Can we have more details?
Specifically:
What is the house now?
What are the surrounding houses like?
Have any of them extended?
Will some of your options make the house top heavy?
And by 3rd bed with small ensuite do you mean the "master" bedroom?
Do you have a downstairs toilet?
I think 3 or I'm leaning towards 4. But that's me persionally because I've always had a smallish bathroom and to be honest a big one just needs more cleaning. I would however love a little study more than a bit extra bathroom space.
I don't really do much or move about that much in a bathroom! And the smaller it is the less room for slippery wet little naughty children to evade you in. A small room and you've got them penned in between the bath and the wall with you blocking the door

One gets done and then sits singing on the toilet seat. Works fine.
So option 4 for me, but I'm going off my personal preference.