CN106999470A - 千忧解药物在用于制备治疗癌症的医药组合物中的用途 - Google Patents

千忧解药物在用于制备治疗癌症的医药组合物中的用途 Download PDF

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CN106999470A
CN106999470A CN201580057095.3A CN201580057095A CN106999470A CN 106999470 A CN106999470 A CN 106999470A CN 201580057095 A CN201580057095 A CN 201580057095A CN 106999470 A CN106999470 A CN 106999470A
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陈丘泓
庄秀美
魏宗德
萧乃文
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Launx Biomedical Co ltd
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Abstract

本发明提供千忧解(盐酸度洛西汀)在制备治疗癌症的医药组合物中的用途。

Description

千忧解药物在用于制备治疗癌症的医药组合物中的用途 技术领域
本发明为千忧解药物的新适应症的应用，尤其该千忧解药物具有抑制多种癌症的用途。
背景技术
癌症长期高居全球死因之首，且罹患癌症人数更是逐年攀升，因此治疗癌症俨然成为重要的课题。癌症的治疗可区分为手术治疗、放射线治疗、化学治疗及标靶治疗。
一般来说，癌症药物治疗无论是化学治疗或标靶治疗，目的多是让癌细胞无法复制、分裂，来阻断肿瘤的蔓延扩张。
在癌症药物的设计上也是均朝向高专一性的药物分子开发或标靶型的抗体设计，根据统计，平均每一万个新药约只有五个能够进入第一期临床试验。
此外，除了药物本身是否能大量制造的难题外，还需克服药物安全性、病人筛选、试用剂量等问题，即便药物已经通过FDA的核准并上市，还是常遇到患者对于治疗药物的反应不佳。进一步，许多癌细胞相继产生抗药性，使药物的使用成效大幅降低，最终导致癌症治疗失败。由此可见药物开发具有一定的困难程度。
千忧解(Duloxetine HCl、盐酸度洛西汀)是一种为口服选择性血清素与正肾上腺素再吸收抑制剂(SSNRI)Duloxetine且于人体的抗忧郁作用机转未知，推测与其作用于中枢神经之血清素活性(serotonergic)与正肾上腺素活性 (noradrenergic)有关。臨床前试验显示duloxetine(度洛西汀)可有效阻断血清素与正肾上腺素的再吸收，而仅些微抑制多巴胺(dopamine)的再吸收。体外试验显示，Duloxetine对多巴胺性(dopaminergic)、肾上腺素性(adrenergic)、胆碱性(cholinergic)或组织胺性(histaminergic)接受器无显著亲合力。Duloxetine不抑制单胺氧化酶。Duloxetine经广泛代谢，但尚未发现其主要代谢物显著具有duloxetine的药理活性。千忧解是早已被FDA所认可的用药，具有大量人体研究成果资料。
由于临床应用上的差异，因此从来没有人认为千忧解具有抑制癌症细胞的可能性。
发明内容
为解决上述的问题，本发明针对千忧解药物进行新适应症的研发，而达到老药新用的目标。
经过实验设计结果显示千忧解药物对正常细胞没有或仅有微小的毒性，但至于千忧解在正常细胞与肿瘤细胞之间是否具有选择性的影响。
本发明提供一种千忧解(Duloxetine HCl)药物在用于制备治疗癌症的医药组合物中用途，其中所述医药组合物是选自由有效剂量的千忧解及药学上可接受的盐类所组成。
本发明一实施例中，其中所述癌症是选自由胸腔相关癌症、腹腔相关癌症、内分泌相关癌症、消化道相关癌症中的一种或多种。
本发明一实施例中，其中所述癌症是选自由骨肉瘤相关癌症、皮肤癌相关癌症、血癌相关癌症中的一种或多种。
本发明一实施例中，其中所述胸腔相关癌症是指肺癌。
本发明一实施例中，其中所述腹腔相关癌症是选自由膀胱癌或/和子宫颈癌。
本发明一实施例中，其中所述内分泌相关癌症是选自由前列腺癌、乳癌、卵巢癌中的一种或多种。
本发明一实施例中，其中所述内分泌相关癌症是选自由胃癌、肝癌、大肠癌、胰脏癌、舌癌中的一种或多种。
本发明一实施例中，其中所述千忧解药物的有效剂量浓度为20～500mg/kg/天。
附图说明
图1显示本发明千忧解用药应用于抑制癌细胞分析结果。
图2显示千忧解用药可对肿瘤体积的效果
图3显示高剂量及低剂量千忧解用药对肿瘤生长的抑制效果
具体实施方式
各种癌症细胞株建立
将不同癌症类型的细胞株进行继代培养，癌症细胞种类包含肺癌、胃癌、肝癌、直肠癌、皮肤癌、子宫颈癌、前列腺癌、膀胱癌、乳癌、血癌、胰腺癌、卵巢癌、舌癌、骨肉瘤、肾脏癌，并且对照组亦使用肾脏(HEK293(Kidney))、肺部上皮细胞BEAS-2B(Lung Epithelial)的正常细胞做测试。(表一)
先将各细胞于培养液培养后，由于各细胞的属性不同，因此针对不同种的细胞也要用相对应的培养液(表一)，计算细胞数目，回种2×106细胞数，然后加入培养该细胞株的培养液补至体积为10ml，继续培养2-3天。之后将细胞计数，并分装至96孔盘，其中每孔的细胞数目固定为3000颗，且体积为100ul。
表一、癌症测试细胞株及其培养所用的培养液
细胞存活分析方法
将96孔盘中原有的培养液吸掉，每孔加入浓度10um、体积100ul的市售药物，放置72小时后，每孔再加入100ul已稀释的WST-1试剂，该稀释比例为培养液与WST-1原液的体积比为9：1，最后每个孔盘的总体积为200ul，然后将 96孔盘放置于37℃，30～90分钟，利用elisa reader(酶联免疫检测仪)于OD450侦测吸光值，并计算各癌症细胞株的存活率。
千忧解药对于各种癌细胞分析结果
千忧解对胸腔相关癌症细胞抑制效果的测试
本分析千忧解对胸腔相关癌症细胞抑制效果的测试，主要针对两种肺癌细胞进行测试，癌细胞株分别为A549和H1650两个细胞株，各做4次的癌细胞抑制实验结果，并且计算其平均值，结果表列如后。(表二)
表二、千忧解对肺癌相关癌症细胞抑制测试
  0524-10min 0526-10min 0529-10min 0531-10min 平均
A549 32.8 53.7 33.2 30.1 37.5
  1-10min 2-20min 3-20min 4-20min 平均
H1650 35.2 36.9 33.6 39.4 36.3
千忧解对腹腔相关癌症细胞抑制效果的测试
本分析千忧解对腹腔相关癌症细胞抑制效果的测试，主要针对两种腹腔相关癌症种类细胞进行测试，膀胱癌细胞株分别为TSGH和T24两个细胞株(表三)，子宫颈癌细胞株分别为HeLa细胞株(表四)，肾脏癌为786-O细胞株(表五)，各做4次的癌细胞抑制实验结果，并且计算其平均值，结果表列如后。
表三、千忧解对膀胱癌相关癌症细胞抑制测试
  0510-10min 0512-10min 0515-10min 0517-10min 平均
TSGH 36.1 59.0 61.2 40.5 49.2
  T24-1-30min T24-2-20min T24-3-20min T24-4-20min 平均
T24 81.7 90.6 71.4 97.2 85.2
表四、千忧解对子宫颈癌相关癌症细胞抑制测试
  0524-10min 0526-10min 0529-10min 0531-10min 平均
HeLa 27.7 29.9 25.4 52.1 33.8
C-334 34.3 52.4 33.7 42.4 40.7
表五、千忧解对肾脏癌相关癌症细胞抑制测试
  0524-10min 0526-10min 0529-10min 0531-10min 平均
786-O 60.4 89.7 49.0 70.1 67.3
千忧解对内分泌相关癌症细胞抑制效果的测试
本分析千忧解对腹腔相关癌症细胞抑制效果的测试，主要针对三种内分泌相关癌症种类细胞进行测试，前列腺癌细胞株分别为PC-3和LNCap两个细胞株(表六)，乳癌细胞株分别为MCF7和MDA-MB-231两个细胞株(表七)，卵巢癌细胞株分别为NIH-OVCAR-3细胞株和TOV-21G(表八)，各做4次的癌细胞抑制实验结果，并且计算其平均值，结果表列如后。
表六、千忧解对前列腺癌相关癌症细胞抑制测试
表七、千忧解对乳癌相关癌症细胞抑制测试
表八、千忧解对卵巢癌相关癌症细胞抑制测试
  7-3-30min 7-4-30min 7-7-30min -4-30min 平均
NIH-OVCAR-3 64.8 74.6 78.5 55.0 68.2
  7-3-30min 7-4-30min 7-7-30min -4-30min 平均
TOV-21G 30.9 41.9 34.7 31.9 34.9
千忧解对消化道相关癌症细胞抑制效果的测试
分析千忧解对消化道相关癌症细胞抑制效果的测试，主要针对五种消化道相关癌症种类细胞进行测试，胃癌细胞株分别为AGS和MKN-45两个细胞株(表九)，肝癌细胞株分别为HepG2和Hep3B两个细胞株(表十)，大肠癌细胞株分别为HCT116-wt和LoVo细胞株(表十一)，胰脏癌细胞株分别为AsPC和BxPC细胞株(表十二)，舌癌细胞株为SAS细胞株(表十三)，各做4次的癌细胞抑制实验结果，并且计算其平均值，结果表列如后。
表九、千忧解对胃癌相关癌症细胞抑制测试
  0510-10min 0512-10min 0515-10min 0517-10min 平均
AGS 25.0 15.9 37.3 15.8 23.5
  0510-10min 0512-10min 0515-10min 0517-10min 平均
MKN-45 59.5 75.3 46.0 57.9 59.7
表十、千忧解对肝癌相关癌症细胞抑制测试
  0524-20min 0526-20min 0529-20min 0531-20min 平均
HepG2 14.8 44.7 37.7 38.9 34.0
  0612-20min 0614-20min 0616-20min 0619-20min 平均
Hep3B 41.4 53.3 36.2 60.6 47.9
表十一、千忧解对大肠癌相关癌症细胞抑制测试
  0602-30min 0605-10min 0607-10min 0609-10min 平均
HCT116-wt 52.1 108.8 127.4 20.3 77.1
  0616-10min 0619-10min 0621-10min 0623-10min 平均
LoVo 44.8 63.4 49.9 31.0 47.3
表十二、千忧解对胰脏癌相关癌症细胞抑制测试
  1-7-3-30min 1-7-4-30min 1-7-7-30min 1-4-30min 平均
AsPC 75.7 78.0 26.0 61.7 60.3
  3-7-3-30min 3-7-4-30min 3-7-7-30min 3-4-30min 平均
BxPC 62.1 58.8 22.7 72.4 54.0
表十三、千忧解对舌癌相关癌症细胞抑制测试
  6-26-10min 6-28-10min 6-30-10min 7-3-10min 平均
SAS 45.7 88.9 96.2 98.9 82.4
千忧解对其他癌症细胞抑制效果的测试
分析千忧解对其他类型癌正进行测试，骨肉癌细胞株为U2OS细胞株(表十四)，皮肤癌细胞株分别为A375和BCC两个细胞株(表十五)，各做3～4次的癌细胞抑制实验结果，并且计算其平均值，结果表列如后。
表十四、千忧解对骨肉瘤相关癌症细胞抑制测试
  6-26-10min 6-28-10min 6-30-10min 7-3-10min 平均
U2OS 26.6 21.5 56.6 35.6 35.1
表十五、千忧解对皮肤癌相关癌症细胞抑制测试
对照组实验设计
千忧解对正常细胞抑制效果的测试
分析千忧解对多种正常细胞进行测试，正常肾脏细胞株为HEK293细胞株(表十六)，正常肺部上皮细胞细胞株为BEAS-2B细胞株(表十七)，各做4次的癌细胞抑制实验结果，并且计算其平均值，结果表列如后。
表十六、千忧解对正常肾脏细胞抑制测试
  平均
HEK293 62.4
表十七、千忧解对正常肺部上皮细胞抑制测试
  0510-10min 0512-10min 0515-10min 0517-10min 平均
BEAS-2B 59.5 76.3 74.7 79.6 72.5
针对千忧解对各种癌症细胞抑制效果汇整于表十八中，清楚看出千忧解对于多项癌症有明显的抑制效果。经过发明人实验结果，千忧解药物对于不同的癌症细胞有明显的抑制效果。(图一)
表十八、千忧解对各种癌症细胞抑制效果的汇整
癌症细胞 抑制效果
肺癌 36.8743253
膀胱癌 67.2
子宫颈癌 36.76
前列腺癌 59.23
乳癌 48.57
卵巢癌 51.5
胃癌 41.6
肝癌 40.95
大肠癌 62.21
胰脏癌 57.2
舌癌 82.44
骨肉癌 35.09
皮肤癌 39.79
肾脏癌 67.3
肾脏 62.44
肺部上皮细胞 72.50
动物实验分析
本实验使用雌性BALB/cAnN.Cg-Foxn1nu/CrlNarl小鼠为样本(购自国家实验动物中心)，体重为21±1g，皮下注射肝癌细胞(HepG2)后随机分笼，测试药物分成三组:正常对照组、低剂量(100mg/kg/day)、高剂量(200mg/kg/day)。肿瘤形 成超过100mm3后，每天采取腹腔注射方式给予药物；每周测量肿瘤大小两次，肿瘤体积测量公式如下：(L×W2)；L代表肿瘤最长直径；W代表肿瘤最短直径。
依据第2图的结果，低剂量与高剂量的千忧解均对肿瘤具有良好的抑制效果，且于实验过程中各组小鼠的重量均未出现明显降低的现象，因此表示千忧解不论高低剂量在治疗过程均能使受测小鼠具有良好的健康状态而不死亡。
依据第3图的结果，高低剂量的千忧解平可有效减缓肿瘤体积增长，并可同时减少肿瘤体积，其中以高剂量的千忧解具有较佳的效果。
上列详细说明是针对本发明之一可行实施例的具体说明，惟该实施例并非用以限制本发明的专利范围，凡未脱离本发明技艺精神所为的等效实施或变更，均应包含于本发明的专利范围中。

Claims (8)

  1. 一种千忧解药物在用于制备治疗癌症的医药组合物中的用途，其特征是，所述医药组合物是选自由有效剂量的千忧解及药学上可接受的盐类所组成。
  2. 如权利要求1所述的用途，其特征是，所述癌症为胸腔相关癌症、腹腔相关癌症、内分泌相关癌症、消化道相关癌症中的一种或多种。
  3. 如权利要求1所述的用途，其特征是，所述癌症是选自骨肉瘤相关癌症、皮肤癌相关癌症、血癌相关癌症中的一种或多种。
  4. 如权利要求2所述的用途，其特征是，所述胸腔相关癌症是指肺癌。
  5. 如权利要求2所述的用途，其特征是，所述腹腔相关癌症是选自膀胱癌或/和子宫颈癌。
  6. 如权利要求2所述的用途，其特征是，所述内分泌相关癌症系选自由前列腺癌、乳癌、卵巢癌中的一种或多种。
  7. 如权利要求2所述的用途，其特征是，所述消化道相关癌症系选自由胃癌、肝癌、大肠癌、胰脏癌、舌癌中的一种或多种。
  8. 如权利要求1所述的用途，其特征是，所述有效剂量浓度为20～500mg/kg/天。
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