KR101155714B1 - Cosmetic composition containing immature fruit or seed extract of torreya nucifera - Google Patents

Abstract

The present invention relates to a cosmetic composition containing an immature fruit or seed extract of a non-tree, more specifically, by containing an immature fruit or seed extract of a non-tree as an active ingredient, antioxidant, anti-aging, whitening or wrinkle improvement effect It relates to an excellent cosmetic composition.

Description

Cosmetic composition containing immature fruit or seed extract of non-mature fruit

The present invention relates to a cosmetic composition containing an immature fruit or seed extract of a non-tree, more specifically, by containing an immature fruit or seed extract of a non-tree as an active ingredient, antioxidant, anti-aging, whitening or wrinkle improvement effect It relates to an excellent cosmetic composition.

The jasmine tree is an evergreen tree of the creeper's cultivars of attention. nucifera , 25 m high, 2 m in diameter. The branches spread all over, the bark is greyish brown, and on the old tree they are shallowly split off. Leaves are 25 mm long, 3 mm wide, hard in strips, pointed at end, arranged in rows like feathers. The leaf surface is dark green, the back is brown and the middle is only on the back. Petioles of about 3 mm long and fall after 6-7 years.

The flowers are single flowers and bloom in April. A male flower has about 10 bracts, brown and about 10 mm long, with 10 flowers hanging on a peduncle. The female flower is an irregularly shaped egg-shaped, which runs two or three in one place and is wrapped in five or six green cloths. Fruits ripen in September-October the following year and are elliptical, 25-28 mm long, 20 mm in diameter, and 3 mm thick.

Seeds are oval, 23 mm long, 12 mm in diameter, dark brown, and hard. Only seeds are called visas and are medicinal.

Lost water is a tree grown for the purpose of fruit production, and fruit is an important forest product. Fruit maturation is the process by which fruits become edible and, in general, they become softer. The study of the physical, chemical and physiological changes that fruit matures and changes is important in determining the fruit's production, storage, transport and harvest time.

Free radicals produced by various physical, chemical and environmental factors such as enzymes, reducing metabolism, chemicals, pollutants and photochemical reactions in the body are non-selective and irreversible destruction of cell components such as lipids, proteins, sugars and DNA. It is known to cause various diseases including cell aging or cancer through the action. In addition, various peroxides in the body, including lipid peroxides produced as a result of lipid peroxidation by these free radicals, may cause oxidative destruction of cells and cause various functional disorders.

Accordingly, antioxidants such as free radical scavengers or peroxide production inhibitors are expected as agents for inhibiting or treating aging and various diseases caused by these oxides.

On the other hand, skin aging can be classified into two types according to the factors. One of them is natural aging, which causes the skin's structure and physiological function to continually decline with age, while the other, aging, is caused by accumulated external stress such as sunlight. It is. In particular, ultraviolet rays (UV) are one of the well-known causes of aging, and the skin exposed to ultraviolet rays for a long time causes the stratum corneum to become thicker, and collagen and elastin, which are the main components of the skin, are deteriorated, thereby losing the elasticity of the skin. These collagen and elastin are regulated by various factors. Collagen and elastin produced by the expression of matrix metalloprotease such as collagenase and elastase are degraded, resulting in skin The collagen content in the inside is reduced.

Various substances have been developed and used for the purpose of suppressing the reduction of collagen and elastin, which causes the decrease in elasticity. Among them, retinoids such as retinol and retinoic acid show elasticity improving effect (Dermatology therapy, 1998, 16, 357-364), protein fractions obtained from Leguminosae Seeds have also been applied to show an elasticity-boosting effect. However, these retinoids have a disadvantage in that irritation occurs even when only a small amount is applied to the skin.

On the other hand, a number of factors are involved in determining the skin color of the human, including the activity of melanocytes (melanocytes) that make melanin pigment, the distribution of blood vessels, the thickness of the skin and the presence or absence of pigments, such as carotenoids, bilirubin, etc. Factors are important.

Especially, the most important factor is a black pigment called melanin which is produced by the action of various enzymes such as tyrosinase in melanocytes in the human body. The formation of this melanin pigment is influenced by genetic factors, physiological factors related to hormone secretion, stress, and environmental factors such as ultraviolet irradiation.

Melanin pigment, which is produced from melanocytes of the skin of the body, is a phenolic polymer material having a complex form of black pigment and protein. It protects skin organs below the dermis by blocking ultraviolet rays from the sun and free radicals generated in the skin body. It acts to protect the proteins and genes in the skin.

As such, melanin produced by stress stimulation inside and outside the skin is a stable substance that does not disappear until it is released to the outside through skin keratinization even if the stress disappears. However, if more melanin is produced than necessary, it causes hyperpigmentation such as blemishes, freckles, and spots, resulting in cosmetically bad results.

In addition, as the leisure population increases, the number of people who enjoy being active outside increases the demand to prevent melanin pigmentation caused by ultraviolet rays.

In response to such demands, ascorbic acid, kojic acid, arbutin, hydroquinone, glutathione or derivatives thereof, or substances having tyrosinase inhibitory activity have been used in combination with cosmetics or medicines. Its use is limited due to insufficient whitening effect, safety problems on skin, formulation and stability problems when formulated in cosmetics, etc.

Accordingly, the present inventors have searched for several natural products as a part of the research to find an effective substance for antioxidant, anti-aging, whitening and wrinkle improvement while solving the above problems, antioxidant, in the cosmetics containing immature non-tree fruit extract, It was found that the anti-aging, whitening and wrinkle improvement effect is more excellent to complete the present invention.

Accordingly, it is an object of the present invention to provide a cosmetic composition excellent in antioxidant, anti-aging, whitening and anti-wrinkle effect.

In order to achieve the above object, the present invention provides a cosmetic composition containing an immature fruit or seed extract of the non-tree as an active ingredient.

In the present invention, the cosmetic composition containing the immature fruit or seed extract of the non-tree has anti-aging effect through promoting collagen biosynthesis, suppressing the expression of elastase and collagenase, along with antioxidant effects resulting from DPPH and ROS production inhibitory effects. It was confirmed that the whitening effect due to the effect of inhibiting melanin production and improving pigmentation produced by ultraviolet light could be exhibited. Therefore, the immature fruit or seed extract of the non-noodle or radish according to the present invention can be very useful as a cosmetic composition or pharmaceutical composition for antioxidant, anti-aging, whitening, wrinkle improvement.

The present invention relates to a cosmetic composition containing an immature fruit or seed extract of non-tree as an active ingredient.

The immature fruits or seeds of the non-visa tree used in the present invention were collected between April and September before the visa was fully matured, and more preferably from June to September.

In the present invention, the fruit extract extracted from the immature fruit taken from the non-tree with water or an organic solvent can be used. In the present invention, after separating the flesh and seeds from the immature fruit collected from the non-tree, it can be used alone or mixed with the extract extracted with water or organic solvent, respectively.

The organic solvent used in the present invention may be selected from the group consisting of ethanol, methanol, butanol, hexane, ether, ethyl acetate, chloroform and a mixed solvent of these organic solvents and water, and preferably ethanol is used. At this time, the extraction temperature is preferably 10 ~ 80 ℃, can be extracted for 6 to 24 hours. If the extraction temperature and extraction time is out of the extraction efficiency may decrease or change of components may occur.

The cosmetic composition according to the present invention preferably contains from 0.01 to 10% by weight of the immature fruit or seed extract of the non-tree relative to the total weight of the composition. At this time, when the content of the non-extract extract is less than 0.01% by weight, it is difficult to achieve the required efficacy, when the content of more than 10% by weight does not show more efficacy.

Cosmetic composition according to the present invention is an antioxidant effect due to the DPPH and ROS production inhibitory effect; Anti-aging and anti-wrinkle effect through promoting collagen biosynthesis, suppressing expression of elastase and collagenase; And a whitening effect due to the effect of inhibiting melanin production and improving pigmentation produced by ultraviolet light.

Hereinafter, the content of the present invention will be described in more detail through examples and test examples. These examples are provided only for understanding the contents of the present invention, and the scope of the present invention is not limited to these examples, and modifications, substitutions, and insertions commonly known in the art may be performed. This is also included in the scope of the present invention.

[Example 1] Preparation of non-fruit extract by harvesting time

In order to prepare non-berry extracts by harvesting time, immature fruit-1 (April to May), immature fruit-2 (April to July), immature fruit-3 (August to September), and ripening fruit (10 Mon-November) fruits were collected and dried, and 6 liters of hexane was added to 500 g of each fruit. After stirring and extracting three times at room temperature, 4 liters of 80% ethanol was added to 1 kg of skim berries. Reflux extraction was followed by immersion at 15 ° C. for 1 day. Then, the residue and the filtrate were separated by filter cloth filtration and centrifugation, and the separated filtrate was concentrated under reduced pressure to obtain each extract.

[Test Example 1] Change in total polyphenol content of non-berry fruit extracts by harvest time

Folin-Ciocalteu's method was used to determine the change in total polyphenol content of non-berry fruit at each harvest time. That is, 8.4 mL of distilled water and 0.5 mL of 2 N Folin-Ciocalteu's reagent (Sigma) were added to 0.1 mL of each extract prepared in Example 1, and 1 mL of 10% Na ₂ CO ₃ was added thereto, followed by 1 hour of reaction. Absorbance was measured at 760 nm and then the amount was converted to a standard curve using catechin. The results are shown in Table 1 below.

Changes in Total Polyphenol Contents at Different Ripening Seasons Test substance Total Polyphenol Content (%) Immature and -1 1.5 Immature 3.9 Immature and -3 4.6 Ripening 3.1

As a result of analyzing the total polyphenol content of the extract of each time of maturity of the non-berry fruit in Table 1, the total polyphenol content of the immature fruit (immature fruit-2 and immature fruit-3) extract collected in June to September is mature More than the extract was confirmed.

Test Example 2 Antioxidant Effect of Non-berry Fruit Extracts by Harvest Time (DPPH Test)

The antioxidant activity was evaluated by the change in absorbance generated by the reduction of the organic radical DPPH (1,1-diphenyl-2-picryl hydrazyl) (an antioxidant is oxidized). The degree of inhibition of DPPH oxidation by each test substance was measured by measuring the change in absorbance which is decreased compared to the untreated group, and the concentration showing an absorbance of 50% or less compared to that of the control group was evaluated as the effective antioxidant concentration.

190 μl of 100 μM (dissolved in ethanol) DPPH solution and 10 μl each of the non-fruit extract and the positive control group prepared in Example 1 were prepared, followed by reaction for 30 minutes at 37 ° C. and absorbance at 540 nm. Was measured. As a positive control group, a widely used synthetic antioxidant Trolox was used.

DPPH analysis results (% inhibition) of each material are shown in Table 2 below.

IC ₅₀ : Sample concentration when absorbance was reduced by 50% by the added sample Test substance IC ₅₀ (ppm) Immature and -1 1250 Immature 952 Immature and -3 80 Ripening 160 Trolox 65

In the results of Table 2, the immature fruit extract according to the present invention showed a better antioxidant activity than the ripe fruit and extracts, it was confirmed that showing a similar level of antioxidant capacity as trolox, a positive control.

Test Example 3 Inhibitory Activity of Reactive Oxygen Species Formation of Non-rip Fruit Extracts by Harvest Time

Human keratinocytes HaCaT cell line (Dubeccos Modification of Eagles Medium, FBS 10%) added with penicillin / streptomycin and dispensed 2.0 × 10 ⁴ per well into 96 well black plate for fluorescence measurement Using 37 ° C., 5% CO ₂ Incubated for 1 day under conditions. To this, penicillin / streptomycin-added serum-free DMEM (no FBS) was added to the test substance described in Table 3 at a concentration of 10 ^-4 molar concentration of 37 ℃, 5% CO ₂ After incubation for 24 hours in a culture medium for 1 day, washed with HCSS (HEPES-buffered control salt solution) to remove the remaining medium and then DCFH-DA (2 ', 7'-dichlorodihydro) prepared in HCSS 20 μM 100 μl of fluorescein diacetate, Molecular Probes, Inc) was added at 37 ° C., 5% CO _2. Incubated for 20 minutes under the conditions and washed with HCSS. Then, 100 μl of the treated HCSS was added to each sample concentration, and then the fluorescence intensity of DCF (dichlorofluorescein) oxidized to ROS was measured by a fluorescence plate reader (Ex = 485 nm, Em = 530 nm). Thereafter, UVB (30 mJ / cm ² ) was irradiated, and the fluorescence intensity immediately after the treatment and 3 hours after the treatment was measured by a fluorescence plate reader (Ex = 485 nm, Em = 530 nm). Trolox was used as a positive control.

Results of the test for inhibiting ROS production of each test substance are shown in Table 3 below.

ROS production inhibitory test results (% of untreated group) Concentration (ppm) Immature and -1 Immature Immature and -3 Ripening Trolox 500 64.1 43.8 36.6 53.2 58.5 250 79.7 54.8 46.6 61.7 73.3 100 88.6 61.4 52.0 71.5 74.6 50 96.3 75.6 63.2 88.2 76.9

In the results of Table 3, the extracts of immature fruit-2 and 3 showed better ROS production inhibitory activity than the ripe fruit and extracts, and in particular, it was confirmed that the ROS production inhibitory ability was more excellent than that of the positive control trolox.

[Test Example 4] Collagen Biosynthesis Promoting Effect of Non-fruit Fruit Extracts by Harvest Time

The collagen biosynthesis-promoting effect of the non-berry fruit extract of each sampling time of Example 1 was measured in comparison with tocopherol and EGCG.

First, fibroblasts (PromoCell, Germany) were seeded (10 ⁵ per well) in a 24-well plate and cultured to 90% growth. This was incubated with serum-free DMEM medium for 24 hours, and then treated with the non-berry fruit extract of Example 1 and the positive control group (tocopherol and EGCG) dissolved in serum-free medium at 10 ^-4 molarity, respectively, and CO ₂ incubator for 24 hours. Incubated at. These supernatants were removed and procollagen increased or decreased using a procollagen type (I) ELISA kit. The results are shown in Table 4 below, in which the collagen synthesis ability is compared to the non-treated group as 100.

Test substance Collagen Synthesis (%) Untreated group 100 Tocopherol 118 EGCG 125 Ripening 102 Immature and -3 145 Immature 138 Immature and -1 115

In the results of Table 4, it was confirmed that the immature fruit extract according to the present invention increased the amount of collagen biosynthesis than the ripe fruit extract, it was confirmed that the collagen biosynthesis ability is superior to the positive control tocopherol and EGCG.

[Test Example 5] Collagenase expression inhibitory effect of the extract of non-berry fruit by the harvesting time

The collagenase expression inhibitory effect (producing ability) of the non-fruit fruit extract of Example 1 was measured as follows in comparison with tocopherol and EGCG.

First, human fibroblasts were put into 5,000 cells / well in a 96-well microtiter plate containing DMEM medium containing 2.5% fetal bovine serum, and cultured to 90% growth. It was. After 24 hours of incubation in serum-free DMEM medium, the non-berry fruit extract of Example 1 and the positive control group (tocopherol and EGCG) dissolved in serum-free DMEM medium as test substance were treated for 10 hours at 10 ^-4 molarity. After that, the cell culture was collected.

Thereafter, the collected cell culture solution was measured for the degree of collagenase production using a commercially available collagenase measuring instrument (Amersham Pharma Co., Ltd., USA).

First, the collected cell culture solution was put in a 96-well plate uniformly coated with primary collagenase antibody, and the antigen-antibody reaction was performed in a thermostat for 3 hours. After 3 hours, the chromophore-conjugated secondary collagen antibody was placed in a 96-well plate and reacted again for 15 minutes. After 15 minutes, a color-causing substance was added to induce color development at room temperature for 15 minutes, and 1 M sulfuric acid was added to stop the reaction (color development). .

The absorbance of the yellow-colored 96-well plate was measured at 405 nm using an absorbance meter, and the expression level of collagenase was calculated by Equation 1 below. At this time, the reaction absorbance of the collected cell culture solution of the group not treated with the test substance was taken as the absorbance of the untreated group without the test substance.

Collagenase expression level (%) = A / B × 100

A: absorbance of the test substance treatment cell group

B: absorbance of untreated group

On the other hand, the results of measuring the collagenase expression inhibitory effect of the test substances in the human fibroblasts are shown in Table 5 below.

Test substance Collagenase expression level (%) Untreated group 100 Tocopherol 75 EGCG 60 Ripening 98 Immature and -3 64 Immature 66 Immature and -1 83

In the results of Table 5, it was confirmed that the immature fruit extract according to the present invention inhibits collagenase expression more effectively than the ripe fruit extract.

[Test Example 6] Inhibitory effect of elastase expression of non-berry fruit extracts by collection time

Elastase expression inhibitory effect (producing ability) of the non-fruit extract of the above Example was measured as follows in comparison with tocopherol and EGCG.

First, human fibroblasts were placed in a 96-well microtiter plate containing DMEM medium containing 2.5% fetal bovine serum to 5,000 cells / well, and cultured until 90% growth. After 24 hours of incubation with serum-free medium, the non-fruit extract and the positive control group (tocopherol and EGCG) of the above example dissolved in serum-free medium as test substance were treated at 10 ^-4 molarity for 24 hours, and then the cell culture solution. Was taken.

Thereafter, the amount of elastase production was measured using the commercially available elastase measuring instrument (Amersham Pharmacia, USA).

First, the cell culture medium was placed in a 96-well plate uniformly coated with the primary elastase antibody, and the antigen-antibody reaction was performed in a thermostat for 3 hours. After 3 hours, the chromophore-bound secondary elastin antibody was placed in a 96-well plate and reacted again for 15 minutes. After 15 minutes, the coloring stimulant was added to induce color development at room temperature for 15 minutes, and 1M sulfuric acid was added to stop the reaction (color development).

The absorbance of the yellowish 96 well plate was measured at 405 nm using an absorbance meter, and the expression level of elastase was calculated by the following Equation 2. At this time, the reaction absorbance of the collected cell culture solution of the group not treated with the test substance was taken as the absorbance of the untreated group.

Elastase expression level (%) = A / B × 100

A: absorbance of the test substance treatment cell group

B: absorbance of untreated group

On the other hand, the results of measuring the efficacy of inhibiting the expression of the elastase of the test substance in the human fibroblasts are shown in Table 6, which is prepared by the degree of expression of the elastase of the untreated group as 100.

Test substance Elastase expression level (%) Untreated group 100 Tocopherol 88 EGCG 68 Ripening 87 Immature and -3 56 Immature 63 Immature and -1 72

In the results of Table 6, it was confirmed that the immature fruit extract according to the present invention more effectively suppress the expression of elastase than the ripe fruit extract.

[Test Example 7] melanin production inhibitory effect of non-berry fruit extracts by collection time

To determine the melanin production inhibitory effect of the extract of the non-fruit fruit of Example 1 was measured using the pigment cells of the rat.

First, mouse Mel-Ab cells derived from C57BL / 6 mice (Dooley, TP et al , Skin pharmacol, 7, pp 188-200) were placed in DMEM with 10% fetal placental serum, 100 nM 2-O- Tetracanocanoyphorbol-13-acetate and 1 nM cholera toxin were incubated at 37 ° C. and 5% CO ₂ . Cultured Mel-Ab cells were detached with 0.25% trypsin-EDTA, and the cells were cultured at a concentration of 10 ⁵ cells / well in a 24 well plate, and the non-berry fruit extract and positive control of Example 1 were consecutive for 3 days from the second day. Phosphorus hydroquinone was added at 10 ppm and incubated. Then, the culture medium was removed, washed with PBS, cells were dissolved with 1 N sodium hydroxide, absorbance was measured at 400 nm, and the melanin production inhibition rate was calculated according to the following Equation 3, and the results are shown in Table 7 Dooley's way).

Test substance Melanin production inhibition rate (%) Untreated group 100.0 Ripening 84.5 Immature and -3 38.6 Immature 38.2 Immature and -1 56.7 Hydroquinone 41.1

In the results of Table 7, it was confirmed that the immature fruit extract according to the present invention was superior to the melanin production inhibitory effect than the mature fruit extract, in particular showing a melanin production inhibition rate similar to the hydroquinone positive control group.

Claims (7)

delete delete delete Antioxidant cosmetic composition containing a pulp extract of immature fruit of a non-tree taken in June to September as an active ingredient. An anti-aging cosmetic composition containing a pulp extract of immature fruit of a non-tree taken in June to September as an active ingredient. A whitening cosmetic composition containing a pulp extract of immature fruit of a non-tree taken in June to September as an active ingredient. A cosmetic composition for improving wrinkles, comprising a pulp extract of immature fruit of a non-tree obtained in June to September.