Chelsea duo John Terry and Ashley Cole will both miss England's friendly international against France, with Aston Villa's Stephen Warnock and Bolton Wanderers' Gary Cahill called up as replacements.
Terry missed the shock 3-0 defeat at home to Sunderland, having struggled with a hamstring niggle during training.
Blues boss Carlo Ancelotti confirmed:"Terry trained yesterday but at the end of the training session, he had pain on his nerve in his hamstring and we had the test this morning.
"He never missed a training session, but yesterday the pain was more, so he was not able to play."
Left-back Cole suffered an ankle knock against the Black Cats, but is not thought to be out for a matter of days.
Meanwhile, West Ham United striker Carlton Cole has been added to Fabio Capello's squad as cover for Andy Carroll and Gabriel Agbonlahor.
Newcastle's Carroll and Villa's Agbonlahor both picked up unspecified injuries for their clubs on Saturday, with the latter definitely ruled out of Wednesday's international.
Cole won the last of his seven England caps as a second-half substitute against Egypt in March.