As "Royal Charles" she took part in the Second Anglo-Dutch War. In 1665, she fought in the Battle of Lowestoft under the command of the Lord High Admiral, James Stuart, Duke of York, her captain being Sir William Penn. During that battle she probably destroyed the Dutch flagship "Eendracht". In 1666, she participated in two further actions, the Four Days Battle and the defeat of Admiral Michiel de Ruyter in the St. James's Day Battle off the North Foreland.