title: Activation and inhibition of adenylyl cyclase isoforms by forskolin analogs.
abstract: Adenylyl cyclase (AC) isoforms 1 to 9 are differentially expressed in tissues and constitute an interesting drug target. ACs 1 to 8 are activated by the diterpene, forskolin (FS). It is unfortunate that there is a paucity of AC isoform-selective activators. To develop such compounds, an understanding of the structure/activity relationships of diterpenes is necessary. Therefore, we examined the effects of FS and nine FS analogs on ACs 1, 2, and 5 expressed in Spodoptera frugiperda insect cells. Diterpenes showed the highest potencies at AC1 and the lowest potencies at AC2. We identified full agonists, partial agonists, antagonists, and inverse agonists, i.e., diterpenes that reduced basal AC activity. Each AC isoform exhibited a distinct pharmacological profile. AC2 showed the highest basal activity of all AC isoforms and highest sensitivity to inverse agonistic effects of 1-deoxy-forskolin, 7-deacetyl-1,9-dideoxy-forskolin, and, particularly, BODIPY-forskolin. In contrast, BODIPY-forskolin acted as partial agonist at the other ACs. 1-Deoxy-forskolin analogs were devoid of agonistic activity at ACs but antagonized the effects of FS in a mixed competitive/noncompetitive manner. At purified catalytic AC subunits, BODIPY-forskolin acted as weak partial agonist/strong partial antagonist. Molecular modeling revealed that the BODIPY group rotates promiscuously outside of the FS-binding site. Collectively, ACs are not uniformly activated and inhibited by FS and FS analogs, demonstrating the feasibility to design isoform-selective FS analogs. The two- and multiple-state models, originally developed to conceptualize ligand effects at G-protein-coupled receptors, can be applied to ACs to explain certain experimental data.
authors.affiliation: Department of Pharmacology and Toxicology, University of Regensburg, Universitätsstrasse 31, D-93053 Regensburg, Germany.
authors.forename: Cibele
authors.initials: C
authors.lastname: Pinto
authors.forename: Dan
authors.initials: D
authors.lastname: Papa
authors.forename: Melanie
authors.initials: M
authors.lastname: Hübner
authors.forename: Tung-Chung
authors.initials: TC
authors.lastname: Mou
authors.forename: Gerald H
authors.initials: GH
authors.lastname: Lushington
authors.forename: Roland
authors.initials: R
authors.lastname: Seifert
mesh_terms: D000067956:Adenylyl Cyclase Inhibitors; D000262:Adenylyl Cyclases / Q000187:drug effects*; D000818:Animals; D002460:Cell Line; D005576:Colforsin / Q000737:chemistry / Q000494:pharmacology*; D004224:Diterpenes; D004285:Dogs; D015195:Drug Design; D004789:Enzyme Activation / Q000187:drug effects; D004791:Enzyme Inhibitors / Q000494:pharmacology; D007527:Isoenzymes / Q000187:drug effects; D051379:Mice; D013329:Structure-Activity Relationship; D014162:Transfection
chemical_list: D000067956:Adenylyl Cyclase Inhibitors; D004224:Diterpenes; D004791:Enzyme Inhibitors; D007527:Isoenzymes; D005576:Colforsin; D000262:Adenylyl Cyclases; C100622:adenylyl cyclase 2