Khamenei's son, Mojtaba Khamenei, is an influential religious and political figure in the country, with European sources previously mentioning his name among the possible successors of his father as Iranian supreme leader.
Trump wants to force Iran to renegotiate the 2015 nuclear accord, arguing that the terms were not tough enough to prevent the country from developing nuclear weapons, agree curbs to its ballistic-missile program, and end its "malign" activities in the Middle East.
"Kneeling before the enemies and obeying them will never have any achievements for the Iranian nation and the Islamic Revolution", General Mousavi added in an address to the public rallies in Tehran on Monday, which are being held to mark the 40th anniversary of the United States embassy takeover in Tehran.
Four decades to the day after revolutionary students raided the embassy and seized dozens of United States diplomats and security staff, demonstrators in Tehran and several other cities shouted anti-American slogans and flaunted effigies mocking President Donald Trump.
Iran has announced a tenfold increase in enriched uranium production as Tehran backs away from its nuclear deal with the west. It allowed Iran to refine uranium only with first-generation IR-1 centrifuges.
Macron's efforts to initiate a phone call between US President Donald Trump and his Iranian counterpart Hassan Rouhani on the sidelines of the UN General Assembly in September ended in failure.
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The announcement also said Iran now has a prototype centrifuge that works 50 times faster than those allowed under the deal.
Among the events planned in Tehran on Monday is a rally by hard-liners at the former embassy and an address by Iranian army commander Gen. Abdolrahim Mousavi.
This year's commemoration of the embassy seizure comes as Iran's regional allies in Iraq and Lebanon face widespread protests.
Meanwhile Monday, demonstrators gathered in front of the former US Embassy in downtown Tehran as state television aired footage from other cities across the country making the anniversary. Violence there killed three people and wounded 19, Iraqi officials said. News agencies posted images of protesters setting the US flag on fire.
'Americans, at the time (of the embassy takeover) believed they were an exceptional nation, able to commit any injustice anywhere, ' government spokesman Ali Rabiei told the press on Monday.
Ayatollah Khamenei's tirade about Washington came on the eve of the 40th anniversary of the seizure of the USA embassy in Tehran.
But since the deal collapsed, European nations have been unable to give Iran a way to help it sell its oil overseas as it faces renewed US sanctions. Tehran denied the allegation, though it did seize oil tankers and shoot down a USA military surveillance drone.