Identification of novel therapeutic targets for ovarian hypofunction through hormonal pathway regulation.

Ovarian hypofunction (OH) is a multifactorial endocrine disorder characterized by impaired folliculogenesis, hypoestrogenism, and subfertility, affecting an estimated 10–12% of reproductive-aged women worldwide and up to 1% of women under 40 years of age. Current interventions, including hormone replacement therapy (HRT), gonadotropin stimulation, and assisted reproductive technologies (ART), remain largely symptomatic, carry safety concerns, and fail to restore intrinsic ovarian activity. To address this unmet need, we employed an integrative computational strategy combining systems-level biological insights with immunoinformatics approaches to identify and evaluate a multi-epitope peptide candidate targeting the follicle-stimulating hormone receptor (FSHR). Seven FSHR-derived peptides with high predicted immunogenic potential were identified, validated for non-allergenicity and non-toxicity, and fused with flexible linkers into a 62-amino-acid construct. Physicochemical analysis indicated stability (instability index 38.80), thermostability (aliphatic index 89.68), hydrophilicity (GRAVY −0.365), and favorable solubility (Protein-Sol 0.584 compared with an E. coli average of 0.45). Codon optimization improved the codon adaptation index from 0.64 to 0.90 and reduced GC content from 63.98% to 53.23%, suggesting high expression potential in E. coli. Molecular docking with TLR2 yielded a favorable docking score (−296), supported by extensive hydrogen bonding and hydrophobic interactions. Long-timescale molecular dynamics simulations (500 ns) supported structural stability, with RMSD plateauing at approximately 0.3 nm, a compact radius of gyration (2.52–2.55 nm), and a persistent hydrogen-bond network (1250–1400 bonds). In silico immune simulations suggested balanced humoral and cellular immune responses, characterized by early IgM production, class switching to IgG, IL-2–associated T-cell expansion, and immune memory formation. Overall, this computational study identifies a structurally stable and immunologically promising peptide candidate for ovarian hypofunction, providing a rational basis for future experimental validation.