Synthesis Of Tanshinone ⅡA Derivatives And Inhibit The Proliferation Of Lung Cancer Cells By Target The KRAS G-Quadruplex DNA

Cancer,as an important disease affecting human survival,has always been one of the important studies.However,lung cancer,it has high mortality and morbidity among cancers.Surgical treatment is the most important and effective method in early stage lung cancer.Nevertheless,many lung cancer patients diagnosed at an intermediate or even advanced stage,whereas surgery was not a good treatment,and the chemotherapy and radiation therapy became the main treatment.In recent years,with the discovery of many oncogenes in lung cancer and the activation of some oncogenes is considered to be one of the factors affecting the occurrence of lung cancer,drug therapy targeting oncogenes provides a better choice for prolonging the survival of lung cancer patients.However,there is one gene in lung cancer that has long been a nightmare for lung cancer patients,this is the RAS gene.As the earliest driver gene of lung cancer,the therapeutic and prognostic effects of lung cancer patients with RAS gene activated are very poor.RAS gene family includes NRAS,HRAS and KRAS,among which KRAS is the most common oncogene(accounting for about 85% of RAS mutations).According to statistics,about 29.61% of patients with non-small cell lung cancer(NSCLC)carry KRAS gene mutations.In recent decades,natural products were study as potential anti-tumor inhibitors.For example,tanshinone-ⅡA(Tan-ⅡA),one of the active components extracted from Chinese traditional medicine Salvia miltiorrhiza,have been found to play anti-inflammatory,treat coronary heart disease,fight atherosclerosis and inhibit the proliferation of cancer cells.However,Tan-ⅡA,a lipid soluble component,has the disadvantages of poor water solubility and low bioavailability,which affects its development as a potential anti-tumor drug.Therefore,structural modification of Tan-ⅡA to change its chemical properties is one of the research directions to improve the anti-tumor effect.G-quadruplex DNA,a special secondary structure of DNA,is aboundant in the human genome,such as proto-oncogene promoter regions,telomeres or aptamers.The formation of G-quadruplex DNA is affect by the number of nucleotide chains,the polatity of G-quadruplex DNA,the types of rings and cations formed.In addition,the formation of G-quadruplex DNA affects the replication,transcription and translation of DNA.In recent years,it has been demonstrated that G-quadruplex DNA targeting oncogene promoter regions can inhibit tumor proliferation.Currently,the G-quadruplex DNA were used as a potential target for anticancer drugs.The promoter region of proto-oncogene KRAS has been found to form the secondary structure of G-quadruplex DNA.Therefore,the Tan-ⅡA derivative targeting KRAS G-quadruplex DNA to inhibit the expression of KRAS oncogene,which may become one of the effective targeted drugs for the treatment of lung cancer.The research of this paper is as follows:(1)A Tan-ⅡA derivative 1 designed and synthesized by microwave assisted synthesis technique.The cell assay results showed that Tan-ⅡA derivative 1 had better inhibitory against non-small cell lung cancer A549 and SPC-A-1 cells than other cancer cells,and can inhibit the proliferation of A549 and SPC-A-1 cells in vitro and in vivo.The mechanism studies have shown that compound 1 can induce DNA damage in NSCLC cells and induce G2/M phase arrest and apoptosis.In addition,toxicity test data of zebrafish indicated that Tan-ⅡA derivative 1 had weak toxicity in vivo.Therefore,Tan-ⅡA derivative 1 may be a potential inhibitor of non-small cell lung cancer.(2)A Tan-ⅡA derivative 2 designed and synthesized by microwave assisted synthesis technique.The results of MTT assay showed that Tan-ⅡA derivative 2 have better activity against lung cancer cells than Tan-ⅡA derivative 1.The colony formation experiment showed that compound 2 could inhibit the growth of four lung cancer cells in vitro,and it can inhibit the growth of SPC-A-1 cells in zebrafish.The mechanism studies have shown that Tan-ⅡA derivative 2 can induce s-phase arrest and apoptosis of different lung cancer cells,which may be caused by DNA damage of lung cancer cells.In addition,Tan-ⅡA derivative 2 has stronger binding ability to KRAS G-quadruplex DNA than other G-quadruplex DNA and double-stranded DNA,and can reduce the expression of KRAS gene and KRAS protein in different lung cancer cells.Therefore,Tan-ⅡA derivative 2 may inhibit the proliferation of different lung cancer cells by targeting KRAS G-quadruplex DNA.In general,two Tan-ⅡA derivatives 1 and 2 were synthesiz.The derivatives 1was found to be effective in inhibiting proliferation of non-small cell lung cancer cells.The Tan-ⅡA derivative 2 can bind KRAS G-quadruplex well and reduce the expression of KRAS gene and protein in lung cancer.Therefore,Tan-ⅡA derivative 1can be used as a potential inhibitor of non-small cell lung cancer proliferation,and Tan-ⅡA derivative 2 can be used as an anti-lung cancer cell proliferation inhibitor and may be used in the treatment of KRAS mutant lung cancer in the future.