基于高低亲和力抗犬PD-L1抗体与肿瘤微环境响应性连接子的抗体偶联药物制备及初步性能评估OA
Preparation and preliminary evaluation of antibody-drug conjugates targeting canine PD-L1 using high/low affinity anti-PD-L1 antibodies and tumor microenvironment-responsive linkers
免疫检查点阻断疗法在人类肿瘤治疗中取得了突破性进展,然而犬类肿瘤免疫治疗药物的开发仍面临重大挑战.随着宠物市场的崛起以及宠物医疗技术的发展,针对犬类肿瘤的抗体偶联药物(ADC)的需求愈加迫切.然而,目前针对犬类程序性死亡受体配体1(PD-L1)靶点的ADC研究尚处于起步阶段,耐药性和疗效不足问题亟待进一步探索,特别是不同抗体类型及药物抗体偶联比(DAR)对新型ADC的影响.本研究首次制备了两种靶向犬PD-L1的新型ADC,采用高/低亲和力抗犬类PD-L1抗体(A单抗与T单抗)、肿瘤微环境响应型连接子以及新一代拓扑异构酶I抑制剂Dxd,并通过控制偶联参数,实现了不同DAR值ADCs的制备.经过初步鉴定后,本研究对不同DAR值的两种ADCs进行了初步表征、稳定性分析、结合活性及细胞杀伤性能评估.结果显示,两种抗体ADCs各DAR值控制良好,均具有良好的成药性,DAR值和抗体类型对于ADCs的稳定性和活性均具有一定影响,高低亲和力抗体均具有一定的杀伤性能,且随着DAR值增加,杀伤性能增强.本研究为犬类肿瘤PD-L1 ADC的开发打下了基础,并为后续的犬源化ADC开发提供了候选药物.
Immune checkpoint blockade therapies have made groundbreaking progress in human cancer treatment,but the development of immunotherapeutic drugs for canine tumors remains a significant chal-lenge.With the rise of the pet market and advancements in veterinary medical technology,the demand for antibody-drug conjugates(ADCs)targeting canine tumors is growing.However,research on ADCs targe-ting canine Programmed Death-Ligand 1(PD-L1)is still in its infancy,with issues such as drug resistance and insufficient therapeutic efficacy requiring further exploration,particularly the impact of different anti-body types and drug-to-antibody ratio(DAR)on novel ADC performance.This study pioneers the prepa-ration of two novel canine PD-L1-targeting ADCs by integrating high/low-affinity anti-canine PD-L1 mon-oclonal antibodies(A-mAb and T-mAb),tumor microenvironment-responsive linkers,and the next-gen-eration topoisomerase I inhibitor Dxd.Through controlled conjugation parameters,ADCs with distinct DAR values were successfully prepared.Subsequent characterization,stability analysis,binding activity,and cytotoxicity evaluation revealed that both ADCs exhibited well-controlled DAR values and favorable developability.DAR values and antibody types influenced ADC stability and activity,while both high-and low-affinity antibodies demonstrated dose-dependent cytotoxicity that intensified with increasing DAR val-ues.This study lays the foundation for developing PD-L1-targeting ADCs for canine tumors and provides candidate drugs for future caninized ADC development.
PAN Zhiyuan;WANG Chenguang;QIAN Weizhu;HOU Sheng;JI Lusha;LI Jun;SUN Ziqiao;WANG Xuekun;XIE Lei;ZHANG Dapeng;GUO Huaizu;REN Yule;GUO Qingcheng;XU Jin;ZHU Weifan;ZHUANG Huangzhen;GAO Tianyu;CHEN Yi;GUO Menghui;LIU Tao
State Key Laboratory of Macromolecular Drugs and Large-scale Preparation,School of Pharmaceutical Sciences and Food Engineering,Liaocheng University,Liaocheng 252059,China||NMPA Key Laboratory for Quality Control of Therapeutic Monoclonal Antibodies,Shanghai 201203,ChinaState Key Laboratory of Macromolecular Drugs and Large-scale Preparation,School of Pharmaceutical Sciences and Food Engineering,Liaocheng University,Liaocheng 252059,China||NMPA Key Laboratory for Quality Control of Therapeutic Monoclonal Antibodies,Shanghai 201203,China||State Key Laboratory of Macromolecular Drugs and Large-scale Preparation,School of Pharmaceutical Sciences,Wenzhou Medical University,Wenzhou 325035,China||Taizhou Mabtech Pharmaceuticals Co.,Ltd.,Taizhou 225316,ChinaState Key Laboratory of Macromolecular Drugs and Large-scale Preparation,School of Pharmaceutical Sciences and Food Engineering,Liaocheng University,Liaocheng 252059,China||NMPA Key Laboratory for Quality Control of Therapeutic Monoclonal Antibodies,Shanghai 201203,China||State Key Laboratory of Macromolecular Drugs and Large-scale Preparation,School of Pharmaceutical Sciences,Wenzhou Medical University,Wenzhou 325035,ChinaState Key Laboratory of Macromolecular Drugs and Large-scale Preparation,School of Pharmaceutical Sciences and Food Engineering,Liaocheng University,Liaocheng 252059,China||NMPA Key Laboratory for Quality Control of Therapeutic Monoclonal Antibodies,Shanghai 201203,China||State Key Laboratory of Macromolecular Drugs and Large-scale Preparation,School of Pharmaceutical Sciences,Wenzhou Medical University,Wenzhou 325035,ChinaState Key Laboratory of Macromolecular Drugs and Large-scale Preparation,School of Pharmaceutical Sciences and Food Engineering,Liaocheng University,Liaocheng 252059,China||NMPA Key Laboratory for Quality Control of Therapeutic Monoclonal Antibodies,Shanghai 201203,China||State Key Laboratory of Macromolecular Drugs and Large-scale Preparation,School of Pharmaceutical Sciences,Wenzhou Medical University,Wenzhou 325035,ChinaState Key Laboratory of Macromolecular Drugs and Large-scale Preparation,School of Pharmaceutical Sciences and Food Engineering,Liaocheng University,Liaocheng 252059,China||NMPA Key Laboratory for Quality Control of Therapeutic Monoclonal Antibodies,Shanghai 201203,China||State Key Laboratory of Macromolecular Drugs and Large-scale Preparation,School of Pharmaceutical Sciences,Wenzhou Medical University,Wenzhou 325035,ChinaState Key Laboratory of Macromolecular Drugs and Large-scale Preparation,School of Pharmaceutical Sciences and Food Engineering,Liaocheng University,Liaocheng 252059,China||NMPA Key Laboratory for Quality Control of Therapeutic Monoclonal Antibodies,Shanghai 201203,China||State Key Laboratory of Macromolecular Drugs and Large-scale Preparation,School of Pharmaceutical Sciences,Wenzhou Medical University,Wenzhou 325035,ChinaState Key Laboratory of Macromolecular Drugs and Large-scale Preparation,School of Pharmaceutical Sciences and Food Engineering,Liaocheng University,Liaocheng 252059,China||NMPA Key Laboratory for Quality Control of Therapeutic Monoclonal Antibodies,Shanghai 201203,China||State Key Laboratory of Macromolecular Drugs and Large-scale Preparation,School of Pharmaceutical Sciences,Wenzhou Medical University,Wenzhou 325035,China||State Key Laboratory of Macromolecular Drugs and Large-scale Preparation,Shanghai Zhangjiang Biotechnology Co.,Ltd.,Shanghai 201203,ChinaState Key Laboratory of Macromolecular Drugs and Large-scale Preparation,School of Pharmaceutical Sciences and Food Engineering,Liaocheng University,Liaocheng 252059,China||NMPA Key Laboratory for Quality Control of Therapeutic Monoclonal Antibodies,Shanghai 201203,China||State Key Laboratory of Macromolecular Drugs and Large-scale Preparation,School of Pharmaceutical Sciences,Wenzhou Medical University,Wenzhou 325035,ChinaState Key Laboratory of Macromolecular Drugs and Large-scale Preparation,School of Pharmaceutical Sciences and Food Engineering,Liaocheng University,Liaocheng 252059,China||NMPA Key Laboratory for Quality Control of Therapeutic Monoclonal Antibodies,Shanghai 201203,China||State Key Laboratory of Macromolecular Drugs and Large-scale Preparation,School of Pharmaceutical Sciences,Wenzhou Medical University,Wenzhou 325035,ChinaState Key Laboratory of Macromolecular Drugs and Large-scale Preparation,School of Pharmaceutical Sciences and Food Engineering,Liaocheng University,Liaocheng 252059,China||NMPA Key Laboratory for Quality Control of Therapeutic Monoclonal Antibodies,Shanghai 201203,China||State Key Laboratory of Macromolecular Drugs and Large-scale Preparation,School of Pharmaceutical Sciences,Wenzhou Medical University,Wenzhou 325035,China||State Key Laboratory of Macromolecular Drugs and Large-scale Preparation,Shanghai Zhangjiang Biotechnology Co.,Ltd.,Shanghai 201203,ChinaState Key Laboratory of Macromolecular Drugs and Large-scale Preparation,School of Pharmaceutical Sciences and Food Engineering,Liaocheng University,Liaocheng 252059,China||NMPA Key Laboratory for Quality Control of Therapeutic Monoclonal Antibodies,Shanghai 201203,ChinaState Key Laboratory of Macromolecular Drugs and Large-scale Preparation,School of Pharmaceutical Sciences and Food Engineering,Liaocheng University,Liaocheng 252059,China||NMPA Key Laboratory for Quality Control of Therapeutic Monoclonal Antibodies,Shanghai 201203,China||State Key Laboratory of Macromolecular Drugs and Large-scale Preparation,School of Pharmaceutical Sciences,Wenzhou Medical University,Wenzhou 325035,China||Taizhou Mabtech Pharmaceuticals Co.,Ltd.,Taizhou 225316,ChinaState Key Laboratory of Macromolecular Drugs and Large-scale Preparation,School of Pharmaceutical Sciences and Food Engineering,Liaocheng University,Liaocheng 252059,China||NMPA Key Laboratory for Quality Control of Therapeutic Monoclonal Antibodies,Shanghai 201203,China||State Key Laboratory of Macromolecular Drugs and Large-scale Preparation,School of Pharmaceutical Sciences,Wenzhou Medical University,Wenzhou 325035,ChinaState Key Laboratory of Macromolecular Drugs and Large-scale Preparation,School of Pharmaceutical Sciences and Food Engineering,Liaocheng University,Liaocheng 252059,China||NMPA Key Laboratory for Quality Control of Therapeutic Monoclonal Antibodies,Shanghai 201203,ChinaState Key Laboratory of Macromolecular Drugs and Large-scale Preparation,School of Pharmaceutical Sciences and Food Engineering,Liaocheng University,Liaocheng 252059,China||NMPA Key Laboratory for Quality Control of Therapeutic Monoclonal Antibodies,Shanghai 201203,ChinaState Key Laboratory of Macromolecular Drugs and Large-scale Preparation,School of Pharmaceutical Sciences and Food Engineering,Liaocheng University,Liaocheng 252059,China||NMPA Key Laboratory for Quality Control of Therapeutic Monoclonal Antibodies,Shanghai 201203,ChinaState Key Laboratory of Macromolecular Drugs and Large-scale Preparation,School of Pharmaceutical Sciences and Food Engineering,Liaocheng University,Liaocheng 252059,China||NMPA Key Laboratory for Quality Control of Therapeutic Monoclonal Antibodies,Shanghai 201203,ChinaState Key Laboratory of Macromolecular Drugs and Large-scale Preparation,School of Pharmaceutical Sciences and Food Engineering,Liaocheng University,Liaocheng 252059,China||NMPA Key Laboratory for Quality Control of Therapeutic Monoclonal Antibodies,Shanghai 201203,ChinaState Key Laboratory of Macromolecular Drugs and Large-scale Preparation,School of Pharmaceutical Sciences and Food Engineering,Liaocheng University,Liaocheng 252059,China||NMPA Key Laboratory for Quality Control of Therapeutic Monoclonal Antibodies,Shanghai 201203,China||State Key Laboratory of Macromolecular Drugs and Large-scale Preparation,School of Pharmaceutical Sciences,Wenzhou Medical University,Wenzhou 325035,China||Department of Oncology,Huashan Hospital,Fudan University,Shanghai 200040,China
农业科技
抗体药物偶联物犬类肿瘤免疫疗法程序性死亡受体配体1药物抗体偶联比
antibody-drug conjugates(ADCs)canine tumorsimmunotherapyprogrammed death-ligand 1(PD-L1)drug-to-antibody ratio(DAR)
《聊城大学学报（自然科学版）》 2026 (1)
94-105,12
国家自然科学基金项目(22304067)上海东方英才项目(CYQN2023031)山东省自然科学基金(ZR2023QB257,ZR2023QB196)山东省高等学校"青创团队计划"团队(2022KJ111)光岳青年学者创新团队项目(LCUGYTD2022-04)资助
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