Dave Sowerby, who owns Sowerby Bros Cycle Shop, Princess Street, Mirfield, lost his wife Lynda, 41, to cancer just before Christmas in 2006.
Last year he and 24 other cyclists raised nearly £9,000 for the hospice where she was cared for, with a 170 mile ride in her memory through North Yorkshire. Now Dave is hoping to raise about £5,000 when he completes the ride again on August 2.
He said: "We were really pleased with how it went last year and we feel we have to do it again - it's becoming an annual thing.
"It is a difficult ride – we are hoping to do it in about 12 hours – and it takes in the highest roads in each Yorkshire county. You can't do it without support."
Dave said the ride would start at 7.30am in Buckden in North Yorkshire, and would end at the top of Holme Moss.
He said: "Lynda would probably think I am stupid doing so many miles but she would be very proud. It's not just about the money – it's in her memory and people want to do it because of that too."
Dave and Lynda's son Luke, seven, is hoping to join the team for the last leg of the cycle at Holme Moss.
The team is looking for sponsorship and also volunteer drivers to transport the cyclists up to Buckden on the day.
To sponsor the team or volunteer to help, contact Dave on 01924 496873 or 07875 897845.