Irritable Bowel Disease (IBD), encompassing Crohn's disease and ulcerative colitis, remains a challenging condition linked to TNFSF15-mediated inflammation of the gastrointestinal tract. The exact etiology of IBD remains unclear, but it is believed to result from a complex interplay of genetic, environmental, and immunological factors (Ananthakrishnan, 2015). One significant molecular player in the pathogenesis of IBD is Tumor Necrosis Factor Superfamily Member 15 (TNFSF15), a cytokine involved in the regulation of immune responses and inflammation. Elevated levels of TNFSF15 have been associated with increased inflammation and tissue damage in IBD patients, making it a critical target for therapeutic intervention (Chen et al., 2018). Peptide-based therapies have gained substantial attention in recent years due to their high specificity and low toxicity. Macrotermes bellicosus, a termite species, and Curcuma longa, commonly known as turmeric, have both been identified as sources of bioactive peptides with potential therapeutic properties. Peptides derived from Macrotermes bellicosus exhibit anti-inflammatory and immunomodulatory effects, while Curcuma longa is well-known for its anti-inflammatory, antioxidant, and antimicrobial activities (Fitzgerald et al., 2019, Hewlings and Kalman, 2017). The conjugation of peptides from these sources presents a promising strategy for enhancing their therapeutic efficacy and stability, particularly in the context of targeted interventions for IBD. TNFSF15, also known as TL1A, is a member of the tumor necrosis factor (TNF) ligand superfamily and plays a pivotal role in the regulation of immune responses. It functions primarily through its receptor death receptor 3 (DR3), activating multiple downstream signaling pathways, notably the NF-κB and MAPK pathways. These pathways are crucial in mediating inflammatory responses, promoting cytokine secretion, T-cell proliferation, and apoptosis regulation. In the context of Irritable Bowel Disease (IBD), TNFSF15 is particularly significant due to its involvement in the differentiation and activation of Th1 and Th17 cells, which are known contributors to chronic intestinal inflammation. Overexpression or dysregulation of TNFSF15 has been strongly associated with increased mucosal inflammation, elevated cytokine levels (such as IL-6, TNF-α, and IFN-γ), and epithelial barrier disruption, all hallmarks of IBD pathophysiology. Thus, targeting TNFSF15 offers a promising strategy for modulating immune imbalance in IBD (Bamias et al., 2013).

An in-silico approach, which involves computational methods for simulating and analyzing biological systems, offers a powerful tool for designing and optimizing peptide conjugates. Computational techniques can predict the binding affinity of peptides to their targets, assess their stability, and simulate their interactions within the biological environment. By employing molecular docking and dynamics simulations, researchers can identify optimal conjugation strategies and refine peptide structures for improved efficacy against TNFSF15 (Morris and Lim-Wilby, 2008). This approach not only accelerates the drug development process but also reduces the need for extensive in-vitro and in-vivo testing. Gene expression analysis further complements the in-silico approach by providing insights into the biological effects of peptide conjugates on TNFSF15 expression and related inflammatory pathways. Techniques such as quantitative PCR (qPCR) and RNA sequencing (RNA-seq) allow for the quantification and characterization of gene expression changes in response to peptide treatment. This dual approach, combining computational modeling with gene expression analysis, enables a comprehensive evaluation of the therapeutic potential of Macrotermes bellicosus and Curcuma longa peptide conjugates for targeted intervention in IBD (Schena et al., 1995, Wang et al., 2009). The integration of these methodologies represents a novel and innovative strategy for developing effective treatments for IBD, addressing both the molecular mechanisms underlying the disease and the practical aspects of drug design and delivery.

The aim of this study is to develop a targeted intervention for Irritable Bowel Diseases (IBD) by leveraging peptides derived from Macrotermes bellicosus and Curcuma longa, focusing on their potential to modulate the expression and activity of Tumor Necrosis Factor Superfamily Member 15 (TNFSF15). The study commenced with the analysis of TNFSF15 gene expression using the GTEx Portal, followed by functional enrichment and interaction analysis through g and STRING databases, respectively. Protein sequences from M. bellicosus and C. longa were retrieved from UniProt, and their gastrointestinal stability and bioactivity were assessed using BIOPEP-UWM and Peptide Ranker. Allergenicity and antigenicity of the selected peptides were evaluated using Allertopv.20 and the Scratch Protein Predictor tool. The peptides were conjugated with a 50S ribosomal protein L7/L12 adjuvant using flexible linkers and a 6X Histidine tag for enhanced efficacy and stability. Molecular modeling and structural validation of the conjugated peptides were performed using trROSETTA and PROCHECK, while the TNFSF15 protein structure was retrieved from AlphaFold. The interactions between TNFSF15 and the conjugated peptides were explored through molecular docking with ClusPro and further validated by molecular dynamics simulations using the MD Web Server. Lastly, immune simulations were conducted using the C-ImmSim SERVER to predict the immunogenicity and antigenicity of the conjugated peptides, providing a comprehensive approach to developing an effective therapeutic strategy for IBD.