Hecogenin acetate (HA), a steroidal acetylated-sapogenin, has an analgesic profile already assigned, but its low water solubility and short half-life limit its use in chronic conditions. beta-cyclodextrin (beta-CD) can improve the chemical and pharmacological property of nonpolar compounds such as HA. Therefore, a HA complexed with beta-CD (HA-CD) was prepared and characterized by thermal, morphological and spectroscopic analysis. A model of chronic musculoskeletal pain was induced by means of two injections of pH 4.0 saline (20 mu L) into the left gastrocnemius 5 days apart was used. After confirming hyperalgesia, male mice were treated with HA, HA-CD (20 mg/kg; p.o.) or vehicle (saline 0.9%, p.o.). Motor coordination tests, substance P (SP) levels in a lumbosacral (L4-S2) spinal cord sample and a docking study were equally assessed to check for a possible action on the opioid receptors. Oral pretreatment with HA or HA-CD produced a significant antinociceptive (p < 0.01) profile and also decreased mechanical hyperalgesia, with HA-beta-CD showing significantly better effects when compared to HA alone (p < 0.05). The interaction between HA and the opioid receptor (MU, Kappa, Delta)was corroborated by the docking study. Moreover, the SP level was reduced in a spinal cord sample. Our findings suggest that beta-CD can improve the anti-hyperalgesic effect of HA in an animal model of musculoskeletal pain. (C) 2016 Elsevier Ltd. All rights reserved.