Study On The Synthesis And Anticancer Activity Of A Two-target EGFR/PI3K Inhibitor Based On Chrysin Skeleton

Natural products have always been one of the important sources of leading compounds for tumor therapy.Chrysin,also known as 5,7-dihydroxyflavone,is a natural product derived from the seeds and stem bark of Oroxylum indicum(L.)Vent.,a plant of the bignonia family.It has good physiological activities and its anti-tumor activity especially eye-catching.The occurrence and development of tumors are regulated by a variety of proteases in a variety of signal transduction pathways during cell growth.EGFR(epidermal growth factor receptor)is an important type of RTKs(receptor tyrosine kinases),which is involved in activating a variety of signal transduction in cells.Its high expression or abnormal expression will cause Stimulate its downstream pathway PI3K(phosphatidylinositol3-kinase,phosphatidylinositol-3 kinase)/Akt(protein kinase B,protein kinase B,PKB)/m TOR(mammalian target of rapamycin)abnormal expression,resulting in the occurrence and development of tumors.As a natural EGFR inhibitor and PI3K/Akt signaling pathway blocker,Chrysin has been favored by researchers for its anti-tumor effect.However,its disadvantages such as poor solubility and low bioavailability affect its anti-tumor activity.By modifying the structure of chrysin,changing its lipid-water partition coefficient and improving its bioavailability,its anti-cancer activity can be fully exerted,and it has been developed into an anti-cancer drug with high efficiency,low toxicity and good targeting.This idea is worthy of in-depth exploration by researchers.In this study,on the basis of literature research,an amino acid was introduced at the 7-position of chrysin through a hydrocarbyl chain to obtain chrysin-amino acid derivatives.The biological activity of the chrysin-amino acid derivatives was studied,and compounds with better activity were screened out.Meanwhile,Sybyl2.0 software was used to study the structure-activity relationship of the compounds,and 3D-QSAR model was established by Co MFA and Co MSIA methods to predict the activity of these compounds.The structure-activity relationship diagram of the compound against MGC-803 cells was obtained,which provided a basis for the development of lead compounds with potential activity.The main research results are summarized as follows:Chapter 1:This chapter briefly introduces that the occurrence and development of tumors are related to abnormal signal transduction pathways during cell growth.Signal transduction pathways are regulated by a variety of proteases(such as EGFR,PI3K,etc.),and their abnormal expression will lead to abnormal signal transduction pathways.At the same time,this chapter briefly reviews the research status of anti-cancer drugs targeting these proteases,and expounds the research basis and content of this thesis.Chapter 2:We designed the synthetic route of chrysin-amino acid derivatives and formulated the research plan of its structure-activity relationship.According to the principle of drug combination,the natural product chrysin was used as the parent,36 chrysin amino acid derivatives were successfully obtained with the drug synthesis reactions such as Williamson ether synthesis,hydrolysis,and condensation were used to introduce low toxicity and better biocompatibility amino acid into chrysin7-OH,and their purity was determined by measuring the melting point of the compound,TLC method,etc.,and the structure of the compound was confirmed by ¹H-NMR,¹³C-NMR,HR-MS.Chapter 3:Taking MGC-803(human gastric cancer cells),Hela(human cervical cancer cells),MCF-7(human breast cancer cells),A549(non-small cell lung cancer cells),Hep G-2(human liver cancer cells)as the research objects,the anti-cancer properties of the compounds were screened by MTT method and CTG method in vitro,and their toxicity to normal cells HUVECs(Human Umbilical Vein Endothelial Cells)was investigated at the same time.FCM was used to detect the compound's influence on cell apoptosis and cell cycle,Western Blot method and ELISA method were used to study the mechanism of compound inducing cancer cell apoptosis.Surflex-DOCK molecular docking technology is used to study the interaction mode between compounds and target proteins.Chapter 4:based on the above research,combined with the previous research results of this research group,the activity data of 125 compounds inhibiting different cancer cells(MGC-803,Hep G-2,MCF-7,A549,Hela,)were analyzed.The Co MFA method and Co MSIA method are used for calculation,and multiple Co MFA method and Co MSIA models are established by single field and combined field respectively.The PLS analysis method is used to evaluate the model fitting ability and predictive ability.The results of the study found that the compound has an ideal 3D-QSAR model for the activity data of MGC-803,and the statistical indicators of the model(Fisher test value F is greater than 100,standard deviation s approaches 0,correlation coefficient r approaches 1,cross-validation The correlation coefficient q2 is greater than 0.5),which indicates that the model has better fitting ability and good predictive ability.The 3D-QSAR structure-activity relationship of the compound's inhibitory activity on MGC-803 was preliminarily clarified through the three-dimensional equipotential diagram analysis of the model's three-dimensional field,electrostatic field,hydrophobic field,and hydrogen bond(donor)body fields.The results showed that a better 3D-QSAR model was obtained for mg C-803 activity data.Through the analysis of Co MFA model,Co MSIA model and activity prediction results,it was cleared that the IC50 value of the derivatives decreased with the extension of alkanes chain and the increase of amino acid volume,but the IC50 value increased with the increase of upper branch chain of alkanes chain.At the same time,the presence of alkane chain and hydrophobic substituents around amino acids could improve the antitumor activity of the compound,and the structure-activity relationship diagram of the compound inhibiting the activity of MGC-803 cells was obtained.It was expected to develop a lead compound with anti-MGC-803 cell activity through reasonable modification of the active skeleton structure of albumin.