Miriam Margolyes called out John Cleese, Bill Oddie and Graham Chapman for being "vicious" to her when she was younger.
The Harry Potter actress, 80, said her experience with the three men - along with Humphrey Barclay, Tony Hendra and Tim Brooke-Taylor - was extremely negative.
During her youth, she spent three years at Cambridge University where she appeared in productions by the Cambridge Footlights.
It was the moment she first met John, 81 Bill, 80, and the late Graham.
Miriam revealed about her time starring in the Footlights revue of 1962: "The three years I spent at Cambridge gave me everything I have. That was a time when I was fully alive when I fully became myself.
"But I lost my smile a little when I performed in the Footlights.
"I didn't like the Footlights boys and they really didn't like me. They made that obvious."
She continued: "When I say 'they', I refer to a most distinguished group: John Cleese, Graham Chapman, Bill Oddie, Humphrey Barclay (later Head of Comedy at LWT), Tony Hendra and Tim Brooke-Taylor.
Miriam said in her memoir, shared by MailOnline, that she was the only girl in the production and she thought she was as "good as they were - and they didn't".
However, the actress revealed she "admired" Monty Python (created by and included John and Graham) and The Goodies (a comedy group with Bill).
Miriam added that the three men were geniuses but "not gentlemen".
She continued: “If you think about it, the Monty Python shows didn't feature funny women, only the occasional dolly bird.
"And I certainly wasn't that."
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The Miss Fisher's Murder Mysteries actress confessed she would "weep in her bedroom" at Newnham College because the men would treat her as if she were "invisible".
"The treatment I received from those Footlights boys was diminishing, pointed and vicious. On reflection, it is they who diminished themselves," Miriam continued.
“John Cleese, Bill Oddie and Graham Chapman were total s***s — and they have never apologised. The only one who did was the late Tim Brooke-Taylor.”
The Mirror have contacted Bill and John's reps for comment.
Broadcast by the BBC from 1969 to 1974, Monty Python's Flying Circus was created, written and performed by Graham, John, Terry Gilliam, Eric Idle, Terry Jones, and Michael Palin.
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