"There is nothing to mediate," Foreign Minister Adel al-Jubeir told reporters.
"Canada made a big mistake... and a mistake should be corrected."
Jubeir added that Saudi Arabia was "considering additional measures" against Canada, without elaborating.
Saudi Arabia on Monday expelled Canada's ambassador and recalled its own envoy, freezing all new trade and investments, after Ottawa denounced a new crackdown on rights activists in the kingdom.
Riyadh also said it will relocate thousands of Saudi students studying in Canada to other countries, while state airline Saudia (also known as Saudi Arabian Airlines) announced it was suspending flights to Toronto.
Saudi Arabia has also stopped all medical treatment programs in Canada and is working on transferring all Saudi patients there to other countries, state media said Wednesday.
Further straining ties, the Saudi central bank has instructed its overseas asset managers to dispose of their Canadian equities, bonds and cash holdings "no matter the cost", the Financial Times reported on Wednesday.
Canada sparked fury in Riyadh last week when it called for the "immediate release" of rights campaigners, including award-winning women's rights activist Samar Badawi, the sister of jailed blogger Raif Badawi.
Samar Badawi was arrested along with fellow campaigner Nassima al-Sadah last week, the latest targets of what Human Rights Watch called an "unprecedented government crackdown" on women activists.
It came after more than a dozen women's rights campaigners were detained and accused of undermining national security and collaborating with enemies of the state.
When asked about the jailed activists, Jubeir reiterated the government's earlier stance that they had been in contact with foreign entities, but did not specify the charges against them.
"The matter is not about human rights, it is a matter of national security," Jubeir said, referring to the row with Canada.
"Saudi Arabia does not interfere in the affairs of Canada in any way. Therefore, Canada must correct its actions towards the kingdom."
Canada has indicated it will not back down, despite the risk of imperilling business deals including a $15 billion (13 billion euros) agreement to sell Riyadh light armoured vehicles.
If the deal is scrapped, thousands of jobs could be lost in Canada, experts say.
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