Corneal toxicity of cell-penetrating peptides that inhibit Herpes simplex virus entry.
Cell-penetrating peptides (CPPs) inhibit Herpes simplex virus entry at low micromolar concentrations and may be useful either as prophylactic or therapeutic agents for herpetic keratitis. The aim of this study was to assess the in vitro and in vivo toxicity of three CPPs-EB, TAT-C, and HOM (penetratin)-for the cornea. Incubation of primary (HK320) or immortalized (THK320) human keratocytes with the EB peptide (up to 100 microM), bHOMd (up to 200 microM), or TAT-C (up to 400 microM) resulted in no evidence of toxicity using a formazan dye-reduction assay. Similar results were obtained with a human trabecular meshwork cell line (TM-1), primary human foreskin fibroblasts (DP-9), Vero, and HeLa cells with EB and TATC. The bHOMd peptide showed some toxicity in Vero and HeLa cells, with CC50 values of 70 and 93 microM, respectively. The EB peptide did not inhibit macromolecular synthesis in Vero cells at concentrations below 150 microM, although cell proliferation was blocked at concentrations of EB above 50 microM. In vivo toxicity was assessed by applying peptides in Dulbecco's modified Eagle's medium to the cornea 4 times daily for 7 d. At concentrations 1000 times the IC50 values, the EB and bHOM peptides showed no toxicity, whereas TAT-C caused some mild eyelid swelling. Some slight epithelial cell sloughing was seen with the bKLA peptide in vivo. These results suggest that these CPPs-and EB in particular-have a favorable toxicity profile, and that further development is warranted.
Akkarawongsa
Radeekorn
R
Program in Cell and Molecular Biology, Department of Ophthalmology and Visual Sciences, University of Wisconsin School of Medicine and Public Health, Madison, WI, USA.
Cullinan
Amy E
AE
Zinkel
Andrew
A
Clarin
Joshua
J
Brandt
Curtis R
CR
D000818:Animals; D002352:Carrier Proteins / Q000633:toxicity*; D049109:Cell Proliferation / Q000187:drug effects; D057846:Cell-Penetrating Peptides; D002522:Chlorocebus aethiops; D003315:Cornea / Q000187:drug effects* / Q000473:pathology / Q000821:virology; D005260:Female; D051519:Fibroblast Growth Factor 4 / Q000633:toxicity*; D005347:Fibroblasts / Q000187:drug effects / Q000821:virology; D015696:Gene Products, tat / Q000633:toxicity*; D006367:HeLa Cells / Q000187:drug effects / Q000821:virology; D018259:Herpesvirus 1, Human / Q000187:drug effects* / Q000472:pathogenicity; D006801:Humans; D051379:Mice; D008807:Mice, Inbred BALB C; D010446:Peptide Fragments / Q000633:toxicity*; D014129:Trabecular Meshwork / Q000187:drug effects / Q000821:virology; D014709:Vero Cells / Q000187:drug effects / Q000821:virology
D002352:Carrier Proteins; D057846:Cell-Penetrating Peptides; C496445:FGF4 protein, human; D051519:Fibroblast Growth Factor 4; D015696:Gene Products, tat; D010446:Peptide Fragments; C414409:penetratin