CN109381679A - 具有防治肝癌前病变及肝癌作用的中药组合物及其制备方法 - Google Patents
具有防治肝癌前病变及肝癌作用的中药组合物及其制备方法 Download PDFInfo
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Abstract
本发明属于中药制剂技术领域,具体涉及一种防治肝癌前病变及肝癌的中药组合物,并进一步公开其制备方法。本发明所述防治肝癌前病变及肝癌的药物组合物,以桑寄生、生黄芪、丹参、郁金、苦参、莪术、马鞭草、鹅去氧胆酸为原料药,利用各原料药之间君臣佐使的严格配比,充分发挥了对肝癌前病变的防治作用,以及对肝癌的治疗作用,对肝癌细胞具有明显的抑制效果。
Description
技术领域
本发明属于中药制剂技术领域,具体涉及一种具有防治肝癌前病变及肝癌作用的中药组合物,并进一步公开其制备方法。
背景技术
癌症即恶性肿瘤,是目前世界上难于治愈的疾病之一,它是100多种相关疾病的统称。人们身体内所有器官都是由细胞组成,细胞增长和分化可满足身体需要,这种有序的过程可保持人们身体健康。当身体内细胞发生突变后,它会不断地分裂,不受身体控制,这些额外的大量细胞就形成肿瘤,形成癌症。恶性肿瘤的细胞能侵犯、破坏邻近的组织和器官;而且,癌细胞可从肿瘤中穿出,进入血液或淋巴系统,这就是癌症如何从原发的部位到其它器官形成新的肿瘤,这个过程就叫癌症转移,多数癌症是根据他们起始的器官或细胞类型来命名的。
原发性肝癌(primary carcinoma of liver,肝癌)是我国常见的恶性肿瘤之一。据20世纪90年代统计,我国肝癌的年死亡率为20.37/10万,在恶性肿瘤死亡顺位中占第2位,在城市中仅次于肺癌,农村中仅次于胃癌。
对于肝癌的治疗,就目前的治疗水平而言,其诊断已不是问题,由于血清甲胎蛋白(AFP)的临床应用和各种影像学技术的进步,特别是AFP和超声显像用于肝癌高危人群的监测,使肝癌能够在无症状和体征的亚临床期做出诊断。但是肝癌的治疗却非常困难,由于现今还没有研制出如广谱抗菌素那样能普遍杀死癌细胞的药物,所以对肝癌患者的治疗,仍是先进行手术切除,然后再通过放射疗法或者化学疗法杀灭癌细胞。但这两种方法在杀灭癌细胞的同时也会杀灭正常的健康细胞,致使癌症患者的抵抗力和免疫力下降,根治性切除后易导致癌症的复发,而且治疗的价格昂贵、费用高,一般家庭难以承担。
近十年来国内外的资料表明,单纯依靠外科手段来提高肝癌的治愈率、降低手术后复发率,已处于较为困难的阶段,探讨非手术治疗是当前提高癌症疗效的一个重要途径。
发明内容
为此,本发明所要解决的技术问题在于提供一种具有防治肝癌前病变及肝癌作用的中药组合物,以解决现有技术中手术治疗肝癌治愈率较低、复发率较高的问题。
为解决上述技术问题,本发明所述的防治肝癌前病变及肝癌的中药组合物,所述中药组合物的原料药组成为:桑寄生10-15重量份、生黄芪10-15重量份、丹参5-15重量份、郁金10-15重量份、苦参5-15重量份、莪术10-15重量份、马鞭草3-8重量份、鹅去氧胆酸0.5-2重量份。
优选的,所述中药组合物的原料药组成为:桑寄生12重量份、生黄芪12重量份、丹参9重量份、郁金9重量份、苦参9重量份、莪术9重量份、马鞭草6重量份、鹅去氧胆酸1重量份。
所述中药组合物添加常规辅料,按照常规工艺制成临床上可接受的制剂。
本发明还公开了由所述的防治肝癌前病变及肝癌的中药组合物,添加常规辅料,按照常规工艺制成的临床上可接受的制剂。
所述制剂包括颗粒剂、片剂、合剂、滴丸剂、丸剂、胶囊剂。
本发明还公开了一种制备所述的具有防治肝癌前病变及肝癌作用制剂的方法,包括取选定量的桑寄生、生黄芪、丹参、郁金、苦参、莪术、马鞭草、鹅去氧胆酸混匀的步骤,并添加常规辅料,按照常规工艺制成临床上可接受的制剂。
优选的,所述的制备具有防治肝癌前病变及肝癌作用制剂的方法,包括如下步骤:
(1)取选定量的桑寄生、生黄芪、丹参、郁金、苦参、莪术、马鞭草混匀,加入水进行煎煮提取,收集煎煮液,并浓缩至清膏,备用;
(2)取所得清膏加入所述鹅去氧胆酸,并添加常规辅料,按照常规工艺制成临床上可接受的制剂。
更优的,所述的制备防治肝癌前病变及肝癌制剂的方法,包括如下步骤:
(1)取选定量的桑寄生、生黄芪、丹参、郁金、苦参、莪术、马鞭草混匀,加入占药物总量2-4重量倍量的水进行煎煮提取,收集煎煮液,并浓缩至清膏,备用;
(2)取所得清膏加入所述鹅去氧胆酸,并添加常规辅料,按照常规工艺制成临床上可接受的制剂。
更优的,所述步骤(1)中,所述煎煮步骤具体为每次加入药物总量1-2重量倍量的水进行煎煮2次。
本发明还公开了所述中药组合物用于制备具有防治肝癌前病变及肝癌作用药物的用途。
本发明所述防治肝癌前病变及肝癌的药物组合物,以桑寄生、生黄芪、丹参、郁金、苦参、莪术、马鞭草、鹅去氧胆酸为原料药,其中,
桑寄生,味苦,性平,入肝、肾经;具有补肝肾、强筋骨、祛风湿、安胎元之功效;主治腰膝酸痛、筋骨无力、头晕目眩、风湿痹痛等症;
生黄芪,味甘、微温,归肺、脾、肝、肾经;具有益气固表、敛汗固脱、托疮生肌、利水消肿之功效;用于治疗气虚乏力、内热消渴等症;
丹参,味苦、微寒,入心、肝经;具有活血散淤、消肿止血、消炎止痛、调经止痛、扩张冠状动脉、改善心肌缺血状况、降低血压、安神静心、降血糖和抗菌等功效;对症瘕积聚,胸腹刺痛,热痹疼痛,疮疡肿痛,心烦不眠;肝脾肿大,心绞痛等病症有一定的疗效;
郁金,味辛、苦,性凉,入心、肺、肝经;具有行气解郁、凉血破瘀之功效;主治失心癫狂、热病神昏等症;
苦参,味苦、性寒,入肝、肾、大肠、小肠经、心经、膀胱经;具有清热燥湿之功效;主治湿热泻痢、肠风便血、黄疸、小便不利等症;
莪术,主治气血凝滞、心腹胀痛、症瘕、宿食不消、跌打损伤作痛等症;
马鞭草,味苦、性凉,归肝、脾经;具有活血散瘀、截疟、解毒、利水消肿之功效;主治症瘕积聚、经闭痛经、疟疾、喉痹、痈肿、水肿、热淋等症;
鹅去氧胆酸(CDCA),是由胆汁提取而来,为无色针状结晶,无臭、味苦,几乎不溶于水,易溶于乙醇、冰乙酸、微溶于氯仿;鹅去氧胆酸主要作用是降低胆汁内胆固醇的饱和度,绝大多数患者服用CDCA后(当CDCA占胆汁中胆盐的70%时),脂类恢复微胶粒状态,胆固醇就处于不饱和状态,从而使结石中的胆固醇溶解、脱落。大剂量的CDCA(每日10~15mg/kg)可以抑制胆固醇的合成,并增加胆石症患者胆汁的分泌,但其中的胆盐和磷脂分泌量维持不变。中医理论认为虚证是肿瘤发生发展的重要原因和病机,因正气不足、气血虚弱、导致脏腑功能失调,因而出现气滞、血瘀、湿聚、痰结等一系列病理变化,并最终形成肿瘤。临床实践也充分证明扶正培本可以缓解症状、提高生存质量、延长生存期、降低放化疗的毒副作用等,对于某些肿瘤可有效降低复发率、提高治愈率的作用。
本发明所述防治肝癌前病变及肝癌的药物组合物,利用各原料药之间君臣佐使的严格配比,具有扶正培本、益气补肾、利湿解毒和活血散结之功效。药理实验显示,本发明所述药物组合物可有效抑制肿瘤增长及促进肿瘤细胞凋亡,充分发挥了对肝癌前病变的防治作用,对肝癌细胞具有明显的抑制效果,可有效用于肝癌的治疗作用。同时,所选用的原料来源广泛易得,尤其是选用的鹅去氧胆酸可直接进行化学合成,进一步降低到了工艺成本。
附图说明
为了使本发明的内容更容易被清楚的理解,下面根据本发明的具体实施例并结合附图,对本发明作进一步详细的说明,其中,
图1为HepG2细胞用药前后的细胞形态对比图;
图2为HepG2肝癌细胞系的成瘤及抑瘤检测结果。
具体实施方式
实施例1颗粒剂
本实施例所述防治肝癌前病变及肝癌的颗粒剂的原料药组成为:桑寄生10g、生黄芪15g;丹参5g;郁金15g;苦参5g;莪术15g;马鞭草3g;鹅去氧胆酸2g。
本实施例所述防治肝癌前病变及肝癌的颗粒剂的制备方法,包括如下步骤:
(1)取选定量的桑寄生、生黄芪、丹参、郁金、苦参、莪术、马鞭草混匀,加入占药物总量2重量倍量的水进行煎煮提取,收集煎煮液,并浓缩至清膏,备用;
(2)取所得清膏加入所述鹅去氧胆酸,并添加常规辅料,按照常规工艺制成临床上可接受的颗粒剂。
实施例2片剂
本实施例所述防治肝癌前病变及肝癌的片剂的原料药组成为:桑寄生15g、生黄芪10g;丹参15g;郁金10g;苦参15g;莪术10g;马鞭草8g;鹅去氧胆酸0.5g。
本实施例所述防治肝癌前病变及肝癌的片剂的制备方法,包括如下步骤:
(1)取选定量的桑寄生、生黄芪、丹参、郁金、苦参、莪术、马鞭草混匀,加入占药物总量4重量倍量的水进行煎煮提取,收集煎煮液,并浓缩至清膏,备用;
(2)取所得清膏加入所述鹅去氧胆酸,并添加常规辅料,按照常规工艺制成临床上可接受的片剂。
实施例3颗粒剂
本实施例所述防治肝癌前病变及肝癌的颗粒剂的原料药组成为:桑寄生12g、生黄芪12g、丹参9g、郁金9g、苦参9g、莪术9g、马鞭草6g、鹅去氧胆酸1g。
本实施例所述防治肝癌前病变及肝癌的颗粒剂的制备方法,包括如下步骤:
(1)取选定量的桑寄生、生黄芪、丹参、郁金、苦参、莪术、马鞭草混匀,加入占药物总量3重量倍量的水进行煎煮提取,收集煎煮液,并浓缩至清膏,备用;
(2)取所得清膏加入所述鹅去氧胆酸,并添加常规辅料,按照常规工艺制成临床上可接受的颗粒剂。
实施例4胶囊剂
本实施例所述防治肝癌前病变及肝癌的胶囊剂的原料药组成为:桑寄生11g、生黄芪13g、丹参7g、郁金12g、苦参7g、莪术12g、马鞭草5g、鹅去氧胆酸1.5g。
本实施例所述防治肝癌前病变及肝癌的胶囊剂的制备方法,包括如下步骤:
(1)取选定量的桑寄生、生黄芪、丹参、郁金、苦参、莪术、马鞭草混匀,加入占药物总量3重量倍量的水进行煎煮提取,收集煎煮液,并浓缩至清膏,备用;
(2)取所得清膏加入所述鹅去氧胆酸,并添加常规辅料,按照常规工艺制成临床上可接受的胶囊剂。
实施例5丸剂
本实施例所述防治肝癌前病变及肝癌的丸剂的原料药组成为:桑寄生13g、生黄芪11g、丹参12g、郁金9.5g、苦参12g、莪术9.5g、马鞭草7g、鹅去氧胆酸0.8g。
本实施例所述防治肝癌前病变及肝癌的丸剂的制备方法,包括如下步骤:
(1)取选定量的桑寄生、生黄芪、丹参、郁金、苦参、莪术、马鞭草混匀,加入占药物总量3重量倍量的水进行煎煮提取,收集煎煮液,并浓缩至清膏,备用;
(2)取所得清膏加入所述鹅去氧胆酸,并添加常规辅料,按照常规工艺制成临床上可接受的丸剂。
实验例
一、实验试剂与仪器
1.实验主要试剂
2.实验主要仪器
仪器名称 | 仪器型号 | 厂家 |
二氧化碳培养箱 | MCO-15AC型 | SANYO |
倒置显微镜 | XDS-2B型 | 重庆光电 |
超净台 | SW-CJ-1D型 | 江苏通净 |
离心机 | Centrifuge 5415D | Eppendorf |
酶标仪 | MultiSkan3 | Thermo |
层流柜 | 苏杭 |
二、实验方法
1、供试药物
取桑寄生12g、生黄芪12g、丹参9g、郁金9g、苦参9g、莪术9g、马鞭草6g、鹅去氧胆酸1g,混匀并加入占药物总量3重量倍量的水进行煎煮提取,收集煎煮液。
2、细胞培养及收集
2.1细胞复苏
42℃水浴在1min内快速溶解冻存细胞,放置于T-25培养瓶培养。
2.2细胞日常维护
(1)将细胞原生长培养基弃掉,用无血清培养基清洗2次;
(2)加入0.25%胰酶,1ml/T-25,以盖住细胞容器底部为准,37℃孵育1-3min;
(3)加入5ml生长培养基,重悬细胞,对细胞进行计数,1000rpm离心5min;
(4)弃上清,细胞沉淀可进行冻存、用于药物筛选或收集细胞。
各种肝癌细胞系对应生长培养基如下表1。
表1各种肝癌细胞系对应生长培养基
英文名称 | 中文名称 | 生长特点 | 培养条件 |
L-02 | 人正常肝细胞 | 贴壁生长 | 含10%FBS的1640 |
HepG2 | 人肝癌细胞 | 贴壁生长 | 含10%FBS的1640 |
MHCC97-H | 高转移人肝癌细胞 | 贴壁生长 | 含10%FBS的DMEM |
Hcclm3 | 人高转移肝癌细胞 | 贴壁生长 | 含10%FBS的1640 |
SMMC-7721 | 人肝癌细胞 | 贴壁生长 | 含10%FBS的1640 |
HCCC-9810 | 人胆管细胞型肝癌细胞 | 贴壁生长 | 含10%FBS的1640 |
Hep B1.2 | 人肝癌细胞 | 贴壁生长 | 含10%FBS的MEM-EBSS |
Hep3b | 人肝癌细胞 | 贴壁生长 | 含10%FBS的MEM-EBSS |
HuH-6 | 人肝母细胞瘤细胞; | 贴壁生长 | 含10%FBS的1640 |
HuH-7 | 人肝癌细胞 | 贴壁生长 | 含10%FBS的1640 |
SK-HEP-1 | 人肝腹水腺癌细胞 | 贴壁生长 | 含10%FBS的1640 |
HB611 | 人肝癌细胞 | 贴壁生长 | 含10%FBS的DMEM |
BEL-7402 | 人肝癌细胞 | 贴壁生长 | 含10%FBS的1640 |
BEL-7404 | 人肝癌细胞 | 贴壁生长 | 含10%FBS的1640 |
BEL-7405 | 人肝癌细胞 | 贴壁生长 | 含10%FBS的1640 |
PLC/PRF/5 | 人肝癌细胞 | 贴壁生长 | 含10%FBS的MEM-EBSS |
3、药物对正常肝细胞的无毒浓度筛选
(1)将L02细胞原培养液弃掉,用0.25%胰酶37℃消化细胞2min后,加入生长培养基重悬细胞,调整细胞密度为5×104个/ml;
(2)加入96孔细胞培养板内,100ul/孔,继续培养24h;
(3)更换无血清培养,同步化24h;
(4)用不同浓度供试药物(浓度为10%、2%、0.4%、0.08%、0.016%)作用细胞48h;
(5)48h后弃原有培养基,加入100ul培养基和10ul MTT,37℃反应4h;
(6)弃培养基,加入100ul 10%SDS溶解细胞,570nm下读取吸光值,并计算细胞抑制率,筛选出A对正常肝细胞的无毒浓度;
细胞抑制率=(对照OD值-实验组OD值)/对照OD值×100%。
4、药物对正常细胞无毒浓度下对肝癌细胞的生长抑制效果检测
(1)将所有肝癌细胞原培养液弃掉,用0.25%胰酶37℃消化细胞2min后,加入生长培养基重悬细胞,调整细胞密度为5×106个/ml;
(2)加入96孔细胞培养板内,100ul/孔,继续培养24h;
(3)更换无血清培养,同步化24h;
(4)用方法上述3项中方法筛选获得的无毒浓度作用肝癌细胞48h;
(5)弃原有培养基,加入100ul培养基和10ul MTT,37℃反应4h;
(6)弃培养基,加入100ul 10%SDS溶解细胞,570nm下读取吸光值,并计算细胞抑制率,观察对各种肝癌细胞的生长抑制率,每次3孔重复,至少重复3次;
细胞抑制率=(对照OD值-实验组OD值)/对照OD值×100%。
5、细胞收集、接种
(1)将HepG2细胞原生长培养基弃掉,用无血清培养基清洗2次;
(2)加入0.25%胰酶,1ml/T-25,以盖住细胞容器底部为准,37℃孵育1-3min;
(3)加入5ml生长培养基,重悬细胞,对细胞进行计数,1000rpm离心5min;
(4)弃上清,用10mM PBS洗涤细胞两次,每次1000rpm离心5min;
(5)用10mM PBS调整细胞密度为5×107个/ml;
(6)按5×106个/0.1ml/只的量接种于B/C裸鼠腋下;
(7)接种7-10天后,肉眼可见的肿瘤时,随机分为两组:中药组,按1.0ml/只/天灌胃A药;对照组,按1.0ml/只/天灌胃10mM无菌PBS;
(8)经18天的持续灌胃后,对比观察两组腋下肿瘤组织肿瘤、大小,确认A药对肝癌细胞的抑制作用。
三、实验结果
1、正常肝细胞无毒浓度摸索结果
按照上述方法检测正常肝细胞无毒浓度结果,见下表2。
表2正常肝细胞无毒浓度摸索结果
2、肝癌细胞抑制效果检测结果
按照上述方法检测肝癌细胞抑制效果,检测结果见下表3。
表3肝癌细胞抑制效果检测结果
同时对HepG2细胞在中药作用前后的细胞形态进行拍照,观察中药对细胞生长的抑制情况,细胞形态结果见图1。
3、裸鼠成瘤及抑瘤结果
选择HepG2肝癌细胞系进行成瘤、抑瘤效果检测,HepG2细胞的成瘤情况及用药情况检测结果见图2。可见,中药用药组肿瘤明显被抑制。
显然,上述实施例仅仅是为清楚地说明所作的举例,而并非对实施方式的限定。对于所属领域的普通技术人员来说,在上述说明的基础上还可以做出其它不同形式的变化或变动。这里无需也无法对所有的实施方式予以穷举。而由此所引伸出的显而易见的变化或变动仍处于本发明创造的保护范围之中。
Claims (10)
1.一种具有防治肝癌前病变及肝癌作用的中药组合物,其特征在于,所述中药组合物的原料药组成为:桑寄生10-15重量份、生黄芪10-15重量份、丹参5-15重量份、郁金10-15重量份、苦参5-15重量份、莪术10-15重量份、马鞭草3-8重量份、鹅去氧胆酸0.5-2重量份。
2.根据权利要求1所述的具有防治肝癌前病变及肝癌作用的中药组合物,其特征在于,所述中药组合物的原料药组成为:桑寄生12重量份、生黄芪12重量份、丹参9重量份、郁金9重量份、苦参9重量份、莪术9重量份、马鞭草6重量份、鹅去氧胆酸1重量份。
3.根据权利要求1或2所述的具有防治肝癌前病变及肝癌作用的中药组合物,其特征在于,所述中药组合物添加常规辅料,按照常规工艺制成临床上可接受的制剂。
4.由权利要求1-3任一项所述的具有防治肝癌前病变及肝癌作用的中药组合物,添加常规辅料,按照常规工艺制成的临床上可接受的制剂。
5.根据权利要求4所述的制剂,其特征在于,所述制剂包括颗粒剂、片剂、合剂、滴丸剂、丸剂、胶囊剂。
6.一种制备权利要求4或5所述的具有防治肝癌前病变及肝癌作用制剂的方法,其特征在于,包括取选定量的桑寄生、生黄芪、丹参、郁金、苦参、莪术、马鞭草、鹅去氧胆酸混匀的步骤,并添加常规辅料,按照常规工艺制成临床上可接受的制剂。
7.根据权利要求6所述的制备具有防治肝癌前病变及肝癌作用制剂的方法,其特征在于,包括如下步骤:
(1)取选定量的桑寄生、生黄芪、丹参、郁金、苦参、莪术、马鞭草混匀,加入水进行煎煮提取,收集煎煮液,并浓缩至清膏,备用;
(2)取所得清膏添加所述鹅去氧胆酸混匀,并添加常规辅料,按照常规工艺制成临床上可接受的制剂。
8.根据权利要求6或7所述的制备具有防治肝癌前病变及肝癌作用制剂的方法,其特征在于,包括如下步骤:
(1)取选定量的桑寄生、生黄芪、丹参、郁金、苦参、莪术、马鞭草混匀,加入占药物总量2-4重量倍量的水进行煎煮提取,收集煎煮液,并浓缩至清膏,备用;
(2)取所得清膏加入所述鹅去氧胆酸混匀,并添加常规辅料,按照常规工艺制成临床上可接受的制剂。
9.根据权利要求8所述的制备具有防治肝癌前病变及肝癌作用制剂的方法,其特征在于,所述步骤(1)中,所述煎煮步骤具体为每次加入药物总量1-2重量倍量的水进行煎煮2次。
10.权利要求1-3任一项所述中药组合物用于制备具有防治肝癌前病变及肝癌作用药物的用途。
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CN1985981A (zh) * | 2006-12-20 | 2007-06-27 | 北京润德康医药技术有限公司 | 一种用于各种癌症及其恢复期等疾病的中药组合物 |
CN104707097A (zh) * | 2013-12-17 | 2015-06-17 | 首都医科大学 | 药物组合物槲芪散及其在制备用于阻断肝癌前病变、治疗肝癌或病毒性肝炎的药物中的应用 |
CN105348354A (zh) * | 2014-12-03 | 2016-02-24 | 四川百利药业有限责任公司 | 一类鹅去氧胆酸类化合物及其制备方法和用途 |
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CN1985981A (zh) * | 2006-12-20 | 2007-06-27 | 北京润德康医药技术有限公司 | 一种用于各种癌症及其恢复期等疾病的中药组合物 |
CN104707097A (zh) * | 2013-12-17 | 2015-06-17 | 首都医科大学 | 药物组合物槲芪散及其在制备用于阻断肝癌前病变、治疗肝癌或病毒性肝炎的药物中的应用 |
CN105348354A (zh) * | 2014-12-03 | 2016-02-24 | 四川百利药业有限责任公司 | 一类鹅去氧胆酸类化合物及其制备方法和用途 |
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