Contents
Scheme 2.4. The design concept of the dual emitting nucleoside
2.9. Experimental Section 1. General Experimental
2.9.2. Synthesis and Characterizations
Synthesis of 4-iodo-N-methylaniline (2.37): This compound was synthesized following a modified literature procedure.153 3.95 g (13.84 mmol) of N-methylaniline (2.36) was taken in a round bottom (R.B.) flask and was dissolved in 20 ml of pyridine: dioxane (1:1). The resulting reaction mixture was cooled to 0 °C in an ice bath and half of the total 5.6 g (22.14 mmol) of I2 was added to it. While maintaining the ice cooled condition, remaining half amount of I2 was added after 2 hours. After stirring for about 3 hours, the ice bath was removed and the reaction mixture was stirred at room temperature for another 22 hours. After completion of the reaction, monitored by TLC, the reaction mixture was partitioned between ethyl acetate and water. The collected organic layer was washed with saturated sodium thiosulfate solution, water, and brine solution. After evaporation, the crude product was purified by column chromatography (Si-gel, Hex: EtOAc = 30:1) to obtain the 4- iodo-N-methylaniline 2.37 as a dark yellow liquid. Yield 92 % (7.97 g).1H NMR (CDCl3, 400 MHz): 7.4 (d, J = 8.4 Hz, 2H), 6.36 (d, J = 8.4 Hz, 2H), 3.71 (s, 1H), 2.767 (s, 3H); 13C NMR (CDCl3, 100MHz): 148.9, 137.8, 114.7, 77.7, 30.6.
Synthesis of N-methyl-4-((trimethylsilyl)ethynyl)aniline (2.38): This compound was synthesized following a modified literature procedure. 5 g (21.45 mmol) of 4-iodo-N-methylaniline (2.37) was taken in a dry R.B.
and dissolved in 20 ml of dry benzene. To the above solution, 10 ml of dry n-butylamine was added and the resulting mixture was degassed for 10 minutes by bubbling N2 through it.
PdCl2(PPh3)2 (0.45 g, 0.03 mmol) followed by CuI (0.04 g, 0.01 mmol), were added to the reaction mixture while continuing the degassing. Finally, trimethylsilylacetylene (3.16 g, 32.17 mmol) was added to the reaction mixture and it was refluxed (70-80 °C) under N2 atmosphere for 7 hours. After completion of the reaction, the reaction mixture was partitioned between ethyl acetate and water. The collected organic layer was washed with an aqueous ammonium chloride solution, water, brine solution and dried over anhydrous Na2SO4. After evaporation, the crude product was purified by column chromatography (Si-gel, Hex: EtOAc = 30:1) to obtain product 2.38 as a reddish-brown liquid. Yield: 83% (3.62 g). IR (KBr): ̃ 3421, 2147, 1609, 1521cm-1; 1H NMR (CDCl3, 400 MHz): 7.06 (d, J = 8.8 Hz, 2H), 6.23 (d, J = 8.8 Hz, 2H), 3.64 (s, 1H), 2.56 (s, 3H), 0.00 (s, 9H); 13C NMR (CDCl3, 100 MHz): 149.5, 133.4, 111.8, 106.6, 91.1, 30.4, 0.7;+ESI-HRMS calculated for C12H18NSi [M+H]+ 204.1203, found 204.1198.
Synthesis of N-methyl-N-(prop-2-yn-1-yl)-4-((trimethylsilyl)ethynyl)aniline (2.39): This compound was synthesized following a modified literature procedure.154 2.5 g (12.29 mmol) of N-methyl-4- ((trimethylsilyl)ethynyl)aniline (2.38) was taken in a R.B. and dissolved in 8 ml of dry DMF. K2CO3 (2.55 g, 18.44 mmol) was added to the mixture followed by propargylbromide (2.19 g, 18.44 mmol). The resulting reaction mixture was stirred at 50 °C for 15 hours under N2 atmosphere. After completion of reaction, the reaction mixture was partitioned between ethyl acetate and water. Collected organic layer was washed with water, brine solution and dried over anhydrous Na2SO4. After evaporation, the product was purified by column chromatography (Si-gel, Hex: EtOAc = 30:1) and obtained as brown semi solid. Yield: 79 %( 2.36g). IR (KBr): ̃ 3279, 2147, 1883, 1605, 1517cm-1; 1H NMR (CDCl3, 400 MHz): 7.36 (d, J = 8.4 Hz, 2H), 6.71 (d, J = 8.4 Hz, 2H), 4.05 (d, J = 1.6 Hz, 2H), 2.98 (s, 3H), 2.17 (s, 2H), 0.2 (s, 9H); 13C NMR (CDCl3, 100 MHz):
148.9, 133.3, 113.4, 106.2, 91.9, 79.0, 72.3, 42.2, 38.5, 0.3; +ESI-HRMS calculated for C15H20NSi [M+H]+ 242.1360, found 242.1356.
Synthesis of 3′,5′-di-O-tert-butyldimethylsilyl-5-iodo-2′-deoxyuridine (2.41):This compound was synthesized following a modified literature procedure.155 4 g (11.23 mmol) of 5-iodo-2'-deoxyuridine (2.40) was taken in a dry R.B. and dissolved in dry DMF. The reaction mixture was vacuumed and filled with N2. Imidazole (3.85 g, 56.49 mmol) was added to the reaction mixture and the clear reaction mixture was vacuumed one more time and filled with N2. The above mixture was cooled to 0
°C in an ice bath and t-butyldimethylsilylchloride (5.08 g, 33.69 mmol) was added to the reaction mixture. The ice bath was removed after 1 hour and the resulting mixture was stirred at room temperature under N2 for 18 hours. After completion of reaction, the reaction mixture was partitioned between ethyl acetate and water. Collected organic layer was washed with water, brine solution and dried over anhydrous Na2SO4. After evaporation, the product was purified by column chromatography (Si-gel, Hex: EtOAc = 5:1) and obtained as white foam.
Yield: 96% (6.35 g). IR (KBr): ̃ 3457, 3184, 3062, 2954, 2931, 2857,1694, 1607cm-1; 1H NMR (CDCl3, 600 MHz): 8.96 (s, 1H), 8.07 (s, 1H), 6.26 (t, J = 6.6 Hz, 1H), 4.38 (s, 1H), 3.97 (s, 1H), 3.88 (d, J = 11.4 Hz, 1H), 3.75 (d, J = 10.8 Hz, 1H), 2.31-2.28 (m, 1H), 2.00-1.96 (m, 1H), 0.93 (s, 9H), 0.88 (s, 9H), 0.14 (d, J = 6.6 Hz, 6H), (0.07) (d, J = 4.8 Hz, 6H); 13C NMR (CDCl3, 150 MHz): 160.2, 150.0, 144.6, 88.6, 86.0, 72.7, 68.5, 63.2, 42.2, 38.7, 26.3,
25.9, 18.7, 18.2, -4.4, -4.6, -4.9, -5.0; +ESI-HRMS calculated for C21H40IN2O5Si2 [M+H] + 583.1515, found 583.1511.
Synthesis of 3′,5′-di-O-tert-butyldimethylsilyl-5-(3-((4-
ethynylphenyl)(methyl)amino)propynyl)-2'-deoxyuridine (2.42):This compound was synthesized following a modified literature procedure.156 1.12 g (1.92 mmol) of 3',5'-bis-O-
tert-butyldimethylsilyl-5-iodo-2'-deoxyuridine (2.41) and N-methyl-N-(prop-2-yn-1-yl)-4- ((trimethylsilyl)ethynyl)aniline (2.39) were taken in a dry R.B. and dissolved in 15 ml dry Et3N. The resulting mixture was degassed by bubbling N2 through it. After 10 minutes, PdCl2(PPh3)2
(40.49 mg, 0.057 mmol) followed by CuI (3.66 mg, 0.019 mmol) were added to the above mixture while continuing the degassing. The resulting mixture was stirred at 55 °C for 7 hours.
After completion, the reaction mixture was partitioned between ethyl acetate and water.
Collected organic layer was washed with aqueous ammonium chloride, water, brine solution and dried over anhydrous Na2SO4. After evaporation, the product was purified by column chromatography (Si-gel, Hex: EtOAc = 3:1) and obtained as light brown foam. Yield: 78 % (783 mg). IR (KBr): ̃ 3451, 3047, 2954, 2930, 1714, 1688, 1609 cm-1; 1H NMR (CDCl3, 600 MHz): δ 8.53 (s, 1H), 7.86 (s, 1H), 7.35 – 7.32 (m, 2H), 6.72 – 6.69 (m, 2H), 6.25 (dd, J = 7.5, 5.4 Hz, 1H), 4.38-4.36 (m, 1H), 4.25 (s, 2H), 3.96 (q, J = 2.4 Hz, 1H), 3.84 (dd, J = 11.4, 2.4 Hz, 1H), 3.73 (dd, J = 11.4, 2.4 Hz, 1H), 3.01 (s, 3H), 2.31 – 2.27 (m, 1H), 2.00 – 1.96 (m, 1H), 0.88 (s, 9H), 0.87 (s, 9H), 0.22 (s, 9H), 0.07 (s, 3H), 0.06 (s, 6H), 0.04 (s, 3H); 13C NMR (CDCl3, 150 MHz): δ 161.5, 149.2, 148.9, 142.4, 133.3, 113.1, 111.7, 106.3, 99.8, 91.7, 89.7, 88.5, 86.0, 77.2, 75.3, 72.6, 63.1, 43.0, 42.1, 38.4, 26.1, 25.8, 18.5, 18.1, 0.3, -4.5, -4.7, -5.2, - 5.5; +ESI-HRMS calculated for C36H58N3O5Si3 [M+H] + 696.3679, found 696.3672.
Synthesis of 3′,5′-di-O-tert-butyldimethylsilyl-5-(3-((4-
ethynylphenyl)(methyl)amino)propynyl)-2'-deoxyuridine(2.43): This compound was synthesised following a modified literature procedure.157 991 mg (1.42 mmol) of (2.42) was taken in a dry R.B. and dissolved in 10 ml dry methanol. Anhydrous K2CO3
(984g, 7.11 mmol) was added to the above solution and the resulting mixture was stirred for 4 hours at room temperature. After completion of reaction, the reaction mixture was partitioned between ethyl
acetate and water. Collected organic layer was washed with water, brine solution and dried over anhydrous Na2SO4. After evaporation, the product was purified by column chromatography (Si-gel, Hex: EtOAc = 4:1) and obtained as dark orange foam. Yield: 87%
(773g). IR (KBr): ̃ 3312, 2954, 2928, 1712, 1685, 1609 cm-1; 1H NMR (CDCl3, 600 MHz):
9.23 (s, 1H), 7.86 (s, 1H), 7.36 (d, J = 8.4 Hz, 2H), 6.72 (d, J = 9.0 Hz, 2H), 6.25 (t, J = 6.6 Hz, 2H), 4.36 (t, J = 2.4 Hz, 1H), 3.95 (s, 1H), 3.83 (d, J = 11.4 Hz, 1H), 3.72 (d, J = 11.4, 1H), 3.01 (s, 3H), 2.96 (s, 1H), 2.31-2.28 (m, 1H), 2.00-1.92 (m, 1H), 0.87 (s, 9H), 0.86 (s, 9H), 0.06 (s, 6H), 0.03 (s, 6H); +APCI-HRMS calculated for C33H50N3O5Si2 [M+H]+ 624.3284, found 624.3299.
General Procedure for the Synthesis of Aryl Azides: An ice cold solution of sodium nitrite (3 eqv.) in water was added dropwise to a cold solution of arylamine (1 eqv.) in water and concentrated hydrochloric acid at 0 °C over 7 to10 min. The reaction mixture was slowly stirred for 1-2 min before an ice cold solution of sodium azide (6 eqv.) in water was added dropwise at 0 °C over 10 min. The mixture was stirred for 15 min. The resulting mixture was extracted with hexane. The organic layer was washed with water, followed by a brine solution, dried over anhydrous Na2SO4. After evaporation, the product was passed through a section of silica gel (60-120 mesh). Formation of the azides were confirmed from FT-IR study and yields were within 60%-80% in all cases. The produced azides were then immediately used for the next step without further purification.
General Procedure for Click Reaction: Alkyne (1 eqv.) and azide (1.5 eqv.) were suspended in a 1:1 water/tert-butanol mixture. Sodium ascorbate (0.05 eqv., freshly prepared in 1ml water) was added, followed by CuSO4.5H2O (0.2eqv., freshly prepared in 1ml water).
The reaction mixture was refluxed (70 °C) for 12 hours. The progress of the reaction was monitored by TLC. After completion of the reaction, tert-butanol was evaporated in a rotary evaporator and the reaction mixture was partitioned between ethyl acetate and water. The collected organic layer was washed with water, aqueous ammonium chloride, brine solution and dried over anhydrous Na2SO4. After evaporation, the product was purified by column chromatography.
General Procedure for Deprotection of Tertiarybutyldimethylsilyl Ether: To a solution of respective TBDMS protected nucleoside (1eqv.) in THF, a solution of tetra-n- butylammoniumfluoride (TBAF) (2.5 eqv.) in THF was added. The reaction mixture was stirred at room temperature for 1 hour. After completion of the reaction, the solvent was evaporated in a rotary evaporator and was partitioned between ethyl acetate and water. The
collected organic layer was washed with brine solution and dried over anhydrous Na2SO4. After evaporation, the crude material obtained was purified by column chromatography.
Synthesis of 3',5'-di-O-tert-butyldimethylsilyl-5-(3-((4-(1-(4-bromophenyl)- triazolyl)phenyl)(methyl)amino)propynyl)-2'-deoxyuridine (2.44A):Using general procedure
for click reaction, starting from 100 mg (0.16 mmol) of compound 2.43 and 47.5 mg of 1- azido-4-bromobenzene (0.24 mmol), the title compound was isolated by Si-gel column chromatography (Hex: EtOAc = 3:1) as yellow solid. Yield: 52% (68 mg); m.p. 138-140 °C. IR (KBr): ̃ 3423, 2928, 2856, 1717, 1688, 1618 cm-1; 1H NMR (CDCl3, 600 MHz): 8.05 (s, 1H), 7.90 (s, 1H), 7.77 (d, J = 8.4 Hz, 2H), 6.90 (d, J = 8.4 Hz, 2H), 6.25 (t, J = 6.6 Hz, 2H), 4.36 (s,1H), 4.28 (s, 2H), 3.95 (s, 1H), 3.84 (d, J = 11.4 Hz, 1H), 3.72 (d, J = 11.4, 1H), 3.05 (s, 3H), 2.30-2.27 (m, 1H), 2.00-1.96 (m, 1H), 0.86 (s, 18H), 0.07 (s, 6H), 0.05 (s, 6H); 13C NMR (CDCl3, 150 MHz): 149.3, 149.1, 142.5, 136.3, 133.0, 122.2, 121.9, 119.7, 116.1, 114.1, 99.9, 90.1, 88.6, 86.1, 75.4, 72.6, 63.1, 43.3, 42.1, 38.7, 26.1, 25.9, -4.4, -4.6, -5.1, -5.4;
+ESI-HRMS calculated for C39H54BrN6O5Si2 [M+H]+ 821.2872, found 821.2873.
Synthesis of 3',5'-di-O-tert-butyldimethylsilyl-5-(3-((4-(1-(4-nitrophenyl)- triazolyl)phenyl)(methyl)amino)propynyl)-2'-deoxyuridine(2.44B):Using general procedure
for click reaction, starting from 100 mg (0.16 mmol) of compound 2.43 and 40 mg of 1- azido-4-nitrobenzene (0.24 mmol), the title compound was isolated by Si-gel column chromatography (Hex: EtOAc = 1:1) as rust brown solid. Yield: 52% (68 mg). IR (KBr): ̃ 3427, 2925, 1688, 1618, 1501 cm-1; 1H NMR (CDCl3, 600 MHz): s, 1H), 8.36 (d, J = 8.4 Hz, 2H), 8.18 (s, 1H), 7.97 (d, J = 8.4 Hz, 2H), 7.95 (s, 1H), 7.75 (d, J = 8.4 Hz, 2H), 6.88 (d, J = 8.4 Hz, 2H), 6.26 (t, J = 6 Hz, 1H), 4.37 (s,1H), 4.25 (s, 2H), 3.95 (s, 1H), 3.87 (d, J = 10.8 Hz, 1H), 3.74 (d, J = 11.4, 1H), 3.03 (s, 3H), 2.30-2.28 (m, 1H), 2.01-1.97 (m, 1H), 0.89 (s, 9H), 0.87 (s, 9H), 0.1 (s, 6H), 0.06 (s, 6H); 13C NMR (CDCl3, 150 MHz): 162.1, 149.6, 149.5, 149.4, 147.0, 142.6, 141.4, 127.1, 125.6, 120.2, 119.1, 116.0, 114.1, 99.9, 90.0, 88.6,
86.1, 75.7, 72.6, 63.1, 43.3, 42.2, 38.9, 26.2, 25.9, -4.5, -4.6, -5.2, -5.4; +ESI-HRMS calculated for C39H54N7O7Si2 [M+H]+ 788.3618, found 788.3617.
Synthesis of 3',5'-di-O-tert-butyldimethylsilyl-5-(3-((4-(1-(4-cyanophenyl)- triazolyl)phenyl)(methyl)amino)propynyl)-2'-deoxyuridine(2.44C):Using general procedure
for click reaction, starting from 100 mg (0.16 mmol) of compound 2.43 and 35 mg of 4- azidobenzonitrile (0.24 mmol), the title compound was isolated by Si-gel column chromatography (Hex: EtOAc = 2:1)as yellow solid. Yield: 94% (116 mg); m.p. 135-137 °C.
IR (KBr): ̃ 3415, 2929, 2857, 1699, 1619, 1607 cm-1; 1H NMR (CDCl3, 600 MHz): s, 1H), 8.36 (d, J = 8.4 Hz, 2H), 8.18 (s, 1H), 7.97 (d, J = 8.4 Hz, 2H), 7.95 (s, 1H), 7.75 (d, J = 8.4 Hz, 2H), 6.88 (d, J = 8.4 Hz, 2H), 6.26 (t, J = 6 Hz, 1H), 4.37 (s,1H), 4.25 (s, 2H), 3.95 (s, 1H), 3.87 (d, J = 10.8 Hz, 1H), 3.74 (d, J = 11.4, 1H), 3.03 (s, 3H), 2.30-2.28 (m, 1H), 2.01- 1.97 (m, 1H), 0.89 (s, 9H), 0.87 (s, 9H), 0.1 (s, 6H), 0.06 (s, 6H); 13C NMR (CDCl3, 150 MHz):
162.1, 149.6, 149.5, 149.4, 147.0, 142.6, 141.4, 127.1, 125.6, 120.2, 119.1, 116.0, 114.1, 99.9, 90.0, 88.6, 86.1, 75.7, 72.6, 63.1, 43.3, 42.2, 38.9, 26.2, 25.9, -4.5, -4.6, -5.2, -5.4; +ESI- HRMS calculated for C40H54N7O5Si2 [M+H]+ 768.3719, found 768.3717.
Synthesis of3',5'-di-O-tert-butyldimethylsilyl-5-(3-(methyl(4-(1-(4-(trifluoromethyl)-7- coumarinyl)-triazolyl)phenyl)amino)propynyl)-2'-deoxyuridine(2.44D): Using general
procedure for click reaction, starting from 100 mg (0.16 mmol) of compound 2.43 and 61 mg of 7-
azido-4-trifluoromethylcoumarin (0.24 mmol), the title compound was isolated by Si-gel column chromatography (Hex: EtOAc =1:1)as yellow solid. Yield: 94% (132 mg); m.p. 227-230 °C.
IR (KBr):̃3425, 2955, 2929, 1720, 1686, 1617, 1607 cm-1; 1H NMR (CDCl3, 600 MHz):
s, 1H), 8.17 (s, 1H), 7.92 (s, 1H), 7.85 (d, J = 12.6 Hz, 3H), 7.77 (d, J = 7.2 Hz, 2H), 6.89 (d, J = 6.6 Hz, 2H), 6.83 (s, 1H), 6.25 (t, J = 6.6 Hz, 2H), 4.37 (s,1H), 4.26 (s, 2H), 3.95 (s, 1H), 3.85 (d, J = 11.4 Hz, 1H), 3.73 (d, J = 10.8, 1H), 3.04 (s, 3H), 2.30-2.28 (m, 1H), 2.01- 1.98 (m, 1H), 0.88 (s, 9H), 0.87 (s, 9H), 0.08 (s, 6H), 0.06 (s, 6H); 13C NMR (CDCl3, 150 MHz): , 158.34, 155.3, 149.6, 149.5, 149.3, 142.5, 140.2, 127.1, 119.1, 116.4, 115.7,
114.1, 113.1, 108.5, 99.9, 89.9, 88.1, 75.6, 72.6, 63.16, 43.3, 42.2, 38.7, 29.8, 26.2, 25.9, 18.5, 18.2, -4.4, -4.6, -5.1, -5.4; +ESI-HRMS calculated for C43H54F3N6O7Si2[M+H]+ 879.3539, found 879.3531.
Synthesis of 3',5'-di-O-tert-butyldimethylsilyl-5-(3-((4-(1-(1-naphthyl)- triazolyl)phenyl)(methyl)amino)propynyl)-2'-deoxyuridine(2.44E): Using general procedure
for click reaction, starting from 104 mg (0.166 mmol) of compound 2.43 and 1-azidonaphthalene (42.25 mg, 0.25 mmol), the title compound was isolated by Si-gel column chromatography (Hex:
EtOAc = 2:1)as brown semi-solid. Yield: 53% (70 mg); m.p. 91-94 °C. IR (KBr): ̃ 3408, 2954, 2928, 2856, 1698, 1619 cm-1; 1H NMR (CDCl3, 400 MHz): 8.95s, 1H), 8.03 (s, 2H), 7.96 (bs, 1H), 7.90 (s, 1H), 7.83 (bs, 2H), 7.69 (bs, 1H), 7.59 (m, 5H), 6.93 (bs, 2H), 6.25 (s, 1H), 4.37 (s, 1H), 4.29 (s, 2H), 3.95 (s, 1H), 3.84 (bs, 1H), 3.74-3.73 (m, 1H), 3.06 (s, 3H), 2.28-2.28 (m, 1H), 1.99 (bs, 1H), 0.87 (s, 18H), 0.06 (s, 12H); 13C NMR (CDCl3, 100 MHz): 161.9, 149.3, 148.2, 142.5, 134.3, 130.4, 128.4, 127.2, 127.0, 125.2, 123.7, 122.7, 121.2, 120.0, 114.2, 99.9, 90.1, 88.6, 86.1, 77.5, 77.2, 76.9, 63.1, 43.3, 42.1, 38.7, 26.1, 25.9, -4.4, -4.6, -5.1, -5.4+ESI- HRMS calculated for C43H57N6O5Si2 [M+H]+ 793.3923, found 793.3925.
Synthesis of 3',5'-di-O-tert-butyldimethylsilyl-5-(3-((4-(1-(2-anthrayl)- triazolyl)phenyl)(methyl)amino)propynyl)-2'-deoxyuridine(2.44F):Using general procedure
for click reaction, starting from 100 mg (0.16 mmol) of compound 2.43 and 53 mg of 2-azidoanthracene (0.24 mmol), the title compound was isolated by Si-gel column chromatography (Hex: EtOAc = 3:1)as yellow solid. Yield: 57% (77 mg); m.p. 204- 208 °C. IR (KBr): ̃ 3406, 2953, 2928, 2856, 1699, 1619 cm-1; 1H NMR (CDCl3, 600 MHz):
δ 8.51 (s, 1H), 8.49 (s, 1H), 8.42 (s, 1H), 8.33 (s, 1H), 8.25 (s, 1H), 8.17 (d, J = 9.0 Hz, 1H), 8.03 (dd, J = 5.4, 3.6 Hz, 2H), 7.98 (dd, J = 9.0, 2.0 Hz, 1H), 7.91 (s, 1H), 7.84 (d, J = 8.4 Hz, 2H), 7.55 – 7.50 (m, 2H), 6.93 (d, J = 8.4 Hz, 2H), 6.25 (dd, J = 7.8, 6.0 Hz, 1H), 4.39 – 4.36 (m, 1H), 4.30 (s, 2H), 3.96 (d, J = 2.4 Hz, 1H), 3.85 (dd, J = 11.4, 2.4 Hz, 1H), 3.73 (dd, J = 11.4, 2.4 Hz, 1H), 3.07 (s, 3H), 2.31– 2.27 (m, 1H), 2.02 – 1.97 (m, 1H), 0.88 (s, 9H), 0.88 (s, 9H), 0.08 (d, J = 2.4 Hz, 6H), 0.06 (d, J = 4.8 Hz, 6H); 13C NMR (CDCl3, 150 MHz): δ 161.5,
149.3, 149.1, 142.4, 134.1, 132.6, 130.9, 130.7, 128.4, 128.2, 127.1, 127.0, 126.8, 126.4, 126.2, 119.9, 119.2, 117.8, 116.3, 114.1, 99.9, 88.5, 86.1, 77.1, 75.3, 72.6, 63.1, 43.3, 42.1, 38.7, 26.1, 25.8, 18.5, 18.1, -4.5, -4.7, -5.2, -5.46; +ESI-HRMS calculated for C47H59N6O5Si2 [M+H] + 843.4080, found 843.4085.
Synthesis of 3',5'-di-O-tert-butyldimethylsilyl-5-(3-((4-(1-(4-pyrenyl)- triazolyl)phenyl)(methyl)amino)propynyl)-2'-deoxyuridine (2.44G):Using general procedure
for click reaction, starting from 100 mg (0.16 mmol) of compound 2.43 and 58 mg of 1- azidopyrene (0.24 mmol), the title compound was isolated by Si-gel column chromatography (Hex: EtOAc = 3:1) as pale yellow solid. Yield:
77% (107 mg); m.p. 135-138 °C. IR (KBr): ̃ 3423, 3046, 2954, 2928, 2856, 1699, 1619 cm-1; 1H NMR (CDCl3, 600 MHz): δ 8.29 (dd, J = 6.0, 3 Hz, 2H), 8.26 (d, J = 6.6 Hz, 1H), 8.20 (d, J = 9.0 Hz, 1H), 8.16 (s, 1H), 8.16 (s, 1H), 8.14 (s, 1H), 8.13 – 8.08 (m, 2H), 7.96 (d, J = 9.0 Hz, 1H), 7.90 (d, J = 6.0 Hz, 2H), 7.88 (s, 1H), 6.96 (d, J = 9.0 Hz, 2H), 6.25 (dd, J = 7.8, 5.4 Hz, 1H), 4.38 (dt, J = 5.4, 2.4 Hz, 1H), 4.31 (s, 2H), 3.96 (q, J = 2.4 Hz, 1H), 3.86 (dd, J = 11.4, 2.4 Hz, 1H), 3.74 (dd, J = 11.4, 1.8 Hz, 1H), 3.08 (s, 3H), 2.32 – 2.27 (m, 1H), 2.03 – 1.97 (m, 1H), 0.90 (s, 9H), 0.88 (s, 9H), 0.09 (d, J = 3.0 Hz, 6H), 0.07 (d, J = 4.8 Hz, 6H); 13C NMR (CDCl3, 150 MHz): δ 161.4, 149.3, 149.1, 148.3, 142.4, 132.3, 131.3, 130.8, 130.8, 129.7, 129.0, 127.2, 127.1, 126.9, 126.5, 126.38, 126.2, 125.2, 124.9, 124.36, 123.5, 121.6, 121.5, 120.1, 114.2, 99.9, 90.1, 88.6, 86.1, 77.4, 77.1, 76.9, 75.3, 72.6, 63.1, 43.3, 42.1, 38.7, 26.1, 25.8, 18.5, 18.1, -4.5, -4.7, -5.2, -5.4; +ESI- HRMS calculated for C49H59N6O5Si2 [M+H] + 867.4080, found 867.4082.
Synthesis of 5-(3-((4-(1-(4-cyanophenyl)-triazolyl)phenyl)(methyl)amino)propynyl)-2'- deoxyuridine (2.45C):Using general procedure for TBDMS deprotection, starting from 230 mg (0.3 mmol) of nucleoside 2.44C, the title compound was isolated by Si-gel column chromatography (CHCl3: MeOH = 10:1) as orange-yellow solid. Yield: 81%
(130 mg); m.p. 178-181 °C. IR (KBr): ̃ 3426, 3056, 2925, 2837, 2229, 1714, 1607, 1619 cm-
1; 1H NMR (DMSO-d6, 600 MHz): δ 11.54 (s, 1H), 9.23 (s, 1H), 8.15 – 8.12 (m, 3H), 8.09 (d, J = 9.0 Hz, 2H), 7.74 (d, J = 9.0 Hz, 2H), 6.95 (d, J = 9.0 Hz, 2H), 6.04 (t, J = 6.6 Hz, 1H), 5.21 (d, J = 4.2 Hz, 1H), 5.06 (t, J = 5.0 Hz, 1H), 4.34 (s, 2H), 4.20 – 4.15 (m, 1H), 3.74 (q, J
= 3.6 Hz, 1H), 3.59 – 3.53 (m, 1H), 3.54 – 3.49 (m, 1H), 2.95 (s, 3H); 13C NMR (DMSO-d6, 150 MHz): δ 161.5, 149.4, 149.1, 148.3, 143.7, 139.6, 134.3, 126.4, 120.2, 118.8, 118.2, 117.9, 113.9, 110.8, 98.1, 88.6, 87.6, 84.8, 76.1, 70.1, 60.9, 42.1, 40.2, 40.1, 38.0; +ESI-HRMS calculated for C28H26N7O5[M+H]+ 540.1990, found 540.1990.
Synthesis of 5-(3-((4-(1-(naphthayl)-triazolyl)phenyl)(methyl)amino)propynyl)-2'- deoxyuridine(2.45E): Using general procedure for TBDMS deprotection, starting from 94 mg
(0.12 mmol) of 2.44E, the title compound was isolated by Si-gel column chromatography (CHCl3: MeOH = 20:1)as pale yellow solid. Yield: 80% (53 mg); m.p.
152-155 °C. IR (KBr): ̃ 3417, 3059, 2922, 2851, 1691, 1618cm-1; 1H NMR (600 MHz, DMSO-d6): δ 11.61 (s, 1H), 8.96 (s, 1H), 8.21 (d, J = 8.4 Hz, 1H), 8.18 (s, 1H), 8.14 (d, J = 7.8 Hz, 1H), 7.83 (d, J = 8.7 Hz, 2H), 7.80 (d, J = 7.0 Hz, 1H), 7.75 – 7.70 (m, 1H), 7.70 – 7.59 (m, 3H), 6.99 (d, J = 9.0 Hz, 2H), 6.08 (t, J = 6.6 Hz, 1H), 5.25 (d, J = 4.2 Hz, 1H), 5.12 (t, J = 5.0 Hz, 1H), 4.39 (s, 2H), 4.22 (p, J = 4.2 Hz, 1H), 3.78 (q, J = 3.2 Hz, 1H), 3.63 – 3.58 (m, 1H), 3.58 – 3.53 (m, 1H), 2.99 (s, 3H); 13C NMR (DMSO-d6, 150 MHz): δ 172.2, 161.6, 149.5, 148.9, 147.1, 143.7, 133.7, 133.5, 130.2, 128.4, 128.1, 127.9, 127.2, 126.4, 125.5, 123.8, 122.6, 122.2, 119.5, 114.1, 98.2, 88.7, 87.6, 84.8, 79.2, 76.2, 70.1, 60.9, 45.7, 42.3, 40.2, 40.1, 38.1; +ESI-HRMS calculated for C31H29N6O5 [M+H]+ 565.2194 , found 565.2203.
Synthesis of 5-(3-((4-(1-(pyrenyl)triazolyl)phenyl)(methyl)amino)propynyl)-2'- deoxyuridine (2.45G):Using general procedure for TBDMS deprotection, starting from 158 mg (0.18 mmol) of 2.44G, the title compound was isolated by Si-gel column chromatography (CHCl3: MeOH = 20:1) as light brown solid. Yield: 85% (99 mg); m.p.
217-220 °C. IR (KBr): ̃ 3420, 3054, 2924, 2851, 1691, 1618cm-1; 1H NMR (DMSO-d6, 600 MHz): δ 11.60 (s, 1H), 9.10 (s, 1H), 8.52 (d, J = 8.4 Hz, 1H), 8.46 (d, J = 7.2 Hz, 1H), 8.42 (d, J = 7.8 Hz, 1H), 8.40 – 8.33 (m, 3H), 8.30 (d, J
= 9.0 Hz, 1H), 8.22 – 8.17 (m, 2H), 7.93 – 7.87 (m, 3H), 7.01 (d, J = 8.4 Hz, 2H), 6.09 (t, J = 6.6 Hz, 1H), 5.25 (d, J = 4.2 Hz, 1H), 5.12 (t, J = 4.8 Hz, 1H), 4.40 (s, 2H), 4.22 (p, J = 4.2 Hz, 1H), 3.79 (q, J = 3.6 Hz, 1H), 3.64 – 3.59 (m, 1H), 3.59 – 3.54 (m, 1H), 3.01 (s, 3H); 13C NMR (DMSO-d6, 150 MHz): δ 161.6, 149.5, 148.9, 147.3, 143.7, 131.7, 130.7, 130.5, 130.2, 129.7, 128.86, 127.2, 127.2, 126.6, 126.4, 126.2, 125.4, 125.2, 124.1, 123.8, 123.4, 123.0, 121.2,
119.5, 114.1, 98.2, 88.7, 87.6, 84.8, 79.2, 76.2, 70.1, 60.9, 42.3, 40.1, 40.1, 39.5, 38.1; +ESI- HRMS calculated for C37H31N6O5 [M+H]+ 639.2350, found 639.2313.