Article Figures & Data
Figures
- Fig. 1.
Target polyphenols with two heads and one tail.
- Scheme 1.
Synthesis of the N-acyl serinol derivatives 2-6. Reagent and conditions: (i) MeOH:THF (2:1), Triethylamine, −20°C to room temperature (rt).
- Scheme 2.
Synthesis of 13-17. Reagent and conditions: (i) a) MOMCl, b) HOLi/H2O (ii) HATU, DMAP (iii) aqueous HCl (37%).
- Scheme 3.
Synthesis of 20. Reagent and conditions: (i) HATU, DMAP (ii) H2, Pd/C.
- Scheme 4.
Synthesis of 24. Reagent and conditions: (i) HATU, DMAP (ii) H2, Pd/C.
- Fig. 2.
Glycolytic activity in SW-620 colon cancer cells. Comparison of the glycolytic function in SW-620–nontreated cells versus 16-treated cells (0.83 and 1.66 µM) and 14-treated cells (0.83 and 16.66 µM). Glycolysis (difference from ECAR value in the presence of glucose and ECAR value in starved cells), glycolytic reserve (difference between ECAR after ATPase inhibition and ECAR after glucose injection), and glycolytic capacity (sum of glycolysis rate and glycolytic reserve) are shown. Representative assay of two experiments. Each experiment contains six replicates per treatment. Asterisk indicates statistical differences in treated cells with respect to the control (nontreated cells) *P < 0.05.
- Fig. 3.
Mitochondrial respiration in SW-620 colon cancer cells. (A) Mitochondrial respiration analysis by flux analysis of the OCR in SW-620–nontreated cells versus 16-treated cells (0.83 and 1.66 µM) and 14-treated cells (0.83 and 16.66 µM). (B) Mitochondrial respiration analysis by flux analysis of the OCR in SW-620–nontreated cells versus 16-treated cells (1.66 and 3.2 µM) and 14-treated cells (1.66 and 60 µM). Representative assays of two experiments. Each experiment contains six replicates per treatment. Asterisk indicates statistical differences in treated cells with respect to the control (nontreated cells) *P < 0.05.
- Fig. 4.
Compound 16 leads to AMPK activation. (A) Western blot analysis of P-(Thr172) AMPK and total AMPK. SW-620 cells were treated for 48 hours with compound 16 (d1: 0.83 µM; d2: 1.66 µM; and d3: 2.5 µM) and 14 (d1: 1.66 µM and d2: 29.5 µM). Representative experiment. (B) Western blot quantification of ratio P-(Thr172)AMPK/total AMPK. (C) Measurement of the intracellular ATP content. Luminiscence units were registered for compound 16-treated cells (0.83 and 1.66 µM) and compared with control nontreated cells. Asterisk indicates statistical differences in treated cells with respect to the control (nontreated cells) *P < 0.05.
- Fig. 5.
Caspase 3/7 activity. Activation of caspase 3 and 7 after 48-hour treatment with compounds 14 (1.66 µM) and 16 (1.66 µM). SW-620 cells treated with staurosporine (1.5 µM) were used as the positive control. Data represent means ± S.E.M. of two independent experiments, each performed in triplicate. Asterisks indicate statistical differences in treated cells with respect to the control (nontreated cells) *P < 0.05.
Tables
- TABLE 1
Inhibitory effects of test compounds on SW-620 human colon cancer cell viability
Data are the mean ± S.E.M. of at least three independent experiments, each performed in triplicate. Data (>100); not significant activity found at 150 µM concentration, after 48-hour treatment.
Compound IC50 (µM)a GI50 (µM)b TGI (µM)c LC50 (µM)d 13 >100 >100 >100 >100 14 29.5 ± 10.18 10.5 ± 1.55 56.25 ± 7.46 75.5 ± 5.42 15 2 ± 0 1.58 ± 0.08 10.33 ± 1.45 19.33 ± 2.85 16 0.83 ± 0.083 0.57 ± 0.067 14.47 ± 12.77 75 ± 25 17 2.67 ± 0.67 1.83 ± 0.083 >100 >100 20 43.33 ± 4.41 24.33 ± 8.29 >100 >100 24 5.17 ± 1.17 2.33 ± 0.33 36.67 ± 4.41 63.33 ± 8.33 SF-6 >100 >100 >100 >100 4,4′-Di-O-MEA 5.97 ± 1.09 1.58 ± 0.18 143.3 ± 23.66 >100 4,4′-Di-O-MEA, 4,4′-di-O-methylellagic acid.
↵a Effective concentration, or compound concentration required for 50% inhibition of cell proliferation, after 48-hour treatment.
↵b Compound concentration required for 50% cell growth inhibition, after 48-hour treatment.
↵c Compound concentration required for total cell growth inhibition, after 48-hour treatment.
↵d Compound concentration required for 50% cell death, after 48-hour treatment.
- TABLE 2
Sensitivity of SW-480 and SW-620 human colon cancer cells to compound 16
Data are the mean ± S.E.M. of at least three independent experiments, each performed in triplicate.
Colorectal Cancer Cell Line IC50 (µM)a GI50 (µM)b TGI (µM)c LC50 (µM)d SW-480 4.03 ± 0.183 2.67 ± 0.27 24.31 ± 15.32 84 ± 17 SW-620 0.83 ± 0.083 0.57 ± 0.067 14.47 ± 12.77 75 ± 25 ↵a Effective concentration, or compound concentration required for 50% inhibition of cell proliferation, after 48-hour treatment.
↵b Compound concentration required for 50% cell growth inhibition, after 48-hour treatment.
↵c Compound concentration required for total cell growth inhibition, after 48-hour treatment.
↵d Compound concentration required for 50% cell death, after 48-hour treatment.
Data Supplement
- Supplemental Data -
1H and 13C NMR spectra.
- Supplemental Data -
