Blackman Lewis D, Varlas Spyridon, Arno Maria C, Houston Zachary H, Fletcher Nicholas L, Thurecht Kristofer J, Hasan Muhammad, Gibson Matthew I, O'Reilly Rachel K
Department of Chemistry, University of Warwick, Gibbet Hill Road, CV4 7AL Coventry, United Kingdom.
School of Chemistry, University of Birmingham, Edgbaston, Birmingham B15 2TT, United Kingdom.
ACS Cent Sci. 2018 Jun 27;4(6):718-723. doi: 10.1021/acscentsci.8b00168. Epub 2018 May 16.
Covalent PEGylation of biologics has been widely employed to reduce immunogenicity, while improving stability and half-life . This approach requires covalent protein modification, creating a new entity. An alternative approach is stabilization by encapsulation into polymersomes; however this typically requires multiple steps, and the segregation requires the vesicles to be permeable to retain function. Herein, we demonstrate the one-pot synthesis of therapeutic enzyme-loaded vesicles with size-selective permeability using polymerization-induced self-assembly (PISA) enabling the encapsulated enzyme to function from within a confined domain. This strategy increased the proteolytic stability and reduced antibody recognition compared to the free protein or a PEGylated conjugate, thereby reducing potential dose frequency and the risk of immune response. Finally, the efficacy of encapsulated l-asparaginase (clinically used for leukemia treatment) against a cancer line was demonstrated, and its biodistribution and circulation behavior was compared to the free enzyme, highlighting this methodology as an attractive alternative to the covalent PEGylation of enzymes.
生物制品的共价聚乙二醇化已被广泛用于降低免疫原性，同时提高稳定性和半衰期。这种方法需要共价蛋白质修饰，从而产生一个新的实体。另一种方法是通过封装到聚合物囊泡中来实现稳定化；然而，这通常需要多个步骤，并且这种分离要求囊泡具有渗透性以保持功能。在此，我们展示了使用聚合诱导自组装（PISA）一锅法合成具有尺寸选择性渗透性的负载治疗性酶的囊泡，使封装在其中的酶能够在受限区域内发挥作用。与游离蛋白质或聚乙二醇化缀合物相比，该策略提高了蛋白水解稳定性并降低了抗体识别，从而降低了潜在的给药频率和免疫反应风险。最后，证明了封装的L-天冬酰胺酶（临床用于白血病治疗）对癌细胞系的疗效，并将其生物分布和循环行为与游离酶进行了比较，突出了该方法作为酶共价聚乙二醇化的一种有吸引力的替代方法。
