Department of Biotechnology, National Institute of Pharmaceutical Education and Research (NIPER), Sector 67, S.A.S. Nagar, Mohali, 160062, Punjab, India.
Laboratory of Epigenetics and Diseases, Department of Pharmacology and Toxicology, National Institute of Pharmaceutical Education and Research (NIPER), Sector 67, S.A.S. Nagar, Mohali, 160062, Punjab, India.
Invest New Drugs. 2023 Oct;41(5):652-663. doi: 10.1007/s10637-023-01387-y. Epub 2023 Aug 3.
Recombinant human arginase I (rhArg I) have emerged as a potential candidate for the treatment of varied pathophysiological conditions ranging from arginine-auxotrophic cancer, inflammatory conditions and microbial infection. However, rhArg I have a low circulatory half-life, leading to poor pharmacokinetic and pharmacodynamic properties, which necessitating the rapid development of modifications to circumvent these limitations. To address this, polyethylene glycol (PEG)ylated-rhArg I variants are being developed by pharmaceutical companies. However, because of the limitations associated with the clinical use of PEGylated proteins, there is a dire need in the art to develop rhArg I variant(s) which is safe (devoid of limitations of PEGylated counterpart) and possess increased circulatory half-life. In this study, we described the generation and characterization of a fused human arginase I variant (FHA-3) having improved circulatory half-life. FHA-3 protein was engineered by fusing rhArg I with a half-life extension partner (domain of human serum albumin) via a peptide linker and was produced using P. pastoris expression system. This purified biopharmaceutical (FHA-3) exhibits (i) increased arginine-hydrolyzing activity in buffer, (ii) cofactor - independency, (iii) increased circulatory half-life (t) and (iv) potent anti-cancer activity against human cancer cell lines under in vitro and in vivo conditions.
重组人精氨酸酶 I（rhArg I）已成为治疗从精氨酸营养缺陷型癌症、炎症和微生物感染等多种病理生理状况的潜在候选药物。然而，rhArg I 的循环半衰期较短，导致其药代动力学和药效学性质较差，因此需要迅速开发修饰方法来克服这些限制。为了解决这个问题，制药公司正在开发聚乙二醇（PEG）化-rhArg I 变体。然而，由于与 PEG 化蛋白的临床应用相关的限制，迫切需要开发安全的 rhArg I 变体（没有 PEG 化对应物的限制），并具有延长的循环半衰期。在这项研究中，我们描述了一种融合的人精氨酸酶 I 变体（FHA-3）的产生和特性，该变体具有延长的循环半衰期。通过肽接头将 rhArg I 与半衰期延长伴侣（人血清白蛋白的结构域）融合，从而工程化 FHA-3 蛋白，并使用巴斯德毕赤酵母表达系统进行生产。这种纯化的生物制药（FHA-3）表现出：（i）在缓冲液中增加了精氨酸水解活性，（ii）辅酶非依赖性，（iii）延长了循环半衰期（t），以及（iv）在体外和体内条件下对人癌细胞系具有强大的抗癌活性。
