Police fired teargas and violently removed the large group gathered in the capital. "I chose to die for Zimbabwe", sang some protesters, in the Shona language, before running away from clouds of tear gas.
"The police have cordoned off all of Harare but in court they were saying they don't have enough manpower to supervise our demonstration". One woman was beaten by police and is lying on the street.
The party's spokesman, Daniel Molokele, said, however, it would go ahead with protests outside Harare planned for next week.
An attempt by the Movement for Democratic Change (MDC) party to challenge the ban in court was rejected, but more than 100 MDC supporters defied the ruling before being chased by baton-wielding officers from a city square.
In January, a violent security crackdown in Harare against demonstrations over fuel prices and supply left more than a dozen people dead.
The opposition party called off the march after failing to overturn a police ban of the demonstrations at the High Court in the morning, but hundreds of its supporters were already in the CBD.
"Do not take part, you will rot in jail", shouted police officers through megaphones on Thursday in downtown Harare and some residential areas.
Africa Unity Square, overlooking the Zimbabwean parliament, is where crowds gathered in November 2017 to demand Mugabe, the country's long-time autocratic ruler, step down.
Police had warned to use punitive actions against protesters, under various provisions of public order and security.
To avoid any possible trouble, many residents avoided going out.
The MDC, which disputes President Emmerson Mnangagwa's presidential election win a year ago, called the protest against the government's handling of the economy, which is mired in its worst crisis in a decade.
The demonstration, dubbed "Free Zimbabwe March" is scheduled for this Friday in Harare before spilling to other major towns and cities.
Anger is mounting as Zimbabweans grapple with soaring inflation, rolling power cuts, and shortages of USA dollars, fuel and bread.
On Thursday, the president urged the opposition to engage in dialogue.
Armed police had erected barricades across around the city early Friday in a bid to deter protesters and searched cars, buses and pedestrians for weapons.
The series of demonstrations has been widely viewed as a test of how Mnangagwa, who this year so far has failed to make good on promises of political and economic reform, responds to dissent in a country tainted by a long history of repression. Undeterred, the demonstrators overran the police and went marching through CBD singing and chanting slogans denouncing President Emmerson Mnangagwa's government, which they accused of fomenting economic instability and hunger that stalks numerous jobless Zimbabweans.
In addition to the debilitating shortages of power and water, about a third of Zimbabwe's 15 million people are in dire need of food aid, according to aid agencies.
Amnesty International on Friday accused Zimbabwe's president of repeating some of the brutal tactics as employed by the former government of Robert Mugabe.