Click
here to close Hello! We notice that
you are using Internet Explorer, which is not supported by Echinobase
and may cause the site to display incorrectly. We suggest using a
current version of Chrome,
FireFox,
or Safari.
Mar Drugs
2025 May 30;236:. doi: 10.3390/md23060236.
Show Gene links
Show Anatomy links
Exploring the Inhibitory Effects of Fucosylated Chondroitin Sulfate (FCS) Oligosaccharide Isolated from Stichopus horrens and the Derivatives on P-Selectin.
Li C
,
Sun H
,
Gu X
,
Long W
,
Zhu G
,
Wu X
,
Wang Y
,
Li P
,
Sha L
,
Zhang J
,
Sun W
,
Gao N
,
Zuo Z
,
Zhao J
.
???displayArticle.abstract???
Unique fucosylated chondroitin sulfate (FCS) extracted from the sea cucumber Stichopus horrens was subjected to deacetylation and deaminative depolymerization to generate oligosaccharide fragments containing anTal-diol, which were further purified to obtain the trisaccharide ShFCS-3. Subsequently, the coupling of ShFCS-3 and 4-azidoaniline was achieved by reductive amination. After purification, the main product ShFCS-A1 and by-product ShFCS-A2 were obtained, which were identified as (N-(L-Fuc2S4S-α1,3-D-GlcA-β1,3-D-anTalA4S6S-1-)-4-azidoaniline) and (4S)-[2-(3-L-Fuc2S4S-α1)-D-GlcA-β1]-2,4,5-trihydroxy-5-sulfated-pent-2-enoic-acid) by 1D/2D NMR spectroscopy, respectively. ELISA experiments revealed that ShFCS-A1 exhibited P-selectin inhibition rates of 19.73% ± 9.60% at 1 μM, 96.28% ± 2.37% at 10 μM, and near-complete inhibition (99.92% ± 0.84%) at 100 μM. ShFCS-A2 demonstrated inhibition rates of 8.29% ± 3.00% at 1 μM, 74.02% ± 8.80% at 10 μM, and maximal inhibition approaching 100% at 100 μM. Cellular-level experiments revealed that ShFCS-A1 and ShFCS-A2 inhibited P-selectin binding to HL-60 cells by 92.72% ± 0.85% and 96.97% ± 1.16% at 100 μM, respectively. Molecular docking analysis indicated binding energies of -5.954 kcal/mol for ShFCS-A1 and -6.140 kcal/mol for ShFCS-A2 with P-selectin, confirming their potent inhibitory effects. These findings highlight the therapeutic potential of FCS oligosaccharides as pharmacophores and provide an important foundation for developing novel small-molecule P-selectin inhibitors.
Figure 1. Structures of CY-1503, PSI-697, GMI-1070, sLeX, TBC-1269, ShFCS-3, and 4-azidoaniline. ShFCS-3—trisaccharide derived from the sea cucumber Stichopus horrens. Fuc (F), fucosyl; GlcA (U), glucuronate; anTalA (T), 1-deoxy-2,5-anhydrate talosyl. P-selectin lectin/EGF domains were complexed with sLeX (PDB: 1G1R).
Figure 2. ShFCS extraction from Stichopus horrens.
Figure 3. HPGPC of ShFCS from Stichopus horrens (Shodex OHpak SB-804 HQ column). HPLC profiles of ShFCS, determined using a Shodex OHpak SB-804 HQ column (8 mm × 300 mm). Aliquots of 50 μL of 2 mg/mL sample were analyzed on an Agilent Technologies 1260 series apparatus equipped with a refractive index (RI) detector eluted with 0.1 M NaCl at a flow rate of 0.5 mL/min.
Figure 4. 1H NMR of ShFCS and deacetylated product of ShFCS.
Figure 5. HPGPC of dShFCS (Superdex peptide 10/300 GL). The HPLC profile of ShFCS-3 was analyzed using a Superdex peptide 10/300 GL column (10 mm × 300 mm). A sample of ShFCS-3 (50 μL, 2 mg/mL) was injected and eluted with 0.2 M NaCl at a flow rate of 0.4 mL/min.
Figure 6. The elution profile of ShFCS-3 (A), 1H NMR of ShFCS-3 (B), HPLC of ShFCS-3 (Superdex peptide 10/300 GL) and Structure of ShFCS-3 (C). he HPLC profile of ShFCS-3 was analyzed using a Superdex peptide 10/300 GL column (10 mm × 300 mm). A sample of ShFCS-3 (50 μL, 2 mg/mL) was injected and eluted with 0.2 M NaCl at a flow rate of 0.4 mL/min.
Figure 7. Synthetic strategy for ShFCS-3 derivatives.
Figure 8. The synthesis route of ShFCS-3 derivatives. ShFCS-A1: N-(L-Fuc2S4S-α1,3-D-GlcA-β1,3-D-anTalA4S6S-1-)-4-azidoaniline; ShFCS-A2: (4S)-[2-(3-L-Fuc2S4S-α1)-D-GlcA-β1]-2,4,5-trihydrox-y-5-sulfated-pent-2-enoic acid; PenA (A), 2,4,5-trihydroxy-5-sulfated-pent-2-enoic acid.
Figure 9. Elution curve of ShFCS-A1 and ShFCS-A2 on Bio-Gel P-2.
Figure 10. 1H (A); 13C (B); 1H-1H COSY (black), TOCSY (red), and ROESY (blue) (C); 1H-13C HSQC (D); and 1H-13C HMBC (E) of ShFCS-A1.
Figure 11. 1H (A); 13C (B); 1H-1H COSY (black), TOCSY (red), and ROESY (blue) (C); 1H-13C HSQC (D); and 1H-13C HMBC (E) of ShFCS-A2.
Figure 12. Possible process for the formation of the by-product ShFCS-A2.
Figure 13. Inhibitory effect of compounds on P-selectin binding to PSGL-1 (n = 3).
Figure 14. Inhibitory effects of compounds on the binding of P-selectin to HL-60 cells (n = 3).
Figure 15. Molecular docking results of sLeX, ShFCS-3, ShFCS-A1, and ShFCS-A2.
