JP6125864B2 - Dendritic negative regulator - Google Patents

Description

  The present invention relates to a dendrite negative control agent and a screening method for a dendrite control agent.

Dendrites are a plurality of protrusions that branch from a cell body like a tree branch in order to receive information sent from an external stimulus or an axon of another nerve cell.
In recent years, the role of dendrites of cells other than nerve cells has attracted attention. For example, melanosomes containing mature melanin granules migrate within cells and are passed from melanocyte dendrites to adjacent keratinocytes. At this time, formation of dendrites of melanocytes is considered to be important, but the mechanism of dendrite formation has not been fully elucidated.
For this reason, research on the control of dendrites has been further advanced.
For example, Patent Document 1 proposes to provide a technique for suppressing Rac activity and contracting dendrites to the same level in order to screen for Rac activity promoters useful for nerve cell regeneration and the like. ing.
Further, for example, Patent Document 2 proposes a melanocyte dendrite elongation inhibitor composed of the essence of Citrus agonia.
Moreover, for example, Patent Document 3 proposes a melanocyte dendrite formation inhibitor containing yeast extract as an active ingredient.
However, it is still unclear what process dendrite formation is formed through, and further research and development of substances capable of controlling dendrite formation is required.

JP 2006-067940 A JP 2001-199866 A JP 2003-252742 A

  Therefore, in view of such circumstances, the present invention intends to provide an excellent dendritic negative control agent and a screening method for a dendritic positive control agent using the same.

Thus, as a result of intensive studies, the present inventors have found that an extract of Rosaceae Karin, which is considered to be highly safe due to natural origin, negatively controls dendrites, and completed the present invention.
That is, this invention provides the dendrite negative control inhibitor which uses the extract of a Rosaceae karin as an active ingredient.
The plant may be an extract from the fruit of the plant.
The scientific name of the plant may be any of Chaenomeles sinensis (Thouin) Koehne, Chaenomeles sinensis, Pseudocydonia sinensis, and Cydonia sinensis Thouin.
It is preferable that the plant extract is selected from water, alcohols, and hydrous alcohols. The alcohols preferably have 1 to 4 carbon atoms. The alcohol may be selected from methanol, ethanol, and 1,3-butanediol. The water concentration may be 50 to 100 vol%.
The dendrite negative control may be suppression of dendrite formation, regression of dendrite, or suppression of dendrite activation.
The dendrites may be melanocyte dendrites.

The present invention also relates to a dendrite control method characterized by applying an extract of Rosaceae curin. The dendrite control method is preferably a method used for a dendrite control test or a method used for cells or tissues taken out of the body or artificially produced.
Moreover, this invention provides the screening method of the dendrite positive control agent using the extract of a Rosaceae karin.

Here, dendrite control of the present disclosure refers to positive / negative control of dendrite. The negative control refers to control in a direction that suppresses the activity among the life phenomena, and the positive control refers to control in the direction that promotes the activity among the life phenomena.
Examples of dendrite negative control include suppression of dendrite formation, regression of dendrite, suppression of dendrite activation, and the like. The dendrite positive control includes, for example, that the formation of dendrite is promoted, the dendrite is elongated, and the dendrite is activated.

According to the present invention, it is possible to provide an excellent dendrite negative control agent and a screening method for a dendrite positive control agent using the same.

It is a photograph figure which shows the formation condition of the melanocyte dendrite after adding alpha-melanocyte stimulating hormone 30 minutes after adding a karin extract, and culturing for 3 days. Note that 0 (control), 0.1, 0.3, and 0.5 vol% in the activated state are final concentrations of karin. The length of one bar is 100 μm. It is a figure which shows the formation inhibitory effect of the dendritic process of a melanocyte after adding alpha-melanocyte stimulating hormone 30 minutes after adding a karin extract, and culture | cultivating for 3 days. The vertical axis shows the proportion of dendritic cells in living cells. Note that 0 vol% (no αMSH added), 0 (control) in the activated state, 0.1, 0.3, and 0.5 vol% are the final concentrations of karin. The left figure is a photograph showing the formation of dendrites of melanocytes after 3 days of culture after adding α-melanocyte-stimulating hormone 30 minutes after addition of the cassis extract. The right figure is a figure which shows the formation condition of the dendrites of a melanocyte after adding (alpha) -melanocyte stimulating hormone after adding Sempukuka extract and culturing for 3 days. Note that 0 (no αMSH added), 0 (control of the activated state), and 1.0 vol% are the final concentrations of the cassis extract and Sempukuka extract, respectively. The length of one bar is 100 μm. The left figure is a figure which shows the formation inhibitory effect of the dendrites of a melanocyte after adding the alpha-melanocyte stimulating hormone 30 minutes after adding a cassis extract, and culture | cultivating for 3 days. The right figure is a figure which shows the formation inhibitory effect of the dendrites of a melanocyte after adding the alpha melanocyte stimulating hormone 30 minutes after adding a Sempukuka extract, and culture | cultivating for 3 days. Note that 0 (no αMSH added), 0 (control of the activated state), and 1.0 vol% are the final concentrations of the cassis extract and Sempukuka extract, respectively. It is a photograph figure which shows the retraction state of the dendrite of a melanocyte after adding a karin extract 3 days after adding (alpha) -melanocyte stimulating hormone, and culturing for 3 days. Note that 0 (control), 0.1, 0.3, and 0.5 vol% in the activated state are final concentrations of karin. The length of one bar is 100 μm. The left figure is a figure which shows the regression effect of the dendrites of a melanocyte after adding the alpha melanocyte stimulating hormone 3 days after adding a karin extract and culturing for 3 days. The vertical axis shows the proportion of dendritic cells in living cells. Note that 0 (control), 0.1, 0.3, and 0.5 vol% in the activated state are final concentrations of karin.

The kind and production area of the Rosaceae Karin used in the present disclosure are not particularly limited. There are several species of plant called Karin in Japan. There are also several theories on botanical classification, but all of these can be used. Specifically, the plant called Karin covers all species of the genus Rosaceae, the genus Karin, and the quince, and is a concept that includes all of them. These are commonly characterized by having a single flower at the tip of the new shoot, and the yellow, pear-shaped fruit with aroma is not so easy to eat due to its strong acidity, hard fiber and stone cells. In addition to being processed into fruit liquor, pickled syrup, candy, jam, etc., this fruit is used for cough, diuresis, etc. under the names of herbal medicines such as mokka, wamokka, meisa.
Botanical names of the genus Chonomeles, Japanese name Bokeh English name Flowering Quince, scientific name Chaenomeles speciosa (or C. lagenaria), Japanese name quince bokeh English name Japanese quince Scientific name Chaenomeles japonica, Japanese name Chaenomeles sinensis (Thouin Koehne) Chaenomeles
There are sinensis and so on. Karin (Pseudocydonia) plants are classified by CK Schneider as one genus and one species. The Japanese name Karin Scientific name Pseudocydonia
sinensis is included. The genus Cydonia includes the Japanese name Quince English name Cydonia oblonga Miller and the Japanese name Karin scientific name Cydonia sinensis Thouin.
Of these roses, the scientific name Chaenomeles
sinensis (Thouin) Koehne, Chaenomeles sinensis, Pseudocydonia sinensis, Cydonia
Karin represented by any of sinensis Thouin is preferred.

  Any part may be used as the part used by the Rosaceae Karin. As the said site | part, the 1 type (s) or 2 or more types chosen from a leaf, a stem, a petal, a fruit, a seed, a rhizome, a root, a trunk, etc. can be used, for example. Of these, fruits are preferred.

  The method for preparing the Rosaceae Karin is not particularly limited. For example, any one or more of the Rosaceae Karin flowers, leaves, xylem, fruits, roots, etc. (hereinafter referred to as “original”) is dried or dried. After cutting without cutting, it is obtained by extraction with a solvent at a low temperature (for example, less than 4 ° C.) or a normal temperature (for example, 4 to 40 ° C.) to a heating (for example, 40 to 100 ° C.).

Solvents used for extraction are generally water, lower monohydric alcohols (methanol, ethanol, 1-propanol, 2-propanol, 1-butanol, 2-butanol, etc.), liquid polyhydric alcohols (propylene glycol, 1,3-butylene glycol, etc.), lower esters (ethyl acetate, etc.), hydrocarbons (benzene, hexane, pentane, etc.), ketones (acetone, methyl ethyl ketone, etc.), ethers (diethyl ether, tetrahydrofuran, dipropyl ether) And acetonitrile. In addition, it is preferable that the number of carbons in "lower" is 1-4.
As an example of a preferable extraction method, ethyl alcohol or 1,3-butylene glycol having a water content of 0 to 100 vol% (preferably a water content of 50 to 100 vol%) is used, and 1 to 1 at room temperature (about 4 to 40 ° C.). An example is a method in which after 5 days of extraction, filtration is performed, and the obtained filtrate is further left to mature for about 1 week, followed by filtration again. At the time of filtration, impurities such as impurities and colored substances may be removed using a filtering agent such as activated carbon. The water concentration 100 vol% is water.

  The extract of the Rosaceae curin may be used as an active ingredient as it is, and if necessary, it is used after removing the extraction solvent, purifying treatment such as filtration, deodorization such as ion exchange resin, decolorization, etc. Also good. It is also possible to obtain a highly active fraction or the like by using a separation and purification means such as liquid chromatography.

  The Rosacea quince extract can be used as it is by mixing the extract alone or with the extract obtained by different extraction methods, or by diluting, concentrating or drying the extract, to obtain a liquid, powdery form. Alternatively, it can be prepared in the form of a paste and used.

By the way, since dendrites are thought to be involved in some kind of delivery between cells, the mechanism of dendrite formation has been studied also in the pharmaceutical field and the cosmetic field. For example, Purkinje cells and melanocytes are known as cells having dendrites, but by controlling these dendrites, prevention of symptoms or conditions caused by the formation of these dendrites, It can be used for treatment or improvement.
For example, Purkinje cells are cells involved in the nervous system, but if there is an agent that can control the dendrite of Purkinje cells, the agent can also be used for nerve cell regeneration and the like. In addition, when melanin production increases, melanin migration to keratinocytes can be suppressed by negatively controlling the dendrites of melanocytes, and finally pigmentation such as spots and buckwheat is suppressed. It is also possible to use it as an agent.

And, as shown in the Examples below, in the dendrite formation inhibition test and dendrite retraction test after dendrite formation, the extract of the Rosaceae Karin of the present disclosure has a dendrite formation inhibitory action, a dendritic shape It was recognized that negative control of dendrites such as protrusion retraction action is possible.
Therefore, since the extract of the rose family karin of the present disclosure has a dendrite negative control (for example, dendrite formation inhibitory action, dendrite retraction action, etc.), the rose family karin extract is contained. It can be used as a dendrite negative control agent (for example, dendrite formation inhibitor, dendrite retraction agent, etc.) as an active ingredient.
The extract of the Rosaceae curin of the present disclosure may be used for dendritic negative control (for example, dendrite formation inhibitory action, dendritic retraction action, etc.), and a dendritic negative control agent ( For example, it can be used for various preparations intended for use as described above, such as dendrite formation inhibitors, dendritic retraction agents, etc., and can also be used to produce these various preparations. is there.
The extract of the rose family karin of the present disclosure has dendritic negative control (dendritic formation inhibitory action, dendritic retraction action, etc.), and prevents, improves, and improves various symptoms and conditions caused by dendritic formation. And / or can be used in methods for treatment. Therefore, the rose carin extract of the present disclosure is ingested or administered to animals including humans and used in a method for preventing, ameliorating and / or treating various symptoms and conditions due to dendrite formation. It is possible.

Therefore, the rose curin extract of the present disclosure is suitable for skin denaturation and cosmetics (preferably whitening cosmetics) for dendritic negative control (dendrite formation inhibitory action, dendritic regression action, etc.). ), Pharmaceuticals, quasi-drugs, foods and functional foods (for example, foods for specified health use) and the like, and the preparations of the present disclosure are useful as these skin external preparations, cosmetics and the like.
The rose curin extract of the present disclosure is particularly suitable for use in external preparations for skin, cosmetics (preferably whitening cosmetics), quasi-drugs, pharmaceuticals, and the like. Is preferred. Examples of the external preparation for skin and cosmetic include skin lotion, emulsion (oil-in-water type, etc.), cream (water-in-oil type, etc.), pack cosmetic, liquid foundation, ointment, hair nourishing agent and the like.

  The content of the rose carin extract of the present disclosure is preferably 0.00001 to 5% by mass, more preferably 0.0001 to 2% by mass as a dry solid content in the preparation. If it exists in this range, this plant extract can be mix | blended stably and the outstanding dendrite negative control effect | action can be exhibited. Moreover, when using an extract, if the content of the dry solid content which is a solute is in the said range, the extract concentration will not be limited at all.

The rose carin extract of the present disclosure is used in combination with a drug that exhibits a whitening effect by a mechanism other than dendrite control, such as a tyrosinase inhibitor, a melanin production inhibitor, an anti-inflammatory agent, and a melanin excretion promoter. It is preferable to do this. Thereby, it becomes possible to exhibit a synergistic whitening effect.
Examples of tyrosinase inhibitors and melanin production inhibitors include L-ascorbic acid and its derivatives, and salts thereof (ascorbic acid-2-glucoside, magnesium ascorbyl phosphate, sodium ascorbyl phosphate, 3-O-ethylascorbic acid Etc.), hydroquinone and derivatives thereof (arbutin etc.), tranexamic acid and derivatives thereof (cetyl tranexamic acid, trans-4-aminomethylcyclohexanecarboxylic acid methylamide etc.), salicylic acid and derivatives thereof (4-methoxysalicylic acid etc.) and salts thereof Ellagic acid and its derivatives, kojic acid and its derivatives, adenosine-1-phosphate, linoleic acid, 5,5′-dipropyl-biphenyl 2,2′-diol, 4- (4-hydroxyphenyl) -2-butanol Or a derivative thereof, Zorushin and derivatives thereof (4-n-butyl resorcinol and the like), placental extract, chamomile extract, but nicotinamide, and the like, ascorbic acid 2-glucoside, 3-O-ethyl-ascorbic acid, or arbutin is preferred. Anti-inflammatory agents include tranexamic acid and its derivatives, glycyrrhizic acid derivatives such as dipotassium glycyrrhizinate, glycyrrhetinic acid derivatives such as stearyl glycyrrhetinate, salicylic acid and its derivatives, chamomile extract, etc., but tranexamic acid, dipotassium glycyrrhetinate or Stearyl glycyrrhetinate is preferred. Examples of the melanin excretion promoter include nicotinic acid derivatives such as nicotinic acid amide, adenosine-1-phosphate and derivatives thereof, and nicotinic acid amide or adenosine-1-phosphate is preferable.

In addition, you may use the said formulation in combination with arbitrary components other than the extract of the rose family karin of this indication as needed. Other components may be pharmaceutically acceptable components, such as cell activators, antioxidants, humectants, UV inhibitors, solvents (water, alcohols, etc.), oils, surfactants, Thickener, powder, chelating agent, pH adjuster, emulsifier, stabilizer, colorant, brightener, flavoring agent, flavoring agent, excipient, binder, disintegrant, lubricant, diluent, penetration A pressure regulator, a fragrance | flavor, etc. are mentioned, What is necessary is just to mix | blend according to the formulation for which these are aimed.
The form of the preparation is not particularly limited, and may be any form such as liquid, paste, gel, solid, and powder.

  In addition, the rose curin extract of the present disclosure can be used in a dendrite control method, and the dendrite control method can also be used in a dendrite control test and is collected from an animal. Using raw dendritic cells as raw materials, pharmaceuticals (cell medicines, etc.); medical materials (artificial substitutes or substitutes, etc.); methods for producing these intermediate products or for analyzing them It is also possible to use this method.

  In addition, since the extract of the Rosaceae Karin of the present disclosure negatively controls dendrites, the Karin extract can be used in a screening method for an agent capable of positively controlling dendrites. . Thereby, it is possible to screen a dendrite controlling agent or a substance having a dendrite controlling action or to evaluate dendrite control. It is also possible to search for substances having different action mechanisms by adding the test substance and the karin extract at the same time or separately.

When dendrites are elongated compared to the state in which the extract of the Rosaceae curin and the test substance of the present disclosure are added and the state where dendrites are not formed or the state where dendrites are negatively controlled Alternatively, when dendrites are positively controlled, such as when formation of dendrites is promoted, the test substance can be determined as a dendritic positive control agent. In addition, it is also possible to make dendrites negatively controlled by adding an extract of a rose family karin of the present disclosure and a known dendritic positive control agent.
In addition, the dendritic formation state when adding the extract of the Rosaceae curin of the present disclosure and a known dendrite positive control agent, and the dendrite when adding the dendritic positive control agent and the test substance In contrast, when dendrites are more negatively controlled than when the extract of the rose family karin of the present disclosure is added, the test substance is judged as a good dendritic negative inhibitor be able to.

Although the said determination method is not specifically limited, The example is given to the following.
For example, a photo of cell morphology was taken with a phase contrast microscope (Olympus), and the number of dendritic cells (cells in which one or more dendrites were newly formed from bipolar cells) were determined. Count and calculate the number of dendritic cells / total number of cells × 100%. The positive / negative of the dendritic control of the test substance is judged by comparison with the control.

Below, a suitable example of the search method of the dendrite positive control agent using the extract of the Rosaceae curin of this indication is given.
Screening a dendrite formation promoter by culturing melanocytes in the presence of an extract of the Rosaceae curin of the present disclosure, adding a test substance thereto, and observing dendritic morphology changes of the melanocytes Can do.
At this time, a melanocyte activator may be added. Examples of the melanocyte activator include α-melanocyte stimulating hormone (α-MSH), stem cell growth factor (SCF).
cell factor), endothelin-1 and the like.
In the case of melanocytes, transfer to melanosomes to keratinocytes is promoted, and therefore can be applied to substance search such as improvement of gray hair.

The culture conditions in the above-mentioned dendrite control method, dendrite control agent screening method and dendrite control evaluation method may be the known culture conditions, medium conditions, etc. according to the cells having dendrites to be used. Good.
For example, when normal melanocytes are used, the culture may be performed under known culture conditions in which normal melanocytes can be cultured. As a culture growth medium, a commercially available melanocyte medium or the like may be used.
The culture temperature should just be the temperature which can be cultured corresponding to the origin of the melanocyte to be used, for example, 20 degreeC-40 degreeC, preferably about 37 degreeC in the case of human origin. The carbon dioxide concentration may be about 5 vol%.

  In addition, when Purkinje cells are used, Purkinje cells are cultured by a known culture method in the presence of the extract of the Rosaceae curin of the present disclosure, and evaluation of positive control of dendrites is performed. It can be applied to substance search such as nerve regeneration promotion.

In addition, this technique can also employ | adopt the following structures.
[1] A dendritic negative control agent comprising an extract of rose family karin as an active ingredient.
[2] The dendrite negative control agent according to the above [1], which is an extract from a fruit of a rose family karin.
[3] The dendrite negative control agent according to [1] or [2], wherein the extract is extracted with water, alcohols, or hydrous alcohols.
[4] The dendritic negative control agent according to any one of [1] to [3], wherein the dendritic negative control is melanocyte dendritic negative control.
[5] The dendrite negative control according to any one of [1] to [4], wherein the dendrite negative control is dendrite formation suppression, dendrite retraction, or dendrite activation suppression. Control agent.
[6] A method for controlling dendrites, characterized by applying an extract of a rose family karin.
[7] The dendrite control method according to the above [6], which is an extract of the Rosaceae curin according to any one of the above [1] to [3].
[8] A screening method for a dendrite controlling agent using an extract of a rose family karin.
[9] The screening method for a dendrite controlling agent according to [8] above, which is an extract of the Rosaceae curin according to any one of [1] to [3].

  Examples, Reference Examples, Comparative Examples, Test Examples, Production Examples, etc. are given below to describe the present invention (present technique) more specifically. However, the present invention (present technique) is not limited to these examples. It is not something.

<Manufacture example 1: Manufacture of 50% of karin fruit ethanol extract>
100 g of ethanol / water mixed solution containing 50 vol% ethanol was added to 10 g of dried fruit of Chalinomeles sinensis, extracted at room temperature for 3 days, and then filtered to obtain 50 vol% ethanol extract of karin. The dry solid content of this extract was 3.0%.

<Production Example 2: Production of Karin Fruit Purified Water Extract>
Purified water extract was obtained by adding 100 mL of purified water to 10 g of dried fruit of Chalinomeles sinensis (Thouin) Koehne, followed by extraction at room temperature for 3 days, followed by filtration. The dry solid content of this extract was 4.5%.

<Production Example 3: Production of 99.5 vol% ethanol extract of karin fruit>
To 10 g of immature fruits of Karin (Pseudocydonia sinensis) was added 100 mL of 99.5 vol% ethanol, and extraction was performed at room temperature for 3 days, followed by filtration to obtain 99.5 vol% ethanol extract of Karin fruits. The dry solid content of this extract was 0.7%.

<Production Example 4: Manufacture of quince fruit 50 vol% ethanol-containing extract>
To 10 g of dried quince (Cydonia oblonga) fruit, 100 mL of ethanol / water mixed solution containing 50 vol% ethanol was added and extracted at room temperature for 3 days, followed by filtration to obtain 50 vol% ethanol extract of quince fruit. The dry solid content of this extract was 4.4%.

<Production Example 5: Manufacture of 50 vol% ethanol-containing extract of crisp fruit>
To 10 g of dried fruit of Chaenomeles japonica, 100 mL of an ethanol / water mixed solution containing 50 vol% ethanol was added, followed by extraction at room temperature for 3 days, followed by filtration to obtain a 50 vol% ethanol extract of quince fruit. The dry solid content of this extract was 1.2%.

<Production Example 6: Production of Karin Flower 50vol% ethanol-containing extract>
100 g of ethanol / water mixed solution containing 50 vol% ethanol is added to 10 g of dried flower of Chalinomeles sinensis (Thouin) Koehne, extracted at room temperature for 3 days, filtered to obtain 50 vol% ethanol extract of karin flowers. Obtained. The dry solid content of this extract was 2.4%.

<Test Example 1: Melanocyte dendrite formation inhibition test>
[Example 1: Karin extract]
The melanocyte dendrite formation inhibitory action was investigated using the Karin extract produced in Production Example 1 above.
Specifically, an appropriate amount of epidermal melanocyte (melanocyte) medium containing HMGS growth additive is added to a 6-well plate, 1 × 10 ⁵ normal human epidermal melanocytes (melanocytes) are seeded, and the carbon dioxide concentration is 37 ° C. Static culture was performed in 5 vol%. After 2 days of culturing, the medium was replaced with a medium in which BPE was removed from the HMGS growth additive, and the content of the karin extract produced in Production Example 1 in the medium was 0 (control in activated state), 0.1,. After 30 minutes of adjusting to 3,0.5 vol%, α-MSH was added and mixed so that the final concentration was 10 ⁻⁸ mol / L.
Three days later, a photograph of the cell morphology was taken with a phase-contrast microscope (Olympus), and the number of dendritic cells (cells in which one or more dendrites were newly formed from bipolar cells) And the number of dendritic cells / total number of cells × 100 was calculated, and the results are shown in FIG. 1, Table 1, and FIG.
[Reagents]
Epidermal melanocytes (melanocytes) basal medium: Medium 254 medium; sterile liquid medium M-254-500 (manufactured by Life Technologies) for human epidermal melanocyte culture
HMGS growth additive: HMGS growth additive dispensing kit KM-6350 (manufactured by Kurashiki Boseki Co., Ltd.)
Normal human epidermal melanocytes (Human Epidermal Melanocytes),
neonatal (HEMn-LP); newborn lightly pigmented donorC-002-5C (Life Technologies)
α-MSH: α-melanocyte stimulating hormone M4135 (manufactured by Sigma)

(Comparative example 1) The melanocyte dendrite formation suppression test was done like [Example 1].
However, the “karin extract” of the sample was replaced with “cassis extract” and added so that the content in the medium was 1.0 vol%. The cassis extract was obtained by adding 50 g of purified water to 10 g of cassis (Ribes nigrum L.) fruit, followed by heating at 50 ° C. for 3 days, followed by filtration. The dry solid content of this extract was 3.1%.
(Comparative example 2) The melanocyte dendrite formation suppression test was done like [Example 1].
However, “Karin extract” of the sample was replaced with “Sempukuka extract” and added so that the content in the medium was 1.0 vol%. The Sempukuka extract was obtained by adding 500 g of an ethanol / water mixture containing 50 vol ethanol to 10 g of flowers of Sempukuka: Oguruma (Inula Britannica linne 'var chinensis), followed by extraction at room temperature for 3 days, followed by filtration. The dry solid content of this extract was 0.3%.
The results of Comparative Examples 1 and 2 are shown in FIGS.

<Test Example 2: Melanocyte Dendritic Retraction Test>
[Example 2: Karin extract]
The test was performed in the same manner as in Test Example 1 except that the order and schedule of addition / observation of α-MSH • kaline extract were changed. Three days after adding α-MSH as an activator, it was confirmed that melanocyte dendrites had been sufficiently formed, and then a karin extract was added, followed by further culturing for 3 days. The result is shown in FIG.
Even when the dendrites of the cells were already activated and elongated, it was observed that the dendrites that had been elongated were regressed by adding the karin extract. The results of calculating the number of dendritic cells / total number of cells × 100 are shown in Table 2 and FIG.

Cassis extract and Sempukuka extract are known to have a whitening effect, but it was confirmed that even when added to a high concentration of 1.0 vol%, there is no dendrite formation inhibitory effect. In contrast, the Karin extract was found to have a negative control action on dendrites. Specifically, when a dendritic activator was added at the same time as the karin extract, formation or elongation of dendrites was suppressed.
In addition, it was recognized that the dendrites that had been elongated were retracted even when the karin extract was added after the dendrites activator was first added to activate and extend the dendrites. In particular, retraction of the elongated dendrites makes it possible to develop a whitening effect even if the amount of melanin produced is large, and to improve already formed pigmentation. Such negative control of dendrites is considered to be an action different from a tyrosinase inhibitory action, a melanin production inhibitory action, an anti-inflammatory action, a melanin excretion promoting action, etc., which have been conventionally searched for as a whitening effect.
Therefore, the Karin plant extract of the present disclosure and a drug that exhibits a whitening effect by an action mechanism other than dendrite control (eg, tyrosinase inhibitor, melanin production inhibitor, anti-inflammatory agent, melanin excretion promoter, etc.) In combination with, there is a high possibility of producing a synergistic whitening effect.
In addition, the action of suppressing the exchange of substances between cells via dendrites may lead to new cosmetic uses and pharmaceutical uses. It is considered promising as a method for searching for new whitening agents, white hair improving agents, nerve regeneration promoting agents, etc., that is not directed to suppressing the amount of melanin synthesis as in the past.

[Prescription Example 1: Solubilized lotion]
(Ingredient) (mass%)
1. POE (40 mol) hydrogenated castor oil 0.5
2. POE (12 mol) dioleate 0.3
3. Astaxanthin 0.05
4.1,3-Butylene glycol 2.0
5. Glycerin 2.0
6). Ethanol 15.0
7). Tranexamic acid 2.0
8). Karin extract of Production Example 1 0.1
9. Sodium lactate 0.2
10. Methyl paraoxybenzoate 0.1
11. Purified water balance (production method)
A. Components 1 to 6 are mixed and dissolved.
B. Components 7 to 11 are mixed and dissolved.
C. A was added to B to obtain a skin lotion.

[Formulation Example 2: Emulsion lotion]
(Ingredient) (mass%)
1. Soybean-derived hydrogenated phospholipid 0.5
2. Cetostearyl alcohol 0.1
3. Polyoxyethylene (10 mol) cholesterol ether 0.2
4). Acetic acid-dl-α-tocopherol 0.1
5. Squalane 0.1
6). Hydroxyethyl cellulose 0.03
7). Purified water remaining amount 8. Stearyl glycyrrhetinate 0.25
9. Karin extract of Production Example 2 0.02
10. Disodium monohydrogen phosphate 0.1
11. Monosodium dihydrogen phosphate 0.1
12 Glycerin 3.0
13. Dipropylene glycol 2.0
14 Ethanol 7.0
15. Perfume appropriate amount (production method)
A. Ingredients 1-5 are heated to 75 ° C. and mixed and dissolved uniformly.
B. Ingredients 6 and 7 are heated to 75 ° C. and mixed and dissolved uniformly. Add B to A and emulsify.
D. C was cooled, components 8 to 15 were added, and an emulsified lotion was obtained.

[Prescription Example 3: Latex]
(Ingredient) (mass%)
1. Polyoxyethylene (20) sorbitan monooleate 1.0
2. Sorbitan sesquioleate 0.5
3. Glyceryl trioctanoate 0.5
4). Jojoba oil 0.5
5. Squalane 0.5
6). 6. Purified water remaining amount Edetate disodium 0.1
8). Methylparaben 0.2
9. Phenoxyethanol 0.5
10. Glycerin 5.0
11. Propanediol 1.0
12 Propylene glycol 2.0
13. Sodium lactate 0.5
14. 2-O-α-D-glucosyl-L-ascorbic acid (Note 1) 1.0
15. Karin extract of Production Example 3 0.005
16. Xanthan gum 0.05
17. Purified water 10.0
18. Ethanol 3.0
19. Perfume appropriate amount (Note 1): Hayashibara Co., Ltd. (production method)
A: Swell component 16 with component 17 heated to 70 ° C.
B: Components 1 to 5 are heated and mixed at 70 ° C.
C: Components 6 to 13 are heated and dissolved at 70 ° C., then added to B and emulsified.
D: After cooling C to room temperature, ingredients 14, 15, 18 and A were added to obtain a cosmetic liquid.
The emulsion of this Formulation Example 3 moisturized the skin, prevented the skin from drying for a long time, and was excellent in the moisturizing effect.

[Formulation Example 4: Hair Nourishing]
(Ingredient) (mass%)
1. Srutianin 1.5
2. Ginkgo biloba extract 0.5
3. Tranexamic acid 1.0
4). Quince extract of Production Example 4 1.0
5. Glycerin 2.0
6). 6. Purified water remaining amount D-pantothenyl alcohol 0.3
8). Hinokitiol 0.02
9. Cephalanthin 0.001
10. Tocopherol acetate 0.01
11. L-Menthol 0.2
12 Polyoxyethylene hydrogenated castor oil 0.2
13. Ethanol 60
(Production method)
A. Components 1 to 6 are mixed and dissolved.
B. Components 7 to 11 are mixed and dissolved.
C. B was added to A to obtain a hair nourishing agent.

[Prescription Example 5: Ointment A]
(Compounding ingredients) (mass%)
1. Stearyl alcohol 18.0
2. Owl 20.0
3. Polyoxyethylene (20) monooleate 0.25
4). Glycerin monostearate 0.3
5. Vaseline 40.0
6). 6. Purified water remaining amount Glycerin 10.0
8). Adenosine-1-phosphate 0.5
9. Ribbed extract of Production Example 5 1.0
(Production method)
A. 1-5 are mixed uniformly at 70 degreeC.
B. Warm 6-8 to 70 ° C.
C. Add B to A and emulsify.
D. C was cooled and 9 was added to obtain an ointment.

Claims (5)

Scientific name Chaenomeles sinensis (Thouin) Koehne, Chaenomeles sinensis, Pseudocydonia sinensis, dendritic retraction agent to Karin extract the active ingredient is either Cydonia sinensis Thouin (However, inhibiting the tyrosinase activity inhibition, or melanin Excluding whitening agents due to suppression.) Wherein the extract, dendritic retraction agent according to claim 1, wherein an extract from the fruit. Before Ki抽 distillate is water, alcohols, dendritic retraction agent according to any one of claims 1 to 3, which has been extracted in those selected from water-containing alcohols. The dendrites regression, dendritic retraction agent of any of claims 1-4 melanocyte dendrites retraction. A screening method for a dendrite control agent, comprising a step of reducing a dendrite by an extract of karin that is one of the scientific names Chaenomeles sinensis (Thouin) Koehne, Chaenomeles sinensis, Pseudocydonia sinensis, Cydonia sinensis Thouin .