The State Department said Yehia Sinwar and Rawhi Mushtaha along with the Palestinian group's military commander Mohammed Deif had been named "specially designated global terrorists" under US law.
Deif has been described as Israel's most wanted man for his role since 2002 as commander of Hamas' military wing, the Ezzedine al-Qassam Brigades, overseeing attacks in Gaza and Israel.
"He is known for deploying suicide bombers and directing the kidnapping of Israeli soldiers," the State Department said in a statement announcing his new designation.
As "global terrorists," the three Palestinian figures are subject to the seizure of any assets they hold in areas of US jurisdiction and Americans are forbidden from doing business with them.
The Palestinian Islamist movement Hamas, which controls the Gaza Strip enclave, is regarded as a terrorist organisation by both the United States and Israel.
The Al-Qassam Brigades are its armed wing and regularly fire rockets into Israel.
Deif, a veteran of the Palestinian struggle, has long been a target for Israeli assassination, and in August last year his wife and son were killed in an air strike.
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