1262793 九、發明說明： 【發明所屬之技術領域】 本發明係有關一種草藥組合物，詳言之，係有關治療全 身性紅斑性狼瘡之組合物。 【先前技術】 全身性紅斑性狼瘡（Systemic Lupus Erythematosus，簡稱 SLE)是一種累及全身多器官、臨床表現複雜、病程長且反 覆的慢性自體免疫性疾病，其主要特徵為抗核抗體之產生 與免疫複合物之形成。其病因不明，目前認為與遺傳、病 毒或細菌感染、内分泌因素、環境因素、精神因素等諸多 因素有關。全身性紅斑性狼瘡會侵犯全身的任何器官組 織，尤其是關節、皮膚、腎臟、腦部、心肺等，其中對關 節和皮膚的影響最為顯著。此病好發於20至40歲中青年女 性，男女比值為1 ·· 8至10，發病年齡大多為15至35歲，但 亦見於兒童及老年人。其臨床症狀有：（1)特徵性皮損，如 顏面蝶形全身性紅斑，對日光和紫外線照射敏感性較高； (2)不規則不定型發熱；（3)關節疼痛，呈紅腫熱痛或多發性 關節痛；（4) 口腔枯膜全身性紅斑、糜爛、潰瘍；（5)腎臟損 害，表現腎炎或腎病症候群；（6)心臟損害，多見心包炎， 可導致心力衰竭；（7)精神、神經全身障礙；及（8)呼吸全身 損害，可發生胸膜炎、呼吸衰竭，嚴重者可危及生命 (Hochberg, M.C. ； 1997 ； Updating the American college of rheumatology revised criteria for the classification of systemic lupus erythemaosus. Arthritis & Rheum ? 40:1725-1734； Patrick M. 96177.doc 12627931262793 IX. Description of the Invention: [Technical Field of the Invention] The present invention relates to a herbal composition, in particular, to a composition for treating allergic lupus erythematosus. [Prior Art] Systemic Lupus Erythematosus (SLE) is a chronic autoimmune disease involving multiple organs, complicated clinical manifestations, long course and repeated. Its main feature is the production of anti-nuclear antibodies. The formation of immune complexes. The cause is unknown and is currently thought to be related to genetic, viral or bacterial infections, endocrine factors, environmental factors, and mental factors. Systemic lupus erythematosus invades any organ of the body, especially joints, skin, kidneys, brain, heart and lungs, etc., with the most significant effects on joints and skin. The disease occurs in young women between the ages of 20 and 40. The ratio of male to female is 1 · 8 to 10, and the age of onset is mostly 15 to 35 years old, but it is also seen in children and the elderly. The clinical symptoms are: (1) characteristic skin lesions, such as facial butterfly systemic erythema, high sensitivity to sunlight and ultraviolet radiation; (2) irregular fever; (3) joint pain, redness and heat pain Or multiple joint pain; (4) systemic erythema, erosion, ulcer in the oral mucosa; (5) kidney damage, showing nephritis or renal syndrome; (6) heart damage, more pericarditis, can lead to heart failure; ) mental and neurological systemic disorders; and (8) respiratory systemic damage, pleurisy, respiratory failure, severe life-threatening (Hochberg, MC; 1997; Updating the American college of rheumatology revised criteria for the classification of systemic lupus erythemaosus. Arthritis & Rheum ? 40:1725-1734; Patrick M. 96177.doc 1262793
Gaffney, Ward A. Ortmann, Scott A. Selby, et al. (2000) Genome screenimg in human systemic lupus erythematosus: results from a second Minnesota cohort and combined analysis of 187 sib-pire families. Am.J.Hum.Genet.66: 547-556 ； Betty P. Tsao ； 2003 ； The genetics of human systemic lupus erythematosus. TRENDS in immunology. 24(11): 595-602) o 目前對全身性紅斑性狼瘡治療有非類固醇抗炎藥 (Nonsteroidal anti-inflammatory drugs, NSAIDs);抗癔疾藥 物（anti-malarials);腎上腺皮質素（Corticosteroid，亦即類 固醇）及免疫全身抑制劑（Immunosuppressive drugs)等四類 藥物，其中主要治療藥物為腎上腺皮質素，免疫抑制劑則 用於出現蛋白尿的腎臟損傷或類固醇減量後易復發者，亦 或是二者聯合應用，然而應用此二類藥物卻會產生不可忽 視的副作用。 一直以來，中草藥被認為是副作用較少的疾病治療方 式。坊間認為某些中草藥對治療全身性紅斑性狼瘡是有幫 助的，但多屬於民間驗方或是以口授方式流傳。這些治療 的方劑通常較為複雜且由多味藥材組合而成，並未經科學 印證，至目前為止，尚未為醫學界完全掌握。因此有需要 開發簡單又有效的中草藥製劑。 目前中草藥研究最多且有效的治療全身性紅斑性狼瘡之 中草藥單味藥物為雷公藤製劑（Tripterygium wilfordii)(Jia Yongfeng，Li Hong Jianping and Zhang Luoxiu ； 2000 ； The therapeutic effect of multiglycosides of Tripterygium wilfordii 96177.doc 1262793Gaffney, Ward A. Ortmann, Scott A. Selby, et al. (2000) Genome screenimg in human systemic lupus erythematosus: results from a second Minnesota cohort and combined analysis of 187 sib-pire families. Am.J.Hum.Genet. 66: 547-556; Betty P. Tsao; 2003; The genetics of human systemic lupus erythematosus. TRENDS in immunology. 24(11): 595-602) o Currently for the treatment of systemic lupus erythematosus with non-steroidal anti-inflammatory drugs ( Nonsteroidal anti-inflammatory drugs (NSAIDs); anti-malarials; corticosteroids (also known as steroids) and immune systemic inhibitors (Immunosuppressive drugs), the main therapeutic drug is adrenal gland Quality, immunosuppressive agents are used for kidney damage caused by proteinuria or relapse after steroid reduction, or a combination of the two, but the application of these two drugs can produce non-negligible side effects. Chinese herbal medicine has long been considered as a treatment for diseases with fewer side effects. It is believed that certain Chinese herbal medicines are helpful for the treatment of systemic lupus erythematosus, but they are mostly folk prescriptions or circulated by dictation. The prescriptions for these treatments are usually complex and composed of multi-flavored herbs, which have not been scientifically proven and have not been fully understood by the medical community until now. Therefore, there is a need to develop simple and effective Chinese herbal preparations. At present, the most effective and effective treatment of systemic lupus erythematosus in Chinese herbal medicine is the Tripterygium wilfordii (Jia Yongfeng, Li Hong Jianping and Zhang Luoxiu; 2000; The therapeutic effect of multiglycosides of Tripterygium wilfordii 96177.doc 1262793
Hook f. on autoimmune syndrome induced by Campylobacter jejuniin mice. Journal of Chinese Pharmaceutical Sciences 9(3): 155-157 ;虞海燕、秦萬章、吳厚生，1999 ;雷公藤治療全 身性紅斑狼瘡免疫機制的研究；中國現代應用藥學雜誌 16(2) : 10-13 ;虞海燕、秦萬章、吳厚生，1999 ;雷公藤鹼 戊對全身性紅斑狼瘡患者B細胞免疫功能的體外實驗研 究；中國免疫學雜誌15(1) : 27-28，31 ;徐瑞安、張堂德、 詹青松，2 0 0 3 ;雷公藤内自旨醇對全身性紅斑狼瘡病人B細胞 表達CD86分子的影響；中華風濕病學雜誌7(5) ·· 275-277)， 其主要治療作用在自體免疫性疾病，而被廣泛應用於治療 風濕病和全身性紅斑性狼瘡等疾病，但雷公藤的副作為毒 性過大，易產生多全身多器官的不良反應，甚至導致死亡 (李平，陳鋼，趙怡蕊，2002 ;雷公藤的不良反應；藥物不 良反應雜誌4 : 238-240)。 職是之故，篩選一種有效及無副作用之治療藥物尤為重 要。 【發明内容】 發明概述 本發明之一目的在於提供一種治療全身性紅斑性狼瘡之 組合物，其包含以重量份數計之： 10至50%之白朮； 10至50%之杜仲； 10至50%之白花蛇舌草；及 10至50%之忍冬藤。 96177.doc 1262793 諸明之另—目的在於提供—種治療全身性紅斑性狼瘡 之卒取液，其係以包含下列步驟之方法所製備： ⑷提供以重量計之τ科藥··〗Q至卿之白朮、I技观 之杜仲、10至50%之白花蛇舌草及1〇至5〇%之忍冬藤； 及 (b)將步驟⑷之草藥在9()至12(^之溫度下分別以水萃取 後再加以混合；或將步驟⑷之草藥先行混合後在％至 120 C之溫度下以水萃取，以得到萃取液。 發明詳細說明 本發明係關於一種治療全身性紅斑性狼療之組合物，其 可有效地治療全身性紅斑性狼瘡之症狀，且無副作用。 根據本發明之治療全身性紅斑性狼瘡之組合物，其包含 以重量份數計之： 10至50%之白朮； 10至50%之杜仲； 10至5〇%之白花蛇舌草；及 10至50%之忍冬藤。 本文中所言之「全身性紅斑性狼瘡」，或稱為「系統性紅 斑性狼瘡」，亦常簡稱為「紅斑性狼瘡」，係指具有多樣臨 木症狀表現之人體自體免疫性疾病。根據本發明之組合物 可多方面緩解全身性紅斑性狼瘡之臨床症狀，而提高羅病 個體之生活品質及延長罹患全身性紅斑性狼瘡個體之壽 命。於=用根據本發明組合物治療全身性紅斑性狼瘡小鼠 之動物貫驗中，可觀察到該組合物可延長抗核抗體產生及 96177.doc 1262793 減少蛋白尿產生。 根據本發明之組合物，白朮之較佳含量為⑺至丨杜 仲之較佳含量為10至30%;白花蛇舌草之較佳含量為呢 3〇%;忍、冬藤之較佳含量為.规。於—更佳具體實施例 中，根據本發明之組合物包含以重量份數計之25%白朮、 25%杜仲、25%白花蛇舌草及25%忍冬藤。 較佳地，根據本發明之組合物另包含類固醇。類固醇係 為腎上腺分泌激素，亦為最強效之抗發炎藥劑，可在數小 時内減輕疼痛，也可控制狼瘡的症狀，因而臨床上普遍使 用於治療全身性紅斑性狼瘡。但類固醇常見的副作用包括 增重、圓臉、易齋青，高劑量類固醇亦使病人心情大變、 神經質、失眠、繁間、水份貯積、腳腫、高也壓、引發糖 尿病或惡化、增加«機會、胃出血、長期使用的患者會 有骨質疏鬆、白内障等。併用本發明之草藥與類固醇可大 幅增進治療效果並可減低類固醇之使用量。於本發明之動 物實驗中，併用草藥與類固醇於治療腎炎發病率（蛋白尿） 之效果遠較單獨使用草藥或類固醇為佳，且類固醇之使用 量較少，目而減低類固醇藥物之副作用。較佳地，該類固 醇選自強體松（prednis_)及去氫可體醇（―⑹㈣之群 組。 較佳地，根據本發明之組合物另包含免疫抑制劑。使用 免疫抑制劑治療自體免疫性疾病之主要機制在於減弱體内 細胞的免疫和發炎反應，常用於疾病的急性期，特別在於 腎臟受到侵害時。免疫抑制劑與類固醇併用時，可幫助減 96177.doc 1262793 少類固醇的劑量。併用本 g n 卓樂與免疫抑制劑可大幅 =療效果，於本發明之動物實驗 抑制劑於治療腎炎發 、兇反 遠較單獨你田p ⑶白尿）及延長存活天數之效果 制劑、J 免疫抑制劑為佳。較佳地，該免疫抑 1、自核磷酸胺（cycl〇Ph〇sphamide)。 ^發明組合物可應用為藥品、食品或膳食補充品。又， f合物可製備成各種劑型，例如，但不限於鍵劑、 膠囊、溶液或粉劑。 根據本發明之組合物可依多種方法製造。製造草藥萃取 物之傳統方法即可應用在本發明上。 本發明亦提供一種治療全身性紅斑性狼瘡之萃取液，其 係以包含下列步驟之方法所製備： 八 ⑷提供以重量計之下料藥：⑽抓之自朮、丨❻至鄉 之杜仲、10至50%之白花蛇舌草及1〇至5〇%之忍冬藤； 及 …， ⑻將步驟⑷之草藥在9G至12()。〇之温度下分別以水萃取 後再加以混合；或將步驟（a)之草藥先行混合後在9〇至 120 c之溫度下以水萃取，以得到萃取液。 較佳地，步驟（b)之水含量係為草藥之5至15倍。 本發明萃取液之製備可將白朮、杜仲、白花蛇舌草及忍 冬藤在適當溫度下分別以水萃取，再將所得萃取液混合以 獲得本發明萃取液。或者，可將白朮、杜仲、白花蛇舌草 及忍冬藤先行混合後，再於適當溫度下以水萃取該草藥混 5物而獲得本發明萃取液。 96177.doc 10 1262793 為充分萃取出草藥中之有效成分，步驟（b)中之萃取可進 行多次；亦可於進行萃取前先以水浸泡草藥。於本發明之 一具體實施例中，經初次萃取後之生藥材經過濾後，再度 以水萃取，且為使產率提高，可合併多次萃取原液，並經 濃縮取得高濃度之萃取液。較佳地，製備本發明萃取液之 方法另包含濃縮步驟之萃取液之步驟。 於本發明之一具體實施例中，該萃取液係以減壓濃縮成 粉劑，以方便使用。該減壓壓縮之方法係為本發明所屬技 術領域中具一般知識者所熟知。 根據本發明之萃取液經高效液相層析（出>1^)分析具有特 殊之指紋圖譜。例如當使用cosmosil 5C18_MS 4.6x250 mm之 管柱進行高效液相層析（HPLC)時，在254奈米波長取得 之吸收圖譜包括遲滯時間為10·263至1〇 4〇〇(D1)、1 1 257至 1 1.390(D2)、8.485(Α1)、13·972(Β1)、19·897(Β2)、24·313(Β3)、 33.617(C1)及3 6.680(C2)分鐘之峰值，其中流動相在〇至6〇分 鐘之H2〇/CH3〇H梯度，且流速為1毫升/分鐘。 茲以下列實例予以詳細說明本發明，唯並不意味本發明 僅侷限於此等實例所揭示之内容。 【實施方式】 實例一：本發明之治療全身性紅斑性狼瘡之組合物 將等重量之白朮、杜仲、白花蛇舌草及忍冬藤共丨5〇〇 g， 分別置於兩個20公升之萃取槽，並以丨5公升二次水浸泡i 小時，於約98 煎煮35分鐘後，將溶液以4〇〇號篩網過濾 後，再加入15公升水於約98 QC煎煮1小時。合併濾液以迴 96177.doc -11 - 1262793 方疋式減壓浪鈿而成乾燥粉末（代號，最終 產率為18.5%〜25%。 將萃取液以C〇sm〇sil 5C18-MS 4 6χ25〇 mm之管柱進行高 效液相層析（HPLC)。該高效液相層析之流動相在〇至6〇分 鐘之H2〇/CH3〇H梯度，且流速為i毫升/分鐘。 所得萃取液在254奈米波長之吸收圖譜之結果示於圖i， 其包括遲滯時間為10.263至1〇.4〇〇(m)、ιΐ 257至 H.390(D2) . 8.485(A1) . 13.972(B1) . 19.897(B2) ^ 24.313(B3) ^ 33.617(C1)及 36.680(C2)分鐘之峰值。 實例二··本發明t治療全身性紅斑性狼瘡之組合物對全 身性紅斑狼瘡小鼠初期抗核抗體之藥效評估 在篩述全身性紅斑狼瘡藥物的動物模式中，主要是以 NZB/WF1小鼠進行試驗。此品系小鼠之發病情形與人類全 身性紅班狼瘡患者抗核抗體、尿蛋白的產生及引發腎毒性 致死之情形有著極高之相似性。NZB/WF1小鼠雌鼠發病率 較雄氣高且嚴重，@此本試驗利用nzb/Wf_鼠篩選治療 全身性紅斑狼瘡之藥物。 、 麟與4,..使用自美國MCKSON實驗室引進八週齡大之 二ZB/WF1雌性小鼠，採自由攝食方式進食。每籠飼養*隻， 每組十二f，室溫控制於22±沈，光暗循環時間各為二、 時。小鼠於十五週齡大，開始管餵根據實例一所製備之受 試藥（DCB-SLE1)，連續十二週後，停藥並進行觀察至小= 自然死亡為止。管餵後每個月進行抗核抗體的觀察，直至 有文試小鼠死亡’即停止本測試。本試驗所有受試鼠皆沒 96177.doc 1262793 有使用安樂死方法，所有受試鼠死亡均為自然發病死亡。 其結果示於圖2，當DCB-SLE1劑量為^sg/kg/day，可明 顯抑制NZB/WF1小鼠抗核抗體之產生。 實例一 ·本發明之治療全身性紅斑性狼瘡之組合物對全 身性紅斑狼瘡小鼠尿蛋白產生之藥效評估 翁#禊4、··小鼠來源、餵養同實施例二。小鼠經管餵後， 母-週測t式蛋白尿-次，蛋白尿判定為連續兩周測定二級 以上為發病。受試鼠發病後，更改為每周測試蛋白尿一次。 其結果示於圖3,其中DCB_SLE1劑量為125g/kg/day，可 明顯抑制NZB/W F 1小鼠蛋白尿的產生。 實例四：本發明之治療全身性紅斑性狼瘡之組合物對全 身性紅斑狼瘡小鼠壽命之藥效評估 昜## 4、··小鼠來源、餵養同實施例二。小鼠於十五週Hook f. on autoimmune syndrome induced by Campylobacter jejuniin mice. Journal of Chinese Pharmaceutical Sciences 9(3): 155-157 ;虞海燕,秦万章,吴厚生, 1999; Tripterygium wilfordii treatment of systemic lupus erythematosus immune mechanism; Journal of Applied Pharmacy 16(2) : 10-13 ; Qi Haiyan, Qin Wanzhang, Wu Housheng, 1999; In vitro study of B cell immune function in patients with systemic lupus erythematosus; Chinese Journal of Immunology 15(1): 27-28, 31; Xu Ruian, Zhang Tangde, Zhan Qingsong, 2 0 0 3 ; The effect of tripterygium wilfordii on the expression of CD86 molecules in B cells of patients with systemic lupus erythematosus; Chinese Journal of Rheumatology 7(5) ·· 275 -277), its main therapeutic effect is in autoimmune diseases, and is widely used in the treatment of diseases such as rheumatism and systemic lupus erythematosus, but the side of Tripterygium wilfordii is too toxic, and it is easy to produce multiple systemic and multiple organ adverse reactions. And even led to death (Li Ping, Chen Gang, Zhao Yirui, 2002; adverse reactions of Tripterygium wilfordii; Journal of Adverse Drug Reactions 4: 238-240). For this reason, it is especially important to screen a therapeutic drug that is effective and has no side effects. SUMMARY OF THE INVENTION One object of the present invention is to provide a composition for treating systemic lupus erythematosus comprising, by weight: 10 to 50% of atractylodes; 10 to 50% of Eucommia; 10 to 50 % of Hedyotis diffusa; and 10 to 50% of honeysuckle. 96177.doc 1262793 The other purpose of Zhuming is to provide a treatment solution for systemic lupus erythematosus, which is prepared by the method comprising the following steps: (4) Providing τ medicines by weight··〗 Q Zhiqing Atractylodes Rhizome, I technological eucommia, 10 to 50% of Hedyotis diffusa, and 1 to 5 % of Lonicera japonica; and (b) Step (4) of the herb at 9 () to 12 (^ temperature respectively After extracting with water, mixing; or mixing the herbs of step (4) and extracting with water at a temperature of from % to 120 C to obtain an extract. DETAILED DESCRIPTION OF THE INVENTION The present invention relates to a treatment for systemic erythematous wolf a composition for effectively treating the symptoms of systemic lupus erythematosus without side effects. The composition for treating systemic lupus erythematosus according to the present invention comprises, by weight: 10 to 50% of atractylodes; 10 to 50% eucommia; 10 to 5% of Hedyotis diffusa; and 10 to 50% of honeysuckle vine. "Systemic lupus erythematosus", or "systemic lupus erythematosus" , also often referred to as "erythema lupus", refers to a variety of A human autoimmune disease manifested by wood symptoms. The composition according to the present invention can alleviate the clinical symptoms of systemic lupus erythematosus in a multi-faceted manner, and improve the quality of life of an individual with Luo disease and prolong the life of an individual with systemic lupus erythematosus. In the examination of animals treated with the composition according to the invention for mice with systemic lupus erythematosus, it was observed that the composition prolongs anti-nuclear antibody production and 96177.doc 1262793 reduces proteinuria production. Composition according to the invention The preferred content of Atractylodes is (7) to 10~30% of the content of Eucommia ulmoides; the preferred content of Hedyotis diffusa is 3〇%; the preferred content of tolerance and winter vines is . In a preferred embodiment, the composition according to the invention comprises 25% atractylodes, 25% Eucommia, 25% Hedyotis diffusa and 25% Lonicera japonica in parts by weight. Preferably, the composition according to the invention additionally Contains steroids. Steroids are adrenal hormones, also the most potent anti-inflammatory agent, can relieve pain within a few hours, can also control the symptoms of lupus, so it is commonly used in the treatment of systemic erythema Lupus. But the common side effects of steroids include weight gain, round face, easy to eat, high doses of steroids also make the patient's mood change, nervousness, insomnia, interstitial, water storage, swollen feet, high pressure, trigger diabetes or Deterioration, increase of "opportunity, stomach bleeding, long-term use of patients with osteoporosis, cataracts, etc.. And the use of the herbs and steroids of the present invention can greatly improve the therapeutic effect and reduce the amount of steroids. In the animal experiment of the present invention, use together The effect of herbs and steroids in the treatment of nephritis (proteinuria) is much better than the use of herbs or steroids alone, and the use of steroids is less, thus reducing the side effects of steroids. Preferably, the steroid is selected from the group consisting of prednisone. (prednis_) and dehydrocoholic alcohol (-(6) (d) group. Preferably, the composition according to the invention further comprises an immunosuppressive agent. The main mechanism for the use of immunosuppressive agents for the treatment of autoimmune diseases is to attenuate the immune and inflammatory responses of cells in the body, often in the acute phase of the disease, especially when the kidneys are infested. When used together with steroids, immunosuppressive agents can help reduce the dose of steroids. And the use of the gn Zhuo Le and the immunosuppressive agent can greatly reduce the therapeutic effect, in the animal experimental inhibitor of the present invention, in the treatment of nephritis, the fierce anti-far more than the individual p (3) white urine) and the prolonged survival effect of the preparation, J immunization Inhibitors are preferred. Preferably, the immunosuppressive hormone is cycl〇Ph〇sphamide. The inventive composition can be applied as a pharmaceutical, food or dietary supplement. Further, the f compound can be prepared in various dosage forms such as, but not limited to, a key, a capsule, a solution or a powder. The compositions according to the invention can be made in a variety of ways. A conventional method of producing an herbal extract can be applied to the present invention. The invention also provides an extract for treating systemic lupus erythematosus, which is prepared by the method comprising the following steps: (8) providing the medicine under the weight: (10) grasping the self-surgery, sputum to the town eucommia, 10 to 50% of Hedyotis diffusa and 1〇 to 5〇% of Lonicera japonica; and..., (8) The herb of step (4) is 9G to 12(). The mixture is extracted with water and mixed with water; or the herb of step (a) is first mixed and extracted with water at a temperature of 9 to 120 c to obtain an extract. Preferably, the water content of step (b) is 5 to 15 times that of the herbal medicine. The extract of the present invention can be prepared by extracting Atractylodes macrocephala, Eucommia ulmoides, Hedyotis diffusa and Lonicera edulis respectively at appropriate temperature with water, and then mixing the obtained extracts to obtain the extract of the present invention. Alternatively, the extract of the present invention can be obtained by first mixing Atractylodes macrocephala, Eucommia ulmoides, Hedyotis diffusa and Lonicera edulis, and then extracting the herbal mixture with water at an appropriate temperature. 96177.doc 10 1262793 In order to fully extract the active ingredients in the herbs, the extraction in step (b) can be carried out several times; the herbs can also be soaked in water before the extraction. In a specific embodiment of the present invention, the raw medicinal material after the initial extraction is filtered and then extracted with water, and in order to increase the yield, the extract may be combined for multiple times, and concentrated to obtain a high concentration of the extract. Preferably, the method of preparing the extract of the present invention further comprises the step of concentrating the extract. In one embodiment of the invention, the extract is concentrated under reduced pressure to a powder for ease of use. The method of decompression compression is well known to those of ordinary skill in the art to which the invention pertains. The extract according to the present invention has a special fingerprint by high performance liquid chromatography (out > 1^) analysis. For example, when performing high performance liquid chromatography (HPLC) on a cosmosil 5C18_MS 4.6x250 mm column, the absorption spectrum obtained at a wavelength of 254 nm includes a hysteresis time of 10.263 to 1〇4〇〇(D1), 1 1 257 to 1 1.390 (D2), 8.485 (Α1), 13·972 (Β1), 19·897 (Β2), 24·313 (Β3), 33.617 (C1) and 3 6.680 (C2) minutes peak, where flow The gradient was H2〇/CH3〇H in 〇 to 6 〇 minutes, and the flow rate was 1 ml/min. The invention is illustrated by the following examples, which are not intended to be construed as limiting the invention. [Examples] Example 1: The composition for treating systemic lupus erythematosus of the present invention comprises equal weight of Atractylodes Rhizome, Eucommia, Hedyotis diffusa, and Lonicera japonica 5 〇〇g, respectively, in two 20 liter extracts The tank was immersed in 丨5 liters of secondary water for 1 hour, and after boiling for about 30 minutes at about 98, the solution was filtered through a 4 筛 sieve, and then 15 liters of water was added to decoct at about 98 QC for 1 hour. The combined filtrates were returned to a dry powder of 96177.doc -11 - 1262793 square decompression to obtain a dry powder (code, the final yield was 18.5%~25%. The extract was C〇sm〇sil 5C18-MS 4 6χ25〇 The column of mm was subjected to high performance liquid chromatography (HPLC). The mobile phase of the high performance liquid chromatography was subjected to a H2〇/CH3〇H gradient of 〇 to 6 〇 minutes, and the flow rate was i ml/min. The results of the absorption spectrum at 254 nm wavelength are shown in Figure i, which includes a lag time of 10.263 to 1 〇.4 〇〇 (m), ι 257 to H.390 (D2). 8.485 (A1) . 13.972 (B1) 19.897(B2) ^ 24.313(B3) ^ 33.617(C1) and 36.680(C2) minutes peak. Example 2 · The composition of the present invention for treating systemic lupus erythematosus to the initial antinuclear of mice with systemic lupus erythematosus Efficacy evaluation of antibodies In the animal model of screening for systemic lupus erythematosus drugs, it is mainly tested in NZB/WF1 mice. The incidence of this strain is the anti-nuclear antibody and urine protein of human systemic lupus erythematosus patients. There is a very high similarity between the occurrence and the cause of nephrotoxicity. The incidence of female NZB/WF1 mice is higher than that of males. Seriously, @this test uses nzb/Wf_ mice to screen for the treatment of systemic lupus erythematosus., Lin and 4,.. Using the American MCKSON laboratory to introduce eight-week-old two ZB/WF1 female mice, free Ingestion mode to eat. Each cage is kept only *, each group is 12 f, the room temperature is controlled at 22 ± Shen, and the light and dark cycle time is 2 hours. The mice are 15 weeks old and start feeding according to the example one. The prepared test drug (DCB-SLE1) was stopped for 12 weeks and observed until it was small = natural death. After the tube was fed, the antinuclear antibody was observed every month until the test mice died. That is, the test was stopped. All the rats in the test did not use 96177.doc 1262793. The euthanasia method was used, and all the rats died were natural morbid death. The results are shown in Fig. 2, when the dose of DCB-SLE1 is ^sg/kg. /day, can significantly inhibit the production of anti-nuclear antibodies in NZB/WF1 mice.Example I. Evaluation of the efficacy of the composition of the present invention for treating systemic lupus erythematosus on urine protein production in mice with systemic lupus erythematosus Weng#禊4 Mouse source and feeding are the same as in Example 2. After the mice are fed by the tube Maternal-weekly test for t-type proteinuria-time, proteinuria was determined to be more than two consecutive weeks for the onset of disease. After the onset of the test mice, the protein urine was changed once a week. The results are shown in Figure 3, in which DCB_SLE1 dose was administered. 125g/kg/day can significantly inhibit the production of proteinuria in NZB/WF 1 mice. Example 4: The efficacy of the composition of the present invention for treating systemic lupus erythematosus on the lifespan of mice with systemic lupus erythematosus 昜# #4,·· The source of the mouse and feeding were the same as in the second example. Mouse in fifteen weeks
齡大，開始f餃受試藥，連續十二週後，停藥並進行觀察 至小鼠自然死亡為止。 T 其結果示於圖4,其中DCB_SLE1劑量為125g/kg/day，可 明顯延長NZB/WF1小鼠之壽命。 實例五：本發明之併用草藥及免疫抑制劑之組合物對全 身性紅斑狼瘡小鼠蛋白尿產生之藥效評估 始影、鼠㈣、餵養同實施週齡開 二踏母天管餵一次及每10天腹腔注射環磷酸胺（CTX)或生理 艮鹽水一次，連續6週，停藥2週，視為一個療程，丘 管㈣每二週測試蛋白尿—次，*白尿判定為連 Λ。測定二級以上為發病。受試氣發病後，更改為每周 96177.doc -13- 1262793 測試蛋白尿一次。 其結果示於圖5，其中1.25 g/kg/day之DCB-SLE1結合16.7 mg/kg/1 Oday之CTX之治療可明顯抑制蛋白尿之產生。 實例六：本發明之併用草藥及免疫抑制劑之組合物對全 身性紅斑狼瘡小鼠壽命之藥效評估 動物模式：小鼠來源、餵養同實施例二。小鼠1 5週齡開 始，每天管餵一次及每10天腹腔注射CTX或生理食鹽水一 次，連續6週，停藥2週，視為一個療程。進行3個療程後， 停藥並進行觀察至小鼠自然死亡為止。小鼠依給藥共分四 組： A ： DCB-SLE1 1.25 g/kg/day B ： DCB-SLE1 12.5 g/kg/day C ： DCB-SLE1 1.25 g/kg/day+ CTX16.5 mg/kg/10 day D ： CTX16.5 mg/kg/lOday 其結果示於圖6，其中1.25g/kg/day之DCB-SLE1結合 16.7mg/kg/10day之CTX之治療可明顯延長NZB/W FI小鼠 壽命。 實例七：本發明之併用草藥及類固醇之組合物對全身性 紅斑狼瘡小鼠腎炎發病率之藥效評估 動物模式：小鼠來源、餵養同實施例二，小鼠15週齡開 始，每天管餵一次，連續6週，停藥2週，視為一個療程， 共進行3個療程。觀察腎炎發病率至對照組小鼠全部發病為 止。小鼠依給藥方式分成七組： A ： DCB-SLE1 1.25 mg/kg/day 96177.doc -14- 1262793 B : DCB-SLEl 1.25 mg/kg/day+去氫可體醇0·5 mg/kg/day C: DCB-SLE1 1.25mg/kg/day+去氫可體醇2mg/kg/day D : DCB-SLE1 12.5 mg/kg/day+去氫可體醇 0.5 mg/kg/day E: DCB-SLE1 12.5mg/kg/day+去氫可體醇2mg/kg/day F :去氫可體醇0.5 mg/kg/day G :去氫可體醇2 mg/kg/day 其結果示於圖7，當對照組腎炎發病率為100%時 DCB-SLE1結合類固醇組（B、C、D、E)的發病率最低，其 中又以C、D兩組最好。1.25 g/kg/day之DCB-SLE1結合2 mg/kg/day之去氳可體醇之治療可明顯抑制NZB/W F1小鼠 之蛋白尿產生。 實例八：本發明併用草藥及類固醇組合物對全身性紅斑 狼瘡小鼠壽命之藥效評估 參#禊式··小鼠來源、餵養同實施例二。小鼠1 5週齡開 始，每天管餵一次，連續6週，停藥2週，視為一個療程， 共進行3個療程。壽命之藥效評估係進行觀察至對照組小鼠 死亡率為百分之五十為止。 其結果示於表1 : 表1 : 組 別 50%小鼠死亡（天） 對照組 262.6 ± 46.7 DCB-SLE1 1.25 mg/kg/day 334.0 ±40.6* DCB-SLE1 1.25 mg/kg/day+去氫可 體醇 0.5 mg/kg/day 356.2 ± 19.5" DCB-SLE1 1.25 mg/kg/day+去氫可 體醇 2 mg/kg/day 416.6 ± 49.7" * DCB-SLE1 12.5 mg/kg/day+去氫可 體醇 0.5 mg/kg/day 379.8 ± 25.5* * * 96177.doc -15- 1262793 DCB-SLE1 12.5 mg/kg/day+去氫可 體醇 2 mg/kg/day 368.4 ± 27.2* * * 去氫可體醇〇·5 mg/kg/day 281.1 ± 56.2 去氳可體醇2 mg/kg/day 338.8 ± 54.Γ 實驗組與對照組比，*Ρ<〇·〇1、* *Ρ<〇·〇(Π、* * *ρ<0·0001 由表1可知，結合12.5g/kg/day之DCB-SLE1結合 2mg/kg/day之去氫可體醇之治療可明顯延長NZB/W F 1小鼠 壽命。 上述實施例僅為說明本發明之原理及其功效，而非限制 本發明。習於此技術之人士對上述實施例所做之修改及變 化仍不違背本發明之精神。本發明之權利範圍應如後述之 申請專利範圍所列。 【圖式簡單說明】 圖1為根據本發明之卒取液之南效液相層析結果圖。When the age is large, the test is started. After 12 weeks, the drug is stopped and observed until the mouse naturally dies. The results of T are shown in Figure 4, where the DCB_SLE1 dose was 125 g/kg/day, which significantly prolonged the lifespan of NZB/WF1 mice. Example 5: The efficacy evaluation of the combination of the herbal medicine and the immunosuppressant of the present invention on the proteinuria of the systemic lupus erythematosus mouse, the mouse (4), the feeding and the implementation of the weekly age, the second step of the mother tube feeding once and each 10 days of intraperitoneal injection of cyclophosphamide (CTX) or physiological sputum saline once for 6 weeks, discontinued for 2 weeks, regarded as a course of treatment, the mound tube (four) every two weeks to test proteinuria - times, * white urine was determined as flail. The above two levels are measured as the onset. After the onset of test gas, change to 96177.doc -13 - 1262793 per week to test proteinuria once. The results are shown in Figure 5, in which treatment with 1.25 g/kg/day of DCB-SLE1 in combination with 16.7 mg/kg/1 Oday of CTX significantly inhibited proteinuria production. Example 6: Efficacy evaluation of the combination of the herbal medicine and the immunosuppressant of the present invention on the lifespan of the whole body lupus erythematosus. Animal model: the source of the mouse and the feeding were the same as in the second embodiment. The mice were started at 5 weeks of age, once a day, and once every 10 days, intraperitoneally injected with CTX or saline once every 6 weeks for 2 weeks, which was regarded as a course of treatment. After 3 courses of treatment, the drug was stopped and observed until the mice died naturally. Mice were divided into four groups according to their administration: A : DCB-SLE1 1.25 g/kg/day B : DCB-SLE1 12.5 g/kg/day C : DCB-SLE1 1.25 g/kg/day+ CTX16.5 mg/kg/ 10 day D : CTX16.5 mg/kg/lOday The results are shown in Figure 6. The treatment of 1.25g/kg/day DCB-SLE1 combined with 16.7mg/kg/10day CTX can significantly prolong NZB/W FI mice. life. Example 7: Efficacy evaluation of the combination of herbal medicine and steroid of the present invention on the incidence of nephritis in mice with systemic lupus erythematosus Animal model: mouse source, feeding as in the second embodiment, the mouse starts at 15 weeks of age, and the tube is fed every day. Once, for 6 weeks, 2 weeks of withdrawal, as a course of treatment, a total of 3 courses. The incidence of nephritis was observed to the extent of all the mice in the control group. Mice were divided into seven groups according to the mode of administration: A : DCB-SLE1 1.25 mg/kg/day 96177.doc -14- 1262793 B : DCB-SLEl 1.25 mg/kg/day + dehydrocohol alcohol 0·5 mg/kg /day C: DCB-SLE1 1.25mg/kg/day+dehydrocohol 2mg/kg/day D : DCB-SLE1 12.5 mg/kg/day+dehydrocohol 0.5mg/kg/day E: DCB-SLE1 12.5 mg/kg/day + dehydrocohol alcohol 2 mg/kg/day F: dehydrocohol alcohol 0.5 mg/kg/day G: dehydrocohol alcohol 2 mg/kg/day The results are shown in Figure 7, when When the incidence of nephritis was 100% in the control group, the incidence of DCB-SLE1 combined with steroids (B, C, D, E) was the lowest, and the best was in groups C and D. Treatment with 1.25 g/kg/day of DCB-SLE1 in combination with 2 mg/kg/day of decyl alcohol significantly inhibited proteinuria production in NZB/W F1 mice. Example 8: Evaluation of the efficacy of the herb and steroid composition of the present invention on the lifespan of mice with systemic lupus erythematosus. The source of the mouse was fed and the same as in the second example. The mice were started at 5 weeks of age, once a day for 6 weeks, and stopped for 2 weeks. It was regarded as a course of treatment for a total of 3 courses. The efficacy evaluation of the lifespan was observed until the mortality of the control mice was 50%. The results are shown in Table 1: Table 1: Group 50% mouse death (days) Control group 262.6 ± 46.7 DCB-SLE1 1.25 mg/kg/day 334.0 ± 40.6* DCB-SLE1 1.25 mg/kg/day+ dehydrogenation Body alcohol 0.5 mg/kg/day 356.2 ± 19.5" DCB-SLE1 1.25 mg/kg/day+ dehydrocohol 2 mg/kg/day 416.6 ± 49.7" * DCB-SLE1 12.5 mg/kg/day+ dehydrogenation Sterol 0.5 mg/kg/day 379.8 ± 25.5* * * 96177.doc -15- 1262793 DCB-SLE1 12.5 mg/kg/day + dehydrocohol 2 mg/kg/day 368.4 ± 27.2* * * Dehydrogenation Sterol 〇·5 mg/kg/day 281.1 ± 56.2 steroidal alcohol 2 mg/kg/day 338.8 ± 54.Γ Experimental group compared with control group, *Ρ<〇·〇1,* *Ρ<〇 ·〇(Π,* * *ρ<0·0001 As can be seen from Table 1, treatment with 12.5g/kg/day of DCB-SLE1 combined with 2mg/kg/day of dehydrocohol can significantly extend NZB/WF 1 The present invention is not intended to limit the scope of the present invention. The present invention is not intended to limit the scope of the present invention. The scope of rights should be as follows BRIEF DESCRIPTION OF THE DRAWINGS [Fig. 1] Fig. 1 is a graph showing the results of a southern liquid chromatography of a stroke liquid according to the present invention.
圖2為DCB-SLE1對NZB/W F1小鼠血清中抗ds-DNA IgG 抗體的效應圖。 圖3為DCB-SLE1對NZB/W F1小鼠蛋白尿的影響圖。 圖4為DCB-SLE1對NZB/W F1小鼠壽命的影響圖，其中實 驗組與對照組比，*P<0.01。 圖5為DCB-SLE1與CTX結合使用對NZB/W F1小鼠蛋白 尿的影響圖。 圖6為DCB-SLE1與CTX結合使用對NZB/W F1小鼠壽命 的影響圖，其中實驗組與對照組比，*ρ<〇·〇Γ *ρ<〇.〇〇1。 圖7為DCB-SLE1結合去氫可體醇對NZB/W F1小鼠腎炎 發病率（％)的效應圖，其中實驗組與對照組比，*Ρ<〇·〇1。 96177.doc 16Figure 2 is a graph showing the effect of DCB-SLE1 on anti-ds-DNA IgG antibodies in NZB/W F1 mouse serum. Figure 3 is a graph showing the effect of DCB-SLE1 on proteinuria in NZB/W F1 mice. Figure 4 is a graph showing the effect of DCB-SLE1 on the lifespan of NZB/W F1 mice, where the experimental group was compared with the control group, *P < 0.01. Figure 5 is a graph showing the effect of DCB-SLE1 in combination with CTX on proteinuria in NZB/W F1 mice. Figure 6 is a graph showing the effect of DCB-SLE1 in combination with CTX on the lifespan of NZB/W F1 mice, where the experimental group is compared with the control group, *ρ<〇·〇Γ *ρ<〇.〇〇1. Fig. 7 is a graph showing the effect of DCB-SLE1 combined with dehydrocoholitol on the incidence (%) of nephritis in NZB/W F1 mice, wherein the experimental group is compared with the control group, *Ρ<〇·〇1. 96177.doc 16