Spider-Man 3 Theory: We Already Know The Villains...
4. "The Sinister Six"
Let's not beat about the bush here, Osborn will need to employ some underhand techniques to double down on his Jameson-led campaign against Peter Parker. He needs some heavies, in other words.
So what if he turns to the villains who are still out there who have a vendetta against Peter Parker? We already know that Mac Gargan has vengeance on his mind, because of the Homecoming post-credits and it's not like you cast someone as impressive as Michael Mando and then don't use him. So that's Scorpion involved and since he was basically created by J Jonah Jameson in the comics in a quest to undo Spider-Man, there's even more reason to think he might play a part.
He wouldn't be alone either, because Peter has put other villains away who might have a reason to be unhappy - including the likes of Shocker and the Avengers bank robbers - and The Tinkerer would appear to still be at large. If they could bring a big enough group of them together, don't put it past Marvel having Osborn refer to them - in passing - as his "sinister six" as he sets them out on the path to destroy Spider-Man. He needs him removed from the picture and he won't have any doubts that it will take a huge effort.
And what if he ends up getting a major assist from one of the most famous Spider-Man villains ever...?