KR20230080174A - Composition for anti-oxidation, anti-inflammation, and acne comprising extracts unripe fruit derived from Poncirus trifoliata - Google Patents

Abstract

The present invention relates to a composition containing immature tangerine fruit extract for anti-oxidation, anti-inflammation, and prevention and treatment of acne. An active ingredient of the present invention is an immature tangerine fruit extract, which is excellently safe to the human body without skin irritation or cytotoxicity, has high total polyphenol and flavonoid content, is excellent in DPPH radical scavenging activity, ABTs radical cation scavenging activity and SOD-like activity, inhibits cellular NO production, has good antimicrobial effects against various skin flora including P. acnes, improves skin barrier function, ameliorates and inhibits skin sebum secretion, and improves skin pores, so as to be useful as an active ingredient in cosmetic compositions, pharmaceutical compositions, topical skin care compositions, and quasi-drug compositions for anti-oxidation, anti-inflammation, and prevention, amelioration and treatment of acne.

Description

Composition for anti-oxidation, anti-inflammation, and acne comprising extracts unripe fruit derived from Poncirus trifoliata}

The present invention relates to a composition for preventing, improving, and treating anti-oxidation, anti-inflammatory, and acne, comprising an extract of immature Tangerine fruit.

Due to the recent COVID-19 crisis, masks have become essential consumer goods, and skin is easily sensitive due to the high temperature and high humidity environment in the mask and continuous friction, and skin troubles are occurring due to aggravation of inflammatory factors. In addition, if the non-woven fabric, preservative, adhesive, etc. used in the mask come into contact with sensitive skin, symptoms such as redness or peeling of the skin on the face, ears, etc. may occur due to contact dermatitis. The disease may also get worse. Referring to this phenomenon, a new word called 'maskne', which is a combination of mask and acne, has recently appeared.

The main cause of skin diseases such as skin damage is caused by the inflammatory reaction in the body, and nitric oxide (NO), a kind of reactive radical species in the body inflammatory reaction, is produced by immune cells such as macrophages. It is known to play an important role in various physiological and pathological processes. Nitric oxide is a very unstable compound and is produced through the activation of inducible nitric oxide synthase (iNOS), an enzyme that produces nitric oxide (NO), by an inflammatory substance such as lipopolysaccharide (LPS). COX (cyclooxygenase) is also called PGHS (prostaglandin H synthase), and COX-1 and COX-2 are currently known as important regulatory enzymes that act to convert membrane phospholipids into prostalandins (thromboxanes). . COX-1 is basically present in tissues and shows a certain action steadily, whereas COX-2 is expressed at the inflammatory reaction site of animals or humans, and is known to be rapidly expressed within a short period of time by various stimuli. It is known that an increase in the expression of COX-2, a representative inflammatory enzyme, also affects the production of nitric oxide (NO).

In order to improve or prevent such skin damage and aging, non-steroidal antioxidants such as benzidamine, intomethacin, ibuprofen, etc., or steroids such as dexamethasone and hydrocortisone, which prevent cell aging by inhibiting the generation of active oxygen, have conventionally been used. Antioxidants of the system were used to improve skin wrinkles and skin inflammatory diseases. In addition, a substance having inhibitory activity against derosinase, such as hydroquinone, ascorbic acid, kojic acid, or glutathione, was used to improve skin damage and aging. However, since the above chemicals cause side effects when used excessively and have problems with stability and safety, it is necessary to develop a composition derived from natural products that is safe and has excellent skin disease improvement effects with few side effects.

In addition, acne is an inflammatory disease of hair follicles and sebaceous glands, and is one of the most common skin diseases that continuously occur from teenagers to middle-aged people. Recently, adult acne has increased, and it has been reported that 40% of men and 54% of women (25 years of age or older), and 3% of men and 12% of women in middle-aged people have acne. Although the exact cause of acne is not known, it is generally caused by complex factors such as excessive sebum secretion from sebaceous glands, hormonal imbalance, genetic predisposition, abnormal keratinization in hair follicles, proliferation of Propionibacterium acnes , and inflammatory response.

Regarding acne, the United States and Japan show a lot of interest in the treatment or cosmetics sector, but they have released acne skin improvement products mainly based on chemical compounds. In this case, we are concentrating our efforts on developing cosmetics using natural materials.

Recently, the number of acne patients visiting dermatology hospitals and clinics is increasing, and considering the market prospect of acne medicines and treatment devices, it is predicted that the demand for acne cosmetics that can continuously care for patients other than medicines will increase due to the win-win effect. .

On the other hand, Jisil (枳實, Ponciri fructus ) is the immature fruit of the tangerine tree belonging to the Rutaceae family that is split in half and dried. It is used as a treatment for indigestion and gastrointestinal disorders by acting on the stomach and intestines to make stagnant Qi, phlegm, and food pass well. It has pharmacological effects that lighten the body, so it is used for the treatment of edema, constipation, postpartum abdominal pain, prolapse, and phlegm.

However, almost no modern research has been conducted on ginseng. Korean Patent Registration No. 10-1654737 discloses a composition for relieving hangover containing extracts of cinnamon and cinnamon as active ingredients, and Korean Patent No. 10-0920899 No. only discloses a cosmetic composition for converging pores containing a branch extract and HPC (Hydrogenated phosphatidylcholine) as active ingredients.

Accordingly, the inventors of the present inventors paid attention to the usefulness of the fruit of the tangerine tree and made diligent efforts to develop it as a natural cosmetic raw material. I will find out very good and complete the present invention.

One object of the present invention is to provide a cosmetic composition for preventing and improving anti-oxidation, anti-inflammatory, or acne, comprising an extract of immature Tangja fruit as an active ingredient.

Another object of the present invention is to provide a pharmaceutical composition for preventing and treating anti-inflammatory or acne, comprising an extract of immature Tangja fruit as an active ingredient.

Another object of the present invention is to provide a composition for external application for skin containing an extract of immature Tangja fruit as an active ingredient for antioxidant, anti-inflammatory, or prevention and improvement of acne.

Another object of the present invention is to provide a quasi-drug composition for preventing and improving anti-oxidation, anti-inflammatory, or acne, comprising an extract of immature Tangja fruit as an active ingredient.

In one aspect, the present invention provides a cosmetic composition for antioxidation, anti-inflammatory, or prevention and improvement of acne, comprising an extract of immature tangerine fruit as an active ingredient.

In the present invention, the immature Tangerine fruit refers to the unripe fruit of the Tangerine tree, an evergreen shrub plant of the Rutaceae family. In oriental medicine, the dried immature tangerine fruit is called a branch, and the mature tangerine fruit is called a crust, but in the present invention, the branch is included.

In the present invention, the immature Tangerine fruit extract refers to a product obtained by extracting dried, raw, cooked, or frozen immature Tangerine fruit, preferably obtained by extracting dried immature Tangerine fruit.

The immature Tangja fruit extract is (a) water, (b) anhydrous or hydrous lower alcohol having 1-4 carbon atoms (methanol, ethanol, propanol and butanol, etc.), (c) acetone, (d) ethyl acetate, (e) chloroform , (f) butyl acetate and (g) 1,3-butylene glycol, and was obtained from Jisil by using a solvent selected from the group consisting of 1,3-butylene glycol as an extraction solvent. The extraction solvent is preferably a solvent selected from the group consisting of water, hydrous lower alcohol, and 1,3-butylene glycol, more preferably water, hydrous 1,3-butylene glycol, or water, lower alcohol, and 1,3-butylene glycol. The hydrous lower alcohol may be 30-90% alcohol, preferably 40% alcohol, and the hydrous 1,3-butylene glycol may be 20-95% 1,3-butylene glycol alcohol, preferably 90% 1,3-butylene glycol or 20% 1,3-butylene glycol.

The suitable amount of the extraction solvent can be appropriately adjusted because the degree of extraction and loss of the active ingredient of the extract may differ, but it is 1-30 times the dry weight of the immature tangerine fruit, preferably 1-20 times, and more Preferably it is 5-20 times.

As one specific example, the immature Tangerine fruit extract of the present invention is dried by immature Tangerine fruit, washed with clean water, dried and finely powdered, and water as an extraction solvent 20 times the dry weight of the powdered immature Tangerine fruit, 90% 1,3-butylene glycol (1,3-butylene glycol 90: water 10 (v/v)), a mixture of 20% 1,3-butylene glycol and 30% ethanol (1,3-butylene Extraction was performed using 20 glycol: 40 water: 40 ethanol (v/v).

On the other hand, it is obvious to those skilled in the art that the extract of the present invention can be obtained by using not only the above extraction solvent, but also an extract of immature Tangja fruit exhibiting substantially the same effect using other extraction solvents.

In addition, the extract of the present invention includes not only the extract by the above-described extraction solvent, but also an extract subjected to a conventional purification process. For example, obtained through various additional purification methods, such as separation using an ultrafiltration membrane having a certain molecular weight cut-off value, separation by various chromatography (made for separation according to size, charge, hydrophobicity or affinity), etc. The fraction is also included in the immature Tangja fruit extract of the present invention.

The immature Tangja fruit extract of the present invention can be appropriately adjusted because the degree of extraction and loss of the active ingredient of the extract by the extraction solvent may differ, but preferably, cold extraction, ultrasonic extraction, reflux cooling extraction, pressurized extraction, An extraction method such as pressurized hot water extraction can be used.

As an example, when water is used as an extraction solvent for the immature tangerine fruit extract, a pressurized hot water extraction method may be used.

The active ingredient of the composition of the present invention, immature Tangja fruit extract, may be included in an amount of 0.001 to 50% by weight, more preferably 0.01 to 30% by weight, and most preferably 0.1 to 10% by weight based on the total weight of the composition. there is. At this time, if the content of the immature tangerine fruit extract is less than 0.001% by weight, the purpose of the present invention, such as antioxidant, anti-inflammatory and prevention, improvement, and treatment effects of skin acne, cannot be obtained, and if it exceeds 50% by weight, the increase in the content Depending on the composition, the effect is not proportional and may be inefficient, and there is a problem in that the stability of the formulation is not secured.

The active ingredient of the composition of the present invention, immature Tangja fruit extract, has high total polyphenol and flavonoid content, high DPPH radical scavenging activity, ABTs radical cation scavenging activity, and high SOD-like activity, so it has excellent antioxidant effect and is safe without cytotoxicity. It is material.

In addition, the extract of immature Tangerine fruit, which is an active ingredient of the composition of the present invention, effectively improves NO production of LPS-induced macrophages, thereby improving, preventing, and treating inflammation.

Here, "prevention" means any action that inhibits or delays the onset of oxidative stress, inflammation, or acne of the skin by administration of the cosmetic composition according to the present invention, and "improvement" means any action by administration of the cosmetic composition It refers to all activities that improve or beneficially change the symptoms of oxidative stress, inflammation, or suspected or affected individuals of acne.

In addition, the immature tangerine fruit extract, which is an active ingredient of the composition of the present invention, has an excellent antibacterial effect against various skin flora including acne bacillus, improves skin barrier function, improves and suppresses skin sebum secretion, and reduces skin pores. It is effective in preventing, improving, and treating acne on the skin.

Therefore, the immature Tangja fruit extract of the present invention can be used as an active ingredient of a cosmetic composition for antioxidation, anti-inflammatory, and prevention and improvement of acne skin, which can be antioxidant, anti-inflammatory, and prevent and improve acne skin.

In the present invention, the ingredients included in the "cosmetic composition" may include ingredients commonly used in cosmetic compositions in addition to the active ingredients as active ingredients, such as antioxidants, stabilizers, solubilizers, vitamins, pigments and It may include conventional adjuvants such as flavoring agents, and carriers. The cosmetic composition of the present invention can be prepared in any formulation conventionally prepared in the art, for example, a solution, suspension, emulsion, paste, gel, cream, lotion, powder, soap, surfactant-containing cleansing , It may be formulated into oil, powder foundation, emulsion foundation, wax foundation, pack, massage cream and spray, etc., but is not limited thereto. More specifically, it may be prepared in the form of softening lotion, nutrient lotion, nourishing cream, massage cream, essence, eye cream, cleansing cream, cleansing foam, cleansing water, pack, spray or powder.

When the formulation of the present invention is a paste, cream or gel, animal oil, vegetable oil, wax, paraffin, starch, tracanth, cellulose derivative, polyethylene glycol, silicone, bentonite, silica, talc or zinc oxide may be used as a carrier component. can

When the formulation of the present invention is a solution or emulsion, a solvent, solubilizing agent or emulsifying agent is used as a carrier component, such as water, ethanol, isopropanol, ethyl carbonate, ethyl acetate, benzyl alcohol, benzyl benzoate, propylene glycol, 1 ,3-butyl glycol oil, fatty acid esters of glycerol, polyethylene glycol or sorbitan.

When the formulation of the present invention is a suspension, as a carrier component, a liquid diluent such as water, ethanol or propylene glycol, an ethoxylated isostearyl alcohol, a suspending agent such as polyoxyethylene sorbitol ester and polyoxyethylene sorbitan ester, microcrystals Star cellulose, aluminum metahydroxide, bentonite, agar or tracanth and the like may be used.

When the formulation of the present invention is a powder or spray, lactose, talc, silica, aluminum hydroxide, calcium silicate or polyamide powder may be used as a carrier component, and in particular, in the case of a spray, additional chlorofluorohydrocarbon, propane / May contain a propellant such as butane or dimethyl ether.

When the formulation of the present invention is surfactant-containing cleansing, as carrier components, aliphatic alcohol sulfate, aliphatic alcohol ether sulfate, sulfosuccinic acid monoester, isethionate, imidazolinium derivative, methyl taurate, sarcosinate, fatty acid amide ether Sulfate, alkylamidobetaine, aliphatic alcohol, fatty acid glyceride, fatty acid diethanolamide, vegetable oil, lanolin derivative, or ethoxylated glycerol fatty acid ester may be used.

When the cosmetic composition of the present invention is a soap, a surfactant-containing cleansing formulation, or a surfactant-free cleansing formulation, it may be applied to the skin and then wiped off, removed, or washed with water. As specific examples, the soap is liquid soap, powder soap, solid soap, and oil soap, the surfactant-containing cleansing formulation is a cleansing foam, cleansing water, cleansing towel, and cleansing pack, and the surfactant-free cleansing formulation is a cleansing cream. , cleansing lotion, cleansing water and cleansing gel, but are not limited thereto.

In another aspect, the present invention provides a pharmaceutical composition for preventing and treating inflammation or acne, comprising an extract of immature Tangja fruit as an active ingredient.

In the pharmaceutical composition of the present invention, the anti-inflammatory and preventive and therapeutic effects of the immature Tangja fruit extract, which is an active ingredient, and acne are as described above.

Here, "prevention" refers to any action that inhibits or delays the onset of inflammation or acne by administration of the pharmaceutical composition according to the present invention, and "treatment" refers to suspected inflammation or acne by administration of the pharmaceutical composition. and all actions that improve or beneficially change the symptoms of an affected subject.

In the present invention, the prevention or treatment of inflammation and acne is accomplished by suppressing cellular NO production, suppressing the activity of acne bacillus, improving skin barrier, and improving skin sebum and pores with the immature tangerine fruit extract. It can be.

The pharmaceutical composition containing the immature Tangerine fruit extract, which is an active ingredient of the composition of the present invention, is 0.001 to 70% by weight, more preferably 0.01 to 70% by weight, most preferably 0.1 to 10% by weight based on the total weight of the composition. It can be included in weight percent. At this time, if the content of the immature tangerine fruit extract is less than 0.001% by weight, anti-inflammatory and skin acne prevention and treatment effects cannot be obtained, and if it exceeds 70% by weight, the effect is not proportional to the increase in the content and is inefficient It may be, and there is a problem that the stability of the formulation is not secured.

In the present invention, "pharmaceutical composition" means prepared for the purpose of preventing or treating a disease, and may be formulated and used in various forms according to conventional methods. For example, it can be formulated into oral formulations such as powders, granules, tablets, capsules, suspensions, emulsions, syrups, and aerosols, and diluents or excipients such as lubricants, wetting agents, flavoring agents, emulsifying agents, suspending agents, preservatives, and surfactants It can be formulated and used as a parenteral formulation using. In addition, it may be formulated and used in the form of external preparations, suppositories, external preparations for the skin, and sterile injection solutions.

In addition, according to each formulation, pharmaceutically acceptable carriers such as buffers, analgesics, solubilizers, isotonic agents, stabilizers, and bases may be prepared by further including carriers known in the art.

In the present invention, "pharmaceutically acceptable carrier" may mean a carrier or diluent that does not inhibit the biological activity and properties of the active ingredient to be injected while not stimulating living organisms. The type of the carrier that can be used in the present invention is not particularly limited, and any carrier commonly used in the art and pharmaceutically acceptable can be used. Non-limiting examples of the carrier include saline, sterile water, Ringer's solution, buffered saline, albumin injection solution, dextrose solution, maltodextrin solution, glycerol, ethanol, and the like. These may be used alone or in combination of two or more. The carrier may be a non-naturally occurring carrier.

The composition of the present invention can be administered in a pharmaceutically effective amount. The pharmacologically effective amount means an amount that is sufficient to treat a disease at a reasonable benefit/risk ratio applicable to medical treatment and does not cause side effects. It may be determined according to the activity of the drug, sensitivity to the drug, method of administration, time of administration, route of administration and excretion rate, duration of treatment, factors including drugs used in combination or concurrently, and other factors well known in the medical field. Specifically, it is generally 0.001 mg to 300 mg per day per 1 kg of the body weight of the administered subject, and may be administered once or several times a day at regular time intervals according to the judgment of a doctor or pharmacist, but is not limited thereto.

In addition, the pharmaceutical composition of the present invention may be used alone or in combination with other pharmaceutically active compounds exhibiting effects of preventing, improving, or treating inflammation or acne, or in a suitable set.

The pharmaceutical composition of the present invention may be administered as an individual therapeutic agent or in combination with other therapeutic agents, and may be administered sequentially or simultaneously with conventional therapeutic agents. And it can be single or multiple administrations. It is important to administer the amount that can obtain the maximum effect with the minimum amount without causing side effects in consideration of all of the above factors, and can be easily determined by those skilled in the art.

As used herein, the term "administration" means introducing the pharmaceutical composition of the present invention to a subject by any suitable method, and the administration route of the composition of the present invention is various oral or parenteral routes as long as it can reach the target tissue. can be administered through

The administration method of the pharmaceutical composition according to the present invention is not particularly limited, and may follow a method commonly used in the art. As a non-limiting example of the administration method, the composition may be administered by oral administration or parenteral administration. Preferably parenteral administration, more preferably it can be applied by topical application (topical application) by application.

As another aspect, the present invention provides a method for preventing or treating inflammation or acne, comprising administering a pharmaceutical composition containing an extract of immature Tangja fruit.

The preventive and therapeutic effects of the immature Tangja fruit extract on inflammation and acne are as described above.

As used herein, the term "subject" may refer to all animals, including humans, who have or are likely to develop inflammatory or acne skin diseases. The animal may be not only humans but also mammals such as cattle, horses, sheep, pigs, goats, camels, antelopes, dogs, and cats that require treatment for similar symptoms, but are not limited thereto.

Specifically, the preventive or therapeutic method of the present invention may include administering the composition in a pharmaceutically effective amount to a subject having or at risk of developing inflammatory or acne skin disease. The administration method is as described above.

The prevention may mean any action that suppresses or delays the onset of inflammatory and acne skin diseases by administering the composition for preventing, improving, or treating inflammation and acne according to the present invention to a subject.

In addition, the treatment may refer to any action that improves or benefits the symptoms of inflammation or acne by administering the composition of the present invention to a subject suspected of having inflammation or acne.

The pharmaceutical composition containing the immature tangerine fruit extract is as described above.

For administration, the composition of the present invention may contain pharmaceutically acceptable carriers, excipients or diluents in addition to the above-described active ingredients. The carriers, excipients and diluents include lactose, dextrose, sucrose, sorbitol, mannitol, starch, acacia gum, alginate, gelatin, calcium silicate, microcrystalline cellulose, polyvinylpyrrolidone, cellulose, water, syrup, methyl cellulose, methylhydroxybenzoate, propylhydroxybenzoate, talc, magnesium stearate and mineral oil.

An appropriate dosage of the composition of the present invention varies depending on the condition and weight of the patient, the severity of the disease, the type of drug, and the time, but can be appropriately selected by those skilled in the art. Specifically, the dose of the immature Tangja fruit extract may be 0.001 to 300 mg/Kg. In addition, in the case of an external preparation, it is recommended to apply it 1 to 5 times a day in an amount of 1.0 to 3.0 ml on an adult basis and continue for at least one month. However, the dosage is not intended to limit the scope of the present invention.

In another aspect, the present invention provides a composition for external application for skin containing an extract of immature Tangja fruit as an active ingredient for antioxidant, anti-inflammatory, or prevention and improvement of acne.

Antioxidant, anti-inflammatory, or acne preventing or improving effects of the immature Tangja fruit extract are as described above.

The term "improvement" of the present invention refers to skin diseases caused by oxidative stress, inflammation, or acne caused by an increase in the expression and related activity of oxidative stress, inflammation, or acne in a subject, skin diseases caused by inflammation, or suppression of skin diseases caused by acne, It means alleviation, elimination.

The composition for external application for skin according to the present invention may be formulated by containing a cosmetically or dermatologically acceptable medium or base. These are all formulations suitable for topical application, for example solutions, gels, solids, anhydrous pasty products, emulsions obtained by dispersing an oil phase in an aqueous phase, suspensions, microemulsions, microcapsules, microgranules or ionic forms (liposomes) and It may be provided in the form of a non-ionic vesicular dispersion or in the form of a cream, toner, lotion, powder, ointment, spray, pack or conceal stick. It can also be used in the form of a foam or in the form of an aerosol composition further containing a compressed propellant. These compositions can be prepared according to conventional methods in the art.

In addition, the composition for external application for skin according to the present invention includes a fatty substance, an organic solvent, a solubilizing agent, a thickening agent, a gelling agent, a softening agent, an antioxidant, a suspending agent, a stabilizer, a foaming agent, a fragrance, a surfactant, water, and an ionic type. or nonionic emulsifiers, fillers, sequestering agents, chelating agents, preservatives, vitamins, blocking agents, wetting agents, essential oils, dyes, pigments, hydrophilic or lipophilic actives, lipid vesicles or any other commonly used in cosmetics. It may contain adjuvants commonly used in the field of cosmetology or dermatology, such as ingredients. The adjuvant is introduced in an amount generally used in the field of cosmetology or dermatology.

In another aspect, the present invention provides a quasi-drug composition for antioxidation, anti-inflammatory, or prevention or improvement of acne, comprising an extract of immature Tangja fruit as an active ingredient.

Antioxidant effect, anti-inflammatory effect, and prevention and treatment effect of acne of the immature Tangja fruit extract are as described above.

The term “quasi-drugs” of the present invention refers to articles with milder effects than pharmaceuticals among articles used for the purpose of diagnosing, treating, improving, mitigating, treating or preventing human or animal diseases. For example, quasi-drugs under the Pharmaceutical Affairs Act exclude items used for pharmaceutical purposes, and include products used for the treatment or prevention of human/animal diseases, products with minor or no direct action on the human body, and the like.

Specifically, the quasi-drug may include external skin preparations and personal hygiene products. More specifically, it may be disinfectant cleaner, shower foam, gagreen, wet tissue, detergent soap, hand wash, or ointment, but is not limited thereto.

When the composition according to the present invention is used as a quasi-drug additive, the composition may be added as it is or used together with other quasi-drugs or quasi-drug ingredients, and may be appropriately used according to a conventional method. The mixing amount of the active ingredient may be appropriately determined depending on the purpose of use.

Since the immature tangerine fruit extract of the present invention is made from natural medicinal plants as raw materials, it can be used as a cosmetic composition, pharmaceutical composition, external skin composition, and quasi-drug composition for preventing, improving, or treating skin diseases such as antioxidant, anti-inflammatory, and/or acne. Even when used, it may have fewer side effects compared to general synthetic compounds, so it can be safely included and used usefully.

The composition containing the immature tangerine fruit extract of the present invention as an active ingredient is excellent in human body stability due to no skin irritation and cytotoxicity, and is very effective in antioxidant, anti-inflammatory, and prevention, improvement, and treatment of acne. It can be usefully used as an active ingredient in cosmetic compositions, pharmaceutical compositions, compositions for external application for skin, and quasi-drug compositions for preventing, improving, and treating acne.

1 is a water extract (PPW), water and 1,3-butylene glycol mixed solvent extract (PBG), water, ethanol and 1,3-butylene glycol mixed solvent of immature Tangja fruit according to an embodiment of the present invention. It is a figure showing the total polyphenol and total flavonoid contents for the extract (PME).
2 is a water extract (PPW), water and 1,3-butylene glycol mixed solvent extract (PBG), water, ethanol and 1,3-butylene glycol mixed solvent of immature Tangja fruit according to an embodiment of the present invention. This is a picture showing the DPPH radical scavenging ability of the extract (PME).
Figure 3 is a water extract (PPW), water and 1,3-butylene glycol mixed solvent extract (PBG), water, ethanol and 1,3-butylene glycol mixed solvent of immature Tangja fruit according to an embodiment of the present invention This is a picture showing the ABTs radical scavenging ability for the extract (PME).
4 is a water extract (PPW), water and 1,3-butylene glycol mixed solvent extract (PBG), water, ethanol and 1,3-butylene glycol mixed solvent of immature Tangja fruit according to an embodiment of the present invention. It is a figure showing SOD-like activity for extract (PME).
5 is a diagram showing the results of measuring the viability of Raw 264.7 macrophages and skin keratinocytes, HaCaT cells, induced by LPS for water extract (PPW) of immature Tangja fruit according to an embodiment of the present invention.
6 is a measurement of the viability of Raw 264.7 macrophages and skin keratinocytes, HaCaT cells, in response to water and 1,3-butylene glycol mixed solvent extract (PBG) of immature tangerine fruit according to an embodiment of the present invention. This is the picture showing the result.
Figure 7 shows the survival rate of Raw 264.7 macrophages and skin keratinocytes, HaCaT cells, in response to water, ethanol, and 1,3-butylene glycol mixed solvent extract (PME) of immature tangerine fruit according to an embodiment of the present invention. This figure shows the measured results.
8 is a water extract (PPW), water and 1,3-butylene glycol mixed solvent extract (PBG), water, ethanol and 1,3-butylene glycol mixed solvent of immature Tangja fruit according to an embodiment of the present invention. It is a figure showing the degree of inhibition of NO production of macrophages induced by LPS for extract (PME).
9 is a water extract (PPW), water and 1,3-butylene glycol mixed solvent extract (PBG), water, ethanol and 1,3-butylene glycol mixed solvent of immature Tangja fruit according to an embodiment of the present invention. It is a figure showing the degree of TNF-α secretion inhibitory activity of macrophages induced by LPS for extract (PME).
10 is water extract (PPW), water and 1,3-butylene glycol mixed solvent extract (PBG), water, ethanol and 1,3-butylene glycol mixed solvent of immature Tangja fruit according to an embodiment of the present invention. It is a figure showing the degree of IL-1β secretion inhibitory activity of macrophages induced by LPS for extract (PME).
11 is a water extract (PPW), water and 1,3-butylene glycol mixed solvent extract (PBG), water, ethanol and 1,3-butylene glycol mixed solvent of immature Tangja fruit according to an embodiment of the present invention. This is a picture showing the antibacterial effect of the extract (PME).
12 is a picture taken using Antera 3D as a result of a clinical test of a cosmetic composition containing a water extract of immature Tangja fruit according to an embodiment of the present invention.
13 is a picture showing the results of a survey on efficacy after 4 weeks of use of the test product as a result of a clinical test of a cosmetic composition containing a water extract of immature Tangja fruit according to an embodiment of the present invention.
14 is a picture showing the results of a survey on the quality after 4 weeks of use of the test product as a result of a clinical test of a cosmetic composition containing a water extract of immature Tangja fruit according to an embodiment of the present invention.

Hereinafter, examples and the like will be described in detail to aid understanding of the present invention. However, the embodiments according to the present invention can be modified in many different forms, and the scope of the present invention should not be construed as being limited to the following examples. Embodiments of the present invention are provided to more completely explain the present invention to those skilled in the art.

Example 1: Preparation of immature tangerine fruit extract

The immature fruit extract of the tangerine tree used dried immature fruit of the tangerine tree, which was purchased from "Okcheondang", a company specializing in herbal medicines. Distilled water corresponding to 20 times (w/w) of the weight of the cell was added and extracted with a pressure extractor (DM-701, Korea) at 110 ° C and 1.5 atm for 3 hours to obtain a pressure heating water extract (PPW). , Water and 1,3-butylene glycol diluted in a ratio of 9:1 were used to extract the eggplant at 60° C., which was used as a PBG extract (1,3-butylene glycol in water extract). In addition, water, ethanol, and 1,3-butylene glycol were mixed at a volume ratio of 4:4:2, respectively, and extracted at 60° C. as a solvent to obtain water and ethanol, 1,3-butylene glycol extract (PME). Each extract of the three samples extracted with different solvents was filtered with filter paper (Whatman No 2), and the filtered extract was concentrated under reduced pressure by 1/10 with a rotary vacuum evaporator (Eyela 400 series, Tokyo, Japan). After that, it was used as a sample for measuring useful components and physiological activity.

Example 2: Determination of total polyphenol and total flavonoid content of immature tangerine fruit extract

The content of polyphenolic compounds in the extracts of immature Tangja fruit extracted under different conditions as in Example 1 was measured by the Folin-Denis (AOAC, 2005) method. To 0.2 mL of the sample, 1.8 mL of distilled water and 0.2 mL of Folin-Ciocalteu's phenol reagent (Junsei Chemical Co. Tokyo, Japan) were added, mixed, and reacted for 3 minutes. Then, 0.4 mL of a saturated Na ₂ CO ₃ solution was added and mixed. After adding 1.4 mL of distilled water and reacting at room temperature for 1 hour, the absorbance was measured at 725 nm using a spectrophotometer (Mecasys Optizen POP, Korea). The content of polyphenolic compounds contained in each extract was obtained from the tannic acid calibration curve.

The total content of flavonoids was determined by modifying the method of Nieva Moreno et al. (2000), adding 0.1 mL of 10% aluminum nitrate, 0.1 mL of 1 M potassium acetate, and 4.7 mL of 80% ethanol to 0.1 mL of the sample, and reacting at 25 ° C for 40 minutes. The absorbance was measured at 415 nm, and the total flavonoid compound content of the immature Tangja fruit extract extracted under different conditions was obtained from the quercetin calibration curve.

The results of measuring the contents of polyphenols and flavonoid compounds contained in three extracts extracted by mixing immature Tangja fruit with water, 1,3-BG and water, and water with ethanol and 1,3-BG are shown in Table 1 and Figure 1. Same as PBG extracted by mixing equal amounts of water and 1,3-BG contained the most polyphenolic compounds at 117.87 mg/g, and PME, a mixed extract of water, ethanol, and 1,3-BG, and PPW, a pressurized hot water extract, had 67.15 mg/g, respectively. It contained mg/g and 61.33 mg/g of polyphenolic compounds. However, the flavonoid compound contains PPW 80.74 mg/g > PME 45.31 mg/g > PBG 20.70 mg/g, and the pressurized hot water extract extracted at high temperature and high pressure has antiseptic and moisturizing effects, so it is used as a cosmetic solvent. Compared to the extracts containing BG, the content of flavonoid compounds was high.

Extract Compound contents (mg/g) Total polyphenols Total flavonoids PPW 61.33±2.96 ^c 80.74± ^5.83a PBG 117.87± ^2.24a 20.70± ^3.51c PME 67.15± ^0.56b 45.31± ^1.29b

^abc All values represent the mean ± standard deviation of triplicate measurements, and different letters in the superscript within the same column are significantly different at p < 0.05 by Duncan's multiple range test.

Example 3: DPPH radical scavenging activity

The hydrogen donation effect on DPPH (1,1-diphenyl-2-picryl hydrazyl) was measured according to Blois' method (1958). 1 mL of DPPH diluted in absolute ethanol at a concentration of 0.2 mM was added to 2 mL of immature tangerine fruit extract sample, mixed, and reacted at 37 ° C for 30 minutes. indicated by

The results are shown in Table 2 and FIG. 2 below. At a concentration of 2,000 ug/mL, PME was 105.84%, which showed better DPPH radical scavenging rate than natural antioxidant L-ascorbic acid, and PPW and PBG were also 96.51% and 96.39%, respectively, similar to L-ascorbic acid in DPPH radical scavenging effect. showed All extracts showed a scavenging effect of 50% or more at a concentration of 125 ug/mL, although lower than that of the control L-ascorbic acid, and PME showed a scavenging rate of 65%. And PPW showed a similar or higher scavenging effect compared to the control L-ascorbic acid even at concentrations of 500 ug/mL and 1,000 ug/mL.

Extracts Concentration (ug/mL) 125 250 500 1,000 2,000 PPW 57.76± ^0.96c 73.89± ^3.28b 88.45± ^1.88a 92.42±1.30 ^a 96.51± ^2.05b PBG 53.80± ^1.43d 64.07±3.73 ^d 66.20±0.70 ^c 82.69± ^2.33d 96.39± ^0.28b PME 65.51± ^1.00b 68.72±1.49 ^c 73.74± ^1.61b 89.88± ^1.37b 105.84± ^1.61a AsA 82.22±2.44 ^a 85.52±1.33 ^a 87.54± ^0.32a 87.97±0.49 ^c 96.91± ^0.49b

AsA: L-ascorbic acid

^abc All values represent the mean ± standard deviation of triplicate measurements, and different letters in the superscript within the same column are significantly different at p < 0.05 by Duncan's multiple range test.

Example 4: ABTs radical cation scavenging activity

ABTs ⁺ free radicals are removed by antioxidants in the extract, and the antioxidant activity is measured by using the decolorization of blue-green, a unique color of radicals, using the ABTs cation decolorization assay method (Re et al., 1999). did Specifically, 7 mM 2,2-azino-bis (3-ethyl benzthiazoline-6-sulfonic acid) and 2.4 mM potassium persulfate were mixed and allowed to stand for 24 hours in the dark at room temperature to form ABTs ⁺ , and then diluted with ethanol. After adding 100 uL of sample to ABTs ⁺ 100 uL and allowing it to stand for 1 minute, the absorbance was measured at 732 nm, and the difference between the absorbance of the treated and untreated groups was expressed as a percentage (%) as ABTs radical cation scavenging ability.

The results for this are shown in Table 3 and Figure 3. At a concentration of 2,000 ug/mL, PPW, a pressurized hot water extract of immature fruits of Tangerine japonica, showed similar activity to L-ascorbic acid, a control group, and PBG, a 1,3-BG extract, had 50.31%, water and ethanol, 1,3 -BG mixed extract, PME, showed 77.56% ABTS radical scavenging rate. Even at a concentration of 500 ug/mL, PPW showed more than 50% activity, showing the best activity among the three extracts.

Extracts Concentration (ug/mL) 125 250 500 1,000 2,000 PPW 29.42±2.92 ^c 40.94± ^1.46b 64.84± ^1.49b 84.24± ^0.70b 97.27± ^1.43b PBG 37.61± ^3.71b 41.82± ^3.03b 42.00±1.88 ^c 49.77±3.40 ^c 50.31±2.78 ^d PME 20.56± ^2.11d 25.09±0.77 ^c 35.32±1.74 ^d 46.26±2.68 ^d 77.56±2.75 ^c AsA 99.33±0.75 ^a 99.01±0.47 ^a 99.42± ^0.43a 100.45± ^0.41a 100.94±0.40 ^a

AsA: L-ascorbic acid

^abc All values represent the mean ± standard deviation of triplicate measurements, and different letters in the superscript within the same column are significantly different at p < 0.05 by Duncan's multiple range test.

Example 5: SOD (Superoxide dismutase)-like activity

It is commercially important as it can suppress various diseases or aging involving superoxide by acting as a defense against active oxygen damage in vivo. The SOD-like activity of three extracts of immature fruit of the tangerine tree was measured by using the autoxidation reaction to pyrogallol for the reaction to substances similar to SOD, which can be expected to have anti-aging effects.

Specifically, the SOD-like activity was measured by adding 2.6 mL of tris-HCl buffer corrected to pH 8.5 and 0.2 mL of 7.2 mM pyrogallol to 0.2 mL of the sample according to the method of Marklund and Marklund (1975), reacting at 25 ° C for 10 minutes, and then reacting at 1 The reaction was stopped by adding 0.1 mL of N HCl. The amount of oxidized pyrogallol in the reaction solution was measured for absorbance at 420 nm, and the difference in absorbance between the added and non-added groups was expressed as a percentage (%).

The results are shown in Table 4 and Figure 4. Immature Tangja fruit extract, PPW 16.03% > PME 6.39% > PBG 5.48% at a concentration of 2,000 ug/mL, the pressurized hot water extract extracted at high temperature and high pressure has about 2.5 to 2.9 times higher SOD-like activity than water, BG extract, and the mixed extract. , and there was no reaction at concentrations of 500 ug/mL or less.

Extracts Concentration (ug/mL) 500 1,000 2,000 PPW 6.07±2.22 ^b2) 12.13± ^1.11b 16.03± ^0.96b PBG 3.28± ^1.05c 5.41±2.71 ^c 5.48±1.99 ^c PME - 4.93±1.45 ^c 6.39±1.24 ^c AsA 99.12± ^0.15a 99.74± ^0.26a 100.26±0.46 ^a

AsA: L-ascorbic acid

^abc All values represent the mean ± standard deviation of triplicate measurements, and different letters in the superscript within the same column are significantly different at p < 0.05 by Duncan's multiple range test.

Example 6: Cytotoxicity evaluation

MTT (3-[4,5-dimethylthiazol-2-yl]-2,5-diphenyltetrazolium bromide) assay was performed on RAW264.7 cells, a mouse macrophage cell line, using the immature fruit extract of Tangerine sinensis. Survival rate was expressed as percentage (% control). RAW264.7 macrophages were cultured in Dulbecco's modified Eagle's medium (DMEM) containing 10% fetal bovine serum (FBS) and 1% antibiotic-antimycotic (10 units/mL penicillin, 100 μg/mL streptomycin, 0.25 μg/mL) at 37 °C. It was cultured in an incubator (Forma series Ⅱ, Water Jacketed M311, Thermo Electron Co., USA) supplied with 5% CO ₂ at ℃.

200 μL of RAW 264.7 cells were dispensed into a 96-well plate (SPL, Pocheon, Korea) at a concentration of 1×10 ⁵ cells/mL, and after 24 hours, 100 μL of DMEM medium containing the diluted extract was added to the cells. After incubation for 24 hours, 100 μL of 5 μg/mL MTT (Sigma-Aldrich Co., St Louis, MO, USA) solution was added to each well, and after incubation for 3 hours, the supernatant was removed and each well After dissolving formazan by adding 100 μL of DMSO (dimethyl sulfoxide), the absorbance was measured at 570 nm using ELISA (Xmark microplate absorbance spectrophotometer, Bio-Rad, XMarkTM, Japan), and the cell viability for the control was expressed as a percentage (% control). ).

For HaCaT, a human keratinocyte cell line, the MTT assay was performed on the extract of immature fruit of Tangerine japonica in the same manner as above, and the cell viability for the control group was expressed as a percentage (% control).

The results for this are shown in Figures 5 to 7. In Raw 264.7 macrophages, at a concentration of 200 ug/mL, all extracts showed a survival rate of over 85%, and in HaCaT cells, at a high concentration of 200 ug/mL, PPW, a pressurized hot water extract, showed a survival rate of over 90%. More than 85% of the cells survived in PBG extracted by mixing PBG with 1,3-BG, water and ethanol, and PME extracted with 1,3-BG as a solvent. As a result of the above results, the three extracts of Jisil, immature fruits of Tangja tree with different extraction solvents, did not show toxicity affecting Raw 264.7 macrophages and HaCaT keratinocytes.

Example 7: Nitric oxide (NO) generation inhibitory effect

In order to confirm the immune enhancing ability of the immature fruit extract of Tangja tree, the concentration of NO produced in the culture supernatant of RAW 264.7 cells was measured using a microplate assay. RAW 264.7 macrophages were divided into 1×10 ⁵ cells in a 6-well plate, cultured for 24 hours, and the extract was treated by concentration (20, 50, 100 ug/mL), and as a positive control, 1 ug/mL of lipopolysaccharide (LPS) and cultured for 24 hours. After mixing 100 uL of cultured cell supernatant with the same amount of Griess regent (Sigma, St. Louis, MO, USA) and reacting at room temperature for 15 minutes, ELISA reader (Xmark microplate Absorbance Spectrophotometer, Bio-Rad, Hercules, CA, USA) Absorbance was measured at 540 nm using The degree of inhibition of NO production was calculated by obtaining the difference between the absorbance value of the experimental group measured after treating the selected extract for each concentration and the absorbance value of the control group treated only with LPS.

The results for this are shown in FIG. 8 . The LPS-treated group showed slightly more NO production than the LPS-untreated group, and all extracts had lower NO production than the control group, and NO production was inhibited by about 16% or more at a concentration of 200 ug/mL. Even at a concentration of 25 ug/mL, NO production was reduced by about 10%.

Example 8: Measurement of inflammation-related cytokine production

Macrophages are cells that maintain homeostasis in the body by being involved in innate and acquired immunity. Their activity is increased by immune enhancers such as interferon (IFN) and lipopolysaccharide (LPS), and they are tumor necrosis factor-increasing immune responses. α (TNF-α), interleukin (IL)-1β, IL-6, and nitric oxide (NO) are secreted (Hibbs et al., 1998).

The amount of cytokines (TNF-α, IL-1β) secreted from the culture supernatant of Raw 264.7 macrophages treated with immature tangerine fruit extract with LPS (1 ug/mL) was measured by ELISA cytokine kit (R&D system, Minneapolis, MN, USA). ) was measured using.

The results are shown in Figures 9 and 10. As can be seen here, at a concentration of 200 μg/mL, the hot-water extract (PPW), 1,3-BG extract (PBG), and the mixed extract (PME) had about 10% of TNF-α secretion inhibitory activity. shown (FIG. 9). In addition, in the IL-1β secretion inhibition experiment, PPW showed inhibitory activity of about 28%, PME 25%, and PBG 17%, so that the hot water extract of immature Tangerine sinensis showed the highest inhibition rate in inhibition of interleukin secretion (FIG. 10). Since the immature Tangja tree extract has an effect on suppressing inflammation in the body, it is considered to have an effect on suppressing acne.

Example 9: Antibacterial effect

Three extracts of immature Tangerine fruit were obtained from the skin flora Staphylococcus aureus (KCCM 41331), Staphylococcus epidermidis (KCCM 35494), and Escherichia coli (KCCM 40271) in liquid medium and solid medium, nutrient broth (NB) and nutrient agar (NA). was used, and was used as a strain for this experiment while culturing at 37 ° C. Propionibacterium acnes 3314, an acne bacterium, was activated by anaerobic culture in a 37°C incubator for 3 days using a reinforced clostridial medium (RCM), and then used as a strain for this experiment.

The antibacterial activity was measured by taking 1 platinum of each strain cultured in the medium by the clear zone and paper disc method, culturing the bacteria to be used for the experiment in 10 mL of liquid medium for 18 to 24 hours, and then activating them. , Inoculate 0.1 mL of bacterial solution into 10 mL of liquid medium, incubate for 3 to 6 hours, and then inoculate so that the number of bacteria per plate medium is about 1.7 × 10 ⁶ ~ 7.2 × 10 ⁷ cfu / mL, and smear evenly with a sterile cotton swab. . A sterilized filter paper disc (Tokyo, 6 mm, Japan) is placed on a solid plate medium, and the sample is absorbed by concentration to 0.05 mL/disc, and cultured for 18-24 hours at 37 ℃ for skin flora and acnes bacteria. The surrounding clear zone (mm) was confirmed.

The results of this are shown in FIG. 11 . Specifically, in the skin flora S. aureus , each extract showed the best clear zone with PBG 9.9 mm at a concentration of 20%, PPM and PPW showed growth inhibition of 8.9 mm and 8.8 mm, and PPW Even at a concentration of 10%, an inhibitory ring of 6.5 mm was displayed. In S. epidermidis , PBG was 6.7 mm and PPW was 6.2 mm, and PME did not show any effect. In E. coli bacteria, PBG 11.9 mm > PME 11.4 mm > PPW 1.0 mm inhibition ring was observed, and in the acne-causing bacteria P. acnes , growth inhibition ring of PBG 7.3 mm and PPW 7.0 mm was confirmed, and PPW was 10% Even at the concentration, a growth inhibition ring of 6.5 mm was shown.

Example 10: Acne skin condition improvement effect verification

Cosmetics containing the high-pressure hot water extract (PPW) of immature Tangja fruit were prepared with the composition shown in Table 5 below and commissioned to the Human Skin Clinical Trial Center Co., Ltd. Suitable for acne skin use, skin barrier function improvement, and skin sebum improvement , Skin pore improvement efficacy was evaluated.

Specifically, 20 healthy Korean men and women between the ages of 14 and 40 with acne lesions visited the Human Skin Clinical Trial Center, washed their faces with lukewarm water, and then treated them with constant temperature and humidity (22ㅁ2ㅀC, 50ㅁ 5%) condition, waiting for more than 30 minutes to sufficiently stabilize the skin, and then participate in the test. Before using the test product and after 4 weeks of use, a dermatologist visually evaluated the suitability for acne skin use, and the amount of transepidermal water loss in the cheek area (g/m ² h), the amount of sebum in the forehead area (μg/cm ² ), and the cheek area The mean pore volume (mm 3 ) of was analyzed to evaluate skin barrier function improvement, skin sebum improvement, and skin pore improvement efficacy.

award raw material name weighing amount (% by weight)) A Purified water 70.310 EDTA-2Na 0.03 1,2-Hexanediol 2.00 allantoin 0.10 1,3-BG 2.00 MSM 3.00 B hyaluronic acid 0.05 C Purified water 10.00 Carbomer 941 0.20 D Purified water 2.00 arginine 0.20 E ethanol 10.00 Azulene 0.01 Sym Clariol 0.10 F eggplant extract 0.10

10-1. Results of macroscopic evaluation of inflammatory and non-inflammatory lesions

A dermatologist who did not participate in the clinical trial counted the number of inflammatory lesions, and as a result of counting inflammatory and non-inflammatory lesions, no lesions were observed after 4 weeks of use of the test product, and comedones, pustules, and nodules were significantly decreased (Table 6).

10-2. Acne severity grading results

As a result of the dermatologist's evaluation of acne severity according to the Investigator's Global Assessment (IGA) criteria, the acne severity rating score decreased significantly after 4 weeks of using the test product (Table 7).

10-3. Skin barrier function improvement efficacy evaluation result

Compared to before using the test product, the amount of transepidermal water loss (g/m ² h) in the cheek area significantly decreased after 4 weeks of use (Table 8).

10-4. Skin Sebum Improvement Efficacy Evaluation Result

Compared to before using the test product, the amount of sebum (ug/cm ² ) on the forehead was significantly reduced after 4 weeks of use (Table 9).

10-5. Skin pore improvement efficacy evaluation result

Compared to before using the test product, after 4 weeks of use, the average pore volume (mm ³ ) of the cheek area was significantly reduced (Table 10).

10-6. Survey evaluation result after 4 weeks of test product use

As a result of conducting and analyzing the questionnaire on efficacy and quality after using the test product for 4 weeks to the test subjects, the efficacy satisfaction rate was 70 to 82%, and the quality satisfaction rate was 65 to 90%, indicating that the efficacy and quality were excellent.

10-7. Adverse reaction evaluation result

As a result of evaluating the skin reactions observed and recorded by the dermatologist during the test period and the skin reactions reported by the test subjects, early termination of the test due to temporary or intermittent reactions during the test period, the level of change in the method of use and frequency of the test It was judged that it was not a skin reaction.

10-8. Compliance evaluation result

After using the test product for 4 weeks, the test product use diary written by the test subject was collected and the compliance was evaluated. During this test period, all test subjects showed 99.8% compliance with the test product.

10-9. final result

After using the test product for 4 weeks, the test subjects were evaluated for suitability for acne skin use, skin barrier function improvement, skin sebum improvement, and skin pore improvement efficacy. As a result, dermatologists who did not participate in the clinical trial counted the number of inflammatory lesions. After 4 weeks of use, no lesions were caused, and comedones, pustules, and nodules were significantly reduced. In addition, the severity score of acne significantly decreased, and the amount of transepidermal water loss in the cheek area (g/m ² h), sebum amount in the forehead area (ug/cm ² ), and average pore volume in the cheek area (mean pore volume, mm ³ ) was significantly decreased. In conclusion, it was confirmed that the immature Tangja fruit extract according to the present invention is suitable for use on acne-prone skin, improves skin barrier function, improves skin sebum, and is effective in improving skin pores.

Claims (5)

A cosmetic composition for antioxidation, anti-inflammatory, or prevention or improvement of acne skin, comprising an extract of immature Tangerine fruit. The method of claim 1, characterized in that the immature tangerine fruit extract is extracted using water, 90% 1,3-butylene glycol, or a mixture of 20% 1,3-butylene glycol and 40% ethanol as a solvent. cosmetic composition. A pharmaceutical composition for anti-inflammatory or preventing or treating acne skin, comprising an extract of immature Tangerine fruit. A composition for external application for antioxidant, anti-inflammatory, or preventing or improving acne skin, comprising an extract of immature Tangerine fruit. A quasi-drug composition for antioxidant, anti-inflammatory, or prevention or improvement of acne skin, comprising an extract of immature Tangja fruit.

Images