Hops are widespread as a wild plant in almost all Northern and Central Italy, and the characterization of wild populations is attracting considerable interest in verifying their potential use. The development of hops as agricultural crop can be an interesting opportunity, both for farms that would have available a new crop to be included in the crop system and for craft breweries interested in characterizing beers with local raw materials. In the present work, 14 wild hop accessions coming from various Italian locations were characterized and compared with 2 commercial varieties (Cascade and Hallertau Taurus) grown in the same environments. The cones were analyzed to measure the content of α- and β-acids, polyphenols, flavonoids, and the anti-radical power. The α-acid content of wild hops was generally low, while the β-acid content was very variable and quite high in some samples. The content in polyphenols and flavonoids and the antiradical power were high and generally similar to those of the commercial varieties. Therefore, the analyzed genotypes are not very suitable for use as bitter hops in beer production, while further analysis may indicate a possible use as aroma hops, or for herbal and pharmaceutical purposes, thanks to their antioxidant content.