400 mhz instrument

Commercially available starting materials, reagents, and solvents were used as received. Unless otherwise indicated, all reactions were conducted in oven-dried glassware. In general, anhydrous reactions were performed under nitrogen. Reactions were monitored by thin-layer chromatography (TLC) carried out on pre-coated glass plates of silica gel (0.25 mm) 60 F₂₅₄ from EMD Chemicals Inc. Visualization was accomplished with ultraviolet light (UV 254 nm), or by shaking the TLC plate in a sealed jar containing silica gel and iodine. Flash column chromatography was performed using 230–400 mesh silica gel purchased from Silicycle. All work-up and purification procedures were carried out with reagent grade solvents in the air. Yields refer to isolate yield unless otherwise stated. Melting points were determined on a MEL-TEMP 3.0 apparatus. ¹H NMR and ¹³C NMR spectra were recorded on Varian 400 MHz instrument. Chemical shifts are reported in parts per million (ppm) and are calibrated using residual undeuterated solvent as an internal reference (CDCl₃: δ 7.26 ppm; CD₃OD: δ 3.31 ppm; DMSO: δ 2.50 ppm). Data are reported as follows: chemical shift, multiplicity, coupling constants (Hz), and integration. High resolution positive ion mass (HRMS) analyses were conducted on a Bruker MaXis 4G Q-TOF mass spectrometer with electrospray ionization source.

¹H (400 MHz) and ¹³C (101 MHz) NMR spectra were acquired on a Varian 400 MHz instrument at the University of Las Vegas, Nevada. Chemical shifts are reported in ppm (δ) and are referenced to DMSO-d₆ (2.50 ppm for ¹H and 39.5 ppm for ¹³C) or CDCl₃ (7.27 ppm for ¹H and 77.0 ppm for ¹³C). Coupling constants J_HH are in hertz and are reported as follows: chemical shift, multiplicity (br = broad, s = singlet, d = doublet, t = triplet, q = quartet, m = multiplet, dd = doublet of doublets, dt = doublet of triplets), coupling constant, and integration. Melting points were acquired using a Stuart SMP10 Digital Melting Point Apparatus and are uncorrected. Infrared spectra (IR) were recorded with a Shimadzu IRAffinity-1 FT-IR spectrophotometer. UV/Vis measurements employed a Beckman Coulter DU-800 spectrophotometer. Fluorescence measurements used a Horiba Scientific FluoroMax-4 spectrofluorometer. High-resolution mass spectra were obtained at the Mass Spectrometry Facility at the University of California, Irvine. Thin layer chromatography (TLC) used aluminum-backed silica plates (0.20 mm, 60 F-254). Column chromatography and silica plugs were performed using Silicycle Siliaflash P60 silica. Plates were visualized by UV light. Commercial reagents were used as received unless otherwise noted. Yields are reported based on isolated material.

¹H NMR was taken on a Varian 400 MHz instrument and recorded at ambient temperature. DBI-1 was prepared in deuterated-DMSO at a concentration of 10 mg/mL. NMR was calibrated to the solvent peak at 2.5 ppm.

All solvents were reagent grade or HPLC grade, and all starting materials were obtained from commercial sources and used without further purification. A Thermo Finnigan LCQ Deca instrument with a Thermo Surveyor LCMS system at variable wavelengths of 254 and 214 nm was used to monitor the reactions. The purity of the final compounds was assessed using an Agilent 1100 HPLC instrument (Phenomenex kinetex 150 × 4.6 mm column, 5 μm; 254 nm wavelength) and final compounds purities were >95%. The HPLC mobile phase consisted of a water–methanol gradient (30–100% methanol) buffered with 0.1% formic acid, the running time of which was 15 min and the flow rate of which was 0.8 mL/min. ¹H NMR spectra were recorded at 400 MHz, and ¹³C spectra were recorded at 100 MHz. Both were completed on a Varian 400 MHz instrument (model no. 4001S41ASP). All compounds were purified using silica gel (0.035–0.070 mm, 60 Å) flash chromatography on a Teledyne ISCO CombiFlash systemunless otherwise noted. Microwave-assisted reactions were completed in sealed vessels using a Biotage Initiator microwave synthesizer.

All solvents and reagents were commercially available and used without further purification unless otherwise stated. 7-Methylumbeliferone and 3-phenylphenol were obtained from Sigma-Aldrich; 4-Phenoxypyridine was purchased from TCI America and used directly as a precursor for radiolabeling. NMR spectra were recorded with a Varian 400 MHz instrument at room temperature with tetramethylsilane (TMS) as an internal standard. Mass spectra were performed on a Micromass LCT Time-of-Flight mass spectrometer or an Agilent Q-TOF HPLC-MS employing the electrospray ionization (ESI) method. High performance liquid chromatography (HPLC) was performed using a Shimadzu LC-2010A HT system equipped with a Bioscan B-FC-1000 radiation detector.