title: Rustmicin, a potent antifungal agent, inhibits sphingolipid synthesis at inositol phosphoceramide synthase.
abstract: Rustmicin is a 14-membered macrolide previously identified as an inhibitor of plant pathogenic fungi by a mechanism that was not defined. We discovered that rustmicin inhibits inositol phosphoceramide synthase, resulting in the accumulation of ceramide and the loss of all of the complex sphingolipids. Rustmicin has potent fungicidal activity against clinically important human pathogens that is correlated with its sphingolipid inhibition. It is especially potent against Cryptococcus neoformans, where it inhibits growth and sphingolipid synthesis at concentrations <1 ng/ml and inhibits the enzyme with an IC50 of 70 pM. This inhibition of the membrane-bound enzyme is reversible; moreover, rustmicin is nearly equipotent against the solubilized enzyme. Rustmicin was efficacious in a mouse model for cryptococcosis, but it was less active than predicted from its in vitro potency against this pathogen. Stability and drug efflux were identified as two factors limiting rustmicin's activity. In the presence of serum, rustmicin rapidly epimerizes at the C-2 position and is converted to a gamma-lactone, a product that is devoid of activity. Rustmicin was also found to be a remarkably good substrate for the Saccharomyces cerevisiae multidrug efflux pump encoded by PDR5.
authors.affiliation: Department of Biochemistry, Merck Research Laboratories, Rahway, New Jersey 07065, USA. suzanne_mandala@merck.com
authors.forename: S M
authors.initials: SM
authors.lastname: Mandala
authors.forename: R A
authors.initials: RA
authors.lastname: Thornton
authors.forename: J
authors.initials: J
authors.lastname: Milligan
authors.forename: M
authors.initials: M
authors.lastname: Rosenbach
authors.forename: M
authors.initials: M
authors.lastname: Garcia-Calvo
authors.forename: H G
authors.initials: HG
authors.lastname: Bull
authors.forename: G
authors.initials: G
authors.lastname: Harris
authors.forename: G K
authors.initials: GK
authors.lastname: Abruzzo
authors.forename: A M
authors.initials: AM
authors.lastname: Flattery
authors.forename: C J
authors.initials: CJ
authors.lastname: Gill
authors.forename: K
authors.initials: K
authors.lastname: Bartizal
authors.forename: S
authors.initials: S
authors.lastname: Dreikorn
authors.forename: M B
authors.initials: MB
authors.lastname: Kurtz
mesh_terms: D000818:Animals; D000935:Antifungal Agents / Q000494:pharmacology; D002455:Cell Division / Q000187:drug effects; D003453:Cryptococcosis / Q000188:drug therapy; D003455:Cryptococcus neoformans / Q000187:drug effects / Q000472:pathogenicity; D004791:Enzyme Inhibitors / Q000494:pharmacology; D005656:Fungal Proteins / Q000378:metabolism; D005658:Fungi / Q000201:enzymology / Q000472:pathogenicity; D006028:Glycosphingolipids / Q000096:biosynthesis*; D006602:Hexosyltransferases / Q000037:antagonists & inhibitors*; D007294:Inositol / Q000378:metabolism; D007783:Lactones / Q000378:metabolism / Q000494:pharmacology; D008565:Membrane Proteins / Q000378:metabolism; D051379:Mice; D008815:Mice, Inbred Strains; D015394:Molecular Structure; D012441:Saccharomyces cerevisiae / Q000187:drug effects; D013107:Sphingolipids / Q000096:biosynthesis*
chemical_list: D000935:Antifungal Agents; D004791:Enzyme Inhibitors; D005656:Fungal Proteins; D006028:Glycosphingolipids; D007783:Lactones; D008565:Membrane Proteins; D013107:Sphingolipids; C025524:inositolphosphoceramides; C047784:rustmicin; D007294:Inositol; D006602:Hexosyltransferases; C088714:phosphatidylinositol-ceramide phosphoinositol transferase