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CN112137935B - 一种组合物及其用途 - Google Patents

一种组合物及其用途 Download PDF

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CN112137935B
CN112137935B CN202011159114.2A CN202011159114A CN112137935B CN 112137935 B CN112137935 B CN 112137935B CN 202011159114 A CN202011159114 A CN 202011159114A CN 112137935 B CN112137935 B CN 112137935B
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extract
hemp
sweet wormwood
composition
cannabis
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CN112137935A (zh
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王玉亮
唐克轩
孙小芬
刘品
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Suzhou Tangji Biological Technology Co ltd
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Suzhou Tangji Biological Technology Co ltd
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Abstract

本申请公开了一种组合物，其特征在于，所述组合物包括大麻提取物和青蒿提取物，所述大麻提取物和青蒿提取物的比例为10：1至1：10。本申请还公开了所述组合物在在制备药品中的用途，所述药品用于痤疮、炎症反应和/或过敏性红斑、抗菌。

Description

一种组合物及其用途
技术领域
本发明涉及生物医药领域，尤其涉及一种大麻青蒿组合物及其用途。
背景技术
皮肤是人体面积最大的组织，位于体表最外层，具有屏障、吸收、分泌、排泄、代谢、免疫、体温调节及感觉功能。其中，皮肤屏障是基础功能，包括表皮渗透屏障、免疫屏障、色素屏障等，同时，皮肤微生态及酸性环境与皮肤屏障之间也有着千丝万缕的联系，共同维持皮肤正常生理代谢。
表皮渗透屏障功能，对外可以抵御抗原物质、微生物、日光等的侵袭，对内可防止体内营养物质、水分的丢失。表皮渗透屏障受损，经表皮水分流失(transepidermalwaterloss，TEWL)增加，皮肤则出现干燥、脱屑。同时，对外界抗原及微生物的抵御能力下降，可导致多种皮肤病的发生。表皮渗透屏障与免疫屏障、酸性屏障及微生态共同构成一个整体防御系统，表皮微生态、pH值可影响表皮渗透屏障，表皮渗透屏障受损，又可影响其他屏障功能。
表皮渗透屏障受多种因素影响，如年龄等内在因素，以及紫外线等外在因素。表皮渗透屏障受损，老年人容易出现瘙痒、皮肤炎症反应，婴幼儿易患有特应性皮炎。紫外线是影响表皮渗透屏障的主要外在因素。当前诸多研究显示特应性皮炎、多形性日光疹、痤疮、玫瑰痤疮、湿疹、光老化等皮肤病的发生都与表皮渗透屏障受损有关，提示临床治疗这类皮肤病时需注意修复受损的表皮渗透屏障功能。
发明内容
为实现上述目的，本申请提供了一种组合物，其特征在于，其包括大麻提取物和青蒿提取物，所述大麻提取物和青蒿提取物的比例为10：1至1：10。
在某些实施方式中，所述组合物具有至少如下功能之一：
a)抗氧化作用；
b)防御UVA和/或UVB对皮肤的损害；
c)缓解炎症反应或过敏反应；
d)抑制细菌生长。
在某些实施方式中，所述大麻提取物来自大麻花叶、大麻果仁、大麻籽油和/或大麻槿茎粉。
在某些实施方式中，所述青蒿提取物来自青蒿叶。
在某些实施方式中，所述大麻提取物为大麻二酚。
在某些实施方式中，所述大麻二酚的纯度不低于95％(质量比)。
在某些实施方式中，所述青蒿提取物为青蒿素。
在某些实施方式中，所述青蒿素的纯度不低于95％(质量比)。
在某些实施方式中，所述大麻提取物的制备方法为：选取9-10月大麻花叶，制成干制品，取所述干制品加入10倍重量的95％的乙醇，超声提取，分离提取液，真空浓缩干燥。
在某些实施方式中，所述青蒿提取物的制备方法为：选取9-10月青蒿叶，制成干制品，取所述干制品加入10倍重量的95％的乙醇，超声提取分离提取液，真空浓缩干燥。
在某些实施方式中，所述大麻提取物和青蒿提取物的比例为10：1。
在某些实施方式中，所述大麻提取物和青蒿提取物的比例为5：1。
在某些实施方式中，所述大麻提取物和青蒿提取物的比例为1：1。
在某些实施方式中，所述大麻提取物和青蒿提取物的比例为1：5。
在某些实施方式中，所述大麻提取物和青蒿提取物的比例为1：10。
在某些实施方式中，所述组合物的制备方法为：a)将大麻果实和青蒿叶片1：1混合，蒸馏法制备挥发油；或者，b)大麻叶片和青蒿叶片2：1混合，40℃-50℃干燥，超微粉碎后过300目筛。
另一方面，本申请还提供本申请所述的组合物在制备药品中的用途，所述药品用于预防、缓解或治疗痤疮、炎症反应和/或过敏性红斑、抗菌。
在某些实施方式中，所述药品被配置为经皮肤施用。
在某些实施方式中，所述药品包括合适的辅料。
在某些实施方式中，所述药品包括合适的辅料。
在某些实施方式中，所述药品中所述组合物的质量分数为0.5％至10％。
在某些实施方式中，所述药品被配置为霜剂、乳剂、悬浮剂、水剂、粉剂、面膜。
本申请提供的大麻提取物和青蒿提取物的组合，具有良好的抗氧化、缓解炎症反应或过敏反应以及抑制细菌生长的功效，并且其效果显著优于大麻提取物或青蒿提取物的单用效果，具有协同作用，取得了预料不到的技术效果。
具体实施方式
以下将结合实施例对本发明作进一步地说明，应理解这些实施例仅作为例证的目的，本发明可以通过许多不同形式的实施例来得以体现，本发明的保护范围并非仅限于文中提到的实施例。
实施例1制备大麻和青蒿的提取物
大麻提取物：选取9-10月间大麻现蕾时花叶，自然晒干，或者鼓风烘箱50℃烘干至含水量低于10％。使用具塞三角瓶称取干叶100克，加入1升95％的乙醇，超声提取(上海之信超声发生器)，设定提取功率为60K,温度60℃，提取30min，过滤分离提取液，真空浓缩干燥至无液体流出。
大麻二酚制备方法：将上述大麻提取物，用500ml氯仿溶解，用纯水萃取4次(每次75ml)。氯仿溶解部分(3g)用硅胶层析板层析,以含4％乙醇的氯仿展开,用大麻二酚标准品确定展开位置，刮下相应位置硅胶，氯仿溶解，加入20％的乙醇，结晶得到大麻二酚含量为95％以上。
青蒿提取物：选取9-10月间现蕾前的青蒿叶片，自然晒干，或者鼓风烘箱50℃烘干至含水量低于10％。使用具塞三角瓶称取干叶100克，加入1升95％的乙醇，超声提取(上海之信超声发生器)，设定提取功率为60K,温度60度，提取30min，过滤分离提取液，真空浓缩干燥至无液体流出。
青蒿素制备方法：将上述青蒿提取物用石油醚溶解后，过硅胶柱吸附，用2倍柱体积的石油醚冲洗后，用石油醚：丙酮3：1的洗脱液洗脱，收集该部分洗脱液，真空干燥后，用乙醇溶解，结晶，可得到青蒿素含量为95％以上的青蒿素提取物。
实施例2抗氧化活性检测
目前已建立的化学模型法有Trolox当量的抗氧化能力(trolox equivalentantioxidant capacity，TEAC)、1,1-二苯基-2-三硝基苯肼(DPPH)、总自由基捕获抗氧化参数(total radical-trapping antioxidant parameter，TRAP)和总氧自由基清除能力(total oxyradical scavenging capacity，TOSC)等方法。这些方法仅针对单一植物天然成分的理化特性(如水溶性或脂溶性)进行抗氧化能力测定，而且不采用容易被自由基氧化的脂类作为标准参照物，很难真实反映出植物天然活性成分的抗氧化活性。氧自由基吸收能力法(Oxygen radical absorbance capacity,ORAC)可以确定抗氧化物对偶氮二脒基丙烷盐酸盐(ABAP)产生的过氧化自由基的抑制清除效果，是一个经典的通过氢原子转移来清除由ABAP产生的ROO-、OH-等自由基的能力来评价抗氧化剂的抗氧化活性的方法，使用荧光指标剂的衰减曲线下面积(AUC)做为定量的手段。与其他抗氧化评价方法相比，该方法能够将抗氧化剂对自由基的抑制时间和抑制作用结合成一个单独的量。ORAC法是目前国际上常用的一种评价抗氧化剂活性的方法，现已成为美国农业部、美国国立卫生院、美国食品药品监督管理局(FDA)评价食品抗氧化能力的重要标准。美国、欧洲、日本等国家的食品、功能食品行业普遍采用ORAC作为功能食品的重要评价标准。
本实施例采用氧自由基清除测定法(ORAC)，方法参照文献(余欣珂,明建,支玲,李苇舟,闫慧明,王启明,赵吉春.真空冷冻干燥对菊花多酚含量及其抗氧化活性的影响[J].食品与机械,2020,36(6):138-144.)对实施例1中所得的大麻提取物和青蒿提取物单独使用及其配方使用的抗氧化活性进行比较，结果见表1。
表1抗氧化活性
Figure GDA0003774431960000041
如表1中所示，复方的抗氧化活性优于青蒿或大麻单独使用的效果，也优于2种的简单加和，具有协同增效作用。
实施例3大麻及青蒿提取物活性成分抗炎抗过敏作用检测
使用实施例1中制备的大麻二酚和青蒿素验证其抗炎抗过敏作用。肥大细胞是由造血祖细胞衍化而来，广泛分布于结缔组织，皮肤上皮下方尤其多，其细胞较大，细胞质内含有嗜染性的粗大颗粒，当受到刺激，细胞内储存的媒介物质(组胺、肝素、Protease、加水分解酵素等)将被释放出来，参与多种免疫反应，包括炎症反应和过敏反应。炎症反应与肥大细胞脱颗粒密切相关，也是过敏反应过程中的关键步骤，抑制肥大细胞脱颗粒和释放组胺可减轻炎症反应的强度，因此可用以验证某种活性成分的抗炎和抗过敏的作用。结果见表2。
表2抗炎抗过敏结果
Figure GDA0003774431960000042
表2中，肥大细胞脱颗粒率(％)＝(脱颗粒肥大细胞数/肥大细胞总数)×100；组胺释放率(％)＝(组胺释放量/总的组胺释放量)×100；复方1：大麻二酚：青蒿素1：1；复方2：大麻二酚：青蒿素10：1；复方3：大麻二酚：青蒿素1：10；DMSO:二甲基亚砜；LPS：脂多糖。
结果表明青蒿素和大麻二酚均有较好的抗炎抗过敏效果，且两者以不同比例联用有协同增效的作用。
实施例4大麻及青蒿提取物活性成分抑菌作用检测
将灭菌固体LB培养基加热至熔化，待冷却至50℃时，每20ml培养基倒入无菌培养皿中。待平板自然晾干后，分别移取0.1ml待测药品(含20％的实施例1制备的大麻或青蒿提取物)，0.1mL供试菌悬液(106个/ml)，均匀涂布于加药平板上，每个处理3次重复，涂布30min后倒置。另设置对照组，涂菌，以等量无菌水(溶解药品物质)代替药液。上述操作在超净工作台内进行。将做好的细菌平板在37℃恒温培养24h，统计菌落个数，按照下式计算抑菌率。
Figure GDA0003774431960000051
表3
Figure GDA0003774431960000052
结果表明，大麻青蒿提取物的组合物具有更好的抑菌效果。
实施例5组合物的制备
制备青蒿大麻组合物，大麻果实和青蒿叶片1：1混合，采用蒸馏法制备挥发油。
实施例6组合物的制备
制备青蒿大麻组合为，大麻叶片和青蒿叶片2：1混合，45度干燥2小时，超微粉碎后过300目筛。
以上详细描述了本发明的较佳具体实施例。应当理解，本领域的普通技术无需创造性劳动就可以根据本发明的构思作出诸多修改和变化。因此，凡本技术领域中技术人员依本发明的构思在现有技术的基础上通过逻辑分析、推理或者有限的实验可以得到的技术方案，皆应在由权利要求书所确定的保护范围内。

Claims (3)

1.组合物在制备药品中的用途，所述药品用于预防、缓解或治疗炎症反应和/或过敏性红斑、抗菌；所述组合物包括大麻提取物和青蒿提取物，所述大麻提取物和青蒿提取物的比例为10：1至1：10；所述大麻提取物的制备方法为：选取9-10月大麻花叶，制成干制品，取所述干制品加入10倍重量的95％的乙醇，超声提取，分离提取液，真空浓缩干燥；所述青蒿提取物的制备方法为：选取9-10月青蒿叶，制成干制品，取所述干制品加入10倍重量的95％的乙醇，超声提取分离提取液，真空浓缩干燥。
2.如权利要求1所述的用途，其特征在于，所述大麻提取物为大麻二酚。
3.如权利要求1所述的用途，其特征在于，所述青蒿提取物为青蒿素。
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