Second seed Elina Svitolina and Coco Gauff were beaten in the second round of the Eastbourne International.
Ukraine's Svitolina, 26, lost 6-4 7-6 (7-3) to French Open quarter-finalist Elena Rybakina of Kazakhstan.
American Gauff, 17, started strongly against lucky loser Anastasija Sevastova, but was defeated 4-6 7-5 6-2 by the 31-year-old Latvian.
It was a setback for Gauff the week before Wimbledon, where she had a fairytale run to the last 16 in 2019.
There was also disappointment for Canadian third seed Bianca Andreescu, who was beaten 6-3 6-3 by Estonia's Anett Kontaveit.
Another leading player to make an early exit was Swiss sixth seed Belinda Bencic, who was beaten 6-4 6-4 by compatriot Viktorija Golubic.
Belarusian top seed and 2018 finalist Aryna Sabalenka did advance, though, beating American Alison Riske 6-1 6-4.
There was British success in the men's doubles for Luke Bambridge and Jamie Murray, who beat French fourth seeds Fabrice Martin and Eduoard Roger-Vasselin 6-4 6-4.
Lloyd Glasspool, partnering Finland's Harri Heliovaara, defeated Australians Max Purcell and Luke Saville 6-2 6-3.
The Eastbourne International is the last grass-court stop in the UK before Wimbledon starts on Monday, 28 June.