The US measures came as Venezuela's opposition began a two-day nationwide strike aimed at ousting the president through early elections.
The deadliness of four months of violent anti-Maduro protests was further confirmed with the death of a 30-year-old man in a demonstration in the west of the country. That added to a death toll that has already surpassed 100.
In Washington, the US Treasury unveiled a list of 13 current and former officials, including the interior minister, senior military brass, the president of the electoral council, and the finance chief of state oil company PDVSA, whose US assets would be frozen.
Critics say the body is a step towards a dictatorship, by bypassing or dissolving the opposition-held National Assembly.
Maduro called the US punishment "illegal, insolent and unprecedented."
"Who do these imperialists in the United States think they are? The government of the world?" he said in a speech.
But in his country, where there are widespread shortages of basic goods and soaring inflation, protesters are showing their discontent with Maduro's leadership.
"No more dictatorship!" read signs on road barricades in eastern Caracas.
The Venezuelan military has declared its loyalty to him.
But some 70 percent of Venezuelans are opposed to the Constituent Assembly, according to polling firm Datanalisis.
The hardening political struggle has deepened fears that months of street violence could worsen.
The opposition has planned another major demonstration in the capital on Friday.
Thousands of Venezuelans loaded with heavy bags have crossed the border into Colombia this week, fleeing the unrest.
"The elections are on Sunday and we really don't know what will happen," said one, Maria de los Angeles Pichardo, who left with her husband and son. "To be safe, we prefer to cross."
"Every time we're worse off, with long lines and shortages. I think I'll strike for 48 hours," said one Caracas resident, Maria Auxiliadora.
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