CN105999232B - 包含升麻皂苷的药物组合物及其应用 - Google Patents
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Abstract
本发明涉及一种包含升麻皂苷的药物组合物及其应用，属于中药领域。其由升麻皂苷提取物和降钙素原组成。所述升麻皂苷提取物和降钙素原的重量比为1∶0.01‑1。
Description
技术领域
本发明涉及一种包含升麻皂苷的药物组合物及其应用，属于中药领域。
背景技术
升麻是我国著名的常用中药，地下根茎入药，其药用功效为清热解毒、升举阳气和发表透疹。民间主治头痛、牙痛、咽喉痛、麻疹及子宫脱垂等妇科疾病。在我国，升麻的药用价值早在《神农本草经》中就有记载，并被列为“上品”药材。在国内升麻属植物：绿升麻(Cimicifuga foetida)、兴安升麻(C dahurica)、大三叶升麻(C heracleifolia)的根茎均作为中药升麻载入《中华人民共和国药典》2005版。同属植物黑升麻(C racemosa)，又称蛇根草，在欧美国家也有悠久的使用历史。在美国，黑升麻根茎的醇提物用来治疗绝经期妇女的潮热和其它症状已有一百多年的历史。在欧洲，黑升麻作为天然雌激素替代品，治疗妇产科疾病也已有50年的历史。近十年来美国黑升麻干燥根茎的醇提取物(含总三萜皂苷成分32.5％)作为缓解更年期综合症的植物食品补充剂交易量达每年10亿美元左右，一直是十种最受欢迎的产品之一。在德国，黑升麻根茎的醇提物也被推荐来治疗经前综合症，经期腹痛和更年期综合症等症状。升麻属植物的化学成分和药理活性研究一直是国际上的研究热点。对升麻中化学成分的研究主要集中在三萜及其苷类成份上，对其药理活性研究则以类雌激素、抗骨质疏松为主。
肿瘤是一类严重威胁人类健康的致死性疾病，其发病机制尚未完全阐明，治疗效果亦不尽人意。目前常规用于肿瘤治疗的大部分药物，均是针对细胞增殖过程中DNA复制或细胞骨架的调控来设计的。这些药物在对快速增殖的肿瘤细胞进行杀伤的同时，对人体功能维持所需的正常增殖细胞(如骨髓干细胞等等)也有很大的毒性，导致接受治疗的病人机体正常功能严重受损，对于肿瘤的抵抗能力反而下降，难以取到很好的治疗效果。
在众多肿瘤当中，乳腺癌是女性最常见的癌症原因，并且在美国是女性第二大常见的癌症死亡原因。虽然多数初期乳腺癌是基于发现乳腺影像异常来诊断的，但是肿块或乳腺组织坚实度的变化也可能是疾病的警报信号。过去几十年内提高乳腺癌风险意识已致使进行乳腺摄影筛查的女性数量增加，导致更早期检测癌症并且由此提高存活率。尽管如此，乳腺癌是45至55岁女性最常见的死亡原因。
降钙素原(procalcitonin，PCT)是一个有113个氨基酸的无激素活性的降钙素前肽物质，分子量约为13kD。当严重细菌、真菌、寄生虫感染以及脓毒症和多脏器功能衰竭时它在血浆中的水平升高。自身免疫、过敏和病毒感染时PCT不会升高。局部有限的细菌感染、轻微的感染和慢性炎症不会导致其升高。因此其在细菌感染检测中起到了至关重要的作用。随着研究的深入，有研究表明PCT中可能存在一些CTL表位肽，将这些CTL表位肽分离制备应用其可能通过诱导体内T细胞而表现出一定的抗肿瘤作用，但是PCT是否具有抗肿瘤作用本领域目前没有定论。
发明内容
本发明的一个目的是提供一种可用于治疗乳腺癌的药物组合物，其由升麻皂苷提取物和降钙素原组成。
在一个实施方案中，所述升麻皂苷提取物和降钙素原的重量比为1∶0.01-1；优选为1∶0.15。
在另一个实施方案中，所述升麻皂苷提取物可通过市售获得但也可以通过本领域的常规方法进行制备，例如，将升麻(根)药材粉碎过筛，以10倍量的80％乙醇加热回流提取3次，每次1h，提取完毕后过滤，滤液浓缩成浸膏，用大孔树脂D101进行柱层析，先用正已烷洗脱油脂，然后用乙酸乙酯洗脱，洗脱液浓缩干燥得升麻皂苷提取物。
在进一步地实施方案中，所述药物组合物由升麻皂苷提取物、降钙素原和阿维A组成，升麻皂苷提取物、降钙素原和阿维A的重量比为1∶0.01-1∶0.01-1；优选为1∶0.15∶0.07。
本发明的另一个目的是提供所述药物组合物在制备治疗肿瘤疾病药物中的应用。
在一个实施方案中，所述肿瘤疾病为乳腺癌。
具体实施方式
还可进一步通过实施例来理解本发明，其中所述实施例说明了一些制备或使用方法。然而，要理解的是，这些实施例不限制本发明。现在已知的或进一步开发的本发明的变化被认为落入本文中描述的和以下要求保护的本发明范围之内。
实施例1升麻皂苷提取物的制备
将升麻(根)药材10kg粉碎过筛，以10倍量的80％乙醇加热回流提取3次，每次1h，提取完毕后过滤，滤液浓缩成浸膏，用大孔树脂D101进行柱层析，先用正已烷洗脱油脂，然后用乙酸乙酯洗脱，洗脱液浓缩干燥得升麻皂苷提取物362g。
实施例2药物组合物对肿瘤细胞系的增殖抑制作用
取人乳腺癌细胞系(MCF-7)、人宫颈癌细胞系(Hela)以及人卵巢癌细胞系(skov3)采用含10％FBS的RPMI1640培养基传代培养，扩增后收获并接种到96孔板，密度为10000个/孔。培养1天后加入含药物的培养基，对照组添加等量的培养基，在孵箱中继续培养48小时，加MTT，采用酶标仪测定吸光度(OD590)，以对照组吸光度为100％计算各药物组抑制率。
各药物组药物组成及重量比
具体结果如下：
| 抑制率(％) | MCF-7 | Hela | skov3 |
| 组1 | 10.3±1.1 | 14.2±1.3 | 7.4±1.1 |
| 组2 | 25.2±1.2 | 28.5±1.6 | 17.3±1.3 |
| 组3 | 1.3±0.3 | 1.7±0.4 | 0.5±0.2 |
| 组4 | 3.6±0.5 | 4.4±0.5 | 1.1±0.3 |
| 组5 | 36.4±1.6 | 30.2±1.8 | 18.4±0.9 |
| 组6 | 48.9±2.1 | 31.7±1.9 | 19.5±1.1 |
| 组7 | 34.3±1.9 | 29.9±1.5 | 18.9±1.2 |
| 组8 | 55.1±2.3 | 32.6±2.1 | 19.2±0.9 |
有趣的是，我们在乳腺癌细胞系MCF-7中发现了升麻皂苷提取物、降钙素原和/或阿维A的抑制协同作用，而在其他女性相关肿瘤细胞系中上述药物组合物却没有体现出明显的协同作用，因此我们进一步地在多个乳腺癌细胞系中进行了上述测试，结果表明上述药物组合物在乳腺癌细胞系Hs-578T、BT-549、MDA-MB-436以及HCC1937中均表现出了明显的协同抑制作用。
实施例3中药组合物对小鼠乳腺癌模型的影响
取ICR小鼠，将人乳腺癌细胞(MCF-7)接种于小鼠的背侧腋窝皮下，等瘤体长至1-1.5cm时，选择良好的瘤组织，制备成单细胞悬液，取0.2ml该悬液(癌细胞数为1×106个/ml)背侧腋窝皮下接种。接种24小时后给药，连续灌胃给药14次，每天一次。末次给药后，处死小鼠计算抑瘤率，抑瘤率＝(模型组瘤重-给药组瘤重)/模型组瘤重×100％。
具体结果如下：
进一步地，本发明对各组小鼠收集的MCF-7瘤组织，制备成MCF-7细胞悬液，并采用流式细胞仪检测了各组MCF-7细胞的表面CD47的表达情况，有研究表明肿瘤细胞表面均存在高表达的CD47，而CD47可以和其配体SIRP结合构成拒吞噬信号(do not eat me)，从而防止巨噬细胞对肿瘤细胞识别和吞噬，因此检测肿瘤细胞表面的CD47表达是非常重要的。
具体结果如下：
本发明内容仅仅举例说明了要求保护的一些具体实施方案，其中一个或更多个技术方案中所记载的技术特征可以与任意的一个或多个技术方案相组合，这些经组合而得到的技术方案也在本申请保护范围内，就像这些经组合而得到的技术方案已经在本发明公开内容中具体记载一样。
Claims (3)
1.一种用于治疗乳腺癌的药物组合物，其由升麻皂苷提取物和降钙素原组成；所述升麻皂苷提取物和降钙素原的重量比为1∶0.15。
2.一种用于治疗乳腺癌的药物组合物，其特征在于，所述药物组合物由升麻皂苷提取物、降钙素原和阿维A组成，升麻皂苷提取物、降钙素原和阿维A的重量比为1∶0.15∶0.07。
3.权利要求1或2任一项所述药物组合物在制备治疗肿瘤疾病药物中的应用，所述肿瘤疾病为乳腺癌。
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