According to Charlie's manager, he died from leukemia Wednesday morning in a New York City hospital, reported TMZ.
Charlie was perhaps best-known for his appearances on Dave Chappelle's show.
The popular stand up comic co-wrote and appeared in some of his brother's starring films, including "Norbit" and "Vampire in Brooklyn".
Charlie entered the industry after working as a bodyguard for Eddie at the beginning of his career.
In 2005, he starred in "King's Ransom" as a gay ex-con, who is hired by King (Anderson) to kidnap him in a fake kidnapping.
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"We just lost one of the funniest most real brothers of all time . Charlie Murphy RIP," Chris tweeted.
Hart wrote, "Wow... This is crazy. All I can say is RIP. Thank you for not only being a friend but for believing in me when I was young in this comedy game. I'm lucky to have know you and I'm even luckier to be able to say that I was a friend."
Zendaya posted, "Rest In Peace to the legend that is Charlie Murphy, thank you for always bringing us joy and laughter."
Spike shared, "My Late Brother-The Very Funny Charlie Murphy. This Picture Is From JUNGLE FEVER, Charlie Was Also In MO' BETTER BLUES Too. Rest In Power."
Debra wrote, "RIP Charlie Murphy. A true talent. You will be miss!"
Charlie's wife, Tisha Taylor Murphy, died in 2009 from cervical cancer. They shared two children, and the comedian also had another child from a previous relationship.