Abstract:
A method for treating a cancer includes administering to a subject in need thereof a pharmaceutical composition containing a therapeutically effective amount of Azelnidipine or a pharmaceutical acceptable salt thereof. The cancer is selected from the group consisting of a pleural-related cancer, an abdominal-related cancer, an endocrine-related cancer, a gastrointestinal tract-related cancer, osteosarcoma, and skin cancer.

Description:
CROSS-REFERENCE TO RELATED APPLICATIONS 
       [0001]    This is a National Phase Application filed under 35 U.S.C. 371 as a national stage of PCT/CN2015/092714 filed Oct. 23, 2015, an application claiming the benefit under 35 USC 119(e) to the following U.S. Provisional Applications No. 62/068,298 filed Oct. 24, 2014, the content of each of which is hereby incorporated by reference in its entirety. 
     
    
     FIELD OF THE INVENTION 
       [0002]    The present invention related to a new indication of Azelnidipine pharmaceutical composition, especially related to inhibition effect of Azelnidipine pharmaceutical composition on a variety of cancer cells. 
       BACKGROUND OF THE INVENTION 
       [0003]    Azelnidipine is a drug for treating hypertension and can be used alone or in combination with other antihypertensive drugs. Azelnidipine is approved by FDA and accumulated a huge data of drug use and drug mechanism research. 
         [0004]    Due to the differences of the clinical use, there is no research present that the Azelnidipine has any potential to inhibit cancer cell. 
         [0005]    On the other side, cancer is the most popular disease cause of death in the world. The cancer patients are gradually increase yearly, therefore the treatment method of the cancer has become an important issue. The medical treatments of cancer can be classified as surgical treatment, radiation therapy, chemotherapy and target therapy. Generally, the cancer drug, whether chemotherapy drug or target therapy drug, is to inhibit cancer cells duplication and split to prevent the tumor growth and metastasis. Averagely, only about five of 10,000 new drugs can successfully enter the phase I of clinical trials. Furthermore, if the cancer patients happen the drug resistance, that would reduce the effectiveness of the drugs and result in the medical treatment failure. Therefore, the new drug development is very difficult. 
         [0006]    Therefore, it is a very urgent and important issue that how to develop anti-cancer drugs quickly and reduce the probability of clinical failure for treating various cancers. 
       SUMMARY OF THE INVENTION 
       [0007]    In order to solve the above problems, the present invention provide the development of new cancer clinical indications of Azelnidipine. 
         [0008]    Accordingly, the present invention provides a new indication of Azelnidipine. The experimental results showed that the Azelnidipine had no toxicity or had little toxicity to normal cells in the present invention. However, the selective effect of Azelnidipine between normal cells and cancer cells need to be identified. 
         [0009]    The present invention provides a pharmaceutical composition of Azelnidipine for treating cancer. The pharmaceutical composition is composed of effective dose of Azelnidipine and a pharmaceutical acceptable salt. 
         [0010]    In one embodiment of the present invention, the cancer is selected from pleural-related cancer, abdominal-related cancer, endocrine-related cancer, gastrointestinal tract-related cancer. 
         [0011]    In one embodiment of the present invention, the cancer is selected from osteosarcoma, skin cancer and blood cancer. 
         [0012]    In one embodiment of the present invention, the pleural-related cancer is lung cancer. 
         [0013]    In one embodiment of the present invention, the abdominal-related cancer is selected from bladder cancer, cervical cancer, and kidney cancer. 
         [0014]    In one embodiment of the present invention, the endocrine-related cancer is selected from prostate cancer, breast cancer, and ovarian cancer. 
         [0015]    In one embodiment of the present invention, the gastrointestinal tract-related cancer is selected from gastric cancer, hepatic cancer, colorectal cancer, pancreatic cancer, and tongue cancer. 
         [0016]    In one embodiment of the present invention, the effective dose of Azelnidipine is from 2.0 mg/kg/day to 500 mg/kg/day. 
     
    
     
       BRIEF DESCRIPTION OF THE DRAWINGS 
         [0017]      FIG. 1  shows the results of the inhibitory effect of the different cancer cells by Azelnidipine. 
           [0018]      FIG. 2  shows the results of the inhibitory effect of tumor volume by Azelnidipine. 
           [0019]      FIG. 3  shows the inhibitory effect of tumor growth via administered high-dose and low-dose of Azelnidipine. 
       
    
    
     DETAILED DESCRIPTION OF THE INVENTION 
       [0020]    Cell Culture 
         [0021]    Subculture the different types of cancer cell lines. The cancer cells includes lung cancer, gastric cancer, hepatic cancer, colon cancer, skin cancer, cervical cancer, prostate cancer, bladder cancer, breast cancer, leukemia, pancreatic cancer, ovarian cancer, tongue cancer, osteosarcoma, and renal cancer. The normal cells used in the control group included kidney cell (HEK293) and human bronchial epithelial cell line BEAS-2B (as shown in Table 1). 
         [0022]    Cancer cell lines were cultured in different culture medium according to different characteristics (as shown in Table 1). The cell numbers were counted and reseed as 2×10 6  in cell culture plate/flask. Then, the culture medium were added to a volume of 10 ml, and the cells were cultured for 2-3 days. Then, the cells were suspended for loading into 96-well plates. The number of cells was 3000 and the volume of the culture medium was 100 μl each well. 
         [0000]    
       
         
               
             
               
               
               
               
             
               
               
               
               
             
           
               
                 TABLE 1 
               
             
             
               
                   
               
               
                 Cancer cell lines and the culture medium 
               
             
          
           
               
                   
                   
                   
                 Culture 
               
               
                 No 
                 Cancer type 
                 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 
                 hepatic 
                 HepG2 (hepatocellular carcinoma) 
                 DMEM 
               
               
                   
                 cancer 
                 Hep3B (hepatocellular carcinoma) 
                 DMEM 
               
               
                 4 
                 colon 
                 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) 
                 MEM 
               
               
                   
                   
                 BCRC60554 
               
               
                 7 
                 prostate 
                 PC3 (p53−)(Prostate adenocarcinoma) 
                 DMEM 
               
               
                   
                 cancer 
                 LNCaP clone FGC (LNCap.FGC) 
                 RPMI-1640 
               
               
                 8 
                 bladder 
                 8301 (urinary bladder carcinoma) 
                 RPMI-1640 
               
               
                   
                 cancer 
                 T24 
                 RPMI-1640 
               
               
                 9 
                 breast 
                 MCF7 (Mammary Gland, 
                 DMEM 
               
               
                   
                 cancer 
                 Adenocarcinoma) 
                 DMEM 
               
               
                   
                   
                 MDA-MB-231 (Mammary Gland, 
               
               
                   
                   
                 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 
                 tongue 
                 SAS (Tongue squamous cell carcinoma) 
                 DMEM 
               
               
                   
                 cancer 
               
               
                 13 
                 osteo- 
                 U-2OS 
                 DMEM 
               
               
                   
                 sarcoma 
               
               
                 14 
                 renal 
                 786-O (Renal adenocarcinoma) 
                 RPMI-1640 
               
               
                   
                 cancer 
                 BCRC 60243 
               
               
                 15 
                 normal cell 
                 HEK293 (Kidney) 
                 DMEM 
               
               
                   
                 kidney 
                 BEAS-2B (Lung Epithelial) 
                 RPMI-1640 
               
               
                   
                 pulmonary 
               
               
                   
                 epithelial 
               
               
                   
                 cell line 
               
               
                   
               
             
          
         
       
     
         [0023]    Cell Viability Analysis 
         [0024]    Removing the original culture medium from 96-well plate. Then add 100 μl of commercially drug at a concentration of 10 μM per well. After 72 hours, add the diluted WST-1 reagent to the well with 100 μl/well, and the diluted WST-1 reagent was acquired from the dilution of 9:1 medium and WST-1 stock reagent. Finally, the total volume of each well was 200 μl/well. Culture the 96-well plate at 37° C. for 30 to 90 minutes. Detecting and calculate the survival rate of each cancer cells with an ELISA reader at OD450 nm. The lower viability of cancer cells represents better inhibition effect via the Azelnidipine drug. Otherwise, the higher viability of cancer cells represents worse inhibition effect via the Azelnidipine drug. 
         [0025]    The Effect of Azelnidipine on Different Cancer Cell Lines 
         [0026]    The Inhibition Effect of Azelnidipine on Pleural-Related Cancer Cells 
         [0027]    This inhibition test of Azelnidipine on pleural-related cancer cells were using two lung cancer cell lines A549 and H1650. The inhibitory tests of Azelnidipine were performed 4 times for each cell lines and then the average value of the inhibitory tests was calculated. The results were shown in Table 2. 
         [0000]    
       
         
               
             
               
               
               
               
               
               
               
             
               
               
               
               
               
               
               
             
           
               
                 TABLE 2 
               
               
                   
               
               
                 The inhibition effect of Azelnidipine on pleural-related cancer cells 
               
               
                   
               
             
             
               
                   
               
             
          
           
               
                   
                   
                 0524-10 
                 0526-10 
                 0529-10 
                 0531-10 
                   
               
               
                   
                   
                 min 
                 min 
                 min 
                 min 
                 Average 
               
               
                   
                   
               
               
                   
                 A549 
                 81.4 
                 118.6 
                 69.7 
                 80.2 
                 87.5 
               
               
                   
                   
               
             
          
           
               
                   
                   
                 1-10 
                 2-20 
                 3-20 
                 4-20 
                   
               
               
                   
                   
                 min 
                 min 
                 min 
                 min 
                 Average 
               
               
                   
                   
               
               
                   
                 H1650 
                 28.8 
                 26.5 
                 40.5 
                 62.7 
                 39.7 
               
               
                   
                   
               
             
          
         
       
     
         [0028]    The Inhibition Effect of Azelnidipine on Abdominal-Related Cancer Cell Lines 
         [0029]    This inhibition test of Azelnidipine on abdominal-related cancer cells were using bladder cancer cell line TSGH and T24 (Table 3), cervical cancer cell lines HeLa and C-33A (Table 4), renal cancer cell line 786-O (Table 5). The inhibitory tests of Azelnidipine were performed 4 times for each cell lines and then the average value of the inhibitory tests was calculated. The results were shown in Table 3, Table 4, and Table 5. 
         [0000]    
       
         
               
             
               
               
               
               
               
               
             
               
               
               
               
               
               
               
             
           
               
                 TABLE 3 
               
               
                   
               
               
                 The inhibition effect of Azelnidipine on bladder cancer cell lines 
               
               
                   
               
             
             
               
                   
               
             
          
           
               
                   
                 0510-10 
                 0512-10 
                 0515-10 
                 0517-10 
                   
               
               
                   
                 min 
                 min 
                 min 
                 min 
                 average 
               
               
                   
               
               
                 TSGH 
                 151.0 
                 117.3 
                 72.0 
                 120.1 
                 115.1 
               
               
                   
               
             
          
           
               
                   
                   
                 1-30 
                 2-20 
                 3-20 
                 4-20 
                   
               
               
                   
                   
                 min 
                 min 
                 min 
                 min 
                 average viability 
               
               
                   
                   
               
               
                   
                 T24 
                 109.7 
                 75.3 
                 30.4 
                 102.5 
                 79.5 
               
               
                   
                   
               
             
          
         
       
     
         [0000]    
       
         
               
             
               
               
               
               
               
               
             
               
               
               
               
               
               
             
           
               
                 TABLE 4 
               
             
             
               
                   
               
               
                 The inhibition effect of Azelnidipine on cervical cancer cell lines 
               
             
          
           
               
                   
                 0524-10 
                 0526-10 
                 0529-10 
                 0531-10 
                   
               
               
                   
                 min 
                 min 
                 min 
                 min 
                 Average 
               
               
                   
                   
               
             
          
           
               
                 HeLa 
                 70.4 
                 93.7 
                 81.4 
                 102.7 
                 87.0 
               
               
                 C-33A 
                 141.8 
                 110.6 
                 123.4 
                 117.4 
                 123.3 
               
               
                   
               
             
          
         
       
     
         [0000]    
       
         
               
             
               
               
               
               
               
               
             
               
               
               
               
               
               
             
           
               
                 TABLE 5 
               
             
             
               
                   
               
               
                 The inhibition effect of Azelnidipine on renal cancer cell line 
               
             
          
           
               
                   
                 0524-10 
                 0526-10 
                 0529-10 
                 0531-10 
                   
               
               
                   
                 min 
                 min 
                 min 
                 min 
                 average 
               
               
                   
                   
               
             
          
           
               
                 786-O 
                 88.6 
                 79.7 
                 66.5 
                 112.5 
                 86.8 
               
               
                   
               
             
          
         
       
     
         [0030]    The Inhibition Effect of Azelnidipine on Endocrine-Related Cancer Cell Lines 
         [0031]    This inhibition test of Azelnidipine on endocrine-related cancer cells were using prostate cancer cell line PC-3 (Table 6), breast cancer cell lines MCF7 and MDA-MB-231 (Table 7), and ovarian cancer cell lines NIH-OVCAR-3 and TOV-21G (Table 8). The inhibitory tests of Azelnidipine were performed 4 times for each cell lines and then the average value of the inhibitory tests was calculated. The results were shown in Table 6, Table 7, and Table 8. 
         [0000]    
       
         
               
             
               
               
               
               
               
               
             
               
               
               
               
               
               
             
           
               
                 TABLE 6 
               
             
             
               
                   
               
               
                 The inhibition effect of Azelnidipine on prostate cancer cell line 
               
             
          
           
               
                   
                 PC-3-0524- 
                 PC-3-0526- 
                 PC-3-0529- 
                 PC-3-0531- 
                 Aver- 
               
               
                   
                 10 min 
                 10 min 
                 10 min 
                 10 min 
                 age 
               
               
                   
                   
               
             
          
           
               
                 PC-3 
                 122.1 
                 149.4 
                 110.2 
                 102.0 
                 120.9 
               
               
                   
               
             
          
         
       
     
         [0000]    
       
         
               
             
               
               
               
               
               
               
             
               
               
               
               
               
               
             
           
               
                 TABLE 7 
               
               
                   
               
               
                 The inhibition effect of Azelnidipine on breast cancer cell lines 
               
               
                   
               
             
             
               
                   
               
             
          
           
               
                   
                 0612-10 
                 0614-10 
                 0616-10 
                 0619-10 
                   
               
               
                   
                 min 
                 min 
                 min 
                 min 
                 average 
               
               
                   
               
               
                 MCF7 
                 48.6 
                 46.6 
                 68.2 
                 79.9 
                 60.8 
               
               
                   
               
             
          
           
               
                   
                   
                 0612-10 
                 0614-10 
                 0616-10 
                   
               
               
                   
                   
                 min 
                 min 
                 min 
                 average 
               
               
                   
                   
               
               
                   
                 MDA-MB-231 
                 70.3 
                 89.7 
                 100.8 
                 86.9 
               
               
                   
                   
               
             
          
         
       
     
         [0000]    
       
         
               
             
               
               
               
               
               
               
             
               
               
               
               
               
               
             
           
               
                 TABLE 8 
               
               
                   
               
               
                 The inhibition effect of Azelnidipine on ovarian cancer cell lines 
               
               
                   
               
             
             
               
                   
               
             
          
           
               
                   
                 7-3-30 
                 7-4-30 
                 7-7-30 
                 7-4-30 
                   
               
               
                   
                 min 
                 min 
                 min 
                 min 
                 average 
               
               
                   
               
               
                 NIH-OVCAR-3 
                 106.3 
                 99.1 
                 100.9 
                 103.5 
                 102.4 
               
               
                   
               
             
          
           
               
                   
                 7-3-30 
                 7-4-30 
                 7-7-30 
                 4-30 
                   
               
               
                   
                 min 
                 min 
                 min 
                 min 
                 average 
               
               
                   
               
               
                 TOV-21G 
                 15.6 
                 13.3 
                 19.6 
                 60.1 
                 27.2 
               
               
                   
               
             
          
         
       
     
         [0032]    The Inhibition Effect of Azelnidipine on Gastrointestinal Tract-Related Cancer Cell Lines 
         [0033]    This inhibition test of Azelnidipine on gastrointestinal tract-related cancer cells were using gastric cancer cell lines AGS and MKN-45 (Table 9), hepatic cancer cell lines HepG2 and Hep3B (Table 10), colorectal cancer cell lines HCT116-wt and LoVo (Table 11), pancreatic cancer cell lines AsPC-1 and BxPC-3 (Table 12), tongue cancer cell line SAS (Table 13). The inhibitory tests of Azelnidipine were performed 4 times for each cell lines and then the average value of the inhibitory tests was calculated. The results were shown in Table 9, Table 10, Table 11, Table 12 and Table 13. 
         [0000]    
       
         
               
             
               
               
               
               
               
               
               
             
               
               
               
               
               
               
             
           
               
                 TABLE 9 
               
               
                   
               
               
                 The inhibition effect of Azelnidipine on gastric cancer cell lines. 
               
               
                   
               
             
             
               
                   
               
             
          
           
               
                   
                   
                 0510-10 
                 0512-10 
                 0515-10 
                 0517-10 
                   
               
               
                   
                   
                 min 
                 min 
                 min 
                 min 
                 average 
               
               
                   
                   
               
               
                   
                 AGS 
                 120.2 
                 124.3 
                 42.1 
                 74.3 
                 90.2 
               
               
                   
                   
               
             
          
           
               
                   
                 0510-10 
                 0512-10 
                 0515-10 
                 0517-10 
                   
               
               
                   
                 min 
                 min 
                 min 
                 min 
                 Average 
               
               
                   
               
               
                 MKN-45 
                 173.5 
                 183.7 
                 68.5 
                 130.3 
                 139.0 
               
               
                   
               
             
          
         
       
     
         [0000]    
       
         
               
             
               
               
               
               
               
               
             
           
               
                 TABLE 10 
               
               
                   
               
               
                 The inhibition effect of Azelnidipine on hepatic cancer cell lines 
               
               
                   
               
             
             
               
                   
               
             
          
           
               
                   
                 0524-20 
                 0526-20 
                 0529-20 
                 0531-20 
                   
               
               
                   
                 min 
                 min 
                 min 
                 min 
                 Average 
               
               
                   
               
               
                 HepG2 
                 110.3 
                 87.2 
                 112.1 
                 93.7 
                 100.8 
               
               
                   
               
               
                   
                 0612-20 
                 0614-20 
                 0616-20 
                 0619-20 
               
               
                   
                 min 
                 min 
                 min 
                 min 
                 Average 
               
               
                   
               
               
                 Hep3B 
                 114.2 
                 91.5 
                 97.9 
                 90.7 
                 98.6 
               
               
                   
               
             
          
         
       
     
         [0000]    
       
         
               
             
               
               
               
               
               
               
             
               
               
               
               
               
               
               
             
           
               
                 TABLE 9 
               
               
                   
               
               
                 The inhibition effect of Azelnidipine on colorectal cancer cell lines 
               
               
                   
               
             
             
               
                   
               
             
          
           
               
                   
                 0602-30 
                 0605-10 
                 0607-10 
                 0609-10 
                   
               
               
                   
                 min 
                 min 
                 min 
                 min 
                 Average 
               
               
                   
               
               
                 HCT116-wt 
                 70.7 
                 146.2 
                 158.9 
                 69.3 
                 111.3 
               
               
                   
               
             
          
           
               
                   
                   
                 0616-10 
                 0619-10 
                 0621-10 
                 0623-10 
                   
               
               
                   
                   
                 min 
                 min 
                 min 
                 min 
                 Average 
               
               
                   
                   
               
               
                   
                 LoVo 
                 49.6 
                 67.2 
                 62.3 
                 60.3 
                 59.9 
               
               
                   
                   
               
             
          
         
       
     
         [0000]    
       
         
               
             
               
               
               
               
               
               
             
           
               
                 TABLE 12 
               
               
                   
               
               
                 The inhibition effect of Azelnidipine on pancreatic cancer cell lines 
               
               
                   
               
             
             
               
                   
               
             
          
           
               
                   
                 1-7-3-30 
                 1-7-4-30 
                 1-7-7-30 
                 1-4-30 
                   
               
               
                   
                 min 
                 min 
                 min 
                 min 
                 Average 
               
               
                   
               
               
                 AsPC-1 
                 103.9 
                 46.1 
                 13.7 
                 58.6 
                 55.6 
               
               
                   
               
               
                   
                 3-7-3-30 
                 3-7-4-30 
                 3-7-7-30 
                 3-4-30 
               
               
                   
                 min 
                 min 
                 min 
                 min 
                 Average 
               
               
                   
               
               
                 BxPC-3 
                 79.9 
                 51.0 
                 50.0 
                 77.5 
                 64.6 
               
               
                   
               
             
          
         
       
     
         [0000]    
       
         
               
             
               
               
               
               
               
               
             
               
               
               
               
               
               
               
             
           
               
                 TABLE 13 
               
             
             
               
                   
               
               
                 The inhibition effect of Azelnidipine on tongue cancer cell line 
               
             
          
           
               
                   
                 6-26-10 
                 6-28-10 
                 6-30-10 
                 7-3-10 
                   
               
               
                   
                 min 
                 min 
                 min 
                 min 
                 Average 
               
               
                   
                   
               
             
          
           
               
                   
                 SAS 
                 69.9 
                 66.0 
                 66.7 
                 104.7 
                 76.8 
               
               
                   
                   
               
             
          
         
       
     
         [0034]    The Inhibition Effect of Azelnidipine on Other Cancer Cell Lines 
         [0035]    This inhibition test of Azelnidipine on other cancer cells were using osteosarcoma cell line U2OS (Table 14), skin cancer cell lines A375 and BCC (Table 15). The inhibitory tests of Azelnidipine were performed 3 to 4 times for each cell lines and then the average value of the inhibitory tests was calculated. The results were shown in Table 14 and Table 15. 
         [0000]    
       
         
               
             
               
               
               
               
               
               
             
               
               
               
               
               
               
             
           
               
                 TABLE 14 
               
             
             
               
                   
               
               
                 The inhibition effect of Azelnidipine 
               
               
                 on osteosarcoma cancer cell line 
               
             
          
           
               
                   
                 6-26-10 
                 6-28-10 
                 6-30-10 
                 7-3-10 
                   
               
               
                   
                 min 
                 min 
                 min 
                 min 
                 Average 
               
               
                   
                   
               
             
          
           
               
                 U2OS 
                 53.3 
                 78.3 
                 84.8 
                 61.2 
                 69.4 
               
               
                   
               
             
          
         
       
     
         [0000]    
       
         
               
             
               
               
               
               
               
               
               
             
               
               
               
               
               
               
             
           
               
                 TABLE 15 
               
               
                   
               
               
                 The inhibition effect of Azelnidipine on skin cancer cell lines 
               
               
                   
               
             
             
               
                   
               
             
          
           
               
                   
                   
                 0602-30 
                 0605-10 
                 0607-10 
                 0609-10 
                   
               
               
                   
                   
                 min 
                 min 
                 min 
                 min 
                 Average 
               
               
                   
                   
               
               
                   
                 A375 
                 123.6 
                 100.2 
                 76.0 
                 104.9 
                 101.2 
               
               
                   
                   
               
             
          
           
               
                   
                   
                 0602-30 
                 0605-10 
                 0607-10 
                   
               
               
                   
                   
                 min 
                 min 
                 min 
                 Average 
               
               
                   
                   
               
               
                   
                 BCC 
                 77.7 
                 127.2 
                 95.7 
                 100.2 
               
               
                   
                   
               
             
          
         
       
     
         [0036]    The Experiment Design on Control Group 
         [0037]    The Inhibition Effect of Azelnidipine on Normal Cells 
         [0038]    This inhibition test of Azelnidipine on normal cells were using normal kidney cell line HEK293 (Table 16) and normal pulmonary epithelial cell lines BEAS-2B (Table 17). The inhibitory tests of Azelnidipine were performed 4 times for each cell lines and then the average value of the inhibitory tests was calculated. The results were shown in Table 16 and Table 17. 
         [0000]    
       
         
               
             
               
               
             
               
               
               
             
           
               
                 TABLE 16 
               
             
             
               
                   
               
               
                 The inhibition effect of Azelnidipine on normal kidney cell line 
               
             
          
           
               
                   
                 Average 
               
               
                   
                   
               
             
          
           
               
                   
                 HEK293 
                 94.2 
               
               
                   
                   
               
             
          
         
       
     
         [0000]    
       
         
               
             
               
               
               
               
               
               
             
               
               
               
               
               
               
             
           
               
                 TABLE 17 
               
             
             
               
                   
               
               
                 The inhibition effect of Azelnidipine on 
               
               
                 normal pulmonary epithelial cell line 
               
             
          
           
               
                   
                 0510-10 
                 0512-10 
                 0515-10 
                 0517-10 
                   
               
               
                   
                 min 
                 min 
                 min 
                 min 
                 average 
               
               
                   
                   
               
             
          
           
               
                 BEAS-2B 
                 61.3 
                 61.8 
                 101.6 
                 85.4 
                 77.5 
               
               
                   
               
             
          
         
       
     
         [0039]    This inhibition test results of Azelnidipine on all kinds of cells were shown in Table 18. It is clear that Azelnidipine has a significant inhibitory effect on several cancers cell lines. As a result in the experiments of the present invention. Paroxetine has a significant inhibitory effect on various cancer cells ( FIG. 1 ). 
         [0000]    
       
         
               
             
               
               
             
               
               
               
             
           
               
                 TABLE 18 
               
             
             
               
                   
               
               
                 Summary of the Effect on different 
               
               
                 cancer cell lines by Azelnidipine 
               
             
          
           
               
                   
                 Inhibitory effect 
               
               
                   
                   
               
             
          
           
               
                   
                 cancer cells 
                   
               
               
                   
                 lung cancer 
                 63.60 
               
               
                   
                 bladder cancer 
                 97.30 
               
               
                   
                 cervical cancer 
                 87.00 
               
               
                   
                 Kidney cancer 
                 86.80 
               
               
                   
                 prostate cancer 
                 60.45 
               
               
                   
                 breast cancer 
                 73.85 
               
               
                   
                 ovarian cancer 
                 64.80 
               
               
                   
                 gastric cancer 
                 114.60 
               
               
                   
                 hepatic cancer 
                 99.70 
               
               
                   
                 colorectal cancer 
                 85.60 
               
               
                   
                 pancreatic cancer 
                 60.10 
               
               
                   
                 tongue cancer 
                 76.80 
               
               
                   
                 osteosarcoma 
                 69.40 
               
               
                   
                 skin cancer 
                 100.70 
               
               
                   
                 Normal cell 
               
               
                   
                 Kidney cell 
                 94.20 
               
               
                   
                 pulmonary epithelial cell line 
                 77.50 
               
               
                   
                   
               
             
          
         
       
     
         [0040]    Animal Model Test of Ovarian Cancer with Dose 100 mg/kg/Day and 200 mg/kg/day 
         [0041]    In this invention, the female mice were (BALB/cAnN.Cg-Foxn1 nu /CrlNarl) purchased from National Laboratory Animal Center (Taiwan)). The weight of the mice were 21±1 g. These mice were subcutaneously injected with gastric cancer cells (AGS) and then put these mice into different cages at random. The drug test experiment was divided into three groups, include “normal control group”, “low dose group (100 mg/kg/day)”, and “high dose group (200 mg/kg/day)”. These mice were then injected test drug intraperitoneally once daily until the tumor size reached 100 mm 3 . The tumor sizes and body weight were measured twice a week. The tumor sizes were measured and calculated by formula: (L×W 2 )/2. L represents the tumor longest length. W represents the tumor shortest diameter. The experiment results were shown in Table 19 
         [0000]    
       
         
               
             
               
               
               
               
             
               
               
               
               
               
               
               
               
               
               
               
               
               
               
               
               
             
               
             
               
               
               
               
               
               
               
               
               
               
               
               
               
               
               
               
             
               
             
               
               
               
               
               
               
               
               
               
               
               
               
               
               
               
               
             
               
             
               
               
               
               
               
               
               
               
               
               
               
               
               
               
               
               
             
               
             
               
               
               
               
               
               
               
               
               
               
               
               
               
               
               
               
             
               
             
               
               
               
               
               
               
               
               
               
               
               
               
               
               
               
               
             
           
               
                 TABLE 19 
               
             
             
               
                   
               
               
                 The inhibitory effect of tumor volume via administered Azelnidipine 
               
             
          
           
               
                   
                 control group 
                 low dose (100 mg/kg/day) 
                 high dose (200 mg/kg/day) 
               
             
          
           
               
                   
                   
                   
                   
                   
                 tumor 
                   
                   
                   
                   
                 tumor 
                   
                   
                   
                   
                 tumor 
               
               
                   
                   
                 longest 
                   
                 vol- 
                 volume 
                   
                 longest 
                   
                   
                 volume 
                   
                 longest 
                   
                   
                 volume 
               
               
                   
                 weight 
                 length 
                 width 
                 ume 
                 growth 
                 weight 
                 length 
                 width 
                 volume 
                 growth 
                 weight 
                 length 
                 width 
                 volume 
                 growth 
               
               
                   
                 (g) 
                 mm 
                 mm 
                 mm 3   
                 mm 3   
                 (g) 
                 mm 
                 mm 
                 mm 3   
                 mm 3   
                 (g) 
                 mm 
                 mm 
                 mm 3   
                 mm 3   
               
               
                   
                   
               
             
          
           
               
                 First measurement 
               
             
          
           
               
                 A 
                 18.5 
                 7 
                 7 
                 171.5 
                 171.5 
                 21 
                 8 
                 8 
                 256 
                 256 
                 20 
                 4 
                 3 
                 18 
                 18 
               
               
                 B 
                 22 
                 8 
                 6 
                 144 
                 144 
                 21 
                 6 
                 7 
                 147 
                 147 
                 19.5 
                 6 
                 3 
                 27 
                 27 
               
               
                 C 
                 20.5 
                 9 
                 8 
                 288 
                 288 
                 21 
                 7 
                 6 
                 126 
                 126 
                 20 
                 4 
                 4 
                 32 
                 32 
               
               
                 av- 
                 20.4 
                 7.6 
                 7 
                 189.3 
                 189.3 
                   
                   
                   
                 176.3333 
                 176.3333 
                   
                   
                   
                 25.66667 
                 25.66667 
               
               
                 er- 
               
               
                 age 
               
             
          
           
               
                 Second measurement 
               
             
          
           
               
                 A 
                 22 
                 7 
                 6 
                 126 
                 −45.5 
                 20 
                 7 
                 6 
                 126 
                 −130 
                 19 
                 7 
                 5 
                 87.5 
                 69.5 
               
               
                 B 
                 20 
                 8 
                 7 
                 196 
                 52 
                 20 
                 6 
                 6 
                 108 
                 −39 
                 20 
                 6 
                 5 
                 75 
                 48 
               
               
                 C 
                 20 
                 9 
                 7 
                 220.5 
                 −67.5 
                 19 
                 5 
                 5 
                 62.5 
                 −63.5 
                 19 
                 7 
                 5 
                 87.5 
                 55.5 
               
               
                 av- 
                 20.6 
                 8.4 
                 6.8 
                 198.5 
                 9.2 
                   
                   
                   
                 98.83333 
                 −77.5 
                   
                   
                   
                 83.33333 
                 57.66667 
               
               
                 er- 
               
               
                 age 
               
             
          
           
               
                 Third measurement 
               
             
          
           
               
                 A 
                 23 
                 9 
                 6 
                 162 
                 36 
                 19.5 
                 7 
                 6 
                 126 
                 0 
                 20.5 
                 7 
                 5 
                 87.5 
                 0 
               
               
                 B 
                 20 
                 10 
                 8 
                 320 
                 124 
                 19 
                 6 
                 6 
                 108 
                 0 
                 19 
                 5 
                 5 
                 62.5 
                 −12.5 
               
               
                 C 
                 21 
                 11 
                 7 
                 269.5 
                 49 
                 18.5 
                 5 
                 5 
                 62.5 
                 0 
                 20 
                 0 
                 0 
                 0 
                 −87.5 
               
               
                 av- 
                 21.2 
                 10 
                 6.8 
                 235.3 
                 36.8 
                   
                   
                   
                 98.83333 
                 0 
                   
                   
                   
                 50 
                 −33.3333 
               
               
                 er- 
               
               
                 age 
               
             
          
           
               
                 Fourth measurement 
               
             
          
           
               
                 A 
                 23 
                 11 
                 7 
                 269.5 
                 107.5 
                 20 
                 4 
                 3 
                 18 
                 −108 
                 20 
                 0 
                 0 
                 0 
                 −87.5 
               
               
                 B 
                 22 
                 10 
                 6 
                 180 
                 −140 
                 21 
                 5 
                 4 
                 40 
                 −68 
                 20 
                 0 
                 0 
                 0 
                 −62.5 
               
               
                 C 
                 23 
                 11 
                 8 
                 352 
                 82.5 
                 20 
                 0 
                 0 
                 0 
                 −88 
                 21 
                 0 
                 0 
                 0 
                 0 
               
               
                 av- 
                 22.4 
                   
                   
                 233.5 
                 −1.8 
                   
                   
                   
                 19.33333 
                 −88 
                   
                   
                   
                 0 
                 −50 
               
               
                 er- 
               
               
                 age 
               
             
          
           
               
                 Fifth measurement 
               
             
          
           
               
                 A 
                 22 
                 12 
                 8 
                 384 
                 114.5 
                 20 
                 4 
                 3 
                 18 
                 0 
                 20 
                 0 
                 0 
                 0 
                 0 
               
               
                 B 
                 22 
                 11 
                 8 
                 352 
                 172 
                 20 
                 6 
                 4 
                 48 
                 8 
                 20 
                 0 
                 0 
                 0 
                 0 
               
               
                 C 
                 23 
                 12 
                 9 
                 486 
                 134 
                 21 
                 0 
                 0 
                 0 
                 0 
                 21 
                 0 
                 0 
                 0 
                 0 
               
               
                 av- 
                 22.4 
                   
                   
                 295.7 
                 62.2 
                   
                   
                   
                 22 
                 2.666667 
                   
                   
                   
                 0 
                 0 
               
               
                 er- 
               
               
                 age 
               
               
                   
               
             
          
         
       
     
         [0042]    According to the results in  FIG. 2 , low dose and high dose of Azelnidipine had significant inhibition effect on tumor cells. In the meantime, the weight of mice did not show a significant decrease during the experiment. These results indicated that both high and low doses of Azelnidipine could keep the tested mice in healthy status during the treatment without death. 
         [0043]    According to the results in  FIG. 3 , low dose and high dose of Azelnidipine had effectively slow down the tumor volume growth, and can also reduce the tumor volume. Especially, high dose of Azelnidipine had better effect to inhibit tumor growth. 
         [0044]    Although the present invention has been described in terms of specific exemplary embodiments and examples, it will be appreciated that the embodiments disclosed herein are for illustrative purposes only and various modifications and alterations might be made by those skilled in the art without departing from the spirit and scope of the invention as set forth in the following claims.