Ayo Olukoga’s deflected effort put the Rooks ahead on 19 minutes but Daniel Ajakaiye equalised just nine minutes later on his first Hornets start of 2024.
Chris Whelpdale restored Lewes’ lead on 38 minutes before Archie Tamplin secured all three points – and Sussex bragging rights – with two minutes to go.
The result leaves Horsham two points adrift of the play-offs in sixth, albeit with games in hand, while the Rooks are now four points and five places off the top five.
The Hornets meet SCFL Premier outfit Steyning Town in a Sussex Senior Cup semi-final at Lancing FC’s Culver Road on Tuesday evening.
Lewes, meanwhile, are next in action on Saturday when they entertain 14th-placed Canvey Island.
See some of John Lines pictures from the game on this page and the ones linked, and read Horsham manager Dominic Di Paola’s thoughts in the West Sussex County Times, out every Thursday.
1. Lewes FC end Horsham FC's unbeaten run to claim derby bragging rights - the match in pictures
Action from Horsham v Lewes Photo: John Lines
2. Lewes FC end Horsham FC's unbeaten run to claim derby bragging rights - the match in pictures
Action from Horsham v Lewes Photo: John Lines
3. Lewes FC end Horsham FC's unbeaten run to claim derby bragging rights - the match in pictures
Action from Horsham v Lewes Photo: John Lines
4. Lewes FC end Horsham FC's unbeaten run to claim derby bragging rights - the match in pictures
Action from Horsham v Lewes Photo: John Lines