Synthesis Of 5-fluorouracil Bond With Dithiocarbamate Derivatives And Research On Its Antitumor Activites In Vitro

5-fluorouracil is a commonly used first-line anti-tumor drug of pyrimidine in clinical.It has a wide anticancer spectrum,but it has the disadvantages of short half-life and low selectivity.Therefore,people are trying to find a prodrug of 5-fluorouracil with better selectivity and lower side effects.Dithiocarbamate derivatives have strong ability of chelating metal ions,the anti-tumor mechanisms of DSF have been elucidated,the family of the same in the body as copper ions to form stable complex,this complex can be strongly combined with NPL4 protein,making a lot of defect proteins in the tumor cells cannot be remove,eventually induce the apoptosis of tumor cells.In this paper,with the guidance of the principle of drug assembly,5-fluorouracil was bonded with dithiocarbamate derivatives,and a series of in vitro antitumor activity evaluations were conducted,in order to obtain potential antitumor precursor drugs.1.5-fluorouracil reacted with bromoacetic acid to prepare the intermediate5-fluorouracil-1-yl acetic acid,and then acylates 5-fluorouracil-1-acetyl chloride by thionyl chloride.Then respectively with sodium diethyl dithiocarbamate,pyrrole alkyl ammonium dithiocarbamate,ethyl phenyl disulfide generation amino zinc and(N-ethyl)piperazine reaction synthesis of four different split things [2-(5?-fluorouracil)acetic acid-diethyl dithiocarbamate] anhydride(Compound P1 for short,the same as below),[2-(5?-fluorouracil)acetic acid-pyrrole alkyl dithiocarbamate] anhydride(Compound P2 for short,the same as below),[2-(5?-fluorouracil)-ethyl phenyl acetate dithiocarbamate]anhydride(Compound P3 for short,the same as below),[2-(5?-fluorouracil)acetate-(N-ethyl)piperazine dithiocarbamate] anhydride(Compound P4 for short,the same as below),the structure of each product was confirmed by 1H-NMR and13C-NMR.2.The cytotoxicity test was carried out by MTT method,the results showed that the inhibitory effect of the four conjugations on tumor cells was not significantly stronger than that of the positive control group 5-FU,but when the conjugations were used in combination with copper ions,the anti-tumor activity of the four conjugations was significantly stronger than that of the control group 5-FU/Cu(5-FU at equal molality was administered in combination with copper gluconate),and as for B16,HepG2,MCF-7 and U87 MG cells,P3/Cu(P3 at equal molality was administered in combination with copper gluconate)and P4/Cu(P4 at equal molality was administered in combination with copper gluconate)has stronger anti-tumor effects than P1/Cu(P1 at equal molality was administered in combination with copper gluconate)and P2/Cu(P2at equal molality was administered in combination with copper gluconate),and P3/Cu has the strongest anti-tumor effects on A549 and BGC823 cells.3.The colony-formation assays was carried out by crystal violet staining,the results of crystal violet staining showed that the clone formation rate of tumor cells decreased significantly with the increase of the dose concentration,indicating that theconjugations combined with copper ions can inhibit the proliferation of tumor cells.4.Transwell plate was used for cell transfer experiments,the results showed that with the increase of drug concentration,the number of cells transferred to the lower surface of the upper compartment decreased in turn.5.Annexin v-fitc /PI double staining method was used for apoptosis assay,the results showed that P3/Cu and P4/Cu could promote the apoptosis of MCF-7 and U87 MG cells,and the two drugs had a stronger effect on the apoptosis of MCF-7 cells than that of U87 MG cells.6.Cell cycle experiments were carried out by PI staining,the results showed that P3/Cu and P4/Cu could cause G1 phase block of MCF-7 and U87 MG cells,and P3/Cu had a stronger G1 phase block effect on tumor cells than P4/Cu.7.Molecular docking technology was used to study the docking between compound molecules and target proteins.We use molecular docking technology to simulate four split at the rear of the metabolic and copper ions to form complex,the four kinds of complex docking with NPL4 protein,the results showed that P3 split in the metabolism of copper ion complex content and NPL4 protein has the best effect of affinity,followed by P4,P1 and P2 are relatively weak.In summary,the conjugations combined with copper ions,especially P3/Cu,can significantly inhibit tumor cells after use,and can play an anti-tumor role by inhibiting the proliferation activity of tumor cells,inhibiting cell metastasis,causing cell apoptosis and causing cell G1 phase block.Therefore,this kind of congjugations,especially P3,has the value of further research and exploration.