Background: Individuals with cocaine use disorder experience heightened motivation to pursue rewards tied to cocaine, often triggered by associated cues. Cue reactivity and subsequent craving significantly elevate the risk of substance use, creating a pressing need for treatments that can help alleviate cravings. However, no pharmaceutical therapies for treating cocaine use disorder have been approved. Preclinical findings reveal dysfunctions in the glutamatergic pathway connecting prefrontal regions with the nucleus accumbens, which are correlated with cue-induced substance-seeking behaviour. These alterations, at both molecular and behavioural levels, can be reversed in rodents with Methods: Here, we assessed the impact of a short-term challenge with Results: In the placebo condition, participants showed increased cue reactivity towards cocaine pictures, accompanied by significantly higher cravings as compared to neutral images. In accordance with recent meta-analyses, cue reactivity was evident in parietal regions such as the posterior cingulate and precuneus, temporal regions like the hippocampus, the bilateral insula, and medial prefrontal regions, namely the inferior, middle, and superior frontal gyrus. Cue-induced activity in the superior frontal gyrus was strongly predicted by the individual duration of cocaine use. While Conclusions: Our findings show that prefrontal reactivity to cocaine cues can be reduced even by a brief pharmacological challenge with Clinical Trial Registration: https://clinicaltrials.gov, identifier NCT02626494.