{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,8,7]],"date-time":"2026-08-07T15:51:58Z","timestamp":1786117918884,"version":"build-2736575974"},"reference-count":56,"publisher":"American Society for Microbiology","issue":"8","license":[{"start":{"date-parts":[[2026,8,4]],"date-time":"2026-08-04T00:00:00Z","timestamp":1785801600000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"},{"start":{"date-parts":[[2026,8,4]],"date-time":"2026-08-04T00:00:00Z","timestamp":1785801600000},"content-version":"tdm","delay-in-days":0,"URL":"https:\/\/journals.asm.org\/non-commercial-tdm-license"}],"content-domain":{"domain":["journals.asm.org"],"crossmark-restriction":true},"short-container-title":["Microbiol Spectr"],"published-print":{"date-parts":[[2026,8,4]]},"abstract":"<jats:title>ABSTRACT<\/jats:title>\n                  <jats:sec>\n                    <jats:title\/>\n                    <jats:p>\n                      The ongoing spread of the monkeypox outbreak underscores the urgent need for the development of effective therapeutics against the monkeypox virus (MPXV). Aminopterin is a classical antifolate agent known for its anti-tumor activity through the inhibition of folate metabolism. Here, aminopterin was identified as an effective anti-MPXV agent. It significantly inhibited MPXV replication, unlike other antifolate agents such as methotrexate, pemetrexed, and LSN3213218. In Vero E6 cells, the half-maximal inhibitory concentration (IC\u2085\u2080) was 36.10 nM, the half-cytotoxic concentration (CC\u2085\u2080) was 3,310 nM, and the selectivity index (SI) was 91.690. In human foreskin fibroblast cells, the IC\n                      <jats:sub>50<\/jats:sub>\n                      was 32.73 nM, CC\u2085\u2080 exceeded 10 \u03bcM, and SI was greater than 305.530.\n                      <jats:italic toggle=\"yes\">In vivo<\/jats:italic>\n                      , aminopterin not only inhibited MPXV infection but also mitigated high-dose viral infection-associated inflammation. Furthermore, the data indicated that aminopterin reduced viral load by interfering with viral attachment and replication. Together, these findings identify aminopterin as a promising candidate for anti-MPXV drug development.\n                    <\/jats:p>\n                  <\/jats:sec>\n                  <jats:sec>\n                    <jats:title>IMPORTANCE<\/jats:title>\n                    <jats:p>\n                      The urgent need for effective monkeypox virus (MPXV) therapeutics is underscored by the ongoing outbreak. This study identifies aminopterin as a potent anti\u2011MPXV agent among four antifolate agents. Crucially, aminopterin suppresses viral infection and mitigates infection-associated inflammation\n                      <jats:italic toggle=\"yes\">in vivo<\/jats:italic>\n                      . These findings position aminopterin as a potential candidate for the development of an antiviral agent against MPXV.\n                    <\/jats:p>\n                  <\/jats:sec>","DOI":"10.1128\/spectrum.01581-25","type":"journal-article","created":{"date-parts":[[2026,6,22]],"date-time":"2026-06-22T12:00:42Z","timestamp":1782129642000},"update-policy":"https:\/\/doi.org\/10.1128\/asmj-crossmark-policy-page","source":"Crossref","is-referenced-by-count":0,"title":["Antifolate agent aminopterin demonstrates potent anti-monkeypox virus activity\n                    <i>in vitro<\/i>\n                    and\n                    <i>in vivo<\/i>"],"prefix":"10.1128","volume":"14","author":[{"ORCID":"https:\/\/orcid.org\/0000-0001-9111-3139","authenticated-orcid":false,"given":"Jing","family":"Wu","sequence":"first","affiliation":[{"name":"Interdisciplinary Eye Research Institute (EYE-X Institute), Bengbu Medical University","place":["Bengbu, China"]},{"name":"Key Laboratory of Infection 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