Rapid in vitro and computer-aided method for assessing synergistic interactions between NSAIDs and analgesics

Pain is a complex phenomenon that plays a significant role in various diseases, influencing both the physical and psychological well-being of individuals. In clinical practice, combining nonsteroidal anti-inflammatory drugs (NSAIDs) with analgesics, such as paracetamol or metamizole, has become a widely adopted strategy to manage pain. Although the synergistic effects of combining NSAIDs with analgesics are well recognized in clinical practice, this approach is primarily based on empirical clinical experience. Our work aims to present a rapid method for evaluating the anti-inflammatory effects of drug combinations through in vitro assays combined with computer-aided data processing and analysis. We conducted two simple and rapid in vitro assays, the Griess and DPPH assays, to evaluate the effects of NSAID-analgesic combinations and demonstrate their synergistic interactions, using the free web application SynergyFinder Plus. This computer-aided analysis enabled a quantitative assessment of drug interactions, enhancing the interpretation of the experimental data. Furthermore, to better understand the results obtained from previous experiments, we analysed the anti-inflammatory effects of ketoprofen and dexketoprofen in combination with metamizole and paracetamol through quantitative real-time PCR (qRT-PCR). Our findings reveal synergistic interactions between NSAIDs and analgesics in terms of their anti-inflammatory and antioxidant activities. This work could be the first step for the study of the mechanisms behind the synergistic interactions between NSAIDs and analgesics for the treatment of pain, mainly when inflammatory processes are involved. Consequently, this study aims to contribute to the exploration of non-opioid drug combinations, addressing the urgent need for alternative analgesic strategies that minimize opioid use.