KR20220012211A - The cosmetic composition for improving acne skin containing cork oak extract - Google Patents

Abstract

Provided is a cosmetic composition for alleviating acne skin. The cosmetic composition for alleviating acne skin contains a Quercus suber bark extract, a Quercus root extract, a Thymus vulgaris extract, coconut oil, and camelina seed oil. According to the present invention, provided is the cosmetic composition having a stable formulation that is not only excellent in alleviating acne skin but also hypoallergenic to the skin through the cosmetic composition containing a Quercus suber extract.

Description

The cosmetic composition for improving acne skin containing cork oak extract

The present invention relates to a cosmetic composition for improving acne prone skin, and more particularly, to a cosmetic composition for improving acne prone skin comprising a cork oak extract.

Acne refers to an inflammatory skin disease of the hair follicles, hair follicles and surrounding tissues caused by the action of hormones, excessive sebum secretion, and bacterial infection.

There are several causes of acne. The first cause is excessive secretion of sebum. Sebum is a substance secreted by the sebaceous glands, and in a normal amount, it prevents harmful substances from the outside of the skin from penetrating between the keratinocytes. However, under circumstances such as puberty, before and after menstruation, early pregnancy, taking contraceptives, and stress, the human body exhibits a different hormonal balance from the normal state. This changed hormonal balance stimulates the sebaceous glands, which leads to excessive secretion of sebum. Also, there are cases of genetically high sebum secretion, and sebum secretion can also be increased by a change in external temperature or an increase in skin temperature due to wrong eating habits.

The second cause is an infection of the pores. The excessively secreted sebum contains a lot of fatty acids, which raises the pH of the skin surface and dissolves the skin's lipids, thereby lowering the skin's own defense power. On the surface of the skin where the pH is increased, the bacteria that cause acne easily proliferate, and the proliferated acne bacteria causes purulent inflammation through the weakened skin.

Regarding hormones among the causes of acne, acne is activated by male hormones, and male hormones increased during puberty enlarge the sebaceous glands (skin oil glands) of the skin. There are many sebaceous glands on the face, back, and chest areas where acne often occurs. The sebaceous glands are connected to ducts containing hair called hair follicles, and in these sebaceous glands an oily substance called sebum is produced. This sebum is discharged to the skin surface through the opening of the hair follicle. Sebum stimulates the inner wall of the hair follicle to cause the cells to fall out more quickly, and the cells that have fallen out are agglomerated and block the pores of the hair follicle to form a comedone, the basic lesion of acne. Propionibacterium acnes (P. acnes), a bacterium residing in hair follicles, secretes a lipolytic enzyme. This enzyme breaks down triglycerides in sebum to form free fatty acids, which in turn stimulates hair follicles to burst the hair follicle wall. Eventually, sebum, bacteria, and exfoliated cells are released into the skin, causing erythema, swelling, pus, or pimples. The immunological response to Propionibacterium acnes contributes to the inflammatory response of acne. It is a well-known fact that there is a family history of acne. However, its exact genetic pattern is still unclear.

Propionibacterium acnes is a bacterial bacterium that feeds on sebum and causes trouble as it grows. In normal people, it is normally present on the skin and does not cause any problems. fly After eating fat (triglyceride), the fungus excretes a substance similar to butter, that is, fatty acid. These chemical changes cause the pH balance of the oil on the skin surface to be broken, and the weak acidity of healthy skin to become alkalized, eventually causing the skin's own function to deteriorate and a vicious cycle of the skin.

In order to prevent and alleviate acne caused by Propionibacterium acnes, treatment is mainly using topical or oral medications. Topically applied antibiotics (clindamycin, erythromycin), which have a direct sterilization effect on acne bacteria, and prevent the formation of free fatty acids, are drugs made by modifying vitamin A. There are skin regeneration ointments (tretinoin, adaptalene) that exfoliate dead skin cells and make it easier to drain sebum, and acne ointment (benzoyl peroxide) that reduces bacterial population with a strong antibacterial agent and has a slight anti-inflammatory and comedonal dissolution action. . There are two main types of oral medications: antibiotics that sterilize acne bacteria and reduce inflammation, and retinoids made by modifying vitamin A with a drug commonly known as roaccutane. Retinoid is well known as a sebum control agent because it reduces the secretion of sebum, but this drug not only controls sebum but also causes other causes of acne, such as abnormal keratinization (abnormal keratinization showing incomplete and immature keratinization), inflammatory reaction, and bacterial growth. It helps to block all pathways of acne outbreaks.

These oral drug regimens use retinoids and adrenocortical hormones for the purpose of suppressing the function of the sebaceous glands and the destruction of the hair follicle wall. It can be, so be careful when using it. Therefore, research on substances that can have such a medical treatment effect at the stage of cosmetics before reaching the stage of treating acne within the medical range is in progress, but there are not many such substances. In particular, cosmetics can be used for a long time compared to therapeutic drugs and can cause less damage to normal skin, so it is necessary to develop cosmetic raw materials that are safe for the skin and have excellent acne improvement effects from natural materials.

Korean Patent Registration 10-1652244 (Announcement Date August 30, 2016)

The technical problem to be solved by the present invention is an antibacterial effect against acne-causing bacteria as well as an effect of regulating the secretion of sebum essential for improving acne, and having a whitening effect, containing a cork oak extract that can prevent skin discoloration To provide a cosmetic composition for improving acne skin.

Another technical problem to be solved by the present invention is to provide a cosmetic composition for improving acne prone skin comprising a cork oak extract having the effect of improving acne skin in a wide range of stages.

However, the technical problems to be solved by the present invention are not limited to the above problems, and may be variously expanded without departing from the technical spirit and scope of the present invention.

A cosmetic composition for improving acne prone skin according to an embodiment of the present invention for solving the above problems is characterized in that it comprises a cork oak bark extract, an oak root extract, a thyme extract, coconut oil and camelina seed oil.

According to an embodiment of the present invention, the cork oak bark extract contains 2% by weight, the oak root extract contains 1% by weight, the thyme extract contains 0.3% by weight, and the coconut oil contains 0.5% by weight %, and the camelina seed oil is characterized in that it comprises 0.2% by weight.

According to an embodiment of the present invention, the thyme extract is characterized in that it includes substances extracted from flowers, leaves and stems of thyme.

According to an embodiment of the present invention, the cork oak bark extract, the oak root extract and the thyme extract include: a) nitrogen freezing and freezing in an environment of -80° C. or less by including moisture in the plant to be extracted; b ) grinding the frozen plant to form a powder, c) mixing the frozen powder with vegetable oil so that the effective component of the plant to be extracted is included in the oil, d) ultrasonic and high-frequency components in the step c) Including the steps of accelerating the extraction of effective components of plants by oscillating toward the extraction liquid phase, and e) when the extraction process is completed, it undergoes a filtration process to finally produce a plant extract in an oil-based liquid state. characterized by completion.

Other specific details of the invention are included in the detailed description and drawings.

According to the present invention, a cosmetic composition comprising a cork oak extract is not only excellent in improving acne skin but also hypoallergenic and has a stable formulation.

In addition, according to the present invention, it is possible to not only improve acne skin, but also suppress skin pigmentation through a whitening effect.

However, the effects of the present invention are not limited to the above effects, and may be variously expanded without departing from the spirit and scope of the present invention.

1 is a clinical trial result data using a cosmetic composition according to Example 1 of the present invention.
2 is a clinical trial result data using the cosmetic composition according to Example 2 of the present invention.
3 is a result data of tyrosinase activity inhibition experiment using Arbutin.
4 is a result data of the tyrosinase activity inhibition experiment using Example 1 of the present invention.
5 is a picture of the solution before the tyrosinase activity inhibition experiment using Example 1 of the present invention.
6 is a photograph showing the color change of the solution after the tyrosinase activity inhibition experiment using Example 1 of the present invention.

Advantages and features of the present invention and methods of achieving them will become apparent with reference to the embodiments described below in detail in conjunction with the accompanying drawings. However, the present invention is not limited to the embodiments disclosed below, but will be embodied in various different forms, and only these embodiments allow the disclosure of the present invention to be complete, and common knowledge in the art to which the present invention pertains It is provided to fully inform those who have the scope of the invention, and the present invention is only defined by the scope of the claims.

The terminology used herein is for the purpose of describing the embodiments, and is not intended to limit the present invention. In this specification, the singular also includes the plural unless specifically stated otherwise in the phrase. As used herein, "comprises" and/or "comprising" refers to the presence of one or more other components, steps, operations and/or elements mentioned. or addition is not excluded.

Unless otherwise defined, all terms (including technical and scientific terms) used herein may be used with the meaning commonly understood by those of ordinary skill in the art to which the present invention belongs. In addition, terms defined in a commonly used dictionary are not to be interpreted ideally or excessively unless specifically defined explicitly.

Hereinafter, preferred embodiments of the present invention will be described in more detail with reference to the accompanying drawings. The same reference numerals are used for the same components in the drawings, and repeated descriptions of the same components are omitted.

The present invention relates to a cosmetic composition for improving acne prone skin comprising a cork oak extract. Specifically, it relates to a cosmetic composition for improving acne prone skin, comprising an extract of Quercus Suber bark and an extract of Quercus Root. And, the cosmetic composition may be prepared by mixing coconut (Cocos Nucifera) oil, thyme (Thymus Vulgaris) flower/leaf/stem extract, and camelina seed oil (Camelina sativa seed).

The cork oak (Quercus Suber) is cultivated in the Mediterranean region, and is an evergreen tree of the oak family, generally with a lifespan of 150-200 years. Cork is a spongy material that protects trees from fire, pests, and other external environments, and even if the bark of a cork oak tree is peeled off, it will be regenerated with better quality within 9 years. The cork oak is native to southern Europe and northern Africa, and usually survives in a Mediterranean climate and has a thick bark to prevent moisture evaporation. In the present invention, a cosmetic composition is prepared by mixing the bark (bark) of the cork oak.

Quercus does not refer to any one species, but to several species belonging to the genus Quercus. It means a useful tree with many uses, and all trees belonging to this genus produce nuts called acorns, so they are also called 'acorn trees'. There are deciduous broad-leaved trees that lose their leaves in winter and evergreen broad-leaved trees that do not lose their leaves all year round. 200-250 species grow from temperate to tropics in the northern hemisphere, and the leaves are alternate and most have serrated edges. Flowers are bisexual and bloom in April-May. The male flower heads hang down from the leaf axil of new branches, and the female flower heads stand upright at the top of the leaf axil. Nuts, called acorns, are housed in dish-like capsules and are oval or ball-shaped. Quercus aliena, Quercus variabilis, Quercus dentata, Quercus glauca, Quercus grosseserrata, Quercus mongolicodentata, and oak, as subspecies of the oak tree. Quercus acutissima, Quercus mongolica, Quercus serrata, Quercus myrsinaefolia, Quercus acuta, Quercus gilva, Cork oak, Quercus suber have. In the present invention, a cosmetic composition is prepared by mixing oak roots.

Thymus (Thymus Vulgaris) is a 100% wild organic plant belonging to the family Lamiaceae, the main components of which are thymol and carvacrol. These two specific polyphenols have strong antioxidant, antibacterial and anti-inflammatory properties. The complex of thyme and camelina virgin oil promotes the penetration of effective ingredients into the sebaceous glands. Camelina oil contains abundant linolenic acid (ω3) and linolenic acid (ω6), and acne-prone skin lacks linolenic acid. Thyme is distributed in Europe and Africa, and the origin is Southern Europe (Spain, Italy, France) and North Africa (Morocco). It has flowers and leaves, and in the present invention, a cosmetic composition is prepared by mixing the flower/leaf/stem extract of thyme.

Coconut palm is an evergreen tree belonging to the palm family. The fruit is coconut (Cocos Nucifera). The only broad palm tree in the genus of the coconut palm, it grows up to 30 m in length, with winged leaves 4 to 6 m long and small leaves 60 to 90 centimeters long. At the end of their lifespan, the fallen leaves are scattered around the tree, providing nutrients to the tree. Coconut means the fruit of the coconut palm. The leaves of the coconut palm are used as fuel or as a floor covering. Coconut fruit itself is used for food and is also used as oil and fat for palm oil and the like. In the present invention, a cosmetic composition is prepared by mixing coconut oil.

Camelina (Camellia sativa) is a flowering plant in the family Brassaceaceae, native to Europe and Central Asia. The plant is mainly grown as a seed crop in Europe and North America. The height of Camelina, an annual plant in summer and winter, is 30-120 cm, and the leaves and stems may be partially hairy. In England, it blooms from June to July. The abundant, four-petaled flowers are pale yellow and cruciform. Today, camelinas are found in almost all of Europe, Asia and North America, as well as in South America, Australia and New Zealand. Chamelena was traditionally grown as a seed crop to produce vegetable oil and animal feed. Camelina oil was used in oil lamps and edible oils. In the present invention, a cosmetic composition is prepared by mixing camelina seed oil.

The process of preparing a plant extract such as a cork oak extract, an oak root extract, and a thyme extract used in the cosmetic composition according to the present invention is as follows.

a) Let the plant to be extracted contain moisture and freeze with nitrogen in an environment below -80℃.

b) Grind frozen plants to form powder.

c) Mix the frozen powder with vegetable oil so that the effective ingredient of the plant to be extracted is included in the oil. At this time, since the plant contains a small amount of moisture before freezing, both the water-soluble and oil-soluble components of the plant are included.

d) In the process of making the effective component of the plant to be included in the vegetable oil, ultrasonic and high-frequency components are oscillated toward the extraction liquid to promote the extraction of the effective component of the plant.

e) When the extraction process is completed, it goes through a filtering process to finally produce an oil-based liquid plant extract.

The cosmetic composition of the present invention is used to improve acne-prone skin, and problematic skin, especially, acne-prone skin having symptoms such as excessive sebum production and oxidation of sebum, proliferation of Propionibacterium (cutibacterium) and P. acnes bacteria, hyperkeratosis and inflammation used to improve According to the cosmetic composition of the present invention, there are effects of preventing excessive production of sebum, limiting the proliferation of P. acnes, reducing inflammation, supplying moisture and increasing skin radiance.

The cosmetic composition of the present invention uses a plant extract and has strong anti-inflammatory, antioxidant and antibacterial effects. Cosmetically, the skin is less oily and non-greasy, and pores are improved and reduced. In addition, skin blemishes, acne and redness can be reduced, resulting in a smoother complexion.

The cosmetic composition of the present invention controls sebum production and inhibits the activity of 5-alpha reductase enzyme to prevent excessive sebum secretion. And, it reduces inflammation by limiting the proliferation of P. acnes, reducing IL-8 secretion by P. acnes, and inhibiting the production of lipid mediators of oxidized squalene-induced inflammation.

In addition, in the oil component of the cosmetic composition of the present invention, unsaturated fatty acids preserve the lipid balance of the skin and prevent keratinization disorders, thereby improving skin elasticity and healing. The oil component is a dry oil with high penetrability that makes the skin soft like silk. As a result, the skin is regenerated and balanced, making it elastic, soft and smooth.

Camelina oil of the cosmetic composition of the present invention is rich in linoleic acid and linolenic acid, which are essential for the formation of the epidermal lipid barrier. These are fatty acids that form cell membranes, and these polyunsaturated fatty acids act to inhibit inflammation. Linoleic acid also reduces blackheads and contributes to reduced skin irritation.

The cosmetic composition of the present invention treats acne by inhibiting inflammatory cytokines, inhibiting the activity of 5-alpha reductase (5-a-reductase) enzymes, and inhibiting P. acnes bacteria.

Cytokines are factors that regulate interactions between cells related to immunity, inflammation, or hematopoietic response, and include interleukin-1, interleukin-6, interleukin-8, tumor necrosis factor (TNF-a), and the like. Excessive or unregulated cytokine production and secretion mediates or exacerbates various diseases by disrupting immunomodulatory functions, such as allergic diseases, inflammatory diseases, and autoimmune diseases.

Clinical trial results according to Example 1

According to Example 1 of the present invention, a cosmetic composition was prepared by mixing 2% by weight of cork oak bark extract, 1% by weight of oak root extract, 0.5% by weight of coconut oil, 0.3% by weight of thyme extract, and 0.2% by weight of camelina seed oil. did

The cosmetic composition 110 according to Example 1 of the present invention was applied to acne skin twice a day for 3 weeks for women and men in their 10s and 30s to determine the degree of improvement in the skin condition and analyze the improvement effect of acne skin did Referring to [Table 1] below, the efficacy was evaluated by dividing each item into good (5 points) and bad (1 points).

Target of use spreadability absorbency Moisture Stimulation oily feeling degree of incense 16 year old female 5 5 5 5 5 5 22 year old woman 5 5 4 5 5 5 24 year old female 5 3 3 4 3 3 26 year old female 4 4 4 3 3 3 19 year old male 4 3 3 4 3 4 21 year old male 3 5 3 5 5 2 21 year old male 4 5 5 5 4 One 34 year old male 5 5 5 5 5 One average 4.375 4.375 4 4.5 4.125 3

Referring to [Table 1], the cosmetic composition 110 according to Example 1 of the present invention is generally satisfactory according to the evaluation of clinical test subjects, and has good spreadability, so that it can be applied evenly over the entire face with a small amount, It has a feeling of being well absorbed by acne skin as a whole. However, some subjects evaluated that the scent was strong and felt a little repulsive. Referring to FIG. 1 , acne suppression and improvement effects of the cosmetic composition 110 according to Example 1 of the present invention can be observed for acne-prone skin.

Clinical trial results according to Example 2

According to Example 2 of the present invention, a cosmetic composition was prepared by mixing 1% by weight of cork oak bark extract, 1% by weight of oak root extract, 0.5% by weight of coconut oil, 0.3% by weight of thyme extract, and 0.2% by weight of camelina seed oil. did

The cosmetic composition 120 according to Example 2 of the present invention is applied to acne skin twice a day for 3 weeks for women and men in their 10s and 30s for 3 weeks to determine the degree of improvement in the skin condition and analyze the improvement effect of acne skin did Referring to [Table 2] below, the efficacy was evaluated by dividing each item into good (5 points) and bad (1 points).

Target of use spreadability absorbency Moisture Stimulation oily feeling degree of incense teenage woman 3 4 3 5 3 2 woman in her twenties 4 4 4 4 3 3 woman in her twenties 4 4 3 3 3 5 woman in her twenties 4 5 5 5 5 4 teenage male 4 5 4 4 4 3 30's male 4 4 3 3 3 3 30's male 4 4 2 3 3 3 average 3.86 4.28 2.71 3.86 3.43 3.28

Referring to [Table 2], the cosmetic composition 120 according to Example 2 of the present invention is generally lacking compared to Example 1 according to the evaluation of clinical test subjects, and the spreadability is not as good as that of Example 1, The fragrance was evaluated as improved compared to Example 1. Referring to FIG. 2 , acne suppression and improvement effects of the cosmetic composition 120 according to Example 2 of the present invention can be observed for acne-prone skin.

Example of inhibitory effect on inflammatory cytokine production

TNF-α is a cytokine (glycoprotein) secreted during inflammation. Locally elevated concentrations of TNF-α produce signs suggestive of inflammation, leading to vasodilation associated with the presence of erythema (redness of the skin) and edema. TNF-α plays an important role in leading to inflammatory skin diseases such as psoriasis and atopic dermatitis.

We use the ARPE-19 cell line (adult retinal pigment epithelium-19), and since this cell model is particularly sensitive to inflammatory events, the tests are performed using the ARPE-19 cell line. These cells are produced from human retinal pigment epithelium (RPE). All tests are performed in a sterile environment in a laminar flow hood. Cells were seeded in 96-well microplates and then placed in an incubator for 24 hours to fuse with the test product upon incubation.

The amount of fluorescence signal was measured using a cell fluorescence meter. These monochromator analyzers can select the exact excitation and emission wavelengths to use for a particular sensor, so background noise is not biased. Each tested condition is repeated for three series of supernatants to verify the reproducibility of the experiment. Data are presented as percentage of fluorescence relative to negative control expressed in culture medium. The Dunnett test was followed by ANOVA (Analysis Of VAriance). [Table 3] shows the results of measuring cytokine TNF-α secretion by ARPE-19 cells after 1%, 2% and 3% treatment of the cosmetic composition (Example 1, Example 2) of the present invention, respectively.

Inhibition rate of inflammatory cytokine production (%) = 100-(Amount produced in the presence of each test substance/Amount of P.acnes-treated control group × 100)

Test substance (treatment concentration) Cytokine production inhibition rate (%) 1% extract of Example 1 44% Extract of Example 1 2% 63% Extract of Example 1 3% 68% 1% extract of Example 2 38% Extract of Example 2 2% 54% Extract of Example 2 3% 59% P.acne (control) 0%

5 α-reductase enzyme activity inhibitory effect test example

5 α-reductase is the most important enzyme in the biosynthesis of male hormones and is the most important factor in the generation of puberty acne through excessive sebum production. 5 By measuring the inhibitory effect of the test substance on α-reductase, the indirect effect on the inhibition of sebum production and acne can be confirmed.

When testosterone is converted to DHT by 5 α-reductase reductase, the sebaceous glands are stimulated and the secretion of sebum becomes severe. Progesterone is a precursor of androgens and estrogen, and when the secretion of progesterone increases, the amount of testosterone also increases. When excessive sebum and keratinocytes secreted from the sebaceous glands continue to form, they clog the pores (hair follicles) to form comedones. When P. acnes proliferates in the area where the comedones occur, acne is created by an inflammatory reaction.

The cosmetic composition of Example was treated with the culture medium of HEK293 (Human Embryonic Kidney) cell line, which is a human-derived 5 α-reductase-secreting cell, and it was measured whether the secretion of 5 α-reductase was inhibited by the treatment of the cosmetic composition of Example. 5. Culture of the HEK293 cell line, which is an α-reductase-secreting cell, is performed in a sterile environment in a laminar flow hood. Cells were cultured in a 96-well microplate at a concentration of 106 cells/well (cells/well), cell adhesion was confirmed, and test substances were added to the cell culture medium by concentration and cultured for 24 hours. 5 α-androstane-3, 5 α-androstane-3, by the action of the substrate 4-androstene-3,17-dione 5 α-reductase according to the method of Baerbel P. et al. (Mol Cell Biochem 270: 201-208, 2005) by recovering the culture medium. The rate of change to 17-dione was measured.

Enzyme activity inhibition rate (%) = 100- (production amount in the presence of each test substance / substrate change amount in control group × 100)

Test substance (treatment concentration) Enzyme activity inhibition rate (%) 1% extract of Example 1 62% Extract of Example 1 2% 78% Extract of Example 1 3% 90% 1% extract of Example 2 48% Extract of Example 2 2% 64% Extract of Example 2 3% 77% control 0%

Example of antibacterial effect against P. acnes bacteria

The antibacterial effect on P. acnes was measured according to the Kirby-Bauer standard method. Acne 106 cultured in anaerobic conditions in brain heart infusion broth mixed with 7 milliliters (m1) of plate medium (containing 0.8% phytoagar), dispensed with petridish and hardened. Then, a disk (filter paper disc) with a diameter of 10 mm (mm) soaked with the extraction composition of Example was placed on the surface of the medium, and incubated in anaerobic conditions at 37° C. for 24 hours. The antibacterial effect was confirmed by measuring the diameter of the clear zone where the growth of acne bacteria was suppressed, and the results are shown in [Table 5] below.

Test substance (treatment concentration) Clear Zone(mm) 1% extract of Example 1 14.2 Extract of Example 1 2% 18.7 Extract of Example 1 3% 24.1 1% extract of Example 2 12.3 Extract of Example 2 2% 15.8 Extract of Example 2 3% 21.4 control 0

Example of killing effect on P.acnes bacteria

Using the cosmetic composition 110 according to Example 1 and the cosmetic composition 120 according to Example 2 of the present invention, the P.acne bacteria were cultured in an anaerobic medium at a concentration of 0.5%, 1.0%, and 2.0%. After the injection, each death degree (size of growth inhibition ring) was evaluated, and the results are shown in [Table 6] (unit: cm).

At this time, the P.acne bacteria were cultured on an anaerobic plate, and then inoculated into an anaerobic broth medium, and cultured was used.

0.5% 1.0% 2.0% Extract of Example 1 2.1 2.7 3.3 Extract of Example 2 1.4 1.9 2.5 control 0.1 0.4 0.6

Referring to [Table 6], it can be seen that the killing effect of P.acne bacteria is excellent in Examples 1 and 2 according to the present invention, and in particular, in Example 1, P compared to Example 2 It can be seen that the killing effect of .acne bacteria is better.

NO (Nitro oxide) production inhibitory effect test example

Since NO (Nitro oxide) is one of the reactive oxygen species and plays an important role in inducing inflammation, the amount of NO produced was confirmed by using the griess reagent as follows.

First, RAW264.7 cells were adjusted to 1.0×105 cell/ml using DMEM medium, and then inoculated in a 48-well plate, and pre-cultured for 18 hours. The test substance and a fresh medium containing LPS (1 μg/ml) were simultaneously treated and cultured for 24 hours. 100 μl of the cell culture supernatant and 100 μl of Griess reagent were mixed and reacted for 10 minutes in a 96-well plate, and absorbance was measured at 540 nm using an ELISA reader. The amount of NO produced was measured in the form of NO ₂ ⁻ present in the cell culture medium using a reagent.

Test substance (treatment concentration) NO production (%) 1% extract of Example 1 57.2 Extract of Example 1 2% 53.8 Extract of Example 1 3% 48.3 1% extract of Example 2 74.6 Extract of Example 2 2% 69.2 Extract of Example 2 3% 63.1 control 89.7

Referring to [Table 7], both the cosmetic composition 110 according to Example 1 and the cosmetic composition 120 according to Example 2 of the present invention exhibited NO production inhibitory effects, Example 1 and Example Comparing 2, a better effect was exhibited in the case of Example 1. When the cosmetic composition 110 according to Example 1 is used, the NO production inhibitory activity is improved.

Experimental example of measuring antioxidant effect using NBT method

In order to confirm the antioxidant level of the cosmetic composition 110 according to Example 1 and the cosmetic composition 120 according to Example 2 of the present invention, BHT, which is well known as an antioxidant, was used as a comparative sample, and the antioxidant measurement was performed using the NBT method. did

The NBT method is a method of generating active oxygen by xanthine and xanthine oxidase, and measuring the blue color produced by reacting the produced active oxygen with nitro blue tetrazolium (NBT) at a wavelength of 560 nm, The active oxygen scavenging rate was measured, and the measurement method is as follows.

To a Bayer bottle, add 2.4 ml of 0.05M sodium carbonate (Na ₂ CO ₃ ), 0.1 ml of 3 mM xanthine solution, 0.1 ml of 3 mM EDTA solution, 0.1 ml of BSA solution, and 0.1 ml of 0.72 mM NBT solution, and add the sample solution to it. After each addition, it was left to stand at 25 degreeC for 10 minutes. After leaving for 10 minutes, 0.1 ml of xanthine oxidase solution was added, stirred rapidly, and then incubated at 25° C. for 20 minutes, and 0.1 ml of 6 mM copper chloride (CuCl ₂ ) solution was added to stop the reaction and absorbance at 560 nm. (St) was measured. In the blank test, distilled water was used instead of the sample solution, and absorbance (Bt) was measured in the same manner as above. For the blank of the sample solution, the absorbance (Bo) was measured in the same manner as above using distilled water instead of the xanthine oxidase solution.

The measurement result was calculated by the following formula.

Antioxidant effect (%) = [1-(St-So)/(Bt-Bo)] × 100

St: absorbance at 560 nm after the enzymatic reaction of the sample solution

So: Absorbance at 560 nm before the enzymatic reaction of the enzyme-free sample solution

Bt: Absorbance at 560 nm after enzymatic reaction of blank test solution

Bo: Absorbance at 560 nm before the enzymatic reaction of the enzyme-free blank test solution

Test substance (treatment concentration) Antioxidative effect(%) 1% extract of Example 1 67 Extract of Example 1 2% 78 Extract of Example 1 3% 91 1% extract of Example 2 55 Extract of Example 2 2% 69 Extract of Example 2 3% 83 Control (BHT 3%) 71

Referring to [Table 8], in the case of the cosmetic composition 110 according to Example 1 of the present invention, it was confirmed to have a better antioxidant effect at the same concentration as BHT, and from the above results, the active ingredient of the present invention The excellent antioxidant effect of the cosmetic composition including the cork oak bark extract, oak root extract, thyme extract, coconut oil and camelina seed oil was confirmed.

Experimental example for measuring the effect of scavenging active oxygen in cells

EGCG was used as a comparative sample to measure the scavenging effect of active oxygen generated in cells by ultraviolet rays of the cosmetic composition 110 according to Example 1 and the cosmetic composition 120 according to Example 2 of the present invention, The following experiment was performed using a fluorescent material.

Human keratinocytes HaCaT cell line (Human keratinocytes HaCaT cell line) was dispensed in 96-well plates for fluorescence measurement at 2 × 104 cells per well, and penicillin / streptomycin (penicillin / streptomycin) was added using DMEM medium at 37 ° C. After culturing for 1 day under 5% CO ₂ conditions, the extracts of Example 1 or Example 2 were treated at a concentration of 0.02, 0.01, or 0.005% by weight.

After adding the test substance and incubating for 24 hours, the medium was washed with HCSS (HEPES-buffered control salt solution) to remove the remaining medium, and 100 μl of DCFH-DA prepared at 20 μM was added to HCSS, followed by 20 μL at 37° C., 5% CO ₂ condition. After incubation for minutes, it was washed with HCSS. Thereafter, 100 μl of HCSS treated for each concentration was added, and the fluorescence of DCF (dichlorofluorescein) initially oxidized with active oxygen was measured with a fluorescence plate reader (Ex; 485 nm, Em; 530 nm). After UVB (20mJ/cm2) was irradiated and the extract of Example 1 or Example 2 was treated, the fluorescence was measured using a fluorescence plate reader (Ex; 485 nm, Em; 530 nm).

Test substance (treatment concentration) Active oxygen scavenging effect (%) 1% extract of Example 1 34 Extract of Example 1 2% 45 Extract of Example 1 3% 56 1% extract of Example 2 25 Extract of Example 2 2% 36 Extract of Example 2 3% 44 Control (EGCG 3%) 41

Referring to [Table 9], it was found that the cosmetic composition 110 according to Example 1 of the present invention had a higher active oxygen scavenging effect than EGCG at the same concentration as EGCG. From the above results, it was confirmed that the cosmetic composition containing the cork oak bark extract, oak root extract, thyme extract, coconut oil and camelina seed oil, which are the active ingredients of the present invention, has an excellent active oxygen scavenging effect.

Experimental example of measuring the cytotoxicity alleviation effect by UV irradiation

The alleviating effect of cytotoxicity by UV irradiation of the cosmetic composition 110 according to Example 1 and the cosmetic composition 120 according to Example 2 of the present invention was measured.

Fibroblasts (fibroblasts) were placed in a 24-well plate 1×105 each and allowed to adhere for 24 hours. Each well was washed once with PBS, and 500 μl of PBS was added to each well. After irradiating the fibroblasts with UV light of 10 mJ/cm 2 using an ultraviolet B (UVB) lamp, PBS was removed and 1 ml of cell culture medium (DMEM with 10% FBS added) was added. Here, the extract of Example 1 or Example 2 of the present invention was treated and then cultured for 24 hours. After 24 hours, the medium was removed, and 500 μl of cell culture medium and 60 μl of MTT solution (2.5 mg/ml) were added to each well, and then cultured in a 37° C. CO ₂ incubator for 2 hours. After incubation, the medium was removed and 500 μl of isopropanol-HCl (0.04N) was added. Then, the cells were lysed by shaking for 5 minutes, and 100 μl of the supernatant was transferred to a 96-well plate, and absorbance was measured at 565 nm using a microplate reader.

The cell viability (%) was measured by the following equation, and the cytotoxicity mitigation rate by ultraviolet light was calculated.

Cell viability (%) = [(St-Bo)/(Bt-Bo)] × 100

St: Absorbance at 565 nm of wells treated with test substances for color reaction

Bo: Absorbance at 565 nm of wells subjected to color reaction with cell culture medium only

Bt: Absorbance at 565 nm of wells that were not treated with test substances

Reduction of cytotoxicity by ultraviolet light (%) = [1-(St-Bo)/(Bt-Bo)] × 100

St: Cell viability of wells treated with UV irradiation and test substances

Bo: Cell viability of wells that were not irradiated with UV light and were not treated with test substances

Bt: Cell viability of wells irradiated with UV light and not treated with test substances

Test substance (treatment concentration) Cytotoxicity mitigation rate (%) 1% extract of Example 1 23 Extract of Example 1 2% 32 Extract of Example 1 3% 41 1% extract of Example 2 15 Extract of Example 2 2% 24 Extract of Example 2 3% 33 control 0

Referring to [Table 10], it was found that in the case of the cosmetic composition 110 according to Example 1 of the present invention, cytotoxicity caused by ultraviolet rays was effectively protected. From the above results, it was confirmed that the cosmetic composition containing the cork oak bark extract, oak root extract, thyme extract, coconut oil and camelina seed oil, which are the active ingredients of the present invention, has excellent cytotoxicity mitigation effect by UV irradiation, From that, it was found that the skin irritation alleviation effect of the present invention by ultraviolet rays was found.

Whitening effect experiment of cosmetic composition containing cork oak extract (Example 1)

Acne scars (acne scars) are likely to develop in acne sufferers. Acne Scars usually appear as skin bumps (pitted or uneven skin) or pigmentation. In general, even if inflammatory acne is cured, melanin is generated while the inflammation continues, and thus pigmentation may occur. The cosmetic composition comprising the cork oak extract according to Example 1 of the present invention has a whitening effect that helps to improve acne skin.

For the test method, prepare 0.1M sodium phosphate Buffer (pH 6.5), 0.1M sodium phosphate Buffer (pH 7.0), L-Tyrosine, and Tyrosinase solution, dilute the sample 50% with Ethanol, and perform sonication for 1 hour. After serial dilution with ethanol to 10%, 5%, 2%, and 1%, 880 μl of 0.1M sodium phosphate buffer (pH 6.5) is added to 80 μl of each sample. After centrifugation, filtration with a Syringe filter is used as a sample solution. The analysis equipment uses a VERSAmax microplate reader, and the detector uses 475nm (UV).

As a result of conducting a whitening effect test (tyrosinase activity inhibition ability) by changing a total of 5 concentrations in this experiment, it is confirmed that the activity inhibition rate decreases as the concentration decreases, as in the case of Arbutin used as a standard (refer to FIGS. 3 and 4).

After the tyrosinase activity inhibition reaction, a color change was observed as shown in FIG. 6 , and in the case of Arbutin, the higher the concentration, the closer to colorless, and the cosmetic composition of Example 1 was not colorless like Arbutin, but the color was most It is confirmed that it appears faintly.

The cosmetic composition of the present invention is used in a cosmetic composition for improving acne skin containing a cork oak extract as an active ingredient. In an embodiment according to the present invention, the cork oak bark extract is contained in the cosmetic composition in an amount of 2 to 2.5% by weight. The cosmetic composition of the present invention may be used in preparing a softening lotion, a nourishing lotion, a nourishing cream, a massage cream, an essence, an eye cream, a cleansing cream, a cleansing foam, a cleansing water, a pack, a spray or a powder.

In conclusion, the cosmetic compositions containing the cosmetic compositions according to Examples 1 and 2, respectively, show that there is an improvement effect on acne skin, but in particular, the cosmetic composition containing 2 wt % or more of the cork oak extract is effective for acne skin. It shows that the improvement effect is very excellent.

It is to be understood that the above-described embodiments are illustrative in all respects and not restrictive, and the scope of the present invention will be indicated by the following claims rather than the foregoing detailed description. And it should be construed that all changes and modifications derived from the meaning and scope of the claims as well as equivalent concepts are included in the scope of the present invention.

110: Cosmetic composition according to Example 1
120: the cosmetic composition according to Example 2

Claims (1)

A cosmetic composition for improving acne prone skin, comprising a cork oak bark extract, an oak root extract, a thyme extract, coconut oil and camelina seed oil.

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