The invention discloses a chip on film (COF) packaging substrate for a high-pixel camera module and a manufacturing method thereof. The COF packaging substrate comprises a soft core plate, and a first hard plate base material and a second hard plate base material which are adhered to the soft core plate, wherein the soft core plate is provided with a folding area; the first hard plate base material and the second hard plate base material are provided with windows corresponding to the folding area; the soft core plate comprises a soft plate base material, and a first copper foil layer and a second copper foil layer which are arranged on upper and lower sides of the soft core plate; a first covering film and a second covering film are adhered on the first copper foil layer and below the second copper foil layer; an image sensor chip is packaged in the first copper foil layer; the first copper foil layer is provided with a finger bonding area for COF packaging; and the first covering film and the first hard plate base material are hollowed corresponding to the finger bonding area. The manufacturing method comprises the steps of primary treatment, composite lamination and subsequent treatment. The COF packaging substrate is applicable to the high-pixel camera module and has the performance of dynamically and statically folding.