(T1330-03-16) Implementing the Latest Available Technologies in High Throughput through In Vitro ADME DMPK

Purpose: The enormous surge in the quantity of new chemical entities requested in the early stages of drug discovery has created an urgent need for improved throughput. Reduction in acquisition times across all instrumentation for sample analysis can significantly accelerate workflows in the biotech and pharmaceutical industries. Specifically, in vitro absorption, distribution, metabolism, and elimination (ADME) assays are routinely employed from hit identification through lead optimization to minimize attrition of potential drug candidates during development. Since the 1990s, LC/MS has been extensively used for the quantitation of small molecules in both in vitro ADME and bioanalytical PK groups. Recent advances in triple quadrupole mass spectrometers—such as improved sensitivity, faster scan speeds, and expanded dynamic range—have been complemented by innovations in front-end technologies, including multiplexed LC systems, RapidFire™, ADDA, LS1/2, and Echo-MS. In this work we have shown the progression in LC/MS technologies and its successful implementation within the in vitro ADME group at Biogen resulting in substantial improvements in the turnaround times of ADME data. Strategies for streamlining ADME assessment of compounds in discovery and pre-clinical development studies are presented within.

Methods: A comparative evaluation was conducted between the newer LSII (Sound Analytics) and the older ADDA system (Apricot Designs). A trap and elute chromatography technique was employed to improve sensitivity via increased peak intensities in liquid chromatography (LC) systems.  Both front-ends were connected to separate API-5500 (Sciex) mass spectrometer. The assay of choice was microsomal stability using rat and human liver microsomes.

Results: Following method optimization, the LSII method increased the throughput by 38%, reducing per-sample analysis time by 9 seconds. The total run time to analyze a 96 well plate decreased from 29 to 18 minutes.

Conclusion: Over 97% of the 204 samples analyzed of the microsomal half-life values were within 2-fold differences between the two methods. Attempts to implement an even faster method (4-4-7) resulted in peak loss due to insufficient column equilibration, highlighting the need to balance speed with data quality.

Trap and Elute Method 9915 (ADDA): Equilibrate time: 9 sec, desalt time: 9 sec, sweep time: 15 sec


Trap and Elute Method 558 (LS-II): Equilibrate time: 5 sec, desalt time: 5 sec, sweep time: 8 sec

Microsomal Stability Half-life Values Comparison in Rat and Human from the LSII vs. ADDA methods