Marcus Rashford, who is in fine form for Manchester United and will miss England's Euro 2024 qualifiers against Italy and Ukraine, suffered an injury during Sunday's FA Cup victory against Fulham.
Chelsea midfielder Mason Mount and Newcastle goalkeeper Nick Pope have left Gareth Southgate's team.
Southgate, who selected a 25-man roster last week, has only called up Tottenham goalkeeper Fraser Forster as a replacement.
The most significant loss is Rashford's absence, who has been on fire since the World Cup.
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For United this season, the 25-year-old has a career-high 27 goals, 19 of which have come since his return from international service in Qatar, where he scored three goals in five games.
Mason Mount of Chelsea and goalkeeper Nick Pope of Newcastle have also withdrawn from Gareth Southgate's team.
Southgate, who selected a 25-man roster last week, has only called up Tottenham goalkeeper Fraser Forster as a replacement.
The most significant loss is Rashford's absence, who has been on fire since the World Cup.
At Manchester United this season, the 25-year-old had scored a career-high 27 goals, 19 of which had come following his return from international service in Qatar, when he scored three goals in five games.
In the 83rd minute, Rashford was replaced by Erik ten Hag with Manchester United leading 2-1 after a tumultuous closing 20 minutes of the FA Cup quarterfinal between Manchester United and Fulham. Since then, it has been established that the 25-year-old sustained an injury during the eventual 3-1 victory and has been forced to miss England's forthcoming pair of matches as a result.
When club football resumes in April, United claim on their website that Rashford will 'hopefully be available' for their away trip to Newcastle, while teammates Harry Maguire and Luke Shaw will continue to represent England as expected.