Staff from train operator London Midland will be swapping rail tracks for roads this weekend.
A team of 61 is tackling the Vélo Birmingham cycling sportive to raise money for Birmingham Children's Hospital Charity.
The 100-mile, closed road ride starts in the city centre on Sunday, September 24, before taking in routes through the Black Country, Worcestershire and Staffordshire and returning to the finish line in Broad Street.
London Midland has worked closely with the children's hospital over the last year, acting as a presenting partner of the Big Sleuth 2017, the public art sculpture trail which has seen colourful bears taking up residence across the West Midlands.
Leading the London Midland team will be managing director Patrick Verwer and finance and contracts director Ian McLaren.
Mr Verwer said: "London Midland believes passionately in getting closer to the communities we serve.
"One of the ways we do this is by promoting and encouraging healthy lifestyles.
"With a 61-strong team of London Midland riders in this year's event, Vélo is a perfect partner for us.
"We hope everyone, whether taking part or spectating, has a fabulous day getting to explore their local area and cheering on the riders."
London Midland has advised that, while all stations will be open as usual on Sunday, rail passengers should allow extra time when travelling to and from stations on the Vélo route to account for road alterations.
Stourbridge Junction station will only be accessible to pedestrians from 6.30am to 11am.
The sportive is the flagship event of the Birmingham Cycle Revolution, a city council initiative aiming to make cycling an everyday way to travel in Birmingham over the next 20 years.