Objectives: This study aimed to evaluate the effects of 5-butyl-2-pyridinecarboxylic acid on growth inhibition, biofilm formation, biofilm destabilization, and cellular integrity of four different Candida species. Materials and Methods: The anti- Candida activity of 5-butyl-2-pyridinecarboxylic acid was assessed using disk diffusion and microdilution assays to determine minimum inhibitory concentration (MIC) and minimum fungicidal concentration (MFC). Growth kinetics were monitored through growth curve and time-kill assays, while synergistic effects with standard antifungal drugs were evaluated using fractional inhibitory concentration (FIC) indices. Cellular damage was analyzed by scanning electron microscopy (SEM) and lactate dehydrogenase (LDH) release assays. Biofilm inhibition and destabilization were determined using microtiter plate assays with crystal violet staining. A molecular docking study predicted the potential target of action. Results: The compound exhibited potent inhibitory activity, with MIC and MFC values ranging from 0.129 ± 0.013 to 0.258 ± 0.012 mg/ml. At concentrations between 0.128 and 0.258 mg/ml, it induced rapid viability loss and complete lethality within 15 hours. SEM and LDH assays revealed significant cell wall disruption and increased permeability. Time-kill kinetics confirmed potent fungicidal activity. The compound also showed synergistic effects with clotrimazole and luliconazole against C. albicans and C. glabrata, while partial synergy was observed against C. parapsilosis. Moderate antibiofilm activity was also recorded compared to usnic acid. 5-butyl-2-pyridine carboxylic acid was predicted as a potential target of Candida ergosterol biosynthetic enzyme 14α-demethylase (CYP51). Conclusions: This study highlights the promising anti- Candida potential of 5-butyl-2-pyridine carboxylic acid. By disrupting cell walls, increasing membrane permeability, exhibiting synergism with conventional antifungals, and being predicted as an inhibitor against 14α-demethylase (CYP51), the compound represents a potential lead candidate. Further safety and mechanistic studies are needed to explore its application in antifungal drug development.