新型Bcl-2小分子抑制剂的设计、合成与初步活性评价

Design,synthesis and preliminary activity evaluation of novel Bcl-2 protein inhibitors

目的设计并合成一系列新型Bcl-2小分子抑制剂,测试其对Bcl-2和Bcl-2 G101V(Bcl-2突变体)与BH3-only蛋白相互作用的抑制活性,初步探究其构效关系,为后续相关研究提供参考。方法以临床化合物BGB-11417为先导化合物,通过骨架跃迁等方法,设计新型含螺环结构的Bcl-2小分子抑制剂。以苯甲醛为原料,通过取代、环化和偶联等反应合成目标化合物,并通过1H NMR和LC-MS进行结构确定。采用时间分辨荧光共振能量转移(TR-FRET)评价目标化合物对Bcl-2和Bcl-2 G101V(Bcl-2突变体)与BH3-only蛋白相互作用的抑制能力。结果共合成8个新型螺环Bcl-2小分子抑制剂,其中29a[IC_(50)(Bcl-2):0.8 nmol·L^(-1),IC_(50)(Bcl-2 G101V):55.41 nmol·L^(-1)],29d[IC_(50)(Bcl-2):0.27 nmol·L^(-1),IC_(50)(Bcl-2 G101V):18.65 nmol·L^(-1)]对Bcl-2和Bcl-2 G101V与BH3-only蛋白的相互作用有较好的抑制活性。结论建立了一种新型螺环Bcl-2小分子抑制剂合成方法,发现了对Bcl-2和Bcl-2 G101V(Bcl-2突变体)与BH3-only蛋白相互作用有较好抑制活性的新化合物,其中29d具有进一步研究的价值。

Objective To design and synthesize a series of novel small molecule inhibitors of Bcl-2 and test their effects on Bcl-2,Bcl-2 G101V(Bcl-2 mutant)and BH3-only protein interactions,and initially investigate their constitutive relationship,which will provide reference for subsequent related studies.Methods The clinical compound BGB-11417 was used as the lead compound,a novel small molecule inhibitor of Bcl-2 containing a spirocyclic structure was designed by scaffold hopping and other methods.The target compounds were synthesized from benzaldehyde by substitution,cyclization and coupling reactions,and the structures were determined by 1H-NMR and LC-MS.Time-resolved fluorescence resonance energy transfer(TR-FRET)was used to evaluate the inhibitory ability of the target compounds on the interactions of Bcl-2,Bcl-2 G101V(Bcl-2 mutant)with BH3-only protein.Results A total of eight novel spiro Bcl-2 small molecule inhibitors were synthesized,among which 29a[IC_(50)(Bcl-2):0.8 nmol·L^(-1),IC_(50)(Bcl-2 G101V):55.41 nmol·L^(-1)]and 29d[IC_(50)(Bcl-2):0.27 nmol·L^(-1),IC_(50)(Bcl-2 G101V):18.65 nmol·L^(-1)]showed good inhibitory activities on the interaction of Bcl-2 and Bcl-2 G101V with BH3-only protein.Conclusion A novel method for the synthesis of spiro Bcl-2 small molecule inhibitors has been established,and new compounds with good inhibitory activity against the interaction of Bcl-2 and Bcl-2 G101V(Bcl-2 mutant)with BH3-only proteins have been found,among which 29d is valuable for further study.