中文摘要：天门冬酰胺酶（ASNase）是一种治疗急性淋巴细胞白血病（ALL）的生物制剂。然而，它具有不良的副作用，如药效短，对蛋白酶敏感及免疫原性。本研究通过在基于p22的噬菌体类病毒颗粒(VLPs)中进行ASNase的基因定向包裹(ASNase- p22纳米反应器)来克服这种不足。ASNase-P22由58.4+/-7.9%的外壳蛋白和41.6+/-8.1%的四聚体ASNase组成。ASNase-P22的Km和Kcat值分别比游离酶高15倍和2倍。得到Kcat/Km值为2.19 × 10(5) M-1 s(-1)。37℃孵育12天，60%体积样本的ASNase-P22聚集；而相同条件下商品天冬酰胺酶则完全聚集。ASNase-P22在37℃下保持稳定达24小时，与是否存在人血清（HBS）无关，也与ASNase-P22纳米反应器是否未包被或聚乙二醇化无关。最后，本文发现ASNase-P22对白血病细胞系MOLT-4具有浓度依赖性的细胞毒性。本文首次将ASNase封装在VLPs中，用于对抗ALL病毒，前景很好。
外文摘要：Asparaginase (ASNase) is a biopharmaceutical for Acute Lymphoblastic Leukemia (ALL) treatment. However, it shows undesirable side effects such as short lifetimes, susceptibility to proteases, and immunogenicity. Here, ASNase encapsidation was genetically directed in bacteriophage P22-based virus-like particles (VLPs) (ASNase-P22 nanoreactors) as a strategy to overcome these challenges. ASNase-P22 was composed of 58.4 +/- 7.9% of coat protein and 41.6 +/- 8.1% of tetrameric ASNase. Km and Kcat values of ASNase-P22 were 15- and 2-fold higher than those obtained for the free enzyme, respectively. Resulting Kcat/Km value was 2.19 x 10(5) M-1 s(-1). ASNase-P22 showed an aggregation of 60% of the volume sample when incubated at 37 degrees C for 12 days. In comparison, commercial asparaginase was completely aggregated under the same conditions. ASNase-P22 was stable for up to 24 h at 37 degrees C, independent of the presence of human blood serum (HBS) or whether ASNase-P22 nanoreactors were uncoated or PEGylated. Finally, we found that ASNase-P22 caused cytotoxicity in the leukemic cell line MOLT-4 in a concentration dependent manner. To our knowledge, this is the first work where ASNase is encapsulated inside of VLPs, as a promising alternative to fight ALL.
外文关键词：asparaginase nanoreactors；ALL treatment；P22 virus-like particles
作者：Diaz-Barriga, C；Villanueva-Flores, F；Quester, K；Zarate-Romero, A；Cadena-Nava, RD；Huerta-Saquero, A
作者单位：Univ Nacl Autonoma Mexico
期刊名称：PHARMACEUTICS
期刊影响因子：4.421
出版年份：2021
出版刊次：5
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