117621 20240319081006.0 doi 10.3390/ijms23094502 sideral 129167 ART-2022-129167 eng Conde-Giménez, M. (orcid)0000-0003-4358-9110 Alchemical Design of Pharmacological Chaperones with Higher Affinity for Phenylalanine Hydroxylase 2022 Access copy available to the general public Unrestricted Phenylketonuria (PKU) is a rare metabolic disease caused by variations in a human gene, PAH, encoding phenylalanine hydroxylase (PAH), and the enzyme converting the essential amino acid phenylalanine into tyrosine. Many PKU-causing variations compromise the conformational stability of the encoded enzyme, decreasing or abolishing its catalytic activity, and leading to an elevated concentration of phenylalanine in the blood, which is neurotoxic. Several therapeutic approaches have been developed to treat the more severe manifestations of the disorder, but they are either not entirely effective or difficult to adhere to throughout life. In a search for novel pharmacological chaperones to treat PKU, a lead compound was discovered (compound IV) that exhibited promising in vitro and in vivo chaperoning activity on PAH. The structure of the PAH-IV complex has been reported. Here, using alchemical free energy calculations (AFEC) on the structure of the PAH-IV complex, we design a new generation of compound IV-analogues with a higher affinity for the enzyme. Seventeen novel analogues were synthesized, and thermal shift and isothermal titration calorimetry (ITC) assays were performed to experimentally evaluate their stabilizing effect and their affinity for the enzyme. Most of the new derivatives bind to PAH tighter than lead compound IV and induce a greater thermostabilization of the enzyme upon binding. Importantly, the correspondence between the calculated alchemical binding free energies and the experimentally determined ¿¿Gb values is excellent, which supports the use of AFEC to design pharmacological chaperones to treat PKU using the X-ray structure of their complexes with the target PAH enzyme. © 2022 by the authors. Licensee MDPI, Basel, Switzerland. info:eu-repo/grantAgreement/ES/DGA/E45-20R info:eu-repo/grantAgreement/ES/DGA/LMP30-18 info:eu-repo/grantAgreement/ES/MINECO/BFU2016-78232-P info:eu-repo/grantAgreement/ES/MINECO/PID2019- 107293GB-I00 info:eu-repo/semantics/openAccess by http://creativecommons.org/licenses/by/3.0/es/ 5.6 2022 BIOCHEMISTRY & MOLECULAR BIOLOGY 66 / 285 = 0.232 2022 Q1 T1 CHEMISTRY, MULTIDISCIPLINARY 52 / 178 = 0.292 2022 Q2 T1 1.154 2022 Medicine (miscellaneous) 2022 Q1 Physical and Theoretical Chemistry 2022 Q1 Computer Science Applications 2022 Q1 Inorganic Chemistry 2022 Q1 Spectroscopy 2022 Q1 Organic Chemistry 2022 Q1 Molecular Biology 2022 Q2 Catalysis 2022 Q2 7.8 2022 info:eu-repo/semantics/article info:eu-repo/semantics/publishedVersion Galano-Frutos, J. J. Universidad de Zaragoza (orcid)0000-0002-1896-7805 Galiana-Cameo, M. Universidad de Zaragoza (orcid)0000-0002-2043-4864 Mahía Moros, A. (orcid)0000-0002-8839-6796 Victor, B. L. Salillas Berges, S. Universidad de Zaragoza (orcid)0000-0003-0195-5434 Velázquez-Campoy, A. Universidad de Zaragoza (orcid)0000-0001-5702-4538 Brito, R. M. M. Gálvez Lafuente, J. A. Universidad de Zaragoza (orcid)0000-0002-4972-7476 Díaz-de-villegas, M. D. (orcid)0000-0001-9033-8459 Sancho Sanz, J. Universidad de Zaragoza (orcid)0000-0002-2879-9200 2010 760 Universidad de Zaragoza Dpto. Química Inorgánica Área Química Inorgánica 1011 630 Universidad de Zaragoza Dpto. Microb.Ped.Radio.Sal.Pú. Área Microbiología 2013 765 Universidad de Zaragoza Dpto. Química Orgánica Área Química Orgánica 1002 060 Universidad de Zaragoza Dpto. Bioq.Biolog.Mol. Celular Área Bioquímica y Biolog.Mole. 23, 9 (2022), 4502 [21 pp] Int. j. mol. sci. International Journal of Molecular Sciences 1661-6596 8421771 http://zaguan.unizar.es/record/117621/files/texto_completo.pdf Versión publicada 2806718 http://zaguan.unizar.es/record/117621/files/texto_completo.jpg?subformat=icon icon Versión publicada oai:zaguan.unizar.es:117621 articulos driver 2024-03-18-14:36:47 ARTICLE