Extinction Rebellion activists have blocked part of the Blackwall Tunnel in protest over the proposed Silvertown Tunnel.
Dozens of demonstrators blocked traffic on one of the tunnel's slip roads during Friday morning rush hour, unfurling a banner in the road.
Transport for London said the tunnel is still open, but warned drivers to "approach with caution and expect delays" after they blocked the northbound A102 exit slip road and the westbound A206.
The planned tunnel, set to open in six years time, has had fierce opposition locally.
Extinction Rebellion claim the tunnel will increase pollution and congestion and say the row has "become increasingly fierce" since a meeting with Deputy Mayor Heidi Alexander and Transport for London on August 2.
Greenwich Council is one of few councils to continuously back the new road, with Lewisham, as well as Hackney over the river, opposing it.
The tunnel will be paid for by a toll on the Blackwall Tunnel.
In a statement, the environmental group said: "We are demanding the cancellation of the Silvertown Tunnel. It is irresponsible and unaffordable in a climate emergency.
"We call on City Hall to do what they know is right. Sadiq Khan himself says he’s a champion of clean air and keen to address climate emergency – this is his big chance to put words into action for the future of Londoners and the planet. The decision is his to make.”
Victoria Rance, Silvertown Tunnel Campaign Coordinator, said: “We’re so grateful to Extinction Rebellion for taking this action today.
"We had a very frustrating meeting at City Hall on Friday with deputy mayor Heidi Alexander, who wasn’t prepared to listen.
"She had no plan for the climate emergency that she could tell us and their carbon budget is totally out of kilter with what needs to happen.
"We were left feeling there was nothing else left to do except take direct action like we have today.
"But we are writing to Sadiq Khan one more time with a detailed briefing from top climate experts in the hope he’ll listen. He has to listen.”
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A TfL spokesperson said: "The need for more river crossings in East London, to unlock growth and give residents and businesses better access to jobs and services, has been clear for decades.
"The Silvertown Tunnel, which is vital to support London's economy, has been designed to resolve the worsening congestion and pollution problem around Blackwall. It will improve overall air quality and enable new cross-river bus routes to be introduced.
“The traffic modelling demonstrates that overall traffic does not increase as a result of the Silvertown Tunnel scheme. This is because the user charge can be set to manage demand for the Silvertown and Blackwall tunnels. Currently around 100,000 vehicles use the Blackwall Tunnel without paying any charge.
"From October 2021, they will be required to meet the Ultra Low Emission Zone requirement or pay a daily charge. Once the Silvertown Tunnel opens they will also be required to pay a user charge for using the tunnel.
"We are absolutely committed to ensuring that the project is delivered with minimal impact to local residents. We will closely monitor noise and air quality during construction and traffic levels and emissions once the tunnel is complete to ensure the scheme delivers on these promises.”
