CD73 inhibitors are considered to be used in the therapies of melanomas, gliomas or breast cancer. However, little is known about their pharmacology and kinetics in mouse experimental models. Thus, this study is aimed to define a metabolic stability and elimination of the adenosine diphosphate (ADP) analog - α,β-Methylene-ADP also known as AOPCP in BALB/c mice. The process starts with an intravenous injection of AOPCP, next blood and serum samples are collected. Urine samples are possessed by a bladder puncture. Mice aortas are dissected for the e5NT activity evaluation. In order to assess the AOPCP degradation, the incubation of AOPCP in mice blood and plasma is performed. The AOPCP concentration as well as the activity of e5NT were analyzed with the reverse phase-high pressure liquid chromatography (RP-HPLC). The study shows that after 60 minutes of the 20 mg/kg intravenous injection of AOPCP (body weight dose), the concentration of AOPCP in blood diminished rapidly from 38.6 ± 5.0 µM (measured 5 minutes after the injection) to 6.4 ± 1.4 µM. Interestingly, it is also noted that 60 minutes after the incubation of mice blood samples the AOPCP concentration decreases from 50 µM to 30.0 ± 0.3 µM. This study demonstrates a significant and quick decrease of AOPCP concentration in BALB/c mice blood after the intravenous injection and in isolated blood sample incubation. These findings emphasize the quick elimination of AOPCP as well as its instability and suggest that the AOPCP concentration have to be accurately and frequently monitored in all the studies that address its clinical application.