Basal medium and cells was determined by PARP fluorescence measurement, directly or after lysis. Three independent Independent experiments were performed, each performed in triplicate. All experimental metastases in vivo animal experiments were in accordance with German law for animal welfare, the reason Tze the protection of laboratory animals and with the consent of the local container Gestures performed. The adult female NMRI: nu / nu use M were obtained from Taconic Europe. The animals were housed in pathogen-free conditions in individually ventilated K Provisional housed under standardized conditions of the environment. They were feed and bedding and autoclaved drinking water ad libitum anges Uerten given. The breast cancer cells were transplanted into the brain MT3 and Topoisomerase subcutaneously into the left flank of each mouse. The animals were Feeder Llig in experimental groups with eight M Mice assigned.
Intravenously Se treatment with liposomes, the MTO or MTO, free, each p38 MAPK Signaling Pathway at a dose of 4 mg / kg was performed on day 3, 7 and 10. Control aids Mice were again Underground saline Solution in the same calendar year. Tumor diameter of the tumor was measured C s, w twice Weekly with a caliper. Tumor volume was cozy the L calculated length V ewidthT2 second In calculating the relative tumor volume of sc tumors, tumor volumes for each day of measurement on the first day of treatment. The K Body weight was determined twice a week. Mice were get Tet, as the M controls Mice Groups Showed the first signs of extracerebral tumor cell injection instead. Brains were isolated, snap frozen sections were prepared and cryo. The liquid surface The size E of the tumor was determined after cresyl violet F Staining followed by microscopic identification and calculated with the IQ EasyMeasure. The data were obtained from two independent experiments ngigen and are presented as mean values SD pharmacokinetic parameters using sc and intracerebral tumor-bearing female NMRI were added: Nozzles nu / nu-M. MT M Mice were treated with 3 with a single dose of 5 mg MTO / kg iv L Solution or encapsulated in liposomes or Lfluid Lfluid LG and Capecitabine randomized into groups of three M Mice. Referring to the timing of blood after the default to Anesthesiology iso isoflurane from the retro-orbital sinus before the Mice Were tet get a broken neck. Tumors and various organs were isolated and snap frozen until analysis.
Ammonia and S acid formate hexansulfonic, quilibriert with formic acid to pH 2: The samples were as a result of Johnson and quantification was by HPLC analysis using a C18-S column NUCLEOSIL, processes carried out with acetonitrile, 7, such as isocratic phase. MTO was detected at 610 nm and the concentration was calculated from a standard curve ofMTO. Each result represents the maximum mean of three mice M Determined twice. Pharmacokinetic data were closing Lich calculated using the program Win NONLIN from Scientific Consulting Inc., Apex, North Carolina, USA. The pharmacokinetic parameters for the distribution of the plasma on the basis of a two-compartment model for an iv bolus injection calculated. Statistical analysis All data are expressed as standard deviation of the mean. Statistical comparisons of the in vitro data were performed with the unpaired Student t-test two populations, w During the statistical analysis of the in vivo data was performed using the U test of Mann and Whitney with.