CN102112141B - 用于预防前列腺癌或减缓前列腺癌发展的组合物 - Google Patents
用于预防前列腺癌或减缓前列腺癌发展的组合物 Download PDFInfo
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Abstract
描述的是一种组合物，其包括作为有效成分的绿茶提取物和石榴提取物，用于预防前列腺癌或者减缓前列腺癌发展。
Description
发明领域
本发明涉及用于预防前列腺癌或减缓前列腺癌发展的组合物。
具体地，本发明涉及组合物，其包括绿茶和石榴提取物作为有效成分，用于预防前列腺癌或者减缓前列腺癌发展。
发明背景
在哺乳动物中前列腺癌是最普遍的癌症，尤其在西方国家中的男性中(Cancer Statistics 1997，CA Cancer J.Clin.1997；47；5-27)。
各种因素诸如未知的病原学、易变的病理学、与内分泌因素的复杂关系、和退行性进程(anaplastic progression)导致该疾病的复杂性。
威斯康辛州麦迪逊市的威斯康辛大学和俄亥俄州克利夫兰市的凯斯西保留地大学的一组研究人员在人前列腺癌的小鼠模型中证明在调节前列腺肿瘤细胞中胰岛素样生长因子-1(IGF-1)驱动的分子途径中绿茶多酚(GTP)的作用。基于研究的这些观察结果具有重要性，表明增加的IGF-1水平如何与若干癌症诸如前列腺癌、乳腺癌、肺癌和结肠癌的增加风险相关。通过控制前列腺癌小鼠模型的血清中血管内皮生长因子(VEGF)的量，绿茶多酚有利于最小化肿瘤发展。VEGF的减少可以由GTP-引发的IGF-I水平的抑制引起。VEGF的作用是补充和发展向生长的肿瘤运送营养成分的新血管。通过减少VEGF的量，GTP起着将流向和支持肿瘤生长的营养成分最小化的作用。
在Clinical Cancer Research(临床癌症研究)，July 1，2006；pp 4018-4026和Clinical Cancer Research(临床癌症研究)Vol.12，4018-4026，July 1，2006中，据报道石榴提取物可以用于预防前列腺癌或者减缓前列腺癌发展。
在Prostate Cancer and Prostatic Diseases(前列腺癌和前列腺疾病)(2002)5，6-12中，据报道番茄红素可以用于预防前列腺癌或者减缓前列腺癌发展。
在WO0057892中，据报道蓝棕属可用于前列腺癌的治疗。
在WO03035635中，据报道异黄酮衍生物可用于前列腺癌的治疗；
在WO04091602中，据报道L-肉碱可用于心血管病的治疗。
人体中的硒是一种微量元素营养成分，其作用是作为减少抗氧化酶诸如谷胱甘肽过氧化酶的辅因子。膳食的硒来自坚果、谷类、肉、鱼和蛋。“营养预防癌症项目”(NPC)是一个控制的、随机化的癌症预防试验，其中1,312名患者接受每日200μg剂量的硒或者安慰剂长达10年。在该研究中，获得了前列腺癌发生率在统计学上显著的减少和前列腺癌发展在统计学上显著的减缓。
尽管绿茶和石榴提取物的抗癌作用已是已知的，但没有一个上述引用的现有技术文件提及或者建议这两种有效成分组合用于预防前列腺癌或者减缓前列腺癌发展。
此外，虽然有其他可利用的药剂用于肿瘤的化疗，和对于前列腺癌存在其他侵入性治疗选择，诸如切除癌变的前列腺或者放置设计用于收缩肿瘤的放射性粒源，但更期望提供一种对患者低毒性的可用作传统治疗方法的辅助或补充的组合物，其作为前列腺癌的抗癌药剂。
发明内容
因此，本发明的一个目标是一种增效组合物，其包括作为有效成分的绿茶提取物和石榴提取物。
上述组合物还可以包括番茄红素、硒、锌、蓝棕属、异黄酮衍生物和L-肉碱。
本发明的进一步目标是一种组合物，其包括：
(a)绿茶提取物，作为有效成分，以从25至800mg的剂量，优选的剂量是125和250mg；和
(b)石榴提取物，作为有效成分，以从25至800mg的剂量，优选的剂量是40、125和250mg；和
(c)番茄红素，以从0.03至30.0mg的剂量，优选的剂量是1.25和5mg；
(d)硒，以从8.2至500μg的剂量，优选的剂量是55和82.5g；
(d)锌，以从1至200mg的剂量，优选的剂量是20mg；和
(e)蓝棕属，以从10至400mg的剂量，优选的剂量是160和320mg；
(f)异黄酮衍生物(大豆异黄酮)，以从10至500mg的剂量，优选的剂量是100mg；和
(g)L-肉碱，以从50至500mg的剂量，优选的剂量是200mg。
本发明的进一步目标是上述组合物在预防前列腺癌或者减缓前列腺癌发展中的用途。
本发明的进一步目标是上述组合物用于制备预防前列腺癌或者减缓前列腺癌发展的药物的用途。
本发明的进一步目标是上述组合物用于制备预防前列腺癌或者减缓前列腺癌发展的膳食补充剂的用途。
本发明的组合物还可以包括辅酶、矿物质、抗氧化剂、维生素和可用于治疗前列腺癌的药剂。
下列非限制性的实施例进一步阐明本发明。
实施例1
对于在下列报告的实验，使用已确定的人类前列腺癌细胞系LNCaP和PC3(从American Type Culture Collection(美国典型培养物保藏中心)Rockville，MD获得)。
在RPMI 1640培养基(Life Technologies，Rockville，MD)中培养LNCaP细胞，以10％胎牛血清(FBS)和1％青霉素和链霉素补充。5％CO2在空气中的潮湿环境中在37℃培养细胞。两组细胞培养物在10cm组织培养瓶中生长至80％融合并且分裂1∶8。细胞被铺在96孔板中并且使其附着并达到60-70％融合，之后用于实验。
对在体外细胞生长的影响
以绿茶提取物(PBS中40μg/ml)、石榴提取物(PBS中40μg/ml)或者组合(绿茶提取物和石榴提取物)，在每一个包含完全细胞培养基的10个孔中处理培养的LNCaP和PC3细胞组。以相同量的PBS在完全细胞培养基中温育用作对照的细胞。通过将它们单独地或者与绿茶提取物和石榴提取物组合地加入细胞培养物，也测试番茄红素(5mmol/L)、硒(30ng/ml)、锌(10μg/ml)、蓝棕属(10μg/ml)和大豆异黄酮(10μg/ml)的影响。在37℃下24小时的温育之后，，用或者不用测试试剂，采用3-(4，5-二甲基噻唑-2-基)-2，5-二苯基四氮唑
(MTT)比色测定，确定细胞的细胞增殖。向所有的孔加入MTT(5ng/ml)并且再温育2小时。然后在4℃下以500xg将板离心5分钟。通过小心的吸出从孔移除MTT溶液，并向每一个孔加入缓冲的DMSO(0.1ml)并且晃动板15分钟。在全自动定量酶标仪上测量540nm波长处的吸光度。以细胞存活能力的百分比评价每种化合物单独地和组合地对细胞生长的影响，其中未经处理的对照细胞被认为是全部存活的并且被允许自由生长。
在下列表1-4中报告所得结果。
表1
测试化合物对LNCaP细胞增殖的影响
A＝绿茶提取物；B＝石榴提取物；C＝番茄红素；D＝硒；E＝锌。
表2
测试化合物对PC3细胞增殖的影响
表3
测试化合物对LNCaP细胞增殖的影响
A＝绿茶提取物；B＝石榴提取物；F＝蓝棕属；G＝大豆异黄酮；H＝L-肉碱
表4
测试化合物对PC3细胞增殖的影响
A＝绿茶提取物；B＝石榴提取物；F＝蓝棕属；G＝大豆异黄酮；H＝L-肉碱
表1、2、3和4报告的结果表明本发明的组合物相对于单一成分在统计上更有活性。
而且，番茄红素、硒和锌、或者蓝棕属、大豆异黄酮和L-肉碱的存在没有增加本发明组合物的抑制活性。
实施例2
实验2对体外细胞分裂的影响
用以下任一个处理25ml培养瓶中的LNCaP和PC3细胞：绿茶提取物(PBS中40μg/ml)、石榴提取物(PBS中40μg/ml)、蓝棕属(10μg/ml)、大豆异黄酮(10μg/ml)或者绿茶提取物加石榴提取物的组合。
对照只接受PBS。附着之后48小时开始处理。在使用流式细胞术分析之前，允许细胞在培养基中生长多达72小时。用10mM溴代脱氧尿苷(BrdU)脉冲标记细胞2小时，之前有或者没有用绿茶提取物或者石榴提取物或者二者处理非同步生长的细胞。然后收获细胞，使用70％乙醇固定，以0.1％HCl处理并且在90℃加热10分钟以暴露该标记的DNA。用抗-BrdU结合的FITC(Becton Dickinson)将细胞染色并且用碘化丙锭复染色。在Becton-Dickinson FACScan上使用Lysis II软件进行细胞周期分析。
下列表中报告，使用本发明组合物相对于单一成分获得的流式细胞术分析的结果。
表5
绿茶提取物，LNCaP细胞分裂BrdU
表6
石榴提取物，LNCaP细胞分裂BrdU
表7
蓝棕属提取物，LNCaP细胞分裂BrdU
表8
大豆异黄酮提取物，LNCaP细胞分裂BrdU
表9
绿茶加石榴提取物，LNCaP细胞分裂BrdU
表10
绿茶提取物PC3细胞分裂BrdU
表11
石榴提取物PC3细胞分裂BrdU
表12
蓝棕属提取物PC3细胞分裂BrdU
表13
大豆异黄酮提取物PC3细胞分裂BrdU
表14
绿茶加石榴提取物PC3细胞分裂BrdU
使用单一化合物表明对LNCaP和PC3两种细胞增殖的抑制作用(见表5-8和10-13)。表9和14中报告的结果表明本发明组合物被赋予抑制LNCaP和PC3两种细胞的增殖的意想不到的增效作用。细胞系停滞在S阶段中持续72个小时，在LNCaP和PC3系中分别为60.9％和54.8％的最大值。
在仅用载体处理的对照中没有观察到对细胞分裂或者生长停滞的影响。
实施例3
体内抗肿瘤活性
为评价本发明组合物的抗肿瘤活性，使用实体瘤菌株LNCaP(ATCC)。
在裸小鼠中培养细胞五至六次。这种菌株的一个实体癌片段(3mm2)被移植至雄性8周龄裸小鼠(NxGen Biosciences，San Diego，CA)的腋区皮肤下。(大约20天之后)可靠地拿掉肿瘤的小鼠被随机地分为4组，每组10只动物。以载体、绿茶提取物、石榴或者绿茶和石榴的组合，治疗动物组。对照组中的动物只接受饮用水，而绿茶和石榴提取物或者其组合以饮用水中1％(w/v，随意)每天一次在早上给药，连续28天。每周两次测量肿瘤直径，并且使用式0.5238L1L2H计算肿瘤体积，其中L1是长的直径，L2是短的直径和H是肿瘤的高度。在对照动物中一旦肿瘤达到大约1,300mm3，所有的动物被施以无痛致死术，这是在大约28天之后。
结果(平均和SE)表明对照组中在28天时肿瘤体积是1305±87，而在绿茶和石榴提取物治疗过的组中体积值分别是1026±98(p＜0.05)和1011±91(p＜0.05)。而在组合治疗中(绿茶加石榴提取物)，对肿瘤减小有确定无疑的增效作用，623±112(p＜0.001)。因此与仅绿茶(21.4％)或者石榴(22.5％)相比，组合物的抑制率是52.3％。
表15中报告的所得结果表明本发明的组合物相对于单一成分在统计学上更有活性。
表15
根据本发明的膳食补充剂或者药剂由医学领域的工作者熟悉并且已经用于临床实践的有效成分组成，并且其药物毒物学概况是已知的。
因此，它们的获得是很容易的，因为这些是目前已在市场上存在很长时间的产品并且具有适合人类或者动物给药的等级。
在下面，报告的是根据本发明的组合物的非限制性例子。
组合物1
绿茶提取物250mg；
石榴提取物250mg。
组合物2
绿茶提取物250mg；
石榴提取物250mg；
番茄红素1.25mg；
硒82.5μg；
锌20mg。
组合物3
绿茶提取物125mg；
石榴提取物125mg；
番茄红素5mg；
硒82.5μg；
锌20mg；
蓝棕属160mg；
大豆异黄酮100mg。
组合物4
绿茶提取物125mg；
石榴提取物40mg；
番茄红素5mg；
硒55μg；
锌20mg；
蓝棕属320mg；
大豆异黄酮100mg。
组合物5
绿茶提取物125mg；
石榴提取物125mg；
番茄红素5mg；
硒82.5μg；
锌20mg；
蓝棕属160mg；
大豆异黄酮100mg；
L-肉碱200mg。
Claims (5)
1.用于预防前列腺癌或者减缓前列腺癌发展的组合物，其有效成分为：25至800mg剂量的绿茶提取物、25至800mg剂量的石榴提取物、0.03至30.0mg剂量的番茄红素、8.2至500μg剂量的硒、1至200mg剂量的锌、10至400mg剂量的蓝棕属、10至500mg剂量的大豆异黄酮和50至500mg剂量的L-肉碱。
2.根据权利要求1所述的组合物，作为膳食补充剂或药物。
3.根据权利要求1-2任一项所述的组合物用于制备预防前列腺癌或者减缓前列腺癌发展的药物的用途。
4.25至800mg剂量的绿茶提取物和25至800mg剂量的石榴提取物联合用于制备预防前列腺癌或者减缓前列腺癌发展的膳食补充剂或药物的用途。
5.根据权利要求4的用途，在绿茶提取物和石榴提取物的基础上，进一步配伍0.03至30.0mg剂量的番茄红素、8.2至500μg剂量的硒、1至200mg剂量的锌、10至400mg剂量的蓝棕属、10至500mg剂量的大豆异黄酮和/或50至500mg剂量的L-肉碱。
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Also Published As
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|---|---|
| WO2010012651A3 (en) | 2010-04-01 |
| WO2010012651A2 (en) | 2010-02-04 |
| SG192527A1 (en) | 2013-08-30 |
| HRP20161447T1 (hr) | 2016-12-16 |
| PT2310023T (pt) | 2016-11-22 |
| KR101621910B1 (ko) | 2016-05-17 |
| CN102112141A (zh) | 2011-06-29 |
| KR20110043615A (ko) | 2011-04-27 |
| DK2310023T3 (en) | 2016-12-12 |
| EA201170269A1 (ru) | 2011-06-30 |
| HK1158076A1 (en) | 2012-07-13 |
| BRPI0916435A8 (pt) | 2017-12-26 |
| AU2009276035B2 (en) | 2015-10-01 |
| MY160546A (en) | 2017-03-15 |
| CA2732205A1 (en) | 2010-02-04 |
| JP5564499B2 (ja) | 2014-07-30 |
| JP2011529465A (ja) | 2011-12-08 |
| EP2310023A2 (en) | 2011-04-20 |
| BRPI0916435A2 (pt) | 2016-02-16 |
| US20110262417A1 (en) | 2011-10-27 |
| AU2009276035A1 (en) | 2010-02-04 |
| HUE031821T2 (en) | 2017-08-28 |
| US9084812B2 (en) | 2015-07-21 |
| EP2310023B1 (en) | 2016-10-12 |
| CA2732205C (en) | 2018-01-02 |
| EA019814B1 (ru) | 2014-06-30 |
| SG10201700334XA (en) | 2017-03-30 |
| PL2310023T3 (pl) | 2017-02-28 |
| MX2011000941A (es) | 2011-03-29 |
| ES2602834T3 (es) | 2017-02-22 |
| US20140255512A1 (en) | 2014-09-11 |
| US8758835B2 (en) | 2014-06-24 |
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