Analgesic-response variability in chronic noncancer pain (CNCP) has been reported due to several biological and environmental factors. This study was undertaken to explore sex differences linked toOPRM1andCOMTDNA methylation changes and genetic variants in analgesic response. A retrospective study with 250 real-world CNCP outpatients was performed in which data from demographic, clinical, and pharmacological variables were collected. DNA methylation levels (CpG island) were evaluated by pyrosequencing, and their interaction with theOPRM1(A118G) andCOMT(G472A) gene polymorphisms was studied. A priori-planned statistical analyses were conducted to compare responses between females and males. Sex-differentialOPRM1DNA methylation was observed to be linked to lower opioid use disorder (OUD) cases for females (p= 0.006). Patients with lowerOPRM1DNA methylation and the presence of the mutant G-allele reduced opioid dose requirements (p= 0.001), equal for both sexes. Moreover,COMTDNA methylation levels were negatively related to pain relief (p= 0.020), quality of life (p= 0.046), and some adverse events (probability > 90%) such as constipation, insomnia, or nervousness. Females were, significantly, 5 years older with high anxiety levels and a different side-effects distribution than males. The analyses demonstrated significant differences between females and males related toOPRM1signalling efficiency and OUD, with a genetic–epigenetic interaction in opioid requirements. These findings support the importance of sex as a biological variable to be factored into chronic pain-management studies.