Oxygen Transport to Tissue XLIII
Oxygen Transport to Tissue XLIII
复制标题
氧气输送至组织 XLIII
DOI：
10.1007/978-3-031-14190-4_48
复制
发表时间：
2022
期刊：
影响因子：
--
通讯作者：
Cooper C
中科院分区：
文献类型：
--
作者：
Cooper C
蛋白质巯基残基的PEG化是用于产生稳定的药物缀合物以增加血管保留时间的常用方法。我们最近创建了一个假定的血红蛋白为基础的氧载体，使用马来酰亚胺-PEG选择性地修饰α亚基（α Ala 19 Cys）中的单个工程半胱氨酸残基。然而，马来酰亚胺-PEG加合物通过逆迈克尔反应进行解缀合，随后与内源性血浆硫醇（如人血清白蛋白或谷胱甘肽上发现的那些）交叉缀合。在先前的研究中，单砜-PEG加合物已被证明对解偶联不太敏感。因此，我们比较了我们的马来酰亚胺-PEG Hb加合物与使用单砜PEG产生的加合物的稳定性。当在1 mM还原型谷胱甘肽、20 mg/mL人血清白蛋白或人血清存在下于37 °C下孵育7天时，相应的单砜-PEG加合物显着更稳定。在所有情况下，用单砜-PEG处理的血红蛋白保留>90%的其缀合，而马来酰亚胺-PEG显示出显著的去缀合，特别是在ImM还原型谷胱甘肽存在下，其中<70%的马来酰亚胺-PEG缀合物保持完整。虽然Hb的马来酰亚胺-PEG化似乎足以用于急性使用的氧治疗，但如果需要更长的血管滞留，则诸如单砜-PEG的试剂可能更合适。
PEGylation of protein sulfhydryl residues is a common method used to create a stable drug conjugate to enhance vascular retention times. We recently created a putative haemoglobin-based oxygen carrier using maleimide-PEG to selectively modify a single engineered cysteine residue in the α subunit (αAla19Cys). However, maleimide-PEG adducts are subject to deconjugation via retro-Michael reactions, with consequent cross-conjugation to endogenous plasma thiols such as those found on human serum albumin or glutathione. In previous studies mono-sulfone-PEG adducts have been shown to be less susceptible to deconjugation. We therefore compared the stability of our maleimide-PEG Hb adduct with one created using a mono-sulfone PEG. The corresponding mono-sulfone-PEG adduct was significantly more stable when incubated at 37 °C for 7 days in the presence of 1 mM reduced glutathione, 20 mg/mL human serum albumin, or human serum. In all cases haemoglobin treated with mono-sulfone-PEG retained >90% of its conjugation whereas maleimide-PEG showed significant deconjugation, especially in the presence of 1 mM reduced glutathione where <70% of the maleimide-PEG conjugate remained intact. Although maleimide-PEGylation of Hb seems adequate for an oxygen therapeutic intended for acute use, if longer vascular retention is required reagents such as mono-sulfone-PEG may be more appropriate.