The top court set the April 1 deadline in December barring liquor shops from operating within a range of 500 meters (1,640 feet) from India's highways, where 17 traffic accident deaths occur every hour.
On Friday the court dismissed a slew of petitions seeking a reprieve on the ban, including from state governments which had sought more time to implement the order.
However, the court extended the ban to pubs, bars and restaurants near highways, with the government estimated to lose billions of rupees in taxes.
Last year the court told the government not to renew existing licenses after March 31 and banned issuance of fresh licences in the prohibited area.
It also ordered the removal of all liquor banners and advertisements from such routes.
India, which has a network of 250,000 kilometres (155,000 miles) of national and state highways, tops the world's deadliest roads. Nearly 150,000 people were killed in 2015, according to the ministry of road transport and highways.
Of those, 6,755 deaths were caused due to drunken driving, the ministry said.
The government has launched fresh initiatives to make roads safer, including revamping infrastructure and proposed stringent laws.
Under a new proposed law drunken drivers would be fined rupees 10,000 ($154) up from 500 ($7) and handed jail terms ranging from three to six months.
Harman Singh Sidhu of Arrive Safe NGO, one of the petitioner's in the liquor ban, told AFP that it was a historic ruling that had led to the closure of nearly 48,000 liquor shops.
"It is certainly going to save lots of lives and bring awareness to the public about this health hazard," Sidhu said.
India owns just one percent of the world's vehicles but accounts for 15 percent of global traffic deaths, according to the World Bank.
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