The play featured a number of then-prominent British actors including Michael Redgrave (as the Caterpillar), John Gielgud (as the Mock Turtle), and Peter Sellers (as the King of Hearts), as well as two of Miller's fellow cast members from Beyond the Fringe, Peter Cook and Alan Bennett as the Mad Hatter and the Mouse, respectively.

The title role was played by Anne-Marie Mallik, the 13-year-old daughter of a Surrey barrister, this being her only known acting performance. Wilfrid Brambell played the White Rabbit, Michael Gough and Wilfrid Lawson were the March Hare and the Dormouse, Alison Legatt was the Queen of Hearts, and Leo McKern did a drag turn as the Ugly Duchess. The journalist and broadcasting personality Malcolm Muggeridge was The Griffon. The play also featured a very young Eric Idle, several years before Monty Python brought him notice.