Background: Rheumatoid Arthritis is a chronic joint synovial arthritic joint deterioration. The identification of novel biomarkers is crucial for improving diagnostic accuracy and understanding disease pathogenesis. Spexin, a recently discovered adipokine, has been implicated in metabolic and inflammatory processes, While Annexin A2 is recognized to have a pro-inflammatory function in the pathophysiology of RA, Spexin, a recently identified adipokine, has been linked to metabolic and inflammatory processes. Objective: The objective of this study was to investigate serum spexin levels and their correlation with Annexin A2 in males with rheumatoid arthritis compared to healthy controls, as well as to evaluate the diagnostic efficacy of these biomarker1s. Methods: A case-control study was undertaken including 42 male RA patients and 42 age- and BMI-matched healthy male controls (45-70 years). Serum concentrations of spexin, Annexin A2, anti-cyclic citrullinated peptide (ACCP), erythrocyte sedimentation rate (ESR), and uric acid were examined. Receiver operating characteristic (ROC) curve analysis, Pearson correlation, and independent t-tests were among the statistical methods used. Results: Results: The concentrations of serum spexin were significantly decreased in the RA group compared to the control group (4.201 ± 1.350 vs. 5.988 ± 1.666 ng/mL, p < 0.0001). On the other hand, the concentrations of serum Annexin A2 were significantly increased in the RA group (30.73 ± 6.543 vs. 21.21 ± 5.505 ng/mL, p < 0.001). Spexin had only poor analytical sensitivity (AUC = 0.221), whereas Annexin A2 had a good diagnostic value (AUC = 0.868). Conclusion: Lower serum levels of spexin and higher levels of Annexin A2 have been linked to RA in males. The inverse relationship between spexin and Annexin A2 levels implies possible interaction in the pathophysiology of RA. Annexin A2 can be used for diagnosing RA, whereas spexin may act as a treatment target.