Antiproliferative effect and mechanism of nolatrexed dihydrochloride
|School||First Military Medical University|
|Keywords||Nolatrexed hydrochloride Antitumor In vitro In vivo Distributed Apoptosis Mechanism of action|
The purpose hydrochloric nolatrexed new classic thymidylate synthase inhibitor, is a new generation of anticancer drugs. Study abroad for its anti-tumor spectrum screening is not yet complete, in charge only of its in vivo pharmacodynamic study L5178/TK - sup> and transplanted human cervical cancer mice, found no drugs in the organization distribution of information, and its mechanism of action is limited to the inhibition of thymidylate synthase, many of the more important information on the application of the drug is still not perfect. The main purpose of this experiment is to: (1). Understanding of hydrochloric acid nolatrexed cultured in vitro inhibition of tumor cell proliferation, and screening for drug-sensitive tumor cell lines; (2). The visits hydrochloric nolatrexed charge SRS-82 ascites tumor / solid tumors as well as Dutch S-180 ascites tumor / solid tumor efficacy: (3). Hydrochloride single nolatrexed, given orally 100mg.kg -1 sup> mice bearing solid drug in the blood, heart, liver, spleen, lung, kidney, brain and tumor tissue organization parts of the distribution; (4). From apoptosis angle nolatrexed hydrochloride possible mechanisms of action, and apoptosis in the speed and the relationship between intensity and drug effects of time and drug concentration levels for analysis; (5). Research hydrochloric nolatrexed TS enzyme mRNA levels, to explore the drug whether there is inhibition of the cascade enlarged. The method according to the purpose of the experiment using the following experimental methods: (1) 10 kinds of tumor cell lines in vitro culture of non-selective and normal cells were added to a series of concentrations of hydrochloric acid nolatrexed cultured for 72h by MTT assay. detection of drugs on the proliferation of tumor cell lines and normal cells, calculated drugs on the cells of the IC 50 value; (2) Netherlands SRS-82 ascites tumor / solid tumors and Dutch S-180 ascites tumor / tumor-bearing animal models of solid tumors in mice, respectively 100mg.kg -1 sup> 75mg.kg -1 sup> 50mg.kg -1 sup> dose continuous administration 7d, 1 / d, 0.9% NaCl solution as a blank control, 5-FU10mg.kg -1 sup> as a positive control of ABN ascites tumor-bearing mice prolong survival time as an indicator, indicator of the rate of increase slows down tumor weight of mice bearing solid, the visits hydrochloric nolatrexed efficacy of tumor-bearing mice. (3). Single dose administration 100mg.kg -1 sup>, respectively, after the administration 0,5,45,225,300 and 345min, blood, heart, liver, spleen, lung, kidney, brain and tumor tissue after the crushing of the organization, plus acetonitrile protein precipitation, centrifugation supernatant dry vacuum dryer, mobile phase, detection at 257nm wavelength at each sample drugs by high performance liquid chromatography The concentration of hydrochloric acid nolatrexed distribution charge SRS-82 solid tumors in mice. (4). The process of by hydrochloric nolatrexed, LoVo colorectal cancer cells, DNA fragmentation analysis, as well as ordinary light microscope and transmission electron microscope for cell morphology observed apoptosis, flow cytometry analysis of drug concentrations level and duration of action on tumor cell apoptosis intensity marrow degrees. (3lfeg by hydrochloric nolatrexed treatment and control groups iodine [cultured the SRS82 tumor cell lines IWA, UV spectrophotometer total RNA content, the same amount of total IUqA primer and R labeled the RTM gel analysis system the B enzyme WA level changes. Results (l). tumor cells K562 of hydrochloric acid nolatrexed of in vitro culture, NSI, H Shen Yu, Mp, o, nVb, Q uality Wang, SRS cut places places Bo other normal cells W93, CHO and h 55.8, knife-l, 213,16.9,575, OD12, 952,10 two .019,813 and 325uM, consistent with China's Xi Shu reported data; K early to 575,1876. ZdesuM hydrochloric Lola song grams selective inhibitory effect on normal cells and tumor cells. (2 salt jumped pull Qu grams of cis SRSy ascites rot real ugly bowel S-180 ascites rot real Peng mice have extended survival time or less? role of tumor tissue, a significant difference with the blank control group; Dutch ascites tumor in mice to better is Kazakhstan slightly administration efficacy; solid tumors in mice, efficacy acid drug after the dose is increased and strengthened. (). hydrochloric nolatrexed the lha In oral administration Netherlands SRS-82 solid tumors in mice, the drug absorption relentless is slow, but eliminate leg, identified in the blood of the highest attainable sway from side to side for 12 snakes ice heart, liver, spleen, lung, kidney, brain boron? reached the highest concentration levels in the tumor tissue were 157 o.l, 11.5,177,58 *, 161, l Xiaosi hasty. (4 body eve of training IbVt3 iii cancer cells make by hydrochloric nolatrexed, processing, confirmed by agarose gel electrophoresis of DNA fragmentation analysis and light microscopy Wo D electron microscopy Morphological observation cited gate death hydrochloric nolatrexed's role helm u one. endowed cell vessels cytometry analysis showed that lbVb cell lines after drug treatment 480mn, the number of apoptotic cells in the highest; apoptosis drug concentration exceeds a certain concentration, drug-induced apoptosis strength no longer increases. (The experimental design primers synthesized can be specifically amplified h enzyme DDNA in mice, the length of amplified product were 48p and 570be; cultured in vitro the SRS82 cell lines via 20,10, i 125,0. a 'hydrochloride Luo pull Qu grams at e extracted the same amount of total RNA for the RTM PCR product was compared with the control group, respectively, by approximately knife%, 6h, 46%, 4i, 44%, its the B enzyme InRNA level with the concentration of the drug liter high decline. Conclusions (1) the of hydrochloric acid nolatrexed of in vitro culture K56 the NSI, Hp, Mn, h, lbVb, QSG7703, SRSy, 5180 \\ _T24 tumor cells have significant anti-proliferative effects, including field forbearance The silent and h cell lines are highly sensitive hydrochloride nolatrexed,. of hydrochloric nolatrexed of normal cell growth inhibition, but the effect was weak, selective drug action. (2 hydrochloric acid nolatrexed of the Netherlands - 3-alm ascites to withdraw solid tumors and a of Vitex Pán field ascites rot solid tumors were significantly inhibiting taboo u role of mouse ascites tumor efficacy, to have been prepared Kuang administration best efficacy with real myeloid to drug dosage increase its anti-tumor Mindanao enhanced the four types? tumors, its efficacy and lug'M; the 'ry administration group efficacy comparable or better. Q) Holland the SI (SS solid tumors to mice times were given nolatrexed hydrochloride Ind drugs in organs and tumor tissue required a longer time to peak, but eliminates the rapid distribution of the drug in an organ, tissue peak concentration values ??from highest to lowest: kidney>?