To exert analgesic action, paracetamol requires complex metabolization to produce a brain specific lipoaminoacid compound, AM404, which targets central TRPV1 receptors. Lipoaminoacids are also known to induce analgesia through T-type channel inhibition (Cav3.2). In this study we show that the antinociceptive effect of paracetamol is lost in mice when supraspinal Cav3.2 channels are inhibited. Therefore, we hypothesized a relationship between supraspinal Cav3.2 and TRPV1, via AM404, which mediates the analgesic effect of paracetamol. AM404 is able to activate TRPV1 and weakly inhibits Cav3.2. Interestingly, activation of TRPV1 induces a strong inhibition of Cav3.2 current. Supporting this, intracerebroventricular administration of AM404 or capsaicin produces antinociception that is lost in Cav3.2(-/-) mice. Our study, for the first time, (1) provides a molecular mechanism for the supraspinal antinociceptive effect of paracetamol, (2) identifies the relationship between TRPV1 and Cav3.2 and (3) discloses supraspinal Cav3.2 inhibition as a potential pharmacological strategy to alleviate pain.