Autopol 3

UV data were collected on a Hewlett Packard 8452A diode array spectrophotometer. Optical rotation data were determined on a Rudolph Research Analytical Autopol III automatic polarimeter. UV–CD spectra were measured on an AVIV circular dichroism spectrometer model 202-01. NMR data were obtained on a Varian VNMR spectrometer (500 MHz). Accurate mass data were collected on an Agilent 6538 HRESI QTOF MS coupled with an Agilent 1290 HPLC. LC-MS analyses were performed on a Shimadzu LC-MS 2020 system (ESI quadrupole) coupled to a photodiode array detector. The samples were separated using a Phenomenex Kintex column (2.6 µm C₁₈ column, 100 Å, 75×3.0 mm). The HPLC system utilized SCL-10A VP pumps and system controller with a Gemini 5 µm C₁₈ column (110 Å, 250×21.2 mm, flow rates of 1 to 10 mL/min). X-ray data were collected using a diffractometer with a Bruker APEX ccd area detector and graphite-monochromated Mo Kα radiation (λ = 0.71073 Å). All solvents were HPLC grade or better.

The optical rotations were recorded on a Rudolph Research Analytical Autopol III automatic polarimeter. NMR data were collected on a Bruker Avance II 600 MHz, high resolution 5-mm cryoprobe spectrometer operating at 600 MHz for ¹H and 150 MHz for ¹³C, using residual solvent signals (δ_H 2.50; δ_C 39.5 ppm DMSO-d₆, δ_H 2.50 ppm (CD₃)₂CO) as internal standards. The edited HSQC and HMBC experiments were optimized for ¹J_CH = 140 Hz ⁿJ_CH = 8 Hz, unless indicated otherwise (3 Hz HMBC). HRMS data were obtained using a Bruker Daltonics, Impact II QTOF with electrospray ionization (ESI). Chiral analysis was performed using an Agilent 6230 ESI-ToF and an Applied Biosystems 3200 QTRAP triple quad/linear trap. The effect on intracellular calcium was measured using a Molecular Devices Flexstation instrument. Circular Dichroism data were collected on a Chirascan™ Circular Dichroism Spectrometer (Applied Photophysics, Surrey, UK), with the Pro-Data Chirascan and Pro-Data viewer software 4.7.0.

Optical rotation was measured on a Rudolph Research Analytical AUTOPOL III automatic polarimeter. UV spectra were collected on a NanoDrop Spectrophotometer (Thermo Fisher Scientific, Inc., Waltham, MA, USA). NMR data was collected on a JEOL ECA-600 spectrometer (JEOL USA, Peabody, MA, USA) operating at 600 MHz for ¹H, and 150.9 for ¹³C. The edited gHSQC spectrum was optimized for 140 Hz and the gHMBC spectrum optimized for 8 Hz. Chemical shifts were referenced to solvent, e.g., CD₃OD, δ_H observed at 3.31 ppm and δ_C observed at 49.1 ppm. High-resolution mass spectrometry for 1 was performed on a JEOL AccuTOF-DART 4G (JEOL USA, Peabody, MA, USA) using the ESI source for ionization and detected in positive ion mode. High-resolution mass spectrometry for 2 was performed on a Thermo Fisher Orbitrap (Thermo Fisher Scientific, San Jose, CA, USA) using the ESI source for ionization and detected in positive ion mode. IR data was collected on a Perkin Elmer Spectrum 100 with Universal ATR (Perkin Elmer, Waltham, MA, USA).

The optical rotation was measured using an Autopol III (Rudolph Research Analytical, Hackettstown, NJ, USA) polarimeter with a 5-cm cell. The UV spectrum was recorded in MeOH on a S-2100 (Scinco, Seoul, Korea). The ECD spectrum was recorded using an Applied Photophysics Chirascan-Plus circular dichroism spectrometer (Applied Photophysics Ltd., Leatherhead, Surrey, UK). The IR spectrum was collected on a Varian Scimitar Series. NMR spectra were obtained using a Varian Inova NMR spectrometer (Varian, Inc., Palo Alto, CA, USA; 500 and 125 MHz for ¹H and ¹³C NMR, respectively), using the signals of the residual solvent as internal references (δ_H 3.31 and 4.78 ppm and δ_C 49.1 ppm for methanol-d₄). High resolution mass spectrum was obtained on a JMS-700 (JEOL Ltd., Tokyo, Japan) mass spectrometer. Low-resolution LC-MS data were analyzed using an Agilent Technologies 6120 quadrupole LC/MS system with a reversed-phase column (Phenomenex Luna C18(2) 100 Å, 50 mm × 4.6 mm, 5 μm) at a flow rate of 1.0 mL/min. Column chromatography separation was performed using C18 (40–63 μm, ZEO prep 90), eluting with a gradient of methanol and water. The fractions were purified using a reversed-phase HPLC Watchers 120 ODS-BP (250 mm × 10 mm, 5 μm) column, eluting with 80% CH₃CN in H₂O at flow rate of 2.5 mL/min.

Optical rotations were recorded on a Rudolph Research Analytical Autopol III automatic polarimeter. UV spectrophotometric data were acquired on a Shimadzu PharmaSpec UV–visible spectrophotometer. NMR data were collected on a JEOL ECA-600 spectrometer operating at 600.17 MHz for ¹H and 150.9 MHz for ¹³C. The edited-HSQC experiment was optimized for J_CH = 140 Hz and the HMBC spectrum was optimized for ^2/3J_CH = 8 Hz. ¹H NMR chemical shifts (referenced to residual CHCl₃ observed at δ 7.25 and residual CH₃OH observed at δ 3.30) were assigned using a combination of data from 2D DQF COSY and multiplicity-edited HSQC experiments. Similarly, ¹³C NMR chemical shifts (referenced to residual CHCl₃ observed at δ 77.0 and residual CH₃OH observed at δ 49.0) were assigned based on the multiplicity-edited HSQC experiments. The HRMS data were obtained using an Agilent 6210 LC-TOF mass spectrometer equipped with an APCI/ESI multimode ion source detector at the Mass Spectrometer Facility at the University of California, Riverside, California. Silica gel 60 (EMD Chemicals, Inc., Port Wentworth, GA, USA, 230–400 mesh) was used for column chromatography. All solvents used were of HPLC grade (Fisher Scientific, Waltham, MA, USA).

All reagents used in this study were HPLC grade or analytical grade solvent. Infrared spectra were acquired using a Thermo Nicolet iS10 IR spectrometer. CD spectra (ca. 0.5 mg/mL in methanol) were tested by a MOS-500 CD spectropolarimeter (Bio-Logic, France) in a 0.1 mm cuvette. Optical rotations were measured on an AUTOPOL III instrument (Rudolph Research Analytical, Hackettstown, USA). Nuclear magnetic resonance (NMR) was measured on a 700 MHz Avance III (Ascend) spectrometer by Bruker BioSpin GmbH, equipped each with a cryoplatform, at 25 °C. Spectra were recorded in 500 μL DMSO-d₆. Solvent signals were used as an internal standard (DMSO-d₆: δ_H 2.50, δ_C 39.5 ppm). High-resolution mass spectrometry was carried out on Agilent 6545 Quadrupole Time of Flight (Q-TOF) high-resolution mass spectrometer equipped with a reverse-phase C18 column (Agilent, Eclipse Plus, 50 × 2.1 mm, 1.8 μm), running in positive ionization mode with a resolution of 30,000. The flow rate was set at 0.3 mL/min with a mobile phase of H₂O/ACN each containing 0.1% of formic acid. The ACN percentage gradually increased from 5% to 95% in 12 min. The injection volume was 2 μL. Selected ions were fragmented using a collision-induced dissociation energy of 40 eV.