The maverick cleric had last week condemned the suspected deadly use of chemical weapons by Assad's forces against civilians, becoming a rare Shiite leader to openly challenge the Syrian president's legitimacy.
Sadr issued a new statement today that reiterated his position.
"I have urged him to step down to preserve the reputation of the Mumanaa and to escape a Gaddafi fate," he said, using a word that refers to a so-called anti-Western "resistance front" that includes Lebanon's Hezbollah, Iran and Syria.
A chemical attack which has been widely blamed on Assad's regime killed 87 civilians, including 31 children, in the rebel-held Syrian town of Khan Sheikhun on April 4.
The United States subsequently fired a barrage of 59 cruise missiles at Shayrat air base in Syria to punish Damascus, despite its denials of responsibility.
Sadr, who led a militia that fought the US occupation of Iraq, also condemned the American missile strike, urging all foreign parties involved in the Syria conflict to withdraw.
"I have not only called for the resignation of Bashar, but I had already called for Abedrabbo and the ruler of Bahrain to step down because they are still oppressing their people," Sadr said in his statement.
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