The Belgian playmaker reached the 100-goal landmark for the Citizens after scoring a brace in last weekend's 4-2 win over Crystal Palace in the Premier League.
However, Pep Guardiola has decided to omit De Bruyne - who is still working his way back to full fitness following a five-month injury layoff - from his starting lineup and has made a total of four changes to the side that beat the Eagles.
De Bruyne, Rico Lewis, Oscar Bobb and Julian Alvarez all move to the substitutes' bench, while Manuel Akanji, Phil Foden, Mateo Kovacic and Bernardo Silva all return to the starting lineup.
Stefan Ortega retains his place in goal even though Ederson has recovered from a muscle injury, while Josko Gvardiol is fit to start in the back four alongside Akanji, John Stones and Ruben Dias after shaking off a "niggle" that forced him to withdraw at half time against Palace.
Rodri and Kovacic will link arms in centre-midfield, while Bernardo, Foden and Jack Grealish will provide support in attack for Erling Haaland.
As for Real Madrid, Carlo Ancelotti has made two changes to the lineup that began the 2-0 La Liga win over Athletic Bilbao on March 31, with Vinicius Junior and Eduardo Camavinga coming in at the expense of Nacho Fernandez and ex-Man City man Brahim Diaz.
With Nacho dropping to the bench, Aurelien Tchouameni is set to start at centre-back alongside Antonio Rudiger, while Dani Carvajal and Ferland Mendy continue as full-backs.
Camavinga will join Federico Valverde and Toni Kroos in centre-midfield, while England star Jude Bellingham, who has scored four goals in six Champions League games this season, will link up with Vinicius and Rodrygo in attack.
Eder Militao has recovered from an ACL injury to make the matchday squad and will begin as a substitute, joining the likes of Luka Modric, Dani Ceballos and Arda Guler on the bench.
Real Madrid: Lunin; Carvajal, Tchouameni, Rudiger, Mendy; Valverde, Kroos, Camavinga; Bellingham; Rodrygo, Vinicius Jr
Subs: Kepa, Fran, Militao, Nacho, Modric, Joselu, Vazquez, Ceballos, Fran Garcia, Brahim, Guler
Manchester City: Ortega; Akanji, Stones, Dias, Gvardiol; Rodri, Kovacic; Bernardo, Foden, Grealish; Haaland
Subs: Ederson, Carson, Doku, De Bruyne, Alvarez, Gomez, Nunes, Bobb, Susoho, Lewis
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