WO2023149766A1 - Cosmetic composition comprising saffron flower extract and danshen extract - Google Patents

Abstract

The present invention pertains to a cosmetic composition comprising a saffron flower extract and a Danshen extract. A mixture of the saffron flower extract and the Danshen extract has a synergistic effect on wrinkles, elasticity, and moisturization, and thus has an excellent effect on improving skin beauty.

Description

COSMETIC COMPOSITION COMPRISING Saffron Flower Extract and Danshen Root Extract

It relates to a cosmetic composition.

The skin is largely divided into three layers of tissues, epidermis, dermis, and hypodermis, in order from the outside, and functions to protect the human body from physical and chemical external environmental stimuli. Among them, the epidermis is further divided into the stratum corneum, the granular layer, the prickle layer, and the basal layer. The stratum corneum contains about 10 to 20% of water and exists in the outermost part of the human body, suppressing the evaporation of moisture to the outside of the body and excessive substances from the outside. It acts as a barrier to penetration. In addition, the surface of the stratum corneum is covered with a thin natural protective film made of sebum from the sebaceous glands and sweat from the sweat glands to prevent evaporation of moisture.

Wrinkles, reduced skin elasticity, skin sagging and dryness appearing in aged skin are mostly caused by changes in matrix proteins present in the dermis. The dermis plays a role in supporting the strength and shape of the skin within the skin, and as aging progresses, morphological changes occur in the dermis, which plays a decisive role in wrinkle formation and skin sagging. Collagen and Elastin play a role in supporting the dermal layer, and matrix metalloproteinase (MMP) is a representative enzyme that decomposes these matrix proteins. When irradiated with ultraviolet rays, the activity of MMPs in the skin increases, and collagen and other matrix proteins in the skin are significantly decomposed, which acts as a major factor in promoting skin aging.

Therefore, it is necessary to develop cosmetics with natural extracts that are excellent for preventing skin aging by promoting the synthesis of collagen and elastin and inhibiting MMP-1.

One aspect is to provide a cosmetic composition for improving skin conditions comprising a saffron flower extract and a Danshen root extract as active ingredients.

Another aspect is to provide a composition for external application for skin for improving skin conditions comprising a saffron flower extract and a Danshen root extract as active ingredients.

Another aspect is to provide a health functional food composition for improving skin conditions comprising a saffron flower extract and a Danshen root extract as active ingredients.

One aspect provides a composition for improving skin conditions comprising a saffron flower extract and a Danshen root extract as active ingredients.

The saffron ( Crocus Sativus ) is a dried spice prepared by extracting the pistil of a flower, and includes compounds such as crocin, picrocrocin, safranal, and crocetin. are doing Among the compounds, crocin is a glycoside mixture that gives saffron a yellow-red color, and the main component of crocin is α-crocin. Generally, saffron flowers are discarded after removing the stigma. In particular, saffron flowers, like pistils, have a deep red color due to the crocin component, so there is a risk of discoloration, so it is difficult to use them as cosmetic ingredients at high concentrations. In one embodiment, saffron flowers, specifically saffron petal extract, reduce the expression of MMP-1, which is increased by ultraviolet rays in skin fibroblasts, increase the expression of collagen, and promote cell migration in aged fibroblasts, thereby causing skin wounds. It was confirmed that not only can treat, but also exhibits a skin regeneration effect. Therefore, the saffron flower not only contains various active ingredients effective for improving skin beauty, but also has a lower cost compared to pistils, so it can be used as an alternative raw material.

Salvia Miltiorrhiza is a perennial herbaceous plant belonging to the Lamiaceae family, and is similar to ginseng in shape and is known to have a reddish color, promote blood circulation in the human body, and have excellent cholesterol-lowering effects.

The saffron flower extract is obtained by extracting saffron flowers with water, C1 to C4 lower alcohol, or a mixture thereof as a solvent, and the saffron flower extract may contain a crocin compound. After cutting the saffron flowers, it may be extracted by adding 2 to 20 times the weight of the saffron flowers with water, alcohol, or a mixture thereof. In addition, the extraction may be performed at 30 ° C to 100 ° C, and may be extracted for 1 to 10 hours. The extraction method may use cold needle extraction, ultrasonic extraction, or reflux cooling extraction method, and the number of extractions may be performed for 1 to 5 times.

In one embodiment, the saffron flower extract may be obtained by extracting the saffron flower with a mixture of water and a carbonhydrase as a solvent. In general, since carbohydrates have a high molecular weight and are insoluble, there is a problem in that they are difficult to absorb into the body. The carbolytic enzyme catalyzes a reaction that converts carbohydrates present in saffron flowers into monosaccharides such as glucose and maltose. Therefore, the saffron flower extract according to one embodiment contains a monosaccharide having a low molecular weight to promote skin absorption of active ingredients of the saffron flower, thereby maximizing the skin condition improvement effect.

In another embodiment, the saffron flower extract may be obtained by extracting saffron flowers with a mixture of water and a carbonhydrase as a solvent and decolorizing the saffron flowers by chemical, physical, electrical or biological methods. For example, the decolorization may be performed by a filtration method using a filtering agent. Since the crocin compound is removed by the decolorization, there is no risk of discoloration, so it can be used in cosmetics at a high concentration.

In one embodiment, regardless of the presence or absence of the crocin compound, skin wounds are reduced by reducing the expression of MMP-1, which is increased by ultraviolet rays in skin fibroblasts, increasing the expression of collagen, and promoting cell migration in aged fibroblasts. It was confirmed that not only can be treated, but also exhibits a skin regeneration effect. Therefore, the composition according to one embodiment may selectively apply a step of decolorizing the crocin compound according to the content of the saffron flower extract. In addition, the composition may have skin barrier strengthening, skin wrinkles, skin elasticity or moisturizing effects.

As used herein, the term "skin aging" refers to both tangible and intangible changes that appear in the skin with age, such as thinning of the epidermis, number of cells or blood vessels in the dermis, DNA damage repair ability, cell replacement cycle, It refers to reduction in wound healing, skin barrier function, epidermal moisture retention, sweat secretion, sebum secretion, vitamin D production, physical damage defense, chemical substance elimination ability, immune response, sensory function, and thermoregulation.

The composition may be used to improve skin aging caused by exogenous factors or endogenous factors. The extrinsic factor refers to various external factors, such as ultraviolet rays (light), and the endogenous factor is also referred to as a chronological factor and refers to a factor mainly caused by the passage of time. That is, the skin aging is not only premature aging symptoms induced by external stimuli such as ultraviolet rays, pollution, cigarette smoke, chemicals, etc., but also a natural aging phenomenon that occurs as the proliferation of skin cells decreases with age. It is a concept that includes wrinkles, loss of elasticity, skin sagging and dryness. In addition, wrinkles include stimulation caused by changes in internal and external factors that cause wrinkles by changing components constituting skin tissue.

The aging may be photoaging. The term "photoaging" is a phenomenon caused by external environmental factors, and the most representative factor is ultraviolet rays. Ultraviolet rays cause damage to biocomponents such as activation of proteolytic enzymes, chain scission of matrix proteins, and abnormal cross-linking, and repetition of these mechanisms results in apparent skin aging.

In this specification, the term "wrinkle" refers to a state in which elasticity of the skin is lost and loosened, and for example, the skin may be folded. The "prevention or improvement of skin wrinkles" may mean any action of preventing or improving wrinkles by suppressing the expression of wrinkle-related factors or increasing the total amount of collagen.

The "strengthen the skin barrier" may refer to any action that enhances the function of the skin barrier, which is located at the outermost layer of the skin and prevents loss of moisture and nutrients. Impaired skin barrier function may refer to any changes that occur in the skin due to reduced or damaged skin barrier function. Examples include increased skin wrinkling, dryness, dermatitis, atopic dermatitis, allergic dermatitis, acne, and the like.

The "moisturizing skin" may refer to any action that maintains skin moisture or prevents moisture loss.

The Danshen extract may be obtained by extracting Danshen with water, C1 to C4 lower alcohol, or a mixture thereof as a solvent. After cutting the Danshen root, it may be extracted by adding 2 to 20 times the weight of the Danshen root with water, alcohol, or a mixture thereof. In addition, the extraction may be performed at 30 ° C to 100 ° C, and may be extracted for 1 to 10 hours. The extraction method may use cold needle extraction, ultrasonic extraction, or reflux cooling extraction method, and the number of extractions may be performed for 1 to 5 times.

In one embodiment, the saffron flower extract and Danshen root extract may be mixed in a weight ratio of 1:0.01 to 1:10. The saffron flower extract and Danshen root extract are, for example, 1:0.01 to 1:10, 1:0.1 to 1:10, 1:1 to 1:10, 1:5 to 1:10, 1:5 to 1 :8 or 1:7 to 1:9. At this time, when the mixing ratio of the saffron flower extract and the Danshen root extract is less than the above range, there is a problem in that the synergistic effect of improving skin condition by the saffron flower extract and the Danshen root extract is not sufficiently exhibited.

In another embodiment, the composition is 0.001% to 80% by weight, for example, 0.01% to 60% by weight, 0.01% to 40% by weight, 0.01% to 30% by weight, based on the total weight of the composition, 0.01% to 20% by weight, 0.01% to 10% by weight, 0.01% to 5% by weight, 0.05% to 60% by weight, 0.05% to 40% by weight, 0.05% to 30% by weight, 0.05% by weight % to 20% by weight, 0.05% to 10% by weight, 0.05% to 5% by weight, 0.1% to 60% by weight, 0.1% to 40% by weight, 0.1% to 30% by weight, 0.1% to 0.1% by weight 20% by weight, 0.1% to 10% by weight, or 0.1% to 5% by weight of a mixture of saffron flower extract and Danshen root extract.

In one embodiment, it was confirmed that the mixture of the saffron flower extract and the Danshen root extract reduced the expression of MMP-1 increased by UV light and increased the expression of COL1A1, elastin and fibrillin, compared to each extract alone. In addition, it was confirmed that it exhibits a remarkable effect on the recovery of wound-induced fibroblasts. Therefore, the composition according to one aspect may have a synergistic activity of improving skin beauty and wound healing by the mixture of the saffron flower extract and the Danshen root extract.

The composition may be a cosmetic composition.

The cosmetic composition may have, for example, softening lotion, nutrient lotion, massage cream, nutrient cream, essence, pack, gel, ampoule, or skin-adhesive cosmetic formulation.

Ingredients included in the cosmetic composition may include ingredients commonly used in cosmetic compositions other than the composition as active ingredients, for example, conventional adjuvants and carriers such as stabilizers, solubilizers, vitamins, pigments and flavors. can include

In addition, the composition may be a composition for external application for skin.

The external skin preparation may be a cream, gel, ointment, skin emulsifier, skin suspension, transdermal delivery patch, drug-containing bandage, lotion, or a combination thereof. The external skin preparation is a component usually used in external preparations for skin such as cosmetics or pharmaceuticals, for example, water-based components, oil-based components, powder components, alcohols, moisturizers, thickeners, ultraviolet absorbers, whitening agents, preservatives, antioxidants, surfactants, and fragrances. , colorants, various skin nutrients, or combinations thereof and may be suitably blended as needed. The external skin preparations include metal sequestering agents such as disodium edetate, trisodium edetate, sodium citrate, sodium polyphosphate, sodium metaphosphate, and gluconic acid, caffeine, tannin, bellapamil, licorice extract, glabridin, and calin. Hot-water extracts of fruits, various herbal medicines, tocopherol acetate, glycyrrhizic acid, tranexamic acid and its derivatives or salts and other drugs, vitamin C, magnesium ascorbate phosphate, ascorbic acid glucoside, arbutin, kojic acid, glucose, fructose, Sugars, such as trehalose, etc. can also be mix|blended suitably.

In another embodiment, the composition may be a pharmaceutical composition.

The pharmaceutical composition may additionally include a pharmaceutically acceptable diluent or carrier. The diluent may be lactose, corn starch, soybean oil, microcrystalline cellulose, or mannitol, and magnesium stearate, talc, or a combination thereof as a lubricant. The carrier may be an excipient, a disintegrant, a binder, a lubricant, or a combination thereof. The excipient may be microcrystalline cellulose, lactose, low-substituted hydroxycellulose, or a combination thereof. The disintegrant may be calcium carboxymethylcellulose, sodium starch glycolate, calcium monohydrogen phosphate, or a combination thereof. The binder may be polyvinylpyrrolidone, low-substituted hydroxypropylcellulose, hydroxypropylcellulose, or a combination thereof. The lubricant may be magnesium stearate, silicon dioxide, talc, or a combination thereof.

The pharmaceutical composition may be formulated for oral or parenteral administration. Oral dosage forms may be granules, powders, solutions, tablets, capsules, dry syrups, or combinations thereof. Parenteral dosage forms may be injections.

The composition may be a health functional food composition.

The health functional food composition may be used alone or in combination with other foods or food ingredients, and may be appropriately used according to conventional methods. The mixing amount of the active ingredient may be appropriately determined depending on the purpose of use (prevention, health or therapeutic treatment). In general, when preparing food or beverage, the composition of the present specification may be added in an amount of 15 parts by weight or less based on the raw material. There is no particular limitation on the type of health functional food. Among the types of health functional foods, beverage compositions may contain various flavoring agents or natural carbohydrates as additional components, like conventional beverages. The natural carbohydrates include monosaccharides such as glucose and fructose, disaccharides such as maltose and sucrose, polysaccharides such as dextrin and cyclodextrin, and sugar alcohols such as xylitol, sorbitol, and erythritol. As the sweetener, natural sweeteners such as thaumatin and stevia extract, or synthetic sweeteners such as saccharin and aspartame may be used. The health food composition may also contain nutrients, vitamins, electrolytes, flavors, colorants, pectic acid and its salts, alginic acid and its salts, organic acids, protective colloidal thickeners, pH adjusters, stabilizers, preservatives, glycerin, alcohol, and carbonated beverages. carbonation agent used, or a combination thereof. The health functional food composition may also contain natural fruit juice, fruit juice beverages, fruit flesh for preparing vegetable beverages, or a combination thereof.

Another aspect also provides a method of preventing, ameliorating, or treating a condition in a subject comprising treating or administering to a subject in need thereof an effective amount of a composition described above.

The condition of the subject may be a skin related condition.

As used herein, the terms "administering," "introducing," and "implanting" are used interchangeably and according to one embodiment into a subject by a method or route that results in at least partial localization of a composition to a desired site. It may mean the arrangement of the composition according to one embodiment of.

Administration may be administered by a method known in the art. Administration can be administered directly to a subject by any means, for example, by routes such as intravenous, intramuscular, oral, transdermal, mucosal, intranasal, intratracheal or subcutaneous administration. can The administration may be administered systemically or locally.

The subject may be a mammal, such as a human, cow, horse, pig, dog, sheep, goat, or cat. The subject may be a subject in need of skin condition improvement, for example, skin moisturizing, skin barrier strengthening, skin wound treatment, and skin wrinkle improvement.

The administration is 0.1 mg to 1,000 mg per day of the composition according to one embodiment, for example, 0.1 mg to 500 mg, 0.1 mg to 100 mg, 0.1 mg to 50 mg, 0.1 mg to 25 mg, 1 mg to 1,000 mg, 1 mg to 500 mg, 1 mg to 100 mg, 1 mg to 50 mg, 1 mg to 25 mg, 5 mg to 1,000 mg, 5 mg to 500 mg, 5 mg to 100 mg, 5 mg to 50 mg , 5 mg to 25 mg, 10 mg to 1,000 mg, 10 mg to 500 mg, 10 mg to 100 mg, 10 mg to 50 mg, or 10 mg to 25 mg may be administered. However, the dosage may be variously prescribed depending on factors such as formulation method, administration method, patient's age, weight, sex, medical condition, food, administration time, administration route, excretion rate and response sensitivity, and those skilled in the art can Dosage can be appropriately adjusted in consideration of these factors. The number of administrations can be once a day or two or more times within the range of clinically acceptable side effects, and administration can be done at one or two or more sites, daily or at intervals of 2 to 5 days, for a total of The number of administration days may be administered from 1 day to 30 days per treatment. If necessary, the same treatment can be repeated after a titration period. For non-human animals, the same dosage per kg as for humans is used, or the above dosage is converted by the volume ratio (eg, average value) of the organ (heart, etc.) between the target animal and the human. A single dose can be administered.

The cosmetic composition according to one aspect contains both a saffron flower extract and a Danshen root extract, and thus has a synergistic effect on wrinkles, elasticity, and moisturizing compared to each single extract, and thus has an excellent effect on improving skin conditions.

1 is a photograph showing the color of a saffron flower extract with or without a decolorization process.

Figure 2 is a graph showing the effect of saffron flower extracts on the expression of MMP-1 according to the presence or absence of a crocin compound.

Figure 3 is a graph showing the effect of saffron flower extract on collagen production according to the presence or absence of a crocin compound.

Figure 4 is a result showing the effect of saffron flower extract on wound healing and skin regeneration according to the presence or absence of a crocin compound.

Figure 5 is a graph showing the effect of the cosmetic composition according to one embodiment on the expression of MMP-1.

Figure 6 is a graph showing the effect of the cosmetic composition according to one embodiment on the expression of COL1A1.

7 is a graph showing the effect of the cosmetic composition according to one embodiment on the production of elastin.

8 is a graph showing the effect of the cosmetic composition according to one embodiment on the expression of fibrillin.

9A and 9B are results showing the skin regeneration promoting effect of the cosmetic composition according to one embodiment.

10A and 10B are graphs showing the skin moisturizing effect of the cosmetic composition according to one embodiment.

11A, 11B and 11C are results showing the skin wrinkle improvement effect according to the formulation of the cosmetic composition according to one embodiment.

Hereinafter, a preferred embodiment is presented to aid understanding of the present invention. However, the following examples are provided to more easily understand the present invention, and the content of the present invention is not limited by the following examples.

[Production Example]

Preparation Example 1. Preparation of saffron flower water extract (1)

1 L of water and carbonhydrase were added to 16.7 g of dried saffron flowers, followed by reflux cooling and extraction at 25° C. for 24 hours. This was filtered using filter paper (Advantec no. 2 filter paper), and then the filtrate was concentrated under reduced pressure using a vacuum concentrator (EYELA Co., N-1000, Japan) to prepare a saffron flower water extract.

Preparation Example 2. Preparation of saffron flower water extract (2)

The saffron flower dried extract prepared in Preparation Example 1 further included a step of decolorizing the colored material using a filtering agent to which 0.5 to 10% by weight of activated carbon was added based on the total weight of the solvent for 10 to 24 hours. A saffron flower water extract was prepared in the same manner as in Preparation Example 1 except for the above.

Manufacturing Examples 3 to 5. Preparation of saffron flower water extract (3)

To 12.0 g of dried saffron flowers, 1 L of water at various temperatures (25° C., 50° C., and 80° C.) was added, followed by reflux cooling and extraction for 24 hours. This was filtered using filter paper (Advantec no. 2 filter paper), and then the filtrate was concentrated under reduced pressure using a vacuum concentrator (EYELA Co., N-1000, Japan) to prepare a saffron flower water extract.

Manufacturing Examples 6 to 7. Preparation of saffron flower water extract (4)

Saffron flower dried extract water extract was prepared in the same manner as in Preparation Example 2, except that the saffron flower water extract prepared in Preparation Examples 3 and 5 was used.

Production Examples 8 to 10. Preparation of Saffron Flower Alcohol Extract (5)

1 L of butylene glycol of various concentrations (25%, 50%, and 75%) was added to 10 g of dried saffron flowers, followed by reflux cooling and extraction at 50° C. for 24 hours. This was filtered using filter paper (Advantec no. 2 filter paper), and then the filtrate was concentrated under reduced pressure using a vacuum concentrator (EYELA, N-1000, Japan) to prepare a dried saffron flower butylene glycol extract. .

Production Examples 11 to 13. Preparation of Saffron Flower Alcohol Extract (6)

Saffron flower ethanol extracts were prepared in the same manner as in Preparation Example 8, except that 25 ° C. ethanol (10%, 25% and 50%) was used.

Preparation Example 14. Preparation of Danshen root extract

1 L of 25% butylene glycol was added to 100 g of Danshen root powder, followed by reflux cooling and extraction at 25° C. for 24 hours. This was filtered using a filter paper (Advantec no. 2 filter paper), and then the filtrate was concentrated under reduced pressure using a vacuum concentrator (EYELA, N-1000, Japan) to prepare a Danshen root extract.

[Example]

Examples 1-3. Preparation of Cosmetic Composition Containing Saffron Flower Extract and Danshen Root Extract

A cosmetic composition was prepared by mixing the saffron flower water extract prepared in Preparation Example 2 and the Danshen root extract prepared in Preparation Example 14 at weight ratios of 1:0.1, 1:0.5, and 1:1.

Example 4. Preparation of a Creamy Cosmetic Composition Containing Saffron Flower Extract and Danshen Root Extract

A cosmetic composition in a cream formulation containing the ingredients and contents of Table 1 below was prepared. In this specification, unless otherwise indicated, content is % by weight.

ingredient	content
Purified water	52.3625
sodium phytate	0.02
Hydroxyacetophenone	0.2
glycerin	5
panthenol	One
pentylene glycol	2
1,2-Hexanediol	One
glycerin	2
xanthan gum	0.07
tromethamine	0.15
sodium carbomer	0.35
Hydroxyethyl Acrylate/Sodium Acryloyl Dimethyl Taurate Copolymer	0.0475
Polyglyceryl-3methylglucose distearate	2
Polyglyceryl-3-distearate	1.89
Glyceryl Stearate Citrate	0.11
Caprylic/Capric/Myristic/Stearic Triglyceride	0.5
coconut palm oil	2
squalane	2
Caprylic/Capric Triglyceride	4
Purified water	20
Acrylates/C10-30 Alkyl Acrylate Crosspolymer	0.2
Purified water	2.94
Preparation Example 2	0.03
1,2-Hexanediol	0.015
caprylyl glycol	0.015
Butylene Glycol	0.085
Purified water	0.012
Preparation Example 14	0.003

[Comparative example]

Comparative Example 1. Preparation of a creamy cosmetic composition

A creamy cosmetic composition was prepared in the same manner as in Example 4, except that the components and contents of Table 2 were included.

ingredient	content
Purified water	55.4625
sodium phytate	0.02
Hydroxyacetophenone	0.2
glycerin	5
panthenol	One
pentylene glycol	2
1,2-Hexanediol	One
glycerin	2
xanthan gum	0.07
tromethamine	0.15
sodium carbomer	0.35
Hydroxyethyl Acrylate/Sodium Acryloyl Dimethyl Taurate Copolymer	0.0475
Polyglyceryl-3methylglucose distearate	2
Polyglyceryl-3-distearate	1.89
Glyceryl Stearate Citrate	0.11
Caprylic/Capric/Myristic/Stearic Triglyceride	0.5
coconut palm oil	2
squalane	2
Caprylic/Capric Triglyceride	4
Purified water	20
Acrylates/C10-30 Alkyl Acrylate Crosspolymer	0.2

[Experimental Example]

Experimental Example 1. Identification of color and crocin compounds of saffron flower extracts according to decolorization process

The surface color of the saffron flower extract prepared in Preparation Examples 1 to 13 according to the decolorization process was visually observed. In addition, the content of the crocin compound corresponding to the main component of the saffron flower extract was confirmed by HPTLC (High-Performance Thin Layer Chromatography) and shown in Table 3.

Preparation Example 1

Preparation Example 2

Preparation Example 3

Production Example 4

Preparation Example 5

Preparation Example 6

Preparation Example 7

Preparation Example 8

Preparation Example 9

Preparation Example 10

Preparation Example 11

Preparation Example 12

Preparation Example 13

crocin concentration

+/-

-

+/-

+/-

+/-

-

-

++++

++

+/-

+/-

++

++++

++++: Large amount detected

++: detected

+/-: small amount detected

-: not detected

1 is a photograph showing the color of a saffron flower extract with or without a decolorization process.

As a result, as shown in FIG. 1, it was confirmed that Preparation Example 9, which did not include a bleaching process, had a red color, but Preparation Example 2, which additionally included a bleaching process, exhibited a yellow tint.

In addition, as shown in Table 1, when the bleaching process was not included, the crocin compound was detected at a certain concentration or higher, but in Preparation Examples 2 and 6-7, which further included a bleaching process, the crocin compound was not detected.

That is, it can be seen that the crocin compound, which is a chemical substance responsible for the color of saffron, is removed by decolorization.

Experimental Example 2. Confirmation of anti-aging and anti-wrinkle activity according to the presence or absence of crocin compound

Saffron contains compounds such as crocin, picrocrocin, and safranal as main components. Therefore, it was confirmed whether a difference in anti-aging and anti-wrinkle activity occurs depending on the presence or absence of the crocin compound, which is one of the main components. Specifically, a human dermal fibroblast cell line (Human dermal fibroblast, Hs68) was cultured for 48 hours in an incubator at 37°C and 5% CO ₂ conditions. Then, UVA 10 J/cm ² was irradiated or not irradiated through a Biosun UV lamp. Immediately after UVA irradiation, Preparation Example 2 and Preparation Example 9 were treated by concentration (1.0% and 2.0%), and further cultured for 48 hours. Then, the expression level of MMP-1 was measured by ELISA. As a positive control group, 10 μg/mL or 100 μg/mL of Crocin compound was used.

Figure 2 is a graph showing the effect of saffron flower extracts on the expression of MMP-1 according to the presence or absence of a crocin compound.

As a result, as shown in Figure 2, it can be confirmed that the expression of MMP-1 was increased in the fibroblasts by irradiation of ultraviolet rays, and the increased expression of MMP-1 was reduced by treating Preparation Examples 2 and 9. there was.

Experimental Example 3. Confirmation of skin elasticity improvement effect according to the presence or absence of crocin compound

The effect of improving skin elasticity according to the presence or absence of the crocin compound was confirmed. Specifically, a human dermal fibroblast cell line (Human dermal fibroblast, Hs68) was cultured for 48 hours in an incubator at 37°C and 5% CO ₂ conditions. Thereafter, Preparation Example 2 and Preparation Example 9 were treated by concentration (1.0% and 2.0%), and further cultured for 24 hours. Thereafter, the culture supernatant was stored at -80 ° C, and the amount of collagen expression was measured by ELISA. As a positive control group, 10 μg/mL of Crocin compound was used.

Figure 3 is a graph showing the effect of saffron flower extract on collagen production according to the presence or absence of a crocin compound.

As a result, as shown in Figure 3, Preparation Example 2 and Preparation Example 9 was confirmed to increase the expression of collagen in a concentration-dependent manner. In particular, in the case of Preparation Example 2 in which the Crocin compound was removed by decolorization, it was confirmed that collagen expression was significantly increased compared to Preparation Example 9 containing the Crocin compound. Therefore, the saffron flower extract according to one embodiment may exhibit an effect of improving skin elasticity regardless of the presence or absence of the crocin compound.

Experimental Example 4. Confirmation of skin wound healing and regeneration effects according to the presence or absence of crocin compound

Skin wound healing and regeneration effects according to the presence or absence of the Crocin compound were confirmed. Specifically, a human dermal fibroblast cell line (Hs68) was cultured in an incubator at 37° C. and 5% CO ₂ to reach 100% confluency. Thereafter, using a pipette tip, randomly scrape across the center of the well to make a wound, and then replace the above Preparation Example 2 and Preparation Example 9 with a medium containing 1.0%, respectively, for an additional 24 hours. cultured. Then, cell migration of the fibroblasts was observed using an electron microscope. As a positive control group, 10 μg/mL of Crocin compound was used.

Figure 4 is a result showing the effect of saffron flower extract on wound healing and skin regeneration according to the presence or absence of a crocin compound.

As a result, as shown in Figure 4, fibrotic cells aged by wounding significantly reduced cell migration by young cells, and by treating senescent cells with the Crocin compound, Preparation Example 2 and Preparation Example 9 It was confirmed that cell migration was significantly increased. In particular, in the case of Preparation Example 2, cell migration was increased by 38% or more compared to senescent cells, showing a remarkable cell migration effect compared to the Crocin compound and Preparation Example 9, in which cell migration increased by 8%.

Therefore, the saffron flower extract according to one embodiment can not only treat skin wounds by promoting cell migration in aged fibroblasts, but also exhibit skin regeneration effects, even though it does not contain a crocin compound.

Experimental Example 5. Confirmation of anti-aging and anti-wrinkle synergistic activity

In view of the results of the above experimental examples, it can be confirmed that the saffron flower extract has a skin condition improvement effect regardless of the presence or absence of the crocin compound. Therefore, in the following, it was confirmed whether the mixture of the saffron flower extract and the Danshen root extract has synergistic activity of anti-aging and anti-wrinkle compared to each single extract. Specifically, human dermal fibroblast cell line (Human dermal fibroblast, Hs68) was divided into 3x10 ⁵ in a 6-well plate, and then cultured for 24 hours in an incubator at 37°C and 5% CO ₂ conditions. Thereafter, the medium was removed, DPBS was added, and 20 mJ/cm ² of UVB was irradiated or not irradiated, and Examples 1 to 3 were added at a concentration of 1%, respectively, and further cultured for 24 hours. Thereafter, RNA was isolated from the cells of each sample using Trizol (RNA iso, DAKARA, Japan), RNA was quantified at 260 nm with nanodrop, and cDNA was synthesized in an amplifier using 2 μg of RNA. (C1000 Thermal Cycler, Bio-Rad, USA). Using the synthesized cDNA as a template, cybergreen (SYBR Green supermix, Applied Biosystems, USA) was added together with primers and cDNA for the target genes MMP-1 (Matrix metalloproteinase-1) and COL1A1, and real-time PCR machine (Step One Plus, Applied Biosystems, USA) real-time polymerase chain reaction was performed. The expression level of the gene was finally analyzed through correction for the β-actin gene. EGCG 1 μM was used as a positive control.

Figure 5 is a graph showing the effect of the cosmetic composition according to one embodiment on the expression of MMP-1.

Figure 6 is a graph showing the effect of the cosmetic composition according to one embodiment on the expression of COL1A1.

As a result, as shown in FIG. 5, it was confirmed that the expression of MMP-1 was increased in the fibroblasts by UV irradiation, and the increased expression of MMP-1 was decreased by treating Examples 1 to 3. Specifically, it was confirmed that Examples 1 to 3 significantly reduced the expression of MMP-1 compared to the case of treatment with the saffron flower extract alone. In addition, as shown in FIG. 6, it was confirmed that Examples 1 to 3 significantly increased COL1A1 expression compared to Preparation Examples 2 and 14.

That is, the cosmetic composition according to one aspect reduces the expression of MMP-1 increased by ultraviolet rays and increases the expression of COL1A1 by including a saffron flower extract and a Danshen root extract, thereby having excellent anti-aging and anti-wrinkle activity. can

Experimental Example 6. Confirmation of synergistic effect of improving skin elasticity

It was confirmed whether the mixture of the saffron flower extract and the Danshen root extract had a synergistic activity of improving skin elasticity compared to each single extract. Specifically, the human dermal fibroblast cell line (Human dermal fibroblast, Hs68) was inoculated in a 6-well plate at a density of 4X10 ⁵ and then cultured at 37°C, 5% CO ₂ incubator for 24 hours. Thereafter, after washing with PBS, each of Examples 1 to 3 was treated at a concentration of 1% in DMEM medium without FBS, and further cultured for 24 hours. Thereafter, the gene expression levels of COL1A1, elastin (ELN), and fibrillin (FBN) were measured in the same manner as above, except that primers for COL1A1, elastin (ELN), and fibrillin (FBN) were used. analyzed.

7 is a graph showing the effect of the cosmetic composition according to one embodiment on the production of elastin.

8 is a graph showing the effect of the cosmetic composition according to one embodiment on the expression of fibrillin.

As a result, as shown in FIGS. 7 and 8 , it was confirmed that Examples 1 to 3 significantly increased the expression of elastin and fibrillin compared to Preparation Examples 2 and 14.

That is, since the cosmetic composition according to one aspect includes a saffron flower extract and a Danshen root extract, the expression of elastin and fibrillin is significantly increased compared to each single extract, and thus, a synergistic effect of improving skin elasticity can be exhibited.

Experimental Example 7. Confirmation of skin wound healing and regeneration synergistic effect

It was confirmed whether the mixture of the saffron flower extract and the Danshen root extract had a synergistic activity of skin wound healing and regeneration compared to each single extract. Specifically, the human dermal fibroblast cell line (Human dermal fibroblast, Hs68) was dispensed into a 6-well plate at a concentration of 5x10 ⁵ cells/well, and then cultured for 24 hours in an incubator at 37°C and 5% CO ₂ conditions to achieve 100% confluency. It was made into a state of confluency. Thereafter, after arbitrarily scratching the center of the well using a pipette tip, the medium was replaced with a medium containing 1.0% of Examples 1 to 3, respectively, and further cultured for 24 hours. . Then, cell migration of the fibroblasts was observed using an electron microscope. As a positive control group, bEGF 10 ng/ml was used.

9A and 9B are results showing the skin regeneration promoting effect of the cosmetic composition according to one embodiment.

As a result, as shown in FIGS. 9A and 9B, it was confirmed that Examples 1 to 3 showed a remarkable effect on the recovery of scarred fibroblasts compared to Preparation Example 14.

That is, the cosmetic composition according to one aspect may have excellent effects on wound healing and skin regeneration by including the saffron flower extract and the Danshen root extract.

Experimental Example 8. Confirmation of skin moisturizing and wrinkle improvement effects

It was confirmed whether the mixture of the saffron flower extract and the Danshen root extract had effects of moisturizing skin and improving wrinkles. Specifically, 5 subjects (average age 45 years old) were asked to use Example 4 and Comparative Example 1 twice a day for 4 weeks on the left/right side of the face. Then, the test site was washed with soap and waited in a constant temperature and humidity room (22 ± 2 ℃, 40 ~ 60% RH) for 10 minutes, and then the skin moisture level (0 weeks) was measured and skin wrinkles (0 weeks) were photographed. . After 2 weeks and 4 weeks of product use, skin moisture level and skin wrinkles were measured in the same way, and the average of the three measured values in each week was calculated and compared with the measured values in the 0th week.

10A and 10B are graphs showing the skin moisturizing effect of the cosmetic composition according to one embodiment.

11A, 11B and 11C are results showing the skin wrinkle improvement effect of the cosmetic composition according to one embodiment.

As a result, as shown in FIGS. 10A and 10B, it was confirmed that the skin moisture level after 4 weeks of use was 24.3% and 5.0% in Example 4 and Comparative Example 1, respectively, and increased compared to before use. In particular, in Example 4, it was confirmed that the skin moisture level was significantly increased compared to Comparative Example 1.

In addition, as shown in FIGS. 11A to 11C, it was confirmed that Example 4 and Comparative Example 1 decreased skin indentation as the use time increased. In particular, in Example 4, it was confirmed that the rate of change in skin subsidence at the 4th week of use was significantly reduced compared to Comparative Example 1.

That is, the cosmetic composition according to one aspect has an excellent effect on skin moisturizing and wrinkle improvement.

The above description of the present invention is for illustrative purposes, and those skilled in the art can understand that it can be easily modified into other specific forms without changing the technical spirit or essential features of the present invention. will be. Therefore, the embodiments described above should be understood as illustrative in all respects and not limiting.

Claims (8)

1. A cosmetic composition for improving skin condition comprising saffron flower extract and Danshen root extract as active ingredients.
2. The cosmetic composition according to claim 1, wherein the saffron flower extract is obtained by extracting water, C1 to C4 lower alcohol, or a mixture thereof using a solvent.
3. The method according to claim 1, wherein the saffron flower extract is extracted with a mixture of water and carbohydrate decomposition enzyme (carbonhydrase) as a solvent; and

   A cosmetic composition obtained by decolorizing the saffron flower extract.
4. The cosmetic composition according to claim 1, wherein the Danshen root extract is obtained by extracting water, C1 to C4 lower alcohol, or a mixture thereof with a solvent.
5. The cosmetic composition according to claim 1, wherein the skin condition improvement is skin aging, skin barrier strengthening, wound healing, skin wrinkles, skin elasticity or moisturizing.
6. The cosmetic composition according to claim 1, wherein the saffron flower extract and the Danshen root extract are mixed in a weight ratio of 1:0.01 to 1:10.
7. A composition for external application for skin for improving skin conditions, comprising a saffron flower extract and a Danshen root extract as active ingredients.
8. A health functional food composition for improving skin conditions comprising saffron flower extract and Danshen root extract as active ingredients.

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