TWI267376B - Isatin compounds and pharmaceutical composition comprising the same - Google Patents

Abstract

The invention provides an isatin compound having formula (I), wherein R1 is selected from the group consisting of hydrogen, halogen, cyano, amide, and nitro; X is a C1-C5 alkyl, alkenyl, or alkynyl; and Het is a substituted or non-substituted heterocycle group. The invention also provides a pharmaceutical composition including the compound.

Description

1267376 九、發明說明： 【發明所屬之技術領域】 本發明係有關於一種化合物，特別是有關於一種用於抑制 SARS冠狀病毒主要蛋白水解酵素活性之吲哚醌化合物。 【先前技術】 2003年初爆發的肺炎，已由國際衛生組織（World Health Organization, WHO)定義為”急性呼吸道症候群（severe acute respiratory syndrome，SARS)”，又稱非典型肺炎。當時在香港及 世界多個地區包括中國、越南、新加坡、加拿大、台灣及美國 等地相繼發現病例，前五個地方更有死亡個案出現，死亡率達 4%。SARS是一種由病毒所導致的呼吸疾病，根據世界衛生組 織（WHO)指出，初步研究顯示導致此疾病的主要病源是新變種 的冠狀病毒（Coronavirus)，但病毒致病原因仍有待研究，其潛伏 期約為二至七天，而美國疾病及預防中心（CDC)更指出有潛伏期 長達十日左右的個別病例。SARS的主要症狀包括：發熱（38°C)、 乾咳、呼吸急速、呼吸困難、肺部X光檢驗顯示肺炎徵狀，或 如頭痛、肌肉寒顫、食欲不振、精神錯亂、出紅疹、肚瀉等病 狀。 人類冠狀病毒（human cor〇navirus，HCoV)已被證實為引發 SARS的感染源，冠狀病毒為正股RNA(positive-stranded RNA) 病毒，其特色是至目前為止最大的病毒RNA genomes。SARS 病毒圓環上和外殼表面分佈著大大小小的蛋白質，其中對感染 患病有重要影響的可能是以下六種關鍵蛋白，包括E蛋白、S 蛋白、Μ蛋白、N蛋白、多聚酶和3CL蛋白水解酶。當SARS 冠狀病毒在複製（replication)及轉錄（translation)過程中，需要蛋 0296-A20607TWF(N2)；david 6 1267376 白質水解酵素（main proteinase Mpr。，或稱 3C_like proteinase， 3CLpro)將兩個蛋白質（ppla以及pplab)水解成p〇iypeptide。 德國Liibeck大學Hilgenfeld教授的研究小組，在Science 沿cprea 2003，May 13·發表一份研究報告，指出一種人類冠狀 病毒 HCoV 229E 在複製（replication)及轉錄（transcription)過程 中，須要使用一種蛋白水解酵素（main proteinase Μρπ)，或稱 3C_like proteinase 3CLpr。）將蛋白質（ppla and pplab)水解成 polypeptide，且亦發現人類冠狀病毒HCoV 229E、人類普通感 冒鼻病毒（human rhinovirus HRV)以及豬類痛j疾冠狀病毒 (porcine transmissible gastroenteritis coronavirus TGEV)蛋白質 水解酵素（Mpf°)的活性部位（active site)結構極為類似，他們也發 現，人類SARS冠狀病毒蛋白質水解酵素（SARS-HCoV Mpn))的 活性部位，亦有類似結構，由於人類冠狀病毒與感冒病毒具有 類似構造，因此，利用治療感冒病毒現有的藥物加以修改’可 視為開發抑制SARS冠狀病毒藥物的途徑。 【發明内容1 有鑑於此，本發明之目的係揭露一種新穎化合物，用以抑 制SARS冠狀病毒主要蛋白水解酵素之活性。 本發明之另一目的係揭露一種包含該化合物之醫藥组合 物。 為了達成上述目的，本發明係提供一種吲哚醌化合物，具 有化學式（I) 0296-A20607TWF(N2)；david 7 12673761267376 IX. DESCRIPTION OF THE INVENTION: TECHNICAL FIELD OF THE INVENTION The present invention relates to a compound, and more particularly to an indole compound for inhibiting the activity of a major proteolytic enzyme of a SARS coronavirus. [Prior Art] Pneumonia, which broke out in early 2003, has been defined by the World Health Organization (WHO) as "severe acute respiratory syndrome (SARS)", also known as atypical pneumonia. At that time, cases were found in Hong Kong and various parts of the world including China, Vietnam, Singapore, Canada, Taiwan and the United States. In the first five places, there were more deaths and the death rate was 4%. SARS is a respiratory disease caused by viruses. According to the World Health Organization (WHO), preliminary research shows that the main cause of this disease is the new variant Coronavirus, but the cause of the virus remains to be studied, and its incubation period It is about two to seven days, and the Centers for Disease Control and Prevention (CDC) has pointed out that there are individual cases with an incubation period of up to ten days. The main symptoms of SARS include: fever (38 ° C), dry cough, rapid breathing, difficulty breathing, pulmonary X-ray examination showing signs of pneumonia, or such as headache, muscle chills, loss of appetite, confusion, rash, diarrhea And other symptoms. The human cor〇navirus (HCoV) has been shown to be the source of SARS infection, and the coronavirus is a positive-stranded RNA, which is characterized by the largest viral RNA genomes to date. There are large and small proteins distributed on the surface of the SARS virus and on the surface of the outer shell. Among them, the following six key proteins may be important for the infection, including E protein, S protein, prion protein, N protein, polymerase and 3CL protein. Hydrolase. When the SARS coronavirus is involved in replication and translation, it needs egg 0296-A20607TWF (N2); david 6 1267376 white protein hydrolyzing enzyme (main proteinase Mpr., or 3C_like proteinase, 3CLpro) to bind two proteins ( Ppla and pplab) are hydrolyzed to p〇iypeptide. Professor Hilgenfeld's research team at the University of Liibeck in Germany, published a study in Science along cprea 2003, May 13 ·, pointing out that a human coronavirus HCoV 229E requires the use of a proteolytic enzyme during replication and transcription. (main proteinase Μρπ), or 3C_like proteinase 3CLpr. The protein (ppla and pplab) was hydrolyzed into a polypeptide, and human coronavirus HCoV 229E, human rhinovirus HRV, and porcine transmissible gastroenteritis coronavirus TGEV protein hydrolyzing enzyme (also known as porcine transmissible gastroenteritis coronavirus TGEV) were also found. The active site structure of Mpf° is very similar. They also found that the active site of human SARS coronavirus protein hydrolyzing enzyme (SARS-HCoV Mpn) has a similar structure, because human coronavirus has similarity to cold virus. The construction, therefore, is modified by the use of existing drugs for the treatment of cold viruses' as a way to develop drugs that inhibit SARS coronavirus. SUMMARY OF THE INVENTION In view of the above, it is an object of the present invention to disclose a novel compound for inhibiting the activity of a major proteolytic enzyme of a SARS coronavirus. Another object of the invention is to disclose a pharmaceutical composition comprising the compound. In order to achieve the above object, the present invention provides an anthracene compound having the formula (I) 0296-A20607TWF (N2); david 7 1267376

其中I係擇自氫、i素、腈基、醯胺基及硝基所組成之族 群；X係為一碳數1〜5之烷基、烯基或炔基；以及Het係為一 有取代或無取代之雜環基。 本發明另提供一種包含該吲哚醌化合物或其鹽類之醫藥組 合物，該醫藥組合物更包含藥學上可接受之賦形劑（excipient)、 稀釋劑（diluent)或載體（carrier)。 【實施方式】 本發明具有吲哚醌雜環基之化合物，係由下列化學式U) 表示之：Wherein I is selected from the group consisting of hydrogen, i, nitrile, guanamine, and nitro; X is an alkyl, alkenyl or alkynyl group having 1 to 5 carbon atoms; and Het is a substituted group. Or an unsubstituted heterocyclic group. The present invention further provides a pharmaceutical composition comprising the guanidine compound or a salt thereof, the pharmaceutical composition further comprising a pharmaceutically acceptable excipient, diluent or carrier. [Embodiment] The compound of the present invention having an anthracene heterocyclic group is represented by the following chemical formula U):

=〇 (I) 其中I係擇自氫、鹵素、腈基、醯胺基及硝基所組成之族 群；X係為一碳數1〜5之烷基、烯基或炔基；以及Het係為一 有取代或無取代之雜環基。=〇(I) wherein I is selected from the group consisting of hydrogen, halogen, nitrile, guanamine, and nitro; X is an alkyl, alkenyl or alkynyl group having 1 to 5 carbon atoms; and Het It is a substituted or unsubstituted heterocyclic group.

Het係為一有取代或無取代之雜環基，可舉出之具體例子 如下，但不限於此： 0296-A2060丌 WF(N2);david 1267376Het is a substituted or unsubstituted heterocyclic group, and specific examples thereof are as follows, but are not limited thereto: 0296-A2060丌 WF(N2); david 1267376

上述化學式（i)之製備方法如反應式（1)所示。 反應式（1):The preparation method of the above chemical formula (i) is as shown in the reaction formula (1). Reaction formula (1):

此反應式（1)中，Ri、X及Het範圍均已作如上的詳細界定， 而L-X-Het中的L係為一般用於合成反應中的離去基（leaving group) ° 合成該吲哚醌化合物（I)的反應步驟及條件如下：首先，取 吲哚醌化合物（II)置入例如為無水二曱基曱醯胺（DMF)或二甲基 亞楓(DMSO)的溶劑中，在攝氏-5〜25度下加入例如為氫化鈉 (NaH)或特丁氧基鉀的鹼，在室溫下攪拌反應約0.5〜1.5小時， 接著，取如上述定義的L-X-Het化合物加入上述溶液中，在室 溫下攪拌10〜24小時，之後，加水終止溶液中的反應，接著， 利用有機溶劑如乙醇或二氯甲烷（CH2C12)萃取溶液，待有機層乾 燥、濃縮後，粗產物再以管柱純化之，即可獲得本發明所需的 口弓丨口朵醒產物（I)。 本發明的範圍尚包括化學式（I)對應的鹽類，例如金屬鹽、 胺鹽、酸鹽或有機鹽，形成上述鹽類的方式如后：將化合物（I) 0296-A20607TWF(N2)；david 9 1267376 與等當量的共軛酸或共軛鹼置於醇類溶劑中加熱，再以習知方 法單離所產生的鹽類。 本發明另提供一種包含該吲哚醌化合物或其鹽類的醫藥組 合物，該吲哚醌化合物或其鹽類的有效劑量大約佔該組合物總重 的1〜95%。該組合物更包含藥學上可接受的賦形劑（excipient)、 稀釋劑（diluent)或載體（carrier)，其中所使用的賦形劑包括醣 類、醇類或醣醇類物質、稀釋劑包括澱粉或糊精、載體包括如 固態型式的微脂粒或如液態形式的水溶液、油溶液、水性或油 性懸浮液、酒精溶液或其類似物。本發明醫藥組合物包括口服、 肌肉内或皮下注射等的劑型可由任何適當方法製作而得，其進 入人體後，導致目標區域，也就是感染人體的SARS冠狀病毒 主要蛋白水解酵素受到抑制。 以下藉由數個實施例以更進一步說明本發明之方法、特徵 及優點，但並非用來限制本發明之範圍，本發明之範圍應以所 附之申請專利範圍為準。 【實施例】 製備例1 化合物（1)的合成：In the reaction formula (1), the ranges of Ri, X and Het have been defined as above, and the L system in LX-Het is a leaving group generally used in a synthesis reaction. The reaction step and conditions of the ruthenium compound (I) are as follows: First, the ruthenium compound (II) is placed in a solvent such as anhydrous dimethyl decylamine (DMF) or dimethyl sulfoxide (DMSO). Adding a base such as sodium hydride (NaH) or potassium t-butoxide at -5 to 25 ° C, stirring the reaction at room temperature for about 0.5 to 1.5 hours, and then adding the LX-Het compound as defined above to the above solution. The mixture is stirred at room temperature for 10 to 24 hours. Thereafter, the reaction in the solution is terminated by adding water. Then, the solution is extracted with an organic solvent such as ethanol or dichloromethane (CH 2 C 12 ). After the organic layer is dried and concentrated, the crude product is further The tube string is purified to obtain the desired product (I) of the mouth bow mouth rinse required by the present invention. The scope of the present invention also includes salts corresponding to the chemical formula (I), such as metal salts, amine salts, acid salts or organic salts, and the manner of forming the above salts is as follows: compound (I) 0296-A20607TWF (N2); david 9 1267376 is heated in an alcohol solvent with an equivalent amount of a conjugate acid or a conjugate base, and the resulting salt is isolated by a conventional method. The present invention further provides a pharmaceutical composition comprising the guanidine compound or a salt thereof, wherein the guanidine compound or a salt thereof has an effective dose of about 1 to 95% by weight based on the total weight of the composition. The composition further comprises a pharmaceutically acceptable excipient, diluent or carrier, wherein the excipients used include sugars, alcohols or sugar alcohols, diluents including The starch or dextrin, carrier comprises, for example, a solid type of aliquot or an aqueous solution such as a liquid form, an oil solution, an aqueous or oily suspension, an alcohol solution or the like. The pharmaceutical composition of the present invention comprising a dosage form for oral, intramuscular or subcutaneous injection can be produced by any appropriate method, and after entering the human body, the target region, that is, the main proteolytic enzyme of the SARS coronavirus infected with the human body is inhibited. The method, features and advantages of the present invention are further illustrated by the following examples, which are not intended to limit the scope of the invention, and the scope of the invention should be determined by the scope of the appended claims. [Examples] Preparation Example 1 Synthesis of Compound (1):

合成方法如下： 0296-A20607TWF(N2)；david 10 1267376 1. 取0·5克的5-蛾D引D朵醒(5-iodoisatin)置入6毫升的無水二曱基 甲醯胺中，在攝氏0度下加入60%，0.088克的氫化鈉，於室溫 下攪拌1小時。 、 2. 取0.3 毫升（d=1.17)的 4-(氯曱基）-3,5-二甲基異唑 (4-(chloromethyl)-3,5-dimethylisoxazole)加入步驟 1 溶液中，於 室溫下反應整夜。 3 ·將步驟2溶液倒入冰水中，待沉澱析出後過濾，析出固體用 水及異丙醇沖洗並抽乾，即得0.545克的化合物（1)產物，產率 為 78% 〇 光譜資料： lR NMR(CDC13) δ2.21(3Η) 2.43(3Η) 4·66(2Η) 6.52(1Η) 7.85(1Η) 7·91(1Η) 製備例2 化合物（2)的合成：The synthesis method is as follows: 0296-A20607TWF(N2);david 10 1267376 1. Take 0.5 gram of 5-tox D-dox (5-iodoisatin) and put it into 6 ml of anhydrous dimethylformamide. 60%, 0.088 g of sodium hydride was added at 0 ° C and stirred at room temperature for 1 hour. 2. Add 0.3 ml (d=1.17) of 4-(chloromethyl)-3,5-dimethylisoxazole to the solution in step 1, in the chamber. Warm down all night. 3) The solution of step 2 is poured into ice water, and after precipitation, it is filtered, and the precipitated solid is washed with water and isopropyl alcohol and drained to obtain 0.545 g of the compound (1), the yield is 78%. 〇 Spectral data: lR NMR (CDC13) δ2.21 (3Η) 2.43 (3Η) 4·66(2Η) 6.52(1Η) 7.85(1Η) 7·91(1Η) Preparation 2 Synthesis of Compound (2):

合成方法如下： 0296-A2060丌 WF(N2);david 11 1267376 1·取350毫克的化合物1與16.6毫克的對-甲苯磺酸(p-toluene sulfonic acid)置入15毫升的無水甲醇中，並加入3.06毫升的三 甲基鄰曱酸（trimethyl orthoformate)於攝氏80度下迴流44小時。 2.濃縮去除溶劑後，加入二氯曱烷（CH2C12)及飽和碳酸氫鈉 (NaHC03(sat))進行萃取，有機層經乾燥、濃縮後，粗產物再經 管柱純化（EA/Hex=l/3)，即得3.17毫克的化合物（2)，產率為 81%。 光譜資料： lU NMR(CDC13) δ2.19(3Η) 2.41(3Η) 3_56(6Η) 4.95(2Η) 6·39(1Η) 7·61(1Η) 製備例3 化合物（3)的合成：The synthesis method is as follows: 0296-A2060丌WF(N2);david 11 1267376 1· Take 350 mg of compound 1 and 16.6 mg of p-toluene sulfonic acid in 15 ml of anhydrous methanol, and 3.06 ml of trimethyl orthoformate was added and refluxed at 80 ° C for 44 hours. 2. After removing the solvent by concentration, dichloromethane (CH2C12) and saturated sodium hydrogencarbonate (NaHC03 (sat)) are added for extraction. The organic layer is dried and concentrated, and then the crude product is purified by column (EA/Hex=l/ 3), 3.17 mg of the compound (2) was obtained in a yield of 81%. Spectroscopic data: lU NMR (CDC13) δ 2.19 (3 Η) 2.41 (3 Η) 3_56 (6 Η) 4.95 (2 Η) 6·39 (1 Η) 7·61 (1 Η) Preparation Example 3 Synthesis of Compound (3):

化合物（3) 合成方法如下： 1.取269毫克的化合物2置入6毫升的無水四氫呋喃（丁1^)中， 並加入82·8毫克的氰酸鉀（KCN)與13.5毫克的四（三苯基磷）鈀 0296-A20607TWF(N2)；david 12 1267376 (Tetrakis(triphenylphosphine)palladium)迴流 21 小時。 2.濃縮去除溶劑後，加入二氯曱烷及水進行萃取，有機層經乾 燥、濃縮後，粗產物再經管柱純化（EA/Hex=l/3)，即得201.2毫 克的化合物（3)，產率為98%。 光譜資料： lR NMR(CDC13) δ2.19(3Η) 2.43(3Η) 3.59(6Η) 4.64(2Η) 6·69(1Η) 7.60(1Η) 7·67(1Η) 製備例4 化合物（4)的合成：The synthesis method of the compound (3) is as follows: 1. 269 mg of the compound 2 is placed in 6 ml of anhydrous tetrahydrofuran (butyl 1 ), and 82. 8 mg of potassium cyanate (KCN) and 13.5 mg of tetra (three) are added. Phenylphosphine)palladium 0296-A20607TWF (N2); david 12 1267376 (Tetrakis (triphenylphosphine) palladium) was refluxed for 21 hours. 2. After removing the solvent by concentration, dichloromethane and water are added for extraction, and the organic layer is dried and concentrated, and the crude product is purified by column (EA/Hex=l/3) to obtain 201.2 mg of the compound (3). The yield was 98%. Spectroscopic data: lR NMR(CDC13) δ2.19(3Η) 2.43(3Η) 3.59(6Η) 4.64(2Η) 6·69(1Η) 7.60(1Η) 7·67(1Η) Preparation Example 4 Compound (4) synthesis:

化合物（4) 合成方法如下： 1. 取134毫克的化合物3置入15毫升的乙醇中，於室溫下加 入3Ν，1_5毫升的碳酸納（Na2C03)與30%，1.3毫升的水反應4 小時。 2. 加水稀釋，再以乙醇萃取，待乙醇層乾燥、濃縮後，即得103 毫克的化合物（4)，產率為72.8%。 0296-A20607TWF(N2)；david 13 1267376 光譜資料： !H NMR(CDC13) δ2.20(3Η) 2·42(3Η) 3.60(6Η) 4·62(2Η) 6·67(1Η) 7·78(1Η) 7.90(1Η) 實施例1 化合物（5)的合成：The synthesis of the compound (4) is as follows: 1. 134 mg of the compound 3 is placed in 15 ml of ethanol, 3 Torr is added at room temperature, and 1 to 5 ml of sodium carbonate (Na 2 CO 3 ) is reacted with 30%, 1.3 ml of water for 4 hours. . 2. Dilute with water and extract with ethanol. After the ethanol layer is dried and concentrated, 103 mg of compound (4) is obtained in a yield of 72.8%. 0296-A20607TWF(N2);david 13 1267376 Spectral data: !H NMR(CDC13) δ2.20(3Η) 2·42(3Η) 3.60(6Η) 4·62(2Η) 6·67(1Η) 7·78 (1Η) 7.90 (1Η) Example 1 Synthesis of Compound (5):

化合物（5) 合成方法如下： 1.取53.8毫克的化合物3置入10毫升曱醇與3毫升水的混合 溶劑中，水浴下滴入濃硫酸約28滴。 2·於攝氏70〜75度下反應7小時後，降至室溫並加入飽和碳酸 氫納（NaHC03(sat))調整溶液至中性，續以二氯甲烷萃取，待有 機層乾燥、濃縮後，粗產物再經管柱純化（EA/Hex=l/1)，即得 30毫克的化合物（5)，產率為64%。 光譜資料： NMR(DMSO) δ2.16(3Η) 2.47(3Η) 4.71(2Η) 7.33(1Η) 8.25(1Η) 8·09(1Η) 0296-Α2060丌 WF(N2);david 14 1267376 實施例2 化合物（6)的合成：The synthesis of the compound (5) was as follows: 1. 53.8 mg of the compound 3 was placed in a mixed solvent of 10 ml of decyl alcohol and 3 ml of water, and about 28 drops of concentrated sulfuric acid was added dropwise to the water bath. 2. After reacting for 7 hours at 70 to 75 ° C, the temperature was lowered to room temperature and saturated sodium bicarbonate (NaHC03 (sat)) was added to adjust the solution to neutrality, and extracted with dichloromethane. After the organic layer was dried and concentrated. The crude product was purified by column column (EA/Hex = 1 / 1) to give 30 mg of compound (5) in a yield of 64%. Spectroscopic data: NMR (DMSO) δ 2.16 (3 Η) 2.47 (3 Η) 4.71 (2 Η) 7.33 (1 Η) 8.25 (1 Η) 8·09 (1 Η) 0296-Α2060 丌 WF (N2); david 14 1267376 Example 2 Synthesis of Compound (6):

合成方法如下： 1. 取103毫克的化合物4置入19.5毫升甲醇與6.5毫升水的混 合溶劑中，水浴下滴入濃硫酸約52滴，於攝氏70〜75度下反應 7小時。 2. 濃縮去除一半溶劑後，出現橘色沉澱並過濾。 3. 粗產物經管柱純化，即得30.5克的化合物（6)，產率為34·2%。 光譜資料： lR NMR(DMSO) δ2·16(3Η) 2.47(3Η) 4.71(2Η) 7·21(1Η) 7.40(1Η) 8·03(1Η) 8.07(1Η) 8·17(1Η) 實施例3 化合物（7)的合成： 0296-Α2060丌 WF(N2);david 15 1267376The synthesis was as follows: 1. 103 mg of the compound 4 was placed in a mixed solvent of 19.5 ml of methanol and 6.5 ml of water, and about 52 drops of concentrated sulfuric acid was added dropwise to the water bath, and the reaction was carried out at 70 to 75 ° C for 7 hours. 2. After removing half of the solvent by concentration, an orange precipitate appears and is filtered. 3. The crude product was purified through a column to give 30.5 g of Compound (6). Spectral data: lR NMR (DMSO) δ2·16(3Η) 2.47(3Η) 4.71(2Η) 7·21(1Η) 7.40(1Η) 8·03(1Η) 8.07(1Η) 8·17(1Η) Example 3 Synthesis of compound (7): 0296-Α2060丌WF(N2);david 15 1267376

合成方法如下： 1. 取0.3克的5-碘吲哚醌（5-iodoisatin)置入3毫升的無水二甲 基甲醯胺中，在攝氏0度下加入60%，0·053克的氫化納，於室 溫下攪拌1小時。 2. 取 0.323 克的 2-( 溴甲基）-苯噻吩 (2-(bromomethyl)-benzothiophene)加入步驟 1 溶液中，於室溫下 反應整夜。 3·將步驟2溶液倒入冰水中，待沉澱析出後過濾、，粗產物經管 柱純化，即得0.138克的化合物（7)產物，產率為30%。 光譜資料： NMR(CDC13) δ5.15(2Η) 6·76(1Η) 7.32(3Η) 7·70(1Η) 7_75(1Η) 7_81(1Η) 7.90(1Η) 實施例4 化合物（8)的合成： 0296-A20607TWF(N2);david 16 1267376The synthesis method is as follows: 1. 0.3 g of 5-iodoisatin was placed in 3 ml of anhydrous dimethylformamide, and 60%, 0. 053 g of hydrogenation was added at 0 °C. The mixture was stirred at room temperature for 1 hour. 2. 0.323 g of 2-(bromomethyl)-benzothiophene was added to the solution of step 1, and allowed to react at room temperature overnight. 3. The step 2 solution was poured into ice water, and the precipitate was precipitated and filtered, and the crude product was purified through a column to obtain 0.138 g of the compound (7), yield 30%. Spectroscopic data: NMR (CDC13) δ5.15(2Η) 6·76(1Η) 7.32(3Η) 7·70(1Η) 7_75(1Η) 7_81(1Η) 7.90(1Η) Example 4 Synthesis of Compound (8) : 0296-A20607TWF(N2);david 16 1267376

合成方法如下： 1. 取0.3克的5-碘吲哚醌（5-iodoisatin)置入3毫升的無水二甲 基曱醯胺中，在攝氏0度下加入60%，0.0528克的氫化納，於 室溫下反應1小時。 2. 取 0.327 克的 2-(溴曱基）-1，4-苯二啞烷 (2-(bromomethyl)-l，4-benzodioxane)加入步驟 1 溶液中，於室溫 下反應整夜。 3. 加水終止步驟2溶液中之反應並利用乙醇萃取，有機層乾燥 後濃縮，粗產物經管柱純化，即得143毫克的化合物（8)產物， 產率為31%。 光譜資料： !H NMR(CDC13) δ3·97(1Η) 4.03(2Η) 4·31(1Η) 4·60(1Η) 6.74(2Η) 6·85(3Η) 7·84(2Η) 實施例5 化合物（9)的合成： 0296-A20607TWF(N2)；david 17 1267376The synthesis was as follows: 1. 0.3 g of 5-iodoisatin was placed in 3 ml of anhydrous dimethyl decylamine, and 60%, 0.0528 g of sodium hydride was added at 0 °C. The reaction was carried out for 1 hour at room temperature. 2. Add 0.327 g of 2-(bromomethyl)-1,4-benzodioxane to the solution in step 1, and react overnight at room temperature. 3. The reaction in the solution of step 2 was terminated by adding water and extracted with ethanol. The organic layer was dried and concentrated. The crude product was purified by column column to yield 143 mg of compound (8), yield 31%. Spectral data: !H NMR(CDC13) δ3·97(1Η) 4.03(2Η) 4·31(1Η) 4·60(1Η) 6.74(2Η) 6·85(3Η) 7·84(2Η) Example 5 Synthesis of Compound (9): 0296-A20607TWF (N2); david 17 1267376

化合物（9) 合成方法如下： 1. 取0.3克的5-蛾D弓丨〇朵酉昆（5-iodoisatin)置入3毫升的無水二甲 基甲醯胺中，在攝氏0度下加入60%，0.0528克的氫化納，於 室溫下反應1小時。 2. 取 0.361 克的 2-(溴丙烯基）-苯噻吩 (2-(bromopropenyl)-benzothiophene)加入步驟 1 溶液中，於室溫 下反應整夜。 3·加水終止步驟2溶液中之反應並利用乙醇萃取，有機層乾燥 後濃縮，粗產物經管柱純化，即得342毫克的化合物（91產物， 產率為70% 〇 光譜資料： lR NMR(CDC13) δ4·41(1Η) 6·04(1Η) 6·75(1Η) 6.84(1Η) 7·15(2Η) 7.30(2Η) 7.66(2Η) 7.86(2Η) 實施例ό 化合物（10)的合成： 0296-Α2060丌 WF(N2) ;david 18 1267376 οThe synthesis of the compound (9) is as follows: 1. 0.3 g of 5-odoisatin is placed in 3 ml of anhydrous dimethylformamide and added at 0 °C. %, 0.0528 g of sodium hydride was reacted at room temperature for 1 hour. 2. 0.361 g of 2-(bromopropenyl)-benzothiophene was added to the solution of step 1, and allowed to react at room temperature overnight. 3. Add water to terminate the reaction in step 2 and extract with ethanol. The organic layer is dried and concentrated. The crude product is purified by column to give 342 mg of compound (91 product, yield 70% 〇 spectral data: lR NMR (CDC13 Δ4·41(1Η) 6·04(1Η) 6·75(1Η) 6.84(1Η) 7·15(2Η) 7.30(2Η) 7.66(2Η) 7.86(2Η) Example 合成 Synthesis of compound (10) : 0296-Α2060丌WF(N2) ;david 18 1267376 ο

化合物（10) 合成方法如下： 1·取0·184克的7-璃基D弓丨D朵|昆(7-nitroisatin)置入2毫升的無水 二曱基甲醯胺中，在攝氏0度下加入60%，0.033克的氫化納， 於室溫下反應1小時。 2. 取 0·281 克 的 2塞(溴 甲 基）-苯噻吩 (2-(bromomethyl)-benzothiophene)加入步驟 1 溶液中，於室溫下 反應整夜。 3. 加水終止步驟2溶液中之反應並利用乙醇萃取，有機層乾燥 後濃縮，粗產物經管柱純化，即得177毫克的化合物（10)產物， 產率為50%。 光譜資料： lR NMR(CDC13) δ5.50(2Η) 7·10(1Η) 7.22(1Η) 7·28(2Η) 7·69(2Η) 7.87(2Η) 實施例7 化合物（11)的合成： 0296- A20607TWF(N2);david 19 1267376The synthesis method of the compound (10) is as follows: 1. Take 0. 184 g of 7-glass-based D-pins | 7-nitroisatin is placed in 2 ml of anhydrous dimercaptocaramine at 0 degree Celsius 60%, 0.033 g of sodium hydride was added thereto, and the mixture was reacted at room temperature for 1 hour. 2. Add 0.281 g of 2-(bromomethyl)-benzothiophene to the solution in step 1, and react overnight at room temperature. 3. Add water to terminate the reaction in the solution of step 2 and extract with ethanol. The organic layer is dried and concentrated. The crude product is purified by column to yield 177 mg of compound (10), yield 50%. Spectroscopic data: lR NMR (CDC13) δ5.50 (2Η) 7·10(1Η) 7.22(1Η) 7·28(2Η) 7·69(2Η) 7.87(2Η) Example 7 Synthesis of Compound (11): 0296- A20607TWF(N2);david 19 1267376

合成方法如下： 1 ·取0.108克的7-溴d弓丨D朵醒(7-bromoisatin)置入2毫升的無水二 甲基甲醯胺中，在攝氏0度下加入60%，0.017克的氫化納，於 室溫下攪拌1小時。 2. 取 0.140 克的 2_(溴 曱基）-苯噻吩 (2-(bromomethyl)-benzothiophene)加入步驟 1 溶液中，於室溫下 反應整夜。 3·加水終止步驟2溶液中之反應並利用乙醇萃取，有機層乾燥 後濃縮，粗產物經管柱純化，即得53毫克的化合物（11)產物， 產率為30%。 光譜資料： !H NMR(CDC13) δ5·60(2Η) 6.94(2Η) 7.21(3Η) 7.54(1Η) 7.61(2Η) 7·67(1Η) 實施例8 化合物（12)的合成： 0296-A20607TWF(N2) ；david 20 1267376 οThe synthesis method is as follows: 1) Take 0.108 g of 7-bromoisatin and put it into 2 ml of anhydrous dimethylformamide, and add 60%, 0.017 g at 0 °C. The mixture was stirred at room temperature for 1 hour. 2. 0.140 g of 2-(bromomethyl)-benzothiophene was added to the solution of step 1, and allowed to react at room temperature overnight. 3. The reaction in the solution of step 2 was terminated by adding water and extracted with ethanol. The organic layer was dried and concentrated. The crude product was purified by column column to give 53 mg of compound (11), yield 30%. Spectral data: !H NMR(CDC13) δ5·60(2Η) 6.94(2Η) 7.21(3Η) 7.54(1Η) 7.61(2Η) 7·67(1Η) Example 8 Synthesis of Compound (12): 0296-A20607TWF (N2) ;david 20 1267376 ο

化合物（12) 合成方法如下： 1_取79毫克的5-蛾吲哚醌(5-i〇doisatin)置入1毫升的無水二甲 基甲醯胺中，在攝氏〇度下加入60%，14毫克的氫化納，於室 溫下攪拌1小時。 2· 取 95 毫克的 5-溴曱基-噻吩-2-甲酸四醯胺 (5-bromomethyl-thiophene-2-carbonylic acid butylamide)加入步 驟1溶液中，於室溫下反應整夜。 3.加水終止步驟2溶液中之反應並利用二氯曱烧萃取，有機層 乾燥後濃縮，粗產物經管柱純化，即得11毫克的化合物（12)產 物，產率為8.1% 〇 光譜資料： NMR(DMSO) δ0.89(3Η) 1·24(2Η) 1.50(2Η) 3·20(2Η) 5·07(2Η) 7·15(1Η) 7·22(1Η) 7·56(1Η) 7.84(1Η) 7.85(1Η) 8_45(1Η) 實施例9 化合物（13)的合成： 0296-A20607TWF(N2)；david 21 1267376The synthesis method of the compound (12) is as follows: 1_ Take 79 mg of 5-hypodosin (5-i〇doisatin) and put it into 1 ml of anhydrous dimethylformamide, and add 60% under Celsius. 14 mg of sodium hydride was stirred at room temperature for 1 hour. 2. Add 95 mg of 5-bromomethyl-thiophene-2-carbonylic acid butylamide to the solution of step 1, and react overnight at room temperature. 3. Add water to terminate the reaction in the solution of step 2 and extract with dichlorohydrazine. The organic layer is dried and concentrated. The crude product is purified by column column to obtain 11 mg of compound (12). The yield is 8.1%. NMR (DMSO) δ0.89(3Η) 1·24(2Η) 1.50(2Η) 3·20(2Η) 5·07(2Η) 7·15(1Η) 7·22(1Η) 7·56(1Η) 7.84(1Η) 7.85(1Η) 8_45(1Η) Example 9 Synthesis of Compound (13): 0296-A20607TWF(N2);david 21 1267376

化合物（13) 合成方法如下： 1.取58.4毫克的5-峨D引D朵醒(5-iodoisatin)置入1毫升的無水二 曱基甲醯胺中，在攝氏0度下加入60%，11毫克的氫化納，於 室溫下攪拌1小時。 2·取85毫克的5-溴甲基-噻吩-2-曱酸-4-(氯苯基）醯胺 (5-bromomethyl - thiophene-2-carbonylic acid_4-(chlorophenyl)amide)加入步驟1溶液中，於室溫下反應 整夜。 3.加水終止步驟2溶液中之反應並利用二氯甲烷萃取，有機層 乾燥後濃縮，粗產物經管柱純化，即得7.7毫克的化合物（13) 產物，產率為6.9%。 光譜資料： NMR(DMSO) δ5·07(2Η) 7.15(1Η) 7.22(1Η) 7.39(1Η) 7.41 (2Η) 7.72(2Η) 7·76(2Η) 7·86(1Η) 10.30(1Η) 實施例10 0296-Α2060丌 WF(N2);david 22 1267376 化合物（14)的合成： 〇The synthesis method of the compound (13) is as follows: 1. 58.4 mg of 5-?D D-dodoisatin is placed in 1 ml of anhydrous dimethylformamide, and 60% is added at 0 degree Celsius. 11 mg of sodium hydride was stirred at room temperature for 1 hour. 2. Take 85 mg of 5-bromomethyl-thiophene-2-carbonylic acid_4-(chlorophenyl)amide into the solution of step 1. , react overnight at room temperature. 3. The reaction in the solution of Step 2 was terminated by adding water and extracted with dichloromethane. The organic layer was dried and concentrated. The crude product was purified by column column to yield 7.7 mg of Compound (13). Spectral data: NMR (DMSO) δ5·07(2Η) 7.15(1Η) 7.22(1Η) 7.39(1Η) 7.41 (2Η) 7.72(2Η) 7·76(2Η) 7·86(1Η) 10.30(1Η) Implementation Example 10 0296-Α2060丌WF(N2);david 22 1267376 Synthesis of Compound (14): 〇

化合物（14) 合成方法如下： 1 ·取65毫克的5-蛾D引D朵醒（5-iodoisatin)置入1毫升的無水二甲 基甲醯胺中，在攝氏0度下加入60%，12毫克的氫化納，於室 溫下攪拌1小時。 2. 取104毫克的5-溴甲基-噻吩-2-甲酸-4-(三氟甲基-苯基）醯胺 (5-bromomethyl-thiophene-2-carbonylic acid-4-(trifluoromethyl-phenyl)amide)加入步驟 1 溶液中，於室 溫下反應整夜。 3. 加水終止步驟2溶液之反應並利用二氯甲烷萃取，有機層乾 燥後濃縮，粗產物經管柱純化，即得6.2亳克的化合物Π4)產物， 產率為4.7%。 光譜資料： !H NMR(DMSO) δ5.14(2Η) 7·05(1Η) 7·31(1Η) 7·44(1Η) 7.59(1Η) 7·86(2Η) 7.98(2Η) 8·13(1Η) 10·47(1Η) 0296-Α2060丌 WF(N2) ;david 23 1267376 實施例11 抑制酵素活性測試 EDANS(5-(25aminoethyl)aminonaphthlene sulfonic acid)的 螢 光 發 光 波 長 與 Dabcyl (4-(45-dimethyl-aminobenzeneazo)benzoic acid)的吸收波長是相 互重疊的，因此，EDANS的榮光會因共振能（代80皿110661161^7) 作用而轉移至非勞光（non-fluorescence)的Dabcyl上，使該螢光 發光效果被抑制。利用此原理，將一段SARS proteinase(3CLpro) 的 N 端及 C 端分別接上 Dabcyl 與 EDANS，例 如”Dabcyl-KTSAVLQSGFRKME-EDANS”，貝1 當 3CLpro 切斷此 段胜肽時，偵測到的螢光強度便會隨此段胜肽被3CLpro切斷的 多募而變化（可利用 330/80nm(excitation)及 515/30nm(emission) 偵測螢光強度（RFU))，因而測得酵素活性，此時，若加入不同 濃度的3CLpro抑制劑，即可進一步藉觀察螢光強度變弱的情 形，獲得抑制劑抑制蛋白質的效果。 為了解哪一個抑制劑在其最低濃度時具有較佳抑制 3CLpro活性的效果。首先，測試所有抑制劑，以最終濃度20μΜ 先做篩選，取得抑制效果>50%的抑制劑，再進行不同濃度試 驗，測得抑制劑IC50的濃度。試驗的控制組成份包括10mM的 Tris(reactive buffer)、50nM 的 3CLpro、6μΜ 的胜肽、2.5〇/〇的 DMSO以及稀釋過的Η20，總體積為ΙΟΟμί。試驗組成份則包 括 10mM 的 Tris(reactive buffer)、50ηΜ 的 3CLpro、6μΜ 的胜 肽、20μΜ的抑制劑以及稀釋過的Η20，總體積為lOOgL，每2 分鐘測一次，共10次。所得結果如表一。 0296-A20607TWF(N2) ；david 24 1267376 化合物編號 IC50 化合物編號 ............. IC50 化合物（5) 7.2μΜ 化合物（8) ------ 13·5μΜ 化合物（6) 59·1μΜ 化合物（11) 〇·98μΜ 化合物（7) 0·95μΜ 化合物（14) 12·57μΐνΐ 表一 由表一可看出，本發明化合物（7)抑制酵素活性的效果為最 佳，其在極低濃度例如0·95μΜ的情況下，即可有效抑制3CLpr〇 的活性。 實施例12 專一性試驗 由於人體内有多種酵素，為了確定化合物（7)此一抑制劑具 有專一選擇性，只會抑制3CLpro活性而不會抑制其他酵素的活 性，遂挑選chymotrypsin、trypsin以及papain作為化合物（7)是否 具有選擇性抑制的試驗對照。 首先，進行化合物（7)抑制chymotrypsin活性的試驗，試驗 組成份包含1〇111]\1的丁118〇^3(：1^6 131^6]：)、511]\4的(：]1}011〇1^卩8^1、 200μΜ的胜肽以及濃度分別為ImM、500μΜ、250μΜ、125μΜ、 62·5μΜ與31·25μΜ的化合物（7)，總體積為lOOpL，pH值控制在 8_0。比較試驗結果發現，化合物（7)對3CLpro的抑制效果 (IC50=1 μΜ)較 chymotrypsin(IC50=lmM)明顯為佳，約 500〜1000 倍0 相同地，進行化合物（7)抑制trypsin活性的試驗，試驗組成 份包含 10mM的 Tris(reactive buffer)、9nM的 trypsin、200μΜ的胜 0296-A20607TWF(N2)；david 25 1267376 肽以及濃度分別為 ImM、500μΜ、250μΜ、125μΜ、62·5μΜ與 31·25μΜ的化合物（7)，總體積為100|lil，pH值控制在8.0。比較 試驗結果發現，化合物（7)對3CLpro的抑制效果（Ι〇50=1μΜ)仍較 trypsin(IC50=243pM)為佳，約243倍。 最後，進行化合物（7)抑制papain活性的試驗，試驗組成份 包含5〇111]^的11148〇^&(^^^131^61〇、；13.311]\4的卩3?&^1、1(^1^的胜 肽以及濃度分別為 ImM、500μΜ、250μΜ、125μΜ、62·5μΜ與 31.25μΜ的化合物（7)，總體積為lOOpL，pH值控制在6.2。比較 試驗結果發現，化合物（7)對3CLpro的抑制效果（zcsOMpM)亦較 papain(IC50=87.24pM)為佳，約 87倍之多。 因此，由上述試驗可知，化合物（7)對3CLpro有極專一的抑 制效果，進入人體時不會對體内其他酵素例如chymotrypsin、 trypsin以或papain產生不期望的抑制作用，實可發展成為一高特 異性、有效的抗SARA冠狀病毒藥物。 雖然本發明已以較佳實施例揭露如上，然其並非用以限制 本發明，任何熟習此項技藝者’在不脫離本發明之精神和範圍 内，當可做更動與潤飾，因此本發明之保護範圍當以後附之申 請專利範圍所界定者為準。 0296- A20607TWF(N2);david 26 1267376 【圖式簡單說明】 無。 【主要元件符號說明】 無0 0296-A20607TWF(N2) ；davidThe synthesis method of the compound (14) is as follows: 1) Take 65 mg of 5-iodoisatin and put it into 1 ml of anhydrous dimethylformamide, and add 60% at 0 degree Celsius. 12 mg of sodium hydride was stirred at room temperature for 1 hour. 2. Take 104 mg of 5-bromomethyl-thiophene-2-carbonylic acid-4-(trifluoromethyl-phenyl) The amide) is added to the solution of step 1 and allowed to react overnight at room temperature. 3. Add water to terminate the reaction of step 2 and extract with dichloromethane. The organic layer is dried and concentrated. The crude product is purified by column to give 6.2 g of compound Π4). Spectral data: !H NMR (DMSO) δ5.14(2Η) 7·05(1Η) 7·31(1Η) 7·44(1Η) 7.59(1Η) 7·86(2Η) 7.98(2Η) 8·13 (1Η) 10·47(1Η) 0296-Α2060丌WF(N2) ;david 23 1267376 Example 11 Inhibition of Enzyme Activity Test The fluorescence emission wavelength of EDANS (5-(25aminoethyl)aminonaphthlene sulfonic acid) and Dabcyl (4-( The absorption wavelengths of 45-dimethyl-aminobenzeneazo)benzoic acid are superimposed on each other. Therefore, the glory of EDANS will be transferred to non-fluorescence Dabcyl due to the resonance energy (80 dish 110661161^7). This fluorescent light-emitting effect is suppressed. Using this principle, the N-terminus and C-terminus of a stretch of SARS proteinase (3CLpro) are respectively ligated with Dabcyl and EDANS, for example, "Dabcyl-KTSAVLQSGFRKME-EDANS", and the detected firefly is detected when 3CLpro cuts the peptide. The intensity of the light changes with the multi-raised cut of the peptide by 3CLpro (the fluorescence intensity (RFU) can be detected by using 330/80 nm (excitation) and 515/30 nm (emission)), thereby measuring the activity of the enzyme. At this time, if different concentrations of the 3CLpro inhibitor are added, it is possible to further observe the effect that the inhibitor inhibits the protein by observing the weakening of the fluorescence intensity. To understand which inhibitor has the effect of inhibiting 3CLpro activity at its lowest concentration. First, all inhibitors were tested and screened at a final concentration of 20 μM to obtain an inhibitor of 50% inhibition, and then subjected to different concentration tests to determine the concentration of the inhibitor IC50. The control components of the assay included 10 mM Tris (reactive buffer), 50 nM of 3CLpro, 6 μM peptide, 2.5 〇/〇 DMSO, and diluted Η20 in a total volume of ΙΟΟμί. The test composition consisted of 10 mM Tris (reactive buffer), 50 Μ 3CLpro, 6 μΜ peptide, 20 μM inhibitor, and diluted Η20 in a total volume of 100 gL, measured every 2 minutes for 10 times. The results obtained are shown in Table 1. 0296-A20607TWF(N2) ;david 24 1267376 Compound number IC50 Compound number............. IC50 Compound (5) 7.2μΜ Compound (8) ------ 13·5μΜ Compound ( 6) 59·1μΜ Compound (11) 〇·98μΜ Compound (7) 0·95μΜ Compound (14) 12·57μΐνΐ Table 1 It can be seen from Table 1 that the compound (7) of the present invention has the best effect of inhibiting enzyme activity. It can effectively inhibit the activity of 3CLpr〇 at very low concentrations such as 0·95 μM. Example 12 Specificity test Since there are many enzymes in the human body, in order to determine that the compound (7) has a specific selectivity, it only inhibits the activity of 3CLpro without inhibiting the activity of other enzymes, and selects chymotrypsin, trypsin, and papain as Whether the compound (7) has a test control for selective inhibition. First, a compound (7) was tested for inhibiting the activity of chymotrypsin, and the test composition contained 1〇111]\1 of 〇118〇^3(:1^6 131^6]:), 511]\4 of (:]1 }011〇1^卩8^1, 200μΜ peptide and compound (7) with concentrations of ImM, 500μΜ, 250μΜ, 125μΜ, 62·5μΜ and 31·25μΜ, respectively, the total volume is lOOpL, and the pH is controlled at 8_0. The results of comparative experiments showed that the inhibitory effect of compound (7) on 3CLpro (IC50=1 μΜ) was better than that of chymotrypsin (IC50=lmM), about 500~1000 times 0, and the compound (7) inhibited trypsin activity. The test composition contains 10 mM Tris (reactive buffer), 9 nM trypsin, 200 μΜ of Win 0296-A20607TWF (N2); david 25 1267376 peptide and concentrations of ImM, 500 μΜ, 250 μΜ, 125 μΜ, 62·5 μΜ and 31·25 μΜ, respectively. Compound (7), the total volume is 100|lil, and the pH is controlled at 8.0. Comparing the test results, it is found that the inhibitory effect of compound (7) on 3CLpro (Ι〇50=1μΜ) is still better than trypsin (IC50=243pM). , about 243 times. Finally, the compound (7) inhibits the activity of papain, the test composition胜3?&^1,1(^1^) containing 11〇〇&(^^^131^61〇,;13.311]\4 of 5〇111]^ and the concentration of I peptide, 500μΜ 250μΜ, 125μΜ, 62·5μΜ and 31.25μΜ of compound (7), the total volume is lOOpL, the pH value is controlled at 6.2. The comparison test results show that the inhibitory effect of compound (7) on 3CLpro (zcsOMpM) is also higher than that of papain (IC50). =87.24 pM) is preferably about 87 times. Therefore, it can be seen from the above test that the compound (7) has a very specific inhibitory effect on 3CLpro, and does not affect other enzymes in the body such as chymotrypsin, trypsin or papain when entering the human body. Unexpected inhibition can be developed into a highly specific and effective anti-SARA coronavirus drug. Although the invention has been disclosed in the preferred embodiments as above, it is not intended to limit the invention, and any skill in the art is The scope of protection of the present invention is defined by the scope of the appended claims, without departing from the spirit and scope of the invention. 0296- A20607TWF(N2);david 26 1267376 Simple description of the schema] None. [Main component symbol description] No 0 0296-A20607TWF(N2) ;david

Claims (1)

38582號申請專利範圍修正本 修正日期：95.10.3 十、申請專利範圍： 1.一種叫Ισ朵Sft(isatin)化合物，具有化學式（I)之結構或其鹽 類：Amendment No. 38582 Patent Application Scope Amendment Date: 95.10.3 X. Patent Application Range: 1. A compound called Sft (isatin) having the structure of chemical formula (I) or its salts: 其中係包括腈基或醯胺基；X係為一碳數1〜5之烷基、 烯基或炔基；以及 Het 係包括下列其一：Wherein it includes a nitrile group or a guanamine group; the X system is an alkyl group, an alkenyl group or an alkynyl group having 1 to 5 carbon atoms; and the Het system includes one of the following: 2.如申請專利範圍第1項所述之吲哚醌化合物，其中該化 合物係包括2. The hydrazine compound of claim 1, wherein the compound comprises 0296-A20607TWF2(N2)；david 280296-A20607TWF2(N2);david 28 s•如申請專利範圍第丨項 ,、斤这之°引哚醌化合物，其中該4 :秦“蛋白水解酵素之抑制劑。 •如申請專利範圍第3頊 K CAOO 員所述之吲哚醌化合物，其中冠ί| 門勢係為SARS冠狀病毒。 5·一種醫藥組合物，包括： —如申請專利範圍第丨韻述之㈣K (isatm)化合物幻 鹽類；以及 一樂學上可接受之賦形劑（excipient)、稀釋劑（diluem)或⑽ 體（carrier)，其中該醫藥組合物係為一 SARS冠狀病毒蛋白水角 0296-A2060丌 WF2(N2);david 29 1267376 酵素之抑制劑。 6. 如申請專利範圍第5項所述之醫藥組合物，其中該吲哚 醌化合物或其鹽類之有效劑量大體為1〜95%。 7. 如申請專利範圍第5項所述之醫藥組合物，其中該醫藥 組合物之劑型係包括口服、肌肉内或皮下注射。 0296-A20607TWF2(N2)；david 30s • If you apply for the scope of the patent, the sputum compound, which is the “inhibitor of proteolytic enzymes.” • As described in the third section of the patent application, K CAOO a compound, wherein the gate is a SARS coronavirus. 5. A pharmaceutical composition comprising: - a compound salt of the K (isatm) compound as described in the patent application, and a musically acceptable An excipient, a diluent or a carrier, wherein the pharmaceutical composition is a SARS coronavirus protein water angle 0296-A2060 丌 WF2 (N2); an inhibitor of david 29 1267376 enzyme. 6. The pharmaceutical composition according to claim 5, wherein the effective dose of the bismuth compound or a salt thereof is generally from 1 to 95%. 7. The pharmaceutical composition as described in claim 5 The dosage form of the pharmaceutical composition includes oral, intramuscular or subcutaneous injection. 0296-A20607TWF2(N2);david 30