Synthesis And Biological Evaluation Of Novel Water-soluble Cyclane-aminol10-hydroxycamptothecin Analogues

DNA Topoisomerase I (Topoisomerase I, Topo I) has a relationship with DNA replication, transcription, recombination, and repair and other processes. It is an important target for antitumor drugs. Camptothecin derivatives are currently the only anticancer drugs as DNA topoisomerase I inhibitors. It has clear mechanisms, strong anticancer activity, broad spectrum with no cross resistance and other advantages, but it still has vomiting diarrhea, and serious adverse reactions such as bone marrow suppression.10-hydroxycamptothecin (HCPT) was used as lead compound, cyclane-aminol was introduced into HCPT at C9to make3novel water-soluble cyclane-aminol HCPT analogues by Mannich reaction, such as10-hydroxyl-9-L-prolinol(+)methylcamptothecin (PRPT),10-hydroxyl-9-(4’-hydroxy)piperidinylmethylcamptothecin (PPPT) and10-hydroxy-9-(4’-hydroxyethyl)-piperazinylmethy camptothecin (QPPT), and got their oxidation9-methylene-10-carbonylcamptothecin. The main influencing factors in the Mannich reaction were investigated, designed the conditions of synthesis and purification based on related substances structural analysis by LC-MS.Cyclane-aminol HCPT analogues was evaluated with biological mothods. The ratio of lactone/carboxylate of PPPT and PRPT in the plasma was appropriate and better than TPT and HCPT. The solubility of PRPT in octanol and PBS was far superior to TPT and HCPT.The metabolites of cyclane-aminol HCPT analogues in rats by LC-MS were evaluated. Results showed that the major metabolic pathway was capturing oxygen free radicals in the cell with producing H2O2. The major metabolites of PRPT were sulfated metabolites, deoxidized metabolites, methylated metabolites, and prototype drug was28%. The major metabolites of PPPT were9-methylene-10-carbonyl-CPT and the prototype drug was80%. The major metabolites of QPPT were9-methylene-10-carbonyl-CPT with no prototype drug excretion. The main metabolites of TPT were prototype drug with little other metabolites. These results indicated that the metabolism of PPPT and PRPT was better than QPPT.The method for determination of intracellular H2O2was established by PF1selective fluorescent probe. When H2O2was in3.1-100μM concentration range, the curve linear was relationship with H2O2concentration. Results showed that PRPT and PPPT could enhance H2O2concentration in MCF-7and BGC-823tumor cells, induce apoptosis on tumor cells, do not induce apoptosis and death on PBMCs. In summary, the novel water-soluble cyclane-aminol10-hydroxycamptothecin analogues have advanced research results, the molecula mechanism have further study.