接受用于恶性肿瘤的高剂量甲氨蝶呤（HDMTX）的患者面临各种并发症，包括肾毒性、肝毒性、口腔炎、骨髓毒性、神经症状和死亡。Glucarpidase是重组羧肽酶G2（CPG2），可将MTX转化为无毒代谢产物。本研究调查了载体类型、基因优化、方向和宿主对CPG2表达的作用。对包含glucarpidase的各种治疗方案的有效性进行了分类，并提供了基于精准医疗的剂量调整的观点。强调了将CPG2与细胞穿透肽、人血清白蛋白和聚合物如PEG和dextran结合以提高递送效果、稳定性和生产更好的CPG2变体。传达了将CPG2与针对肿瘤特异性抗原的F(ab՜)2或scFv抗体片段结合，以及使用抗体导向的酶前药物疗法（ADEPT）实现肿瘤靶向药物递送的相应前药的信息。报道了通过添加对肿瘤过度表达的蛋白酶敏感的前域、抗CPG2抗体、具有免疫系统不可识别表位的CPG2突变体和保护性聚合物来减少非靶点效应和重复ADEPT周期的尝试。描述了基因导向的酶前药物疗法（GDEPT）中的细胞内cpg2基因表达以及GDEPT载体安全性和转染效果的问题。引入了一种新型的双功能平台，利用工程化的CAR-T细胞微药库，称为合成酶武装杀手（SEAKER）细胞，表达CPG2以在肿瘤微环境中激活前药。综合本综述中的整合数据和在新型药物递送系统中应用组合策略，定义了ADEPT、GDEPT和SEAKER细胞疗法的未来发展方向以及其中CPG2的应用。版权所有 © 2023 作者。Elsevier Masson SAS出版。保留所有权利。

Patients receiving high-dose methotrexate (HDMTX) for malignancies are exposed to diverse complications, including nephrotoxicity, hepatotoxicity, mucositis, myelotoxicity, neurological symptoms, and death. Glucarpidase is a recombinant carboxypeptidase G2 (CPG2) that converts MTX into nontoxic metabolites. In this study, the role of vector type, gene optimization, orientation, and host on the expression of CPG2 is investigated. The effectiveness of various therapeutic regimens containing glucarpidase is classified and perspectives on the dose adjustment based on precision medicine are provided. Conjugation with cell-penetrating peptides, human serum albumin, and polymers such as PEG and dextran for delivery, higher stability, and production of the biobetter variants of CPG2 is highlighted. Conjugation of CPG2 to F(ab՜)2 or scFv antibody fragments against tumor-specific antigens and the corresponding prodrugs for tumor-targeted drug delivery using the antibody-directed enzyme prodrug therapy (ADEPT) is communicated. Trials to reduce the off-target effects and the possibility of repeated ADEPT cycles by adding pro-domains sensitive to tumor-overexpressed proteases, antiCPG2 antibodies, CPG2 mutants with immune-system-unrecognizable epitopes, and protective polymers are reported. Intracellular cpg2 gene expression by gene-directed enzyme prodrug therapy (GDEPT) and the concerns regarding the safety and transfection efficacy of the GDEPT vectors are described. A novel bifunctional platform using engineered CAR-T cell micropharmacies, known as Synthetic Enzyme-Armed KillER (SEAKER) cells, expressing CPG2 to activate prodrugs at the tumor niche is introduced. Taken together, integrated data in this review and recruiting combinatorial strategies in novel drug delivery systems define the future directions of ADEPT, GDEPT, and SEAKER cell therapy and the placement of CPG2 therein.Copyright © 2023 The Authors. Published by Elsevier Masson SAS.. All rights reserved.