Background: L-glutamine (Gln) suppresses inflammation via rapid up-regulation of MAPK phosphatase (MKP)-1, deactivating p38 and JNK mitogen-activated protein kinases (MAPKs). However, the high dosage required for this may cause serious side effects. Objective: To facilitate reduced Gln intake, we developed a less-hydrolysable Gln derivative, α, δ-N-acetyl-glutamine (α, δ-NAG), which is resistant to the hydrolytic action of glutaminase. Methods: We developed α, δ-NAG by substituting the NH ₂ group in α-chain and δ-amide group of Gln with acetyl groups. We employed the ovalbumin model, previously developed by us, to examine sequential asthmatic events, including neutrophilia/Th1 and eosinophilia/Th2 responses. MKP-1 was knocked down using small-interfering RNA (siRNA). Gln levels and intracellular calcium concentration ([Ca ²⁺] _i) were analysed using multiple reaction monitoring chromatograms and confocal laser scanning microscopy, respectively. Results: Oral administration of α, δ-NAG and Gln suppressed all the parameters at 0.2 and 2 g/kg body weight, respectively. MKP-1 siRNA abrogated the beneficial effects of α, δ-NAG. α, δ-NAG up-regulated MKP-1 in an ERK MAPK-dependent manner. α, δ-NAG transiently increased [Ca ²⁺] _I, resulting in increased Ras activity. Inhibition of Gα _q, a G-protein subfamily, abrogated the effects of α, δ-NAG on [Ca ²⁺] _I and Ras activity. Inhibition of Gα _q, Ca ²⁺, and Ras abrogated the effects of α, δ-NAG, such as signalling pathways (ERK phosphorylation and MKP-1 up-regulation) and clinical signs (neutrophilia/Th1 responses) in asthmatic mice. Conclusion: α, δ-NAG exhibits strong anti-inflammatory activity (~ 10,000-fold stronger than that of Gln), likely attributable to its up-regulation of MKP-1 by activating pathways involving the G protein-coupled receptor (GPCR)/Gα _q/Ca ²⁺/Ras/ERK cascade.