title: Water soluble PEG-conjugate of xanthine oxidase inhibitor, PEG-AHPP micelles, as a novel therapeutic for ROS related inflammatory bowel diseases.
abstract: Xanthine oxidase (XO) is one of the major enzymes to generate superoxide anion (O2(-)), that is frequently associated with various diseases involving reactive oxygen species (ROS). 4-Amino-6-hydroxypyrazolo[3,4-d]pyrimidine (AHPP) is a potent XO inhibitor showing therapeutic potential for oxidative inflammatory diseases. However its very poor aqueous solubility makes pharmaceutical application difficult. To overcome this drawback, we have successfully synthesized a water soluble polyethylene glycol (PEG) conjugate of AHPP (PEG-AHPP) that exhibited good water solubility, forming micelles in aqueous solution. In the present study, the in vivo pharmacokinetics of this PEG-AHPP was examined. Further its therapeutic potential was investigated in dextran sulfate sodium (DSS) induced mouse colitis model. Compared to parental AHPP, the plasma t1/2 of PEG-AHPP was increased remarkably from 3h to 14h, indicating macromolecular nature of AHPP in circulation. In the DSS induced colitis model, oral administration of 2% DSS in drinking water resulted in the progression of the colitis with diarrhea and hematochezia as well as shortening of the large bowel. Administration of PEG-AHPP intravenously (10mg/kg) or orally (20mg/kg) suppressed pathogenesis significantly; namely diarrhea was reduced markedly, and the length of large bowel returned to almost normal level. Pathological examination clearly revealed improvement of colonic ulcer or necrosis. Production of inflammatory cytokines, i.e., interleukin-6 and tumor necrosis factor (TNF)-α, was significantly increased in DSS-induced colitis mice. However, it was markedly suppressed by PEG-AHPP administration. Similar results were found when serum 8-hydroxydeoxyguanosine (8-OHdG) and thiobarbituric acid reactive substances (TBARS), that are the index of oxidative injury, were measured. PEG-AHPP thus may be a potential candidate drug for ROS-related diseases including inflammatory bowel disease.
authors.affiliation: DDS Research Institute, Sojo University, Ikeda 4-22-1, Nishi-ku, Kumamoto 860-0082, Japan; Laboratory of Microbiology & Oncology, Faculty of Pharmaceutical Sciences, Sojo University, Japan; School of Public Health, Anhui Medical University, 81th Meishan Road, Hefei City, Anhui Province 230032, China. Electronic address: fangjun@ph.sojo-u.ac.jp.
authors.forename: Jun
authors.initials: J
authors.lastname: Fang
authors.affiliation: DDS Research Institute, Sojo University, Ikeda 4-22-1, Nishi-ku, Kumamoto 860-0082, Japan; Department of General Surgery, Sheng Jing Hospital, China Medical University, Shenyang City, Liaoning Province 110004, China.
authors.forename: Hongzhuan
authors.initials: H
authors.lastname: Yin
authors.affiliation: DDS Research Institute, Sojo University, Ikeda 4-22-1, Nishi-ku, Kumamoto 860-0082, Japan; Department of Applied Microbial Technology, Faculty of Biotechnology & Life Science, Sojo University, Japan.
authors.forename: Long
authors.initials: L
authors.lastname: Liao
authors.affiliation: DDS Research Institute, Sojo University, Ikeda 4-22-1, Nishi-ku, Kumamoto 860-0082, Japan; Department of Applied Microbial Technology, Faculty of Biotechnology & Life Science, Sojo University, Japan.
authors.forename: Haibo
authors.initials: H
authors.lastname: Qin
authors.affiliation: DDS Research Institute, Sojo University, Ikeda 4-22-1, Nishi-ku, Kumamoto 860-0082, Japan; Laboratory of Microbiology & Oncology, Faculty of Pharmaceutical Sciences, Sojo University, Japan.
authors.forename: Fumiko
authors.initials: F
authors.lastname: Ueda
authors.affiliation: DDS Research Institute, Sojo University, Ikeda 4-22-1, Nishi-ku, Kumamoto 860-0082, Japan; Laboratory of Microbiology & Oncology, Faculty of Pharmaceutical Sciences, Sojo University, Japan.
authors.forename: Kyoko
authors.initials: K
authors.lastname: Uemura
authors.affiliation: DDS Research Institute, Sojo University, Ikeda 4-22-1, Nishi-ku, Kumamoto 860-0082, Japan; Laboratory of Microbiology & Oncology, Faculty of Pharmaceutical Sciences, Sojo University, Japan.
authors.forename: Kanami
authors.initials: K
authors.lastname: Eguchi
authors.affiliation: DDS Research Institute, Sojo University, Ikeda 4-22-1, Nishi-ku, Kumamoto 860-0082, Japan; Department of Nanoscience and Applied Chemistry, Graduate School of Engineering, Sojo University, Japan.
authors.forename: Gahininath Y
authors.initials: GY
authors.lastname: Bharate
authors.affiliation: DDS Research Institute, Sojo University, Ikeda 4-22-1, Nishi-ku, Kumamoto 860-0082, Japan; Laboratory of Microbiology & Oncology, Faculty of Pharmaceutical Sciences, Sojo University, Japan. Electronic address: hirmaeda@ph.sojo-u.ac.jp.
authors.forename: Hiroshi
authors.initials: H
authors.lastname: Maeda
mesh_terms: D000080242:8-Hydroxy-2'-Deoxyguanosine; D000818:Animals; D018938:Caco-2 Cells; D003092:Colitis / Q000139:chemically induced / Q000188:drug therapy* / Q000276:immunology / Q000378:metabolism; D003106:Colon / Q000187:drug effects / Q000276:immunology / Q000378:metabolism / Q000473:pathology; D016207:Cytokines / Q000276:immunology; D003849:Deoxyguanosine / Q000031:analogs & derivatives / Q000097:blood; D016264:Dextran Sulfate; D005260:Female; D006801:Humans; D008813:Mice, Inbred ICR; D008823:Micelles; D010117:Oxypurinol / Q000008:administration & dosage / Q000031:analogs & derivatives* / Q000493:pharmacokinetics / Q000627:therapeutic use; D011092:Polyethylene Glycols / Q000008:administration & dosage* / Q000493:pharmacokinetics / Q000627:therapeutic use; D017382:Reactive Oxygen Species; D012995:Solubility; D017392:Thiobarbituric Acid Reactive Substances / Q000032:analysis; D014867:Water / Q000737:chemistry; D014969:Xanthine Oxidase / Q000037:antagonists & inhibitors* / Q000097:blood / Q000378:metabolism
chemical_list: C548914:4-amino-6-hydroxypyrazolo(3,4-d)pyrimidine; D016207:Cytokines; D008823:Micelles; D017382:Reactive Oxygen Species; D017392:Thiobarbituric Acid Reactive Substances; D014867:Water; D011092:Polyethylene Glycols; D000080242:8-Hydroxy-2'-Deoxyguanosine; D016264:Dextran Sulfate; D014969:Xanthine Oxidase; D003849:Deoxyguanosine; D010117:Oxypurinol
keywords: AHPP; Inflammation; Inflammatory bowel disease; Micelle drug; Reactive oxygen species; Xanthine oxidase inhibitor