A Therapeutic Uricase with Reduced Immunogenicity Risk and Improved Development Properties

Autor: Flaviu Gruia, Jeffrey N. Miner, Frank Bartnik, David M. Wilson, Luba Grinberg, Ryan Bean, Jane K. Osbourn, Herren Wu, Hui Feng, Lutz Jermutus, James C. Geoghegan, Benoy Chacko, Vidyashankara Iyer, Susan Wilson, David A. Owen, Manuel Baca, Simone M Nicholson, Varnika Roy, Mark Berge, Anna Zacco, Steven Coats, Dongmei Zhou, Ellen O'Connor, Chris Ward, Chi Y. Wu, Michaela Wendeler, Andrew C. Nyborg, Clynn Wilker
Rok vydání: 2016
Předmět:
0301 basic medicine
Gout
Urate Oxidase
Physiology
Swine
T-Lymphocytes
lcsh:Medicine
Pharmacology
Biochemistry
Polyethylene Glycols
Substrate Specificity
White Blood Cells
Database and Informatics Methods
chemistry.chemical_compound
Subcutaneous injection
Animal Cells
Immune Physiology
Medicine and Health Sciences
Post-Translational Modification
lcsh:Science
chemistry.chemical_classification
Immune System Proteins
Multidisciplinary
Calorimetry
Differential Scanning
T Cells
Immunogenicity
Urate oxidase
Hematology
Hydrogen-Ion Concentration
Recombinant Proteins
Body Fluids
Actinobacteria
Blood
Cellular Types
Anatomy
Sequence Analysis
Research Article
Half-Life
Bioinformatics
Immune Cells
Inflammatory Diseases
Immunology
Research and Analysis Methods
03 medical and health sciences
Dogs
Rheumatology
Sequence Motif Analysis
In vivo
Escherichia coli
medicine
Animals
Humans
Arthrobacter
Antigens
Blood Cells
Bacteria
030102 biochemistry & molecular biology
lcsh:R
Organisms
Biology and Life Sciences
Proteins
Cell Biology
Pegylation
medicine.disease
Rats
Kinetics
030104 developmental biology
Enzyme
chemistry
PEGylation
Uric acid
lcsh:Q
Sequence Alignment
Papio
Zdroj: PLoS ONE, Vol 11, Iss 12, p e0167935 (2016)
PLoS ONE
ISSN: 1932-6203
DOI: 10.1371/journal.pone.0167935
Popis: Humans and higher primates are unique in that they lack uricase, the enzyme capable of oxidizing uric acid. As a consequence of this enzyme deficiency, humans have high serum uric acid levels. In some people, uric acid levels rise above the solubility limit resulting in crystallization in joints, acute inflammation in response to those crystals causes severe pain; a condition known as gout. Treatment for severe gout includes injection of non-human uricase to reduce serum uric acid levels. Krystexxa® is a hyper-PEGylated pig-baboon chimeric uricase indicated for chronic refractory gout that induces an immunogenic response in 91% of treated patients, including infusion reactions (26%) and anaphylaxis (6.5%). These properties limit its use and effectiveness. An innovative approach has been used to develop a therapeutic uricase with improved properties such as: soluble expression, neutral pH solubility, high E. coli expression level, thermal stability, and excellent activity. More than 200 diverse uricase sequences were aligned to guide protein engineering and reduce putative sequence liabilities. A single uricase lead candidate was identified, which showed low potential for immunogenicity in >200 human donor samples selected to represent diverse HLA haplotypes. Cysteines were engineered into the lead sequence for site specific PEGylation and studies demonstrated >95% PEGylation efficiency. PEGylated uricase retains enzymatic activity in vitro at neutral pH, in human serum and in vivo (rats and canines) and has an extended half-life. In canines, an 85% reduction in serum uric acid levels was observed with a single subcutaneous injection. This PEGylated, non-immunogenic uricase has the potential to provide meaningful benefits to patients with gout.
Databáze: OpenAIRE
Externí odkaz: