The Premiership Rugby clubs will face the RFU Championship sides - with the competition kicking-off on September 8, 2023.
The 24 teams will be split into four pools of six - based on league standings at the end of the current 2022/23 season.
Phil Winstanley, Premiership Rugby’s Rugby Director said: "It is many years since the Premiership and Championship Clubs clashed in a Cup competition.
"The Championship performs a crucial role within the English game and we’re looking forward to renewing some rivalries of old.
"With the Rugby World Cup taking place in France, this competition will no doubt provide the best, young English talent with an opportunity to stake a claim in squads before the start of the Gallagher Premiership Rugby campaign."