Osteoarthritis (OA) is an inflammatory disease that affects the cartilage and tissues around the joints, which results in excessive pain and stiffness. One of the most critical challenges for improving the therapeutic effect in osteoarthritis treatments is the current drug design utilizing functional polymers. Indeed, there is a need to design and develop novel therapeutic drugs for positive outcomes. In this view, glucosamine sulfate is a drug used to manage osteoarthritis (OA) because of its potential therapeutic effects on cartilage and ability to inhibit disease progression. This research aims to develop a Keratin/Chitosan/Glucosamine sulfate composite loaded functionalized multi-walled carbon nanotubes (MWCNT) as a potential carrier for the treatment of osteoarthritis. The nanocomposite was developed using various ratios of Keratin, Chitosan, Glucosamine sulfate, and MWCNT. Molecular docking analysis has been performed with (D-Glucosamine) and targeted proteins (PDB ID: 1HJV, 1ALU) to determine the binding affinity and interactions. Field Emission Scanning Electron Microscope (FESEM) study showed that the composite (KRT/CS/GLS) incorporated on the surface of functionalized multi-walled carbon nanotubes effectively. Fourier Transform Infrared spectroscopy (FTIR) analysis showed the presence of keratin, chitosan, and glucosamine sulfate in the nanocomposite and remained intact. X-ray diffraction (XRD) analysis indicated that the nature of the composite in MWCNT transformed from a crystalline to an amorphous state. Thermogravimetric (TGA) analysis revealed that the nanocomposite has a high thermal decomposition temperature of 420° C. The MTT assay results showed that 83% of cell viability has remained in RAW 264.7 cells at the maximum concentration (500μg/ml) of MWCNT-GLS/KRT/CS nanocomposite. Also, molecular docking results revealed the excellent binding affinity of D-Glucosamine to each protein structure (PDB ID: 1HJV and 1ALU).