The sanctions will freeze any US-based assets and ban 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.
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"Our sanctions are a clear reminder that efforts seeking to normalise 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.
In 2014, Moscow annexed Crimea, the Black Sea peninsula whose population is largely ethnically Russian but was part of Ukraine, as part of a struggle to keep Kiev from falling into the Western orbit.
Western powers were effectively powerless to stop the annexation but have vowed never to recognise the move.
The measures come as the State Department says it is preparing wider sanctions against Russia, as required by Congress, over a nerve attack carried out in Britain against a former double agent.
Russia has hit back over those sanctions and vowed reciprocal measures.
Despite the sanctions, President Donald Trump has described an affinity for his Russian counterpart Vladimir Putin. The two will both attend November 11 ceremonies in Paris for the centennial of the end of World War I and then rub shoulders in Buenos Aires at the Group of 20 summit.
The United States also sanctioned Zaritsky's firm for buying three hotels in Crimea that had been owned by the Ukraine state and were taken over after the annexation.
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 additionally imposed sanctions on two people over human rights issues related to Russia's role in Ukraine.
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They are Andriy Volodymyrovych Sushko, identified as an officer of the FSB security agency, who allegedly abducted and tortured an activist from Crimea's Tatars, a minority group that was persecuted under Stalin and has seen increasing pressure since Moscow returned to the peninsula.
Also sanctioned was Alexander Basov, the deputy ministry of state security in the internationally unrecognised Luhansk People's Republic in eastern Ukraine, where more than 10,000 people have been killed since 2014 in a Russian-backed insurgency following the annexation of Crimea.
The US Treasury Department said Basov has used his position to persecute Jehovah's Witnesses, the heterodox Christian group known for proselytisation which human rights groups say has faced growing exclusion in Russia.
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