title: Benzimidazole derivative small-molecule 991 enhances AMPK activity and glucose uptake induced by AICAR or contraction in skeletal muscle.
abstract: AMP-activated protein kinase (AMPK) plays diverse roles and coordinates complex metabolic pathways for maintenance of energy homeostasis. This could be explained by the fact that AMPK exists as multiple heterotrimer complexes comprising a catalytic α-subunit (α1 and α2) and regulatory β (β1 and β2)- and γ (γ1, γ2, γ3)-subunits, which are uniquely distributed across different cell types. There has been keen interest in developing specific and isoform-selective AMPK-activating drugs for therapeutic use and also as research tools. Moreover, establishing ways of enhancing cellular AMPK activity would be beneficial for both purposes. Here, we investigated if a recently described potent AMPK activator called 991, in combination with the commonly used activator 5-aminoimidazole-4-carboxamide riboside or contraction, further enhances AMPK activity and glucose transport in mouse skeletal muscle ex vivo. Given that the γ3-subunit is exclusively expressed in skeletal muscle and has been implicated in contraction-induced glucose transport, we measured the activity of AMPKγ3 as well as ubiquitously expressed γ1-containing complexes. We initially validated the specificity of the antibodies for the assessment of isoform-specific AMPK activity using AMPK-deficient mouse models. We observed that a low dose of 991 (5 μM) stimulated a modest or negligible activity of both γ1- and γ3-containing AMPK complexes. Strikingly, dual treatment with 991 and 5-aminoimidazole-4-carboxamide riboside or 991 and contraction profoundly enhanced AMPKγ1/γ3 complex activation and glucose transport compared with any of the single treatments. The study demonstrates the utility of a dual activator approach to achieve a greater activation of AMPK and downstream physiological responses in various cell types, including skeletal muscle.
authors: Nestlé Institute of Health Sciences SA, EPFL Innovation Park, Lausanne, Switzerland.
authors: Laurent
authors: L
authors: Bultot
authors: Department of Nutrition, Exercise and Sports, Faculty of Science, University of Copenhagen, Copenhagen, Denmark.
authors: Thomas E
authors: TE
authors: Jensen
authors: Université catholique de Louvain and de Duve Institute, Brussels, Belgium.
authors: Yu-Chiang
authors: YC
authors: Lai
authors: Department of Nutrition, Exercise and Sports, Faculty of Science, University of Copenhagen, Copenhagen, Denmark.
authors: Agnete L B
authors: AL
authors: Madsen
authors: Nestlé Institute of Health Sciences SA, EPFL Innovation Park, Lausanne, Switzerland; School of Life Sciences, Ecole Polytechnique Fédérale de Lausanne, Lausanne, Switzerland.
authors: Caterina
authors: C
authors: Collodet
authors: Université catholique de Louvain and de Duve Institute, Brussels, Belgium.
authors: Samanta
authors: S
authors: Kviklyte
authors: Nestlé Institute of Health Sciences SA, EPFL Innovation Park, Lausanne, Switzerland.
authors: Maria
authors: M
authors: Deak
authors: Experimental Therapeutics and Division of Cardiovascular Medicine, Radcliffe Department of Medicine, University of Oxford, Oxford, United Kingdom.
authors: Arash
authors: A
authors: Yavari
authors: U1016, Institut National de la Santé et de la Recherche Médicale, Institut Cochin, Paris, France; UMR8104, Centre National de la Recherche Scientifique, Paris, France; and Université Paris Descartes, Paris, France.
authors: Marc
authors: M
authors: Foretz
authors: Experimental Therapeutics and Division of Cardiovascular Medicine, Radcliffe Department of Medicine, University of Oxford, Oxford, United Kingdom.
authors: Sahar
authors: S
authors: Ghaffari
authors: Experimental Therapeutics and Division of Cardiovascular Medicine, Radcliffe Department of Medicine, University of Oxford, Oxford, United Kingdom.
authors: Mohamed
authors: M
authors: Bellahcene
authors: Experimental Therapeutics and Division of Cardiovascular Medicine, Radcliffe Department of Medicine, University of Oxford, Oxford, United Kingdom.
authors: Houman
authors: H
authors: Ashrafian
authors: Université catholique de Louvain and de Duve Institute, Brussels, Belgium.
authors: Mark H
authors: MH
authors: Rider
authors: Department of Nutrition, Exercise and Sports, Faculty of Science, University of Copenhagen, Copenhagen, Denmark.
authors: Erik A
authors: EA
authors: Richter
authors: Nestlé Institute of Health Sciences SA, EPFL Innovation Park, Lausanne, Switzerland; School of Life Sciences, Ecole Polytechnique Fédérale de Lausanne, Lausanne, Switzerland; Kei.Sakamoto@rd.nestle.com.
authors: Kei
authors: K
authors: Sakamoto
mesh_terms: D055372:AMP-Activated Protein Kinases / Q000187:drug effects / Q000378:metabolism*; D000620:Aminoimidazole Carboxamide / Q000031:analogs & derivatives* / Q000494:pharmacology; D000818:Animals; D019138:Antibodies, Blocking / Q000494:pharmacology; D001562:Benzimidazoles / Q000494:pharmacology*; D001565:Benzoates / Q000494:pharmacology*; D020536:Enzyme Activators / Q000494:pharmacology*; D005947:Glucose / Q000378:metabolism*; D006801:Humans; D007004:Hypoglycemic Agents / Q000494:pharmacology*; D066298:In Vitro Techniques; D007527:Isoenzymes; D051379:Mice; D018345:Mice, Knockout; D009119:Muscle Contraction / Q000187:drug effects; D018482:Muscle, Skeletal / Q000187:drug effects*; D012265:Ribonucleotides / Q000494:pharmacology*
chemical_list: C000620824:5-((6-chloro-5-(1-methylindol-5-yl)-1H-benzimidazol-2-yl)oxy)-2-methylbenzoic acid; D019138:Antibodies, Blocking; D001562:Benzimidazoles; D001565:Benzoates; D020536:Enzyme Activators; D007004:Hypoglycemic Agents; D007527:Isoenzymes; D012265:Ribonucleotides; D000620:Aminoimidazole Carboxamide; D055372:AMP-Activated Protein Kinases; C031143:AICA ribonucleotide; D005947:Glucose
keywords: 5-aminoimidazole-4-carboxamide riboside; 991; A769662; AMP-activated protein kinase; LKB1; compound 13; ex229