CN110638688A - 一种青蒿素负载香菇菌多糖复合物的制备方法及应用 - Google Patents
一种青蒿素负载香菇菌多糖复合物的制备方法及应用 Download PDFInfo
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Abstract
本发明属于生物化学技术领域，具体涉及一种青蒿素负载香菇菌多糖复合物的制备方法及应用。青蒿素负载香菇菌多糖复合物包括按重量百分含量组分的原料：香菇菌多糖，含量80‑99％；青蒿素，含量1％‑20％。本发明的青蒿素负载香菇菌多糖复合物制备过程，可以使得青蒿素的稳定性得到很大提高，而且在化妆品中实现缓慢释放。
Description
技术领域
本发明属于生物化学技术领域，具体涉及一种青蒿素负载香菇菌多糖复合物的制备方法及应用。
背景技术
香菇菌多糖，它既是水溶性的，又是电中性的，因此更加适合于化妆品的应用，其拥有优异的抗衰老功效，能够抚平细小皱纹，改善皮肤纹理度。赋予了产品良好的透皮吸收性能，可作为活性成份的释放载体，给予皮肤如丝绸般滋润光滑的触感。
青蒿素，又称黄花素,是从植物黄花蒿茎中提取的有过氧基团的倍半萜内脂药物。
痤疮等皮肤问题可由炎症引起，通过抑制炎症并改善皮肤状态可有效改善痤疮、皮肤衰老等问题。在护肤产品中，往往需要搭配多种来产生协同作用起到抑制炎症同时改善皮肤。
发明内容
本公开的一方面解决的一个技术问题是提供一种青蒿素负载香菇菌多糖复合物的制备方法及应用，目的在于融合香菇菌多糖和青蒿素，使二者协同作用用于护肤产品及药物的研发。
为实现上述目的，本发明采用如下技术方案，本发明青蒿素负载香菇菌多糖复合物，包括按重量百分含量组分的原料：香菇菌多糖，含量80-99％；青蒿素，含量1％-20％。
进一步的，所述香菇菌多糖，又称香菇多糖，Lentinan，CAS:37339-90-5，分子量：1152.9995，灰白色粉末。
进一步的，所述青蒿素，又称黄华素，Artemisinine，CAS:63968-64-9，分子量：282.33，无色针状晶体。
一种上述的青蒿素负载香菇菌多糖复合物的制备方法，所述香菇菌多糖采用生物发酵后提取方式制备，然后与青蒿素通过化学合成方式获得青蒿素负载香菇菌多糖复合物复合物，包括以下步骤：
步骤一：以新鲜绿茶茶叶为主，蔗糖、蛋白胨、磷酸氢钾等为辅，按照茶叶与水重量比为1:1的比例使茶叶和水充分混匀；在121℃高温条件下充分灭菌20-30分钟后得到茶叶培养基；将所述茶叶培养基与香菇菌丝体共培养；对香菇菌丝体的发酵产物水提醇沉分离提取，最终精制得到香菇菌多糖；
步骤二：香菇菌多糖与青蒿素以相应的比例混合在去离子水中；加入与青蒿素等当量的二环己基碳二亚胺作为缩合剂；在室温下搅拌反应24h；透析24小时，除去多余二环己基碳二亚胺以及未反应的青蒿素；冷冻干燥，最终得到相应的青蒿素负载香菇菌多糖复合物。
进一步的，所述香菇菌丝体采用的香菇菌品种不限于某单一品种的香菇菌。
所述的青蒿素负载香菇菌多糖复合物主要以水性糊剂或凝胶或液体的形式，例如水性液体的形式。
优选地，将青蒿素负载香菇菌多糖复合物配制为含水洗液、油包水或水包油乳液、油或油.醇洗液、阴离子或非离子两亲脂质的囊状分散体、含水凝胶、水.醇凝胶、醇凝胶或油.醇凝胶、固体棒或气溶胶。
一种青蒿素负载香菇菌多糖复合物用于抗炎、抗氧化衰老、护肤的制剂。
优先地，所述剂型为临床上可接受的任一剂型。
进一步的，所述青蒿素负载香菇菌多糖复合物为所述抗炎、抗氧化衰老或护肤的唯一活性成分，或该复合物与其他物质一起制备所述抗炎、抗氧化衰老或护肤的制剂。
本发明与现有技术相比，本发明的青蒿素负载香菇菌多糖复合物制备过程，可以使得青蒿素的稳定性得到很大提高，而且在化妆品中实现缓慢释放。
附图说明
图1.DT002对炎症因子TNF-α和IL-1β的影响；
图2.DT002对小鼠足趾肿胀的影响。
具体实施方式
下面将结合具体的实施方案对本发明进行进一步的解释，但并不局限本发明，在本领域普通技术人员所具备的知识范围内，还可以在不脱离本发明宗旨的前提下作出各种变化。
下述实施例中，如无特殊说明，所用方法为常规方法，所用试剂都来源于市售商品。
实施例1.青蒿素负载香菇菌多糖复合物对炎症因子TNF-α和IL-1β的影响检测
检测方法：
小鼠巨噬细胞RAW264.7接种于12孔板内，37℃，5％CO2培养箱培养24h后，加入40mg/L脂多糖(LPS)作用4h诱导炎症反应，适当浓度的DT002、GK2、Drag和Hydr于LPS加入前2h加入。LPS作用4h后，收集细胞上清，按照ELISA试剂盒说明书测定TNF-α和IL-1β的含量。
结果如图1所示。
DT002：青蒿素负载香菇菌多糖复合物；LPS：脂多糖；GK2：甘草酸二钾；Drag：红没药醇；Hydr：氢化可的松。(n＝3，**P<0.01)
结论：
DT002显著抑制LPS诱导引起的炎症因子TNF-α和IL-1β含量的上升，且效果显著优于阳性药甘草酸二钾和红没药醇，其抑炎效果与糖皮质激素氢化可的松效果相当。
实施例2.对小鼠足趾肿胀的影响检测
检测方法：
1％羧甲基纤维素钠配药，使成凝胶状，并在每鼠右下足趾正反两面均匀涂抹药物。半小时后，每鼠左右两足趾均皮下注射1％角叉菜胶约0.025mL，空白对照组小鼠不做处理。5h后脱颈处死，沿踝关节减下左右两足，称重，计算肿胀度及肿胀率，并做统计分析。
肿胀度＝炎症组足趾重量-空白组足趾重量
结果如图2所示
DT002：青蒿素负载香菇菌多糖复合物；LPS：脂多糖；GK2：甘草酸二钾；Drag：红没药醇；Hydr：氢化可的松。(n＝3，**P<0.01)
结论：
DT002显著抑制角叉菜胶对小鼠足趾造成的肿胀，且效果显著优于阳性药甘草酸二钾和红没药醇，其抑炎效果略优于糖皮质激素氢化可的松。
实施例3.DT002对皮肤改善情况的检测
检测方法：
招募30名志愿者(其中15男，15女)，随机分为5组(对照、DT002、GK2、Drag、Hydr)，其中每组男女各3人。每次使用时先用湿巾或水洗将右手手腕内侧清理干净，随后护肤品均匀擦拭于右手手腕内侧部位，每天早晚各一次，连续使用4周共28天。分别于第0、7、14、28天使用DermaLab仪器重点检测并监测TEWL(经表皮水流失)、Hydration PN(水含量)Ultrasound(超声)、Elasticity(弹性)和Skin Color(肤色)共五项指标，每次检测时志愿者右手手腕内侧湿巾擦拭干净并晾晒20分钟左右至晾干后开始检测。仪器所在房间不通风，无阳光直射，室温保持在24-25℃，湿度保持在40％。最后对得到的数据做统计分析。
结果：
表1.对照组皮肤改善情况：
表2.DT002对皮肤的改善情况(DT002：青蒿素香菇菌多糖复合物)：
表3.甘草酸二钾对皮肤的改善情况(GK2：甘草酸二钾)：
表4.红没药醇对皮肤的改善情况(Drag：红没药醇)：
表5.氢化可的松对皮肤的改善情况(Hydr：氢化可的松)：
结论：
DT002可有效改善皮肤的保水性能，提高皮层胶原蛋白含量，改善皮肤弹性。对保水性能的最高改善率、对胶原蛋白含量的改善以及对皮肤弹性的改善均优于三种阳性药。与对照组比较，DT002对皮肤的改善效果优于甘草酸二钾、红没药醇和氢化可的松。
尽管已经示出和描述了本发明的实施例，对于本领域的普通技术人员而言，可以理解在不脱离本发明的原理和精神的情况下可以对这些实施例进行多种变化、修改、替换和变型，本发明的范围由所附权利要求及其等同物限定。
Claims (9)
1.青蒿素负载香菇菌多糖复合物，其特征在于：包括按重量百分含量组分的原料：
香菇菌多糖，含量80-99％；
青蒿素，含量1％-20％。
2.根据权利要求1所述的青蒿素负载香菇菌多糖复合物，其特征在于：所述香菇菌多糖，又称香菇多糖，Lentinan，CAS:37339-90-5，分子量：1152.9995，灰白色粉末；所述青蒿素，又称黄华素，Artemisinine，CAS:63968-64-9，分子量：282.33，无色针状晶体。
3.权利要求1所述的青蒿素负载香菇菌多糖复合物的制备方法，其特征在于，所述香菇菌多糖采用生物发酵后提取方式制备，然后与青蒿素通过化学合成方式获得青蒿素负载香菇菌多糖复合物复合物，包括以下步骤：
步骤一：以新鲜绿茶茶叶为主，蔗糖、蛋白胨、磷酸氢钾等为辅，按照茶叶与水重量比为1:1的比例使茶叶和水充分混匀；在121℃高温条件下充分灭菌20-30分钟后得到茶叶培养基；将所述茶叶培养基与香菇菌丝体共培养；对香菇菌丝体的发酵产物水提醇沉分离提取，最终精制得到香菇菌多糖；
步骤二：香菇菌多糖与青蒿素以相应的比例混合在去离子水中；加入与青蒿素等当量的二环己基碳二亚胺作为缩合剂；在室温下搅拌反应24h；透析24小时，除去多余二环己基碳二亚胺以及未反应的青蒿素；冷冻干燥，最终得到相应的青蒿素负载香菇菌多糖复合物。
4.根据权利要求3所述的制备方法，其特征在于：所述香菇菌丝体采用的香菇菌品种不限于某单一品种的香菇菌。
5.根据权利要求1所述的青蒿素负载香菇菌多糖复合物，其特征在于：主要以水性糊剂或凝胶或液体的形式，优选为水性液体的形式。
6.根据权利要求5所述的青蒿素负载香菇菌多糖复合物，其特征在于：将青蒿素负载香菇菌多糖复合物配制为含水洗液、油包水或水包油乳液、油或油.醇洗液、阴离子或非离子两亲脂质的囊状分散体、含水凝胶、水.醇凝胶、醇凝胶或油.醇凝胶、固体棒或气溶胶。
7.一种青蒿素负载香菇菌多糖复合物用于抗炎、抗氧化衰老、护肤的制剂。
8.根据权利要求7所述的应用，其特征在于：所述剂型为临床上可接受的任一剂型。
9.根据权利要求7所述的应用，其特征在于：所述青蒿素负载香菇菌多糖复合物为所述抗炎、抗氧化衰老或护肤的唯一活性成分，或该复合物与其他物质一起制备所述抗炎、抗氧化衰老或护肤的制剂。
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