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The series was a spin-off from Wacky Races and featured race driver Penelope Pitstop (voiced by Janet Waldo). She wore a bright pink auto-racing outfit. Pitstop was constantly being chased by the villian Hooded Claw, aka Sylvester Sneekly (voiced by Paul Lynde) who wanted to get rid of her in order to get her inheritance. Penelope was often rescued from peril by the Anthill Mob, a group of diminutive gangsters led by Clyde, a charicature of Edward G. Robinson in Little Caesar.
The title was a reference to a silent movie era melodrama cliffhanger movie serial, The Perils of Pauline. While the earlier Wacky Races series the heroine came from looked to be set contemporarily in the 1960s, most of the automobiles and clothing of The Perils of Penelope Pitstop gave it the look of being set in the 1920s or 1930s.
Other voice artists in the series included Mel Blanc and Garry Owens.