PUBLICATION
A Trivalent Enzymatic System for Uricolytic Therapy of HPRT Deficiency and Lesch-Nyhan Disease
- Authors
- Ronda, L., Marchetti, M., Piano, R., Liuzzi, A., Corsini, R., Percudani, R., Bettati, S.
- ID
- ZDB-PUB-170517-6
- Date
- 2017
- Source
- Pharmaceutical research 34(7): 1477-1490 (Journal)
- Registered Authors
- Keywords
- Lesch-Nyhan disease, enzyme therapy, gout, hyperuricemia, polyethylene glycol, purine degradation
- MeSH Terms
-
- Allantoin/chemistry
- Amidohydrolases/chemistry*
- Animals
- Carboxy-Lyases/chemistry*
- Enzyme Therapy
- Humans
- Hyperuricemia/drug therapy
- Hypoxanthine Phosphoribosyltransferase/deficiency*
- Lesch-Nyhan Syndrome/drug therapy*
- Molecular Weight
- Polyethylene Glycols/chemistry*
- Recombinant Proteins/chemistry
- Solubility
- Stereoisomerism
- Urate Oxidase/chemistry*
- Uric Acid/chemistry
- Uricosuric Agents/chemistry*
- Zebrafish
- PubMed
- 28508122 Full text @ Pharm. Res.
Citation
Ronda, L., Marchetti, M., Piano, R., Liuzzi, A., Corsini, R., Percudani, R., Bettati, S. (2017) A Trivalent Enzymatic System for Uricolytic Therapy of HPRT Deficiency and Lesch-Nyhan Disease. Pharmaceutical research. 34(7):1477-1490.
Abstract
Purpose Because of the evolutionary loss of the uricolytic pathway, humans accumulate poorly soluble urate as the final product of purine catabolism. Restoration of uricolysis through enzyme therapy is a promising treatment for severe hyperuricemia caused by deficiency of hypoxanthine-guanine phosphoribosyltransferase (HPRT). To this end, we studied the effect of PEG conjugation on the activity and stability of the enzymatic complement required for conversion of urate into the more soluble (S)-allantoin.
Methods We produced in recombinant form three zebrafish enzymes required in the uricolytic pathway. We carried out a systematic study of the effect of PEGylation on the function and stability of the three enzymes by varying PEG length, chemistry and degree of conjugation. We assayed in vitro the uricolytic activity of the PEGylated enzymatic triad.
Results We defined conditions that allow PEGylated enzymes to retain native-like enzymatic activity even after lyophilization or prolonged storage. A combination of the three enzymes in an appropriate ratio allowed efficient conversion of urate to (S)-allantoin with no accumulation of intermediate metabolites.
Conclusions Pharmaceutical restoration of the uricolytic pathway is a viable approach for the treatment of severe hyperuricemia.
Genes / Markers
Expression
Phenotype
Mutations / Transgenics
Human Disease / Model
Sequence Targeting Reagents
Fish
Orthology
Engineered Foreign Genes
Mapping