This study utilizes C60 fullerene to prepare a new carrier (Buckysomes) for pemetrexed (for the first time) and investigate the contribution of such conjugates on the behavior, release, and anticancer activity against lung cancer cells through cell line study. Methods: fullerene C60 was synthesized by laser irradiation of polycyclic hydrocarbons (PAHs) and loaded with pemetrexed (PMX). The pemetrexed loaded on fullerene C60 was evaluated by fourier transform infrared spectroscopy (FTIR), power x-Ray diffraction (PXRD), scanning electron microscopy (SEM), atomic force microscopy (AFM), differential scanning calorimetry (DSC), particle size analyzer, and zeta potential. The percentage of drug loading, in vitro release, and anticancer activity against lung cells A549 were also evaluated. Results: Characterization technique showed the successful loading of pemetrexed on fullerene C60 nanocarrier, S9 was the optimum sample with percentage of yield reach to 90.17. Release profile shows that the percentage release of pemetrexed after 240 minutes equal 40% from pure PMX, while the release after 240 minutes was found to be 55% from pemetrexed loaded fullerene C60 nanocarrier, it is reached to 91.7% for pemetrexed loaded on fullerene nanocarrier versus 72% for pemetrexed at the end of 24 hours release. pemetrexed loaded on fullerene nanocarrier (S9) shows lower IC50 (1.03 μM) and higher cytotoxic effect than that of pure PMX (3.1 μM) and blank fullerene (75.5 μM) on A549 cancer cell at different times of exposure (24, 48, and 72 hours). The anticancer activity of pemetrexed against lung cancer cells A549 was improved upon loading on fullerene nanocarrier (80% cell death for S9) in comparison with the free drug (60% cell death) at 24 hours, although the fullerene showed limited anticancer activity (20% cell death after 24 hours) indicating that the loading process led to improve the cytotoxic activity of the drug for the tumor cells. Pemetrexed loaded on fullerene nanocarrier (S9) at 24 hours shows a higher percentage of cell death (52%) than the native pemetrexed (35%), this could be attributed to the synergistic effect of fullerene on the cytotoxic effect of pemetrexed in its complex (S9) which may lead to faster onset of action Conclusion: This work succeeded to prepare Buckysomes for pemetrexed using fullerene C60 that may act as a promising nanocarrier for pemetrexed since it produced enhancement of anticancer activity, solubility and release profile.