Pyrazolines have played a crucial role in the development of various biologically active compounds. At the same time, the ferrocene moiety represents a significant part of the structure of these types of molecules. For this reason, we presumed that the incorporation of different pharmacophores in the same structure could lead to interesting changes, and we decided to synthesize some new ferrocenyl N-acyl pyrazolines. The DNA protective effect of four selected ferrocenyl N-acyl pyrazolines namely 5-methyl-3-ferrocenyl-4,5-dihydro-1H-pyrazole-1-carbaldehyde (3b), 1-(5,5-dimethyl-3-ferrocenyl-4,5-dihydro-1H-pyrazol-1-yl)ethanone (4c), 1-(5-(furan-2-yl)-3-ferrocenyl-4,5-dihydro-1H-pyrazol-1-yl)ethanone (4h), and 1-(3-ferrocenyl-4,5-dihydro-1Н-pyrazol-1-yl)propan-1-one (5a) in concentration of 100 μg/mL against hydroxyl and peroxyl radicals-induced DNA damage was assessed using salmon sperm DNA sodium salt as a model system. The acridine orange assay was performed to analyze the DNA integrity of the selected compounds at the same concentrations. The tested compounds in the selected dose had a statistically significant potency to protect DNA from damage caused by hydroxyl and peroxyl radicals. According to the acridine orange assay selected compounds significantly reduced hydroxy and peroxyl radicals induced denaturation and damage to DNA. The results showed that the tested compounds showed strong protective activity against hydroxy and peroxyl radicals-induced DNA damage.