## 4 Discussion  
This study provides a comprehensive computational pharmacology analysis of lycopene, revealing its multi-target interactions and potential pharmacodynamic actions in humans. Our findings highlight the importance of moving beyond conventional hub-centric network pharmacology approaches. While traditional analyses prioritize targets with extensive known interactions, this may overlook pharmacologically relevant proteins with high binding affinity but limited representation in curated databases. By classifying lycopene targets based on both binding affinity and network connectivity, we provide a more comprehensive framework that captures both structural relevance and biological context, particularly valuable for understudied natural compounds such as lycopene. The in-silico target prediction identified 94 candidate targets, with affinity values ranging from 57 nM to 14 mM, highlighting the broad spectrum of lycopene's molecular mechanisms [20; 30; 42; 43]. This is particularly not surprising considering the broad spectrum of health benefits reported following lycopene use [16; 21; 44; 45]. Notably, nuclear receptors constituted the largest proportion (37%) of target classes, consistent with previous findings that carotenoids often modulate gene expression via nuclear receptor pathways [46; 47; 48]. GPCRs represented  
Figure 3: Tissue-specific protein expression of the top five high-affinity targets (_MAP2K2_, _SCN2A_, _SCN3A_, _TOP2A_, and _TRIM24_) of lycopene in humans. Expression of _MAP2K2_, _SCN2A_, _SCN3A_, _TOP2A_, and _TRIM24_ was observed in 41, 5, 44, 29, and 30 different tissues. The Venn diagram shows the synergistic relationship between the tissues expressing the top five high-affinity targets of lycopene. _MAP2K2_, _SCN3A_, _TOP2A_, and _TRIM24_ were collectively expressed in 23 different tissues, while _SCN3A_ was the only lycopene target expressed in heart muscle. The numbers indicate the total number of tissues in which the targets is expressed. the second-largest category (18%), indicating lycopene's potential to modulate signalling at the membrane receptor level as well. Focusing on the top 15 targets of lycopene, six distinct target classes were identified, dominated again by nuclear receptors (40%) and GPCRs (26.7%), which is again consistent with prior reports indicating lycopene's ability to modulate transcriptional and signalling pathways involved in inflammation, metabolism, and cellular proliferation [6, 35, 42]. Among the 20 targets with sub-10 uM affinities, 6 targets (_MAP2K2_, _SCN2A_, _SLC6A5_, _SCN3A_, _TOP2A_, and _TRIM24_) exhibited high-affinity binding (<1 uM), suggesting these targets are most likely to be pharmacologically modulated at physiological concentrations of lycopene. While lycopene is known for its weak direct activity at most proteins due to its hydrophobic nature and limited aqueous solubility [49, 50], our concentration-affinity (CA) ratio analysis provided a critical pharmacodynamic context. While lycopene's bioavailability is limited under typical dietary intake, studies have shown that chronic supplementation or the use of bio-enhanced formulations can lead to its accumulation in tissue-specific niches such as the prostate, liver, and adrenal glands [51, 52]. Our analysis focused on identifying pharmacologically relevant protein targets and interaction thresholds, thereby providing a mechanistic basis for lycopene's potential therapeutic effects under conditions of sustained or optimized exposure. Only three targets (_MAP2K2_, _SCN2A_, and _SLC6A5_) showed CA ratios > 1 at reported human plasma concentrations (75-210 nM), suggesting these are the most pharmacologically viable targets under physiological conditions.