The 58-year-old from Bamber Bridge will cycle 250 miles in three days in tribute to Jill Brakewell, who died of leukaemia last November.
He will start his journey in London later this month, finishing in Ghent, in Belgium.
All money raised will go towards for Rosemere Cancer Foundation, which supported 64-year-old Jill before she died.
John, a regional development manager for housing group Places for People, said: “I am not a natural cyclist but last year, I cycled 1,000 miles in 10 days to Cannes to raise money for North West Air Ambulance.
“This year, my quest is a personal one as it is to raise money for a cause close to my sister’s heart.”
The father-of-three will be accompanied en route by Jane Kennett, a former Royal Preston Hospital nurse, who now works as a medical researcher in London. She was the organiser of last year’s Cannes expedition and has plotted the route to Ghent.
A number of cyclists will join them to pedal some of the way and act as support.
John is hoping his cycle will raise around £1,500 for Rosemere, with his fund-raising having been kick-started by a pledge of support from new Bamber Bridge gym, Places Gym.
The grandfather-of-three has already helped raise £6,000 in his sister’s name for the charity after he, Jill’s widower Mick Brakewell and their children Helen Duchamp, David and Roger Brakewell, organised a music and laughter night at Bamber Bridge Catholic Club in March.
To sponsor John, visit http://www.justgiving.com/John-Wright46.