Δ12-Prostaglandin J3 (Δ12-PGJ3), a cyclooxygenase-2 (COX-2) metabolite of omega-3 polyunsaturated fatty acid (PUFA), eicosapentaenoic acid (EPA; 20:5), has shown promising anti-leukemic properties in mice and leukemia patient samples with regard to its ability to selectively ablate leukemia stem cells (LSCs). With procedures already established to help chemically synthesize Δ12-PGJ3 from PGD3, this study focused on the novel enzymatic synthesis of Δ12-PGJ3 that would be essential for its molecular characterization and comparison with the chemically synthesized compound.
Although initial reactions of Δ12-PGJ2 were unsuccessful, appropriate solvent systems were formulated for separation of lipids from enzymatically-synthesized products. Using recombinant human COX-2 expressed in a baculovirus system and bacterially expressed human recombinant hematopoietic PGD synthase (H-PGDS) enzymes, Δ12-PGJ3 was successfully synthesized from EPA. Enzymatically synthesized PGJ3 was confirmed by UV-spectroscopy, thin layer chromatography (TLC), and liquid chromatography-mass spectrometry (LC-MS/MS). This product will enable future studies on LSCs and will allow comparisons between enzymatic and synthetic Δ12-PGJ3.
