The study was designed to compare amylin hormone levels and some biochemical parameters in the serum of Iraqi patient with hypothyroidism (hypo) with and without Type 2 diabetes mellitus (T2DM). This study included ninety subjects and was divided into two patient groups: group I, which included 30 hypo without T2DM, and group II, which included 30 hypo with T2DM. Group III included 30 apparently healthy controls. The age range is 40-60 years. A significant increase in body mass index (BMI), fasting blood glucose (FBG), total cholesterol (TC), low-density lipoprotein (LDL), very low density lipoprotein (VLDL), triglyceride (TG), amylin hormone, and thyroid-stimulating hormone (TSH) in patients with hypo and hypo with T2DM (groups I and II) when compared to healthy controls (group III), and a significant decrease in the level of HDL in patient groups (I and II) when compared to healthy control groups. The amylin hormone receiver operating characteristic (ROC) curve showed a clear cut-off value (45.304 and 2) when calculated in hypo with and without T2DM compared with control groups, respectively. While the ROC curve showed a clear cut-off value (50.661) when compared in hypo with and without T2DM groups. Amylin is a hormone marker of glucose homeostasis, so it may be surrogate novel biomarker for all other traditional biomarkers for the prediction of diabetes and thyroid dysfunction.