MPLA衍生物的设计、合成及作为RBD⁃hFc新冠疫苗佐剂的活性研究  

Design,synthesis and immunological evaluation of MPLA analogs as adjuvants for RBD⁃hFc based SARS⁃CoV⁃2 vaccine

作　　者：廖攀 苏诗微 陈丽清 罗翔 廖国超 孙平华[3] 叶连宝 LIAO Pan;SU Shiwei;CHEN Liqing;LUO Xiang;LIAO Guochao;SUN Pinghua;YE Lianbao(School of Pharmacy,Guangdong Pharmaceutical University,Guangzhou 510006,China;School of Pharmaceutical Sciences,Guangzhou University of Chinese Medicine,Guangzhou 510006,China;College of Pharmacy,Jinan University,Guangzhou 510632,China)

机构地区：[1]广东药科大学药学院,广东广州510006 [2]广州中医药大学中药学院,广东广州510006 [3]暨南大学药学院,广东广州510632

出　　处：《沈阳药科大学学报》2024年第3期301-313,350,共14页Journal of Shenyang Pharmaceutical University

基　　金：2022年度广东省普通高校重点领域专项(2022ZDZX2030)。

摘　　要：目的设计合成单磷酸脂质A(monophosphoryl lipid A,MPLA)的衍生物,考察其作为新冠病毒疫苗佐剂的开发潜力及C⁃1或C⁃6′位取代基对其活性的影响。方法以商业易得的葡萄糖胺为起始原料,经糖基化、缩合、磷酰化和氢化等反应完成目标化合物的合成。以临床应用的铝佐剂为阳性对照,将所得化合物与RBD⁃hFc重组蛋白新冠疫苗合用,于第0天和第14天经肌肉注射免疫小鼠。然后,分别在初次免疫后的第14、28、42、90天采血,通过检测抗体滴度与亚型、抗体血清抑制RBD结合ACE2的能力及中和抗体水平评价了目标化合物作为佐剂的免疫活性。结果合成得到5个MPLA的衍生物,产物及中间体结构经NMR和HRMS确证;生物活性测试结果表明,化合物1具有比铝佐剂更优的佐剂活性,化合物3和4的活性与铝佐剂相当。结论MPLA衍生物具有作为新冠病毒疫苗佐剂的开发潜力;初步构效关系分析表明,将MPLA的C⁃1位羟基以甲氧基和氨基替换会降低其活性,而将其C⁃6′的羟基以氨基取代可以提高活性。Objective To investigate the potential of Salmonella minnesota monophosphoryl lipid A(MPLA)as adjuvants for RBD⁃hFc based SARS⁃CoV⁃2 vaccine and the influence of functional group at C⁃1 and C⁃6′position,herein,a series of MPLA derivatives were designed and synthesized.Methods The target com⁃pounds were synthesized using the commercially available glucosamine as a starting material through a series of reactions,including glycosylation,condensation,phosphorylation,hydrogenation and so on.The adjuvant activities of compounds 1-5 were evaluated by mixing with a subunit vaccine candidate RBD⁃hFc protein through mouse immunization on day 0 and day 14 with the clinically alum adjuvant as the positive control.Each mouse was bled on days 14,28 and 42 post⁃first immunization,the immunological activity was evalua⁃ted through the determination of IgG titer and IgG isotype,the ability of antisera to inhibit the RBD protein binding to ACE2 and neutralizing antibody level.Results Five MPLA derivatives were synthesized,and the structures of the target compounds and intermediates were characterized by NMR spectra and HRMS.The immunological evaluation showed that compound 1 exhibited better adjuvant activity than Al,and the adju⁃vant activities of compounds 3 and 4 were comparable to Al.Conclusion MPLA derivatives have the poten⁃tial to be adjuvants for the development of RBD⁃hFc based SARS⁃CoV⁃2 vaccine.Preliminary structure⁃ac⁃tivity relationship analysis of MPLA analogs shows that the functional groups at C⁃1 or C⁃6′positions affect the biological activity.The replacement of the hydroxy group at C⁃1 position by methoxy or amino group will decrease the activity,however,the hydroxy group at C⁃6′position replaced by amino group can increase the activity.

关 键 词：MPLA衍生物 佐剂 RBD⁃hFc 重组蛋白新冠疫苗 

分 类 号：R914[医药卫生—药物化学]