Mitigation of Oxidation in Therapeutic Antibody Formulations: a Biochemical Efficacy and Safety Evaluation of N-Acetyl-Tryptophan and L-Methionine
Mitigation of Oxidation in Therapeutic Antibody Formulations: a Biochemical Efficacy and Safety Evaluation of N-Acetyl-Tryptophan and L-Methionine
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DOI：
10.1007/s11095-018-2467-5
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发表时间：
2018-11-01
影响因子：
3.7
通讯作者：
Salisbury, Cleo M.
中科院分区：
文献类型：
--
作者：
Dion, Michelle Z.;Leiske, Danielle;Salisbury, Cleo M.
目的 生物治疗药物在制造和储存过程中可能容易氧化。已知游离的 L-蛋氨酸可以保护蛋白质中的蛋氨酸残基免遭氧化。类似地，游离色氨酸和其他吲哚衍生物已被证明可以保护色氨酸残基免遭氧化。 N-乙酰基-DL-色氨酸先前被认为是比色氨酸潜在的更好的抗氧化剂，因为它具有较低的氧化电位，并且在光照下产生较少的过氧化物。本研究旨在确认 N-乙酰基-DL-色氨酸和 L-蛋氨酸作为生物治疗药物配方成分的抗氧化功效和安全性。方法 在 N-乙酰基-DL-色氨酸和 L-蛋氨酸存在下对抗体进行 AAPH 和光照。通过肽图对相关CDR和Fc残基的氧化进行定量。进行了计算机、体外和体内研究，以评估 N-乙酰-DL-色氨酸和 L-蛋氨酸的安全性。结果 肽图分析表明，N-乙酰-DL-色氨酸可有效保护色氨酸免受 AAPH 应激，并且 N-乙酰-DL-色氨酸和 L-蛋氨酸的组合可保护色氨酸和蛋氨酸免受 AAPH 应激。安全性评估表明两种赋形剂的安全性均可接受。结论 N-乙酰色氨酸和 L-蛋氨酸可有效减少抗体中易受影响的色氨酸和蛋氨酸残基的氧化，并且可安全用于胃肠外生物治疗制剂。
Purpose Biotherapeutics can be susceptible to oxidation during manufacturing and storage. Free L-methionine is known to protect methionine residues in proteins from oxidation. Similarly, free tryptophan and other indole derivatives have been shown to protect tryptophan residues from oxidation. N-acetyl-DL-tryptophan was previously identified as a potentially superior antioxidant to tryptophan as it has a lower oxidation potential and produces less peroxide upon light exposure. This study sought to confirm the antioxidant efficacy and safety of N-acetyl-DL-tryptophan and L-methionine as formulation components for biotherapeutic drugs.Methods Antibodies were subjected to AAPH and light exposure in the presence of N-acetyl-DL-tryptophan and L-methionine. Oxidation in relevant CDR and Fc residues was quantified by peptide map. In silico, in vitro, and in vivo studies were performed to evaluate the safety of N-acetyl-DL-tryptophan and L-methionine.Results Peptide mapping demonstrated that N-acetyl-DL-tryptophan was effective at protecting tryptophans from AAPH stress, and that the combination of N-acetyl-DL-tryptophan and L-methionine protected both tryptophan and methionine from AAPH stress. The safety assessment suggested an acceptable safety profile for both excipients.Conclusions N-acetyl-tryptophan and L-methionine effectively reduce the oxidation of susceptible tryptophan and methionine residues in antibodies and are safe for use in parenteral biotherapeutic formulations.