Brad Barritt and Tom Croft will play no part in the rest of England's Six Nations campaign after picking up injuries on club duty.
Centre Barritt has not featured on the international stage since the victory over Australia last November due to calf and knee injuries.
However, the Saracens man was set to be in contention for Saturday's Calcutta Cup clash against Scotland before hurting his ankle in last weekend's victory over Wasps.
Leicester Tigers flanker Croft is also unavailable after dislocating his shoulder during the 16-12 win over Newcastle Falcons.
Squad member Henry Thomas, who made the last of his seven Test appearances in June, is also ruled out through injury.
There was better news for England, though, as Mike Brown, Tom Wood and Courtney Lawes should all be fit for the match at Murrayfield this weekend.
Stuart Lancaster's side finish off their campaign against France at Twickenham a week on Saturday.