title: Dual targets guided screening and isolation of Kukoamine B as a novel natural anti-sepsis agent from traditional Chinese herb Cortex lycii.
abstract: Treating sepsis remains challenging at present. Bacterial lipopolysaccharide (LPS) and bacterial DNA/CpG DNA are important pathogenic molecules and drug targets for sepsis. It is thus a promising strategy to treat sepsis by discovering agents that neutralize LPS and CpG DNA simultaneously. In this study, we present evidences of the biosensor based screening and isolation of active anti-sepsis fractions and monomers from traditional Chinese herbs using dual targets (LPS and CpG DNA) guided drug discovery strategy. Firstly, LPS or CpG DNA was immobilized on surfaces of cuvettes in the biosensor to establish a screening platform. Then, Cortex lycii with both highest affinities was selected out from one hundred and fourteen traditional Chinese herbs. In subsequent experiments, chromatography was utilized and coupled with the biosensor to purify fractions with a higher affinity for LPS and CpG DNA. In line with affinity assay, these fractions were shown to neutralize LPS and CpG DNA and inhibit their activity in vitro and in vivo. Lastly, the contributing monomer Kukoamine B (KB) was purified. KB neutralized LPS and CpG DNA in vitro. It inhibited TLR4, TLR9 and MyD88 mRNA expressions up-regulated by LPS and CpG DNA, and also attenuated the LPS and CpG DNA elicited nuclear translocation of NF-κB p65 protein in RAW264.7 cells. It also protected mice from lethal challenge of heat-killed E. coli, a mixture of LPS and CpG DNA. In conclusion, we presented a dual target guided discovery of a novel anti-sepsis agent KB from traditional Chinese herbs via combination of biosensor technology and chromatography methods.
authors.affiliation: Medical Research Center, Southwestern Hospital, The Third Military Medical University, Chongqing 400038, PR China.
authors.forename: Xin
authors.initials: X
authors.lastname: Liu
authors.forename: Xinchuan
authors.initials: X
authors.lastname: Zheng
authors.forename: Yupeng
authors.initials: Y
authors.lastname: Long
authors.forename: Hongwei
authors.initials: H
authors.lastname: Cao
authors.forename: Ning
authors.initials: N
authors.lastname: Wang
authors.forename: Yongling
authors.initials: Y
authors.lastname: Lu
authors.forename: Kecen
authors.initials: K
authors.lastname: Zhao
authors.forename: Hong
authors.initials: H
authors.lastname: Zhou
authors.forename: Jiang
authors.initials: J
authors.lastname: Zheng
mesh_terms: D000818:Animals; D015374:Biosensing Techniques; D002109:Caffeic Acids / Q000302:isolation & purification / Q000494:pharmacology; D002460:Cell Line; D018899:CpG Islands / Q000187:drug effects / Q000235:genetics; D004269:DNA, Bacterial / Q000737:chemistry* / Q000187:drug effects; D055808:Drug Discovery / Q000379:methods; D004365:Drugs, Chinese Herbal / Q000302:isolation & purification / Q000494:pharmacology; D008070:Lipopolysaccharides / Q000737:chemistry*; D008264:Macrophages / Q000187:drug effects / Q000276:immunology; D051379:Mice; D018805:Sepsis / Q000188:drug therapy / Q000276:immunology; D013096:Spermine / Q000031:analogs & derivatives* / Q000302:isolation & purification / Q000494:pharmacology
chemical_list: D002109:Caffeic Acids; D004269:DNA, Bacterial; D004365:Drugs, Chinese Herbal; D008070:Lipopolysaccharides; C557784:kukoamine B; C440499:lipopolysaccharide, Escherichia coli O111 B4; D013096:Spermine