Anti-sickling activity of novel analogue of 3,4 dihydroxy cinnamic acid using in-silico studies and synthesis by inhibiting polymerization of hemoglobin

In India and in various regions of Central Asia and Africa, one of the most prevalent deadly and painful hematological disorders is sickle cell anemia, which is a major concern because of less social awareness and no curative treatment available. Disease-carrying individuals have shorter life spans with high morbidity and mortality. To treat and manage the root cause of this disease, novel drug discovery is a necessity. To identify best analogue of proposed derivative of 3,4 di-hydroxycinnamic acid which inhibits hemoglobin polymerization and useful for treatment of Sickle cell Disease. A series of analogues were created to determine docking scores of compounds utilizing Auto-Dock Vina software, and later redocking was done by other software to validate the best docked compound. Comparative analysis of binding energies and ADMET profiles of compounds with approved drugs was done to identify the best candidate, and to check stability, in silico simulation studies were performed to determine which would be the most effective, stable, and least hazardous compound. These were then synthesized and biologically assessed. We assessed that among the 14 analogues, the C-1 analogue exhibited superior binding affinities after using three software programs for molecular docking analysis. Additionally, MD simulation studies predicted better stability, as most residues in the RMSF were below 4.0 Å, with one base peak at the start and a larger peak observed at 2.0 Å. The radius of gyration remained within the range of 5.2 to 6.0 complex throughout the time period of the 100 ns simulation. Molecular dynamic simulation analysis shows that the complex remains stable all through the action, and the least toxic, better pharmacokinetic profile was assayed through ADMET software and later synthesized in the wet lab, and the newly synthesized compound was confirmed through IR, NMR, and mass data analysis, which also showed better anti-polymerizing activity confirmed through sickle cell-carrying human blood and normal human blood studies; the antioxidant activity of the compound is also confirmed by assay. The best activity was shown by Compound C-1 by inhibiting the polymerization of RBCs and having antioxidant activity with a good pharmacokinetic profile than already existing USFDA drugs. Only symptomatic treatment management of drugs is available still. In this study, a novel analogue of 3,4-dihydroxycinnamic acid is showing better results as an anti-sickling activity by inhibiting polymerization of haemoglobin than the already existing approved drug for management of sickle cell anaemia, as it is one of the prevalent genetic hemoglobinopathies. Results are validated through molecular docking studies, molecular dynamic studies, and synthesis of the best compound. Further animal toxicity studies are reported, and anti-sickling activity is shown through human blood studies.