During the game's introduction, Spider-Man and Quentin Beck battle for the Tablet of Law and Chaos. However, due to the damage it took from their duel, it bursts, releasing multiple pieces called stars that scatter across the multiverse. Quentin demands that Spider-Man retrieves the stars to re-power the fragment. Spider-Man does so, but apparently knows seemingly not that Quentin wants to take his revenge...
Sergei Kravenoff: Kravenoff appears in the game's first stage. He uses his star to become harder to hit, stronger, and faster. His alternative names are "Kraven the Hunter" and "Kraven".
Joseph Lorenzini: Referred to as the Hammerhead, Joseph Lorenzini appears as the game's second boss. He uses the star in his possession to grow guns on his arms and become stronger, which allows him to hold onto his weapons.
Hobgoblin: This being appears as the game's third boss. He gets his hands on one star from the Tablet of Law and Chaos to gain psychic powers, increasing his strength and allowing him to trick his opponents.
Maxwell Dillon: Referred to as Electro, Maxwell Dillon started off as your normal Homō Sapiēns, but gained electric powers. He uses his star to become huge, resulting in his transformation into the fourth boss. His life bar incorrectly calls him "Dam" rather than by either one of his names.
William "Flint Marko" Baker: Called the "Sandman", Flint is the fifth boss to be encountered. He is encountered in a mine commissioned by the Roxxon Energy Corporation. After getting his hands on the fifth star from the Tablet of Law and Chaos, Sandman gained increased Harenakinesis. His battle involves him using his powers to create duplicates of himself which assist in his attacks, but he can be defeated if you send a water barrel flying in his direction
Adrian Toomes: Referred to as the "Vulture", Toomes started off as a circus freak who bit off chicken heads. In battle, he uses his star to gain teleportation powers, but his primary weakness is light, making him similar to a vampire along with his voice.
Kron Stone: Kron, also called the "Scorpion", used to be completely human, but he was mutated, causing his body to become part scorpion. His star from the Tablet of Law and Chaos allows him to use egg-laying to create duplicates that assist in his conquest.
Sergeant Wade Winston Wilson: Also called "Deadpool", Wilson is the eighth boss in the entire game. In battle, he uses the star from the Tablet of Law and Chaos to create duplicates which assist in his attacks. Finish off his duplicates first and then take down the real deal, but avoid his bombs, preferably by climbing on the walls. Perhaps as a joke to his duplication powers, Deadpool is called "Deadpools" during his boss battle.
Cain Marko: Also called the "Juggernaut", Cain is fought within the big city. He gets his hands on his star during an attempt to escape the Silver Sable.
Normal Osborn: Referred to as The "Goblin", Norman haunts the tenth level. He and the Vulture are very similar in their backstory. His powers from his piece of the Tablet's power involve him getting claws on his hands and increased strength and speed.
Doctor Serena Patel: Referred to as "Doctor Octopus" (even though Doctor Otto Octavius had that title), Serena haunts the eleventh level. Her battle arena is in the Condensed Matter Reactor. She can be defeated if Spider-Man severs the cables and frees the assistants on his way. She uses biotechnology to attack Spider-Man
Carnage: This monster is seen in the game's penultimate stage. It destroys the Triskelion and possesses several SHIELD Agents. The Carnage is mutated with the last star from the Tablet of Law and Chaos. Its battle is unique, with the aberration concealing itself by constructing an abhorrent membrane that attacks with tentacles. The Spider-Slayers will use their weapons to break the membrane, but they also take damage if they become exposed to the monstrous being's duplicates. When Spider-Man exposes it, the Carnage goes all out to attack Spider-Man. The Carnage is destroyed when Spider-Man throws it under a collapsing roof.
Mysterio: This entity is the game's final boss. He was trapped within the legendary Tablet of Law and Chaos, but when Spider-Man retrieved the Tablet's stars for Quentin Beck, Mysterio was released from his bonds, possessing the mad stage magician in the process. Spider-Man has to fight him through four agonizing phases. Spider-Man eventually destroys Mysterio and sends Quentin to prison for trying to destroy the Universe.