Abstract

Catalase is an antioxidant enzyme with potential application in photoprotection, skin aging, and skin disorders as vitiligo. However, as a large and hydrophilic molecule, catalase topical application is hampered by poor skin penetration. In this sense, nanostructured drug delivery systems such as polymersomes (polymeric vesicles) can significantly promote the enzyme access to skin. Since the concept of designing quality in early stages of biotechnological products development is a regulatory demand over the world, in this study, we employed Quality by Design (QbD) principles to design catalase-loaded polymersomes for topical products. QbD and Design of Experiments (DoE) approaches were used to investigate the impact of stirring speed (SS =750 and 1500 rpm), stirring time (ST = 24 and 48 h), and concentration of catalase (CAT = 0.48 and 0.24 mg/mL) in pharmaceutical development critical quality attributes of PEO₅PPO₆₈PEO₅ polymersomes for topical application. The design space counterplot of hydrodynamic diameter (200 ≤ HD ≤ 400), encapsulation efficiency (2≤ %EE ≤ 5), polydispersity index (0.1 ≤ PDI ≤ 0.3), as a function of ST and CAT indicated that polymersomes with optimized characteristics could be developed with 0.24 ≥ CAT ≤ 0.40; 40 ≥ ST ≤ 48 and SS= 800 rpm. Encapsulation efficiency (%EE) was determined directly by size exclusion chromatography purification of the polymersomes, and the highest value was %EE = 4.72 ± 0.07 %, nonetheless catalase as an enzyme is effective at very low concentrations. The results point out time and cost-saving strategies that could serve as an initial step to develop a robust industrial process for the production of catalase-loaded topical polymersomes.

摘要

过氧化氢酶是一种抗氧化酶，在光保护、皮肤老化和白斑等皮肤疾病方面具有潜在的应用价值。然而，作为一种大分子和亲水性分子，过氧化氢酶局部应用受到皮肤渗透性差的阻碍。从这个意义上说，纳米结构的药物递送系统如聚合物囊泡（聚合物囊泡）可以显着促进酶进入皮肤。由于在生物技术产品开发的早期阶段设计质量的概念是全世界的监管要求，因此在本研究中，我们采用质量源于设计 (QbD) 原则来设计用于外用产品的过氧化氢酶负载的聚合物囊泡。QbD 和实验设计 (DoE) 方法用于研究搅拌速度（SS = 750 和 1500 rpm）、搅拌时间（ST = 24 和 48 小时）和过氧化氢酶浓度（CAT = 0.48 和 0.₅ PPO ₆₈ PEO ₅用于局部应用的聚合物囊泡。流体动力学直径 (200 ≤ HD ≤ 400)、包封效率 (2≤ %EE ≤ 5)、多分散指数 (0.1 ≤ PDI ≤ 0.3) 作为 ST 和 CAT 的函数的设计空间反图表明具有优化特性的聚合物囊泡可以发展为 0.24 ≥ CAT ≤ 0.40；40 ≥ ST ≤ 48 且 SS= 800 rpm。封装效率 (%EE) 由聚合物囊泡的尺寸排阻色谱纯化直接确定，最高值为 %EE = 4.72 ± 0.07 %，尽管如此，过氧化氢酶作为一种酶在非常低的浓度下也是有效的。结果指出了节省时间和成本的策略，这些策略可以作为开发用于生产负载过氧化氢酶的局部聚合物囊泡的稳健工业过程的第一步。