Design, Synthesis And Evaluation Of Antitumor Activity Of CRM1 Inhibitors

CRM1(Chromosome region maintenance 1),also known as exportin 1 or XPO1.It is a eukaryotic protein that mediates the nucleocytoplasmic trafficking of nuclear protein,r RNA,sn RNA and m RNA.The exportin CRM1 exports most of the tumor suppressor proteins and oncoproteins.And abnormal expression of CRM1 is closely related to the tumors accruing,development and drug-resistant.Thus CRM1 has become an important target for anticancer drug development.By literature review,we found that most CRM1 inhibitors have the α,β-unsaturated δ-lactone unit including Goniothalamus,Leptomycin B and other compounds.The preliminary research proves that the modification at the unsaturated double bonds of target compounds would lose their antitumor biological activity and the different side chains in the compounds significantly influence the activity.Based on the previous researches and the basic structure of control 2 and Goniothalamus which were used as the lead compounds,their structural modification and structure-activity relationship are implemented in order to develop new CRM1 inhibitors.The main work is as follows:1.Synthesis and bioactivity of 6-oxy/amino/thiomethyl-5,6-dihydro-2H-pyran-2-oneUsing control 2 as the lead compound,we retained unsaturated lactone ring,modified side chains and brought in the different heteroatom(oxygen,nitrogen,sulfur)in the 6 position of the side chains.we can get 14 oxymethyl compounds,4 aminomethyl compounds,1 thiomethyl compound and 1 sulfinylmethyl compound through substitution,ring-opening,esterification,oxidation,cyclization reaction.We evaluated their antitumor activity and results indicate that the IC50 values are greater than or equal to 2.31μM against the tumor cells.The oxymethyl compounds with the electron-withdrawing group on benzene ring,the aminomethyl compounds,and the thiomethyl compound attenuate or eliminate their antitumor activity.However,the other oxymethyl compounds with electron-donating group on benzene ring and the sulfinylmethyl compound have good antitumor.the electron-withdrawing group on benzene ring and 6-heteroatom have a significant impact on the antitumor activity.2.Synthesis and bioactivity of 6-aminoethyl-5,6-dihydro-2H-pyran-2-oneBased on the good antitumor activity of the sulfinylmethyl compound,we study the impact of the amide groups on the antitumor activity through the acylation of amino.Method 17 the 6-aminoethyl compounds were synthesized via acylation,cyclization,thionation reaction.The antitumor activity was evalueated and the results indicate that most compounds lose their biological activity.The the amide groups of the side chains may have a significant impact on the antitumor activity.3.The effect of the promising candidate on cell cycle distribution,the apoptosis rate and CRM1 expression of Hela cells.CRM1 is highly expression in the Hela cells.Consulting the previous research,we selected the compound I11 with the best antitumor activity and studied its influence on cell cycle distribution and the apoptosis rate of Hela cells.The results show that the apoptosis rate of Hela cells was up to 22.7% and the cell cycle progression was arrested in G1 phase in the Hela cells at the 20μM concentration.And the level of CRM1 protein expression became markedly reduced in a dose-dependent manner on treatment with the compound I11 by Western blotting.In conclusion,based on the basic structure of Goniothalamus and the potential CRM1 inhibitor,their structural modification and structure-activity relationship are researched.A total of 37 target compounds were synthesized,and their structures were all confirmed by 1H NMR,13 C NMR and HRMS.The obtained compounds were not reported before.Among them,the compound I11 has better antitumor activity against MGC-803 cell lines with the IC50 values of 2 μM,Hela cell lines with the IC50 values of 4 μM and H1299 cell lines with the IC50 values of 6 μM compared with control 2 in vitro;the compound I8 has better antitumor activity against MGC-803 cell lines with the IC50 values of 8 μM and Hela cell lines with the IC50 values of 8 μM compared with control 2 in vitro.The compound I11 could make Hela cells stop in the period of G1 and lead to apoptosis.And the level of CRM1 protein expression became markedly reduced in a dosedependent manner on treatment with the compound I11.The results of these studies have laid a good foundation for the new CRM1 inhibitor.