2020/09/16 | Reinmeer Aomori | | 2-1 | | Sapporo University | | | | |
| Tonan Maebashi | | 2-3 | | Vonds Ichihara | | | | |
| Iwaki | | 4-0 | | Oyama SC | | | | |
| Kanazawa Seiryo University | | 1-2 | | Toyama Shinjo | | | | |
| Ococias Kyoto | | 0-1 | | Nara | | | | |
| Tokyo Musashino | | 3-3 (5-4 Pen.) | | Tochigi City | | | | |
| National Institute of Fitness | | 1-0 | | Okinawa SV | | | | |
| Cento Cuore Harima | | 1-1 (2-4 Pen.) | | MIO Biwako Shiga | | | | |
| Verspah Oita | | 2-2 (5-4 Pen.) | | Tegevajaro Miyazaki | | | | |
| Toin University | | 4-2 | | Yamanashi Gakuin University | | | | |
| University of Tsukuba | | 2-0 | | Tokyo Int. University | | | | |
| Maruyasu Okazaki | | 3-2 | | Suzuka Point Getters | | | | |
| Yonago Genki | | 0-5 | | Kagura Shimane | | | | |
| EV Internacional | | 0-4 | | MD Nagasaki | | | | |
| Kumamoto Teachers Club | | 0-0 (1-3 Pen.) | | Fukuoka University | | | | |
| Sony Sendai | | 3-0 | | Saruta Kogyo | | | | |