Nearly 200 companies and 80 individuals in third countries such as China, Belgium, and Slovakia, which allegedly support Russia in the procurement of materials for its weapons programme, are affected, it added on Wednesday.
The sanctions are also aimed at Russia's production of chemical and biological weapons.
"Today’s actions will further disrupt and degrade Russia’s war efforts by going after its military-industrial base and the evasion networks that help supply it," US Treasury Secretary Janet Yellen said.
On Wednesday, the US State Department accused Moscow of using various irritant gases during its all-out war against Ukraine, which violates the United Nations' Chemical Weapons Convention.
Three people who are reportedly linked to the death of Alexei Navalny are also being sanctioned. The Kremlin critic died in Russian custody in February.
Navalny, a long-standing and staunch opponent of Putin, died in a prison camp in the Arctic Circle in Siberia in February.
It has not been independently established whether the 47-year-old died naturally, as his death certificate says. However, his supporters say he was murdered.
As a result of the new sanctions, any assets of those affected in the US will be frozen.
US citizens or people who are in the US are prohibited from doing business with sanctioned companies and individuals.
The sanctions also make international business much more difficult for those affected.