A novel mucoadhesive nanoparticle (NP) system combining rivastigmine and insulin was developed for intranasal delivery to enhance drug transport efficiency across the nasal mucosa. The NPs were synthesized using N,N,N-trimethyl chitosan (TMC) via ionic gelation, encapsulating both rivastigmine, a potent acetylcholinesterase inhibitor, and insulin. The NPs exhibited a zeta potential of 17.6 mV, a size of 187.00 nm, and a polydispersity index (PDI) of 0.29. In vitro and ex vivo studies demonstrated that the NPs significantly improved drug transport efficiency compared to free drug solutions. For rivastigmine, the transport efficiency was 73.3% for NPs versus 52% for solutions, and for insulin, it was 96.9% for NPs versus 21% for solutions. These results indicate that the combinational mucoadhesive NPs enhance nasal delivery efficiency, offering a promising strategy for simultaneous cerebral delivery of rivastigmine and insulin. The NPs also showed good biocompatibility with sheep nasal tissue, indicating their safety for nasal administration. This study highlights the potential of TMC-based NPs as a novel drug delivery system for treating Alzheimer's disease and other neurodegenerative conditions.A novel mucoadhesive nanoparticle (NP) system combining rivastigmine and insulin was developed for intranasal delivery to enhance drug transport efficiency across the nasal mucosa. The NPs were synthesized using N,N,N-trimethyl chitosan (TMC) via ionic gelation, encapsulating both rivastigmine, a potent acetylcholinesterase inhibitor, and insulin. The NPs exhibited a zeta potential of 17.6 mV, a size of 187.00 nm, and a polydispersity index (PDI) of 0.29. In vitro and ex vivo studies demonstrated that the NPs significantly improved drug transport efficiency compared to free drug solutions. For rivastigmine, the transport efficiency was 73.3% for NPs versus 52% for solutions, and for insulin, it was 96.9% for NPs versus 21% for solutions. These results indicate that the combinational mucoadhesive NPs enhance nasal delivery efficiency, offering a promising strategy for simultaneous cerebral delivery of rivastigmine and insulin. The NPs also showed good biocompatibility with sheep nasal tissue, indicating their safety for nasal administration. This study highlights the potential of TMC-based NPs as a novel drug delivery system for treating Alzheimer's disease and other neurodegenerative conditions.