Jan 19, 2026

Public workspaceSynthesis of WUSTL0717

DOI
https://dx.doi.org/10.17504/protocols.io.bp2l6dy2kvqe/v1
  • LINGAIAH MARAM1,
  • Bahaa Elgendy2
  • 1Washington University in Saint Louis, School of Medicine;
  • 2Washington University in St. Louis
LINGAIAH MARAM
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DOI: https://dx.doi.org/10.17504/protocols.io.bp2l6dy2kvqe/v1
Protocol Citation: LINGAIAH MARAM, Bahaa Elgendy 2026. Synthesis of WUSTL0717. protocols.io https://dx.doi.org/10.17504/protocols.io.bp2l6dy2kvqe/v1
License: This is an open access protocol distributed under the terms of the Creative Commons Attribution License,  which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited
Protocol status: Working
We use this protocol and it's working
Created: February 01, 2025
Last Modified: January 19, 2026
Protocol Integer ID: 119400
Keywords: lxr agonist wustl0717 for the treatment, restricted lxr agonist wustl0717, synthesis of wustl0717, short bowel syndrome, synthesis of the gut, wustl0717, mass spectrometry, gut, nmr
Funders Acknowledgements:
National Institute of Health
Grant ID: R01DK119147
Abstract
This protocol outlines the synthesis of the Gut-restricted LXR Agonist WUSTL0717 for the treatment of Short bowel syndrome (SBS), including structural characterization using nuclear magnetic resonance (NMR) and mass spectrometry (LC-MS).
Troubleshooting
Synthesis of WUSTL0717
Lingaiah Maram 1,2,3, Bahaa Elgendy 1,2,3
1Department of Anesthesiology, Washington University in St. Louis, St. Louis, MO, USA.
2Center for Clinical Pharmacology, Washington University School of Medicine and University of Health
Sciences and Pharmacy, St. Louis, Missouri 63110, United States.
3Department of Pharmaceutical and Administrative Sciences, University of Health Sciences and
Pharmacy, St. Louis, Missouri 63110, United States.

Abstract
This protocol outlines the synthesis of the Gut-restricted LXR Agonist WUSTL0717 for the treatment of Short bowel syndrome (SBS), including structural characterization using nuclear magnetic resonance (NMR) and mass spectrometry (LC-MS).
N-(2-Chloro-3-(trifluoromethyl)benzyl)-2,2-diphenylethan-1-aminea
1. Combine 3.0 g of 2,2-diphenylethan-1-amine (15.20 mmol, 1.0 equiv.) and 3.17 g of 2-chloro-3-(trifluoromethyl)benzaldehyde (15.20 mmol, 1.0 equiv.) in 20.0 mL of 1,2-dichloroethane.

2. Add 4.89 g of NaBH(OAc)₃ (23.11 mmol, 1.5 equiv.) and a few drops of acetic acid to the mixture.

3. Stir the reaction at room temperature under a nitrogen atmosphere for 16 hours.

4. Monitor the reaction progress using LC-MS and TLC.

5. Quench the reaction mixture with saturated aqueous NaHCO₃.

6. Extract the product with 1,2-dichloroethane (2 × 100.0 mL).

7. Dry the combined organic layers over anhydrous Na₂SO₄.

8. Filter the solution and concentrate it by evaporating the solvent.

9. Collect the crude product as a colorless oil.

10. Yield: 5.8 g (98%).

11. Characterization:
LC-MS (ESI) m/z: 390.90 [M+H]+

1H NMR (400 MHz, CDCl3): δ 7.62 (dd, J = 7.9, 1.7 Hz, 1H), 7.56 (dd, J = 7.7, 1.6 Hz, 1H), 7.38 – 7.28 (m, 6H), 7.30 –7.18 (m, 8H), 4.25 (t, J = 7.7 Hz, 1H), 3.99 (s, 2H), 3.27 (d, J = 7.7 Hz, 2H).
 
13C{1H} NMR (100 MHz, CDCl3): δ 142.50, 139.91, 133.15, 131.55, 128.93, 128.74, 128.70, 128.65, 128.25, 128.00, 127.95, 126.84, 126.65, 126.42, 126.21 (q, JC,F = 6.0 Hz), 124.30, 121.58, 53.78, 51.15, 51.01.
3-Bromo-N-(2-chloro-3-(trifluoromethyl)benzyl)-N-(2,2-diphenylethyl)propan-1-amine
1. Dissolve 1.0 g of N-(2-Chloro-3-(trifluoromethyl)benzyl)-2,2-diphenylethan-1-amine (2.56 mmol) and 2.61 mL of 1,3-dibromopropane (25.65 mmol) in 5.0 mL of acetonitrile.

2. Add 531.0 mg of K₂CO₃ (3.84 mmol, 1.5 equiv.) to the reaction mixture.
3. Heat the mixture to reflux at 90°C and stir for 15 hours.

4. Cool the reaction mixture to room temperature.

5. Filter the mixture through a pad of Celite and wash with ethyl acetate (EtOAc).

6. Concentrate the filtrate in vacuo to remove EtOAc.

7. Remove excess 1,3-dibromopropane under reduced pressure.

8. Collect the titled product as a nearly pure colorless oil.

9. Yield: 1.18 g (90%).

10. Characterization:
LC-MS (ESI) m/z: 511.95 [M+H]+

1H NMR (400 MHz, CDCl3): δ 7.60 (dd, J = 7.6, 1.8 Hz, 1H), 7.31 (t, J = 7.2 Hz, 6H), 7.28 – 7.19 (m, 8H), 7.17 (t, J = 7.6 Hz, 2H), 4.21 (t, J = 7.7 Hz, 1H), 3.82 (s, 2H), 3.26 (t, J = 6.4 Hz, 2H), 3.18 (d, J = 7.7 Hz, 2H), 2.75 (t, J = 6.4 Hz, 2H), 1.98 (p, J = 6.8 Hz, 2H).

13C NMR (100 MHz, CDCl3): δ 143.24, 139.45, 133.87, 131.50, 128.74, 128.60, 128.37, 128.25, 128.17, 128.11, 128.00, 126.84, 126.38, 126.18, 125.95 (q, JC,F = 5.0 Hz), 124.44, 121.66, 60.48, 56.21, 52.68, 49.74, 31.90, 30.39.
Ethyl 2-(3-(3-((2-chloro-3-(trifluoromethyl)benzyl)(2,2- diphenylethyl)amino)propoxy)phenyl)acetate
1. Dissolve 1.65 g of (3-Bromo-propyl)-(2-chloro-3-trifluoromethyl-benzyl)-diphenylethyl-amine (3.23 mmol) and 640.0 mg of ethyl 2-(3-hydroxyphenyl)acetate (3.55 mmol) in 10.0 mL of acetonitrile.

2. Add 680.0 mg of K₂CO₃ (4.90 mmol, 1.5 equiv.) to the solution.

3. Heat the reaction mixture to reflux at 90°C and stir for 20 hours.

4. Cool the reaction mixture to room temperature.

5. Filter through a pad of Celite and wash with ethyl acetate (EtOAc).

6. Concentrate the filtrate in vacuo to remove the solvent.

7. Purify the crude product by silica gel column chromatography using EtOAc/hexane (1:4) as the eluent.

8. Collect the title compound as a colorless oil.

9. Yield: 1.54 g (78%).

10.Characterization:
LC-MS (ESI) m/z: 611.20 [M+H]+

1H NMR (400 MHz, CDCl3): δ 7.46 (dd, J = 7.9, 1.6 Hz, 1H), 7.30 – 7.12 (m, 12H), 6.95 – 6.83 (m, 2H), 6.68 (t, J = 2.1 Hz, 1H), 6.64 (dd, J = 8.1, 2.5 Hz, 1H), 4.25 – 4.05 (m, 3H), 3.80 (s, 2H), 3.68 (t, J = 5.9 Hz, 2H), 3.58 (s, 2H), 3.15 (d, J = 7.7 Hz, 2H), 2.71 (t, J = 6.5 Hz, 2H), 1.85 (p, J = 6.3 Hz, 2H), 1.26 (t, J = 7.1 Hz, 3H).

13C NMR (100 MHz, CDCl3): δ 171.48, 158.94, 143.39, 139.65, 135.41, 133.76, 129.40, 128.33, 128.20, 126.31, 126.12, 125.71 (q, JC,F = 6.0 Hz), 121.38, 115.35, 112.98, 64.87, 60.84, 60.28, 56.00, 50.76, 49.72, 41.45, 26.81, 14.18.
2-(3-(3-((2-Chloro-3-(trifluoromethyl)benzyl)(2,2- diphenylethyl)amino)propoxy)phenyl)acetic acid (GW3965):
1. Dissolve 1.2 g of Ethyl 2-(3-(3-((2-chloro-3-(trifluoromethyl)benzyl)(2,2- diphenylethyl)amino)propoxy)phenyl)acetate (1.96 mmol) in a 1:1 mixture of THF and H₂O.

2. Add 490.0 mg of LiOH·H₂O (11.80 mmol) at room temperature.

3. Stir the reaction mixture at room temperature for 20 hours, monitoring progress by LC-MS and TLC analysis.

4. Dilute the reaction mixture with 10.0 mL of ethyl acetate (EtOAc).

5. Carefully neutralize with 1N aqueous HCl.

6. Extract the mixture with EtOAc (3 × 20.0 mL).

7. Wash the combined organic layers with water and brine.

8. Dry over anhydrous Na₂SO₄, filter, and concentrate under reduced pressure.

9. Collect the crude product GW3965 as a colorless oil.

10. Yield: 1.0 g (90%), with >99% purity confirmed by HPLC area percentage.

11. Characterization:
LC-MS(ESI) m/z = 583.10 [M+H]+.

1H NMR (400 MHz, CDCl3): δ 7.45 (d, J = 7.8 Hz, 1H), 7.31 – 7.07 (m, 13H), 6.86 (d, J = 7.4 Hz, 2H), 6.65 (dd, J = 9.1, 1.8 Hz, 2H), 4.22 – 4.05 (m, 1H), 3.79 (s, 2H), 3.64 (d, J = 12.5 Hz, 4H), 3.15 (d, J = 7.7 Hz, 2H), 2.77 – 2.62 (m, 2H), 1.84 (d, J = 6.2 Hz, 2H).

13C NMR (100 MHz, CDCl3): δ 174.24, 158.97, 143.36, 139.60, 136.10, 134.47, 133.73, 129.52, 128.34, 128.18, 126.32, 126.12, 125.76, 125.70, 121.47, 115.44, 113.23, 64.81, 60.20, 55.96, 50.66, 49.67, 40.99, 26.75
2-(3-(3-((2-Chloro-3-(trifluoromethyl)benzyl)(2,2-diphenylethyl)amino)propoxy)phenyl)acetamide (WUSTL0717)b
1. Dissolve GW3965 (0.55 g, 0.945 mmol) in 10.0 mL of acetonitrile (CH₃CN) under a nitrogen atmosphere.

2. Add triethylamine (Et₃N, 0.221 g, 0.3 mL, 2.192 mmol, 2.32 equiv.) to the solution.

3. Add Py.BOP (0.57 g, 1.096 mmol, 1.16 equiv.) at room temperature.

4. Stir the reaction mixture for 10 minutes at room temperature.

5. Add NH₃ in dioxane (0.4 M, 2.83 mL, 1.2 equiv.) to the reaction mixture.

6. Stir the reaction mixture at room temperature for 4 hours.

7. Concentrate the mixture under reduced pressure.

8. Partition the residue between EtOAc (20.0 mL) and 5% aqueous NaHCO₃ solution.

9. Separate the organic layer, dry over anhydrous Na₂SO₄, and concentrate.

10.Purify the crude product by flash column chromatography using EtOAc/Hexane (1:1).

11. Collect the titled product as a colorless oil.

12. Yield: 428.3 mg (78%).

13. Characterization:
LC-MS(ESI) m/z = 582.08 [M+H]+.

1H NMR (400 MHz, CDCl3): δ 7.48 (dd, J = 7.8, 1.7 Hz, 1H), 7.29 – 7.09 (m, 13H), 6.93 (t, J = 7.8 Hz, 1H), 6.85 (d, J = 7.5 Hz, 1H), 6.73 – 6.60 (m, 2H), 5.55 (s, 1H), 5.39 (s, 1H), 4.13 (q, J = 7.8 Hz, 1H), 3.79 (s, 2H), 3.70 (t, J = 6.0 Hz, 2H), 3.55 (s, 2H), 3.15 (d, J = 7.7 Hz, 2H), 2.72 (t, J = 6.5 Hz, 2H), 1.87 (dd, J = 12.8, 6.6 Hz, 2H).

13C NMR (100 MHz, CDCl3): δ 173.24, 159.33, 143.33, 139.62, 136.15, 133.81, 131.36, 130.03, 128.73, 128.33, 128.18, 128.00, 126.83, 126.33, 126.05, 127.78 (d, JC,F = 6.0 Hz), 121.43, 115.39, 113.36, 65.05, 60.26, 56.07, 50.72, 49.69, 43.39, 26.77


WUSTL0717. HCl:
1. Dissolve 500.0 mg (0.86 mmol) of 2-(3-(3-((2-chloro-3-(trifluoromethyl)benzyl)(2,2-diphenylethyl)amino)propoxy)phenyl)acetamide in ether.
2. Add 0.86 mL of 2M HCl in ether to the solution at room temperature.
3. Stir the mixture at room temperature for 1 hour.
4. Evaporate the solvent under reduced pressure.
5. Collect the HCl salt as a colorless solid.
6. Yield: 510.0 mg (96%).
Protocol references
(a). A. F. Abdel-Magid, K. G. Carson, B. D. Harris, C. A. Maryanoff and R. D. Shah, J .Org .Chem., 1996, 61, 3849–3862.
(b) D. G. Washburn, T. H. Hoang, N. Campobasso, A. Smallwood, D. J. Parks, C. L. Webb, K. A. Frank, M. Nord, C. Duraiswami, C. Evans, M. Jaye and S. K. Thompson, Bioorg. Med. Chem. Lett., 2009, 19, 1097–1100.