Al Ain, who won the maiden title in 2003 after beating Thailand's BEC Tero Sasana, opened their campaign at the end of February with an impressive 3-1 home win over two-time continental champions Al Hilal thanks to goals from Omar Abdulrahman, Alex Brosque and UAE Pro League top scorer Asamoah Gyan.
And after Cosmin Olaroiu's side extended their lead at the top of the UAE Pro League table to eight points with a 2-1 win over Al Wasl on Thursday thanks to a double from Ghanaian striker Gyan, Al Ain will be full of confidence ahead of their trip to Tehran.
"Our philosophy is to face each match separately. We achieved a hard-fought victory against Al Wasl despite the fact that the players were tired and they were affected with fatigue," said Al Ain's Romanian coach Olaroiu.
"We are looking forward to the match against Esteghlal and our last win in the local league will give us a big push.
"We have a heavy schedule as we will be playing three matches in seven days and hopefully the players will not be affected."
Esteghlal, though, will also be looking to build on opening their AFC Champions League campaign with a thrilling 3-3 draw against Al Rayyan in Qatar after Amir Ghalenoei's side stretched their lead at top of the Iran Pro League table to three points with a 1-0 win over Tractorsazi Tabriz on Friday.
Iran international Javad Nekonam, who also found the net against Al Rayyan two week ago, came off the bench to net the first half winner as Esteghlal edged out their fellow AFC Champions League qualifiers at Azadi Stadium despite playing over an hour with 10 men.
Also in Group D on Wednesday, two-time Asian Club Championship winners Al Hilal will entertain Al Rayyan without Salem Al Dawsari after the midfielder was sent-off against Al Ain.
In Group C, new Al Ahli coach Aleksandar Ilic will take his new charges to Dubai to face Al Nasr after beginning his reign with a 5-1 win over Narjan in the Saudi Pro League on Friday.
Despite 2012 finalists Al Ahli beginning their new continental campaign with a 2-0 home win over Qatar's Al Gharafa, Czech coach Karel Jarolim parted company with the King's Cup champions to be replaced by Serbian Ilic.
Play-off winners Al Nasr began their camping with a 3-0 defeat by Iran champions Sepahan, who travel to Qatar to face Al Gharafa on Wednesday.
In the East, China's Beijing Guoan entertain J.League champions Sanfrecce Hiroshima in Group G, while former champions Pohang Steelers travel to Tashkent to face two-time semi-finalists Bunyodkor.
With Beijing looking to advance past the group stage for just the second time, Aleksandar Stanojevic's side made an encouraging start two weeks ago with a goalless draw in Korea against Pohang before beginning the new Chinese Super League season with a timely 4-1 win over Shanghai East Asia.
Sanfrecce travelled to Beijing after registering a first win of the year at the weekend after beating Albirex Niigata 2-1 in the J.League having suffered a 2-0 home defeat by Bunyodkor two weeks ago.
Pohang also picked up their first three points of 2013 at the weekend with a 3-0 win over Daejeon Citizen's in the K-League, while Bunyodkor's strong start to the season continued with a 3-0 win over Nasaf in the Uzbek League on Friday.
In Group H, Korea Republic's Suwon Samsung Bluewings entertain China's Guizhou Renhe, while Australia's Central Coast Mariners make the trip to Japan to face Kashiwa Reysol.
Two weeks ago, Suwon shared a goalless draw with Central Coast and Brazilian striker Cleo scored on the stroke of half-time as Emperor's Cup winners Kashiwa edged out Guizhou in China.
On Saturday, 2011 semi-finalists Suwon maintained a perfect start to the new K-League Classic season with a 1-0 win over Gangwon, while AFC Champions League debutants Guizhou Renhe suffered a 2-1 defeat by Qingdao in their Chinese Super League opener.
Kashiwa will look to bounce back from a 3-0 defeat by FC Tokyo at the weekend, while A-League Premiers Central Coast are out to end a three game winless streak having followed their draw with Suwon with defeats by Western Sydney Wanderers and Sydney in the A-League.
Wednesday's AFC Champions League Fixtures
Al Gharafa vs Sepahan
Al Nasr vs Al Ahli
Esteghlal vs Al Ain
Al Hilal vs Al Rayyan
Beijing Guoan vs Sanfrecce Hiroshima
Bunyodkor vs Pohang Steelers
Kashiwa Reysol vs Central Coast Mariners
Suwon Samsung Bluewings vs Guizhou Renhe