title: Poly(ethylene glycol) (PEG) conjugated arginine deiminase: effects of PEG formulations on its pharmacological properties.
abstract: Some tumors, such as melanomas and hepatocellular carcinomas, have a unique nutritional requirement for arginine. Thus, enzymatic degradation of extracellular arginine is one possible means for inhibiting these tumors. Arginine deiminase is an arginine degrading enzyme (ADI) that has been studied as an anti-cancer enzyme. However, ADI has a short serum half-life and, as a microbial enzyme, is highly immunogenic. Formulation of other therapeutic proteins with poly(ethylene glycol) (PEG) has overcome these problems. Here, ADI-PEGs were synthesized using PEGs of varying size, structure (linear or branched chain) and linker chemistries. All ADI-PEGs retained approximately 50% of enzyme activity when PEG was covalently attached to approximately 40% of the primary amines irrespective of the PEG molecular weight or attachment chemistry used. However, it was observed that, as the PEG size increases to 20 kDa, there was a corresponding increase in the pharmacokinetic (pK) and pharmacodynamic (pD) properties of the formulation. Variation in PEG linker or structure, or the use of PEGs >20,000 mw, did not affect the pK or pD. As has been shown with other therapeutic proteins, repeated injection of ADI-PEG into experimental animals resulted in significantly lower titers of antibodies against this protein than unmodified ADI. These data suggest that formulation of ADI with PEG of 20,000 mw results is the optimal method for formulating this promising therapeutic agent.
authors.affiliation: Department of Biology, T.H. Morgan Building, University of Kentucky, Lexington, KY 40506, USA.
authors.forename: Frederick W
authors.initials: FW
authors.lastname: Holtsberg
authors.forename: Charles Mark
authors.initials: CM
authors.lastname: Ensor
authors.forename: Marion R
authors.initials: MR
authors.lastname: Steiner
authors.forename: John S
authors.initials: JS
authors.lastname: Bomalaski
authors.forename: Mike A
authors.initials: MA
authors.lastname: Clark
mesh_terms: D000818:Animals; D000941:Antigens / Q000008:administration & dosage / Q000276:immunology; D019540:Area Under Curve; D001120:Arginine / Q000097:blood; D002626:Chemistry, Pharmaceutical; D004926:Escherichia coli / Q000201:enzymology; D005260:Female; D006207:Half-Life; D006867:Hydrolases / Q000737:chemistry / Q000276:immunology / Q000493:pharmacokinetics* / Q000494:pharmacology; D051379:Mice; D010316:Particle Size; D011092:Polyethylene Glycols / Q000008:administration & dosage / Q000737:chemistry / Q000493:pharmacokinetics* / Q000494:pharmacology; D011817:Rabbits
chemical_list: D000941:Antigens; D011092:Polyethylene Glycols; D001120:Arginine; D006867:Hydrolases; C019723:arginine deiminase