{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2025,10,10]],"date-time":"2025-10-10T01:00:49Z","timestamp":1760058049901,"version":"build-2065373602"},"reference-count":31,"publisher":"MDPI AG","issue":"6","license":[{"start":{"date-parts":[[2025,3,17]],"date-time":"2025-03-17T00:00:00Z","timestamp":1742169600000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"DOI":"10.13039\/501100002241","name":"Japan Science and Technology Agency (JST)\/Japan International Cooperation Agency (JICA)\u2019s Science and Technology Research Partnership for Sustainable Development (SATREPS) Projects: \u201cValorization of Bioresources based on Scientific Evidence in Semi and Arid Land for Creation of New Industry\u201d","doi-asserted-by":"publisher","award":["JPMJSA1506"],"award-info":[{"award-number":["JPMJSA1506"]}],"id":[{"id":"10.13039\/501100002241","id-type":"DOI","asserted-by":"publisher"}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Molecules"],"abstract":"<jats:p>Tryptase, which is abundant in human mast cells and is involved in allergic inflammations such as asthma, is a serine protease. We isolated a tryptase inhibitor, procyanidin B8 3,3\u2032-di-O-gallate (PB8GG\u2019), a tannin, from the methanol extract of rhubarb (RHEI RHIDOMA), which is a traditional Chinese medicine (Kampo medicine in Japan). Since it did not inhibit another serine protease trypsin, PB8GG\u2019 specifically inhibited tryptase. A standard kinetic analysis of the inhibitory fashion of PB8GG\u2019 against tryptase suggested that PB8GG\u2019 inhibited tryptase in an allosteric manner. We searched for other tannins like PB8GG\u2019 expected to bind tryptase using AutoDock vina. Two ellagitannins, carpinins B and E, isolated from young leaves of Carpinus japonica were selected as candidates of tryptase inhibitors. Carpinins B and E themselves had almost no inhibitory activity against tryptase but enhanced the inhibitory activity of PB8GG\u2019 against tryptase. This is an example that shows that a combination of an allosteric inhibitor with other compounds that bind but have no inhibitory activity can be used to develop a clinically useful combinatorial enzyme inhibitor.<\/jats:p>","DOI":"10.3390\/molecules30061341","type":"journal-article","created":{"date-parts":[[2025,3,17]],"date-time":"2025-03-17T08:57:02Z","timestamp":1742201822000},"page":"1341","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":0,"title":["Isolation of Allosteric Tryptase Inhibitor from Methanol Extract of Rhubarb and Enhancement of Its Tryptase Inhibitory Activity by Compounds That Were Screened by In Silico Screening"],"prefix":"10.3390","volume":"30","author":[{"given":"Hidetoshi","family":"Fujii","sequence":"first","affiliation":[{"name":"Graduate School of Biostudies, Kyoto University, Kyoto 606-8502, Japan"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Moeno","family":"Ito","sequence":"additional","affiliation":[{"name":"Graduate School of Biostudies, Kyoto University, Kyoto 606-8502, Japan"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0009-0005-4864-2584","authenticated-orcid":false,"given":"Kentaro","family":"Nishioka","sequence":"additional","affiliation":[{"name":"Graduate School of Biostudies, Kyoto University, Kyoto 606-8502, Japan"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0009-0003-4790-0006","authenticated-orcid":false,"given":"Katsutoshi","family":"Nishino","sequence":"additional","affiliation":[{"name":"Graduate School of Biostudies, Kyoto University, Kyoto 606-8502, Japan"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Takanao","family":"Otsuka","sequence":"additional","affiliation":[{"name":"Department of Applied Chemistry and Biotechnology, Okayama University of Science, Okayama 700-0005, Japan"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-7109-8568","authenticated-orcid":false,"given":"Kazuhiro","family":"Irie","sequence":"additional","affiliation":[{"name":"Graduate School of Agriculture, Kyoto University, Kyoto 606-8502, Japan"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-7762-7432","authenticated-orcid":false,"given":"Takashi","family":"Tanaka","sequence":"additional","affiliation":[{"name":"Graduate School of Biomedical Sciences, Nagasaki University, Nagasaki 852-8521, Japan"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-4006-0633","authenticated-orcid":false,"given":"Masaya","family":"Nagao","sequence":"additional","affiliation":[{"name":"Graduate School of Biostudies, Kyoto University, Kyoto 606-8502, Japan"}],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"1968","published-online":{"date-parts":[[2025,3,17]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"43","DOI":"10.1006\/meth.1997.0494","article-title":"Markers of mast cell degranulation","volume":"13","author":"Hogan","year":"1997","journal-title":"Methods"},{"key":"ref_2","doi-asserted-by":"crossref","first-page":"7372","DOI":"10.1016\/S0021-9258(17)38401-6","article-title":"Regulation of tryptase from human lung mast cells by heparin. 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