JPH0733325B2 - Cosmetics containing extract of cultured root of Cucurbitaceae plant - Google Patents

Description

DETAILED DESCRIPTION OF THE INVENTION [Industrial field of use] The present invention relates to a cosmetic composition containing an extract of a cultured root of a Cucurbitaceae plant. More specifically, a root induced by a plant hormone such as α-naphthalene acetic acid from a sprouting section of a Cucurbitaceae plant, or an octopine type Agrobacterium tumefaciens species (Agro
bacterium tumefaciens) (hereinafter abbreviated as A. tumefaciens)). A cosmetic composition comprising an extract of a cultured root obtained by culturing an induced root caused by infection.

[Conventional technology]

It has been conventionally known that a lotion is prepared by using a juice of fruits of a Cucurbitaceae plant or a juice leached from a stem. In particular, the juice leached from the loofah stem was used as loofah water as it was as lotion. However, the cultivation of Cucurbitaceae, which is an excellent base material for cosmetics, depends on the weather, so that there is a problem that a stable supply cannot be obtained by sampling from a cultivated product.

[Object of the Invention]

The object of the present invention is to cultivate cotyledons of the Cucurbitaceae plant, induced roots caused by plant hormones from tissue pieces such as hypocotyl, or induced roots caused by infection with octopine type A. tumefaciens. The purpose of the present invention is to add the obtained extract of cultured roots to cosmetics.

[Structure of Invention]

In order to achieve this object, the present invention, aseptic cultivated plants of the Cucurbitaceae, its cotyledons, from plant hormones such as hypocotyls, or aseptically grown seedlings hypocotyl and the like.
The method comprises causing induced roots by infecting A. tumefaciens, and adding the extract of the cultured roots obtained from these induced roots to cosmetics.

The Cucurbitacea used in the present invention is Cucurbitacea.
e), Cucumis melon (Cucumis melo)
L.) and Luffa cylindr
ica Roem.) is exemplified. Furthermore, as a melon,
Net melon [Cucumis melo L. (reticulatus grou
p)], Makuwauri [Cucumis melo L. (makuwagrou
p)], winter melon [Cucumis melo L. (inodorou
s group)], Shirouri (Cucumis melo L. (conomon gr
oup)], and a hybrid prince melon and the like.

In order to obtain a culture root of the melon of the genus Cucumber of the family Cucurbitaceae or the loofah of the genus Luffa of the family Cucurbitaceae, two methods, a method by addition of a plant hormone and a method by infection with A. tumefaciens, can be applied. . Hereinafter, a specific operation procedure will be described by taking melon as an example.

Melon seeds that have been sterilized with a sodium hypochlorite solution and washed with sterile water are germinated aseptically on agar, and cotyledons and hypocotyls 10 to 20 days after germination are cut with a sterilized scalpel into a suitable size. Murash containing auxins such as α-naphthalene acetic acid
Place and culture on agar such as ige-Skoog (MS) medium. By such culture, induced roots develop from the cut surface in the second week. The medium composition used to obtain the induced roots may be any medium such as MS medium, Linsmaier-Skoog (LS) medium, and White medium that are often used in normal plant tissue culture, but high concentrations such as MS medium Is preferred. An organic substance such as vitamins and amino acids, a carbon source, a plant hormone, and a natural extract are added to such a basic medium.

Examples of the organic substance include vitamins such as thiamine hydrochloride, nicotinic acid, pyridoxine and biotin, and amino acids such as glutamine, asparagine and alanine, but these trace organic substances do not necessarily need to be added to the medium.

Examples of the carbon source include sugars such as sucrose, grapes, maltose, and sorbitol, and the addition of sugars is preferable because the growth is quicker. Particularly, sucrose is preferably added, and 1 to 6% (W / V) is preferably added to the medium.

As a plant hormone, α-naphthalene acetic acid (NAA),
2,4-dichlorophenoxyacetic acid (2,4-D), 2,4,5-trichlorophenoxyacetic acid (2,4,5-T), indole-
Examples include auxins such as 3-acetic acid (IAA) and picloram (Pic), which are used alone or in combination. On the medium
It is preferable to add about 0.001 to 5 ppm.

As a natural extract, coconut milk (0-25% W /
V), casein hydrolyzate (0 to 2% W / V), yeast extract (0 to 2% W / V), and the like, and these can be used alone or in combination. In addition, in the culture in agar solid medium, in addition to the above medium composition components, agar (0.5 to 2% W
/ V) is added to solidify the medium.

The medium is finally adjusted to pH 5.4-6.0 with 0.1-1 N sodium hydroxide (or potassium hydroxide) and hydrochloric acid.

Culturing may be carried out in a bright or dark place under a constant condition of 20 to 35 ° C, preferably in a dark place around 28 ° C.

Next, a method for obtaining a root induced by infection with octopine type A. tumefaciens will be described in detail.

Examples of the octopine type A. tumefaciens used include known ones such as A6, B6 and A277 strains.

These A. tumefaciens are preferably cultivated prior to use. Usually, A. tumefaciens is inoculated into a medium such as LB (Luria-Bertani) medium, and the medium is incubated at 25-30 ° C for 24 hours.
After shaking culture for about 30 hours, a culture broth in a logarithmic growth phase can be obtained, which is used as it is or after static culture on an agar medium.

The A. tumefaciens used in the present invention is an octopine type A. tumefaciens, for example, A6, B.
Known ones such as 6, A277 strain can be used. These octopine type A. tumefaciens A6, B
6, A277 strain, etc., after obtaining permission to use the plant quarantine station of the Ministry of Agriculture, Forestry and Fisheries, fermentation research institute (Osaka, IFO13263, IFO1
3264, IFO13265), Department of Biology, Faculty of Science, Nagoya University, Dr. EWNester of Washington State University.

A. To infect cucurbitaceae plants with tumeficiens,
A method in which a stem of a plant individual is scratched and inoculated there is applied. In the case of melon, seeds that have been sterilized with an aqueous solution of sodium hypochlorite and washed with sterile water are germinated on agar, and the hypocotyls immediately below the cotyledons of the seedlings 3 to 5 days after germination are scratched, and A. Inoculate Mefaciens. Induced roots are formed 10 to 20 days after inoculation.

Cultured roots can be obtained by mass-producing the induced roots obtained in this manner by static culture, liquid culture (shaking culture, normal culture), etc. in the same manner as in the culture of general microorganisms. .

A known method, for example, a solvent extraction method, can be used to separate and collect the component excellent as the cosmetic base material from the culture root.

That is, the extract of the culture root used in the present invention, the culture root is, for example, water, ethanol, methanol, a water-soluble solvent such as propylene glycol, or, those extracted with a mixed solvent thereof, if necessary. It can be concentrated or diluted and used as a base material for cosmetics. The extraction operation is usually carried out by hot water extraction, preferably under pressure, for example, at a temperature of 120 to 200 ° C. for about 30 to 120 minutes. Further, an extraction method at a low temperature can also be used.
Although there is no particular limitation on the amount of the extraction solvent used, it is usually preferable to perform extraction using about 1 to 100 times the amount of the solvent based on the weight of the culture root. Further, the extraction can be repeated.

The obtained culture root extract can be used as a base, makeup, hair cosmetics, etc., together with various cosmetic base materials. Moreover, these cosmetics can be manufactured by a conventional method.

The extract of the culture root of the Cucurbitaceae plant used in the present invention is appropriately used in an amount of 0.001 to 20.0% (W / W) with respect to the cosmetic base material. If the content is less than 0.001% (W / W), the sufficient effect cannot be obtained, and if it exceeds 20.0% (W / W), the effect is not enhanced and it is uneconomical. The above-mentioned amount of the extract of the culture root of the Cucurbitaceae plant is used as a dried product (powder or extract) obtained by freeze-drying or concentrating the extract.
It is the compounding amount when is used. If the extract is used as it is, it is possible to add 100%.

〔The invention's effect〕

According to the present invention, a culture root of a Cucurbitaceae plant can be obtained by addition of a plant hormone or infection with an octopine type A. tumefaciens. According to this method, unlike agar cultivation, it is not affected by weather conditions,
It has the advantage that the plant tissue can be industrially planned and produced.

In addition, the present inventors previously found that the whole plant (leaf, stem,
It has been found that the root) and tissue culture callus extracts have excellent moisturizing properties and have an excellent skin-beautifying effect on the skin. Furthermore, when blended in a cosmetic, it has been found that it improves the color and luster of the skin, obtains an excellent skin conditioning effect such as prevention and improvement of fine wrinkles, and improves moisturizing properties. It was found that the same effect can be obtained with respect to the extract components of the cultured roots of the Cucurbitaceae plant obtained by the present invention.

Furthermore, regarding the water-extracted components of the cultured roots of the Cucurbitaceae plant of the present invention, a report by Hirai et al. (Biopharmaceutical Journal, 37, 374-380 (198
According to 3)), the histamine release inhibitory effect on mast cells collected from the rat abdominal cavity was measured. No histamine-releasing action was observed, and concanavalin A
(ConA) or compound 48/80 (Comp 48/80) was found to have an action of suppressing histamine release, and was found to be excellent as an anti-inflammatory agent (Table 1).

As is clear from Table 1, the water extract of the cultured roots of melon and loofah of the present invention showed the same anti-inflammatory action as the fruit of melon and loofah, or loofah water.

Hereinafter, the present invention will be described in more detail with reference to examples.

Example 1. Obtaining cultured roots from melon and loofah by using plant hormone Melon and loofah seeds were immersed in tap water overnight and then immersed in a 2.5% sodium hypochlorite solution for 10 to 15 minutes to adhere to the surface. After sterilizing the bacteria that are present, wash with sterile distilled water.
After absorbing the water with a paper towel, germinated aseptically on 0.8% (W / V) agar. Cultured in the light at 28 ℃ and after germination
When the size is about 5 mm, transfer to a 300 ml tall beaker containing 100 ml of MS medium and grow in a room with a white fluorescent lamp. On the 10th to 20th day of germination, the cotyledon of the seedling and the hypocotyl are 3m
MS medium (pH 5.7, pH 5.7, containing 3% sucrose, 0.1 ppm NAA, 0.8% agar)
Place in an autoclave at 120 ° C, 1.2 kg / cm ² ) for about 20 minutes and place it in a culture room (dark place) at 28 ° C for 2 weeks to obtain induced roots. In addition, the induced roots
The induced roots are grown by shaking culture in an MS liquid medium containing 3% sucrose and 0.1 ppm NAA. The melon and loofah culture roots thus grown are freeze-dried.

Example 2. Acquisition of cultured roots from melon and loofah by infection with octopine type A. tumefaciens.

Melon and loofah seeds were germinated aseptically in the same manner as in Example 1, the hypocotyl immediately below the cotyledon of the seedling after germination 3 to 5 days was damaged, and the A. tumefaciens A277 strain was inoculated therein. To do. As the A. tumefaciens A277 strain to be inoculated, one that has been shake-cultured in an LB (Luria-Bertani) medium at 30 ° C. for 24 to 30 hours is used. Induced roots are formed 10 to 20 days after inoculation.

Example 3. Water extract of melon and loofah culture roots 5.0 g of melon and loofah culture root (lyophilized product) were crushed and then extracted with 300 ml of water at 95 ° C for 3 hours. After heating, the residue was separated, and then the liquid was freeze-dried to obtain 1.5 g and 1.7 g of pale white to pale yellow powders, respectively.

Example 4. Lotion formulation amount A) Water extract of melon culture root 0.7 parts 1,3-butylene glycol 8.0 gericerin 2.0 xanthan gum 0.2 purified water 44.6 B) ethanol 5.0 preservative proper amount polyoxyethylene (40) hydrogenated castor oil 0.1 Perfume Proper amount Purified water 10.0 Manufacturing method: Components A and B are uniformly dissolved and then mixed to obtain a product.

Example 5 Lotion Toner lotion was obtained in the same manner as in Example 4, except that the water extract of cultivated loofah was used instead of the water extract of melon-cultured root.

Example 6. Skin cream formulation Amount: A) stearic acid 4.0 parts cetyl alcohol 3.0 stearyl alcohol 1.0 liquid paraffin 6.5 vaseline 10.0 sorbitan monostearate 1.5 polyoxyethylene (25) monostearate 3.0 B) 1,3-butylene glycol 5.0 Potassium hydroxide 0.1 Water extract of melon culture root 0.8 Preservative Suitable amount Purified water 65.1 C) Perfume Suitable amount Production method: Oil phase component A and water phase component B are 70-
After heat-dissolving at 75 ° C., component B is added to component A to emulsify, component C is added and mixed during cooling, and cooled to 30 ° C. to obtain a product.

Example 7. Emulsion formulation Formulation amount A) Stearic acid 5.0 parts Cetyl alcohol 5.0 Liquid paraffin 2.0 Glycerin monostearate 1.3 Sorbitan monooleate 1.5 Polyoxyethylene (10) sorbitan monooleate 0.8 B) Glycerin 6.0 Water extract of loofah culture root 0.5 Preservative Suitable amount Purified water 77.9 C) Fragrance Suitable amount Manufacturing method: A product is prepared in the same manner as in Example 6.

Example 8. Pack prescription compounding amount glycerin 15.0 1,3-butylene glycol 10.0 polyoxyethylene (40) hydrogenated castor oil 0.5 water extract of melon culture root 2.0 citric acid 0.1 sodium citrate 0.3 preservative proper amount perfume proper purified water 72.1 Manufacturing method: Dissolve each component uniformly to obtain a product.

In order to clarify the safety of the cultured roots of the Cucurbitaceae plant of the present invention, a primary irritation test on a human body was conducted by an occlusive patch test. That is, using a fin chamber (Taisho Pharmaceutical Co., Ltd.), 30 healthy people were occluded on the flexed side of the forearm for 48 hours, and after 1 hour, 24 hours, and 48 hours after removal of the patch test plaster. It judged using the average value of the judgment of. No clear erythema was observed in any of the extracts of the cultured roots of the Cucurbitaceae plant, confirming low primary irritation. The test was conducted using 10% (W / W) aqueous solutions as the samples (Table 2).

Further, in order to clarify the effect of the cosmetic composition of the present invention, a use test was conducted on 30 general women aged 25 to 50, and a questionnaire survey was conducted mainly on the skin conditioning effect by the double blind method. Lotion containing water extract component of melon culture root (Example 4), lotion containing water extract component of loofah culture root (Example 5), and conventional makeup containing no water extract of melon or loofah culture root at all. The results of using water (Example 1) for one month are summarized in Table 5. No skin trouble occurred during the period of use.

As is clear from Table 3, the addition of the melon or the extract of cultured loofah roots provided an excellent skin conditioning effect.

Claims (1)

[Claims] 1. A cosmetic comprising an extract of a cultured root obtained from a root induced by tissue culture of a Cucurbitaceae plant or infection with an octopine type Agrobacterium tumefaciens. .