SUPPORTING INFORMATION
The (5Z)-5-Pentacosenoic and 5-Pentacosynoic Acids Inhibitthe
HIV-1 Reverse Transcriptase
Lizabeth Giménez Moreira, Elsie A. Orellano, Karolyna Rosado, Rafael V. C. Guido, Adriano D. Andricopulo, Gabriela Ortiz Soto, José W. Rodríguez, David J. Sanabria-Ríos, Néstor M. Carballeira*
Table of Contents
InstrumentationS2
Synthesis of 2-(4-bromobutoxy)-tetrahydro-2H-pyran (4a)S2
Synthesis of 2-(8-bromooctoxy)-tetrahydro-2H-pyran (4b)S3
Synthesis of trimethyl [6-(tetrahydro-2H-pyran-2-yloxy)-1-hexynyl] silane (5a)S3
Syntheis of trimethyl [10-(tetrahydro-2H-pyran-2-yloxy)-1-decynyl] silane (5b)S3
Synthesis of 2-[(5-hexynyl)oxy]-tetrahydro-2H-pyran (6a)S4
Synthesis of 2-[(9-decynyl)oxy]-tetrahydro-2H-pyran (6b)S4
Synthesis of 2-[(5-pentacosynyl)oxy]-tetrahydro-2H-pyran (7a)S4
Synthesis of 2-[(9-pentacosynyl)oxy]-tetrahydro-2H-pyran (7b)S5
Synthesis of 5-pentacosyn-1-ol (8a)S6
Synthesis of 9-pentacosyn-1-ol (8b)S6
Synthesis of 5-pentacosynoic acid (1a)S7
Synthesis of 9-pentacosynoic acid (1b)S8
Synthesis of (5Z)-5-pentacosen-1-ol (9a)S8
Synthesis of (9Z)-pentacosen-1-ol (9b)S9
Synthesis of (5Z)-5-pentacosenoic acid (2a)S10
Synthesis of (9Z)-9-pentacosenoic acid (2b)S10
Synthesis of 2-pentacosynoic acidS11
ReferencesS11
Instrumentation
All compounds were analyzed by 1H NMR (300 or 500 MHz) and 13C NMR (75 or 125 MHz) by using a Bruker Avance DPX-300 or a Bruker Avance DRX-500 spectrometer. The samples were diluted in 99.8% chloroform-d (CDCl3) and the solvent signals at 7.26 and 77.0 ppm were used as internal standard for proton and carbon, respectively. Mass spectral data was acquired on a GC-MS (Hewlett-Packard 5972A MS ChemStation or Agilent 5975C MS Chemstation) instrument at 70 eV, equipped with a 30 x 0.25 mm special performance capillary column (HP-5MS) of polymethylsiloxane cross-linked with 5% phenyl methylpolysiloxane. Infrared spectra were recorded on a Nicolet Magna 750 FT-IR spectrophotometer (Thermo-Nicolet, Madison, WI, USA).
2-(4-Bromobutoxy)-tetrahydro-2H-pyran (4a)
Into a round-bottomed flask equipped with a magnetic stirrer was added a solution of 4-bromo-1-butanol (1.29 g, 8.4 mmol) in 20 mL of CHCl3. To this solution, the 3,4-dihydro-2H-pyran (1.77 g, 21.1 mmol) was added drop wise followed by the addition of catalytic amounts of p-toluenesulfonic acid (p-TSA). The reaction mixture was stirred for 3h at rt. The chloroform was evaporated in vacuo and the organic layer was washed with an aqueous NaHCO3 saturated solution (3 x 30 mL), dried over MgSO4, filtered and concentrated in vacuo. The crude product was initially purified under Kugel Rohr distillation (95-120 °C/3 mm Hg) for 3 h and then further purified using silica gel column chromatography with hexane/ether (9:1 v/v) as the mobile phase affording 4a as a colorless oil in 99% yield with spectral data comparable to those previously reported in the literature [1].
2-(8-Bromooctoxy)-tetrahydro-2H-pyran (4b)
The tetrahydropyran4b was obtained as colorless oil in 93% yield from the reaction of 8-bromo-1-octanol (1.98 g, 6.8 mmol) with 3,4-dihydro-2H-pyran (0.7 g, 8.2 mmol) and catalytic amounts of p-TSA in CHCl3 (20 mL) following the procedure described above and with spectral data similar to the one reported in the literature [1].
Trimethyl [6-(tetrahydro-2H-pyran-2-yloxy)-1-hexynyl] silane (5a)
To a stirred solution of (trimethylsilyl)acetylene (1.94 g, 19.7 mmol) in dry THF (10.0 mL) and under a nitrogen atmosphere, n-BuLi (2.5 M, 10.0-26.0 mmol) in hexane was added drop wise at -78.0 °C. After 5 min HMPA (3.0 mL) was added followed by the drop wise addition of 4a (1.0 g, 6.6 mmol) in THF (10.0 mL) while maintaining the temperature at -78 °C. After 24 h, the reaction mixture was quenched with water, extracted with diethyl ether (2 x 15 mL) and washed with brine (2 x 20 mL). The organic phase was dried with MgSO4, filtered, and the solvent rotoevaporated. Final purification of the product by Kugel Rohr distillation (110-120 °C/3 mm Hg) afforded 5a as a colorless oil in a 92% yield with spectral data comparable to those previously reported in the literature [2].
Trimethyl [10-(tetrahydro-2H-pyran-2-yloxy)-1-decynyl] silane (5b)
Silane5b was obtained as a colorless oil [3] in 94% yield from the reaction of 1.70 mL of (trimethylsilyl)acetylene (1.21 g, 12.3 mmol) and 4b (1.20 g, 4.1 mmol) following the procedure described for 5a.
2-[(5-Hexynyl)oxy]-tetrahydro-2H-pyran (6a)
A mixture of 5a (0.99 g, 3.9 mmol) in 20.0 mL of dry THF was stirred at 0 °C and then tetrabutylammonium fluoride (2.3 mL, 7.8 mmol) was added drop-wise to the stirred solution. After 2 h, the reaction mixture was quenched with a 2M HCl solution and extracted with diethyl ether (2 x 20 mL). The organic extracts were dried over MgSO4 and concentrated in vacuo. The product was purified by Kugel Rohr distillation (110-120 °C/3 mm Hg), affording 6a in a 99% yield as a colorless oil and with spectral data similar to the one reported in the literature [4].
2-[(9-Decynyl)oxy]-tetrahydro-2H-pyran (6b)
The tetrahydropyran6b was obtained as a colorless oil [3] and in a 98% yield from the reaction of 1.66 g (5.3 mmol) of 5b and 3.1 mL (10.7 mmol) of TBAF following the procedure described for 6a.
2-[(5-Pentacosynyl)oxy]-tetrahydro-2H-pyran (7a)
To a solution of 6a (0.19 g, 1.0 mmol) and dry THF (15 mL) at -78°C, n-BuLi (2.5 M, 2.1-3.2 mmol) in hexane was added drop-wise while stirring under an argon atmosphere. After 45 min, HMPA (3.0 mL) was added followed by the addition of 1-bromononadecane (0.32 g, 0.9 mmol). The reaction was left stirring for 24 h at room temperature. The reaction mixture was then quenched with water and the organic products extracted with diethyl ether (1 x 50 mL), dried over MgSO4, filtered and evaporated in vacuo. The crude products were initially purified under Kugel Rohr distillation (160-195 °C/3 mm Hg) for 4-5 h and then further purified using alumina column chromatography with hexane first and then diethyl ether as the mobile phases affording 0.34 g of 7a in an overall 81% yield. The product was used as such for the next step without further purification, mp 61-63°C, IR (neat) νmax 2919, 2851, 1726, 1464, 1119, 1023, 811, 721, 626 cm-1; 1H NMR (CDCl3, 300 MHz) δ 4.54-4.53 (1H, t, J = 2.7 Hz), 3.85-3.68 (2H, m), 3.48-3.33 (2H, m), 2.19-2.07 (4H, m, H-4, H-7), 1.84-1.61 (6H, m), 1.59-1.48 (6H, m), 1.45-1.22 (32H, m, -CH2-), 0.86-0.82 (3H, t, J = 7.2 Hz, H-25); 13C NMR (CDCl3, 75 MHz) δ 98.7 (d), 80.4 (s), 79.7 (s), 67.0 (t, C-1), 62.1 (t), 31.9 (t, C-23), 30.7 (t), 29.7 (t), 29.6 (t), 29.5 (t), 29.4 (t), 29.3 (t), 29.2 (t), 29.1 (t), 28.9 (t), 28.7 (t), 28.1 (t), 25.9 (t), 25.5 (t, C-3), 22.7 (t, C-24), 19.7 (t), 19.6 (t), 18.7 (t), 18.6 (t), 14.1 (q, C-25).
2-[(9-Pentacosynyl)oxy]-tetrahydro-2H-pyran (7b)
The tetrahydropyran7b was obtained in a 60% yield as a white solid from the reaction of 0.20 g (0.8 mmol) of 6b and 1-bromopentadecane (0.22 g, 0.8 mmol) following the procedure described for 7a. The product was used as such for the next step without further purification, mp 57-59°C, IR (neat) νmax 2915, 2848, 1462, 1122, 1027, 719 cm-1; 1H NMR (CDCl3, 300 MHz) δ 4.58-4.56 (1H, t, J = 2.7 Hz), 3.90-3.69 (2H, m), 3.53-3.33 (2H, m), 2.20-2.10 (4H, m, H-8, H-11), 1.94-1.65 (6H, m), 1.62-1.42 (6H, m), 1.31-1.25 (32H, m, -CH2-), 0.89-0.85 (3H, t, J = 6.9 Hz, H-25); 13C NMR (CDCl3, 75 MHz) δ 98.8 (d), 84.8 (s), 80.3 (s), 67.7 (t, C-1), 62.3 (t), 31.9 (t, C-23), 30.8 (t, C-2), 29.69 (t), 29.65 (t), 29.6 (t), 29.4 (t), 29.35 (t), 29.2 (t), 29.1 (t), 28.9 (t), 28.8 (t), 28.76 (t), 28.7 (t), 28.4 (t), 28.2 (t), 26.2 (t), 25.5 (t), 22.7 (t, C-24), 19.7 (t), 18.7 (t), 18.4 (t), 14.1 (q, C-25).
5-Pentacosyn-1-ol (8a)
Pyran7a (0.34 g, 0.8 mmol) and catalytic amounts of p-TSA in methanol (15.0 mL) were stirred at 45 °C for 3 h. The reaction mixture was quenched with a 5% NaHCO3 solution and extracted with diethyl ether (2 x 20 mL). The organic extracts were dried over MgSO4 and concentrated in vacuo. The product was purified by Kugel Rohr distillation (160-190°C/ 3 mm Hg), affording 8a in a 99% yield as a white solid, mp 57-59 °C, IR (neat) νmax 3455 (br, -OH), 2921, 2852, 2179, 1460, 1377, 1258, 1121, 1076, 1023, 798, 721 cm-1; 1H NMR (CDCl3, 300 MHz) δ 3.69-3.65 (2H, t, J = 6.3 Hz, H-1), 2.22-2.10 (4H, m, H-4, H-7), 1.72-1.63 (2H, m, H-2), 1.61-1.51 (4H, m, H-3, H-8), 1.49-1.42 (2H, br. quintet, J = 7.1 Hz, H-24), 1.34-1.25 (31H, m, -CH2-, -OH), 0.89-0.85 (3H, t, J = 6.9 Hz, H-25); 13C NMR (CDCl3, 75 MHz) δ 80.8 (s), 79.7 (s), 62.5 (t, C-1), 31.9 (t, C-2), 31.85 (t, C-23), 29.7 (t), 29.65 (t), 29.55 (t), 29.4 (t), 29.2 (t), 29.1 (t), 28.9 (t), 25.3 (t, C-3), 22.7 (t, C-24), 18.7 (t), 18.5 (t), 14.1 (q, C-25). HRMS (APCI) Calcd for C25H49O1 [M + H]+ 365.3778, found 365.3778.
9-Pentacosyn-1-ol (8b)
The 9-pentacosyn-1-ol (8b) was obtained in an 83% yield as a white solid from the reaction of 0.21 g (0.5 mmol) of 7b in methanol with catalytic amounts of p-TSA according to the procedure described for 8a, mp 54-55°C, IR (neat) νmax 3265 (br, -OH), 2915, 2848, 2116, 1485, 1471, 1462, 1405, 1353, 1122, 1061, 1031, 1014, 729, 719, 682 cm-1; 1H NMR (CDCl3, 300 MHz) δ 3.65-3.61 (2H, t, J = 6.7 Hz, H-1), 2.15-2.10 (4H, t, J = 6.7 Hz, H-8, H-11), 1.58-1.51 (2H, m, H-2), 1.46-1.44 (4H, quintet, J = 6.9 Hz, H-7, H-12), 1.38-1.24 (33H, m, –CH2-, -OH), 0.89-0.85 (3H, t, J = 6.9 Hz, H-25); 13C NMR (CDCl3, 75 MHz) δ 80.3 (s), 80.1 (s), 63.0 (t, C-1), 32.7 (C-2), 31.9 (t, C-23), 29.7 (t), 29.6 (t), 29.55 (t), 29.34 (t), 29.30 (t), 29.2 (t), 29.1 (t), 28.9 (t), 28.8 (t), 25.7 (t, C-3), 22.7 (t, C-24), 18.7 (t), 14.1 (q, C-25). HRMS (APCI) Calcd for C25H49O1 [M + H]+ 365.3778, found 365.3778.
5-Pentacosynoic acid (1a)
To a stirred solution of 8a (0.10 g, 0.3 mmol) in 10.0 mL of DMF was slowly added a solution of pyridinium dichromate (0.42 g, 1.1 mmol) at rt. After 24 h, the reaction mixture was washed with water (3 x 25 mL), diethyl ether (1 x 25 mL), dried over MgSO4, filtered, and concentrated in vacuo. The crude product was purified utilizing fluorisil column chromatography eluting with diethyl ether affording 1a as a white solid in a 59% yield, mp 64-66°C, IR (neat) νmax 3151-3091, 2916, 2849, 1705 (C=O), 1463, 1378, 1295, 1186, 1099, 937, 721 cm-1; 1H NMR (CDCl3, 500 MHz) δ 2.51-2.48 (2H, t, J = 7.4 Hz, H-2), 2.26-2.23 (2H, t, J = 6.8 Hz, H-7), 2.14-2.13 (2H, t, J = 7.2 Hz, H-4), 1.84-1.78 (2H, br. quintet, J = 6.9 Hz, H-3), 1.50-1.44 (2H, br. quintet, J = 7.3 Hz, H-8), 1.35-1.25 (32H, m, -CH2-), 0.89-0.87 (3H, t, J = 6.9 Hz, H-25); 13C NMR (CDCl3, 75 MHz) δ 179.5 (s, C-1), 81.5 (s, C-6), 78.5 (s, C-5), 32.8 (t, C-2), 31.9 (t, C-23), 29.7 (t), 29.5 (t), 29.4 (t), 29.2 (t), 29.0 (t), 28.9 (t), 23.9 (t, C-3), 22.7 (t, C-24), 18.7 (t, C-7), 18.1 (t, C-4), 14.1 (q, C-25). HRMS (APCI) Calcd for C25H47O2 [M + H]+ 379.3570, found 379.3571.
9-Pentacosynoic acid (1b)
The 9-pentacosynoic acid (1b) was obtained in an 80% yield as a white solid from the reaction of 0.019 g (0.05 mmol) of 8b and 0.12 g (0.3 mmol) of PDC following the procedure described for 1a, mp 61-63°C, IR (neat) νmax 3091 (br, -OH), 2916, 2849, 1705 (C=O), 1463, 1432, 1412, 1378, 1295, 1099, 937, 721 cm-1; 1H NMR (CDCl3, 500 MHz) δ 2.36-2.32 (2H, t, J = 7.5 Hz, H-2), 2.15-2.09 (4H, m, H-8, H-11), 1.63-1.50 (2H, quintet, J = 6.6 Hz, H-3), 1.48-1.37 (4H, quintet, J = 6.9 Hz, H-7, H-12), 1.31-1.25 (30H, m, -CH2-) 0.89-0.86 (3H, t, J = 6.9 Hz, H-25); 13C NMR (CDCl3, 125 MHz) δ 179.6 (s, C-1), 80.3 (s, C-10), 80.0 (s, C-9), 34.0 (t, C-2), 31.9 (t), 29.71 (t), 29.67 (t), 29.6 (t), 29.44 (t), 29.36 (t), 28.9 (t), 28.8 (t), 28.5 (t), 24.7 (t), 22.7 (t), 18.75 (t), 18.71 (t), 14.1 (q, C-25).
(5Z)-5-Pentacosen-1-ol (9a)
Into a 25-mL two-necked round-bottomed flask were placed dry hexane, 8a (72 mg, 0.2 mmol), quinoline (0.3 mL), and palladium in activated carbon (Lindlar’s catalyst). One of the necks was capped with a rubber septum and the other was connected via tygon tubing to a 25-mL graduated pipet ending in a 150-mL beaker with distilled water. While stirring at rt a 20-mL syringe with needle was used to withdraw air from the system and to draw water up the graduated pipet to the 0.0 mL mark. Hydrogen was then introduced into the system using a balloon filled with hydrogen attached to a hose barb-to-luer lock adapter with a stopcock and a needle. The reaction mixture consumed 4.8-9.3 mL of hydrogen during 1 h. The mixture was filtered and the solvent removed in vacuo. The pentacosenol9a was purified under Kugel Rohr distillation (160-190 °C/ 3 mm Hg), affording a white solid, in a 97% yield. The alkenol was used as such for the next step without further purification, mp 50-52 °C, IR (neat) νmax 3346 (OH, broad), 2916, 2849, 1621, 1468, 1352, 1259, 1200, 1121, 1064, 1023, 978, 905, 868, 810, 718 cm-1; 1H NMR (CDCl3, 500 MHz) δ 5.40-5.31 (2H, m, H-5, H-6), 3.66-3.63 (2H, t, J = 6.5 Hz, H-1), 2.08-1.97 (5H, m, H-4, H-7, -OH), 1.61-1.56 (2H, br. quintet, J = 7.1 Hz, H-2), 1.45-1.39 (2H, br. quintet, J = 7.5 Hz, H-3), 1.32-1.25 (34H, m, -CH2-), 0.89-0.86 (3H, t, J = 6.4 Hz, H-25); 13C NMR (CDCl3, 125 MHz) δ 130.4 (d), 129.3 (d), 62.9 (t, C-1), 32.4 (t, C-2), 32.3 (t, C-23), 31.9 (t), 31.74 (t), 29.69 (t), 29.65 (t), 29.6 (t), 29.4 (t), 29.3 (t), 29.2 (t), 27.3 (t), 26.9 (t), 25.8 (t, C-3), 22.7 (t, C-24), 14.1 (q, C-25).
(9Z)-Pentacosen-1-ol (9b)
The pentacosenol9b was obtained in a 70 % yield as a white solid from the catalytic hydrogenation, using Lindlar’s catalyst, of 8b (0.14 g, 0.4 mmol) following the procedure described for 9a, mp 47-49°C, IR (neat) νmax 3311-2900 (br, -OH), 2920, 2851, 1641, 1463, 1259, 1083, 1013, 867, 793 cm-1; 1H NMR (CDCl3, 300 MHz) δ 5.35-5.32 (2H, m, H-9, H-10), 3.66-3.61 (2H, t, J = 6.7 Hz, H-1), 2.13-2.11(4H, m, H-8, H-11), 1.56-1.18 (38H, m, -CH2-), 0.89-0.85 (3H, t, J = 6.7 Hz, H-25); 13C NMR (CDCl3, 75 MHz) δ 130.0 (d, C-10), 129.8 (d, C-9), 63.1 (t, C-1), 32.8 (t, C-2), 31.9 (t, C-23), 29.7 (t), 29.6 (t), 29.5 (t), 29.4 (t), 29.2 (t), 28.9 (t), 28.8 (t), 28.1 (t), 27.2 (t), 25.7 (t, C-3), 22.7 (t, C-24), 14.1 (q, C-25).
(5Z)-5-Pentacosenoic acid (2a)
Was obtained in 83% yield as a white solid from the reaction of 0.070 g (0.2 mmol) of 9a and PDC (0.43 g, 1.1 mmol) using the procedure described for 1a, mp 56-58°C, IR (neat) νmax 3091(br, -OH), 2915, 2848, 1693 (C=O), 1464, 1412, 1294, 1225, 1206, 1187, 1099, 1039, 1026, 932, 720, 688 cm-1; 1H NMR (CDCl3, 500 MHz) δ 5.46-5.29 (2H, m, H-5, H-6), 2.43-2.33 (2H, m, H-2), 2.12-1.96 (4H, m, H-4, H-7), 1.73-1.63 (2H, m, H-3), 1.32-1.25 (36H, m, -CH2-) 0.89-0.86 (3H, t, J = 6.8 Hz, H-25); 13C NMR (CDCl3, 125 MHz) δ 179.8 (s, C-1), 131.4 (d, C-6), 128.1 (d, C-5), 33.3 (t, C-2), 31.9 (t, C-23), 29.7 (t), 29.6 (t), 29.4 (t), 27.2 (t, C-7), 26.4 (t, C-4), 24.6 (t, C-3), 22.7 (t, C-24), 14.1 (q, C-25).
(9Z)-9-Pentacosenoic acid (2b)
Acid 2b was obtained in 66% yield as a white solid from the reaction of 0.098 g (0.3 mmol) of 9b and 0.60 g (1.6 mmol) of PDC using the procedure described for 1a, mp 54-56°C, IR (neat) νmax 3084 (br, -OH), 2915, 2848, 1702 (C=O), 1463, 1411, 1296, 1099, 1026, 937, 721 cm-1; 1H NMR (CDCl3, 300 MHz) δ 5.34-5.33 (2H, m, H-9, H-10), 2.36-2.30 (2H, t, J = 7.4 Hz, H-2), 2.01-1.99 (4H, m, H-8, H-11), 1.62-1.60 (2H, m, H-3), 1.29-1.24 (37H, m, -CH2-) 0.88-0.85 (3H, t, J = 6.9 Hz, H-25); 13C NMR (CDCl3, 75 MHz) δ 180.0 (s, C-1), 130.0 (d, C-10), 129.7 (d, C-9), 34.0 (t, C-2), 31.9 (t, C-23), 29.7 (t), 29.64 (t), 29.59 (t), 29.53 (t), 29.4 (t), 29.35 (t), 29.3 (t), 29.23 (t), 29.19 (t), 29.12 (t), 29.07 (t), 28.97 (t), 27.2 (t), 27.1 (t), 24.7 (t, C-3), 22.7 (t, C-24), 14.1 (q, C-25).
2-Pentacosynoic acid
The 2-pentacosynoic acid was obtained in a 13% yield as a white solid from the reaction of 1-tetracosyne (0.035 g, 1.0 mmol) and excess dry CO2 following the standard procedure, mp 69-70°C, IR (neat) νmax 3456-3045 (br), 2956, 2915, 2850, 2251, 1723, 1467, 1379, 1269, 1072, 742, 714 cm-1; 1H NMR (CDCl3, 300 MHz) δ 2.37-2.32 (2H, t, J = 7.2 Hz, H-4), 1.63-1.54 (2H, quintet, J = 7.4 Hz, H-5), 1.41-1.26 (38H, m, -CH2-), 0.89-0.85 (3H, t, J = 7.2 Hz, H-25); 13C NMR (CDCl3, 125 MHz) δ 158.2 (s, C-1), 92.7 (s, C-3), 72.6 (s, C-2), 31.9 (t, C-23), 29.64 (t), 29.62 (t), 29.55 (t), 29.4 (t), 29.3 (t), 29.0 (t), 28.8 (t), 27.4 (t), 22.7 (t, C-24), 18.8 (t, C-4), 14.1 (q, C-25).
References
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