13114料牛 ?:第 91118900號專利申請案 中文說明書替換Μ 民έΐ 93年9月23日修正 五、發明説明（1) 本發明係關於用於預防及/或治療中風與絕血狀況之醫 藥組成物，其中高密度脂蛋白粒子，較宜是再構建高密度 脂蛋白（rHDL)粒子是用藥至對其有需要之病人，特別是經 由靜脈輸注。 中風可以分成血栓栓塞性及出血性形式且在西方國家 是第三大的死亡原因，列在心臟病及癌症之後，在美國每 年有600000人罹患新的或復發的中風（約500000人是第一 次發作）且其中約29%在第一年內死亡（1)，中風之發病率隨 著年齡增加，且是美國的老年人中嚴重長期病殘之首要原 因，總成本是513億美元/年（1)，雖然中風之死亡率在近年 來已經降低，大部分是因爲增加警覺性及風險因子例如高 血壓、高膽固醇、心律不整或糖尿病之較佳控制。因爲老 年人口增加，中風死亡之實際數目是增加，但是當預防測 量失敗時，只有有限且有風險的溶血栓劑途徑存在，例如 t-PA (組織纖溶酶原激活物）。神經元的保護在未來可能變 成中風治療之新穎且安全的策略（2-4)。 循環休克的一個常見原因是與創傷有關之嚴重血液流失，雖 然強化監護藥劑之改進，出血性休克之死亡仍然很高（5,6)， 因此，仍然非常需求新的方向以改進出血性休克病人之治療 及演變（6)，在臨床實務中，出血性休克導致延遲的血管代償 能力消失（起因於嚴重的低血壓），且約25%的病人有數種器 官包括肺、腎、腸、肝及腦之功能障礙或衰竭（7)，另外，也 可從絕血事件發生器官功能障礙，明顯不同於出血之狀況， 該絕血事件因堵塞結果而減少血液供應，也有證據顯示再灌 本紙張尺度適用中國國家標準（CNS ) A4規格（210X297公釐） t衣-- (請先閲讀背面之注意事項再填寫本頁) 訂 線 經濟部智达財產局員工消費合作社印製 -4- 1311485 經濟部智慧財產局8工消費合作社印製 A7 B7五、發明説明（2 ) 注（復甦期間）與多種器官功能障礙徵候群（MODS)之病理有 關（8)。 根據W0 01/13939及（21),在大鼠出血性休克模式中使 用rHDL證明可大幅減少器官傷害，出血性休克包括普遍減 少血液供應至整個身體，其造成低氧傷害而影響全部器官 及組織，相反地，絕血是描述局部停止血液供應至特定器 官及組織，導致這些受影響的區域快速發生缺氧，因此傷 害之機制相當獨特。 rHDL經顯示可刺激膽固醇從末梢細胞排出，此過程較 宜稱爲逆向膽固醇運輸，而且，rHDL劑量相關地結合細菌 脂多糖（LPS)及抑制LPS引發的細胞活素製造以及PMNs (多 形核白血球）附著至內皮細胞（21)，rHDL具有抗發炎及自由 態氧游離基淸除劑活性，rHDL也降低血小半具及的速率及 程度，最近經由前臂體積掃描術測定，證明rHDL可快速恢 復內皮功能且因而在尚膽固醇症病人中經由增加氧化氮生 物利用度使血液流動正常化（9)。 中風病理之特徵是廣泛的穩態、血液流動及代謝失常 例如血栓形成、受損的內皮功能及活化的發炎級聯，也就 是增加細胞活素製造及表達附著分子（1 0-1 5),中風之另一 個特徵是在再灌流後增加的氧化應力，其咸信是在病症發 展中扮演有害的角色。 長期絕血導致增加細胞內Ca + +且因而發生的蛋白酶及 磷脂酶之活化導致形成許多膜脂質分解潛在危害的產物， 這些包括花生四烯酸代謝物，其在再灌流期間在氧氣存在 本紙張尺度適用中國國家標準（CNS ) A4規格（210X297公釐） (請先閲讀背面之注意事項再填寫本頁) 1311485 A7 B7 五、發明説明（3) (請先聞讀背面之注意事項再填寫本頁) 下，提供游離基形成之來源（例如超氧化物及羥基陰離子）， 這些游離基引發血腦障蔽破壞及神經元脫噬作用及/或壞 死，脫噬作用是一種細胞死亡形式其在無發炎回應下消除 妥協或多餘的細胞且有許多變形及生化特徵不同於壞死， 脫噬作用之特性可以在絕血傷害後見於神經及元及神經膠 質，不是暴露在致命絕血之絕血紐巴（pneumbra)之神經元可 能進行延遲的脫噬作用（1 6)，所稱的紐巴是一個腦部區域其 中血液流動降低至一個程度其中斷神經元功能及因而發生 的電子活動，但容許維持膜抽動及保持離子梯度，此腦區 域有兩個特徵可解釋其潛在的臨床重要性：1)臨床及電子 功能之中斷在此區域之特徵是基本地可逆，但2)可逆性有 時間限制且牽涉至再灌流。 經濟部智慧財產局員工消費合作社印製 訝異地，發現損傷大小在用於中風的動物模式中（外細 胞毒性及腦動脈閉合）經由用藥HDL而降低，這些數據顯示 HDL可改善外細胞毒性及絕血/再灌流神經元傷害之後果， 特別是在絕血區域及紐巴之脫噬作用及/或壞死，而且，在 出血性休克之動物模式中，其顯示HDL降低PMN浸入及防 止器官傷害及供障礙，目前，作用之機制不明，雖然不希 望局限於理論，HDL可能作爲自由態氧游離基淸除劑、血 管擴張劑，例如經由改善NO生物利用度導致改進側枝血液 流動或展現抗發炎效應，因此，HDL可作爲神經保護劑特 別是在腦血管疾病，其也可能經由結合全部這些活性而作 用，達到尙未見於目前醫療之床效應。 本發明廣泛地關於HDL用於預防及/或治療絕血或再灌 本紙張尺度適用中國國家標準（CNS ) A4規格( 210X297公釐） "' 1311485 A7 B7 五、發明説明（4) 注傷害之用途，器官絕血是因爲血液供應中斷而發生，且 在其最廣義下可導致器官供障礙或傷害，尤其是心臟、 腦、腎、肝或肺，其是局部事件/中斷而導致完全或部份且 在某些情形下可逆的傷害，發生再灌流傷害是因爲氧化的 血液快速回到絕血後的區域之結果且通常稱爲心血管及腦 過失。 因此，本發明之一個主題是使用HDL製造藥劑用於預 防及/或治療絕血或再灌注傷害之用途，具體地說，HDL可 用於預防及/或治療選自絕血中風、絕血組織傷害例如器官 之絕血傷害、心臟絕血、心臟再灌流傷害及器官移植產生 的倂發症例如腎、心臟及肝臟或心-肺導管手術之病症及其 他病症，更訝異地，經發現HDL在當暫時或永久閉合發生 時具有有利的效應，因此，造成閉合之血凝塊或其他實體 被溶解或去除對於功效並不是先決條件，而且，在絕血事 件後6或更多小時用藥HDL顯示仍然有用處，另一個訝異 的觀察是在絕血事件前用藥HDL具有有利的效應。 本發明之其他具體實施例是關於HDL用於預防及/或治 療臨時絕血發作（TIA)之用途，TIAs是常見且約三分之一將 在未來發展中風，TIA之最常見原因是從大血管（通常是狹 窄的動脈粥樣頸動脈）中的動脈粥樣硬化斑片形成血栓，因 爲HDL具有抗動脈粥樣硬化的性質，如同經由氧化氮的生 物利用度之共鳴探討內皮功能之硏究顯示，在血管張力及 結構之調整（9)中，咸信HDL可以在造成TIAs之動脈粥樣 硬化斑片安定話，因而降低主要中風之風險，目前用於 本紙張尺度適用中國國家標準（CNS ) Α4規格（210X297公釐） (請先閲讀背面之注意事項再填寫本頁) I·裝.13114 牛牛?: Chinese Patent Specification No. 91118900 Replacement Μ Minsheng September 23, 1993 Amendment 5, Invention Description (1) The present invention relates to a pharmaceutical composition for preventing and/or treating stroke and blood loss Preferably, the high density lipoprotein particles, preferably reconstituted high density lipoprotein (rHDL) particles, are administered to a patient in need thereof, particularly via intravenous infusion. Stroke can be divided into thromboembolic and hemorrhagic forms and is the third leading cause of death in Western countries. After heart disease and cancer, 600,000 people in the United States suffer from new or recurrent strokes each year (about 500,000 people are the first). Secondary seizures) and about 29% of them die within the first year (1), the incidence of stroke increases with age, and is the leading cause of severe long-term disability in the elderly in the United States, with a total cost of $51.3 billion per year. (1) Although the mortality rate of stroke has decreased in recent years, most of it is due to increased alertness and risk factors such as hypertension, high cholesterol, arrhythmia or diabetes. Because of the increase in the elderly population, the actual number of stroke deaths is increased, but when preventive measures fail, there is only a limited and risky thrombolytic route, such as t-PA (tissue plasminogen activator). The protection of neurons may become a novel and safe strategy for stroke treatment in the future (2-4). A common cause of circulatory shock is severe blood loss associated with trauma. Although the improvement of intensive care agents, the death of hemorrhagic shock remains high (5,6), so new directions are still needed to improve patients with hemorrhagic shock. Treatment and evolution (6), in clinical practice, hemorrhagic shock leads to delayed vascular compensatory capacity (caused by severe hypotension), and about 25% of patients have several organs including lung, kidney, intestine, liver and Dysfunction or failure of the brain (7). In addition, organ dysfunction can also occur from an anesthetized event, which is significantly different from the condition of bleeding. The blood-suppressed event reduces blood supply due to clogging. There is also evidence that the paper size is refilled. Applicable to China National Standard (CNS) A4 Specification (210X297 mm) t--- (Please read the notes on the back and fill out this page) Customs Ministry of Economic Affairs Zhida Property Bureau Staff Consumer Cooperative Printed -4- 1311485 Ministry of Economic Affairs Intellectual Property Bureau 8 Workers Consumption Cooperative Printed A7 B7 V. Invention Description (2) Note (during recovery) and pathology of multiple organ dysfunction syndrome (MODS) Off (8). According to W0 01/13939 and (21), the use of rHDL in rat hemorrhagic shock mode has been shown to significantly reduce organ damage. Hemorrhagic shock includes a general reduction in blood supply to the entire body, which causes hypoxia damage and affects all organs and tissues. Conversely, hematopoiesis is a description of the local cessation of blood supply to specific organs and tissues, resulting in rapid hypoxia in these affected areas, so the mechanism of injury is quite unique. rHDL has been shown to stimulate cholesterol excretion from peripheral cells, a process known as reverse cholesterol transport, and rHDL dose-associated with bacterial lipopolysaccharide (LPS) and inhibition of LPS-induced cytokine production and PMNs (polymorphonuclear leukocytes) Attached to endothelial cells (21), rHDL has anti-inflammatory and free-state oxygen free scavenger activity, and rHDL also reduces the rate and extent of hemorrhage. Recently, it was confirmed by forearm volume scanning that rHDL can quickly restore endothelial cells. Function and thus normalization of blood flow by increasing nitric oxide bioavailability in patients with cholesterol (9). Stroke pathology is characterized by extensive homeostasis, blood flow and metabolic disorders such as thrombosis, impaired endothelial function, and activated inflammatory cascades, that is, increased cytokine production and expression of attached molecules (1 0-1 5), Another feature of stroke is the increased oxidative stress after reperfusion, which is a detrimental role in the development of the condition. Long-term anemia causes increased intracellular Ca + + and the activation of proteases and phospholipases that result in the formation of many potential products of membrane lipid breakdown, including arachidonic acid metabolites, which are present in oxygen during reperfusion The scale applies to the Chinese National Standard (CNS) A4 specification (210X297 mm) (please read the notes on the back and fill out this page) 1311485 A7 B7 V. Invention description (3) (Please read the notes on the back and fill in the form) Under the page, provide sources of free radical formation (such as superoxides and hydroxyl anions), which cause blood-brain barrier damage and neuronal phagocytosis and/or necrosis, which is a form of cell death Inflammatory response eliminates compromise or excess cells and has many deformations and biochemical features different from necrosis. The characteristics of phagocytosis can be seen in nerves and neurons and glia after hemorrhagic injury, not in the blood of the deadly blood. Neurons in (pneumbra) may undergo delayed dephasing (1 6), a so-called neoba is a brain region where blood flow is reduced To the extent that it interrupts neuronal function and the resulting electronic activity, but allows membrane twitching and maintains ion gradients, this brain region has two characteristics that explain its potential clinical importance: 1) disruption of clinical and electronic functions The characteristics of the area are basically reversible, but 2) the reversibility is time-limited and involves reperfusion. The Ministry of Economic Affairs’ Intellectual Property Office staff consumption cooperative printed and surprised that the size of the lesion was reduced in the animal model for stroke (external cytotoxicity and cerebral arterial occlusion) via HDL. These data show that HDL can improve external cytotoxicity and After the injury of the blood-supplemented/reperfused neurons, especially in the hematopoietic region and neonatal phagocytosis and/or necrosis, and in the animal model of hemorrhagic shock, it shows that HDL reduces PMN immersion and prevents organ damage. And for the obstacles, at present, the mechanism of action is unknown. Although it is not desirable to be limited to theory, HDL may act as a free oxygen free radical scavenger, a vasodilator, for example, by improving NO bioavailability, resulting in improved collateral blood flow or anti-inflammatory. Effect, therefore, HDL can act as a neuroprotective agent, particularly in cerebrovascular diseases, and it may also act by combining all of these activities to achieve a medical bed effect. The present invention relates widely to the use of HDL for the prevention and/or treatment of blood-suppressed or re-perfused paper. The Chinese National Standard (CNS) A4 specification (210X297 mm) "' 1311485 A7 B7 5. Inventive Note (4) Injection injury Its use, organ arrhythmia occurs due to interruption of blood supply, and in its broadest sense can cause organs to provide obstacles or injuries, especially the heart, brain, kidney, liver or lungs, which are local events/interruptions leading to complete or Partial and in some cases reversible damage, reperfusion injury occurs as a result of oxidized blood quickly returning to the area after the hemorrhage and is often referred to as cardiovascular and brain negligence. Accordingly, a subject of the present invention is the use of HDL for the manufacture of a medicament for the prevention and/or treatment of blood loss or reperfusion injury, in particular, HDL can be used for the prevention and/or treatment of a stroke selected from the group consisting of an episode of an episode of an episode For example, organ damage, cardiac arrhythmia, cardiac reperfusion injury, and organ transplants such as kidney, heart and liver or heart-lung catheter surgery and other conditions, even more surprised, found that HDL is There is a beneficial effect when a temporary or permanent closure occurs, so that the closure or removal of a closed blood clot or other entity is not a prerequisite for efficacy, and the HDL is still displayed 6 or more hours after the event of an anemia Useful, another surprising observation is that the use of HDL before the event of an anemia has a beneficial effect. Other embodiments of the present invention are directed to the use of HDL for the prevention and/or treatment of temporary episodes of episodes (TIA), which are common and about one-third will develop a stroke in the future. The most common cause of TIA is from large Atherosclerotic plaques in blood vessels (usually narrow atherosclerotic carotid arteries) form thrombi because HDL has anti-atherosclerotic properties, as explored by endothelial resonance through resonance of bioavailability of nitric oxide It is shown that in the adjustment of vascular tone and structure (9), Xianxin HDL can stabilize the atherosclerotic patch of TIAs, thus reducing the risk of major strokes. Currently used in this paper scale, the Chinese national standard (CNS) ) Α 4 specifications (210X297 mm) (Please read the notes on the back and fill out this page) I·Installation.
、1T 經濟部智慧財產局員工消費合作社印製 -7- 1311485 A7 _________ 五、發明説明（5) (請先閲讀背面之注意事項再填寫本頁} TIAs之醫療包括抗血小板療法、阿斯匹靈、氯苄噻哌啶及 手術介入例如動脈內膜術，但是，這些沒有一個可以實質 上提供降低發病率。 另一個具體實施例是關於在危險的病人族群例如進行 手術的病人中預防性用藥HDL ,用藥HDL可以減低新中風 之發作及/或嚴重度，預防性用藥HDL也可用在有TIAs、動 脈纖維化及無症狀的頸動脈狹窄之病人。 HDL用於治療上述疾病之用途，特別是用於治療中風 及臨時絕血發作之用途，實踐目前未達成的臨床需求，其 提供臨床有效的醫療用於創傷性腦部傷害之個人。 名詞”HDL”在本發明使用時係關於類似高密度脂蛋白之 粒子且含初生的HDL或再構建HDL(rHDL)或其任何混合 物，此粒子可從蛋白質或肽成份及從脂質製造，名詞”HDL” 也包括任何再構建高密度脂蛋白。 經濟部智慧財產局K工消費合作社印製 蛋白質較宜是載脂蛋白例如人類載脂蛋白或再構建載 脂蛋白，或具有類性性質之肽，合適的脂質是磷脂質，較 宜是磷脂醯膽鹼，視需要混合其他脂質（膽固醇、膽固醇 酯、三甘油酯或其他脂質），脂質可以是合成的脂質、天然 產生的脂質或其再構建物。 用藥HDL可導致一方面在數個臨床參數上觀察到短時 間效應也就是立即有利的效應，且此不僅可發生在中風發 作後3小時內，且即使6小時或可能更久，另一方面在脂 質上可得到長期有利的改變，而且，HDL非常類似體內自 然產生的物質，且因此用藥HDL沒有副作用。 本紙張尺度適用中國國家標準（CNS ) A4规格（2!〇><297公釐） 1311485 A7 B7 五、發明説明（6 ) (請先閱讀背面之注意事項再填寫本頁) HDL較宜經由輸注用藥，例如經由動脈、腹膜內或較 宜靜脈注射及/或足以得到所要的藥理效應之劑量下輸注， 例如HDL可以在絕血開始前用藥（如果可以預期，例如器官 移植前）及/或絕血期間、再灌流前及/或短時間後用藥，較 宜是24小時-48小時內。 HDL劑量範圍較宜從10-200毫克，更宜是40-80毫克 HDL(以載脂蛋白重量爲基準）每公斤體重每次治療，例如 HDL用藥之劑量可以是約20- 1 00毫克HDL每公斤體重（以 載脂蛋白重量爲基準），作爲大九劑藥物注射及/或作爲輸注 供臨床需要的時間提供，例如期間從數分鐘至數小時，例 如達24小時，如果需要時，HDL用藥可以重複一或數次。 再構建高密度脂蛋白（rHDL)可以從人類載脂蛋白A-l(apoA-1)例如從人類血漿分離及大豆衍生的磷脂醯膽鹼（pc) 製備，混合的克分子比例是約1:150 apoA-l:PC。 經濟部智慧財產局員工消費合作社印製 根據本發明，HDL例如初生的HDL、rHDL或類似HDL 的粒子特別較宜，其蛋白質例如載脂蛋白A-1及磷脂質脂 克分子比例範圍是1:50至1:250，特別是約1:150，另外 rHDL可以視需要含其他脂質例如膽固醇、膽固醇酯、三甘 油酯及/或鞘脂類，較宜克分子比例是達1: 20 ,例如1: 5至 1:20，以載脂蛋白爲基準，較佳的rHDL揭示在EP-A-0663 407 ° HDL之用藥可以結合其他藥劑用藥，例如溶血栓劑、 抗發炎劑、神經-及/或心臟保護劑。 而且，本發明也關於用於預防及/或治療絕血或再灌流 本紙張又度適用中國國家標準（CNS ) A4規格（210X297公釐） 1311485 A7 B7 經濟部智慧財產局員工消費合作社印製 五、發明説明（7) 傷害之醫藥組成物，其中含有效量之HDL,較宜HDL是用 藥至人類病人。 另外，本發明將經由下列實例詳細說明： 實例1 興奮中毒損傷： 在用水合氯醛（400毫克/公斤，腹膜內）麻醉的Sprague-Dawley大鼠進行實驗，將股靜脈連接導管供輸注HDL，將 大鼠放在立體定位的裝置內且在，且經中線切口後，接受 單側注射N-甲基-D-天冬胺酸（NMDA)或媒劑進入右溝，座 標：〇.2毫米後面，3毫米側面，5.5毫米腹面至前囪，插入 針後5分鐘，使用Hamilton針栗在0,5毫升/分鐘之速率下 注射溶液歷經5分鐘，完成注射後5分鐘，取出注射針。 在此系列實驗中，大鼠接受靜脈輸注食鹽水（n=5)(5微 升/分鐘）經4小時，經2小時後，進行單側注射NMD A (75 毫微莫耳濃度在3毫升磷酸鹽緩衝化的食鹽水中，pH 7.4) 至右溝，經24小時後，將大鼠殺死並取出腦供組織學分 析，在另一組實驗中，大鼠在1 20毫克/公斤之劑量下接受 靜脈輸注rHDL (n = 5)(5微升/分鐘）經4小時，經2小時後， 進行單側注射NMDA(75毫微莫耳濃度在3毫升磷酸鹽緩衝 化的食鹽水中，pH 7.4)至右溝並持續靜脈輸注rHDL經2小 時，經24小時後，將大鼠殺死並取出腦供組織學分析，結 果列在表1。 (請先閲讀背面之注意事項再填寫本頁)1T Ministry of Economic Affairs Intellectual Property Bureau Staff Consumer Cooperative Printed -7- 1311485 A7 _________ V. Invention Description (5) (Please read the notes on the back and fill out this page) TIAs medical treatment includes antiplatelet therapy, aspirin , benzyl thiophene piperidine and surgical intervention such as endarterial surgery, however, none of these can substantially provide a reduction in morbidity. Another specific embodiment relates to prophylactic HDL in a patient population at risk, such as surgery. HDL can reduce the onset and/or severity of new strokes. Preventive medication HDL can also be used in patients with TIAs, arterial fibrosis, and asymptomatic carotid stenosis. HDL is used for the treatment of these diseases, especially For the treatment of stroke and temporary episodes of seizures, practice the currently unmet clinical need to provide clinically effective medical treatment for individuals with traumatic brain injury. The term "HDL" is used in the context of the present invention for similar high density lipids. Protein particles containing nascent HDL or reconstituted HDL (rHDL) or any mixture thereof, which may be derived from protein or peptide components and from lipids Manufacture, the term "HDL" also includes any re-construction of high-density lipoprotein. The Ministry of Economic Affairs, the Intellectual Property Bureau, K-Consumer Cooperative, prints proteins that are preferably apolipoproteins such as human apolipoproteins or reconstituted apolipoproteins, or have classes. A peptide of a sexual nature, a suitable lipid is a phospholipid, preferably a phospholipid choline, optionally mixed with other lipids (cholesterol, cholesterol ester, triglyceride or other lipid), the lipid may be a synthetic lipid, a naturally occurring lipid Or its reconstitution. The use of HDL can result in the observation of short-term effects, ie immediate beneficial effects, on several clinical parameters, and this can occur not only within 3 hours after the onset of stroke, but even 6 hours or possibly more For a long time, on the other hand, long-term favorable changes can be obtained on the lipid, and HDL is very similar to the naturally occurring substances in the body, and therefore the HDL has no side effects. The paper scale applies to the Chinese National Standard (CNS) A4 specification (2!〇> ; <297 mm) 1311485 A7 B7 V. Invention Description (6) (Please read the note on the back and fill in this page) HDL is better Injectable, for example, via arterial, intraperitoneal or intravenous injection and/or infusion at a dose sufficient to achieve the desired pharmacological effect. For example, HDL can be administered prior to the onset of hemostasis (if expected, for example, prior to organ transplantation) and/or During the period of anemia, before reperfusion and/or after a short time, it is more suitable to be within 24 hours to 48 hours. The HDL dose range is preferably from 10 to 200 mg, more preferably 40 to 80 mg of HDL (with apolipoprotein weight) For the basis of each treatment per kg body weight, for example, the dose of HDL can be about 20-100 mg HDL per kg body weight (based on the weight of apolipoprotein), as a large nine-drug injection and / or as an infusion for clinical The time required is provided, for example, from a few minutes to several hours, for example up to 24 hours, and if desired, the HDL medication can be repeated one or several times. Reconstruction of high-density lipoprotein (rHDL) can be prepared from human apolipoprotein A1 (apoA-1), for example, from human plasma and soybean-derived phospholipid choline (pc). The molar ratio of the mixture is about 1:150 apoA. -l:PC. According to the present invention, HDL such as nascent HDL, rHDL or HDL-like particles are particularly preferred, and the ratio of proteins such as apolipoprotein A-1 and phospholipid lipids is 1: 50 to 1:250, especially about 1:150. In addition, rHDL may contain other lipids such as cholesterol, cholesterol ester, triglyceride and/or sphingolipid as needed, and the ratio of the molar ratio is 1:20, for example, 1 : 5 to 1:20, based on apolipoprotein, the preferred rHDL revealed that the drug in EP-A-0663 407 ° HDL can be combined with other drugs such as thrombolytics, anti-inflammatory agents, nerves and/or hearts. Protective agent. Moreover, the present invention also relates to the use of the Chinese National Standard (CNS) A4 specification (210X297 mm) for the prevention and/or treatment of blood-stable or reperfusion paper. 1311485 A7 B7 Ministry of Economic Affairs Intellectual Property Bureau employee consumption cooperative printed five (Invention) (7) The medical composition of the injury, which contains an effective amount of HDL, is preferably administered to a human patient. In addition, the present invention will be described in detail by the following examples: Example 1 Excitotoxic damage: Experiments were performed on Sprague-Dawley rats anesthetized with chloral hydrate (400 mg/kg, ip), and the femoral vein connecting catheter was used for infusion of HDL. The rats were placed in a stereotactic device and, after a midline incision, received a unilateral injection of N-methyl-D-aspartate (NMDA) or vehicle into the right groove, coordinates: 〇.2 Behind the millimeter, 3 mm side, 5.5 mm ventral to the bregma, 5 minutes after insertion of the needle, the solution was injected using Hamilton needles at a rate of 0,5 ml/min for 5 minutes, and 5 minutes after the injection was completed, the needle was removed. In this series of experiments, rats received intravenous infusion of saline (n=5) (5 μl/min) over 4 hours, and after 2 hours, unilateral injection of NMD A (75 mM molar concentration in 3 ml) In phosphate-buffered saline, pH 7.4) to the right ditch, after 24 hours, the rats were sacrificed and the brain was removed for histological analysis. In another set of experiments, the rats were dosed at 20 mg/kg. Underwent intravenous infusion of rHDL (n = 5) (5 μl/min) over 4 hours, and after 2 hours, unilateral injection of NMDA (75 mM molar concentration in 3 ml phosphate buffered saline, pH) 7.4) To the right sulcus and continuous intravenous infusion of rHDL for 2 hours. After 24 hours, the rats were sacrificed and the brain was removed for histological analysis. The results are shown in Table 1. (Please read the notes on the back and fill out this page)
本紙張尺度適用中國國家標準（CNS ) A4規格（210X297公釐） -10 - 1311485 A7 B7 五、發明説明（8 ) 表1 :損傷體積（立方毫米） 大鼠 對照組 rHDL 1 50.27 16.54 2 47.05 18.86 3 41.28 17.44 4 38.5 17.51 5 51.66 19.86 N 5 5 平均 45.75 18.04 SD 5.69 1.31 SEM 2.55 0.59 (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 在此實驗中，與對照組比較，在rHDL處理的動物中腦 壞死體積大幅降低60.6%。 在另一個系列之實驗中，在NMDA注射經3小時後， 輸注用藥rHDL( 120毫克/公斤）或安慰劑（食鹽水）經4小 時，經24小時後組織學測量血梗大小，結果列在表2 ° 本紙張尺度適用中國國家椟準（CNS > A4規格（210X297公釐） -11 - 1311485 經濟部智慧財產局g(工消費合作社印製 五、發明説明（9) 表2 _ . 食鹽水+NMDA rHDL+NMDA _ 損傷體積（立方毫米） 損傷體積（立方毫米） 175 77 101 83 105 133 180 121 149 51 115 66 平均 137 88 SD 35 32 降低％ -36% P(學生t測試） 0.03 在此實驗中，發現血梗大小降低36%。 (請先閱讀背面之注意事項再填寫本頁) 實例2 中間腦動脈閉合： 2.1閉合前用藥 在用水合氯醛（400毫克/公斤，腹膜內）麻醉的Sprague-Dawley大鼠進行實驗，將氣管連接導管並將動物機械式通 風空氣並補充氧氣使血液氣體維持在正常範圍內，連續監 測直腸溫度並維持在37°C,將將導管放入股動脈以測量全 身血壓及監視血液氣體，將股靜脈連接導管供輸注藥劑， 進行頸中線切口並暴露右頸總動脈，其分枝凝固後，將外 本紙張尺度適用中國國家檁準（CNS ) A4規格（210X297公釐） -12- 1311485 A7 B7 經濟部智慧財產局員工消費合作社印製 五、發明説明（1〇) 頸動脈（EAC)末梢部打開，在ECA之腔內插入含熱熔膠矽（2 毫米長，直徑0.3 8毫米）末梢圓柱體之尼龍線（直徑0.22毫 米）並前進至內頸動脈之MCA源頭，爲了恢復MCA血液流 動，在30分鐘後將尼龍線取出並剪斷。 組織學分析：經24小時後進行手術安樂死，快速將腦 取出，在-50°C之異戊烷中冷凍並儲存在-8(TC，用硫堇染色 冰凍切片的頭冠腦部份（20微米）並用影像分析儀分析，與 健康組織之顏色比較，經由變化組織中組織學染色之倉白 色界定損傷區域，經由使用定向寰椎測定大鼠之有價値區 域並使用影像分析系統測量損傷區域。 在此系列實驗中，大鼠接受靜脈輸注鹽水（n = 5)(5微升/ 分鐘）經4小時，經2小時後將大鼠之MCA閉合30分鐘後 再灌流，經24小時後，將大鼠殺死用於腦之組織學分析， 在另一組實驗中，大鼠在1 20毫克/公斤之劑量下接受靜脈 輸注rHDL (n=5)(5微升/分鐘）經4小時，經2小時後將大鼠 之MCA閉合30分鐘後再灌流，經24小時後，將大鼠殺死 用於腦之組織學分析，結果列在表3。 在MCA閉合模式中，得到下列結果： (請先閱讀背面之注意事項再填寫本頁) 本紙張尺度適用中國國家標準（CNS ) A4規格（210X297公釐） -13- 1311485 A7This paper scale applies to Chinese National Standard (CNS) A4 specification (210X297 mm) -10 - 1311485 A7 B7 V. Description of invention (8) Table 1: Damage volume (cubic mm) Rat control group rHDL 1 50.27 16.54 2 47.05 18.86 3 41.28 17.44 4 38.5 17.51 5 51.66 19.86 N 5 5 Average 45.75 18.04 SD 5.69 1.31 SEM 2.55 0.59 (Please read the note on the back and fill out this page) The Ministry of Economic Affairs Intellectual Property Bureau employee consumption cooperative is printed in this experiment, Compared with the control group, the volume of brain necrosis was significantly reduced by 60.6% in the animals treated with rHDL. In another series of experiments, 3 hours after NMDA injection, the infusion of rHDL (120 mg / kg) or placebo (salt) was administered for 4 hours. After 24 hours, histology was measured by hematology. The results were listed in Table 2 ° The paper scale is applicable to China National Standard (CNS > A4 specification (210X297 mm) -11 - 1311485 Ministry of Economic Affairs Intellectual Property Bureau g (Working Consumer Cooperative Printing 5, Invention Notes (9) Table 2 _ . Salt Water + NMDA rHDL + NMDA _ Damage volume (cubic mm) Damage volume (cubic mm) 175 77 101 83 105 133 180 121 149 51 115 66 Average 137 88 SD 35 32 % reduction - 36% P (student t test) 0.03 In this experiment, the size of the stalk was found to be reduced by 36%. (Please read the notes on the back and fill out this page.) Example 2 Intermediate cerebral artery closure: 2.1 Pre-closure medication in chloral hydrate (400 mg/kg, intraperitoneal) Anesthetized Sprague-Dawley rats were tested, the trachea was connected to the catheter and the animals were mechanically ventilated with air and oxygen was added to maintain the blood gas within the normal range. The rectal temperature was continuously monitored and maintained at 37 ° C. The catheter is placed in the femoral artery to measure the whole body blood pressure and monitor the blood gas. The femoral vein is connected to the catheter for infusion, the neckline incision is made and the right common carotid artery is exposed. After the branch is solidified, the external paper scale is applied to the Chinese national standard. Quasi (CNS) A4 specification (210X297 mm) -12- 1311485 A7 B7 Ministry of Economic Affairs Intellectual Property Bureau employee consumption cooperative printing 5, invention description (1〇) Carotid artery (EAC) distal opening, inserted in the cavity of ECA A nylon thread (0.22 mm diameter) containing a hot-melt capsule (2 mm long, 0.38 mm diameter) distal cylinder and advanced to the MCA source of the internal carotid artery. To restore MCA blood flow, remove the nylon thread after 30 minutes. Histological analysis: After 24 hours, the operation was euthanized, the brain was quickly taken out, frozen in isopentane at -50 ° C and stored at -8 (TC, stained with frozen sulphur. Part (20 micron) and analyzed by image analyzer, compared with the color of healthy tissue, the lesion area was defined by the white color of histological staining in the changed tissue, and the rat was determined by using the directional vertebrae. The sputum area was measured using an image analysis system. In this series of experiments, rats received intravenous infusion of saline (n = 5) (5 μl/min) for 4 hours, and after 2 hours, the MCA of the rats was closed 30 After a minute, the rats were perfused. After 24 hours, the rats were sacrificed for histological analysis of the brain. In another group, the rats received intravenous infusion of rHDL at a dose of 20 mg/kg (n=5). (5 μl/min) After 4 hours, the MCA of the rats was closed for 30 minutes and then perfused. After 24 hours, the rats were sacrificed for histological analysis of the brain. The results are shown in Table 3. . In the MCA closed mode, the following results are obtained: (Please read the notes on the back and fill out this page.) This paper size applies to the Chinese National Standard (CNS) A4 specification (210X297 mm) -13- 1311485 A7
五、發明説明（W :損傷體積（立方毫米） ___ 大鼠 對照組 rHDL _____ 1 158.94 54.18 —_ 2 229.78 35.27 — 3 201.52 37.64 — 4 193.02 34.64 5 210.24 76.74 _ η 5.00 5.00 _ 平均 198.70 47.69 SD — 26.08 18.11 SEM 11.66 8.10 -----：---：----- (請先閱讀背面之注意事項再填寫本頁) 訂 經濟部智慧財產局員工消費合作社印製 與對照組大鼠比較，rHDL降低腦壞死體積達76%。 2.2閉合後用藥 在MCAo模式中，手術3小時後用藥rHDL，經由插入 尼龍線通過頸動脈並在30分鐘後恢復血液流動，經3小時 後，其接受靜脈輸注rHDL (120毫克/公斤經4小時，6毫 升/公斤經4小時）或鹽水（6毫升/公斤經4小時），將大鼠隨 意指定爲rHDL或未照組，另四隻大鼠進行相同步驟之 MCA閉合，但是尼龍線停在內頸動脈，沒有干擾頸動脈血 液流動並在30分鐘後取出（Sham MCAo組），經3小時後在 靜脈內將2隻此組的大鼠接受rHDL且2隻接受鹽水（6毫升 /公斤經4小時），經24小時後，將全部大鼠殺死並取出腦 本紙張尺度適用中國國家標準（CNS ) A4規格（210X297公釐） -14- 1311485 A7 — B7 五、發明説明（12) 用於組織學分析，與健康組織之顏色比較，經由變化組織 中組織學染色之倉白色界定損傷區域，經由使用定向寰椎 測定大鼠之有價値區域並使用影像分析系統（NIH影像）測量 損傷區域。 在Sham MCAo組沒有損傷。 其他12隻大鼠在MCA閉合後，靜脈內用藥鹽j rHDL，從影像分析之結果呈現在表4，結果顯示閉合@ 小時輸注rHDL導致血梗體積（立方毫米）減少60%。 經濟部智慧財產局員工消費合作社印製 (請先閱讀背面之注意事項再填寫本頁j 表4 損傷體積（立方毫米） 對照組 〜------ 姻L 大鼠 1 88.94 -^^ 87 2 118.9 -------------------_ 46.9] 3 110.06 ----- 43.91 4 121.09 ^^------- 43.13 5 224.14 36.65 6 157.45 35.63 平均 、、、 SD 136.8 48.9 48.2 19.2 減少％ 64% P(學生t測試） 〇-〇〇2〇 本紙張尺度適用中國國家標準（CNS ) A4規格{ 2I0X297公釐） -15- 1311485 A7 B7 五、發明説明（13) 與對照組大鼠比較，壞死體積降低64%。 (請先閱讀背面之注意事項再填寫本頁) 結論：在兩種模式中，與用安慰劑處理的對照組比 較，在rHDL處理的動物中見到血梗體積大幅下降：興奮中 毒模式：60.6%或36%降低壞死體積；MCA閉合模式：降 低 76% 或 60%。 實例3 在大鼠模式中用藥rHDL用於中風（MCA閉合模式） 方法 在此硏究中使用120隻雄性Sprague-Dawley大鼠，1〇〇 隻大鼠接受暫時閉合或永久閉合，20隻大鼠作爲手術及 rHDL對照組，引發中風前2小時或3或6小時後輸注rHDL (120毫克/公斤/4小時），相同於實例4使用之線閉合方法。 將大鼠分成三組，第1組在接受暫時MCA閉合（2小 時）2小時前接受預防劑量之rHDL並在閉合期間連續接受處 理，然後將動脈再灌注。 經濟部智慧財產局員工消費合作社印製 第2組在再灌注後接受暫時MCA閉合，在3小時或6 小時後提供處理。 第3組接受永久MCA閉合並在閉合3小時或6小時後 接受處理。 經過上述議案後，使用稱爲前肢彎曲、軀幹丨丑_ 個| 推及移動性之四種標準的用動神經測試法檢查神經變{匕； 分數加入各測試且結果呈現在圖1。 從此圖明顯地指出rHDL提供作爲前處理及作爲閉合 本紙張尺度適用中國國家標準（CNS ) A4規格（210X297公釐> -16- 1311485 A7 !_-______B7_ 五、發明説明（14) (暫時及永久）後3或6小時給藥，導致優於未經處理的大鼠 之較佳神經系統分數。 神經系統分析後，將大鼠殺死並將腦取出，使用經陽 光反射測量血梗面積之衝擊光技術檢視大鼠腦部，用HDL 處理的永久及暫時MCAo之結果顯示在圖2及3。 這些圖顯示如果rHDL提供至大鼠⑴閉合前2小時，總 血梗體積降低54% (ii)暫時閉合後3小時，降低65%且（iii) 暫時閉合後6小時，降低62% ,永久閉合在兩個處理時間 都觀察到59%之類似下降。 據此，用藥rHDL在閉合前可有效地作爲預防處理，且 在閉合後兩個不同的時間點可作爲醫療處理，更確定地 說，預防及醫療處理可以結合。 (請先閲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 本紙張尺度適用中.國國家梂準（CNS ) A4規格（210X297公釐） -17- 經濟部智慧財產局員工消費合作杜印製 1311485 A7 ______B7五、發明説明（15) 文獻 1. American Heart Association (AHA), 2000 2. Hays SJ. Therapeutic approaches to the treatment of neuroinflammatory diseases. Curr Pharm Des 4:335-348, 1998 3. Jean WC, Spellman SR, Nussbaum ES, Low WC, Reperfusion injury after focal cerebral ischemia: the role of inflammation and the therapeutic horizon. Neurosurgery 43:1382-1396, 1998 4. Barone FC, Feuerstein GZ. Inflammatory mediators and stroke: new opportunities for novel therapeutics. J Cereb Blood Flow Metab 19:819-834, 1999 5. Morgan WM, and O’Neill JA. Hemorrhagic and obstructive shock in pediatric patients, New Horiz 6: 150-154, 1998 6. Demetriades D, Smith JS, Jacobson LE, Moncure M, Minei J, Nelson BJ and Scannon PJ. Bactericidal/permeability-increased protein (rBPT21) in patients with hemorrhage due to trauma: results of a multicancer phase II clinical trial. RBP121 Acute Hemorrhage Trauma Study Group, J Trauma 46: 667-676, 1999 7. Regel G, Gotz M, Weltner T, Sturm JA and Tscherne H. Pattern of organ failure following severe trauma. World J Surg 20: 422-429, 1996 8. Cryer HG. Therapeutic approaches for clinical ischaemia and reperfusion injury. Shock 8: 26-32, 1997 9. Spieker LE, Sudano 1, Lerch PG, Lang MG, Binggeli C, Corti R, Luscher TF, Noll G. High-density lipoprotein restores endothelial function in hypercholesterolemic men. N Engl J Med, 2000. (in preparation) 10. Feuerstein GZ, Wang X, Barone FC. 丁he role of cytokines in the neuropathologyof stroke and neurotrauma. Neuroimmunomodulation 5:143-159, 1998 11. DeGraba TJ. The role of inflammation after acute stroke: utility of pursuing anti-adhesion molecule therapy. Neurology 51:62-68, 1998 (讀先閱讀背面之注意事項再填寫本頁) -裝· 訂 • n HI i^i 本紙張尺度適用中國國家標準（CNS ) A4規格（210X297公釐） -18- 1311485 A7 B7 五、發明説明（16) 12. Benveniste ΕΝ. Cytokine actions in the central nervous system. Cytokine Growth Factor Rev 9:259-275, 1998 13. Van Wagoner NJ, Benveniste EN_ lnterleukin-6 expression and regulation in astrocytes. J. Neuroimmunol 100:124-139, 1999 14. Touzani 0, Boutin H, Chuquet J, Rothwell N. Potential mechanism of interleukin-1 involvement in cerebral ischemia. J Neuroimmunol 100:203-215, 1999 15. del Zoppo G, Ginis I, Hallenbeck JMy ladecola C, Wang X, Feuerstein GZ. Inflammation and stroke: putative role for cytokines, adhesion molecules and iNOS in brain responses to ischemia. Brain Pathol 10:95-112, 2000 16· Du C, R Hu, CA Csernansky, CY Hsu, DW Choi. Very delayed infarction after mild focal cerebral ischemia: A role for apoptosis? J Cereb Blood Flow Metab 16:195-201, 1996 17. Matsuda Y, Hirata K, Inoue N, Suematsu M, Kawahima S, Akita H, Yokoyama M. High density lipoprotein reverses inhibitory effect of oxidized low density lipoprotein on endothelium-dependent arterial relaxation. Circ Res 72(5):1103-1109, 1993 18. Chander R, Kapoor NK. High density lipoprotein is a scavanger of superoxide anions. Biochem Pharmacol 40(7):1663-1665, 1990 19. Araujo FB, Barbosa DS, Hsin CY, Maranhao RC, ^bdalla DS. Evaluation of oxidative stress in patients with hyperlipidemia. Atherosclerosis 117(1):61-71, 1995 20. Huang JMf Huang ZX, Zhu W. Mechanism of high-density lipoprote in subtractions inhibiting copper-catalyzed oxidation of low-density lip叩「otein. Clin Biochem 31(7):537-543, 1998 21. Cockerill GW et al. High-density lipoproteins rescue end-stage organ failure in a rat model of haemorraghic shock. J. Submicroscopic Cyt. Path. 32(3): 353, 2000 本紙張尺度適用中國國家標準（CNS ) A4規格（210X297公釐） (請先閲讀背面之注意事項再填寫本頁) •裝· 訂 經濟部智慧財產局員工消費合作社印製 -19 -V. INSTRUCTIONS (W: Damage volume (cubic millimeters) ___ Rat control group rHDL _____ 1 158.94 54.18 —_ 2 229.78 35.27 — 3 201.52 37.64 — 4 193.02 34.64 5 210.24 76.74 _ η 5.00 5.00 _ Average 198.70 47.69 SD — 26.08 18.11 SEM 11.66 8.10 -----:---:----- (Please read the notes on the back and fill out this page) The Ministry of Economic Affairs, Intellectual Property Bureau, Staff Consumer Cooperatives, printed and compared with the control group, rHDL reduced brain necrosis volume by 76%. 2.2 After closure, the drug was administered in MCAo mode, and rHDL was administered 3 hours after surgery. The carotid artery was inserted through the nylon thread and blood flow was restored after 30 minutes. After 3 hours, it was intravenously infused. Rats were randomly assigned to rHDL or untreated groups with rHDL (120 mg/kg for 4 hours, 6 ml/kg for 4 hours) or saline (6 ml/kg for 4 hours), and the other four rats performed the same procedure. The MCA was closed, but the nylon thread was stopped in the internal carotid artery, did not interfere with carotid blood flow and was taken out after 30 minutes (Sham MCAo group). After 3 hours, 2 rats of this group received intravenous rHDL and 2 Only saline (6 ml / kg for 4 hours) is received. After 24 hours, all rats are killed and the brain is taken out. The Chinese National Standard (CNS) A4 specification (210X297 mm) -14-1311485 A7 - B7 V. INSTRUCTIONS (12) For histological analysis, compared with the color of healthy tissue, the lesion area is defined by the histological staining white in the changing tissue, and the valuable sputum area of the rat is determined by using the directional vertebrae and the image is used. The analysis system (NIH image) measured the lesion area. There was no damage in the Sham MCAo group. The other 12 rats were intravenously administered with the drug j rHDL after the MCA was closed. The results from the image analysis are shown in Table 4. The results showed a closed @ hour infusion. rHDL causes a 60% reduction in blood vessel volume (cubic millimeters). Printed by the Ministry of Economic Affairs Intellectual Property Bureau employee consumption cooperative (please read the notes on the back and fill out this page again. j Table 4 Damage volume (cubic millimeters) Control group~--- --- Marriage L Rat 1 88.94 -^^ 87 2 118.9 -------------------_ 46.9] 3 110.06 ----- 43.91 4 121.09 ^^- ------ 43.13 5 224.14 36.65 6 15 7.45 35.63 Average,,, SD 136.8 48.9 48.2 19.2 Reduction % 64% P (student t test) 〇-〇〇2〇 This paper scale applies to Chinese National Standard (CNS) A4 specification { 2I0X297 mm) -15- 1311485 A7 B7 V. INSTRUCTIONS (13) Compared with the control group, the necrotic volume was reduced by 64%. (Please read the precautions on the back and fill out this page.) Conclusion: In both modes, a significant decrease in blood vessel volume was observed in rHDL-treated animals compared to placebo-treated controls: Excitotoxicity pattern: 60.6 % or 36% reduced necrotic volume; MCA closed mode: reduced by 76% or 60%. Example 3 Administration of rHDL in rat mode for stroke (MCA closed mode) Method 120 male Sprague-Dawley rats were used in this study, 1 rat received temporary closure or permanent closure, 20 rats As a surgical and rHDL control group, rHDL (120 mg/kg/4 hours) was infused 2 hours or 3 or 6 hours prior to stroke, as in the line closure method used in Example 4. The rats were divided into three groups, and the first group received a prophylactic dose of rHDL 2 hours before receiving the temporary MCA closure (2 hours) and was continuously treated during the closure period, and then the artery was reperfused. Ministry of Economic Affairs, Intellectual Property Office, Staff Consumer Cooperative Printed Group 2 received temporary MCA closure after reperfusion and provided treatment after 3 or 6 hours. Group 3 received a permanent MCA closure and was treated 3 hours or 6 hours after closure. After the above motion, the nerves were examined using a four-standard method called forelimb bending, torso 丨 _ _ | push and mobility. The scores were added to each test and the results are presented in Figure 1. From this figure it is clearly pointed out that rHDL is provided as a pre-treatment and as a closed paper scale applicable to the Chinese National Standard (CNS) A4 specification (210X297 mm> -16-1311485 A7 !_-______B7_ V. Invention Description (14) (temporary and Administration 3 or 6 hours after permanent) resulted in better neurological scores than in untreated rats. After neurological analysis, the rats were sacrificed and the brain was removed, and the area of the blood incision was measured using sunlight reflection. The results of percutaneous light techniques were used to examine the brain and the results of permanent and transient MCAo treatment with HDL are shown in Figures 2 and 3. These figures show that if rHDL is supplied to rats (1) 2 hours before closure, the total stenosis volume is reduced by 54% (ii) ) 3 hours after temporary closure, 65% reduction and (iii) 6 hours after temporary closure, 62% reduction, permanent closure observed a similar decrease of 59% at both treatment times. According to this, the drug rHDL can be effective before closure. As a preventive treatment, it can be used as a medical treatment at two different time points after closure. More specifically, prevention and medical treatment can be combined. (Please read the notes on the back and fill out this page.) Ministry of Economic Affairs Bureau employees' consumption cooperatives print this paper scale applicable to the country's national standard (CNS) A4 specifications (210X297 mm) -17- Ministry of Economic Affairs Intellectual Property Bureau employees consumption cooperation Du printing 1311485 A7 ______B7 V. Invention description (15) 1. American Heart Association (AHA), 2000 2. Hays SJ. Therapeutic approaches to the treatment of neuroinflammatory diseases. Curr Pharm Des 4:335-348, 1998 3. Jean WC, Spellman SR, Nussbaum ES, Low WC, Reperfusion Injury after focal cerebral ischemia: the role of inflammation and the therapeutic horizon. Neurosurgery 43:1382-1396, 1998 4. Barone FC, Feuerstein GZ. Inflammatory mediators and stroke: new opportunities for novel therapeutics. J Cereb Blood Flow Metab 19:819 -834, 1999 5. Morgan WM, and O'Neill JA. Hemorrhagic and obstructive shock in pediatric patients, New Horiz 6: 150-154, 1998 6. Demetriades D, Smith JS, Jacobson LE, Moncure M, Minei J, Nelson BJ and Scannon PJ. Bactericidal/permeability-increased protein (rBPT21) in patients with hemorrhage due to trauma RBP121 Acute Hemorrhage Trauma Study Group, J Trauma 46: 667-676, 1999 7. Regel G, Gotz M, Weltner T, Sturm JA and Tscherne H. Pattern of organ failure following severe trauma World J Surg 20: 422-429, 1996 8. Cryer HG. Therapeutic approaches for clinical ischaemia and reperfusion injury. Shock 8: 26-32, 1997 9. Spieker LE, Sudano 1, Lerch PG, Lang MG, Binggeli C, Corti R, Luscher TF, Noll G. High-density lipoprotein restores endothelial function in hypercholesterolemic men. N Engl J Med, 2000. (in preparation) 10. Feuerstein GZ, Wang X, Barone FC. Ding he role of cytokines in the neuropathologyof Stroke and neurotrauma. Neuroimmunomodulation 5: 143-159, 1998 11. DeGraba TJ. The role of inflammation after acute stroke: utility of pursuing anti-adhesion molecule therapy. Neurology 51:62-68, 1998 (Read the first note on the back) Fill in this page again - Install · Book • n HI i^i This paper scale applies to China National Standard (CNS) A4 specification (210X297 PCT) -18- 1311485 A7 B7 V. INSTRUCTIONS (16) 12. Benveniste ΕΝ. Cytokine actions in the central nervous system. Cytokine Growth Factor Rev 9:259-275, 1998 13. Van Wagoner NJ, Benveniste EN_ lnterleukin-6 Expression and regulation in astrocytes. J. Neuroimmunol 100: 124-139, 1999 14. Touzani 0, Boutin H, Chuquet J, Rothwell N. Potential mechanism of interleukin-1 involvement in cerebral ischemia. J Neuroimmunol 100:203-215, 1999 15. del Zoppo G, Ginis I, Hallenbeck JMy ladecola C, Wang X, Feuerstein GZ. Inflammation and stroke: putative role for cytokines, adhesion molecules and iNOS in brain responses to ischemia. Brain Pathol 10:95-112, 2000 16· Du C, R Hu, CA Csernansky, CY Hsu, DW Choi. Very delayed infarction after mild focal cerebral ischemia: A role for apoptosis? J Cereb Blood Flow Metab 16:195-201, 1996 17. Matsuda Y, Hirata K, Inoue N, Suematsu M, Kawahima S, Akita H, Yokoyama M. High density lipoprotein reverses inhibitory effect of oxidized low density lipoprotein on Endothelium-dependent arterial relaxation. Circ Res 72(5): 1103-1109, 1993 18. Chander R, Kapoor NK. High density lipoprotein is a scavanger of superoxide anions. Biochem Pharmacol 40(7): 1663-1665, 1990 19. Aaurosclerosis 117(1):61-71, 1995 20. Huang JMf Huang ZX, Zhu W. Mechanism of high- Density lipoprote in subtractions inhibiting copper-catalyzed oxidation of low-density lip叩"otein. Clin Biochem 31(7):537-543, 1998 21. Cockerill GW et al. High-density lipoproteins rescue end-stage organ failure in a rat Model of haemorraghic shock. J. Submicroscopic Cyt. Path. 32(3): 353, 2000 This paper scale applies to Chinese National Standard (CNS) A4 specification (210X297 mm) (please read the notes on the back and fill out this page) • Installation · Ministry of Economic Affairs Intellectual Property Bureau employee consumption cooperative printing -19 -