Protestors in the Spanish capital marched in support of Jenni Hermoso nine days after Rubiales - the president of the Spanish Football Federation (RFEF) - kissed the Spain attacker without consent during the medal ceremony following the Women's World Cup final.
Protestors at the march, organised by the Comision8M feminist movement, carried banners bearing slogans including "Rubiales resign", "Sexists and gentlemen, sports are for everyone" and "Out with macho federations."
Rubiales has been suspended for 90 days by FIFA after he refused to resign during an extraordinary press conference on Friday.
The Spanish government has asked Spain's Sports Tribunal (TAD) to suspend Rubiales, while the RFEF threatened Hermoso with legal action after Rubiales claimed that she initiated the kiss.
On Monday it emerged that Rubiales' mother had begun a hunger strike at a church in Motril because of what she described as an "inhuman hunt' against her son.
The Spanish women's national team coach Jorge Vilda eventually criticised the behaviour of close friend Rubiales, but only after his entire coaching staff had resigned and 81 players ruled themselves out of representing Spain until change is implemented.
The events of the past week have been dubbed Spain's MeToo movement, with many sensing an opportunity to rid the country's sporting and public institutions of institutional sexism and machoism.