CN105997842A - Skin external preparation composition containing ginsenoside Rg1 - Google Patents

Abstract

The invention provides a skin external preparation composition containing ginsenoside Rg1, and the ginsenoside Rg1 can inhibit psoriatin. The composition for external application to the skin of the present invention can be used for the purpose of preventing and alleviating skin diseases or moisturizing the skin. Wherein the skin disease includes atopic dermatitis or psoriasis. The skin external preparation composition of the present invention can prevent the skin keratinization abnormality induced by psoriatin, prevent and alleviate skin diseases, and increase the moisture content of the skin.

Description

Skin external preparation composition containing ginsenoside Rg1

technical field

The present invention relates to a skin external preparation composition containing ginsenoside Rg1.

Background technique

Everyone wishes to have baby-like radiant and beautiful skin, but the skin gradually loses its elasticity and becomes damaged as it is exposed to various external environments and as we age. The skin is the first barrier for the human body to directly contact with the external environment, and it needs to always directly bear the stimulation brought by the external environment, so very small stimulation may also cause sensitive reactions and changes of the skin. This skin change can lead to different substances than usual, and when exposed to irritating environments periodically and continuously, it may even cause irreversible damage.

Recently, many studies have been actively conducted on psoriasis, an antimicrobial peptide substance in the skin. Especially after it was discovered that in addition to the currently known antibacterial effects of psoriasis, it may also induce abnormal differentiation of skin keratinocytes and thus cause damage to the skin barrier, psoriasin (psoriasin) is considered to be directly related to skin diseases. related important substances. According to the results of an actual study (Regine Glaser, The Journal of investigative dermatology (2009), v.129no.3, p.641-649), it can be known that in patients with typical skin diseases, i.e. atopic dermatitis or psoriasis In the epidermis, very high concentrations of psoriasitin can be detected. This kind of skin with excessive psoriasis concentration will show a structure that is not dense, sparse and damaged, and at the same time, the thickness of the epidermis will be abnormally thickened.

In addition, the present inventors have found through continuous research on psoriasis that when external stimuli such as ultraviolet rays, staphylococci, and retinoic acid are applied, the concentration of psoriasis will also increase. In addition, it has been confirmed that psoriasis is expressed outside the epidermal cells, thereby inhibiting the expression of keratin 1 or 10 among normal epidermal structural proteins, and at the same time inhibiting the expression of keratin 6 or keratin 6 or keratin 10, proteins that cause abnormal epidermal differentiation and proliferation. The production of keratin 16 results in a boost. Therefore, by effectively inhibiting psoriasis, an effect of improving skin characteristics can be obtained.

prior art literature

patent documents

(Patent Document 1) External Skin Composition Containing Ginsenoside Rg3 (Republic of Korea Laid-Open Patent No. 10-2014-0126892)

Contents of the invention

The object of the present invention is to provide a skin external preparation composition containing ginsenoside Rg1, thereby inhibiting the action of psoriasis existing in the skin.

Another object of the present invention is to prevent and alleviate skin diseases such as atopic dermatitis and psoriasis, and to improve the moisturizing power and recovery power of the skin through the above-mentioned inhibitory effect on psoriasis.

In order to achieve the above object, the present invention provides a skin external preparation composition, the active ingredient of which contains ginsenoside Rg1, and the above ginsenoside Rg1 can inhibit psoriasis.

According to the embodiment of the present invention, the structure of the above-mentioned ginsenoside Rg1 is shown in the following chemical formula 1:

<chemical formula 1>

According to an embodiment of the present invention, the content of the above-mentioned ginsenoside Rg1 is 0.001-5% by weight of the whole composition.

According to an embodiment of the present invention, the composition includes more than one of a dermatological composition, a cleansing composition and a cosmetic composition.

The composition of the present invention can be used for the prevention and alleviation of skin diseases or for skin moisturizing purposes. At this time, the aforementioned skin diseases include atopic dermatitis or psoriasis.

The skin external preparation composition containing ginsenoside Rg1 of the present invention can inhibit the action of psoriasis present in the skin.

Therefore, the skin external preparation composition of the present invention can inhibit the abnormal phenomenon of skin keratinocytes induced by psoriasis, restore the damaged skin barrier, prevent and alleviate skin diseases, and at the same time improve the moisturizing power and recovery power of the skin.

Description of drawings

Fig. 1 is a graph showing the mRNA expression level of psoriasis under external stimuli such as ultraviolet B, staphylococcus, and retinoic acid.

Fig. 2a is a graph showing the mRNA expression levels of keratin 1 and keratin 10 under the external stimulation of ultraviolet A and ultraviolet B.

Fig. 2b is a graph showing the mRNA expression levels of keratin 1 and keratin 10 under the external stimulation of staphylococcus.

Fig. 2c is a graph showing the mRNA expression levels of keratin 1 and keratin 10 under the external stimulation of retinoic acid.

Fig. 3a is a graph showing the mRNA expression level of keratin 1 under the action of psorigin and the result of western blot showing the protein expression level.

Fig. 3b is a graph showing the mRNA expression level of keratin 10 under the action of psoriasis and the result of Western blot showing the protein expression level.

Fig. 4 is a graph showing the mRNA levels of psoriasis expressed under the action of interleukin-6 (IL-6), ginsenoside Rg1, and IL-6+ginsenoside Rg1, respectively.

Fig. 5a is a graph showing the amount of mRNA of keratin 1 expressed under the action of psoriasis and psoriasis + ginsenoside Rg1 respectively.

Fig. 5b is a graph showing the amount of mRNA of keratin 10 expressed under the action of psoriasis and psoriasis + ginsenoside Rg1 respectively.

Fig. 6 is an observation photo of keratin 1 expressed by the artificial skin under the action of psoriasis, psoriasis + ginsenoside Rg1 respectively.

Fig. 7 is a graph comparing skin moisture content in the case of using an external preparation for skin containing or not containing ginsenoside Rg1 and using a general external preparation for skin.

detailed description

The inventors of the present invention consider that when a skin external preparation composition is manufactured using a substance that can inhibit the action of psoriasis, it will not only prevent and alleviate various skin diseases, but also further improve the skin's own recovery and moisturizing power, thereby maintaining a more beautiful skin. skin. Therefore, after screening various candidate substances and after repeated failures and unremitting efforts, it was finally found that ginsenoside Rg1 can exert a very good inhibitory effect on the action of psoriasis.

The above-mentioned ginsenoside Rg1 is one of the saponin components contained in the ginseng extract with the structure shown in the following chemical formula 1:

<chemical formula 1>

Ginsenosides are not only different from the saponins contained in more than 750 other plants in terms of chemical structure and pharmacological efficacy, but also have very mild medicinal properties and will not cause any toxicity when used in excess, and will hardly cause any hemolysis. Therefore, It is expected that the skin external preparation composition containing ginsenoside Rg1 of the present invention can be used very safely and effectively.

The above-mentioned ginsenoside Rg1 can be extracted from plants, can also be synthesized and used by corresponding industry-known methods, or can be purchased and used as a commercially available product. Generally, it can be extracted and collected from ginseng. There is no special restriction on the type of ginseng used at this time, and fresh ginseng, red ginseng, white ginseng, Taiji ginseng, tail ginseng, etc. are suitable. In addition, in the above-mentioned extraction process, all parts of ginseng, such as stems, roots, leaves, flowers, and fruits of ginseng, can be used, and the extract is not limited to a specific part.

In the present invention, the content ratio of ginsenoside Rg1 is 0.001-5% by weight of the whole weight of the skin external preparation composition.

The above-mentioned skin external preparation composition comprising ginsenoside Rg1 effectively prevents the abnormal phenomenon of skin keratinocyte formation caused by the above-mentioned psoriasis by inhibiting one of the antibacterial peptides present in the skin, that is, psoriasis, and This prevents or alleviates skin diseases and increases the moisture content of the skin.

In this case, the above skin external preparation composition may further contain a flavonoid.

The composition of the present invention may preferably be at least one of a dermatological composition, a cleansing composition, and a cosmetic composition. In addition, the composition of the present invention may contain ginsenoside Rg1 or an extract containing ginsenoside Rg1 and other additives, and preferably also contain a carrier that is allowed to be used in dermatology and/or cosmetics.

In another embodiment of the present invention, the ginsenoside Rg1 or the extract containing ginsenoside Rg1 can also be converted into a preparation such as a food supplement composition.

There are no particular limitations on the dosage form to which the composition of the present invention is applied, and a dosage form containing a medium or a substrate acceptable for use in cosmetics or dermatology can be used. This includes all dosage forms suitable for topical use, such as solutions, gels, solids, pastes, emulsions obtained by dispersing oils in water, suspensions, microemulsions, microcapsules, microparticles Or in the form of ionic (liposome) and non-ionic small capsule dispersions, or in the form of cream, body lotion, lotion, powder, ointment, spray or concealer stick. Furthermore, it may also take the form of a foam or an aerosol composition comprising a compression accelerator.

These compositions can be produced according to conventional methods in the corresponding fields. More specifically, it can be in the form of ointments, creams, gels, salves, serums, emulsions, aqueous solutions, powders, sticks, and the like.

In addition, the compositions of the present invention may comprise fatty substances, organic vehicles, dissolving agents, concentrating agents, gelling agents, emollients, antioxidants, suspending agents, stabilizers, foaming agents, fragrances, surface active agents agent, water, ionic or nonionic emulsifier, filler, metal ion blocking agent (metal ion blocking agent) such as ethylenediaminetetraacetic acid (EDTA; ethylenediaminetetraacetic acid), chelating agent (chelating agent), preservative, vitamin , blocking agent (blocker), humectant, required oil, dye, pigment, hydrophilic or lipophilic active agent, lipid capsule or any other ingredient frequently used in cosmetics etc. in the field of cosmetics or dermatology Adjuvants used. The added amount of the above-mentioned auxiliary agent is the amount conventionally used in the field of cosmetics or dermatology.

In addition, the composition of the present invention may further contain a skin absorption-promoting substance in order to enhance the skin-improving effect.

In order to facilitate understanding of the present invention, the most representative examples and the like will be described below. The scope of rights of the present invention is not limited thereto and includes all equivalent scopes.

In all drawings used in this specification, ginsenoside RG1 (Ginsenoside Rg1) is represented as G_Rg1, psoriasin (S100A7) is represented as S100A7, keratin 1 (keratin1) is represented as KRT 1, keratin 10 (keratin 10) KRT 10, Staphylococcus aureus as S.aureus, Retinoic acid as RA, Ultraviolet A as UVA, Ultraviolet B as UVB, RT-PCR Corrected reference mRNA (control) for (RT-PCR) or corrected reference protein (control) for Western blotting is denoted as con.

Confirm whether psoriasis induces abnormal keratinization of the skin

In consideration of the fact that psoriasis in the skin maintains a high concentration in skin diseases such as atopic dermatitis and psoriasis, it was confirmed in Experimental Examples 1 to 3 whether external stimuli would also lead to an increase in the concentration of psoriasis in the general skin, and observed The various changes related to the formation of skin keratin that occur when the above-mentioned psoriasis is formed.

The above-mentioned types of external stimuli are ultraviolet A or B, staphylococcus, and retinoic acid.

[Experimental example 1] Confirmation of changes in the expression level of psoriasis under external stimuli

In order to confirm the change in the expression level of psoriasis under external stimulation, first, skin keratinocytes were cultured. As a first step, keratinocytes isolated from the epidermis of newborns were inoculated into KGM medium in a cell culture dish having a diameter of 60 mm at a density of 1.25×10 ⁶ cells/dish. Next, the cell culture dish was stored at 37° C. and 5% CO ₂ in a CO ₂ incubator, and the keratinocytes were cultured to a confluency of about 80%.

Next, the above-mentioned cultured keratinocytes were divided into three experimental groups, and treated with ultraviolet rays B 25mJ/cm ² , Staphylococcus 6μg/ml, and retinoic acid 1μM as external stimuli, and then treated for 2 days. cultivation. Next, the cell-free culture solution was separated, and then the mRNA expression level of psoriasis was measured and quantified by using an enzyme-linked immunosorbent assay (ELISA assay). As correction reference mRNA, the housekeeping gene (housekeeping gene) RPL13A was used.

The above measurement results are shown in FIG. 1 . In all the above experimental groups treated with external stimuli, the mRNA expression of psoriasis all increased.

[Experimental example 2] Confirmation of abnormal skin keratinization induced by external stimulation

In order to confirm whether external stimuli induce abnormal keratinization of the skin, changes in mRNA and protein expression levels of keratin 1 and keratin 10, which are representative epidermal differentiation proteins, were analyzed. The aforementioned keratin 1 and keratin 10 were used as indicators of keratin formation.

The stage of culturing skin keratinocytes was the same as that in Example 1 above. Next, the above-mentioned cultured keratinocytes were divided into several experimental groups, and treated with ultraviolet rays, staphylococci, and retinoic acid as external stimuli. Specifically, after treatment with ultraviolet A 10 mJ/cm ² , ultraviolet B 20 mJ/cm ² , Staphylococcus 3 μg/ml, Staphylococcus 6 μg/ml, and retinoic acid 1 μM, culture was performed for 2 days.

Next, mRNA analysis or protein analysis is performed. The details are as follows.

1) mRNA analysis

After removing the culture medium of the above-mentioned cells, the mRNA of the glial cells was isolated by adding 1 ml of Trizol reagent (Trizol, invitrogen), and then the above-mentioned glial cells were detected by ultraviolet detector (HEWLLETT PACKARD) under the condition of 260nm Cellular mRNA was quantified. Next, the mRNA expression levels of keratin 1 and keratin 10 among glial cell mRNAs were confirmed by reverse transcription-polymerase chain reaction (RT-PCR; reverse transcription-polymerase chain reaction). In this case, the housekeeping gene RPL13A was used as the corrected reference mRNA.

The experimental results are shown in Figure 2a, Figure 2b and Figure 2c. The expression of keratin 1 and keratin 10 genes was reduced in the experimental group treated with external stimuli.

2) Protein analysis

Proteins were eluted by adding RIPA buffer (Radio-immunoprecipitation assay buffer) to the cultured cells. Next, the above-mentioned proteins were separated by SDS-PAGE, and keratin 1 and keratin 10 were confirmed and quantified by Western blot. In this case, glyceraldehyde-3-phosphate dehydrogenase (GAPDH; Glyceraldehyde 3-phosphate dehydrogenase) expressed from a housekeeping gene was used as a correction reference protein.

The experimental results showed that after the external stimulus treatment, the protein expression of keratin 1 and keratin 10 showed the same reduction phenomenon as the mRNA expression.

[Experimental example 3] Check whether psoriasis induces abnormal skin keratinization

Based on the results of Example 1 and Example 2 above, Experimental Example 3 was carried out in order to examine whether psoriasis would induce abnormal keratinization of the skin.

The implementation process of this embodiment 3 is the same as that of the above-mentioned embodiment 2, except that the external stimulus for treating the cultured keratinocytes in the above-mentioned embodiment 2 is replaced by adding 0.25 μg/ml, 1 μg/ml of psoriasis and then Culture was performed for 4 days.

The experimental results are shown in Figure 3a and Figure 3b. In the psoriasis-treated experimental group, the mRNA and protein expressions of keratin 1 and keratin 10 decreased.

Confirmation of psoriasis inhibitory effect of ginsenoside Rg1

Through the above-mentioned Examples 1 to 3, it has been confirmed that psoriasis is one of the inducing substances of abnormal keratinization of the skin. Next, an example for confirming the psoriasis inhibitory effect of ginsenoside Rg1 will be described.

[Example 1] Psoriasis inhibitory effect of ginsenoside Rg1-1

The stage of culturing skin keratinocytes was the same as that in Example 1 above. Next, the above-mentioned cells that were cultured were divided into two experimental groups, and then one of the experimental groups was treated with 1 μg/ml of a known psoriasis expression-inducing substance, that is, interleukin (IL-6; Interlukine-6). After treatment, another experimental group was treated with 1 μg of IL-6 and 10 ppm of ginsenoside Rg1 at the same time, and then cultured for 2 days. Among them, the above-mentioned ginsenoside Rg1 is a candidate substance for inhibiting the expression of psoriasis induced by IL-6.

After 2 days, the culture medium was removed, and mRNA analysis was carried out by the same method as the mRNA analysis in the above-mentioned Example 2, and then the difference in the mRNA expression level of psoriasis in the above two experimental groups was confirmed.

The experimental results are shown in Figure 4. Compared with the experimental group supplemented with IL-6 only, in the experimental group treated with IL-6 and ginsenoside Rg1 at the same time, the mRNA expression of psoriasis decreased.

[Example 2] Psoriasis inhibitory effect of ginsenoside Rg1-2 (in vitro)

The stage of culturing skin keratinocytes was the same as that in Example 1 above. Next, divide the cultured above-mentioned cells into two experimental groups, and then use 1 μg/ml of psoriasis to treat one of the experimental groups, and then use 1 μg/ml of psoriasis and 10 ppm of ginsenoside Rg1 to treat the other. One experimental group was treated and then cultured for another 4 days. After culturing, the mRNA and protein expression levels of keratin 1 and keratin 10 were confirmed according to the mRNA analysis and protein analysis methods in Example 2 above.

The experimental results are shown in Figure 5a and Figure 5b. As shown in Figure 5a and Figure 5b above, in the experimental group treated only with psoriasis, the expression levels of keratin 1 (KRT 1) and keratin 10 (KRT 10) were reduced, but the expression levels of keratin 1 (KRT 10) were reduced when using ginsenoside at the same time. In the experimental group treated with Rg1 and psoriasis, the mRNA expression levels of KRT 1 and KRT 10 were higher. In addition, in the experimental group treated with ginsenoside Rg1 and psoriasis at the same time, the mRNA expression level of KRT 1 was similar to that of the untreated experimental group, but the mRNA expression of KRT 10 was higher than that of the untreated experimental group. Group.

[Example 3] Psoriasis inhibitory effect of ginsenoside Rg1-3 (artificial skin)

In order to confirm the effect in an environment similar to the in vivo experiment, an experiment was conducted using artificial skin (MaTack).

The upper epidermis of one of the experimental groups was treated with 2 μg/ml of psoriasis, and the other experimental group was treated with 2 μg/ml of psoriasis and 10 ppm of ginsenoside Rg1, every 2 days within 1 week. A total of 3 treatments were performed each day. Next, the two experimental groups were fixed with formaldehyde solution and immunostained for keratin 1.

The dyed keratin 1 is red, and the observation results are shown in FIG. 6 . Compared with the untreated experimental group, in the experimental group treated only with psoriasis, the expression of keratin 1 was reduced, and the breakdown of the skin barrier led to loose tissue structure. But in the experimental group treated with psoriasis and ginsenoside Rg1 at the same time, the expression level of keratin 1 and the structure of the skin barrier returned to normal.

[Example 4]

Next, the compositions of the present invention shown in Table 1 were prepared based on the psoriasis inhibitory effect of the above-mentioned ginsenoside Rg1.

Table 1

*Numerical values of the comparative dosage form example and the implementation dosage form example represent the content thereof, and the unit is weight %.

The skin external preparations of comparative dosage form example 1 not containing ginsenoside Rg1 and implementation dosage form example 1 containing ginsenoside Rg1 at 0.1% by weight were produced, and 40 women aged 30 to 49 (4 experimental groups, 10 people in each group) As a subject, the above-mentioned skin external preparation was applied to the face every morning and evening for 8 weeks. Before the experiment (0 week), after 4 weeks, and after 8 weeks, the skin moisture content of each experimental group was measured three times, and the average comparison results of the above measurements are shown in Figure 7.

When measuring the moisture content in the skin, a skin moisture meter (Corneometer) capable of measuring the change value of the dielectric constant (dielectric constant) accompanying the moisture (hydration) of the stratum corneum of the skin is used.

The measurement results showed that the skin moisturizing effect when using the external skin preparation containing ginsenoside Rg1 was significantly higher than that when using the external skin preparation not containing ginsenoside Rg1.

Based on the above examples, it can be confirmed that the skin external preparation composition of the present invention is expected to provide skin protection and recovery effects for skin diseases such as atopic dermatitis and psoriasis.

Claims (8)

1. A skin external preparation composition, characterized in that: the active ingredient contains ginsenoside Rg1. 2. The skin external preparation composition according to claim 1, characterized in that: The structure of the ginsenoside Rg1 is shown in the following chemical formula 1: <chemical formula 1> 3. The skin external preparation composition according to claim 1 or 2, characterized in that: The content of the ginsenoside Rg1 is 0.001-5% by weight of the whole weight of the skin external preparation composition. 4. The skin external preparation composition according to claim 1, characterized in that: The ginsenoside Rg1 can inhibit the expression of psoriasis. 5. The skin external preparation composition according to claim 1, characterized in that: The skin external preparation composition is used for preventing or alleviating skin diseases. 6. The skin external preparation composition according to claim 1, characterized in that: The skin external preparation composition is used for moisturizing the skin. 7. The skin external preparation composition according to claim 5, characterized in that: The skin diseases include atopic dermatitis or psoriasis. 8. The skin external preparation composition according to claim 1, characterized in that: The skin external preparation composition includes at least one of a dermatological composition, a cleansing composition, and a cosmetic composition.