Experimental Study On Anti-tumor Effect Of Modified Amantadine

Malignant tumor is one of the main diseases which harms human's health. The traditional chemotherapeutic methods are usually going with great side effect, which hindered the anti-cancer chemicals'progress and development. Looking for the high performance, high specificity anti-cancer chemicals for specific cancers will be a new orientation for the anti-cancer chemicals'study at present. Amantadine is a traditional anti-influenza virus agent. According to some reports, poly-adamantane which is the analog of amantadine has anticancer effect. The forming of primary hepatic carcinoma (PHC) has really tight relationship with the history of viral hepatitis, about 80%~90% hepatoma patients had the history of type B or C hepatitis.We study the effect of modified amantadine (MA)on anti-cancer effect both in vivo and vitro to disclose it's anti-cancer mechanism.1. The toxicities of MA in vitro:MTT method was used to examine the cytotoxicity of MA in human normal FL cell. The result showed that no toxic activeties in low concentration except the concentration of 750μg /mL and 500μg/mL.It means that the MA has relatively low toxicity for human normal cell.2. The anti-cancer activity of MA in vitro:MTT method was used to examine the anti-cancer activity of MA on the human cancer cell lines of SMMC-7721 and SGC-7901.The result showed that MA could inhibit the growth and proliferation of human cancer cells, especially for the human cancer cell lines of SMMC-7721 with a great inhibition activity.3. The toxicity of MA in vivo:The acute toxicity test was used to detect the toxicity of MA by mouth in mice. The result showed that the 50% lethal dose (LD50) of MA is 810.03 mg/kg and the range of 95% confident limit is (715.15～917.49)mg/kg. It means that MA is a low toxic material.4. The anti-tumor activity in vivo:the tumor bearing mice with hepatoma-22 was established for anti-tumor experiment. The groups of MA were divided into 3 groups, they were low dose group, middle dose group and high dose group. After 10 days treatment with MA by ip (intraperitoneal injection), low, middle, and high dose groups inhibition rate (IR) in H22 were 31.01%, 36.05%, 51.94% respectively. The IR in high dose group is not lower than 5-Fu group. It means that MA has relatively high anti-cancer activity, and the high dose group has the similar IR with 5-Fu group.5. The activity of MA on immune system in mice:We detected the WBC value, the spleen and thymus index, the blastisation of lymphocyte test and the flow cytometry to test the activity of MA on immune system in mice.The result of WBC value showed that, MA couldn't decrease the value of WBC, MA has no toxicity effect on normal cells.The result of The organ index of all concentration groups of MA were not significantly compared with control group. The thymus index of 5-Fu group was significant lower than control group. It means that MA has no significant toxic effect on mice spleen and thymus in a limited range of concentration.The Result of blastisation of lymphocyte test was observed by the activity of spleen cell to mitosis ConA test. The result showed that the middle concentration group of MA had a significantly difference compared with control group. The low and high concentration groups of MA had no significant different compared with control group. It means that MA could increase the growth of lymphocyte, has no toxicity to the immune system.The FCM on tumor bearing mice spleen result showed that the apoptosis rate of low concentration group is significantly different compared with the control group. The G0/G1 stage cell of the low concentration group is significantly higher compared with the control group. It means that the middle dose group of MA couldn't block the mice spleen cells'cell cycle. MA couldn't conduct the apoptosis of spleen cell in mice.All the results showed that MA has efficient anti-cancer effect both in vivo and in vitro. MA belongs to the low toxic material. The mechanism of anti-cancer activity of MA perhaps is that it helps to strengthen the activity of immunity, protect immune organs and increase the immune activity to conduct the anti-cancer function. But we need to do more experiments to disclose the deeper mechanism of MA on anti-cancer function.