title: Antifungal activity and cytotoxicity of extracts and triterpenoid saponins obtained from the aerial parts of Anagallis arvensis L.
abstract: BACKGROUND
  Anagallis arvensis L. (Primulaceae) is used in argentinean northwestern traditional medicine to treat fungal infections. We are reporting the isolation and identification of compounds with antifungal activity against human pathogenic yeast Candida albicans, and toxicity evaluation.
  
  
  OBJECTIVE
  to study the antifungal activity of extracts and purified compounds obtained form A. arvensis aerial parts, alone and in combinations with fluconazole (FLU), and to study the toxicity of the active compounds.
  
  
  METHODS
  Disk diffusion assays were used to perform an activity-guided isolation of antifungal compounds from the aerial parts of A. arvensis. Broth dilution checkerboard and viable cell count assays were employed to determine the effects of samples and combinations of FLU + samples against Candida albicans. The chemical structures of active compounds were elucidated by spectroscopic analysis. Genotoxic and haemolytic effects of the isolated compounds were determined.
  
  
  RESULTS
  Four triterpenoid saponins (1-4) were identified. Anagallisin C (AnC), exerted the highest inhibitory activity among the assayed compounds against C. albicans reference strain (ATCC 10231), with MIC-0 =1µg/mL. The Fractional Inhibitory Concentration Index (FICI=0.129) indicated a synergistic effect between AnC (0.125µg/mL) and FLU (0.031µg/mL) against C. albicans ATCC 10231. AnC inhibited C. albicans 12-99 FLU resistant strain (MIC-0 =1µg/mL), and the FICI=0.188 indicated a synergistic effect between AnC (0.125µg/mL) and fluconazole (16µg/mL). The combination AnC+ FLU exerted fungicidal activity against both C. albicans strains. AnC exerted inhibitory activity against C. albicans ATCC 10231 sessile cells (MIC50=0.5µg/mL and MIC80=1µg/mL) and against C. albicans 12-99 sessile cells (MIC50=0.75µg/mL and MIC80=1.25µg/mL). AnC exerted haemolytic effect against human red blood cells at 15µg/mL and did not exerted genotoxic effect on Bacillus subtilis rec strains.
  
  
  CONCLUSIONS
  The antifungal activity and lack of genotoxic effects of AnC give support to the traditional use of A. arvensis as antifungal and makes AnC a compound of interest to expand the available antifungal drugs.
authors.affiliation: Cátedra de Fitoquímica, Instituto de Estudios Farmacológicos "Dr. A.R. Sampietro", Facultad de Bioquímica, Química y Farmacia, Universidad Nacional de Tucumán, Ayacucho 471, T4000INI San Miguel de Tucumán, Tucumán, Argentina; Consejo Nacional de Investigaciones Científicas y Técnicas (CONICET), Godoy Cruz 2290, C1425FQB Ciudad Autónoma de Buenos Aires, Argentina. Electronic address: jrsrody@yahoo.com.
authors.forename: José R
authors.initials: JR
authors.lastname: Soberón
authors.affiliation: Cátedra de Fitoquímica, Instituto de Estudios Farmacológicos "Dr. A.R. Sampietro", Facultad de Bioquímica, Química y Farmacia, Universidad Nacional de Tucumán, Ayacucho 471, T4000INI San Miguel de Tucumán, Tucumán, Argentina; Consejo Nacional de Investigaciones Científicas y Técnicas (CONICET), Godoy Cruz 2290, C1425FQB Ciudad Autónoma de Buenos Aires, Argentina.
authors.forename: Melina A
authors.initials: MA
authors.lastname: Sgariglia
authors.affiliation: Cátedra de Fitoquímica, Instituto de Estudios Farmacológicos "Dr. A.R. Sampietro", Facultad de Bioquímica, Química y Farmacia, Universidad Nacional de Tucumán, Ayacucho 471, T4000INI San Miguel de Tucumán, Tucumán, Argentina.
authors.forename: Ana C
authors.initials: AC
authors.lastname: Pastoriza
authors.affiliation: Cátedra de Fitoquímica, Instituto de Estudios Farmacológicos "Dr. A.R. Sampietro", Facultad de Bioquímica, Química y Farmacia, Universidad Nacional de Tucumán, Ayacucho 471, T4000INI San Miguel de Tucumán, Tucumán, Argentina.
authors.forename: Estela M
authors.initials: EM
authors.lastname: Soruco
authors.affiliation: Instituto de Química Rosario, UNR, CONICET, Suipacha 531, S2002LRK Rosario, Argentina.
authors.forename: Sebastián N
authors.initials: SN
authors.lastname: Jäger
authors.affiliation: Instituto de Química Rosario, UNR, CONICET, Suipacha 531, S2002LRK Rosario, Argentina; Facultad de Ciencias Bioquímicas y Farmacéuticas, Universidad Nacional de Rosario, Suipacha 531, S2002LRK Rosario, Argentina.
authors.forename: Guillermo R
authors.initials: GR
authors.lastname: Labadie
authors.affiliation: Cátedra de Fitoquímica, Instituto de Estudios Farmacológicos "Dr. A.R. Sampietro", Facultad de Bioquímica, Química y Farmacia, Universidad Nacional de Tucumán, Ayacucho 471, T4000INI San Miguel de Tucumán, Tucumán, Argentina; Consejo Nacional de Investigaciones Científicas y Técnicas (CONICET), Godoy Cruz 2290, C1425FQB Ciudad Autónoma de Buenos Aires, Argentina.
authors.forename: Diego A
authors.initials: DA
authors.lastname: Sampietro
authors.affiliation: Consejo Nacional de Investigaciones Científicas y Técnicas (CONICET), Godoy Cruz 2290, C1425FQB Ciudad Autónoma de Buenos Aires, Argentina.
authors.forename: Marta A
authors.initials: MA
authors.lastname: Vattuone
mesh_terms: D031823:Anagallis / Q000737:chemistry*; D000935:Antifungal Agents / Q000302:isolation & purification / Q000494:pharmacology* / Q000633:toxicity; D002176:Candida albicans / Q000187:drug effects; D052978:Disk Diffusion Antimicrobial Tests; D004357:Drug Synergism; D015725:Fluconazole / Q000008:administration & dosage / Q000494:pharmacology; D006461:Hemolysis / Q000187:drug effects; D006801:Humans; D008519:Medicine, Traditional; D009152:Mutagenicity Tests; D035261:Plant Components, Aerial; D010936:Plant Extracts / Q000737:chemistry / Q000494:pharmacology* / Q000633:toxicity; D012503:Saponins / Q000302:isolation & purification / Q000494:pharmacology* / Q000633:toxicity; D014315:Triterpenes / Q000302:isolation & purification / Q000494:pharmacology / Q000633:toxicity
chemical_list: D000935:Antifungal Agents; D010936:Plant Extracts; D012503:Saponins; D014315:Triterpenes; D015725:Fluconazole
keywords: Anagallis arvensis; Anagallisin C; Antifungal activity; Biofilm; Candida albicans; Synergistic effect