*3.2. Fungal Material, Fermentation, and Isolation of Secondary Metabolites*

Fungal Material, Fermentation, and Isolation of **1**–**3** from *Aspergillus unguis* IV17-109

*A. unguis* IV17-109 (GenBank accession number OL700797) was isolated from a deepsea shrimp sample as previously described [10]. The EtOAc extract was fractionated into 10 fractions (F1–F10), as described earlier [10]. The F7 fraction was purified by a semipreparative reversed-phase HPLC (YMC-Pack-ODS-A, 250 × 10 mm i.d, 5 μm, flow rate 2.0 mL/min, 60% MeOH/H2O, RI detector) to obtain **3** (2.0 mg, *tR* = 32 min). The F8 fraction was subjected to a semi-preparative reversed-phase HPLC (YMC-Pack-ODS-A, 250 × 10 mm i.d, 5 μm, flow rate 2.0 mL/min, RI detector) using an isocratic elution with 70% MeOH/H2O to yield a subfraction F8-1, and the subfraction was further purified by a semi-preparative HPLC (YMC-Pack-ODS-A, 250 × 10 mm i.d, 5 μm, flow rate 2.0 mL/min, RI detector) using an isocratic elution with 50% MeCN/H2O to obtain **2** (1.0 mg, *tR* = 15 min). Finally, the F9 fraction was purified by a semi-preparative reversed-phase HPLC (YMC-Pack-ODS-A, 250 × 10 mm i.d, 5 μm, flow rate 2.0 mL/min, RI detector) using an isocratic elution with 83% MeOH/H2O to yield **1** (3.0 mg, *tR* = 42 min).

Variotin B (**1**): pale-yellow needles; IR νmax 3286, 2918, 1724, 1671, 1352, 1261, 989 cm<sup>−</sup>1; UV(MeOH) λmax (log ε) 283 (2.51), 229 (2.60) nm; HRESIMS *m*/*z* 336.1938 [M+Na]+ (calcd for C20H27NO2Na, 336.1939), 1H NMR (CD3OD, 600 MHz) and 13C NMR (CD3OD, 150 MHz) see Table 1.

Coniosulfide E (**2**): colorless solid, [α] 20 <sup>D</sup> − 100 (c 0.3, MeOH); IR νmax 3303, 2929, 1653, 1547, 1374, 1038 cm<sup>−</sup>1, HRESIMS *m*/*z* 451.2607 [M+Na]+ (calcd for C22H40N2O4SNa, 451.2606), 1H NMR (CD3OD, 600 MHz) and 13C NMR (CD3OD, 150 MHz) see Table 1.
