| Malignant tumor is one of the most important diseases that threaten the human health in the twenty-first century, which has seriously endangered the lives and the life quality of human. In view of a series of problems, such as the drug resistance, side effects and poor prognosis, accelerating the development of new anticancer drugs is imminent. Recently, since the discovery of cisplatin with anti-tumor activity, the hot of biochemical research has been gradually focused on the chemistry coordination modes, and the relationship of different coordination modes and biological activity. Therefore, a large number of transition metal complexes were synthesized, and their anti-tumor activity were also screened. Pyrazolone Schiff base and its metal complexes were simply synthesized and stable. It is convenient to have the structural transformation. And because of the unique physical activities, such as anti-inflammatory, anti-microbial, anti-cancer, anti-oxidants, anti-virus, cytotoxicity and free radical scavenging, which have aroused great interest to carry out a widespread, systematic and in-depth theoretical and applied research in new drug research. In this study, the heterocyclic β-diketone double Schiff bases was chosen as ligands to synthesize a kind of novel heterocyclic acyl pyrazolone condensing aromatic amines Schiff base, i.e.(Z)-4-(furan-2-yl(p-tolylamino)methylene)-3-methyl-1-p-tolyl-1H-pyrazol-5(4H)-one. And using MS, 1H-NMR, and other analysis methods to confirm the structure of their bonds, and to investigate its anti-tumor activities in vitro. The main contents were summarized as follows:Objective:To synthesize a kind of novel heterocyclic acyl pyrazolone condensing aromatic amines Schiff base, i.e.(Z)-4-(furan-2-yl(p-tolylamino)methylene)-3-methyl-1-p-tolyl-1H-pyrazol-5(4H)-one. And using the modern means of spectroscopy to characterize its crystal structure. At the same time, the novel heterocyclic acyl pyrazolone condensing aromatic amines Schiff base was performed complex with the transition metal of copper to obtain a new metal complexes, the anti-tumor activity in vitro of which was tested.Methods:1. According to the synthesis principle of Schiff base, firstly, methyl phenylhydrazine hydrochloride was synthesized using toluidine as raw material. Secondly, the methylphenyl hydrazine hydrochloride and Na OH solution(10%) were used to prepare the methyl phenylhydrazine. Next, the methylphenyl hydrazine hydrochloride, ethyl acetoacetate and furoyl chloride were used to synthesize and prepare 1-p-tolyl-3-methyl-4(2-furoyl)-5-pyrazolone, which was used to form a Schiff base ligand with toluidine finally. Then the transition metal ions Cu2+ was selected to react to form a Schiff base complexes. Using the modern testing methods(including MS, 1H-NMR, 13C-NMR, X-ray, elemental analysis and thermogravimetric analysis) to investigate the structure, composition, properties and the bond of the above synthesized Schiff base ligand.2. Using CCK-8 method, the anti-tumor activity of(Z)-4-(furan-2-yl(p-tolylamino)methylene)-3-methyl-1-p-tolyl-1H-pyrazol-5(4H)-one copper complex was screened. We studied its effect on the growth and proliferation, cell apoptosis and cycle of osteosarcoma cells(MG-63). DNA ladder was showed on DNA agarose electrophoresis. The intracellular reactive oxygen species(ROS) content was determined by flow cytometry.Results:1. We synthesized a kind of novel heterocyclic acyl pyrazolone condensing aromatic amines Schiff base, i.e.(Z)-4-(furan-2-yl(p-tolylamino) methylene)-3-methyl-1-p-tolyl-1H-pyrazol-5(4H)-one, the structure of which is not found at home and abroad. Yellow block single crystals suitable for an structural analysis by MS, 1H-NMR, 13C-NMR, X-ray, elemental analysis and thermogravimetric analysis were obtained. And the final yield was 75.6%. At the same time, the transition metal ions Cu2+ was selected to react to form a Schiff base complexes with this Schiff base ligand.2. The preliminary anti-tumor activity in vitro test results indicated that the new Schiff base copper coordination compound could inhibit MG-63 osteosarcoma cells growth when compared with negative control in a doseand time-dependent. CCK-8 assay indicated that when the concentration was 70 μmol/L, the inhibition rate reached 79.41%. The apoptosis rate were determined by flow cytometry. The results indicated that apoptosis rate were(18.2 ± 1.02)%,(27.1 ± 1.15)%,(77.1 ± 1.09)% at a concentration of the new Schiff base copper coordination compound of 10, 30, 70 μmol/L, respectively. The results have significant statistically differences(P<0.05) compared with the control group. The novel Schiff base copper coordination compound could arrest MG-63 osteosarcoma cells at G1/G0 as indicated by flow cytometric analysis. We could see the DNA ladder on agarose gel electrophoresis when detecting cell apoptosis. In addition, the novel Schiff base copper complexes could induce the levels of intracellular ROS increased, confirmed that drugs candestroythe defense system of tumor cell and induce oxidative damage to cells.Conclusion:The synthesized novel heterocyclic acyl pyrazolone condensing aromatic amines Schiff base has been nurtured single crystal, the results of structure characterization shown that it was a kind of new compound. At the same time, the preliminary anti-tumor activity in vitro test of the transition metal copper coordination complexes results shown that the new Schiff base copper coordination compound could inhibit MG-63 osteosarcoma cells growth when compared with negative control in a dose- and time-dependent. The results of this study will lay a good foundation for the further development of the new low toxicity and highly active anti-tumor chemotherapy drugs. At the same time, it will provide a better and more effective choice of chemotherapy drug for the treatment of advanced cancer associated with bone metastases and other organs metastasis. |
PDF Full Text Request