Pharmaceutical Research

, 35:173 | Cite as

Safety Assessment of Formulation Vehicles Following Intravitreal Administration in Rabbits

  • Shirley A. AguirreEmail author
  • Hovhannes J. Gukasyan
  • Husam S. Younis
  • Wenhu Huang
Research Paper
Part of the following topical collections:
  1. Ophthalmic Drug Discovery and Development



Evaluate 21 formulation vehicles administered to rabbits after intravitreal injection for tolerability and safety.


Forty-two Dutch Belted rabbits were anesthetized, and the eyes received a single intravitreal injection of the excipient formulation. Clinical signs and ocular irritation responses were recorded twice daily for 7 days and microscopic evaluation of the eyes, optic nerve, and eyelids was completed at 1-week post treatment.


Saline (≥ 300 mOsm and ≤ 592 mOsm at pH 7.0 or 300 mOsm at pH 8.0) and 10 formulation excipients; (10% w/v PEG 3350 at pH 7.4, 1% polysorbate 21 at pH 7.4, PVA at pH 7.0, 0.2% polysorbate 80 at pH 7.2, 0.2% Pluronic F108® at pH 7.3, 2%, 100 mM sodium sulfate at pH 3.2, 2 mM sodium glycocholate at pH 7.4, and 275 mM D-mannitol pH 7.0 in sterile water, and 100 mM sodium phosphate in combination with 0.9% NaCl 300 mOsm and 0.01% or 0.05% polysorbate 80 at pH 7.4) considered as formulation vehicles for intravitreal injectables, were well-tolerated in rabbits. Clinical signs were transient and microscopic changes were not observed.


Of the 21 formulation vehicles evaluated, 10 formulation vehicles were well-tolerated in rabbits and feasible candidates for future investigations.

Key words

excipients formulations intravitreal rabbit safety 



Blood retinal barrier


Sodium glycocholate

Captisol® or SBE-β-CD





Ethyl cellulose


Ganglion cell layer


Inner nuclear layer


Inner plexiform layer


Inner segment of photoreceptor layer



Medisorb® or PLGA

Polylactic-co-glycolic acid


Myelinated nerve fiber




Sodium sulfate


Sodium phosphate


Sodium citrate


Sodium chloride


Outer nuclear layer


Outer plexiform layer


Outer segment of photoreceptor layer


Polyethylene glycol

Pluronic® F108 or Poloxamer 338

Polyethylene-polypropylene glycol


Polyvinyl alcohol


Retinal pigment epithelium

Solutol® HS 15



Acknowledgments and Disclosures

The authors would like to thank Winston Evering for review of the manuscript and helpful discussions, Carlos Esparza, Jessica Frey and Anthony Wong for histology assistance, Walter Collette III for in vivo support and Cristiana Janssen for graphic arts assistance.


  1. 1.
    Hughes PM, Olejnik O, Chang-Lin JE, Wilson CG. Topical and systemic drug delivery to the posterior segments. Adv Drug Deliv Rev. 2005;57(14):2010–32.CrossRefPubMedGoogle Scholar
  2. 2.
    Rowe-Rendleman CL, Durazo SA, Kompella UB, Rittenhouse KD, Di Polo A, Weiner AL, et al. Drug and gene delivery to the back of the eye: from bench to bedside. Invest Ophthalmol Vis Sci. 2014;55(4):2714–30.CrossRefPubMedPubMedCentralGoogle Scholar
  3. 3.
    Jager RD, Aiello LP, Patel SC, Cunningham ET Jr. Risks of intravitreous injection: a comprehensive review. Retina. 2004;24(5):676–98.CrossRefPubMedGoogle Scholar
  4. 4.
    Gopinathan S, O’Neill E, Rodriguez LA, Champ R, Phillips M, Nouraldeen A, et al. In vivo toxicology of excipients commonly employed in drug discovery in rats. J Pharmacol Toxicol Methods. 2013;68(2):284–95.CrossRefPubMedGoogle Scholar
  5. 5.
    Freeman DJ, Niven RW. The influence of sodium glycocholate and other additives on the in vivo transfection of plasmid DNA in the lungs. Pharm Res. 1996;13(2):202–9.CrossRefPubMedGoogle Scholar
  6. 6.
    Wilson AG, Nouraldeen A, Gopinathan S. A new paradigm for improving oral absorption of drugs in discovery: role of physicochemical properties, different excipients and the pharmaceutical scientist. Future Med Chem. 2010;2(1):1–5.CrossRefPubMedGoogle Scholar
  7. 7.
    Pifferi G, Restani P. The safety of pharmaceutical excipients. Farmaco. 2003;58(8):541–50.CrossRefPubMedGoogle Scholar
  8. 8.
    Qaddoumi MG, Gukasyan HJ, Davda J, Labhasetwar V, Kim KJ, Lee VHL. Clathrin and caveolin-1 expression in primary pigmented rabbit conjunctival epithelial cells: Role of PLGA nanoparticle endocytosis. Mol Vis. 2003;9:559–68.PubMedGoogle Scholar
  9. 9.
    Chen JJ, Ebmeiser SE, Sutherland WM, Ghazi NG. Potential penetration of topical ranibizumab (Lucentis) in the rabbit eye. Eye (Lond). 2011;25(11):1504–11.CrossRefGoogle Scholar
  10. 10. OECD Guideline For The Testing Of Chemicals. Proposal for an unpdated TG 405. Acute Eye Irritiation/Corrosion. OECD; 1987. p. 1–15.Google Scholar
  11. 11.
    Aguirre SA, Collette W 3rd, Gukasyan HJ, Huang W. An assessment of the ocular safety of excipient maleic acid following intravitreal injection in rabbits. Toxicol Pathol. 2012;40(5):797–806.CrossRefPubMedGoogle Scholar
  12. 12.
    Lorget F, Parenteau A, Carrier M, Lambert D, Gueorguieva A, Schuetz C, et al. Characterization of the pH and Temperature in the Rabbit, Pig, and Monkey Eye: Key Parameters for the Development of Long-Acting Delivery Ocular Strategies. Mol Pharm. 2015;23:23.Google Scholar
  13. 13.
    de Oliveira Dias JR, Xavier CO, Maia A, de Moraes NS, Meyer C, Farah ME, et al. Intravitreal injection of ziv-aflibercept in patient with refractory age-related macular degeneration. Ophthalmic Surg Lasers Imaging Retina. 2015;46(1):91–4.CrossRefPubMedGoogle Scholar
  14. 14.
    Marmor MF, Martin LJ, Tharpe S. Osmotically induced retinal detachment in the rabbit and primate. Electron miscoscopy of the pigment epithelium. Invest Ophthalmol Vis Sci. 1980;19(9):1016–29.PubMedGoogle Scholar
  15. 15.
    Hosoya K, Makihara A, Tsujikawa Y, Yoneyama D, Mori S, Terasaki T, et al. Roles of inner blood-retinal barrier organic anion transporter 3 in the vitreous/retina-to-blood efflux transport of p-aminohippuric acid, benzylpenicillin, and 6-mercaptopurine. J Pharmacol Exp Ther. 2009;329(1):87–93.CrossRefPubMedGoogle Scholar
  16. 16.
    Yanoff M, Sassini JW. Ocular Pathology. In: Gabbedy R, editor. Ocular pathology. 6th ed: Mosby Elesevier; 2009. p. 485.Google Scholar
  17. 17.
    Webster R, Didier E, Harris P, Siegel N, Stadler J, Tilbury L, et al. PEGylated proteins: evaluation of their safety in the absence of definitive metabolism studies. Drug Metab Dispos. 2007;35(1):9–16.CrossRefPubMedGoogle Scholar
  18. 18.
    Dow. Technical Data Sheet CarboxwaxTM Polyethylene Glycol (PEG) 400. In: Dow, editor. 2011.Google Scholar
  19. 19.
    Patel S, Barnett JM, Kim SJ. Retinal Toxicity of Intravitreal Polyethylene Glycol 400. J Ocul Pharmacol Ther. 2015;32(2):97–101.CrossRefPubMedGoogle Scholar
  20. 20.
    Lyzogubov VV, Tytarenko RG, Liu J, Bora NS, Bora PS. Polyethylene glycol (PEG)-induced mouse model of choroidal neovascularization. J Biol Chem. 2011;286(18):16229–37.CrossRefPubMedPubMedCentralGoogle Scholar
  21. 21.
    Hayreh SS. Posterior drainage of the intraocular fluid from the vitreous. Exp Eye Res. 1966;5:123–44.CrossRefGoogle Scholar

Copyright information

© Springer Science+Business Media, LLC, part of Springer Nature 2018

Authors and Affiliations

  1. 1.Pfizer Inc., Drug Safety Research and DevelopmentLa Jolla LaboratoriesSan DiegoUSA
  2. 2.Virchow Toxpath LLCPrescottUSA
  3. 3.AllerganIrvineUSA
  4. 4.NGM Biopharmaceuticals, Inc.South San FranciscoUSA

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