CN105288453A - 预防癌症、治疗癌症及增强其他抗癌药物效果的组合物 - Google Patents
预防癌症、治疗癌症及增强其他抗癌药物效果的组合物 Download PDFInfo
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Abstract
本发明涉及一种来自中草药、药用植物及其萃取物的组合物,尤其是一种用于预防癌症、治疗癌症及增强其他抗癌药物效果的组合物。具体而言,本发明的组合物来自人类长期使用的各种中草药或药用植物。本发明的组合物通过特殊技术制得且于细胞毒杀试验中显示出对恶性黑色素瘤细胞、前列腺癌细胞、大肠癌细胞、肺癌细胞、胃癌细胞、乳癌细胞及肝癌细胞均具有良好的毒杀癌细胞活性。此外还可明显地增强其他抗癌药物效果。而癌症对医药团体提出的特殊问题,也为本发明所专注。
Description
技术领域
本发明涉及一种来自中草药、药用植物及其萃取物的组合物,尤其是一种用于预防癌症、治疗癌症及增强其他抗癌药物效果的组合物。更具体的,本发明的组合物来自人类长期使用的各种中草药或药用植物。本发明的组合物通过特殊技术制备且于细胞毒杀试验中显示出对恶性黑色素瘤细胞、前列腺癌细胞、大肠癌细胞、肺癌细胞、胃癌细胞、乳癌细胞及肝癌细胞均具有良好的毒杀癌细胞活性。并且可明显地增强其他抗癌药物效果。而癌症对医药团体提出的特殊问题,也为本发明所专注。
现代医药科学不断地寻找新颖且更有效的药剂以预防及治疗癌症。人类每年花费数十亿甚至数百亿美元。每年由制药公司花费巨资于鉴定、特性化及制造新抗癌药物以对抗不断出现的具抗药性的癌细胞,早已成为严重的问题。预防癌症的有效方法仍是人们主要关注的。
本发明涉及一种组合物,一种用于预防癌症、治疗癌症及增强其他抗癌药物效果的组合物。本发明公开的组合物中各物质重量比大约为夏枯草(PRUNELLAE SPICA): 半枝莲(SCUTELLARIAE BARBATAE
HERBA): 牛樟芝(ANTRODIA CAMPHORATA):姜黄(CURCUMAE RHIZOMA): 牡丹皮(PAEONIAE SUFFRUTICOSAE
CORTEX): 败酱草(PATRINIAE HERBA): 小金英(IXERIS HERBA): 桑黄(PHELLINUS
LINTEUS)=(0.5~5.0):(0.5~5.0):(0.5~5.0):(0.5~5.0):(0.5~5.0):(0.5~5.0):(0.5~5.0):(0.5~5.0)。
癌症为人类十大死因之首,其特征为恶性组织的不正常肿块,起因于过度的细胞分裂。癌症细胞不具有正常细胞生长的限制,会不正常地侵入其他细胞。此处所指的癌症包括所有种类的细胞不当增生或致癌过程或转移。一般所谓的癌症包括脑癌、头部与颈部的癌症、眼癌(例如视网膜母细胞瘤)、鼻咽癌、口腔癌、食道癌、肺癌、支气管癌、间皮( Mesothelioma) 、甲状腺癌、乳癌、胃癌、肝癌、胰脏癌、胆管癌、肾脏癌、肾上腺皮质癌、嗜铬细胞瘤、卵巢癌、子宫内膜癌、子宫颈癌、大肠直肠癌、膀胱癌、前列腺癌、阴茎癌、睪丸癌、阴道癌、血癌、肌肉癌(例如恶性肉瘤)、骨癌、关节癌、淋巴瘤、皮肤癌(例如恶性黑色素瘤)、神经癌(例如神经胶质瘤)、转移癌与恶性肉瘤(Sarcoma)。
癌症治疗的种类包括化学治疗、手术治疗、放射线治疗、贺尔蒙疗法、生物制剂疗法、标靶治疗以及这些治疗的结合疗法。化学治疗一般使用一个或多个抑制癌细胞生长的化合物。
虽然目前已经研制了许多癌症化疗药物,但由于副作用、有限的疗效及抗药性等原因仍然需要更有效的化疗药物。此外更有人选择以较为温和的中医疗法进行癌症治疗。中药是民众普遍认为对身体较温和的治疗药物,并具有较高的市场接受度。因此,若能研制出一种药学组合物,其经实验证明确实可毒杀癌细胞,必可对于癌症治疗具有相当的帮助。
本发明的组合物,可经口或经鼻或经皮肤或经注射或经肛门或经生殖器而给药或通过吸入喷雾或植入贮存器的方式或以手术等侵入性方式施用于内脏组织的患部而给药或以针炙穴位经络给药。当本发明的组合物经口给予时,可选自以下群中的产品形式:糖果、甜点、凝胶、营养补充品、口香糖、医药、婴儿营养品、饮料、优格、牛奶、再水合溶液或水溶液。
草药及民间药的使用在中国已有几千年历史。此等草药可治疗无数疾病,从关节炎至病毒感染等以前被现代西方医学视为无效且危险的疾病。于19世纪当中,许多含草药的家庭医疗药已申请专利及贩卖。现代药已代替那些家庭医疗药,但许多现代药含有草药成分。例如,1776年,英国植物学家及医生William Withering发现一种由老农妇制造的草药,能有效治疗水肿,即排出组织中过剩的水,此病由心脏不够强无法打出血液所引起。其发现药中的一种成分,由指顶花植物的叶制成,能强化心脏打出能力。由指顶花植物制成的药现称为毛地黄。此外 Monroe E. Wall及 Mansukh C. Wani于1967年于美国国家癌症研究所,从太平洋紫杉(短叶豆杉)树皮中分离出化疗药物紫杉醇 (taxol)。
民间药对于西方而言为相对现代的用词,而已通过各种草药照护及治疗疾病。近年来,民间药已于西方科学医药界获得更多关注。
背景技术
夏枯草(PRUNELLAE SPICA),制备自滁州夏枯草(Prunella vulgaris)或日本夏枯草(Prunella vulgaris subsp.
asiatica)(亦称为Prunella vulgaris vra.
lilachina)的干燥花穗或全草。两植物皆属唇形科。此草药具苦味且无毒。典型的平均治疗剂量为每人每日4至110克。临床上可治疗高血压、急性结膜炎、急性黄疸性肝炎、颈部淋巴炎、淋巴结核、单纯性甲状腺肿、扁桃腺炎与咽喉炎。亦可配合其他中草药治疗眼珠疼痛、流泪惧光、腹痛、疮疡与子宫出血。夏枯草可抑制痢疾杆菌、伤寒杆菌、霍乱弧菌、大肠杆菌、变形杆菌、绿脓杆菌、葡萄球菌与链球菌。Hozumi等公开了滁州夏枯草的花穗作为抗疱疹病毒剂可用于治疗疱疹病毒感染。参见T. Hozumi等人,美国专利第5,411,733号,1995年5月2日公告。蔡修贤等人公开了夏枯草的水萃取物于2.5 mg/ml浓度时于试管试验第三天可抑制100 %艾滋病毒活性,而第四天仍可抑制100 %艾滋病毒活性。参见蔡修贤等人,美国专利第5837257号,1998年11月17日公告。此外夏枯草亦可配合其他中草药治疗癌症:夏枯草与蒲公英并用可以治疗早期乳癌。参见张宪昌药草(一) 渡假出版社有限公司 台北,中国台湾,第15页(1997)。李健勇公开了一种由党参、黄芪、薏苡仁、山佩兰、橘皮、爵床、夏枯草、凉粉草、栀子、连翘、赤芍、大黄、蟅虫 、白花蛇舌草、扛板归、山豆根、木槿皮、天南星、半边莲、白蔹、漏芦、石苇、乳香、没药、蚤休、败酱、白芷、射干、蒲公英、螺厣草、皂荚、独活、麻黄及升麻等组成的中药复方可用于治疗淋巴结关系型癌症。参见李健勇,台湾专利第I260226号,2006年08月21日公告。Y. Maimon 公开了一种由黄芪、茯苓、白术、枸杞、女贞子、白芍、北沙参、陈皮、麦冬、鸡血藤、白花蛇舌草、半枝莲及夏枯草所组成的组合物可用于减少与化疗相关的副作用。参见Y. Maimon,美国专利第8,574,636号,2013年11月5日公告。
半枝莲(SCUTELLARIAE BARBATAE HERBA),制备自半枝莲(Scutellaria barbata)、半枝莲(Scutellaria rivularis)或狗牙半枝莲(Scutellaria dependens),其属于唇形科。此草药具苦味且不宜于贫血者服用。怀孕妇人应避免服此药草。典型治疗剂量为每日4至300克。临床上可治跌打损伤、惊风、肺炎、喉痛、腹痛 与肠炎。蔡修贤等人公开了半枝莲的水萃取物以2.5 mg/ml的浓度于试管试验第三天可抑制92%艾滋病毒活性,而第四天可抑制94%艾滋病毒活性。参见蔡修贤等人,美国专利第5837257号,1998年11月17日公告。I. Cohen 公开了半枝莲萃取物(未并用任何其他中草药)可治疗晚期乳癌与转移性乳癌。参见I. Cohen,美国专利第 7700136号,2010年4月20日公告。此外半枝莲亦可配合其他中草药治疗癌症:A. S. Sun 公开了白花蛇舌草、半枝莲、绿豆与香菇可改善恶性肿瘤。参见 A. S. Sun,美国专利第5437866号,1995年8月1日公告。J. Dao等人公开了灵芝、丹蔘与半枝莲的乙酸乙酯萃取物可治疗癌症。参见 J. Dao 等人,美国专利第8173177号,2012年5月8日公告。Y. Maimon公开了一种由夏枯草、黄芪、茯苓、白术、枸杞、女贞子、白芍、北沙参、陈皮、麦冬、鸡血藤、白花蛇舌草及半枝莲所组成的组合物可用于减少与化疗相关的副作用。参见Y. Maimon,美国专利第8,574,636号,2013年11月5日公告。
牛樟芝( ANTRODIA CAMPHORATA ),属于非褶菌目( Aphyllophorales )多孔菌科( Polyporaceae )的多年生蕈菌类。牛樟芝制备于牛樟芝的子实体、菌丝体或二者的混合物。由牛樟芝萃取的各种化合物有保护肝脏、抗氧化、抗炎与抗癌等各种不同的作用。G. Hsiao等人指出,牛樟芝萃取物能有效预防四氯化碳所诱导的肝毒性。参见G. Hsiao et al.,
Antioxidative and hepatoprotective effects of Antrodia camphorate extract.J.
Agric. Food Chem., 2003, 51: 3302–3308。W. C. Lin等人指出,由牛樟芝发酵的菌丝体能减缓四氯化碳所诱导大鼠肝纤维化的进展。参见 W. C. Lin et al. Filtrate
of fermented mycelia from Antrodia camphorata reduces liver fibrosis induced by
carbon tetrachloride in rats World J Gastroenterol 2006 April ;
12(15):2369-2374。S. Y. Liu等人公开了牛樟芝的环己烯酮化合物能有效地抑制HBsAg(B型肝炎表面抗原)与 HBeAg(B型肝炎e抗原)以达成抑制HBV(B型肝炎病毒)的目的。参见S. Y.Liu et al.,美国专利第7411003号,2008年8月12日公告。S. Y.Liu等人公开了牛樟芝的环己烯酮化合物能减少肝脏损伤和自由基引起的氧化压力,增强抗氧化能力,并能达到保护肝脏的目的。参见S. Y.Liu et al.,美国专利第7456225号,2008年11月25日公告。C. H. HUANG等人指出,高剂量(0.1 g/kg BW/day)牛樟芝对被喂酒精的老鼠不只高胆固醇起作用,也可减少肝脏的脂质。参见CH Huang et al. , Fruiting
body of Niuchangchih (Antrodia camphorata) protects livers against chronic
alcohol consumption damage. J Agric Food Chem. 2010 Mar 24; 58(6):3859-66。T. Y. SONG等人指出,牛樟芝深层培养于不同溶剂萃取物中发酵滤液的干物质(DMF)具有最强的抗氧化活性。参见T.Y. SONG et al.,
Protective Effects of Fermented Filtrate from Antrodia camphorata in Submerged
Culture against CCl4-Induced Hepatic Toxicity in Rats. J. Agric. Food Chem.
2003, 51, 1571-1577。Chuian-Fu KEN等人指出,重组的牛樟芝-过氧化氢酶(从牛樟芝所得的过氧化氢酶)可以起酵素作用以解毒·OH基。参见Chuian-Fu KEN et al.Biochemical
characterization of a catalase from Antrodia camphorata: Expression in
Escherichia coli and enzyme properties Botanical Studies (2008) 49: 119-125。Y. k. Rao等人指出,牛樟芝的氯仿和甲醇萃取物的抗炎作用,是通过抑制巨噬细胞介导的炎症介质实现的。参见Y.k. Rao et al.,
Evaluation of the anti-inflammatory and anti-proliferation tumoral cells
activities of Antrodia camphorata, Cordyceps sinensis, and Cinnamomum
osmophloeum bark extracts. Journal of Ethnopharmacology, 2007 Oct 114(1), 78-85。S. Y.Liu等人公开了牛樟芝的环己烯酮化合物能通过促进在人体内磷酸肌酸激酶和血氨的代谢,促进他们的恢复,进一步达到延缓生理性疲劳的功能。参见S. Y.Liu et al., 美国专利第7468392号, 2008年12月23日公告。S. Y.Liu等人公开了牛樟芝的环己烯酮化合物能用以治疗自身免疫性疾病。参见S. S. Y.Liu et al., 美国专利第7501454号, 2009年3月10日公告。J. J. Liu等人指出,一种由牛樟芝菌丝体所得的独特多醣体成份于体外与体内二模式均有明显的抗肿瘤效果。参见J. J. Liu et al.,
Antitumor effects of the partially purified polysaccharides from Antrodia
camphorata and the mechanism of its action. Toxicol.
Appl. Pharmacol, 2004, 201: 186–193。C. Y. Lee公开了由白术、当归、韩信草、火炭母草、艾纳香、蒴藋、马兰、绿豆、甘蔗、大青根、半边莲、杏仁、牛樟芝、前胡、假茼蒿、茵陈蒿、牛樟芝(Nuzhenzi) 、郁金、枳实与半夏所组成的药品混合物可用于治疗肝癌与胰脏癌。参见C. Y. Lee. 美国专利第6,998,141, 号,2006年2月14日公告。Y. C. Leut等人指出,牛樟芝二甲基氧化硫萃取物对人类骨肉瘤细胞MG 63的作用,牛樟芝浓度为25~50微克/毫升时并不影响癌细胞存活率,而牛樟芝浓度为100~200微克/毫升时能降低癌细胞存活率与诱导癌细胞自杀,因此可以以浓度相关的方式降低癌细胞存活率与诱导癌细胞自杀。参见Y.C. Luet al. Effects of
Antrodia camphorata on viability, apoptosis, [Ca2+], and MAPKs phosphorylation
in MG63 human osteosarcoma cells. Drug Dev. Res., 2007, 68: 71–78。C.Y. Chang等人指出,牛樟芝乙醇萃取物与抗肿瘤药物并用时对人类肝癌细胞(体外)和植入肿瘤裸鼠异种移植细胞(体内)显示出辅助抗细胞增殖的作用。参见C.Y.Chang et al.,The adjuvant effects of Antrodia camphorate extracts
combined with anti-tumor agents on multidrug resistant human hepatoma cells. J.
Ethnopharmacol. 2008, 118: 387–395。S. Y. Liu等人公开了由牛樟芝制得的新颖化合物与用途尤其是Antroquinonol B and
Antroquinonol C,能有效地抑制癌细胞的生长。参见 S. Y.Liu et al. , 美国专利第 7,385,088号,2008年6月10日公告。S. L. Li等人指出,牛樟芝与甘草与大豆共同发酵的乙醇萃取物能增强牛樟芝对于人类肝癌细胞的抗肿瘤作用。参见S. L. Li et al., The
Augmented Anti-Tumor Effects of Antrodia camphorata Co-Fermented with Chinese
Medicinal Herb in Human Hepatoma Cells, The American Journal of Chinese
Medicine, 2009, 37, 4, 771–783。C. C. Huang等人指出,牛樟芝的二甲基氧化硫萃取物作用于人类口腔癌细胞的功效,于牛樟芝浓度为25微克/毫升时能显著增加癌细胞存活率,而于牛樟芝浓度为100-200微克/毫升时则降低癌细胞存活率,因此可以以浓度相关的方式降低癌细胞存活率。参见 C. C. Huang et al. ,
Effects of Antrodia camphorata Extracts on the Viability, Apoptosis, [Ca2+]i,
and MAPKs Phosphorylation of OC2 Human Oral Cancer Cells, Chinese Journal of
Physiology, 2009 ; 52(3): 128-135。赖基铭等人公开了乐瓦司他汀(lovastatin)(二羟甲戊酸径(mevalonate pathway)抑制剂)可显著增强牛樟芝( Antrodia camphorata )也称为“ Taiwanofungus camphoratus
”萃取物在抑制癌症细胞生长上的功效。参见赖基铭等人,台湾专利第I311912号,2009年07月21日公告。T. I. Chen等人指出,通过90天的 Sprague- Dawlly rats亚慢性毒性试验,可确认在剂量大于每天每公斤体重3公克的标准下,牛樟芝于无明显副作用。参见T. I. Chen et al., A
90-day subchronic toxicological assessment of Antrodia cinnamomea in
Sprague-Dawley rats. Food and Chemical Toxicology 2011 Feb;49(2):429-33。
姜黄(CURCUMAE RHIZOMA),制备自姜黄、郁金、黄根姜黄或莪术的干燥根茎,这些植物皆属姜科,其功能为活血、化瘀与止痛。典型的平均治疗剂量为每人每日3至12克。此外姜黄亦可与其他草药或药物并用以保护肝脏或治疗癌症。N. KAORU等人公开了一种以姜黄与鲨烯或三十碳烷的混合物为主要成分的新型含姜黄组成物,具有显著的肝功能保护及改善效果。参见N. KAORU等人,台湾专利第I277422 号,2007年04月01日 公告。C. Y. Lee公开了由白术、当归、韩信草、火炭母草、艾纳香、蒴藋、马兰、绿豆、甘蔗、大青根、半边莲、杏仁、牛樟芝、前胡、假茼蒿、茵陈蒿、郁金(Yujin)、枳实与半夏所组成的药品混合物可用于肝癌与胰脏癌。参见C. Y. Lee.美国专利第6998141号,2006年2月14日公告。T. Newmark等人公开了一种由罗勒、姜、姜黄、黄芩、迷迭香、绿茶、虎杖、 中国黄连与伏牛花的水醇萃取物所组成的组合物,可用于治疗前列腺癌。参见T. Newmark等人,美国专利第7,470,440号,2008年11月30日公告。T. Newmark等人公开了一种由罗勒、姜、姜黄、黄芩、迷迭香、绿茶、虎杖、中国黄连与伏牛花的水醇萃取物所组成的组合物,可用于治疗胶质母细胞瘤。参见 T. Newmark等人,美国专利第7,622,142号,2009年11月24日公告。T. Newmark,等人公开了一种由罗勒、姜、姜黄、黄芩、迷迭香、绿茶、虎杖、中国黄连与伏牛花的水醇萃取物所组成的组合物,可用于治疗前列腺癌。参见 T. Newmark等人,美国专利第7,470,440号,2008年11月30日公告。姜黄素为姜黄的主成份,被用为预防癌症与抗氧化的保健食品。李政育等人公开了一种由牛黄、郁金、水牛角、黄芩、黄连、雄黄、生栀子、朱砂、龙脑、麝香、真珠与金箔所组成的组合物,可用于预防脑缺血后再灌注病变。参见李政育等人,台湾专利第I353846号, 2011年12月11日公告。
牡丹皮(PAEONIAE SUFFRUTICOSAE CORTEX),制备自牡丹(Paeonia suffruticosa)、矮牡丹(Paeonia suffruticosa var.
spontanea)、紫斑牡丹(Paeonia
papaveracea)、川牡丹(Paeonia szechuanica)、黄牡丹(Paeonia lutea)、野牡丹(Paeonia delavayi)、保氏牡丹(Paeonia potanini)、茂纹牡丹(Paeonia thalictrumifolia)与云南牡丹(Paeonia yunnanonsis)的干燥根皮。此九种植物均为毛茸科。临床上用以治疗发斑、惊痫、吐血、屻血、便血、经闭、肿瘤、疮疡、细菌感染或月经过多。孕妇、腹泻与多汗者均不宜用。典型的治疗剂量为每日7至12克。谢秀梅等人公开了一种由黄莲、升麻、当归、生地黄与牡丹皮所组成的组合物,可以用于抑制肺癌干细胞生长。参见谢秀梅等人,台湾专利第I413525号,2013年11月1日公告。
败酱草(PATRINIA HERBA),制备自黄花败酱草(Patrinia scabiosaefolia)或白花败酱草(Patrinia villosa)干燥全草。典型的治疗剂量为每日10至20克。临床上可治疗感染性疾病:流行性感冒、腮腺炎、结膜炎、扁桃腺炎、急性胰脏炎、肺炎、肝脓疡、肠胃炎、吐血、流鼻血与神经衰弱。李健勇公开了一种由党参、黄芪、薏苡仁、山佩兰、橘皮、爵床、夏枯草、凉粉草、栀子、连翘、赤芍、大黄、蟅虫 、白花蛇舌草、扛板归、山豆根、木槿皮、天南星、半边莲、白蔹、漏芦、石苇、乳香、没药、蚤休、败酱草、白芷、射干、蒲公英、螺厣草、皂荚、独活、麻黄及升麻等组成的中药复方可用于治疗淋巴结关系型癌症。参见李健勇,台湾专利第I260226号,2006年08月21日公告。
小金英(IXERIS HERBA),制备自小金英(Ixeris chinensis)干燥全草。典型的治疗剂量为每日10至40克。临床上可治疗肺炎、咽喉痛、白喉、肠炎、膀胱炎、痔疮与尿路结石。此外小金英可与其他中草药并用,作为女性成长养生品。参见蔡宗义,台湾专利第M376295号,2010年03月21日公告。
桑黄(PHELLINUS LINTEUS),制备自桑黄子实体、菌丝体或二者的混合物。桑黄属于刺革菌科的蕈菌类。在日本、韩国与中国桑黄已广泛地被使用。桑黄对于各种疾病有效,像改善血液循环、促进解毒、保护人体肝脏、防治过敏、治疗糖尿病、治愈口腔溃疡与减轻胃肠障碍或淋巴疾病。桑黄萃取物被报告拥有免疫调节与抗癌作用。桑黄为抗氧化剂与肝脏保护剂的优良中草药源。参见 T. Zhu 等人,A Medicinal Mushroom:
Phellinus linteus , Current Medicinal Chemistry 2008,15: 1330~1335。
发明内容
本发明涉及一种组合物,尤其涉及一种用于预防癌症、治疗癌症及增强其他抗癌药物效果的组合物。本发明公开的组合物中各物质的重量比大约为夏枯草(PRUNELLAE SPICA): 半枝莲(SCUTELLARIAE BARBATAE
HERBA): 牛樟芝(ANTRODIA CAMPHORATA):姜黄(CURCUMAE RHIZOMA): 牡丹皮(PAEONIAE SUFFRUTICOSAE
CORTEX): 败酱草(PATRINIAE HERBA): 小金英(IXERIS HERBA): 桑黄(PHELLINUS LINTEUS)=(0.5~5.0):
(0.5~5.0):(0.5~5.0): (0.5~5.0): (0.5~5.0): (0.5~5.0): (0.5~5.0):(0.5~5.0)。本发明的组合物,其剂量对于须治疗的哺乳类可为每天0.4克至120克。本领域技术人员需依个人的体重及病情而定,对于指定的个人可增减各剂量比例以实现最佳的效果。更具体而言,对于指定的组合物的剂量,范围可为每天0.4克至100克,较优地为每天10克至25克。较佳地,此等组合物至少每天使用3次,并且,大丸剂用药有效。应了解的是本文所述剂量适用于以干燥形式的草药(萃取物附着于研磨的植物或吸附剂上)。另外,本发明组合物的萃取物会增加活性物的浓度,因此要相应降低剂量。低至涵盖本发明组合物所述的10%的剂量。
本发明组合物较优地以口或肠内给药,然而,静脉内(I.V.)及/或肌肉内(I.M.)给药,亦可使用。本领域技术人员要了解如何制备I.V.及I.M.调配物,及如何可得到有效剂量。
用为本发明的起始物质的药草可得自商业基源,为单药草草药,可将其混合或萃取及浓缩,并置于组合物中以供人类使用。植物萃取物,一旦自植物材分离,可浓缩再制备为可供人类使用的形式(亦即丸剂、胶囊及锭剂)。
本发明所述医药产品通过以下步骤制得:(a)将粉碎的植物材料混合物与水接触形成水分散液;(b)加热水分散液至约100℃并维持于该温度约0.5小时至约3小时;(c)自水相分离不溶的植物材料;及(d)将含于水相中的溶质浓缩。通过冷冻干燥、喷雾干燥、蒸发或超滤而得到浓缩的溶质。
上述中草药及药草的特殊说明可于下面参考中找到:
(1)甘伟松:药用植物学,中国医药研究所,台北,中国台湾1973: 271,485,532,641,642,643。(2)林景彬:常用中药药理与应用(一),中国医药学院, 台中,中国台湾1985;201~207,236~246 (1985)。(3)邱年永与张光雄:原色台湾药用植物图鉴(1),南天书局,台北,中国台湾1995;186,188。(4)邱年永与张光雄:原色台湾药用植物图鉴(2),南天书局,台北,中国台湾1995;245。(5)邱年永与张光雄:原色台湾药用植物图鉴(4),南天书局,台北,中国台湾1995;81。(6)张宪昌:药草(一),渡假出版社有限公司,台北,中国台湾1997: 15。(7)张宪昌: 药草(二),渡假出版社有限公司,台北,中国台湾1997: 15。(8)李沐勋与李威着:常用中草药手册,中国医药研究所,台北,中国台湾2001:30, 44, 62, 85,
98,103,133,142。
要注意的是于传统中药实务中,草药用来治疗患者的症状,而非疾病或病原物本身,并且并非对特别疾病有特异性。草药依个别患者的症状而处方各异。草药的组合物亦随病例而异,甚至于治疗过程中,依各治疗结果,需对个别患者加以改变配方。因此很难从现有技艺中配方出一种适宜特殊疾病的特别的药草组合物。本发明的治疗癌症的药草组合物为得自个别药草、药草组合物及商业上可得的中草药。此等新颖的药草组合物及其萃取物于本文证明具有毒杀各种癌细胞的活性,并且可明显地增强其他抗癌药物效果。
本申请通过可利用的数据、完全描述物质的组合物、以及其制备方法、临床应用及将各种活性组成份特定化作为分析工具,通过以下实施例说明本发明的上述效果,但以下实施例并不用于限定本发明的保护范围。
附图说明
图1显示分别以0.5 mg / ml, 1 mg / ml, 5 mg / ml及10 mg / ml浓度的药草组合物萃取物处理后,黑色素瘤细胞A375的存活率。
图2显示分别以0.5 mg / ml, 1 mg / ml, 5 mg / ml及10 mg / ml浓度的药草组合物萃取物处理后,前列腺癌细胞PC3的存活率。
图3显示分别以0.5 mg / ml, 1 mg / ml, 5 mg / ml及10 mg / ml浓度的药草组合物萃取物处理后,大肠癌细胞HT29的存活率。
图4显示分别以0.5 mg / ml, 1 mg / ml, 5 mg / ml及10 mg / ml浓度的药草组合物萃取物处理后,肺癌细胞A549的存活率。
图5显示分别以0.22 mg / ml, 0.44 mg / ml, 0.87 mg/ ml, 1.75 mg / ml, 3.50
mg / ml及7.00 mg / ml浓度的药草组合物萃取物处理后,胃癌细胞AGS的存活率。
图6显示分别以0.24 mg / ml, 0.48 mg / ml, 0.95 mg / ml, 1.90 mg / ml,3.80
mg / ml 及7.60 mg / ml浓度的药草组合物萃取物处理后,乳癌细胞MCF-7的存活率。
图7显示分别以0.25 mg / ml, 0.51 mg / ml, 1.01 mg / ml, 2.02 mg / ml, 4.05
mg / ml及8.09 mg / ml浓度的药草组合物萃取物处理后,肝癌细胞Huh7的存活率。
图8显示以5 mg / ml的中草药组合物萃取物(有/无)用不同化疗药物处理后,多重抗药性大肠癌细胞HT29的存活率。
附图符号说明
细胞培养基:
C1-A:含有10 % FBS (fetal bovine serum胎牛血清), 100 units / ml
Penicillin(盘尼西林)及100 ug / ml Streptomycin(链霉素)的DMEM。
C1-B:含有10 % FBS (fetal bovine serum胎牛血清), 100 units / ml Penicillin及100 ug / ml Streptomycin的RPMI 1640。
C1-C: 含有10 % FBS (fetal bovine serum胎牛血清), 100 units / ml Penicillin及100 ug / ml Streptomycin的MEM。
C1-D:含有10 % FBS (fetal bovine serum胎牛血清), 100 units / ml Penicillin及100 ug / ml Streptomycin的F-12K。
阴性对照组:只有培养基而不加药。
阳性对照组:
Triton X100(聚乙二醇辛基苯基醚)(1 %),标示为C2。Triton X100是一种清洁剂,能杀死90 %以上的细胞。
S1:本药草组合物5 mg / ml
S2:5-FU(5-氟尿嘧啶)5 ug / ml
S3: Oxaliplatin(奥沙利铂) 10 ug / ml
S4: Irinotecan(伊立替康) 5 ug / ml
S5:(本药草组合物5 mg / ml与5-FU 5 ug / ml)
S6:(本药草组合物5 mg / ml与Oxaliplatin 10 ug / ml)
S7:(本药草组合物5 mg / ml与 Irinotecan 5 ug / ml)
S8:(本药草组合物5 mg / ml与5-FU 5 ug / ml与Oxaliplatin 10 ug / ml)
S9:(本药草组合物5 mg / ml与5-FU 5 ug / ml与Irinotecan 5 ug / ml)
S10:(5-FU 5 ug / ml与Oxaliplatin 10 ug / ml)
S11:(5-FU 5 ug / ml与Irinotecan 5 ug / ml)。
具体实施方式
为使本领域技术人员了解本发明的原理和效果,提出以下实施例,但仅作为例子,而不用于限定本发明的保护范围。
实施例1:药草组合物的制备-在制备本发明的药草组合物中,原材料为商业来源的以粉末形式存在的单药草型中草药。将各种单药草草药以相同重量混合制备各药草组合物。要强调的是,以个别单药草草药的来源植物制备的煎剂混合物或以各药草混合物中的单药草组成份与预先混合的来源植物制备的煎剂,也属于本发明的保护范围。
实施例2:单药草草药的制备-各单药草草药的植物来源公开于本申请背景技术中。应了解的是多于一个种或属的药用植物可用来制备相同的草药。例如夏枯草(PRUNELLAE SPICA)的制备来源可为滁州夏枯草(Prunella vulgaris)或日本夏枯草(Prunella vulgaris subsp.
asiatica)(亦称为Prunella vulgaris vra.
lilachina) 。半枝莲(SCUTELLARIAE BARBATAE
HERBA)的制备来源,可为半枝莲(Scutellaria barbata)、半枝莲(Scutellaria rivularis)或狗牙半枝莲(Scutellaria dependens)。牡丹皮(PAEONIAE SUFFRUTICOSAE
CORTEX)的制备来源可为牡丹(Paeonia suffruticosa)、矮牡丹(Paeonia suffruticosa var.
spontanea)、紫斑牡丹(Paeonia papaveracea)、川牡丹(Paeonia szechuanica)、黄牡丹(Paeonia lutea)、野牡丹(Paeonia delavayi)、保氏牡丹(Paeonia potanini)、茂纹牡丹(Paeonia thalictrumifolia)与云南牡丹(Paeonia yunnanonsis) 。草药从其各自的植物源制备方法如下:将部分或全草,以冷水洗,干燥及粉碎。植物材料再以沸水萃取,1份重量的植物材料对约5份至10份重量的水。所用水量至少盖住萃取容器中的植物材料。将样品煮沸0.5小时至1小时,但不超过3小时,以有效地萃取所需组成份。较短或较长的加热时间实质上不影响萃取,除了产量及费用外。通过过滤从植物材料中分离水溶液。通过冷冻干燥、喷雾干燥或加热,降低水溶液体积(真空或非真空环境下皆可)。较优的,可将浓缩物再喷雾干燥或冷冻干燥或吸收于相同植物材料、淀粉或其他吸收剂的粉末上。以此制备单药草草药。煎剂为植物材料的水溶液,通过将植物材料于水中煮沸约0.5小时至1小时而制得。煎剂于制备及冷却至适当温度后可直接服用或以适当灭菌保存供以后来服用。灭菌可通过微过滤或加热完成。
药草组合物的水萃取物对癌细胞的毒杀试验效果通过MTT assay鉴定。MTT assay:一种常被用于分析细胞存活率(percent of viable cells)及细胞毒性(cytotoxicity)的方法。MTT是一种 terazolium salt(四唑盐), 全名为3-[4,5-dimethylthiazol-2-yl]2,5-diphenyltetrazolium
bromide,为黄色染剂,可被活细胞吸收并被粒腺体中的琥珀酸四唑还原酶(succinate tetrazolium
reductase)还原 成蓝紫色的formazan结晶体,通过formazan的形成,即可计算细胞的存活率。细胞存活率(%) =(Nt / Nc) X 100。Nt是加药组的吸光值,而Nc是未加药组(阴性对照组)的吸光值。阴性对照组:只用细胞培养基,其细胞存活率定为100 %。以不同浓度的药物为横轴且以细胞存活率为纵轴而绘制柱状图。
细胞培养基:
C1-A:含有10 % FBS (fetal bovine serum胎牛血清), 100 units / ml
Penicillin及100 ug / ml Streptomycin的DMEM;
C1-B:含有10 % FBS (fetal bovine serum胎牛血清), 100 units / ml
Penicillin及100 ug / ml Streptomycin的RPMI 1640;
C1-C: 含有10 % FBS (fetal bovine serum胎牛血清), 100 units / ml
Penicillin及100 ug / ml Streptomycin的MEM;
C1-D:含有10 % FBS (fetal bovine serum胎牛血清), 100 units / ml
Penicillin及100 ug / ml Streptomycin的F-12K。
MTT assay-A:将癌细胞于培养基中培养2天后,于96孔盘培养皿中每孔殖入10,000个癌细胞,培养24小时后,分别以各种不同浓度的本药草混合物处理。处理之后, 在无血清的培养基内细胞与100 µl 的 MTT (0.5 mg/ml)共同培养4小时,之后将由存活细胞所产生的formazan结晶体以100 µl solubilisation
buffer(缓冲液) (10 % (v/v) Triton X-100 与 0.1N HCl in isopropanol)处理1小时而溶解。在波长570 nm下以Beckmann Coulter Elisa
plate reader (BioTek Power Wave XS) 测量溶液的光密度,样品经三次分析且以上述的公式计算细胞相对于阴性对照组(未加药组)的存活率(%)。只有培养基而不加药的培养皿当阴性对照组。Triton X100是一种清洁剂,能杀死90 %以上的细胞,Triton X100 (1 %)当阳性对照组,标示为C2。
MTT assay-B:MTT assay-B与MTT assay-A的原理是相同的且其方法是相似的。但是MTT assay-B以SpectraMax M5 Microplate
Reader (Molecular Devices)测量溶液的光密度。此外本检测方法无阳性对照组。阴性对照组:只用细胞培养基,其细胞存活率定为100 %,未标示于图中。
实施例3:药草组合物的水萃取物对人类恶性黑色素肿瘤细胞A375的毒杀试验。人类恶性黑色素肿瘤细胞A375的细胞培养基:C1-A。将人类恶性黑色素肿瘤细胞A375分别以0.5 mg / ml、1 mg / ml、5 mg / ml及10 mg / ml浓度的实施例1的组合物处理后,以MTT assay-A来测定细胞的存活率。阴性对照组:C1-A,阳性对照组:C2。实验结果如图1所示,其浓度相关性为,随着本发明组合物的浓度递增其毒杀癌细胞的活性亦递增。且依照实验结果计算其IC 50为3.20 mg / ml。
实施例4:药草组合物的水萃取物对人类前列腺癌细胞PC3的毒杀试验。人类前列腺癌细胞PC3的细胞培养基:C1-A。将人类前列腺癌细胞PC3分别以0.5 mg / ml、1 mg / ml、5 mg / ml及10 mg / ml浓度的实施例1的组合物处理后,以MTT assay-A来测定细胞存活率。阴性对照组:C1-A,阳性对照组:C2。实验结果如图2所示:本发明组合物的浓度为1 mg / ml ~10 mg / ml时,其浓度相关性为:随着本发明组合物的浓度递增其毒杀癌细胞的活性亦递增。且依照实验结果计算其IC 50为8.36 mg / ml。
实施例5:药草组合物的水萃取物对人类大肠癌细胞HT29的毒杀试验。人类大肠癌细胞HT29的细胞培养基:C1-B。将人类大肠癌细胞HT29分别以0.5 mg / ml、1 mg / ml、5 mg / ml及10 mg / ml浓度的实施例1的组合物处理后,以MTT assay-A来测定细胞存活率。阴性对照组:C1-B,阳性对照组:C2。实验结果如图3所示:本发明组合物的浓度为1 mg / ml ~10 mg / ml 时,其浓度相关性为:随着本发明组合物的浓度递增其毒杀癌细胞的活性亦递增。且依照实验结果计算其 IC 50为3.47 mg / ml。
实施例6:药草组合物的水萃取物对人类肺癌细胞A549的毒杀试验。
人类肺癌细胞A549的细胞培养基:C1-C。将人类肺癌细胞A549分别以0.5 mg / ml、1 mg / ml、5 mg / ml及10 mg / ml浓度的实施例1的组合物处理后,以MTT assay-A来测定细胞存活率。阴性对照组:C1-C,阳性对照组:C2。实验结果如图4所示其浓度相关性为:随着本发明组合物的浓度递增其毒杀癌细胞的活性亦递增。虽然在5 mg / ml浓度时肺癌细胞A549的存活率只剩34%,然而依照实验结果计算其IC 50为5.04 mg / ml。
实施例7:药草组合物的水萃取物对人类胃癌细胞AGS的毒杀试验。
人类胃癌细胞AGS的细胞培养基:C1-D。将人类胃癌细胞AGS分别以0.22 mg / ml、0.44 mg / ml、0.87 mg / ml、1.75 mg / ml、3.50 mg / ml及7.00 mg / ml浓度的实施例1的组合物处理后,以MTT assay-B来测定细胞的存活率。实验结果如图5所示,其浓度相关性为:随着本发明组合物的浓度递增其毒杀癌细胞的活性亦递增。且依照实验结果计算其 IC 50为3.19 mg / ml。
实施例8:药草组合物的水萃取物对人类乳癌细胞MCF-7的毒杀试验。
人类乳癌细胞MCF-7的细胞培养基: C1-C。将人类乳癌细胞MCF-7分别以0.24 mg / ml、0.48 mg / ml、0.95 mg / ml、1.90 mg / ml、3.80 mg / ml及7.60 mg / ml浓度的实施例1所述的组合物处理后,以MTT assay-B 来测定该细胞的存活率。实验结果如图6所示:本发明组合物的浓度为0.95 mg / ml ~7.60 mg/ml 时,其浓度相关性为:随着本发明组合物的浓度递增其毒杀癌细胞的活性亦递增。且依照实验结果计算其IC 50为4.38 mg / ml。
实施例9:药草组合物的水萃取物对人类肝癌细胞Huh7的毒杀试验。
人类肝癌细胞Huh7的细胞培养基:C1-A。将人类肝癌细胞Huh7分别以0.25 mg / ml、0.51 mg / ml、1.01 mg / ml、2.02 mg / ml、4.05 mg / ml及8.09 mg / ml浓度的实施例1的组合物处理后,以MTT assay-B来测定细胞存活率。实验结果如图7所示,其浓度相关性为:随着本发明组合物的浓度递增其毒杀癌细胞的活性亦递增。且依照实验结果计算其IC 50为1.50 mg / ml。
实施例10:药草组合物的水萃取物与其他化疗药物并用对大肠癌细胞HT29的毒杀试验。5-FU (5-fluorouracil)购自Celon Lab,Irinotecan 购自Dr. Reddy Lab ,Oxaliplatin 购自Panacea Biotech。人类大肠癌细胞HT29的细胞培养基: C1-B。将人类大肠癌细胞HT29分别以S1:本药草组合物5 mg / ml、S2:5-FU 5 ug / ml、S3:Oxaliplatin 10 ug / ml、S4: Irinotecan5 ug / ml、S5:(本药草组合物5 mg / ml与5-FU 5 ug / ml)、S6:(本药草组合物5 mg / ml与Oxaliplatin 10 ug / ml)、S7:(本药草组合物5 mg / ml与 Irinotecan 5 ug / ml)、S8:(本药草组合物5 mg / ml与5-FU 5 ug / ml与Oxaliplatin 10 ug / ml )、S9:(本药草组合物5 mg / ml与5-FU 5 ug / ml与Irinotecan 5 ug / ml)、S10:(5-FU 5 ug / ml与Oxaliplatin 10 ug / ml )及S11:(5-FU 5 ug / ml与Irinotecan 5 ug / ml)处理后,以MTT assay-A来测定细胞存活率。阴性对照组:C1-B,阳性对照组:C2。由图8所示,本发明药草组合物确实有很好的毒杀癌细胞活性,并能与其他抗癌药物产生明显的协同作用, 因此,本发明的组合物确实可当作癌症治疗的辅佐剂以增强其他抗癌药物的效果。
本文所述的治疗作用,可通过以上方法制得的草药实现,或以茶、煎药、饮料、糖果或其他糖膏剂、肠内液体营养产品如婴儿配方及成人营养产品、药用食物、营养补充品或营养品等形式服用。对于药品药物,可以为单位剂型如胶囊、小包或锭剂(有或无控制释放涂膜)服用。本发明仅针对于一部分癌症,即发现特定的药用植物或草药或它们的混合物具有令人惊讶的抗癌活性。更具体地,本发明的组合物是由具有人类消费的悠久历史的各种中国草药或药用植物衍生的。它们是GRAS(通常认为是安全的)。
工业上应用性:化疗的副作用很多,诸如疲劳、反胃、便秘与腹泻、口腔溃烂、头皮与脱发、皮肤痕痒剥落、神经和肌肉麻痹、听觉改变、血液出现问题以及生育能力下降。放疗的副作用也很多,诸如疲劳、血液改变、口腔疼痛、味觉改变、吞咽困难、恶心和呕吐、食欲不振、体重下降、声音转变、便秘或腹泻、毛发脱落、皮肤痕痒剥落以及生育能力下降。上述诸副作用经常困扰着癌症患者且使其痛苦不堪。此外因化疗或放疗而被杀灭的正常细胞会产生WNT16B,该WNT16B会促进存活的癌细胞成长、转移与抵抗治疗。 参见Yu Sun et., Treatment-induced damage
to the tumor microenvironment promotes prostate cancer therapy resistance
through WNT16B, Sep 2012; 18(9): 1359~1368。而本发明组合物的8个单方中草药中有5个(夏枯草、牛樟芝、姜黄、小金英及桑黄)被中国台湾卫生主管机关认定可做为食品原料,另外3个单方中草药(半枝莲、牡丹皮及败酱草)则为自古以来中医师常用的药,故其安全性不言可喻。
依照Eur. J. Cancer教导,Professor Jean-Francois MD说:“对于大肠癌细胞,手术前不论有无并用化疗的放射疗法,其后以5- Fu为辅助化疗并不能影响其无病细胞存活率或整体细胞存活率。我们的试验并不支持手术前不论有无并用化疗的放射疗法其后辅助化疗的现行做法。结合新辅助化疗的新的治疗策略是必需的。”参见Prof Jean-François Bosset
MD et al., Fluorouracil-based adjuvant chemotherapy after preoperative
chemoradiotherapy in rectal cancer: long-term results of the EORTC 2292
randomised study, The Lancet Oncology, Feb 2014; 15(2); 184~190。而本发明组合物可明显地增强其他抗癌药物效果,将能满足其需求成为新的化疗辅助剂。
依照世界卫生组织2014年世界癌症报告,癌症在未来20年将成为全球健康照护很大的负担,其发生率将增加57 %,且其死亡率将增加63 %。全球对抗癌症之战,不能只用治疗赢得胜利。迫切需要有效的预防方法来预防癌症危机。参见Stewart BW, Wild CP,
editors (2014). World Cancer Report 2014。而本发明组合物既安全又有明显的毒杀癌细胞作用,所以既可预防癌症又可治疗癌症,正能满足如此迫切的需求,所以本发明组合物的工业应用性很明显。
医药研究团体经常需要研究可以预防癌症或治疗癌症或辅助治疗癌症的方法及产品。本发明的萃取物产品及萃取物将很容易地为医药团体接受作为预防癌症或治疗癌症或癌症治疗辅助剂的另一选择。并且,本发明组合物显示出可明显地增强其他抗癌药物效果。此外越来越多的人喜欢使用更温和的中国传统医学治疗癌症。
虽然本发明的较佳具体实施例已经公开,但本领域技术人员在其中做的各种改变及修饰仍不会偏离本发明的保护范围。
Claims (5)
1.一种用于预防癌症、治疗癌症及增强其他抗癌药物效果的组合物,由夏枯草(PRUNELLAE
SPICA)、半枝莲(SCUTELLARIAE
BARBATAE HERBA)、牛樟芝(ANTRODIA
CAMPHORATA)、姜黄(CURCUMAE
RHIZOMA)、牡丹皮(PAEONIAE
SUFFRUTICOSAE CORTEX)、败酱草(PATRINIAE HERBA)、小金英(IXERIS HERBA)及桑黄(PHELLINUS LINTEUS)的萃取物所组成一种植物水溶性萃取物。
2.如权利要求1所述的组合物,其特征在于:各组分重量比大约为夏枯草(PRUNELLAE SPICA):半枝莲(SCUTELLARIAE BARBATAE HERBA):牛樟芝(ANTRODIA CAMPHORATA):姜黄(CURCUMAE RHIZOMA): 牡丹皮(PAEONIAE SUFFRUTICOSAE CORTEX):败酱草(PATRINIAE HERBA): 小金英(IXERIS HERBA): 桑黄(PHELLINUS LINTEUS)=(0.5~5.0):(0.5~5.0):(0.5~5.0):(0.5~5.0):(0.5~5.0):(0.5~5.0):(0.5~5.0):(0.5~5.0)。
3.如权利要求1所述的组合物,其特征在于:该萃取物的有效量为每天0.4~120克。
4.如权利要求1所述的组合物,其特征在于:该组合物经口或经鼻或经眼或经耳或经皮肤或经注射或经肛门或经生殖器而给药或通过吸入喷雾或植入贮存器的方式或以手术等侵入性方式施用于内脏组织的患部而给药或以针炙穴位经络给药。
5.如权利要求1所述的组合物,其特征在于:其进一步包括选自以下所组成的群的添加物:糖果、甜点、凝胶、营养补充品、口香糖、医药及婴儿营养品、饮料、优格、牛奶、再水合溶液及水溶液。
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