WO2016062265A1 - 莫诺苯宗药物在用于制备治疗癌症的医药组合物中的用途 - Google Patents
莫诺苯宗药物在用于制备治疗癌症的医药组合物中的用途 Download PDFInfo
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- WO2016062265A1 WO2016062265A1 PCT/CN2015/092617 CN2015092617W WO2016062265A1 WO 2016062265 A1 WO2016062265 A1 WO 2016062265A1 CN 2015092617 W CN2015092617 W CN 2015092617W WO 2016062265 A1 WO2016062265 A1 WO 2016062265A1
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- A61K38/00—Medicinal preparations containing peptides
Definitions
- the present invention is an application of a new indication for a monobenzone drug, in particular, the use of the monobenzone drug for inhibiting various cancers.
- Monobenzone is a topical decolorizing agent used to treat hyperpigmentation, such as various pigmentation, senile plaques, melanoma, etc., and the effect is obvious. It can break down the melanin in the skin, prevent the formation of melanin in the skin, restore the skin to a healthy color, without destroying the melanocytes, the toxicity is very light, usually made into an ointment or tincture. Monobenzone is a drug that has long been approved by the FDA and has a large number of drug transfer 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 successively develop drug resistance, which greatly reduces the effectiveness of drug use, and ultimately leads 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 monotil drugs, and achieves the goal of new use of old drugs.
- the experimental design results show that the Monobenzone drug has no or only minimal toxicity to normal cells, but Monobenzone has a selective effect on tumor cells.
- the invention provides a pharmaceutical composition for preparing cancer for treating monoclofen
- the use of the pharmaceutical composition comprising an effective amount of a monobenzone 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 monoxantine drug is 20 to 500 mg/kg/day.
- Fig. 1 shows the results of application of the monoxantine of the present invention for inhibiting cancer cell analysis.
- FIG. 2 shows the effect of monobenzone on tumor volume
- Figure 3 shows the inhibitory effect of high-dose and low-dose monobenzone on tumor 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 in 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 3000 cells 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 analysis of the inhibitory effect of monoxantine on the cells of the abdominal cavity-related cancer was mainly tested on two types of cells of the abdominal cavity-related cancer.
- the bladder cancer cell lines were TSGH and T24, respectively (Table 3), cervical cancer cells.
- the strains were HeLa cell line (Table 4), and kidney cancer was 786-O cell strain (Table 5).
- the results of cancer cell inhibition experiments were performed 4 times each, and the average value was calculated. The results are shown below.
- the analysis of the inhibitory effect of monoxantine on the cells of the abdominal cavity-related cancer was mainly tested against three types of endocrine-related cancer cells, which were PC-3 and LNCap, respectively (Table 6), breast cancer cells.
- the strains were MCF7 and MDA-MB-231, respectively (Table 7), and the ovarian cancer cell lines were NIH-OVCAR-3 cell line and TOV-21G (Table 8), respectively.
- the results are calculated and their average values are calculated, and the results are listed below.
- the gastric cancer cell lines are AGS and MKN-45, respectively (Table 9), liver cancer cells.
- the strains were HepG2 and Hep3B, respectively (Table 10)
- the colorectal cancer cell lines were HCT116-wt and LoVo cell lines (Table 11)
- 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 was U2OS cell line (Table 14)
- the skin cancer cell lines were A375 and BCC cell lines (Table 15)
- the normal kidney cell line is HEK293 cell line (Table 16)
- the normal fibroblast cell line is HFW cell line (Table 17)
- the normal lung epithelial cell line is HEK293 cell line (Table 18)
- the results of the cancer cell inhibition experiments were performed 4 times each, and the average value was calculated, and the results are shown below.
- 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.
- the test results are shown in Table 20.
- both low-dose and high-dose monobenzone have good inhibitory effects on tumors, and the weight of mice in each group did not decrease significantly during the experiment. Both the high and low doses can make the tested mice have good health without death during the treatment.
- high and low doses of monobenzone can effectively reduce tumor volume growth and simultaneously reduce tumor volume, with high doses of monobenzone having better effects.
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Abstract
本发明提供了莫诺苯宗在制备治疗癌症的医药组合物中的用途。
Description
本发明为莫诺苯宗药物的新适应症的应用,尤其该莫诺苯宗药物具有抑制多种癌症的用途。
莫诺苯宗(Monobenzone)是一种局部使用的脱色剂,用于治疗色素沉着过度,如各种色斑、老年斑、黑色素瘤等,效果很明显。它能分解皮肤中的黑色素,阻止皮肤中黑色素的生成,使皮肤恢复健康的色泽,而并不破坏黑色素细胞,毒性很轻,通常制成软膏或搽剂。莫诺苯宗是早已被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莫诺苯宗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,本发明针对莫诺苯宗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莫诺苯宗药物对正常细胞没有或仅有微小的毒性,但莫诺苯宗在肿瘤细胞之间具有选择性的影响。
本发明提供一种莫诺苯宗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包含有效剂量的莫诺苯宗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,其中所述莫诺苯宗药物的有效剂量浓度为20~500mg/kg/天。
图1显示本发明莫诺苯宗用药应用于抑制癌细胞分析结果。
图2显示莫诺苯宗用药可对肿瘤体积的效果
图3显示高剂量及低剂量莫诺苯宗对肿瘤生长的抑制效果
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,将2×106细胞数至入培养皿中,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个细胞,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。
莫诺苯宗药对于各种癌细胞分析结果
莫诺苯宗对胸腔相关癌症细胞抑制效果的测试
本分析莫诺苯宗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、莫诺苯宗对肺癌相关癌症细胞抑制测试
0524-10min | 0526-10min | 0529-10min | 0531-10min | 平均 | |
A549 | 80.8 | 111.1 | 80.8 | 100.9 | 93.4 |
1-10min | 2-20min | 3-20min | 4-20min | 平均 | |
H1650 | 90.1 | 66.6 | 86.7 | 101.8 | 86.3 |
莫诺苯宗对腹腔相关癌症细胞抑制效果的测试
本分析莫诺苯宗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、莫诺苯宗对膀胱癌相关癌症细胞抑制测试
表四、莫诺苯宗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 | 82.8 | 72.8 | 82.7 | 81.9 | 80.1 |
1 | 2 | 3 | 4 | 平均 | |
C-33A | 57.9 | 66.2 | 61.0 | 64.3 | 62.3 |
表五、莫诺苯宗对肾脏癌相关癌症细胞抑制测试
1 | 2 | 3 | 4 | 平均 | |
786-O | 39.8 | 32.4 | 21.2 | 23.4 | 29.2 |
莫诺苯宗对内分泌相关癌症细胞抑制效果的测试
本分析莫诺苯宗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、莫诺苯宗对前列腺癌相关癌症细胞抑制测试
表七、莫诺苯宗对乳癌相关癌症细胞抑制测试
表八、莫诺苯宗对卵巢癌相关癌症细胞抑制测试
莫诺苯宗对消化道相关癌症细胞抑制效果的测试
分析莫诺苯宗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、莫诺苯宗对胃癌相关癌症细胞抑制测试
表十、莫诺苯宗对肝癌相关癌症细胞抑制测试
0524-20min | 0526-20min | 0529-20min | 0531-20min | 平均 | |
HepG2 | 88.2 | 85.6 | 90.8 | 74.3 | 84.7 |
0612-20min | 0614-20min | 0616-20min | 0619-20min | 平均 | |
Hep3B | 110.6 | 116.2 | 96.8 | 100.6 | 106.1 |
表十一、莫诺苯宗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 | 91.12 | 103.66 | 99.94 | 85.3 | 95.0 |
0616-10min | 0619-10min | 0621-10min | 0623-10min | 平均 | |
LoVo | 82.29 | 73.94 | 94.7 | 81.3 | 83.0 |
表十二、莫诺苯宗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 | 88.7 | 116.5 | 81.7 | 93.7 | 95.1 |
3-7-3-30min | 3-7-4-30min | 3-7-7-30min | 3-4-30min | 平均 | |
BxPC | 57.4 | 100.0 | 70.7 | 86.3 | 78.6 |
表十三、莫诺苯宗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 | 97.29 | 59.36 | 89.56 | 115.2 | 90.3 |
莫诺苯宗对其他癌症细胞抑制效果的测试
分析莫诺苯宗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、莫诺苯宗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 | 69.3 | 66.7 | 73.0 | 81.7 | 72.7 |
表十五、莫诺苯宗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 | 56.7 | 64.8 | 77.3 | 106.8 | 76.4 |
0602-30min | 0605-10min | 0607-10min | 0609-10min | 平均 | |
BCC | 48.2 | 59.8 | 38.6 | 53.80 | 50.1 |
对照组实验设计
莫诺苯宗对正常细胞抑制效果的测试
分析莫诺苯宗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),正常纤维母细胞细胞株为HFW细胞株(表十七),正常肺部上皮细胞细胞株为BEAS-2B细胞株(表十八),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。
表十六、莫诺苯宗对正常肾脏细胞抑制测试
0602-30min | 0605-30min | 0607-30min | 0609-30min | 平均 | |
HEK293 | 82.7 | 99.43 | 97.88 | 91.84 | 93.0 |
表十七、莫诺苯宗对正常纤维母细胞抑制测试
0612-10min | 0614-10min | 0616-10min | 0619-10min | 平均 | |
HFW | 119.66 | 74.93 | 68.08 | 79.13 | 85.4 |
表十八、莫诺苯宗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 | 84.7 | 79.5 | 74.9 | 56.2 | 73.8 |
针对莫诺苯宗对各种癌症细胞抑制效果汇整于表十九中,清楚看
出莫诺苯宗对于多项癌症有明显的抑制效果。经过发明人实验结果,莫诺苯宗药物对于不同的癌症细胞有明显的抑制效果。(图一)
表十九、莫诺苯宗对各种癌症细胞抑制效果的汇整
癌症细胞 | 抑制效果 |
肺癌 | 89.85 |
膀胱癌 | 85.80 |
子宫颈癌 | 71.20 |
肾脏癌 | 29.20 |
前列腺癌 | 65.20 |
乳癌 | 87.90 |
卵巢癌 | 84.95 |
胃癌 | 82.80 |
肝癌 | 95.40 |
大肠癌 | 89.00 |
胰脏癌 | 86.85 |
舌癌 | 90.30 |
骨肉癌 | 72.70 |
皮肤癌 | 63.25 |
正常细胞 | 抑制效果 |
肾脏 | 93.00 |
肺部上皮细胞 | 73.80 |
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。试验结果如表二十所示。
依据第2图的结果,低剂量与高剂量的莫诺苯宗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,因此表示莫诺苯宗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,高低剂量的莫诺苯宗可有效减缓肿瘤体积增长,并可同时减少肿瘤体积,其中以高剂量的莫诺苯宗具有较佳的效果。
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)
- 一种莫诺苯宗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莫诺苯宗(Monobenzone)及药学上可接受的盐类所组成。
- 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。
- 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、皮肤癌相关癌症中的一种或多种。
- 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。
- 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、子宫颈癌、肾脏癌中的一种或多种。
- 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。
- 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。
- 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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