WO2016062287A1 - 山喜多药物在用于制备抑制癌症的医药组合物中的用途 - Google Patents

山喜多药物在用于制备抑制癌症的医药组合物中的用途 Download PDF

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WO2016062287A1
WO2016062287A1 PCT/CN2015/092777 CN2015092777W WO2016062287A1 WO 2016062287 A1 WO2016062287 A1 WO 2016062287A1 CN 2015092777 W CN2015092777 W CN 2015092777W WO 2016062287 A1 WO2016062287 A1 WO 2016062287A1
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cancer
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shan
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陈丘泓
庄秀美
魏宗德
萧乃文
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朗齐生物医学股份有限公司
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Definitions

  • the invention is an application of a new indication of a mountain medicinal drug, in particular, the mountain sedative drug has the use of inhibiting various cancers.
  • CellCept Mycophenolate mofetil, mycophenolate mofetil, mycophenolate mofetil is an immunosuppressive agent.
  • the main component is 2-morpholinoethyl ester (mycophenolate) of mycophenolic acid (MPA, mycophenolic acid).
  • MPA is a selective, non-competitive and reversible potent fiber nucleoside monophosphate dehydrogenase (IMPDH) inhibitor, thus inhibiting the denovo pathway of guanine nucleotide synthesis without binding to DNA, MPA pair Cytostatic effects caused by lymphocytes are stronger than those of other cells.
  • IMPDH potent fiber nucleoside monophosphate dehydrogenase
  • CellCept is effective in preventing organ rejection in patients undergoing allogeneic kidney transplantation, and in treating uncontrolled renal rejection and in preventing organ rejection in patients receiving allogeneic heart or liver transplantation.
  • Shan Xi Duo is a drug that has long been approved by the FDA, with a large number of drug machine transfers and human research results.
  • cancer has long been the leading cause of death worldwide, and the number of cancer patients has increased year by year. Therefore, treating cancer has become an important issue.
  • cancer drug treatment whether it is chemotherapy or target treatment, is intended to prevent cancer cells from replicating and dividing to block the spread and spread of tumors. According to statistics, only about five out of every 10,000 new drugs can enter the first phase of clinical trials. Further, many cancer cells are successive Drug resistance is produced, and the use of the drug is greatly reduced, eventually leading to failure of cancer treatment. This shows that drug development has a certain degree of difficulty.
  • the present invention aims at the development of new indications for the application of the drug, and achieves the goal of new use of the old drug.
  • the present invention provides a use of a Santoprene drug for the preparation of a pharmaceutical composition for inhibiting cancer, wherein the pharmaceutical composition comprises an effective amount of a sedative drug and a pharmaceutically acceptable salt.
  • the cancer is one or more selected from the group consisting of a chest-related cancer, an abdominal cavity-related cancer, an endocrine-related cancer, and a digestive tract-related cancer.
  • the cancer is one or more selected from the group consisting of an osteosarcoma-related cancer, a skin cancer-related cancer, and a blood cancer-related cancer.
  • chest cavity related cancer refers to lung cancer
  • the abdominal cavity-related cancer is selected from the group consisting of bladder cancer or/and cervical cancer.
  • the endocrine-related cancer is one or more selected from the group consisting of prostate cancer, breast cancer, and ovarian cancer.
  • the endocrine-related cancer is one or more selected from the group consisting of gastric cancer, liver cancer, colon cancer, pancreatic cancer, and tongue cancer.
  • the effective dose concentration of the Shanxi multi-drug is 20-500 mg/kg/ day.
  • Fig. 1 shows the results of the application of the present invention to inhibit cancer cell analysis.
  • Figure 2 shows the effect of Shanxidu on tumor volume.
  • Figure 3 shows the inhibitory effect of high-dose and low-dose Shanxi multitumor growth.
  • Subculture of cell lines of different cancer types including cancerous lung cancer, gastric cancer, liver cancer, rectal cancer, skin cancer, cervical cancer, prostate cancer, bladder cancer, breast cancer, blood cancer, pancreatic cancer, ovarian cancer, tongue cancer Osteosarcoma, kidney cancer, and the control group also used normal cells of the kidney (HEK293 (Kidney)) and lung epithelial cells BEAS-2B (Lung Epithelial) for testing.
  • HEK293 Kidney
  • BEAS-2B Lung Epithelial
  • the cells corresponding to different species should be counted with the corresponding culture solution (Table 1), and the number of cells should be counted, and the number of cells should be 2 ⁇ 10 6 cells.
  • the culture medium to which the cell strain was cultured was added to a volume of 10 ml, and the culture was continued for 2-3 days. The cells were then counted and dispensed into 96-well plates where the number of cells per well was fixed at 3,000 and the volume was 100 ul.
  • the original culture solution in the 96-well plate was aspirated, and a commercially available drug having a concentration of 10 ⁇ m and a volume of 100 ul was added to each well. After 72 hours, 100 ul of diluted WST-1 reagent was added to each well, and the dilution ratio was the culture solution and The volume ratio of the WST-1 stock solution is 9:1. Finally, the total volume of each well plate is 200 ul. Then, the 96-well plate is placed at 37 ° C for 30-90 minutes, using an elisa reader (ELISA) on the OD450. The absorbance values were detected and the survival rate of each cancer cell line was calculated.
  • ELISA elisa reader
  • the gastric cancer cell lines were AGS and MKN-45, respectively (Table 9), and the liver cancer cell lines were respectively For HepG2 and Hep3B cell lines (Table 10), the colorectal cancer cell lines were HCT116-wt and LoVo cell lines, respectively (Table 11), and the pancreatic cancer cell lines were AsPC and BxPC cell lines, respectively (Table 12).
  • the tongue cancer cell line was a SAS cell strain (Table 13), and the results of the cancer cell inhibition experiments were performed four times each, and the average value was calculated, and the results are shown below.
  • the osteosarctic cancer cell line is U2OS cell line (Table 14)
  • the skin cancer cell line is A375 and BCC two cell lines (Table 15), each doing 3 to 4 times. Cancer The results of the cytostatic experiments were calculated, and the average values were calculated, and the results are shown below.
  • the normal kidney cell line is HEK293 cell line (Table 16)
  • the normal lung epithelial cell line is BEAS-2B cell line (Table 17)
  • each doing 4 times of cancer The results of the cytostatic experiments were calculated, and the average values were calculated, and the results are shown below.
  • Table 18 Summary of the inhibition effect of Shan Xiduo on various cancer cells
  • mice Female BALB/cAnN.Cg-Foxn1nu/CrlNarl mice were used as samples (purchased from National Laboratory Animal Center), weighing 21 ⁇ 1g, subcutaneously injected into gastric cancer cells (AGS) and randomly divided into cages.
  • the test drugs were divided into three groups: Normal control group, low dose (100 mg/kg/day), high dose (200 mg/kg/day).
  • the tumor formed more than 100 mm 3
  • the drug was administered by intraperitoneal injection every day; the tumor size was measured twice a week, and the tumor volume measurement formula was as follows: (L ⁇ W2); L represents the longest diameter of the tumor; W represents the shortest diameter of the tumor, and its details
  • the data is shown in Table 19.

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Abstract

本发明提供山喜多（麦考酚酸吗乙酯）在制备抑制癌症的医药组合物中的用途。

Description

山喜多药物在用于制备抑制癌症的医药组合物中的用途 技术领域
本发明为山喜多药物的新适应症的应用，尤其该山喜多药物具有抑制多种癌症的用途。
背景技术
山喜多(CellCept,Mycophenolate mofetil、麦考酚酸吗乙酯、麦考酚酸莫酯)为一种免疫抑制剂。主成分为mycophenolic acid(MPA、麦考酚酸)的2-morpholinoethyl ester(霉酚酸酯)。MPA为具选择性、非竞争性及可逆的强力纤维核苷单磷酸去氢酶(IMPDH)抑制剂，因此能抑制鸟嘌呤核苷酸合成的重新(denovo)路径而不与DNA结合，MPA对淋巴球造成的细胞生长抑制(cytostatic)强过对其他细胞的作用。CellCept用于预防接受同种异体肾脏移植的病人器官排斥的发生，并可治疗难以控制的肾脏排斥，以及用于接受同种异体心脏或肝脏移植的病人预防器官排斥，皆相当有效。山喜多是早已被FDA所认可的用药，具有大量药物机转及人体研究成果资料。
由于临床应用上的明显差异，因此从来没有人认为山喜多具有抑制癌症细胞的可能性。
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。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。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。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。
因此，如何能让癌症药物的快速的可以被开发出来，并且尽可能的减少临床失败的机率，在癌症病学上是一个非常急迫又重要的课题。
发明内容
为解决上述的问题，本发明针对山喜多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山喜多药物对正常细胞没有或仅有微小的毒性，但山喜多在肿瘤细胞之间具有选择性的影响。
本发明提供一种山喜多药物在用于制备抑制癌症的医药组合物中用途，其中所述医药组合物包含有效剂量的山喜多药物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、乳癌及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、胰脏癌及舌癌中的一种或多种。
本发明一实施例中，其中所述山喜多药物的有效剂量浓度为20～500mg/kg/ 天。
附图说明
图1显示本发明山喜多用药应用于抑制癌细胞分析结果。
图2为山喜多可对肿瘤体积的效果。
图3为高剂量及低剂量山喜多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
Figure PCTCN2015092777-appb-000001
Figure PCTCN2015092777-appb-000002
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。
山喜多药对于各种癌细胞分析结果
山喜多对胸腔相关癌症细胞抑制效果的测试
本分析山喜多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、山喜多对肺癌相关癌症细胞抑制测试
  0524-10min 0526-10min 0529-10min 0531-10min 平均
A549 28.8 47.0 34.0 40.8 37.7
  1-10min 2-20min 3-20min 4-20min 平均
H1650 82.5 47.7 74.4 102.4 76.7
山喜多对腹腔相关癌症细胞抑制效果的测试
本分析山喜多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、山喜多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 57.4 39.1 38.0 38.8 43.3
  T24-1-30min T24-2-20min T24-3-20min T24-4-20min 平均
T24 45.1 32.5 41.1 48.0 41.6
表四、山喜多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 54.3 67.9 41.5 43.8 51.9
C-33A 45.4 46.0 44.2 50.1 46.4
表五、山喜多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 27.8 26.2 20.6 20.5 23.8
山喜多对内分泌相关癌症细胞抑制效果的测试
本分析山喜多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、山喜多对前列腺癌相关癌症细胞抑制测试
Figure PCTCN2015092777-appb-000003
表七、山喜多对乳癌相关癌症细胞抑制测试
Figure PCTCN2015092777-appb-000004
表八、山喜多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 63.7 64.8 74.2 75.7 69.6
  7-3-30min 7-4-30min 7-7-30min -4-30min 平均
TOV-21G 53.5 52.2 46.3 34.6 46.6
山喜多对消化道相关癌症细胞抑制效果的测试
分析山喜多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、山喜多对胃癌相关癌症细胞抑制测试
  0510-10min 0512-10min 0515-10min 0517-10min 平均
AGS 29.6 30.1 30.2 32.8 30.7
  0510-10min 0512-10min 0515-10min 0517-10min 平均
MKN-45 58.0 55.3 49.6 76.4 59.8
表十、山喜多对肝癌相关癌症细胞抑制测试
  0524-20min 0526-20min 0529-20min 0531-20min 平均
HepG2 48.1 75.0 75.8 75.1 68.5
  0612-20min 0614-20min 0616-20min 0619-20min 平均
Hep3B 86.3 76.4 66.2 63.3 73.1
表十一、山喜多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 35.1 38.8 41.8 38.9 38.7
  0616-10min 0619-10min 0621-10min 0623-10min 平均
LoVo 79.5 78.3 74.5 64.1 74.1
表十二、山喜多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 57.0 81.3 58.8 77.8 68.7
  3-7-3-30min 3-7-4-30min 3-7-7-30min 3-4-30min 平均
BxPC 40.8 84.0 54.5 67.9 61.8
表十三、山喜多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 47.7 45.1 46.4 42.4 45.4
山喜多对其他癌症细胞抑制效果的测试
分析山喜多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、山喜多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 58.9 54.4 59.7 54.1 56.8
表十五、山喜多对皮肤癌相关癌症细胞抑制测试
  0602-30min 0605-10min 0607-10min 0609-10min 平均
A375 28.7 28.2 22.9 22.7 25.6
  0602-30min 0605-10min 0607-10min 0609-10min 平均
BCC 43.5 33.6 40.7   39.3
对照组实验设计
山喜多对正常细胞抑制效果的测试
分析山喜多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、山喜多对正常肾脏细胞抑制测试
  平均
HEK293 86.9
表十七、山喜多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 23.6 27.6 27.6 33.2 28.0
针对山喜多对各种癌症细胞抑制效果汇整于表十八中，清楚看出山喜多对于多项癌症有明显的抑制效果。经过发明人实验结果，山喜多药物对于不同的癌症细胞有明显的抑制效果。(图一)
表十八、山喜多对各种癌症细胞抑制效果的汇整
癌症细胞 抑制效果
肺癌 57.20231111
膀胱癌 42.5
子宫颈癌 51.87
前列腺癌 50.69
乳癌 65.44
卵巢癌 58.1
胃癌 45.2
肝癌 70.79
大肠癌 56.38
胰脏癌 65.2
舌癌 45.41
骨肉癌 56.80
皮肤癌 32.47
肾脏癌 72.5
肾脏 86.91
肺部上皮细胞 28.01
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，皮下注射胃癌细胞(AGS)后随机分笼，测试药物分成三组:正常对照组、低剂量(100mg/kg/天)、高剂量(200mg/kg/天)。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，其详细数据如表19所示。
表十九、山喜多于动物实验对癌症对癌症的抑制效果
Figure PCTCN2015092777-appb-000005
Figure PCTCN2015092777-appb-000006
依据第2图的结果，低剂量与高剂量的山喜多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，因此表示山喜多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，高低剂量的山多喜可有效减缓肿瘤体积增长，并可同时减少肿瘤体积，其中以高剂量的山喜多具有较佳之效果。
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. 一种山喜多药物在用于制备抑制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山喜多(CellCept)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每日20mg/kg～500mg/kg。
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PCT/CN2015/092782 WO2016062291A1 (zh) 2014-10-24 2015-10-23 千忧解药物在用于制备治疗癌症的医药组合物中的用途
PCT/CN2015/092714 WO2016062275A1 (zh) 2014-10-24 2015-10-23 阿折地平在制备治疗癌症的医药组合物中的用途
PCT/CN2015/092653 WO2016062267A1 (zh) 2014-10-24 2015-10-23 奈必洛尔在制备抑制癌症的医药组合物中的用途
PCT/CN2015/092775 WO2016062285A1 (zh) 2014-10-24 2015-10-23 神经系统疾病用药在制备抗癌医药组合物中的应用
PCT/CN2015/092771 WO2016062283A1 (zh) 2014-10-24 2015-10-23 抗发炎用药物在制备抑制癌症的医药组合物中的应用
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PCT/CN2015/092702 WO2016062274A1 (zh) 2014-10-24 2015-10-23 消化系统疾病用药物在制备抑制癌症的医药组合物中的应用
PCT/CN2015/092666 WO2016062269A1 (zh) 2014-10-24 2015-10-23 阿那格雷在用于制备治疗癌症的医药组合物中的用途
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PCT/CN2015/092686 WO2016062271A1 (zh) 2014-10-24 2015-10-23 抗生素药物用于制备治疗癌症的医药组合物的用途
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