Synthesis and evaluation of galacto-noeurostegine and its 2- deoxy analogue as glycosidase inhibitors

Transcription

1 Electronic Supplementary Material (ESI) for Organic & Biomolecular Chemistry. This journal is The Royal Society of Chemistry 2015 Synthesis and evaluation of galacto-noeurostegine and its 2- deoxy analogue as glycosidase inhibitors Stéphane Salamone, a, Lise L. Clement, a Agnete H. Viuff, a Ole J. Andersen, a,b Frank Jensen a and Henrik H. Jensen*,a a Department of Chemistry, Aarhus University, Langelandsgade 140, DK-8000, Aarhus C, Denmark. hhj@chem.au.dk; Tel: b Center for Insoluble Protein Structures (inspin) and the Interdisciplinary Nanoscience Center (inano), Aarhus University, Langelandsgade 140, DK-8000, Aarhus C, Denmark. Present address: Oxeltis, Cap Gamma 1682, rue de la Valsière, CS 27384, Montpellier, France. Content Quantum mechanical calculation setups p 2 Michaelis-Menten and Hanes plots for compounds 5 and 6 p 4 1 H and 13 C NMR spectra for all new compounds p 8 1

2 Quantum mechanical calculation setups Noeurostegine, benzyl protected analogue: Conformational search: Open and closed forms of the molecule were run through a conformational search using the MCMM method implemented in the Schrödinger Suite. The search was performed using 10,000 steps in the implicit CHCl 3 model with no cutoff. Searches were performed using both the OPLS-2005 and MMFF force fields. Unique conformations (defined by a 0.5 Å RMSD cutoff using all heavy atoms) within 10 kj/mol of the lowest energy structure were kept for QM geometry optimisations. Geometry optimisation: The conformations obtained from the conformational search were DFT optimised using the pcseg 1 basis set and the wb97xd functional. The IEFPCM method was used to model the presence of CHCl 3. Conformations within 10 kj/mol of the lowest energy structure were kept for MP2 single point calculations. MP2 single point calculations: MP2 calculations were performed using the aug cc pvdz basis set and the wb97xd functional. The IEFPCM method was used to model the presence of CHCl 3. Noeurostegine: Conformational search: Charged and neutral configurations of the open and closed forms of the molecule were run through a conformational search using the MCMM method implemented in the Schrödinger Suite. The search was performed using 10,000 steps in the implicit water model with no cutoff. Searches were performed using both the OPLS-2005 and MMFF force fields. Unique conformations (defined by a 0.2 Å RMSD cutoff using all heavy atoms) within 35 kj/mol of the lowest energy structure were kept for QM geometry optimisations. Geometry optimisation: The conformations obtained from the conformational search were DFT optimised using the pcseg 1 basis set and the wb97xd functional. The IEFPCM method was used to model the presence of water. All conformations were kept for MP2 single point calculations. MP2 single point calculations: MP2 calculations were performed using the aug cc pvtz basis set and the wb97xd functional. The IEFPCM method was used to model the presence of water. Galacto-noeurostegine, benzyl protected analogue (28): Conformational search: Open and closed forms of the molecule were run through a conformational search using the MCMM method implemented in the Schrödinger Suite. The search was performed using 10,000 steps in the implicit CHCl 3 model with no cutoff. Searches were performed using both the OPLS-2005 and MMFF force fields. Unique conformations (defined by a 0.5 Å RMSD cutoff using all heavy atoms) within 35 kj/mol of the lowest energy structure were kept for QM geometry optimisations. Geometry optimisation: The conformations obtained from the conformational search were DFT optimised using the pcseg 1 basis set and the wb97xd functional. The IEFPCM method was used to model the presence of CHCl 3. Conformations within 20 kj/mol of the lowest energy structure were kept for MP2 single point calculations. MP2 single point calculations: MP2 calculations were performed using the aug cc pvdz basis set and the wb97xd functional. The IEFPCM method was used to model the presence of CHCl 3. 2

3 Galacto-noeurostegine (5): Conformational search: Charged and neutral configurations of the open and closed forms of the molecule were run through a conformational search using the MCMM method implemented in the Schrödinger Suite. The search was performed using 10,000 steps in the implicit water model with no cutoff. Searches were performed using both the OPLS-2005 and MMFF force fields. Unique conformations (defined by a 0.2 Å RMSD cutoff using all heavy atoms) within 50 kj/mol of the lowest energy structure were kept for QM geometry optimisations. Geometry optimisation: The conformations obtained from the conformational search were DFT optimised using the pcseg 1 basis set and the wb97xd functional. The IEFPCM Method was used to model the presence of water. Conformations within 20 kj/mol of the lowest energy structure were kept for MP2 single point calculations. MP2 single point calculations: MP2 calculations were performed using the aug cc pvtz basis set and the wb97xd functional. The IEFPCM method was used to model the presence of water. 3

4 Reaction velocity (min -1 ) [S]/V0 Michaelis-Menten and Hanes plots for compounds 5 and 6 15 Hanes-Wolf plot -galactosidase (Asp. Ory.) Deoxy-galactonoeurostegine [ S] 5.3 [ S] 7.1 V No Inhibitor 50 nm 10 5 [ S] 6.5 [ S] 3.0 V [S]/mM Michaelis-Menten plot -galactosidase (Asp. Ory.) Deoxy-galactonoeurostegine No Inhibitor 50 nm [S]/mM 4

5 Reaction velocity (min -1 ) [S]/V Hanes-Wolf plot -glucosidase (Almonds) Deoxy-galactonoeurostegine [ S] 11.3 [ S] 72.2 V No Inhibitor 50 um 100 [ S] 10.2 [ S] 48.5 V [S] Michaelis-Menten plot -glucosidase (Almonds) Deoxy-galactonoeurostegine No Inhibitor 50 um [S] 5

6 Reaction velocity (min -1 ) [S]/V Hanes-Wolf plot -galactosidase (Asp. Ory.) Galactonoeurostegine [ S] 5.3 [ S] 7.1 V No Inhibitor 50 nm 5 [ S] 6.5 [ S] 3.0 V [S]/mM Michaelis-Menten plot -galactosidase (Asp. Ory.) Galactonoeurostegine No Inhibitor 50 nm [S]/mM 6

7 Reaction velocity (min -1 ) Non-competitive - 30 min preincubation Michaelis-Menten plot -glucosidase (Almonds) Galactonoeurostegine 0.15 No Inhibitor 50 nm [S]/mM 7

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