九、發明說明： 【發明所屬之技術領域】 本發明係關於一系列白鞭珊瑚酯及其分離方法，其特別是關 於該白鞭珊瑚酯為白鞭珊瑚酯c及白鞭珊瑚酯D，及自白鞭珊瑚 提取該白鞭珊瑚酯C及白鞭珊瑚酯D之分離方法。 【先前技術】 一般而言，萜烯^φεηοίί!〕及其衍生物〔derivatives〕係廣 泛存在於植物體内的天然來源碳氫化合物，其可自許多植物提取 獲得；特別是’其可自針葉樹〔coniferous tree〕取得。事實上， 一般樹脂和由樹脂產生的松節油的主要成分為萜烯。在應用上， 各類萜類化合物具有重要的生理活性，其為研究天然產物及開發 新藥的重要來源。 近年的研究顯示’除了在植物中大量存在萜類化合物外，在 海洋生物體内中亦大量存在萜類化合物。因此，自海洋生物體可 適當提取出大量的萜類化合物。目前的研究統計數據顯示，已知 的萜類化合物的總數可能超過22000種。 舉例而言’萜稀衍生物〔tei*penoid derivative〕可提取自台灣 產八放珊瑚〔Formosan octocoral〕’其包含自紅鞭珊湖或白鞭珊 糊提取薛類化合物。然而’就自紅鞭珊湖或白鞭珊湖提取祐類化 合物而言’必須以適當的技術手段方能成功取得，以期不破壞原 化合物結構及生物活性〔biological activity〕，並仍能保有天然抗 氧化效力〔inhibitory effect〕。 目前為止’有關自紅鞭珊瑚或白鞭珊蝴提取祐類化合物技術 極少揭示於世界各國專利之技術内容；例如：中國專利第 1353.355 cm9483〇2號、第CN1715290號及第CNm5281號等。因此，董 於自紅鞭珊喊白鞭珊喊取新式㈣化合物，或對於開發新^ 提取技術方法自紅鞭珊喊自鞭响提取㈣化合物 = 強烈的需求。 ^存在 有鏗於此’本發明為了考量上述需求，其提供-系列白鞭珊 糊S曰化合物’該白鞭珊触為白鞭珊躺C及白鞭珊㈣D，並 提供自白鞭_提取該白鞭珊_ c及白鞭珊_ d之分離方法Nine, the invention relates to: [Technical field of the invention] The present invention relates to a series of white whip coral esters and a method for separating the same, in particular, the white whip coral ester is white whip coral ester c and white whip coral ester D, and The separation method of the white whip coral ester C and the white whip coral ester D was extracted from the white whip coral. [Prior Art] In general, terpenes ^φεηοίί!] and derivatives thereof are naturally derived hydrocarbons widely present in plants, which can be extracted from many plants; in particular, they can be derived from conifers. [coniferous tree] obtained. In fact, the main component of the general resin and turpentine produced from the resin is terpene. In application, various terpenoids have important physiological activities, which are important sources for the study of natural products and the development of new drugs. Recent studies have shown that in addition to the presence of terpenoids in plants, a large number of terpenoids are present in marine organisms. Therefore, a large amount of terpenoids can be appropriately extracted from marine organisms. Current research statistics show that the total number of known terpenoids may exceed 22,000. For example, a 'tei*penoid derivative' can be extracted from Taiwan's Formosan octocoral, which contains a Xue compound extracted from the red whipping lake or the white whip. However, 'in terms of extracting the compound from the red whip lake or the white whip lake, ' must be successfully obtained with appropriate technical means, so as not to destroy the original compound structure and biological activity, and still retain the natural Inhibitory effect. So far, the technology for extracting the compound from the red whip coral or the white whip butterfly has rarely revealed the technical content of patents in countries around the world; for example, Chinese patents No. 1353.355 cm9483〇2, CN1715290 and CNm5281. Therefore, Dong Yuhong whip Shan shouted white whip Shan shouted to take a new formula (IV) compound, or for the development of new ^ extraction technology method from the red whip shouting from the whiplash extraction (four) compound = strong demand. ^There is a reason for this. In order to consider the above requirements, the present invention provides a series of white whip smear compounds. The white whip is touched as a white whip and a white whip (four) D, and provides a self-white whip. White whip _ c and white whip _ d separation method
’如此本發明能達成提供新式白鞭珊_旨及提供分離該白鞭珊 酯技術方法之目的。 【發明内容】 本發明之主要目的係提供—種白鞭珊咖旨系列，該白鞭珊蝴 龜為白鞭__ c及白鞭珊_ D，其達成提供新式白鞭珊_ 之目的。 本發明之另__目的係提供—種白鞭珊咖旨分離方法，其自白Thus, the present invention achieves the object of providing a novel white whip and providing a method for separating the white whip ester. SUMMARY OF THE INVENTION The main object of the present invention is to provide a series of white whip-chantles, which are white whip __ c and white whip _ D, which achieve the purpose of providing a new type of white whip. Another object of the present invention is to provide a method for separating a white whip and a confession
鞭喃提取該白鞭珊_旨C及自伽_ D，其達成提供錄白 鞭珊瑚酯技術方法之目的。 為了達成上述目的’本發明之白鞭珊咖旨為白鞭珊躺C及 白鞭珊_ D，且該白鞭珊_ C及白鞭珊躺D提取自白鞭珊 為了達成上述目的’本發明之白鞭珊躺分離方法包含 如下： 自白鞭珊蝴進行初步萃取’以提取-初步萃取品； 將該初步萃取品進行分離’以選取__分離品； 將該分離品進行分離，以取得數個第一分部提取液； 6 將適當選擇該第一分部提取液之一再進行分離，並進行提取 ，以取得白鞭珊瑚酯。 另外，將適當選擇該第一分部提取液之一再進行分離，以取 得數個第二分部提取液； 將適當選擇該第二分部提取液之一進行純化，以取得另一白 鞭珊瑚酯。 本發明較佳實施例之白鞭珊瑚選自乾白鞭珊瑚碎片。 本發明較佳實施例係將該白鞭珊砌以甲醇及二氯子烷之混合 液進行萃取，以取得該初步萃取品。 本發明較佳實施例係將該初步萃取品於乙酸乙酯〔Et0AC〕 及水中進行分離，以選取一 EtOAc分離品。 本發明較佳實施例係將該EtOAc分離品於矽膠凝體進行分離 ，並以正己烷-乙酸乙酯進行洗脫，以取得數個該第一分部提取液 〇 本發明較佳實施例係將該第二分部提取液以二氣甲烷及乙酸 乙酯之混合液進行純化。 【實施方式】 為了充分瞭解本發明’於下文將例舉較佳實施顺配合所附 圖式作詳細說明，且其並非用以限定本發明。 本發明之白鞭珊构6旨系列包含白鞭珊瑚醋c及白鞭珊蝴醋D :本發明於此定義、、包含々為自白鞭珊射提取兩種白鞭珊_旨 。是以，該白鞭珊蝴醋為可自白鞭珊期可提取白鞭珊蝴酉旨c ;或， 該白鞭珊瑚酯為可自白鞭珊瑚可提取白鞭珊瑚酯D。本發明以適 當方式提取該白鞭珊瑚酯，且將該白鞭珊瑚酯製成適當製品，其 不破壞原化合物結構及生物活性，並仍能保有天然抗氧化效力。 本發明第一較佳實施例之白鞭珊瑚酯為白鞭珊瑚酯C，而本 發明第二較佳實施例之白鞭珊瑚酯為白鞭珊瑚酯D。本發明第一 較佳實施例及第二較佳實施例之白鞭珊瑚酯C及白鞭珊瑚酯d係 利用本發明分離方法或其他等效方法提取自白鞭珊瑚。該白鞭珊 构5曰C及白鞭珊蝴醋〇可利用光譜分析〔Spectrai data analysis〕 方式確認其各自化學結構。 本發明較佳實施例之白鞭珊蝴•沿〕分佈於 〇灣海及西太平洋沿海區域。本發明較佳實施例之白鞭珊瑚選 自乾白鞭珊瑚碎片〔freeze-dried «/imcee/Za 〕，但其並非用 以限制本發明。 本發明定義白鞭珊瑚酯C〔fragilide C〕具有化合物之分子 气’並以該分子式表示該白鞭珊瑚酯匸。本發明定義 白鞭珊瑚酯c僅只區別白鞭珊瑚酯之種類，其並非用以表示該白 鞭珊瑚酯之種類特性。 凊參照第1圖所示，第1圖揭示本發明第一較佳實施例之白 鞭_8旨C之化學式之結構示意圖。 本發明定義白鞭珊瑚酯D〔fragilide D〕具有化合物之分子 ' qitinClO,2’並以該分子式表示該白鞭珊瑚酯d。本發明定 義白鞭珊瑚§旨D僅只區別白鞭珊賴之種類，其並翻以表示該 白鞭珊蝴酯之種類特性。 凊參照第2圖所示，第2圖揭示本發明第一較佳實施例之白 鞭珊之化學式之結構示意圖。 舉例而言，本發明較佳實施例之白鞭珊蝴酯之分離方法首先 1353355 於室溫將乾白鞭珊瑚碎片以甲醇及二氣甲烷〔l : l〕之混合液進 行萃取，以取得一初步萃取品〔preliminary residue〕；將該初步萃 取品於乙酸乙酯〔EtOAc〕及水中進行分離，以選取一 Et〇Ac分 離品〔EtOAc residue〕；將該EtOAc分離品於石夕膠凝體〔suica㈣ 〕進行分離，並以正己烷-乙酸乙酯〔50 : 1〕進行洗脫，以取得數 個第一分部提取液〔first fraction〕，其包含π個第一分部提取液 〔Fr. A至Fr.Q〕；其中將該第一分部提取液之Fr.K進行分離， 並以二氣曱烷-乙酸乙酯〔15 : 1〕進行洗脫，以取得白鞭珊瑚酯D 〇 另外，將該第一分部提取液之Fr· K再於矽膠凝體進行分離 ，並以一乳甲烧及乙酸乙醋〔20 : 1〕進行洗脫，以取得數個第二 分部提取液〔second fraction〕，其包含23個第二分部提取液〔Fr K1至Fr. K23〕；將該第二分部提取液之Fr· K7以正相高效能液相 層析系統〔NP-HPLC〕進行純化，並利用二氣甲烷及乙酸乙酯混 合液k供本發明第二較佳實施例之白鞭珊蝴醋C。 前述兩種較佳實施例之分離方法可適當提取本發明第—較佳 實施例及第二較佳實施例之白鞭珊瑚酯C及白鞭珊瑚酯D。 表1 .本發明較佳實施例白鞭珊蝴酯C之生物活性〔 biological activity〕表現於抗氧化作用〔inhibit〇ry他⑴之量測 結果。 化合物 抑制超氧陰離子產生之百分比(O/〇)a 白鞭珊瑚酯C -__H.61+2.80c a .於濃度_g/mL之抑制氧化百分比〔恤％〕，其誤差範 mean+SEM ( n = 3 ] 0 9 1353355 .. •, 表1顯示本發明較佳實施例白鞭珊瑚酯之生物活性於濃度 10pg/mL實驗中檢測人類嗜中性白血球〔human neutrophils〕之超 軋陰離子產生〔SUperoxide anion generation〕對於 fMet-Leu-Phe/Cytochalastin B所形成的抑制超氧陰離子產生之百 分比；其中白鞭珊瑚酯C具有11.61 士2.80%。 上述實驗數據為在特定條件之下所獲得的初步實驗結果，其 僅用以易於瞭解或參考本發明之技術内容而已，其尚需進行其他 .實驗。該實驗數據及其結果並非用以限制本發明之權利範圍。 ® 前述較佳實施例僅舉例說明本發明及其技術特徵，該實施例 之技術仍可適當進行各種實質等效修飾及/或替換方式予以實施； 因此’本發明之權利範圍須視後附申請專利範圍所界定之範圍為 準。 10 1353355 【圖式簡單說明】 第1圖：本發明第一較佳實施例之白鞭珊瑚酯c之化學式之 結構示意圖。 第2圖：本發明第二較佳實施例之白鞭珊瑚酯D之化學式之 結構示意圖。 【主要元件符號說明】The whip is extracted from the white whip _ _ C and self gamma _ D, which achieves the purpose of providing the technical method of recording whit coral ester. In order to achieve the above object, the white whip of the present invention is intended to be a white whip and a white whip _ D, and the white whip _ C and the white whip lie D are extracted from the white whip to achieve the above purpose. The method for separating the white whip is as follows: a preliminary extraction from the white whip butterfly to extract the preliminary extract; separating the preliminary extract to select the __separate; separating the fraction to obtain the number The first fraction extract; 6 one of the first fraction extracts is appropriately selected and separated, and extracted to obtain the white whip coral ester. In addition, one of the first fraction extracts is appropriately selected and separated to obtain a plurality of second fraction extracts; one of the second fraction extracts is appropriately selected for purification to obtain another white whip coral ester. The white whip coral of the preferred embodiment of the invention is selected from the group consisting of dry white whip coral fragments. In a preferred embodiment of the invention, the white whip is extracted with a mixture of methanol and dichlorocyclohexane to obtain the preliminary extract. In a preferred embodiment of the invention, the preliminary extract is separated in ethyl acetate [Et0AC] and water to give an EtOAc isolate. In a preferred embodiment of the invention, the EtOAc fraction is separated in a ruthenium gel and eluted with n-hexane-ethyl acetate to obtain a plurality of the first fraction extracts. The second fraction extract was purified as a mixture of dimethane and ethyl acetate. The invention is described in detail in the following description of the preferred embodiments of the invention, and is not intended to limit the invention. The white whip stalk 6 series of the present invention comprises white whip coral vinegar c and white whip berry vinegar D: the invention is defined herein, and comprises two kinds of white whip stalks. Therefore, the white whip berry vinegar can be extracted from the whitish whipping period, or the white whip coral ester can be extracted from the white whip coral. The present invention extracts the whitish coral ester in an appropriate manner, and the whispering coral ester is made into a suitable product which does not destroy the structure and biological activity of the original compound, and still retains the natural antioxidant effect. The white whip coral ester of the first preferred embodiment of the present invention is a white whip coral ester C, and the white whip coral ester of the second preferred embodiment of the present invention is a white whip coral ester D. The first preferred embodiment and the second preferred embodiment of the present invention are the white whip coral C and the white whip coral ester d. The white whip coral is extracted by the separation method of the present invention or other equivalent method. The white whip and the white whip vinegar can be confirmed by spectra analysis (Spectrai data analysis). The preferred embodiment of the present invention is distributed in the coastal area of the Tsuen Wan Sea and the western Pacific Ocean. The white whip coral of the preferred embodiment of the present invention is selected from freeze-dried «/imcee/Za, but it is not intended to limit the invention. The present invention defines that the white crater C (fragilide C) has a molecular gas of the compound ' and the whisperic acid oxime is represented by the formula. The present invention defines that the white whip coral ester c only distinguishes the species of the white whip coral ester, and is not intended to indicate the species characteristics of the white whip coral ester. Referring to Fig. 1, there is shown a schematic view showing the structure of the chemical formula of the white whip _8 of the first preferred embodiment of the present invention. The present invention defines that the whigilide D has the molecule 'qitinClO, 2' of the compound and represents the whitish coral ester d in this formula. The present invention defines that the white whip coral § D only distinguishes the species of the white whip, and it is turned over to indicate the species characteristics of the white whip. Referring to Fig. 2, Fig. 2 is a schematic view showing the structure of the chemical formula of the white whip in the first preferred embodiment of the present invention. For example, the method for separating the white whip butterfly ester of the preferred embodiment of the present invention firstly extracts the dry white whip coral pieces with a mixture of methanol and diqi methane [l: l] at room temperature to obtain a a preliminary extract (preliminary residue); the preliminary extract is separated in ethyl acetate [EtOAc] and water to obtain an Et 〇Ac isolate (EtOAc residue); Suica (4)] was separated and eluted with n-hexane-ethyl acetate [50:1] to obtain several first fraction extracts containing π first fraction extracts [Fr. A to Fr. Q]; wherein the Fr. K of the first fraction extract is separated and eluted with dioxane-ethyl acetate [15:1] to obtain a white whip coral ester D 〇 In addition, the Fr·K of the first fraction extract is separated from the ruthenium gel, and eluted with a milk emulsion and ethyl acetate [20:1] to obtain a plurality of second fraction extracts. Second fraction, which contains 23 second fraction extracts [Fr K1 to Fr. K23 Purifying the Fr·K7 of the second fraction extract by a normal phase high performance liquid chromatography system (NP-HPLC), and using a mixture of di-methane and ethyl acetate for the second preferred embodiment of the present invention. The white whip vinegar C of the example. The separation method of the two preferred embodiments described above can suitably extract the white whistle coral ester C and the white whit coral ester D of the first preferred embodiment and the second preferred embodiment of the present invention. Table 1. The biological activity of the preferred embodiment of the present invention is shown in the antioxidant activity [inhibit〇ry his (1) measurement results. Compound inhibits the percentage of superoxide anion production (O / 〇) a chlorpyrifos C -__H.61 + 2.80ca. Percent inhibition of oxidation in concentration _g / mL [%], its error van mean + SEM (n = 3 ] 0 9 1353355 .. •, Table 1 shows the biological activity of the white horn coral ester in the preferred embodiment of the present invention. The detection of super-anion anion production of human neutrophils in a concentration of 10 pg/mL [SUperoxide] Anion generation] The percentage of inhibition of superoxide anion formation by fMet-Leu-Phe/Cytochalastin B; wherein the white whip coral ester C has 11.61 ± 2.80%. The above experimental data are preliminary experimental results obtained under specific conditions. It is only for easy understanding or reference to the technical content of the present invention, and other experiments are required. The experimental data and the results thereof are not intended to limit the scope of the present invention. The foregoing preferred embodiment is merely illustrative. The invention and its technical features, the technology of this embodiment can be implemented by various substantial equivalent modifications and/or alternatives; therefore, the scope of the invention is subject to the appended claims. Please refer to the scope defined by the patent scope. 10 1353355 [Simple description of the drawings] Fig. 1 is a schematic view showing the structure of the chemical formula of the white whip coral ester c according to the first preferred embodiment of the present invention. A schematic structural diagram of the chemical formula of the white whip coral ester D of the preferred embodiment.
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