Thomas Tuchel's team were unable to hold off a late rally from Luis de la Fuente's side at Wembley and the Three Lions squad are in need of a boost on the road in Prague on September 29th.
Czechia also lost out in their first game, going down 2-1 at home to Croatia, and the away side are expected to rotate to avoid game overload due to four matches within a short timeframe.
England v Czechia team news
Ezri Konsa and Nico O'Reilly both missed England's first training session after the Spain match and Tuchel could leave them out as a precaution.
Jarrad Branthwaite and Lewis Hall could be drafted into the starting XI with Myles Lewis-Skelly backed to keep his place in midfield.
Jordan Pickford will come back in as No.1 with the recalled Trent Alexander-Arnold also pushing to start at right-back.
England predicted lineup v Czechia
England XI: Pickford; Alexander-Arnold, Branthwaite, Guehi, Hall; Anderson, Bellingham, Lewis-Skelly; Saka, Kane, Gordon.
England v Czechia kick off time
Kick off at the Fortuna Arena is set for 8:45pm local time and 7:45pm in the UK.
Key stat
England famously beat Czechia on route to the final of Euro 2020, but they have lost their sole game away in Prague, after going down 2-1 in 2019.