BACKGROUND: Anti-TNF treatment failure in Crohn's disease is common, and the literature on the selection of subsequent treatment is scant. Ustekinumab may be associated with high persistence rates and it appears to be effective in perianal disease. AIMS: Primary objective: persistence, clinical, and biologic remission with ustekinumab. SECONDARY OBJECTIVES: Persistence of the first biologic therapy, reasons for change of treatment, need for dose optimization, surgery, hospitalizations, and adverse events with ustekinumab. METHODS: Retrospective, observational, single-center study from a prospective database of Crohn's disease adult patients receiving ustekinumab after failure of anti-TNF or vedolizumab. A sub-analysis was performed to evaluate ustekinumab persistence after the approval of risankizumab and upadacitinib. RESULTS: Mean duration with ustekinumab was 27.65 months (SD 18.27) and persistence was 86.76%. Clinical remission was 40.63% at week 4, 54.35% at week 8, 54.9% at year 1, 76.92% at year 4, and 100% at year 5. Persistence with ustekinumab was longer than with anti-TNF: year 1, 93.2 vs 72.06%; year 2, 89.4 vs 45.59%; and year 3, 86.1 vs 30.88%. Just over one-third (36.76%) of patients required dose optimization. Nine (13.24%) patients stopped treatment due to primary non-response [1 (1.47%)], loss of response [5(7.35%)], and adverse events [3 (4.41%)]. Eleven (16.18%) patients needed surgery and hospitalization. After the approval of upadacitinib and risankizumab, ustekinumab persistence was 80.88%. Seven (70%) of the patients with perianal disease achieved clinical remission and 4 (40%) completed fistula healing. CONCLUSIONS: Ustekinumab may have better persistence as a second-line treatment compared to anti-TNF and may be effective in perianal disease.