Design, Synthesis And Biological Evaluation Of 10,11-Methylenedioxy-camptothecin Derivatives And Phosphate Ester Prodrugs Of Fulvestrant

Structure modifications to known active molecules had became an important way for drug development. This dissertation focuses on a class of molecules targeting and hormone anticancer drugs in order to find higher activity, less side effects, superior pharmacokinetic antitumor candidate compounds through structural modifications. Series of compounds were designed and synthesized. The structures of the synthesized compounds were all characterized and biological activities evaluations were well done. The content of this dissertation contains two parts which were as follows:1. Design, synthesis and biological evaluation of 10,11-methylenedioxy-camptothecin derivatives as survivin inhibitorsCamptothecin derivatives (CPTs) has been regarded as one of the three great anticancer drugs in 1990s, displaying broad spectrum anticancer activity and of great research and application value.10,11-methylenedioxy camptothecin (FLU 8) is a novel small molecule inhibitor recently identified by Fengzhi Li. Even though FL118 is structurally similar to irinotecan, it exerts its anti-cancer effect by inhibiting survivin gene expression instead of inhibiting DNA Top-I. Although FL118 has been demonstrated with superior and broad antitumor efficacy, we hope that through studies of the structure-activity relationship (SAR) between FL118 and its closely structure-related analogs, we are able to obtain overall better small chemical molecules that are highly relevant to the FL118 structure but show improved therapeutic index without increasing adverse toxicity to normal tissues.The key intermediate ring CDE can be synthesized by 15 steps from acetone and ethyl oxalate including condensation, substitution, rearrangement, cyclization and chiral resolution etc. Different ring A can be synthesized by 2-4 steps from 3,4-methylenedioxybenzaldehyde,3,4-methylenedioxyacetophenone and 3,4-methyl-enedioxyaniline by nitration, reduction and condensation. After connecting the two parts of intermediates and further structural modifications.42 derivates of 10,11-met-hylenedioxy-camptothecin and 28 important intermediates were synthesized, which were firstly reported. We optimized the synthetic route of 10,11-methylenedioxy-cam-ptothecin and improved the chiral separation method of tricyclic intermediate. The total yield was improved to 5.7% from 2.1%.Then, the anti-cancer activities of compounds were tested systematically in vitro and in vivo. The compounds ZJY-7-6, ZJY-7-17 showed great inhibition of survivin gene expression in human ovary cancer cells (2008) at the concentration of 0.1 μ.M. The compound ZJY-7-6 was cytoxic to human hypopharyngeal carcinoma cells (FaDu) even at the concentration of 1nM in vitro. There were six compounds showed great cytotoxicity to human lung adenocarcinoma cells (A549), and the half maximal inhibitory concentration (IC50) values were 1-10 nM. The IC50 values of five compounds were 10-50 nM. There were fourteen compounds showed good cytotoxicity to human leukemia cells (K562), and the IC50 values were 0.1-1 μM. At the xenografts model of FaDu and colon cancer cells (SW620), the maximum tumor growth inhibitions (MTGI) of compounds ZJY-7-5, ZJY-7-14, ZJY-7-15 and ZJY-7-17 were more than 90% after the administration for 10 days. In addition, the body weight loss of mice for compounds ZJY-7-15 and ZJY-7-17 was less than 15%, which have the potential for further develop. Preliminary study of structrue-activity relationship (SAR) indicated that the antitumor activity was best when the 7-position of 10,11-methylenedioxy-camptothecin was substituted by lipophilic groups. The antitumor activities were lost when 20-hydroxy was esterified by substituted benzoic acids. The inhibitory activity against A549 cells gradually weakened with the extend of connecting arm of glucose. The inhibitory activity against A549 cells was higher than K562 cells in both 7-position modified derivatives and 20-position modified derivatives.2. Design, synthesis, and biological evaluation of 1,3-cyclic propanyl phosphate ester prodrugs of fulvestrantFulvestrant is a selective estrogen receptor antagonist, which is used for the treatment of hormone receptor-positive advanced breast cancer in postmenopausal women with disease progression following antiestrogen therapy. To improve the poor stability, low oral bioavailability, rapid metabolism and short half-life time of fulvestrant, we connect the 3-phenolic hydroxyl of fulvestrant with cyclic phosphates of different substitutions, and obtained 9 firstly reported cyclic phosphate prodrugs of fulvestrant.Three compunds were selected for stability test, fulvestrant was chosen as negative control.We studied the effect of storage temperature and time on the purity of compounds, and the results demonstrated that the stability of cyclic phosphate prodrugs were improved significantly compared with fulvestrant.We conducted the in vitro release experiments of fulvestrant cyclic phosphate prodrugs. the results indicated that the prodrugs can gradually release fulvestrant under the catalyzed of enzyme. The metabolic rate was more than 80%. The prodrugs can be metabolized completely and released the original drug.