Anti-inflammatory composition containing plum juice as an active ingredient

Abstract

The present invention relates to an anti-inflammatory composition containing ume squeezed juice as an active ingredient, and the feed composition is further added with aogiri extract, false acacia extract, butterbur extract, cinnamon extract and asparagus extract, so that it is harmless to the human body and has excellent efficacy in suppressing various inflammatory symptoms. Therefore, it can be used as a therapeutic agent for various inflammatory diseases such as colitis, arthritis, hepatitis, cystitis, nephritis, dermatitis, a food for improving symptoms, a cosmetic, and the like.

Description

[Technical Field]

[0001] The present invention relates to an anti-inflammatory composition containing plum squeezed juice as an active ingredient. [Background Art]

[0002] The mechanism by which an inflammatory reaction occurs is as follows: macrophages respond to pathogens to produce pro-inflammatory cytokines such as tumor necrotic factor-α (TNF-α), interleukin-6 (IL-6), and IL-1β, then synthesize inducible nitric oxide synthase (iNOS) and cyclooxygenase-2 (COX-2) to produce nitric oxide (NO) and prostaglandin E2 (PGE2). Physiologically, NO plays multiple roles including eliminating bacteria and tumors, regulating blood pressure, and mediating neurotransmission. However, when an inflammatory reaction occurs, the expression of iNOS in related cells increases to produce a large amount of NO; excessively produced NO induces tissue damage, gene mutation, nerve damage and the like, and increases vascular permeability to promote inflammatory reactions such as edema. PGE2 is an inflammatory factor mainly involved in pain and fever, and is produced by COX-2 in macrophages when an inflammatory reaction occurs. Therefore, among substances produced in an inflammatory reaction, the anti-inflammatory effect can be confirmed by verifying the suppression of production of substances such as NO and PGE2.

[0003] On the other hand, the activation process of macrophages by LPS is mediated by TLRs (toll-like receptors) and occurs via a pathway that activates the transcription factor NF-κB (nuclear factor-κB). LPS is present in the extracellular wall of Gram-negative bacteria and consists of a polysaccharide region comprising a core region and an O-polysaccharide region, and a lipid region known as lipid A. Lipid A of LPS is known to play an important role in the pathophysiology of Gram-negative bacteria. When TLR-4 on the surface of macrophages is stimulated by lipid A of LPS, a signal is transmitted to the TOLL / IL-1R (TIR) ​​domain in the cytoplasm. The signal transmitted in this way within the macrophage induces linkage with MyD88 (myeloid differentiation primary-response protein 88) and guides IRAK4 (IL-1R-associated kinase 4) into the signaling pathway. IRAK-4 phosphorylates IRAK1, and TRAF6 (tumor-necrosis-factor-receptor-associated factor 6) also joins the signaling pathway. Phosphorylated IRAK1 and TRAF6 separate from the receptor complex and, via other mechanisms at the cell membrane, reactivate TAK1 (transforming-growth-factor-β-activated kinase) in the cytoplasm. Activated TAK1 phosphorylates the MAP (mitogen-activated protein) kinase and IKK (inhibitor of nuclear factor-κB (IκB)-kinase complex) complex, inducing the degradation of IκB and NF-κB. The separated NF-κB then migrates to the macrophage nucleus, inducing the expression of inflammation-related genes and generating various cytokines and proteins.

[0004] Plums have long been used to prevent and treat food poisoning and infectious diseases. Even today, they are extensively studied for purposes such as food preservation, and their antibacterial properties, particularly due to their high content of citric acid and malic acid, are widely known.

[0005] While conducting various studies on anti-inflammatory compositions using such plums, the inventors discovered that the aforementioned plum juice suppresses NO production in macrophages activated by LPS, thus completing the present invention. [Prior art documents] [Patent Documents]

[0006] [Patent Document 1] Korean Published Patent Publication No. 10-2005-0117975 (Title of Invention: Natural Antioxidant Composition Containing Chinese Parasol Tree Extract and Method for Producing the Same, Applicant: Kim Jin-soo, Publication Date: December 15, 2005) [Patent Document 2] Korean Patent Publication No. 1384410 (Title of Invention: Application in the Production of a Plum Extract Composition, Applicant: Hanzo U-Mate Science and Technique Company, Limited et al., April 4, 2014) [Patent Document 3] Korean Patent Publication No. 10-2141623 (Title of Invention: Composition for the Prevention or Treatment of Inflammatory Pain Containing Cinnamon Extract as an Active Ingredient, Applicant: Dong-Eui University Industry-Academia Cooperation Group, Registration Date: December 20, 2018) [Patent Document 4] Korean Patent Publication No. 10-2141623 (Title of Invention: Anti-inflammatory active compound isolated from butterbur extract and method for separating the same, Applicant: NatureBio Co., Ltd., Registration Date: September 19, 2019) [Patent Document 5] Korean Patent Publication No. 10-1836679 (Title of Invention: Pharmaceutical Composition for the Prevention or Treatment of IL-1β-Related Diseases Containing Black Locust Leaf Extract as an Active Ingredient, Applicant: NatureBio Co., Ltd., Registration Date: March 2, 2018) [Overview of the project] [Problems that the invention aims to solve]

[0007] The object of the present invention is to provide an anti-inflammatory composition containing plum juice as an active ingredient. [Means for solving the problem]

[0008] The present invention relates to an anti-inflammatory composition characterized by containing plum juice, Chinese parasol tree extract, black locust extract, butterbur extract, cinnamon extract, and asparagus extract.

[0009] The anti-inflammatory composition may contain, per 100 parts by weight of plum juice, 20 to 30 parts by weight of Chinese parasol extract, 5 to 14 parts by weight of black locust extract, 20 to 30 parts by weight of butterbur extract, 3 to 15 parts by weight of cinnamon extract, and 10 to 20 parts by weight of asparagus extract.

[0010] The anti-inflammatory composition may contain a plum sugar extract. The plum sugar extract can be mixed in an amount of 10 to 30 parts by weight per 100 parts by weight of plum juice.

[0011] Each of the aforementioned extracts can be obtained by preparing each raw material sample and extracting it using water, C1-C4 alcohols, or a mixture thereof as a solvent. Water is preferably used as the solvent.

[0012] The present invention will be described in detail below. The plum juice used in this invention is characterized by being obtained by crushing fresh plums, pressing (juicing) them, and then heating the resulting liquid phase. Yellow plums or green plums can be used. Alternatively, the liquid phase obtained by crushing and pressing fresh plums can be heated once, centrifuged, filtered through a microfilter, and then heated again. Centrifugation is preferably performed at 4-10°C and 8000-10000 rpm for 5-30 minutes, and primary or secondary heating can be performed at 70-125°C for 0.5-48 hours. A microfilter with a pore size of 0.2-0.45 μm can be used.

[0013] The composition of the present invention may further contain a plum sugar extract. The plum sugar extract used in the present invention may be a liquid phase obtained by removing dregs from a mixture of plums and sugars that has been allowed to stand and extracted. The plum sugar extract may be a liquid phase obtained by removing dregs from a mixture of 50 to 200 parts by weight of sugars per 100 parts by weight of plums that has been allowed to stand and extracted for 3 to 6 months. The extraction temperature of the plum sugar extract is preferably 4 to 37°C, and more preferably 15 to 30°C. Any sugar that provides sweetness can be used, but one or more selected from the group consisting of sugar, sugar powder, oligosaccharides, agave syrup, maple syrup, cactus sugar, and honey can be used. In this case, if the amount of sugars mixed is less than 50 parts by weight per 100 parts by weight of plums, it is likely to spoil without fermentation. Furthermore, if sugars are added in excess of 200 parts by weight per 100 parts by weight of plums, the sugars may not mix properly with the juice extracted from the plums due to the excessive addition of sugar components. Also, if the extraction period is less than 3 months, spoilage is likely to occur thereafter, and if the extraction period exceeds 6 months, acetic acid fermentation or alcoholic fermentation may proceed, and spoilage may progress depending on the storage location or environment.

[0014] The Chinese parasol tree used in this invention may contain one or more species selected from branches, fruits, and leaves, and the woody part of the black locust (acacia) can be used.

[0015] Each solvent extract in the aforementioned anti-inflammatory composition can be mixed with plum juice after freeze-drying. However, if the extract is extracted with water, it may be mixed immediately with the plum juice. Preferably, the solvent extract may be the liquid phase obtained by adding 5 to 20 parts by weight of water to the raw material sample, heating at 70 to 90°C for 1 to 10 hours for extraction, and then removing the residue.

[0016] The solvent extracts used in this invention, namely Fiddlehead fern extract, black locust extract, butterbur extract, cinnamon extract, and asparagus extract, can be produced by the following method. For this purpose, each raw material sample is prepared, and the raw material sample can be extracted using water, C1-C4 alcohols, or a mixture thereof as a solvent. The C1-C4 alcohols can be selected from the group consisting of methanol, ethanol, propanol, isopropanol, butanol, and isobutanol.

[0017] The extraction conditions for the raw material sample may be 20-100°C for 1 minute to 48 hours. The process can be repeated 1 to 4 times. Conventional extraction equipment, ultrasonic grinding extractors, or fractionators can be used as extraction equipment. The solvent extract thus produced can be dried to remove the solvent by hot air drying, vacuum drying, or freeze-drying. The solvent extract can also be purified using column chromatography. The solvent extract can be fractionated or purified using conventional methods, such as extraction with organic solvents (alcohols, ethers, acetone, etc.), partitioning of hexane and water, or column chromatography, either individually or in a suitable combination. The chromatography can be selected from silica gel column chromatography, LH-20 column chromatography, ion exchange resin chromatography, medium pressure liquid chromatography, thin layer chromatography (TLC), silica gel vacuum liquid chromatography, and high performance liquid chromatography.

[0018] The present invention relates to a pharmaceutical composition for the prevention or treatment of inflammatory diseases, comprising the anti-inflammatory composition. The aforementioned inflammatory diseases may be selected from the group consisting of atopic dermatitis, edema, dermatitis, allergy, asthma, conjunctivitis, periodontitis, rhinitis, otitis media, systemic lupus erythematosus, fibromyalgia, psoriatic arthritis, osteoarthritis, rheumatoid arthritis, periarthritis of the shoulder, tendinitis, tenosynovitis, myositis, hepatitis, cystitis, nephritis, Sjögren's syndrome, and multiple sclerosis.

[0019] The aforementioned pharmaceutical compositions can be prepared by conventional methods in the form of oral dosage forms such as powders, granules, tablets, capsules, suspensions, emulsions, syrups, and aerosols, as well as topical preparations, suppositories, and sterile injection solutions. Examples of carriers, excipients, and diluents that may be included in the pharmaceutical compositions include lactose, dextrose, sucrose, sorbitol, mannitol, xylitol, erythritol, maltitol, starch, acacia gum, alginate, gelatin, calcium phosphate, calcium silicate, cellulose, methylcellulose, microcrystalline cellulose, polyvinylpyrrolidone, water, methyl hydroxybenzoate, propyl hydroxybenzoate, talc, magnesium stearate, and mineral oil. When formulated, they are prepared using commonly used fillers, bulking agents, binders, wetting agents, disintegrants, surfactants, and other diluents or excipients. Solid formulations for oral administration include tablets, pills, powders, granules, and capsules. Such solid formulations are prepared by mixing the anti-inflammatory composition of the present invention with at least one excipient, such as starch, calcium carbonate, sucrose or lactose, or gelatin. In addition to simple excipients, lubricants such as magnesium stearate and talc are also used. Liquid formulations for oral administration include suspensions, liquid formulations, emulsions, and syrups. In addition to commonly used simple diluents such as water and liquid paraffin, various excipients such as humectants, sweeteners, fragrances, and preservatives may be used. Formulations for parenteral administration include sterile aqueous solutions, non-aqueous solvents, suspensions, emulsions, lyophilized formulations, and suppositories. Non-aqueous solvents and suspensions that can be used include propylene glycol, polyethylene glycol, vegetable oils such as olive oil, and injectable esters such as ethyl oleate. Suitable suppository bases include witepsol, macrogol, Tween 61, cocoa butter, lauric acid butter, and glycerol gelatin.

[0020] The dosage of the pharmaceutical composition of the present invention varies depending on the age and sex of the subject to be treated, body weight, the specific disease or pathological condition to be treated, the severity of the disease or pathological condition, the route of administration, and the judgment of the prescriber. Determination of a dosage based on such factors is within the level of skill in the art, and the dosage generally ranges from 0.01 mg / kg / day to approximately 2000 mg / kg / day. A more preferred dosage is from 1 mg / kg / day to 500 mg / kg / day. Administration may be performed once a day or may be divided into several divided doses. The above dosage does not limit the scope of the present invention in any aspect.

[0021] The pharmaceutical composition of the present invention can be administered via various routes to mammals such as mice, livestock, and humans. All modes of administration are contemplated; for example, it can be administered orally, rectally, or via intravenous, intramuscular, subcutaneous, intrauterine dural, or intracerebrovascular injection.

[0022] The present invention provides a health functional food for preventing or improving inflammatory diseases, which comprises the anti-inflammatory composition described above. Further, the anti-inflammatory composition can be added in an amount of 0.001 to 100% by weight to the health functional food of the present invention. The health functional food of the present invention includes forms such as tablets, capsules, pills, and liquids, and examples of foods to which the anti-inflammatory composition of the present invention can be added include various drinks, meat products, sausages, bread, candies, snacks, noodles, ice cream, dairy products, soups, ion drinks, drinking water, alcoholic beverages, chewing gum, tea, vitamin complexes, and the like.

[0023] In another embodiment, the present invention can provide a cosmetic composition comprising the anti-inflammatory composition. The cosmetic composition can be manufactured in any dosage form commonly produced in the art, and can be selected from essences, lotions, emulsions, masks, hand creams, foot creams, lip balms, lipsticks, eyeshadows, eyeliners, eyebrow pencils, blushes, highlighters, general lotions, skin lotions, creams, serums, beauty soaps, softening lotions, medicated lotions, body washes, cleansing foams, cleansing lotions, gels, cleansing oils, cleansing creams, shampoos, conditioners, hair treatments, hair lotions, cleansing tissues, and cleansing waters.

[0024] The cosmetic composition of the present invention may further contain components selected from the group consisting of water-soluble vitamins, oil-soluble vitamins, high molecular weight peptides, high molecular weight polysaccharides, sphingolipids, and seaweed extracts, or conventional cosmetic excipients. [Effects of the Invention]

[0025] The present invention relates to an anti-inflammatory composition containing plum juice as an active ingredient. By further adding Chinese parasol extract, black locust extract, butterbur extract, cinnamon extract, and asparagus extract to the feed composition, it exhibits excellent efficacy in suppressing various inflammatory symptoms while being harmless to the human body. Therefore, it can be used as a therapeutic agent for various inflammatory diseases such as colitis, arthritis, hepatitis, cystitis, nephritis, and dermatitis, as well as a food for improving symptoms, a cosmetic, and the like. [Examples]

[0026] Preferred embodiments of the present invention will be described in detail below. However, the present invention is not limited to the embodiments described herein and may be embodied in other forms. Rather, it is provided to those skilled in the art to fully convey the concept of the present invention so that the content presented herein is thorough and complete.

[0027] <Examples 1-6. Manufacture of anti-inflammatory compositions> Fresh green plums were crushed and juiced, then heated at 90°C for 1 hour. The heated liquid phase was centrifuged at 4°C at 8000 rpm for 30 minutes, filtered through filter paper, and then heated again under the same conditions as the primary heating. After that, the mixture was filtered through a 0.2 μm filter to obtain the plum juice.

[0028] For the Chinese parasol tree, the branches, fruits, and leaves were mixed in equal weights to prepare the sample. For the black locust (acacia), the woody parts of the branches and trunks were prepared, and for cinnamon, the bark of the cinnamon tree was prepared. The Chinese parasol tree, black locust, butterbur (fruit), cinnamon, and asparagus were prepared fresh. Each raw material from the Chinese parasol tree to the cinnamon was heated and extracted at 80°C for 5 hours with 10 kg of water added per 1 kg, and the resulting liquid phase was cooled after removing the residue and used as is.

[0029] A mixture of equal weights of green plums and sugar was left to stand for three months at a cool, shaded room temperature of 25°C. After sufficient juice had been extracted from the plums, the residue was removed and the liquid phase was collected to obtain a plum sugar extract.

[0030] Next, an anti-inflammatory composition was prepared by mixing 100g of the aforementioned plum juice as shown in Table 1 below. [Table 1]

[0031] <Comparative Examples 1-6. Manufacturing of Anti-inflammatory Compositions under Comparative Conditions> A comparative anti-inflammatory composition was prepared under the conditions shown in Table 2. The raw materials and manufacturing methods were the same as in Example 1. [Table 2]

[0032] <Experimental Example 1: Confirmation of Cytotoxicity> 96-well plate (1.5 x 10 3Raw264.7 cells inoculated into cell wells were treated with anti-inflammatory compositions at varying concentrations. After 24 hours of incubation, WST-8 reagent (manufactured by Dojin Chemical Laboratories) was added to the culture medium at 37°C for 1 hour. Absorbance was measured at 450 nm using a microplate reader (BioTek). Subsequently, cell viability was confirmed by measuring absorbance every 24 hours.

[0033] The cell viability for each sample was calculated by converting the values ​​relative to the control group (untreated sample group), where the cell state is set to 100%, and the viability is shown in Table 3 below. [Table 3] Referring to Table 3, it can be confirmed that none of the compositions produced in this invention are cytotoxic.

[0034] <Experimental Example 2: Confirmation of Anti-inflammatory Efficacy> RAW264.7 cells in 2 × 10 6 The cells were dispensed into 60 mm plates at a cells / dish density and allowed to adhere to the plates for 24 hours. To induce an inflammatory response, the cells were treated with a new culture medium containing 1 μg / ml of LPS (lipopolysaccharide) and each anti-inflammatory composition, and incubated for 24 hours.

[0035] The level of NO produced in RAW264.7 cells and present in the culture medium was measured using a Griess reaction-based NO detection kit (Intron 21021). 100 μl of culture medium presumed to contain NO was dispensed into 96-well plates, followed by 50 μl of N1 buffer (sulfanilamide) and reaction at room temperature for 10 minutes. Then, 50 μl of N2 buffer (naphthylethylenediamine) was added and reaction at room temperature for another 10 minutes, after which absorbance was measured at 540 nm. The amount of NO produced was calculated using a standard calibration curve obtained with a nitrite standard. LPS was used as the positive control group.

[0036] [Table 4] Table 4 confirms that the NO-inhibiting activity of the anti-inflammatory compositions of Examples 1-6 is significantly better than that of the compositions of Comparative Examples 1-6.

Claims

1. An anti-inflammatory composition characterized by containing plum juice as an active ingredient, and further containing 20 to 30 parts by weight of Chinese parasol tree extract, 5 to 15 parts by weight of black locust extract, 20 to 30 parts by weight of butterbur extract, 3 to 15 parts by weight of cinnamon extract, and 10 to 20 parts by weight of asparagus extract per 100 parts by weight of plum juice.

2. The anti-inflammatory composition according to claim 1, characterized in that the composition contains a plum sugar extract.

3. A pharmaceutical composition for the prevention or treatment of inflammatory diseases, characterized by containing the composition described in claim 1.

4. The inflammatory disease is selected from the group consisting of atopic dermatitis, edema, dermatitis, allergy, asthma, conjunctivitis, periodontitis, rhinitis, otitis media, pharyngitis, tonsillitis, pneumonia, gastric ulcer, gastritis, Crohn's disease, colitis, hemorrhoids, gout, ankylosing spondylitis, systemic lupus erythematosus, fibromyalgia, psoriatic arthritis, osteoarthritis, rheumatoid arthritis, periarthritis of the shoulder, tendinitis, tenosynovitis, myositis, hepatitis, cystitis, nephritis, Sjögren's syndrome, and multiple sclerosis, as described in claim 3.

5. A health functional food for preventing or improving inflammatory diseases, characterized by containing the composition described in claim 1.

6. The inflammatory disease is selected from the group consisting of atopic dermatitis, edema, dermatitis, allergy, asthma, conjunctivitis, periodontitis, rhinitis, otitis media, pharyngitis, tonsillitis, pneumonia, gastric ulcer, gastritis, Crohn's disease, colitis, hemorrhoids, gout, ankylosing spondylitis, systemic lupus erythematosus, fibromyalgia, psoriatic arthritis, osteoarthritis, rheumatoid arthritis, periarthritis of the shoulder, tendinitis, tenosynovitis, myositis, hepatitis, cystitis, nephritis, Sjögren's syndrome, and multiple sclerosis, as described in claim 5.

7. An anti-inflammatory cosmetic composition characterized by containing the composition described in claim 1.

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