England trio Henry Slade, Sam Underhill and Ruaridh McConnochie have been ruled out of the World Cup warm-up game against Wales at Twickenham on Sunday.
Wing Ruaridh McConnochie was set to make his England debut but has a hip injury.
Flanker Underhill and centre Slade were also expected to start but have respective knee and toe problems.
Joe Cokanasiga, Jonathan Joseph and Lewis Ludlum will now start in the first of four warm-up games.
Northampton flanker Ludlam was originally due to make his debut off the bench.
England sevens star McConnochie only switched to the XV-a-side version last summer.
His absence against Wales will mean he misses a chance to impress before England head coach Eddie Jones names his final 31-man World Cup squad on Monday.
Meanwhile, Jones has refused to confirm a report in The Times that an altercation took place between Mike Brown and Ben Te'o during the side's training camp in Treviso.
The Rugby Football Union admitted an incident did happen in Treviso.
Jones said: "No I'm not able to confirm that. I'm not able to confirm. I think I've answered the question."