## 2 Results
### Inhibition of Nrf2 Recruitment by the Keep1-Ligand Complex  
To explore how Nrf2 interacts with Keep1 in the presence of apigenin, luteolin, omaveloxolone, and the Cpd16 compound, peptide-receptor docking simulations were performed. The binding affinity was measured using total energy. The Nrf2 structure was sourced from a crystal complex with Keep1 (PDB ID: 2 FLU) [27]. We first validated the DockThor (a receptor\(-\)ligand docking program) method to set up the parameters for our simulations [28]. This validation involved re-docking, during which we observed Root Mean Square Deviation (RMSD) and analyzed active site interactions to pinpoint possible binding sites for Nrf2. The results were fine-tuned to ensure precision, providing a clearer picture of how Nrf2 binds to Keep1. This detailed analysis is essential for developing effective therapeutic interventions targeting these interactions. The RMSD value from the re-docking results was 1.7652 A (Table 3), which validates the parameters used in the peptide\(-\)receptor docking simulations. This low RMSD value reflects the accuracy of the docking approach, indicating that the predicted binding conformation closely resembles the experimentally determined structure. The consistent identification of key interacting residues, such as ARG334, SER363, ARG415, and TYR572, further supports the reliability of the docking results. These residues play a crucial role in stabilizing the ligand\(-\) receptor complex through hydrogen bonding and electrostatic interactions, enhancing the precision of the docking process. Based on Table 4, the 4 IQK-apigenin complex shows the lowest total energy of 993.266 kJ/mol in its interaction with Nrf2, compared to luteolin (993.573 kJ/mol), omaveloxolone (993.991 kJ/mol), and Cpd16 (993.960 kJ/mol). Although apigenin's binding affinity (\(-\)1.162 kJ/mol) is lower than the others, the lower total energy indicates better interaction stability, supported by an electrostatic energy of \(-\)2.430 kJ/mol. This suggests that apigenin could be a more effective candidate for inhibiting the interaction between Nrf2 and Keep1, thus having greater potential in therapeutically influencing the Nrf2 signaling pathway. Additionally, the lower total energy suggests that the interaction between apigenin and Nrf2 is more energetically favorable, which could contribute to a more stable complex formation. This stability could enhance the inhibitory effect of apigenin on Nrf2's interaction with Keep1, making it a promising candidate for further therapeutic investigation. This calculation helps us understand how stable each complex is after peptide-receptor docking. Our results show that the 4 IQK-apigenin complex is more effective at blocking Nrf2 binding compared to the other 4 IQK-ligand complexes. This unique property of \begin{table}
\begin{tabular}{c c} \hline \hline
**Amino Acid Residue** & **RMSD and Docking Conformation** \\ \hline ARG334 & \\ SER363 & \\ ARG380 & \\ ASN382 & \\ ARG415 & \\ ARG483 & \\ SER508 & \\ GLN530 & \\ SER555 & \\ TYR572 & \\ SER602 & 1.7652 Å \\ \hline \hline \end{tabular}
\end{table} 
Table 3: Findings from the validation process of the peptide\(-\)receptor docking technique. appienin seems to stem from its strong interaction capabilities, which inhibit Nrf2 in cellular environments and reduce corepressor peptide binding. Specifically, Nrf2 interacts with key regions on Keap1, such as loop D-A and B-C (shown with a red box for emphasis), which are crucial for regulating Nrf2 degradation and maintaining the stability of the protein\(-\)ligand complex (Figure 2) [27]. The enhanced binding affinity of apigenin likely disrupts these interactions more effectively than other ligands, leading to a more pronounced inhibition of Nrf2 activity.