Efficacy of a phosphorodiamidate morpholino oligomer antisense compound in the inhibition of corneal transplant rejection in a rat cornea transplant model

J Ocul Pharmacol Ther. 2012 Apr;28(2):194-201. doi: 10.1089/jop.2011.0135. Epub 2011 Dec 7.

Abstract

Purpose: The cornea is one of the most commonly transplanted tissues. The morpholino-oligomer antisense compound AVI-5126 suppresses expression of proto-oncogene c-myc, a key factor in transplant rejection. AVI-5126 was evaluated in a rat cornea transplant model.

Methods: Donor corneas obtained from August x Copenhagen Irish rats were stored in Optisol™ containing 1.0 or 0.5 mg/mL AVI-5126 or Optisol alone for 24 h before transplant. Recipient Lewis rats were treated topically 3x/d with TobraDex™ and with 1.0 or 0.5 mg/mL of AVI-5126 or saline with daily monitoring for rejection using a modified McDonald-Shadduck Slit Lamp Scale. Using the high-performance liquid chromatography technique, the stability of AVI-5126 (0.5 mg/mL) in Optisol was evaluated for 30 days. AVI-5126 corneal transport was measured using Ussing chamber mounted rabbit corneas. The potential ocular toxicity of AVI-5126 (0.5 mg/mL) was evaluated after 8 days of 3x/d topical application in rats and in-vitro by incubation of human corneas for 8 days.

Results: Cornea storage in Optisol containing 1.0 mg/mL AVI-5126 plus post-transplant topical tid AVI-5126 (1.0 mg/mL) application significantly increased graft survival (7.0±1.6 days) versus 5.0±0.8 days for Optisol alone storage plus post-transplant topical tid saline application (P<0.001). After 30 days of storage, no significant degradation of AVI-5126 in Optisol was noted by high-performance liquid chromatography analysis. After 24 h, 5 μg/mL (1% of total dose) crossed the corneas mounted in Ussing chambers. Neither extended topical application of AVI-5126 to rats nor incubation of human corneas in AVI-5126 decreased endothelial cell density.

Conclusions: Graft rejection was significantly delayed after pretransplantation storage of graft corneas in Optisol containing AVI-5126 followed by topical application of AVI-5126 post-transplantation. AVI-5126 was well tolerated, stable, and effectively penetrated the cornea.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Administration, Topical
  • Animals
  • Chromatography, High Pressure Liquid
  • Cornea / drug effects
  • Corneal Transplantation*
  • Dose-Response Relationship, Drug
  • Drug Stability
  • Female
  • Graft Rejection / prevention & control*
  • Graft Survival / drug effects
  • Humans
  • In Vitro Techniques
  • Morpholinos / administration & dosage
  • Morpholinos / adverse effects
  • Morpholinos / therapeutic use*
  • Oligonucleotides, Antisense / administration & dosage
  • Oligonucleotides, Antisense / adverse effects
  • Oligonucleotides, Antisense / therapeutic use*
  • Proto-Oncogene Mas
  • Rabbits
  • Rats
  • Rats, Inbred Lew
  • Treatment Outcome

Substances

  • AVI-5126
  • MAS1 protein, human
  • Morpholinos
  • Oligonucleotides, Antisense
  • Proto-Oncogene Mas