The proper management of pain is a critical issue in the practice of medicine. Despite the availability of a large number of analgesic medications, management of pain that is refractory to conventional treatments remains a challenge for both clinicians and surgeons. Botulinum neurotoxin (BoNT) has recently emerged as a potential novel approach to control pain. Animal studies have revealed a number of mechanisms by which BoNTs can influence and alleviate chronic pain, including inhibition of pain peptide release from nerve terminals and sensory ganglia, anti-inflammatory and antiglutaminergic effects, reduction of sympathetic neural discharge, and inhibition of muscle spindle discharge. In humans, prospective, placebo-controlled, double-blind studies have also provided evidence for effectiveness of BoNT therapy in a number of painful disorders. These include cervical dystonia, pelvic pain, low back pain, plantar fasciitis, postsurgical painful spasms, myofascial pain syndromes, migraine, and chronic daily headaches. Long-term studies on cervical dystonia and low back pain have demonstrated safety and sustained efficacy after repeated injections. This Review focuses on the analgesic effects of BoNT and the mechanisms of its pain control as revealed by animal models, and provides evidence-based data on the efficacy of BoNT therapy in various pain syndromes in humans.