(M1130-01-01) Development and Validation of a Sensitive LC-MS/MS Method for the Quantification of SGR-1505 in Human Plasma to Support Clinical Pharmacokinetic Studies

Purpose: SGR-1505 is a novel, selective MALT1 inhibitor under investigation for the treatment of B-cell malignancies, such as lymphoma and leukemia. To support clinical development, precise pharmacokinetic (PK) assessments are crucial for evaluating drug exposure and therapeutic efficacy. To this end, a sensitive and specific bioanalytical method was developed and validated for the quantification of SGR-1505 in human plasma. This abstract provides an overview of the validation of a liquid chromatography-tandem mass spectrometry (LC-MS/MS) method used for the quantitative determination of SGR-1505. The method was validated according to current FDA and ICH guidelines to ensure it met the regulatory requirements for bioanalytical testing in drug development. Parameters evaluated included selectivity, sensitivity (low limit of quantification or LLOQ), linearity, accuracy, precision (intra- and inter-day), recovery, and stability under different storage and handling conditions, such as freeze/thaw cycles, short-term and long-term storage, and in-matrix stability. The validation process was designed to ensure method robustness under conditions in which clinical samples are handled. The validated method demonstrated acceptable performance across all parameters. The LLOQ of 10.0 ng/mL was achieved, allowing for the detection and quantification of low concentrations of SGR-1505 in plasma. Linearity was confirmed over a dynamic range from 10.0 to 10,000 ng/mL, ensuring accurate quantification across a broad concentration range. Accuracy and precision were well within the predefined acceptance criteria for both in intra- and inter-day assessments, confirming the method's reproducibility. Extraction recovery assessments exhibited variability across the concentration range, with observed recoveries spanning from approximately 123.5% to 163.9%. The elevated recovery percentages, particularly pronounced at lower concentrations, suggest a potential matrix effect during the extraction process. Despite these findings, overall method performance remained acceptable, as demonstrated by passing precision and accuracy across the dynamic range and satisfactory results in other validation tests. The stability of SGR-1505 was demonstrated for stock solutions and in plasma under various conditions including benchtop, multiple freeze/thaw cycles, as well as short- and long-term frozen storage, ensuring reliable results during sample collection, processing and storage. Incurred Sample Reanalysis was also evaluated during clinical sample analysis verifying the validated methods accuracy and reproducibility. This validated LC-MS/MS method has been successfully implemented in clinical studies of SGR-1505. This method is now routinely used to analyze plasma samples from ongoing studies, providing essential pharmacokinetic data for assessing drug exposure and informing dosage regimens.

Methods: A bioanalytical method using liquid chromatography-tandem mass spectrometry (LC-MS/MS) was developed to quantify SGR-1505 in human plasma. Sample preparation involved protein precipitation, followed by centrifugation. The supernatant was analyzed by LC-MS/MS using a reversed-phase column and gradient elution with appropriate mobile phases. Detection was performed in positive ionization mode using multiple reaction monitoring (MRM) transitions specific to SGR-1505 and an internal standard. Calibration standards ranged from 10.0 to 10,000 ng/mL. Quality control (QC) samples were prepared at four levels: LLOQ, low, mid, and high. The method was validated according to FDA and ICH guidelines to support clinical studies. Validation parameters included selectivity, sensitivity, linearity, accuracy, precision (intra- and inter-day), recovery, matrix effects, and stability under various conditions (benchtop, freeze/thaw, short-term and long-term frozen storage, and stock solution stability). Incurred Sample Reanalysis (ISR) was also performed to confirm method reproducibility during clinical sample analysis. All validation activities adhered to Good Laboratory Practice (GLP). The method demonstrated consistent performance across all parameters and is now routinely used to support pharmacokinetic assessments in clinical trials of SGR-1505.

Results: The validated LC-MS/MS method achieved an LLOQ of 10.0 ng/mL and demonstrated linearity across a range of 10.0 to 10,000 ng/mL (R² ≥ 0.99). Intra- and inter-day accuracy and precision met acceptance criteria, confirming method reproducibility. Mean extraction recoveries ranged from ~123.5% to 163.9%, with higher recovery at lower concentrations, indicating potential matrix effects; however, overall assay performance remained acceptable. SGR-1505 was stable under various conditions, including benchtop, three freeze/thaw cycles, and short- and long-term frozen storage. Stock solution stability was also confirmed. Incurred Sample Reanalysis (ISR) met predefined criteria, supporting the method’s reliability during clinical sample analysis. The validated method consistently met regulatory standards and is currently used in clinical trials to generate pharmacokinetic data for SGR-1505.

Conclusion: A robust, sensitive, and specific LC-MS/MS method was successfully developed and validated for the quantification of SGR-1505 in human plasma, in accordance with FDA and ICH guidelines. The method demonstrated acceptable performance across all validation parameters, including accuracy, precision, linearity, stability, and sensitivity, with an LLOQ of 10.0 ng/mL and a dynamic range of 10.0–10,000 ng/mL. Despite elevated recovery values indicating potential matrix effects, overall method integrity and reliability were maintained. The validated method has been effectively implemented in clinical studies and is being routinely used to generate critical pharmacokinetic data to support the clinical development of SGR-1505 in the treatment of B-cell malignancies.

References: Validation of STC-7505 in Human Plasma