Lc it tof ms

Unless otherwise specified, reactions were performed under an inert atmosphere of argon and monitored by thin-layer chromatography (TLC) and/or LCMS. All reagents were purchased from commercial suppliers and used as provided. Synthetic intermediates were purified using CombiFlash chromatography system on 230–400 mesh silica gel. ¹H and ¹³C NMR spectra were obtained using Bruker DPX-400 or AVANCE-400 spectrometer at 400 and 100 MHz, respectively. NMR chemical shifts were described in δ (ppm) using residual solvent peaks as standard (chloroform-d, 7.26 ppm (¹H), 77.16 ppm (¹³C); methanol-d₄, 3.31 ppm (¹H), 49.00 ppm (¹³C); DMSO-d₆, 2.50 ppm (¹H), 39.52 ppm (¹³C); acetone-d₆, 2.05 ppm (¹H), 29.84 ppm (¹³C)). Data were reported in a format as follows: chemical shift, multiplicity (s = singlet, d = doublet, dd = doublet of doublet, t = triplet, q = quartet, br = broad, m = multiplet, abq = ab quartet), number of protons, and coupling constants. High resolution mass spectral data were measured in-house using a Shimadzu IT-TOF LC/MS for all final compounds. All compounds submitted for biological testing were confirmed to be ≥95% pure by analytical HPLC. Synthetic methods, spectral data, and HRMS for novel compounds are described in detail below.

Detailed methods are provided in Supporting Information, including characterization and purity. Unless otherwise specified, reactions were performed under an inert atmosphere of argon and monitored by thin-layer chromatography (TLC) and/or LCMS. All reagents and solvents were purchased from commercial suppliers (Sigma-Aldrich, Fisher Scientific, Ambeed, Combi-Blocks, Enamine) and used as provided. Synthetic intermediates were purified using a CombiFlash chromatography system on 230–400 mesh silica gel or Shimadzu prep-HPLC system. ¹H and ¹³C NMR spectra were obtained using Bruker DPX-400 or AVANCE-400 spectrometer at 400 and 100 MHz, respectively. NMR chemical shifts were described in δ (ppm) using residual solvent peaks as standard. High resolution mass spectral data were measured in-house using a Shimadzu IT-TOF LC/MS for all final compounds. Optical rotations were measured with a Perkin-Elmer 241 polarimeter operating on the mercury lamp line (546 nm), using a 100 mm pathlength cell. All compounds submitted for biochemical and biological testing were confirmed to be ≥ 95% pure by analytical HPLC.

3-chloro-6-methoxybenzo[b]thiophene-2-carbonyl chloride was purchased from Frontier Scientific Services, Inc. All chemicals and solvents were purchased from Sigma Aldrich, Fisher Scientific, Matrix Scientific or Oakwood Chemical and were used without further purification. 2 was synthesized using the protocols reported by Lai and colleagues.^{20 (link)} Synthetic intermediates were purified using Biotage flash chromatography system on 230–400 mesh silica gel. ¹H and ¹³C NMR spectra were obtained using Bruker DPX-400 or AVANCE-400 spectrometer at 400 and 100 MHz, respectively. NMR chemical shifts were described in δ (ppm) using residual solvent peaks as standard (CDCl₃, 7.26 ppm (¹H), 77.16 ppm (¹³C); CD₃OD, 3.31 ppm (¹H), 49.00 ppm (¹³C); DMSO-d₆, 2.50 ppm (¹H), 39.52 ppm (¹³C); Acetone-d₆, 2.05 ppm (¹H), 29.84 ppm (¹³C)). Data were reported in a format as follows: chemical shift, multiplicity (s = singlet, d = doublet, dd = doublet of doublet, t = triplet, q = quartet, br = broad, m = multiplet, abq = ab quartet), number of protons, and coupling constants. High resolution mass spectral data were measured in-house using a Shimadzu IT-TOF LC/MS for all final compounds. All compounds submitted for biological testing were confirmed to be ≥95% pure by analytical HPLC. Synthetic methods, spectral data, and HRMS for novel compounds are described in detail below.