Screening For Lysyl-tRNA Synthetase Inhibitors

Aminoacyl-tRNA synthetase(aaRS)is an important enzyme in organisms.It mainly catalyzes the combination of amino acids and tRNA to generate aminoacyl-tRNA,which is the key enzyme for protein synthesis.Lysyl-tRNA synthetases(LysRS)is one of 20 Aminoacyl-tRNA synthetases in human body.Its main function is to catalyze lysine to combine with its corresponding tRNA to generate lysyl-tRNA and participate in the synthesis of protein peptide chain.Current research shows that parasite LysRS inhibitors such as plasmodium and filariasis can be used as antiparasitic drugs.In addition,the high expression of LysRS in some cancer cells also indicates that lysrs can be used as a target for anticancer drugs.Cladosporin is an inhibitor of LysRS.Studies have found that Cladosporin has antibacterial,insecticidal and anti-inflammatory activities,as well as inhibiting tumor growth.However,more in-depth studies have shown that mycosporin has many constraints such as drug-forming properties.Therefore,this paper aims to screen LysRS inhibitors with better inhibitory effect and drug-forming properties.In this paper,the combination of computer simulation molecules and LysRS was used,and Cladosporin was used as a ligand template to screen 105 small molecular compounds with good combination with LysRS.These compounds were purchased for experimental screening.Firstly,the inhibitory screening of enzyme activity is carried out on the small molecule compounds virtually screened out:the active human LysRS protein is expressed and purified through Escherichia coli;Establish malachite green phosphorus determination method for indirect determination of LysRS activity;After the determination method of enzyme activity is established,the screened small molecule compounds are tested for inhibiting enzyme activity.According to the inhibition rate of small molecule compound on human LysRS enzyme activity,a small molecule compound with good inhibition effect(specsid:an-465/41851406)was finally determined,and its inhibition efficiency on LysRS protein reached 60%at 1 mM concentration.After the target compound is screened for inhibition of enzyme activity,the compound is subjected to cell and in vivo anti-tumor experiments to evaluate the feasibility of being used as a drug lead compound.The results showed that when the small molecule compound AN-465/41851406 was co-cultured with B16 melanoma cells for 48 hours,the cells showed a state of shrinkage and aggregation.When the concentration was 2 mM,the inhibition rate to B16 melanoma cells was 31%.Animal experiments show that the small molecule compound AN-465/41851406 has a certain inhibitory effect on melanoma growth in B16-bearing mice,and the tumor growth rate recovers after drug withdrawal.Experiments show that the small molecule compound AN-465/41851406 can enter cell tissues and has a certain tumor growth inhibition effect,and the compound has the possibility of serving as a drug lead compound.