Design Synthesis And Biological Evaluation Of Small Molecule Inhibitors Targeting Two Oxidoreductases

Redox enzymes are a class of enzymes that catalyze redox reactions.It plays an important role in the oxidative production capacity of organisms,detoxification,and the formation of certain physiological substances.It belongs to the first category of enzymes and it exists in a large number in the biological world.Among them,cyclooxygenase(COX-2)and enoyl-acyl carrier protein reductase(Inh A)are important targets in the field of medicinal chemistry in recent years.Cyclooxygenase(COX)is the rate-limiting enzyme in prostaglandin biosynthesis.There are two subtypes,COX-1 and COX-2.COX-1 is a structural enzyme that catalyzes the production of physiological prostaglandins to maintain normal physiological functions,while COX-2 is an inducible enzyme that is induced by stimulants such as pro-inflammatory cytokines and oncogenes,and is undetectable in most normal tissues.Non-steroidal anti-inflammatory drugs are a class of drugs with anti-inflammatory,analgesic,antipyretic and other effects.Its therapeutic effect is accomplished by inhibiting cyclooxygenase,However,long-term treatment with traditional non-steroidal anti-inflammatory drugs will cause side effects such as gastrointestinal damage,bleeding,and liver toxicity due to the inhibition of COX-1.At the same time,COX-2 can be induced and overexpressed in several solid malignanttumors such as breast cancer,prostate cancer and colorectal cancer.Many reports indicate that the overexpression of COX-2 is related to the occurrence of carcinogens,tumor proliferation,metastasis and invasion.Therefore,the development of COX-2selective inhibitors has become the focus of current research.Inh A is an enoyl-acyl carrier protein reductase in the type II dissociated fatty acid biosynthesis pathway(FAS-II system)in Mycobacterium tuberculosis,which catalyzes the double bond reduction of nicotinamide adenine dinucleotide that depends on the reduced state This double bond is located at position 2 of the growing fatty acid chain connected to the acyl carrier protein.Inh A is a reliable target for the first-line drug isoniazid in the treatment of tuberculosis.Tuberculosis drugs discovered through common routes such as protein synthesis,cell walls,DNA gyrase and ATP synthesis face problems including poor drug resistance and poor selectivity,and blocking fatty acid biosynthesis through Inh A is an effective method of tuberculosis drug design.This subject targets COX-2 and Inh A respectively.We use computer-aided drug design to design and screen new anti-tumor drugs that can target COX-2 inhibitors and new antibacterial drugs that can target Inh A inhibitors based on the existing active structure.Finally,88 phenylpyrazole derivatives and 23 rhodanine derivatives were synthesized.The structure was confirmed by 1H NMR,13 C NMR,ESI-MS,etc.The target compounds were all reported for the first time.Regarding the selective inhibitor of COX-2,compounds Y1-6,Y2-6,Y4-19 showed the most effective anti-proliferative effect of this series in vitro.The IC50 of Y1-6 were 4.61 ?M,10.81?M,and 2.09 ?M for He La,MCF-7,and MDA-MB-231 cells respectively.The results of enzyme-linked immunoassay in vitro showed that the synthesized compounds had good COX-2 inhibitory ability,and the IC50 ??of compounds Y1-6,Y2-6,Y3-21,and Y4-19 were 0.17 ?M,0.31 ?M,0.43 ?M,0.22 ?M.Subsequently,mechanistic studies showed that compound Y1-6 can induce an increase in the level of reactive oxygen species in tumor cells and induce apoptosis of MDA-MB-231 cells and block the cell cycle in the G2 / M phase.ADMET analysis showed the virtual drugability of Y series,and molecular docking also revealed the possible bindingmodes between compounds Y1-6,Y2-6,Y3-21,Y4-19 and COX-2.These results indicate that Y series compounds have the potential to be selective inhibitors of COX-2,and may be further developed as new anti-tumor drugs.Regarding to the part of Inh A inhibitor,the compound RB23 has an activity against M.tuberculosis enoyl-acyl carrier protein reductase inhibitory activity and M.tuberculosis growth-blocking activity equivalent to the positive controls triclosan and isoniazid.Its toxicity to normal cells is also very low.ADMET analysis shows the virtual drugability of RB series compounds.Docking simulation revealed possible binding modes,where key residues were selected as Ser20,Phe149,Lys165 and Thr196.All in all,this study designed a series of compounds based on the two oxidoreductases COX-2 and Inh A,many of which showed good activity,providing new design ideas for the development of anti-tumor and antibacterial drugs,and it has broad development prospects.