Komal Lakhani ¹ Rasmieh Hamid ^2,3* Sheetal Gupta ^4* Poojaben Prajapati ⁵ Ratna Prabha ⁶ Saumya Patel ⁵ Kirankumar P. Suthar ^7*

• ¹ Department of Biochemistry and Biotechnology, Junagadh Agricultural University, Junagarh, India
• ² Agricultural Research, Education and Extension Organization (AREEO), Tehran, Iran
• ³ Cotton Research Institute of Iran, Gorgan, Golestan, Iran
• ⁴ Navsari Agricultural University, Navsari, Gujarat, India
• ⁵ Gujarat University, Ahmedabad, Gujarat, India
• ⁶ Indian Council of Agricultural Research (ICAR), New Delhi, Delhi, India
• ⁷ Junagadh Agricultural University, Junagadh, Gujarat, India

Obesity, a prevalent global health concern, is characterised by excessive fat accumulation, which confers significant nutritional and health risks, including a shortened lifespan and diminished well-being. Central to the regulation of energy balance and food intake is the fat mass and obesity-associated (FTO) protein, which modulates the interplay between caloric consumption and energy expenditure. Given its pivotal role in obesity regulation, the identification of effective inhibitors targeting the FTO protein is imperative for developing therapeutic interventions. Currently available anti-obesity drugs are often plagued by undesirable side effects. In contrast, natural plant-derived bioactive compounds are gaining prominence in the pharmaceutical industry due to their efficacy and lower incidence of adverse effects. Little Millet, a traditional cereal known for its rich nutritional profile and high satiety index, was investigated in this study for its potential as an anti-obesity agent. Our research demonstrates that four bioactive compounds from Little Millet exhibit superior binding energies ranging from 7.22 to 8.83 kcal/mol, compared to the standard anti-obesity drug, orlistat, which has a binding energy of 5.96 kcal/mol. These compounds fulfilled all drug-like criteria, including the Lipinski, Ghose, Veber, Egan, and Muegge rules, and exhibited favourable profiles in terms of distribution, metabolism, and prolonged half-life without toxicity. Conversely, orlistat was associated with hepatotoxicity, a reduced half-life, and multiple violations of drug-likeness parameters, undermining its efficacy. Molecular dynamics simulations and Gibbs free energy assessments revealed that the four identified compounds maintain stable interactions with key residues in the FTO protein's active site. We propose further validation through extensive In vitro, In vivo, and clinical studies to ascertain the therapeutic potential of these compounds in combating obesity.