Design,synthesis And Bioactivities Evaluation Of Novel Small Molecule ROR?t Inhibitors

Autoimmune disease?AID?is a chronic inflammatory disease in which the body produces an immune response to its own antigen and then results in causes tissue and organ damage.Recent study disclosed that Th17 cells in a variety of autoimmune diseases also play an important role and widely participate in the occurrence and development of all kinds of a variety of autoimmune diseases.Inflammatory cytokine?interleukin-17,IL-17?secreted by Th17 cells can trigger inflammation or cause the occurrence of autoimmune diseases.Retinoic acid orphan receptor?ROR?t?has been concerned for its ability to influence Th17 cell development,and the absence of ROR?t has been found to cause Th17 cell differentiation associated with the cytokine production to stop.ROR?t inhibitors have been reported to inhibit Th17 cell differentiation and IL-17 production in vitro.Hence,ROR?t small molecule inhibitors have become the research hotspot of many research institutions and famous pharmaceutical companies in the field of autoimmune diseases worldwide.In the thesis,ROR?t is selected as targets and the compounds with better activity reported by Vitae compouny in patent WO2014179564 as lead coupounds.I expore the ROR?t small molecule inhibitors with the new skeleton by the application of basic theory of drug design,combined with computer aided drug design methords.The designer inhibitors have been synthesized and characterized by a series of physical methords.The biological activities have been evaluated and the structure-activity relationship analysis,has been dicussed in details.Some highly active small molecule inhibitors have been obtained.The specific research contents and results are as follows:?1?Based on the three-dimensional structure of the eutectic complex formed by ROR?t-LBD protein and small molecule inhibitor VTP-43742,the binding interaction of VTP-1?published in the Vitae patent?and ROR?t was explored and the results suggested to be basically the same as that of VTP-43742 and ROR?t-LBD.The structural characteristics of VTP-1compound was analyzed,and different modifications were made from the group of the head,the parent nucleus and the amide.Three kinds of compounds were obtained:thiazole-pyrrole,phenylacetylene and phenylarylheterocyclic compounds.?2?Twenty compounds in three categories were synthesized.Compounds 1a,1b and 1c in pyrrole series were synthesized by taking different benzenesulfonyl derivatives as key intermediates and through amidation condensation and reductive amination as key reactions.With p-ethyl sulfonyl benzyl alcohol or amine as the key intermediate,carbamate or ureas were constructed by phenyl chloroformate.Compounds 2a and 2b in phenylacetylene series were contrasted through amide condensation and sonogashira reasction.Compounds 3a,3b,3c,3d were synthesized by ethyl sulfonyl phenylacetic acids as the key internediates through amide condensation,Suzuki reaction as the key steps to bulid different phenyl-mixed aronmatic ring sketeton.Compounds with pyrazole,thiazole,imidazole and 1,3,4-oxazole was synthesized for 4a,4b,4c,4d,4e,4f,5a and 5b.Amide-reversed compounds 6a,6b and 6c were also constructed with p-sulfonyl benzylamine and its derivatives as key intermediates.?3?The binding activities of the as-synthesized compounds with ROR?t-LBD in vitro were detected by time resolved fluorescence in GAL4 cells and h PBMC cells.The results showed that the binding activity of thiazole-pyrrole compound ethesulfonyl was equivalent to that of VTP-1,but the cell activity decreased.The sulfonyl with linear alkyl substitution had better activity than branched alkyl group.The cell activity was also decreased when the amide was changed to carbamate or urea.The binding activities of phenylacetylene compounds were close to that of VTP-1,but no activity was detected in h PBMC IL-17.In addition,the binding activity of ROR?t was not improved,The binding activity of ROR?t was also decreased whether electron absorbing or electron donor substituents were introduced into the 2 or 3 positions of benzene ring.When directly replacing propargyl with heterocyclic ring,it was found that thiazole ring had the best activity.Hydroxymethyl side chain was introduced into the CH₂position of benzyl in the head of thiazole ring,demonstrating excellent binding activity and cellular activity.Introduction of chiral hydroxymethyl suggested that R configuration was significantly superior to S configuration.Through the amide inversion strategy,compounds with better binding activity and cellular activity were also found.Finally,after a series of modifications,compounds 5a and 6b with good in vitro activity were obtained in this paper(5a,F_po?rat?=61.4%,F_po?mouse?=65.5%;6b,F_po?rat?=71.2%,F_po?mouse?=84.7%).The PK tests in rats and mice were carried out,and both compounds had good oral absorption and bioavailability to be druggable.In addition,compared with lead compound VTP-1,the exposure and half-life of oral absorption were significantly increased.In conclusion,20 compounds were designed and synthesized in this study based on ligand drug design strategy and bioelectronic isolayout,skeletal transition,conformational restriction and other drug transformation strategies.The binding and cell activities of compounds 5a and6b were found to be comparable to that of VTP-1,and the PK properties were even better,which was expected to provide a new idea for obtaining a more effective and safer novel targeted ROR?t-targeted small molecule inhibitor.