► Deltorphins analogs (DEL-6 and DK-4) induced antinociception in thermal pain. ► These effects were attenuated by MOR and DOR antagonists. ► Non-effective doses of DEL-6 and morphine produced additive antinociceptive action. ► Both deltorphins analogs produced low cross-tolerance with morphine. ► Deltorphins analogs involve MOR and DOR to exert their antinociceptive effects. The antinociceptive effects of analogs of deltorphins: cyclo(Nδ,Nδ-carbonyl-d-Orn2, Orn4)deltorphin (DEL-6) and deltorphin II N-(ureidoethyl)amide (DK-4) after intracerebroventricular (i.c.v.) administration were investigated in the tail-immersion test in rats. Morphine, the most commonly used μ-opioid receptors (MOR) agonist, was employed as a reference compound. The contribution of the MOR, δ-(DOR) and κ-opioid receptors (KOR) in antinociceptive effects of the deltorphins analogs was studies using selective antagonists of these receptors. The results indicated that DK-4 (5, 10 and 20nmol) and DEL-6 (5, 10 and 20nmol) were the most effective in alleviating thermal pain at the dose of 20nmol. The antinociceptive potency of DEL-6 at the dose of 20nmol was approximately equal but DK-4 at the dose of 20nmol was less effective than morphine at the dose of 13nmol. DOR antagonist – naltrindole (NTI, 5nmol) very strongly and, to the lower extent MOR antagonist – β-funaltrexamine (β-FNA, 5nmol), inhibited antinociceptive effect of DK-4 (20nmol). In turn, β-FNA was more potent than NTI in inhibition of the antinociceptive effects of DEL-6. Co-administration of DEL-6 and morphine at doses of 5nmol, which do not produce measurable antinociception, generated additive antinociceptive effect. Chronic intraperitoneal (i.p.) injection of morphine (9 days) displayed a marked analgesic tolerance to the challenge dose of morphine and a slight cross-tolerance to challenge doses of DEL-6 and DK-4, given i.c.v. These findings indicate that the new deltorphin analogs recruit DOR and MOR to attenuate the nociceptive response to acute thermal stimuli.