Neovascularization Inhibitor 2-me, And Coumarin Skeleton Analogues And Biological Activity

The growth and metastasis of tumor cells depend on the tumor angiogenesis, which supply them the necessary nutrients and oxygen. Hence, anti-angiogenesis agents have became a new target of research on anti-tumor drugs. 2-Methoxyestradiol (2-ME, 1), with anti-angiogenic activity, is under developing as an new anti-cancer drug candidate in phaseâ… -â…¢clinical trials for treatment of various solid cancers, especially breast cancer, prostate cancer and multiple myeloma. Based on the structure character of 2-ME, a series of its non-steroid analogs were designed to explore their inhibitory activity of angiogenesis.This dissertation includes four parts:1) The synthesis of control compound 2-ME and the Improvement of its synthetic methodThe 6 steps among the 7-step synthetic route of 2-ME have been improved: shortened one steps, simplified the operation, and increased the overall yield of improved steps by about 12% and made it more suitable for scale manufacturing.2) Design and synthesis of the target compound 3-p-hydroxyphenyl-4 -methyl-6-methoxy-7-hydroxycoumarin (6), as a nonsteroidal analogue of2-MEWe designed 3-p-hydroxyphenyl-4-methyl-6-methoxy-7-hydroxycoumarin (6), a nonsteroidal analogue of 2-ME. It contained the necessary pharmacophore of 2-ME and a 3- phenylcoumarin skeleton instead of steroid scafford. After the effort on 5 different synthetic routes, we eventually accomplished the synthesis of (6).3) The structure modification of 4-position of 3-p-hydroxyphenyl-4-methyl -6-methoxy-7-hydroxycoumarin (6)The structural modification of 4-position of compound (6) was carried out via two ways: firstly building coumarin skeleton then modifying group at 4-position, or pre-functioned at 4-position then building coumarin skeleton. A series of 4-methyl, 4-hydroxyolmethyl,and 4-aryl coumarine derivates were synthesized and some unexpected behavior reactions were discussed. Another series of 2-ME analogs, 2-aryl-6-hydroxyl-5-methoxyl-benzofuran-3-ones, have also been synthesized as anti-angiogenesis candidates.4) Bio-activity tests and resultsThe target compounds and some intermediates are tested against the growth of human umbilical vein endothelial cells (HUVEC) and the expression of vascular endothelial growth factor (VEGF). Most of them showed potent inhibitory activities. The target compound (6) exhibited better inhibitory activities against the growth of HUVEC (EC₅₀ = 5.69μMol/L, IC₅₀ = 256.12μMol/L,TI=45.01) than 2-ME (EC₅₀= 8.95μMol/L, IC₅₀= 70.87μMol/L,TI = 8.25) as positive control compound. The nonsteroidal analogue of 2-ME, 3-p-hydroxyphenyl-4-methyl-6-methoxy-7-hydroxycoumarin (6) and its analogues showed good inhibitory activities in the bioassay, and this provided a clue for the research on new types of neovascularization inhibitory agents.