Synthesis of biotinylated probes of artemisinin for affinity labeling

In this data article, we described the synthetic routes to four biotinylated probes (2, 3, 4, and 5) of artemisinin and the associated experimental procedures. We also provided the physical data for the synthesized compounds. These synthesized biotinylated probes of artemisinin are useful molecular tools for the affinity-labeling study of target receptor proteins of artemisinin in tropical pathogens such as Trypanosoma, Leishmania, and Schistosoma. The data provided herein are related to “Biotinylated probes of artemisinin with labeling affinity toward Trypanosoma brucei brucei target proteins”, by Konziase (Anal. Biochem. (2015)).


How data was acquired
Chemical reactions; normal phase column chromatography; NMR spectroscopy: JNM-GX-500 (JEOL), Lambda 500 (JEOL), Inova 600 (Varian); mass spectroscopy: JMS SX-102 (JEOL); IR spectroscopy: FT-IR-5300 (JASCO); polarimetry: DIP-370 (JASCO) Data format Analyzed, text, schemes Experimental factors N/A Experimental features Chemical reactions were performed under argon gas unless otherwise indicated; the diazirinecontaining probes were synthesized in brown opaque chemical flasks or transparent chemical flasks wrapped with aluminium foil due to photosensitivity. Data source location Osaka, Japan Data accessibility Data are available with this article

Value of the data
To reproduce all the experiments described in the research article ref [1]. To detect and isolate trypanosomal candidate target proteins of artemisinin.
To study the target receptors of artemisinin in Leishmania or Schistosoma.

General
1 H-NMR and 13 C-NMR spectra in CDCl 3 or CD 3 OD with TMS as the internal standard were recorded using a JNM-GX-500 or Lambda 500 (JEOL, Tokyo, Japan) NMR spectrometer operating at 500 MHz and 125 MHz, respectively. 2D NMR data in CDCl 3 were recorded using a Varian Inova 600 (Varian, Tokyo, Japan) NMR spectrometer operating at 600 MHz. Chemical shifts (δ) were reported in parts per million (ppm) and the multiplicities were designated as follows: s (singlet), d (doublet), t (triplet), m (multiplet), dd (doublet of doublets), ddd (doublet of doublet of doublets), brd (broad doublet), tlike (triplet like), dt (doublet of triplets). The coupling constants (J) were reported in Hz. Fast atom bombardment (FAB) and high-resolution fast atom bombardment (HR-FAB) mass spectra were recorded with a JMS SX-102 (JEOL, Tokyo, Japan) spectrometer in positive ion mode using magic bullet (5:1 dithiothreitol/dithioerythritol; Tokyo Kasei Kogyo) or m-nitrobenzyl alcohol as the matrix. Infrared (IR) spectra were recorded by a diffusion-reflection method on KBr powder using an FT-IR-5300 (JASCO, Tokyo, Japan) spectrometer. Shoulder bands in the IR spectra were designated by sh. Optical rotations were measured in a 0.5 dm length cell with a DIP-370 (JASCO, Tokyo, Japan) digital polarimeter. For column chromatography, silica gel (Fuji Sylisia BW-200 or Merck 60-230 mesh) and octadecyl silane ODS (Cosmosil 75C 18 OPN, Nacalai-Tesque) were used. Chemical reactions were performed under Ar gas unless otherwise indicated. TLC analyses were performed using normalphase pre-coated plates (Kiesel gel 60F 254 , Merck) and reversed-phase high-performance thin-layer chromatography (HPTLC) plates (RP-18 WF 254S , Merck). The spots on the thin-layer chromatograms were detected under UV light at 254 and 366 nm and visualized with either p-anisaldehyde/H 2 SO 4 (5 mL of AcOH, 25 mL of c-H 2 SO 4 , 425 mL of EtOH, and 25 mL of water) or phosphomolybdic acid (5 g in 100 mL of EtOH) spraying reagents and subsequent heating.
Preparation of probe 3 from 9: To a solution of 6 (5 mg, 0.007 mmol) in THF (0.2 mL) were added 2,4,6-trichlorobenzoyl chloride (1.1 μL, 1.7 mg, 0.9 mol equiv. to 6) and Et 3 N (2.14 μL, 1.56 mg, 2 mol equiv. to 6), and the entire mixture was stirred at room temperature overnight (ca. 15 h). Then, the previously prepared 9 (5.1 mg, 0.016 mmol) was added, and the mixture was stirred for 1 h at room temperature. Next, DMAP (0.47 mg, 0.5 mol equiv. to 6) was added, and an additional stirring was performed overnight at room temperature. The reaction mixture was directly evaporated under reduced pressure, affording a crude product (18.2 mg) that was applied to SiO 2 column chromatography (

Synthetic route to the biotinylated photoaffinity probe 4
We began the process with γ-butyrolactone (10) that underwent methanolysis in the first step, followed by protection of the primary alcohol with tert-butyldimethylsilyl chloride (TBSCl) in dichloromethane in the second step, affording 11, which was hydrolyzed under basic conditions to yield the tert-butyldimethylsilyl (TBS)-ether carboxylic acid 12. Condensation of 12 with 7 under EDCI Á HCl and DMAP in tetrahydrofuran (THF) led to 13, which was deprotected using a Dowex cation resin (50WX8, 100-200 mesh, H Cation Exchange Resin, Sigma-Aldrich) in MeOH, affording 14. Finally, condensation of 14 with 6 using EDCI Á HCl and DMAP in THF afforded probe 4 in a 28% yield (Scheme 3).
Preparation of 13 from 12: To a solution of 12 (19 mg, 0.087 mmol) in THF (1.2 mL) were added 7 (6.2 mg, 0.022 mmol), EDCI Á HCl (31.68 mg, 2 mol equiv. to 12), and DMAP (5.30 mg, 0.5 mol equiv. to 12). The mixture was stirred at room temperature for 2 h. Then, EDCI Á HCl (31.68 mg) and DMAP (5.30 mg) were added again, and an additional stirring was performed for 2 h. Following a reaction work up with brine, the mixture was extracted with EtOAc. The EtOAc layer was washed once with 5% HCl, once with saturated aqueous NaHCO 3 , once with brine, and then dried over MgSO4. Subsequent evaporation under reduced pressure produced 13 (18 mg) quantitatively (Scheme 3).
Compound Preparation of probe 4 from 14: To a solution of 6 (0.7 mg, 0.001 mmol) in THF-CH 3 CN (1:1, 90 μL) were added the previously prepared 14 (0.8 mg, 0.002 mmol), EDCI Á HCl (0.63 mg, 3 mol equiv. to 6), and DMAP (0.07 mg, 0.5 mol equiv. to 6), and then the mixture was stirred at room temperature for 3 h. Then, EDCI Á HCl (0.63 mg) and DMAP (0.07 mg) were added, and an additional stirring was performed at room temperature for 30 min, followed by another stirring performed at 40 1C for 2 h. Subsequently, the reaction mixture was evaporated under reduced pressure, affording a crude product (4.2 mg) that was applied to SiO 2 column chromatography (CHCl 3

Synthetic route to the biotinylated affinity probe 5
We started with d-biotin (15) that underwent a Curtius rearrangement in the first step, followed by condensation with tetraethylene glycol (16) in the second step to afford 17, which underwent a Michael addition to 18 and then hydrolyzed under basic conditions to afford the affinity labeling unit 19. Condensation of 19 with 7 using EDCI Á HCl and DMAP in THF-CH 3 CN¼1:1 afforded probe 5 in a 68% yield (Scheme 4).