Gers have made the 46-year-old former Ibrox midfielder their number one target to replace Pedro Caixinha.
But in club statement Aberdeen said: “Aberdeen Football Club has announced this evening that Rangers have contacted the Club asking for permission to speak to Derek McInnes and that permission has been refused.”
Ibrox chiefs have been accused of dithering over the six weeks that have followed Caixinha’s sacking.
But they finally made their move for McInnes late on Tuesday afternoon – just two days after he had seen his Dons side beaten for the second time in the space of a week by the Light Blues.
The protracted recruitment process and associated rumour mill has frustrated the former St Johnstone and Bristol City manager.
After Sunday’s 2-1 defeat, McInnes admitted his annoyance that speculation linking him to the Govan vacancy had been allowed to “drag on and on”.
It now remains to be seen whether Gers will return with an improved offer, with McInnes understood to have an £800,000 release clause written into his Pittodrie contract.
However, Dons chairman Stewart Milne will hope this is the end of the matter. Aberdeen travel to Dundee on Friday night but Milne will be not want his club’s preparation for that clash to again be overshadowed by events in Glasgow.
Meanwhile, the Ibrox squad are readying themselves to prove they deserve to play a part in the club’s future, according to midfielder Jason Holt.
The former Hearts battler forced himself back into the Gers line-up under caretaker boss Graeme Murty but knows he will have to justify his involvement again once a new manager is appointed.
“It’s great to be back in the team playing every week and helping the boys,” he said as the Rangers players made their annual Christmas visit to the Royal Hospital for Children in Glasgow. “It’s not just been myself, the whole squad have enjoyed working under Graeme.
“Graeme came in to help us all out and we’ve given him 100 per cent.
“But as for a new manager, everyone will be playing to prove that they deserve to be in the team.”
Rangers’ back-to-back wins over Aberdeen has pushed them up to second spot but they now return to the task of winning three games on the bounce – a feat Gers have not managed since last December.
“The last two results have been really good for the club but it’s about pushing on now and building on it,” added Holt. “It was important that we showed a reaction to the two defeats to Hamilton and Dundee as they were hugely disappointing.
“It was a great result but it will count for nothing if we don’t back it up.
“We won’t get carried away. It’s only two games we have won and now we need to build on that as we’ve shown this season and last that consistency has been a problem for us.”
The Scottish Football Association has confirmed it has received Gers’ appeal against Ryan Jack’s weekend dismissal. His case will be heard at a hearing on Thursday morning.