Teams will be allowed to make five substitutions from nine players on the bench for the EFL Cup semi-finals and final.
The change in the rule was introduced following consultation with Manchester United, Manchester City, Tottenham and Brentford, the four semi-finalists in the 2020-21 competition.
VAR will also be introduced, having not been involved in previous rounds.
The EFL added in a statement on Monday: "Each team is only permitted a maximum of three opportunities (excluding half time, full time and half time of extra time) to make substitutions during normal time. However, if a tie goes to extra time, an additional opportunity to use a substitute(s) will also be permitted but only from the five substitutes allowed."
The decision comes despite Premier League clubs voting against the option of using five substitutes in a single game in England's top flight, a rule which has been adopted in other major competitions across Europe this season.
City boss Pep Guardiola has been among the more vocal in his criticism of not continuing the five-sub rule, which was temporarily introduced into the Premier League following the resumption of the 2019-20 season after the coronavirus-enforced shutdown.
City face United at Old Trafford on Wednesday, with Spurs hosting Brentford 24 hours earlier.
This season's semi-finals will be one-legged and go to extra time and penalties if required, rather than the traditional two-legged ties, due to the congested fixture schedule caused by the disruption to the football calendar.
The final was moved from February to April, when it is hoped some fans may be allowed to attend.