The fifth and final Monument of the season – the 'Race of the Falling Leaves' – will be fought over 241km through the Alps-region between Bergamo and Como in northern Italy.
Movistar rider Valverde will be challenged by the same rivals who were beaten for the world championships road race crown in Innsbruck last month, including France's Romain Bardet who finished second and third-placed Canadian Michael Woods.
“Being world champion is important for whoever it might be,” said Valverde.
“It's also a race that I like a lot and which I've always nearly won.”
Valverde finished second in the Tour of Lombardy in 2013 and 2014.
The 38-year-old warmed up by finishing third in the Milan-Turin semi-classic on Wednesday, which was won by French rider Thibaut Pinot (Groupama-FDJ), who will be among the favourites this weekend.
Valverde, who was banned for two years following Spanish doping probe Operacion Puerto, this week defended his record since returning in 2012.
“That's water under the bridge, and I don't even want to talk about it. I believe I've sufficiently demonstrated who I am since then.”
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Italian Vincenzo Nibali will be defending his title, over a route that is virtually identical to last year's apart from a final three kilometre drop to the finish line at Como.
Nibali, of the Bahrain-Merida team, also won the race in 2015, and is one of two former winners competing along with 2014 champion Daniel Martin of Ireland.
The 33-year-old Nibali, who has won all the Grand Tours including the Tour de France in 2014, but has struggled for form this season.
Race favourites include French duo Pinto, and Romain Barden, Italian Gianni Moscon and Colombians Rigoberto Uran and Miguel Angleo Lopez.
“With the form I'm now showing I'm getting more and more convinced of continuing the positive streak until Saturday in Lombardy,” said Pinot after winning the Milan-Turin.
British Sky riders Chris Froome, winner of the Giro d'Italia, and Tour de France winner Geraint Thomas, will not be competing.
The Mitchelton-Scott team are led by 2018 Vuelta champion Simon Yates and his twin brother Adam.
The route features iconic climbs up Madonna del Ghisallo and Muro di Sormano, with the final hill, a 1.7km ascent to Monte Olimpino with a five percent gradient.