Identification of novel 11β-HSD1 inhibitors by combined ligand- and structure-based virtual screening.
11 beta-hydroxysteroid dehydrogenase type 1 (11β-HSD1) converts cortisone to cortisol in a NADPH dependent manner. Overexpression of 11β-HSD1 in key metabolic tissues is related to the development of type 2 diabetes, obesity, hypertension and metabolic syndrome. Using crystal structures of human 11β-HSD1 in complex with inhibitors as source of structural information, a combined ligand and structure-based virtual screening approach was implemented to identify novel 11β-HSD1 inhibitors. A selected group of compounds was identified in silico and further evaluated in cell-based assays for cytotoxicity and 11β-HSD1 mediated cortisol production inhibitory capacity. The expression of 11β-HSD1 and 11β-HSD2 in human LS14 adipocytes was assessed during differentiation. Biological evaluation of 39 compounds in adipocytes and steroids quantification by HPLC-MS/MS identify 4 compounds that exhibit 11β-HSD1 mediated cortisol production inhibitory activity with potencies in the micromolar range. Two compounds showed to be selective for the 11β-HSD1 reductase activity and over 11β-HSD2 isoform, and thus represent novel leads for the development of more active derivatives with higher efficacies targeting intracellular cortisol levels in type 2 diabetes and metabolic syndrome.
Molecular Endocrinology Laboratory, Department of Endocrinology, School of Medicine, Pontificia Universidad Catolica de Chile, Santiago, Chile.
Carlos F
CF
Lagos
Molecular Endocrinology Laboratory, Department of Endocrinology, School of Medicine, Pontificia Universidad Catolica de Chile, Santiago, Chile.
Andrea
A
Vecchiola
Department of Clinical Laboratories, School of Medicine, Pontificia Universidad Catolica de Chile, Santiago, Chile.
Fidel
F
Allende
Molecular Endocrinology Laboratory, Department of Endocrinology, School of Medicine, Pontificia Universidad Catolica de Chile, Santiago, Chile.
Cristobal A
CA
Fuentes
Molecular Endocrinology Laboratory, Department of Endocrinology, School of Medicine, Pontificia Universidad Catolica de Chile, Santiago, Chile.
Juan E
JE
Tichauer
Molecular Endocrinology Laboratory, Department of Endocrinology, School of Medicine, Pontificia Universidad Catolica de Chile, Santiago, Chile.
Carolina
C
Valdivia
Department of Clinical Laboratories, School of Medicine, Pontificia Universidad Catolica de Chile, Santiago, Chile.
Sandra
S
Solari
Molecular Endocrinology Laboratory, Department of Endocrinology, School of Medicine, Pontificia Universidad Catolica de Chile, Santiago, Chile; Millennium Institute of Immunology and Immunotherapy, Santiago, Chile.
Carmen
C
Campino
Molecular Endocrinology Laboratory, Department of Endocrinology, School of Medicine, Pontificia Universidad Catolica de Chile, Santiago, Chile.
Alejandra
A
Tapia-Castillo
Molecular Endocrinology Laboratory, Department of Endocrinology, School of Medicine, Pontificia Universidad Catolica de Chile, Santiago, Chile; Millennium Institute of Immunology and Immunotherapy, Santiago, Chile.
Rene
R
Baudrand
Institute of Nutrition and Food Technology (INTA), Universidad de Chile, Santiago, Chile.
Pia
P
Villarroel
Institute of Nutrition and Food Technology (INTA), Universidad de Chile, Santiago, Chile.
Mariana
M
Cifuentes
Department of Physiology, Faculty of Biological Sciences, Pontificia Universidad Catolica de Chile, Santiago, Chile.
Gareth I
GI
Owen
Molecular Endocrinology Laboratory, Department of Endocrinology, School of Medicine, Pontificia Universidad Catolica de Chile, Santiago, Chile; Millennium Institute of Immunology and Immunotherapy, Santiago, Chile.
Cristian A
CA
Carvajal
Molecular Endocrinology Laboratory, Department of Endocrinology, School of Medicine, Pontificia Universidad Catolica de Chile, Santiago, Chile; Millennium Institute of Immunology and Immunotherapy, Santiago, Chile. Electronic address: cfardella@med.puc.cl.
Carlos E
CE
Fardella
D043205:11-beta-Hydroxysteroid Dehydrogenase Type 1 / Q000037:antagonists & inhibitors* / Q000737:chemistry / Q000235:genetics; D043209:11-beta-Hydroxysteroid Dehydrogenase Type 2 / Q000037:antagonists & inhibitors* / Q000737:chemistry / Q000235:genetics; D017667:Adipocytes / Q000166:cytology / Q000187:drug effects* / Q000201:enzymology; D002454:Cell Differentiation; D045744:Cell Line, Tumor; D055808:Drug Discovery; D004791:Enzyme Inhibitors / Q000737:chemistry* / Q000494:pharmacology; D015870:Gene Expression; D006801:Humans; D006854:Hydrocortisone / Q000037:antagonists & inhibitors / Q000096:biosynthesis; D007700:Kinetics; D008024:Ligands; D062105:Molecular Docking Simulation; D013329:Structure-Activity Relationship; D014584:User-Computer Interface
D004791:Enzyme Inhibitors; D008024:Ligands; D043205:11-beta-Hydroxysteroid Dehydrogenase Type 1; D043209:11-beta-Hydroxysteroid Dehydrogenase Type 2; D006854:Hydrocortisone
11β-HSD1 inhibitor; 11β-hydroxysteroid dehydrogenase type 1; HPLC-MS/MS cortisol determination; LS14 cells; Metabolic syndrome; Virtual screening