Synthesis Of Aloe-Emodin ?-Aminophosphonate Derivatives And Their Interaction With DNA

ObjectivesBased on the principle of combination of pharmacophores,a series of?-Aminophosphonate Derivatives were designed and synthesized on the basis of aloe-emodin structure.The structure-activity relationship of its anti-tumor activity and its binding mode with DNA were discussed.The purpose of this study is to provide a theoretical basis for the discovery of novel antitumor drugs targeting DNA.MethodsA series of novel aloe-emodin?-aminophosphate derivatives were synthesized by oxidation,condensation and imine addition.The structures were characterized by HRMS,IR,¹H NMR and ¹³C NMR,and the inhibitory effects of derivatives on the growth of lung cancer?A549?,triple negative breast cancer?MDA-MB-231?and hepatocellular carcinoma?HepG2?cells were determined by MTT assay.Confocal microscopy was used to observe the distribution of aloe-emodin,Derivative A and B in MDA-MB-231 cells.The interaction between derivatives A,B and calf thymus DNA?Ct-DNA?was studied by UV spectrum,fluorescence spectrum,DNA thermal denaturation method and viscosity method.Results?1?Thirteen?4,5-dihydroxy-9,10-anthraquinones-2-yl??substituted aniline?methyl phosphate diethyl ester derivatives?A-M?were successfully synthesized.The structures were confirmed by HRMS,IR,¹H NMR and ¹³C NMR.?2?After 13 derivatives treatment of lung cancer cell line A549,breast cancer MDA-MB-231 and liver cancer cell line HepG2,the results of MTT showed that the other derivatives had different degrees of inhibitory effect on the three kinds of tumor cells except derivatives F,H,J,K,The inhibitory effect on lung cancer A549 and HepG2 cells was better than that of breast cancer MDA-MB-231 cells.In particular,derivatives A had significant inhibitory effect on the three cell lines,its IC₅₀0 was?10.89±1.27??mol/L,?14.50±3.87??mol/L and?6.47±2.51??mol/L,respectively.The inhibition was stronger than that of aloe emodin.The inhibitory effect of derivative B on A549 was stronger than that on the other two cells,and the IC₅₀0 of derivative B was?7.54±1.13??mol/L.?3?The distribution of derivatives A and B in cells was observed by confocal microscope.The results showed that derivatives A and B could be uniformly distributed in the cytoplasm,some of them entered the nucleus and a few gathered around the nuclear membrane after treatment withMDA-MB-231cells 30min.Unlike the derivatives A and B,aloe-emodin is distributed in the cytoplasm and hardly enter the nucleus.?4?The results of UV spectrum indicated that the derivative A can cause red shift and the increase absorption of DNA UV spectrum,and compound B only has hyperchromic effect.The hyperchromic rates of derivative A and B were31.0%and 11.9%,binding constants K₀ were 0.87×10⁴ mol/L and 1.44×10⁴mol/L,respectively.When the derivative A and B was added,the melting temperature of Ct-DAN was increased 11?and 7?,respectively.The results of fluorescence analysis showed that the fluorescence intensity of EB-DNA system decreased by more than 50%with the addition of derivatives A and B.At the same time,both derivatives A and B can increase the viscosity of DNA.Conclusions?1??4,5-dihydroxy-9,10-anthraquinones-2-yl??substituted aniline?methyl phosphate diethyl ester derivatives were synthesized successfully.?2?Aloe-emodin?-aminophosphate derivatives have different degrees of inhibitory activity on three kinds of tumor cells,among which derivatives A and B have the strongest inhibitory effect,are superior to aloe-emodin.?3?Derivatives A and B may interact with DNA in intercalation mode.