Metabolic fate and oral dosing feasibility of the formyl peptide receptor 2 agonist WKYMVm: An integrated analytical and kinetic modeling approach
- Eur J Pharm Sci. 2026 Aug 1:223:107578. doi: 10.1016/j.ejps.2026.107578.
- 1. College of Pharmacy and Research Institute of Pharmaceutical Sciences, Seoul National University, Seoul, 08826, Republic of Korea.
- 2. Department of Biological Sciences, Sungkyunkwan University, Suwon, 16419, Republic of Korea.
- 3. College of Pharmacy and Research Institute of Pharmaceutical Sciences, Seoul National University, Seoul, 08826, Republic of Korea. Electronic address: [email protected].
Peptide-based drug candidates are often susceptible to enzymatic degradation in the gastrointestinal environment and biological matrices, while exhibiting limited intestinal permeability. In developing strategies to overcome these limitations, it is important to quantitatively evaluate the factors contributing to low systemic exposure and to understand the metabolic fate of peptides. This study utilized an integrated analytical approach combining quantitative analysis via liquid chromatography tandem-mass spectrometry (LC-MS/MS) and metabolite profiling via LC-quadrupole time-of-flight MS (LC-QTOF-MS) to characterize the metabolic fate and evaluate the oral dosing feasibility of WKYMVm (Wm), a formyl peptide receptor 2 agonist. A validated LC-MS/MS assay enabled quantification of Wm in mouse plasma with acceptable accuracy (95.2-115.7%) and precision (<8.6%). LC-QTOF-MS analysis identified eight metabolites (M1-M8) in mouse plasma, predominantly formed via sequential N-terminal cleavage with oxidative modifications. Plasma stability data were then simultaneously fitted to a structural parent-metabolite kinetic model, which identified the formation of M4 as the predominant pathway (fraction metabolized (fm) = 0.501), followed by M2 (fm = 0.262), indicating preferential cleavage at the Lys-Tyr bond. To predict oral dosing feasibility, Caco-2 bi-directional transport data were analyzed using a catenary model, while luminal degradation and hepatic metabolism were estimated using simulated gastric and intestinal fluids and mouse liver S9 fraction. When the experimentally obtained parameters were incorporated into simplified models, the results revealed limited intestinal and hepatic availability under the tested conditions. This integrated analytical and modeling approach identified the metabolic liabilities of Wm, informing the design of Wm analogs to enhance metabolic stability.