The Interoceanic Highway stretches from the Pacific Ocean to the Atlantic, increasing accessibility to many areas of the Amazon. While it offers opportunities for economic growth and wellbeing, the highway has also prompted concerns about deforestation since its completion in 2010. Madre de Dios, in southeastern Peru, is considered a biodiversity hotspot and is facing issues related to intensifying agriculture and illegal gold mining. This project utilized Landsat 5 ETM, Landsat 8 OLI/TIRS, and Aqua and Terra MODIS data to analyze changes in land cover occurring surrounding the Interoceanic Highway in Tambopata, a province within Madre de Dios. To assess these changes, we analyzed the Normalized Difference Vegetation Index (NDVI) between 2001 and 2018, and conducted supervised classifications of the study area. The project found that forest cover in the region overall has decreased from 92.9% in 2001 to 85.0% in 2018, with larger changes occurring closer to the highway. In a buffer zone extending 250m from the highway, forest cover decreased from 65.4% in 2001 to 32.5% in 2018. Land cover change analyses demonstrated that most forest loss occurred through the conversion of forest to grassland.