Washington said those targeted were “leaders, financiers, and facilitators” of ISIS in Yemen and al-Qaeda in the Arabian Peninsula.
The sanctions list was jointly agreed by Saudi Arabia, Qatar, the United Arab Emirates, Oman, Kuwait and Bahrain.
US Treasury Secretary Steven Mnuchin branded the measures “the largest ever multilateral designation in the Middle East”.
Mr Mnuchin said: “Through this action we are aggressively targeting radical extremists in Yemen and the surrounding region who pose a direct threat to the security of the United States, Yemen, and the international community.
“This bold and innovative multilateral approach is needed because terrorism poses a threat to all of our nations.
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“It is critical that we come together to combat this.”
The announcement, made in the Saudi capital, represents a rare moment of cooperation among the oil-rich Gulf states.
Four months ago, Saudi Arabia, the UAE and Bahrain, along with Egypt, severed ties with Qatar over its foreign policies.
Mr Mnuchin is scheduled to visit the UAE, Qatar and Jerusalem during the rest of trip to the Middle East.
He is expected to discuss opportunities to collaborate with US allies to tackle “threatening activities” from Iran.
He told a press conference in Ridyah: “This will include a focus on taking aggressive actions against Hizballah, whose main benefactor is Iran.”
US President Donald Trump first announced his plan to target ISIS’ alleged financial backers in May.
He said: “As we deny terrorist organisations control of territory and populations, we must also strip them of their access to funds.
“We must cut off the financial channels that let ISIS sell oil, let extremists pay their fighters, and help terrorists smuggle their reinforcements.
“There is still much work to do. That means honestly confronting the crisis of Islamist extremism and the Islamist terror groups it inspires.”