Preparation Technology And Process; Pharmacodynamics And Quality Standard Of Cinobutacini For Injection

Tumor is a kind of familiar disease,and with the speed up of aging and pollution,it becomes the most serious disease that endangers the people's health. The death rate of tumor sufferers is a little more than that of cardiopathy. This makes it the first killer of human health.The main purpose of this article mainly works over the new Medicine on its prescription, manufacture technics,quality standard and pharmacology according to the request of the National New Drug Examination and Approval Method. The research of pharmaceutical included pharmaceutical technology,quality standard and preliminary stability three parts and the achieving data establish academic.Studied the extract process of preparation, developed an optimal process of extraction with traditional medicated process and modern technique,the method is water extraction and ethanol precipitation,water washing-out supernumerary substance in polyamide column,collect ethanol elution. Chosen orthogonal table L9(34)in the experiment of decoction technology to research the productivity of cinobutacini from technological parameters example the time of the herb marinating,the volume of water filled,extraction times,the time of extraction etc.Orthogonal test applied to water-extracting process of cinobutacini,alkaloid extracting rate used for reference index, spectrophotometer was used for the content assaying. The best technique was two times water and forty minute for decoction in first extracting process, ten times water and forty minute in second extracting process. It is obey the rule of Chinese Medical, extract the effective component and simplify the process at the same time. Above all, the best abstract technology of vapour distillation was determined. Cinobutacini lyophilized powder was made by freeze-drying.The concentration of cinobutacini solution and the varieties and quantities of filling material were selected as well as the stability of cinobutacini solution and powder for injection were examined primarily.The quality of Cinobutacini Lyophilized Powder is more stable than that of solution for injection.Cinobutacini for Injection quality standard was established, cinobutacini for Injection is steady. The result shows that this optimized process is stable and feasible.On the other hand, according to the guide rules of Chinese medical injection in new medicine evaluation, to make sure this injection is safety, effective and stability, assurance the quality of the injection and found the base of injection's fingerprint.The method is simple,accurate reproducible and can be applied for quality control. Research of the quantity standard has been completed, standard of drug and preparation has been set up,the result show this method is simple and feasible and can guarantee the stability of this preparation.Preliminary stability research:The cinobutacini for Injection was determined by the above methods,results indicated that cinobutacini for Injection is steady in six months. To study the inhibitory effect of cinobutacini on transplantation tumors and the effect on immune function in mice .The models of mice S180 sarcoma and mice transplanted tumor H22 and Lewis were used to test the anti-tumor effect in vivo.To establish the animal model of lung carcinoma. The subculturing Lewis lung cancer cells (LLC) were inoculated subcutaneously to the axilla of C57BL/6 mice to make models .The models were observed the optimum dosage ,the rate of forming tumor , and tumor weight .Objective is to investigate the effect of cinobufacini on H22 Hepatocarcinoma transplanted in mice and explore the mechanism of cinobufacini. Methods:H22 Hepatocarcinoma were transplanted in mice.The mice were divided into five groups.Transplantation S180 mice tumor models were randomly divided into 5 groups. Investigate the anti-tumor effect of cinobutacini by the systematic observation on the morphologic tumor tissues .After building the model mouse , cinobutacini inhibitory rate on tumor ,thymus gland index ,spleen index and tumor tissues were observed by the optical microscope and the electron microscope .The inbred line C57BL/6 mice bearing Lewis lung carcinoma model is an ideal animal model used in the experimental study of lung carcinoma. Cinobutacini has the restraining effect on the function of the tumor cell .Cinobutacini has strong inhibiting effects on tumors. There is good applied foreground, should carry on a thorough research to it.MTT assay was used to examine inhibitions of Cinobutacini on growth of Hela ,SMMC-7721 and SGC-7901 cells .We aimed to analyze the inhibitory effects of cinobutacini on the proliferation and apoptosis of hepatoma cells or and SGC and Hela in vitro. Methods: human hepatic carcinoma SMMC-7721,human cervical carcinoma Hela , human gastric carcinoma SGC-7901 was cultured .it was treated with cinobutacini 0.0017,0.0068,0.0272,0.1088 and 0.4352μg/ml ,respectively .each concentration group was subdivided into four groups :treatment for 24,48,72,and 96h.the cells were harvested daily for four days .growth inhibition of the cell was measured by MTT assay .In vivo ,the tumor growth of mice with H22 was inhibited compared with the control group after animals were treated with cinobutacini at the three doses were respectively 20.92%,41.96% and 51.11%.The tumor growth of mice with S180 was inhibited also ,and the average tumor inhibition rates of it at the three doses was 21.23%,40.41% and 57.53%.after C57BL/6 mice ,the average Lewis lung tumor inhibition rates of cinobutacini at the three doses were respectively 17.65%,41.18% and 52.88%.Cinobutacini had significantly inhibitory effect on growth of human gastric carcinoma SGC-7901 cells, after 24,48,72,96 hours, the rate of cells growth suppression was 19.90%,28.78%,33.05% and 54.35% optics microscope, it can also be observed that evident morphological changes are found on the different tumor cells. The morphological changes of human gastric carcinoma SGC-7901 cells were observed after treated with cinobutacini for 24h,such as rounder, smaller, desquamation, sparse arrangement,vacuole of different size and etc. This paper adopted visible light electron microscopy to observe the cellular morphological features were observed these apparatus,such as:the number of cells decreased ,the cell shrunk ,the volume reduced ,the shape of cell nucleuses appeared like piece. We dealt with target cells at different times and concentrations. MTT assay was performed to test the inhibitory effect of cinobutacini on tumor cell lines.Distinct inhibitory effects were observed for low dosage groups. Neither in 48h nor 72h exposure, for high dosage group, and the inhibiting effects was very distinct and was grade with dosages. Under inverted microscope, it can also be observed that evident morphological changes are found on the different tumor cells.Results : compared with control group , cinobutacini has significantly inhibitory effect on human SMMC-7721 cells, SGC-7901 cell and Hela cell with concentration-dependent way. Cinobutacini could inhibit proliferation of human gastric carcinoma cells in time-dose-dependent response . Cinobutacini could inhibit the growth of various human tumor lines in vitro.