T-type Ca²⁺ channels (T-channels) serve as targets for treatment of pain. We have reported that 6-prenylnaringenin (6-PNG), a hop component, and its derivative, KTt45, inhibit T-channels. Interestingly, many of cannabinoids block T-channels, while some of T-channel blockers stimulate cannabinoid CB₁ or CB₂ receptors. Thus, we compared the effects of 6-PNG and KTt45 as well as cannabinoids on Ca_v3.2 T-channels and CB₁/CB₂ receptors. In Ca_v3.2-expressing HEK293 cells, 6-PNG, KTt45, ACEA, a CB₁ agonist, and cannabidiol, a possible CB₁ antagonist/CB₂ inverse agonist, blocked T-currents, as assessed by whole-cell recordings; IC₅₀ values (μM) were 0.69, 0.41, 0.57 and 0.64, respectively. Neither 6-PNG nor KTt45, exhibited agonistic activity in CB₁-expressing HEK293 cells, whereas 6-PNG at 0.3-1 μM, but not KTt45, agonistic activity in CB₂-expressing CHO cells. In distinct mouse models for neuropathic and bladder pain, i.p. administration of 6-PNG at 20-30 mg/kg and KTt45 at 10-30 mg/kg exhibited analgesic/anti-allodynic activity. Thus, 6-PNG and KTt45, T-channel blockers, are useful for treatment of intractable pain, and 6-PNG is considered a mixed T-channel blocker/CB₂ agonist.