The second siege of Takatenjin came only six years after Takeda Katsuyori took the fortress. This second siege lasted from 1580 until 22 March 1581, the siege ended within four months and ended with the deaths of 680 of Okabe Motonobu's garrison.
In 1580, during the conflict between Tokugawa clan against Takeda clan, Tokugawa Ieyasu built up around five fortresses in order to isolate the Takatenjin castle from external supplies and reinforcements. Those new fortresses consisted of:
In addition of those 5 new fortresses, Ieyasu renovated an old castle located 4 km north of Takatenjin castle. This castle were named Ogasayama fort, which originally built by Ieyasu far before during the conquest of Tà Âtà Âmi Province against Imagawa clan to capture Kakegawa Castle. As these six fortresses completed, Ieyasu assigned Ishikawa Yasumichi to garrison the Ogasayama fort, Honda Yasushige to garrison Nogasaka fort, Osuga Yasutaka to garrison Higamine fort, Shishigahana Fort & Nakamura Fort, while Sakai Ietada was appointed to garrison Mitsuiyama fort.
With the completion of those cluster of castles which referred as "six fortress of Takatenjin", the Takatenjin castle which defended by Okabe Motonobu immediately suffered starvations as they deprived any sources or supply to sustain the population inside the castle, while the siege outside the castle by Oda-Tokugawa forces intensified. In response, Motonobu tried to negotiate truce with Ieyasu by offering Takisakai Castle and Koyama Castle to be given for Ieyasu in exchange that Takatenjin castle spared from the siege. However, Oda Nobunaga refused the plea from Motonobu.
On January 3, 1581, Ieyasu was informed that a force under the command of Takeda Katsuyori was approaching. Oda Nobunaga got the same information and he immediately sent Oda reinforcements led by Mizuno Tadashige.
In March 1581, Takatenjin Castle ran out of food, and the castle's soldiers ate plants to survive hunger. Motonobu decided that he should break the siege and giving a party with his remaining soldiers at night before they goes out to try launch assault against Ieyasu's blockade. Shortly after 10 pm. on March 22nd, Motonobu led his remaining soldiers in an assault on Mitsuiyama fort which defended by Ishikawa Yasumichi, as it was considered to be the weakest fortress among the six Tokugawa blockade fortress. However, as they launched the attack towards Mitsuiyama, Yasumichi immediately reinforced by à Âkubo Tadayo and Osuga Yasutaka, and resulted in a fierce battle, where Motonobu killed. Meanwhile, as the battle has phased with the army of Motonobu routed, à Âkubo Tadataka led a detachment to pursue any of fleeing enemies.
The siege was quite advantageous for Oda Nobunaga, as it significantly weakened his enemy, the Takeda clan; the Battle of Tenmokuzan the following year would come to be known as Takeda Katsuyori's last stand and the fall of the distinguished Takeda of Kai.
According to the Shinchà  Kà Âki by à Âta Gyà «ichi, "in all, 688 soldiers of various rank joined the commander of the castle in a desperate attack out onto the besiegers, and they were killed".
In Matsudaira Ietada (Fukà Âzu)âÂÂs diary Ietada nikki, he wrote that; âÂÂamong our own we lost about 130 in total, while the enemy lost more than 600âÂÂ.