Simply put: it stinks, like bad. The room is hidden under the largest city in the land, only accessible by spelunking through vast distances of underground sewers that smell like decomposing waste. This smell-field ensures only the bravest and most dedicated adventurers ever travel to the room, and none who attempt it enjoy talking about it.
The room is built on the inside of a giant snail's shell, if you give it a special kind of lettuce the snail will retreat and clear access to the room.
The room is filled with a deadly gas. Either it is built so that it vents into the room naturally from deep under ground, or a diffuser is periodically refilled by whoever controls access to the room.
The McGuffin is the antidote to the gas, which you must obtain and administer before entering.
One way of getting a very localised danger like this would be to put it near radioactive ore deposits? You can only go in with a lead shield, and have to angle it correctly
Smoke and mirrors. Optical illusions. Puzzles. Riddles. Trap doors. Hidden sharp objects. Hidden blunt objects. Needing multiple people to complete something or turning multiple keys or keylike objects from other parts of the story perhaps?
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