Design,Synthesis And Biological Evaluation Of Novel Indole And Isoindole Derivatives As Tubulin Polymerizaiton Inhibitors

Microtubules are important structural components of eukaryotic cell and they can assemble into a variety of structures with other proteins,like spindles,centrioles,flagella and so on.Based on these structures,microtubules have lots of biological functions.For example,It is a very important function that microtubules can participate in mitosis as spindles.Cancer cells can't divide rapidly without the function of spindles.If spindles are inhibited,the mitosis arrest will occur and then the growth of tumor tissue will be controlled or introduction of tumor apoptosis will happen.In addition,microtubules have vital 'effect on keeping cytoskeleton shape.Tumor vascular endothelial cells are not mature enough to keep their shape where they depend on cytoskeleton network.Destorying the cytoskeleton network will lead to the collapse of vascular endothelium cells and then cause tumor necrosis.Therefore,microtubules become a favored target in anticancer drug development.After the identification of tubulin as potential target for anticancer drugs,extensive research about tubulin inhibitors has been observed in a dozen years.Natural products and chemical synthetic compounds are discovered frequently.Analogues of the natural products have been synthesized and their activities are compared with lead compounds.Chemical synthetic compounds mainly mimick the natural products that have prominent activity.The rapid development of organic chemistry leads to extensive research where new tubulin inhibitors with strong activity are synthesized.Among them,heterocyclic compounds occupy largely and are studied extensively.Because of their rich electron and modifiable groups,heterocyclic compounds can be designed into a variety of compounds.According to the mentions above,my research can be divided into four parts:1.Comprehensively and systematically summarize the structure,anti-cancer activity and pharmacological mechanism of the tubulin inhibitors reported in recent years.Through searching literature,there are many reports studying indole,but the reports about pyrimidine are a few.The research about primidine-indole tubulin inhibitors is fewer.The novel tubulin inhibitors A mentioned in 2011 draws our attention.The structure of A belongs to primidine-indole tubulin inhibitors.We connect the pyrimidine fragment in compound A and another novel compound D-64131 with an amino group according to the combination of active structural moieties theory.Finally,thirty-one novel pyrimidine-indole compounds ? and ? are achieved.All compounds are characterized by proton magnetic resonance spectroscopy(1H NMR),nuclear magnetic resonance spectroscopy of carbon(13C NMR),El low-resolution mass spectrometry(MS),elemental analyzer(EA)and their specific structure types are summarized as follows:I a I s:2-(4-methyl-piperazin-l-yl)-N4-substituted-indol-5-nitro-pyrimidin-4-yl amino.the synthesized compounds have reached nineteen.? a-?1:2-morpholin-N4-substituted-indol-5-nitro-pyrimidin-4-ylamino,the synthesized compounds have reached twelve.2.In previous study,we have discovered compound B that have strong anticancer activity as tubulin inhibitors.In order to study structure-activity relationship of this compound,we modify the substituted groups on phenyl and phenyl itself.Finally,ten isoindole-1,3-dione derivatives(Compounds ?)are achieved.The specific structure types are showed as follows:?a-?j:2-(3-substituted-5-(3,4,5-trimethoxy-phenyl)-(1,2,4)triazol-4-yl)-isoin dole-1,3-dione,synthesized compounds have reached ten.3.Biological evaluationsWe adopted MTT assay to test the bioactivity in vitro.Only a part of compounds were tested.We selected four human cancer cell line to do experiment,including human colon cancer cell line(HT-29),human breast cancer cell line(MCF-7),human lung cancer cell line(A549),human breast cancer cell line(MDA-MB-231).After the test,some compounds with high activity were discovered.Comparing with CA-4,most of the target compounds showed inhibition to MDA-MB-231.Among them,Compound I i and I s showed IC50<10?M in MDA-MB-231.Especially,Compound I s displayed the most strong activity in all target compounds and inhibited HT-29,A549,MCF-7,MDA-MB-231 cancer cell line with IC50 values of 5.56,14.36,13.93,5.01 ?M.It was thought to have potential development value.In addition,we tested the inhibition ability to tubulin for some synthesized compounds and they all served as inhibitors of tubulin polymerization in some degree.Compound I b,I e,I s displayed a large inhibitory.activity against tubulin polymerization(IC50 = 15.2,19.2,11.2 ?M for inhibition of tubulin polymerization).Compared with vinorelbine(IC50 = 2.0 ?M),their inhibitory activity were close to it.Next plan,we prepared to modify the structures in order to get the compounds with higher activity.4.Molecular dockingCompound I s was docked with tubulin because of its strongest activity in all compounds and the binding mode was confirmed between compound I s and colchicine binding site...