JP2017048155A - Dkk1 expression promoting agent and whitening agent - Google Patents

Abstract

PROBLEM TO BE SOLVED: To provide DKK1 expression promoting agents containing as an active ingredient what has DKK1 expression promoting action found out of ingredients derived from natural products excellent in safety, and to provide whitening agents based on the DKK1 expression promoting action of the ingredients derived from natural products.SOLUTION: As an active ingredient of the DKK1 expression promoting agent and the whitening agent based on DKK1 expression action, an extract from one or more natural products selected from the group consisting of Alpinia speciosa leaf, Glycyrrhiza uralensis leaf, Ziziphus jujuba, royal jelly, and Eriobotrya japonica is contained.SELECTED DRAWING: None

Description

  The present invention relates to a DKK1 expression promoter and a whitening agent.

  The Wnt (Wind) signal pathway is an intracellular signal pathway that is widely conserved from nematodes to mammals. In the Wnt signal pathway, signals are transmitted by binding Wnt, which is a secreted protein, to Wnt receptors present on the cell membrane. In the epidermis, it is known that activation of the Wnt signal pathway promotes melanocyte differentiation / proliferation and melanin synthesis.

  On the other hand, DKK1 (Dickkopf 1), which is a secreted protein that inhibits the binding between Wnt and the Wnt receptor, is known as a factor that inhibits the Wnt signal pathway. It is known that DKK1 is secreted from dermal skin fibroblasts in the skin, suppresses the differentiation and proliferation of melanocytes existing in the epidermis, and suppresses melanin synthesis (see Non-Patent Document 1). DKK1 is also known to promote proliferation of keratinocytes (epidermal keratinocytes) in the epidermis and suppress melanin uptake from melanocytes (see Non-Patent Document 2).

  DKK1 is known to be highly expressed in fibroblasts at palmar palms (palms and palms). In general, skin color varies greatly depending on race, but palmar palms may be of any race. The reason for the whiteness is thought to be due to the high expression level of DKK1 in the palmar region.

  Therefore, if the expression of DKK1 can be increased in dermal fibroblasts, it is considered that pigmentation in the epidermis can be suppressed by inhibiting the activation of the Wnt signal pathway.

J. Cell Biol., 2004, vol.165, no.2, pp.275-285 FASEB J., 2008, vol.22, issue 4, pp.1009-1020

  The present invention is based on the DKK1 expression promoting agent having a DKK1 expression promoting action as an active ingredient, and a DKK1 expression promoting action of the natural product-derived ingredient, which are found out of natural product-derived ingredients having excellent safety. The object is to provide a whitening agent.

  In order to solve the above-mentioned problems, the DKK1 expression promoter of the present invention is extracted from one or more natural products selected from the group consisting of moon peach leaf part, licorice leaf part, Taiso, royal jelly, and loquat. It is characterized by containing a product as an active ingredient. Further, the whitening agent of the present invention is based on the DKK1 expression promoting action, and is selected from one or more natural products selected from the group consisting of moon peach leaf part, licorice leaf part, typhoid, royal jelly, and loquat. The extract is contained as an active ingredient.

  According to the present invention, by containing as an active ingredient an extract from one or more natural products selected from the group consisting of moon peach leaves, licorice leaves, peanuts, royal jelly, and loquat It is possible to provide a DKK1 expression promoter and a whitening agent that are excellent in effect and highly safe.

Embodiments of the present invention will be described below.
The DKK1 expression promoter according to the present embodiment contains, as an active ingredient, an extract from one or more natural products selected from the group consisting of moon peach leaves, licorice leaves, lysins, royal jelly, and loquat. Is. In addition, the whitening agent of the present embodiment is based on the DKK1 expression promoting action, and is one or more natural products selected from the group consisting of moon peach leaves, licorice leaves, taisou, royal jelly, and loquat The extract from this is contained as an active ingredient.

  Here, the “extract” in the present embodiment includes an extract obtained using the above natural product as an extraction raw material, a diluted or concentrated solution of the extract, a dried product obtained by drying the extract, or these Both of these crudely purified products and purified products are included.

  Extraction raw materials used in the DKK1 expression promoting agent and whitening agent of this embodiment are Tsukitomo (scientific name: Alpinia speciosa (Wendl.) K. Schum.), Licorice, Tisoiso (herbal medicine name), royal jelly, and loquat (scientific name: Eriobotrya japonica Lindley).

  Moon peach (Alpinia speciosa (Wendl.) K. Schum.) Is a perennial evergreen herb belonging to the genus Glyceraceae and is distributed from southern Kyushu to India and is easily available from these regions . Tsuki-Peach is called Sannin in Okinawa and is used in Muchy, a traditional confectionery since the Ryukyu dynasty. The part used as an extraction raw material is a leaf part.

  Licorice is a perennial herb belonging to the genus Glycyrrhiza. Licorice includes Glychyrrhiza glabra, Glychyrrhiza inflata, Glychyrrhiza uralensis, Glychyrrhiza aspera, Glychyrrhiza aspera, Glychyrrhiza aspera, Glychyrrhiza aspera, Glychyrrhiza aspera, Glychyrrhiza aspera, Glychyrrhiza aspera ), Glychyrrhiza yunnanensis, Glychyrrhiza lepidota, Glychyrrhiza echinata, Glychyrrhiza echinata, Glychyrrhiza acanthocarpa, etc. Licorice may be used as an extraction raw material, especially Glychyrrhiza glabra, Glychyrrhiza uralen sis) and Glychyrrhiza inflata are preferably used as raw materials for extraction.

  In this embodiment, the component part of the licorice used as an extraction raw material is a leaf part. Here, licorice has been used as a raw material for medicinal or sweeteners since ancient times and is known to contain glycyrrhizin and other useful components. Generally speaking, “licorice” refers to its root. However, in this embodiment, as a result of paying attention to the remaining part after using the root part, it has been found that an extract from the licorice leaf part has a DKK1 expression promoting action. Note that glycyrrhizin and other useful components contained in the root are not contained in the licorice leaf.

  Taisou (body name, crude drug name) is a mature fruit part of the Rhamnaceae family jujuba (scientific name: Zizyphus jujuba Miller var.inermis Rehder). Jujube is a deciduous small tree that grows naturally in eastern Asia such as Southern Europe and China, and isopia can be easily obtained from these regions. Taisou is known to have relieving, tonic and diuretic effects and is used for loss of appetite, diarrhea, palpitation and the like.

  Royal jelly is the throat of a young worker bee among the European honeybees (scientific name: Apis melifera L., aka: Apis mellifera) or honeybees (scientific name: Apis indeca Radoszkowski) distributed in Europe, Africa, etc. It is a secretion from the gland that feeds the queen bee larvae and adult queen bees.

  Biwa (Eriobotrya japonica Lindley) is an evergreen tree belonging to the genus Biwa of the family Rosaceae, is native to the southwestern part of China and is cultivated in Nagasaki, Chiba, Kagoshima, etc., and can be easily obtained from these areas. Examples of the part that can be used as the extraction raw material include a leaf part, a trunk part, a branch part, a flower part, a fruit part, a seed part, a root part, or a mixture of these parts, and a leaf part is preferable.

  An extract from the above natural product can be obtained by drying the raw material for extraction and then pulverizing the raw material as it is or using a crusher and subjecting it to extraction with an extraction solvent. Drying may be performed in the sun or using a commonly used dryer. Moreover, after performing pretreatment, such as degreasing, with a nonpolar solvent such as hexane, it may be used as an extraction raw material. By performing pretreatment such as degreasing, extraction with a polar solvent can be performed efficiently.

  As the extraction solvent, it is preferable to use a polar solvent, and examples thereof include water and hydrophilic organic solvents. These are used alone or in combination of two or more at room temperature or a temperature below the boiling point of the solvent. It is preferable.

  Examples of water that can be used as the extraction solvent include pure water, tap water, well water, mineral spring water, mineral water, hot spring water, spring water, fresh water, and the like, and those subjected to various treatments. Examples of the treatment applied to water include purification, heating, sterilization, filtration, ion exchange, osmotic pressure adjustment, buffering, and the like. Therefore, the water that can be used as the extraction solvent in this embodiment includes purified water, hot water, ion exchange water, physiological saline, phosphate buffer, phosphate buffered saline, and the like.

  Examples of hydrophilic organic solvents that can be used as extraction solvents include lower aliphatic alcohols having 1 to 5 carbon atoms such as methanol, ethanol, propyl alcohol, and isopropyl alcohol; lower aliphatic ketones such as acetone and methyl ethyl ketone; 1,3-butylene. Examples thereof include polyhydric alcohols having 2 to 5 carbon atoms such as glycol, propylene glycol and glycerin.

  When using the liquid mixture of 2 or more types of polar solvents as an extraction solvent, the mixing ratio can be adjusted suitably. For example, when a mixed liquid of water and lower aliphatic alcohol is used as an extraction solvent, the mixing ratio of water and lower aliphatic alcohol is preferably 9: 1 to 1: 9 (volume ratio), More preferably, it is 8: 2 to 2: 8 (capacity ratio). In addition, when a mixed liquid of water and a lower aliphatic ketone is used, the mixing ratio of water and the lower aliphatic ketone is preferably 9: 1 to 2: 8 (volume ratio). When using the liquid mixture with a monohydric alcohol, it is preferable that the mixing ratio of water and a polyhydric alcohol is 5: 5 to 1: 9 (volume ratio).

  The extraction treatment is not particularly limited as long as the soluble component contained in the extraction raw material can be eluted in the extraction solvent, and can be performed according to a conventional method. For example, the extraction raw material is immersed in an extraction solvent 5 to 15 times (mass ratio) of the extraction raw material, the soluble components are extracted at room temperature or under reflux, and then filtered to remove the extraction residue. A liquid can be obtained. When the solvent is distilled off from the obtained extract, a paste-like concentrate is obtained, and when this concentrate is further dried, a dried product is obtained.

  Since the extract from the moon peach leaf part, licorice leaf part, tassium, royal jelly or loquat obtained as described above has an excellent DKK1 expression promoting action and whitening action, the DKK1 expression promoting agent and whitening It can be used as an active ingredient of an agent. In addition, in this embodiment, the whitening action which the extract from a moon peach leaf part, a licorice leaf part, a peanut, a royal jelly, and a loquat has is exhibited based on a DKK1 expression promotion effect.

  The DKK1 expression promoter and whitening agent of this embodiment can be used for a wide range of applications such as pharmaceuticals, quasi drugs, and cosmetics.

  In the present embodiment, any one of extracts from moon peach leaves, licorice leaves, lysins, royal jelly, or loquat may be used as the active ingredient, or two or more of these may be used. You may mix and use as said active ingredient. When an extract from moon peach leaf part, licorice leaf part, tassium, royal jelly or loquat is used as the above-mentioned active ingredient, the blending ratio is from moon peach leaf part, licorice leaf part, typhoid, royal jelly and loquat What is necessary is just to adjust suitably according to the grade etc. of the DKK1 expression promotion effect which an extract has.

  The DKK1 expression promoting agent and whitening agent of the present embodiment may be composed only of an extract from moon peach leaf, licorice leaf, typhoid, royal jelly, or loquat, or a mixture thereof, or moon peach leaf, licorice leaf. , An extract from Taiso, royal jelly, loquat, or a mixture thereof.

  The DKK1 expression promoter and whitening agent of the present embodiment is a powder, granule, tablet, liquid, etc. according to a conventional method using a pharmaceutically acceptable carrier such as dextrin, cyclodextrin and any other auxiliary agent. Can be formulated into any dosage form. In this case, as an auxiliary agent, for example, an excipient, a binder, a disintegrant, a lubricant, a stabilizer, a flavoring / flavoring agent, and the like can be used. The whitening agent can be used by blending with other compositions (for example, skin cosmetics, etc.), and can also be used as an ointment, a solution for external use, a patch, and the like.

  When the DKK1 expression promoter and whitening agent of the present embodiment are formulated, the content of the extract from the moon peach leaf part, licorice leaf part, Taisou, royal jelly, loquat or a mixture thereof is not particularly limited. And can be appropriately set according to the purpose.

  In addition, the DKK1 expression promoter and whitening agent of the present embodiment may contain other natural extracts having a DKK1 expression promoting action, if necessary, from a moon peach leaf part, a licorice leaf part, a tassel, a royal jelly, or a loquat. It can mix | blend with an extract or these mixtures, and can be used as an active ingredient.

  Examples of the method for administering the DKK1 expression promoter and whitening agent of this embodiment to a patient include transdermal administration, oral administration, and the like. Depending on the type of disease, a suitable method for its prevention / treatment is selected as appropriate. do it. In addition, the dosage of the whitening agent of this embodiment may be appropriately increased or decreased depending on the type of disease, severity, individual differences among patients, administration method, administration period, and the like.

  The DKK1 expression promoting agent of the present embodiment promotes the expression of DKK1 in skin fibroblasts through the DKK1 expression promoting action possessed by the extract from the active ingredient, moon peach leaf part, licorice leaf part, tassium, royal jelly, or loquat. In addition, the Wnt signal pathway inhibitory action of DKK1 can suppress melanocyte differentiation and proliferation in the epidermis, suppress melanin production, and suppress melanin uptake from melanocytes in keratinocytes (epidermal keratinocytes). be able to. Therefore, according to the DKK1 expression promoting action of the present embodiment, pigmentation in the epidermis can be prevented, treated or improved by promoting DKK1 expression in dermal fibroblasts. Moreover, the above-mentioned abnormal activation of the Wnt signal pathway is considered to be a cause of carcinogenesis in colorectal cancer, and it is known that DKK1 is inactivated in colorectal cancer cells. Therefore, the DKK1 expression promoter of this embodiment can be used for the treatment or improvement of colorectal cancer through inactivation of the Wnt signal pathway by promoting the expression of DKK1. However, the DKK1 expression promoting agent of the present embodiment can be used for all purposes that are meaningful for exerting the DKK1 expression promoting action in addition to these uses.

  The whitening agent of the present embodiment promotes the expression of DKK1 in dermal fibroblasts through the DKK1 expression promoting action of an extract from the active ingredient, moon peach leaves, licorice leaves, lysins, royal jelly, or loquat. Thus, pigmentation in the epidermis can be prevented, treated or ameliorated.

  Here, in addition to those based on melanin production inhibitory action and melanocyte differentiation / proliferation inhibitory action in melanocytes, conventional whitening agents have been proposed such as those based on epidermal keratinocyte proliferation promoting action and melanin uptake inhibitory action . All of these act on the epidermis and suppress pigmentation such as melanin caused by exposure to ultraviolet rays. In contrast, the DKK1 expression promoting agent and whitening agent of the present embodiment act on dermal fibroblasts, and increase melanin irrespective of the presence or absence of ultraviolet light exposure, such as palmar fin sites. It is different from conventional whitening agents in that it can be prevented.

  Since the DKK1 expression promoter and whitening agent of the present embodiment have an excellent DKK1 expression promoting action, they are suitable for blending in, for example, a skin external preparation or food and drink. In this case, an extract from a moon peach leaf part, a licorice leaf part, a Taiso, royal jelly, or a loquat or a mixture thereof may be blended as it is, or a moon peach leaf part, a licorice leaf part, a Taiso, a royal jelly, or a loquat. A DKK1 expression promoter or whitening agent formulated from the above extract or a mixture thereof may be blended.

  Here, the topical skin preparation is not limited in its category, and includes a wide range of skin cosmetics, quasi-drugs, pharmaceuticals, and the like used transdermally. Creams, milky lotions, beauty essences, lotions, packs, foundations, lip balms, bath salts, hair nicks, hair lotions, soaps, body shampoos and the like.

  In addition, food and drink is not limited to food, pharmaceuticals, quasi-drugs, etc. in administrative divisions, but general foods, health foods, functional foods, health functional foods (orally ingested) Foods for specified health use, functional labeling foods, functional nutritional foods), quasi-drugs, and pharmaceuticals.

  Moreover, since the DKK1 expression promoter and whitening agent of the present embodiment have an excellent DKK1 expression promoting action, they can also be suitably used as reagents for research on these action mechanisms.

  The DKK1 expression promoter and whitening agent of the present embodiment are preferably applied to humans, but animals other than humans (for example, mice, rats, It can also be applied to hamsters, dogs, cats, cows, pigs, monkeys, etc.).

  Hereinafter, although a test example is shown and this invention is demonstrated concretely, this invention is not restrict | limited to each following example at all. In the following test examples, freeze-dried products (samples 1 to 5) shown in Table 1 were used as test samples.

[Test Example 1] DKK1 mRNA expression promoting action test The natural product extract (samples 1 to 5) was tested for DKK1 mRNA expression promoting action as follows.

Human normal skin fibroblasts (NB1RGB) were cultured using Dulbecco's MEM (DMEM) medium containing 10% FBS, and then cells were collected by trypsin treatment. The collected cells are diluted with 10% FBS-containing DMEM medium to a cell density of 1.0 × 10 ⁵ cells / mL, and then seeded in 5 mL each in a 60 mm petri dish (manufactured by FALCON) (5 × 10 ⁵ cells / petri dish). ) And cultured overnight.

After culturing, the medium was removed, and 5 mL each of 10% FBS-containing DMEM medium in which the test samples (samples 1 to 5 and sample concentration are shown in Table 2) were dissolved was added to each dish, and the mixture was incubated at 37 ° C. and 5% CO _2. For 24 hours. In addition, it culture | cultivated similarly using 10% FBS containing DMEM culture medium without a sample as control. After culturing, the medium is removed, and total RNA is extracted with ISOGEN (Nippon Gene, Cat. No. 311-02501), and the amount of each RNA is measured with a spectrophotometer so that it becomes 200 ng / μL. Total RNA was prepared.

  Using this total RNA as a template, the expression level of mRNA was measured for DKK1 and GAPDH as an internal standard. Detection was performed by real-time 2 Step RT-PCR reaction using a real-time PCR device Smart Cycler (Cepheid) using a TaKaRa SYBR PrimeScript RT-PCR kit (Perfect Real Time) (Takara Bio, code No. RR063A). The expression level of DKK1 mRNA was determined as a GAPDH value based on the total RNA preparation prepared from cells cultured in “addition of test sample” and “no sample addition”. From the obtained value, the DKK1 mRNA expression promotion rate (%) was calculated by the following formula.

DKK1 mRNA expression promotion rate (%) = A / B × 100
Each term in the formula represents the following.
A: Correction value with addition of test sample B: Correction value without addition of sample Table 2 shows the results.

  As shown in Table 2, moon peach leaf extract (sample 1), licorice leaf extract (sample 2), diatom extract (sample 3), royal jelly extract (sample 4), and loquat extract (sample 5) ) Were confirmed to have excellent DKK1 mRNA expression promoting action.

  Unlike the conventional whitening agent, the DKK1 expression promoter and whitening agent of the present invention act on the dermis, and can greatly contribute to the prevention or improvement of pigmentation such as skin darkening, spots and freckles.

Claims (2)

  A DKK1 expression promoter characterized by containing, as an active ingredient, an extract from one or more natural products selected from the group consisting of moon peach leaves, licorice leaves, typhoid, royal jelly, and loquat.   A DKK1 expression promoting action characterized by containing, as an active ingredient, an extract from one or more natural products selected from the group consisting of moon peach leaves, licorice leaves, typhoid, royal jelly, and loquat Whitening agent based.