title: Discovery of Tricyclic Clerodane Diterpenes as Sarco/Endoplasmic Reticulum Ca(2+)-ATPase Inhibitors and Structure-Activity Relationships.
abstract: Tricyclic clerodane diterpenes (TCDs) are natural compounds that often show potent cytotoxicity for cancer cells, but their mode of action remains elusive. A computationally based similarity search (CDRUG), combined with principal component analysis (ChemGPS-NP) and docking calculations (GOLD 5.2), suggested TCDs to be inhibitors of the sarco/endoplasmic reticulum Ca(2+)-ATPase (SERCA) pump, which is also the target of the sesquiterpene lactone thapsigargin. Biochemical studies were performed with 11 TCDs on purified rabbit skeletal muscle sarcoplasmic reticulum membranes, which are highly enriched with the SERCA1a isoform. Casearborin D (2) exhibited the highest affinity, with a KD value of 2 μM and giving rise to complete inhibition of SERCA1a activity. Structure-activity relationships revealed that functionalization of two acyl side chains (R1 and R4) and the hydrophobicity imparted by the aliphatic chain at C-9, as well as a C-3,C-4 double bond, play crucial roles for inhibitory activity. Docking studies also suggested that hydrophobic interactions in the binding site, especially with Phe256 and Phe834, may be important for a strong inhibitory activity of the TCDs. In conclusion, a novel class of SERCA inhibitory compounds is presented.
authors.affiliation: †Department of Pharmaceutical Biology and Biotechnology, Albert Ludwigs University Freiburg, Stefan-Meier-Strasse 19, 79104 Freiburg, Germany.|‡Spemann Graduate School of Biology and Medicine (SGBM), Albert Ludwigs University Freiburg, Albertstrasse 19a, 79104 Freiburg, Germany.|§Faculty of Chemistry and Pharmacy, Albert Ludwigs University Freiburg, Albertstrasse 25, 79104 Freiburg, Germany.
authors.forename: Christian
authors.initials: C
authors.lastname: De Ford
authors.affiliation: #Escuela de Química and CIPRONA, Universidad de Costa Rica, 2060 San José, Costa Rica.
authors.forename: Carlos
authors.initials: C
authors.lastname: Calderón
authors.affiliation: ∥Department of Biomedicine, Aarhus University, Ole Worms Allé 3, DK-8000 Aarhus C, Denmark.|⊥Centre for Membrane Pumps in Cells and Disease (PUMPkin), National Research Foundation, Aarhus, Denmark.
authors.forename: Pankaj
authors.initials: P
authors.lastname: Sehgal
authors.affiliation: ∥Department of Biomedicine, Aarhus University, Ole Worms Allé 3, DK-8000 Aarhus C, Denmark.|⊥Centre for Membrane Pumps in Cells and Disease (PUMPkin), National Research Foundation, Aarhus, Denmark.
authors.forename: Natalya U
authors.initials: NU
authors.lastname: Fedosova
authors.affiliation: #Escuela de Química and CIPRONA, Universidad de Costa Rica, 2060 San José, Costa Rica.
authors.forename: Renato
authors.initials: R
authors.lastname: Murillo
authors.affiliation: ∥Department of Biomedicine, Aarhus University, Ole Worms Allé 3, DK-8000 Aarhus C, Denmark.|⊥Centre for Membrane Pumps in Cells and Disease (PUMPkin), National Research Foundation, Aarhus, Denmark.
authors.forename: Claus
authors.initials: C
authors.lastname: Olesen
authors.affiliation: ∥Department of Biomedicine, Aarhus University, Ole Worms Allé 3, DK-8000 Aarhus C, Denmark.|⊥Centre for Membrane Pumps in Cells and Disease (PUMPkin), National Research Foundation, Aarhus, Denmark.
authors.forename: Poul
authors.initials: P
authors.lastname: Nissen
authors.affiliation: ∥Department of Biomedicine, Aarhus University, Ole Worms Allé 3, DK-8000 Aarhus C, Denmark.|⊥Centre for Membrane Pumps in Cells and Disease (PUMPkin), National Research Foundation, Aarhus, Denmark.
authors.forename: Jesper V
authors.initials: JV
authors.lastname: Møller
authors.affiliation: †Department of Pharmaceutical Biology and Biotechnology, Albert Ludwigs University Freiburg, Stefan-Meier-Strasse 19, 79104 Freiburg, Germany.|‡Spemann Graduate School of Biology and Medicine (SGBM), Albert Ludwigs University Freiburg, Albertstrasse 19a, 79104 Freiburg, Germany.|§Faculty of Chemistry and Pharmacy, Albert Ludwigs University Freiburg, Albertstrasse 25, 79104 Freiburg, Germany.
authors.forename: Irmgard
authors.initials: I
authors.lastname: Merfort
mesh_terms: D000251:Adenosine Triphosphatases / Q000037:antagonists & inhibitors*; D000592:Amino Acid Metabolism, Inborn Errors; D000818:Animals; D001665:Binding Sites; D045785:Diterpenes, Clerodane / Q000737:chemistry / Q000302:isolation & purification* / Q000494:pharmacology*; D004354:Drug Screening Assays, Antitumor; D004721:Endoplasmic Reticulum / Q000378:metabolism; D006801:Humans; D028361:Mitochondrial Diseases; D015394:Molecular Structure; D019906:Nuclear Magnetic Resonance, Biomolecular; D011817:Rabbits; D012519:Sarcoplasmic Reticulum / Q000378:metabolism; D050892:Sarcosine Dehydrogenase / Q000172:deficiency; D013329:Structure-Activity Relationship; D019284:Thapsigargin / Q000494:pharmacology
chemical_list: D045785:Diterpenes, Clerodane; C000601077:casearborin D; D019284:Thapsigargin; D050892:Sarcosine Dehydrogenase; D000251:Adenosine Triphosphatases