陆军军医大学学报2026，Vol.48Issue(3)：251-260,10.DOI:10.16016/j.2097-0927.202510007
新型分子胶LTL-04-061通过CRBN介导的泛素-蛋白酶体通路靶向降解OTULIN并显著抑制胃癌细胞增殖
Novel molecular glue LTL-04-061 targets OTULIN for degradation via the CRBN-mediated ubiquitin-proteasome pathway and significantly suppresses gastric cancer cell proliferation
摘要
Abstract
Objective To screen novel molecular glues targeting ovarian tumor domain deubiquitinase with linear linkage specificity(OTULIN)and provide a new therapeutic strategy for gastric cancer.Methods Gastric cancer cell lines with high OTULIN expression(AGS-NC,SNU601-NC)and OTULIN-knockout cells(AGS-sgOTULIN,SNU601-sgOTULIN)were used.Experimental groups included:control(DMSO),LTL-04-061 concentration gradients(0.1,0.3,1.0,3.0,5.0,7.0,10.0 µmol/L),and proteasome inhibitor intervention(MG-132/MLN4924+5 µmol/L LTL-04-061).OTULIN protein expression was analyzed by Western blotting,while mRNA levels were quantified by qPCR.Protein interactions were validated by co-immunoprecipitation(Co-IP).Molecular docking and 100 ns molecular dynamics(MD)simulations were performed to analyze complex conformations.Anti-proliferative effects were assessed using CCK-8,cell proliferation,and colony formation assays.Results LTL-04-061 degraded OTULIN protein in a concentration-dependent manner(0.1 to 10.0 µmol/L),with significant degradation at 5 µmol/L.At this concentration,OTULIN mRNA levels remained unchanged(P>0.05).Degradation was blocked by MG-132/MLN4924,indicating dependence on the CRBN-mediated ubiquitin-proteasome pathway.Co-IP confirmed LTL-04-061-induced OTULIN-CRBN ternary complex formation.Key interactions involved OTULIN HIS339 and CRBN TRP378/HIS380.MD simulations showed stable complex conformation after 50 ns(RMSD fluctuation<0.1 nm),sustained hydrogen bonds(80 to 100 ns),and stable radius of gyration(Rg)and solvent-accessible surface area(SASA),indicating no structural expansion/collapse.LTL-04-061 concentration-dependently suppressed gastric cancer cell proliferation and colony formation.CCK-8 assays revealed IC50 values of 8.95 µmol/L(AGS)and 11.70 µmol/L(SNU601).OTULIN knockout significantly attenuated this inhibition(P<0.01).Conclusion OTULIN exerts pro-oncogenic effects in gastric cancer.The molecular glue LTL-04-061 targets OTULIN for CRBN-dependent ubiquitin-proteasomal degradation,thereby suppressing malignant proliferation.This identifies LTL-04-061 as a promising precursor for molecularly targeted gastric cancer therapy.关键词
胃癌/OTULIN/分子胶/LTL-04-061/靶向治疗Key words
gastric cancer/OTULIN/molecular glue/LTL-04-061/targeted therapy分类
医药卫生引用本文复制引用
刘振昆,彭紫依,向俊宇,王斌,蒋白山,邱秋,李玉红..新型分子胶LTL-04-061通过CRBN介导的泛素-蛋白酶体通路靶向降解OTULIN并显著抑制胃癌细胞增殖[J].陆军军医大学学报,2026,48(3):251-260,10.基金项目
国家重点研发计划项目(2023YFC3402102) （2023YFC3402102）
重庆市科卫联合医学科研项目(2020FYYX028) （2020FYYX028）
合川人医发(2024)109号邱秋登峰人才项目 Supported by the National Key Research and Development Program of China(2023YFC3402102),the Joint Medical Research Project of Chongqing Science and Technology Commission and Chongqing Health Commission(2020FYYX028),and the Qiu Qiu Dengfeng Talent Project(2024-109)of Hechuan People's Hospital of Chongqing. （2024）
