Design,Synthesis,Biological,Evalution And Structure-activity Relationship Of The Anti-cancer Agents Taragents Targeting MMP-2

Malignant tumor is one of the diseases that seriously threaten peoples' health,being the second most commonly diagnosed cancer in the tumor patient of the world.Tumor cell migration and invasion is one of the most important events contributing to tumor dissemination,and its prevention may arrest malignant evolution.Tumor invasion is a complex,multistep process and many of the stages require degradation of the extracellular matrix.Matrix metalloproteinases are a family of extracellular zinc-dependent neutral endopeptidases collectively capable of degrading essentially all ECM components.Under normal physiological conditions,MMPs not only play important roles in physiologic ECM remodeling,but also are involved in pathological conditions.There is also considerable evidence indicating that some MMPs play important roles in tumour invasion and metastasis.MMPs as a therapeutic target offered opportunities for anti-cancer drug development.The primary aim of research into the mechanisms of tumor invasion and metastasis formation is to identify new strategies for more effective therapy against this most deadly aspect of human cancer.Recently,matrix metalloproteinases inhibitors have attracted great interest in the drug discovery community.In this thesis,we focus on the identification and modification of novel molecular probes that interfere with the binding region of MMP-2 as a demonstration of using highly specific and selective small molecule agents to inhibit MMP-2.Means of Computer Aied Drug Design was taken,in which,combined with Docking calculation;parative Molecular Field Analysis were applied in the Three Dimensional QuantitativeStructure and Activity Relationship study of known matrix metalloproteinases inhibitors.Virtual screening program and in vitro anti-cancers and MMP-2 inhibitory activities evaluation were introduced for initial screening.Then candidate five compounds containing benzenesulfonate sulfonamide and skeleton derivatives Hit 1(IC50 MMP-2=62.60?M),dihydropyrazole sulfonamide skeleton derivatives Hit 3(IC50 MMP-2=3.84 ?M),Hit 6(IC50 MMP-2=16.81 ?M)Hit 8(IC50 MMP-2=1.31 ?M)and dihydropyrazole thiazole skeleton derivatives Hit 10(IC50 MMP-2=11.72?M)were obtained and selected for further optimization and pharmacophore studies.Based on the structure of lead-compound and SAR(Structure-activity Relationship)of synthesized analogs,130 pyrazoline derivatives have been designed(128 of them were firstly reported),synthesized,characterized by NMR,MS and ESI,among which 6 compounds were provided with single crystal X-ray structural analysis.Many of the which compounds have good anti-cancer activity.The results were concluded as follows:(1)A series of novel MMP-2 inhibitors(A13?A39)containing phenyl benzenesulfonate sulfonamide skeleton has been synthesized and tested their biological activity.These compounds exhibited potential inhibitory activity on MMP-2 and anti-proliferative activity on Hela.Compounds A22 exhibited the best inhibitory activity on MMP-2 and Hela with IC50 values of 0.38 and 0.63 ?M,respectively.In order to further investigate the mode of action between them,we performed molecular simulation.The result showed that there were few interactions between the amino acid residues in the Batimastat action sites of MMP-2 with the compound,which might play an important role in the activity.(2)According to the skeleton of dihydropyrazole sulfonamide derivatives Hit 3(IC50=3.84?M),Hit 6(IC50=16.81?M),Hit 8(IC50=1.31 ?M),further structural modification was performed,eighty-seven dihydropyrazole sulfonamide derivatives were synthesized,and eighty-five of them were new compounds.Furthermore,the crystal structure of compounds B25,C9,C17,and C23 were determined by single crystal X-ray diffraction analysis.Firstly,various substituents were introduced in C-4 position of the dihydropyrazole.Then structural modification was proceeded on the C-4 or C-5-phenyl ring of Hit 3 and Hit 6,altogether fifty-five phenoxy ring and naphthalen sulfonamide derivatives were obtained in this part.Among B1-B25,compound B3 exhibited the most potent inhibitory activity with an IC50 value of 0.33?M for MMP-2,compounds B13 showed anticancer activities against MCF-7 with the IC50 of 1.62.Besides,the rest compounds C1-C32 exhibited very good bioactivity.The most potent inhibitor against MMP-2 and A549 were compounds C11(IC50=0.21 ?M and 1.90?M,respectively.The 3D-QSAR models with reliable predictability were established to analyze the structure-activity relationship and provide pharmacophore clues for further structural optimization.The 3D-QSAR contour map suggested an increasing binding affinity might be obtained by replacing the C-4-phenyl ring with a slightly larger template which also can cause a lower electron density.Thus,benzodioxan motif was introduced in C-4 position and thirty 4-(5-(benzo[d]dioxol-5-yl)or 5-(2,3-dihydrobenzo[b][1,4]dioxin-6-yl)-containing 3-(4,5-dihydro-1H-pyrazol)benzenesulfonamide derivatives were designed as potential MMP-2 inhibitors and predicted to have a positive progress with a sound cancer therapeutic benefit.Finally,the most potent MMP-2 inhibitor and antitumor owing 3-(4,5-dihydro-1H-pyrazol)benzenesulfonamide D6(IC50 MMP-2=1.31?M)and D10(IC50 Hela=0.38?M),respectively,was identified.The introduction of 3-(4,5-dihydro-1 H-pyrazol)benzenesulfonamide structure reinforced the combination of our compounds and the receptor,resulting in progress of bioactivity.(3)All novel dihydropyrazole containing thiazole skeleton derivatives MMP-2 inhibitors had been synthesized and evaluated for their MMP-2 inhibitory and antiproliferation activities of the MCF-7,HepG2 and Hela cell lines.The results showed that compound E17 exhibited the most potent MMP-2 inhibitory activity with IC50 of 2.80?M,but compound E4 displayed the most potent activity against MCF-7 cell line(ICso=1.03?M).The SAR analysis result showed that an electron-donating group in the position R1 and R3 could improve the inhibitor activity.In addition,we have also carried out molecular docking between compounds E1-E18 and MMP-2 to reveal the possible combination between them.