Abstract:
Provided in the present invention are uses of duloxetine hydrochloride in preparing a pharmaceutical composition for treatment of cancer.

Description:
CROSS-REFERENCE TO RELATED APPLICATION 
       [0001]    This application is a bypass continuation of International Application No. PCT/CN2015/092782, filed on Oct. 23, 2015, which claims priority of U.S. Provisional Application No. 62/068,298, filed on Oct. 24, 2014, the entire content of each of which is incorporated herein by reference. 
     
    
     FIELD 
       [0002]    The disclosure relates to the application of a duloxetine HCl medicament in a new indication, and more particularly to use of the duloxetine HCl medicament in inhibiting various cancers. 
       BACKGROUND 
       [0003]    Cancer has been the first leading cause of death globally for a long time and the patients suffering from cancer are increasing year by year. Therefore, the treatment of cancer is becoming an important topic. Cancer treatment can be divided into surgical treatment, radiation therapy, chemotherapy and targeted therapy. 
         [0004]    Generally, the goal of cancer drug treatment, either chemotherapy or targeted therapy, is mainly to suppress the metastasis and expansion of cancer cells by inhibiting their replication and division. 
         [0005]    Cancer drug design also aims at the development of a drug molecule having high specificity or at the design of a targeted antibody. According to statistical data, only about five drugs per ten thousand new drugs are able to access phase I clinical trials. 
         [0006]    In addition to the problem of large-scale production of the drugs, other issues that still need to be overcome are drug safety, patient selection and testing dosage, etc. Even if the drug has been approved by the FDA and has been marketed, it is usually the case that a patient&#39; s response to the drug is not favorable. Furthermore, many cancer cells may start becoming resistant to the drug, which causes the significant reduction of efficacy of the drug, thereby resulting in failure of the cancer treatment. Therefore, there is a certain level of difficulty with regard to drug development. 
         [0007]    Duloxetine hydrochloride (Duloxetine HCl) is a selective serotonin and norepinephrine reuptake inhibitor (SSNRI) for oral administration. The anti-depression mechanism of duloxetine in human bodies is unknown, but it is assumed that such mechanism is associated with the serotonergic and noradrenergic activities of duloxetine acting upon the central nervous system. Pre-clinical tests have shown that duloxetine can effectively block the reuptake of serotonin and norepinephrine, but merely slightly block the reuptake of dopamine. In vitro tests have shown that duloxetine has no significant affinity to dopaminergic, adrenergic, cholinergic or histaminergic receptors. Duloxetine does not inhibit monoamine oxidase. Duloxetine undergoes extensive metabolism, but it has not been found that the major circulating metabolites have significant pharmacologic activity of duloxetine. Duloxetine HCl has been approved by the FDA and there is a large amount of human research result data regarding Duloxetine HCl. 
         [0008]    Due to the significant differences in clinical applications, duloxetine HCl has never been considered to have potential to inhibit cancer cells. 
       SUMMARY 
       [0009]    In order to resolve the aforementioned problems, the present invention is directed to the development of the new indication of a duloxetine HCl medicament, thereby achieving the purpose to use the known drug for a new indication. 
         [0010]    It is shown via the designed experimental results that the duloxetine HCl medicament has little or no cytotoxicity to the normal cells, and duloxetine HCl exhibits selective effect between normal cells and tumor cells. 
         [0011]    The invention provides use of a duloxetine HCl medicament in the manufacture of a pharmaceutical composition for treating cancer, in which the pharmaceutical composition is selected from an effective dosage of duloxetine and a pharmaceutically acceptable salt thereof. 
         [0012]    In an embodiment of the present invention, the aforesaid cancer may be one or more cancers selected from thorax-related cancer, peritoneal cavity-related cancer, endocrine-related cancer and digestive tract-related cancer. 
         [0013]    In an embodiment of the present invention, the aforesaid cancer may be one or more cancers selected from osteosarcoma-related cancer, skin-related cancer and leukemia-related cancer. 
         [0014]    In an embodiment of the present invention, the aforesaid thorax-related cancer may be lung cancer. 
         [0015]    In an embodiment of the present invention, the aforesaid peritoneal cavity-related cancer may be selected from bladder cancer and/or cervical cancer. 
         [0016]    In an embodiment of the present invention, the aforesaid endocrine-related cancer may be one or more cancers selected from prostate cancer, breast cancer and ovarian cancer. 
         [0017]    In an embodiment of the present invention, the aforesaid digestive tract-related cancer may be one or more cancers selected from gastric cancer, liver cancer, colon cancer, pancreatic cancer and lingual cancer. 
         [0018]    In an embodiment of the present invention, the effective dosage of the duloxetine HCl medicament may range from 20 mg/kg/day to 500 mg/kg/day. 
     
    
     
       BRIEF DESCRIPTION OF THE DRAWINGS 
         [0019]      FIG. 1  shows the analysis results of duloxetine HCl of the present invention for use in inhibiting cancer cells; 
           [0020]      FIG. 2  shows the effect of duloxetine HCl on tumor volume; and 
           [0021]      FIG. 3  shows the inhibitory effect of duloxetine HCl in high dosage and low dosage on tumor growth. 
       
    
    
     DETAILED DESCRIPTION 
     Development of Various Cancer Cell Lines 
       [0022]    Different types of cancer cell lines were sub-cultured. These cancer cell types include lung cancer, gastric cancer, liver cancer, colorectal cancer, skin cancer, cervical cancer, prostate cancer, bladder cancer, breast cancer, leukemia, pancreatic cancer, ovarian cancer, lingual cancer, osteosarcoma and renal cancer. The normal cells of the kidney (HEK2 93) and lung epithelium BEAS -2 B serve as the control group to conduct the test (Table 1). 
         [0023]    After cultivating each cell in the culture medium (due to different cell characteristics, each of the cancer cell lines was cultivated in the corresponding culture medium (Table 1)), the cell numbers were counted. These cells were seeded at an amount of 2×10 6  cells, and then added with the culture medium for cultivating the cells to a volume of 10 mL, followed by cultivation for 2 to 3 days. After counting the cell number, these cells were respectively plated in a 96-well plate, in which each well has a fixed cell number of 3000 and a volume of 100 μL. 
         [0000]    
       
         
               
             
               
               
               
               
             
               
               
               
               
             
           
               
                 TABLE 1 
               
             
             
               
                   
               
               
                 Test cancer cell lines and the culture medium 
               
               
                 for cultivating the same 
               
             
          
           
               
                   
                   
                   
                 Culture 
               
               
                 No 
                 Cancer 
                 Cancer cell type 
                 medium 
               
               
                   
               
             
          
           
               
                 1 
                 Lung 
                 H1650 (lung adenocarcinoma) 
                 RPMI-1640 
               
               
                   
                 cancer 
                 A549 (lung adenocarcinoma) 
                 DMEM 
               
               
                 2 
                 Gastric 
                 AGS (gastric adenocarcinoma) 
                 RPMI-1640 
               
               
                   
                 cancer 
                 MKN-45 (gastric adenocarcinoma) 
                 RPMI-1640 
               
               
                 3 
                 Liver 
                 HepG2 (hepatocellular carcinoma) 
                 DMEM 
               
               
                   
                 cancer 
                 Hep3B (hepatocellular carcinoma) 
                 DMEM 
               
               
                 4 
                 ColorRectal 
                 HCT116 (p53+) (colorectal carcinoma) 
                 DMEM 
               
               
                   
                 cancer 
                 LoVo (colorectal adenocarcinoma) 
                 DMEM 
               
               
                 5 
                 Skin 
                 A375 (amelanotic melanoma) 
                 DMEM 
               
               
                   
                 cancer 
                 BCC (basal cell carcinoma) 
                 DMEM 
               
               
                 6 
                 Cervical 
                 HeLa (cervix adenocarcinoma) 
                 DMEM 
               
               
                   
                 cancer 
                 C-33A (cervical carcinoma) BCRC 60554 
                 MEM 
               
               
                 7 
                 Prostate 
                 PC3 (p53−) (prostate adenocarcinoma) 
                 DMEM 
               
               
                   
                 cancer 
                 LNCaP clone FGC (LNCap.FGC) 
                 RPMI-1640 
               
               
                 8 
                 Bladder 
                 TSGH (urinary bladder carcinoma) 
                 RPMI-1640 
               
               
                   
                 cancer 
                 T24 
                 RPMI-1640 
               
               
                 9 
                 Breast 
                 MCF7 (mammary gland, adenocarcinoma) 
                 DMEM 
               
               
                   
                 cancer 
                 MDA-MB-231 (mammary gland, 
                 DMEM 
               
               
                   
                   
                 adenocarcinoma) 
               
               
                 10 
                 Pancreatic 
                 BxPC-3 
                 RPMI-1640 
               
               
                   
                 cancer 
                 AsPC-1 
                 RPMI-1640 
               
               
                 11 
                 Ovarian 
                 NIH:OVCAR-3 
                 RPMI-1640 
               
               
                   
                 cancer 
                 TOV-21G 
                 RPMI-1640 
               
               
                 12 
                 Lingual 
                 SAS (tongue squamouscell carcinoma) 
                 DMEM 
               
               
                   
                 cancer 
               
               
                 13 
                 Osteosarcoma 
                 U-2OS 
                 DMEM 
               
               
                 14 
                 renal 
                 786-O (renal adenocarcinoma) BCRC 60243 
                 RPMI-1640 
               
               
                   
                 cancer 
               
               
                 15 
                 Normal cell 
                 HEK293 (kidney) 
                 DMEM 
               
               
                   
                 Kidney 
                 BEAS-2B (lung epithelial) 
                 RPMI-1640 
               
               
                   
                 Lung 
               
               
                   
                 epithelium 
               
               
                   
               
             
          
         
       
     
       Cell Survival Analysis 
       [0024]    After removing the culture medium in the 96-well plate, each well was added with 100 μL of 10 μM commercial drug and then placed for 72 hours. Thereafter, each well was further added with 100 μL of a diluted WST-1 solution (the dilution ratio for the culture medium: WST-1 stock solution (wt/wt) is 9:1). The 96-well plate (each well having a final volume of 200 μL) was placed at 37° C. for 30 to 90 minutes. Absorbance was detected at OD 450  with an elisa reader (enzyme-linked immunosorbent detector), and the survival rate of each of the cancer cell lines was calculated. A lower survival rate (%) indicates that the drug has a stronger inhibitory effect against the test cancer cell. 
       Analysis Result of Duloxetine HCl Against Various Cancer Cells 
     The Test on Inhibitory Effect of Duloxetine HCl Against Thorax-Related Cancer Cells 
       [0025]    This analysis entitled “The test on inhibitory effect of duloxetine HCl against thorax-related cancer cells” was mainly directed to two types of lung cancer cells, i.e. A549 and H1650 cancer cell lines, for conducting the test. Each cancer cell inhibitory experiment was repeated for 4 times, followed by calculation of the mean values thereof, which were shown in the following table (Table 2). 
         [0000]    
       
         
               
             
               
               
               
               
               
               
             
           
               
                 TABLE 2 
               
               
                   
               
               
                 The inhibitory test of duloxetine HCl against 
               
               
                 lung cancer-related cancer cells 
               
               
                   
               
             
             
               
                   
               
             
          
           
               
                   
                   
                   
                   
                   
                 Mean 
               
               
                   
                   
                   
                   
                 0531-10 
                 survival 
               
               
                   
                 0524-10 min 
                 0526-10 min 
                 0529-10 min 
                 min 
                 rate (%) 
               
               
                   
               
               
                 A549 
                 32.8 
                 53.7 
                 33.2 
                 30.1 
                 37.5 
               
               
                   
               
               
                   
                 1-10 min 
                 2-20 min 
                 3-20 min 
                 4-20 min 
                 Mean 
               
               
                   
               
               
                 H1650 
                 35.2 
                 36.9 
                 33.6 
                 39.4 
                 36.3 
               
               
                   
               
             
          
         
       
     
       The Test on Inhibitory Effect of Duloxetine HCl Against Peritoneal Cavity-Related Cancer Cells 
       [0026]    This analysis entitled “The test on inhibitory effect of duloxetine HCl against peritoneal cavity-related cancer cells” was mainly directed to three types of peritoneal cavity-related cancers for conducting the test. Bladder cancer cell lines were respectively TSGH and T24 cell lines (Table 3). Cervical cancer cell lines were respectively HeLa and C-33A cell lines (Table 4). Renal cancer cell line was 786-O cell line (Table 5). Each cancer cell inhibitory experiment was repeated for 4 times, followed by calculation of the mean values thereof, which were shown in the following tables. 
         [0000]    
       
         
               
             
               
               
               
               
               
               
             
           
               
                 TABLE 3 
               
               
                   
               
               
                 The inhibitory test of duloxetine HCl against 
               
               
                 bladder cancer-related cancer cells 
               
               
                   
               
             
             
               
                   
               
             
          
           
               
                   
                   
                   
                   
                   
                 Mean 
               
               
                   
                   
                   
                   
                 0517-10 
                 survival 
               
               
                   
                 0510-10 min 
                 0512-10 min 
                 0515-10 min 
                 min 
                 rate (%) 
               
               
                   
               
               
                 TSGH 
                 36.1 
                 59.0 
                 61.2 
                 40.5 
                 49.2 
               
               
                   
               
               
                   
                 T24- 
                 T24- 
                 T24- 
                 T24- 
               
               
                   
                 1-30 min 
                 2-20 min 
                 3-20 min 
                 4-20 min 
                 Mean 
               
               
                   
               
               
                 T24 
                 81.7 
                 90.6 
                 71.4 
                 97.2 
                 85.2 
               
               
                   
               
             
          
         
       
     
         [0000]    
       
         
               
             
               
               
               
               
               
               
             
               
               
               
               
               
               
             
           
               
                 TABLE 4 
               
             
             
               
                   
               
               
                 The inhibitory test of duloxetine HCl against 
               
               
                 cervical cancer-related cancer cells 
               
             
          
           
               
                   
                   
                   
                   
                   
                 Mean 
               
               
                   
                 0524-10 
                 0526-10 
                 0529-10 
                 0531-10 
                 survival 
               
               
                   
                 min 
                 min 
                 min 
                 min 
                 rate (%) 
               
               
                   
                   
               
             
          
           
               
                 HeLa 
                 27.7 
                 29.9 
                 25.4 
                 52.1 
                 33.8 
               
               
                 C-33A 
                 34.3 
                 52.4 
                 33.7 
                 42.4 
                 40.7 
               
               
                   
               
             
          
         
       
     
         [0000]    
       
         
               
             
               
               
               
               
               
               
             
               
               
               
               
               
               
             
           
               
                 TABLE 5 
               
             
             
               
                   
               
               
                 The inhibitory test of duloxetine HCl against 
               
               
                 renal cancer-related cancer cells 
               
             
          
           
               
                   
                   
                   
                   
                   
                 Mean 
               
               
                   
                 0524-10 
                 0526-10 
                 0529-10 
                 0531-10 
                 survival 
               
               
                   
                 min 
                 min 
                 min 
                 min 
                 rate (%) 
               
               
                   
                   
               
             
          
           
               
                 786-O 
                 60.4 
                 89.7 
                 49.0 
                 70.1 
                 67.3 
               
               
                   
               
             
          
         
       
     
       The Test on Inhibitory Effect of Duloxetine HCl Against Endocrine-Related Cancer Cells 
       [0027]    This analysis entitled “The test on inhibitory effect of duloxetine HCl against endocrine-related cancer cells” was mainly directed to three types of endocrine-related cancers for conducting the test. Prostate cancer cell lines were respectively PC-3 and LNCap cell lines (Table 6). Breast cancer cell lines were respectively MCF-7 and MDA-MB-231 cell lines (Table 7). Ovarian cancer cell lines were respectively NIH-OVCAR-3 and TOV-21G cell lines (Table 8). Each cancer cell inhibitory experiment was repeated for 4 times, followed by calculation of the mean values thereof, which were shown in the following tables. 
         [0000]    
       
         
               
             
               
               
               
               
               
               
             
               
               
               
             
           
               
                 TABLE 6 
               
               
                   
               
               
                 The inhibitory test of duloxetine HCl against 
               
               
                 prostate cancer-related cancer cells 
               
               
                   
               
             
             
               
                   
               
             
          
           
               
                   
                   
                   
                   
                   
                 Mean 
               
               
                   
                 PC-3-0524- 
                 PC-3-0526- 
                 PC-3-0529- 
                 PC-3-0531- 
                 survival 
               
               
                   
                 10 min 
                 10 min 
                 10 min 
                 10 min 
                 rate (%) 
               
               
                   
               
               
                 PC-3 
                 45.0 
                 70.9 
                 41.4 
                 25.1 
                 45.6 
               
               
                   
               
             
          
           
               
                   
                   
                 Mean 
               
               
                   
                   
               
               
                   
                 LNCap 
                 72.8 
               
               
                   
                   
               
             
          
         
       
     
         [0000]    
       
         
               
             
               
               
               
               
               
               
             
               
               
               
               
               
             
           
               
                 TABLE 7 
               
               
                   
               
               
                 The inhibitory test of duloxetine HCl against 
               
               
                 breast cancer-related cancer cells 
               
               
                   
               
             
             
               
                   
               
             
          
           
               
                   
                   
                   
                   
                   
                 Mean 
               
               
                   
                 0612- 
                   
                   
                   
                 survival 
               
               
                   
                 10 min 
                 0614-10 min 
                 0616-10 min 
                 0619-10 min 
                 rate (%) 
               
               
                   
               
               
                 MCF-7 
                 29.0 
                 37.3 
                 30.0 
                 28.5 
                 31.2 
               
               
                   
               
             
          
           
               
                   
                 0612-10 min 
                 0614-10 min 
                 0616-10 min 
                 Mean 
               
               
                   
               
               
                 MDA-MB-231 
                 53.4 
                 70.2 
                 74.3 
                 65.9 
               
               
                   
               
             
          
         
       
     
         [0000]    
       
         
               
             
               
               
               
               
               
               
             
           
               
                 TABLE 8 
               
               
                   
               
               
                 The inhibitory test of duloxetine HCl against 
               
               
                 ovarian cancer-related cancer cells 
               
               
                   
               
             
             
               
                   
               
             
          
           
               
                   
                   
                   
                   
                   
                 Mean 
               
               
                   
                 7-3-30 
                 7-4-30 
                 7-7-30 
                   
                 survival 
               
               
                   
                 min 
                 min 
                 min 
                 -4-30 min 
                 rate (%) 
               
               
                   
               
               
                 NIH-OVCAR-3 
                 64.8 
                 74.6 
                 78.5 
                 55.0 
                 68.2 
               
               
                   
               
               
                   
                 7-3-30 
                 7-4-30 
                 7-7-30 
               
               
                   
                 min 
                 min 
                 min 
                 -4-30 min 
                 Mean 
               
               
                   
               
               
                 TOV-21G 
                 30.9 
                 41.9 
                 34.7 
                 31.9 
                 34.9 
               
               
                   
               
             
          
         
       
     
       The Test on Inhibitory Effect of Duloxetine HCl Against Digestive Tract-Related Cancer Cells 
       [0028]    This analysis entitled “The test on inhibitory effect of duloxetine HCl against digestive tract-related cancer cells” was mainly directed to five types of digestive tract-related cancers for conducting the test. Gastric cancer cell lines were respectively AGS and MKN-45 cell lines (Table 9). Liver cancer cell lines were respectively HepG2 and Hep3B cell lines (Table 10). Colorectal cancer cell lines were respectively HCT116-wt and LoVo cell lines (Table 11). Pancreatic cancer cell lines were respectively AsPC-1 and BxPC-3 cell lines (Table 12). Lingual cancer cell line was SAS cell line (Table 13). Each cancer cell inhibitory experiment was repeated for 4 times, followed by calculation of the mean values thereof, which were shown in the following tables. 
         [0000]    
       
         
               
             
               
               
               
               
               
               
             
           
               
                 TABLE 9 
               
               
                   
               
               
                 The inhibitory test of duloxetine HCl against 
               
               
                 gastric cancer-related cancer cells 
               
               
                   
               
             
             
               
                   
               
             
          
           
               
                   
                   
                   
                   
                   
                 Mean 
               
               
                   
                 0510- 
                   
                   
                   
                 survival 
               
               
                   
                 10 min 
                 0512-10 min 
                 0515-10 min 
                 0517-10 min 
                 rate (%) 
               
               
                   
               
               
                 AGS 
                 25.0 
                 15.9 
                 37.3 
                 15.8 
                 23.5 
               
               
                   
               
               
                   
                 0510- 
               
               
                   
                 10 min 
                 0512-10 min 
                 0515-10 min 
                 0517-10 min 
                 Mean 
               
               
                   
               
               
                 MKN-45 
                 59.5 
                 75.3 
                 46.0 
                 57.9 
                 59.7 
               
               
                   
               
             
          
         
       
     
         [0000]    
       
         
               
             
               
               
               
               
               
               
             
           
               
                 TABLE 10 
               
               
                   
               
               
                 The inhibitory test of duloxetine HCl against 
               
               
                 liver cancer-related cancer cells 
               
               
                   
               
             
             
               
                   
               
             
          
           
               
                   
                   
                   
                   
                   
                 Mean 
               
               
                   
                 0524- 
                   
                   
                   
                 survival 
               
               
                   
                 20 min 
                 0526-20 min 
                 0529-20 min 
                 0531-20 min 
                 rate (%) 
               
               
                   
               
               
                 HepG2 
                 14.8 
                 44.7 
                 37.7 
                 38.9 
                 34.0 
               
               
                   
               
               
                   
                 0612- 
               
               
                   
                 20 min 
                 0614-20 min 
                 0616-20 min 
                 0619-20 min 
                 Mean 
               
               
                   
               
               
                 Hep3B 
                 41.4 
                 53.3 
                 36.2 
                 60.6 
                 47.9 
               
               
                   
               
             
          
         
       
     
         [0000]    
       
         
               
             
               
               
               
               
               
               
             
           
               
                 TABLE 11 
               
               
                   
               
               
                 The inhibitory test of duloxetine HCl against 
               
               
                 colorectal cancer-related cancer cells 
               
               
                   
               
             
             
               
                   
               
             
          
           
               
                   
                   
                   
                   
                   
                 Mean 
               
               
                   
                 0602-30 
                 0605-10 
                 0607-10 
                 0609-10 
                 survival 
               
               
                   
                 min 
                 min 
                 min 
                 min 
                 rate (%) 
               
               
                   
               
               
                 HCT116-wt 
                 52.1 
                 108.8 
                 127.4 
                 20.3 
                 77.1 
               
               
                   
               
               
                   
                   
                   
                   
                   
                 Mean 
               
               
                   
                 0616-10 
                 0619-10 
                 0621-10 
                 0623-10 
                 survival 
               
               
                   
                 min 
                 min 
                 min 
                 min 
                 rate (%) 
               
               
                   
               
               
                 LoVo 
                 44.8 
                 63.4 
                 49.9 
                 31.0 
                 47.3 
               
               
                   
               
             
          
         
       
     
         [0000]    
       
         
               
             
               
               
               
               
               
               
             
           
               
                 TABLE 12 
               
               
                   
               
               
                 The inhibitory test of duloxetine HCl against 
               
               
                 pancreatic cancer-related cancer cells 
               
               
                   
               
             
             
               
                   
               
             
          
           
               
                   
                   
                   
                   
                   
                 Mean 
               
               
                   
                 1-7-3-30 
                 1-7-4-30 
                 1-7-7-30 
                   
                 survival 
               
               
                   
                 min 
                 min 
                 min 
                 1-4-30 min 
                 rate (%) 
               
               
                   
               
               
                 AsPC-1 
                 75.7 
                 78.0 
                 26.0 
                 61.7 
                 60.3 
               
               
                   
               
               
                   
                   
                   
                   
                   
                 Mean 
               
               
                   
                 3-7-3-30 
                 3-7-4-30 
                 3-7-7-30 
                   
                 survival 
               
               
                   
                 min 
                 min 
                 min 
                 3-4-30 min 
                 rate (%) 
               
               
                   
               
               
                 BxPC-3 
                 62.1 
                 58.8 
                 22.7 
                 72.4 
                 54.0 
               
               
                   
               
             
          
         
       
     
         [0000]    
       
         
               
             
               
               
               
               
               
               
             
               
               
               
               
               
               
             
           
               
                 TABLE 13 
               
             
             
               
                   
               
               
                 The inhibitory test of duloxetine HCl against 
               
               
                 lingual cancer-related cancer cells 
               
             
          
           
               
                   
                   
                   
                   
                   
                 Mean 
               
               
                   
                   
                   
                   
                   
                 survival 
               
               
                   
                 6-26-10 min 
                 6-28-10 min 
                 6-30-10 min 
                 7-3-10 min 
                 rate (%) 
               
               
                   
                   
               
             
          
           
               
                 SAS 
                 45.7 
                 88.9 
                 96.2 
                 98.9 
                 82.4 
               
               
                   
               
             
          
         
       
     
       The Test on Inhibitory Effect of Duloxetine HCl Against Other Cancer Cells 
       [0029]    The analysis of duloxetine HCl against other types of cancers was performed. Osteosarcoma cell line was U2OS cell line (Table 14). Skin cancer cell lines were respectively A375 and BCC cell lines (Table 15). Each cancer cell inhibitory experiment was repeated for 3 or 4 times, followed by calculation of the mean values thereof, which were shown in the following tables. 
         [0000]    
       
         
               
             
               
               
               
               
               
               
             
               
               
               
               
               
               
             
           
               
                 TABLE 14 
               
             
             
               
                   
               
               
                 The inhibitory test of duloxetine HCl against 
               
               
                 osteosarcoma-related cancer cells 
               
             
          
           
               
                   
                   
                   
                   
                   
                 Mean 
               
               
                   
                 6- 
                   
                   
                   
                 survival 
               
               
                   
                 26-10 min 
                 6-28-10 min 
                 6-30-10 min 
                 7-3-10 min 
                 rate (%) 
               
               
                   
                   
               
             
          
           
               
                 U2OS 
                 26.6 
                 21.5 
                 56.6 
                 35.6 
                 35.1 
               
               
                   
               
             
          
         
       
     
         [0000]    
       
         
               
             
               
               
               
               
               
               
             
               
               
               
               
               
               
             
           
               
                 TABLE 15 
               
               
                   
               
               
                 The inhibitory test of duloxetine HCl against 
               
               
                 skin cancer-related cancer cells 
               
               
                   
               
             
             
               
                   
               
             
          
           
               
                   
                   
                   
                   
                   
                 Mean 
               
               
                   
                   
                   
                   
                   
                 survival 
               
               
                   
                 0602-30 min 
                 0605-10 min 
                 0607-10 min 
                 0609-10 min 
                 rate (%) 
               
               
                   
               
               
                 A375 
                 25.7 
                 16.2 
                 17.1 
                 67.0 
                 31.5 
               
               
                   
               
             
          
           
               
                   
                   
                 0602-30 min 
                 0605-10 min 
                 0607-10 min 
                 Mean 
               
               
                   
                   
               
               
                   
                 BCC 
                 43.9 
                 31.3 
                 69.0 
                 48.1 
               
               
                   
                   
               
             
          
         
       
     
       Experimental Design for Control Group 
     The Test on Inhibitory Effect of Duloxetine HCl Against Normal Cells 
       [0030]    The analysis of duloxetine HCl against many types of normal cells was performed. Normal kidney cell line was HEK293 cell line (Table 16). Normal lung epithelial cell line was BEAS-2B cell line (Table 17). Each cell inhibitory experiment was repeated for 4 times, followed by calculation of the mean values thereof, which were shown in the following tables. 
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                 TABLE 16 
               
             
             
               
                   
               
               
                 The inhibitory test of duloxetine HCl against 
               
               
                 normal kidney cells 
               
             
          
           
               
                   
                 Mean survival rate (%) 
               
               
                   
                   
               
             
          
           
               
                   
                 HEK 293 
                 62.4 
               
               
                   
                   
               
             
          
         
       
     
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                 TABLE 17 
               
             
             
               
                   
               
               
                 The inhibitory test of duloxetine HCl against 
               
               
                 normal lung epithelial cells 
               
             
          
           
               
                   
                   
                   
                   
                   
                 Mean 
               
               
                   
                 0510- 
                   
                   
                   
                 survival 
               
               
                   
                 10 min 
                 0512-10 min 
                 0515-10 min 
                 0517-10 min 
                 rate (%) 
               
               
                   
                   
               
             
          
           
               
                 BEAS-2B 
                 59.5 
                 76.3 
                 74.7 
                 79.6 
                 72.5 
               
               
                   
               
             
          
         
       
     
         [0031]    The inhibitory effect of duloxetine HCl against various cancer cells was tabulated in Table 18. It can be clearly seen that duloxetine HCl exhibits significant inhibitory effect against various cancers. It is noted from the inventors&#39; experimental results that duloxetine HCl may have significant inhibitory effects against various cancer cells (see  FIG. 1 ). 
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                 TABLE 18 
               
             
             
               
                   
               
               
                 Summary of inhibitory test of duloxetine HCl 
               
               
                 against various cancer cells 
               
             
          
           
               
                   
                 Cancer cells 
                 Survival rate (%) 
               
               
                   
                   
               
             
          
           
               
                   
                 Lung cancer 
                 36.8743253 
               
               
                   
                 Bladder cancer 
                 67.2 
               
               
                   
                 Cervical cancer 
                 36.76 
               
               
                   
                 Prostate cancer 
                 59.23 
               
               
                   
                 Breast cancer 
                 48.57 
               
               
                   
                 Ovarian cancer 
                 51.5 
               
               
                   
                 Gastric cancer 
                 41.6 
               
               
                   
                 Liver cancer 
                 40.95 
               
               
                   
                 Colorectal cancer 
                 62.21 
               
               
                   
                 Pancreatic cancer 
                 57.2 
               
               
                   
                 Lingual cancer 
                 82.44 
               
               
                   
                 Osteosarcoma 
                 35.09 
               
               
                   
                 Skin cancer 
                 39.79 
               
               
                   
                 Renal cancer 
                 67.3 
               
               
                   
                 Kidney 
                 62.44 
               
               
                   
                 Lung epithelial cells 
                 72.50 
               
               
                   
                   
               
             
          
         
       
     
       Animal Experimental Analysis 
       [0032]    This experiment was performed with female BALB/cAnN.Cg-Foxn1nu/CrlNarl mice with a weight of around 21±1 gas samples. After subcutaneous injection of liver cancer cells (HepG2), the mice were randomly divided into three groups based on the test drug, i.e., normal control group, low dosage group (100 mg/kg/day) and high dosage group (200 mg/kg/day). After the tumor was grown to be over 100 mm 3 , the mice were treated with the drug via intraperitoneal injection daily. The tumor size was measured two times per week. The formula for determining the tumor size was as follows: (L×W 2 )/2, in which L represents the longest diameter of the tumor and W represents the shortest diameter of the tumor. 
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                 TABLE 19 
               
               
                   
               
               
                 The inhibitory test of duloxetine HCl against cancers in animal experiments 
               
               
                   
               
             
             
               
                   
               
             
          
           
               
                   
                 Control group 
                   
               
             
          
           
               
                   
                 Increased 
                 Low dosage (100 mg/kg/day) 
               
             
          
           
               
                   
                 Weight 
                 L 
                 W 
                 Volume 
                 tumor volume 
                 Weight 
                 L 
                 W 
               
               
                   
                 (g) 
                 (mm) 
                 (mm) 
                 (mm 3 ) 
                 (mm 3 ) 
                 (g) 
                 (mm) 
                 (mm) 
               
               
                   
                   
               
             
          
           
               
                 First measurement 
               
             
          
           
               
                 A 
                 18.5 
                 7 
                 7 
                 171.5 
                 171.5 
                 19 
                 7 
                 5 
               
               
                 B 
                 22 
                 8 
                 6 
                 144 
                 144 
                 19 
                 8 
                 6 
               
               
                 C 
                 20.5 
                 9 
                 8 
                 288 
                 288 
                 18 
                 6 
                 6 
               
               
                 Mean 
                   
                   
                   
                 201.1667 
                 201.1667 
               
             
          
           
               
                 Second measurement 
               
             
          
           
               
                 A 
                 22 
                 7 
                 6 
                 126 
                 −45.5 
                 19 
                 6 
                 5 
               
               
                 B 
                 20 
                 8 
                 7 
                 196 
                 52 
                 21 
                 7 
                 6 
               
               
                 C 
                 20 
                 9 
                 7 
                 220.5 
                 −67.5 
                 20 
                 7 
                 6 
               
               
                 Mean 
                   
                   
                   
                 180.8333 
                 −20.3333 
               
             
          
           
               
                 Third measurement 
               
             
          
           
               
                 A 
                 23 
                 9 
                 6 
                 162 
                 36 
                 20.5 
                 7 
                 6 
               
               
                 B 
                 20 
                 10 
                 8 
                 320 
                 124 
                 23.5 
                 7 
                 4 
               
               
                 C 
                 21 
                 11 
                 7 
                 269.5 
                 49 
                 19 
                 7 
                 5 
               
               
                 Mean 
                   
                   
                   
                 250.5 
                 69.66667 
               
             
          
           
               
                 Fourth measurement 
               
             
          
           
               
                 A 
                 23 
                 11 
                 7 
                 269.5 
                 107.5 
                 22 
                 6 
                 6 
               
               
                 B 
                 22 
                 10 
                 6 
                 180 
                 −140 
                 22 
                 4 
                 4 
               
               
                 C 
                 23 
                 11 
                 8 
                 352 
                 82.5 
                 20 
                 6 
                 5 
               
               
                 Mean 
                   
                   
                   
                 267.1667 
                 16.66667 
               
             
          
           
               
                 Fifth measurement 
               
             
          
           
               
                 A 
                 22 
                 12 
                 8 
                 384 
                 114.5 
                 22 
                 6 
                 6 
               
               
                 B 
                 22 
                 11 
                 8 
                 352 
                 172 
                 20 
                 4 
                 5 
               
               
                 C 
                 23 
                 12 
                 9 
                 486 
                 134 
                 20 
                 4 
                 4 
               
               
                 Mean 
                   
                   
                   
                 407.3333 
                 140.1667 
               
               
                   
               
             
          
           
               
                   
                 Low dosage (100 mg/kg/day) 
                 High dosage (200 mg/kg/day) 
               
             
          
           
               
                   
                   
                 Increased 
                   
                   
                   
                   
                 Increased 
               
               
                   
                 Volume 
                 tumor volume 
                 Weight 
                 L 
                 W 
                 Volume 
                 tumor volume 
               
               
                   
                 (mm 3 ) 
                 (mm 3 ) 
                 (g) 
                 (mm) 
                 (mm) 
                 (mm 3 ) 
                 (mm 3 ) 
               
               
                   
                   
               
             
          
           
               
                 First measurement 
               
             
          
           
               
                 A 
                 87.5 
                 87.5 
                 19 
                 6 
                 4 
                 48 
                 48 
               
               
                 B 
                 144 
                 144 
                 18.5 
                 8 
                 5 
                 100 
                 100 
               
               
                 C 
                 108 
                 108 
                 20 
                 7 
                 5 
                 87.5 
                 87.5 
               
               
                 Mean 
                 113.1667 
                 113.1667 
                   
                   
                   
                 78.5 
                 78.5 
               
             
          
           
               
                 Second measurement 
               
             
          
           
               
                 A 
                 75 
                 −12.5 
                 20 
                 6 
                 4 
                 48 
                 0 
               
               
                 B 
                 126 
                 −18 
                 23 
                 5 
                 3 
                 22.5 
                 −77.5 
               
               
                 C 
                 126 
                 18 
                 20 
                 7 
                 5 
                 87.5 
                 0 
               
               
                 Mean 
                 109 
                 −4.16667 
                   
                   
                   
                 52.66667 
                 −25.8333 
               
             
          
           
               
                 Third measurement 
               
             
          
           
               
                 A 
                 126 
                 51 
                 19 
                 5 
                 4 
                 40 
                 −8 
               
               
                 B 
                 56 
                 −70 
                 18.5 
                 0 
                 0 
                 0 
                 −22.5 
               
               
                 C 
                 87.5 
                 −38.5 
                 18.5 
                 0 
                 0 
                 0 
                 −87.5 
               
               
                 Mean 
                 89.83333 
                 −19.1667 
                   
                   
                   
                 13.33333 
                 −39.3333 
               
             
          
           
               
                 Fourth measurement 
               
             
          
           
               
                 A 
                 108 
                 −18 
                 20 
                 0 
                 0 
                 0 
                 −40 
               
               
                 B 
                 32 
                 −24 
                 21 
                 0 
                 0 
                 0 
                 0 
               
               
                 C 
                 75 
                 −12.5 
                 20 
                 0 
                 0 
                 0 
                 0 
               
               
                 Mean 
                 71.66667 
                 −18.1667 
                   
                   
                   
                 0 
                 −13.3333 
               
             
          
           
               
                 Fifth measurement 
               
             
          
           
               
                 A 
                 108 
                 0 
                 21 
                 0 
                 0 
                 0 
                 0 
               
               
                 B 
                 50 
                 18 
                 20 
                 0 
                 0 
                 0 
                 0 
               
               
                 C 
                 32 
                 −43 
                 20 
                 0 
                 0 
                 0 
                 0 
               
               
                 Mean 
                 63.33333 
                 −8.33333 
                   
                   
                   
                 0 
                 0 
               
               
                   
               
             
          
         
       
     
         [0033]    According to the result of  FIG. 2 , both low dosage and high dosage of duloxetine HCl have great inhibitory effect against the tumors. There was no obvious weight loss in each group of mice during the experiments. It thus indicates that duloxetine HCl, either high or low dosage, may let the test mice be in optimal health conditions and not die during the treatment. 
         [0034]    According to the result of  FIG. 3 , low dosage and high dosage of duloxetine HCl may effectively inhibit the increase of tumor volume, and meanwhile decrease the tumor volume. In particular, high dosage of duloxetine HCl has a better effect. 
         [0035]    The detailed description above serves to illustrate the practicable embodiment (s) of the present disclosure. However, such embodiment (s) cannot be utilized to limit the claimed scope of the present disclosure. Accordingly, the claimed scope of the present disclosure is intended to encompass all equivalent practice or changes without departing from the spirit of the invention.