The Portugal international was sent off in the 1-1 draw with Athletic Bilbao earlier this month, having appeared to strike Carlos Gurpegi in the face following an altercation between the pair.
Real appealed the Spanish Football Federation's three-match ban and, when this was rejected, took their case to the Ministry for Education, Culture and Sport - who also dismissed the case.
Carlo Ancelotti expressed his frustration at the lengthy appeal process on Friday, and claimed he had included Ronaldo in his match-day squad, hinting at a confidence the ban would be overturned.
Having already missed league clashes with Villarreal and Getafe, Ronaldo will now serve the final game of the original ban when Madrid take on Elche.
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