Design,Synthesis And Activity Study Of GPR40 Agonists And MMP-9 Inhibitors With Chiral Sulfoxide Group

1.Design,Synthesis and Activity Study of GPR40 Agonists with Chiral Sulfoxide GroupBackgroundDiabetes mellitusis a major metabolic disease affecting human health,and there are more than 90%diabetic who have type 2 diabetes.With growing needs for novel antidiabetic drugs,GPR40 has been developed as a novel and potential therapeutic target for type 2 diabetes.According to the mechanism of action,the iinsulinotropic effects via GPR40 activation are dependent on glucose concentration,therefore,a selective GPR40 agonist has the advantage of low risk of hypoglycemia over traditional antidiabetic agents(e.g.sulfa hypoglycemic drugs).Thus GPR40 has become an attractive target for lowering blood sugar levels in type 2 diabetes.Design,synthesis and activity evaluation of target compoundsIn recent years,a variety of potent GPR40 agonists have been reported in the literatures.There are some promising small molecules at different stages of drug research and development.TAK-875,or fasiglifam,is an orally available.potent and selective partial agonist of GPR40,which reached phase III clinical trials for the treatment of type 2 diabetes,unfortunately,it was discontinued from clinical trials due to suspected liver toxicity.In the study,according to bioisosterism principles.switching sulfonyl of TAK-875 to sulfinyl changed the parent structure minimally to keep the favourable activity.Generally,the polarity of sulfoxide compounds is greater than sulfone compounds in the homologous series,hence,the water solubility of the sulfoxide analogues of TAK-875 may be increased.Meanwhile,the lipotoxicity will be reduced due to low lipid solubility.Pharmacokinetics may be improved,and in vivo activity would be much better.The chiral sulfoxide analogues which was introduced by modifying the sulfone group of TAK-875 should be respectively investigated drug-likeness.Firstly,TAK-875,compound g19 was synthesizedas a control drug.Secondly,the racemate sulfoxide compound(g24)was synthesized originating the appropriate intermediate from TAK-875,followed by nucleophilic substitution,reduction reaction,Mitsunobu reaction,oxidation and hydrolysis.Thirdly,preparative chiral HPLC separation of g24 using a CHIRALPAK ADH column afforded epimeric g25(S,S,100.0%de)and g26(R,S,100.0%de).At last,asymmetric oxidation synthesized abundant sulfoxide analogues with high optical purity.The polarity and water solubility of target compounds were tested.The EC50 values for the GPR40 agonists were determinated by calcium flux primary assay.Oral glucose tolerance test(OGTT)was carried out on rat diabetic model.The target compounds also estimated pharmacokinetics and hepatotoxicity in vitro and in vivo to investigate the drug-like property.ResultsIn this research,intermediates and target compounds were obtained and identified by IR,1H NMR,1C NMR,ESI-MS and HR-MS.Optical rotation was measured on a MCP200 polarimeter.The optical purity was determined with an analytical chiral HPLC CHIRALPAK IA column.The absolute configuration was determined by literatures or circular dichroism spectra analysis.Literature retrieval proved that all the target compounds were new and not reported.Our results showed that the GPR40 agonistic activities of these sulfoxide and sulphone analogs were comparable.g29(main g25)exhibited more potent in vivo glucose-lowering effect than TAK-875.In vivo pharmacokinetic research revealed that g25(S,S)could be metabolized to g19 more easily than its epimer g26(R,S).What's more,g25(S,S)exhibited better pharmacokinetic profiles characterized by better oral bioavailability and longer period of effectiveness.After oral administration for one month,blood biochemical features were examined on the diabetic rats,the index of glutamic-pyruvic transaminase and glutamic oxalacetic transaminase in g25(S,S)group was higher than control group.g25(S,S)induced HL7702 cell apoptosis more potent than g19.ConclusionThere is almost no effect on activity in vitro,when transforming sulfonyl of TAK-875 to sulfinyl,which can be explained by the structure of hGPR40-TAK-875 co-complex.Meanwhile,the water solubility of the sulfoxide analogues is improved.g25(S,S)has better pharmacodynamics and pharmacokinetics than TAK-875,and it was easily metabolized to TAK-875 in vivo,therefore it mightbe have hepatotoxicity which was confirmed by blood biochemical examination in type 2 diebetes rats mode.g25(S,S)induced HL7702 cell apoptosis indicates itself has hepatotoxicity.Therefore,designing GPR40 agonist should be considered hepatotoxicity carefully.And the drug design of switching sulfonyl to sulfinyl is a "double-edged sword",replacing with bioisosterism depends on the drugability of parent compound seriously,when the parent compound has favourable druggability or marketed drug,research and development of the sulfoxide derivatives maybe successful,however,if the parent compound was failed,derivatives will have almost no opportunity to success,2.Design,Synthesis and Activity Study of MMP-9 Inhibitors with Chiral Sulfoxide GroupBackgroundA complication of diabetes is the inability of wounds to heal in diabetic patients.There are no readily available methods for intervention of this disease.Diabetic wounds are refractory to healing due to the involvement of activated MMP-9,which remodel the tissue resulting in apoptosis.The treatment of diabetic wounds with a selective MMP-9 inhibitor holds great promise in providing heretofore-unavailable opportunities for intervention of this disease.Design,synthesis and activity evaluation of target compoundsND-322 and ND-364 with high MMP-9 inhibitory activity have been reported in the literatures.But it should be mentioned that both of them are racemates.Herein,in order to investigate the effects of configuration of thiirane moiety on MMP-9 inhibition,the optically active(R)-and(S)-enantiomers of compounds ND-322 and ND-364 were synthesized and evaluated.ND-364 which was N-acetylated metabolite from ND-322 in vivo was even more potent.Sulfonyl group of ND-364 will be transformed to sulfinyl to find novel chiral sulfoxide MMP-9 inhibitors.Moreover,anilino group will be protected with(Boc)2O to investigate the effects of the size of substituent group.Synthesis commenced with chlorosulfonation of optically pure glycidol as chiral source to get intermediate,which was then reacted with 4-mercaptophenol,the stereocenter was not scrambled during this step.Formation of the epoxide ring and reacting with 1-fluoro-4-nitrobenzene took place in one step in the presence of C22CO2.The nitro was reduced by zinc powder,then fitered by kieselguhr with the filtrate without further purification being added(Boc)2O for protecting amino group or acetylization.Followed by sulfide oxidation,thiirane ring formation and N-Boc deprotection to obtain the title compounds.The step,conversion of the oxirane to the thiirane ring,involved inversion of stereochemistry.Moreover,different configurations of sulfoxide analogues were got by asymmetric oxidation,then treated with thiourea to generate the title compounds.The inhibitory activity of MMP-2,MMP-9,APN and HDACs was tested for the title compounds in vitro.ResultsIn this research,the intermediates and target compounds were obtained and identified by IR,1H NMR,ESI-MS and HR-MS.Optical rotations was measured on a MCP200 polarimeter.The optical purity was determined with an analytical chiral HPLC CHIRALPAK IA column.The absolute configuration was determined by literatures and circular dichroism spectra analysis.Literature retrieval proved that all the target compounds were new and not reported.na8?nb8?nall and nbll were selective inhibitors of MMP-2 and MMP-9.Compared to ND-322 and ND-364,they kept activity respectively.ConclusionThe configuration of thiirane moiety has little effect on inhibitory activity.And transforming sulfonyl to sulfinyl reduced inhibitory activity obviously,according to the unique mechanism of action,there should be strong electron-withdrawing group at the position,which is favorable for improving activity.Because of steric hindrance,compounds with too large group connecting with aniline will lose the inhibitory activity.This research would provide impotant reference value for the development of novel highly potent selective inhibition against MMP-9.