A novel platform for engineering blood-brain barrier-crossing bispecific biologics
A novel platform for engineering blood-brain barrier-crossing bispecific biologics
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DOI：
10.1096/fj.14-253369
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发表时间：
2014-11-01
期刊：
影响因子：
4.8
通讯作者：
Stanimirovic, Danica B.
中科院分区：
文献类型：
--
作者：
Farrington, Graham K.;Caram-Salas, Nadia;Stanimirovic, Danica B.
血脑屏障(BBB)阻止治疗性抗体进入中枢神经系统(CNS)靶点。将治疗臂与BBB跨细胞臂相结合的双特异性抗体工程可以显著增强它们的脑递送。BBB通透性单域抗体FC5是先前通过对原始的骆驼单域抗体噬菌体展示文库进行表型淘选而分离出来的。在本研究中，FC5被设计为与人Fc结构域的单价和双价融合，以优化其作为模块化脑传递平台的作用。体外研究表明，与单价融合相比，FC5与Fc的双价融合使跨脑内皮细胞单层的跨细胞率(P-APP)增加了25%。在大鼠全身给药后，观察到FC5-与对照区域抗体-Fc融合相比，FC5-Fc的表观脑暴露(来自血清和脑脊液药代动力学曲线)增加了30倍。在Hargreaves炎性疼痛模型中，使用与FC5-Fc融合蛋白化学偶联的血脑屏障不通透性神经肽Dalargin和神经肽Y来评估全身药理学效力。与FC5的单域版本相比，Fc融合FC5的血清药代动力学得到改善，药理效力提高了60倍；与Fc融合的二价和单价FC5表现出相似的全身药理效力。这项研究表明，将FC5作为BBB载体臂模块化地掺入双功能抗体或抗体-药物结合物中，为开发具有药理活性的中枢神经系统指标的生物疗法提供了一条途径。Farrington，G.K.，Caram-Salas，N.，Haqqani，A.S.，Brunette，E.，Eldredge，J.，Pepinsky，B.，Antognetti，G.，Baumann，E.，Ding，W.，Garber，E.，酱，S.，Delaney，C.，Boileau，E.，Sisk，W.P.，Stamirovic，D.B.为设计跨越血脑屏障的双功能生物制剂提供了一个新的平台。
The blood-brain barrier (BBB) prevents the access of therapeutic antibodies to central nervous system (CNS) targets. The engineering of bispecific antibodies in which a therapeutic arm is combined with a BBB-transcytosing arm can significantly enhance their brain delivery. The BBB-permeable single-domain antibody FC5 was previously isolated by phenotypic panning of a naive llama single-domain antibody phage display library. In this study, FC5 was engineered as a mono- and bivalent fusion with the human Fc domain to optimize it as a modular brain delivery platform. In vitro studies demonstrated that the bivalent fusion of FC5 with Fc increased the rate of transcytosis (P-app) across brain endothelial monolayer by 25% compared with monovalent fusion. Up to a 30-fold enhanced apparent brain exposure (derived from serum and cerebrospinal fluid pharmacokinetic profiles) of FC5- compared with control domain antibody-Fc fusions after systemic dosing in rats was observed. Systemic pharmacological potency was evaluated in the Hargreaves model of inflammatory pain using the BBB-impermeable neuropeptides dalargin and neuropeptide Y chemically conjugated with FC5-Fc fusion proteins. Improved serum pharmacokinetics of Fc-fused FC5 contributed to a 60-fold increase in pharmacological potency compared with the single-domain version of FC5; bivalent and monovalent FC5 fusions with Fc exhibited similar systemic pharmacological potency. The study demonstrates that modular incorporation of FC5 as the BBB-carrier arm in bispecific antibodies or antibody-drug conjugates offers an avenue to develop pharmacologically active biotherapeutics for CNS indications.Farrington, G. K., Caram-Salas, N., Haqqani, A. S., Brunette, E., Eldredge, J., Pepinsky, B., Antognetti, G., Baumann, E., Ding, W., Garber, E., Jiang, S., Delaney, C., Boileau, E., Sisk, W. P., Stanimirovic, D. B. A novel platform for engineering blood-brain barrier-crossing bispecific biologics.