title: Claudin-1 Binder Enhances Epidermal Permeability in a Human Keratinocyte Model.
abstract: Tight junctions (TJs) are complex biochemical structures that seal the intercellular space and prevent the free movement of solutes across epithelial cell sheets. Modulating the TJ seal is a promising option for increasing the transdermal absorption of drugs. Within TJs, the binding of the claudin (CLDN) family of tetratransmembrane proteins through cis- and trans-interactions is an integral part of seal formation. Because epidermal TJs contain CLDN-1 and CLDN-4, a binder for these CLDNs may be a useful modulator of the permeability of the epidermal barrier. Here, we investigated whether m19, which can bind to CLDN-1/-4 (also CLDN-2/-5), modulates the integrity of epidermal TJs and the permeability of cell sheets to solutes. Treatment of normal human epidermal keratinocytes (NHEKs) with the CLDN binder reduced the integrity of TJs. A CLDN-1-specific binder (a monoclonal antibody, clone 7A5) also weakened the TJ seal in NHEKs. Although m19 attenuated the TJ barrier in human intestinal epithelial cells (Caco-2), 7A5 did not. Treatment of NHEKs with 7A5 enhanced permeation of a paracellular permeation marker. These findings indicate that CLDN-1 is a potential target for modulating the permeability of the epidermis, and that our CLDN-1 binder is a promising candidate molecule for development as a dermal absorption enhancer.
authors.affiliation: Laboratory of Bio-Functional Molecular Chemistry, Graduate School of Pharmaceutical Sciences, Osaka University, Osaka, Japan (M.N., S.N., M.I., M.Y., A.W., K.Y., M.K.); Asubio Pharma Co., Ltd., Hyogo, Japan (S.T.); Department of Biochemistry and Cell Biology, National Institute of Infectious Diseases, Tokyo, Japan (Y.S., M.F.); and Proteo-Science Center, Ehime University, Ehime, Japan (H.T., T.S.).
authors.forename: Misaki
authors.initials: M
authors.lastname: Nakajima
authors.affiliation: Laboratory of Bio-Functional Molecular Chemistry, Graduate School of Pharmaceutical Sciences, Osaka University, Osaka, Japan (M.N., S.N., M.I., M.Y., A.W., K.Y., M.K.); Asubio Pharma Co., Ltd., Hyogo, Japan (S.T.); Department of Biochemistry and Cell Biology, National Institute of Infectious Diseases, Tokyo, Japan (Y.S., M.F.); and Proteo-Science Center, Ehime University, Ehime, Japan (H.T., T.S.).
authors.forename: Shotaro
authors.initials: S
authors.lastname: Nagase
authors.affiliation: Laboratory of Bio-Functional Molecular Chemistry, Graduate School of Pharmaceutical Sciences, Osaka University, Osaka, Japan (M.N., S.N., M.I., M.Y., A.W., K.Y., M.K.); Asubio Pharma Co., Ltd., Hyogo, Japan (S.T.); Department of Biochemistry and Cell Biology, National Institute of Infectious Diseases, Tokyo, Japan (Y.S., M.F.); and Proteo-Science Center, Ehime University, Ehime, Japan (H.T., T.S.).
authors.forename: Manami
authors.initials: M
authors.lastname: Iida
authors.affiliation: Laboratory of Bio-Functional Molecular Chemistry, Graduate School of Pharmaceutical Sciences, Osaka University, Osaka, Japan (M.N., S.N., M.I., M.Y., A.W., K.Y., M.K.); Asubio Pharma Co., Ltd., Hyogo, Japan (S.T.); Department of Biochemistry and Cell Biology, National Institute of Infectious Diseases, Tokyo, Japan (Y.S., M.F.); and Proteo-Science Center, Ehime University, Ehime, Japan (H.T., T.S.).
authors.forename: Shuji
authors.initials: S
authors.lastname: Takeda
authors.affiliation: Laboratory of Bio-Functional Molecular Chemistry, Graduate School of Pharmaceutical Sciences, Osaka University, Osaka, Japan (M.N., S.N., M.I., M.Y., A.W., K.Y., M.K.); Asubio Pharma Co., Ltd., Hyogo, Japan (S.T.); Department of Biochemistry and Cell Biology, National Institute of Infectious Diseases, Tokyo, Japan (Y.S., M.F.); and Proteo-Science Center, Ehime University, Ehime, Japan (H.T., T.S.).
authors.forename: Mayo
authors.initials: M
authors.lastname: Yamashita
authors.affiliation: Laboratory of Bio-Functional Molecular Chemistry, Graduate School of Pharmaceutical Sciences, Osaka University, Osaka, Japan (M.N., S.N., M.I., M.Y., A.W., K.Y., M.K.); Asubio Pharma Co., Ltd., Hyogo, Japan (S.T.); Department of Biochemistry and Cell Biology, National Institute of Infectious Diseases, Tokyo, Japan (Y.S., M.F.); and Proteo-Science Center, Ehime University, Ehime, Japan (H.T., T.S.).
authors.forename: Akihiro
authors.initials: A
authors.lastname: Watari
authors.affiliation: Laboratory of Bio-Functional Molecular Chemistry, Graduate School of Pharmaceutical Sciences, Osaka University, Osaka, Japan (M.N., S.N., M.I., M.Y., A.W., K.Y., M.K.); Asubio Pharma Co., Ltd., Hyogo, Japan (S.T.); Department of Biochemistry and Cell Biology, National Institute of Infectious Diseases, Tokyo, Japan (Y.S., M.F.); and Proteo-Science Center, Ehime University, Ehime, Japan (H.T., T.S.).
authors.forename: Yoshitaka
authors.initials: Y
authors.lastname: Shirasago
authors.affiliation: Laboratory of Bio-Functional Molecular Chemistry, Graduate School of Pharmaceutical Sciences, Osaka University, Osaka, Japan (M.N., S.N., M.I., M.Y., A.W., K.Y., M.K.); Asubio Pharma Co., Ltd., Hyogo, Japan (S.T.); Department of Biochemistry and Cell Biology, National Institute of Infectious Diseases, Tokyo, Japan (Y.S., M.F.); and Proteo-Science Center, Ehime University, Ehime, Japan (H.T., T.S.).
authors.forename: Masayoshi
authors.initials: M
authors.lastname: Fukasawa
authors.affiliation: Laboratory of Bio-Functional Molecular Chemistry, Graduate School of Pharmaceutical Sciences, Osaka University, Osaka, Japan (M.N., S.N., M.I., M.Y., A.W., K.Y., M.K.); Asubio Pharma Co., Ltd., Hyogo, Japan (S.T.); Department of Biochemistry and Cell Biology, National Institute of Infectious Diseases, Tokyo, Japan (Y.S., M.F.); and Proteo-Science Center, Ehime University, Ehime, Japan (H.T., T.S.).
authors.forename: Hiroyuki
authors.initials: H
authors.lastname: Takeda
authors.affiliation: Laboratory of Bio-Functional Molecular Chemistry, Graduate School of Pharmaceutical Sciences, Osaka University, Osaka, Japan (M.N., S.N., M.I., M.Y., A.W., K.Y., M.K.); Asubio Pharma Co., Ltd., Hyogo, Japan (S.T.); Department of Biochemistry and Cell Biology, National Institute of Infectious Diseases, Tokyo, Japan (Y.S., M.F.); and Proteo-Science Center, Ehime University, Ehime, Japan (H.T., T.S.).
authors.forename: Tatsuya
authors.initials: T
authors.lastname: Sawasaki
authors.affiliation: Laboratory of Bio-Functional Molecular Chemistry, Graduate School of Pharmaceutical Sciences, Osaka University, Osaka, Japan (M.N., S.N., M.I., M.Y., A.W., K.Y., M.K.); Asubio Pharma Co., Ltd., Hyogo, Japan (S.T.); Department of Biochemistry and Cell Biology, National Institute of Infectious Diseases, Tokyo, Japan (Y.S., M.F.); and Proteo-Science Center, Ehime University, Ehime, Japan (H.T., T.S.).
authors.forename: Kiyohito
authors.initials: K
authors.lastname: Yagi
authors.affiliation: Laboratory of Bio-Functional Molecular Chemistry, Graduate School of Pharmaceutical Sciences, Osaka University, Osaka, Japan (M.N., S.N., M.I., M.Y., A.W., K.Y., M.K.); Asubio Pharma Co., Ltd., Hyogo, Japan (S.T.); Department of Biochemistry and Cell Biology, National Institute of Infectious Diseases, Tokyo, Japan (Y.S., M.F.); and Proteo-Science Center, Ehime University, Ehime, Japan (H.T., T.S.) masuo@phs.osaka-u.ac.jp.
authors.forename: Masuo
authors.initials: M
authors.lastname: Kondoh
mesh_terms: D018938:Caco-2 Cells; D045744:Cell Line, Tumor; D062445:Claudin-1 / Q000378:metabolism*; D062506:Claudin-4 / Q000378:metabolism; D004817:Epidermis / Q000378:metabolism; D004847:Epithelial Cells / Q000378:metabolism; D006801:Humans; D015603:Keratinocytes / Q000378:metabolism*; D008565:Membrane Proteins / Q000378:metabolism; D010539:Permeability; D011485:Protein Binding / Q000502:physiology*; D019108:Tight Junctions / Q000378:metabolism
chemical_list: D062445:Claudin-1; D062506:Claudin-4; D008565:Membrane Proteins