KR101870143B1 - Composition for skin whitening or anti-inflammatory comprising Zizyphus jujube seed extract as effective component - Google Patents

Abstract

The present invention relates to a skin whitening composition or an inflammatory composition comprising a jujube extract as an active ingredient, wherein the extract of the jujube extract of the present invention solubilizes insoluble fibrous components through enzymatic treatment and ultrahigh pressure homogenization, And the jujube seed, which is a by-product after processing the jujube, can be efficiently utilized, thereby securing the competitiveness in terms of the manufacturing cost. The composition for skin whitening using the jujube extract effectively inhibits the production of melanin , A functional cosmetic for skin whitening, and a health functional food or skin pigmentation disease which can prevent or improve skin pigmentation disease. In addition, since it effectively inhibits NO production, it can be usefully used as a therapeutic agent for a health functional food or an inflammatory disease that can prevent or improve an inflammatory disease.

Description

[0001] The present invention relates to a skin whitening or anti-inflammatory composition comprising a jujube extract, as an active ingredient, a composition for skin whitening or anti-inflammatory,

The present invention relates to a skin whitening or anti-inflammatory composition comprising a jujube extract as an active ingredient.

The skin may be exposed to various harmful substances such as air and ultraviolet ray caused by air pollution and may cause various skin troubles. Among them, dandruff, freckles, and dandruff are among the biggest skin problems, and many efforts have been made to improve them.

Several factors are involved in determining the skin color of a person. Among them, the activity of melanocyte which makes melanin pigment, distribution of blood vessels, thickness of skin, carotenoid, bilirubin and other factors related to pigment are important. In particular, the most important factor is melanin, which is produced by the action of various enzymes such as tyrosinase in melanocytes in the human body. Skin pigmentation is caused by stimuli such as ultraviolet rays or inflammation, and it is also known to be caused by factors such as genetic factors, hormones, foods, and chemicals. In particular, inflammation related factors such as prostaglandins, histamine, alpha-MSH, and NO (nitric oxide) have been implicated in skin pigmentation abnormalities such as spots, freckles and pigmentation, and melanocytes ) Is known to activate melanin production. It is also reported that hyperpigmentation is accompanied by immunological mediators in tissue damage and regeneration process. When melanocytes are stimulated by various factors, tyrosinase produces melanin in the melanosome. As a result, melanocytes are melanosomes. That is, tyrosine in melanocytes is transformed into dopa by the action of tyrosinase and then transformed into dopaquinone by the action of dopaoxidase, and then tyrosinase-related related protein-2 (hereinafter referred to as TRP-2) and tyrosinase-related protein-2 (hereinafter referred to as TRP-2) to produce melanin. In this series of reactions, tyrosinase activates the rate-limiting reaction of the melanin pigmentation process, so that melanogenesis is primarily controlled by the activity and expression of tyrosinase. TRP-2, also known as the DOPA chromium mutant enzyme, activates the reaction of DOPA chromium with 5,6-dihydroxyindole-2-carboxylic acid (DHICA), TRP-1 activates DHICA with carboxylindole- . As a result, TRP-1 and TRP-2 are involved in the polymerization reaction, which is the final stage of melanin formation, an insoluble pigment. It has been reported that tyrosinase, tyrosinase-related protein-1 (TRP-1) and tyrosinase-related protein-2 (TRP-2) are activated by inflammatory mediators to increase the synthesis of melanin .

Currently developed whitening agents include kojic acid, arbutin, glutathione and vitamin C, but consumers do not have a satisfactory whitening effect and their use is limited due to skin safety. Thus, various attempts have been made to synthesize melanin.

In addition, inflammation is a normal defense mechanism of a living body that occurs when a physical injury or microorganism is infected. This inflammation neutralizes or eliminates the pathogen, To allow normal structure and function. Most of the diseases with inflammation result in deterioration of quality of life such as tissue damage, pain and itching, and chronic inflammatory conditions include arthritis, asthma, multiple sclerosis of brain and spinal cord, inflammatory bowel disease and arteriosclerosis ≪ / RTI > Macrophages are the main cells responsible for innate immunity. They are activated by a number of factors such as cytokines and bacterial lipopolysaccharide (LPS), and activated macrophages are nitric oxide (NO) and prostaglandin E ₂ (prostaglandin E ₂ , PGE ₂ ) as well as proinflammatory cytokines such as TNF-α, IL-6 (interleukin-6) and IL-1 (interleukin-1). NO is a free radical that is synthesized by nitric oxide synthase (NOS) and is composed of three isoforms of endothelial NOS, nNOS, and iNOS (inducible NOS) isoform) (Cho et al., 2014. Biomol. Ther. 22: 288-294). Proinflammatory cytokines such as IL-1, TNF-a, and interferon-gamma can induce iNOS expression and cause NO production. Many malignant cancers, such as brain cancer, breast cancer, lung cancer, prostate cancer, pancreatic cancer and melanoma, are associated with iNOS expression, and excessive production of NO can lead to epithelial cell carcinoma, mutations, and damage to DNA structures (Guo et al. Agric. Food Chem. 60: 2157-2164). Generally COX protein COX-1 and COX-2 in two kinds present the same type proteins (isoforms), and is PGE ₂ is produced by COX-2 during the inflammatory response (such as Jeon, 2014. Arch Pharm Res 37:. .. 907 -15). Expression of COX-2 is also highly correlated with tumorigenesis, such as cell damage, pain, edema, and heat, neovascularization, and metastasis. NF-κB is a transcription factor that plays an important role in the inflammatory response. In response to LPS, macrophages activate Myd88 (myeloid differentiation factor 88) by TLR4 (Toll-like receptor 4). Activated Myd88 activates transforming growth factor-activated protein kinase 1 (TAK1) and TAK1 activates NF-κB through degradation of IκB (inhibitor of NF-κB). NF-κB ultimately leads to inflammation by secretion of inflammatory mediators (Kawai et al., 2006. TLR signaling, Cell Death Differ. 13, 81625).

On the other hand, studies for improving the efficacy and extraction yield of the materials reported so far have shown improvements in yield due to increase in surface area due to fine grinding in the physical aspect, application of ultrasonic waves and homogenization for cell disruption, increase in solubility And biologically, fermentation and enzyme treatment processes for the solubilization of insoluble materials have been reported, but the ability of the material to improve the extraction efficiency and yield has been relatively limited. Recently, the improvement effect of extraction yield through hydrostatic hydrostatic pressure treatment is impossible to apply to mass production by batch type process, and it shows limit to improvement of extraction yield.

Ultra high pressure homogenization technology is a technique of passing a dispersion solution (hydrocolloid) through a micro-orifice module under ultra-high pressure (100 MPa or more) pressure, resulting in cavitation and high shear It is a technique to convert a dispersion solution into a homogeneous phase by deagglomeration and disconnection (depolymerization, low molecular weight) of the dispersions in the dispersion solution with a sudden increase in physical energy. Particularly, it is known that the fiber is made into a homogeneous dispersed phase and is effective for low molecular weight of protein and non-aggregated carbohydrate polymer.

Industrial applications using enzymes have been applied in various industrial fields, and in particular, in the field of food industry, it has become common to use pectinolytic enzymes to inhibit the formation of precipitates in fruit juice processing. However, many attempts have been made to improve the extraction yield and low molecular weight of insoluble components such as cellulose in the extraction process for the materialization, but it is not generalized. In particular, the fiber of the plant has a complex structure of fibrous structure, which is not easy to hydrolyse through a single enzyme treatment, because many fibrous tissues often form a complex with lignin in the fibrous tissue. However, with the help of various nano crushing technology, ultrahigh pressure homogenization technique and nano plasma treatment technology recently introduced, research for solubilizing insoluble polymer materials including cellulose by low molecular weight has been continuously carried out.

On the other hand, jujube has been used for many years to treat various diseases, such as the beauty effect that enriches the skin as well as the body's health by eating the date of jujube in the past, the effect of stabilizing the blood and stabilizing the mind, deciphering various toxins and antiallergic action It is known that jujube contains a large amount of vitamin C and trace elements, so it is said that it has effects such as smooth and clean skin, healthy skin and beautiful face. Especially, in jujube seeds which are used after processing jujube into processed foods, betulin and betulic acid are known as medicinal ingredients. In particular, c-AMP ingredient is about 1000 times higher than that of common plants, and other sugars and organic acids Nutritional components are known, and antioxidant, anticancer, blood glucose control, and anti-inflammatory effects have been reported.

As an example of a technique related to cosmetics, health functional foods or pharmaceutical compositions containing jujube seeds as an active ingredient, there is a soap using a jujube extract disclosed in Korean Patent No. 10-1070382 and a method for producing the same, 0801902 discloses an antimutagenic and cancer cell growth inhibiting composition containing a jujube extract. However, no technique for skin whitening or antiinflammatory composition containing the jujube extract of the present invention as an active ingredient has been disclosed.

The present invention provides a skin whitening or antiinflammatory composition comprising a jujube extract as an active ingredient. The extract of the jujube, which is an effective ingredient, The present invention has been accomplished by confirming the inhibition of the production of melanin and the anti-inflammatory effect as well as the extraction yield.

In order to accomplish the above object, the present invention provides a skin whitening cosmetic composition comprising an extract of a jujube produced by a method comprising the step of adding an enzyme to a mixture of water and a juice of a jujube seed, Compositions or anti-inflammatory compositions.

The present invention also provides a pharmaceutical composition for preventing or treating a melanin pigment hyperpigmentation disease comprising the above-mentioned jujuba extract as an active ingredient.

The present invention also provides a health functional food composition for preventing or ameliorating hyperpigmentation of melanin pigment comprising a jujube extract as an active ingredient.

The present invention also provides a pharmaceutical composition for preventing or treating an inflammatory disease comprising an extract of Daechu as an active ingredient.

The present invention also provides a health functional food composition for preventing or ameliorating an inflammatory disease comprising an extract of Daechu as an active ingredient.

The jujube seed extract of the present invention is obtained by solubilizing insoluble fibrous components through enzyme treatment and ultrahigh pressure homogenization treatment to improve the extraction yield and physiological activity and to efficiently utilize the by-products that are discarded, Since the composition for skin whitening using the jujube extract effectively inhibits the production of melanin, functional cosmetics for skin whitening and a therapeutic agent for a health functional food or skin pigmentation disease that can prevent or improve skin pigmentation disease It can be useful.

In addition, the anti-inflammatory composition using the jujube extract effectively inhibits the production of nitric oxide (NO), and thus can be usefully used as a therapeutic agent for a health functional food or an inflammatory disease that can prevent or improve an inflammatory disease.

FIG. 1 is a graph showing cytotoxicity, amount of melanin biosynthesis, and tyrosinase activity for examining skin whitening efficacy of the jujube extract of the present invention.
FIG. 2 is a graph showing cytotoxicity and nitric oxide (NO) production to examine the anti-inflammatory effect of the jujube extract of the present invention.

In order to achieve the object of the present invention,

A cosmetic composition for skin whitening comprising as an active ingredient a jujube extract prepared by a method comprising the steps of:

(a) preparing a jujuba seed powder by drying and pulverizing the jujube seeds;

(b) hydrolyzing the mixture of the jujube seed powder prepared in step (a) with water by adding an enzyme to the mixture;

(c) treating the hydrolyzate of step (b) with 100% ethanol; And

(d) homogenizing the ethanol-treated hydrolyzate of step (c) by ultra-high pressure.

In the method for producing the jujube extract of the present invention, the enzyme of step (b) may be Viscozyme or Pectinex. The viscozyme is a commercially available enzyme, Aspergillus a complex enzyme comprising arabinase, cellulase, Beta-glucanase, hemicellulase and xylanase derived from aculeatus , and the pectinex ) Is also a commercial enzyme and has an activity of pectinase and hemicellulase.

According to one embodiment of the present invention, the jujube seed powder can be obtained by increasing the yield of jujube seed extract and inhibiting melanin synthesis and NO production by high-pressure homogenization after treatment with viscose or pectinex.

In addition, in the method for producing the jujube seed extract of the present invention, the step (b) preferably comprises adding 10 to 14 times (v / w) of water adjusted to pH 3.5 to 5.5 to the jujube seed powder, (Viscozyme) or Pectinex and then hydrolyzed at 30 to 60 ° C for 1 to 12 hours. More preferably, the jujube seed powder is adjusted to pH 4.5 by 12 times (v / w) ) Was added to the mixture, and the mixture was hydrolyzed at 50 ° C for 8 to 12 hours or 12 times (v / w) of water adjusted to pH 4.5 in the juice of the jujubu seed. To the mixture was added Pectinex ) And then hydrolyzed at 37 ° C for 1 to 3 hours. The hydrolysis of the juice of the jujube seeds under the above conditions was able to improve the extraction yield by improving the extraction yield and improving the elution of the active material by insolubilizing the insoluble fraction of the jujube seeds or solubilizing them through molecular rearrangement.

In the method for producing the jujube extract of the present invention, the step (c) of ethanol treatment preferably comprises adding 6 to 8 times (v / v) of 100% ethanol to the hydrolyzate, To 3 hours. More preferably, the hydrolyzate can be treated by adding 100% ethanol 7 times (v / v) at 20 to 30 ° C for 2 hours with stirring at 180 rpm.

In addition, in the method for producing a jujube extract of the present invention, the ethanol-treated hydrolyzate may further include centrifuging and recovering the supernatant.

Also, in the method for preparing a jujube extract of the present invention, the ultra-high pressure homogenization of step (d) can be homogenized at a pressure of 100-120 MPa, and more preferably at a pressure of 120 MPa.

The method for producing the jujube extract of the present invention is more preferably

(a) preparing a jujuba seed powder by drying and pulverizing the jujube seeds;

(b) Viscozyme or Pectinex was added to a mixture prepared by adding 10 to 14 times (v / w) of water adjusted to pH 3.5 to 5.5 to the jujube seed powder prepared in step (a) Followed by hydrolysis at 30 to 60 ° C for 1 to 12 hours;

(c) adding 6 to 8 times (v / v) 100% ethanol to the hydrolyzate of step (b) and treating the mixture at 20 to 30 ° C for 1 to 3 hours; And

(d) homogenizing the ethanol-treated hydrolyzate obtained in step (c) under a pressure of 100 to 120 MPa.

In the present invention, it is possible to improve the extraction yield of the active material by making the fiber of the jujube seed extract into a homogeneous dispersed phase by enzyme treatment and ultrahigh pressure homogenization treatment and low molecular weight and solubilizing the insoluble dietary fiber.

On the other hand, the whitening effect refers to an effect to improve or prevent glaring skin, spots, freckles, and dark circles caused by various factors such as exposure to ultraviolet rays, change in hormone balance, genetic program, Or an effect of maintaining a beautiful skin with a sense of transparency and an effect of reducing the dullness of the skin to increase gloss and tautness. In general, it is known that the skin, spots, freckles, and dark circles of the skin, stamens, freckles and dark circles are stimulated by melanocytes due to stimulation by ultraviolet rays or changes in hormone balance, and melanin pigment is deposited on the skin. Therefore, if the production of melanin can be inhibited, it is possible to prevent or improve gross skin, spots, freckles and dark circles.

The cosmetic composition for skin whitening may be a solution, a suspension, an emulsion, a paste, a gel, a cream, a lotion, a powder, a soap, a surfactant-containing cleansing oil, a powder foundation, an emulsion foundation, a wax foundation and a spray , But the present invention is not limited thereto. The cosmetic composition comprising each of these formulations may contain various bases and additives necessary for formulation of the formulation, and the kinds and amounts of these ingredients can be easily selected by those skilled in the art.

The cosmetic composition of the present invention may contain, in addition to the active ingredient, at least one skin whitening active ingredient exhibiting the same or similar functions. Skin whitening active ingredients include kojic acid and its derivatives, arbutin, ascorbic acid and its derivatives, hydroquinone and its derivatives, resorcinol, cycloalkanone, methylenedioxyphenylalkanol, 2,7-dinitroindazole or Plant extracts such as mandarin orange extract, rice extract, licorice extract, and the like, but are not limited thereto.

When the formulation of the cosmetic composition of the present invention is a paste, a cream or a gel, an animal fiber, a plant fiber, a wax, a paraffin, a starch, a tragacanth, a cellulose derivative, a polyethylene glycol, a silicone, a bentonite, Etc. may be used.

When the formulation of the cosmetic composition of the present invention is a powder or a spray, lactose, talc, silica, aluminum hydroxide, calcium silicate or polyamide powder may be used as a carrier component. Especially, in the case of a spray, Propellants such as carbon, propane-butane or dimethyl ether.

When the formulation of the cosmetic composition of the present invention is a solution or an emulsion, a solvent, a solvent or an emulsifier is used as a carrier component, and examples thereof include water, ethanol, isopropanol, ethyl carbonate, ethyl acetate, benzyl alcohol, benzyl benzoate, Glycol, 1,3-butyl glycol oil, glycerol aliphatic ester, polyethylene glycol or fatty acid esters of sorbitan.

When the formulation of the cosmetic composition of the present invention is a suspension, a carrier such as water, a liquid diluent such as ethanol or propylene glycol, a suspension such as ethoxylated isostearyl alcohol, polyoxyethylene sorbitol ester and polyoxyethylene sorbitan ester, Microcrystalline cellulose, aluminum metahydroxide, bentonite, agar or tracant, etc. may be used.

When the formulation of the cosmetic composition of the present invention is a cleansing agent containing an interfacial active agent, the carrier component may include aliphatic alcohol sulfate, aliphatic alcohol ether sulfate, sulfosuccinic acid monoester, acethionate, imidazolinium derivative, methyltaurate, sarcosinate , Fatty acid amide ether sulfate, alkylamidobetaine, aliphatic alcohol, fatty acid glyceride, fatty acid diethanolamide, vegetable oil, linolenic derivative or ethoxylated glycerol fatty acid ester.

The cosmetic composition of the present invention may further contain an excipient including a fluorescent substance, a fungicide, a hygroscopic substance, a moisturizer, a fragrance, a fragrance carrier, a protein, a solubilizer, a sugar derivative, a sunscreen, a vitamin, .

The present invention also provides a pharmaceutical composition for preventing or treating melanin pigment hyperproliferative disorder or inflammatory disease, which contains, as an active ingredient, a jujube extract prepared by a method comprising the steps of:

(a) preparing a jujuba seed powder by drying and pulverizing the jujube seeds;

(b) hydrolyzing the mixture of the jujube seed powder prepared in step (a) with water by adding an enzyme to the mixture;

(c) treating the hydrolyzate of step (b) with 100% ethanol; And

(d) homogenizing the ethanol-treated hydrolyzate of step (c) by ultra-high pressure.

The method for preparing the extract of the jujube, which is an effective ingredient of the pharmaceutical composition of the present invention, is as described above. In addition to the jujube extract, the pharmaceutical composition preferably further comprises a carrier, an excipient or a diluent, but is not limited thereto.

As used herein, the term " melanin pigment hyperpigmentation "means darkening or darkening compared to other areas due to excessive increase of melanin in specific areas of skin or hands and toenails.

The melanin pigment hyperpigmentation disorder can be selected from the group consisting of freckles, senile spots, liver, spots, brown or black spots, daylight pigmentation, cyanic melasma, hyperpigmentation after drug use, gravidic chloasma, Post-inflammatory hyperpigmentation due to burns or dermatitis including burns, and the like.

The inflammatory disease is selected from the group consisting of dermatitis, allergy, atopic, asthma, conjunctivitis, periodontitis, rhinitis, otitis, sore throat, tonsillitis, pneumonia, gastric ulcer, gastritis, Crohn's disease, colitis, gout, ankylosing spondylitis, rheumatic fever, lupus, fibromyalgia, Wherein the disease is selected from the group consisting of psoriasis, psoriatic arthritis, osteoarthritis, rheumatoid arthritis, periarthritis, tendinitis, hay fever, tendinitis, myositis, hepatitis, cystitis, nephritis, sjogren's syndrome, multiple sclerosis and acute or chronic inflammatory diseases But is not limited thereto.

The pharmaceutical composition of the present invention may be various oral or parenteral formulations. In the case of formulation, a diluent or excipient such as a filler, an extender, a binder, a wetting agent, a disintegrant, or a surfactant is usually used. Solid formulations for oral administration include tablets, pills, powders, granules, capsules, and the like, which may contain one or more excipients such as starch, calcium carbonate, sucrose or lactose lactose, gelatin and the like. In addition to simple excipients, lubricants such as magnesium stearate, talc, and the like may also be used. Liquid preparations for oral administration include suspensions, solutions, emulsions, syrups and the like. Various excipients such as wetting agents, sweeteners, fragrances, preservatives and the like may be included in addition to water and liquid paraffin, which are simple diluents commonly used. have. Formulations for parenteral administration include sterile aqueous solutions, non-aqueous solvents, suspensions, emulsions, freeze-dried preparations, and suppositories. Examples of non-aqueous solvents and suspensions include propylene glycol, polyethylene glycol, vegetable oils such as olive oil, injectable esters such as ethyl oleate, and the like. Examples of suppositories include witepsol, macrogol, tween 61, cacao butter, laurin, glycerogelatin and the like.

The pharmaceutical composition of the present invention may be administered orally or parenterally, and it is preferable to select the intraperitoneal, rectal, rectal, intravenous, intramuscular, subcutaneous, intrauterine or intracerebral injection methods during parenteral administration, It is preferably used for external skin application.

The pharmaceutical composition according to the present invention is administered in a pharmaceutically effective amount. In the present invention, "pharmaceutically effective amount" means an amount sufficient to treat a disease at a reasonable benefit / risk ratio applicable to medical treatment, and the effective dose level will depend on the type of disease, severity, , Sensitivity to the drug, time of administration, route of administration and rate of release, duration of treatment, factors including co-administered drugs, and other factors well known in the medical arts. The pharmaceutical composition of the present invention can be administered as an individual therapeutic agent or in combination with other therapeutic agents, and can be administered sequentially or simultaneously with conventional therapeutic agents, and can be administered singly or in multiple doses. It is important to take into account all of the above factors and to administer the amount in which the maximum effect can be obtained in a minimal amount without side effects, which can be easily determined by those skilled in the art.

The dosage of the pharmaceutical composition of the present invention may be varied depending on the patient's body weight, age, sex, health condition, diet, administration time, administration method, excretion rate, and severity of the disease.

The present invention also provides a health functional food composition for preventing or ameliorating a melanin pigment hyperpigmentation disease or inflammatory disease, comprising a jujube extract prepared by a method comprising the steps of:

(a) preparing a jujuba seed powder by drying and pulverizing the jujube seeds;

(b) hydrolyzing the mixture of the jujube seed powder prepared in step (a) with water by adding an enzyme to the mixture;

(c) treating the hydrolyzate of step (b) with 100% ethanol; And

(d) homogenizing the ethanol-treated hydrolyzate of step (c) by ultra-high pressure.

When the health functional food composition of the present invention is used as a food additive, the health functional food composition may be added as it is, or may be used together with other food or food ingredients, and suitably used according to a conventional method. The amount of the active ingredient can be suitably used depending on its use purpose (prevention or improvement). Generally, the health functional food composition of the present invention is added in an amount of not more than 15 parts by weight, preferably not more than 10 parts by weight based on the raw material, when the food or beverage is produced. However, in case of long-term ingestion for health, the amount may be less than the above range, and since there is no problem in terms of safety, the active ingredient may be used in an amount exceeding the above range.

There is no particular limitation on the kind of the health functional food. Examples of the foods to which the health functional food composition can be added include dairy products including meat, sausage, bread, chocolate, candy, snacks, confectionery, pizza, ramen and other noodles, gums, ice cream, soups, Drinks, alcoholic beverages, and vitamin complexes, all of which include health foods in a conventional sense.

In addition, the health functional food composition of the present invention can be produced as a food, particularly a functional food. The functional food of the present invention includes components that are ordinarily added in food production, and includes, for example, proteins, carbohydrates, fats, nutrients, and seasonings. For example, in the case of a drink, a natural carbohydrate or a flavoring agent may be included as an additional ingredient in addition to the active ingredient. The natural carbohydrate may be selected from the group consisting of monosaccharides (e.g., glucose, fructose, etc.), disaccharides (e.g., maltose, sucrose etc.), oligosaccharides, polysaccharides (e.g., dextrin, cyclodextrin, , Xylitol, sorbitol, erythritol, etc.). The flavoring agent may be a natural flavoring agent (e.g., tau Martin, stevia extract, etc.) and a synthetic flavoring agent (e.g., saccharin, aspartame, etc.).

In addition to the above health functional food composition, it is also possible to use various nutrients, vitamins, electrolytes, flavors, colorants, pectic acids and salts thereof, alginic acid and salts thereof, organic acids, protective colloid thickening agents, pH adjusters, stabilizers, preservatives, glycerin, A carbonating agent used in beverages, and the like.

Hereinafter, embodiments of the present invention will be described in detail. However, the following examples are illustrative of the present invention, and the present invention is not limited to the following examples.

Manufacturing example  1. Production of Daechu Seed Extract

Extracts were prepared as follows according to enzyme type treatment, presence / absence of ultrahigh pressure homogenization, and enzyme treatment + hyperbaric homogenization in order to observe the extraction yield and compositional composition of the extracts according to the type of enzymes and the presence or absence of ultrahigh pressure homogenization.

(1) Enzyme treatment

30 ml of distilled water adjusted to pH 4.5 in 2.5 g of dried and pulverized jujube seed sample and 50 units / ml and 500 unit / ml of enzymes, Viscozyme and Pectinex, were added at 50 ° C and 37 ° C At 180 rpm. The enzyme was treated at the optimum time (10 hours and 2 hours), and then 70 ml of 100% ethanol was added. The mixture was extracted at 180 rpm for 2 hours at room temperature and then centrifuged at 12,000 rpm for 15 minutes.

(2) Ultra high pressure homogenization treatment

30 ml of distilled water adjusted to pH 4.5 and 70 ml of 100% ethanol were added to 2.5 g of the dried and pulverized jujuba seed sample, and the mixture was stirred for 12 minutes at 12,000 rpm for 15 minutes, filtered to prepare a filtrate, The pressure was passed twice through the orifice tube.

(3) Enzyme treatment and high pressure homogenization parallel treatment

30 ml of distilled water adjusted to pH 4.5 in 2.5 g of dried and pulverized jujube seed sample and 50 units / ml and 500 unit / ml of enzymes, Viscozyme and Pectinex, were added at 50 ° C and 37 ° C At 180 rpm. The enzymes were treated at optimal times (10 hours and 2 hours), and 70 ml of 100% ethanol was added. The mixture was extracted at 180 rpm for 2 hours at room temperature (25 ° C), centrifuged at 12,000 rpm for 15 minutes, And then passed through an orifice tube twice under a pressure of 120 MPa.

Example  1. Identification of extraction yield of jujube seeds

Changes in the extraction yields of various enzymes and ultrahigh - pressure treatments were measured for the functional materials using the extracts of Daechu seed. As a result, it was found that the extraction yield of the enzyme treatment was improved and the extraction yield of about 46% was improved in the case of the ultrahigh pressure homogenization. Especially, when the enzymatic treatment and the ultrahigh pressure homogenization treatment were combined, the yield enhancement effect was better than that of the jujube seed powder. Especially, the highest extraction yield was obtained by 100.7% increase of the ultra high pressure homogenization treatment after the treatment with Viscozyme One).

Extraction yield (%) of jujube seeds by enzyme treatment and ultra high pressure homogenization treatment Processing conditions of jujube seeds Extraction yield (%) Growth rate compared to untreated group (%) Untreated group 33.13 + 0.02 Enzyme V (viscozyme) 35.20 ± 2.86 6.2 Enzyme P (pectinex) 39.80 +/- 1.42 20.1 Ultrahigh pressure homogeneity 48.47 ± 1.25 46.3 Enzyme V + Ultra High Pressure Homogeneity 66.50 ± 0.68 100.7 Enzyme P + ultrahigh pressure homogeneity 51.02 + - 0.24 53.9

Example  2. Whitening efficacy of jujube extract

Example 2-1. Cytotoxicity measurement

In order to determine the concentration range to be used for the whitening efficacy test, the MTT (3- (4,5-dimeth-ylthiazol) -dihydroxybenzoate (MTT) -2-yl) -2,5-diphenyltetrazolium bromide) assay. B16F10 cells were cultured in 96-well plates with 100 μL of DMEM medium for 24 hours. 10 μL of 5 mg / mL MTT solution was added to each well, followed by incubation for 3 hours to induce a reduction reaction. 100 μL of dimethyl sulfoxide (DMSO) solution was added to completely dissolve the violet formazan crystals. Absorbance was measured at 570 nm using a spectrophotometer. The cytotoxicity was calculated as the relative cell viability of the drug-treated group based on the survival rate of 100% of the untreated cells cultured alone.

To investigate the cytotoxicity of jujube extract and hypertension and enzyme - treated extracts in B16F10 cells, MTT assay was carried out for 24 hours at a concentration of 0.4 mg / mL. When compared with the normal group treated with DMEM alone, cell viability exceeded 80% at all concentrations, indicating no cytotoxicity (Fig. 1 (A)). Therefore, the cells were treated with concentrations of 0.1, 0.2, and 0.4 mg / mL, which do not show cytotoxicity, and the whitening efficacy test was performed.

2-2. Melanin ( Melanin ) Biosynthesis and Tyrosinase ) Active measurement

Melanin biosynthesis was carried out by inoculating B16F10 cells into 12 wells at a density of 1 × 10 ⁵ cells / mL. After 24 hours, the jujube extract (0.1, 0.2, 0.4 mg / mL) and arbutin mu] g / mL) for 48 hours. After treatment, the cells were washed twice with phosphate buffered saline (PBS) and centrifuged to make cell precipitate. 200 μL of 1 N NaOH solution containing 10% dimethyl sulfoxide (DMSO) was added and dissolved at 60 ° C for 1 hour. Absorbance was measured at 405 nm. In order to measure tyrosinase inhibitory activity, 160 μL of a substrate solution in which 10 mM L-3,4-dihydroxypheny-lalanine (L-DOPA) was dissolved in 1 mL of a 0.1 M sodium phosphate buffer (pH 6.8) and 40 μL Was reacted at 37 캜 for 1 hour to measure the DOPA chrome produced in the reaction solution at 475 nm. Tyrosinase inhibitory activity was expressed by the absorbance reduction rate of the sample solution and the non-added sample.

As a result of measurement of melanin biosynthesis amount of jujube seed extract by enzyme treatment and homogenization process, when melanin-induced α-MSH group was regarded as melanin production 100%, no treatment group (jujube seed extract without enzyme treatment or ultra high pressure extraction) Melanin production was inhibited by about 40%. In the enzyme-treated group, the melanin production was inhibited by about 40% compared with the untreated group. However, the enzyme treatment did not show a significant effect compared with the untreated group. (Ar), which is a whitening agent, and exhibited higher efficacy than the arbutin (Ar), which is a whitening agent (FIG. 1 (B)).

Tyrosinase activity was similar to that of the previous melanin assay when the extract was treated at a concentration of 0.4 mg / mL. When skin is exposed to ultraviolet rays, it starts from tyrosinase, and after a series of oxidative polymerization reaction, melanin generated by skin causes spots and age-related erythema and promotes skin aging. The enzyme that plays an important role in this process is tyrosinase, and inhibition of tyrosinase enzyme is known to inhibit melanin biosynthesis. Therefore, it is considered that the jujube extract can be used as a whitening functional cosmetic material which can prevent skin pigmentation by inhibiting tyrosinase activity. (Ar), which is a whitening agent, and showed higher efficacy by reducing the activity of tyrosinase than that of untreated group in the combination of ultra high pressure homogenization treatment, enzyme treatment and ultrahigh pressure homogenization (Fig. 1 (C)).

Example  3. Anti-inflammatory efficacy of jujube extract

Example  3-1. Cytotoxicity measurement

In order to determine the concentration range to be used for measurement of NO (nitric oxide) production ability by examining the concentration on the cytotoxicity of the jujube extract or the ultrahigh pressure homogenate and the enzyme-treated extract, MTT (3- (4,5 -dimeth-ylthiazol-2-yl) -2,5-diphenyltetrazolium bromide) assay. RAW 264.7 cells were cultured in 96-well plates with 100 μL of DMEM medium for 24 hours. 10 μL of 5 mg / mL MTT solution was added to each well, followed by incubation for 3 hours to induce a reduction reaction. 100 μL of dimethyl sulfoxide (DMSO) solution was added to completely dissolve the violet formazan crystals. The degree of color development was measured at 570 nm using a spectrophotometer, and the relative cell viability of the drug-treated group was calculated based on the survival rate of 100% of the untreated group in which the cytotoxicity was only cultured.

As a result of measuring the cytotoxicity of the jujube extract, the jujube extract showed no cytotoxicity even at a concentration of 0.4 mg / mL. Therefore, the antiinflammatory effect of the jujube extract was measured at a concentration of 0.4 mg / mL, (Fig. 2 (A)).

Example  3-2. NO ( nitric oxide ) Generation  Measure

The nitric oxide (NO) concentration was measured by griess reagent. RAW 264.7 cells were adjusted to 1 × 10 ⁶ cells / mL using DMEM medium, and then inoculated on a 96-well plate and incubated for 24 hours at 37 ° C in a 5% CO ₂ incubator. 0.4 mg / mL of jujube extract or ultra-high pressure homogenized and enzyme-treated extract was treated for 1 hour, treated with 1 μg / mL of LPS and cultured for 24 hours. The supernatant of the culture solution was reacted with the griess reagent, and the absorbance at 540 nm was measured with a spectrophotometer to show the NO production rate as a percentage.

In the RAW264.7 cell induced by LPS, the NO concentration of the jujube extract and the hypertension and enzyme treated extracts were compared with those of the untreated group, the enzyme treated group, the ultrahigh pressure homogenized group, the enzyme treated group and the ultra high pressure homogenized group (Fig. 2 (B)). In particular, the combination of enzyme treatment and ultrahigh pressure homogenization showed the lowest production (Fig. 2 (B)).

Claims (8)

A cosmetic composition for skin whitening comprising as an active ingredient a jujube extract prepared by a method comprising the steps of:
(a) preparing a jujuba seed powder by drying and pulverizing the jujube seeds;
(b) adding viscozyme or pectinex to the mixture obtained by adding water to the jujuba seed powder prepared in step (a), and then hydrolyzing the mixture;
(c) treating the hydrolyzate of step (b) with 100% ethanol; And
(d) homogenizing the ethanol-treated hydrolyzate obtained in step (c) under a pressure of 100 to 120 MPa. delete delete The method according to claim 1, wherein the jujube seed extract
(a) preparing a jujuba seed powder by drying and pulverizing the jujube seeds;
(b) Viscozyme or Pectinex was added to a mixture prepared by adding 10 to 14 times (v / w) of water adjusted to pH 3.5 to 5.5 to the jujube seed powder prepared in step (a) Followed by hydrolysis at 30 to 60 ° C for 1 to 12 hours;
(c) adding 6 to 8 times (v / v) 100% ethanol to the hydrolyzate of step (b) and treating the mixture at 20 to 30 ° C for 1 to 3 hours; And
(d) homogenizing the ethanol-treated hydrolyzate of step (c) at a pressure of 100 to 120 MPa under ultrahigh pressure to prepare a cosmetic composition for skin whitening. A pharmaceutical composition for preventing or treating a melanin pigment hyperproliferative disorder, comprising an extract of a jujube produced by a method comprising the steps of:
(a) preparing a jujuba seed powder by drying and pulverizing the jujube seeds;
(b) adding viscozyme or pectinex to the mixture obtained by adding water to the jujuba seed powder prepared in step (a), and then hydrolyzing the mixture;
(c) treating the hydrolyzate of step (b) with 100% ethanol; And
(d) homogenizing the ethanol-treated hydrolyzate obtained in step (c) under a pressure of 100 to 120 MPa. A health functional food composition for preventing or ameliorating a melanin pigment hyperpigmentation disease comprising an extract of a jujube produced by a method comprising the following steps as an active ingredient:
(a) preparing a jujuba seed powder by drying and pulverizing the jujube seeds;
(b) adding viscozyme or pectinex to the mixture obtained by adding water to the jujuba seed powder prepared in step (a), and then hydrolyzing the mixture;
(c) treating the hydrolyzate of step (b) with 100% ethanol; And
(d) homogenizing the ethanol-treated hydrolyzate obtained in step (c) under a pressure of 100 to 120 MPa. A pharmaceutical composition for the prophylaxis or treatment of inflammatory diseases, comprising an extract of a jujube produced by a method comprising the following steps as an active ingredient:
(a) preparing a jujuba seed powder by drying and pulverizing the jujube seeds;
(b) adding viscozyme or pectinex to the mixture obtained by adding water to the jujuba seed powder prepared in step (a), and then hydrolyzing the mixture;
(c) treating the hydrolyzate of step (b) with 100% ethanol; And
(d) homogenizing the ethanol-treated hydrolyzate obtained in step (c) under a pressure of 100 to 120 MPa. A health functional food composition for preventing or ameliorating an inflammatory disease comprising as an active ingredient a jujube extract prepared by a method comprising the steps of:
(a) preparing a jujuba seed powder by drying and pulverizing the jujube seeds;
(b) adding viscozyme or pectinex to the mixture obtained by adding water to the jujuba seed powder prepared in step (a), and then hydrolyzing the mixture;
(c) treating the hydrolyzate of step (b) with 100% ethanol; And
(d) homogenizing the ethanol-treated hydrolyzate obtained in step (c) under a pressure of 100 to 120 MPa.

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