The antimycotic ciclopirox olamine induces HIF‐1α stability, VEGF expression, and angiogenesis

The heterodimeric hypoxia‐inducible factor (HIF)‐1 is a master regulator of oxygen homeostasis. Protein stability and transactivation function of the α subunit are controlled by iron‐ and oxygen‐dependent hydroxylation of proline and asparagine residues. The anti‐mycotic ciclopirox olamine (CPX) is a lipophilic bidentate iron chelator that stabilizes HIF‐1α under normoxic conditions at lower concentrations than other iron chelators, probably by inhibiting HIF‐1α hydroxylation. As shown by the inhibition of iron‐dependent quenching of FITC‐labeled deferoxamine (DFX) fluorescence, CPX appears to have an even higher affinity for iron than DFX. Initial observations that treatment with 1% CPX, but not with placebo, occasionally caused reddening of wound margins in a mouse skin wound model prompted us to investigate the capability of CPX to induce angiogenesis. CPX‐induced HIF‐1‐mediated reporter gene activity and endogenous HIF‐1 target gene expression, including elevation of transcription, mRNA, and protein levels of the vascular endothelial growth factor (VEGF). In the chick chorioallantoic membrane assay, inert polymer disks containing CPX but not the solvent alone induced angiogenesis. In summary, these results suggest that CPX induces angiogenesis in vivo via HIF‐1 and VEGF induction. Therefore, CPX might serve as an alternative to recombinant VEGF treatment or to VEGF gene therapy for therapeutic angiogenesis.

[1]  D. Richardson,et al.  The role of iron in cell cycle progression and the proliferation of neoplastic cells. , 2002, Biochimica et biophysica acta.

[2]  G Zambruno,et al.  Adenovirus-mediated VEGF165 gene transfer enhances wound healing by promoting angiogenesis in CD1 diabetic mice , 2002, Gene Therapy.

[3]  R. Wenger,et al.  Cellular adaptation to hypoxia: O2‐sensing protein hydroxylases, hypoxia‐inducible transcription factors, and O2‐regulated gene expression , 2002, FASEB journal : official publication of the Federation of American Societies for Experimental Biology.

[4]  L. Zentilin,et al.  Recombinant AAV vector encoding human VEGF165 enhances wound healing , 2002, Gene Therapy.

[5]  P. Carmeliet,et al.  Conditional switching of VEGF provides new insights into adult neovascularization and pro‐angiogenic therapy , 2002, The EMBO journal.

[6]  D. Peet,et al.  Asparagine Hydroxylation of the HIF Transactivation Domain: A Hypoxic Switch , 2002, Science.

[7]  J. Isner.  Myocardial gene therapy , 2002, Nature.

[8]  S. Szabó,et al.  Gene expression and gene therapy in experimental duodenal ulceration , 2001, Journal of Physiology-Paris.

[9]  M. Gassmann,et al.  Dissecting hypoxia‐dependent and hypoxia‐independent steps in the HIF‐1α activation cascade: implications for HIF‐1α gene therapy , 2001 .

[10]  G. Koh,et al.  Gene therapy for gastric ulcers with single local injection of naked DNA encoding VEGF and angiopoietin-1. , 2001, Gastroenterology.

[11]  J. M. Arbeit,et al.  Induction of hypervascularity without leakage or inflammation in transgenic mice overexpressing hypoxia-inducible factor-1alpha. , 2001, Genes & development.

[12]  P. Ratcliffe,et al.  Independent function of two destruction domains in hypoxia‐inducible factor‐α chains activated by prolyl hydroxylation , 2001, The EMBO journal.

[13]  S. White,et al.  HIF-1α binding to VHL is regulated by stimulus-sensitive proline hydroxylation , 2001, Proceedings of the National Academy of Sciences of the United States of America.

[14]  Jin Gao,et al.  The potential of iron chelators of the pyridoxal isonicotinoyl hydrazone class as effective antiproliferative agents, IV: The mechanisms involved in inhibiting cell-cycle progression. , 2001, Blood.

[15]  M. Aoki,et al.  Gene therapy in vascular medicine: recent advances and future perspectives. , 2001, Pharmacology & therapeutics.

[16]  K. Scharffetter-Kochanek,et al.  Selective pick-up of increased iron by deferoxamine-coupled cellulose abrogates the iron-driven induction of matrix-degrading metalloproteinase 1 and lipid peroxidation in human dermal fibroblasts in vitro: a new dressing concept. , 2001, The Journal of investigative dermatology.

[17]  Michael I. Wilson,et al.  Targeting of HIF-α to the von Hippel-Lindau Ubiquitylation Complex by O2-Regulated Prolyl Hydroxylation , 2001, Science.

[18]  M. Ivan,et al.  HIFα Targeted for VHL-Mediated Destruction by Proline Hydroxylation: Implications for O2 Sensing , 2001, Science.

[19]  G. Semenza.  HIF-1 and mechanisms of hypoxia sensing. , 2001, Current opinion in cell biology.

[20]  Z. Cabantchik,et al.  Desferrioxamine-chelatable iron, a component of serum non-transferrin-bound iron, used for assessing chelation therapy. , 2001, Blood.

[21]  F. Petrat,et al.  Hypothermia injury/cold‐induced apoptosis—evidence of an increase in chelatable iron causing oxidative injury in spite of low O2−/H2O2 formation , 2000, FASEB journal : official publication of the Federation of American Societies for Experimental Biology.

[22]  M. Bohn,et al.  Dermatopharmacology of ciclopirox nail lacquer topical solution 8% in the treatment of onychomycosis. , 2000, Journal of the American Academy of Dermatology.

[23]  M. Gassmann,et al.  Epolones induce erythropoietin expression via hypoxia-inducible factor-1α activation , 2000 .

[24]  J. Ash,et al.  Lens-specific VEGF-A expression induces angioblast migration and proliferation and stimulates angiogenic remodeling. , 2000, Developmental biology.

[25]  G. Semenza,et al.  Protection from Oxidative Stress–Induced Apoptosis in Cortical Neuronal Cultures by Iron Chelators Is Associated with Enhanced DNA Binding of Hypoxia-Inducible Factor-1 and ATF-1/CREB and Increased Expression of Glycolytic Enzymes, p21waf1/cip1, and Erythropoietin , 1999, The Journal of Neuroscience.

[26]  C. Wykoff,et al.  The tumour suppressor protein VHL targets hypoxia-inducible factors for oxygen-dependent proteolysis , 1999, Nature.

[27]  M. Gassmann,et al.  Induction and nuclear translocation of hypoxia-inducible factor-1 (HIF-1): heterodimerization with ARNT is not necessary for nuclear accumulation of HIF-1alpha. , 1999, Journal of cell science.

[28]  Thomas A. Mustoe, MD, FACS,et al.  Vascular endothelial growth factor is more important than basic fibroblastic growth factor during ischemic wound healing. , 1999, Archives of surgery.

[29]  H. Pagel.  Evaluating the nephrotoxicity of cytotoxic agents using a rat kidney perfusing model , 1998 .

[30]  L. Poellinger,et al.  Signal transduction in hypoxic cells: inducible nuclear translocation and recruitment of theCBP/p300 coactivator by the hypoxia‐induciblefactor‐1α , 1998, The EMBO journal.

[31]  Jessica Lo,et al.  HIF‐1α is required for solid tumor formation and embryonic vascularization , 1998 .

[32]  M. Gassmann,et al.  Cellular and developmental control of O2 homeostasis by hypoxia-inducible factor 1 alpha. , 1998, Genes & development.

[33]  I. Orengo,et al.  Anti‐inflammatory activity of antifungal preparations , 1997, International journal of dermatology.

[34]  M. Gassmann,et al.  Oxygen-regulated Transferrin Expression Is Mediated by Hypoxia-inducible Factor-1* , 1997, The Journal of Biological Chemistry.

[35]  M. Gassmann,et al.  Functional interference between hypoxia and dioxin signal transduction pathways: competition for recruitment of the Arnt transcription factor , 1996, Molecular and cellular biology.

[36]  L. Greene,et al.  Cell cycle blockers mimosine, ciclopirox, and deferoxamine prevent the death of PC12 cells and postmitotic sympathetic neurons after removal of trophic support , 1996, The Journal of neuroscience : the official journal of the Society for Neuroscience.

[37]  M. Gassmann,et al.  Hypoxia, a Novel Inducer of Acute Phase Gene Expression in a Human Hepatoma Cell Line (*) , 1995, The Journal of Biological Chemistry.

[38]  G. Breier,et al.  Hypoxia-induced Transcriptional Activation and Increased mRNA Stability of Vascular Endothelial Growth Factor in C6 Glioma Cells (*) , 1995, The Journal of Biological Chemistry.

[39]  G. Semenza,et al.  Hypoxia-inducible factor 1 is a basic-helix-loop-helix-PAS heterodimer regulated by cellular O2 tension. , 1995, Proceedings of the National Academy of Sciences of the United States of America.

[40]  G. Semenza,et al.  Desferrioxamine induces erythropoietin gene expression and hypoxia-inducible factor 1 DNA-binding activity: implications for models of hypoxia signal transduction. , 1993, Blood.

[41]  R. Hider,et al.  Cell cycle synchronization and growth inhibition by 3-hydroxypyridin-4-one iron chelators in leukemia cell lines. , 1992, Cancer research.

[42]  K. Schulze-Osthoff,et al.  A comparative study on the effects of tumor necrosis factor‐α (TNF‐α), human angiogenic factor (h‐AF) and basic fibroblast growth factor (bFGF) on the chorioallantoic membrane of the chick embryo , 1992, The Anatomical record.

[43]  G. Coppi,et al.  HPLC method for pharmacokinetic studies on ciclopirox olamine in rabbits after intravenous and intravaginal administrations. , 1992, Farmaco.

[44]  W. Jelkmann,et al.  Isolated serum-free perfused rat kidneys release immunoreactive erythropoietin in response to hypoxia. , 1991, Endocrinology.

[45]  M. Hansen,et al.  Re-examination and further development of a precise and rapid dye method for measuring cell growth/cell kill. , 1989, Journal of immunological methods.

[46]  T. Mosmann.  Rapid colorimetric assay for cellular growth and survival: application to proliferation and cytotoxicity assays. , 1983, Journal of immunological methods.

[47]  R. Langer,et al.  Polymers for the sustained release of proteins and other macromolecules , 1976, Nature.

[48]  M. M. Bradford.  A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. , 1976, Analytical biochemistry.

[49]  H. Keberle.  The Biochemistry of Desferrioxamine and its Relation to Iron Metabolism , 1964, Annals of the New York Academy of Sciences.

[50]  D. Supp,et al.  Enhanced vascularization of cultured skin substitutes genetically modified to overexpress vascular endothelial growth factor. , 2000, The Journal of investigative dermatology.

[51]  B. Ebert,et al.  Regulation of angiogenic growth factor expression by hypoxia, transition metals, and chelating agents. , 1995, The American journal of physiology.

[52]  P. Vaupel,et al.  Therapeutic angiogenesis. , 1993, Archives of surgery.

[53]  M. Lalande,et al.  A new class of reversible cell cycle inhibitors. , 1991, Cytometry.

[54]  H. Hänel,et al.  [Therapy of seborrheic eczema with an antifungal agent with an antiphlogistic effect]. , 1991, Mycoses.

[55]  R. N. Brogden,et al.  Ciclopirox olamine 1% cream. A preliminary review of its antimicrobial activity and therapeutic use. , 1985, Drugs.

[56]  M. Freedman,et al.  Deferoxamine: a reversible S-phase inhibitor of human lymphocyte proliferation. , 1984, Blood.

[57]  W. Raether,et al.  [Microbiological laboratory studies with ciclopiroxolamine (author's transl)]. , 1981, Arzneimittel-Forschung.

[58]  E. Schütz,et al.  [Studies on the pharmacology and toxicology of ciclopiroxolamine (author's transl)]. , 1981, Arzneimittel-Forschung.

[59]  H. Yamaguchi,et al.  [Studies on the mechanism of antifungal action of ciclopiroxolamine/Inhibition of transmembrane transport of amino acid, K+ and phosphate in Candida albicans cells (author's transl)]. , 1981, Arzneimittel-Forschung.

[60]  H. Eckert,et al.  [Pharmacokinetics and biotransformation of the antimycotic drug ciclopiroxolamine in animals and man after topical and systemic administration]. , 1981, Arzneimittel-Forschung.