The victory at Mestalla was the first match of the season Karim Benzema, Gareth Bale and Cristiano Ronaldo started together and the return of the 'BBC' strikeforce could limit Isco's involvement over the second half of the campaign.
Asensio may also be pushed down the pecking order even though he has scored four La Liga goals - double Benzema's haul despite starting three league matches fewer.
While Isco and Asensio's game time is a concern for Lopetegui with the World Cup approaching, the form of Diego Costa at Atletico Madrid is a positive, the Brazil-born forward having scored three goals in his five appearances since returning from Chelsea.
"I'm sure they will play again, but I'm worried that Marco and Isco play little," Lopetegui told reporters at the Spanish Sports Press Awards.
"Diego Costa has not surprised me and he can be in the next squad."
Royal Spanish Football Federation president Juan Luis Larrea, also speaking at the event, confirmed Lopetegui is set to sign a new Spain contract.
"The renewal of Julen is underway," Larrea said.
"And it is very likely that it will be closed soon."