Brand previously denied two counts of rape, one count of indecent assault and two counts of sexual assault in relation to alleged offences between 1999 and 2005, involving four women.
At Southwark Crown Court on Tuesday, the 50-year-old pleaded not guilty to one further count of rape and one of sexual assault, which are both alleged to have taken place in 2009.
Wearing a leopard print shirt with a number of buttons undone and holding a white hat, Brand spoke to confirm his name and his not guilty pleas from the dock.
The two latest charges relate to two further women and are alleged to have taken place in London.
A court previously heard Brand is accused of raping a woman in a hotel room while she attended a Labour Party conference in Bournemouth, and grabbing a TV worker's breasts and orally raping her after dragging her into a male toilet.
He is also alleged to have grabbed a radio station worker's face, pushing her against a wall and kissing her before groping her breasts and buttocks.
The other charge of the original five alleges the actor indecently assaulted another woman after grabbing her forearm and attempting to drag her into a male toilet.
Brand, who carried a copy of the Bible into the court building before his hearing, is due to face trial in June.
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