CN114129616A - 含有蛹虫草提取物和迷迭香提取物的雾化液及其用途 - Google Patents
含有蛹虫草提取物和迷迭香提取物的雾化液及其用途 Download PDFInfo
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- cordyceps militaris
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- A61P37/02—Immunomodulators
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P37/00—Drugs for immunological or allergic disorders
- A61P37/08—Antiallergic agents
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P9/00—Drugs for disorders of the cardiovascular system
- A61P9/10—Drugs for disorders of the cardiovascular system for treating ischaemic or atherosclerotic diseases, e.g. antianginal drugs, coronary vasodilators, drugs for myocardial infarction, retinopathy, cerebrovascula insufficiency, renal arteriosclerosis
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P9/00—Drugs for disorders of the cardiovascular system
- A61P9/14—Vasoprotectives; Antihaemorrhoidals; Drugs for varicose therapy; Capillary stabilisers
Abstract
本发明提供一种雾化液,其特征在于其中含有蛹虫草提取物(高纯度虫草素)和迷迭香提取物。本发明的雾化液可用于预防或治疗肺部疾病、预防或治疗IL‑6和/或IL‑8表达升高导致的疾病、以及保护肺上皮细胞(如BEAS‑2B细胞),例如逆转尼古丁对肺部造成的损伤。
Description
发明领域
本发明属于医药领域,具体涉及一种雾化液,其特征在于其中含有蛹虫草提取物(高纯度虫草素)和迷迭香提取物。本发明的雾化液可用于预防或治疗肺部疾病、预防或治疗IL-6和/或IL-8表达升高导致的疾病、以及保护肺上皮细胞(如BEAS-2B细胞),例如逆转尼古丁对肺部造成的损伤。
背景技术
蛹虫草Cordyceps militaris(L.)Fr.是虫草属Cordyceps的模式种,主要寄生阔叶林或混交林地表土层中鳞翅目昆虫的蛹或幼虫。作为一种重要的经济真菌,蛹虫草富含多种生物活性物质如虫草素、虫草酸、虫草多糖等,表现出广泛的生理活性如抗菌、抗病毒、抗肿瘤、抗氧化、抗炎、免疫调节等,具有较好的医疗保健功效和重要的经济价值。蛹虫草是目前虫草类真菌中研究和应用最为广泛的物种之一,并于2009年被我国卫生部正式批准为新资源食品(后改称新食品原料)。
虫草素亦称虫草菌素、冬虫夏草素、3’-脱氧腺苷,属于嘌呤类生物碱,是蛹虫草的主要活性成分。虫草素作为一种天然活性物质,对人体内脏、神经系统、呼吸系统、免疫系统等都具有较强的保护作用。例如,虫草素对肺部、肝脏、肾脏和胃部等疾病有一定的治疗作用,可以作为化妆品和保健品原料。
由于虫草素脂溶性较差,对其进行体内研究有一定的困难。然而,开发用于临床前和临床研究的虫草素制剂具有很大的发展前景。
迷迭香提取物的主要成分为迷迭香酸(rosmarinic acid,RA)是一种多酚酸,广泛分布于唇形科、紫草科、葫芦科等植物中,是多种药材和中成药的主要成分,具有抗氧化、抗炎、抗肿瘤等多方面药理学活性。
本领域对于能够充分发挥虫草素和迷迭香功能的新制剂存在着迫切的需求。
发明内容
本发明人经过研究,开发了一种含有蛹虫草提取物(高纯度虫草素)和迷迭香提取物的雾化液,并证实其对逆转尼古丁对肺部造成的损伤、抑制IL-6和/或IL-8分泌和保护肺上皮细胞方面存在有益效果。
具体地,在一个方面,本发明提供了一种雾化液,其特征在于其中含有蛹虫草提取物(高纯度虫草素)和迷迭香提取物。
在另一个方面,本发明还提供了蛹虫草提取物(高纯度虫草素)和迷迭香提取物在制备用于预防或治疗肺部疾病的可吸入制剂中的用途。
在另一个方面,本发明还提供了虫草素蛹虫草提取物(高纯度虫草素)和迷迭香提取物在制备用于预防或治疗IL-6和/或IL-8表达升高导致的疾病的可吸入制剂中的用途。
在另一个方面,本发明还提供了虫草素蛹虫草提取物(高纯度虫草素)和迷迭香提取物在制备用于保护肺上皮细胞(如BEAS-2B细胞)的可吸入制剂中的用途。
附图说明
图1:尼古丁盐对BEAS-2B细胞增殖的影响。
图2:蛹虫草提取物对BEAS-2B细胞增殖的影响。
图3:蛹虫草提取物和尼古丁盐同时使用对BEAS-2B细胞增殖的影响。
图4:蛹虫草提取物和尼古丁盐同时使用对BEAS-2B细胞形态的影响(100×)。
图5:蛹虫草提取物和尼古丁盐对BEAS-2B细胞分泌IL-6的影响。
图6:蛹虫草提取物和尼古丁盐对BEAS-2B细胞分泌IL-8的影响。
具体实施方案
本发明具体涉及以下技术方案。
技术方案1.一种雾化液,其特征在于其中含有蛹虫草提取物和迷迭香提取物。
技术方案2.技术方案1的雾化液,其中所述蛹虫草提取物为高纯度虫草素,优选虫草素纯度≥95%。
技术方案3.技术方案1或2的雾化液,其中所述迷迭香提取物为水溶性提取物,优选迷迭香酸纯度≥60%。
技术方案4.技术方案1-3中任一项的雾化液,其中所述蛹虫草提取物的浓度为0.05~0.5g/100g,优选0.1~0.2g/100g,例如0.05g/100g、0.1g/100g、0.15g/100g、0.2g/100g。
技术方案5.技术方案1-4中任一项的雾化液,其中所述迷迭香提取物的浓度为0.1~2g/100g,优选0.1~2g/100g,例如0.1g/100g、0.15g/100g、0.5g/100g、0.75g/100g、0.1g/100g。
技术方案6.技术方案1-5中任一项的雾化液,其中雾化溶剂为醇类溶剂,优选丙二醇、丙三醇或二者的混合物,优选蛹虫草提取物与雾化溶剂的重量比为1:800至1:100,尤其优选1:700至1:400,例如1:700、1:600、1:500、1:400。
技术方案7.技术方案1-6中任一项的雾化液,其中雾化溶剂为醇类溶剂,优选丙二醇、丙三醇或二者的混合物,优选迷迭香与雾化溶剂的重量比为1:600至1:50,尤其优选1:500至1:60,例如1:500、1:400、1:300、1:200、1:150、1:60。
技术方案8.技术方案6或7的雾化液,其中所述雾化溶剂为丙二醇和丙三醇的混合物,二者的重量比为1:10至1:1,优选1:1、1:2、1:3、1:4、1:5。
技术方案9.技术方案1-8中任一项的雾化液,其中还含有香精香料,例如甜味剂、凉味剂、市售食用香精香料、增稠剂。
技术方案10.技术方案9的雾化液,其中所述甜味剂为三氯蔗糖、纽甜、赤藓糖醇、安赛蜜、甜蜜素、糖精钠、阿斯巴甜、阿力甜、新橙皮甙二氢查尔酮、甜菊糖苷、甘草甜素中的一种或多种。
技术方案11.技术方案9的雾化液,其中所述凉味剂为N-乙基-L-薄荷基甲酰胺(WS-23)、N-乙基-5-甲基-2-(1-甲基乙基)环己酰胺(WS-3)、薄荷醇、N-乙基-对薄荷基-3-羧酰胺、N,2,3-三甲基-2-异丙基丁酰胺、N-(乙氧羰基甲基)-对烷-3-甲酰胺、N-(4-甲氧基苯基)-对薄荷基-3-羧酰胺、薄荷脑、薄荷酮、异薄荷酮、异胡薄荷酮、乙酸薄荷酯、左旋香芹酮、乳酸薄荷酯、乙酸异薄荷酯、异戊酸薄荷酯、N-(4-甲氧基苯基)-5-甲基-2-(1-甲基乙基)环己烷甲酰(WS-12)、N-[(乙氧羰基)甲基]-对薄荷烷-3-甲酰胺(WS-5)、3-L-薄荷氧基-1,2-丙二醇(WS-10)中的一种或多种。
技术方案12.技术方案9的雾化液,其中所述市售食用香精香料为:柠檬、甜橙、青苹果、青桔、葡萄、水蜜桃、荔枝、凤梨、樱桃、樱花、哈密瓜、麦香、百香果、苹果、绿豆、红豆、柚子、山楂、芒果、草莓、树莓、蓝莓、椰子、西瓜、香蕉、巧克力、奶油、玫瑰、桂花、青瓜、黑加仑、迷迭香、竹叶香、咖啡、红酒、洛神花、绿茶和红茶中的一种或多种。
技术方案13.技术方案9的雾化液,其中所述增稠剂为羧甲基纤维素钠、藻酸丙二醇酯、卡拉胶、黄原胶、果胶、瓜尔豆胶、刺槐豆胶、明胶、琼脂、海藻酸钠、酪蛋白酸钠、田菁胶中的一种或多种。
技术方案14.技术方案1的雾化液,其中含有以下成分:1-10份蛹虫草提取物溶液,1-10份迷迭香提取物溶液,0-40份香精香料,30-70份丙三醇,其中所述蛹虫草提取物溶液和迷迭香提取物溶液分别为蛹虫草提取物和迷迭香提取物在醇类溶剂,优选丙二醇中的溶液。
技术方案15.技术方案1的雾化液,其含有2%蛹虫草提取物的丙二醇溶液、5%迷迭香提取物的丙二醇溶液、香精香料、丙三醇,其中蛹虫草提取物溶液、迷迭香提取物溶液、香精香料、丙三醇的重量比为5:10:30:55。
技术方案16.技术方案1的雾化液,其含有5%蛹虫草提取物的丙二醇溶液、2%迷迭香提取物的丙二醇溶液、香精香料、丙三醇,其中蛹虫草提取物溶液、迷迭香提取物溶液、香精香料、丙三醇的重量比为4:8:20:68。
技术方案17.技术方案1的雾化液,其含有2%蛹虫草提取物的丙二醇溶液、10%迷迭香提取物的丙二醇溶液、香精香料、丙三醇,其中蛹虫草提取物溶液、迷迭香提取物溶液、香精香料、丙三醇的重量比为5:10:40:45。
技术方案18.技术方案1-17中任一项的雾化液,其通过喷雾、加热雾化形成可供吸入的气溶胶或蒸汽等方式,用于雾化吸入疗法。
技术方案19.技术方案18的雾化液,其中所述加热雾化通过加热雾化装置,例如电子雾化器实现。
技术方案20.技术方案1-19中任一项的雾化液,其用于缓解尼古丁对肺部造成的损伤。
技术方案21.技术方案1-20中任一项的雾化液,其可用于雾化吸入。
技术方案22.蛹虫草提取物(高纯度虫草素)和迷迭香提取物在制备用于预防或治疗肺部疾病的可吸入制剂中的用途。
技术方案23.技术方案22的用途,其中所述肺部疾病为炎性肺部疾病,例如慢性阻塞性肺疾病、肺炎、过敏性哮喘、急性肺损伤、肺纤维化、支气管肺发育不良等。
技术方案24.技术方案22或23的用途,其中所述肺部疾病为尼古丁对肺部造成的损伤。
技术方案25.技术方案22-24中任一项的用途,其中蛹虫草提取物以不超过500μM、优选不超过100μM的浓度使用,优选不超过50μM,优选不超过20μM,优选不超过4μM。
技术方案26.蛹虫草提取物(高纯度虫草素)和迷迭香提取物在制备用于预防或治疗IL-6和/或IL-8表达升高导致的疾病的可吸入制剂中的用途。
技术方案27.技术方案26的用途,其中所述IL-6和/或IL-8表达升高导致的疾病例如动脉粥样硬化、慢性阻塞性肺疾病、自身免疫性疾病(如类风湿关节炎)、炎症性疾病(如血管炎性疾病、胰腺炎、胃肠道炎症、乙型肝炎、前列腺炎)、癌症(如肺癌、黑色素瘤、乳腺癌、胃癌、结肠癌、子宫内膜癌、膀胱癌、胰腺导管腺癌、鼻咽癌)。
技术方案28.技术方案26或27的用途,其中所述疾病由尼古丁诱导。
技术方案29.技术方案26-28中任一项的用途,其中蛹虫草提取物以不超过500μM、优选不超过100μM的浓度使用,优选不超过50μM,优选不超过20μM,优选不超过4μM。
技术方案30.蛹虫草提取物(高纯度虫草素)和迷迭香提取物在制备用于保护肺上皮细胞(如BEAS-2B细胞)的可吸入制剂中的用途。
技术方案31.技术方案30的用途,其中蛹虫草提取物用于防止BEAS-2B细胞炎症或增强BEAS-2B细胞存活。
技术方案32.技术方案30或31的用途,其中所述BEAS-2B细胞炎症或BEAS-2B细胞存活下降与尼古丁摄入有关。
技术方案33.如技术方案30-32中任一项的用途,其中所述BEAS-2B细胞炎症分泌IL-6和/或IL-8炎性驱化性因子。
技术方案34.如技术方案30-33中任一项的用途,其中所述蛹虫草提取物以不超过500μM、优选不超过100μM的浓度使用,优选不超过50μM,优选不超过20μM,优选不超过4μM。
雾化液原料及其配比
本发明提供的雾化液优选以丙二醇和丙三醇作为溶剂,用功能性成分蛹虫草提取物和迷迭香提取物作为溶质,选择性添加香精香料,例如甜味剂、凉味剂、食用香精香料、增稠剂。其优选特征如下:
(1)蛹虫草提取物,优选高纯度蛹虫草提取物,其中虫草素纯度≥95%。
蛹虫草提取物溶液:用丙二醇超声30min溶解。蛹虫草提取物重量占比1~10%。
(2)迷迭香提取物:水溶性提取物,迷迭香酸纯度≥60%。
迷迭香提取物溶液:用丙二醇超声30min溶解。迷迭香提取物重量占比1~10%。
(3)所用丙二醇和丙三醇均为食用级醇类。
(4)香精香料可选择各种香型的市售液体状态食用香精香料。
(5)制备的雾化液组成:1~10份蛹虫草提取物溶液,1~10份迷迭香提取物溶液,0~40份市售香精香料,30~70份丙三醇。
实施例
以下实施例用于例示本发明,并非用于限定本发明的内容。
雾化液的制备方法
实施例1:
(1)准备好2个500ml透明玻璃瓶,配制以下所需溶液:
A.2%蛹虫草提取物:精确称取10g蛹虫草提取物粉末,加490g丙二醇进行溶解,恒温超声振荡器中超声30min混匀;
B.5%迷迭香提取物:精确称取25g迷迭香提取物粉末,加475g丙二醇进行溶解,恒温超声振荡器中超声30min混匀;
(2)按照重量称取所需试剂:2%蛹虫草提取物溶液5g,5%迷迭香提取物溶液10g,市售香精香料30g(柠檬20g,青苹果10g),丙三醇55g。
(3)将调配的组合物放置于恒温超声振荡器中超声15min混合均匀,即得虫草素-迷迭香提取物雾化液。
实施例2:
(1)准备好2个500ml透明玻璃瓶,配制以下所需溶液:
A.5%蛹虫草提取物:精确称取25g蛹虫草提取物粉末,加475g丙二醇进行溶解,恒温超声振荡器中超声30min混匀;
B.2%迷迭香提取物:精确称取10g迷迭香提取物粉末,加490g丙二醇进行溶解,恒温超声振荡器中超声30min混匀;
(2)按照重量称取所需试剂:5%蛹虫草提取物溶液4g,2%迷迭香提取物溶液8g,市售香精香料20g(柠檬3g,青桔5g,百香果2g,迷迭香10g),丙三醇68g。
(3)将调配的组合物放置于恒温超声振荡器中超声15min混合均匀,即得虫草素-迷迭香提取物雾化液。
实施例3:
(1)准备好2个500ml透明玻璃瓶,配制以下所需溶液:
A.2%蛹虫草提取物:精确称取10g蛹虫草提取物粉末,加490g丙二醇进行溶解,恒温超声振荡器中超声30min混匀;
B.10%迷迭香提取物:精确称取50g迷迭香提取物粉末,加450g丙二醇进行溶解,恒温超声振荡器中超声30min混匀;
(2)按照重量称取所需试剂:2%蛹虫草提取物溶液5g,10%迷迭香提取物溶液10g,市售香精香料40g(水蜜桃30g,茉莉2g,绿茶8g),丙三醇45g。
(3)将调配的组合物放置于恒温超声振荡器中超声15min混合均匀,即得虫草素-迷迭香提取物雾化液。
实施例4:蛹虫草和迷迭香提取物雾化液的受试者评价
为评估本发明的蛹虫草和迷迭香提取物雾化液的使用感受,在吸烟受试者中对比电子烟液进行了评价。
(1)受试对象的选择:选取三组烟龄存在差异的受试群体。
A.选取烟龄在5年以内的吸烟受试者;B.选取烟龄在5~10年的吸烟受试者;C.选取烟龄在10~30年的吸烟受试者。受试者可能会有咽喉不适,如咽痒、咽干、咽痛、咳嗽、异物感,鼻塞,口腔异味等情况(至少一项),但均无严重心、肝、肾等主要脏器疾患等。受试对象每组18例,男女比例为1:1。
(2)实验方法:将A、B、C三组受试对象再各分为3组,每组6例,每组分别吸入实施例1~3的雾化液(配有统一的电子烟装置),每日抽吸30口,早中晚各10口,持续3天。
(3)受试者感官评价内容
表1受试者评价内容
(4)抽吸3日后的结果见表2~4。表2~4显示了实施例1~3的雾化液的感官评价及对咽喉不适、口腔异味和鼻塞的改善效果。
表2不同实施例的雾化液对于A组受试者的效果
表3不同实施例的雾化液对于B组受试者的效果
表4不同实施例的雾化液对于C组受试者的效果
由表2可知,对于抽吸年限在5年以下的人群,实施例2的雾化液总体评价较好;实施例1和实施例2的雾化液对咽喉不适和鼻塞的改善优于实施例3;实施例1和实施例3雾化液对于口腔异味的改善优于实施例2。由表3可知,对于抽吸年限在5~10年的人群,所有雾化液总体评价相差不大;实施例1和实施例2的雾化液对咽喉不适和鼻塞的改善优于实施例3;由表4可知,对于抽吸年限在10~30年的人群,所有雾化液总体评价相差不大;实施例1和实施例2的雾化液对咽喉不适和鼻塞的改善优于实施例3;实施例1和实施例3雾化液对于口腔异味的改善优于实施例2。
对咽喉不适和鼻塞的改善推测原因是蛹虫草提取物的功效。对于口腔异味的改善推测可能是由于迷迭香本身的气味的因素,以及蛹虫草提取物的杀菌和免疫调节作用。由此可见,蛹虫草提取物和迷迭香提取物对于抽吸电子烟造成的口腔异味、鼻塞和咽喉不适具有保健作用。
实施例5:蛹虫草提取物的细胞学研究
5.1蛹虫草提取物对BEAS-2B细胞(人正常肺上皮细胞)存活率的影响设计本实施例用于评估蛹虫草提取物能否逆转尼古丁的不良作用。
实验过程
1640培养基中添加10%胎牛血清、1%青霉素和链霉素构成完全培养基。BEAS-2B细胞用完全培养基放置于37℃、5%CO2饱和湿度培养箱内培养,根据细胞生长状态,按(1:2):(1-4)传代。取对数生长期的细胞,0.25%-EDTA胰酶消化,800rpm 4℃离心5min,培养基重悬细胞后,显微镜下细胞计数后,细胞浓度调整为1×105个/mL,接种至96孔细胞培养板,每孔加1×104个细胞。细胞贴壁生长过夜后,分别加入含药的完全培养基,每组3个复孔。培养24h后,每孔加入10μL CCK-8溶液。在细胞培养箱内继续孵育3小时。酶标仪A450nm测定吸光度。根据公式细胞存活率(%)=(A药物-A空白)/(A对照-A空白)*100%计算细胞存活率。
(1)不同浓度尼古丁盐和蛹虫草提取物分别对细胞增殖的影响:尼古丁盐(纯度:99%)0mM、3.125mM、6.25mM、12.5mM、25mM、50mM和100mM;蛹虫草提取物(虫草素纯度:98%)浓度设置为0μM、0.8μM、4μM、20μM、100μM、500μM和2500μM,同时设有空白组、未加药的对照组。
(2)蛹虫草提取物和尼古丁同时使用时对细胞增殖的影响:蛹虫草提取物(4μM、20μM)组,尼古丁盐(25mM)组及蛹虫草提取物和尼古丁盐同时使用组(蛹虫草提取物4μM+尼古丁盐25mM,蛹虫草提取物20μM+尼古丁盐25mM),同时设有空白组、未加药的对照组。
实验结果和分析
尼古丁盐对BEAS-2B细胞增殖的影响
由图1可知,尼古丁盐浓度在0~100mM范围内,对BEAS-2B细胞增殖均有抑制作用。尼古丁盐浓度在25~100mM时,BEAS-2B细胞增殖率小于50%(即仅为正常增殖的不到一半)。
由图2可知,蛹虫草提取物浓度在0~2500μM(0~2.5mM)范围内,对BEAS-2B细胞增殖可能有抑制作用,尤其是浓度大于500μM时。但是,蛹虫草提取物浓度在0~500μM时,BEAS-2B细胞增殖率大于80%,此时蛹虫草提取物对细胞增殖率影响较小。
由图3可知,蛹虫草提取物浓度在4μM和20μM时,BEAS-2B细胞增殖率均大于96%,参见(2)和(3)。尼古丁盐浓度在25mM时,BEAS-2B细胞增殖率小于50%,参见(1)。然而,在25mM浓度的尼古丁盐基础上再增加4μM或20μM的蛹虫草提取物,则可以逆转尼古丁盐对BEAS-2B细胞增殖率的不利影响,参见(4)和(5)。具体而言,与25mM尼古丁盐作用相比,尼古丁盐25mM+蛹虫草提取物4μM时,BEAS-2B细胞增殖率增加约20%,对比(1)和(4);尼古丁盐25mM+蛹虫草提取物20μM时,BEAS-2B细胞增殖率增加约30%,对比(1)和(5)。可见,蛹虫草提取物可以逆转尼古丁对于肺部正常细胞(BEAS-2B细胞)的损伤,对于肺部正常细胞具有保护作用。
由图4可知,蛹虫草提取物浓度在4μM和20μM时,BEAS-2B细胞形态基本上没有变化。尼古丁盐浓度在25mM时,50%BEAS-2B细胞皱缩,趋近死亡。与尼古丁盐25mM单独作用相比,尼古丁盐25mM+蛹虫草提取物4μM和尼古丁盐25mM+蛹虫草提取物20μM时,BEAS-2B细胞数量皱缩和死亡变少。再次证明蛹虫草提取物对于肺部正常细胞(BEAS-2B细胞)具有保护作用。
5.2蛹虫草提取物和尼古丁盐对BEAS-2B细胞(人正常肺上皮细胞)炎性因子的影响
设计本实施例用于评估蛹虫草提取物的抗炎作用。
实验方法与过程
1640培养基中添加10%胎牛血清、1%青霉素和链霉素构成完全培养基。BEAS-2B细胞用完全培养基放置于37℃、5%CO2饱和湿度培养箱内培养,根据细胞生长状态,按(1:2):(1-4)传代。取对数生长期的细胞,0.25%-EDTA胰酶消化,800rpm 4℃离心5min,培养基重悬细胞后,显微镜下细胞计数后,细胞浓度调整为1×105个/mL,接种至6孔细胞培养板,每孔加1×104个细胞。细胞贴壁生长过夜后,分别加入含药的完全培养基,每组3个复孔。组别:尼古丁盐(纯度:99%)浓度设置为12.5mM、25mM、50mM和100mM;蛹虫草提取物(虫草素纯度:98%)浓度设置为6.25μM、12.5μM,同时设有空白组、正常细胞对照组。培养24h后,收取培养基,离心后取上清。参照试剂盒说明书检测炎性因子IL-6、IL-8水平(碧云天PI330Human IL-6ELISA Kit、PI640 Human IL-8ELISA Kit)。
实验结果与分析
蛹虫草提取物和尼古丁盐对BEAS-2B细胞分泌IL-6的影响
白细胞介素6(interleukin 6,IL-6)是一种典型的细胞因子,在炎症反应、免疫应答和造血方面具有多效性。IL-6主要由巨噬细胞产生,通过去除传染性病原体、诱导受损组织愈合、急性期免疫反应以及凝血级联反应从而执行保护功能。由图5可知,与正常细胞组相比,12.5mM、25mM、50mM和100mM尼古丁盐组BEAS-2B细胞分泌IL-6明显增多,6.25μM和12.5μM蛹虫草提取物组IL-6水平却有降低趋势。可见,蛹虫草提取物可以抑制IL-6的释放。
蛹虫草提取物和尼古丁盐对BEAS-2B细胞分泌IL-8的影响
白细胞介素-8(interleukin-8,IL-8),又称趋化因子CXCL8(CXCL8),是一种炎性趋化性因子。IL-8通过受体CXCR1和CXCR2产生多种生物学功能,包括促炎性细胞趋化、促血管生成、促有丝分裂和诱导细胞增殖等。由图6可知,与正常细胞组相比,12.5mM、25mM、50mM尼古丁盐组BEAS-2B细胞分泌IL-8明显增多,100mM尼古丁盐组BEAS-2B细胞分泌IL-8没有明显变化,可能与其他细胞通路参与调节有关。6.25μM和12.5μM蛹虫草提取物组IL-8水平却有降低趋势。可见,蛹虫草提取物可以抑制IL-8的释放。
综上可知,尼古丁盐在一定范围内可能会促进炎性因子IL-6和IL-8的释放,而一定范围的蛹虫草提取物可以抑制其释放。
以上内容是结合具体的优选实施方式对本发明所作的进一步详细说明,不能认定本发明的具体实施只局限于这些说明。对于本发明所属技术领域的普通技术人员来说,在不脱离本发明构思的前提下,还可以做出若干简单推演或替换,都应当视为属于本发明的保护范围。
Claims (34)
1.一种雾化液,其特征在于其中含有蛹虫草提取物和迷迭香提取物。
2.权利要求1的雾化液,其中所述蛹虫草提取物为高纯度虫草素,优选虫草素纯度≥95%。
3.权利要求1或2的雾化液,其中所述迷迭香提取物为水溶性提取物,优选迷迭香酸纯度≥60%。
4.权利要求1-3中任一项的雾化液,其中所述蛹虫草提取物的浓度为0.05~0.5g/100g,优选0.1~0.2g/100g,例如0.05g/100g、0.1g/100g、0.15g/100g、0.2g/100g。
5.权利要求1-4中任一项的雾化液,其中所述迷迭香提取物的浓度为0.1~2g/100g,优选0.1~2g/100g,例如0.1g/100g、0.15g/100g、0.5g/100g、0.75g/100g、0.1g/100g。
6.权利要求1-5中任一项的雾化液,其中雾化溶剂为醇类溶剂,优选丙二醇、丙三醇或二者的混合物,优选蛹虫草提取物与雾化溶剂的重量比为1:800至1:100,尤其优选1:700至1:400,例如1:700、1:600、1:500、1:400。
7.权利要求1-6中任一项的雾化液,其中雾化溶剂为醇类溶剂,优选丙二醇、丙三醇或二者的混合物,优选迷迭香与雾化溶剂的重量比为1:600至1:50,尤其优选1:500至1:60,例如1:500、1:400、1:300、1:200、1:150、1:60。
8.权利要求6或7的雾化液,其中所述雾化溶剂为丙二醇和丙三醇的混合物,二者的重量比为1:10至1:1,优选1:1、1:2、1:3、1:4、1:5。
9.权利要求1-8中任一项的雾化液,其中还含有香精香料,例如甜味剂、凉味剂、市售食用香精香料、增稠剂。
10.权利要求9的雾化液,其中所述甜味剂为三氯蔗糖、纽甜、赤藓糖醇、安赛蜜、甜蜜素、糖精钠、阿斯巴甜、阿力甜、新橙皮甙二氢查尔酮、甜菊糖苷、甘草甜素中的一种或多种。
11.权利要求9的雾化液,其中所述凉味剂为N-乙基-L-薄荷基甲酰胺(WS-23)、N-乙基-5-甲基-2-(1-甲基乙基)环己酰胺(WS-3)、薄荷醇、N-乙基-对薄荷基-3-羧酰胺、N,2,3-三甲基-2-异丙基丁酰胺、N-(乙氧羰基甲基)-对烷-3-甲酰胺、N-(4-甲氧基苯基)-对薄荷基-3-羧酰胺、薄荷脑、薄荷酮、异薄荷酮、异胡薄荷酮、乙酸薄荷酯、左旋香芹酮、乳酸薄荷酯、乙酸异薄荷酯、异戊酸薄荷酯、N-(4-甲氧基苯基)-5-甲基-2-(1-甲基乙基)环己烷甲酰(WS-12)、N-[(乙氧羰基)甲基]-对薄荷烷-3-甲酰胺(WS-5)、3-L-薄荷氧基-1,2-丙二醇(WS-10)中的一种或多种。
12.权利要求9的雾化液,其中所述市售食用香精香料为:柠檬、甜橙、青苹果、青桔、葡萄、水蜜桃、荔枝、凤梨、樱桃、樱花、哈密瓜、麦香、百香果、苹果、绿豆、红豆、柚子、山楂、芒果、草莓、树莓、蓝莓、椰子、西瓜、香蕉、巧克力、奶油、玫瑰、桂花、青瓜、黑加仑、迷迭香、竹叶香、咖啡、红酒、洛神花、绿茶和红茶中的一种或多种。
13.权利要求9的雾化液,其中所述增稠剂为羧甲基纤维素钠、藻酸丙二醇酯、卡拉胶、黄原胶、果胶、瓜尔豆胶、刺槐豆胶、明胶、琼脂、海藻酸钠、酪蛋白酸钠、田菁胶中的一种或多种。
14.权利要求1的雾化液,其中含有以下成分:1-10份蛹虫草提取物溶液,1-10份迷迭香提取物溶液,0-40份香精香料,30-70份丙三醇,其中所述蛹虫草提取物溶液和迷迭香提取物溶液分别为蛹虫草提取物和迷迭香提取物在醇类溶剂,优选丙二醇中的溶液。
15.权利要求1的雾化液,其含有2%蛹虫草提取物的丙二醇溶液、5%迷迭香提取物的丙二醇溶液、香精香料、丙三醇,其中蛹虫草提取物溶液、迷迭香提取物溶液、香精香料、丙三醇的重量比为5:10:30:55。
16.权利要求1的雾化液,其含有5%蛹虫草提取物的丙二醇溶液、2%迷迭香提取物的丙二醇溶液、香精香料、丙三醇,其中蛹虫草提取物溶液、迷迭香提取物溶液、香精香料、丙三醇的重量比为4:8:20:68。
17.权利要求1的雾化液,其含有2%蛹虫草提取物的丙二醇溶液、10%迷迭香提取物的丙二醇溶液、香精香料、丙三醇,其中蛹虫草提取物溶液、迷迭香提取物溶液、香精香料、丙三醇的重量比为5:10:40:45。
18.权利要求1-17中任一项的雾化液,其通过喷雾、加热雾化形成可供吸入的气溶胶或蒸汽等方式,用于雾化吸入疗法。
19.权利要求18的雾化液,其中所述加热雾化通过加热雾化装置,例如电子雾化器实现。
20.权利要求1-19中任一项的雾化液,其用于缓解尼古丁对肺部造成的损伤。
21.权利要求1-20中任一项的雾化液,其可用于雾化吸入。
22.蛹虫草提取物(高纯度虫草素)和迷迭香提取物在制备用于预防或治疗肺部疾病的可吸入制剂中的用途。
23.权利要求22的用途,其中所述肺部疾病为炎性肺部疾病,例如慢性阻塞性肺疾病、肺炎、过敏性哮喘、急性肺损伤、肺纤维化、支气管肺发育不良等。
24.权利要求22或23的用途,其中所述肺部疾病为尼古丁对肺部造成的损伤。
25.权利要求22-24中任一项的用途,其中蛹虫草提取物以不超过500μM、优选不超过100μM的浓度使用,优选不超过50μM,优选不超过20μM,优选不超过4μM。
26.蛹虫草提取物(高纯度虫草素)和迷迭香提取物在制备用于预防或治疗IL-6和/或IL-8表达升高导致的疾病的可吸入制剂中的用途。
27.权利要求26的用途,其中所述IL-6和/或IL-8表达升高导致的疾病例如动脉粥样硬化、慢性阻塞性肺疾病、自身免疫性疾病(如类风湿关节炎)、炎症性疾病(如血管炎性疾病、胰腺炎、胃肠道炎症、乙型肝炎、前列腺炎)、癌症(如肺癌、黑色素瘤、乳腺癌、胃癌、结肠癌、子宫内膜癌、膀胱癌、胰腺导管腺癌、鼻咽癌)。
28.权利要求26或27的用途,其中所述疾病由尼古丁诱导。
29.权利要求26-28中任一项的用途,其中蛹虫草提取物以不超过500μM、优选不超过100μM的浓度使用,优选不超过50μM,优选不超过20μM,优选不超过4μM。
30.蛹虫草提取物(高纯度虫草素)和迷迭香提取物在制备用于保护肺上皮细胞(如BEAS-2B细胞)的可吸入制剂中的用途。
31.权利要求30的用途,其中蛹虫草提取物用于防止BEAS-2B细胞炎症或增强BEAS-2B细胞存活。
32.权利要求30或31的用途,其中所述BEAS-2B细胞炎症或BEAS-2B细胞存活下降与尼古丁摄入有关。
33.如权利要求30-32中任一项的用途,其中所述BEAS-2B细胞炎症分泌IL-6和/或IL-8炎性驱化性因子。
34.如权利要求30-33中任一项的用途,其中所述蛹虫草提取物以不超过500μM、优选不超过100μM的浓度使用,优选不超过50μM,优选不超过20μM,优选不超过4μM。
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刘慧: "迷迭香功效成分提取及药理作用", 《农产品加工》 * |
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