Design, Synthesis And Screening Of CCR4 Antagonists Based On A Novel Cytokine CKLF1

PKU human disease genomics center has identified a novel cytokine CKFL1 by SSH technology in PHA-stimulated U937 cell.It is a member of chemokine-like factor super family.This genefamily is a new human gene discovered firstly by our nation internatio -nally.CKLF1 could introduce phathology change such as the situation during the last period of asthma.The C-terminal of CKLF1 could interact with the same receptor as CKLF1,and shows antagonist activity.It would be of theoretical significance and potential value that research on structure and function of CKLF1.This thesis,in which computer aided drug design method was used,is based on those pharmacological results.3D structure of CCR4 was obtained by homology modeling; conservative residues were used to make sure the multiple sequence alignment accuracy; density map and hydrophobicity plot were used to adjust the orientations of transmembrane regions.3D structures in nature condition of C 19 were got by folding recognition and ab initio prediction.After that,the active sites of CCR4 were predicted by correlation muta -tion analysis.The solvent assess surface of C19 had been analyzed;and the secondary structure stabilization had been studied,which were both used to give the clue of C 19 key residues.Restricted by the electric surface complementary,C19 was docked to CCR4.Key residues of C19,which were frequently combined to CCR4,were found.Those residues constituted a triple peptide.In order to mimic this triple peptide,thiourea and 2-aminothi -zole cores were designed.68 new compounds have been synthesized,all of which were confirmed by MS and ~1H-NMR.Thiourea derivatives were gained through the addition of isothiocyanate and primary amino.2-aminothizole derivatives were prepared by combinating secondary aminos and carbonyl chlorides to 2-amino-4-Chloromethyl-thizole.The routes for prepar -ing those target molecules were optimized at the mean time.Their bioactivities were screened by both of boyden small cell method and capillary electrophoresis method.They showed different potential activity against CCR4.Especially Compound 9,12,23,27 and 38# can interrupt the interaction of all three nature ligands to CCR4. Quantity and structure relationships were analyzed.It is showed that the thiourea structure molecules are better;the best linker length is 3-carbon length;heteroarmatic part is better than aromatic attachment as a side chain.All of these messages could be guidance for further research of CCR4 antagonists.Above all,CADD technology supplies us convinent and reliable strategies from protein sequence to target molecules,and they could make drug research reasonable in dark.