Novel pharmacological chaperones that correct phenylketonuria in mice

TY - JOUR

T1 - Novel pharmacological chaperones that correct phenylketonuria in mice

AU - Santos-Sierra, Sandra

AU - Kirchmair, Johannes

AU - Perna, Anna M

AU - Reiss, Dunja

AU - Kemter, Kristina

AU - Röschinger, Wulf

AU - Glossmann, Hartmut

AU - Gersting, Søren W

AU - Muntau, Ania C

AU - Wolber, Gerhard

AU - Lagler, Florian B

PY - 2012/4/15

Y1 - 2012/4/15

N2 - Phenylketonuria (PKU) is caused by inherited phenylalanine-hydroxylase (PAH) deficiency and, in many genotypes, it is associated with protein misfolding. The natural cofactor of PAH, tetrahydrobiopterin (BH(4)), can act as a pharmacological chaperone (PC) that rescues enzyme function. However, BH(4) shows limited efficacy in some PKU genotypes and its chemical synthesis is very costly. Taking an integrated drug discovery approach which has not been applied to this target before, we identified alternative PCs for the treatment of PKU. Shape-focused virtual screening of the National Cancer Institute's chemical library identified 84 candidate molecules with potential to bind to the active site of PAH. An in vitro evaluation of these yielded six compounds that restored the enzymatic activity of the unstable PAHV106A variant and increased its stability in cell-based assays against proteolytic degradation. During a 3-day treatment study, two compounds (benzylhydantoin and 6-amino-5-(benzylamino)-uracil) substantially improved the in vivo Phe oxidation and blood Phe concentrations of PKU mice (Pah(enu1)). Notably, benzylhydantoin was twice as effective as tetrahydrobiopterin. In conclusion, we identified two PCs with high in vivo efficacy that may be further developed into a more effective drug treatment of PKU.

AB - Phenylketonuria (PKU) is caused by inherited phenylalanine-hydroxylase (PAH) deficiency and, in many genotypes, it is associated with protein misfolding. The natural cofactor of PAH, tetrahydrobiopterin (BH(4)), can act as a pharmacological chaperone (PC) that rescues enzyme function. However, BH(4) shows limited efficacy in some PKU genotypes and its chemical synthesis is very costly. Taking an integrated drug discovery approach which has not been applied to this target before, we identified alternative PCs for the treatment of PKU. Shape-focused virtual screening of the National Cancer Institute's chemical library identified 84 candidate molecules with potential to bind to the active site of PAH. An in vitro evaluation of these yielded six compounds that restored the enzymatic activity of the unstable PAHV106A variant and increased its stability in cell-based assays against proteolytic degradation. During a 3-day treatment study, two compounds (benzylhydantoin and 6-amino-5-(benzylamino)-uracil) substantially improved the in vivo Phe oxidation and blood Phe concentrations of PKU mice (Pah(enu1)). Notably, benzylhydantoin was twice as effective as tetrahydrobiopterin. In conclusion, we identified two PCs with high in vivo efficacy that may be further developed into a more effective drug treatment of PKU.

KW - Animals

KW - Binding Sites

KW - Biopterin

KW - Catalytic Domain

KW - Cell Line, Tumor

KW - Cell Survival

KW - Drug Discovery

KW - Drug Evaluation, Preclinical

KW - Enzyme Stability

KW - Humans

KW - Hydantoins

KW - Mice

KW - Oxidation-Reduction

KW - Phenylalanine

KW - Phenylalanine Hydroxylase

KW - Phenylketonurias

KW - Protein Folding

KW - Small Molecule Libraries

KW - Uracil

U2 - 10.1093/hmg/dds001

DO - 10.1093/hmg/dds001

M3 - SCORING: Journal article

C2 - 22246293

VL - 21

SP - 1877

EP - 1887

JO - HUM MOL GENET

JF - HUM MOL GENET

SN - 0964-6906

IS - 8

ER -