Synthesis Of Aromatic Indazolones As Hsp90Inhibitor Prodrugs

Hsp90inhibitors are a new kind of anti-cancer drugs at present. This kind of drugs target the Hsp (heat shock protein) in order to inhibit the function of Hsps as chaperones, thus inhibit the happening and progress of cancer. Hsp90inhibitors target a special kind of proteins, which break the old thinking that anti-cancer drugs should target genes. As a result, they are regarded to be novelty and promising, which can be used with other anti-cancer drugs.Aromatic indazolones are a new kind of Hsp90inhibitors, which are found by a famous drug research Company Senerex first time and are now in clinical trials. Recent researches have suggested aromatic indazolones have promising activity in anti-cancer activity and physiology. On these basis, in order to find a new kind of high-effective and low-toxic Hsp90inhibitors,we have synthesis11new aromatic indazolones as leading compounds,then synthesis3target moleculars as we designed basis on foreign articles. All the compounds have been characterized by1H-NMR and MS. Activity detection and optimation research have been carried out selectively. We have found out that aromatic indazolones indeed have activity. The target compound AX1319has inhibition activity towards Hsp90a.The optimation experiments suggest that2e.q. EtONa in anhydrous EtOH stirred at120degrees centigrade for16hours will give the best product yield.This dissertation include following parts:A synthesis of aromatic indazolones1synthesis of aromatic indazolones in two ways1.1Synthesis of three kinds of aromatic indazolones by cyclizing derivatized dimedones with aromatic hydrazines in acid circumstance. Methods:4-substitute aromatic diazonium salt can be prepared by4-sustitut e aromatic amine and sodium nitrite in acid cool bath. The reaction solutio n can be used in next step without other treatment. The reaction solution c ould be reduced by NaHSO3and NaOH in base circumstance to give4-sub stitute aromatic hydrazine.4-substitute aromatic hydrazine can cyclized with dimedone in acid circumstance. The ultimate compounds are4-(3,6,6-trimet hyl-4-oxo-4,5,6,7-tetrahydroindazol-1-yl)benzoicacid,4-(3-(ethoxycarbonyl)-6,6-d imethyl-4-oxo-4,5,6,7-tetrahydroindazol-l-yl) benzoic acid and4-(6,6-dimethyl -4-oxo-3-(trifluoromethy1)-4,5,6,7-tetrahyddroindazol-1-yl)benzoic acid. The structures have been characterized by1HNMR and MS.1.2Synthesis of aromatic indazolones by dimedone and hydrazonoyl chlorides in base circumstance. Methods:4-substitute aromatic diazonium salt can be prepared by4-sustitute aromatic amine and sodium nitrite in acid cool bath. Drop Ethyl2-chloro acetoacetate in EtOH into the reaction when pH=5to give4-substitute hydr azonoyl chlorides. After recrystalization from EtOH,4-substitute hydrazonoyl chlorides will react with5,5-dimedone by dehydration and cyclization.After Column chromatography, we get9ultimate compounds:4-(3,6,6-trimethyl-4-oxo-4,5,6,7-tetrahydroindazol-1-yl)benzoic acid, ethyl1-(4-bromophrnyl)-6,6-dimethyl-4-oxo-4,5,6,7-tetrahydro-1H-indazole-3-carboxylate,ethy11-(4-Sulfanil amide)-6,6-dimethyl-4-oxo-4,5,6,7-tetrahydro-1H-indazole-3-carboxylate,ethyl1-(4-fluorophenyl)-6,6-dimethyl-4-oxo-4,5,6,7-tetrahydro-1H-indazole-3-carbox ylate,ethyl6,6-dimethyl-4-oxo-1-(4-(trifluoromethyl)phenyl)-4,5,6,7-tetrahydro-1H-indazole-3-carboxylate,ethyl6,6-dimethyl-l-(4-nitropheny1)-4-oxo-4,5,6,7-tetr ahydro-1H-indazole-3-carboxylate, ethyl6,6-dimethyl-4-oxo-1-p-tolyl-4,5,6,7-tetrahydro-1H-indazole-3-carboxylate,ethyl-6,6-dimethyl-4-oxo-1-phenyl-4,5,6,7-tetrahydro-1H-indazole-3-carboxylate, ethyl1-(4-cyanophenyl)-6,6-dimeth yl-4-oxo-4,5,6,7-tetrahydro-1H-indazole-3-carboxylate.The structures have been characterized by1HNMR and MS.2Synthesis of designed Hsp90inhibitorsThe three designed Hsp90inhibitors can be prepared by aromatic indazolones with HBTU and4-amino cyclohexanol in DMF.The structures are characterized by ’HNMR and MS.B Activity detection of Hsp90inhibitors.Recently researchers have found that the mechanism of Hsp90inhibitors is to combine with ATpase activity domain of Hsp90a N-domain. On this basis, using fluorescence polarization to detect the activity by combine with Hsp90a module has emerged. In our research, we also use fluorescence polarization to detect the affinity of AX1319combine with Hsp90a.According to fluorescence polarization principles, we detect the fluorescence polarization of AX1319acting with Hsp90α in homogeneous solution. We evaluate the affinity of competitive combination by acting AX1319and Hsp90a.Results show AX1319has the affinity to combine Hsp90α. The structure can be modified to develop Hsp90inhibitors.C The process optimation of AX1319Reaction regents, temperature, time and amount of base have been explored to found out the effect on output of the reaction, which target the key reaction.Methods:Relative content of product peak were detected by Agilent1100to come to conclusion, using target peak as a comparison:2e.q. EtONa in anhydrous EtOH to stir at120degrees centigrade for16hours will give the best product yield.