## 4 Discussion  
Sulforaphane, found in abundance in cruciferous vegetables, is emerging as a versatile compound with numerous health benefits when incorporated into the diet. Rigorous research has revealed its promising potential to treat a variety of diseases. In particular, sulforaphane has demonstrated its ability to reverse leptin and insulin resistance, making it a valuable ally in the fight against diabetes and obesity [16; 58]. Beyond metabolic health, sulforaphane also shows promise in the treatment of neurodegenerative conditions such as Alzheimer's disease, Parkinson's disease and multiple sclerosis [59]. Moreover, the anticancer properties of sulforaphane have captured great attention in the scientific community [60]. Ongoing studies are delving into the intricate mechanisms through which sulforaphane may contribute to cancer prevention and treatment, shedding light on its potential as a natural compound with profound implications for human health. Most of the benefits attributed to sulforaphane arise from its activation of the NRF2-mediated antioxidant response [14; 60]. However, evidence suggests that this compound interacts with several target proteins, broadening the spectrum of its effects [61; 62]. The aim of this study was to identify other target proteins of sulforaphane to better understand its diverse beneficial effects. Taking advantage of target prediction servers with different algorithms, 11 potential sulforaphane-interacting proteins were identified. Evidence of interaction was found for four of them, showing inhibitory effects of sulforaphane on MIF tautomerase activity [50], reduction of CDK2 expression [51], modulation of the GSK3\(\beta\) signaling pathway [53; 54; 55], and inhibition of PARP1 activity [56]. After considering the remaining seven proteins for further analysis (CYP19A1, NAMPT, P2 RX7, CTSL, ADORA1, ADORA2A and MAOA), attention was given to the NAMPT protein due to its metabolic relevance and correlation with the functional enrichment analysis of overexpressed genes in sulforaphane-treated cells. The analysis highlighted its association with NADP+ 1-oxidoreductase activity, ketosterol monooxygenase activity and oxidative stress response. These processes are closely related to nicotinamide adenine dinucleotide (NAD+) metabolism [63]. NAMPT catalyzes the limiting step in the mammalian NAD+ salvage pathway, converting nicotinamide and 5\({}^{\prime}\)-phosphor-1-pyrophosphate to nicotinamide mononucleotide (NMN). Subsequently, NMN together with ATP is con verted to NAD+ by NMNAT1-3 (nicotinamide/nicotinic acid mononucleotide adenylyl-transferases) [64]. In addition to upregulating NAD+, NAMPT also elevates NADP+ and NADPH levels, stimulating oxidative stress systems such as glutathione and thioredoxin [65; 66]. This intricate interplay between NAD+ metabolism and cellular redox balance positions NAMPT as a crucial player in the regulation of oxidative stress [66]. The interaction of sulforaphane with NAMPT reveals a potential pathway for modulating cellular redox dynamics, warranting further investigation into its implications for health and disease. NAT functions as an allosteric activator of NAMPT, strategically positioning itself near the active site of the enzyme. The hydroxyl end of NAT aligns closely with the amide groups of NAM and NMN, resulting in a subtle adjustment of the enzyme's substrate binding affinity for NAM. Specifically, the oxygen atom of the hydroxyl of NAT forms a hydrogen bond with a water molecule and binds to the side chain of residue S275 of NAMPT. In addition, the interaction involves the central amide nitrogen of NAT and Y188 and H191, facilitated by another water molecule [44]. In the docking analysis, sulforaphane was observed to interact with NAMPT by forming hydrogen bonds. Specifically, the sulfinyl group of sulforapane established hydrogen bonds with the amino groups of residues K189 and R349 of NAMPT.