Background: Chronic pain is an urgent medical problem worldwide, resulting in long-term sick leave, low quality of life, and high socioeconomic costs. Despite the physiological, emotional, and financial burden of chronic pain, there is still a lack of efficient treatments. In this context, toxins isolated from arthropods are considered powerful tools because they have targets that are compatible with the impulse transmission of pain and can provide an interesting alternative in the development of more efficient analgesic treatments. Wasps are arthropods with potent venom that contains a complex mixture of compounds, including biogenic amines, proteins, and peptides. For decades, chemical identification and biological characterization of wasp venom peptides have accumulated considerable attention from researchers. Objective: The objective of this work was to evaluate the antinociceptive activity of the peptide fraction (Fpep) from low-molecular-weight compounds (LMWCs) from the venom of the Brazilian social wasp Pseudopolybia vespiceps testacea. Materials and Methods: P. vespiceps females were collected, and after euthanasia by freezing (−20°C), a total of 148 venom sacs were used to acquire only compounds with LMWCs. The antinociceptive effect of the Fpep from LMWCs was observed by two thermal tests: tail flick and hot plate measured in male Swiss mice. Results: Fpep showed dose-dependent antinociceptive activity in mice submitted to intracerebroventricular injection in two different models. Conclusion: These results revealed the significant potential impact of the LMWCs of the venom of Pseudopolybia vespiceps testacea, including neuroactive peptides that can be used as pharmacological resources for antinociceptive drug research.