The plans represent an almost doubling of the spending by Khan's predecessor Boris Johnson, but critics have questioned how the mayor will fund the investment, and why it will not help to expand the cycle hire scheme.
The investment will be laid out in the Transport for London business plan on 15 December.
In addition to future projects, cash will go towards a new cyclist and pedestrian bridge linking Rotherhithe and Canary Wharf, and completion of a cycle “superhighway” between Farringdon and Kings Cross as well as three new routes.
The mayor will also appoint a new walking and cycling commissioner for London to advocate active travel in the capital.
Khan said: “Making cycling safe and easier can provide huge benefits for us all – improving our health, cleaning up our toxic air, and helping tackle congestion.”
He has also committed to banning the most unsafe HGV lorries from London's streets by 2020 in order to make cycling safer in the capital.
However, London Conservatives transport spokesman Keith Prince questioned how City Hall will afford the level of investment planned by Khan.
"Having already taken £640m from TfL's budget with his fares freeze, the mayor is now promising large-scale cycling investment without explaining where the money is coming from,” Prince said.
"We all want to see investment in cycling infrastructure in London. I just hope this is not yet another grand pledge on which the Mayor fails to deliver."
Similarly, Lib Dem assembly member Caroline Pidgeon argued that cycling plans had been underfunded by Johnson, and questioned why the mayor's plans did not include expansion of the city's bike rental scheme.
“Extending the cycle hire scheme into places such as Richmond and Rotherhithe should be key priorities of any Mayor who is truly serious about cutting congestion and making London a cycling city for everyone,” Pidgeon said.