title: Synthesis and in vitro activity of 3 beta-substituted-3 alpha-hydroxypregnan-20-ones: allosteric modulators of the GABAA receptor.
abstract: Two naturally occurring metabolites of progesterone, 3 alpha-hydroxy-5 alpha- and 5 beta-pregnan-20-one (1 and 2), are potent allosteric modulators of the GABAA receptor. Their therapeutic potential as anxiolytics, anticonvulsants, and sedative/hypnotics is limited by rapid metabolism. To avoid these shortcomings, a series of 3 beta-substituted derivatives of 1 and 2 was prepared. Small lipophilic groups generally maintain potency in both the 5 alpha- and 5 beta-series as determined by inhibition of [35S]TBPS binding. In the 5 alpha-series, 3 beta-ethyl, -propyl, -trifluoromethyl and -(benzyloxy)methyl, as well as substituents of the form 3 beta-XCH2, where X is Cl, Br, or I or contains unsaturation, show limited efficacy in inhibiting [35S]TBPS binding. In the 5 beta-series, the unsubstituted parent 2 is a two-component inhibitor, whereas all of the 3 beta-substituted derivatives of 2 inhibit TBPS via a single class of binding sites. In addition, all of the 3-substituted 5 beta-sterols tested are full inhibitors of [35S]TBPS binding. Electrophysiological measurements using alpha 1 beta 2 gamma 2L receptors expressed in oocytes show that 3 beta-methyl- and 3 beta-(azidomethyl)-3 alpha-hydroxy-5 alpha-pregnan-20-one (6 and 22, respectively) are potent full efficacy modulators and that 3 alpha-hydroxy-3 beta-(trifluoromethyl)-5 alpha-pregnan -20-one (24) is a low-efficacy modulator, confirming the results obtained from [35S]TBPS binding. These results indicate that modification of the 3 beta-position in 1 and 2 maintains activity at the neuroactive steroid site on the GABAA receptor. In animal studies, compound 6 (CCD 1042) is an orally active anticonvulsant, while the naturally occurring progesterone metabolites 1 and 2 are inactive when administered orally, suggesting that 3 beta-substitution slows metabolism of the 3-hydroxyl, resulting in orally bioavailable steroid modulators of the GABAA receptor.
authors.affiliation: CoCensys, Inc., Irvine, California 92618, USA.
authors.forename: D J
authors.initials: DJ
authors.lastname: Hogenkamp
authors.forename: S H
authors.initials: SH
authors.lastname: Tahir
authors.forename: J E
authors.initials: JE
authors.lastname: Hawkinson
authors.forename: R B
authors.initials: RB
authors.lastname: Upasani
authors.forename: M
authors.initials: M
authors.lastname: Alauddin
authors.forename: C L
authors.initials: CL
authors.lastname: Kimbrough
authors.forename: M
authors.initials: M
authors.lastname: Acosta-Burruel
authors.forename: E R
authors.initials: ER
authors.lastname: Whittemore
authors.forename: R M
authors.initials: RM
authors.lastname: Woodward
authors.forename: N C
authors.initials: NC
authors.lastname: Lan
authors.forename: K W
authors.initials: KW
authors.lastname: Gee
authors.forename: M B
authors.initials: MB
authors.lastname: Bolger
mesh_terms: D000818:Animals; D014151:Anti-Anxiety Agents / Q000737:chemistry* / Q000378:metabolism; D019086:Bridged Bicyclo Compounds, Heterocyclic / Q000378:metabolism; D003292:Convulsants / Q000378:metabolism; D003900:Desoxycorticosterone / Q000031:analogs & derivatives* / Q000737:chemistry / Q000378:metabolism; D004594:Electrophysiology; D005260:Female; D066298:In Vitro Techniques; D008958:Models, Molecular; D009865:Oocytes / Q000378:metabolism; D051381:Rats; D011963:Receptors, GABA-A / Q000378:metabolism*; D014981:Xenopus
chemical_list: D014151:Anti-Anxiety Agents; D019086:Bridged Bicyclo Compounds, Heterocyclic; D003292:Convulsants; D011963:Receptors, GABA-A; D003900:Desoxycorticosterone; C009413:tetrahydrodeoxycorticosterone; C037476:tert-butylbicyclophosphorothionate