Design,Synthesis And Biological Evaluation Of 1-(3,4-dichlorobenzyl)indoline-2,3-dione Derivatives In Mantle Cell Lymphoma

Mantle cell lymphoma is a rare,non-Hodgkin’s lymphoma subtype with a high degree of malignancy and poor prognosis.It has unique clinical,biological and molecular characteristics.In sidious onset of the disease,the rapid progress,and other non-Hodgkin’s lymphoma is different,most patients have been in clinical stage III to IV,and accompanied by extensive extranodal infiltration.Most patients with MCL will relapse after first-line treatment.The median overall survival after relapse is about 1-2 years,so a new treatment plan for MCL is urgently needed.Currently,drugs used for the targeted treatment of mantle cell lymphoma in clinic include proteasome inhibitors,BTK inhibitors,PI3K/AKT/mTOR pathway inhibitors,immunomodulators,drugs targeting the cell cycle,and targeted death proteins.These drugs show a certain therapeutic effect in the clinic,but gradually develop drug resistance.Many chemotherapeutic drugs use it as an anti-cancer drug to activate apoptosis.Therefore,the ability to restore apoptosis through the use of small-molecule drugs may have important therapeutic implications.Through literature investigations,we found that isatin derivative apoptosis activator 2 can strongly induce caspase-3 activation,PARP cleavage,and DNA fragmentation,thereby causing cell apoptosis.Apoptosis activator 2 is very sensitive to lymphoid cell lineages with IC50 values between 4 and 9 μM.Also,the experimental results show that it does not show strong induction of apoptosis on normal cells.In combination with the more prominent biological activity of the compound isatin derivative Tii-6p,which was found in a previous laboratory study,its growth inhibitory activity against mantle cell lymphoma was about 100 times stronger than that of Ibrutinib.Based on the structural drug design,a series of novel 1-(3,4-dichlorobenzyl)indoline-2,3-dione were designed based on the principle of splicing of reactive groups using the apoptotic activator 2 as the backbone.This study mainly investigated the effect of five amide substituents on the biological activity of mantle cell lymphoma.A total of 30 such compounds were designed and synthesized.In addition,in order to investigate the effect of the benzyl group on the activity of the compound,3,4-dichlorobenzyl was replaced,and 6 compounds of the Q series and 5 compounds of the D series were designed and synthesized.We use isatin as the starting material,through the nitration reaction,ketal protection,nitro reduction,nucleophilic substitution,amide condensation,and finally deprotected under acidic conditions to obtain the target compound K.The target compound D was prepared from nitro reduction,amide condensation,nucleophilic substitution using intermediate 3 as the starting material,and finally deprotected under acidic conditions.All compound structures have been confirmed by 1H and 13C spectra,and some compounds have been HRMS confirmed.Through literature review and Scifinder structure query,all compounds are new compounds.By evaluating the growth inhibitory activity of mantle cell lymphoma,the IC50 of the target compound against several mantle cell lymphoma cell lines was approximately 0.4 to 1.5 μM.Among them,most of the target compounds of the K and Q series had better growth inhibitory activity against mantle cell lymphoma than the positive control drug ibrutinib and the lead compound Apoptosis Activator 2.In addition,such compounds show strong growth inhibitory activity against ibrutinib-resistant cell lines.However,the Dseries compounds have weaker activity and even no activity.Combined with the results of the growth inhibitory activity on mantle cell lymphoma cell lines,a preliminary structure-activity relationship was discussed for the synthesized indoline-2,3-dione compounds.In addition,we conducted further apoptosis experiments and cell cycle arrest experiments on compounds K7 and K20.The results of apoptosis experiments showed that compounds K7 and K20 showed dose-dependent induction of apoptosis in both Rec-1 and Z138 cell lines.Cell cycle arrest experiments showed that compounds K7 and K20 treatment of Rec-1 and Z138 cell lines for 24 hours can block cell cycle in different degrees,effectively block the cell cycle and promote cell death.At present,the target and mechanism of action of such compounds are not yet clear,and the research on the mechanism has not yet been carried out in this paper.In addition,the cytotoxicity selective experiment will also be studied in later work.