ABSTRACT
The objective of this manuscript is to provide the reader with two examples on how to present an immunogenicity risk assessment for a PEGylated therapeutic as part of Investigational New Drug (IND) application or during other stages of the drug development process. In order to provide context to the bioanalytical strategies used to support the PEGylated therapeutics presented here, a brief summary of information available for marketed PEGylated biologics is provided. Two case studies are presented, a PEGylated enzyme and a PEGylated growth factor. For the former, the risk assessment covers how to deal with a narrow therapeutic window and suggestions to utilize a PD marker as surrogate for neutralizing antibody assessments in Phase I. The latter has recommendations on additional analytes that should be monitored to mitigate risk of immunogenicity to endogenous counterparts.
Subject(s)
Antibodies, Neutralizing/immunology , Biological Products/immunology , Hepatocyte Growth Factor/immunology , Phenylalanine Ammonia-Lyase/immunology , Polyethylene Glycols , Succinimides/immunology , Animals , Biological Products/chemistry , Biological Products/toxicity , Drug Compounding , Hepatocyte Growth Factor/chemistry , Hepatocyte Growth Factor/toxicity , Humans , Phenylalanine Ammonia-Lyase/chemistry , Phenylalanine Ammonia-Lyase/toxicity , Polyethylene Glycols/chemistry , Polyethylene Glycols/toxicity , Risk Assessment , Succinimides/chemistry , Succinimides/toxicityABSTRACT
PURPOSE: To investigate the antiangiogenic effect of topical application of H-KI20, a novel 20-amino acid peptide from the hepatocyte growth factor, on 2 animal models of corneal neovascularization (NV), and its possible toxic effects on the cornea and conjunctiva. METHODS: The antiangiogenic effect of topical H-KI20 in vivo was studied on corneal NV induced by a mouse corneal micropocket assay and rat intrastromal suture model. In each model, H-KI20, scrambled control peptide H-KI20S, bevacizumab, and phosphate buffer solution (PBS) were applied topically 4 times a day. Corneal NV was examined, photographed, and analyzed. Histological analysis of the corneas was performed. Tear film breakup time and gross and histological examinations were used to study the possible toxicity of topical H-KI20. RESULTS: Topical application of H-KI20 significantly inhibited corneal NV induced by vascular endothelial growth factor (VEGF), and intrastromal suture (P < 0.01 vs. the PBS group), and the area of corneal NV was suppressed by 80.3% and 83.6%, respectively (PBS group as 100%). No significant difference was found between 1.0 mg/mL H-KI20 and 10 mg/mL bevacizumab (P > 0.05). Both hematoxylin and eosin and CD34 staining revealed fewer new blood vessels in the H-KI20 and bevacizumab groups. Tear film breakup time and histological examinations showed that H-KI20 had no obvious toxic effects on the cornea and conjunctiva in vivo. CONCLUSIONS: The novel peptide H-KI20 is an effective and safe inhibitor of corneal NV. It may provide a promising alternative for ocular topical antiangiogenic therapy.
Subject(s)
Angiogenesis Inhibitors/therapeutic use , Corneal Neovascularization/prevention & control , Disease Models, Animal , Hepatocyte Growth Factor/therapeutic use , Peptide Fragments/therapeutic use , Administration, Topical , Amino Acid Sequence , Angiogenesis Inhibitors/chemical synthesis , Angiogenesis Inhibitors/toxicity , Animals , Bevacizumab/therapeutic use , Conjunctiva/drug effects , Cornea/drug effects , Corneal Neovascularization/pathology , Hepatocyte Growth Factor/chemical synthesis , Hepatocyte Growth Factor/chemistry , Hepatocyte Growth Factor/toxicity , Male , Mice , Mice, Inbred C57BL , Molecular Sequence Data , Ophthalmic Solutions , Peptide Fragments/chemical synthesis , Peptide Fragments/toxicity , Peptides/chemical synthesis , Peptides/therapeutic use , Peptides/toxicity , Rats , Rats, Sprague-Dawley , Vascular Endothelial Growth Factor A/antagonists & inhibitorsABSTRACT
Effects of hepatocyte growth factor were investigated in a two-stage rat liver carcinogenesis protocol. Male F344 rats were first treated with diethylnitrosamine (200 mg/kg, i.p.) and then, starting two weeks later, with N-ethyl-N-hydroxyethylnitrosamine (EHEN) for 6 weeks at a dose of 0.01% in drinking water. Hepatocyte growth factor, which was injected i.v. at a dose of 200 micrograms/kg body weight one (at week 3) or two times (at weeks 3 and 4) during EHEN administration, significantly increased the development of preneoplastic glutathione S-transferase placental form-positive foci. Although the observed effects of hepatocyte growth factor were weaker than that of the two-thirds partial hepatectomy (PH) performed at week 3, the present results suggest that the enhancing effects of PH performed during the promotion stage may be largely mediated through induction of hepatocyte growth factor.