Ben Te'o has been left out of the England squad for Sunday's international against Wales.
The decision to exclude the inside centre casts further doubt on him being named in the final 31-man squad for the World Cup on Monday.
Mike Brown, Alex Dombrandt and Ben Spencer were other notable absentees from the 33-strong line-up, who were gathering at Pennyhill Park on Tuesday. No new names have been added to the squad following England's 12-day training camp in Treviso.
Head coach Eddie Jones insisted it was not the end of the road for those left out, saying: "Players excluded from being involved in this camp are naturally disappointed but may get another opportunity so they must be ready."
England will play Wales twice as well as Ireland and Italy before flying to Japan ahead of their first World Cup match against Tonga on September 22.