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 OA 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 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 (KRT/CS/GLS) composite loaded functionalized multi-walled carbon nanotubes (MWCNTs) as a potential carrier for the treatment of OA. The nanocomposite was developed using various ratios of KRT/CS/GLS, and MWCNT. Molecular docking analysis has been performed with (D-glucosamine) and targeted proteins (Protein Data Bank ID: 1HJV, 1ALU) to determine the binding affinity and interactions. Field emission scanning electron microscopy study showed that the composite KRT/CS/GLS incorporated on the surface of functionalized MWCNTs effectively. Fourier transform infrared spectroscopy analysis confirmed the presence of KRT/CS/GLS in the nanocomposite and remained intact. X-ray diffraction analysis indicated that the nature of the composite in MWCNT transformed from a crystalline to an amorphous state. Thermo gravimetric 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 (500 μg 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).