血浆中氟达拉滨定量存在显著基质效应：对 CAR-T 细胞治疗监测的启示

Event-free survival (EFS) in patients with B-cell acute lymphoblastic leukemia (B-ALL) significantly improves with adequate exposure (as Area Under the Curve: AUC 0-inf or AUC) to fludarabine (FLU) during lymphodepletion prior to chimeric antigen receptor (CAR) T-cell infusion. A cumulative AUC of FLU exceeding 14 mgxh/L is considered optimal, making therapeutic drug monitoring (TDM) crucial for personalized dosing and prolonged CAR T-cell persistence. RESEARCH DESIGN AND METHODS: We developed and validated an European Medicines Agency (EMA)-compliant high-performance liquid chromatography-tandem mass spectrometry method (HPLC-MS/MS) for FLU TDM, based on similar and published methods, to support clinicians during the patient conditioning phase. All EMA acceptance criteria were met.

Analysis of real plasma samples revealed unexpectedly high FLU concentrations and, consequently, an excessively high AUC. Analysis of plasma samples from a child receiving FLU therapy highlighted the necessity for accurate drug response verification in real matrices. Preparation of quality controls in FLU-free plasma consistently failed without preliminary dilution. Analysis of internal standard range variation enabled more precise and accurate analyses.

This experience highlights the importance of complementing standard validation tests with analyses in each patient's plasma, collected immediately before FLU infusion, to ensure reliable TDM to support CAR T-cell therapy. The success of a special treatment called CAR-T for people with leukemia depends on a few things. One of them is the amount of a drug called Fludarabine they received a few days beforehand. Patients with leukemia need several doses of a drug called Fludarabine. The right amount to give them depends on how much of the drug is already in their blood. To find this out, doctors must take several blood samples, which are then analyzed using special and often complicated lab methods. Getting these methods to work correctly requires skilled people and special equipment. Sometimes, however, hard-to-solve problems come up. These problems must be fixed very quickly and with only a small amount of blood to work with. This article shares a story about how some researchers did their best to handle one of these difficult situations. They believe they found a good solution, even with all the challenges and limited resources they faced.