2-[2-(2-azidoethoxy)ethoxy]ethanol

Conditions: temp:0.00; pressure:0.00; stir:

Conditions: temp:0.00; pressure:0.00; stir:; See reaction.notes.procedure_details.

Procedure: To a solution in which the 2-(2-(2-azidoethoxy)ethoxy)ethanol (3.05 g, 17.40 mmol) compound of Chemical Formula 8 was dissolved in methylene chloride (15 mL), 4-dimethylaminopyridine (DMAP) (212.6 mg, 1.74 mmol), triethylamine (TEA) (7.28 mL, 52.20 mmol) and tosyl chloride (3.98 g, 20.88 mmol) were added consecutively at room temperature. The post-reaction mixture obtained after stirring the reaction mixture for 12 hours at room temperature was partitioned to distilled water (40 mL) and ethyl acetate (120 mL). The organic layer was separated, washed with saturated salted water (15 mL), dried with anhydrous sodium sulfate, and vacuum distilled to give 2-(2-(2-azidoethoxy)ethoxy)ethyl 4-methylbenzenesulfonate (5.15 g, 90%). 1H NMR (500 MHz, CDCl3) δ 7.79 (d, 2H, J=8.2 Hz), 7.33 (d, 2H, J=8.2 Hz), 4.15 (t, 2H, J=5.0 Hz), 3.69 (t, 2H, J=5.0 Hz), 3.63 (t, 2H, J=5.5 Hz), 3.59 (s, 4H), 3.36 (t, 2H, J=5.0 Hz), 2.44 (s, 3H); 13C NMR (125 MHz, CDCl3) δ 144.9, 133.0, 129.9 (2), 128.0 (2), 70.8, 70.7, 70.1, 69.3, 68.8, 50.7, 21.7. The post-reaction mixture obtained was partitioned to distilled water (40 mL) and ethyl acetate (120 mL) The organic layer was separated washed with saturated salted water (15 mL) dried with anhydrous sodium sulfate, and vacuum distilled

Conditions: temp:100.01; pressure:0.00; stir:; See reaction.notes.procedure_details.

Procedure: To a solution in which 2-(2-(2-chloroethoxy)ethoxy)ethanol (3.48 g, 20.63 mmol) of Chemical Formula 7 was dissolved in DMF (15 mL), sodium azide (4.02 g, 61.89 mmol) was added at room temperature, and the mixture was heated to 100° C. and stirred for 12 hours. After the reaction, DMF was removed by vacuum distillation, and the residue was partitioned to distilled water (40 mL) and ethyl acetate (150 mL). The organic layer was separated, washed with saturated salted water (15 mL), dried with anhydrous sodium sulfate, and vacuum distilled, and the residue obtained was column chromatographed on silica gel (hexane/ethyl acetate (2:1)) to give 2-(2-(2-azidoethoxy)ethoxy)ethanol (3.05 g, 84%). 1H NMR (500 MHz, CDCl3) δ 3.72-3.74 (m, 2H), 3.65-3.69 (m, 6H), 3.60-3.62 (m, 2H), 3.39 (t, 2H, J=5.0 Hz). was removed by vacuum distillation the residue was partitioned to distilled water (40 mL) and ethyl acetate (150 mL) The organic layer was separated washed with saturated salted water (15 mL) dried with anhydrous sodium sulfate, and vacuum distilled the residue obtained chromatographed on silica gel (hexane/ethyl acetate (2:1))

Conditions: temp:0.00; pressure:0.00; stir:; See reaction.notes.procedure_details.

Procedure: This protocol was modified to synthesis 5β exclusively. Briefly, compound 2 (1.0 g, 3.03 mmol) and alcohol 4 (1.60 g, 9.11 mmol) were dissolved in anhydrous DCM (20 mL), followed by addition of dried 4 A° MS (0.4 g) and reaction mixture stirred at room temperature for 20 minutes under N2. CSA (camphor sulphonic acid, 0.615 g, 9.0 mL, 9.1 mmol) was added at room temperature and 8 hours, followed by heating to 45° C. for 24 hours. The reaction mixture was cooled to room temperature, neutralized with Et3N (0.2 mL, pH 7.5), diluted with DCM (100 mL) and extracted using saturated NaHCO3 solution. The organic layer was washed with water, brine and dried over Na2SO4. Solvents were evaporated under reduced pressure and the residue purified by silica gel column chromatography using DCM:EtOAc:MeOH (8.0:1.5:0.5) to obtain compound 5β (1.15 g, 85%). The spectral data matched with above compound 5β. This protocol was modified to synthesis 5β exclusively followed by addition of dried 4 A° MS (0.4 g) and reaction mixture by heating to 45° C. for 24 hours The reaction mixture was cooled to room temperature extracted The organic layer was washed with water, brine dried over Na2SO4 Solvents were evaporated under reduced pressure the residue purified by silica gel column chromatography to obtain compound 5β (1.15 g, 85%)

Conditions: temp:80.01; pressure:0.00; stir:; See reaction.notes.procedure_details.

Procedure: Compound 3 (10.0 g, 59.50 mmol) was dissolved in anhydrous DMF (200 mL), to which NaN3 (77.3 g, 119.00 mmol) was added and reaction mixture stirred at 80° C. or 48 hours. Solvents were removed under reduced pressure and residue dissolved in DCM (200 mL), washed with water (400 mL) and brine (100 mL) before being dried over Na2SO4. Solvent was evaporated under reduced pressures and residue purified by silica gel column chromatography using EA:hexane (6:4) to obtain 4 as a liquid (7.80 g, 75% yield). 1H-NMR (500 MHz, CDCl3): δ 2.34 (d, 1H, J=5.5 Hz, OH), 3.93 (t, 2H, J=6.0 Hz, Ha), 3.61 (t, 2H, J=4.5 Hz, Hb), 3.65-3.68 (m, 6H, Hc, He, Hf), 3.73 (q, 2H, J=5.5 Hz, Hd); 13C-NMR (125 MHz, CDCl3): δ 50.62 (Ca), 61.73 (Cf), 70.02 (Cb), 70.36 (Ce), 70.63 (Cc), 72.45 (Cd). EI-MS: [M+Na]+ C6H13N3NaO3 calcd 198.08, obsd 198.07 was added reaction mixture Solvents were removed under reduced pressure and residue dissolved in DCM (200 mL) washed with water (400 mL) and brine (100 mL) before being dried over Na2SO4 Solvent was evaporated under reduced pressures and residue purified by silica gel column chromatography

Conditions: temp:80.01; pressure:0.00; stir:; See reaction.notes.procedure_details.

Procedure: Compound 3 (10.0 g, 59.50 mmol) was dissolved in anhydrous DMF (200 mL), to which NaN3 (77.3 g, 119.00 mmol) was added and reaction mixture stirred at 80° C. or 48 hours. Solvents were removed under reduced pressure and residue dissolved in DCM (200 mL), washed with water (400 mL) and brine (100 mL) before being dried over Na2SO4. Solvent was evaporated under reduced pressures and residue purified by silica gel column chromatography using EA:hexane (6:4) to obtain 4 as a liquid (7.80 g, 75% yield). 1H-NMR (500 MHz, CDCl3): δ 2.34 (d, 1H, J=5.5 Hz, OH), 3.93 (t, 2H, J=6.0 Hz, Ha), 3.61 (t, 2H, J=4.5 Hz, Hb), 3.65-3.68 (m, 6H, Hc, He, Hf), 3.73 (q, 2H, J=5.5 Hz, Hd); 13C-NMR (125 MHz, CDCl3): δ 50.62 (Ca), 61.73 (Cf), 70.02 (Cb), 70.36 (Ce), 70.63 (Cc), 72.45 (Cd). EI-MS: [M+Na]+ C6H13N3NaO3 calcd 198.08, obsd 198.07 was added reaction mixture Solvents were removed under reduced pressure and residue dissolved in DCM (200 mL) washed with water (400 mL) and brine (100 mL) before being dried over Na2SO4 Solvent was evaporated under reduced pressures and residue purified by silica gel column chromatography

Conditions: temp:0.00; pressure:0.00; stir:; See reaction.notes.procedure_details.

Procedure: To a stirred solution of Mono-Propargyl Curcumin 5 (63 mg, 0.16 mmol) and 2-(2-(2-azidoethoxy)ethoxy)ethanol (46 mg, 0.26 mmol) in tBuOH (1.1 mL) and CHCl3 (0.3 mL) was added a prepared solution of Cu(OAc)2 (8 mg, 0.03 mmol) and sodium ascorbate (13 mg, 0.07 mmol) in H2O (1.3 mL). After vigorous stirring overnight the solvent was removed under vacuum. The mixture was dissolved in CHCl3, washed with H2O and the organic phase was separated, dried over Na2SO4 and evaporated. Purification was performed by column chromatography, eluting with CH2Cl2-CH2Cl2: MeOH 98:2. Yield 26%. NMR NMR 1H (CDCl3), δ (ppm): 3.55-3.60 (m, 8H); 3.72-3.72 (t, 3H); 3.87-3.94 (m, 8H); 4.54-4.55 (t, 2H); 5.80 (s, 1H); 6.07 (s, 1H); 6.46-6.50 (dd, 2H); 6.92-6.94 (d, 1H); 7.05-7.12 (m, 5H); 7.56-7.60 (q, 2H); 7.92 (s, 1H).). MS (ESI) calcd. for C30H35N3O9: 581.61. Found: 582.3 [M+H]+, 604.3 [M++Na], 620.3 [M++K]. was removed under vacuum The mixture was dissolved in CHCl3 washed with H2O the organic phase was separated dried over Na2SO4 evaporated Purification eluting with CH2Cl2-CH2Cl2

Conditions: temp:0.00; pressure:0.00; stir:; See reaction.notes.procedure_details.

Procedure: To a stirred solution of Mono-Propargyl Curcumin 5 (63 mg, 0.16 mmol) and 2-(2-(2-azidoethoxy)ethoxy)ethanol (46 mg, 0.26 mmol) in tBuOH (1.1 mL) and CHCl3 (0.3 mL) was added a prepared solution of Cu(OAc)2 (8 mg, 0.03 mmol) and sodium ascorbate (13 mg, 0.07 mmol) in H2O (1.3 mL). After vigorous stirring overnight the solvent was removed under vacuum. The mixture was dissolved in CHCl3, washed with H2O and the organic phase was separated, dried over Na2SO4 and evaporated. Purification was performed by column chromatography, eluting with CH2Cl2-CH2Cl2: MeOH 98:2. Yield 26%. NMR NMR 1H (CDCl3), δ (ppm): 3.55-3.60 (m, 8H); 3.72-3.72 (t, 3H); 3.87-3.94 (m, 8H); 4.54-4.55 (t, 2H); 5.80 (s, 1H); 6.07 (s, 1H); 6.46-6.50 (dd, 2H); 6.92-6.94 (d, 1H); 7.05-7.12 (m, 5H); 7.56-7.60 (q, 2H); 7.92 (s, 1H).). MS (ESI) calcd. for C30H35N3O9: 581.61. Found: 582.3 [M+H]+, 604.3 [M++Na], 620.3 [M++K]. was removed under vacuum The mixture was dissolved in CHCl3 washed with H2O the organic phase was separated dried over Na2SO4 evaporated Purification eluting with CH2Cl2-CH2Cl2

Conditions: temp:0.00; pressure:0.00; stir:; See reaction.notes.procedure_details.

Procedure: To a stirred solution of Mono-Propargyl Curcumin 5 (63 mg, 0.16 mmol) and 2-(2-(2-azidoethoxy)ethoxy)ethanol (46 mg, 0.26 mmol) in tBuOH (1.1 mL) and CHCl3 (0.3 mL) was added a prepared solution of Cu(OAc)2 (8 mg, 0.03 mmol) and sodium ascorbate (13 mg, 0.07 mmol) in H2O (1.3 mL). After vigorous stirring overnight the solvent was removed under vacuum. The mixture was dissolved in CHCl3, washed with H2O and the organic phase was separated, dried over Na2SO4 and evaporated. Purification was performed by column chromatography, eluting with CH2Cl2—CH2Cl2: MeOH 98:2. Yield 26%. NMR 1H (CDCl3), δ (ppm): 3.55-3.60 (m, 8H); 3.72-3.72 (t, 3H); 3.87-3.94 (m, 8H); 4.54-4.55 (t, 2H); 5.80 (s, 1H); 6.07 (s, 1H); 6.46-6.50 (dd, 2H); 6.92-6.94 (d, 1H); 7.05-7.12 (m, 5H); 7.56-7.60 (q, 2H); 7.92 (s, 1H).). MS (ESI) calcd. for C30H35N3O9: 581.61. found: 582.3 [M+H]+, 604.3 [M++Na], 620.3 [M++K]. was removed under vacuum The mixture was dissolved in CHCl3 washed with H2O the organic phase was separated dried over Na2SO4 evaporated Purification eluting with CH2Cl2—CH2Cl2

Conditions: temp:0.00; pressure:0.00; stir:; See reaction.notes.procedure_details.

Procedure: To a stirred solution of Mono-Propargyl Curcumin 5 (63 mg, 0.16 mmol) and 2-(2-(2-azidoethoxy)ethoxy)ethanol (46 mg, 0.26 mmol) in tBuOH (1.1 mL) and CHCl3 (0.3 mL) was added a prepared solution of Cu(OAc)2 (8 mg, 0.03 mmol) and sodium ascorbate (13 mg, 0.07 mmol) in H2O (1.3 mL). After vigorous stirring overnight the solvent was removed under vacuum. The mixture was dissolved in CHCl3, washed with H2O and the organic phase was separated, dried over Na2SO4 and evaporated. Purification was performed by column chromatography, eluting with CH2Cl2—CH2Cl2: MeOH 98:2. Yield 26%. NMR 1H (CDCl3), δ (ppm): 3.55-3.60 (m, 8H); 3.72-3.72 (t, 3H); 3.87-3.94 (m, 8H); 4.54-4.55 (t, 2H); 5.80 (s, 1H); 6.07 (s, 1H); 6.46-6.50 (dd, 2H); 6.92-6.94 (d, 1H); 7.05-7.12 (m, 5H); 7.56-7.60 (q, 2H); 7.92 (s, 1H).). MS (ESI) calcd. for C30H35N3O9: 581.61. found: 582.3 [M+H]+, 604.3 [M++Na], 620.3 [M++K]. was removed under vacuum The mixture was dissolved in CHCl3 washed with H2O the organic phase was separated dried over Na2SO4 evaporated Purification eluting with CH2Cl2—CH2Cl2

Conditions: temp:0.00; pressure:0.00; stir:; See reaction.notes.procedure_details.

Procedure: 8-Trifluoroacetamido-3,6-dioxa-1-octanol (1f: 8-azido-3,6-dioxa-1-octanol (Amvamzollo, P. H.; Sinay, P. Carbohydr. Res. 1986, 150, 199-212) (0.7 g, 4.0 mmol) was synthesized from triethylene glycol following the reported procedure (Bertozzi, C. R.; Bednarski, M. D. J. Org. Chem. 1991, 56, 4326-4329). The azide was combined with Pd/C (150 mg) in MeOH (8.0 mL) and stirred 12 h under H2 (1 atm). The suspension was filtered over celite, and the filtrate was concentrated to afford the amine (0.590 g, 4.0 mmol) in quantitative yields (Sato, H.; Hayashi, E.; Yamada, N.; Yatagai, M.; Takahara, Y. Bioconjugate Chem. 2001, 12, 701-710.). Following general procedure 1, 2-[2-(2-aminoethoxy)ethoxy]ethanol (0.590 g, 3.95 mmol) was combined with triethylamine (1.378 mL, 9.89 mmol) and trifluoroacetic anhydride (0.782 mL, 5.54 mmol) in MeOH (5 mL). Purification by silica gel chromatography (EtOAc) yielded 0.576 g (60%) of if as a white solid. 1H (300 MHz, CD3OD): δ 7.87 (br s, 1H), 3.73 (m, 2H), 3.68-3.60 (m, 10H), 3.55 (t, 2H, J=4.5 Hz), 3.32 (br s, 1H); 13C (75 mHz, CD3OD): δ 157.6 (q, J=36.6 Hz), 116.1 (q, J=286.1 Hz), 72.68, 70.35, 70.24, 68.90, 61.52, 39.84; ESI-MS calcd for C8H14F3NO4 [M−H]−: 244.0797. found 244.0795. The suspension was filtered over celite the filtrate was concentrated

Conditions: temp:0.00; pressure:0.00; stir:; See reaction.notes.procedure_details.

Procedure: 8-Trifluoroacetamido-3,6-dioxa-1-octanol (1f: 8-azido-3,6-dioxa-1-octanol (Amvamzollo, P. H.; Sinay, P. Carbohydr. Res. 1986, 150, 199-212) (0.7 g, 4.0 mmol) was synthesized from triethylene glycol following the reported procedure (Bertozzi, C. R.; Bednarski, M. D. J. Org. Chem. 1991, 56, 4326-4329). The azide was combined with Pd/C (150 mg) in MeOH (8.0 mL) and stirred 12 h under H2 (1 atm). The suspension was filtered over celite, and the filtrate was concentrated to afford the amine (0.590 g, 4.0 mmol) in quantitative yields (Sato, H.; Hayashi, E.; Yamada, N.; Yatagai, M.; Takahara, Y. Bioconjugate Chem. 2001, 12, 701-710.). Following general procedure 1, 2-[2-(2-aminoethoxy)ethoxy]ethanol (0.590 g, 3.95 mmol) was combined with triethylamine (1.378 mL, 9.89 mmol) and trifluoroacetic anhydride (0.782 mL, 5.54 mmol) in MeOH (5 mL). Purification by silica gel chromatography (EtOAc) yielded 0.576 g (60%) of if as a white solid. 1H (300 MHz, CD3OD): δ 7.87 (br s, 1H), 3.73 (m, 2H), 3.68-3.60 (m, 10H), 3.55 (t, 2H, J=4.5 Hz), 3.32 (br s, 1H); 13C (75 mHz, CD3OD): δ 157.6 (q, J=36.6 Hz), 116.1 (q, J=286.1 Hz), 72.68, 70.35, 70.24, 68.90, 61.52, 39.84; ESI-MS calcd for C8H14F3NO4 [M−H]−: 244.0797. found 244.0795. The suspension was filtered over celite the filtrate was concentrated