We evaluated the impact of the centralisation of three acute stroke units into a single hyperacute stroke unit within a large urban and rural NHS trust in North East England in June 2015. We performed retrospective interrupted time series analyses of 4,305 stroke patients admitted between 1 April 2013 and 31 December 2017 utilising data recorded for the Sentinel Stroke National Audit Programme. Centralisation was associated with reductions in total length of inpatient stay (–4.9 days [95% CI: –8.1 to –1.7]). Time from admission to thrombolysis shortened by 26.0 minutes (95% CI: –40.0 to –12.1), and time from admission to brain imaging for thrombolysed patients improved by 16.2 minutes (95% CI: –22.0 to –10.4). Time from stroke onset to hospital admission, mortality and dependency (as measured by median modified Rankin Scale) were unaffected by centralisation. This study provides further evidence to support the centralisation of acute stroke services in England.