"Sometimes in life, you think you found the one. But then one day you find out that she is just some b**ch," Timberlake said at the event.
It was perceived as a comment at Spears, who dated him for three years before parting ways in 2002, reports contactmusic.com.
Timberlake, who is now married to Jessica Biel, later took to Twitter and posted: "Ok. I see you. Wouldn't disrespect Anyone personally. Ever. Relax."