A1480LS is a peripherally restricted, orally active covalent and irreversible inhibitor of both DAGLα and DAGLβ, exhibiting IC50 values of 6 nM and 4 nM at human targets, respectively, and IC50 values ≤15 nM across mice, rats, dogs, monkeys, and human systems. A1480LS reduces levels of 2-arachidonoylglycerol, arachidonic acid, and eicosanoids derived from cyclooxygenase and lipoxygenase pathways. It inhibits the injury-induced production of 2-arachidonoylglycerol and arachidonic acid in peripheral sciatic nerves and reduces the response of high-threshold and wide dynamic range-like dorsal horn neurons to mechanical stimuli. Moreover, A1480LS alleviates pain behaviors in rat models of inflammatory pain, neuropathic pain, and chemotherapy-induced peripheral neuropathy.