During the 2-2 draw earlier this month, Little kicked out at Marc Joseph in an off-the-ball incident, and was given his marching orders by referee Mr. Miller.
They claimed he was innocent of any foul act, but their appeal has been turned down by the disciplinary panel.
A statement read: "It's very disappointing because Colin tells us he didn't do anything.
"The Hull player obviously made a meal out of something or nothing and it led to the red card being shown.
"Unfortunately, unless there is clear evidence to the contrary, the panel's hands are tied and they have to go with the referee's report."
As a result, he will be suspended for three league matches.