{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2025,11,7]],"date-time":"2025-11-07T09:20:02Z","timestamp":1762507202877,"version":"build-2065373602"},"reference-count":35,"publisher":"MDPI AG","issue":"11","license":[{"start":{"date-parts":[[2017,10,27]],"date-time":"2017-10-27T00:00:00Z","timestamp":1509062400000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Molecules"],"abstract":"<jats:p>Stratum corneum tryptic enzyme kallikrein 5 (KLK5) is a serine protease that is involved in the cell renewal and maintenance of the skin barrier function. The excessive activation of KLK5 causes an exacerbation of dermatoses, such as rosacea and atopic dermatitis. Some triterpenoids are reported to suppress the serine proteases. We aimed to investigate whether bioactive triterpenoids modulate the KLK5 protease. Nineteen triterpenoids occurring in medicinal crude drugs were evaluated using an enzymatic assay to measure the anti-KLK5 activity. The KLK5-dependent cathelicidin peptide LL-37 production in human keratinocytes was examined using immunoprecipitation and Western blotting. Screening assays for evaluating the anti-KLK5 activity revealed that ursolic acid, oleanolic acid, saikosaponin b1, tumulosic acid and pachymic acid suppressed the KLK5 protease activity, although critical molecular moieties contributing to anti-KLK5 activity were unclarified. Ursolic acid and tumulosic acid suppressed the proteolytic processing of LL-37 in keratinocytes at \u226410 \u03bcM; no cytotoxicity was observed. Both triterpenoids were detected in the plasma of rats administered orally with triterpenoid-rich crude drug Jumihaidokuto. Our study reveals that triterpenoids, such as ursolic acid and tumulosic acid, modulate the KLK5 protease activity and cathelicidin peptide production. Triterpenoids may affect the skin barrier function via the regulation of proteases.<\/jats:p>","DOI":"10.3390\/molecules22111829","type":"journal-article","created":{"date-parts":[[2017,10,27]],"date-time":"2017-10-27T11:33:28Z","timestamp":1509104008000},"page":"1829","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":16,"title":["Inhibition of Human Kallikrein 5 Protease by Triterpenoids from Natural Sources"],"prefix":"10.3390","volume":"22","author":[{"given":"Yosuke","family":"Matsubara","sequence":"first","affiliation":[{"name":"Tsumura Research Laboratories, Tsumura & Co., Ibaraki 300-1192, Japan"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-2653-5372","authenticated-orcid":false,"given":"Takashi","family":"Matsumoto","sequence":"additional","affiliation":[{"name":"Tsumura Research Laboratories, Tsumura & Co., Ibaraki 300-1192, Japan"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Junichi","family":"Koseki","sequence":"additional","affiliation":[{"name":"Tsumura Research Laboratories, Tsumura & Co., Ibaraki 300-1192, Japan"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Atsushi","family":"Kaneko","sequence":"additional","affiliation":[{"name":"Tsumura Research Laboratories, Tsumura & Co., Ibaraki 300-1192, Japan"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Setsuya","family":"Aiba","sequence":"additional","affiliation":[{"name":"Department of Dermatology, Tohoku University Graduate School of Medicine, Miyagi 980-8574, Japan"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-5495-4956","authenticated-orcid":false,"given":"Kenshi","family":"Yamasaki","sequence":"additional","affiliation":[{"name":"Department of Dermatology, Tohoku University Graduate School of Medicine, Miyagi 980-8574, Japan"}],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"1968","published-online":{"date-parts":[[2017,10,27]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"198","DOI":"10.1111\/j.0022-202X.2004.23547.x","article-title":"A proteolytic cascade of kallikreins in the stratum corneum","volume":"124","author":"Brattsand","year":"2005","journal-title":"J. Investig. Dermatol."},{"key":"ref_2","doi-asserted-by":"crossref","first-page":"1235","DOI":"10.1111\/j.0022-202X.2004.22512.x","article-title":"Degradation of corneodesmosome proteins by two serine proteases of the kallikrein family, SCTE\/KLK5\/hK5 and SCCE\/KLK7\/hK7","volume":"122","author":"Caubet","year":"2004","journal-title":"J. Investig. Dermatol."},{"key":"ref_3","doi-asserted-by":"crossref","first-page":"1173","DOI":"10.1515\/BC.2005.134","article-title":"Inhibition of human kallikreins 5 and 7 by the serine protease inhibitor lympho-epithelial Kazal-type inhibitor (LEKTI)","volume":"386","author":"Schechter","year":"2005","journal-title":"Biol. Chem."},{"key":"ref_4","doi-asserted-by":"crossref","unstructured":"Furio, L., Pampalakis, G., Michael, I.P., Nagy, A., Sotiropoulou, G., and Hovnanian, A. (2015). KLK5 inactivation reverses cutaneous hallmarks of netherton syndrome. PLoS Genet., 11.","DOI":"10.1371\/journal.pgen.1005389"},{"key":"ref_5","doi-asserted-by":"crossref","first-page":"2068","DOI":"10.1096\/fj.06-6075com","article-title":"Kallikrein-mediated proteolysis regulates the antimicrobial effects of cathelicidins in skin","volume":"20","author":"Yamasaki","year":"2006","journal-title":"FASEB J."},{"key":"ref_6","doi-asserted-by":"crossref","first-page":"975","DOI":"10.1038\/nm1616","article-title":"Increased serine protease activity and cathelicidin promotes skin inflammation in rosacea","volume":"13","author":"Yamasaki","year":"2007","journal-title":"Nat. Med."},{"key":"ref_7","doi-asserted-by":"crossref","first-page":"688","DOI":"10.1038\/jid.2010.351","article-title":"TLR2 expression is increased in rosacea and stimulates enhanced serine protease production by keratinocytes","volume":"131","author":"Yamasaki","year":"2011","journal-title":"J. Investig. Dermatol."},{"key":"ref_8","doi-asserted-by":"crossref","first-page":"12","DOI":"10.1038\/jidsymp.2011.4","article-title":"Rosacea as a disease of cathelicidins and skin innate immunity","volume":"15","author":"Yamasaki","year":"2011","journal-title":"J. Investig. Dermatol. Symp. Proc."},{"key":"ref_9","doi-asserted-by":"crossref","first-page":"513","DOI":"10.1111\/j.1600-0625.2007.00562.x","article-title":"Human tissue kallikrein expression in the stratum corneum and serum of atopic dermatitis patients","volume":"16","author":"Komatsu","year":"2007","journal-title":"Exp. Dermatol."},{"key":"ref_10","doi-asserted-by":"crossref","first-page":"1135","DOI":"10.1084\/jem.20082242","article-title":"Kallikrein 5 induces atopic dermatitis-like lesions through PAR2-mediated thymic stromal lymphopoietin expression in Netherton syndrome","volume":"206","author":"Briot","year":"2009","journal-title":"J. Exp. Med."},{"key":"ref_11","doi-asserted-by":"crossref","first-page":"1546","DOI":"10.1016\/j.biochi.2010.06.022","article-title":"Natural and synthetic inhibitors of kallikrein-related peptidases (KLKs)","volume":"92","author":"Goettig","year":"2010","journal-title":"Biochimie"},{"key":"ref_12","doi-asserted-by":"crossref","first-page":"460","DOI":"10.1016\/S0022-3565(24)36628-5","article-title":"Inhibition by boswellic acids of human leukocyte elastase","volume":"281","author":"Safayhi","year":"1997","journal-title":"J. Pharmacol. Exp. Ther."},{"key":"ref_13","doi-asserted-by":"crossref","unstructured":"Feng, L., Liu, X., Zhu, W., Guo, F., Wu, Y., Wang, R., Chen, K., Huang, C., and Li, Y. (2013). Inhibition of human neutrophil elastase by pentacyclic triterpenes. PLoS ONE, 8.","DOI":"10.1371\/journal.pone.0082794"},{"key":"ref_14","doi-asserted-by":"crossref","first-page":"599","DOI":"10.1055\/s-2001-17350","article-title":"Inhibition of trypsin and chymotrypsin by anti-inflammatory triterpenoids from Compositae flowers","volume":"67","author":"Rajic","year":"2001","journal-title":"Planta Med."},{"key":"ref_15","doi-asserted-by":"crossref","first-page":"625","DOI":"10.1111\/j.1346-8138.2007.00344.x","article-title":"Simultaneous effect of ursolic acid and oleanolic acid on epidermal permeability barrier function and epidermal keratinocyte differentiation via peroxisome proliferator-activated receptor-alpha","volume":"34","author":"Lim","year":"2007","journal-title":"J. Dermatol."},{"key":"ref_16","doi-asserted-by":"crossref","first-page":"492","DOI":"10.1248\/cpb.45.492","article-title":"Effect of the basidiomycete Poria cocos on experimental dermatitis and other inflammatory conditions","volume":"45","author":"Cuellar","year":"1997","journal-title":"Chem. Pharm. Bull."},{"key":"ref_17","doi-asserted-by":"crossref","first-page":"72","DOI":"10.1016\/j.taap.2013.03.001","article-title":"Oleanolic acid acetate inhibits atopic dermatitis and allergic contact dermatitis in a murine model","volume":"269","author":"Choi","year":"2013","journal-title":"Toxicol. Appl. Pharmacol."},{"key":"ref_18","first-page":"193","article-title":"Seijo-bofu-to, Jumi-haidoku-to and Toki-shakuyaku-san suppress rashes and incidental symptoms in acne patients","volume":"28","author":"Higaki","year":"2002","journal-title":"Drugs Exp. Clin. Res."},{"key":"ref_19","doi-asserted-by":"crossref","first-page":"4060673","DOI":"10.1155\/2016\/4060673","article-title":"Jumihaidokuto (Shi-Wei-Ba-Du-Tang), a kampo formula, decreases the disease activity of palmoplantar pustulosis","volume":"2016","author":"Mizawa","year":"2016","journal-title":"Dermatol. Res. Pract."},{"key":"ref_20","first-page":"439258","article-title":"Suppression of Propionibacterium acnes-induced dermatitis by a traditional Japanese medicine, jumihaidokuto, modifying macrophage functions","volume":"2015","author":"Sekiguchi","year":"2015","journal-title":"Evid. Based Complent. Alternat. Med."},{"key":"ref_21","doi-asserted-by":"crossref","first-page":"48","DOI":"10.1248\/bpb.22.48","article-title":"Comparison of immunopharmacological actions of 8 kinds of kampo-hozais clinically used in atopic dermatitis on delayed-type hypersensitivity in mice","volume":"22","author":"Nose","year":"1999","journal-title":"Biol. Pharm. Bull."},{"key":"ref_22","doi-asserted-by":"crossref","first-page":"521","DOI":"10.1042\/bj2770521","article-title":"Inhibition of human leucocyte elastase by ursolic acid. Evidence for a binding site for pentacyclic triterpenes","volume":"277","author":"Ying","year":"1991","journal-title":"Biochem. J."},{"key":"ref_23","doi-asserted-by":"crossref","first-page":"231","DOI":"10.1002\/biof.5520130136","article-title":"Antidiabetogenic activity of oleanolic acid glycosides from medicinal foodstuffs","volume":"13","author":"Yoshikawa","year":"2000","journal-title":"Biofactors"},{"key":"ref_24","first-page":"11","article-title":"Antimicrobial peptides in human skin disease","volume":"18","author":"Yamasaki","year":"2008","journal-title":"Eur. J. Dermatol."},{"key":"ref_25","first-page":"183","article-title":"The research on the chemical components of Schizonepeta tenuifolia Briq","volume":"24","author":"Zhang","year":"2001","journal-title":"J. Chin. Med. Mater."},{"key":"ref_26","doi-asserted-by":"crossref","first-page":"681","DOI":"10.1055\/s-0030-1270823","article-title":"Chemical constituents and pharmacological properties of Poria cocos","volume":"77","author":"Rios","year":"2011","journal-title":"Planta Med."},{"key":"ref_27","doi-asserted-by":"crossref","first-page":"1131","DOI":"10.1002\/ptr.4833","article-title":"Schizonepeta tenuifolia inhibits the development of atopic dermatitis in mice","volume":"27","author":"Choi","year":"2013","journal-title":"Phytother. Res."},{"key":"ref_28","doi-asserted-by":"crossref","first-page":"599","DOI":"10.1248\/bpb.23.599","article-title":"Inhibitory effects of methanol extracts of herbal medicines on substance P-induced itch-scratch response","volume":"23","author":"Tohda","year":"2000","journal-title":"Biol. Pharm. Bull."},{"key":"ref_29","doi-asserted-by":"crossref","first-page":"30","DOI":"10.1177\/0091270002042001003","article-title":"Schizonepeta tenuifolia: Chemistry, pharmacology, and clinical applications","volume":"42","author":"Fung","year":"2002","journal-title":"J. Clin. Pharmacol."},{"key":"ref_30","doi-asserted-by":"crossref","first-page":"847","DOI":"10.1248\/cpb.44.847","article-title":"Isolation of inhibitors of TPA-induced mouse ear edema from Hoelen, Poria cocos","volume":"44","author":"Nukaya","year":"1996","journal-title":"Chem. Pharm. Bull."},{"key":"ref_31","doi-asserted-by":"crossref","first-page":"2877","DOI":"10.1007\/s00216-011-4651-x","article-title":"Development of a liquid chromatography-mass spectrometry method for the determination of ursolic acid in rat plasma and tissue: Application to the pharmacokinetic and tissue distribution study","volume":"399","author":"Chen","year":"2011","journal-title":"Anal. Bioanal. Chem."},{"key":"ref_32","doi-asserted-by":"crossref","first-page":"51","DOI":"10.1002\/bdd.530","article-title":"Dose-linear pharmacokinetics of oleanolic acid after intravenous and oral administration in rats","volume":"28","author":"Jeong","year":"2007","journal-title":"Biopharm. Drug Dispos."},{"key":"ref_33","doi-asserted-by":"crossref","unstructured":"Matsubara, Y., Matsumoto, T., Sekiguchi, K., Koseki, J., Kaneko, A., Yamaguchi, T., Kurihara, Y., and Kobayashi, H. (2017). Oral administration of the Japanese traditional medicine keishibukuryogan-ka-yokuinin decreases reactive oxygen metabolites in rat plasma: Identification of chemical constituents contributing to antioxidant activity. Molecules, 22.","DOI":"10.3390\/molecules22020256"},{"key":"ref_34","doi-asserted-by":"crossref","first-page":"904","DOI":"10.1002\/jms.3230","article-title":"A UFLC-MS\/MS method with a switching ionization mode for simultaneous quantitation of polygalaxanthone III, four ginsenosides and tumulosic acid in rat plasma: Application to a comparative pharmacokinetic study in normal and Alzheimer\u2019s disease rats","volume":"48","author":"Lv","year":"2013","journal-title":"J. Mass Spectrom."},{"key":"ref_35","doi-asserted-by":"crossref","first-page":"117","DOI":"10.1016\/j.fitote.2011.10.001","article-title":"Comparative pharmacokinetics of three triterpene acids in rat plasma after oral administration of Poria extract and its formulated herbal preparation: GuiZhi-FuLing capsule","volume":"83","author":"Xiao","year":"2012","journal-title":"Fitoterapia"}],"container-title":["Molecules"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/1420-3049\/22\/11\/1829\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,10,11]],"date-time":"2025-10-11T18:48:41Z","timestamp":1760208521000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/1420-3049\/22\/11\/1829"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2017,10,27]]},"references-count":35,"journal-issue":{"issue":"11","published-online":{"date-parts":[[2017,11]]}},"alternative-id":["molecules22111829"],"URL":"https:\/\/doi.org\/10.3390\/molecules22111829","relation":{},"ISSN":["1420-3049"],"issn-type":[{"type":"electronic","value":"1420-3049"}],"subject":[],"published":{"date-parts":[[2017,10,27]]}}}