Artificial recapitulation of hierarchically porous films gained great interest due to their versatile functionalities and applications. However, the development of novel eco-friendly and nontoxic biopolymer-based porous films is still limited by the time-consuming fabrication processes and toxic organic reagents involved. Here, we reported a novel approach to rapidly (within 5 s) fabricate biopolymer-based hierarchically porous films via inducing the laterally occurring interfacial self-assembly of prolamins at the air-liquid interface during an antisolvent dripping procedure. The as-prepared films exhibited a hierarchically porous microstructure (with sizes of about 500 nm to 5 μm) with location-graded and Janus features. The formation mechanism involved the solvent gradient controlled self-assembly of prolamin into an anisotropic defect structure in longitudinal and lateral directions. Accordingly, the macroscopic morphologies together with the porosity and pore size could be precisely tuned by solvents and operating parameters in a convenient way. Furthermore, alcohol-soluble but water-insoluble bioactive compounds could be incorporated simultaneously via a one-step loading procedure, which endowed films with large loading efficiency and sustained release features suitable for controlled release applications. The effect of the curcumin-loaded porous film on skin wound healing, as one of the potential applications of this novel material, was then investigated in vivo in a full-thickness wound model, wherein satisfying wound healing effects were achieved through multitarget and multipathway mechanisms. This pioneering work offers a novel strategy for the rapid architecture of biopolymer-based hierarchically porous film with versatile application potentials.