KR20110096784A - Composition for preventing and treating inflammation containing essential oil from a seed of ziziphus jujuba - Google Patents
Composition for preventing and treating inflammation containing essential oil from a seed of ziziphus jujuba Download PDFInfo
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- KR20110096784A KR20110096784A KR1020100016225A KR20100016225A KR20110096784A KR 20110096784 A KR20110096784 A KR 20110096784A KR 1020100016225 A KR1020100016225 A KR 1020100016225A KR 20100016225 A KR20100016225 A KR 20100016225A KR 20110096784 A KR20110096784 A KR 20110096784A
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- essential oil
- jujube
- tpa
- inflammation
- seed
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- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K8/00—Cosmetics or similar toiletry preparations
- A61K8/18—Cosmetics or similar toiletry preparations characterised by the composition
- A61K8/96—Cosmetics or similar toiletry preparations characterised by the composition containing materials, or derivatives thereof of undetermined constitution
- A61K8/97—Cosmetics or similar toiletry preparations characterised by the composition containing materials, or derivatives thereof of undetermined constitution from algae, fungi, lichens or plants; from derivatives thereof
- A61K8/9783—Angiosperms [Magnoliophyta]
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Abstract
The present invention relates to a pharmaceutical composition for preventing and treating inflammation and a nutraceutical composition containing an essential oil derived from jujube seed as an active ingredient, wherein the jujube seed essential oil according to the present invention is potent in a short period of time in an animal model of TPA-induced skin inflammation. It has been shown to inhibit skin inflammation and effectively inhibit inflammatory responses in animal models of chronic skin inflammation.
Description
The present invention relates to a composition for preventing and treating inflammation containing jujube seed-derived essential oil, and more particularly, to a pharmaceutical composition and health functional food composition for preventing and treating inflammation containing jujube seed-derived essential oil as an active ingredient.
Inflammation is the body's physiological response to stimuli, including infection and tissue damage. Excessive and persistent inflammation, however, causes various pathological symptoms, such as bacterial pulmonary disease, rheumatoid arthritis and skin inflammation. The skin is the primary point of contact between the body and the external environment, providing a primary line of defense against traumatic symptoms and invasion by microbial pathogens. The skin also has many active defense mechanisms in addition to its properties as a physical barrier, and the regulation of this mechanism is important because inappropriate or mistargeted immune activity is involved in the pathogenesis of a wide range of inflammatory skin diseases. Some of these symptoms are readily treated, but the treatment of chronic inflammatory diseases such as psoriasis and atopic dermatitis is not 100% successful.
High levels of inflammatory cytokines and reactive oxygen species are believed to be responsible for the pathophysiological mechanisms associated with various inflammatory skin diseases. It is well known that a variety of inflammatory cell populations interact and diffuse to the site of inflammation in response to the release of soluble proinflammatory mediators such as cytokines, prostaglandins and leukotrienes, thereby inducing and maintaining skin inflammation.
Recent therapies have focused on treating the symptoms of skin diseases by combining wetting agents, antihistamines, antibiotics and corticosteroids with the aim of regenerating barrier function and relieving itching, secondary infections and inflammation. However, steroids can destroy multiple cytokine networks involved in lymphocyte function, resulting in immunosuppression, and can lead to skin atopy by lowering collagen synthesis over long-term topical use. Because of this risk, new therapeutic approaches are being intensively studied.
Inflammatory diseases are currently being treated with steroidal and nonsteroidal anti-inflammatory drugs (NASAIDs). Unfortunately, these widely formulated groups of drugs have significant negative side effects, reducing the use of certain classes of people. Therefore, there is a need to develop new drugs with new mechanisms of action that do not bring significant side effects. Natural product based anti-inflammatory agents with good efficacy and low side effects risk are believed to provide promising therapeutic and prophylactic effects of inflammation related symptoms.
Many plant species contain a variety of bioactive compounds that exhibit health beneficial properties, antioxidant, anti-inflammatory and antimicrobial effects, and their therapeutic use is also increasing. Various natural products have already been tested in many animal models to develop new anti-inflammatory therapies.
Jujube fruit is described as the "fruit of life," and the ancient Chinese have known his unique characteristics for thousands of years. Today practitioners are looking for scientific evidence of the special properties of jujube. Jujube (jujube or red date) is a kind of scientific name Ziziphus jujuba and is a thorny rhamnaceous plant of the buckthorn family Rhamnaceae, and mainly uses the fruit. The exact natural distribution of jujube is not clear due to its extensive cultivation, but it is considered to be South Asia, Syria, North India, South China and Central China, and according to the Wikipedia Encyclopedia, it is also distributed in southeast Europe. Jujube fruits are edible and various parts of the jujube ( Ziziphus jujuba ) have multiple pharmacological properties, such as contraception, analgesic and antidiabetic properties. In some regions, stem mixtures of jujube ( Ziziphus jujuba ) are used to prevent pregnancy. However, little scientific research has been conducted on the effects of Ziziphus jujuba seeds. In traditional medicine, the seeds of jujube ( Ziziphus jujuba ) are used to relieve insomnia and anxiety. According to the conventional report, Ziziphus jujuba seeds have been reported to be effective in improving the lipid composition in blood glucose, serum of dietary hyperlipidemic rats. In particular, Ziziphus jujuba seeds were more effective as a therapeutic diet for controlling metabolic disorders in adult diseases. However, there are no reports of anti-inflammatory activity of essential oils derived from jujube ( Ziziphus jujuba ).
Accordingly, the present inventors have conducted extensive research to develop an anti-inflammatory agent using natural products, and as a result, confirm that the essential oil derived from jujube ( Ziziphus jujuba ) seed has anti-inflammatory activity and confirm the mechanism of the anti-inflammatory effect. Was done.
Accordingly, an object of the present invention is to provide a pharmaceutical composition for preventing and treating inflammation containing essential oil derived from jujube ( Ziziphus jujuba ).
Another object of the present invention is to provide a dietary supplement for preventing and improving inflammation containing essential oil derived from jujube ( Ziziphus jujuba ) seed.
Another object of the present invention is to provide a cosmetic composition for preventing and improving inflammation containing essential oil derived from jujube ( Ziziphus jujuba ) seed.
The purpose of the present invention as described above is to isolate the essential oil from jujube seed, TPA-induced inflammation analysis and mouse ear tissue morphology analysis in mice using the separated jujube seed essential oil TPA-induced skin inflammation It was achieved by demonstrating strong skin inflammation inhibitory effect in a short period of time in animal models and effective inhibition of inflammatory responses in animal models of chronic skin inflammation.
The present invention provides a pharmaceutical composition for preventing and treating inflammation containing the essential oil derived from jujube ( Ziziphus jujuba ) seed as an active ingredient.
In another aspect, the present invention provides a dietary supplement for preventing and improving inflammation containing the essential oil derived from jujube ( Ziziphus jujuba ) seed as an active ingredient.
The present invention also provides a cosmetic composition for preventing and improving inflammation containing the essential oil derived from jujube ( Ziziphus jujuba ) seed as an active ingredient.
In the present invention, the term "active ingredient" refers to a substance or a group of substances (including medicinal herbs whose pharmacologically active ingredients are not known) which are expected to express directly or indirectly the efficacy effect of the medicine by inherent pharmacological action. It means containing a main component as.
In the present invention, "health functional food" means a food manufactured and processed using raw materials or ingredients having functional properties useful for the human body according to the Health Functional Food Act No. 6767, and "functional" means a human body It means the ingestion for the purpose of obtaining a useful effect in health use such as nutrient control or physiological action on the structure and function of.
Pharmaceutical compositions comprising the jujube seed essential oil of the present invention may further comprise suitable carriers, excipients and diluents commonly used in the manufacture of pharmaceutical compositions.
The pharmaceutical composition comprising the essential oil of jujube seed of the present invention may be prepared in the form of powders, granules, tablets, capsules, suspensions, emulsions, syrups, aerosols, oral formulations, external preparations, suppositories, and sterile injectable solutions, respectively. It can be formulated and used in the form.
Carriers, excipients and diluents that may be included in the pharmaceutical composition comprising the jujube seed essential oil of the present invention include lactose, dextrose, sucrose, sorbitol, mannitol, xylitol, erythritol, maltitol, starch, acacia rubber, alginate, Gelatin, calcium phosphate, calcium silicate, cellulose, methylcellulose, microcrystalline cellulose, polyvinyl pyrrolidone, water, methylhydroxybenzoate, propylhydroxybenzoate, talc, magnesium stearate and mineral oil. When formulated, diluents or excipients such as fillers, extenders, binders, wetting agents, disintegrating agents, and surfactants are usually used. Solid preparations for oral administration include tablets, pills, powders, granules, capsules, and the like, and the solid preparations include at least one excipient such as starch, calcium carbonate, sucrose in the extract. Or lactose, gelatin, or the like is mixed. In addition to simple excipients, lubricants such as magnesium stearate and talc are also used. Oral liquid preparations include suspensions, solvents, emulsions, and syrups, and may include various excipients, such as wetting agents, sweeteners, fragrances, and preservatives, in addition to commonly used simple diluents such as water and liquid paraffin. . Formulations for parenteral administration include sterile aqueous solutions, non-aqueous solvents, suspensions, emulsions, lyophilized preparations, suppositories. As the non-aqueous solvent and suspending agent, propylene glycol, polyethylene glycol, vegetable oil such as olive oil, injectable ester such as ethyl oleate and the like can be used. As the base of the suppository, witepsol, macrogol, tween 61, cacao butter, laurin butter, glycerogelatin and the like can be used.
The preferred dosage of the jujube seed essential oil according to the present invention depends on the condition and weight of the patient, the extent of the disease, the form of the drug, the route of administration and the duration, and may be appropriately selected by those skilled in the art. However, for the preferred effect, the jujube seed essential oil of the present invention is preferably administered at 0.0001 to 100 mg / kg, preferably at 0.001 to 100 mg / kg. Administration may be administered once a day or may be divided several times. The dosage does not limit the scope of the invention in any aspect.
The jujube seed essential oil according to the present invention can be administered to mammals such as mice, mice, livestock, humans, etc. by various routes. All modes of administration can be expected, for example, oral, rectal or intravenous, intramuscular, subcutaneous, intrauterine dural or intracerebroventricular injections and directly to the skin.
In addition, the jujube seed essential oil according to the present invention may be added to food or beverage for the purpose of preventing and improving inflammation. Examples of foods to which jujube seed essential oil can be added include various foods, beverages, gums, teas, vitamin complexes, health functional foods, powders, granules, tablets, capsules or beverages. At this time, the amount of the jujube seed essential oil in the food or beverage may be added at 0.01 to 15% by weight of the total food weight, the health beverage composition is 0.02 to 5 g, preferably 0.3 to 1 g based on 100 ml Can be added.
The food composition of the present invention is not particularly limited to other ingredients except jujube seed essential oil as essential ingredients in the indicated ratios, and may contain various flavors or natural carbohydrates as additional ingredients, such as ordinary drinks. Examples of the above-mentioned natural carbohydrates include monosaccharides such as glucose, fructose and the like; Disaccharides such as maltose, sucrose and the like; And conventional sugars such as polysaccharides such as dextrin, cyclodextrin, and sugar alcohols such as xylitol, sorbitol, and erythritol. As flavoring agents other than those described above, natural flavoring agents (tauumatin, stevia extract (e.g., Rebaudioside A, glycyrrhizin, etc.) and synthetic flavoring agents (saccharin, aspartame, etc.) can be advantageously used. The proportion of said natural carbohydrates is generally about 1-20 g, preferably about 5-12 g per 100 ml of the composition of the present invention.
In addition to the above, the jujube seed essential oil of the present invention includes various nutrients, vitamins, minerals (electrolytes), flavors such as synthetic flavors and natural flavors, coloring and neutralizing agents (such as cheese and chocolate), pectic acid and salts thereof, alginic acid And salts thereof, organic acids, protective colloid thickeners, pH adjusters, stabilizers, preservatives, glycerin, alcohols, carbonation agents used in carbonated beverages, and the like. In addition, the jujube seed essential oil of the present invention may contain a pulp for producing natural fruit juice and fruit juice beverage and vegetable beverage. These components can be used independently or in combination. The proportion of such additives is not so critical, but the jujube seed essential oil of the present invention is generally selected in the range of 0 to about 20 parts by weight per 100 parts by weight.
In the cosmetic composition of the present invention, the composition is characterized in that the formulation of the lotion (skin lotion), lotion, cream, massage cream, essence, pack. The formulations may be prepared using methods known in the art. In addition, the content of jujube seed essential oil in the formulation may also be included in the content range (about 1-10% by weight) conventionally known in the art.
Components included in the cosmetic composition of the present invention may include components commonly used in cosmetic compositions in addition to the jujube seed essential oil as an active ingredient, for example, such as stabilizers, solubilizers, vitamins, pigments and flavorings Phosphorus aids and carriers.
In the present invention, jujube seed was steam distilled to obtain a pale yellow oil mainly composed of phenolic compounds, oxygenated mono- and sesquiterpenes, and oils having their respective hydrocarbons.
In the present invention, to evaluate the combined effect of the jujube seed essential oil on TPA-induced skin inflammation, the effect of jujube essential oil on TPA_induced skin inflammation was investigated in a dose dependent manner. As a preliminary step, the present invention used 1% and 10% essential oils for in vivo experiments, which resulted in almost equally reduced TPA-induced ear thickness and skin moisture content. Based on the in vivo results, only 1% essential oil was used in histological analysis.
As a result, the present invention revealed that the inhibitory effect of jujube seed essential oil on TPA-induced skin inflammation was superior to eugenol (see FIGS. 2E and 2D). It should be noted that hydrocortisone, a positive control, did not show a significant effect in the ears of TPA-induced mice (see FIG. 2C). Jujube seed essential oil according to the present invention showed a strong anti-inflammatory effect in a very short period of time compared to hydrocortisone (HC) (positive control) currently prescribed for the treatment of various inflammatory skin diseases.
The present invention clearly demonstrated that jujube seed essential oil inhibited the inflammatory response in animal models of chronic skin inflammation. In addition, in the present invention, in order to further investigate the anti-inflammatory effect of jujube seed essential oil, the thickness of the dermis and the cortical area of the ear tissue was measured using an ear biopsy. As a result, it was found that the jujube seed essential oil significantly inhibited the increase in ear thickness due to TPA induction in both the skin and dermal areas compared to the control (see FIGS. 3 and 4).
Thus, histological analysis clearly shows that jujube essential oil may be beneficial as an excellent therapeutic agent for the treatment of various inflammatory diseases.
As described above, the jujube seed essential oil according to the present invention exhibited a strong skin inflammation inhibitory effect in a short period of time in the TPA-induced skin inflammation animal model, and effectively inhibited the inflammatory response in the chronic skin inflammation animal model. It can be used as an anti-inflammatory and is expected to be used for the development of new drugs for various inflammatory diseases.
1 is a graph showing the effect of jujube seed essential oil on TPA-induced ear thickness and moisture content in BLAB / c mice.
Figure 2 is a diagram showing the histological cross section of a mouse ear skin biopsy showing the epidermis, dermis and cartilage layer of the mouse. A: no treatment, B: TPA treatment, C: HC treatment, D: eugenol treatment, E: jujube seed essential oil treatment
3 is a graph showing the area measurement results for evaluating the thickness of the mouse epidermis.
Figure 4 is a graph showing the area measurement results for evaluating the thickening of the mouse dermis.
Hereinafter, preferred embodiments of the present invention will be described in more detail with reference to Examples. However, the scope of the present invention is not limited to these examples.
Example
Jujube seeds were collected in August 2008 in Gyeongsan, Korea, washed, dried and ground.
TPA (12-O-tetradecanoylphorbol-13-acetate) and hydrocortisone were purchased from Sigma.Aldrich (St. Louis, MO, USA), and all other compounds and reagents used commercial grades. It was.
As experimental animals, 5-week-old female BALB / c mice (18-20 g) were purchased from Orient Bio, Inc. (Seoul, South Korea), and placed in polypropylene cages (3 per cage) in a standard laboratory. Diet and water were optionally fed. The experimental animals were bred for about 7 days before the experiment in a 12:12 hour contrast cycle in an air-conditioned room. Room temperature (about 23 ° C.) and humidity (about 60%) were automatically adjusted.
The results of all experiments are expressed as mean ± SD. One-way ANOVA and Dennetts t-test were used for multiple comparisons using GraphPad Prism (GraphPad Software, Inc., San Diego, Calif., USA). The statistical significance criterion was p <0.05.
Example 1 Separation of Jujube Seed Essential Oil
Approximately 250 g of crushed jujube seeds were steam distilled for 3 hours using a Klevenzer-type apparatus to obtain an essential oil as a pale yellow oil mainly composed of oils having phenolic compounds, oxygenated mono- and sesquiterpenes and their respective hydrocarbons. . The obtained essential oil was dried over anhydrous Na 2 SO 4 and stored at 4 ° C. in a sealed vial until further analysis.
Experimental Example 1 Analysis of TPA-Induced Inflammation in Mice
In this experimental example, a powerful anti-inflammatory effect of jujube seed essential oil treatment was analyzed using a TPA-induced skin inflammation model to test whether jujube essential oil can alleviate skin inflammation.
TPA (12-O-tetradecanoylphorbol-13-acetate) was used to induce skin inflammation in BALB / c mice, increasing ear thickness and skin moisture content.
TPA was dissolved in AOO (acetone: olive oil = 4: 1) and used as a skin inflammation inducer. To induce skin inflammation in mice, 10 μL of said inflammation inducer was added to the inner and outer epidermis, respectively. Thirty minutes prior to TPA treatment, 10 μL of sample solution, its excipients as a control, were topically treated. Hydrocortisone (HC), which is currently used to treat various inflammatory skin diseases, was used as a positive control. Ear thickness was measured with a hand-held thickness meter in the range 0-9 mm, modified to increase the contact surface area to reduce the load on the ear tip at 0.01 mm intervals. Ear thicknesses were measured before TPA treatment and 4 hours after TPA treatment (b = TPA alone; b ′ = TPA + sample). The following values were then calculated:
Edema A induced by TPA alone (b-a);
Edema B induced by TPA + sample (b'-a);
% Inhibition = [(edema A-edema B) / edema A] × 100
After measuring ear thickness, mice were anesthetized, 6 mm 2 diameter ear punch biopsies were collected and weighed with a Mettler-Toledo (AE-163) electronic balance, respectively, and the weighed weight was wet weight. It was defined as (Wet weight). The ear biopsy was dried in a drying oven for 24 hours, weighed again, and the weighed weight was defined as the dry weight. The dry weight was subtracted from the wet weight and divided by the dry weight to calculate the skin moisture content and expressed as mg H 2 O / mg dry weight.
Increased skin thickness increase is often the primary feature of skin irritation and local inflammation. This parameter is an indicator of the number of processes that occur during skin inflammation, including enhanced vascular permeability, edema and swelling in the dermis and proliferation of epidermal keratinocytes. Exposure of the ears of BALB / c mice to TPA significantly increased skin thickness (FIG. 1A) and skin moisture content (FIG. B). As shown in FIG. 1A, TPA (4 mM) substantially increased ear thickness, while local treatment of jujube seed essential oil significantly inhibited the increase in ear thickness. When ear thickness was measured, a two-fold increase in TPA-induced mouse ears (0.54 ± 0.04 mm) was observed compared to untreated mice (0.23 ± 0.01 mm). 1% and 10% jujube seed essential oil treatments resulted in a significant decrease in ear thickness, measured 0.3 ± 0.02 and 0.35 ± 0.01 mm, respectively. The effect of 1% and 10% jujube seed essential oil on TPA-induced mouse ears was also comparable to the effect of 1% HC (0.5 ± 0.1 mm) used as a positive control.
Increasing the water content induced by TPA was also inhibited in a concentration dependent manner by jujube seed essential oil treatment (see FIG. 1B). Treatment of 1% and 10% essential oils in the TPA-induced skin inflammation model resulted in a water content reduction of about 51 and 53%, respectively. However, hydrocortisone treatment in TPA-induced mouse ears resulted in a water content reduction of about 39%. As shown in Figures 1a and 1b, the inhibitory effect of 1% and 10% jujube seed essential oil on TPA-induced skin inflammation was better than in 1% HC. Therefore, these results show that jujube seed essential oil has a potent in vivo anti-inflammatory effect.
Experimental Example 2 Analysis of Morphology of Mouse Ear Tissue
For morphological analysis of skin inflammation, biopsies of the ears of control and each treatment mice were collected, fixed in 4% paraformaldehyde (0.1M phosphate buffer, pH 7.4) and decalcified. The immobilized tissue was sliced in succession thin to 5.0 μm using a microtome (LEICA RM 2125RT, 14 Nussloch, Germany). The cut sections were stained with Harrys hematoxylin-eosin, length was measured using light microscopy, and sample areas were selected for quantitative microscopic analysis of cell mediated inflammatory responses.
Examination of H & E-stained ear sections derived from TPA induced mice showed significant ear thickness increase as evidence of subsequent inflammatory cell infiltration with edema, epidermal proliferation, and connective tissue destruction in the skin with TPA treatment (FIG. 2A and FIG. 2b). Through histological comparison, 1% essential oil treatment reduced ear thickness and the level of associated pathological indicators to a degree comparable to eugenol and positive control hydrocortisone (HC) (see FIGS. 2C, 2D and 2E). . These results directly indicate the effect of jujube seed essential oil in the target tissue, providing additional evidence that jujube seed essential oil alleviates TAP-induced contact dermatitis.
To further elucidate the effect of jujube seed essential oil on TPA-induced skin inflammation, the skin and dermal area thicknesses of the mouse ear tissue were measured, and as shown in FIG. ± 2.9 μm 2 ) were nearly identical to controls (12.3 ± 2.1 μm 2 ), but the envelope area thicknesses for TPA, HC and eugenol were 19.9 ± 3.7, 17.8 ± 3.6 and 16.2 ± 3.1 μm 2, respectively.
Moreover, the dermal region thickness of the jujube essential oil treated ear tissue provided evidence that jujube essential oil had the greatest anti-inflammatory activity among all test samples (see FIG. 4). The dermal area thicknesses for the jujube seed essential oil, TPA, HC and eugenol were 185.5 ± 20.4, 249.7 ± 40.1, 309.7 ± 28.6 and 204.8 ± 28.2 μm 2, respectively.
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