KR101081424B1 - External Skin Composition Containing Extract of Medicinal Plants - Google Patents

Abstract

The present invention relates to an external preparation for skin containing at least one extract selected from anemarrhenae Rhizoma, Polygalae Radix, Cynomorii Herba, and Evodiae Fructus, which is an insulin-like growth factor-1. like Growth Factor-1; IGF-1) promotes in vivo secretion of various factors related to skin aging to improve or reduce skin wrinkles and elasticity, which ultimately delays skin aging. to provide.

Hair, base, sessile, sewage, IGF-1, growth promotion, skin aging prevention, wrinkle improvement, elasticity improvement

Description

External Skin Composition Containing Extract of Medicinal Plants             

1 is a graph of the prolidase activity enhancement of each extract.

2 is a western blot for prolyl-4-hydroxylase of each extract.

3 is a graph of type I procollagen enhancement of each extract.

Figure 4 is a replica analysis of the results of the wrinkles of the wrinkles when applying the hairless mice of each extract.

5 is a replica analysis result graph for wrinkle improvement when applying the mixture of each hairless mouse.

The present invention relates to a composition for promoting the secretion of insulin-like growth factor-1 (hereinafter referred to as "IGF-1") in vivo, and more specifically, conventionally used as a herbal medicine The present invention relates to an external skin composition for promoting IGF-1 in vivo secretion by containing at least one extract of Anemarrhenae Rhizoma, Polygalae Radix, Cynomorii Herba, Evodiae Fructus.

Growth hormone (GH), which has been widely used in the clinic since the 1950s, is a hormone secreted and regulated by the pituitary gland and hypothalamus. IGF-1 is known to not only control the secretion of growth hormone in vivo, but is also closely linked to growth hormone, and plays an important role in cell growth, differentiation and induction of activity.

IGF-1 is known to help form bones, muscles, nerves, blood vessels and tissues, help regenerate damaged cells and inhibit apoptosis. In addition, it is known to delay aging of the pituitary gland, normalize blood sugar stability and insulin resistance, and be involved in the growth and differentiation of cells and tissues by being involved in glucose metabolism, protein metabolism, and fat metabolism (DL Roith. Et al ., Endocrine Reviews 22 (1): 53 (2001)). In vivo IGF-1 is produced primarily by the liver but has also been reported to be secreted through autocrine / paracrine in body tissues as well (D'Ercole AJ. Et al ., Dev. Biol 1980; 75: 315) This suggests that growth hormone and IGF-1 may be closely related, but each may respond independently. In addition, IGF-1 produced in the liver is involved in controlling the feedback loop with growth hormone, and IGF-1, which is secreted locally in each tissue, has been reported to induce cell growth and differentiation. This could be seen in the liver specific igf-1 gene-deleted mouse model, which significantly reduces the amount of IGF-1 that is secreted from the liver and systemically circulated due to the lack of that gene. Despite this, tissue growth and differentiation were normal (sjogren K. et al., Proc Natl Acad Sci 1999; USA 96: 7088). This suggests that even without systemic circulation, localized growth of IGF-1 can promote cell growth and activity.

Meanwhile, various studies have been conducted on the relationship between IGF-1 and skin. IGF-1, which plays an important role in cell growth, differentiation and activator induction, plays an important role in skin cell growth by promoting the cell cycle, and collagen and fibronectin, which are extracellular matrix (ECM) components in the dermis. Is known to increase the expression of TGFβ-1, which plays an important role in GAG synthesis (Ghahary A et al , Journal of Cellular Physiology 1998; 174: 301-309) and inhibit the expression of MMP enzymes that degrade ECM components in the dermis. (Ghahary A et al , Growth Factors, 2000; 17 (3): 167-76). In addition, it is known to promote the expression of lysyl oxidase, lysyl hydroxylase and collagen recycling, which are important for collagen fiber structure formation (Reiser K et al , Am J Physiol, 1996; 271 (3pt2): R696-703).

As described above, it can be seen that IGF-1, which has various and important functions in vivo, is secreted and exhibits its function in connection with growth hormone or through its own mechanism. In addition, the secretion of IGF-1 independent of growth hormone is important to the growth of each tissue through direct or indirect function of mediating the action of other activators. Therefore, it is important to secure a substance having IGF-1 secretion ability that is safe and effective for the human body in terms of growth promotion and aging inhibition.

The present inventors continued to study and select safe natural substances in vivo while promoting the secretion of IGF-1 in vivo, and as a result, extracts obtained from conventionally used herbal medicines such as hair, raw paper, stiletto, and filthy milk were extracted from in vivo IGF-1. In vitro studies have confirmed that it promotes secretion, and it was also confirmed that each extract having these characteristics showed the effect of anti-aging. And as an example of the composition comprising each extract and applied to the animal model to the external preparation of the skin as a result of strengthening the ECM components in the skin to confirm that the wrinkle formation and suppression is increased to complete the present invention.

In the present invention, it is intended to provide a hair, a base, a stiletto, sewage extract having an effect of promoting the secretion of IGF-1.

In addition, the present invention is to provide a hair, raw paper, scalpel, sewage extract having an effect of delaying the aging of the skin.

The present invention relates to a composition for promoting the secretion of insulin-like growth factor-1 (hereinafter referred to as "IGF-1") in vivo, and more specifically, conventionally used as a herbal medicine The present invention relates to an external skin composition for promoting IGF-1 in vivo secretion by containing at least one extract of Anemarrhenae Rhizoma, Polygalae Radix, Cynomorii Herba, Evodiae Fructus.                     

Anemarrhenae Rhizoma used in the present invention is a subterranean stem of Anemarrhena asphodeloides Bunge, a liliaceae plant, acting on the lungs, stomach, and kidneys to lower the heat and moisturize the lack of essence. , Sedation, antibacterial, hypoglycemic is known.

Polygalae Radix is the root of Polygala tenuifolia Willd., Which is applied to the treatment of nervous breakdown, heart system, forgetfulness, sea water, insomnia, etc. Anti-inflammatory action is known.

Cynomorii Herba is a fleshy stem of the herbaceous and perennial herbaceous herbaceous herbaceous cyst ( Cynomorium songaricum Rupr.), Which is known to improve oil and yang deficiency, weakness in the lower back and knee, and to be effective in menstrual irregularities and constipation. .

Evodiae Fructus is an immature fruit of rhyme and plant erectile oil ( Evodia rutaecarpa Benth.). It is used in expectoration, dry stomach, pain , hypertension, eczema, and pharmacological action is known as antibacterial, central excitement, and uterine contraction.

As a result of examining the growth and skin aging-related patents for the substance, in the case of Jimo, Korean Patent Publication No. 99-0080969 describes the contents of 'soluble soluble extract having growth hormone secretion stimulating activity and growth hormone secretion factor isolated therefrom'. In the case of the original paper, Korean Patent Publication No. 2000-0038805 has a patent as a growth accelerator in the form of a composition with other substances. . This can be said that the approach is different from the aging inhibitory and growth promotion implemented through the direct expression of IGF-1 in the skin and changes in skin aging-related factors through the present invention. Therefore, the present invention can be meaningful to reveal a new skin aging inhibitory function through the direct growth factor production promotion and to show the potential as a growth accelerator.

In another aspect, the present invention provides a skin external preparation composition for promoting IGF-1 secretion by delaying skin aging by containing one or more of the hair, base paper, yangyang, sewage extract as an active ingredient.

The hair, base, sesame and sesame oil extract according to the present invention are contained in an amount of 0.001 to 10% by weight, preferably 0.01 to 5% by weight, based on the total weight of the external preparation for skin, based on in vitro and in vivo experiments. It is good to be.

Formulation form of the external preparation for skin according to the present invention includes a cosmetic composition having a formulation of a flexible cosmetics, nourishing cosmetics, massage creams, nutrition creams, packs, gels or skin adhesive type cosmetics, and also lotions, ointments, gels, creams Formulations of transdermal dosage forms such as, patches or sprays. In addition, in the external preparation composition of each formulation, other components than the essential components described above can be appropriately selected and blended by those skilled in the art without difficulty according to the formulation or purpose of use of other external preparations. The composition is used for preventing or improving skin wrinkles.

In the present invention, the extract is dried, pulverized any one or more selected from jimo, raw paper, yangyang, sewage and extracted by heating under reflux in 95% ethanol, filtering using a filter paper after cooling, filtered filtrate after filtration Extraction is carried out through evaporation using a reduced pressure evaporator.

Hereinafter, the structure and the effect of the present invention will be described in more detail with reference to Examples and Test Examples. However, the present invention is not limited only to these examples.

Example 1

Preparation of Hair, Raw Grass, Ambush and Sewage Extract

After drying and grinding 1kg each of gimo, paper, sediment, and sewage oil, 95% ethanol was added at a weight (Kg) ratio of 4 times and then heated at 80 ° C. for 2 hours to extract the active ingredient. This was allowed to stand at room temperature for 1 day to precipitate impurities, and the filtrate was filtered twice using a filter paper and concentrated by completely evaporating using a reduced pressure evaporator.

[Test Example 1]

Enhancement of IGF-1 Promoter Activity

Dr. After transfection of HEK 293 cells with plasmid containing IGF-1 promoter donated by Peter Rotein (Washington University of Medicine), IGF-1 promoter activity was observed by reporter gene assay. 10 ⁵ cells were added to a 24-well plate and incubated in DMEM medium containing 10% fetal bovine serum for 24 hours, and each extract prepared in Example 1 was treated at a concentration of 1 ppm and 10 ppm. After 24 hours, washed with PBS, cells were disrupted to obtain a supernatant. Subsequently, luciferase activity of the negative control group not treated with the extract only in the same composition as the sample treated with the extract prepared in Example 1 using the Dual-Luciferase ^R Reporter Assay System kit manufactured by Promega was measured. It is shown in Table 1 below. As a result, each extract was confirmed to have higher IGF-1 promoter activity than the negative control group.

TABLE 1

[Test Example 2]

IGF-1 mRNA expression level using RT-PCR

Human fibroblasts were placed in 100 mm-Dish at a concentration of 10 ⁵ and then cultured in DMEM medium containing 10% fetal calf serum, and the four extracts were replaced with a medium containing 10 ppm. After 8 hours of culture, total RNA was obtained from the cells, and then cDNA was synthesized using a first strand cDNA synthesis kit (MBI Fermentas), followed by 94 ° C 1min, 57 using a human IGF-1 primer (Forward-GACAGGGGCTTTTATTTCAAC, Reverse-CTCCAGCCTCCTTAGATCAC). 35 cycles were carried out under the reaction conditions of 1 min at 72 ° C. and 1 min at 72 ° C. to obtain a human IGF-1 band. The results are shown in Table 2. As shown in Table 2, it was found that the relative amount of IGF-1 mRNA expression was increased compared to the negative control group, and through this result, each extract has an ability to promote IGF-1 secretion.

TABLE 2

[Test Example 3]

Enhancement of Prolidase Activity

Human fibroblasts were placed in 100 mm-dish at a concentration of 10 ⁶ and incubated at 37 ° C. in DMEM medium containing 10% fetal bovine serum, and each extract prepared in Example 1 was replaced with a medium containing 1 ppm and 10 ppm. . After 24 hours of culture, the cells were obtained and the change of enzyme activity of each test group was measured according to the prolidase activity measurement method of Myara (Myara et al, Clin. Chim. Acta 125, 1982), and compared with the negative control group. Indicated. Samples treated with each extract prepared in Example 1 showed high enzymatic activity as compared to the negative control. Prolidase is known to be an enzyme that acts as a reproducing function of proline, an amino acid that is important for collagen synthesis in the cell. Therefore, the increase of glycoenzyme activity is known to be closely related to the increase in collagen synthesis.

[Test Example 4]

increased production of prolyl-4-hydroxylase (P4H)

Human fibroblasts were placed in 100 mm-dish at a concentration of 10 ⁶ and incubated at 37 ° C. in DMEM medium containing 10% fetal bovine serum, and each extract prepared in Example 1 was replaced with a medium containing 1 ppm and 10 ppm. . After 48 hours of culture, cells were disrupted to obtain proteins, followed by electrophoresis, and western blot was performed using an antibody specific for P4H. The results are shown in FIG. 2. As a result of the experiment, the expression level was increased compared to the control group treated with the vehicle, and each extract was considered to influence the increase of P4H expression. Since P4H attaches hydroxyl groups to proline residues of procollagen in the cell, it acts as a rate of collagen production rate that is involved in the formation of correct tertiary structure. Therefore, increased production of sugar enzymes may contribute to skin wrinkle improvement.

[Test Example 5]

Measurement of Collagen Biosynthesis Efficacy of Skin Cells

Human fibroblasts were placed in a 24-well plate at a concentration of 10 ⁴ and then cultured in DMEM medium containing 10% fetal bovine serum, and then each extract prepared in Example 1 was replaced with a medium containing 1 ppm and 10 ppm. After 48 hours of culture, the supernatant was harvested, and the amount of procollagen produced was quantified using the ELISA (Takara MK101) method. As a result, a comparison value of the control 100 was shown in FIG. 3. As a result, it was found that each extract prepared in Example 1 increased type I collagen biosynthesis. It seems that each extract is effective in improving skin aging.

[Test Example 6]

Inhibition of Biosynthesis of Type I Collagenase (MMP-1) by UV Light

Human fibroblasts were placed in a 24-well plate at a concentration of 10 ⁴ , incubated at 37 ° C. in DMEM medium containing 10% fetal bovine serum, and each extract prepared in Example 1 was replaced with a medium containing 1 ppm and 10 ppm. It was. After 48 hours of culture, the supernatant was harvested to quantify the amount of type I collagenase produced using ELISA (AP biotech RPN2610) method. As a result, it can be seen that each extract prepared in Example 1 reduces the biosynthesis of type I collagenase by UV rays, which can effectively prevent the decomposition of skin substrates and contribute to skin wrinkle improvement. .

[Table 3]

[Test Example 7]

Confirmation of wrinkle improvement effect using animal model

In order to determine the skin wrinkle improvement effect of the extracts of Example 1 in the skin, the application test was carried out using about 7 weeks old hairless mouse (SKH-1 female hairless mouse). Cream formulations 1 to 4 containing each extract prepared as shown in Table 4 for use in the test were prepared and used as a sample, and the test group used as a control group did not contain any of the extracts. Example 1 was made and treated at the same dose. The content of each component is weight percent.                     

[Table 4]

As shown in Table 4, test materials were prepared and applied to the back of the hairless mice for 11 weeks with UV irradiation. Ultraviolet irradiation was carried out every other day, and the material was applied every day. Afterwards, the effect of wrinkle improvement was confirmed by replica wrinkle analysis. As can be seen in Figure 4, the analysis results showed that all four extracts have an anti-wrinkle effect.

In addition, after making the cream formulation in the form of a mixture of four extracts as shown in Table 5, the effect of wrinkle improvement was confirmed by the same test method using the hairless mice. Ultraviolet irradiation, material application and analysis were performed in the same way. As can be seen in Figure 5 it can be seen that the wrinkle improvement effect was the best when all four extracts were applied.

TABLE 5

As described above, according to the results of Test Examples 2 to 7, the hair, raw paper, yangyang, sewage extract prepared in the present invention can delay skin aging by effectively controlling various indicator factors related to skin aging. Confirmed.

Therefore, the composition for external application of the skin containing the hair, raw paper, stiletto and sesame oil extracts according to the present invention increases the expression of IGF-1 in cells, which is associated with the activity of prolidase and prolyl-4-hydroxylase, which are important for the production of collagen. Increasing production promotes collagen synthesis and ultimately contributes to skin aging.

As described above, the hair, base paper, stingray, sesame oil extract and external skin composition according to the present invention promoted the in vivo secretion of IGF-1. In addition, the external preparation composition for skin used in the present invention contains one or more active ingredients of hair, raw paper, yangyang and sesame oil extracts known as herbal medicines, thus having safety to the human body and improving or inhibiting various factors related to skin aging to improve wrinkles and It is thought to exhibit an elasticity improvement effect and ultimately delay skin aging. In addition, each extract according to the present invention can be used to prepare a composition having one or more as an active ingredient with anti-aging and growth promoting effect.

Claims (6)

A skin external preparation cosmetic composition characterized by promoting the secretion of insulin-like growth factor (IGF-1) containing sessile sheep, sessile oil or a mixed extract of both. The method of claim 1, The extract for external application cosmetic composition, characterized in that contained in 0.001 to 10% by weight relative to the total weight of the composition. The method of claim 1, The composition for external application cosmetic composition for skin wrinkle prevention or improvement. The method of claim 1, The extract is a cosmetic composition for external application to the skin, characterized in that the mixed extract of the hair, base paper or both. The method of claim 1, The extract is (a) dry grinding the amount of sewage, sewage oil or a mixture of the two and extracting by heating under reflux in 95% ethanol; (b) filtering the extract in step (a) using a filter paper after cooling; And (c) completely evaporating the filtrate filtered in step (b) using a reduced pressure evaporator; Skin external preparation cosmetic composition characterized in that it is extracted. The method of claim 5, The extract is a cosmetic composition for external application to the skin, characterized in that the mixed extract of the hair, base paper or both.

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