title: Effect of nitroxoline on angiogenesis and growth of human bladder cancer.
abstract: BACKGROUND
  Angiogenesis plays an important role in tumor growth and metastasis; therefore, inhibition of angiogenesis is a promising strategy for developing new anticancer drugs. Type 2 methionine aminopeptidase (MetAP2) protein is likely a molecular target of angiogenesis inhibitors.
  
  
  METHODS
  Nitroxoline, an antibiotic used to treat urinary tract infections, was identified from a high-throughput screen of a library of 175,000 compounds for MetAP2 inhibitors and from a parallel screen using the Johns Hopkins Drug Library to identify currently used clinical drugs that can also inhibit human umbilical vein endothelial cells (HUVEC) proliferation. To investigate the mechanism of action of nitroxoline, inhibition of MetAP2 activity and induction of senescence were assessed in HUVEC. To test the antiangiogenic activity of nitroxoline, endothelial tube formation in Matrigel and microvessel formation in Matrigel plugs in vivo were assessed. Antitumor efficacy of nitroxoline was evaluated in mouse models of human breast cancer xenograft (n = 10) and bladder cancer orthotopic xenograft (n = 11). Furthermore, the mechanism of action of nitroxoline was investigated in vivo.
  
  
  RESULTS
  Nitroxoline inhibited MetAP2 activity in vitro (half maximal inhibitory concentration [IC(50)] = 54.8 nM, 95% confidence interval [CI] = 22.6 to 132.8 nM) and HUVEC proliferation (IC(50) = 1.9 μM, 95% CI = 1.54 to 2.39 μM). Nitroxoline inhibited MetAP2 activity in HUVEC in a dose-dependent manner and induced premature senescence in a biphasic manner. Nitroxoline inhibited endothelial tube formation in Matrigel and reduced microvessel density in vivo. Mice (five per group) treated with nitroxoline showed a 60% reduction in tumor volume in breast cancer xenografts (tumor volume on day 30, vehicle vs nitroxoline, mean = 215.4 vs 86.5 mm(3), difference = 128.9 mm(3), 95% CI = 32.9 to 225.0 mm(3), P = .012) and statistically significantly inhibited growth of bladder cancer in an orthotopic mouse model (tumor bioluminescence intensities of vehicle [n = 5] vs nitroxoline [n = 6], P = .045).
  
  
  CONCLUSIONS
  Nitroxoline shows promise as a potential therapeutic antiangiogenic agent.
authors.affiliation: Department of Pharmacology and Molecular Sciences, Johns Hopkins School of Medicine, Baltimore, MD 21205, USA.
authors.forename: Joong Sup
authors.initials: JS
authors.lastname: Shim
authors.forename: Yoshiyuki
authors.initials: Y
authors.lastname: Matsui
authors.forename: Shridhar
authors.initials: S
authors.lastname: Bhat
authors.forename: Benjamin A
authors.initials: BA
authors.lastname: Nacev
authors.forename: Jing
authors.initials: J
authors.lastname: Xu
authors.forename: Hyo-eun C
authors.initials: HE
authors.lastname: Bhang
authors.forename: Surajit
authors.initials: S
authors.lastname: Dhara
authors.forename: Kee Chung
authors.initials: KC
authors.lastname: Han
authors.forename: Curtis R
authors.initials: CR
authors.lastname: Chong
authors.forename: Martin G
authors.initials: MG
authors.lastname: Pomper
authors.forename: Alan
authors.initials: A
authors.lastname: So
authors.forename: Jun O
authors.initials: JO
authors.lastname: Liu
mesh_terms: D000107:Acetylation / Q000187:drug effects; D000626:Aminopeptidases / Q000037:antagonists & inhibitors* / Q000378:metabolism; D020533:Angiogenesis Inhibitors / Q000494:pharmacology*; D000818:Animals; D000892:Anti-Infective Agents, Urinary / Q000494:pharmacology; D001943:Breast Neoplasms / Q000098:blood supply* / Q000188:drug therapy*; D049109:Cell Proliferation / Q000187:drug effects; D003094:Collagen; D003510:Cyclohexanes / Q000494:pharmacology; D004195:Disease Models, Animal; D004338:Drug Combinations; D004357:Drug Synergism; D042783:Endothelial Cells / Q000187:drug effects / Q000378:metabolism / Q000473:pathology; D004730:Endothelium, Vascular / Q000187:drug effects / Q000378:metabolism / Q000473:pathology; D005260:Female; D006023:Glycoproteins / Q000037:antagonists & inhibitors* / Q000378:metabolism; D006801:Humans; D015151:Immunoblotting; D007150:Immunohistochemistry; D007797:Laminin; D063208:Methionyl Aminopeptidases; D051379:Mice; D008819:Mice, Nude; D009389:Neovascularization, Pathologic / Q000517:prevention & control*; D009600:Nitroquinolines / Q000494:pharmacology*; D000077609:O-(Chloroacetylcarbamoyl)fumagillol; D011509:Proteoglycans; D034741:RNA, Small Interfering; D011994:Recombinant Proteins; D020133:Reverse Transcriptase Polymerase Chain Reaction; D012717:Sesquiterpenes / Q000494:pharmacology; D056564:Sirtuin 1 / Q000037:antagonists & inhibitors* / Q000378:metabolism; D014162:Transfection; D014471:Umbilical Veins / Q000378:metabolism / Q000473:pathology; D001749:Urinary Bladder Neoplasms / Q000098:blood supply* / Q000188:drug therapy*; D023041:Xenograft Model Antitumor Assays
chemical_list: D020533:Angiogenesis Inhibitors; D000892:Anti-Infective Agents, Urinary; D003510:Cyclohexanes; D004338:Drug Combinations; D006023:Glycoproteins; D007797:Laminin; D009600:Nitroquinolines; D011509:Proteoglycans; D034741:RNA, Small Interfering; D011994:Recombinant Proteins; D012717:Sesquiterpenes; C059485:matrigel; D003094:Collagen; C005308:nitroxoline; D000626:Aminopeptidases; C081311:METAP2 protein, human; D063208:Methionyl Aminopeptidases; C447939:SIRT1 protein, human; D056564:Sirtuin 1; D000077609:O-(Chloroacetylcarbamoyl)fumagillol