Nonradiative recombination losses occurring at the interface pose a significant obstacle to achieve high-efficiency perovskite solar cells (PSCs), particularly in inverted PSCs. Passivating surface defects using molecules with different functional groups represents one of the key strategies for enhancing PSCs efficiency. However, a lack of insight into the passivation orientation of molecules on the surface is a challenge for rational molecular design. In this study, aminothiol hydrochlorides with different alkyl chains but identical electron-donating (-SH) and electron-withdrawing (-NH3+) groups were employed to investigate the interplay between molecular structure, orientation, and interaction on perovskite surface. The 2-Aminoethane-1-thiol hydrochloride with shorter alkyl chains exhibited a preference of parallel orientations, which facilitating stronger interactions with the surface defects through strong coordination and hydrogen bonding. The resultant perovskite films following defect passivation demonstrate reduced ion migration, inhibition of nonradiative recombination, and more n-type characteristics for efficient electron transfer. Consequently, an impressive power conversion efficiency of 25% was achieved, maintaining 95% of its initial efficiency after 500 hours of continuous maximum power point tracking.