In recent years, fungal infections have emerged as a significant health concern across veterinary species, especially in livestock such as cattle, where fungal diseases can result in considerable economic losses, as well as in humans. In particular, 
Aspergillus
 species, notably 
Aspergillus flavus
 and 
Aspergillus versicolor
, are opportunistic pathogens that pose a threat to both animals and humans. This study focuses on the synthesis and antifungal evaluation of novel 9-fluorenylmethoxycarbonyl (Fmoc)-protected 1,2,4-triazolyl-α-amino acids and their dipeptides, designed to combat fungal pathogens. More in detail, we evaluated their antifungal activity against various species, including 
Aspergillus versicolor
 (ATCC 12134) and 
Aspergillus flavus
 (ATCC 10567). The results indicated that dipeptide 
7a
 exhibited promising antifungal activity against 
Aspergillus versicolor
 with an IC
50
 value of 169.94 µM, demonstrating greater potency than fluconazole, a standard treatment for fungal infections, which showed an IC
50
 of 254.01 µM. Notably, dipeptide 
7a
 showed slightly enhanced antifungal efficacy compared to fluconazole also in 
Aspergillus flavus
 (IC
50
 176.69 µM vs. 184.64 µM), suggesting that this dipeptide might be more potent even against this strain. Remarkably, 
3a
 and 
7a
 are also more potent than fluconazole against 
A. candidus
 10711. On the other hand, the protected amino acid 
3a
 demonstrated consistent inhibition across all tested 
Aspergillus
 strains, but with an IC
50
 value of 267.86 µM for 
Aspergillus flavus
, it was less potent than fluconazole (IC
50
 184.64 µM), still showing some potential as a good antifungal molecule. Overall, our findings indicate that the synthesized 1,2,4-triazolyl derivatives 
3a
 and 
7a
 hold significant promise as potential antifungal agents in treating 
Aspergillus
-induced diseases in cattle, as well as for broader applications in human health. Our mechanistic studies based on molecular docking revealed that compounds 
3a
 and 
7a
 bind to the same region of the sterol 14-α demethylase as fluconazole. Given the rising concerns about antifungal resistance, these amino acid derivatives, with their unique bioactive structures, could serve as a novel class of therapeutic agents. Further research into their in vivo efficacy and safety profiles is warranted to fully realize their potential as antifungal drugs in clinical and agricultural settings.