Christopher Daniel Barnes
Peter Parker / Spider-Man (voice)

Sara Ballantine
Mary Jane Watson (voice)

Jennifer Hale
Felicia Hardy (voice)

Linda Gary
Aunt May Parker (voice)

Gary Imhoff
Harry Osborn (voice)

Ed Asner
J. Jonah Jameson (voice)

Rodney Saulsberry
Joe "Robbie" Robertson (voice)

Roscoe Lee Browne
Kingpin (voice)

Joseph Campanella
Dr. Curt Connors (voice) / Dr. Curt Connors / Lizard (voice) / Lizard (voice)

Nick Jameson
Michael Morbius (voice) / Morbius (voice) / Richard Fisk (voice) / Dr. Groitzig ...

Majel Barrett
Anna Watson (voice)

Maxwell Caulfield
Alistair Smythe (voice) / Ultimate Slayer Smythe (voice)

Neil Ross
Norman Osborn (voice) / Norman Osborn / Green Goblin (voice) / Green Goblin (voi...

Liz Georges
Debra Whitman (voice)

Patrick Labyorteaux
Flash Thompson (voice)

Dawnn Lewis
Detective Terri Lee (voice)

David Warner
Landon (voice)

Efrem Zimbalist Jr.
Doctor Octopus (voice) / Otto Octavius / Doctor Octopus (voice)

Joan Lee
Madame Web (voice)

John Beard
Newscaster (voice) / News Reporter (voice) / News Announcer (voice)