Conjugation site modulates the in vivo stability and therapeutic activity of antibody-drug conjugates
Conjugation site modulates the in vivo stability and therapeutic activity of antibody-drug conjugates
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DOI：
10.1038/nbt.2108
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发表时间：
2012-02-01
影响因子：
46.9
通讯作者：
Junutula, Jagath R.
中科院分区：
文献类型：
--
作者：
Shen, Ben-Quan;Xu, Keyang;Junutula, Jagath R.
半胱氨酸中的反应性硫醇用于在抗体缀合物的产生中偶联马来酰亚胺接头。为了评估缀合位点的影响，我们在溶剂可及性和局部电荷不同的三个位点将半胱氨酸工程化到治疗性HER 2/neu抗体中。由于马来酰亚胺与白蛋白、游离半胱氨酸或谷胱甘肽中的反应性巯基交换，高度溶剂可及的位点迅速失去血浆中的缀合巯基反应性接头。相比之下，具有带正电荷的环境的部分可接近的位点促进了接头中琥珀酰亚胺环的水解，从而防止了这种交换反应。部分溶剂可及性和中性电荷的网站显示这两个属性。在小鼠乳腺肿瘤模型中，抗体偶联物的稳定性和治疗活性受到琥珀酰亚胺环水解的积极影响，并受到马来酰亚胺与血浆中巯基反应性成分交换的消极影响。因此，缀合位点的化学和结构动力学可以通过调节抗体-接头界面的稳定性来影响抗体缀合物性能。
The reactive thiol in cysteine is used for coupling maleimide linkers in the generation of antibody conjugates. To assess the impact of the conjugation site, we engineered cysteines into a therapeutic HER2/neu antibody at three sites differing in solvent accessibility and local charge. The highly solvent-accessible site rapidly lost conjugated thiol-reactive linkers in plasma owing to maleimide exchange with reactive thiols in albumin, free cysteine or glutathione. In contrast, a partially accessible site with a positively charged environment promoted hydrolysis of the succinimide ring in the linker, thereby preventing this exchange reaction. The site with partial solvent-accessibility and neutral charge displayed both properties. In a mouse mammary tumor model, the stability and therapeutic activity of the antibody conjugate were affected positively by succinimide ring hydrolysis and negatively by maleimide exchange with thiol-reactive constituents in plasma. Thus, the chemical and structural dynamics of the conjugation site can influence antibody conjugate performance by modulating the stability of the antibody-linker interface.