Takatsuki & Nitori Wandering Son is the definitive anime about transgender children. While not focused solely on "ladyboys," it features a male-to-female protagonist and is the most realistic depiction of gender dysphoria in the medium.
The classic. Haruhi Fujioka is a scholarship student at an elite academy who dresses as a boy to work in the Host Club. While not a "ladyboy" herself (she’s more of a tomboy/androgynous icon), the series is packed with gender-bending play and a beloved cross-dressing character, Renge. It’s the gateway anime for this whole subject. ladyboy anime