title: Semi-synthetic ecdysteroids as gene-switch actuators: synthesis, structure-activity relationships, and prospective ADME properties.
abstract: The ligand-inducible, ecdysteroid receptor (EcR) gene-expression system can add critical control features to protein expression in cell and gene therapy. However, potent natural ecdysteroids possess absorption, distribution, metabolism and excretion (ADME) properties that have not been optimised for use as gene-switch actuators in vivo. Herein we report the first systematic synthetic exploration of ecdysteroids toward modulation of gene-switch potency. Twenty-three semi-synthetic O-alkyl ecdysteroids were assayed in both a natural insect system (Drosophila B(II) cells) and engineered gene-switch systems in mammalian cells using Drosophila melanogaster, Choristoneura fumiferana, and Aedes aegypti EcRs. Gene-switch potency is maintained, or even enhanced, for ecdysteroids methylated at the 22-position in favourable cases. Furthermore, trends toward lower solubility, higher permeability, and higher blood-brain barrier penetration are supported by predicted ADME properties, calculated using the membrane-interaction (MI)-QSAR methodology. The structure-activity relationship (SAR) of alkylated ecdysteroids indicates that 22-OH is an H-bond acceptor, 25-OH is most likely an H-bond donor, and 2-OH and 3-OH are donors and/or acceptors in network with each other, and with the EcR. The strategy of alkylation points the way to improved ecdysteroidal actuators for switch-activated gene therapy.
authors.affiliation: Department of Biological Sciences, University of Exeter, Prince of Wales Road, EX4 4PS, Exeter, UK. slapenna@gmail.com
authors.forename: Silvia
authors.initials: S
authors.lastname: Lapenna
authors.forename: Laurence
authors.initials: L
authors.lastname: Dinan
authors.forename: Jennifer
authors.initials: J
authors.lastname: Friz
authors.forename: Anton J
authors.initials: AJ
authors.lastname: Hopfinger
authors.forename: Jianzhong
authors.initials: J
authors.lastname: Liu
authors.forename: Robert E
authors.initials: RE
authors.lastname: Hormann
mesh_terms: D016475:3T3 Cells; D000818:Animals; D018938:Caco-2 Cells; D002478:Cells, Cultured; D004331:Drosophila melanogaster / Q000378:metabolism; D015195:Drug Design; D026461:Ecdysteroids / Q000138:chemical synthesis / Q000737:chemistry* / Q000494:pharmacology*; D015870:Gene Expression / Q000187:drug effects*; D006801:Humans; D051379:Mice; D011987:Receptors, Steroid / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D013329:Structure-Activity Relationship
chemical_list: D026461:Ecdysteroids; D011987:Receptors, Steroid; C034808:ecdysteroid receptor