Abstract:
An anti-gout composition including the mixed extract of  Chrysanthemum indicum  and  Cinnamomum cassia  as an active ingredient shows substantially greater inhibitory effects on xanthine oxidase than when  Chrysanthemum indicum  or  Cinnamomum cassia  is used separately. An animal model treated with the mixed extract of  Chrysanthemum indicum  and  Cinnamomum cassia  experienced a decrease in uric acid in serum and in urine. The mixed extract of  Chrysanthemum indicum  and  Cinnamomum cassia  is an active ingredient induced from a natural material, and, accordingly, the mixed extract is safe and a source material therefor is easily provided. Accordingly, the mixed extract can be widely used in gout-related industries.

Description:
TECHNICAL FIELD 
       [0001]    The present invention relates to an anti-gout composition containing a mixed extract of  Chrysanthemum indicum  and  Cinnamomum cassia  as an active ingredient. 
       BACKGROUND ART 
       [0002]    Gout refers to a disease that is caused by deposition of monosodium urate monohydrate crystals (MSU) in articular cartilage, tendons, or surrounding tissues. MSU are generated due to an increase in the uric acid content in blood, wherein the uric acid is the residue of body&#39;s metabolism regarding purine which is intaken through food. The causes of increased gouty arthritis include westernization of diet, increased obesity, aging of the population, increased prevalence of renal disease and hypertension, increased use of diuretics such as thiazide, and low-dose aspirin, etc. 
         [0003]    In addition, gout is a very common inflammatory arthritis among male adults. It is known that the prevalence rate is about 1% to 2% in western countries, and the incidence of disease is lower in Asians compared to Westerners. However, recently, the prevalence rate has been increased in Western and Asian countries due to westernized eating habits and lifestyle. Even in South Korea, 9.3% of adults (14.3% of men, 2.2% of women) are found to have hyperuricemia. Gout is a type of adult disease that easily occurs in obese men. Uric acid increased in the blood is deposited in the form of MSU in the joints or surrounding tissues, causing inflammation, causing pain, swelling, and, in severe cases, deformation of joints. The disease progresses from the period during which the uric acid level in the blood is increased without symptoms, to the period during which acute intermittent arthritis develops and then chronic gouty arthritis develops with deformation of the joints. 
         [0004]    Gout is known to have a clear and successful treatment therefor, but it is often accompanied by other diseases such as hypertension and chronic renal failure. Therefore, side effects of medication should be carefully considered, and, as a non-pharmacological treatment, patients&#39; efforts to change their own lifestyles are essential for good prognosis in long-term treatment. Gout and hyperuricemia show clinical manifestations of hypertension, hyperlipidemia, hyperglycemia, and abdominal obesity. Although they are not the diagnostic criteria for the metabolic syndrome, which is a complex disorder that increases the risk of adult diseases such as arteriosclerotic heart disease and type 2 diabetes, gout and hyperuricemia seem to be closely related to the metabolic syndrome. In South Korea, it was reported that 44% of gout patients were accompanied by metabolic syndrome. Gout usually occurs in the form of acute monoarthritis, but, in some cases, may invade small number of joints or, rarely, multiple joints. Non-steroidal anti-inflammatory drugs (NSAIDs), which are used in the treatment of acute gout, are well known for inhibiting the inflammatory response; colchicine and steroids suppress the inflammatory response by inhibiting the activity and migration of leukocytes, and are all drugs that can effectively treat gout attacks; and selectively, cyclooxygenase (COX-2) inhibitors are known to have the same effect as existing non-steroidal anti-inflammatory drugs. 
         [0005]    In addition, when the uric acid concentration in blood is maintained to a level below the saturation level for a long period of time, the size of existing gout tophi is reduced while acute gouty arthritis is prevented. In the chronic phase of gout, the treatment therefor is carried out to lower the concentration of uric acid in the blood. Uric acid-lowering agents are classified into a xanthine oxidase (XO) inhibitor and a uricosuric agent according to a mechanism. As uric acid-synthesis inhibitors, there are allopurinol, which is widely used, and febuxostat, which has been recently developed as a new drug. Allopurinol is an XO inhibitor that can be effectively used regardless of the cause of hyperuricemia, but the most serious side effect of allopurinol is hypersensitivity syndrome with fever, rash, an increase in eosinophils, hepatitis, renal failure, and the risk of death. Febuxostat is also known as XO inhibitor, but, unlike allopurinol, it is a nonpurine selective blocking agent that is metabolized mainly in the liver to form glucuronide. Most cases of gout progress into the chronic phase, and, even if there are no symptoms, in a prophylactic way, gout is treated by using anti-inflammatory drugs and a method of lowering the uric acid concentration. These prophylactic treatments should be used after the disease has been maintained for a period of time in a calm state, otherwise the gout will recur more severely. However, there is a lot of controversy about the clam period of the illness, and even such prophylactic treatments with existing drugs are not enough to prevent acute episodes of intermittent recurrence of gout, and the technology for inhibiting oxidase, which is a gout-inducing enzyme, with natural products is still insufficient. 
         [0006]    Gout medicines need to be used chronically. Accordingly, side effects caused by cholchine or allopurinol cause serious problems. Therefore, it is necessary to research into and develop natural products that can prevent inflammation for a prolonged period without any side effects. However, the research and the development have not yet been actively performed. As an example of natural products known to date as research results, the extract of  Chrysanthemum indicum  disclosed in Patent No. 2014-0106772 is known to have a gout inhibitory effect. 
         [0007]    Meanwhile,  Cinnamomum cassia  has been known to be effective as an herbal medicine for people with weak constitution and poor blood circulation to promote immunity (Korean Patent No. 0842053). Also, disclosed are a  Cinnamomum cassia  extract-containing composition for prevention and treatment of arteriosclerosis (Korea Patent No. 0294091), a cosmetic composition containing the extract of  Cinnamomum cassia  (Korean Patent Laid-Open No. 2001-0018668), and a composition for promoting oral hygiene containing a nanoparticulate  Cinnamomum cassia  extract (Korea Patent No. 0489267). 
         [0008]    However, as shown according to the present invention, the enhanced effect on gout suppression of the anti-gout composition including the mixed extract of  Chrysanthemum indicum  and  Cinnamomum cassia  as an active ingredient has not been disclosed. 
       DETAILED DESCRIPTION OF THE INVENTION 
     Technical Problem 
       [0009]    The present invention has been made in view of the above-mentioned needs, and an object of the present invention is to provide an anti-gout composition which contains a mixed extract of  Chrysanthemum indicum  and  Cinnamomum cassia  as an active ingredient. More particularly, a xanthine oxidase inhibition rate of the mixed extract of  Chrysanthemum indicum  and  Cinnamomum cassia  was markedly improved as compared with when the extract of  Chrysanthemum indicum  or the extract of  Cinnamomum cassia  is used separately, thereby completing the present invention. 
       Technical Solution 
       [0010]    In order to achieve the above objects, the present invention provides an anti-gout composition which contains a mixed extract of  Chrysanthemum indicum  and  Cinnamomum cassia  as an active ingredient. 
         [0011]    One or more embodiments of the present invention provide a health functional food for prevention or improvement of gout, containing the mixed extract of  Chrysanthemum indicum  and  Cinnamomum cassia  as an active ingredient. 
         [0012]    One or more embodiments of the present invention provide a pharmaceutical composition for prevention or treatment of gout, containing the mixed extract of  Chrysanthemum indicum  and  Cinnamomum cassia  as an active ingredient. 
         [0013]    One or more embodiments of the present invention provide a pharmaceutical composition for prevention or treatment of gout-induced arthritis, containing the mixed extract of  Chrysanthemum indicum  and  Cinnamomum cassia  as an active ingredient. 
       Advantageous Effects 
       [0014]    The present invention relates to an anti-gout composition containing a mixed extract of  Chrysanthemum indicum  and  Cinnamomum cassia . The composition inhibits xanthine oxidase, which is a gout-inducing enzyme, and reduces the amount of uric acid in urine, thereby preventing, improving or treating gout. 
     
    
     
       BRIEF DESCRIPTION OF THE DRAWINGS 
         [0015]      FIG. 1  shows the results obtained by confirming the inhibitory activity of a mixed extract of  Chrysanthemum indicum  ( C. indicum ) and  Cinnamomum cassia  ( C. cassia ) according to the present invention on xanthine oxidase. 
           [0016]      FIG. 2  shows the results obtained by confirming the amount of uric acid contained in the urine according to the treatments of 50, 100, and 200 mg/kg/day of the mixed extract of  Chrysanthemum indicum  and  Cinnamomum cassia  according to the present invention. * and *** indicate that there is a statistically significant difference in the amount of uric acid in the urine between a group treated with allopurinol and a group treated with 50, 100, or 200 mg/kg/day of the mixed extract of  Chrysanthemum indicum  and  Cinnamomum cassia , these groups being compared with a hyperuricemia-induced group; and in the case of *, the p value is less than 0.05, and in the case of ***, the p value is less than 0.001. 
           [0017]      FIG. 3  shows the results obtained by confirming the amount of uric acid contained in the urine according to the treatment of 100 mg/kg/day of the  Chrysanthemum indicum  extract, 100 mg/kg/day of the  Cinnamomum cassia  extract, and 100 mg/kg/day of the mixed extract of  Chrysanthemum indicum  and  Cinnamomum cassia  according to the present invention. * and *** indicate that there is a statistically significant difference in the amount of uric acid in the urine among the allopurinol treatment group, a  Chrysanthemum indicum  extract treatment group, a  Cinnamomum cassia  extract treatment group, and a group treated with various concentrations of the mixed extract of  Chrysanthemum indicum  and  Cinnamomum cassia , these groups being compared with a hyperuricemia-induced group; and in the case of *, the p value is less than 0.05, and in the case of ***, the p value is less than 0.001. 
           [0018]      FIG. 4  shows the results obtained by confirming the amount of uric acid contained in serum according to the treatment of 100 mg/kg/day of the  Chrysanthemum indicum  extract, 100 mg/kg/day of the  Cinnamomum cassia  extract, and 100 mg/kg/day of the mixed extract of  Chrysanthemum indicum  and  Cinnamomum cassia  according to the present invention. * and *** indicate that there is a statistically significant difference in the amount of uric acid in serum among the allopurinol treatment group, a  Chrysanthemum indicum  extract treatment group, a  Cinnamomum cassia  extract treatment group, and a group treated with various concentrations of the mixed extract of  Chrysanthemum indicum  and  Cinnamomum cassia , these groups being compared with a hyperuricemia-induced group; and in the case of *, the p value is less than 0.05, and in the case of ***, the p value is less than 0.001. 
       
    
    
     BEST MODE 
       [0019]    One or more embodiments of the present invention provide an anti-gout composition containing a mixed extract of  Chrysanthemum indicum  and  Cinnamomum cassia  as an active ingredient. 
         [0020]    The mixed extract of  Chrysanthemum indicum  and  Cinnamomum cassia  may be obtained by using water, ethanol, or a mixed solvent thereof, but is not limited thereto. In one embodiment, a mixture weight ratio of  Chrysanthemum indicum  extract and  Cinnamomum cassia  extract in the mixed extract may be in a range of 1:0.1 to 1:10, for example, 1:1. 
         [0021]    The mixed extract of  Chrysanthemum indicum  and  Cinnamomum cassia  according to the present invention may be obtained by obtaining  Chrysanthemum indicum  extract and  Cinnamomum cassia  extract separately and then mixing those extracts. However, in other embodiments,  Chrysanthemum indicum  and  Cinnamomum cassia  may be mixed, and then, the mixture is subjected to an extracting process to obtain the mixed extract of  Chrysanthemum indicum  and  Cinnamomum cassia  according to the present invention. 
         [0022]      Chrysanthemum indicum  used herein refer to every part of the plant including flowers, leaves, stems and roots, and  Cinnamomum cassia  refers to a medicinal material made of a bark of Japanese Cinnamon. 
         [0023]    The mixed extract of  Chrysanthemum indicum  and  Cinnamomum cassia  inhibits xanthine oxidase. 
         [0024]    Another aspect of the present invention provides a health functional food for prevention or improvement of gout, containing the mixed extract of  Chrysanthemum indicum  and  Cinnamomum cassia  as an active ingredient. 
         [0025]    The health functional food for prevention or improvement of gout containing the mixed extract of  Chrysanthemum indicum  and  Cinnamomum cassia  may be prepared in any form selected from beverage, pill, tablet, capsule, and powder, or may be prepared by adding the mixed extract to other food or ingredients of food, and may be suitably prepared according to a conventional method. 
         [0026]    Examples of the foods to which the mixed extract of  Chrysanthemum indicum  and  Cinnamomum cassia  is to be added, include meat, sausage, bread, chocolate, candy, snack, confectionery, pizza, instant noodles, other noodles, gum, dairy products including ice cream, soups, beverages, tea, drinks, alcoholic beverages, and vitamin complexes. The foods used herein refer to any food that is considered as a health-supplementary food in a conventional sense. 
         [0027]    The health functional foods may include various nutrients, vitamins, minerals (electrolytes), synthetic and natural flavors, colorants and enhancers (cheese, chocolate etc.), pectic acid and its salts, alkynic acid and its salts, organic acids, protective colloid thickeners, pH adjusting agents, stabilizers, preservatives, glycerin, alcohols, carbonating agents used in carbonated drinks, and the like. The health functional foods may also include natural fruit juices and fruit flesh for the production of vegetable drinks. These components may be used independently or in combination. 
         [0028]    The health functional food according to the present invention may further contain various flavors or natural carbohydrates as an additional ingredient. Examples of the natural carbohydrates include monosaccharides such as glucose and fructose, disaccharides such as maltose and sucrose, and polysaccharides such as dextrin and cyclodextrin, and sugar alcohols such as xylitol, sorbitol, and erythritol. As a sweetening agent, natural sweetening agents such as thaumatin and stevia extract, or synthetic sweetening agents such as saccharin and aspartame may be used. 
         [0029]    Another aspect of the present invention provides a pharmaceutical composition for prevention or treatment of gout containing the mixed extract of  Chrysanthemum indicum  and  Cinnamomum cassia  as an active ingredient. The pharmaceutical composition according to the present invention may further include a carrier, an excipient, or a diluent in addition to the mixed extract of  Chrysanthemum indicum  and  Cinnamomum cassia . Examples of the carrier, excipient and diluent which may be included in the pharmaceutical composition according to 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, polyvinylpyrrolidone, water, methylhydroxybenzoate, propylhydroxybenzoate, talc, and magnesium stearate. However, the examples of the carrier, excipient and diluent are not limited thereto. 
         [0030]    In addition, the pharmaceutical composition may be administered orally or parenterally in a conventional manner. When the pharmaceutical composition is formulated, a diluent or excipient, such as a filler, an extender, a binder, a wetting agent, a disintegrant, a surfactant, or the like, may be used in the formulation process. Solid formulations for oral administration include tablets, pills, powder, granules, capsules and the like, and these solid formulations are prepared by mixing with at least one excipient selected from starch, calcium carbonate, sucrose, lactose, gelatin, and the like. In addition to simple excipients, lubricants such as magnesium stearate and talc may also be used. Liquid formulations 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. Formulations for parenteral administration include sterile aqueous solutions, non-aqueous solutions, suspensions, emulsions, freeze-dried preparations, and suppositories. Examples of the non-aqueous solvent and the suspending agent include propylene glycol, polyethylene glycol, vegetable oil such as olive oil, injectable ester such as ethyl oleate, and the like. Examples of suppository bases include witepsol, macrogol, tween 61, cacao paper, laurin, glycerogelatin, and the like. 
         [0031]    Another aspect of the present invention provides a pharmaceutical composition for prevention or treatment of gout-induced arthritis containing the mixed extract of  Chrysanthemum indicum  and  Cinnamomum cassia  as an active ingredient. 
         [0032]    Hereinafter, the present invention will be described in more detail with reference to Examples. Those skilled in the art may understand that these examples are only for describing the present invention in detail and that the scope of the present invention is not limited thereto. 
       Example 1. Preparation of Mixed Ethanol Extract of  Chrysanthemum indicum  and  Cinnamomum cassia    
       [0033]    15 l of 70% ethanol was added to 1 kg of  Chrysanthemum indicum  and 1 kg of  Cinnamomum cassia , separately, and each of the mixtures was subjected to an extraction process at a temperature of 85□ for 3 hours. Filtrates obtained therefrom were concentrated under reduced pressure at 50□, thereby obtaining 95 g of  Chrysanthemum indicum  ethanol extract and 77 g of  Cinnamomum cassia  ethanol extract. The  Chrysanthemum indicum  ethanol extract and the  Cinnamomum cassia  ethanol extract were mixed at a weight ratio of 1:1 to prepare a mixed ethanol extract of  Chrysanthemum indicum  and  Cinnamomum cassia.    
       Example 2. Preparation of Mixed Water Extract of  Chrysanthemum indicum  and  Cinnamomum cassia    
       [0034]    15 l of purified water was added to 1 kg of  Chrysanthemum indicum  and 1 kg of  Cinnamomum cassia , separately, and each of the mixtures was subjected to an extraction process at a temperature of 100□ for 3 hours. Filtrates obtained therefrom were concentrated under reduced pressure at 55□, thereby obtaining 146 g of  Chrysanthemum indicum  ethanol extract and 110 g of  Cinnamomum cassia  ethanol extract. The  Chrysanthemum indicum  water extract and the 
         [0035]      Cinnamomum cassia  water extract were mixed at a weight ratio of 1:1 to prepare a mixed water extract of  Chrysanthemum indicum  and  Cinnamomum cassia.    
       Example 3. Inhibitory Activity of Mixed Ethanol Extract of  Chrysanthemum indicum  and  Cinnamomum cassia  on Xanthine Oxidase 
       [0036]    Inhibitory activity of the mixed ethanol extract of  Chrysanthemum indicum  and  Cinnamomum cassia  prepared according to Example 1 on xanthine oxidase was analyzed. The inhibitory activity of xanthine oxidase was analyzed by using the method presented by Sirpe and Della Corte [Stirpe F, Della Corte E. 1969. The regulation of rat liver xanthine oxidase.  J Biol Chem  244: 3855-3563]. 
         [0037]    The mixed extract prepared according to Example 1 was diluted 5-fold and then 0.2 ml of a substrate solution, obtained by dissolving 2 mM of xanthine in 0.1 ml of the diluted solution and 0.6 ml of 0.1 M potassium phosphate buffer (pH 7.5), was added thereto. 0.1 ml of xanthine oxidase (0.2 U/ml) was added thereto and the reaction was carried out at 37□ for 5 minutes. Then, 1 ml of 1N HCl was added to stop the reaction, and the absorbance of uric acid produced in the reaction solution was measured at a wavelength of 292 nm. The inhibitory activity of the mixed extract on xanthine oxidase was shown by the percentage (%) of the absorbance reduction in a group treated with or without a sample. As a control, instead of the mixed extract, the  Chrysanthemum indicum  ethanol extract and the  Cinnamomum cassia  ethanol extract were each diluted in the same manner as applied to the mixed extract, and their inhibitory activities on xanthine oxidase were measured in the same manner as used for the mixed extract. Results obtained therefrom are shown in  FIG. 1 . 
         [0038]    As a result, it was confirmed that, compared with when the same concentration of the  Chrysanthemum indicum  extract or  Cinnamomum cassia  extract was used separately, the mixed extract of  Chrysanthemum indicum  and  Cinnamomum cassia  showed a statistically significant synergistic inhibitory activity on xanthine oxidase. 
       Example 4. Uric Acid-Reducing Effects in Urine of Gout-Induced Animal Model 
       [0039]    Potassium oxonate (150 mg/kg, 0.5% sodium carboxymethylcellulose (CMC-Na with 0.1 M sodium acetate, pH 5)) was intraperitoneally injected to SD-rats, which is a gout animal model, to induce hyperuricemia. After 24 hours, urine was collected from the rats, and hyperacidemia-induced rats were selected therefrom using a QuantiChrom™ Uric Acid Assay Kit (DIUA-250, BioAssay Systems, USA). 200 mg/kg, 100 mg/kg, and 50 mg/kg of mixed extracts of  Chrysanthemum indicum  and  Cinnamomum cassia  (1:1) were administered to the selected experimental animals. Similarly, as a positive control, 50 mg/kg allopurinol was suspended in a 0.01 M PBS buffer containing 0.1% polyoxyethylene sorbitan monolate, and the result was orally administered for 3 days and, on the day just before the autopsy, the rats were fasted for 16 hours. 
         [0040]    After the last oral administration, urine was collected for 2 hours, and the rats were anesthetized with ethyl ether and the blood was taken therefrom. The amount of urine acid in the blood was measured using a QuantiChrom ™ Uric Acid Assay kit (DIUA-250, BioAssay Systems, USA). 
         [0041]    As shown in  FIG. 2 , the amount of uric acid in the urine was increased to about 50 mg/dL by the administration of potassium oxonate, and due to the administration of the mixed extract of  Chrysanthemum indicum  and  Cinnamomum cassia  according to the present invention, the amount of uric acid was decreased to about 32 mg/dL, showing effects similar to those obtained by using allopurinol, which is a positive control. 
       Example 5 Synergistic Effect of  Chrysanthemum indicum  Extract and  Cinnamomum cassia  Extract on Reduction of Uric Acid in Blood and Urine of Gout-Induced Animal Model 
       [0042]    Potassium oxonate (150 mg/kg, 0.5% sodium carboxymethylcellulose (CMC-Na with 0.1 M sodium acetate, pH 5)) was intraperitoneally injected to SD-rats, which is a gout animal model, of gout to induce hyperuricemia. After 24 hours, urine was collected from the rats, and hyperacidemia-induced rats were selected therefrom using a QuantiChrom™ Uric Acid Assay Kit (DIUA-250, BioAssay Systems, USA). 100 mg/kg of the  Chrysanthemum indicum  extract, 100 mg/kg of the  Cinnamomum cassia  extract, and 100 mg/kg of the mixed extract of  Chrysanthemum indicum  and  Cinnamomum cassia  (1:1) were administered to the selected experimental animals. Similarly, as a positive control, 50 mg/kg allopurinol was suspended in a 0.01 M PBS buffer containing 0.1% polyoxyethylene sorbitan monolate, and the result was orally administered for 3 days and, on the day just before the autopsy, the rats were fasted for 16 hours After the last oral administration, urine was collected for 2 hours, and the rats were anesthetized with ethyl ether and the blood was taken therefrom. The amount of urine acid in the blood was measured using a QuantiChrom ™ Uric Acid Assay kit (DIUA-250, BioAssay Systems, USA). 
         [0043]    As shown in  FIG. 3 , the amount of uric acid in the urine was increased to about 53 mg/dL by the administration of potassium oxonate, and due to the administration of the mixed extract of  Chrysanthemum indicum  and  Cinnamomum cassia  according to the present invention, the amount of uric acid was decreased to about 31 mg/dL, showing effects similar to those obtained by using allopurinol, which is a positive control. On the other hand, the  Chrysanthemum indicum  extract and the  Cinnamomum cassia  extract lead to a decrease to 42 mg/dL and 44 mg/dL, respectively. That is, the reduction effect of each of these extracts was smaller than that of the mixed extract of  Chrysanthemum indicum  and  Cinnamomum cassia . Accordingly, it was confirmed that the mixing of the  Chrysanthemum indicum  extract and the  Cinnamomum cassia  extract contributed to the synergistic effect. 
         [0044]    As shown in  FIG. 4 , the amount of uric acid in the blood was increased to about 10.4 mg/dL by the administration of potassium oxonate, and due to the administration of the mixed extract of  Chrysanthemum indicum  and  Cinnamomum cassia  according to the present invention, the amount of uric acid was decreased to about 6.3 mg/dL, showing effects similar to those obtained by using allopurinol, which is a positive control. On the other hand, the  Chrysanthemum indicum  extract and the  Cinnamomum cassia  extract lead to decreases of 8.9 mg/dL and 8.3 mg/dL, respectively. That is, the reduction effect of each of these extracts were smaller than those of the mixed extract of  Chrysanthemum indicum  and  Cinnamomum cassia . Accordingly, it was confirmed that the mixing of the  Chrysanthemum indicum  extract and the  Cinnamomum cassia  extract contributed to the synergistic effect.