Figure 3

Figure 3

Bioactivity-guided identification of the active compounds of antiseizure hit SK0107. (A) Aspergillus insuetus IBT 28443 cultivated on czapek yeast extract agar (CYA) media for 9 days in the dark at 25 °C. Base peak chromatogram (BPC) of the most bioactive fraction (SK1312) from first reversed-phase fractionation in positive electrospray ionization mode (ESI⁺). ESI⁺ BPC chromatograms of the two most bioactive fractions (SK1414 and SK1415) from the second reversed-phase fractionation. UV/Vis and HRMS spectra for TMC-120A (II) and TMC-120B (III). (B–D) Antiseizure activity of SK1312 (n = 23–24 replicate wells per condition) (B), SK1414 (n = 10–11 replicate wells per condition) (C), and SK1415 (n = 22 replicate wells per condition) (D) in the zebrafish pentylenetetrazole (PTZ) seizure model after 2 h of incubation at their maximum tolerated concentration (MTC), MTC/2, and MTC/4. PTZ- induced seizure-like behavior is expressed as mean actinteg units per 5 min (±SEM) during the 30-min recording period. (B,D) Data are pooled from two independent experiments with each 11–12 replicate wells per condition. (C) Data are from a single experiment with 10–11 replicate wells per condition. (B–D) Statistical analysis: one-way ANOVA with Dunnett’s multiple comparison test for comparison of sample + PTZ groups with vehicle (VHC) + PTZ control group, Kruskal–Wallis test with Dunn’s multiple comparison test (data did not pass the Shapiro–Wilk normality test) for comparison of sample + VHC groups with VHC + VHC control group (GraphPad Prism 5, San Diego, CA, USA). Significance levels: * p ≤ 0.05; ** p ≤ 0.01; *** p ≤ 0.001.