The sanctions will freeze any US-based assets and ban any financial transactions with the targets, who include Vladimir Zaritsky, the former commander-in-chief of Russia's missile forces and artillery who is leading a hotel project in Crimea.
The US Treasury Department "remains committed to targeting Russian-backed entities that seek to profit from Russia's illegal annexation and occupation of Crimea," said Sigal Mandelker, the Treasury under secretary for terrorism and financial intelligence.
"Our sanctions are a clear reminder that efforts seeking to normalize investment and economic relationships with those operating in Crimea will not be tolerated and are subject to US and EU sanctions authorities," she said in a statement.
Western powers were effectively powerless to stop the annexation but have vowed never to recognize the move.
Also under the new sanctions is the Mriya Resort and Spa, a luxury hotel that opened in the resort of Yalta shortly after the annexation and which the Treasury Department called "the main Russian platform for showcasing investment opportunities in Crimea."
The hotel's website says that it includes a medical center and is designed based on a concept of British architect Norman Foster, who built Wembley Stadium.
The Treasury Department also imposed sanctions on two people over alleged human rights abuses related to Russia's role in Ukraine.
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