CN112220808A - 一种麻黄提取物及其活性成分和用途 - Google Patents
一种麻黄提取物及其活性成分和用途 Download PDFInfo
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- CN112220808A CN112220808A CN202011294142.5A CN202011294142A CN112220808A CN 112220808 A CN112220808 A CN 112220808A CN 202011294142 A CN202011294142 A CN 202011294142A CN 112220808 A CN112220808 A CN 112220808A
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- ephedra
- disinfectant
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- novel coronavirus
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Abstract
本发明涉及药学领域，特别是涉及一种可抑制新型冠状病毒3CL蛋白水解酶的麻黄提取物及其活性成分及其用途。所述麻黄提取物为麻黄水提物和麻黄醇提物，所述活性成分包括草棉黄素、没食子酸、槲皮素及芹菜素。本发明的麻黄提取物及其活性成分可用于治疗新型冠状病毒感染。
Description
技术领域
本发明属于药学领域，特别是涉及一种麻黄提取物及其活性成分及其医药用途。
背景技术
由新冠病毒(SARS-CoV-2)引起的病毒性肺炎(COVID-19)已成为国际关注的突发公共卫生事件。该病毒一般通过呼吸道分泌物排出，可通过飞沫传播，也可通过接触传播等。在病毒感染宿主的过程中，新冠病毒中的3CL蛋白水解酶(又称为Mpro)是一类半胱氨酸水解酶，能够在病毒复制过程中裂解多聚蛋白，生成多个有活性的功能蛋白，在病毒的转录和复制中起重要作用。因此，3CL蛋白水解酶可作为抗新冠病毒的关键安全的治疗靶标。目前对于新冠病毒感染的预防和治疗临床尚无特效药。因此，以新冠病毒3CL蛋白水解酶为靶标，寻找一种安全高效，且对3CL蛋白酶水解活性有显著抑制的中草药或其活性部位，将对于新冠病毒的预防及辅助治疗具有重要意义。
麻黄为麻黄科麻黄属草本状灌木草麻黄Ephedra sinica Stapf.、中麻黄Ephedraintermedia Schrenk et C.A.Mey.或木贼麻黄Ephedra equisetina Bge.的干燥草质茎，为临床常用的中药，有疏解机表、化痰止咳、利水消肿等功效，用于风寒感冒、胸闷喘咳、风湿痹痛等症。
发明内容
针对当前抗新冠病毒药物研发现状，本申请的发明人以新冠3CL蛋白水解酶为靶点，开展了中草药及其化学成分抑制新冠3CL蛋白水解酶的规模化筛选，结果发现麻黄水提物、麻黄非生物碱部位及其主要单体成分草棉黄素等对新冠病毒3CL蛋白水解酶水解活性均有显著的抑制作用。故而，可将其应用于药物、口服液、漱口水、消毒湿巾、消毒液及外用洗液中，可用于新冠病毒的消杀和日常预防。
进而，本发明首先提供了一种可抑制新型冠状病毒3CL蛋白水解酶的麻黄水提物，所述麻黄水提物可由如下方法制得：取1kg麻黄，粉碎，加10L的纯水，浸泡30分钟，加热至沸，煎煮20分钟，收集药液；再加8L纯水，加热至沸，煎煮15分钟，合并煎液，过滤，煎液减压浓缩至2L，浓缩液喷雾干燥，收集干粉，即得。
本发明的第二方面是提供了所述的麻黄水提物在制备可抑制新型冠状病毒3CL蛋白水解酶的药物中的应用。
本发明的第三方面是提供了所述的麻黄水提物在制备可抑制新型冠状病毒3CL蛋白水解酶的口服液或消毒洗液制品中的应用，所述消毒洗液制品选自洗手液、消毒液、消毒凝胶、洗衣液、沐浴液、洗发剂、洗洁精、清洁剂、漱口液或消毒湿纸巾。
本发明的第四方面是提供了一种可抑制新型冠状病毒3CL蛋白水解酶的麻黄醇提物，所述麻黄醇提物可由如下方法制得：将麻黄粉碎，按料液比1：5用75％-95％乙醇浸泡，80℃下回流提取3次，过滤得提取液，重复提取三次后合并提取液，加甲醇溶解再用正庚烷萃取3次，回收溶剂，得甲醇层提取物和正庚烷层提取物；取甲醇层提取物溶于体积比为25：75：0.1的甲醇-水-甲酸溶液中，再以12 000r/min离心20min，得上清液和沉淀；取上清液上样AC18 SPE柱，依次用体积比为0.1：100的甲酸-水和体积比为5：95：0.1的甲醇-水-甲酸溶液洗脱，富集，真空减压、浓缩干燥。
较优选的，所述的麻黄醇提物含有草棉黄素、没食子酸、槲皮素和芹菜素。
本发明的第五方面是提供了上述麻黄醇提物在制备可抑制新型冠状病毒3CL蛋白水解酶的药物中的应用。
本发明的第六方面是提供了上述麻黄醇提物在制备可抑制新型冠状病毒3CL蛋白水解酶的口服液或消毒洗液制品中的应用，所述消毒洗液制品选自洗手液、消毒液、消毒凝胶、洗衣液、沐浴液、洗发剂、洗洁精、清洁剂、漱口液或消毒湿纸巾。
本发明的第七方面是提供了草棉黄素或没食子酸或槲皮素或芹菜素在制备可抑制新型冠状病毒3CL蛋白水解酶的药物中的应用。
本发明还提供了草棉黄素或没食子酸或槲皮素或芹菜素在制备可抑制新型冠状病毒3CL蛋白水解酶的口服液或消毒洗液制品中的应用，所述消毒洗液制品选自洗手液、消毒液、消毒凝胶、洗衣液、沐浴液、洗发剂、洗洁精、清洁剂、漱口液或消毒湿纸巾。
附图说明
图1是麻黄生物碱部位与麻黄非生物碱部位抑制新冠3CLpro的初筛结果，表明麻黄非生物碱部位对新冠3CLpro有显著的抑制作用。
图2是麻黄非生物碱部位的单体成分抑制新冠3CLpro的初筛结果，显示出没食子酸，草棉黄素，槲皮素和芹菜素对新冠3CLpro有抑制作用。
图3是实施例四中麻黄水提物和麻黄非生物碱部位对新冠3CLpro抑制曲线，表明麻黄水提物可剂量依赖性地抑制新冠3CLpro活性。
图4是实施例四中麻黄非生物碱部位的活性成分(草棉黄素、没食子酸、槲皮素和芹菜素)对新冠3CLpro抑制曲线，表明麻黄非生物碱可剂量依赖性地抑制新冠3CLpro活性。
图5是实施例五中草棉黄素对新冠3CLpro时间及剂量依赖性抑制曲线，表明草棉黄素可时间及剂量依赖性灭活新冠病毒3CLpro。
图6为是实施例六中草棉黄素对新冠3CLpro抑制的KI曲线，表明草棉黄素可时间及剂量依赖性灭活新冠3CLpro。
具体实施方式
本发明人在前期高通量筛选结果基础上，惊喜地发现麻黄水提物能够明显抑制新冠病毒3CLpro，进一步分离生物碱部位和非生物碱部位，发现麻黄生物碱部位对3CLpro没有抑制作用，而麻黄非生物碱部位对3CLpro有强效抑制作用。因此，结合UHPLC-Q-ExactiveOrbitrap HRMS对麻黄非生物碱部位主要成分进行鉴定，锁定了其活性成分为草棉黄素、没食子酸、槲皮素和芹菜素。分别绘制了麻黄水提物、麻黄非生物碱部位及其活性成分对新冠3CLpro的抑制曲线，拟合了其IC50值，明确其对3CLpro水解活性的强效抑制能力。
本发明所述的麻黄水提物、麻黄非生物碱部位及其活性成分还可做本草精华添加至各种外用洗剂中，相比常用洗涤剂或消毒剂，其具备阻碍新冠病毒复制的活性成分，刺激性小，安全无毒，可在低剂量下高效抑制新冠病毒3CLpro。
下面结合具体实施例，进一步阐述本发明。应理解，这些实施例仅用于说明本发明而不用于限制本发明的范围。下列实施例中未注明具体条件的实验方法，通常按照常规条件或按照制造厂商所建议的条件。除非另外说明，否则所有的百分数、比率、比例、或份数按重量计。
除非另行定义，文中所使用的所有专业与科学用语与本领域熟练人员所熟悉的意义相同。此外，任何与所记载内容相似或均等的方法及材料皆可应用于本发明方法中。文中所述的较佳实施方法与材料仅作示范之用。
本发明提到的上述特征，或实施例提到的特征可以任意组合。本专利说明书所揭示的所有特征可与任何组合物形式并用，说明书中所揭示的各个特征，可以任何可提供相同、均等或相似目的的替代性特征取代。因此除有特别说明，所揭示的特征仅为均等或相似特征的一般性例子。
实施例一麻黄水提物的制备
1.材料
干燥麻黄草质茎；Milli-Q Direct 8纯水/超纯水一体化系统(Bedford，美国)；旋转蒸发仪(东京理化公司)；煎药机与喷雾干燥机(上海姚氏仪器设备)。
2.方法
取1kg麻黄，切碎投入20L的煎药机内，加10L的纯水，浸泡30分钟，加热至沸，煎煮20分钟，收集药液；继加8L纯水，加热至沸，煎煮15分钟，合并煎液，过滤，煎液于20L旋转蒸发仪中在温度50℃、真空度为-0.095MPa的条件下减压浓缩至2L，浓缩液于喷雾干燥机中在进风口温度130℃、出风温度75℃以上以及雾化器压力0.2MPa的条件下进行喷雾干燥，收集干粉，避光密封备用。
实施例二麻黄醇提物的制备
1.材料
干燥麻黄草质茎；甲醇、甲酸为色谱纯，其他试剂均为分析纯。Milli-Q Direct 8纯水/超纯水一体化系统(Bedford，美国)；KQ-800DE型数码超声仪(昆山市超声仪器有限公司)；EYELA旋转蒸发仪(东京理化公司)；低温高速离心机(Thermo)，YHG-9245A型电热恒温鼓风干燥箱(上海姚氏仪器设备)。
2.方法
将麻黄粉碎，按料液比1：5用85％乙醇浸泡，80℃下回流提取3次，过滤得提取液，重复提取三次后合并提取液，加甲醇充分溶解再用正庚烷萃取3次，回收溶剂，得甲醇层提取物和正庚烷层提取物。取甲醇层提取物溶于50mL甲醇-水-甲酸(25：75：0.1，V/V/V)溶液中，再以12 000r/min离心20min，得上清液和沉淀部分。取该上清液上样AC18 SPE柱，依次用10mL甲酸-水(0.1：100，V/V)和50mL甲醇-水-甲酸(5：95：0.1，V/V/V)溶液洗脱，富集，真空减压、浓缩干燥。制得麻黄非生物碱部位干粉，避光密封备用。
实施例三麻黄醇提物主要成分的鉴定
1.材料与方法
1.1材料
麻黄醇提物粉末(按实施例二制备)，UHPLC-Q-Exactive Orbitrap HRMS液质联用系统：Dionex Ultimate 3000高压液相色谱串联Q Exactive四极杆/静电场轨道阱高分辨质谱仪(美国Thermo Fisher Scientific公司)；Acquity
BEH C18色谱柱(100mm×2.1mm，1.7μm，美国Waters公司)；
1.1方法
色谱条件是使用Dionex Ultimate 3000高压液相系统，Acquity 
BEH C18色谱柱(100mm×2.1mm,1.7μm)；流动相为0.1％甲酸水与甲醇，梯度洗脱，洗脱程序为0～2min，4％甲醇；2～6min，4％～12％甲醇；6～38min，12％-70％甲醇；38～38.5min，70％甲醇；38.5～39min，70％～95％甲醇；39～43min，95％甲醇；43～45min，4％甲醇。流速均为0.3mL·min-1；柱温均为40℃，进样量均为2μL。质谱条件如下：使用Q Exactive四极杆-静电场轨道阱高分辨质谱仪作为分析检测的仪器；离子源为电喷雾离子源(H-ESI)，辅助气流量13arb，辅助加热器温度300℃，离子传输管温度320℃，自动增益控制(AGC)106，s-lens射频水平50；正离子模式：鞘气(N2)流量35arb，喷雾电压3.5kV；负离子模式：鞘气(N2)流量35arb，喷雾电压2.5kV。扫描方式采用正负离子Full MS/dd-MS2模式，其中包括1次一级全扫描(分辨率为70000FWHM)和1次数据依赖的二级扫描(分辨率为17500FWHM)2个事件，正负离子扫描范围均为m/z 80～1200，碰撞能梯度为20V、50V、100V。
2.结果
UHPLC-Q-Exactive Orbitrap HRMS对麻黄非生物碱部位主要成分的鉴定，结果如下表所示。
实施例四麻黄水提物、麻黄醇提物及其单体成分抑制新冠3CLpro的IC50测定
1.材料与方法
1.1材料
麻黄水提物干粉，麻黄醇提物按实施例一和二制备。儿茶素，表儿茶素购自于上海诗丹德生物技术有限公司。牡荆素，异牡荆素，小麦黄素，芦丁，草棉黄素，槲皮苷，异槲皮苷，柚皮素，二氢槲皮素，山奈酚，芹菜素，槲皮素，没食子酸购自于成都普菲德生物科技有限公司。新冠3CLpro由上海中医药大学交叉科学研究院葛广波课题组重组表达，作为体外3CLpro抑制实验的酶源；底物Dabcyl-KNSTLQSGLRKE-Edans由Sangon Biotech公司合成(上海)；磷酸二氢钾、磷酸氢二钾、乙二胺四乙酸均购自美仑生物(纯度>98％)；色谱级二甲基亚砜购自Tedia公司(美国)；Milli-Q Direct 8纯水/超纯水一体化系统(Bedford，美国)；2.5μL、10μL、20μL、100μL、200μL、1000μL移液器(德国Eppendorf)；黑色96孔板；涡旋仪；恒温混匀仪(杭州奥盛)；
ID3多模式酶标仪(奥地利)。
1.2方法
向96孔板中加入90μL预孵液，预孵液由pH＝7.4，含1mM乙二胺四乙酸(EDTA)的磷酸盐缓冲液78μL、不同浓度的抑制剂2μL、10μL新冠3CLpro(终浓度为4μg/mL)组成。将预孵液在37℃下预孵60分钟后，加入终浓度为20μM的底物(Dabcyl-KNSTLQSGLRKE-Edans)起始反应，于多功能酶标仪下对底物的代谢水解产物连续检测30分钟(激发波长为340nm，发射波长为490nm)。随后绘制其抑制曲线，由Graph Pad Prism 7.0软件进行数据处理。
其中剩余酶活的计算公式为：
剩余酶活(％)＝(F0-F1)/F0×100％
F0为预孵液中不加抑制剂时测得的荧光强度值，F1为预孵液中加入各浓度抑制剂时测得的荧光强度值。
2.结果
如图1-4所示，麻黄水提物，非生物碱部位及其活性成分能够抑制新冠3CLpro，并且呈剂量依赖型，其IC50值如下表所示。
实施例五草棉黄素呈时间及剂量依赖性抑制新冠3CLpro
1.材料与方法
1.1材料
草棉黄素购自于成都普菲德生物科技有限公司；新冠3CL蛋白酶由上海中医药大学交叉科学研究院葛广波课题组重组表达；底物Dabcyl-KNSTLQSGLRKE-Edans由SangonBiotech公司合成(上海)；磷酸二氢钾、磷酸氢二钾、乙二胺四乙酸均购自美仑生物(纯度>98％)；色谱级二甲基亚砜购自Tedia公司(美国)；Milli-Q Direct 8纯水/超纯水一体化系统(Bedford，美国)；2.5μL、10μL、20μL、100μL、200μL、1000μL移液器(德国Eppendorf)；黑色96孔板；涡旋仪；恒温混匀仪(杭州奥盛)；
ID3多模式酶标仪(奥地利)。
1.2方法
反应体系体积为100μL，首先向96孔板中加入预孵液，预孵液由PBS-EDTA缓冲液(pH 7.4)、终浓度为4μg/mL的新冠3CL蛋白酶和不同浓度的麻黄非生物碱组成。将预孵液在37℃恒温金属浴震荡，分别孵育3min和60min。再向预孵液中加入终浓度为20μM的Dabcyl-KNSTLQSGLRKE-Edans荧光底物，起始反应，接着采用多模式酶标仪，检测水解产物(λex＝340nm，λem＝490nm)在采集波长处的荧光强度值。随后以抑制剂浓度的对数为横坐标，剩余酶活为纵坐标，绘制其抑制曲线，由Graph Pad Prism 7.0软件进行数据处理。
其中剩余酶活的计算公式为：
剩余酶活(％)＝(F0-F1)/F0×100％
F0为预孵液中不加草棉黄素时测得的荧光强度值，F1为预孵液中加入各浓度草棉黄素时测得的荧光强度值。
2.结果
如图5所示，草棉黄素呈时间及剂量依赖性抑制新冠3CLpro，其IC50值如下表：
实施例六草棉黄素灭活新冠3CLpro的灭活常数KI测定
1.材料与方法
1.1材料
草棉黄素购自于成都普菲德生物科技有限公司；SARS-2 3CL蛋白酶及SARS 3CL蛋白酶由上海中医药大学交叉科学研究院葛广波课题组重组表达，作为体外3CL蛋白酶抑制实验的酶源；底物Dabcyl-KNSTLQSGLRKE-Edans由Sangon Biotech公司合成(上海)；磷酸二氢钾、磷酸氢二钾、乙二胺四乙酸均购自美仑生物(纯度>98％)；色谱级二甲基亚砜购自Tedia公司(美国)；Milli-Q Direct 8纯水/超纯水一体化系统(Bedford，美国)；2.5μL、10μL、20μL、100μL、200μL、1000μL移液器(德国Eppendorf)；黑色96孔板；涡旋仪；恒温混匀仪(杭州奥盛)；
ID3多模式酶标仪(奥地利)。
1.2方法
首先准备一系列失活孵育样品A(100μL)和3CL蛋白酶活性评价孵育样品B(90μL)，失活孵育样品A由不同浓度的草棉黄素和3CL蛋白酶组成。样品A按不同时间点在恒温孵育器上37℃分别预孵10，20，30，50，60后，各取出10μL，然后迅速加入3CL蛋白酶活性评价孵育样品B中起始反应，样品B包含80μL PBS-EDTA缓冲液(pH 7.4)及荧光底物Dabcyl-KNSTLQSGLRKE-Edans，反应20min后，加入等体积的冰乙腈终止反应，涡旋混匀。接着从终止反应后的样品B中取出100μL至96孔板中，采用多模式酶标仪，检测水解产物(λex＝340nm，λem＝490nm)在采集波长处的荧光强度值。3CL蛋白酶残余活性的计算公式：残余活性(％)＝(抑制剂存在时测得的荧光强度值)/空白组测得的荧光强度值×100％，绘制抑制曲线，计算KI。
两个样品组：
A.失活孵育样品
B.SARS-2 3CL活性评价孵育样品
2.结果
结果如图6所示，草棉黄素能够呈时间及剂量依赖性强效抑制新冠3CL蛋白酶，其KI值为18.87μM，Kinact为0.01min-1。
实施例七麻黄口服液的制备
配方(质量比)：20份麻黄水提物或其非生物碱部位，4份蜂蜜，0.5份果胶、0.5份山梨酸钾和75份去离子水。将以上组分加入搅拌器中进行混匀调配，之后进行高温杀菌，测定相关技术指标，合格后灌装完成后，进行二次杀菌，制得麻黄饮料或不同包装规格的口服液。
实施例八麻黄精华消毒湿纸巾的制备
配方(质量比)：30份的麻黄水提物或其非生物碱部位，30份的乙醇，0.5份十二烷基二甲基氧化胺、0.5份月桂醇聚氧乙烯醚、2份木糖醇和37份去离子水。将以上组分混匀，通过预浸渍方法喷淋到基材上，经过湿巾机器裁切、测定相关技术指标，合格后灭菌包装，制得麻黄精华消毒湿巾。
实施例九麻黄精华洗手液的制备
配方(质量比)：10份麻黄水提物或其非生物碱部位、10份乙醇、4份烷基糖苷、0.6份乙二胺四乙酸，3份甜菜碱、2份山梨醇、2份柠檬酸、0.4份柠檬酸钠和68份去离子水组成。将溶剂、甘油、氯化钠、柠檬酸钠和柠檬酸等缓慢依次加入到37±3℃的去离子水中。搅拌均匀后静置15分钟后温度降至常温状态时，测定相关技术指标，合格后包装，制得麻黄精华洗手液。
实施例十麻黄精华漱口液的制备
配方(质量比)：20份麻黄水提物或其非生物碱部位、0.1份甘油、5份乙醇、0.5表面活性剂、0.5份辛甘醇、2份山梨醇、0.5份柠檬酸、0.4份防腐剂、0.5份水溶性薄荷香料和70.5份去离子水组成。在有加热装置的反应中加入水，加热至37±3℃，将主要组分依次缓慢加入到热水中，搅拌均匀，加入柠檬酸调节pH值，最后加入水溶性薄荷香料，温度降至常温状态时，检测相关指标，合格后杀菌装瓶，制得麻黄精华漱口液。
实施例十一麻黄精华洗洁剂的制备
配方(质量比)：10份麻黄水提物或其非生物碱部位、8份脂肪醇聚氧乙烯醚硫酸钠，起泡剂：2份椰子油脂肪酸二乙醇酰胺、1份防腐剂、1.3份氢氧化钠、5份脂肪醇聚氧乙烯醚、10份直链烷基苯磺酸、0.1份乙二胺四乙酸二钠、1份氯化钠、0.6香精和61份去离子水组成。将定量的去离子水投入配料锅中，在搅拌下加入氢氧化钠，待其溶解后缓缓加入直链烷基苯磺酸，搅拌调节pH，在温度下加入剩余组分，澄清透明后加入香精，补足去离子水即可。温度降至常温状态时，测定相关技术指标，合格后包装，制得麻黄精华洗洁剂。
实施例十二麻黄精华洗发液的制备
配方(质量比)：20份麻黄水提物或其非生物碱部位、15份表面活性剂、2份珠光浆、3份保湿柔亮剂、2份增稠剂、3份柠檬酸、1份香精、1份色素、1份防腐剂、2份发泡剂和50份去离子水。反应釜中加入麻黄水提物及非生物碱部位、表面活性剂、珠光浆、保湿柔亮剂、增稠剂和去离子水，连续搅拌混匀乳化，加入柠檬酸调节pH，持续真空搅拌25-30分钟；最后加入香精、色素、防腐剂、发泡剂再真空搅拌25-30分钟，测定相关技术指标，合格后包装，制得麻黄精华洗发液。
实施例十三麻黄精华洗衣液的制备
配方(质量比)：50份麻黄水提物或其非生物碱部位，15份椰子油脂肪酸甘氨酸钾、4份柠檬酸、15份纳米二氧化钛、10份氯化十二烷基二甲基苄基铵、3份羧甲基纤维素、3份硅酸钠。将麻黄水提物或其非生物碱部位、椰子油脂肪酸甘氨酸钾、柠檬酸、纳米二氧化钛加入搅拌罐中均匀20分钟后，加入氯化十二烷基二甲基苄基铵、羧甲基纤维素、硅酸钠混合后加入加热罐中低温加热，50℃加热20分钟，之后缓慢冷却至室温，测定相关技术指标，静置5h后，合格后包装，制得麻黄精华洗衣液。
实施例十四麻黄精华消毒液的制备
配方(质量比)：30份麻黄水提物或其非生物碱部位，30份乙醇、3份双氧水、6份甘油、1份乙二胺四乙酸四钠及30份去离子水。将麻黄水提物或其非生物碱部位和乙醇加入搅拌器中搅拌10-20分钟，随后加入去离子水和双氧水，最后加入甘油和乙二胺四乙酸四钠，搅拌25-30分钟使液体混合均匀。检测合格后灭菌包装，制得麻黄精华消毒液。
实施例十五麻黄精华全能清洁剂的制备
配方(质量比)：30份麻黄水提物或其非生物碱部位、20份椰子油脂肪醇单乙醇酰胺、5份柠檬酸、10份烷基酚聚氧乙烯醚、3.8份氢氧化钠、6.2份二甲基苯磺酸钠、5份偏硅酸钠20份去离子水。将麻黄醇提物、椰子油脂肪醇单乙醇酰胺、烷基酚聚氧乙烯醚、偏硅酸钠加入到反应釜中反应20-30分钟，再依次添加其余组分，搅拌均匀至透明，加入柠檬酸调节pH值，再进行自然消泡静置后除去沉淀，测定相关指标，合格后灌装，制得麻黄精华全能清洁剂。
实施例十六麻黄精华沐浴液的制备
配方(质量比)：10份麻黄水提物或其非生物碱部位、50份脂肪醇聚氧乙烯醚硫酸铵、10份月桂醇硫酸钠、5份脂肪酸甲酯磺酸盐、3份乳化硅油、3份椰子油脂肪酸二乙醇酰胺、0.1份水杨酸、2份杏仁油、0.5份柠檬酸和13.4份去离子水组成。将杏仁油和椰子油脂肪酸二乙醇酰胺混合为油相，加热到75℃融化成液体保温备用，其余组分加热后加入，搅拌混合，柠檬酸调节pH，均质成稳定乳液后降温静置消泡，测定相关指标，合格后灌装，制得麻黄精华沐浴液。
实施例十七麻黄精华免洗消毒凝胶的制备
配方(质量比)：30份麻黄水提物或其非生物碱部位、30份乙醇、1份三氯羟基二苯醚、10份卡波姆940、4份山梨醇和25份去离子水。制作工艺：将卡波姆940加入到有水的反应釜中静止溶胀30分钟，制备出A溶液；再将其余组分在另一容器中搅拌均匀，调节pH值，制备出B溶液；再将B溶液加入A溶液中，搅拌均匀灌装，制得麻黄精华免洗消毒凝胶。
本发明所涉及的多个方面已做如上阐述。然而，应理解的是，在不偏离本发明精神之前提下，本领域专业人员可对上述发明进行等同改变、工艺微调和修饰，所述改变、工艺微调和修饰同样落入本专利申请之权利要求的覆盖范围。
Claims (9)
1.一种可抑制新型冠状病毒3CL蛋白水解酶的麻黄水提物，其特征在于，所述麻黄水提物可由如下方法制得：取1kg麻黄，粉碎，加10L的纯水，浸泡30分钟，加热至沸，煎煮20分钟，收集药液；再加8L纯水，加热至沸，煎煮15分钟，合并煎液，过滤，煎液减压浓缩至2L，浓缩液喷雾干燥，收集干粉，即得。
2.如权利要求1所述的麻黄水提物在制备可抑制新型冠状病毒3CL蛋白水解酶的药物中的应用。
3.如权利要求1所述的麻黄水提物在制备可抑制新型冠状病毒3CL蛋白水解酶的口服液或消毒洗液制品中的应用，所述消毒洗液制品选自洗手液、消毒液、消毒凝胶、洗衣液、沐浴液、洗发剂、洗洁精、清洁剂、漱口液或消毒湿纸巾。
4.一种可抑制新型冠状病毒3CL蛋白水解酶的麻黄醇提物，其特征在于，所述麻黄醇提物可由如下方法制得：将麻黄粉碎，按料液比1：5用75％-95％乙醇浸泡，80℃下回流提取3次，过滤得提取液，重复提取三次后合并提取液，加甲醇溶解再用正庚烷萃取3次，回收溶剂，得甲醇层提取物和正庚烷层提取物；取甲醇层提取物溶于体积比为25：75：0.1的甲醇-水-甲酸溶液中，再以12 000r/min离心20min，得上清液和沉淀；取上清液上样AC18 SPE柱，依次用体积比为0.1：100的甲酸-水和体积比为5：95：0.1的甲醇-水-甲酸溶液洗脱，富集，真空减压、浓缩干燥。
5.如权利要求4所述的麻黄醇提物，其特征在于，所述麻黄醇提物含有草棉黄素、没食子酸、槲皮素和芹菜素。
6.如权利要求4或5所述的麻黄醇提物在制备可抑制新型冠状病毒3CL蛋白水解酶的药物中的应用。
7.如权利要求4或5所述的麻黄醇提物在制备可抑制新型冠状病毒3CL蛋白水解酶的口服液或消毒洗液制品中的应用，所述消毒洗液制品选自洗手液、消毒液、消毒凝胶、洗衣液、沐浴液、洗发剂、洗洁精、清洁剂、漱口液或消毒湿纸巾。
8.草棉黄素或没食子酸或槲皮素或芹菜素在制备可抑制新型冠状病毒3CL蛋白水解酶的药物中的应用。
9.草棉黄素或没食子酸或槲皮素或芹菜素在制备可抑制新型冠状病毒3CL蛋白水解酶的口服液或消毒洗液制品中的应用，所述消毒洗液制品选自洗手液、消毒液、消毒凝胶、洗衣液、沐浴液、洗发剂、洗洁精、清洁剂、漱口液或消毒湿纸巾。
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