 8)
A toothpaste having the following composition was prepared in a conventional manner.
0.1 parts by mass of sweet tea extract (sweet tea extract BGW, manufactured by Maruzen Pharmaceutical Co., Ltd.)
Calcium carbonate 39.0 parts by weight Sorbitol 22.0 parts by weight Carboxymethylcellulose 1.1 parts by weight Sodium lauryl sulfate 1.3 parts by weight Saccharin 0.1 parts by weight Fragrance 1.0 part by weight Chlorhexidine hydrochloride 0.01 parts by weight Purified water 35.0 parts by weight

(Formulation Example 9)
A mouthwash having the following composition was prepared by a conventional method.
Wolfberry extract: 0.2 parts by mass (wolfberry leaf hot water extract of Production Example 1)
Dicalcium phosphate 45.0 parts by weight Sodium carboxymethylcellulose 1.0 part by weight Glycerin 20.0 parts by weight Sodium lauryl sulfate 2.0 parts by weight Fragrance 1.0 part by weight Purified water 30.0 parts by weight

(Formulation Example 10)
A toothpaste having the following composition was prepared in a conventional manner.
Wolfberry extract: 0.1 parts by mass (50% by volume ethanol extract of wolfberry leaves of Production Example 2)
Calcium carbonate 39.0 parts by weight Sorbitol 22.0 parts by weight Carboxymethylcellulose 1.1 parts by weight Sodium lauryl sulfate 1.3 parts by weight Saccharin 0.1 parts by weight Fragrance 1.0 part by weight Chlorhexidine hydrochloride 0.01 parts by weight Purified water 35.0 parts by weight

(Formulation Example 11)
A toothpaste having the following composition was prepared in a conventional manner.
Wolfberry extract: 0.1 parts by mass (wolfberry leaf 80% ethanol extract of Production Example 3)
Calcium carbonate 39.0 parts by weight Sorbitol 22.0 parts by weight Carboxymethylcellulose 1.1 parts by weight Sodium lauryl sulfate 1.3 parts by weight Saccharin 0.1 parts by weight Fragrance 1.0 part by weight Chlorhexidine hydrochloride 0.01 parts by weight Purified water 35.0 parts by weight

Claims (1)

A type I collagen production promoter in normal human gingival fibroblasts, characterized by containing at least one selected from dipotassium glycyrrhizinate and licorice extract.