Experimental Research Of Anti-cancer Effects And Safety Of INj-alkaloids In Toad Skin

Tumor is one 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 less than that of cardiopathy. This makes it the second killer of human health.The alkaloids extracted from toad skin are broadly used as one of anti-tumor drugs at present. INj-alkaloids in toad skin is a safe and effective medicine to treat the PLC followed the metastatic tumer of lung .The research of pharmaceutical included pharmaceutical technology and quality standard. The main purpose of this article mainly works over the new Medicine prescription, manufacture technology, quality standard and pharmacology according to the request of the National New Drug Examination and Approval Method.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. 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, INj-alkaloids in toad skin quality standard was established, INj-alkaloids in toad skin is steady.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 content of one was determined by HPLC. 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.And anti-tumor efficacy and safety of INj-alkaloids in toad skin by acute-term toxicity test has been researched , in vitro pharmacodynamics test, safety test, pharmacology test and long-term toxicity test and so on.Through to acute-term toxicity test and LD50 determination, give the result of clinical medication, determiones the dose of medicine. About 2min's the drug poisoning mouse starts to appear the active reduction, peak wool, shuts item, walk haltingly, waggle, jitter. The mouse which are poisoned of large dose have a deep and slow breath, weak and limp. Beyond die the body-righting reflex of dead animals doesn't disappear, the breath from slow to tenuity. Dead mouse have respiration ceases first and cardiac arrest last, which are dissected have no abnormality acquire of organs. The undead ones who are killed 2 weeks later also have no abnormality acquire of organs.The in vitro experiment of pharmacodynamics shows alkaloids of toad skin's inhibition on tumor cells cultured in vitro using MTT analysis, and tumor control rate via cell appearance observed by inverted microscope. The results indicate that INj-alkaloids in toad skin inhibits evidently the human tumor cells proliferation in vitro and the inhibition depends a certain dose.Estimate the safety of the medicine, results of hepersensibility test, heamolyticus test and BV stimulus test on INj-alkaloids in toad skin show that INj-alkaloids in toad skin has no allergize on guinea pig, no obviously effect of in vitro heamolyticus and agglomeration in rabbit's red cell, it also has no obviously stimulation on rabbit's BV and can not cause pathological change of tissues of BV surrounding.The Pharmacology research uses to appraise the INj-alkaloids in toad skin regarding the animal cardiovascular, the respiratory and the nervous system influence. Through pressure transducer, tension transducer, needle-electrode connectting BL-420E biological function experiment system, determine INj-alkaloids in toad skin to take the different time compartment dogs femoral artery blood pressure, breath and the electrocardiogram around separately: observes the medicine with multi-purpose mouse activity recording instrument to the mouse spontaneous activity influence; observes the medicine with Fall-tiltboard method to the mouse coordination exercise influence; recording medicine to mouse pentobarbital sodium hypnosis function influence in subthreshold dose by Righting reflex deprivation. The results show that the anaesthesial dogs' systolic pressure reduce slightly, but there has not seen the remarkable difference about heart rate, diastic and the respiratory system.Moreover, INj-alkaloids in toad skin which in each dose has no obviously affects to the mouse coordnation exercise, automatic activities and pentobarbital sodium hypnosis function in subthreshold dose. The results indicate that INj-alkaloids in toad skin has certainly reduces the systolic pressure function which depends dose to the dog and it has no obviously affects regarding dogs' respiratory system and mouse's nervous system.Results of long-term toxicity test indicate that INj-alkaloids in toad skin has no effect on animal general activity, weight increased, organs' coefficient, hemogram and function of liver and kidney; every organ has no change of pathohistology. Stop to using this medicine, organs' coefficient, hemogram and function of liver and kidney of rats also have not been affected, so it is safe on the clinical application.