Design, Synthesis And Antitumor Activities Of Benzotriazines And Pyridotriazines

Angiogenesis is required for the growth and metastasis of solid tumors.RTKs are enzymes that can transfer a phosphate group from ATP to a tyrosine residue of proteins.The direct inhibition of the tyrosine kinase activity of VEGFR will result in the reduction of angiogenesis and the suppression of tumor growth.Several monoclonal antibodies and small-molecule inhibitors targeting tyrosine kinases have been launched as anticancer drugs. In this thesis,small-molecule inhibitors being developed were reviewed.A three-dimensional quantitative structure activity relationship（3D-QSAR） model of 4-amino-fluoro[2,3-d]-pyrimidines,which were reported as VEGFR inhibitors,was established by using the comparative molecular field analysis（CoMFA）.After the analysis of the results of the 3D-QS AR model and the ATP-binding model to tyrosine kinase,referring to the structures of vatalanib and vandetanib,1,2,3-benzotriazine and pyrido[3,2-d]-1,2,3-triazine were designed as the main structure of the target to replace 4-amino-fluoro[2,3-d]-pyrimidine.4-Substituted anilino-1,2,3-benzotriazine（A01-A26）, 7-alkoxy-6-methoxy-4-substituted anilino-1,2,3-benzotriazine（B01-B17,C01-C17,D01-D17, E01-E16）,and 4-substituted anilinopyrido[3,2-d]-1,2,3-triazine（F01-F16） were synthesized. The structures of all the target compounds were determined by application of infrared（IR） and nuclear magnetic resonance（NMR） spectra as well as mass spectra（MS）.Some characteristics about IR and ¹ H-NMR spectral data of the target compounds were summarized.In IR spectra,the character peaks of 1,2,3-benzotriazine were near 3100,1620, 1570,1500 cm^-1.The character peaks in IR spectra of pyrido[3,2-d]-1,2,3-triazine were about 1610,1570,1500 cm^-1.In ¹H-NMR spectra,the chemical shift of NH were displayed at 10.01-10.20 ppm,11.59-11.76 ppm,9.50-9.81 ppm and 10.36-10.70 for compounds A01-A17, A18-A26,B-E and F,respectivly.Protons at C5 and C8 of A01-A17 appeared as a doublet at about 8.20 ppm and 8.60 ppm,respectively.The ¹H-NMR spectra of A18-A26 indicated that H-5 and H-8 were at about 8.40 ppm and 8.80 ppm.After the introduction of alkoxy groups to C-6 and C-7（B,C,D,E）,protons at C5 and C8 both displayed as singlets at about 7.60 ppm and 7.90 ppm.The ¹H-NMR spectra of F series indicated that protons at C7 and C8 appeared as doublets at about 8.70 ppm and 8.20 ppm,respectively.The synthetic procedure of vatalanib succinate,a specific VEGFR inhibitor under phaseâ…¢clinical trials,was improved.Starting from phthalide and pyridine-4-carbaldehyde, Vatalanib succinate was prepared in five steps of reactions in an overall yield of 22.2%.The improved synthetic procedure has the advantage of low cost and is suitable for industrial production.The antiproliferative effects of these compounds on murine microvascular endothelial cells（MVEC） were determined compared with vatalanib succinate.Compounds B09,B10, B11,C08,E12,E15,E16 had similar activities with vatalanib succinate(GI₅₀ = 38.15μM). Compounds B01,B04,B05,B12,B13,C01,C03,C05,C07,C09,C10,C12,C13,C16,C17, D03,D05,D09,D10,D11,D12,D13,D15,D17,E01,E05,E09,E13,F01-F16 were more effective than vatalanib succinate.The antiproliferative activities of compounds C12,D17, F07,F09 in breast cancer T47D cells,prostate cancer DU-145 and PC-3,murine Lewis lung cancer LL/2 cells,murine melanoma B16F0 cells,and human umbilical vein endothelial cells （HUVECs） were determined using MTT assay.Compounds C12,F07 and F09 were about ten times more potent than vatalanib succinate.Compounds C12,F09 were docked into the ATP binding site of VEGFR-2 using Autodock 4.0 software.The binding models of compounds C12,F09 were similar to that of ATP.It was deduced that compounds C12,F09 might inhibit the activities of VEGFRs as an ATP competitive inhibitor.The action mechanism of the target compounds are under investigation.