Barcelona has failed in its appeal of Neymar's three-match suspension that rules him out of El Clasico on Sunday against LaLiga leaders Real Madrid.
Neymar was handed the punishment for sarcastically applauding the fourth official after being shown a second yellow card in the 2-0 loss at Malaga earlier this month.
The Brazilian forward's show of dissent earned him an additional two-match suspension on top of the one-game ban that came as a result of the red card at La Rosaleda, ruling him out of the clash with Madrid at Santiago Bernabeu this weekend and the meeting with Osasuna next week.
Barca argued it could not be proved Neymar's reaction was directed at the official, but the Royal Spanish Football Federation's Appeals Committee rejected that claim and upheld the sanction.
But Barca could yet take its appeal to the Court of Arbitration for Sport in order to have his punishment overturned.
The news will come as a blow for Luis Enrique in the wake of his side's Champions League elimination at the hands of Juventus on Wednesday.
Defeat at the Bernabeu would severely damage the Catalans' title ambitions, with Madrid holding a three-point advantage at LaLiga's summit and with a game in hand.
Barca will already be without long-term absentees Aleix Vidal and Rafinha, while Arda Turan's fitness continues to be monitored.
Should the club fail in a prospective appeal to CAS, Neymar would return for the derby against Espanyol on April 29.