title: Miniaturized Cultivation of Microbiota for Antimalarial Drug Discovery.
abstract: The ongoing search for effective antiplasmodial agents remains essential in the fight against malaria worldwide. Emerging parasitic drug resistance places an urgent need to explore chemotherapies with novel structures and mechanisms of action. Natural products have historically provided effective antimalarial drug scaffolds. In an effort to search nature's chemical potential for antiplasmodial agents, unconventionally sourced organisms coupled with innovative cultivation techniques were utilized. Approximately 60,000 niche microbes from various habitats (slow-growing terrestrial fungi, Antarctic microbes, and mangrove endophytes) were cultivated on a small-scale, extracted, and used in high-throughput screening to determine antimalarial activity. About 1% of crude extracts were considered active and 6% partially active (≥ 67% inhibition at 5 and 50 μg/mL, respectively). Active extracts (685) were cultivated on a large-scale, fractionated, and screened for both antimalarial activity and cytotoxicity. High interest fractions (397) with an IC50 < 1.11 μg/mL were identified and subjected to chromatographic separation for compound characterization and dereplication. Identifying active compounds with nanomolar antimalarial activity coupled with a selectivity index tenfold higher was accomplished with two of the 52 compounds isolated. This microscale, high-throughput screening project for antiplasmodial agents is discussed in the context of current natural product drug discovery efforts.
authors.affiliation: Department of Chemistry, University of South Florida, Tampa, Florida, 33620, USA.
authors.forename: Carrie
authors.initials: C
authors.lastname: Waterman
authors.affiliation: Department of Chemistry, University of South Florida, Tampa, Florida, 33620, USA.
authors.forename: Laurent
authors.initials: L
authors.lastname: Calcul
authors.affiliation: Department of Chemistry, University of South Florida, Tampa, Florida, 33620, USA.
authors.forename: Jeremy
authors.initials: J
authors.lastname: Beau
authors.affiliation: Department of Chemistry, University of South Florida, Tampa, Florida, 33620, USA.
authors.forename: Wai Sheung
authors.initials: WS
authors.lastname: Ma
authors.affiliation: Department of Chemistry, University of South Florida, Tampa, Florida, 33620, USA.
authors.forename: Matthew D
authors.initials: MD
authors.lastname: Lebar
authors.affiliation: Department of Chemistry, University of South Florida, Tampa, Florida, 33620, USA.
authors.forename: Jacqueline L
authors.initials: JL
authors.lastname: von Salm
authors.affiliation: Department of Chemistry, University of South Florida, Tampa, Florida, 33620, USA.
authors.forename: Charles
authors.initials: C
authors.lastname: Harter
authors.affiliation: Department of Global Health, University of South Florida, Tampa, Florida, 33620, USA.
authors.forename: Tina
authors.initials: T
authors.lastname: Mutka
authors.affiliation: Department of Global Health, University of South Florida, Tampa, Florida, 33620, USA.
authors.forename: Lindsay C
authors.initials: LC
authors.lastname: Morton
authors.affiliation: Department of Global Health, University of South Florida, Tampa, Florida, 33620, USA.
authors.forename: Patrick
authors.initials: P
authors.lastname: Maignan
authors.affiliation: Department of Global Health, University of South Florida, Tampa, Florida, 33620, USA.
authors.forename: Betty
authors.initials: B
authors.lastname: Barisic
authors.affiliation: Department of Global Health, University of South Florida, Tampa, Florida, 33620, USA.
authors.forename: Alberto
authors.initials: A
authors.lastname: van Olphen
authors.affiliation: Department of Global Health, University of South Florida, Tampa, Florida, 33620, USA.
authors.forename: Dennis E
authors.initials: DE
authors.lastname: Kyle
authors.affiliation: Department of Biology and Chemistry, City University of Hong Kong, Tat Chee Avenue, Kowloon, Hong Kong.
authors.forename: Lilian
authors.initials: L
authors.lastname: Vrijmoed
authors.affiliation: Institute of Marine Biology and Center of Excellence for the Oceans, National Taiwan Ocean University, Keelung, 20224, Taiwan.
authors.forename: Ka-Lai
authors.initials: KL
authors.lastname: Pang
authors.affiliation: Mycosynthetix Inc, Hillsborough, North Carolina, 27278, USA.
authors.forename: Cedric J
authors.initials: CJ
authors.lastname: Pearce
authors.affiliation: Department of Chemistry, University of South Florida, Tampa, Florida, 33620, USA.|Center for Drug Discovery and Innovation, University of South Florida, Tampa, Florida, 36612, USA.
authors.forename: Bill J
authors.initials: BJ
authors.lastname: Baker
mesh_terms: D000818:Animals; D000962:Antimalarials / Q000302:isolation & purification*; D001419:Bacteria / Q000254:growth & development*; D001431:Bacteriological Techniques / Q000379:methods*; D001681:Biological Assay; D045744:Cell Line, Tumor; D002522:Chlorocebus aethiops; D002845:Chromatography; D004285:Dogs; D055808:Drug Discovery; D004351:Drug Resistance; D005658:Fungi / Q000254:growth & development*; D006801:Humans; D020128:Inhibitory Concentration 50; D007448:Invertebrates / Q000382:microbiology; D061985:Madin Darby Canine Kidney Cells; D009682:Magnetic Resonance Spectroscopy; D008288:Malaria / Q000188:drug therapy; D064307:Microbiota; D008904:Miniaturization; D009172:Mycology / Q000379:methods*; D010936:Plant Extracts / Q000737:chemistry; D010963:Plasmodium falciparum / Q000187:drug effects; D014709:Vero Cells
chemical_list: D000962:Antimalarials; D010936:Plant Extracts
keywords: endophytes; high-throughput screening; malaria; microbe cultivation; natural products