Cyclosporine (CsA), a potent immunosuppressive chemotherapeutic medication, treats numerous cancers, particularly malignant carcinoma, acute leukemia, and triple-negative breast cancer (TNBC). A specified polymeric nanoformulation (N.F.) based drug delivery technique with ligand alteration at the surface was developed to improve active moiety delivery at the intended area and boost the efficacy of prolonged treatment. We produced and characterized N.F. of encapsulated Cyclosporine in thiolated chitosan (T.C.) with the outermost coating of hyaluronic acid (H.A.). Research in-silico verified that H.A. binds to the receptor CD44 at docking locations A and B in triple-negative breast cancer cells. When a pharmaceutical substance interacts with a polymeric compound, zeta examination reveals a particle size of 192nm, a PDI of 0.433, and zeta potential of 38.9 mV. FTIR and Raman's investigations also support the existence of hydrophobic groups, porous surfaces, and clumping characteristics. While XRD verified its crystallographic nature, which renders N.F. particularly helpful in localized drug delivery systems (DDS), SEM and TEM revealed circular nanoparticles with sleek exteriors. DSC demonstrated that N.F. was stable at high temperature. The NF showed 85% drug encapsulation, and a kinetics investigation of drug release revealed that the N.F. obeyed the Higuchi model for dispersion at low pH. In contrast to typical CsA's immediate release in under 12 hours, the in-vitro investigation demonstrated prolonged continuous dissolution at pH 7.4 and 6.8 for a maximum of 72 hours. When compared to raw Cyclosporine, the in-vitro tumor prevention properties of the ThC-HA encapsulated with Cyclosporine were tested using an MTT test on normal breast epithelial cells and triple-negative breast cancer cells. It showed the synthesized N.F.'s robust cytotoxic potential at reduced concentrations and its effectiveness for normal cells. These characteristics improve the long-term viability, effectiveness, and active targeting of prepared novel N.F.s as an effective pharmaceutical component and potent therapeutic moiety against cancer.