201130487 六、發明說明： 【發明所屬之技術領域】 本發明係關於一種黃嗓吟氧化酶抑制劑，其係來自天 然物質’安全性高’並具有優異的黃嘌呤氧化酶抑制活性。 【先前技術】 近年來，飲食生活曰趨豐裕，隨著高卡路里、高蛋白、 高脂肪食品的攝取量增加’高尿酸血症或痛風等因嗓+體 代謝異常所致的疾病也隨之增加。在生物體内，核酸會被 代謝而經嘌呤體、次黃嘌呤、黃嘌呤而變成尿酸，並*** 於尿中。通常’尿酸之產生與***會呈現均衡，血液中的 尿酸值保持於低值，但若尿酸之產生過剩、***降低則 會引起尿酸值異常地變高的高尿酸血症。進而，所蓄積的 尿酸會變成納鹽’並結晶化而沈積於關節内面，如此則會 變成痛風’而發生痛風結節、關節機能障礙、關節變形、 伴隨著劇痛之發作等。 在上述核酸之代謝中’黃嘌呤氧化酶，因為關係到次 黃臂呤至黃嘌呤之變換、及黃嘌呤至尿酸之變換，所以藉 由抑制黃嘌呤氧化酶’而可使尿酸之產生量減少。作為具 有此種黃嘌呤氧化酶抑制作用的藥物，係使用異嘌呤醇。 另一方面，從高尿酸血症或痛風之預防的觀點而言，較期 望是能日常性地攝取而控制尿酸值，所以對於安全性較高 的來自植物的成分也有所探討，例如，已有報告指出，來 3 201130487 自大；匕备、薇ipecz.oscz L.)、及西_洋夏雪草 M Maxim.)的成分具有黃嘌呤氧化酶抑 制活性（專利文獻1及2 )。 [專利文獻1]曰本專利特開2000_291088號公報 [專利文獻2]曰本專利特開2〇〇2_145875號公報 【發明内容】 [發明所欲解決之問題] 本發明係提供一種新穎的黃嘌呤氧化酶抑制劑，其安 全丨生同’忐藉由日常性地持續攝取而有效地抑制黃嘌呤氣 化酶，並有效地預防及治療高尿酸血症及痛風。 [解決問題之技術手段] 本發明者專心進行了研究，結果發現扁實檸檬 知⑽“ Hayata)的料或抽出⑯具有i異的黃以氧化: 抑制作用，進而發現該榨汁或抽出物的黃嘌呤氧化酶抑制 作用係由其所含有的異甜橙素（isosinensetin)、甜橙素 (sinensetln)及檸檬黃素…等多甲氧基黃酮類所發 揮’而完成了本發明。 亦卩本發明係一種黃嘌呤氧化酶抑制劑，其係以扁 實擰檬的榨汁或抽出物作為有效成分。 又，本發明係-種飲食品，其含有上述黃嗓吟氧化酶 抑制劑》 進而本發明得帛高尿酸血症之預防及治療劑、或 201130487 痛風之預防及治療劑，其係黃嘌呤氧化酶抑制劑。 [功效] 本發明之黃嘌呤氧化酶抑制劑，因為安全性高且能藉 由日常性地攝取而有效地使血液中尿酸值降低，所以對於 高尿酸jk症或痛風具有優異的預防及治療效果。 【實施方式】 [實施發明的較佳形態] 本發明的黃嘌呤氧化酶抑制劑，是使用扁實檸檬 (Ciirws c/epresia Hayata)作為原料，並且是以扁實擰檬 的榨汁或抽出物作為有效成分❶扁實檸檬的榨汁，可以是 從果實直接榨汁而得者（一次榨汁），也可以是從榨汁殘渣 再次榨汁者（二次榨汁），但二次榨汁因為黃嘌呤氧化酶抑 制活性高’所以較佳。榨汁的製造，可依照一般方法來進 行’例如可使用螺旋壓榨機、離心壓榨機、帶式壓榨機 (belt-press)等公知的裝置。 另一方面，扁實擰檬的抽出物，是藉由抽出溶劑而將 扁實檸檬之果皮、果粒、梓汁殘液進行抽出而成者。作為 抽出溶劑’以水性溶劑為佳，可例示如：水、乙醇、曱醇 等醇類；丙酮等酮類、乙酸乙酯等酯類、以及此等之混合 液等。此等之中，從作為食品使用的觀點而言，以含水乙 醇、乙醇為佳。抽出操作，通常是使用相對於扁實擰檬為 1〜1 〇倍容量左右的水性溶劑，以溫度為4〜80°C、時間為 201130487 1〜24小時左右進行即可。 作為本發明的扁實擰檬抽出物，可直接使用藉由上述 抽出’谷劑之抽出液，亦可使用進而將該抽出液以合成吸附 樹脂來處理而獲得之吸附區分物（fraction)。此種吸附區分 物因為黃嘌呤氧化酶抑制活性高，所以較佳。合成樹脂吸 附區分物，例如可藉由下述而獲得：將上述抽出液應用於 充填有合成吸附樹脂的管柱之後，視需要而以水等來清洗 該樹煸，接著再以有機溶劑或含水有機溶劑來溶出。 作為上述合成吸附樹脂，可例示如：苯乙烯二乙烯苯 系、甲基丙烯酸酯系等，作為其市售品，在苯乙烯二乙烯 苯系例如可舉出：DIAI〇NHp_2〇 (三菱化學股份有限公司 製造）、DIAI0NHP-21 (三菱化學股份有限公司製造）等， 在曱基丙烯酸酯系則例如可舉出：DIAI〇N Hp2MG (三菱 化子股伤有限公司製造）等。此等之中從適用範圍廣、 可用於食品、醫藥品之分離的觀點而言，又以笨乙烯二乙 烯苯系的合成吸附樹脂為佳，並以使用diai〇n 較 合適。 作為用以從上述合成吸附樹脂中溶出吸附區分物的有 機溶劑或含水有機溶劑，可使用甲醇、乙醇、丙酮、及此 等與水之混合液等。此等之中，卩乙醇、甲醇、或此等與 水之混合液為佳。例如，使用DIAI〇NHp_2〇 (三菱化學股 份有限公司製造）作為合成吸附樹脂時，使用4〇〜的 乙醇水來將吸附成分溶出即可。 作為上述扁實檸檬汁的抽出物的較佳例，可舉出：藉 L 〇 201130487 由濃度為5〜60%的乙醇水來對榨汁殘渣進行抽出之抽出 物、或進而將此抽出物以合成吸附樹脂處理而獲得之吸附 區分物。 上述扁實檸檬的榨汁或抽出物，可直接使用，亦可視 需要而使用蒸餾器或冷凍乾燥機等來濃縮或乾燥、粉末 化。又，視需要而亦可藉由：管柱層析、批次法、液液分 配（liquid-llqu丨d distribution)等一般使用的分離純化手段 來進而純化。 管柱層析，可將合成吸附樹脂、矽膠、(碳十八管 枉）等作為載體’而將該等單獨或組合使用。合成吸附樹脂 方面可例不如.苯乙烯二乙烯苯系曱基丙烯酸酯系等， 作為其市售’在苯乙婦二乙歸苯系例如可舉出：⑴ HP-20 (三菱化學股份有限公司製造）、〇ΐΑ應Ηρ Μ三 菱化學股份有限公司製造m膠方面，市售品可例示 如石夕膠6〇 (默克公司製造）等。〇DS方面，市售品可例示 如佩〇sil-n 5Cl8 HG (和光純藥工業股份有限公司）等。 作職上述載體溶出之移動相，可舉出：甲醇乙醇、 乙酸乙酯、己燒、乙腈等。 如此般藉由管柱層析等分離純化手段，來將上述扁實 檸檬的摊汁或抽出物進行分劃，並將其黃嗓呤氧化酶抑制 活性作為指標而進行純化，藉此獲得含有各種多甲氧基龙 嗣類的區分物。多甲氧基黃嗣類之中，以下述式⑴或(;) 所示之化合物為佳。 W2 ) [化學式1] 201130487 R1BACKGROUND OF THE INVENTION 1. Field of the Invention The present invention relates to a xanthine oxidase inhibitor which is derived from a natural substance which is "safe" and has excellent xanthine oxidase inhibitory activity. [Prior Art] In recent years, dietary life has become more abundant, and with the increase in intake of high-calorie, high-protein, and high-fat foods, diseases such as hyperuricemia or gout have increased due to abnormalities in body metabolism. . In living organisms, nucleic acids are metabolized and become uric acid by carcass, hypoxanthine, and jaundice, and are excreted in the urine. Usually, the production and excretion of uric acid are balanced, and the uric acid value in the blood is kept at a low value. However, if uric acid is excessively produced and excretion is lowered, hyperuricemia in which the uric acid value is abnormally high may be caused. Further, the accumulated uric acid becomes a sodium salt and crystallizes and deposits on the inner surface of the joint, which causes gout, which causes gout nodules, joint dysfunction, joint deformation, and the onset of severe pain. In the metabolism of the above nucleic acid, 'xanthine oxidase, because of the change of the secondary yellow scorpion to the jaundice, and the transformation of jaundice to uric acid, the uric acid production can be reduced by inhibiting xanthine oxidase' . As a drug having such a xanthine oxidase inhibitory action, isodecyl alcohol is used. On the other hand, from the viewpoint of prevention of hyperuricemia or gout, it is more desirable to control the uric acid value on a daily basis, so that a plant-derived component having high safety is also discussed, for example, The report indicates that the components of 3 2011 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 [Patent Document 1] Japanese Laid-Open Patent Publication No. 2000-20988 [Patent Document 2] Japanese Patent Laid-Open Publication No. Hei 2 No. Hei. No. Hei. No. 2-145875 [Draft of the Invention] [Problems to be Solved by the Invention] The present invention provides a novel jaundice An oxidase inhibitor, which is safe and effective, inhibits xanthine gasification enzyme by daily continuous ingestion, and effectively prevents and treats hyperuricemia and gout. [Technical means for solving the problem] The present inventors conducted intensive studies, and as a result, found that the material of the lemon (10) "Hayata" or the extraction of 16 yellow having i is oxidized: inhibition, and then the juice or extract was found. The xanthine oxidase inhibitory effect is achieved by the polymethoxyflavonoids such as isosinensetin, sinensetln, and luciferin contained in the present invention. The present invention relates to a xanthine oxidase inhibitor which is obtained by extracting or extracting a squash lemon. The present invention is a food and drink containing the above-mentioned xanthine oxidase inhibitor. The invention provides a prophylactic and therapeutic agent for hyperuricemia, or a preventive and therapeutic agent for gout of 201130487, which is a xanthine oxidase inhibitor. [Efficacy] The xanthine oxidase inhibitor of the present invention has high safety and can Since the uric acid value in the blood is effectively reduced by daily ingestion, it has an excellent preventive and therapeutic effect on hyperuric acid jk or gout. [Embodiment] [Preferred Embodiment of the Invention] The xanthine oxidase inhibitor of the invention is a juice obtained by using Ciirws c/epresia Hayata as a raw material, and is a juice of a flat lemon or an extract as an active ingredient, and can be Those who directly extract juice from the fruit (one juice) can also be juicer from the juice residue (secondary juice), but the secondary juice is preferred because of the high inhibitory activity of xanthine oxidase. The production of the juice can be carried out in accordance with a general method. For example, a known device such as a screw press, a centrifugal press, or a belt-press can be used. On the other hand, the extract of the flat lemon is The extract of the peel, the fruit, and the clam juice of the flat lemon is extracted by extracting the solvent. The extraction solvent is preferably an aqueous solvent, and examples thereof include alcohols such as water, ethanol, and decyl alcohol; and acetone; An ester such as a ketone or an ethyl acetate, or a mixed liquid thereof, etc. Among these, from the viewpoint of use as a food, it is preferred to use aqueous ethanol or ethanol. The extraction operation is usually carried out using a flat The actual lemon is 1~1 The aqueous solvent having a capacity of about 〇 is carried out at a temperature of 4 to 80 ° C and a time of 201130487 for about 1 to 24 hours. As the flat lemon extract of the present invention, the extract can be used as it is by the above. The extract may be a fraction obtained by treating the extract with a synthetic adsorbent resin. This adsorption fraction is preferred because of its high inhibitory activity of xanthine oxidase. The object can be obtained, for example, by applying the above-mentioned extract liquid to a column packed with a synthetic adsorption resin, and then washing the tree stalk with water or the like as needed, followed by dissolution with an organic solvent or an aqueous organic solvent. Examples of the synthetic adsorption resin include a styrene divinylbenzene system and a methacrylate system. Examples of the styrene divinylbenzene system include DIAI〇NHp_2〇 (Mitsubishi Chemical). (manufactured by Co., Ltd.), DIAI0NHP-21 (manufactured by Mitsubishi Chemical Corporation), etc., in the case of thiol acrylate, for example, DIAI〇N Hp2MG (Mitsubishi Chemicals) Manufactured by the company, Ltd.). Among these, from the viewpoint of wide application range and use for separation of foods and pharmaceuticals, it is preferable to use a synthetic ethylene-diphenylbenzene-based synthetic adsorption resin, and it is preferable to use diii〇n. As the organic solvent or the aqueous organic solvent for eluting the adsorption component from the synthetic adsorption resin, methanol, ethanol, acetone, a mixed liquid with water, or the like can be used. Among these, ethanol, methanol, or a mixture thereof with water is preferred. For example, when DIAI®NHp_2® (manufactured by Mitsubishi Chemical Co., Ltd.) is used as the synthetic adsorption resin, the adsorbed component may be eluted by using 4 〇 of ethanol water. A preferred example of the extract of the above-mentioned flat lemon juice is an extract obtained by extracting the juice residue from ethanol water having a concentration of 5 to 60% by L 〇201130487, or further extracting the extract The adsorption component obtained by the synthesis of the adsorption resin treatment. The juice or extract of the above-mentioned flat lemon can be used as it is, or can be concentrated or dried and pulverized using a distiller or a freeze dryer as needed. Further, it may be further purified by a separation and purification means generally used, such as column chromatography, batch method, or liquid-llqud distribution, as needed. Column chromatography, synthetic adsorption resin, silicone rubber, (carbon 18 tube) or the like can be used as a carrier', and these may be used singly or in combination. Examples of the synthetic adsorbent resin include a styrene divinylbenzene-based fluorenyl acrylate system, etc., and a commercially available 'in the benzene-ethyl bromide-based benzene-based system, for example, (1) HP-20 (Mitsubishi Chemical Co., Ltd.) Manufactured by Μ Η Μ Μ Μ Μ Μ Μ Μ Μ Μ Μ Μ Μ Μ Μ Μ Μ Μ Μ Μ Μ 。 。 。 。 。 。 。 。 。 。 。 。 。 。 In the case of 〇DS, commercially available products can be exemplified, such as 〇 〇 sil-n 5Cl8 HG (Wako Pure Chemical Industries Co., Ltd.). The mobile phase in which the above carrier is eluted may, for example, be methanolic ethanol, ethyl acetate, hexane or acetonitrile. The separation or purification means such as column chromatography is used to separate the juice or extract of the above-mentioned flat lemon, and the xanthine oxidase inhibitory activity is purified as an index, thereby obtaining various kinds. A distinction between polymethoxy steroids. Among the polymethoxyxanthines, a compound represented by the following formula (1) or (;) is preferred. W2) [Chemical Formula 1] 201130487 R1
(1 ) 氳原子或甲氧基） [化學式2] 甲氧基或羥基，R2〜R5獨立表示(1) A halogen atom or a methoxy group) [Chemical Formula 2] A methoxy group or a hydroxy group, and R2 to R5 are independently represented
OCH, (2) 作為上述式（1)的多甲氧基黃酮類，例如可例示：異 甜橙素、甜橙素、[3,铺·5，Μ，·三甲氧基黃_]、[5,6,7,8_ :甲氧基黃刚。又，上述式⑺之化合物為檸檬黃素。 :::上述區分物進而進行純化，而將各化合物單離。此 α物因為分別具有黃嘌呤氧化酶 此等之1插々。， j济性，所以可將 種或2種以上作為本發明的黃嗓 的有效忐八s ^ , 7氧化酶抑制劑 成刀》異甜橙素、甜橙素、檸檬黃素、[3、 一甲氧基黃、及[5,6，Ί四甲氧基相]、a 5’6’4 扁實擰檬的榨汁或抽出物單離或純化而成纟W是從上述 是從扁實檸檬以外的植物抽出而成者 又，亦可以 而成者。此笔夕由 而亦吁以是合成 等之中，又因為黃嘌呤氧化酶抑制、k 而以異甜橙素、甜橙素、檸檬黃素較適合。’性較向’ 異甜撥素、甜橙素、[3，_羥基_5,6,4、三 [5,6,7,8-四甲氡基黃_]，係下述式3〜6所基黃嗣]、 所7^之化合物Ά 8 (3)201130487 [化學式3]OCH, (2) The polymethoxy flavonoids of the above formula (1), for example, may be exemplified by isoamyl orange, sweet orange, [3, sputum, sputum, trimethoxy yellow _], [ 5,6,7,8_ : methoxy Huang Gang. Further, the compound of the above formula (7) is luciferin. ::: The above fractions were further purified, and each compound was isolated. This α substance has a plug of xanthine oxidase, respectively. , j, the species, or more than two species can be used as the effective 忐8s ^, 7 oxidase inhibitor of the scutellariae of the present invention, "iso-sweet orange, sweet orange, taraxanthin, [3, The juice or extract of monomethoxy yellow, and [5,6, decyltetramethoxy phase], a 5'6'4 flat lemon is isolated or purified. Plants other than real lemons can also be grown. This ritual also appeals to synthesis, etc., and because of xanthine oxidase inhibition, k is more suitable for iso-sweet orange, sweet orange, and taraxanthin. 'Sexual orientation' isoflavin, sweet orange, [3,_hydroxy_5,6,4,3[5,6,7,8-tetramethylmethyl]_, is the following formula 3~ 6 base jaundice], 7^ compound Ά 8 (3) 201130487 [Chemical Formula 3]
[化學式6J (4) (S)[Chemical Formula 6J (4) (S)
赞叫的黃嘌呤氧 (6) 將上述扁實#檬的榨二二^ Μ由下述而獲得 橙倉、停權黃素、物或其所含之異對撥素、苗 四P氧基黃刳等多f氧基卷，4，'三甲氧基黃獅J、i5， 法，添加藥理學上可容許的^類，依照公知的製劍學製 劑、網滑劑等而加以製劑 、賦形劑、結合劑、崩解 例如可使用：乳糖、㈣糖時所使用的載體化形劍， 糊精、殿粉、結晶纖維素 201130487 作為結合劑，例如可使用：澱粉、 膠、***膠粉末、 羥基丙基纖維素等；作為崩解劑，例 』那J使用：澱粉、洋 桌、纖維素類等；作為潤滑劑，例如 J1文用.硬脂酸鎂、 滑石、硬脂酸鈣、硬脂酸甘油單酯等。 作為本發明的黃嘌呤氧化酶抑制劑的形態，可作成下 述形態而利用：錠劑、散劑、膠囊、顆粒劑丸劑等固形 劍’·或懸浮劑、乳劑、糖漿等液劑。 使用扁實擰檬抽出物作為有效成分時，相對於全體製 劑，換算為乾燥物以調配2〜80%為佳，更佳為2〇〜4〇%。 又’將上述異甜撥素、甜橙素、檸檬黃素等多甲氧基黃嗣 類作為有效成分時’可單獨調配其中的）種亦可組合調 配2種以上，相對於全體製劑，以調配〇 ι〜5〇%為佳:更 佳為1〜1〇〇/。。 本發明的黃嘌呤氧化酶抑制劑的有效投予量，可根據 患者的年齡、體重、症狀等而適當設定，例如將異甜橙素 作為有效成分時，設為成人每日1300〜 2000mg左右即可。 如此獲得的本發明的黃嘌呤氧化酶抑制劑，可有效地 抑制與在核酸之代謝過程尹從次黃嘌呤變換為黃嘌呤及 從K嘌呤變換為尿酸相關的黃嘌呤氧化酶，而可抑制尿酸 的產生。另一方面，擰檬酸鹽製劑係作為高尿酸血症及痛 風的治療劑而使用於臨床上。溶解於血液中的尿酸是取決 於尿液的pH值，在酸性區域中溶解度低，在中性區域則變 向。檸檬酸鹽製劑是藉檸檬酸循環而被代謝並生成重碳酸 離子，其在體内作為鹼而發揮作用，以將尿液鹼性化，聲、 10 201130487 此促進尿酸的***》扁實擰檬因為含有許多檸檬酸，所以 亦具有使尿液之PH值降低而促進尿酸之***的作用，再配 合上述黃嘌呤氧化酶抑制作用’而可期待具有加倍的高尿 酸血症及痛風的預防及治療效果。 又’藉由直接添加上述黃喝吟氧化酶抑制劑或再加 入各種營養成分而添加至公知的飲食品，而可作為對高尿 酸血症及痛風之預防及治療具有作用的飲食品。作為飲食 品的較佳例’可例示：$汁飲料、清涼飲料、濃縮飲料、 營養飲料、酒精飲料、σ香糖、糖果等。 [實施例] 以下舉出實施例以更詳細地說明本發明，但本發明並 不受此等實施例之任何限定。 實施例1 扁實檸檬榨汁的製造： ^將扁實檸檬的果實以帶式壓榨機加以榨汁’而獲得一 果汁。進而，將該榨汁湩滓以螺旋壓榨機（SRE_〗50S， °工程（Shinwa Engineering)股份有限公司）再次壓榨， 獲得一次榨汁。對於該二次榨汁進行檸檬酸含量之測 1 ay , A，…果為1.66%(w/v)。擰檬酸含量是藉由酸滴定來測 疋酸度，再換算為檸檬酸而求得（檸檬酸換算係數為 0-0064) 〇 201130487 試驗 例 1 黃嘌呤氧化酶抑制活性的測定： 對於實施例1所獲得之扁實檸檬的一次果汁及二次榨 汁’變更黃嘌呤氧化酶分析套組（Xanthine 0xidase Assay Kit) ( Funakoshi Co.，Ltd，）而測定黃嘌呤氧化酶抑制活性。 亦即，混合50 #】之試料與5〇私1之藉由x〇樣品緩衝液（χ〇 Sample Buffer)製備的黃嘌呤氧化酶（〇 1 於其 t添加50#1之藉由X0分析緩衝液（χ〇 Assay Buffer·)、檢 測劑（Detector)及辣根過氧化酶（H〇rseradish per〇xidase)所 製備的反應試液’於37°C使其反應45分鐘，再藉螢光分 光光度計，以激發波長540 nm、螢光波長590 nm來測定。 對照組係使用XO樣品緩衝液來取代試料溶液，並且作為 各空白組（blank)而添加χ〇樣品緩衝液以取代酵素溶液， 並進行同樣的操作。如上述般進行測定並藉由下述式來求 取抑制率，結果平均每从g/ ml之果汁中，一次果汁 為 41.4°/。、二次榨汁為 65 8〇/〇。 (黃嘌呤氧化酶抑制率） 抑制率（。/〇 = _ (C_D)] /(A—Β)Χ 1〇〇 其中’ A ’對照溶液在590 nm中之螢光度、β :對照 溶液空白組在590 nm中之螢光度、c:試料溶液在59〇nm 中之螢光度、D:試料溶液空白組在59〇 nm中之螢光度。 實施例 2 扁實檸檬抽出物的製造（1): 〃 12 201130487 於277 kg之扁實檸檬榨汁殘渣中添加56〇l之5〇%乙 醇，靜置過夜（overnight)再進行抽出。藉由過濾器進行過 濾後，將藉由超高速離心而澄清者作為抽出液。將抽出液 供應至DIAION HP20管枉（15L)，而獲得未吸附之通透 液、與95〇/〇乙醇溶出液（7〇L)。將此95%乙醇溶出液藉蒸 發器（EVAPOR)濃縮至15L而獲得吸附區分物。 實施例 3 扁實檸檬抽出物的製造（2): 將8 g之實施例2所獲得之吸附區分物，溶解於水與 乙醇之混合溶劑’再使其通入至DIAI〇N HP20層析管柱 (5.9x25 cm )。接著，階段性地通入：2〇%、4〇%、6〇%、 80%、95%之乙醇、丙酮，將管柱通透液每1〇〇〇 進行分 取，而分劃為6份區分物，將各區分物從最初的通透液起 依序編號（Fr. 1〜6)。對於各區分物，與試驗例}同樣地 測定黃嘌呤氧化酶抑制活性》結果顯示於第丨圖。將抑制 活性最強的Fr. 4 ( 932 mg)以矽膠層析管柱（4.8x17 cm， 溶劑系為己烷/乙酸乙酯、及乙酸乙酯/甲醇）來分劃為 8份區分物（Fr· 4-1〜4-8)，而獲得抑制活性強的Fr, 4-7 (383 mg)。將Fr. 4-7再次以矽膠層析管柱（2.6x15 cm , 溶劑系為已酸乙醋/甲醇）來分劃為7份區分物（Fr. 4-7-1 〜4-7-7 )’而獲得抑制活性強的Fr. 4-7-2 ( 66 mg)。進而 將 Fr. 4-7-2 用 Wakosil-Π 5C18 HG( 10x250 mm)藉由 HPLC(高效能液相色層分析）來分劃^ 13 201130487 4-7-2-4) ’結果獲得具有抑制活性的Fr. 4-7-2-3 ( 6 mg)。 對於此區分物，藉由NMR(核磁共振光譜）及MS(質譜分析） 之分析來鑑別異甜撥素❶同樣地從Fr. 5、Fr. 1及Fr. 4分 劃純化’而分離出甜橙素、檸檬黃素及[3，_羥基_5 6 4、三 甲氧基黃酮]«異甜橙素、甜橙素、檸檬黃素之IC5〇 (5〇% 抑制濃度）顯示於下述表1。 1] _ 一 ICS0("g/ml) ___ 異甜橙素 15.8 _ 甜橙素 22.4 檸檬黃素 24.4 配 方 例 1 含扁實檸檬二次榨汁之飲料的製備： 製備下述配方之飲料。 (每 100 ml) 5.0 g 4.0 g 0.05 g 0.1 g 100g (以純水調整至l〇0g) (配方） (1) 實施例1之扁實檸檬二次榨汁 (2) 果糖 (3) L-抗壞血酸 (4) 香料 (5) 純水 合計 [產業上之可利用性] 本發明的黃嘌呤氧化酶抑制劑，其安全性高，能藉由 曰常性地攝取而控制血液中的尿酸值，所以作為用於預防 及治療高尿酸血症或痛風的醫藥或食品而具有作用。 201130487 【圖式簡單說明】 第1圖是顯示在實施例2中將扁實擰檬抽出液的合成 樹脂吸附區分物，以管柱層析來分劃而得之區分物的黃嘌 呤氧化酶抑制活性的圖。 【主要元件符號說明】 無 15The singular scutellariae (6) obtained the above-mentioned sturdy stalks of the stalks of the stalks of the stalks of the stalks of the stalks of the stalks of the stalks A variety of f-oxygen rolls such as Astragalus membranaceus, 4, 'trimethoxy yellow lion J, i5, method, adding pharmacologically acceptable class, according to the well-known sword preparation, net slip agent, etc. For the preparation of the agent, the binder, and the disintegration, for example, a carrier-shaped sword used for lactose or (iv) sugar, dextrin, temple powder, and crystalline cellulose 201130487 can be used as a binding agent, for example, starch, gum, gum arabic powder can be used. , hydroxypropyl cellulose, etc.; as a disintegrating agent, for example, J: starch, ocean table, cellulose, etc.; as a lubricant, for example, J1 text. Magnesium stearate, talc, calcium stearate, Stearic acid monoglyceride and the like. The form of the xanthine oxidase inhibitor of the present invention can be used as a solid agent such as a tablet, a powder, a capsule or a granule pellet, or a liquid preparation such as a suspension, an emulsion or a syrup. When the flat lemon extract is used as the active ingredient, it is preferably 2 to 80%, more preferably 2 to 4% by weight, based on the total amount of the whole system. In addition, when the polymethoxyxanthine such as the isodatin, the sucrose, or the taraxanthin is used as the active ingredient, the above-mentioned types may be blended in combination of two or more kinds, and the whole preparation may be used. It is better to mix 〇ι~5〇%: more preferably 1~1〇〇/. . The effective administration amount of the xanthine oxidase inhibitor of the present invention can be appropriately set according to the age, body weight, symptoms, and the like of the patient. For example, when iso-sucrose orange is used as an active ingredient, it is set to be about 1300 to 2000 mg per day for an adult. can. The xanthine oxidase inhibitor of the present invention thus obtained is effective for inhibiting xanthine oxidase associated with the conversion of Yin from hypoxanthine to xanthine and from K嘌呤 to uric acid in the metabolic process of nucleic acid, and inhibiting uric acid The production. On the other hand, the citrate preparation is used clinically as a therapeutic agent for hyperuricemia and gout. The uric acid dissolved in the blood depends on the pH of the urine, and the solubility in the acidic region is low, and it is changed in the neutral region. The citrate preparation is metabolized by the citric acid cycle to form bicarbonate ions, which acts as a base in the body to alkalinize the urine, sound, 10 201130487, which promotes the excretion of uric acid. Because it contains a lot of citric acid, it also has the effect of lowering the pH of urine and promoting the excretion of uric acid. Together with the inhibition of xanthine oxidase, it can be expected to prevent and treat hyperuricemia and gout. effect. In addition, by adding the above-mentioned yellow sputum oxidase inhibitor or adding various nutrients to a known food or drink, it can be used as a food or drink for the prevention and treatment of hyperuricemia and gout. Preferred examples of the food and drink are exemplified by a juice drink, a refreshing drink, a concentrated drink, a nutritional drink, an alcoholic beverage, a swig, a candy, and the like. [Examples] Hereinafter, the present invention will be described in more detail by way of examples, but the present invention is not limited by the examples. Example 1 Manufacture of flat lemon juice: ^ Juice the fruit of a flat lemon with a belt press to obtain a juice. Further, the juice extract was again pressed by a screw press (SRE_) 50S, ° (Shinwa Engineering Co., Ltd.) to obtain a single juice. For the secondary juice, the citric acid content was measured 1 ay , A, ... the fruit was 1.66% (w / v). The citric acid content is determined by acid titration to determine the acidity of the ruthenium and then converted to citric acid (citric acid conversion factor is 0-0064) 〇201130487 Test Example 1 Determination of xanthine oxidase inhibitory activity: For Example 1 The xanthine oxidase inhibitory activity was measured by the obtained single juice and secondary juice 'Xanthine 0xidase Assay Kit' (Funakoshi Co., Ltd.). That is, the mixed 50#] sample and the 5〇 private 1 xanthine oxidase prepared by x〇 sample buffer (χ〇Sample Buffer) (〇1 is added to 50#1 by X0 analysis buffer The reaction solution prepared by the liquid (χ〇Assay Buffer·), the detector (Detector) and the horseradish peroxidase (H〇rseradish per〇xidase) was reacted at 37 ° C for 45 minutes, and then the spectrophotometry was carried out by fluorescence. The measurement was performed at an excitation wavelength of 540 nm and a fluorescence wavelength of 590 nm. The control group used XO sample buffer to replace the sample solution, and as a blank, a ruthenium sample buffer was added to replace the enzyme solution, and The same operation was carried out. The measurement was carried out as described above, and the inhibition rate was determined by the following formula. As a result, the average juice per juice was 41.4°/g from the g/ml juice, and the secondary juice was 65 8〇/嘌呤. (Huangqi oxidase inhibition rate) Inhibition rate (./〇= _ (C_D)] /(A—Β)Χ 1〇〇 The fluoresce of the 'A' control solution at 590 nm, β: control solution Fluorescence of blank group at 590 nm, c: luminosity of sample solution at 59 〇 nm, D: sample dissolution Fluorescence of the blank group at 59 〇 nm. Example 2 Manufacture of flat lemon extract (1): 〃 12 201130487 Add 56 〇l of 5 〇% ethanol to 277 kg of flat lemon juice residue, static The mixture was taken overnight and then extracted. After filtration by a filter, the clarified by ultracentrifugation was used as the extract. The extract was supplied to DIAION HP20 tube (15 L) to obtain unadsorbed permeability. Liquid, and 95 〇 / 〇 ethanol eluate (7 〇 L). This 95% ethanol eluate was concentrated to 15 L by means of an evaporator (EVAPOR) to obtain an adsorption fraction. Example 3 Production of a flat lemon extract (2 ): 8 g of the adsorption fraction obtained in Example 2 was dissolved in a mixed solvent of water and ethanol' and then passed to a DIAI〇N HP20 chromatography column (5.9 x 25 cm). Then, stepwise Access: 2〇%, 4〇%, 6〇%, 80%, 95% ethanol, acetone, the column permeate is divided every 1〇〇〇, and the division is 6 parts, Each of the different components was numbered sequentially from the first permeation liquid (Fr. 1 to 6). For each of the classifications, the jaundice was measured in the same manner as in the test example}. The results of oxidase inhibitory activity are shown in the figure. The most potent inhibitory activity of Fr. 4 (932 mg) is a ruthenium chromatography column (4.8 x 17 cm, solvent hexane/ethyl acetate, and ethyl acetate/ Methanol) was divided into 8 parts (Fr· 4-1 to 4-8), and Fr, 4-7 (383 mg) having strong inhibitory activity was obtained. Fr. 4-7 was again divided into 7 parts by gel column (2.6 x 15 cm, solvent ethyl acetate / methanol) (Fr. 4-7-1 ~ 4-7-7 '' and obtain Fr. 4-7-2 (66 mg) with strong inhibitory activity. Further, Fr. 4-7-2 was divided by Wakosil-Π 5C18 HG (10×250 mm) by HPLC (High Performance Liquid Chromatography) ^ 13 201130487 4-7-2-4) 'Results obtained with inhibition Active Fr. 4-7-2-3 (6 mg). For this distinction, the analysis of NMR (nuclear magnetic resonance spectroscopy) and MS (mass spectrometry) was used to identify isodextrin quinones, which were similarly purified from Fra. 5, Fr. 1 and Fr. 4 to separate sweets. The orange, luciferin, and [3,_hydroxy_5 4 4 , trimethoxy flavonoids] «isodansine, sucrose, and luciferin IC5 〇 (5 〇% inhibitory concentration) are shown in Table 1 below. 1] _ ICS0("g/ml) ___ Iso-orange orange 15.8 _ Sweet orange 22.4 Lutein 24.4 Formulation Example 1 Preparation of a beverage containing a double lemon juice: Prepare a beverage of the following formula. (per 100 ml) 5.0 g 4.0 g 0.05 g 0.1 g 100 g (adjusted to 100 g with pure water) (recipe) (1) The second lemon juice of Example 1 (2) Fructose (3) L- Ascorbic acid (4) Perfume (5) Pure water total [Industrial Applicability] The xanthine oxidase inhibitor of the present invention has high safety and can control the uric acid value in the blood by constantly ingesting it. Therefore, it has a role as a medicine or food for preventing and treating hyperuricemia or gout. 201130487 [Simplified description of the drawings] Fig. 1 is a diagram showing the inhibition of xanthine oxidase of a differentiated synthetic resin adsorbed material in the second embodiment, which is classified by column chromatography. The map of activity. [Main component symbol description] None 15