Introduction: Itch, an uncomfortable sensation leading to the urge to scratch, is often inadequately managed by conventional antihistamine drugs, which can be ineffective in certain pruritic conditions and cause undesirable side effects. Therefore, there is a need to identify new pharmacologically potent antipruritic compounds with fewer side effects. A synthetic curcuminoid analogue, 2,6-bis-(4-hydroxy-3-methoxybenzylidene)-cyclohexanone (BHMC), a derivative of curcumin - a bioactive compound found in turmeric - has demonstrated various pharmacological ac-tivities. Previous studies have shown that BHMC possesses antinociceptive and anti-inflammatory properties. This study aimed to investigate the antipruritic effects of BHMC in mice models of induced pruritus. Materials and Meth-ods: The pruritus in mice was induced using compound 48/80, substance P, histamine, and serotonin to establish an itch-induced mouse model. BHMC was administered intraperitoneally (i.p.) at doses of 0.1, 0.3, and 1.0 mg/kg. Results: BHMC significantly reduced pruriceptive responses in all models tested and notably inhibited compound 48/80 and substance P-induced mast cell degranulation in skin tissues. Conclusions: These findings suggest that BHMC inhibits pruriceptive responses in mice, likely through the inhibition of mast cell degranulation and/or direct antagonism of peripheral histamine and serotonin receptors. This may warrant further exploration of the antipruritic effect of BHMC in clinical trials for the betterment of animal and human health.

Introduction: Epithelial-mesenchymal transition (EMT) is a process of epithelial transformation into mesenchymal cells. It is also a process that contributes to the progression of fibrosis and cancer metastasis. Transforming growth factor-beta (TGF-β), as a potent inducer of EMT, has therefore became a potential therapeutic target. However, clinical developments of TGF-β inhibitors have been un-successful due to safety risks. Hence, drug repurposing of existing safe-to-use drugs could over-come this issue. Methods: In this study, the TGF-β receptor type 1 (ALK5) was selected as the target protein. Molecular docking was performed using known ALK5 inhibitors as positive controls. Clinical drugs with similar binding affinity and amino acid interaction were selected for in vitro experimental validation. Results: ALK5 inhibitor demonstrated binding affinities ranging from -11.2 to -9.5 kcal/mol. Analysis of amino acid interaction revealed that Val219, Ala230, Lys232, and Leu340 amino acid residues are crucial for binding. Subsequent screening of clinically approved drugs against ALK5 showed top five potential drugs (ergotamine, telmisartan, saquinavir, indinavir, and nelfinavir). The selected drugs were tested in TGF-β1-induced normal human bronchial epithelial cell line, BEAS-2B. Western blot analysis showed that the drugs did not exhibit inhibitory effects on the downregulation of epithelial proteins (E-cadherin) and upregulation of mesenchymal proteins (vimentin and α-smooth muscle actin). Conclusion: Based on these experimental outcome, it is postulated that the results from molecular docking were false positives. The tested drugs in this study could serve as negative controls in future screening against ALK5 protein.