The announcement was made today in Abu Dhabi by International Co-operation Minister Julian Fantino at the Global Vaccine Summit.
The amount represents a 41 per cent increase compared to the average amount contributed per year to polio eradication since 2006.
The $250 million will primarily support efforts to eradicate polio in three countries where it remains a persistent problem: Afghanistan, Nigeria, and Pakistan.
The Global Polio Eradication Initiative was launched in 1988 when polio paralyzed more than 1,000 children each day in more than 125 countries.
Since then, at least 2.5 billion children have been immunized and the number of cases has dropped from about 350,000 in 1988 to 233 as of last year.
"Immunization saves the lives of millions of children every year, and is one of the most cost-effective health investments," said Fantino in a statement.
He also said there are challenges to getting vaccinations to those who need them and called on religious, government and community leaders to denounce attacks on immunization workers and to "continue promoting science-based information about this important work."
Among those in attendance at the summit was Microsoft founder Bill Gates as co-chairman of the Bill & Melinda Gates Foundation.
"Canada has been a long-time leader in achieving a polio-free world and making sure children get the vaccines they need no matter where they live,” said Gates.
"Canada's increased support over the next six years will help ensure we can end polio and build the infrastructure needed to help all children live healthy and productive lives."