Isolation and structure elucidation of parnafungins, antifungal natural products that inhibit mRNA polyadenylation.
The Candida albicans Fitness Test, a whole-cell screening platform, was used to profile crude fermentation extracts for novel antifungal natural products with interesting mechanisms of action. An extract with intrinsic antifungal activity from the fungus Fusarium larvarum displayed a Fitness Test profile that strongly implicated mRNA processing as the molecular target responsible for inhibition of fungal growth. Isolation of the active components from this sample identified a novel class of isoxazolidinone-containing natural products, which we have named parnafungins. These natural products were isolated as an interconverting mixture of four structural- and stereoisomers. The isomerization of the parnafungins was due to a retro-Michael ring-opening and subsequent reformation of a xanthone ring system. This interconversion was blocked by methylation of an enol moiety. Structure elucidation of purified parnafungin derivatives was accomplished by X-ray crystallography and NMR analysis. The biochemical target of these natural products has been identified as the fungal polyadenosine polymerase. Parnafungins demonstrated broad spectrum antifungal activity with no observed activity against gram-positive or gram-negative bacteria. The intact isoxazolidinone ring was required for antifungal activity. In addition, the natural products were efficacious in a mouse model of disseminated candidiasis.
Parish
Craig A
CA
Natural Products Chemistry, Infectious Diseases, and Process Research, Merck Research Laboratories, Merck and Company, P.O. Box 2000, Rahway, New Jersey 07065, USA. craig_parish@merck.com
Smith
Scott K
SK
Calati
Kathleen
K
Zink
Deborah
D
Wilson
Kenneth
K
Roemer
Terry
T
Jiang
Bo
B
Xu
Deming
D
Bills
Gerald
G
Platas
Gonzalo
G
Peláez
Fernando
F
Díez
Maria Teresa
MT
Tsou
Nancy
N
McKeown
Arlene E
AE
Ball
Richard G
RG
Powles
Mary Ann
MA
Yeung
Lai
L
Liberator
Paul
P
Harris
Guy
G
D000935:Antifungal Agents / Q000737:chemistry / Q000302:isolation & purification* / Q000494:pharmacology; D002176:Candida albicans / Q000187:drug effects; D018360:Crystallography, X-Ray; D005670:Fusarium / Q000737:chemistry*; D008826:Microbial Sensitivity Tests; D008958:Models, Molecular; D019906:Nuclear Magnetic Resonance, Biomolecular; D023303:Oxazolidinones / Q000737:chemistry / Q000302:isolation & purification* / Q000494:pharmacology; D021241:Spectrometry, Mass, Electrospray Ionization; D013055:Spectrophotometry, Infrared; D013056:Spectrophotometry, Ultraviolet
D000935:Antifungal Agents; D023303:Oxazolidinones