Isolera 1 system

An aliquot of the EtOAc fraction (7.9 g) was divided into 500 mg samples and each was subjected to flash MPLC (Isolera 1 System, Biotage, Sweden) under the following conditions: cartridge, silica HP 50 g; gradient, EtOAc (A) in n-hexane, 0% (A) for 198 mL to equilibrate, 0% (A) for 264 mL, 0% to 60% (A) for 1386 mL, 60% (A) to 100% (A) for 198 mL, and staying at 100% (A) for 66 mL; flow rate, 50 mL/min; detection, 270 nm; fraction volume, 22 mL. Twelve sub-fractions were collected by combining eluates corresponding to each major peak of the generated chromatogram. Each sub-fraction was concentrated using the rotary evaporator, transferred into pre-weighed capped vials and dried at 45°C in a Centrifan PE-T evaporator (kd Scientific, Holliston, MA). HPLC analysis was performed on each sub-fraction and compared with the total fingerprint to identify the major constituent(s) in each sub-fraction.

All peptides for on-bead and solution phase reactions were synthesized using standard Fmoc solid phase synthesis techniques on Rink amide or 2-chlorotrityl-functionalized polystyrene resins, respectively. Sequences were purified using a combination of normal-phase flash chromatography on silica and reverse phase chromatography on a Biotage Isolera 1 system. Explicit details for these procedures, MS/MS sequencing protocols, full characterization for peptides 1 and 12, and ¹H NMR and HRMS data for all other solution phase peptides are summarized in sections II, III, and VI of the Supporting Information. Characterization data for peptides 1 and 12 are shown below: