Design, Synthesis And Biological Evaluation Of DPP4and EGFR Inhibitors For Treatment Of T2DM And NSCLC

Based on traditional medicinal chemistry, computer-aided drug design and modern synthetic technology, the dissertation for Ph.D. degree focused on design, synthesis and biological evaluation of Dipeptidyl peptidase IV inhibitors and Epidermal Growth Factor Receptor inhibitors and the methodology of Au(I)/Ag(I)-catalyzed cascade approach for the synthesis of polycyclic heterocyclic compounds.Firstly, based on previous work in our group, we developed the work for the design, synthesis and biological evaluation of a-amino and (3-amino pyrrolidine-2-carbonitrile derivetives as DPP4inhibitors. Based on analysis of metabolites of lead compound, the rational design was carried out followed by introduction of different substituted group, displacement of aromatic ring with aromatic heterocyclic, and prolonging the side chains of lead compound. And47compounds (A1-A22and B1-B25) were designed, synthesized and evaluated to inhibit DPP4and other serine protein family activities. Among these compounds, the DPP4inhibitory activities of11compounds were less than20nM, the DPP4inhibitory activities of8compounds (A1, A3, A7, A8, A9, A14, A15and A17) were less than10nM. The IC50values were4.1nM,2.7nM,6.2nM,4.6nM,5.7nM,10.0nM,5.0nM and4.0nM, respectively. Moreover, most compounds showed good selectivity. Considering to DPP4inhibitory activities and selectivity in vitro, compounds A14, A17, B12and B25were selected for acute efficacy evaluation by the oral glucose tolerance test (OGTT) in ICR mice, and the results showed that compounds significantly reduced the blood glucose level after glucose challenge, and showed in dose-dependent manner. Moreover, compounds A17and B25were selected for the chronic effects experiment, and the results showed these two compounds increased the basal plasma insulin content significantly. Besides, compounds A14, A17and B25showed good pharmacokinetic in SD rats, and the oral bioavailability was16.8%、37.8%and22.8%, respectively. Besides, compound A17was selected for metabolite analysis in SD rats. Moreover, compounds A14, A17, B12and B25did not block hERG channel and showed no liver metabolic enzymes P450inhibition, such as CYP2C9, and compound B25showed good safety in acute toxicity study in ICR In the research of design, synthesis and biological evaluation of EGFR inhibitors for treatment of non-small cell lunch cancer. Based on the bio-isosterism and pharmacophore combination principles,56compounds (C1-C30and D1-D26) were designed, synthesized and evaluated for inhibition of EGFR wt and EGFR T790M/L858R. Among these compounds, the EGFR wt inhibitory activities of14compounds were less than20nM, the EGFR wt inhibitory activities of13compounds were less than10nM, and the EGFR wt inhibitory activities of5compounds were less than5nM. Moreover, the EGFR T790M/L858R inhibitory activities of10compounds were less than10nM. Considering to the inhibitory activities of EGFR wt and EGFR T790M/L858R in vitro and anti-proliferative activities in A431and NCI-H1975cell lines, compounds C4, C18, D14, D15, D17and D22were selected for further study to attenuate EGFR activity and EGFR downstream signaling molecules in A431and NCI-H1975cancer cells. The results showed that compounds C4, D14, D15, D17and D22at0.1μmol/L almost completely blocked the autophosphorylation of EGFR wt in A431cells, and compounds D14, D15, D17and D22dose-dependently blocked EGFR T790M/L858R phosphorylation and downstream signaling in NCI-H1975cells. Moreover, compound D15was tested the activities on other members of the EGFR family ErbB2, ErbB4, as well as11types of tyrosine kinases using an in vitro kinase assay. These data demonstrated that compound D15was selective inhibitor of members of the EGFR family.Polycyclic heterocyclic compounds had an important role in medicinal chemistry. In the methodology, we developed an efficient and facile Au(Ⅰ)/Ag(1)-catalyzed cascade method for one-pot synthesis of the complex polycyclic heterocycles through treatment of two simple starting materials. The strategy features Au(I)/Ag(I)-catalyzed one-pot cascade process involved in the formation of three new C-N bonds in high yields, and with broad substrate scope. Moreover, the structure of compound3Aa was characterized using X-ray crystallography.In summary, based on traditional medicinal chemistry, computer-aided drug design and modern synthetic technology,103compounds (A1-A22, B1-B25, C1-C30and D1-D26) were designed and synthesized for evaluation of DPP4and EGFR. In the research of DPP4inhibitors, compounds A14, A17, B12and B25showed significantly reduced the blood glucose level after glucose challenge in ICR mice, and compounds A17and B25increased the basal plasma insulin content significantly in KBS dbldb mice. Moreover, compounds A14, A17and B25showed good pharmacokinetic in SD rats. And compounds A14, A17and B25did not block hERG channel and showed no liver metabolic enzymes P450inhibition, such as CYP2C9, and compound B25showed good safety in acute toxicity study in ICR mice. In the research of EGFR inhibitors, compounds C4, D14, D15, D17and D22at0.1μmol/L almost completely blocked the autophosphorylation of EGFR wt in A431cells, and compounds D14, D15, D17and D22dose-dependently blocked EGFR T790M/L858R phosphorylation and downstream signaling in NCI-H1975cells. Moreover, compound D15was a selective inhibitor of members of the EGFR family. In addition, Au-catalyzed cascade approach for synthesis of polycyclic heterocyclic compounds was developed. All the discussed works will be a good foundation for the developing new drugs with independent intellectual property rights.