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Drugs play a key role in the treatment of patients with various diseases. A compound, when administered systemically, shows differential spatial and temporal distribution patterns not only in the body but also within each organ. In response to an increase or decrease in local concentrations in the organ, the activity of the cell population expressing the drug’s target protein(s) changes over time. Therefore, real time, simultaneous detection of kinetics of the drug and its pharmacological effects in in vivo microenvironments is essential for evaluating the efficacy of medicines. Although such challenging dual-mode measurement has not yet been addressed by any conventional methods, it has been successfully achieved via a microsensing system that we recently developed. The system consists of two different sensors: A needle-type boron-doped diamond microelectrode for monitoring the drug and a glass microelectrode for tracking electrophysiological activity of the target cells. This state-of-the-art approach is applicable to various drugs in terms of “local” pharmacokinetic and pharmacodynamic assays in vivo and may contribute to the development of next-generation therapeutic interventions.