Virtual Screening Of Dual-target Inhibitors Of FTase And Raf-1

Computer-Aided Drug Design(CADD)is an important method for modern drug development.It requires the least compound design and prior knowledge,but it can screen out a variety of candidate compounds,and greatly save experimental funds.In this study,we aim to explore a highly selective and highly active dual-target tumor inhibitor through CADD.With the development of molecular biology and other related displines,the research of anti-tumor drugs has been transformed to new anti-tumor drugs targeting at the multi-link mechanism of tumor development.Some tumors can develop resistance to a single protein or kinase inhibitor.In order to maximize its efficacy,multi-target drug use has become a mature treatment method.Compared with single-target drug therapy,dual-target drug therapy has higher efficacy and can reduce the treatment dose and side effects.Ras protein plays an important role in the activity of several key signaling pathways that regulate normal cell proliferation.Studies have found that about 30% of tumorigenesis is mediated by mutations in Ras gene,and a single point mutation of Ras gene is enough to trigger malignant transformation of tumors.Therefore,therapies targeting Ras proteins and the signaling pathways they control will be valuable in the treatment of tumors that activated Ras mutations.Ras proteins need to undergo post-translational modification to function,and farnesyltransferase(FTase)is the first key enzyme in this process.In addition,Raf-1 kinase is a key kinase downstream of the Ras signaling pathway and can regulate the cells without being dependent on the Ras signaling pathway.Therefore,inhibition of the downstream RAF-1 kinase while inhibiting FTASE can not only greatly enhance the anti-tumor activity of a simple FTase inhibitor,but also expand the type of anti-tumor.Combined with the concept of multi-target drug design,multi-target drugs that can act on the above two targets were designed with FTase in the RAS signaling pathway as the central target and its downstream Raf-1 kinase as the auxiliary target.We used CADD technology to search for dual-target inhibitors for the treatment of tumors.For this purpose,we made three parts.In the first part,we screened the single target inhibitors of FTase.The ligand-based computational method was used to determine the molecular chemisty needed to inhibit FTase.The pharmacophore model was generated from the training set database of known inhibition activities using the 3D Quantitative Struct-Activity Relationship(3D QSAR)pharmacophore generation module provided by Discovery Studio 2020(DS2020).The model was verified by four different methods,and the best model Hypo1 was applied to screen three databases of Minimay Bridge,Druglike and Traditional Chinese Medicine.The molecular docking module was used to conduct molecular docking research on the compounds with high inhibitory activity retrieved,and 10 best drug candidates were obtained by comprehensive analysis of various indicators.Combined with in vitro experiments,the best drug candidate molecule CDI909983 was obtained.In the second part,we screened the single target inhibitors of Raf-1.A training set database of 18 compounds was used to generate pharmacophore groups using the Hypogen algorithm based on activity prediction ability.The prediction ability was further verified by test set database,Fischer stochastic method,cost value detection and other methods.The pharmacophore Hypo1 with the best predictive ability contains the chemical characteristics required to effectively inhibit Raf-1 kinase,and the best model was selected to screen the database.The 135 drug molecules obtained by Lipinski’s five rules and minimum activity restriction were linked to the active center of Raf-1 kinase,and VIT169751 was considered as the best drug candidate.In the third part,FTase and Raf-1 dual target inhibitors were screened.We have obtained the optimal pharmacophore model of two single targets through verification.Based on the principle of stratified screening and according to the strategy of dual-target inhibitor screening,we have carried out screening including pharmacophore modeling,virtual screening,molecular docking,Veber rule,ADMET and so on.Finally,three potential dual-target inhibitors,UKR24453,CDI909983 and VIT169751,were identified that act on both FTase and RAF-1 kinases.Considering the complexity of tumor causing elements,CADD can help develop novel drugs with multiple targets,which will be an effective way to develop tumor drugs.Through three parts of FTase single target inhibitors,Raf-1 single target inhibitors and FTase and Raf-1 double target inhibitors,10 compounds ranked best were screened by CADD technique,and all the calculated compounds showed excellent inhibitory activity.CDI909983 is the best single target inhibitor for FTASE,VIT169751 is the best single target inhibitor for Raf-1.UKR24453,CDI909983 and VIT169751 are the best double target inhibitors for FTase and Raf-1.