Design,Synthesis And Anti-GBM Activity Evaluation Of PTPRζ Allosteric Inhibitors And Study On The Synthesis Of Key Intermediates Of Oral Anti-COVID-19 Drug Ensitrelvir

This Master’s thesis contains two parts.This Master’s thesis contains two parts.Part I:Design,Synthesis and Anti-GBM Activity Evaluation of PTPRζAllosteric InhibitorsObjective:NAZ2329 is the first cell permeable PTPRζallosteric inhibitors,which can cause PTPRζThe catalytic effect of WPD ring structure changes,causing it to lose catalytic activity.And it can inhibit the proliferation and migration of GBM cells in vitro and in vivo.However,it has disadvantages such as low activity and poor blood-brain barrier permeability.In order to further improve the anti-GBM activity of NAZ2329,this thesis proposes to structurally modify it,obtain derivatives and test the anti-GBM activity in vitro.The structure-activity relationship was summarized to guide the subsequent design and synthesis of PTPRζallosteric inhibitors with NAZ2329 as the lead compound.Methods:（1）Design of a series of PTPRζinhibitors by visualization and analysis of the protein crystal complex map of PTPRζand NAZ239;（2）Improvement of the synthetic route of the lead compound NAZ2329 using inverse synthetic analysis and efficient synthesis of the designed series of compounds using the new improved synthetic route;（3）The synthesized compounds were tested for proliferation inhibitory activity on human-derived U251-MG cells using the Cell Titer-Glo method.Results:（1）The design of a series of PTPRζinhibitors modified for the Ar¹,Ar²,X-linked atoms and the four Ar³parts of the precursor NAZ2329;（2）Improved synthetic route for the lead compound NAZ2329,increasing the yield of the key reaction step from 0.94%to 34.1%,and the synthesis of 29 new compounds using an improved strategy with structural confirmation by NMR and mass spectrometry;（3）The compound GMC-27 showed the best in vitro anti-GBM activity(IC₅₀=12.7μM),which was approximately 4-fold better than the lead NAZ2329（56.7μM）.Conclusion:（1）An improved synthetic route to NAZ2329 suitable for medicinal chemistry studies was obtained;（2）Preliminary conformational relationships are summarised as follows:for the Ar¹part,activity is best when1-hexyloxy-4-trifluoromethylbenzene is used,and the molecular volume of this part should not be too large;for the Ar²part,an ortho-substituted benzene ring is preferred,with a loss of activity due to substitution of the para-substituted benzene ring;for the X heteroatom part,S is preferred,with a reduction in activity due to the use of O substitution;for the Ar³part,S-1-phenylethylamine is preferred,with a reduction in activity due to the use of carbamides or directly linked aromatic amines.The preliminary summary of the conformational relationships provides a theoretical basis for the subsequent design and synthesis of PTPRζmetamorphism inhibitors based on NAZ2329 as the lead compound.Part II:Study on the Synthesis of Key Intermediates of Oral Anti COVID-19 Drug EnsitrelvirObjective:Ensitrelvir is the first orally administered non-covalent,non-peptide novel coronavirus 3CLprotease（SARS-Co V-2 3CLprotease）inhibitor with a low yield of the key intermediate triazinone-triazole in the original synthetic strategy.In order to improve the yield of the key intermediate triazinone-triazole,this paper proposes to investigate the synthesis strategy of this key intermediate.Methods:Based on the concept of green chemistry,the synthetic strategy of triazolium derivatives and Ensitrelvir key intermediates,triazinone-triazolium,was optimized by means of inverse synthetic analysis and one-pot synthesis strategy.Results:（1）Synthesis of 3-（chloromethyl）-1,2,4-triazole in 72%yield from green,inexpensive 2-chloroacetamide using a one-pot tandem reaction（condensation and cyclization reaction）;（2）The N-alkylation reaction and the highly selective N¹-methylation reaction were achieved by a one-pot tandem reaction to give Ensitrelvir the key intermediate triazinone-triazole in 54%yield;（3）Two major process impurities were synthesized and identified;the N³-substituted triazinone-triazole isomer impurity was rarely produced during the reaction and the triazinone methyl impurity could be removed by post-processing purification;（4）The four intermediates containing the triazole ring were found to be stable up to 100 ^oC by DSC,allowing for a safe reaction and post-treatment process.Conclusion:（1）A synthetic strategy developed for the synthesis of3-（chloromethyl）-1,2,4-triazoles by a one-pot tandem reaction（condensation and cyclization reactions）,which avoids the use of toxic feedstocks and chlorinated reagents and improves atomic economy compared to existing synthetic routes;（2）Another one-pot tandem reaction（N-alkylation reaction and highly selective N¹-methylation）was developed to synthesise Ensitrelvir’s key intermediate,triazinone-triazole,with a small increase in yield（54%）compared to the original synthetic route（45%）.The green and efficient two-pot synthesis strategy developed opens up new avenues for further research and development of Ensitrelvir analogues.