The present invention relates to an amphiphilic triblock copolymer comprising a poly(2-vinylpyridine) block and a poly(alkylisocyanate) block and the preparation method thereof, and particularly relates to an amphiphilic triblock copolymer comprising a coil-shaped hydrophilic poly(2-vinylpyridine) block and a rod-shaped lipophilic poly(alkylisocyanate) block, having a controlled structure of coil-rod-coil or rod-coil-rod, and the preparation method thereof.