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Martensitic high-strength steels are prone to exhibit premature fatigue failure due to fatigue crack nucleation at non-metallic inclusions and other microstructural defects. This study investigates the fatigue crack nucleation behavior of the martensitic steel SAE 4150 at different microstructural defects by means of micromechanical simulations. Inclusion statistics based on experimental data serve as a reference for the identification of failure-relevant inclusions and defects for the material of interest. A comprehensive numerical design of experiment was performed to systematically assess the influencing parameters of the microstructural defects with respect to their fatigue crack nucleation potential. In particular, the effects of defect type, inclusion–matrix interface configuration, defect size, defect shape and defect alignment to loading axis on fatigue damage behavior were studied and discussed in detail. To account for the evolution of residual stresses around inclusions due to previous heat treatments of the material, an elasto-plastic extension of the micromechanical model is proposed. The non-local Fatemi–Socie parameter was used in this study to quantify the fatigue crack nucleation potential. The numerical results of the study exhibit a loading level-dependent damage potential of the different inclusion–matrix configurations and a fundamental influence of the alignment of specific defect types to the loading axis. These results illustrate that the micromechanical model can quantitatively evaluate the different defects, which can make a valuable contribution to the comparison of different material grades in the future.