title: Pharmacological chaperones as a potential therapeutic option in methylmalonic aciduria cblB type.
abstract: Methylmalonic aciduria (MMA) cblB type is caused by mutations in the MMAB gene. This encodes the enzyme ATP:cob(I)alamin adenosyltransferase (ATR), which converts reduced cob(I)alamin to an active adenosylcobalamin cofactor. We recently reported the presence of destabilizing pathogenic mutations that retain some residual ATR activity. The aim of the present study was to seek pharmacological chaperones as a tailored therapy for stabilizing the ATR protein. High-throughput ligand screening of over 2000 compounds was performed; six were found to enhance the thermal stability of purified recombinant ATR. Further studies using a well-established bacterial system in which the recombinant ATR protein was expressed in the presence of these six compounds, showed them all to increase the stability of the wild-type ATR and the p.Ile96Thr mutant proteins. Compound V (N-{[(4-chlorophenyl)carbamothioyl]amino}-2-phenylacetamide) significantly increased this stability and did not act as an inhibitor of the purified protein. Importantly, compound V increased the activity of ATR in patient-derived fibroblasts harboring the destabilizing p.Ile96Thr mutation in a hemizygous state to within control range. When cobalamin was coadministrated with compound V, mutant ATR activity further improved. Oral administration of low doses of compound V to C57BL/6J mice for 12 days, led to increase in steady-state levels of ATR protein in liver and brain (disease-relevant organs). These results hold promise for the clinical use of pharmacological chaperones in MMA cblB type patients harboring chaperone-responsive mutations.
authors.affiliation: Centro de Diagnóstico de Enfermedades Moleculares, Centro de Biología Molecular-SO, UAM-CSIC, Universidad Autónoma de Madrid, Campus de Cantoblanco, Madrid, Spain.
authors.forename: Ana
authors.initials: A
authors.lastname: Jorge-Finnigan
authors.forename: Sandra
authors.initials: S
authors.lastname: Brasil
authors.forename: Jarl
authors.initials: J
authors.lastname: Underhaug
authors.forename: Pedro
authors.initials: P
authors.lastname: Ruíz-Sala
authors.forename: Begoña
authors.initials: B
authors.lastname: Merinero
authors.forename: Ruma
authors.initials: R
authors.lastname: Banerjee
authors.forename: Lourdes R
authors.initials: LR
authors.lastname: Desviat
authors.forename: Magdalena
authors.initials: M
authors.lastname: Ugarte
authors.forename: Aurora
authors.initials: A
authors.lastname: Martinez
authors.forename: Belén
authors.initials: B
authors.lastname: Pérez
mesh_terms: D019883:Alkyl and Aryl Transferases / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism; D000592:Amino Acid Metabolism, Inborn Errors / Q000188:drug therapy* / Q000378:metabolism; D000818:Animals; D044482:Benzeneacetamides / Q000008:administration & dosage / Q000737:chemistry* / Q000494:pharmacology*; D001665:Binding Sites; D001921:Brain / Q000187:drug effects / Q000201:enzymology; D004795:Enzyme Stability; D005260:Female; D057166:High-Throughput Screening Assays; D006801:Humans; D008099:Liver / Q000187:drug effects / Q000201:enzymology; D051379:Mice; D008810:Mice, Inbred C57BL; D062105:Molecular Docking Simulation; D050505:Mutant Proteins / Q000235:genetics / Q000378:metabolism; D013890:Thiourea / Q000008:administration & dosage / Q000031:analogs & derivatives* / Q000737:chemistry / Q000494:pharmacology; D014805:Vitamin B 12 / Q000008:administration & dosage / Q000494:pharmacology
chemical_list: D044482:Benzeneacetamides; D050505:Mutant Proteins; C587039:N-(((4-chlorophenyl)carbamothioyl)amino)-2-phenylacetamide; D019883:Alkyl and Aryl Transferases; C019920:cob(I)alamin adenosyltransferase; D013890:Thiourea; D014805:Vitamin B 12