Amauri, never previously considered by his native Brazil, was earlier given a late call-up following an injury to Luis Fabiano but Juve had the final say because the FIFA deadline for selecting overseas players for the match had passed.
"The club has decided not to accept the request of the Brazilian federation," Juve president Giovanni Cobolli Gigli told Sky after his side's 3-2 Serie A defeat by Cagliari.
"We need to stay concentrated on the job in hand, now more than ever."
The 28-year-old Amauri, who signed from Palermo in the close season, is applying for Italian citizenship and has been undecided for months about which country to represent.
Cobolli Gigli, who has yet to receive a formal request from Brazil, said the player's views had not entered into the club's decision.
Brazil face Italy on February 10 at Arsenal's Emirates Stadium in London.
Italy boss Marcello Lippi has said he will consider selecting Amauri in the future but the World Cup winning-coach has become frustrated at the time it is taking for the powerful forward's Italian passport to come through.
