AIM OF THE STUDY: This study explores the anti-inflammatory effect of TPTQ in silico, in vitro, and in vivo.
MATERIALS AND METHODS: In silico testing used the Gnina application, opened via Google Colab. The TPTQ structure was docked with the nuclear factor kappa B (NF-ĸB) protein (PDB: 2RAM). In vitro testing began with testing the cytotoxicity of TPTQ against Raw 264.7 cells, using the 3-[4,5-dimethylthiazol-2-yl]-2,5 diphenyl tetrazolium bromide (MTT) method. A phagocytic activity test was carried out using the neutral red uptake method, and interleukin-6 (IL-6) and tumor necrosis factor alpha (TNF-α) secretion tests were carried out using the enzyme-linked immunosorbent assay (ELISA) method. In vivo, tests were carried out on mice by determining cluster of differentiation 8+ (CD8+), natural killer cell (NK cell), and IL-6 parameters, using the ELISA method.
RESULTS: TPTQ has a lower binding energy than the native ligand and occupies the same active site as the native ligand. TPTQ decreased the phagocytosis index and secretion of IL-6 and TNF-α experimentally in vitro. TPTQ showed significant downregulation of CD8+ and slightly decreased NK cells and IL-6 secretion in vivo.
CONCLUSION: The potent inhibitory effect of TPTQ on the immune response suggests that TPTQ can be developed as an anti-inflammatory agent, especially in the treatment of Covid-19.
AIM OF THE STUDY: This study aims to evaluate newly derived compounds from Garcinia cowa Roxb., focusing on their ADMET profiles (Absorption, Distribution, Metabolism, Excretion, and Toxicity) and anti-inflammatory bioactivity. The assessment will be carried out using a combination of in silico and in vitro experiments to determine their pharmacological potential as anti-inflammatory agents.
MATERIALS AND METHODS: Isolation of compounds from Garcinia cowa Roxb. was carried out using column chromatography, purified with radial chromatography, and recycling HPLC. The compounds' structures were evaluated for their ADMET profiles and anti-inflammatory bioactivity using the NF-ĸB protein (PDB Code: 2RAM) as the target. The in vitro experiment was conducted using Raw 264.7 macrophages cell to assess cytotoxicity, phagocytic activity, IL-6, and TNF-α secretion. The determination of the anti-inflammatory mechanism is carried out by testing the activity of NF-ĸB and IKB-α using the western blot method.
RESULTS: We successfully analyzed the structure of a new compound from the bark of Garcinia cowa Roxb., named Garciacowanin (NC). In silico analysis suggests that the drug shows promising absorption potential, there are concerns related to its metabolism and toxicity that warrant further investigation during the development process and does not show mutagenic properties based on the negative AMES test results. There is a risk of hepatotoxicity (liver damage) and the drug can also interfere with the hERG II ion channel, which can cause side effects on the heart. The compound can affect the NF-ĸB protein, while in vitro studies have demonstrated its ability to suppress phagocytic activity, as well as the production of IL-6 and TNF-α. Western blot analysis suggests that NC's anti-inflammatory mechanism functions via the NF-ĸB signaling pathway.
CONCLUSION: NC has the potential to be developed as an anti-inflammatory agent with a mechanism of inhibiting the inflammatory response through the NF-ĸB signaling pathway.