Studies On Synthesis,Structure And Antitumor Activity Of Four Kinds Of The Novel Anthracene Hydrazone Derivatives And Their Copper,Gold Complexes

Based on the structure of bisantrene,four novel anthracycline drugs and coordinated with Cu（Ⅱ）and Au（Ⅲ）Metal ions,five complexes were synthesized and fully characterized by ESI-MS,elemental analysis,NMR,and single crystal X-ray diffraction analysis.At the cellular level,the antitumor activities of ligands and their metal complexes were preliminary studied by MTT assay.Flow cytometry was used to examine cell cycle distribution and cell apoptosis induction and acute toxicity in mice.At the molecular level,the molecular mechanism of the compounds and DNA as well as topoisomerase Ⅰ were studied by competitive binding fluorescence emission spectroscopy and agarose gel electrophoresis experiments.Above work provides a fundamental basis and experience for the research of novel anthracycline antitumor drugs.The contents as follows:The four novel anthracycline ligands are:9,10-APH,9,10-ADMPH,9,10-AATH,9,10-AAIH.Their corresponding metal complexes are:9,10-APH-Cu,9,10-ADMPH-Cu,9,10-AATH-Cu,9,10-AAIH-Cu and 9,10-AAIH-Au.The structures of all the compounds were determined by ESI-MS,Elemental analysis,NMR,and X-ray single crystal diffraction analysis.The stability of the nine compounds were examined by UV-Vis absorption spectra.The figures demonstrated that the absorption peaks without blue shift or red shift,as well as no new absorption peaks appeared in 0 to 48 hours.The results showed that all compounds can stably exist in the solution without precipitation.Firstly,at the molecular level,all compounds were primarily studied for the typical targets of anthracycline antitumor drugs of DNA and topoisomerase by competitive binding fluorescence emission spectroscopy and agarose gel electrophoresis experiments.The results showed that the compounds exhibited a certain degree of insertion with DNA,which was consistent with the molecular mechanism of classical anthracycline antitumor drugs.From the agarose gel electrophoresis experiments of compound-DNA-Topo Ⅰ ternary systems with different mixing sequences,it was found that the compounds described weak inhibition of Topo Ⅰ activity and only exhibited a certain activity inhibition at higher concentrations.Therefore,speculated that Topo I should not be a key target of the metal complexes.Secondly,at the cellular level,the compounds were tested on eight cells（MGC-803,T-24,Hep-G2,BEL-7402,A549,SK-OV-3,He La229,WI-38 and HL-7702）by MTT in vitro proliferation inhibitory activity.Through its corresponding inhibition rate and IC₅₀ value that the five metal complexes exhibited high proliferation inhibitory effect on most tumor cell lines and were higher than the corresponding ligands and the clinical anti-cancer drug of cisplatin,which showed a good prospect,but the activity was still significantly lower than Bisantrene,while it was also more toxic to the normal cells WI-38 and HL-7702.Flow cytometry was used to study the cell cycle arrest and apoptosis of the complexes 9,10-APH-Cu and 9,10-ADMPH-Cu.The results showed that the effect of the two complexes on the four tumor cells for 48 hours were different from the blank group.The He La229 and MGC-803 cells were arrested in the S phase.The SK-OV-3 cell was arrested in G2 phase,and the BEL-7402 cell was arrested in G1 phase,indicated that the complexes had different mechanisms of cell cycle arrest in different types of cells.The results of apoptosis showed that the morphological detection of complex 9,10-APH-Cu can induce cell apoptosis.He La229 and BEL-7402 cells were induced in late apoptosis by9,10-ADMPH-Cu,MGC-803 and SK-OV-3 cells were induced in early apoptosis,They can cause apoptosis in different tumor cells and had different induction apoptosis results.Complexes also had different mechanisms of apoptosis in different tumor cells,may be associated with different cycle arrest mechanisms.Finally,the 9,10-ADMPH-Cu was further tested for acute toxicity in mice.The results showed that the LD₅₀ value was 108.39 mg/kg.The 95%confidence limit was 78.47 to 149.62mg/kg.The complexes of 9,10-ADMPH-Cu and 9,10-APH-Cu were administered at 25 mg/kg.The mice did not show obvious poisoning.It was initially demonstrated that the administration was safe at this dose,the median lethal dose data had to be further studied.In summary,a variety of novel anthracycline drugs were synthesized and characterized.The preliminary study on the anti-tumor activity,mechanism and acute toxicity were conducted at the cell level,molecular level and animal level.Two kinds of anti-tumor compounds with high activity and clear mechanism were found,and further research was expected.This study provided scientific data and research basis for a more in-depth and comprehensive study of the mechanism of novel antitumor compounds based on quinone structures.