In this study, non-enzymatic glucose sensor composed of two-dimensional (2D) borophene-decorated polypyrrole (PPy) nanocomposites (NCs) was developed. The prepared PPy-borophene NCs were prepared using a low-cost sonication method. The sensing performance of the PPy-borophene NCs was investigated by the cyclic voltammetry (CV) technique against various biomolecules such as glucose, maltose, lactose, fructose, and urea. According to the electrochemical results, it was observed that in the glucose concentration range of 1.5 to 24 mM within a voltammetric cycle of 1 min, the PPy-based sensor and PPy-borophene NCs-based sensor exhibited sensitivities of 11.88 μAmM−1 cm−2 and 213.42 μAmM−1 cm−2, respectively. The detection limits of the PPy-based and PPy-borophene NCs-based sensors were determined to be 0.5 µM and 0.04 µM, respectively. Furthermore, selectivity measurement results revealed that the proposed non-enzymatic biosensor has remarkably good sensitivity and high selectivity, indicating that common biomolecules (glucose, maltose, lactose, fructose, and urea) could be captured by the sensor. Consequently, it was proven that the proposed biosensor could be a potential device for diabetes diagnosis.