1286075 經濟部智慧財產局員工消費合作社印製 A7 B7 五、發明說明（1 ) 發明領域 本發明係關於一種具抗菌活性的醫藥組成物，其包含 從薑製備含抗菌活性的有效成分。 背景 在中國或日本，薑在食用或藥用方面都是很普遍的材 料。生薑是一般民眾日常生活中不可或缺的調味佐料，中 醫的臨床多用於發散風寒，健胃止嘔，解毒，薑汁可作爲 外敷，具有消炎止痛之效（臨床實用彩色科學中藥大典， 國際中文版第一冊，76頁，立得出版社）。在沙烏地阿拉伯， 薑則用於治療嘔吐、促進消化或作爲驅風劑（Ageel Μ α/.， 1 987, Plants Used in Saudi Folk Medicine, King Saud University Press，Riyadh，ρ·400) o 近年來，許多科學硏究顯示薑具有廣泛的生理活性。 例如：含薑的複方可抑制癌症轉移（日本專利公告號碼 0721 28 1 04)、抗病毒（日本專利公告號碼071 1 861)、輔助神 經營養因子生成，改善阿茲海默症與帕金森氏症等腦部疾 病（日本專利公告號碼07025777)、抗關節炎（日本專利公 告號碼06293653、美國專利第5494668號、美國專利第 5683698號）、抗微生物（日本專利公告號碼0622793 1)、鎭 ；» 痛（日本專利公告號碼061 07556)等。事實上，薑含1〜4 %揮 發油，其中約有200種物質被分離出來。在美國專利第 5804603號中提到，薑所含薑酮（zingerone)是辛辣味的主要 來源，且薑醇（gingerol)與生薑醇（shogaol)也具有辛香味。 ：-----1 —--------訂---------線 -------— ^------------- (往明閱讀背面之注項再填寫本頁) 1 2 本紙張尺度適用中國國家標準（CNS)A4規格（210 X 297公爱） 經濟部智慧財產局員工消費合作社印製 1286075 A7 __B7 五、發明說明（2 ) 此外，薑醇有強心、抑制老鼠離體靜脈收縮、調控老鼠與 兔子血管因eicosanoid引起的收縮等作用，而生薑醇則可抑 制血管收縮。在Srivastava與Mustafa的硏究中顯示薑可抑 制環氧化酵素（cyclooxygenase)與脂氧化酵素 (lipoxygenase)產物生成，同時抑制eicosanoid的合成 (Medical Hypotheses, 1 992, 39: 342-348)。Mascolo N·等人 則發現，在以100 mg/kg的劑量口服給藥後，薑之80 %酒精 萃取物與乙醢水楊酸（acetylsalicylic acid)對鹿角菜膠 (carageenan)誘發的老鼠腳掌浮腫的抑制效果相當，抑制率 分別爲39 %與38 %。在同一硏究中也發現，薑之80%酒精 萃取物可抑制酵母所引起的發熱，並可抑制格蘭氏陰性菌 與格蘭氏陽性菌的生長（Journal of Ethnopharmocology， 1 989，27，129_140)。Srivastava 與 Mustafa 以薑粉末治療 56 個臨床病人，結果顯示四分之三的關節炎病人減輕疼痛或 腫脹，而所有肌肉不適的病人都感覺疼痛減輕，而在三個 月到兩年半的臨床期間，沒有病人表示有不良效果產生 (1992, ibid)。 皮癣菌（Dermaiop/iyiesj，尤其紅色髮癬菌 (Trichophyton rubrum)^M^M MMW ( Trichophyton mentagrophytes)，爲甲黴菌病（onychomycosis)及足癬 (tinea pedis)的通常病原[Roberts DT·，British Journal of Dermatology. 141 Supple 56:1-4, 1999 Nov.; Roldan YB. et al·，Mycoses，43(5):181-3, 2000]。卵狀糠疹癬菌 [P/iyrospori/n7 o\/a/e (/Wa/assez/a 爲花斑癬 5 本紙張尺度適用中國國家標準（CNS)A4規格（210 χ 297公釐） ---------II — — — — — — — I— « — — — — — — II I Aw----------^------------- (請先閱讀背面之注音？事項再填寫本頁) 經濟部智慧財產局員工消費合作社印製 1286075 A7 __B7_ 五、發明說明（3 ) (pityriasis vesicolor)、皮膚芽胞菌濾胞炎（P/iyrosporun7 folliculitis)及馬辣塞氏菌擦瘆（Ma/assez/a intertrig〇)的病 因。已有數個硏究顯示卵狀糠疹癬菌o\/a/e) 與皮脂性皮膚炎（seborrheic dermatitis)及頭皮屑有關（頭 皮屑屬於一種較輕症狀的皮脂性皮膚炎）[Nenoff P. et al., Dermatology. 191 (4):311-4, 1 995; Bulmer AC. et al., Mycopathologia, 147(2)63-5, 1 999]° 發明要旨 本發明是利用有機溶劑（乙醚、丙酮、甲醇、乙醇）或超 臨界二氧化碳萃取，或以水蒸氣蒸餾從薑 的根莖準備一粗製液，再經逆相層析法精製而得 到含有生薑醇（shogaols)、薑醇（gingerols)及/或去氫薑二酮 (dehydrogingerdione)的產物。這些產物經體外抗菌活性試 驗證實具有明顯的抗菌活性作用。 發明詳述 如背景所述，薑汁有消炎、止痛效用之記載。 本發明旨在提供一種有效的方法從薑的根莖製備出具 抗菌活性的有效成分的產品，並且所製得的產品具有實質 上固定的等有效成份含量，於是亦具有明確的藥理作用。 依本發明內容所完成的一種從薑製備含抗茵的有效成 分的產品方法，包含下列步驟： a)從薑（ZMgz·心r 的根莖製備一粗製液； 6 本紙張尺度適用中國國家標準（CNS)A4規格（210 X 297公釐） ----- • ^1 ϋ n ϋ ϋ ϋ 一口V a n ϋ ϋ 1 I 線 —φ------.---·------------- (請先閱讀背面之注意事項再填寫本頁) 經濟部智慧財產局員工消費合作社印製 1286075 A7 __B7_____ 五、發明說明（4 ) b) 將該粗製液導入一逆相層析管柱，並依序以水、第 一流洗液及第二流洗液流洗，收集其中的第一流洗液的溶 離物及第二流洗液的溶離物，其中該第一流洗液的極性大 於該第二流洗液，該第二流洗液的極性大於氯仿；及 c) 揮發移除該第一流洗液的溶離物中的第一流洗液而 得一第一濃縮溶離物，該第一濃縮溶離物可作爲該產品被 取出；及 d) 揮發移除該第二流洗液的溶離物中的第二流洗液而 得一第二濃縮溶離物，該第二濃縮溶離物可作爲該產品被 取出， 其中該步驟a)包含下列步驟i)至iv)，步驟I)，步驟Γ) 或步驟I”），該步驟i)至iv)爲： i) 將薑的新鮮根莖粉碎及過濾，而 得到一濾液及一殘渣部份； ii) 將該濾液以一第一有機溶劑萃取，取出所獲得的第 一有機溶劑萃取液，並揮發移除其中的第一有機溶劑而獲 得一濃縮的第一有機溶劑萃取液； iii) 將該殘渣部份以一第二有機溶劑萃取’取出所獲得 的第二有機溶劑萃取液，並揮發移除其中的第二有機溶劑 而獲得一濃縮的第二有機溶劑萃取液；及 iv) 合倂該濃縮的第一有機溶劑萃取液及該濃縮的第 二有機溶劑萃取液而獲得該粗製液； 該步驟I)爲： I)以該第二有機溶劑萃取被粉碎的薑 7 本紙張尺度適用中國國家標準（CNS)A4規格（210 X 297公釐） y 一一口V ϋ ϋ n ϋ n I ϋ n ·1 ϋ ϋ ϋ ϋ ϋ n ϋ ·ϋ 1 ϋ n ϋ ϋ I - (請先閱讀背面之注意事項再填寫本頁) 1286075 A7 B7 五、發明說明（5 ) 的乾燥根莖，取出所獲得的第二有機溶劑萃取 液，並揮發移除其中的第二有機溶劑而獲得該粗製液； 該步驟Γ)爲： Γ)以水蒸氣蒸餾被粉碎的薑（ZhgAer 的乾 燥根莖，減壓濃縮所獲得的餾出物而獲得該粗製液；及 該步驟Γ)爲： Γ)以超臨界二氧化碳萃取被粉碎的薑 以7/^/7α/〇的乾燥根莖，並揮發移除其中的二氧化碳而獲得 該粗製液。 依本發明方法所製備的該含抗菌有效成分的產品包含 0 - 10 mg/g 的生薑醇（shogaol)，1 - 150 mg/g 的薑醇（gingerol) 及 0 - 40 mg/g 的 6-去氫薑二酮（6-dehydrogingerdione)。 本發明亦提供一種具抗菌活性的醫藥組成物，包含一 抗菌治療有效量的依前述本發明方法的步驟a)所製備的粗 製液作爲有效成分，及醫藥上可接受之載體或稀釋劑。 本發明亦提供一種具抗菌活性的醫藥組成物，包含依 前述本發明方法所製備的產品作爲有效成分，及醫藥上可 接受之載體或稀釋劑。較佳地，該產品爲該第一濃縮溶離 物。選擇性地，該產品爲該第二濃縮溶離物。 較佳地，該第一流洗液爲甲醇及該第二流洗液爲丙 酮。 較佳地，該步驟a)包含步驟i)至iv)。 較佳地，該第一有機溶劑爲乙醚。 較佳地，該第二有機溶劑爲丙酮、甲醇、乙醇或其等 8 本紙張尺度適用中國國家標準（CNS)A4規格（210 X 297公釐） (請先閱讀背面之注意事項再填寫本頁)1286075 Ministry of Economic Affairs Intellectual Property Office Employees Consumption Cooperative Printed A7 B7 V. INSTRUCTION DESCRIPTION (1) Field of the Invention The present invention relates to a pharmaceutical composition having antibacterial activity, which comprises preparing an active ingredient containing antibacterial activity from ginger. Background In China or Japan, ginger is a very common material in terms of consumption or medicinal use. Ginger is an indispensable seasoning in the daily life of the general public. The clinical practice of traditional Chinese medicine is mostly used for divergent cold, stomach and vomiting, detoxification, ginger juice can be used as external application, with anti-inflammatory and analgesic effects (clinical practical color science Chinese medicine ceremony, International Chinese Edition, Volume 1, 76 pages, Lied Publishing House). In Saudi Arabia, ginger is used to treat vomiting, promote digestion or act as a wind-driven agent (Ageel Μ α/., 1 987, Plants Used in Saudi Folk Medicine, King Saud University Press, Riyadh, ρ·400) o Many scientific studies have shown that ginger has a wide range of physiological activities. For example, a compound containing ginger can inhibit cancer metastasis (Japanese Patent Publication No. 0721 28 1 04), antiviral (Japanese Patent Publication No. 071 1 861), helper neurotrophic factor production, and improve Alzheimer's disease and Parkinson's disease. And other brain diseases (Japanese Patent Publication No. 07025777), anti-arthritis (Japanese Patent Publication No. 06239653, US Patent No. 5494668, US Patent No. 5683698), Antimicrobial (Japanese Patent Publication No. 0622793 1), 鎭; (Japanese Patent Publication No. 061 07556) and the like. In fact, ginger contains 1 to 4% of the oil, of which about 200 substances are separated. As mentioned in U.S. Patent No. 5,804,603, ginger contains zingerone which is the main source of spicy taste, and gingerol and shogaol also have a pungent aroma. :-----1 —--------Book --------- Line-------- ^------------- ( Please read the note on the back of the page and fill in this page. 1 2 This paper scale applies to China National Standard (CNS) A4 specification (210 X 297 public). Ministry of Economic Affairs Intellectual Property Bureau employee consumption cooperative prints 1286075 A7 __B7 V. Invention Description (2) In addition, gingerol has a strong heart, inhibits the contraction of isolated veins in mice, regulates the contraction of blood vessels caused by eicosanoid in mice and rabbits, and ginger alcohol can inhibit vasoconstriction. In the study of Srivastava and Mustafa, it was shown that ginger inhibits the production of cyclooxygenase and lipoxygenase, while inhibiting the synthesis of eicosanoid (Medical Hypotheses, 1 992, 39: 342-348). Mascolo N· et al. found that after oral administration at a dose of 100 mg/kg, 80% alcohol extract of ginger and acetylsalicylic acid on carageenan-induced paw swelling in mice The inhibitory effect was comparable, and the inhibition rates were 39% and 38%, respectively. In the same study, it was also found that 80% alcohol extract of ginger inhibited the fever caused by yeast and inhibited the growth of Gram-negative bacteria and Gram-positive bacteria (Journal of Ethnopharmocology, 1 989, 27, 129_140). ). Srivastava and Mustafa treated 56 clinical patients with ginger powder. The results showed that three-quarters of arthritis patients relieved pain or swelling, while all patients with muscle discomfort felt pain relief during the three-month to two-and-a-half-year clinical period. No patient indicated adverse effects (1992, ibid). Dermaiop/iyiesj, especially Trichophyton rubrum^M^M MMW (Trichophyton mentagrophytes), a common pathogen for onychomycosis and tinea pedis [Roberts DT·, British Journal of Dermatology. 141 Supple 56:1-4, 1999 Nov.; Roldan YB. et al., Mycoses, 43(5): 181-3, 2000]. E. falciparum [P/iyrospori/n7 o \/a/e (/Wa/assez/a is the tinea 癣5 This paper scale applies to the Chinese National Standard (CNS) A4 specification (210 297 297 mm) ---------II — — — — — — — I— « — — — — — — II I Aw----------^------------- (Please read the phonetic on the back first? This page) Ministry of Economic Affairs Intellectual Property Office Staff Consumer Cooperative Printed 1286075 A7 __B7_ V. Description of invention (3) (pityriasis vesicolor), cutaneous spore cytitis (P/iyrosporun7 folliculitis) and horse serrata rubbing (Ma/ The cause of assez/a intertrig〇). Several studies have shown that Ophiopogon japonicus o\/a/e) is associated with seborrheic dermatitis and dandruff (dandruff belongs to a kind) Symptoms of sebaceous dermatitis) [Nenoff P. et al., Dermatology. 191 (4): 311-4, 1 995; Bulmer AC. et al., Mycopathologia, 147(2)63-5, 1 999]° SUMMARY OF THE INVENTION The present invention utilizes an organic solvent (diethyl ether, acetone, methanol, ethanol) or supercritical carbon dioxide extraction, or steam distillation to prepare a crude liquid from the roots of ginger, and then refined by reverse phase chromatography to obtain ginger. The product of shogaols, gingerols and/or dehydrogingerdione. These products have been shown to have significant antibacterial activity by in vitro antibacterial activity tests. DETAILED DESCRIPTION OF THE INVENTION As background, ginger juice has Description of the anti-inflammatory and analgesic effects. The present invention aims to provide an effective method for preparing an antibacterial active ingredient from the roots of ginger, and the obtained product has a substantially fixed active ingredient content, and thus is also clear Pharmacological effects. A method for preparing a product containing an antibacterial active ingredient from ginger according to the present invention comprises the following steps: a) preparing a crude liquid from the root of the ginger (ZMgz·heart r; 6) The paper scale is applicable to the Chinese national standard ( CNS)A4 size (210 X 297 mm) ----- • ^1 ϋ n ϋ ϋ ϋ One mouth V an ϋ ϋ 1 I line—φ------.--------- -------- (Please read the note on the back and then fill out this page) Ministry of Economic Affairs Intellectual Property Bureau employee consumption cooperative printed 1286075 A7 __B7_____ V. Invention description (4) b) Introduce the crude liquid into a counter The phase chromatography column is sequentially washed with water, the first stream washing liquid and the second stream washing liquid, and the first flowing washing liquid dissolving material and the second flowing washing liquid dissolving material are collected, wherein the first flow washing The polarity of the liquid is greater than the second stream washing liquid, the second stream washing liquid is more polar than chloroform; and c) volatilizing to remove the first stream washing liquid in the first flowing lotion to obtain a first concentrated dissolving material The first concentrated dissolvate may be taken out as the product; and d) volatilized to remove the second flowing lotion in the eluate of the second flowing lotion a second concentrated lysate, the second concentrated lysate may be taken out as the product, wherein the step a) comprises the following steps i) to iv), step I), step Γ) or step I"), the step i ) to iv): i) crushing and filtering the fresh rhizome of ginger to obtain a filtrate and a residue fraction; ii) extracting the filtrate as a first organic solvent, and taking out the obtained first organic solvent extract And volatilizing the first organic solvent to obtain a concentrated first organic solvent extract; iii) extracting the residue portion with a second organic solvent to extract the obtained second organic solvent extract, and Volatilizing to remove the second organic solvent to obtain a concentrated second organic solvent extract; and iv) combining the concentrated first organic solvent extract and the concentrated second organic solvent extract to obtain the crude liquid This step I) is: I) extraction of the pulverized ginger with the second organic solvent 7 The paper size is applicable to the Chinese National Standard (CNS) A4 specification (210 X 297 mm) y One-to-one V ϋ ϋ n ϋ n I ϋ n ·1 ϋ ϋ ϋ ϋ ϋ n ϋ ·ϋ 1 ϋ n ϋ ϋ I - (Please read the notes on the back and fill out this page) 1286075 A7 B7 V. Drying rhizome of invention description (5), take out the obtained second organic solvent extract, and volatilize and remove it. The second organic solvent is used to obtain the crude liquid; the step Γ) is: Γ) steaming the pulverized ginger (dry root of ZhgAer, distilling the obtained distillate under reduced pressure to obtain the crude liquid; and the step) Γ) is: Γ) The dried rhizome of 7/^/7α/〇 is extracted with supercritical carbon dioxide by supercritical carbon dioxide, and the carbon dioxide is removed by volatilization to obtain the crude liquid. The antibacterial active ingredient-containing product prepared by the method of the present invention comprises 0 - 10 mg/g of gingerol (shogaol), 1 - 150 mg/g of gingerol (gingerol) and 0-40 mg/g of 6 - Dehydrogenated dione (6-dehydrogingerdione). The present invention also provides a pharmaceutical composition having antibacterial activity comprising an antibacterial therapeutically effective amount of the crude liquid prepared according to the step a) of the above method of the present invention as an active ingredient, and a pharmaceutically acceptable carrier or diluent. The present invention also provides a pharmaceutical composition having antibacterial activity comprising the product prepared by the method of the present invention as an active ingredient, and a pharmaceutically acceptable carrier or diluent. Preferably, the product is the first concentrated lysate. Optionally, the product is the second concentrated dissolvate. Preferably, the first stream washing liquid is methanol and the second stream washing liquid is acetone. Preferably, step a) comprises steps i) to iv). Preferably, the first organic solvent is diethyl ether. Preferably, the second organic solvent is acetone, methanol, ethanol or the like. The paper size is applicable to the Chinese National Standard (CNS) A4 specification (210 X 297 mm). (Please read the back note first and then fill in the page. )
---------訂———•線J 經濟部智慧財產局員工消費合作社印製 1286075 A7 ___B7___ 五、發明說明（6 ) 之混合。更佳地，該第二有機溶劑爲丙酮。 較佳地，該步驟a)包含步驟I)。 (請先閱讀背面之注意事項再填寫本頁) 選擇性地，該步驟a)包含步驟Γ)。 選擇性地，該步驟a)包含步驟Γ)。 適用於本發明方法的逆相層析管柱包括（但不限於）被 塡充有多孔性聚合物的樹脂，例如Diaion HP-20 (三菱公 司 Mitsubishi Co.)、Sephadex LH-20 (Pharmicia Co.)或 RP-18 (Nacalai tesque Co.)等逆相層析樹脂。 本發明的具抗菌活性的醫藥組成物較佳的係局部地被 施用例如以洗髮精、沐浴乳、肥皂、身體乳液、身體乳霜 及洗潔劑的型式被使用。較佳的，被用於治療與髮瘡小芽 孢癬菌（7>/(；/7〇厂/7)^〇^7^70以39^〇0/7/化$)及卵狀糠疹癬菌 (P/iyrospooym 有關的疾病，包括但不限於足癬 經濟部智慧財產局員工消費合作社印製 (tinea pedis)、髮癬（tinea capitis)、腹股溝癬（tinea cruris)、滑癬（tinea glabrosa)、甲黴菌病 (onychomycosis)、頭皮慷癖（pityriasis capms)、花斑癖 (pityriasis vesicolor)、皮膚芽胞菌濾胞炎（P/fyrospori/m folliculitis)、皮脂性皮膚炎（seborrheic dermatitis)、及頭 皮屑（dand ruff)。尤其本發明的具抗菌活性的醫藥組成物爲 一用於治療頭皮屑的洗髮精型式。 本發明將藉以下實施例被進一步瞭解，該等實施例僅 爲說明之用而非用於限制本發明範圍。 有效成分之測定 於以下貫施例中使用局效液相層析（high performance 9 本紙張尺度適用中國國家標準（CNS)A4規格（210 X 297公釐） A7 1286075 B7 五、發明說明（7 ) liquid chromatography，簡稱HPLC)來測定本發明方法所製 備的產品的有效成分。HPLC光譜係被記錄於一高效液相層 析儀（HPLC Shimadzu LC-10AT，Japan)，其中適量的樣品以 移動相溶液[甲腈與水（65 : 3 5，V/V)的混合液]定量至25 ml，經0.25 μιη濾膜過濾後，將濾液注入一塡充有Cosmosil 5C-18的管柱（長25 cm，內徑4·6 mm，粒子大小5 μιη)，以 該移動相溶液進行流洗（elution)。採用紫外光偵測器 (Shimadzu SPD-6AV, Japan)，偵測溶離液（eluate)在波長 230 nm的吸收。 實施例一 將生薑2100 g打碎後過濾得濾液與薑殘渣。取500 ml 濾液以500 ml乙醚萃取三次，將每次萃取得到的水層與乙 醚層分離，並合倂被分離的乙醚層，揮發移除其中的乙醚 而得乙醚層濃縮物（I-OE)。另將薑殘渣以3000 ml丙酮萃取 三次，過濾後得丙酮萃取液，將每次萃取得到丙酮萃取液 合倂並減壓濃縮得丙酮抽出物（1-0) 14.5 g。將乙醚層濃縮 物（I-OE)與丙酮抽出物（1-0)合倂後取7 g，注入一塡充有 180 g Diaion HP-20樹脂的逆相色層層析管柱（長度30 cm， 內徑3 cm，粒徑 500 μητι-800 μηη)，依序以 1500 ml水、2500 ml甲醇、2000 ml丙酮與2000 ml氯仿進行流洗（elution)，收 集水溶離物、甲醇溶離物、丙酮溶離物與氯仿溶離物，減 壓濃縮後得濃縮水溶離物（I-OW) 0.27 g、濃縮甲醇溶離物 (I-OM) 1.45 g、濃縮丙酮溶離物（I-OA) 2.68 g與濃縮氯仿 10 本紙張尺度適用中國國家標準（CNS)A4規格（210 X 297公釐） (請先閱讀背面之注意事項再填寫本頁) 經濟部智慧財產局員工消費合作社印製 -1111111 I — — — — — — — — I Αν I I I I I I- —1!^— — — — — — — — — — — — — . 1286075 A7 B7 五、發明說明（8 ) 溶離物（I-OC) 0.83 g。其中’ 1-〇、km與I-OA中6 -生薑醇、 6-薑醇與6 -去氫薑二酮的含量如袠一所示。 表〜 含量 (mg/g) 1-0 Ι-ΟΜ Ι-ΟΑ 6-生薑醇 1·10±0·14 1.15±0.0 — 6-薑醇 59.98±0.99 103.37±8.57 2·51±0·89 6-去氫薑二酮 7.68±0.42 8.94±0.41 ' - ----·----1----___ (請先閱讀背面之注意事項再填寫本頁) 將乾薑500 g打碎後直接以30 L丙酮分三次萃取（每次 1 0 L)，過濾後得丙酮濾液，合倂濾液並減壓濃縮得丙酮抽 出物（11-0) 24 g。取20 g丙酮抽出物，通過塡充600 g Diaion HP-20樹脂的逆相色層層析管柱，依序以4 L水、6.5 L曱 醇、1 5 L丙酮與5 L氯仿流洗，收集水溶離物、甲醇溶離物、 丙酮溶離物與氯仿溶離物，分別減壓濃縮後得濃縮水溶離 物（II-OW) 2.5 g、濃縮甲醇溶離物（ΙΙ-ΟΜ) 7·1 g、濃縮丙 酮溶離物（Π-ΟΑ) 6.9 g與濃縮氯仿溶離物（II-OC) 3.5 g。其 中，11-0、II-OM與II-OA中6-生薑醇、6-薑醇與6-去氫薑二 酮的含量如表二所示。 11 本紙張尺度適用中國國家標準（CNS)A4規格（210 X 297公釐） I 訂 經濟部智慧財產局員工消費合作社印製 1286075 A7 五、發明說明（9 ) 表二 含量 — (mg/g) 11-0 II-OM ΙΙ-ΟΑ 6-生薑醇 1.98±0.00 4.96±0.00 一 6-薑醇 43.06±0.84 70.87±1.85 2.5410.00 _ 6-去氫薑二酮 9.33±0.85 19·15±4·57 2.35±0.28 實施例三 將乾薑10 kg打碎後以水蒸氣蒸餾5小時，將餾出物減壓 濃縮得蒸餾物（ΙΙΙ-0) 410 g。取20 g蒸餾物通過塡充600 g Diaion HP-20樹脂的逆相色層層析管柱，依序以4.5 L水、 4·5 L甲醇、3 L丙酮與5 L氯仿流洗，收集水溶離物、甲醇 溶離物、丙酮溶離物與氯仿溶離物，減壓濃縮後得濃縮水 溶離物（III-OW) 0.03 g、濃縮甲醇溶離物（ΙΙΙ_ΟΜ) 14.5 g、 濃縮丙酮溶離物（III-OA) 0.8 5 g與濃縮氯仿溶離物（III-OC) 〇·2 g。乾薑之水蒸氣蒸餾物（ΙΙΙ-0)經HPLC分析不含6-生薑 醇、6-薑醇與6·去氫薑二酮。 實施例四 取10 g粉碎乾薑以1000 ml丙酮於50°C萃取2小時，過濾 除去植物殘體，減壓濃縮後以真空乾燥（4(TC，75 mmHg真 空度）得丙酮萃取物IV-O。產率與產物的色澤及黏度見表 —- 〇 12 本紙張尺度適用中國國家標準（CNS)A4規格（210 X 297公釐） ----\——l·——ΛΨ (請先閱讀背面之注意事項再填寫本頁) 經濟部智慧財產局員工消費合作社印製 訂---------線_ •-----,.i!r------------- 1286075 A7 B7 五、發明說明（1〇 ) 實施例五 取1 〇 g粉碎乾薑以水蒸氣蒸餾，分別將其油溶性與水溶 性萃取部分冷凍乾燥，得油溶性萃取物V-0與水溶性萃取 物ν-w。產率與產物的色澤及黏度見表三。 實施例六 取10 g粉碎乾薑置於體積25 0 ml之萃取槽中，以高壓幫 浦（LEWA公司EK-1型，美國）控制超臨界C〇2壓力，設定 壓力於2500至4000 psia，溫度由熱交換機（HOTEC公司 H-24 10型，台灣）配合萃取槽外循環管路控制，維持操作溫 度於35-60°C，流速控制於45 L/min，當(：02體積達到300 L 時，停止萃取步驟，得萃取物VI-0。產率與產物的色澤及 黏度見表三，產物的辣味成份含量見表四。 表三 IV-0 V-0 VI-0 L* 87.6 80.4 96.3 A* -9.1 -0.1 -9.6 B* 31.1 9.6 22.0 黏度(cPs) 15.6 11.8 12.1 產率(％) 3.8 2.2 3.9 * : L、A、B之値係以色差計（Σ90 color measuring system， Nippon Denshoku Inc，co·，LTD, Japan)測量，明度以L表 示，色度以A與B表示。 13 本紙張尺度適用中國國家標準（CNS)A4規格（210 X 297公釐） -I---^——l·--- (請先閱讀背面之注音S事項再填寫本頁) 經濟部智慧財產局員工消費合作社印製 ^1 ^1 ^1 ϋ ϋ I ϋ ϋ ϋ Βϋ ϋ ^1 ^1 ^1 ^1 ^1 ϋ ϋ I I I ^1 «ϋ ϋ I ϋ 1286075 A7 B7 五、發明說明（11 )表四 含量(mg/g) VI-0 6-生薑醇 17.30±0.00 6-薑醇 26.29±0.00 6-去氫薑二酮 19.20±1.19 經濟部智慧財產局員工消費合作社印製 實施例七抑制髮瘡小芽孢癬菌（⑽及卵狀糠痊癬菌瓜 之活性評估對髮瘡小芽孢癬菌抑制作用的評估 薑萃取物對髮瘡小芽孢癬菌（Trichophyton 11^1^&81*〇卩11>^68;八丁(1^ 95 3 3)的抑制作用根據£(1'\¥&1(13，】.11· 等人的方法加以評估（Edwards, J· R· et al·，Antimicrobial Agents Chemotherapy 33: 21 5-222，1989)，並以最小抑菌濃 度表示（minimum inhibitory concentration; MIC)，將臺萃取 物以二甲亞楓（dimethyl sulfoxide，簡稱DMS0)溶解後再 稀釋成各種所需濃度的基本溶液（stock solution)。將0.01 毫升基本溶液加入含有0.99毫升馬鈴薯葡萄糖營養液 [Potato Dextrose Broth (DIFCO, U.S.A·)]與 103 至 104CFU/ml髮瘡小芽孢癬菌的48孔培養盤中，培養盤在28 °C下溫浴72小時後以肉眼檢視決定萃取物的抑制作用，實 驗中並以溶媒作爲空白對照組，每一試驗皆以二重覆的方 式進行。結果如表五所示。 14 本紙張尺度適用中國國家標準（CNS)A4規格（210 X 297公釐） -----^----L---- (請先閱讀背面之注意事項再填寫本頁) 訂--------- 1286075 A7 __B7_____ 五、發明說明（12 ) 對卵狀糠疹癬菌的抑制作用 薑萃取物對卵狀糠瘆癖菌（Pityrosporum ovale ; ATCC 38593> 的抑制作用亦根據上述Edwards，J. R.等人的方法加以評 估，並以最小抑菌濃度表示（minimum inhibitory concentration; MIC)，將薑萃取物以DMSO溶解後再稀釋成所需濃度的基 本溶液。將0.01毫升基本溶液加入含有0.99毫升流體狀 薩蒲洛氏媒體中[Fluid Sabouraud Medium (DIFCO, U .S. A·)]與 1 03 -104 CFU/ml卵狀糠疹廯菌的48孔培養盤中，於37°C下溫 浴48小時後，以肉眼檢查決定萃取物的抑制作用，實驗中 並以溶媒DMSO作爲空白對照組，每一試驗皆以二重覆的 方式進行。結果如表五所示。 表五薑萃取物對卵狀糠疹癬菌和髮瘡小芽孢癖菌的抑制 作用 薑萃取物a) MIC (pg/ml) 卵狀糠疹癬菌 髮瘡小芽孢癬菌 1-0 100 30 I-OM 100 30 11-0 500 30 II-OC 100 30 II-OM 100 30 ΙΙΙ-0 500 100 空白對照組 __ b) _ b) 於實施例一、二及三所製備者 無抑制作用 15 本紙張尺度適用中國國家標準（CNS)A4規格（210 X 297公釐） (請先閱讀背面之注意事項再填寫本頁) --------訂---------線» 經濟部智慧財產局員工消費合作社印製---------Booking ---• Line J Ministry of Economic Affairs Intellectual Property Bureau employee consumption cooperative printing 1286075 A7 ___B7___ Five, a description of the invention (6). More preferably, the second organic solvent is acetone. Preferably, step a) comprises step I). (Please read the notes on the back and fill out this page.) Optionally, step a) contains the steps Γ). Optionally, step a) comprises the step Γ). Reverse phase chromatography columns suitable for use in the method of the invention include, but are not limited to, resins that are filled with a porous polymer, such as Diaion HP-20 (Mitsubishi Co., Mitsubishi Co.), Sephadex LH-20 (Pharmicia Co.). Or reverse phase chromatography resin such as RP-18 (Nacalai tesque Co.). The antibacterial active pharmaceutical composition of the present invention is preferably applied topically, for example, in the form of shampoo, body wash, soap, body lotion, body cream and detergent. Preferably, it is used for the treatment of acne spores (7>/(;/7〇厂/7)^〇^7^70 to 39^〇0/7/$) and ovate-like pityriasis癣 ( (P/iyrospooym related diseases, including but not limited to the 癣 癣 癣 癣 员工 员工 员工 ti ti ti ti ti ti ti ti ti ti ti ti ti ti ti ti ti ti ti ti ti ti ti ti ti ti ti ti ti ti ti ti ti ti ti ti ti ti ti ti ti ti ti ti ti ti ti ti ti ti ti ti ), onychomycosis, pityriasis capms, pityriasis vesicolor, P/fyrospori/m folliculitis, seborrheic dermatitis, and scalp Dand ruff. In particular, the antibacterial active pharmaceutical composition of the present invention is a shampoo type for treating dandruff. The present invention will be further understood by the following examples, which are for illustrative purposes only. Rather than limiting the scope of the invention. Determination of active ingredients is carried out in the following examples using high performance liquid chromatography (high performance 9 paper scale applicable to Chinese National Standard (CNS) A4 specification (210 X 297 mm) A7 1286075 B7 V. Description of invention (7 ) liquid Chromatography (referred to as HPLC) to determine the active ingredient of the product prepared by the method of the present invention. The HPLC spectrum was recorded on a high performance liquid chromatography (HPLC Shimadzu LC-10AT, Japan), in which the appropriate amount of the sample was in the mobile phase solution [ A mixture of carbonitrile and water (65: 3 5, V/V) was quantified to 25 ml, filtered through a 0.25 μm filter, and the filtrate was poured into a column packed with Cosmosil 5C-18 (length 25 cm, The inner diameter is 4·6 mm, the particle size is 5 μιη), and the mobile phase solution is subjected to elution. The ultraviolet light detector (Shimadzu SPD-6AV, Japan) is used to detect the elution solution (eluate) at a wavelength of 230. Absorption of nm. In Example 1, 2100 g of ginger was crushed and filtered to obtain a filtrate and a ginger residue. 500 ml of the filtrate was extracted three times with 500 ml of diethyl ether, and the aqueous layer obtained by each extraction was separated from the ether layer and combined. The separated ether layer was evaporated to remove the diethyl ether to obtain a diethyl ether layer concentrate (I-OE). The ginger residue was extracted three times with 3000 ml of acetone, and filtered to obtain an acetone extract, and each extraction was carried out to obtain an acetone extract. The mixture was concentrated under reduced pressure to give acetone extract (1-0), 14.5 g. The diethyl ether layer concentrate (I-OE) was combined with the acetone extract (1-0), and 7 g was taken, and a reverse phase chromatography column (180 cm long) filled with 180 g of Diaion HP-20 resin was injected. , inner diameter 3 cm, particle size 500 μητι-800 μηη), sequentially elution with 1500 ml water, 2500 ml methanol, 2000 ml acetone and 2000 ml chloroform, collecting water-soluble, methanol-solvent, acetone The dissolved material and the chloroform solution were concentrated under reduced pressure to obtain a concentrated water-soluble (I-OW), 0.27 g, concentrated methanol (I-OM), 1.45 g, concentrated acetone (I-OA), 2.68 g, and concentrated chloroform. 10 The paper size applies to the Chinese National Standard (CNS) A4 specification (210 X 297 mm) (please read the note on the back and then fill out this page) Ministry of Economic Affairs Intellectual Property Bureau Staff Consumer Cooperative Printed -1111111 I — — — — — — — — I Αν IIIII I- —1!^— — — — — — — — — — — — — 1286075 A7 B7 V. INSTRUCTIONS (8) Dissolution (I-OC) 0.83 g. The contents of 6-glycol alcohol, 6-gingerol and 6-dehydroglycinedion in '1-〇, km and I-OA are as shown in 袠. Table ~ Content (mg/g) 1-0 Ι-ΟΜ Ι-ΟΑ 6-Gingerol 1·10±0·14 1.15±0.0 — 6-Gingerol 59.98±0.99 103.37±8.57 2·51±0·89 6-Dehydrogenated dione 7.68±0.42 8.94±0.41 ' - ----·----1----___ (Please read the note on the back and fill in this page) Break the dried ginger 500 g After that, it was directly extracted with 30 L of acetone three times (10 L each time), and the acetone filtrate was obtained by filtration, and the filtrate was combined and concentrated under reduced pressure to give acetone extract (11-0) 24 g. 20 g of acetone extract was taken and washed by a reverse phase chromatography column packed with 600 g of Diaion HP-20 resin, followed by 4 L of water, 6.5 L of decyl alcohol, 15 L of acetone and 5 L of chloroform. The water-soluble, methanol-solvent, acetone-soluble and chloroform-dissolved materials were collected, and concentrated under reduced pressure to obtain a concentrated water-soluble (II-OW) 2.5 g, concentrated methanol-soluble (ΙΙ-ΟΜ) 7·1 g, concentrated. Acetone dissolvate (Π-ΟΑ) 6.9 g and concentrated chloroform dissolvate (II-OC) 3.5 g. Among them, the contents of 6-gingerol, 6-gingerol and 6-dehydroglycinedione in 11-0, II-OM and II-OA are shown in Table 2. 11 The paper size is applicable to China National Standard (CNS) A4 specification (210 X 297 mm) I. Ministry of Economic Affairs Intellectual Property Office Staff Consumer Cooperative Printed 1286075 A7 V. Invention Description (9) Table 2 Content - (mg/g) 11-0 II-OM ΙΙ-ΟΑ 6-glycan alcohol 1.98±0.00 4.96±0.00 a 6-gingerol 43.06±0.84 70.87±1.85 2.5410.00 _ 6-dehydro gingerdione 9.33±0.85 19·15±4 · 57 2.35 ± 0.28 In the third embodiment, 10 kg of dried ginger was crushed and distilled by steam for 5 hours, and the distillate was concentrated under reduced pressure to obtain a distillate (ΙΙΙ-0) 410 g. 20 g of distillate was passed through a reverse phase chromatography column packed with 600 g of Diaion HP-20 resin, and sequentially washed with 4.5 L of water, 4.5 L of methanol, 3 L of acetone and 5 L of chloroform to collect water. Eluent, methanol dissolvate, acetone dissolvate and chloroform dissolvate, concentrated under reduced pressure to obtain concentrated water soluble (III-OW) 0.03 g, concentrated methanol dissolved (ΙΙΙ_ΟΜ) 14.5 g, concentrated acetone dissolvate (III-OA 0.8 5 g with concentrated chloroform dissolvate (III-OC) 〇·2 g. The steam distillate of dried ginger (ΙΙΙ-0) was analyzed by HPLC without 6-gingerol, 6-gingerol and 6·dehydroglycanedione. Example 4 10 g of pulverized dried ginger was extracted with 1000 ml of acetone at 50 ° C for 2 hours, and the plant residue was removed by filtration, concentrated under reduced pressure, and dried under vacuum (4 (TC, 75 mmHg vacuum) to give acetone extract IV- O. Yield and color and viscosity of the product are shown in the table - 〇12 This paper scale applies to China National Standard (CNS) A4 specification (210 X 297 mm) ----\——l·——ΛΨ (please first Read the notes on the back and fill out this page.) Ministry of Economic Affairs, Intellectual Property Bureau, Staff and Consumer Cooperatives, Printed---------Line _ •-----,.i!r-------- ----- 1286075 A7 B7 V. Inventive Note (1〇) Example 5 Take 1 〇g crushed dried ginger by steam distillation, and separately lyophilize the oil-soluble and water-soluble extract portions to obtain oil-soluble extract V -0 and water-soluble extract ν-w. Yield and product color and viscosity are shown in Table 3. Example 6 10 g of crushed ginger placed in a volume of 25 0 ml extraction tank, with high pressure pump (LEWA company EK-1 type, USA) Control supercritical C〇2 pressure, set pressure at 2500 to 4000 psia, temperature is controlled by heat exchanger (HOTEC H-24 10, Taiwan) with extraction tank Circulating line control, maintaining the operating temperature at 35-60 ° C, the flow rate is controlled at 45 L / min, when (: 02 volume reaches 300 L, the extraction step is stopped, the extract VI-0 is obtained. The yield and the color of the product The viscosity and the viscosity are shown in Table 3. The content of the spicy ingredients of the product is shown in Table 4. Table III IV-0 V-0 VI-0 L* 87.6 80.4 96.3 A* -9.1 -0.1 -9.6 B* 31.1 9.6 22.0 Viscosity (cPs) 15.6 11.8 12.1 Yield (%) 3.8 2.2 3.9 * : The enthalpy of L, A, and B is measured by a color difference meter (Σ90 color measuring system, Nippon Denshoku Inc, co., LTD, Japan), the brightness is expressed by L, and the chromaticity is A and B indicate. 13 This paper scale applies to China National Standard (CNS) A4 specification (210 X 297 mm) -I---^——l·--- (Please read the phonetic S on the back and fill in the form) Page) Ministry of Economic Affairs Intellectual Property Bureau Staff Consumer Cooperative Printed ^1 ^1 ^1 ϋ ϋ I ϋ ϋ Βϋ Βϋ ϋ ^1 ^1 ^1 ^1 ^1 ϋ ϋ III ^1 «ϋ ϋ I ϋ 1286075 A7 B7 Five , description of the invention (11) Table IV content (mg / g) VI-0 6 - ginger alcohol 17.30 ± 0.00 6 - ginger alcohol 26.29 ± 0.00 6 - dehydrogenated dione 19.20 ± 1.9 Ministry of Economic Affairs Intellectual Property Bureau employees consumption cooperatives Print Example 7: Inhibition of the inhibition of the activity of the genus Bacillus sphaeroides ((10) and oocysts) The evaluation of the inhibitory effect on the genus Bacillus sp.: Ginger extract against the genus Trichophyton 11^1^ The inhibition of &81*〇卩11>^68; octa (1^95 3 3) was evaluated according to the method of £(1'\¥&1(13,].11· et al. (Edwards, J. R· et al·, Antimicrobial Agents Chemotherapy 33: 21 5-222, 1989), and expressed in terms of minimum inhibitory concentration (MIC), the extract of the platform is dimethyl sulfoxide (DMS0). After dissolution, it is diluted to various desired concentrations of the stock solution. Add 0.01 ml of the basic solution to a 48-well plate containing 0.99 ml of potato dextrose solution (Potato Dextrose Broth (DIFCO, USA)) and 103 to 104 CFU/ml of Bacillus licheniformis at 28 °C. After 72 hours of warm bathing, the inhibition of the extract was determined by visual inspection. In the experiment, the vehicle was used as a blank control group, and each test was carried out in a double-over manner. The results are shown in Table 5. 14 This paper size is applicable to China National Standard (CNS) A4 specification (210 X 297 mm) -----^----L---- (Please read the note on the back and fill out this page) -------- 1286075 A7 __B7_____ V. Description of the invention (12) Inhibition of prion prion. The inhibitory effect of ginger extract on Pityrosporum ovale (ATCC 38593> is also based on The above methods of Edwards, JR et al. were evaluated and expressed in minimum inhibitory concentration (MIC), and the ginger extract was dissolved in DMSO and then diluted to a basic solution of the desired concentration. 0.01 ml of the basic solution was added. In a 48-well culture dish containing 0.99 ml of fluid Sacramento Media [Fluid Sabouraud Medium (DIFCO, U.S. A·)] and 1 03 -104 CFU/ml E. falciparum at 37° After 48 hours of warm bathing, the inhibitory effect of the extract was determined by visual inspection. In the experiment, solvent DMSO was used as a blank control group, and each test was carried out in a double-fold manner. The results are shown in Table 5. Table 5 Inhibition of Ginger Extract on Ophiopogonacea and Bacillus spores Ginger extract a) MIC (pg/ml) Ovipositus prion bacillus spores 1-0 100 30 I-OM 100 30 11-0 500 30 II-OC 100 30 II-OM 100 30 ΙΙΙ- 0 500 100 blank control group __ b) _ b) No inhibition in the preparation of Examples 1, 2 and 3 15 The paper size applies to the Chinese National Standard (CNS) A4 specification (210 X 297 mm) (please Read the notes on the back and fill out this page) --------Book --------- Line » Ministry of Economic Affairs Intellectual Property Bureau employee consumption cooperative printing