Efficacy of fumagillin bicyclohexylamine on experimental corneal neovascularization in rat model

  • M. Fatih Aşula
  • I. Umut OnurEmail author
  • F. Ulviye Yigit
Original Paper



Fumagillin has been previously used to treat corneal microsporidial keratitis and also identified as an angiogenesis inhibitor. This study aimed to evaluate efficacy of fumagillin bicyclohexylamine on the rat model of corneal neovascularization induced by silver nitrate cauterization.


Twenty-four Albino Wistar rats (n = 24) were divided into three groups. Following silver nitrate-induced corneal injury, eyes in Group 1 received one drop of 5 mg/mL topical fumagillin bicyclohexylamine four times daily for 10 days. Group 2 received subconjunctival injection of 0.1 mL fumagillin bicyclohexylamine (2.5 mg/mL) on day 1 and day 5. Group 3 received artificial tears and lubricants four times daily for 10 days as control. On day 10, animals were sacrificed. Corneal specimens were obtained and prepared to assess vascular endothelial growth factor (VEGF-C) levels and corneal angiogenic microvessel density.


There was no significant difference in VEGF-C levels between the groups (P = 0.994). Assessment of angiogenic microvessel density for peripheral corneal zone also did not reveal significant difference between the groups (P = 0.113). However, mean vascular density in Group 1 and Group 2 was significantly higher for both midperipheral and central corneal zones in comparison with Group 3 (P = 0.003, P = 0.015).


Previously proved to be effective for treatment of microsporidial keratitis in humans, topical and subconjunctival concentration or dosing of fumagillin bicyclohexylamine failed to reduce corneal neovascularization induced by silver nitrate in this study. Further studies comparing different concentrations and dosing may detect inhibitory effects of fumagillin on corneal neovascularization without inducing toxicity.


Fumagillin bicyclohexylamine Corneal neovascularization VEGF-C Silver nitrate Rat cornea 



Authors thank to Murat Icen, MD for English proofreading.


This research did not receive any specific grant from funding agencies in the public, commercial or not-for-profit sectors.

Compliance with ethical standards

Conflict of interest

The authors declare that they have no conflict of interest.

Animal rights statement

All procedures performed in studies involving animals were in accordance with the ethical standards of the institution or practice at which the studies were conducted.


  1. 1.
    Joussen AM, Beecken WD, Moromizato Y, Schwartz A, Kirchhof B, Poulaki V (2001) Inhibition of inflammatory corneal angiogenesis by TNP-470. Invest Ophthalmol Vis Sci 42:2510–2516PubMedGoogle Scholar
  2. 2.
    Chang JH, Gabison EE, Kato T, Azar DT (2001) Corneal neovascularization. Curr Opin Ophthalmol 12:242–249CrossRefPubMedGoogle Scholar
  3. 3.
    Cursiefen C, Schlotzer-Schrehardt U, Kuchle M et al (2002) Lymphatic vessels in vascularized human corneas: immunohistochemical investigation using LYVE-1 and podoplanin. Invest Ophthalmol Vis Sci 43:2127–2135PubMedGoogle Scholar
  4. 4.
    Cursiefen C, Chen L, Dana MR, Streilein JW (2003) Corneal lymphangiogenesis: evidence, mechanisms, and implications for corneal transplant immunology. Cornea 22:273–281CrossRefPubMedGoogle Scholar
  5. 5.
    Regina M, Zimmerman R, Malik G, Gausas R (2007) Lymphangiogenesis concurrent with haemangiogenesis in the human cornea. Clin Exp Ophthalmol 35:541–544CrossRefPubMedGoogle Scholar
  6. 6.
    Haynes WL, Proia AD, Klintworth GK (1989) Effect of inhibitors of arachidonic acid metabolism on corneal neovascularization in the rat. Invest Ophthalmol Vis Sci 30:1588–1593PubMedGoogle Scholar
  7. 7.
    Lipman RM, Epstein RJ, Hendricks RL (1992) Suppression of corneal neovascularization with cyclosporine. Arch Ophthalmol 110:405–407CrossRefPubMedGoogle Scholar
  8. 8.
    Joussen AM, Kruse FE, Volcker HE, Kirchhof B (1999) Topical application of methotrexate for inhibition of corneal angiogenesis. Graefes Arch Clin Exp Ophthalmol 237:920–927CrossRefPubMedGoogle Scholar
  9. 9.
    Shi W, Gao H, Xie L, Wang S (2006) Sustained intraocular rapamycin delivery effectively prevents high-risk corneal allograft rejection and neovascularization in rabbits. Invest Ophthalmol Vis Sci 47:3339–3344CrossRefPubMedGoogle Scholar
  10. 10.
    Lepri A, Benelli U, Bernardini N et al (1994) Effect of low molecular weight heparan sulphate on angiogenesis in the rat cornea after chemical cauterization. J Ocul Pharmacol 10:273–280CrossRefPubMedGoogle Scholar
  11. 11.
    Xiao O, Xie ZL, Lin BW, Yin X, Pi RB, Zhou SY (2012) Minocycline inhibits alkali burn-induced corneal neovascularization in mice. PLoS ONE 7:1–9Google Scholar
  12. 12.
    Dan L, Shi-long Y, Miao-li L et al (2008) Inhibitory effect of oral doxycycline on neovascularization in a rat corneal alkali burn model of angiogenesis. Curr Eye Res 33:653–660CrossRefPubMedGoogle Scholar
  13. 13.
    Su W, Li Z, Lin M, Li Y, He Z et al (2011) The effect of doxycycline temperature-sensitive hydrogel on inhibiting the corneal neovascularization induced by BFGF in rats. Graef Arch Clin Exp 249:421–427CrossRefGoogle Scholar
  14. 14.
    Peyman GA, Kazi AA, Riazi-Esfahani M et al (2006) The effect of combinations of flurbiprofen, low molecular weight heparin, and doxycycline on the inhibition of corneal neovascularization. Cornea 25:582–585CrossRefPubMedGoogle Scholar
  15. 15.
    Chen WL, Lin CT, Lin NT et al (2009) Subconjunctival injection of bevacizumab (Avastin) on corneal neovascularization in different rabbit models of corneal angiogenesis. Invest Ophthalmol Vis Sci 50:1659–1665CrossRefPubMedGoogle Scholar
  16. 16.
    McCowen MC, Callender ME, Lawlis JF (1951) Fumagillin (H-3) a new antibiotic with amebicidal properties. Science 113:202–203CrossRefPubMedGoogle Scholar
  17. 17.
    Killough JH, Magill GB, Smith RC (1952) The treatment of amebiasis with fumagillin. Science 115:71–72CrossRefPubMedGoogle Scholar
  18. 18.
    Ketznelson H, Jamieson CA (1952) Control of Nosema disease of honey bees with fumagillin. Science 115:70CrossRefGoogle Scholar
  19. 19.
    Lowder CY, McMahon JT, Meisler DM et al (1996) Microsporidial keratoconjunctivitis caused by Septata intestinalis in a patient with acquired immunodeficiency syndrome. Am J Ophthalmol 121:715–717CrossRefPubMedGoogle Scholar
  20. 20.
    Vemuganti GK, Garg P, Sharma S, Joseph J, Gopinathan U, Singh S (2005) Is microsporidial keratitis an emerging cause of stromal keratitis? A case series study. BMC Ophthalmol 5:19CrossRefPubMedPubMedCentralGoogle Scholar
  21. 21.
    Diesenhouse MC, Wilson LA, Corrent GF, Visvesvara GS, Grossniklaus HE, Bryan RT (1993) Treatment of microsporidial keratoconjunctivitis with topical fumagillin. Am J Ophthalmol 115:293–298CrossRefPubMedGoogle Scholar
  22. 22.
    Cali A, Meisler DM, Lowder CY et al (1991) Corneal microsporidioses: characterization and identification. J Protozool 38:215–217CrossRefGoogle Scholar
  23. 23.
    Ingber D, Fujita T, Kishimoto S (1990) Synthetic analogues of fumagillin that inhibit angiogenesis and suppress tumor growth. Nature 348:555–557CrossRefPubMedGoogle Scholar
  24. 24.
    Mahoney JM, Waterbury LD (1985) Drug effects on the neovascularization response to silver nitrate cauterization of the rat cornea. Curr Eye Res 4:531–535CrossRefPubMedGoogle Scholar
  25. 25.
    Oh SJ, Jeltsch MM, Birkenhager R et al (1997) VEGF and VEGF-C: specific induction of angiogenesis and lymphangiogenesis in the differentiated avian chorioallantoic membrane. Dev Biol 188:96–109CrossRefPubMedGoogle Scholar
  26. 26.
    Skobe M, Hawighorst T, Jackson DG et al (2001) Induction of tumor lymphangiogenesis by VEGF-C promotes breast cancer metastasis. Nat Med 7:192–198CrossRefPubMedGoogle Scholar
  27. 27.
    Ling S, Lin H, Liang L et al (2009) Development of new lymphatic vessels in alkali burned corneas. Acta Ophthalmol 87:315–322CrossRefPubMedGoogle Scholar
  28. 28.
    Lanternier F, Boutboul D, Menotti J et al (2009) Microsporidiosis in solid organ transplant recipients: two Enterocytozoon bieneusi cases and review. Transpl Infect Dis 11:83–88CrossRefPubMedGoogle Scholar
  29. 29.
    Molina JM, Goguel J, Sarfati C et al (2000) Trial of oral fumagillin for the treatment of intestinal microsporidiosis in patients with HIV infection. AIDS 14:1341–1348CrossRefPubMedGoogle Scholar
  30. 30.
    Molina JM, Tourneur M, Sarfati C, Chevret S, de Gouvello A, Gobert JG, Balkan S, Derouin F, Agence Nationale de Recherches sur le SIDA 090 Study Group (2002) Fumagillin treatment of intestinal microsporidiosis. N Engl J Med 346:1963–1969CrossRefPubMedPubMedCentralGoogle Scholar
  31. 31.
    Loh RS, Chan CM, Ti SE, Lim L, Chan KS, Tan DT (2009) Emerging prevalence of microsporidial keratitis epidemiology, clinical features and management. Ophthalmology 116:2348–2353CrossRefPubMedGoogle Scholar
  32. 32.
    Griffith EC, Su Z, Niwayama S, Ramsay CA, Chang YH, Liu JO (1998) Molecular recognition of angiogenesis inhibitors fumagillin and ovalicin by methionine aminopeptidase 2. Proc Natl Acad Sci USA 95:15183–15188CrossRefPubMedGoogle Scholar
  33. 33.
    Zhang P, Nicholson DE, Bujnicki JM et al (2002) Angiogenesis inhibitors specific for methionine aminopeptidase 2 as drugs for malaria and leishmaniasis. J Biomed Sci 9:34–40CrossRefPubMedGoogle Scholar
  34. 34.
    Kieser A, Weich HA, Brandner G, Marme D, Kolch W (1994) Mutant p53 potentiates protein kinase C induction of vascular endothelial growth factor expression. Oncogene 9:963–969PubMedGoogle Scholar
  35. 35.
    Emoto M, Ishiguro M, Iwasaki H, Kikuchi M, Kawarabayashi T (2000) TNP-470 inhibits growth and the production of vascular endothelial growth factor of uterine carcinosarcoma cells in vitro. Anticancer Res 20:601–604PubMedGoogle Scholar
  36. 36.
    Kanno T, Uehara T, Osawa M et al (2015) Fumagillin, a potent angiogenesis inhibitor, induces Kaposi sarcoma-associated herpesvirus replication in primary effusion lymphoma cells. Biochem Biophys Res Commun 463:1267–1272CrossRefPubMedGoogle Scholar
  37. 37.
    Yanase T, Tamura M, Fujita K, Kodama S, Tanaka K (1993) Inhibitory effect of angiogenesis inhibitor TNP-470 on tumor growth and metastasis of human cell lines in vitro and in vivo. Cancer Res 53:2566–2570PubMedGoogle Scholar

Copyright information

© Springer Nature B.V. 2018

Authors and Affiliations

  1. 1.Department of OphthalmologyBakirkoy Dr. Sadi Konuk Training and Research HospitalIstanbulTurkey

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