The hosts narrowly overcame Reading 4-3 on penalties in the first round of the competition, while this will be the visitors' first EFL Cup match of the season.
Match preview
Colchester's stay in the fourth division of English football was extended for a ninth season as they narrowly placed in 22nd in 2023-24, just three points above the drop zone with 45 points.
This represents another decline for The U's, who have fallen from competing for a playoff spot in 2016-17, 2018-19, and 2019-20 to battling against relegation in 2020-21, 2022-23, and 2023-24.
Danny Cowley, who took charge of the side in January 2024, will be aiming to guide the team clear of the drop zone this campaign, and they have started the season in reasonable form.
Cowley's side have experienced a mixed start to the season with a disappointing 4-2 loss to AFC Wimbledon after leading 2-0 within nine minutes, followed by a notable 4-3 penalty win over Reading in the EFL Cup, a 2-0 victory against MK Dons, and a 1-0 defeat to Harrogate Town.
Despite their recent loss to Harrogate, Colchester's strong performance, which saw them hold 63% possession and take 24 shots compared to their opponents' eight, indicates plenty of promising signs for Cowley's side.
They will now face a challenging task against Premier League side Brentford, who have kicked off their top division campaign with one win and one loss from two games.
Thomas Frank's side have established themselves as a consistent Premier League team since their promotion as Championship playoff winners in 2020-21, placing 13th, ninth and 16th.
Last year was undoubtedly Brentford's most difficult year in the top flight, finishing with 20 points fewer than the previous year with just 39 points, though they still comfortably cleared the relegation zone by 13 points.
Frank's men also endured a tough pre-season in preparation for the new term, triumphing in just one of their five friendly fixtures, but they have had a positive start to the 2024-25 season.
The Bees defeated Crystal Palace 2-1 in a strong opening performance to the new Premier League campaign, followed by a 2-0 defeat to Liverpool in their most recent outing.
Ahead of a challenging run of fixtures that will match Brentford against Southampton, Manchester City, Tottenham Hotspur and West Ham United, Frank will be hoping his side can secure a confidence-boosting victory against Colchester in this one.
Colchester United EFL Cup form:
W
Colchester United form (all competitions):
Brentford form (all competitions):
Team News
Ben Goodliffe scored both goals in Colchester's eventual 4-2 defeat to Wimbledon on the opening day of the season but was forced off in the 60th minute due to an injury, and the defender is unlikely to feature here.
Despite losing in their most recent match, a promising performance could leave Cowley opting to name a similar starting side against Brentford on Wednesday.
Brentford are anticipated to be without Aaron Hickey (hamstring), Thiago Rodrigues (meniscus), Rico Henry (knee) and Joshua Dasilva due to injury issues.
Elsewhere, Frank is likely to rotate his squad before his team host Southampton in the Premier League at the weekend, potentially giving Fabio Carvalho and Sepp van den Berg their first starts for the club.
Colchester United possible starting lineup:
Macey; Hunt, Flanagan, Iandolo; Egbo, Bishop, Woodyard, Payne, Gordon; Hopper, Tovide
Brentford possible starting lineup:
Flekken; Roerslev, Mee, Van den Berg, Ajer; Carvalho, Norgaard, Onyeka, Damsgaard; Schade, Wissa