Supplementary Materialsmolecules-24-01100-s001. DMSO-= 8.1 Hz, = 1.5 Hz, 1H), 4.09C4.14 (m, 1H), 3.70C3.80 (m, 2H), 3.44C3.60 (m, 2H), 3.20C3.38 (m, 2H), 2.72C2.89 (m, 6H), 2.55 (t, = 1.7 Hz, 1H), 2.46 (t, = 1.7 Hz, 1H), 2.25C2.38 (m, 4H), 1.82C1,92 (m, 2H), 1.60C1.78 (m, 4H), 1.36C1.50 (m, 4H), 1.20C1.35 (m, 4H), 1.13C1.18 (m, 1H), 1.04 (d, = 6.0 Hz, 1H), 0.92 (dd, =16.0 Hz, = 7.3 Hz, 3H), 0.7 (d, = 8.3 Hz, 3H). 13C-NMR (100 MHz, DMSOD6, 300 K) 174.9, 167.1, 160.0, 159.8, 157.3, 156.3, 140.1, 129.7, 121.6, 114.9, 112.5, 62.8, 62.6, 59.5, 57.8, 56.3, 55.9, 55.4, 53.8, 48.9, 56.4, 45.7, 44.7, 43.5, 41.0, 34.1, 33.7, 33.2, 28.2, 25.1, 23.4, 17.4, 14.2, 12.2, 11.5. MS (ESI) [M + H]+: 542.0. (TPI-2359-31): 1H-NMR (400 MHz, DMSO-= 6.4 Hz, 3H). 13C-NMR (100 MHz, DMSOD6, 300 K) 167.3, 159.9, 157,4, 62.8, 62.6, 59.4, 56.1, 55.3, 53.8, 46.2, 44.7, 42.1, 40.6, 34.1, 33.6, 29.3, 28.2, 25.1, 23.3, 19.6, 17.4, 15.3, 14.2, 12.2, 11.5. MS (ESI) [M Rabbit Polyclonal to Chk2 (phospho-Thr387) + H]+: 463.0. (TPI-2359-32): 1H-NMR (400 MHz, DMSO-= 6.8 Hz, 3H), 0.86 (d, = 6.6 Tonabersat (SB-220453) Hz, 6H), 0.72 (d, = 4.6 Hz, 3H). 13C-NMR (100 MHz, DMSOD6, 300 K) 167.3, 159.9, 157.3, 156.4, 62.8, 62.5, 59.4, 56.1, 56.1, 55.3, 53.8, 49.0, 46.2, 44.7, 42.5, 41.0, 35.4, 34.1, 33.7, 28.2, 27.7, 25.0, 24.0, 23.4, 17.3, 15.3, 14.2, 12.2, 11.5. MS (ESI) [M + H]+: 492.0. (TPI-2359-40): 1H-NMR (400 MHz, DMSO-=6.4 Hz, 3H), 0.91 (t, = 7.3 Hz, 3H), 0.87 (d, = 3.6 Hz, 6H), 0.82C0.87 (m, 3H), 0.75 (d, = 6.8 Hz, 3H). 13C-NMR (100 MHz, DMSOD6, 300 K) 174.4, 167.0, 160.3, 157.6, 62.9, 62.7, 59.5, 54.4, 54.0, 46.1, 45.2, 44.8, 41.3, 41.1, 35.3, 34.3, 29.4, 28.2, 27.6, 25.2, 24.4, 23.7, 23.5, 22.9, 22.8, 19.8, 19.4. MS (ESI) [M + H]+: 462.0. (TPI-2359-45): 1H-NMR (400 MHz, DMSO-= 7.3 Hz, 1H), 7.61 (d, = 7.6 Hz, 1H), 7.50C7.60 (m, 1H), 4.02C4.08 (m, 1H), 3.78C3.91 (m, 1H), 3.65C3.68 (m, 2H), 3.43C3.58 (m, 3H), 3.29C3.41 (m, 2H), 3.10C3.19 (m, 2H), 2.80C3.19 (m, 4H), 2.56C2.60 (m, 1H), 2.36C2.39 (m, 2H), 1.78C1.92 (m, 4H), 1.64C1.68 (m, 2H), 1.42C1.51 (m, 2H), 1.22C1.38 (m, 4H), 1.08C1.2 (m, 2H), 0.97 (d, = 6.5 Hz, 3H), 0,91 (dd, = 13.5 Hz, = 6.7 Hz, 6H), 0.83 (d, = 6.6 Hz, 3H), 0.749 (d, = 6.6 Hz, 3H). 13C-NMR (100 MHz, DMSOD6, 300 K) 167.3, 160.2, 157.2, 139.9, 133.7, 129.7, 126.1, 123.6, 63.1, 62.7, 59.4, 56.2, 54.4, 54.1, 46.7, 46.3, 43.1, 41.2, 34.4, 33.8, 32.4, 29.5, 28.3, 25.1, 23.9, 23.7, 19.8, 19.4, 17.6, 14.2, 12.1, 11.7. MS (ESI) [M + H]+: 592.0. (TPI-2359-47): 1H-NMR (400 MHz, DMSO-= 6.9 Hz, 6H), 0.82 (d, = 6.6 Hz, 3H), 0.73 (d, = 6.6 Hz, 3H). 13C-NMR (100 MHz, DMSOD6, Tonabersat (SB-220453) 300 K) 167.0, 160.3, 157.4, 63.1, 62.6, 59.3, 55.8, 54.1, 46.5, 46.1, 41.9, 41.2, 34.4, 33.9, 29.4, 28.8, 28.4, 25.2, 23.7, 23.6, 19.9, 19.8, 19.5, 17.5, 14.2, 14.1, 12.1, 11.8. MS (ESI) [M + H]+: 476.0 (TPI-2359-48): 1H-NMR (400 MHz, DMSO-= 6.6 Hz, 6H), 0.83 (d, = 6.6 Hz, 3H), 0.72 (d, = 6.6 Hz, 3H). 13C-NMR (100 MHz, DMSOD6, 300 K) 167.1, 160.3, 157.4, 63.0, 62.7, 59.5, 55.8, 54.0, 46.5, 46.0, 42.2, 41.2, 35.3, 34.4, 33.9, 29.4, 28.3, 27.7, 25.2, 24.5, 23.7, 22.9, 22.8, 19.8, 19.5, 17.5, 14.2, 12.1, 11.7, 11.4. MS (ESI) [M + H]+: 504.0. LCMS, 1H, and 13C-NMR data and spectra of all substances are included along with the structures of most 72 substances (TPI-2359) produced from the positional checking deconvolution from the collection TPI-1955. Balance data in mouse plasma of substance TPI 2359-47. Chemistry and Biology Experimental information (Supplementary Components). 3.1. P. falciparum Lifestyle and Antiplasmodial Activity Assay Dd2 (chloroquine-resistant) had been cultured utilizing a improved Trager and Jensen technique [42] in RPMI 1640 moderate with l-glutamine (Invitrogen, Carlsbad, CA, USA) and supplemented with 25 mM HEPES, pH 7.4, 26 mM NaHCO3, 2% dextrose, 15 mg/L hypoxanthine, 25 mg/L gentamycin, and 0.5% Albumax II in human A+ erythrocytes. Civilizations had been incubated at 37 C within a humidified environment of 5% CO2 and 95% surroundings. The compounds had been serially diluted in DMSO and had been put into the culture in a 1% parasitemia and 2% hematocrit in 96-well plates. The DMSO focus Tonabersat (SB-220453) hardly ever exceeded 0.125%. Pursuing 72-h incubation at 37 C, the viability from the parasite was driven utilizing a SYBR green I-based fluorescent assay which methods the DNA articles from the parasite [18,19,20]. Plates had been iced at ?80 C and thawed, accompanied by addition of 100 L of lysis buffer (20 mM Tris-HCl, 0.08% saponin, 5 mM EDTA, 0.8% Triton X-100, and 0.01% SYBR Green I) to each well. Pursuing incubation at night for 30 min at 37 C fluorescence emission in the.