Manufacturing of Trojan Horse-TNFR Decoy Receptor Fusion Protein

DESCRIPTION (provided by applicant): Tumor necrosis factor (TNF)-alpha plays a pro-inflammatory role in aging-related neurodegenerative diseases, such as Alzheimer's disease or Parkinson's disease. The biologic TNF inhibitors (TNFI), such as the TNF decoyreceptor, cannot be developed for brain diseases, because the TNFIs are large molecules that do not cross the blood-brain barrier (BBB). The present work continues the drug development of a BBB-penetrating biologic TNFI, which is a re-engineered form of the human type II TNF receptor (TNFR), wherein the TNFR is produced as an IgG fusion protein. The IgG part is a genetically engineered monoclonal antibody (MAb) against the human insulin receptor (HIR). The HIRMAb-TNFR fusion protein is named AGT-110. The HIRMAb part of the HIRMAb-TNFR fusion protein acts as a molecular Trojan horse to ferry the fused decoy receptor across the BBB via receptor-mediated transport on the endogenous BBB insulin receptor. In this phase II SBIR project, the methodology for manufacturing of the HIRMAb-TNFR fusion protein at a large scale suitable for production of clinical trial lots of study drug will be developed. The stably transfected host cell will be cultured in a 50L bioreactor, and the fusion protein will be purified with 1liter columns or protein A affinity chromatography, cation exchange chromatography, and anion exchange chromatography. The identity, purity, potency, safety, and impurities of the drug product will be evaluated by gt15 test methods. The pharmacokinetics,immune response, safety pharmacology, and toxicology will be tested for the first time in adult Rhesus monkeys in an initial dose-ranging study. If successful, this research will provide the basis for a Pre-IND Meeting with the FDA, and entry of AGT-110 drug development into the phases of GLP pharmacology and GMP manufacturing. The overall goal of this work is the development of brain penetrating biologic TNFI, so that the pro-inflammatory effects of TNFalpha in neural disease can be suppressed. PUBLIC HEALTH RELEVANCE PUBLIC HEALTH RELEVANCE: Biologic tumor necrosis factor inhibitors (TNFI), such as the tumor necrosis factor (TNFR) decoy receptor, cannot be developed for aging-related brain diseases, because these large molecule drugs do not cross the blood-brain barrier (BBB). The present research re-engineers the TNFR decoy receptor as a BBB-penetrating IgG fusion protein. The availability of this new brain penetrating IgG-TNFR fusion protein will enable future treatment of TNF-dependent chronic neurodegenerative diseases.