CN115531457A - 一种具有抗肿瘤功能的中药组合物及其制备方法 - Google Patents
一种具有抗肿瘤功能的中药组合物及其制备方法 Download PDFInfo
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Abstract
本发明属于中药技术领域,具体涉及一种具有抗肿瘤作用的中药组合物。所述中药组合物主要由羌活、独活、茯苓、防风、荆芥、川芎、桔梗、柴胡、前胡、枳壳、甘草制备而成。本发明中药组合物,临床使用安全性高。本发明试验发现本发明中药组合物对于多种肿瘤具有显著的抑制作用,本发明组合物新用途的发现具有广阔的市场前景。
Description
技术领域
本发明涉及一种中药组合物及其制备方法,具体涉及一种具有抗肿瘤功能的中药组合物及其制备方法,属于中药技术领域。
背景技术
肿瘤是指机体在各种致瘤因子作用下,局部组织细胞增生所形成的新生物。肿瘤分为良性和恶性两大类。良性肿瘤绝大多数不会恶变,但需要密切关注,防止其恶化。恶行肿瘤,也就是通常人们所说的“癌症”,已成为威胁人类健康,致人死亡的一大病因。放化疗是肿瘤治疗的常用方案,但是放射线和化疗药物在杀伤肿瘤细胞的同时,对正常细胞造成一定程度的损伤。
近年中药在肿瘤临床和康复治疗中发挥越来越大的作用,不仅可以直接抑制肿瘤细胞的增长,而且还能增强患者的免疫力防治肿瘤转移和复发。口服中药是目前中医治疗肿瘤最常用的方法,胃黏膜吸收为最主要吸收途径,但有些抗肿瘤中药有一定的刺激作用,会对胃黏膜造成一定的损伤,所以病人用药后可出现恶心、呕吐、胃痉挛疼痛、胃黏膜出血等副作用,限制了有些口服抗肿瘤药物的临床使用。另外,多数中药对肿瘤的针对作用不强,只能作为辅助治疗药物。
荆防败毒散是由人参败毒散去人参加荆芥、防风而成。人参败毒散最早出自宋代官方成药医籍《太平惠民和剂局方》,药物组成为柴胡、甘草、桔梗、人参、川芎、茯苓、枳壳、前胡、羌活、独活,长于治疗疫病。原书记载:“治伤寒时气。”《类证活人书》和《小儿药证直诀》均言其“治瘟疫……瘟疫时行……此药不可缺也。”明末清初医家喻昌对此方十分推崇,指出:“人感三气两病,病而死,其气互传,乃至十百千万,传为疫矣。倘病者日服此药二三剂,所受疫邪,不复留于胸中,讵不快哉。”还称赞:“昌鄙见三气门中,推此方为第一,以其功之著也。”提出人参败毒散诸药辛平,乃“治疫第一方”,可称为“活人败毒散”。
荆防颗粒是荆防败毒散的中成药制剂。荆防(冲剂)颗粒是《卫生部药品标准》中药成方制剂第二册收载的品种,标准编号为WS3-B-0328-90。王乖娟等(王乖娟,刘卫兵,荆鲁华.荆防颗粒治疗血虚风燥型皮肤瘙痒症临床观察[J].中国中西医结合皮肤性病学杂志,2009,8(05):315)自拟荆防颗粒治疗血虚风燥型皮肤瘙痒症,实验发现荆防颗粒治疗血虚风燥型皮肤瘙痒症远期临床疗效满意。谢舜辉等(谢舜辉,郑慕雄,陈昌鹏,陈玉兴.荆防颗粒对大鼠Ⅰ型超敏反应及肥大细胞释放组胺的影响[J].皮肤性病诊疗学杂,2010,17(01):21-23)研究发现荆防颗粒能明显抑制大鼠Ⅰ型超敏反应和肥大细胞释放组胺。周小花等(周小花,刘冬梅,兰燕琴,胡毅翔,俞文英,余陈欢.荆防颗粒治疗扁平疣的临床疗效及作用机制研究[J].中华中医药学刊,2015,33(10):2374-2377)研究发现荆防颗粒内服治疗扁平疣具有一定的临床疗效和安全性。中国专利CN201410174688.5公开了一种荆防颗粒的检测方法。
发明内容
本发明的主要目的在于提供一种具有抗肿瘤作用的中药组合物,所述的中药组合物主要由羌活、独活、茯苓、防风、荆芥、川芎、桔梗、柴胡、前胡、枳壳、甘草制备而成。
本发明是在现有产品“荆防颗粒”的基础上所做的对其用途的进一步开发。
荆芥、防风辛温解表,祛风散寒为主药。羌活、独活祛风除湿止痛,且助荆芥、防风解表之力;川芎祛血中之风而宣痹止痛,共为辅药。柴胡辛凉透表,可疏散肌表郁热,并与前胡、桔梗、枳壳共用有升有降,共起宣肺化痰、开胸利气之功效;茯苓甘淡而平,可健脾渗湿,此4味共为佐药。甘草为使,调和诸药。诸药合用,共奏发汗解表、祛风散寒除湿之功。它具有发汗解表,散风祛湿的作用。临床上用于治疗感冒风寒,头痛身痛,恶寒无汗,鼻塞流涕,咳嗽。是目前市场上用于治疗感冒风寒,头痛身痛,恶寒无汗,鼻塞流涕,咳嗽的常用药品。
肿瘤结节更易光顾寒性体质,癌症患者有一多半都是寒性体质:第一,寒性体质的人一般血液流动较慢,易形成瘀血和痰浊、气滞相互“勾结”的状况,“痰瘀互结”积滞在某个部位,长期在毒邪的作用下,就容易导致肿瘤的形成。第二,寒性体质的人不能有效阻挡外部毒性物质的侵入,清除毒素的能力也相对较差,为肿瘤产生创造了先决条件。
荆防颗粒功效为散风祛湿、发汗解表除寒,可以祛除寒湿,改善寒湿体质,消除肿瘤发生发展的体质因素。
基于此,发明人进行了荆防颗粒抗肿瘤活性的研究,发现荆防颗粒对多种肿瘤类型(肺癌、肝癌、胃癌、结直肠癌、食管癌、膀胱癌、胰腺癌、脑瘤、乳腺癌、甲状腺癌、淋巴瘤、子宫颈癌或子宫体癌)具有显著的治疗效果。并且本发明中药组合物对于正常细胞没有抑制作用,安全性高。
药效实施例1研究发现本发明中药组合物组灌胃给药后,分离的动物血清对U87、MGC80-3、CaES-17、MCF-7、Hela、Ishikawa、CEM/C1、PANC-1、K562均有抑制作用。
药效实施例2表明本发明中药组合物能够显著改善小鼠黑色素瘤肺转移状况。
药效实施例3表明本发明中药组合物对H22肝癌抑瘤率高。本发明中药组合物具有较好的体内抗动物移植性肿瘤的作用。
药效实施例4表明本发明中药组合物对CT26.WT结肠癌具有显著的治疗作用。
本发明目的之一在于,提供上述中药组合物的组成,即所述中药组合物主要由以下原料制备而成:
荆芥5-30重量份 防风5-30重量份 羌活5-30重量份 独活5-30重量份 柴胡3-25重量份 前胡3-25重量份 川芎5-30重量份 枳壳3-25重量份 茯苓5-30重量份 桔梗3-25重量份 甘草1-10重量份。
进一步地,所述中药组合物主要由以下原料制备而成:
荆芥10-20重量份 防风10-20重量份 羌活10-20重量份 独活10-20重量份 柴胡3-20重量份 前胡3-20重量份 川芎10-20重量份 枳壳3-20重量份 茯苓10-25重量份 桔梗3-20重量份 甘草1-8重量份。
更进一步地,所述中药组合物主要由以下原料制备而成:
荆芥15重量份 防风15重量份 羌活15重量份 独活15重量份 柴胡15重量份 前胡15重量份 川芎15重量份 枳壳15重量份 茯苓15重量份 桔梗15重量份 甘草5重量份。
更进一步地,所述中药组合物主要由以下原料制备而成:
荆芥10重量份 防风10重量份 羌活12重量份 独活12重量份 柴胡5重量份 前胡5重量份 川芎12重量份 枳壳5重量份 茯苓20重量份 桔梗5重量份 甘草3重量份。
本发明目的之二在于提供上述中药组合物的具体用途,主要有:
上述中药组合物在制备抗肿瘤药物中的用途。
在一个实施方式中,所述肿瘤为头颈部肿瘤、胸部肿瘤、消化系统肿瘤、泌尿生殖系统肿瘤或淋巴及血液系统肿瘤中的一种或几种。
在一个实施方式中,所述头颈部肿瘤为鼻腔及鼻窦恶性肿瘤、鼻咽癌、口腔癌、喉癌、涎腺肿瘤、颅内肿瘤、甲状腺癌或舌癌。
在一个实施方式中,所述胸部肿瘤为肺癌、食管癌、贲门癌、乳腺癌、或纵膈肿瘤。
在一个实施方式中,所述消化系统肿瘤为胃癌、大肠癌及乙状结肠和直肠癌、肝癌、胰腺癌与壶腹周围癌、胆道肿瘤或小肠恶性肿瘤。
在一个实施方式中,所述泌尿生殖系统肿瘤为肾癌、膀胱癌、睾丸恶性肿瘤、阴茎癌、子宫颈癌、子宫内膜癌或卵巢癌。
在一个实施方式中,所述淋巴及血液系统肿瘤为淋巴瘤、多发性骨髓瘤、急性白血病、慢性白血病、红细胞增多症、血小板增多症、骨髓增生异常综合症、骨髓纤维化或恶性组织细胞病。
在一个实施方式中,所述肿瘤为实体瘤、淋巴瘤或白血病。
在一个实施方式中,所述肿瘤为肺癌、肝癌、胃癌、结直肠癌、食管癌、前列腺癌、膀胱癌、胰腺癌、脑瘤、乳腺癌、甲状腺癌、淋巴瘤、子宫颈癌或子宫体癌中的一种或几种。
本发明目的之三在于提供上述中药组合物的制备方法,所述制备方法主要包括以下步骤:
步骤A:将荆芥、防风、羌活、独活、前胡、川芎、枳壳,蒸馏提取挥发油备用,蒸馏后的药渣、蒸馏后的川芎、枳壳水溶液备用;
步骤B:将步骤A所得蒸馏后的川芎、枳壳水溶液配制成10-40%的乙醇溶液,备用;
步骤C:将茯苓、步骤A所得蒸馏后的川芎、枳壳药渣混合,以步骤B所得乙醇溶液渗漉提取,渗漉液备用;
步骤D:将柴胡、桔梗、甘草、步骤A所得蒸馏后的荆芥、防风、羌活、独活、前胡药渣,加水煎煮,煎煮液浓缩后备用;
步骤E:将步骤C所得渗漉液、步骤D所得煎煮液混合,浓缩,加入步骤A所得挥发油,即得。
进一步地,所述制备方法主要包括以下步骤:
步骤A:将荆芥、防风、羌活、独活、前胡、川芎、枳壳,蒸馏提取挥发油备用,蒸馏后的药渣、蒸馏后的川芎、枳壳水溶液备用;
步骤B:将步骤A所得蒸馏后的川芎、枳壳水溶液配制成15-30%的乙醇溶液,备用;
步骤C:将茯苓、步骤A所得蒸馏后的川芎、枳壳药渣混合,以步骤B所得乙醇溶液渗漉提取,渗漉液备用;
步骤D:将柴胡、桔梗、甘草、步骤A所得蒸馏后的荆芥、防风、羌活、独活、前胡药渣,加水煎煮,煎煮液浓缩成稠膏,备用;
步骤E:将步骤C所得渗漉液、步骤D所得稠膏混合,浓缩成清膏,加入步骤A所得挥发油,即得。
更进一步地,所述制备方法主要包括以下步骤:
步骤A:将荆芥、防风、羌活、独活、前胡、川芎、枳壳,蒸馏提取挥发油备用,蒸馏后的药渣、蒸馏后的川芎、枳壳水溶液备用;
步骤B:将步骤A所得蒸馏后的川芎、枳壳水溶液配制成25%的乙醇溶液,备用;
步骤C:将茯苓、步骤A所得蒸馏后的川芎、枳壳药渣混合,以步骤B所得乙醇溶液渗漉提取,渗漉液备用;
步骤D:将柴胡、桔梗、甘草、步骤A所得蒸馏后的荆芥、防风、羌活、独活、前胡药渣,加水煎煮,煎煮液浓缩成稠膏,备用;
步骤E:将步骤C所得渗漉液、步骤D所得稠膏混合,浓缩成清膏,加入步骤A所得挥发油,即得。
本发明目的之四在于提供含有上述中药组合物的中药制剂,所述的中药制剂为临床上可接受的口服制剂;
进一步地,所述临床上可接受的口服制剂为丸剂、胶囊剂、片剂、颗粒剂或液体口服制剂中的一种或多种;
更进一步地,所述口服制剂为颗粒剂。
本发明中药组合物用于抗肿瘤时,按生药计用量为0.02g/kg·d~3.0g/kg·d。优选地,所述中药组合物按生药计用量为0.05g/kg·d~1.4g/kg·d。进一步优选地,所述中药组合物按生药计用量为0.29g/kg·d。
与现有技术相比,本发明取得了显著的技术效果:
本发明中药组合物,临床使用安全性高。本发明试验发现本发明中药组合物对于多种肿瘤具有显著的抑制作用,本发明组合物新用途的发现具有广阔的市场前景。
具体实施方式
为验证本发明中药组合物抗肿瘤方面的功效,发明人开展了相关的药效学试验研究。需要说明的是,下述药效学试验所选取的药品为本发明具有代表性的配方及其制备方法所得的药品;本发明所包含的其它配方及制备方法所得药品,发明人同样进行了药效学实验,实验结果显示其他配方及制备方法所得药品具有相同或类似的效果,但由于篇幅限制,在此不一一穷举;此外,下述药效学实验仅以体外实验和部分代表性的动物模型为例验证本发明的功效。
发明人要说明的是,以下实验研究均是在急性毒性试验、长期毒性试验证明药物安全性基础之上开展,实验研究中的给药剂量均在安全剂量范围之内。
制备实施例1颗粒剂制备
处方:
制备方法:
步骤A:将荆芥、防风、羌活、独活、前胡、川芎、枳壳,分别蒸馏提取挥发油备用,蒸馏后的药渣、蒸馏后的川芎、枳壳水溶液备用;
步骤B:将步骤A所得蒸馏后的川芎、枳壳水溶液配制成25%的乙醇溶液,备用;
步骤C:将茯苓、步骤A所得蒸馏后的川芎、枳壳药渣混合,以步骤B所得乙醇溶液渗漉提取,渗漉液备用;
步骤D:将柴胡、桔梗、甘草、步骤A所得蒸馏后的荆芥、防风、羌活、独活、前胡药渣,加水煎煮两次,每次1.5小时,合并两次的煎煮液,过滤后浓缩成稠膏,备用;
步骤E:将步骤C所得渗漉液、步骤D所得稠膏混合,静置,过滤后浓缩成清膏,加蔗糖适量,混匀,制成颗粒,干燥,加入步骤A所得挥发油,混匀,即得。
制备实施例2颗粒剂制备
处方:
制备方法同实施例1。
制备实施例3颗粒剂制备
处方:
制备方法同实施例1。
制备实施例4口服液制备
处方:
制备方法:
步骤A:将荆芥、防风、羌活、独活、前胡、川芎、枳壳,分别蒸馏提取挥发油备用,蒸馏后的药渣、蒸馏后的川芎、枳壳水溶液备用;
步骤B:将步骤A所得蒸馏后的川芎、枳壳水溶液配制成10%的乙醇溶液,备用;
步骤C:将茯苓、步骤A所得蒸馏后的川芎、枳壳药渣混合,以步骤B所得乙醇溶液渗漉提取,渗漉液备用;
步骤D:将柴胡、桔梗、甘草、步骤A所得蒸馏后的荆芥、防风、羌活、独活、前胡药渣,加水煎煮两次,每次1.5小时,合并两次的煎煮液,过滤后浓缩成稠膏,备用;
步骤E:将步骤C所得渗漉液、步骤D所得稠膏混合,静置,过滤后浓缩成清膏,加蔗糖适量,混匀,加入步骤A所得挥发油,混匀,加水至1000ml,即得。
制备实施例5糖浆剂制备
处方:
制备方法:
步骤A:将荆芥、防风、羌活、独活、前胡、川芎、枳壳,分别蒸馏提取挥发油备用,蒸馏后的药渣、蒸馏后的川芎、枳壳水溶液备用;
步骤B:将步骤A所得蒸馏后的川芎、枳壳水溶液配制成40%的乙醇溶液,备用;
步骤C:将茯苓、步骤A所得蒸馏后的川芎、枳壳药渣混合,以步骤B所得乙醇溶液渗漉提取,渗漉液备用;
步骤D:将柴胡、桔梗、甘草、步骤A所得蒸馏后的荆芥、防风、羌活、独活、前胡药渣,加水煎煮两次,每次1.5小时,合并两次的煎煮液,过滤后浓缩成稠膏,备用;
步骤E:将步骤C所得渗漉液、步骤D所得稠膏混合,静置,过滤后浓缩成清膏,加蔗糖适量,混匀,加入步骤A所得挥发油和单糖浆500ml,混匀,静置,滤过,加水至1000ml,即得。
制备实施例6片剂制备
处方:
制备方法:
步骤A:将荆芥、防风、羌活、独活、前胡、川芎、枳壳,分别蒸馏提取挥发油备用,蒸馏后的药渣、蒸馏后的川芎、枳壳水溶液备用;
步骤B:将步骤A所得蒸馏后的川芎、枳壳水溶液配制成15%的乙醇溶液,备用;
步骤C:将茯苓、步骤A所得蒸馏后的川芎、枳壳药渣混合,以步骤B所得乙醇溶液渗漉提取,渗漉液备用;
步骤D:将柴胡、桔梗、甘草、步骤A所得蒸馏后的荆芥、防风、羌活、独活、前胡药渣,加水煎煮两次,每次1.5小时,合并两次的煎煮液,过滤后浓缩成稠膏,备用;
步骤E:将步骤C所得渗漉液、步骤D所得稠膏混合,静置,过滤后浓缩成清膏,加蔗糖适量,混匀,制成颗粒,干燥,加入步骤A所得挥发油,混匀,制成颗粒,加入辅料适量,混匀,压片,即得。
制备实施例7胶囊剂制备
处方:
步骤A:将荆芥、防风、羌活、独活、前胡、川芎、枳壳,分别蒸馏提取挥发油备用,蒸馏后的药渣、蒸馏后的川芎、枳壳水溶液备用;
步骤B:将步骤A所得蒸馏后的川芎、枳壳水溶液配制成30%的乙醇溶液,备用;
步骤C:将茯苓、步骤A所得蒸馏后的川芎、枳壳药渣混合,以步骤B所得乙醇溶液渗漉提取,渗漉液备用;
步骤D:将柴胡、桔梗、甘草、步骤A所得蒸馏后的荆芥、防风、羌活、独活、前胡药渣,加水煎煮两次,每次1.5小时,合并两次的煎煮液,过滤后浓缩成稠膏,备用;
步骤E:将步骤C所得渗漉液、步骤D所得稠膏混合,静置,过滤后浓缩成清膏,加蔗糖适量,混匀,制成颗粒,干燥,加入步骤A所得挥发油,混匀,制成颗粒,干燥,粉碎,装入胶囊,即得。
制备实施例8丸剂制备
处方:
制备方法:
步骤A:将荆芥、防风、羌活、独活、前胡、川芎、枳壳,分别蒸馏提取挥发油备用,蒸馏后的药渣、蒸馏后的川芎、枳壳水溶液备用;
步骤B:将步骤A所得蒸馏后的川芎、枳壳水溶液配制成25%的乙醇溶液,备用;
步骤C:将茯苓、步骤A所得蒸馏后的川芎、枳壳药渣混合,以步骤B所得乙醇溶液渗漉提取,渗漉液备用;
步骤D:将柴胡、桔梗、甘草、步骤A所得蒸馏后的荆芥、防风、羌活、独活、前胡药渣,加水煎煮两次,每次1.5小时,合并两次的煎煮液,过滤后浓缩成稠膏,备用;
步骤E:将步骤C所得渗漉液、步骤D所得稠膏混合,静置,过滤后浓缩成清膏,加蔗糖适量,混匀,制成颗粒,干燥,加入步骤A所得挥发油,混匀,干燥,粉碎,过筛,加炼蜜40~60g,适量的水泛丸,干燥,即得。
制备实施例9浸膏制备
处方:
制备方法:
步骤A:将荆芥、防风、羌活、独活、前胡、川芎、枳壳,分别蒸馏提取挥发油备用,蒸馏后的药渣、蒸馏后的川芎、枳壳水溶液备用;
步骤B:将步骤A所得蒸馏后的川芎、枳壳水溶液配制成25%的乙醇溶液,备用;
步骤C:将茯苓、步骤A所得蒸馏后的川芎、枳壳药渣混合,以步骤B所得乙醇溶液渗漉提取,渗漉液备用;
步骤D:将柴胡、桔梗、甘草、步骤A所得蒸馏后的荆芥、防风、羌活、独活、前胡药渣,加水煎煮两次,每次1.5小时,合并两次的煎煮液,过滤后浓缩成稠膏,备用;
步骤E:将步骤C所得渗漉液、步骤D所得稠膏混合,静置,过滤后浓缩成清膏,加入步骤A所得挥发油,混匀,即得。
药效实施例1本发明中药组合物的体外抗肿瘤试验
1.实验方法
1.1含药血清的制备
取小鼠72只,体重22~26g,雌雄各半。
分为4组:雌性正常对照组,雄性正常对照组,雌性给药组、雄性给药组。
正常对照组,ig蒸馏水;
给药组:给药实施例9中的浸膏,按生药计0.97g/kg。
每天3次,连续ig 3d,末次给药或蒸馏水后1h,戊巴比妥钠麻醉后,下腔静脉取血,分离血清,56℃灭活30min,-20℃保存备用。
实验分组:
高剂量组:血清终浓度为40%;低剂量组:血清终浓度为20%;
正常对照组:血清终浓度为40%或20%;
空白对照组:不含血清,加等量培养液。
实验细胞株:人脑瘤细胞U87、人胃癌细胞MGC80-3、人食道癌细胞CaES-17、人乳腺癌细胞MCF-7、人宫颈癌细胞Hela、人子宫内膜癌Ishikawa、人急性淋巴细胞白血病细胞CEM/C1、人胰腺癌细胞PANC-1、人慢性髓系白血病细胞K562、人肾皮质近曲小管上皮细胞HK-2、人正常胃粘膜细胞GES-1。
1.2.含药血清对细胞株的影响
取对数生长期细胞,调整密度为3~5×104个/ml,每孔100μl,接种于96孔板,4h后加入含药血清使终浓度分别为40%或20%,使其总体积为200μl。每个浓度6个复孔。用同浓度正常组血清作为正常对照,加等量培养液作为空白对照。
处理48h,然后加入CCK-8溶液,20μl/孔。继续培养2h。在450nm波长处用酶标仪测A值。作组间t检验,并按以下公式计算抑制率,抑制率%=【正常对照组A均数-空白对照组A均数)-(含药血清组A均数-空白对照组A均数)】/(正常对照组A均数-空白对照组A均数)×100%。
1.3.统计方法
所有实验数据均以表示,采用SPSS17.0数据处理软件进行方差分析和组间t检验。P<0.05有统计学意义。
2.结果
2.1实验结果
本发明中药组合物组ig给药后,分离的动物血清对U87、MGC80-3、CaES-17、MCF-7、Hela、Ishikawa、CEM/C1、PANC-1、K562均有抑制作用。
本发明中药组合物对HK-2、GES-1没有抑制作用。
表1本发明中药组合物含药血清对细胞株的抑制率
注:-代表抑制率小于20%;
+代表抑制率大于20%;
++代表抑制率大于40%;
+++代表抑制率大于60%。
3.小结
实验结果表明,本发明中药组合物对U87、MGC80-3、CaES-17、MCF-7、Hela、Ishikawa、CEM/C1、PANC-1、K562体外培养细胞有一定的抑制作用。本发明中药组合物具有较好的体外抗肿瘤作用。本发明中药组合物用于抗肿瘤时安全性高。
实施例2本发明中药组合物对小鼠黑色素瘤肺转移的抑制作用
1.1实验动物及材料
SPF级KM小鼠,雄性,18-22g,40只。
黑色素瘤细胞B16-F10,购自上海细胞库,由鲁南制药集团新药药理中心传代保存。
1.2建立小鼠黑色素瘤肺癌转移模型
1.2.1体外培养小鼠黑色素瘤细胞B16-F10,生理盐水调节细胞3.3×106cell/ml,每只0.2
ml接种于实验小鼠尾静脉。
在接种前2天,开始连续给药。
1.2.2分组及给药
每组动物8只。
实施例9浸膏:0.96g/kg(低剂量)、2.87g/kg(中剂量)、8.60g/kg(低剂量);
除空白组和模型组灌胃给予生理盐水外,其余各组小鼠分别给予相应的药物。
1.3检测指标
对肺部黑色素瘤进行评分。
评分标准如下:正常饲养小鼠肺脏为0分;
肺部有10%为黑色,评分为1分;
肺部有20%为黑色,评分为2分;
肺部有30%为黑色,评分为3分;
肺部有40%为黑色,评分为4分;
肺部有50%为黑色,评分为5分;
肺部有60%为黑色,评分为6分;
肺部有70%为黑色,评分为7分;
肺部有80%为黑色,评分为8分;
肺部有90%为黑色,评分为9分;
肺部有95%为黑色,评分为10分。
1.4试验结果采用均值±标准差,组间比较采用t检验。
2.结果
表2本发明中药组合物对肺部黑色素瘤评分结果
#P和模型组相比P≤0.05,##P和模型组相比P≤0.01。
本发明中药组合物能够显著改善小鼠黑色素瘤肺转移状况。
实施例3本发明中药组合物对小鼠H22肝癌移植瘤的抑制作用
动物体内试验反映药物的疗效与毒性的综合结果,是体外检测方法与模型不能取代的药物筛选过程的决定性环节。移植肿瘤模型是目前研究最多的肿瘤模型,容易人为操作,动物成瘤稳定,均一性好,发病率高,成瘤时间差异不大,容易施加干扰因素。体内试验通常采用动物肿瘤移植模型和人癌异体移植瘤模型。
1.1实验动物及材料
SPF级KM小鼠,雄性,18-22g,32只。
H22肝癌瘤株,购自武汉大学保藏中心,由鲁南制药集团新药药理中心传代保存。
1.2建立H22肝癌荷瘤小鼠模型
从液氮中取出冻存的H22瘤株,投入37℃水浴中,迅速摇晃使其快速溶化,移入超净台,用生理盐水洗1遍,1000rpm,5min,弃上清,用生理盐水重悬,调细胞浓度为1×106ce1ls/ml,接种到小鼠腹腔,0.2m1/只,5-7天后传代,传3代,用1m1注射器无菌吸取传代H22瘤株且生长良好的小鼠腹水,用生理盐水稀释成2.5×106cells/m1,每鼠0.2ml接种于鼠右侧腋窝皮下建立荷瘤小鼠模型。
1.2.2分组及给药
每组动物8只。
实施例1颗粒剂:2.925g/kg(低剂量)、5.85g/kg(高剂量);
实施例2颗粒剂:5.85g/kg;
除模型组灌胃给予生理盐水外,其余各组小鼠分别给予相应的药物。
1.3检测指标:
1.3.1一般情况观察:观察给药后动物行为、反应、饮食及死亡情况,隔天记录体重。
1.3.2瘤重及抑瘤率(%):采血后,将小鼠脱颈处死,剥离肿瘤,分析天平称重,记录瘤重,并按以下公式计算抑瘤率(%)。
抑瘤率=(模型组瘤重-给药组瘤重)/模型组瘤重×100%。
1.3.3对免疫器官的影响
取胸腺、脾脏,分析天平称重,记录脏器重,并按以下公式计算脏器指数。
脏器系数=脏器重量(mg)/体重(10g)。
1.4试验结果采用均值±标准差,组间比较采用t检验。
2.结果
2.1一般情况观察结果
由下表可知,与模型组比较,本发明中药组合物组体重均明显降低,具有统计学意义。
与模型组比较,本发明中药组合物组降低瘤重明显降低,具有统计学意义。
表3本发明化合物对H22肝癌移植瘤小鼠体重、瘤重、抑瘤率的影响
#P和模型组相比P≤0.05,##P和模型组相比P≤0.01。
2.2中药组合物对H22肝癌移植瘤小鼠脏器指数的影响
表4本发明化合物对H22肝癌移植瘤小鼠胸腺指数和脾脏指数的影响
#P和模型组相比P≤0.05,##P和模型组相比P≤0.01。
与模型组比较,本发明中药组合物组胸腺指数、脾脏指数均明显降低,具有统计学意义。
试验结果显示本发明中药组合物对H22肝癌抑瘤率高。本发明中药组合物具有较好的体内抗动物移植性肿瘤的作用。
实施例4本发明中药组合物对小鼠CT26.WT结肠癌移植瘤的抑制作用
1.1实验动物及材料
SPF级BALB/c小鼠,雄性,18-22g,32只。
CT26.WT结肠癌肿瘤株,购自中国科学院分子细胞科学卓越创新中心,由鲁南制药集团新药药理中心传代保存。
1.2建立CT26.WT结肠癌荷瘤小鼠模型
将CT26.WT细胞培养于含10%FBS的RPMI-1640培养基中,待细胞长至一定量时,用0.25%胰酶消化,吹散,用生理盐水悬浮,调整细胞浓度为5×106/ml,2.0ml/只接种于BALB/c小鼠右侧腋窝皮下,模型组瘤重达100~150mm3建模成功,按肿瘤体积分组开始给药,待模型组肿瘤体积达到预设体积(1000mm3)实验结束剖检动物。
1.2.2分组及给药
每组动物8只。
每组动物8只。
实施例1颗粒剂:2.925g/kg(低剂量)、5.85g/kg(高剂量);
实施例2颗粒剂:5.85g/kg;
除模型组灌胃给予生理盐水外,其余各组小鼠分别给予相应的药物。
1.3检测指标:
1.3.1一般情况观察:观察给药后动物行为、反应、饮食及死亡情况,隔天记录体重。
1.3.2瘤重及抑瘤率(%):采血后,将小鼠脱颈处死,剥离肿瘤,分析天平称重,记录瘤重,并按以下公式计算抑瘤率(%)。
抑瘤率=(模型组瘤重-给药组瘤重)/模型组瘤重×100%。
1.4试验结果采用均值±标准差,组间比较采用t检验。
2.结果
2.1一般情况观察结果
由下表可知,与模型组比较,本发明中药组合物组体重均明显降低,具有统计学意义。
与模型组比较,本发明中药组合物组降低瘤重明显降低,具有统计学意义。
表5本发明化合物对CT26.WT结肠癌移植瘤小鼠体重、瘤重、抑瘤率的影响
#P和模型组相比P≤0.05,##P和模型组相比P≤0.01。
试验结果显示本发明中药组合物对CT26.WT结肠癌具有显著的治疗作用。本发明中药组合物具有较好的体内抗动物移植性肿瘤的作用。
Claims (10)
1.一种主要由羌活、独活、茯苓、防风、荆芥、川芎、桔梗、柴胡、前胡、枳壳、甘草制备而成的中药组合物在制备抗肿瘤药物中的用途。
2.一种如权利要求1所述的用途,其特征在于,所述的肿瘤所述肿瘤为头颈部肿瘤、胸部肿瘤、消化系统肿瘤、泌尿生殖系统肿瘤或淋巴及血液系统肿瘤中的一种或几种。
3.如权利要求2所述的用途,其特征在于,所述头颈部肿瘤为鼻腔及鼻窦恶性肿瘤、鼻咽癌、口腔癌、喉癌、涎腺肿瘤、颅内肿瘤、甲状腺癌或舌癌;所述胸部肿瘤为肺癌、食管癌、贲门癌、乳腺癌、或纵膈肿瘤;所述消化系统肿瘤为胃癌、大肠癌及乙状结肠和直肠癌、肝癌、胰腺癌与壶腹周围癌、胆道肿瘤或小肠恶性肿瘤;所述泌尿生殖系统肿瘤为肾癌、膀胱癌、睾丸恶性肿瘤、阴茎癌、子宫颈癌、子宫内膜癌或卵巢癌;所述淋巴及血液系统肿瘤为淋巴瘤、急性白血病、慢性白血病、红细胞增多症、血小板增多症、骨髓增生异常综合症、骨髓纤维化或恶性组织细胞病。
4.如权利要求1所述的用途,其特征在于,所述肿瘤为实体瘤、淋巴瘤或白血病;优选地,所述肿瘤为肺癌、肝癌、胃癌、结肠癌、食管癌、膀胱癌、胰腺癌、脑瘤、乳腺癌、甲状腺癌、淋巴瘤、子宫颈癌或子宫体癌中的一种或几种。
7.如权利要求1-6任一项所述的用途,其特征在于,所述中药组合物的制备方法包括以下步骤:
步骤A:将荆芥、防风、羌活、独活、前胡、川芎、枳壳,蒸馏提取挥发油备用,蒸馏后的药渣、蒸馏后的川芎、枳壳水溶液备用;
步骤B:将步骤A所得蒸馏后的川芎、枳壳水溶液配制成10-40%的乙醇溶液,备用;
步骤C:将茯苓、步骤A所得蒸馏后的川芎、枳壳药渣混合,以步骤B所得乙醇溶液渗漉提取,渗漉液备用;
步骤D:将柴胡、桔梗、甘草、步骤A所得蒸馏后的荆芥、防风、羌活、独活、前胡药渣,加水煎煮,煎煮液浓缩后备用;
步骤E:将步骤C所得渗漉液、步骤D所得煎煮液混合,浓缩,加入步骤A所得挥发油,即得;
优选地,步骤B为将步骤A所得蒸馏后的川芎、枳壳水溶液配制成15-30%的乙醇溶液,备用;
进一步优选地,步骤B为将步骤A所得蒸馏后的川芎、枳壳水溶液配制成25%的乙醇溶液,备用。
8.一种中药制剂,其特征在于,所述中药制剂含有如权利要求1-7任一项所述的中药组合物;优选地,所述的中药制剂为临床上可接受的口服制剂;更优选地,所述临床上可接受的口服制剂为丸剂、胶囊剂、片剂、颗粒剂或液体口服制剂中的一种或多种;更优选地,所述口服制剂为颗粒剂;更优选地,所述颗粒剂为荆防颗粒。
9.如权利要求8所述的中药制剂,其特征在于,所述的中药制剂和至少一种药学上可接受的辅料制备成液体制剂、固体和半固体制剂、或者气体制剂;优选地,所述液体制剂是口服液、混悬液、糖浆、注射液、药酒或者酊剂,所述的固体和半固体制剂是片剂、丸剂、膏剂、丹剂、散剂、颗粒剂、栓剂、粉剂、乳剂、咀嚼剂或者胶囊剂,所述的气体制剂是气雾剂或者吸入剂。
10.如权利要求1-9任一项所述的用途,其特征在于,所述中药组合物按生药计用量为0.02g/kg·d~3.0g/kg·d;优选地,所述中药组合物按生药计用量为0.05g/kg·d~1.4g/kg·d。进一步优选地,所述中药组合物按生药计用量为0.29g/kg·d。
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