Traumatic brain injury (TBI) is an acute brain lesion representing one of the main causes of mortality and disability worldwide. Searching for therapeutic strategies that shrink the inflammatory response after TBI is key to prevent and/or ameliorate secondary brain injury and to improve patients outcome. After TBI, there is a fast activation of innate immunity, where microglia responds to damage-associated molecular patterns such as ATP by P2X7 purinergic receptors. The P2X7-NLRP3 inflammasome axis has been identified as one of the main actors in neuroinflammation. Thus, the aim of this study was to validate P2X7 as a potential therapeutic target in TBI, as well as to evaluate new non-nucleotide purine derivative compounds as P2X7 antagonists in models of TBI both in vitro and in vivo. p2x7-/- mice subjected to the closed head injury model showed worse brain transcriptomic proinflammatory profile as well as poorer neuroconductual score 24h after TBI in comparison to WT mice. However, the animals treated with the potent P2X7 antagonist JNJ-47965567 (30mg/kg) 30min after TBI, presented improved neurological and inflammatory parameters. These apparently differing results depicted the relevance of P2X7 in a time-dependent control of traumatic lesion, further evaluated in a second pharmacological approach. A series of purine derivative compounds, divided into etanoil and sulfonil families, were evaluated as P2X7 antagonists. ITH15004 and ITH15003 were the most effective compounds preventing IL-1β release from primary mixed glial cultures and were tested in vivo. Altogether, these results highlight P2X7 implication as a potential therapeutic target to modulate neuroinflammation in TBI.