Smart tools and orthogonal click-like reactions onto small unilamellar vesicles: Additional molecular data

We present here the synthetic routes and the experimental data (NMR and MS spectra) for model reactions for copper-free Huisgen 1,4-cycloaddition, Staudinger ligation and for addition of a dithiol on a dibromomaleimide ring. Starting materials were synthesized from the commercially available 4-chlorophenethylamine, previously described 2-(cyclooct-2-yn-1-yloxy)acetic acid, 1-fluorocyclooct-2-ynecarboxylic acid, commercial 2-(diphenylphosphino)terephthalic acid 1-methyl 4-pentafluorophenyl diester and dibromomaleimide. In all cases, the expected compounds were obtained with good yield (50% to quantitative). A novel synthesis of the lipid anchor DOGP3NH2 is also described. These data were used as basis for the study reported in the article “Smart Tools and Orthogonal Click-like Reactions onto Small Unilamellar Vesicles” in Chemistry and Physics of Lipids [1].


Subject area Chemistry
More specific subject area Bioconjugation Type of data Experimental synthesis protocols, analysis description, NMR and MS spectra How data was acquired 1 H NMR spectra at either 300 MHz, 400 MHz or 500 MHz and 13 C NMR spectra at either 75 MHz, 100 MHz or 133 MHz recorded on Brucker spectrometers either 300, 400 or 500 respectively with residual undeuterated solvent as internal reference. High-resolution mass spectra (HRMS) obtained using an Agilent Q-TOF (time of flight) 6520 and low-resolution mass spectra (LRMS) using an Agilent MSD 1200 SL (ESI/APCI). Analytical RP-HPLC-MS performed using a C18 column (30 mm Â 1 mm; 1.9 μm) using the following parameters: (1) the eluent system A (0.05% TFA in H 2 O) and B (0.05% TFA in acetonitrile); (2) the linear gradient t ¼0 min with 98% A, t ¼5 min with 5% A, t ¼6 min with 5% A, t¼ 7 min with 98% A, and t ¼9 min with 98% A; (3) flow rate of 0.3 mL min -1 ; (4) column temperature 50 1C; (5) ratio of products determined by integration of spectra recorded at 210 or 254 nm; and (6) ionization mode ESI. Data format Analyzed data Experimental factors Starting compounds were either purchased or synthesized using already published synthetic protocols Experimental features Compounds were synthesized and their structure was identified by NMR and confirmed by mass spectrometry Data source location Illkirch, France Data accessibility Data are provided in the paper

Value of the data
The data presented prove the efficiency of bioconjugation reactions and allow reproducibility of the syntheses.
The overall work provides further tools for surface modification of liposomes. The article describes a novel, easier synthesis of the lipid anchor DOGP 3 NH 2 .

Data
All the described 1 H NMR and 13 C NMR spectra, as well as MS spectra, are available as annexes to this article. 1 H NMR spectra at either 300 MHz, 400 MHz or 500 MHz and 13 C NMR spectra at either 75 MHz, 100 MHz or 133 MHz were recorded on Brucker spectrometers either 300, 400 or 500 respectively with residual undeuterated solvent as internal reference. All chemical shift values (δ), coupling constants (J) and the multiplicity (s ¼singlet, d ¼doublet, t¼triplet, m¼ multiplet, br¼ broad) are quoted in ppm and in Hz, respectively. High-resolution mass spectra (HRMS) were obtained using an Agilent Q-TOF (time of flight) 6520 and low-resolution mass spectra (LRMS) using an Agilent MSD 1200 SL (ESI/APCI).

Experimental design, materials and methods
Reagent grade solvents were used without further purification. Polymer supported triphenylphosphine and anhydrous CH 2 Cl 2 were purchased from Sigma-Aldrich. The PyBOP was purchased from Novabiochem, the DIEA was from Alfa Aesar and both were used without further purification. Column chromatography was carried out on silica gel 60 (Merck, 70À 230 mesh). Analytical RP-HPLC-MS was performed using a C18 column (30 mm Â 1 mm; 1.9 μm) using the following parameters: (1) the eluent system A (0.05% TFA in H 2 O) and B (0.05% TFA in acetonitrile); (2) the linear gradient t¼ 0 min with 98% A, t¼5 min with 5% A, t¼ 6 min with 5% A, t¼7 min with 98% A, and t¼9 min with 98% A; (3) flow rate of 0.3 mL min À 1 ; (4) column temperature 50 1C; (5) ratio of products determined by integration of spectra recorded at 210 or 254 nm; and (6) ionization mode ESI. TLC spots were detected by UV irradiation at 254 nm or with KMnO 4 stain.

Synthesis of the lipid anchors
See Scheme 1.