Saudi Arabia, the United Arab Emirates and Bahrain have issued a joint statement withdrawing their ambassadors from Qatar.
"The three states have decided to start taking measures they see fit to protect their security and stability, affirming that they are safeguarding the interests of the Gulf Cooperation Council (GCC) states, including neighbouring Qatar," the statement read.
Qatar has failed to "stick to the principles of non-intervention in the internal affairs of other states and refraining from supporting all who threaten the security of the council states," the statement added.
In response, Qatar said it "regrets" the move, but will not reciprocate, AFP reported
Qatar's emir Sheikh Tamim bin Hamad Al-Thani committed to a security agreement during a meeting with Kuwait's emir and the Saudi monarch in Riyadh on 23 November 2013.
Calls for Qatar to abide by the agreement were reiterated on 17 February in Kuwait and again during a meeting of foreign ministry heads in Riyadh on 4 March.
Qatar has failed to take the proper measure to put the agreement into effect, according to Wednesday's statement.
The statement additionally cited Qatar's support for "antagonistic media," a reference to the Qatari-owned Al Jazeera satellite channel. The channel has been accused of providing biased coverage in favour of the Muslim Brotherhood.
The trial of 20 journalists for supporting the group -- which has been officially labelled a terrorist organisation by the Egyptian authorities -- is to resume in Cairo on Wednesday.
Saudi Arabia and other Gulf monarchies have been fearful that the Brotherhood's fall in Egypt could shake their authority in their own countries.
The UAE has detained Emirati and Egyptian Islamists for alleged links to the group. According to AFP, a Qatari national was sentenced to seven years in a UAE prison for fundraising for a local Brotherhood-linked group.
Qatar is a major backer of Syrian rebels, alongside Saudi Arabia.