Study On Chirality Synthesis Of 3-quaternary Carbon-β’-nitroguanidine Synthesis And Activity Evaluation

The p53 gene is the tumor suppressor gene most relevant to human tumors,and the MDM2 protein,an expression product of the oncogene DMD2,binds to the p53protein.In theory,blocking the action of MDM2 on p53 can restore the normal function of p53.The crystal structure of P53-MDM2 complex shows that the purine ring of the tryptophan side chain is the most important binding element of p53 binding MDM2 protein,and screening by computer-assisted drug.The most consistent compound fragment of the anthracycline,cerium oxide,was simulated.Based on the methodology of new drug synthesis and the principle of drug splicing of active skeleton transition,a series of 3-quaternary carbon-’-nitroguanidine compounds with chromone skeleton were efficiently constructed by Michael addition reaction.The synthesized compounds were evaluated in vitro for antitumor activity and preliminary mechanism research.It is expected to find some high-efficiency,low-toxic lead compound skeletons,which provide a certain material basis for the evaluation of antitumor activity in vivo.This thesis consists of three parts:The first part reviews the research progress of3-quaternary carbon-β’-nitroguanidine compounds and anti-tumor drug screening technology;the second part reports 3-quaternary carbon-β’-nitrate The Michael addition reaction of ruthenium oxides and its subsequent expansion studies;the third part reports the in vitro anti-tumor activity and preliminary mechanism of3-quaternary carbon-β’-nitroguanidine compounds.Thefirstpartreviewstheresearchprogressof3-quaternary carbon-β’-nitroguanidine compounds and anti-tumor drug screening technology.At present,anticancer therapeutic drugs generally have toxic side effects,and some tumor cells are also resistant to drugs,and a combination of drugs is needed to have a better effect.Therefore,it is imperative to find new anti-tumor drugs with small toxic side effects,and many compounds with natural product skeleton have better anti-tumor activity,among which cerium oxide compounds are the focus of research.This project designed to splicing several parts of the skeleton into a compound,and structurally modified and modified to synthesize a series of 3-quaternary carbon-β’-nitroguanidine skeleton derivatives,in order to find new drug candidates with better activity.Compounds,at the same time comprehensively expounded the current partial anti-tumor drug screening technology,laying the foundation for follow-up work.The second part reports the results of the Michael addition reaction of3-quaternary carbon-β’-nitroguanidine compounds.In this study,a Michael addition reaction of a ruthenium oxide compound with nitrostyrene was carried out to obtain differently substituted 3-quaternary carbon-β’-nitroguanides.In this study,a 3-quaternary carbon-β’-nitroguanidine compound skeleton was constructed using two components.In particular,the product has high diastereoselectivity and high ee value.Through the Sandmeyer iso-nitrosoguanidine aniline Isatin synthesis method,the initial substrate ruthenium was first synthesized,and then a series of reactions such as Grignard,reduction and Michael addition reaction were carried out to obtain different substituted 3-quaternary carbon-β’-Nitrophosphorus oxide compound.In addition,the reaction time,catalyst,solvent,etc.were screened to obtain an optimal reaction route.In the chiral catalyst1-[3,5-bis（trifluoromethyl）phenyl]-3-[（1S,2S）-（+）-2-（dimethylamino）cyclohexane] thiourea as the catalyst and DCM as the reaction solvent,the reaction at 45°C can obtain a medium to high yield（up to 90%）with good diastereoselectivity（up to 20:1）.A total of 24 target compounds were obtained with a high ee value（up to 98%）.The reaction route is environmentally friendly,economical and efficient,and also provides a method for constructing a 3-quaternary carbon-β’-nitroguanidine compound containing a ketone skeleton.Subsequent work replaced nitrostyrene with N-acetyl compounds,synthesized 11 compounds,and explored the universality of the reaction in order to obtain better anti-tumor drugs.The third part of the work is to evaluate the in vitro antitumor activity of the synthesized compounds.Using the MTT method,the anticancer activity of the synthesized 3-quaternary carbon-β’-nitroguanidine compound was studied in vitro using the anticancer drug cisplatin,which has a broad spectrum of anticancer activity,as a positive control.Inhibition of human chronic myeloid leukemia cell lines.The results showed that the nine compounds 8a,8c,8e,8j,8o,8q,8r,8w and 8x had good inhibitory activity against human chronic myeloid leukemia cell line K562,and the three compounds 8a,8c,8r were non-small to humans.Cell lung cancer cell A549 has good inhibitory activity,and all has good inhibitory activity against human prostate cancer cell line PC-3,and can reach the same order of magnitude as the positive control drug cisplatin(cisplatin as a positive control,its IC₅₀0 concentration is K562:17.58μM;A549:13.84μM;PC-3:21.29μM),showing potential research value.Therefore,the 3-quaternary carbon-β’-nitroguanidine compound can be used as a lead compound for antitumor active drug research.Finally,K562 cells were treated with different concentrations of compound 8a for 24 h.The expression of anti-apoptotic factor Bcl-2 and proapoptotic factor Bax was detected by Western blot.The results showed that the relative expression levels of Bax and Bcl-2 were The K562 cells treated with 8a did not change significantly after 24h,indicating that the apoptosis of K562 cells induced by compound 8a may not depend on the expression of Bax and Bcl-2.