In Bombay Velvet, Kaizad Khambatta is married, but uses his wife for honey-trapping his enemies, and seems to prefer the company of other men and seems very jealous of Johnny's romance with Rosie.
Prince Edward in Braveheart, a foppish, homosexual weakling who cares more about new clothes and playing with his boy toy than conquering Scotland. (His Real Life analogue, Edward II, actually was gay, but not an example: besides the problems with calling real people villains, the real Edward II was interested in traditionally manly, outdoorsy pastimes like sports and, unusually given his station in life, manual labor.) The main villain, Longshanks, is a manly man.
"Sally Can't Dance" in Con Air. How effeminate is he? Well, his nickname is "Sally," he refers to men as a separate gender, he wears dresses, and in his final confrontation with the hero, Nicolas Cage's character winds up for a punch, visibly reconsiders, and slaps Sally in the face instead.
Sebastian Valmont in Cruel Intentions. Not gay, certainly, but soft-voiced and whiny. The clincher? His stepsister promises him that if he succeeds in an evil task she's giving him, she'll fuck him.
Blofeld from the same movie got quite the upgrade in fabulousness, as well. He even got to dabble in Creepy Crossdressing.
The Scorpio Killer from Dirty Harry. The contrast is made more blatant by the fact that Harry Callahan is played by quintessential "man's man" Clint Eastwood.
From the 70s film The Eiger Sanction (based on the book), there's a fellow called Miles Mellough, whose sole purpose in the movie is to embody this trope. He has a lap dog named 'Faggot', wears flamboyant clothing and makes off-colour jokes about rape in a very sissy voice. At no time is he a credible threat (we're supposed to believe he was a crack commando?!) and his death is devoid of any interesting fight: Eastwood leaves him to die in the desert midway through the film.
Jean-Baptiste Emanuel Zorg from The Fifth Element had some of these traits, specifically the delicacy and high fashion. Remember his perfectly appointed office? Dallas, on the other hand, is an ex-soldier played by Bruce Willis.
The villain in the Chow Yun-fat film Full Contact is openly gay and wants to fuck Chow almost as much as he wants to kill him.
Park Chang-yi in The Good, the Bad, the Weird is a murderous, psychopathic prettyboy with anachronistically fabulous clothes and grooming. Overlaps with Agent Peacock, as he's the most feared assassin in Manchuria and every bit as deadly as he is pretty.
Percy in The Green Mile. Fittingly, the casts' Southern accents make it sound a lot like "pussy."
One of the complaints often levied against the Harry Potter films is that Voldemort often comes across as this.
Herod in the original movie production of Jesus Christ Superstar. He lounges in a swimming pool with two attractive women and two male servants, sings in a higher register than the other male characters and does a very camp dance during his song.
Valentine from Kingsman: The Secret Service has a lisp, dresses fairly casually (and when he DOES go formal, is very foppish), and is sickened by acts of murder. Direct murder, that is. He's fine with having people kill each other.
The Maltese Falcon pits macho Anti-Hero Sam Spade up against three Sissy Villains: Joel Cairo, Kasper Gutman, and Wilmer. (Only The Hays Code prevented the film from out-and-out showing that Cairo and Wilmer are the Gayngsters they are in the novel.)
In Abel Gance's biopic of Napoleon, Gance himself steals the show as Louis Antoine de Saint-Just. Foppish and effete in his dangling earrings, and described as the most feared man in the French Revolution other authors have called him the Terror's "Angel of Death".
The alien commander Eros. In the words of the Rifftrax guys: "He makes Carson Kressley look butch!" The actor's name was Dudley Manlove. Yes, that was his real name. Poor guy.
Eros' superior, however, out-swishes him by a mile. He was played by a guy with the ordinary name (and not-so-ordinary nickname) of John "Bunny" Breckinridge, but his lifestyle topped having the surname "Manlove" by ten miles.
Watson from Razor Sharpe, although, given his unfathomable accent, it's anyone's guess whether this is intentional or just the result of dodgy acting.
Repo! The Genetic Opera has Parvi Largo as a minor villain. He is foppish, flamboyant and painfully vain; he dresses in replica vintage Italian suits, and speaks with an outrageous Italian accent for no good reason. He's also a serial rapist who wears the face of a beautiful woman over his own.
Archibald Cunningham from Rob Roy, a downright fop who murders people to buy fancy new clothes. He affects a comically mincing facade, but drops the act behind closed doors and when he's angry. When talking to his mistress he actually mentions putting on a "lithp" as part of his highborn act. True to the way this trope is usually played, The Hero is the classically manly Rob Roy, though unusually, Cunningham is a vastly superior fighter with a fair amount of Villainous Valour.
Jean Girard from Talladega Nights: The Ballad of Ricky Bobby is a comedy version of this trope. He even talks about his husband. And he raises horses - who are also gay. However, subverted in that he's the best racer on the circuit for a time - and he puts Ricky in a humiliating arm lock in order to force him to admit that French crepes are delicious.
Willard Gates, the villain with an aversion to being exposed to violence, in This Gun for Hire.
Darren "Wall Street" Bettencourt of The Transporter spends his first appearance trying to feel Frank up, while talking about how much "I like him." He's got the limp wrists, the Large Ham gestures, the pretty boy look, and the mincing. And then he reveals that he's actually The Dragon and a fairly nasty Blood Knight who wants to fight Frank to the death. Anyone else see that coming?
Subverted by Adrian "Ozymandias" Veidt in Watchmen. The screenwriter and director were convinced he was gay, the actor who played him suggested he was probably at least bisexual in an interview, and there's an Easter Egg in a shot of his computer desktop that may or may not have a gay Porn Stash. He's also extremely well-dressed, to the point of garishness in some scenes, and has a fabulous androgynous haircut bordering on Power Hair. He's also a ruthlessly efficient martial artist, a hard-nosed businessman, and has terse, rather dry and dour speech patterns and body language. He might not be quite Straight Gay, but there is nothing mincing or effeminate about him in the slightest.
Mr. Ho, The Dragon from Way of the Dragon, is quite effeminate, especially when he feels Tang Lung's stomach and says, "Ohhhh, what rippling muscles!"