Department of Bioengineering, University of Washington, Seattle, Washington 98195, United States.
Institute of Biomedical Sciences, Academia Sinica, Taipei 11529, Taiwan.
J Control Release. 2021 Mar 10;331:142-153. doi: 10.1016/j.jconrel.2021.01.015. Epub 2021 Jan 11.
The generation of anti-PEG antibodies in response to PEGylated proteins, peptides, and carriers significantly limits their clinical applicability. IgM antibodies mediate the clearance of these therapeutics upon repeat injection, resulting in toxicity and hindered therapeutic efficacy. We observed this phenomenon in our polymer platform, virus-inspired polymer for endosomal release (VIPER), which employs pH-sensitive triggered display of a lytic peptide, melittin, to facilitate endosomal escape. While the polymer-peptide conjugate was well tolerated after a single injection, we observed unexpected mortality upon repeat injection. Thus, the goal of this work was to enhance the safety and tolerability of VIPER for frequent dosing. Based on previous reports on anti-PEG antibodies and the adjuvant activity of melittin, we characterized the antibody response to polymer, peptide, and polymer-peptide conjugates after repeat-dosing and measured high IgM titers that bound PEG. By substituting the L-amino acid peptide for its D-amino acid enantiomer, we significantly attenuated the anti-PEG antibody generation and toxicity, permitting repeat-injections. We attempted to rescue mice from L-melittin induced toxicity by prophylactic injection of platelet activating factor (PAF) antagonist CV-6209, but observed minimal effect, suggesting that PAF is not the primary mediator of the observed hypersensitivity response. Overall, we demonstrated that the D-amino acid polymer-peptide conjugates, unlike L-amino acid polymer-peptide conjugates, exhibit good tolerability in vivo, even upon repeat administration, and do not elicit the generation of anti-PEG antibodies.
针对 PEG 化蛋白、肽和载体产生的抗聚乙二醇(PEG)抗体,极大地限制了它们的临床应用。IgM 抗体介导这些治疗药物在重复注射后的清除,导致毒性和治疗效果受阻。我们在聚合物平台(病毒启发的内体释放聚合物(VIPER))中观察到了这种现象,该平台利用 pH 敏感触发展示溶细胞肽蜂毒素,以促进内体逃逸。虽然聚合物-肽缀合物在单次注射后具有良好的耐受性,但我们在重复注射时观察到了意外的死亡率。因此,这项工作的目标是提高 VIPER 的安全性和耐受性,以实现频繁给药。基于先前关于抗聚乙二醇抗体和蜂毒素佐剂活性的报道,我们在重复给药后对聚合物、肽和聚合物-肽缀合物的抗体反应进行了表征,并测量了与 PEG 结合的高 IgM 滴度。通过用其 D-氨基酸对映体取代 L-氨基酸肽,我们显著减弱了抗聚乙二醇抗体的产生和毒性,从而允许重复注射。我们试图通过预防性注射血小板激活因子(PAF)拮抗剂 CV-6209 来挽救由 L-蜂毒素引起的毒性,但观察到最小的效果,表明 PAF 不是观察到的过敏反应的主要介导物。总体而言,我们证明与 L-氨基酸聚合物-肽缀合物相比,D-氨基酸聚合物-肽缀合物即使在重复给药时,在体内也具有良好的耐受性,并且不会引起抗聚乙二醇抗体的产生。