There could be a new dark wizard or witch who is specifically hunting down wizards who are hiding their true identities, including gender. This antagonist could have some personal vendetta against those who break the 'norm' in the wizarding world and see Harry as a prime target. They might use dark magic spells to detect and expose Harry's hidden gender.
Perhaps some traditional pure - blood families who have strict ideas about gender roles in the wizarding world. They might not approve of Harry's actions of hiding her gender and try to force her to conform to the traditional male identity. They could use their influence in the Ministry of Magic or within Hogwarts to make things difficult for Harry.
Maybe in this fanfic, Harry discovers a prophecy that is related to her gender. To avoid fulfilling a part of it that could lead to disaster, she hides her gender. In the process, she could form new alliances with female students who are more in tune with her new 'female identity'. She might also face challenges in classes like Defense Against the Dark Arts because the teaching methods are sometimes more tailored to male students in the original Hogwarts setup.
Another possible antagonist could be a power - hungry pure - blood family. They might see Harry as an impostor as the heir, believing that only a pure - blood should hold such a title. They would try to undermine Harry at every turn, spreading rumors and creating obstacles for him both in school and in the wizarding world.
New dark wizards or witches could emerge. They might target Harry's children, seeing them as a potential threat in the future due to their connection to Harry Potter. They could try to kidnap the children to use them as leverage against Harry or to turn them to the dark side.
Voldemort would still be a major antagonist. His quest for power and domination in the wizarding world would directly oppose the female Harry Potter, just as in the original series. His Death Eaters would also be a constant threat, causing chaos and harm.
It could be some of the pure - blood supremacists. They might see her as an abomination because of her connection to Hecate, who is not part of the traditional wizarding families. They would try to undermine her at every turn and make her life difficult at Hogwarts.
One possible main antagonist could be a dark wizard who is envious of Eros's powers or the relationship between Eros and Psyche. This dark wizard could be trying to disrupt their relationship and steal their magic for his own evil purposes. Maybe he was once rejected by a love, and seeing Eros and Psyche's love makes him want to destroy it.
Voldemort could still be the main antagonist. His goal of taking over the wizarding world wouldn't change just because of Harry's new heritage. In fact, he might see Harry as an even greater threat now that he has the potential of Yoruichi's powers. Another antagonist could be some of the pure - blood supremacists in the wizarding world who are afraid of the new kind of magic that Harry might bring in with his connection to Yoruichi.
Voldemort could still be a main antagonist. Even if Fem Harry has a new inheritance, Voldemort would see her as an even greater threat. He might try to steal her inheritance or stop her from using it to defeat him. His Death Eaters would also be involved, trying to carry out his evil plans against her.
The Death Eaters who still hold onto the old pure - blood supremacist views could be main antagonists. They might see the Heir of Arcturus Black as a threat to their ideology, especially if the Heir is trying to change the Black family's ways.
The main antagonists could be a secret organization that wants to use Harry's unique abilities for their own evil purposes. This organization could be made up of rogue wizards who have teamed up with evil spies from the Bond universe. They would see Harry as a key to achieving their goals of world domination, whether it's in the wizarding world or the Muggle world. Another possible antagonist could be a powerful dark wizard who has made a pact with a Bond - like super - villain to stop Harry from growing into a force that could oppose them.