[2] Seventeen teams qualified for the 2022–23 EPCR Challenge Cup from Premiership Rugby, the Top 14 and the United Rugby Championship as a direct result of their domestic league performance having not qualified for the Heineken Champions Cup.

The knockout stage was played across 31 March/1/2 April with a single leg round of 16 matches consisting of the top six ranked teams from each pool and the teams ranked 9th and 10th in each pool of the 2022–23 European Rugby Champions Cup (Clermont, Lyon, Racing 92 and Sale Sharks denoted by * in the bracket).

The Round of 16 followed a pre-determined format, while the quarter-finals and semi-finals always guaranteed home advantage to the higher ranked team.

The higher-ranked club gained home stadium advantage, however, in the event that a South African team had been the higher ranked side, the game would have been located in Europe.

Note: Flags to the left of player names indicate national team as has been defined under World Rugby eligibility rules, or primary nationality for players who have not yet earned international senior caps.