Figure 1

Bioactivity-guided identification of the active compound halimide of antiseizure hit SK0107. (A) Aspergillus insuetus IBT 28443 cultivated on czapek yeast extract agar (CYA) and yeast extract sucrose agar (YES) media for 9 days at 25 °C in the dark. Base peak chromatograms (BPC) of the crude extract and bioactive fraction SK0107 in positive electrospray ionization mode (ESI+). (B) Aspergillus insuetus IBT 28443 cultivated on CYA media for 9 days at 25 °C in the dark. ESI+ BPC chromatograms of the most bioactive fraction (SK1312) from first reversed-phase fractionation and of the two most bioactive fractions (SK1414 and SK1415) from the second reversed-phase fractionation. UV/Vis and HRMS spectra shown for halimide (I). TMC-120A (II) and TMC-120B (III) peaks are also indicated. (C–F) Antiseizure activity of SK0107 (means are pooled from three independent experiments with 12 replicate wells per condition each) (C), SK1312 (n = 23–24 replicate wells per condition) (D), SK1414 (n = 10–11 replicate wells per condition) (E), and SK1415 (n = 22 replicate wells per condition) (F) in the zebrafish pentylenetetrazole (PTZ) seizure model after 2 h of treatment. Behavioral data is expressed in mean actinteg units per 5 min (±SEM) during the 30 min recording period. Statistical analysis: (C–F) one-way ANOVA with Dunnett’s multiple comparison test (GraphPad Prism 5, San Diego, CA, USA). Significance levels: * p ≤ 0.05; ** p ≤ 0.01; *** p ≤ 0.001. Abbreviation: vehicle: VHC.