title: Synthesis and Functional Characterization of Novel Sialyl LewisX Mimic-Decorated Liposomes for E-selectin-Mediated Targeting to Inflamed Endothelial Cells.
abstract: Sialyl LewisX (sLeX) is a natural ligand of E-selectin that is overexpressed by inflamed and tumor endothelium. Although sLeX is a potential ligand for drug targeting, synthesis of the tetrasaccharide is complicated with many reaction steps. In this study, structurally simplified novel sLeX analogues were designed and linked with 1,2-distearoyl-sn-glycero-3-phosphoethanolamine-polyethylene glycol-2000 (DSPE-PEG) for E-selectin-mediated liposomal delivery. The sLeX structural simplification strategies include (1) replacement of the Gal-GlcNAc disaccharide unit with lactose to reduce many initial steps and (2) substitution of neuraminic acid with a negatively charged group, i.e., 3'-sulfo, 3'-carboxymethyl (3'-CM), or 3'-(1-carboxy)ethyl (3'-CE). While all the liposomes developed were similar in particle size and charge, the 3'-CE sLeX mimic liposome demonstrated the highest uptake in inflammatory cytokine-treated human umbilical vein endothelial cells (HUVECs), being even more potent than native sLeX-decorated liposomes. Inhibition studies using antiselectin antibodies revealed that their uptake was mediated primarily by overexpressed E-selectin on inflamed HUVECs. Molecular dynamics simulations were performed to gain mechanistic insight into the E-selectin binding differences among native and mimic sLeX. The terminally branched methyl group of the 3'-CE sLeX mimic oriented and faced the bulk hydrophilic solution during E-selectin binding. Since this state is entropically unfavorable, the 3'-CE sLeX mimic molecule might be pushed toward the binding pocket of E-selectin by a hydrophobic effect, leading to a higher probability of hydrogen-bond formation than native sLeX and the 3'-CM sLeX mimic. This corresponded with the fact that the 3'-CE sLeX mimic liposome exhibited much greater uptake than the 3'-CM sLeX mimic liposome.
authors.affiliation: Department of Drug Delivery Research, Graduate School of Pharmaceutical Sciences, Kyoto University , 46-29 Yoshidashimoadachi-cho, Sakyo-ku, Kyoto 606-8302, Japan.
authors.forename: Chanikarn
authors.initials: C
authors.lastname: Chantarasrivong
authors.affiliation: Department of Applied Bioorganic Chemistry, Gifu University , 1-1 Yanagido, Gifu-shi, Gifu 501-1193, Japan.|Institute for Integrated Cell-Material Sciences (WPI-iCeMS), Kyoto University , Yoshidaushinomiya-cho, Sakyo-ku, Kyoto 606-8501, Japan.
authors.forename: Akiharu
authors.initials: A
authors.lastname: Ueki
authors.affiliation: Department of Applied Bioorganic Chemistry, Gifu University , 1-1 Yanagido, Gifu-shi, Gifu 501-1193, Japan.
authors.forename: Ryutaro
authors.initials: R
authors.lastname: Ohyama
authors.affiliation: Department of Drug Delivery Research, Graduate School of Pharmaceutical Sciences, Kyoto University , 46-29 Yoshidashimoadachi-cho, Sakyo-ku, Kyoto 606-8302, Japan.
authors.forename: Johan
authors.initials: J
authors.lastname: Unga
authors.affiliation: Department of Pharmaceutical Sciences, Faculty of Pharmacy, Kindai University , 3-4-1 Kowakae, Higashi-Osaka, Osaka 577-8502, Japan.
authors.forename: Shinya
authors.initials: S
authors.lastname: Nakamura
authors.affiliation: Department of Pharmaceutical Sciences, Faculty of Pharmacy, Kindai University , 3-4-1 Kowakae, Higashi-Osaka, Osaka 577-8502, Japan.
authors.forename: Isao
authors.initials: I
authors.lastname: Nakanishi
authors.affiliation: Department of Drug Delivery Research, Graduate School of Pharmaceutical Sciences, Kyoto University , 46-29 Yoshidashimoadachi-cho, Sakyo-ku, Kyoto 606-8302, Japan.
authors.forename: Yuriko
authors.initials: Y
authors.lastname: Higuchi
authors.affiliation: Department of Drug Delivery Research, Graduate School of Pharmaceutical Sciences, Kyoto University , 46-29 Yoshidashimoadachi-cho, Sakyo-ku, Kyoto 606-8302, Japan.
authors.forename: Shigeru
authors.identifier: 0000-0001-8765-2463
authors.initials: S
authors.lastname: Kawakami
authors.affiliation: Department of Applied Bioorganic Chemistry, Gifu University , 1-1 Yanagido, Gifu-shi, Gifu 501-1193, Japan.|Institute for Integrated Cell-Material Sciences (WPI-iCeMS), Kyoto University , Yoshidaushinomiya-cho, Sakyo-ku, Kyoto 606-8501, Japan.|Gifu Center for Highly Advanced Integration of Nano and Life Sciences (G-CHAIN), Gifu University , 1-1 Yanagido, Gifu-shi, Gifu 501-1193, Japan.
authors.forename: Hiromune
authors.initials: H
authors.lastname: Ando
authors.affiliation: Department of Applied Bioorganic Chemistry, Gifu University , 1-1 Yanagido, Gifu-shi, Gifu 501-1193, Japan.
authors.forename: Akihiro
authors.initials: A
authors.lastname: Imamura
authors.affiliation: Department of Applied Bioorganic Chemistry, Gifu University , 1-1 Yanagido, Gifu-shi, Gifu 501-1193, Japan.|Gifu Center for Highly Advanced Integration of Nano and Life Sciences (G-CHAIN), Gifu University , 1-1 Yanagido, Gifu-shi, Gifu 501-1193, Japan.
authors.forename: Hideharu
authors.initials: H
authors.lastname: Ishida
authors.affiliation: Department of Drug Delivery Research, Graduate School of Pharmaceutical Sciences, Kyoto University , 46-29 Yoshidashimoadachi-cho, Sakyo-ku, Kyoto 606-8302, Japan.
authors.forename: Fumiyoshi
authors.identifier: 0000-0002-3503-8696
authors.initials: F
authors.lastname: Yamashita
authors.affiliation: Department of Applied Bioorganic Chemistry, Gifu University , 1-1 Yanagido, Gifu-shi, Gifu 501-1193, Japan.|Institute for Integrated Cell-Material Sciences (WPI-iCeMS), Kyoto University , Yoshidaushinomiya-cho, Sakyo-ku, Kyoto 606-8501, Japan.
authors.forename: Makoto
authors.initials: M
authors.lastname: Kiso
authors.affiliation: Department of Drug Delivery Research, Graduate School of Pharmaceutical Sciences, Kyoto University , 46-29 Yoshidashimoadachi-cho, Sakyo-ku, Kyoto 606-8302, Japan.|Institute for Integrated Cell-Material Sciences (WPI-iCeMS), Kyoto University , Yoshidaushinomiya-cho, Sakyo-ku, Kyoto 606-8501, Japan.
authors.forename: Mitsuru
authors.initials: M
authors.lastname: Hashida
mesh_terms: D019040:E-Selectin / Q000737:chemistry*; D042783:Endothelial Cells / Q000378:metabolism*; D061307:Human Umbilical Vein Endothelial Cells; D006801:Humans; D008081:Liposomes / Q000737:chemistry* / Q000378:metabolism; D056004:Molecular Dynamics Simulation
chemical_list: D019040:E-Selectin; D008081:Liposomes
keywords: E-selectin; in vitro uptake; inflamed endothelial cell; molecular dynamics simulation; sialyl LewisX mimic; targeted liposomal delivery