*3.3. Fermentation, Extraction, and Isolation*

The fungus was incubated on potato dextrose agar (PDA) medium at 28 ◦C for approximately 2–3 days; then it was cut into three agar pieces (nearly the size of 0.5 × 0.5 × 0.5 cm) and transferred into a 500 mL Erlenmeyer flask, containing 200 mL of potato dextrose broth (PDB). The flasks were cultured for 2 days at 28 ◦C on a rotary shaker at 180 rpm for inoculation. The seed cultures were added to the 200 × 1 L flasks containing rice medium (110 g rice, 120 mL deionized water), which was previously sterilized at 121 ◦C for 25 min. All flasks were incubated at 28 ◦C for four weeks.

Following incubation, the solid rice cultures were extracted three times by EtOAc to give a crude extract (489.0 g); the crude extract was suspended in water and then partitioned with EtOAc to give an EtOAc soluble fraction (185.2 g) after the solvent was removed to dryness under reduced pressure. The EtOAc fraction was further separated on macroporous adsorbent resin with a stepped gradient elution with EtOH-H2O (30, 50, 70 and 100%). The 50% fraction was sequentially separated by silica gel with petroleum ether-EtOAc (5:1 to 0:1) to give four subfractions (A**–**D) using the TLC. The subfraction B (9.0 g) was sequentially loaded onto silica gel CC (petroleum ether-EtOAc, 5:1) and preparative HPLC (MeCN-H2O, 50:50, 10.0 mL/min) to yield compounds **1** (10.4 mg, *t*<sup>R</sup> 34.9 min), **2** (6.4 mg, *t*<sup>R</sup> 27.1 min), **4** (5.9 mg, *t*<sup>R</sup> 33.5 min), and **6** (8.7 mg, *t*<sup>R</sup> 23.2 min). Subfraction C (15.0 g) was further separated by an ODS column (MeCN-H2O, 40:60) and repeated preparative HPLC with MeCN-H2O (50:50, 10 mL/min) to give compounds **3** (4.6 mg, *t*<sup>R</sup> 8.8 min) and **5** (4.7 mg, *t*<sup>R</sup> 17.1 min).

Aspamide A (**1**): yellowish powder; [α] 25 <sup>D</sup> + 120.0 (*c* 0.05, MeOH); ECD (5 mg/L, MeOH) λmax (Δε) 212 (92.18), 245 (−41.48) 335 (19.29) nm; UV (MeOH) λmax (logε) 200 (6.13), 224 (6.13), 284 (5.46), 341 (5.68) nm; 1H and 13C NMR (DMSO-*d*6), see Table 1; positive HR-ESI-MS *m*/*z* 394.2117 [M + H] <sup>+</sup>, (calcd. for C23H28N3O3, 394.2125).

Aspamide B (**2**): yellowish powder; [α] 25 <sup>D</sup> + 112.0 (*c* 0.05, MeOH); ECD (5 mg/L, MeOH) λmax (Δε) 212 (83.31), 261 (−37.91), 340 (20.41) nm; UV (MeOH) λmax (logε) 200 (6.18), 224 (6.18), 283 (5.45), 345 (5.73) nm; 1H and 13C NMR (DMSO-*d6*), see Table 1; positive HR-ESI-MS *m*/*z* 394.2120 [M + H]+, (calcd. for C23H28N3O3, 394.2125).

Aspamide C (**3**): yellowish powder; [α] 25 <sup>D</sup> + 125.0 (*c* 0.05, MeOH); ECD (5 mg/L, MeOH) λmax (Δε) 211 (103.75), 255 (−44.38), 334 (24.79) nm; UV (MeOH) λmax (logε) 200 (6.20), 224 (6.19), 284 (5.56), 337 (5.75) nm; 1H and 13C NMR (DMSO-*d6*), see Table 1; positive HR-ESI-MS *m*/*z* 366.1807 [M + H]+, (calcd. for C21H24N3O3, 366.1812).

(±)-Aspamide D (**4**): colorless gum; [α] 25 <sup>D</sup> + 5.0 (*c* 0.05, MeOH); UV (MeOH) λmax (logε) 200 (6.07), 224 (6.06), 283 (5.33), 346 (5.60) nm; 1H and 13C NMR (DMSO-*d6*), see Table 2; positive HR-ESI-MS *m*/*z* 394.2122 [M + H]<sup>+</sup>, (calcd. for C23H28N3O3, 394.2125).

Aspamide F (**5**): brown powder; [α] 25 <sup>D</sup> + 40.0 (*c* 0.05, MeOH); ECD (5 mg/L, MeOH) λmax (Δε) 212 (65.33), 220 (62.98), 237 (−52.79) nm; UV (MeOH) λmax (logε) 206 (5.95), 224 (6.15), 270 (5.25), 278 (5.23) nm; 1H and 13C NMR (CDCl3), see Table 2; positive HR-ESI-MS *m*/*z* 336.1336 [M + H]+, (calcd. for C19H18N3O3, 336.1343).

Aspamide G (**6**): brown powder; [α] 25 <sup>D</sup> + 56.0 (*c* 0.05, MeOH); ECD (5 mg/L, MeOH) λmax (Δε) 212 (76.04), 221 (75.42), 237 (-62.18) nm; UV (MeOH) λmax (logε) 205 (5.92), 222 (5.88), 270 (5.16), 278 (5.13) nm; 1H and 13C NMR (CDCl3), see Table 2; positive HR-ESI-MS *m*/*z* 350.1498 [M + H]+, (calcd. for C20H20N3O3, 350.1499).
