A self-designed Ti-35421 (Ti-3Al-5Mo-4Cr-2Zr-1Fe wt%) titanium alloy is a new type of low-cost high strength titanium alloy. In order to understand the hot deformation behavior of Ti-35421 alloy, isothermal compression tests were carried out under a deformation temperature range of 750–930 °C with a strain rate range of 0.01–10 s−1 in this study. Electron backscatter diffraction (EBSD) was used to characterize the microstructure prior to and post hot deformation. The results show that the stress–strain curves have obvious yielding behavior at a high strain rate (>0.1 s−1). As the deformation temperature increases and the strain rate decreases, the α phase content gradually decreases in the α + β phase region. Meanwhile, spheroidization and precipitation of α phase are prone to occur in the α + β phase region. From the EBSD analysis, the volume fraction of recrystallized grains was very low, so dynamic recovery (DRV) is the dominant deformation mechanism of Ti-35421 alloy. In addition to DRV, Ti-35421 alloy is more likely to occur in continuous dynamic recrystallization (CDRX) than discontinuous dynamic recrystallization (DDRX).