Curcumin is a hydrophobic polyphenol derived from turmeric with potent anti-oxidant, anti-microbial, anti-inflammatory and anti-carcinogenic effects. Curcumin is degraded into various derivatives under in vitro and in vivo conditions, and it appears that its degradation may be responsible for the pharmacological effects of curcumin. The primary risk factor for the cause of gastric cancer is Helicobacter pylori (H. pylori). A virulence factor vacuolating cytotoxic A (VacA) is secreted by H. pylori as a 88 kDa monomer (p88), which can be fragmented into a 33 kDa N-terminal domain (p33) and a 55 kDa C-terminal domain (p55). Recently it has been reported that curcumin oxidation is required to inhibit the activity of another major H.pylori toxin CagA. We performed molecular docking of curcumin and its oxidative derivatives with p33 and p55 domains of VacA. Further, we have examined the effect of the oxidation of curcumin on the vacuolation activity of VacA protein. We observed the binding of curcumin to the p55 domain of VacA at five different sites with moderate binding affinities. Curcumin did not bind to p33 domain of VacA. Remarkably, cyclobutyl cyclopentadione and dihydroxy cyclopentadione, which are oxidized products of curcumin, showed a higher binding affinity with VacA protein at all sites except one as compared to parent curcumin itself. However, cyclobutyl cyclopentadione showed a significant binding affinity for the active site 5 of the p55 protein. Active site five (312–422) of p55 domain of VacA plays a crucial role in VacA-mediated vacuole formation. Invitro experiments showed that curcumin inhibited the vacuolation activity of H. pylori in human gastric cell line AGS cells whereas acetyl and diacetyl curcumin, which cannot be oxidized, failed to inhibit the vacuolation in AGS cells after H. pylori infection. Here our data showed that oxidation is essential for the activity of curcumin in inhibiting the vacuolation activity of H. pylori. Synthesis of these oxidized curcumin derivatives could potentially provide new therapeutic drug molecules for inhibiting H. pylori-mediated pathogenesis.

Prakriti, the constitutionally determined phenotypic framework described in Ayurveda, is thought to capture biologically meaningful inter-individual variation in physiology, disease susceptibility and therapeutic response. Although prior Ayurgenomics studies have identified constitution-linked differences in gene expression, genetic variation and epigenetic state, the regulatory architecture underlying these phenotypes remains poorly understood. We applied a multi-layer systems biology framework to Prakriti-stratified bulk transcriptomic data. Weighted gene co-expression network analysis was used to identify Prakriti-associated modules and hub genes. Pathway activity was inferred using PROGENy and GSVA; transcription factor activity was estimated using VIPER; immune composition was assessed using ssGSEA, CIBERSORTx, and xCell; and immune-evasion markers were analysed using differential expression and clustering approaches. Dark kinase activity was profiled as a dedicated analytical layer, and cross-layer integration was performed using z-score-standardised feature matrices, correlation analysis, principal component analysis and linear interaction modelling. Eighteen co-expression modules were identified, of which three showed strong Prakriti associations: a Vata-linked module enriched for innate immune sensing and leukocyte trafficking, a Pitta-linked module enriched for chromatin regulation and ubiquitin-mediated proteolysis, and a Kapha-linked module enriched for RNA processing, DNA repair, adhesion and calcium-channel biology. PROGENy revealed a largely shared inflammatory and hypoxic macro-state across constitutions, indicating that broad canonical signalling did not itself explain group separation. Finer-resolution analyses uncovered marked divergence across regulatory layers. Vata combined innate immune activation with strong immunosuppressive features, including selective elevation of IDO2, ARG1, TGFB3, SIGLEC15 and CD274. Pitta showed mitochondrial and proteostatic signatures together with dominant PDCD1LG2 expression. Kapha displayed the most consistent GSVA elevation across immune-metabolic pathways and was enriched for M2 macrophages, NKT cells and B cells. Dark kinome profiling identified reciprocal constitutional patterns: ALPK3 and STK31 were enriched in Kapha, and BRSK1, WNK2, and STKLD1 were enriched in Vata. Cross-layer integration revealed a coherent TNFa/NFkB-to-AP1 regulatory bridge. It showed that dark kinase activity was significantly coupled to immune-evasion outputs, with checkpoint regulation displaying a Prakriti-dependent interaction that reversed in Pitta relative to Kapha. Prakriti-associated molecular divergence is not primarily encoded in isolated genes or broad pathway shifts, but in constitution-specific regulatory wiring across co-expression, transcription factor, immune, metabolic and dark kinase-linked layers. These findings move Ayurgenomics beyond descriptive stratification towards a systems-level mechanistic framework for constitutional biology and provide a basis for future validation in precision immunology and personalised medicine.