The sweet chestnut fruit has always had great importance in the southern European countries. Chestnut production is an important source of income and a crop of high environmental value thanks to its role in soil protection. It is also a good food with enormous potential for various aspects of health because of its nutritional qualities. The quality of sweet chestnuts is affected by various factors, such as climatic conditions and cultivation inputs. It is very important to recognize the impacts of climate on chestnut fruits, to improve our current understanding of climate–chestnut interconnections. The current study investigated and compared the antioxidant activity and the total phenolic and flavonoid contents of different cultivars of chestnuts grown in different geographic areas of northwest Spain. The results obtained with three antioxidant capability assays (DPPH, ABTS and FRAP assays) were highly correlated. All the samples had high antioxidant capacity and high total phenolic and total flavonoid contents, which depended both on cultivar and growth region. Ventura variety, harvested in the coldest environments, presented the highest values of antioxidant activity (IC50DPPH = 34.5 g/L), total phenolic content (131.84 mg equivalent of gallic acid/100 g FW) and total flavonoids (7.77 mg eq. catechin/100 g). The variations in the antioxidant capacity, total phenolic and total flavonoid contents of different cultivars, and their associations with climatic environmental factors, revealed the significant impacts of these factors on the synthesis of specialized metabolites and on the nutraceutical potential of chestnuts. The results can provide valuable information for selection of the cultivar and the cultivation conditions of the chestnut, in order to obtain chestnuts with high-quality bioactive characteristics.