Chemical profiling and in silico analysis of phytocompounds extracted from milky-latex sap from giant milkweed Calotropis gigantea (Linn.) targeting nitric oxide synthase (iNOS) and cyclooxygenase-2 (COX-2) proteins

The giant milkweed, or Calotropis gigantea , is a well-known plant that produces milky substances used in alternative and traditional medicine to treat various illnesses. The current study’s objectives were to extract phytochemicals using solvent dependence and assess the anti-oxidant and anti-inflammatory properties of solvent-based latex extracts (acetone, methanol, ethanol, iso-propyl alcohol, and iso-amyl alcohol) extracted from the rapidly expanding Calotropis gigantea. This study targets nitric oxide synthase (iNOS) and cyclooxygenase−2, (COX-2) two proteins linked to inflammatory response, by identifying certain phytoconstituents from Calotropis gigantea milky sap using both in vitro testing and in silico-dependent prediction. Phytochemical testing of phenolics, flavonoids, and tannins as well as antioxidant and anti-inflammatory assays were conducted using standard methods. A gas chromatography-flame ionisation detector (GC-FID) was used to compare the bioactive chemicals in latex extracts. The GC-FID results showed that the ethanol fraction had a maximum of 29 phytocompounds, whereas the isoamyl alcohol extract had 23 phytocompounds. Phenolics, flavonoids, and tannins were found by phytochemical analysis, however their content varied depending on the solvent. Among all solvents, CEE (Calotropis ethanol extract ) depicted high oxidant potential having IC 50 values (µg/ml) in following order: NO 2 Scavenging Activity: 14, Hydroxyl Radical Scavenging Activity: 15, Iron Chelating Activity: 117. Notably all extracts displayed substantial anti-inflammatory activities. However, CAE (calotropis acetone extract) depicted maximum inhibition potential as evidenced from BSA denaturation fluorescent data. For instance: after the addition CAE to BSA fluorescence intensity went to 0.1 ×10 4 INT units from 3.6 ×10 4 INT units. Docking of three major phytocompounds [Phytol (43.61%), Ethion (18.67%), and Diisobutyl Phthalate (11.68%)], (abundance wise) as identified by GC-FID in CAE extract, revealed high affinity against iNOS and COX-2 enzymes having vina scores ranging from −8.3 and − 7.5 kcal/mol, respectively. Based on in vitro and in silico data, it was concluded that Calotropis gigantea latex could be used as anti-inflammatory agents, however, warranting further research using in vivo and clinical approaches.