Investigating Glycol-Split-Heparin-Derived Inhibitors of Heparanase: A Study of Synthetic Trisaccharides
<p>Schematic representation of roneparstat and necuparanib. The actual structure may retain the microheterogeneity of the original heparin and Low Molecular Weight Heparin.</p> "> Figure 2
<p>Dose-dependent inhibition of heparanase activity by SST001 (orange circles, IC<sub>50</sub>: 6 ng/mL); compound <b>2</b> (blue diamonds, IC<sub>50</sub>: 2 µg/mL) and compound <b>1</b> (green triangles, IC<sub>50</sub>: 30 µg/mL).</p> "> Figure 3
<p>Trisaccharide predicted conformation for compound <b>2</b> and compound <b>1</b> adjusting the backbone dihedral angles: τ, α/β, γ, δ/ε, ω/w for the former and τ, α/β, ω/w for the latter, in accord to the average values estimated using the MD simulation sampling procedure (material and methods). Possible extra-residue (GlcA) hydrogen bonds, contributing to <b>2</b> and <b>1</b> conformation stability, are underlined by dashed lines joining donor and acceptors atoms.</p> "> Figure 4
<p>(<b>left</b>) Selected experimental and simulated NOEs signals build up curves for compound <b>2</b> measured between protons H-1′′/H-5′ across the glycosidic linkage GlcNS6S-gsGlcA (empty circles, dotted line), and between H-1′/H-5′ inside the gsGlcA residue (empty squares, continuous line). Symbols and lines indicate experimental and simulated NOEs respectively; (<b>right</b>) Compound <b>2</b> and compound <b>1</b> predicted conformations, represented by cyan and orange tubes respectively are superposed by the GlcNS6S residue. Selected distances: H-1′′/H-4′, H-1′′/H-5′, and H-1′/H-5′ are expressed in Ǻ.</p> "> Scheme 1
<p>Conversion of <b>1</b> into <b>2</b>. Reagents and conditions: (i) NaIO<sub>4</sub>, H<sub>2</sub>O; (ii) NaBH<sub>4</sub>, 70%.</p> "> Scheme 2
<p>Synthesis of <b>1</b>. <span class="html-italic">Reagents and conditions</span>: (a) BF<sub>3</sub>:Et<sub>2</sub>O, 4 Å MS, CH<sub>2</sub>Cl<sub>2</sub>, −30 °C, 2 h, 77%; (b) 0.5 M MeONa, THF-MeOH, RT, 1 h; (c) PhCH(OMe)<sub>2</sub>, CSA, DMF, RT, 17 h, 56% (2 steps); (d) BnBr, NaH, DMF, RT, 2 h; (e) 70% aq. TFA, CH<sub>2</sub>Cl<sub>2</sub>, RT, 4 h; (f) (i) Dibromantin, TEMPO, CH<sub>3</sub>CN, RT, 2.5 h; (ii) CH<sub>3</sub>I, NaHCO<sub>3</sub>, DMF, RT, 16 h, 82% (4 steps); (g) TMSOTf, 4 Å MS, CH<sub>2</sub>Cl<sub>2</sub>, −20 °C, ~2 h, 58% (+ β-anomer 20%); (h) LiOH, H<sub>2</sub>O<sub>2</sub>, THF-MeOH, 16 h, 82%; (i) SO<sub>3</sub>:C<sub>5</sub>H<sub>5</sub>N, C<sub>5</sub>H<sub>5</sub>N, 55 °C, 2 h, 76%; (j) H<sub>2</sub>, 10% Pd/C, tBuOH-H<sub>2</sub>O, 60 h, 100%; (k) SO<sub>3</sub>:C<sub>5</sub>H<sub>5</sub>N, NaOH, RT, 16 h, 57%.</p> "> Scheme 3
<p>Synthesis of imidate <b>7</b>. <span class="html-italic">Reagents and conditions</span>: (a) CH<sub>3</sub>I, NaH, DMF, RT, 2 h, 100% (crude); (b) Ac<sub>2</sub>O-TFA, RT, 16 h, 100% (crude); (c) BnNH<sub>2</sub>, THF, RT, 3 h, 100% (crude); (d) Cl<sub>3</sub>CCN, CH<sub>2</sub>Cl<sub>2</sub>, Cs<sub>2</sub>CO<sub>3</sub>, RT, 3 h, 60% (4 steps).</p> ">
Abstract
:1. Introduction
2. Results and Discussion
2.1. Chemistry
2.2. Heparanase Inhibition Assay In Vitro
2.3. Conformation Characterization
3. Materials and Methods
3.1. General Information
3.2. Syntheses
3.3. MM/MD Simulation
4. Conclusions
Supplementary Materials
Acknowledgments
Author Contributions
Conflicts of Interest
References
- Xu, D.; Esko, J.D. Demystifying heparan sulfate-protein interactions. Ann. Rev. Biochem. 2014, 83, 129–157. [Google Scholar] [CrossRef] [PubMed]
- Ramani, V.C.; Purushothaman, A.; Stewart, M.D.; Thompson, C.A.; Vlodavsky, I.; Au, J.L.-S.; Sanderson, R.D. The heparanase/syndecan-1 axis in cancer: Mechanisms and therapies. FEBS J. 2013, 280, 2294–2306. [Google Scholar] [CrossRef] [PubMed]
- Vlodavsky, I.; Iozzo, R.V.; Sanderson, R.D. Heparanase: Multiple functions in inflammation, diabetes and atherosclerosis. Matrix Biol. 2013, 32, 220–222. [Google Scholar] [CrossRef] [PubMed]
- Casu, B.; Lindahl, U. Structure and biological interactions of heparin and heparan sulfate. Adv. Carbohydr. Chem. Biochem. 2001, 57, 159–208. [Google Scholar] [PubMed]
- Wilson, J.C.; Laloo, A.E.; Singh, S.; Ferro, V. 1H-NMR spectroscopic studies establish that heparanase is a retaining glycosidase. Biochem. Biophys. Res. Commun. 2014, 443, 185–188. [Google Scholar] [CrossRef] [PubMed]
- Pikas, D.S.; Li, J.P.; Vlodavsky, I.; Lindahl, U. Substrate specificity of heparanases from human hepatoma and platelets. J. Biol. Chem. 1998, 273, 18770–18777. [Google Scholar] [CrossRef] [PubMed]
- Okada, Y.; Yamada, S.; Toyoshima, M.; Dong, J.; Nakajima, M.; Sugahara, K. Structural recognition by recombinant human heparanase that plays critical roles in tumor metastasis. Hierarchical sulfate groups with different effects and the essential target disulfated trisaccharide sequence. J. Biol. Chem. 2002, 277, 42488–42495. [Google Scholar] [CrossRef] [PubMed]
- Wu, L.; Viola, C.M.; Brzozowski, A.M.; Davies, G.J. Structural characterization of human heparanase reveals insights into substrate recognition. Nat. Struct. Mol. Biol. 2015, 22, 1016–1022. [Google Scholar] [CrossRef] [PubMed]
- Lindahl, U.; Kjellen, L. Pathophysiology of heparan sulphate: Many diseases, few drugs. J. Intern. Med. 2013, 273, 555–571. [Google Scholar] [CrossRef] [PubMed]
- Rivara, S.; Milazzo, F.M.; Giannini, G. Heparanase: A rainbow pharmacological target associated to multiple pathologies including rare diseases. Future Med. Chem. 2016, 8, 647–680. [Google Scholar] [CrossRef] [PubMed]
- Alekseeva, A.; Mazzini, G.; Giannini, G.; Naggi, A. Structural features of heparanase-inhibiting non-anticoagulant heparin derivative roneparstat. Carbohydr. Polym. 2017, 156, 470–480. [Google Scholar] [CrossRef] [PubMed]
- Zhou, H.; Roy, S.; Cochran, E.; Zouaoui, R.; Chu, C.L.; Duffner, J.; Zhao, G.; Smith, S.; Galcheva-Gargova, Z.; Karlgren, J.; et al. M402, a novel heparan sulfate mimetic, targets multiple pathways implicated in tumor progression and metastasis. PLoS ONE 2011, 6, e21106,. [Google Scholar] [CrossRef] [PubMed]
- Vlodavsky, I.; Llan, N.; Naggi, A.; Casu, B. Heparanase: Structure, biological functions, and inhibition by heparin-derived mimetics of heparan sulfate. Curr. Pharm. Des. 2007, 13, 2057–2073. [Google Scholar] [CrossRef] [PubMed]
- Naggi, A.; Casu, B.; Perez, M.; Torri, G.; Cassinelli, G.; Penco, S.; Pisano, C.; Giannini, G.; Ishai-Michaeli, R.; Vlodavsky, I. Modulation of the heparanase-inhibiting of heparin through selective desulfation, graded acetylation, and glycol splitting. J. Biol. Chem. 2005, 280, 12103–12113. [Google Scholar] [CrossRef] [PubMed]
- Naggi, A. Glycol-splitting as a device for modulating inhibition of growth factors and heparanase inhibition by heparin and heparin derivative. In Chemistry and Biology of Heparin and Heparan Sulfate; Garg, H.G., Linhardt, R.J., Hales, C.A., Eds.; Elsevier: Amsterdam, The Netherland, 2005; pp. 461–481. [Google Scholar]
- Pala, D.; Rivara, S.; Mor, M.; Milazzo, F.M.; Roscilli, G.; Pavoni, E.; Giannini, G. Kinetic analysis and molecular modeling of the inhibition mechanism of roneparstat (SST0001) on human heparanase. Glycobiology 2016, 26, 640–654. [Google Scholar] [CrossRef] [PubMed]
- Petitou, M.; van Boeckel, C.A.A. A Synthetic Antithrombin III Binding Pentasaccharide Is Now a Drug! What Comes Next? Angew. Chem. Int. Ed. 2004, 43, 3118–3133. [Google Scholar] [CrossRef] [PubMed]
- Paulsen, H.; Stenzel, W. Building blocks of oligosaccharides. Synthesis of α-glycoside-linked 2-aminosugar oligosaccharides. Angew. Chem. 1975, 87, 547–548. [Google Scholar] [CrossRef]
- Oikawa, M.; Shintaku, T.; Sekljic, H.; Fukase, K.; Kusumoto, S. Synthesis of C-13-labeled biosynthetic precursor of lipid A and its analogue with shorter acyl chains. Bull. Chem. Soc. Jpn. 1999, 72, 1857–1867. [Google Scholar] [CrossRef]
- Grundler, G.; Schmidt, R.R. Glycosyl imidates, 13. Application of the trichloroacetimidate procedure to 2-azidoglucose and 2-azidogalactose derivatives. Liebigs Ann. Chem. 1984, 11, 1826–1847. [Google Scholar] [CrossRef]
- Takeda, N.; Ikeda-Matsumi, R.; Ebara-Nagahara, K.; Otaki-Nanjo, M.; Taniguchi-Morita, K.; Nanjo, M.; Tamura, J.I. Synthesis of heparan sulfate tetrasaccharide as a substrate for human heparanase. Carbohydr. Res. 2012, 353, 13–21. [Google Scholar] [CrossRef] [PubMed]
- Koshida, S.; Suda, Y.; Fukui, Y.; Ormsby, J.; Sobel, M.; Kusumoto, S. Synthesis and biological activity of oligomer-model compounds containing units of a key platelet-binding disaccharide of heparin. Tetrahedron Lett. 1999, 40, 5725–5728. [Google Scholar] [CrossRef]
- Hammond, E.; Li, C.P.; Ferro, V. Development of a colorimetric assay for heparanase activity suitable for kinetic analysis and inhibitor screening. Anal. Biochem. 2010, 396, 112–116. [Google Scholar] [CrossRef] [PubMed] [Green Version]
- Guerrini, M.; Elli, S.; Gaudesi, D.; Torri, G.; Casu, B.; Mourier, P.; Herman, F.; Boudier, C.; Lorenz, M.; Viskov, C. Effects on molecular conformation and anticoagulant activities of 1,6-anhydrosugars at the reducing terminal of antithrombin-binding octasaccharides isolated from low-molecular-weight heparin enoxaparin. J. Med. Chem. 2010, 53, 8030–8040. [Google Scholar] [CrossRef] [PubMed]
- Alekseeva, A.; Elli, S.; Cosentino, C.; Torri, G.; Naggi, A. Susceptibility of enoxaparin reducing end amino sugars to periodate oxidation. Carbohydr. Res. 2014, 400, 33–43. [Google Scholar] [CrossRef] [PubMed]
- Casu, B.; Guerrini, M.; Naggi, A.; Perez, M.; Torri, G.; Ribatti, D.; Carminati, P.; Giannini, G.; Penco, S.; Pisano, C.; et al. Short heparin sequences spaced by glycol-Split uronate residues are antagonists of fibroblast growth factor 2 and angiogenesis inhibitors. Biochemistry 2002, 41, 10519–10528. [Google Scholar] [CrossRef] [PubMed]
- Casu, B.; Guerrini, M.; Guglieri, S.; Naggi, A.; Perez, M.; Torri, G.; Cassinelli, G.; Ribatti, D.; Carminati, P.; Giannini, G.; et al. Undersulfated and glycol-split heparins endowed with antiangiogenic activity. J. Med. Chem. 2004, 47, 838–848. [Google Scholar] [CrossRef] [PubMed]
- Sample Availability: Not availabe.
© 2016 by the authors. Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC-BY) license ( http://creativecommons.org/licenses/by/4.0/).
Share and Cite
Ni, M.; Elli, S.; Naggi, A.; Guerrini, M.; Torri, G.; Petitou, M. Investigating Glycol-Split-Heparin-Derived Inhibitors of Heparanase: A Study of Synthetic Trisaccharides. Molecules 2016, 21, 1602. https://doi.org/10.3390/molecules21111602
Ni M, Elli S, Naggi A, Guerrini M, Torri G, Petitou M. Investigating Glycol-Split-Heparin-Derived Inhibitors of Heparanase: A Study of Synthetic Trisaccharides. Molecules. 2016; 21(11):1602. https://doi.org/10.3390/molecules21111602
Chicago/Turabian StyleNi, Minghong, Stefano Elli, Annamaria Naggi, Marco Guerrini, Giangiacomo Torri, and Maurice Petitou. 2016. "Investigating Glycol-Split-Heparin-Derived Inhibitors of Heparanase: A Study of Synthetic Trisaccharides" Molecules 21, no. 11: 1602. https://doi.org/10.3390/molecules21111602