Voclosporin (Voc) shows a protective effect and strengthening of the intestinal barrier in experimental colitis. The study compared the efficacy of Voc with cyclosporine A (CsA) in an experimental model of colitis and in human PBMCs. Voc or CsA treatment improved colitis-associated weight loss and reduced intestinal inflammation as shown by miniendoscopy, histopathology, and multiphoton endomicroscopy. Voclosporin significantly increased the barrier protein claudin 3 in the colon of mice with induced colitis. Voclosporin and CsA treatment of stimulated human PBMCs inhibited the activation of IL2-induced tyrosine kinase ITK, which triggers inflammation in IBD. Analyzes included immunofluorescence staining, flow cytometry, and cytokine profiling in murine lamina propria mononuclear cells. The results support Voclosporin as a potential therapy for acute intestinal inflammation. Voclosporin is a recently approved drug for lupus nephritis.