Resultados y Discusiones 53 0.8 Hz, H-16); 1.56 (m, H-1a); 1.45-1.30 (5H, m, H2-2, H2-3, H-9 ); 1.18 (s, H-17); 1.16 (s, H-18);
1.15 (d,J= 10.8 Hz, H-5 ); 1.05-0.92 (m, H-1b); 1.01 (s, H-20); 0.90 (s, H-19)
RMN13C (CDCl3, 75 MHz); 133.55 (C-14), 133.29 (C-12), 130.99 (C-13), 113.88 (C-15), 84.99 (C-7), 77.00 (C-8), 71.70 (C-6), 59.57 (C-9), 57.19 (C-5), 43.32 (C-3), 39.84 (C-1), 39.14 (C-10), 36.23 (C-18), 33.74 (C-4), 22.72 (C-11), 22.15 (C-20), 19.87 (C-16), 19.38 (C-17), 18.19 (C-2), 16.93 (C-19)
EM-IE+:m/z(% intensidad relativa) 322 (no observado), 304 (9, [M H2O]+), 286 (8, 304 H2O), 81 (94), 69 (90), 55 (73), 43 (100), 41 (82), 29 (19)
OH
OH OH
H
15 16
9 5 7 3
1
20 17
19 18 11
13
117
6.2.3 6-O-Acetilaustroinulina (118) [34, 35]
S lido blanco y cristales transparentes (5.541g) Rf 0.50 (Hex:AcOEt, 55:45)
Pf 162-165 C
[ ]D25+ 34.2 (c= 1.0, CHCl3)
IR CHCl3 max3581 (OH); 2932, 2875; 1726 (C=O); 1253 (C-O); 1641, 975 y 909cm-1(C=C) RMN1H (CDCl3, 300 MHz); 6.87 (ddd,J= 0.9, 10.8, 17.1 Hz, H-14); 5.44 (t,J= 7.2 Hz, H-12);
5.19 (d,J = 17.4 Hz, H-15); 5.12 (dd, J= 9.9, 11.4 Hz, H-6 ); 5.10 (dd,J = 11.4, 2.1 Hz, H-15);
3.46 (d,J= 10.2 Hz, H-7 ); 2.48 (ddd,J= 1.2, 4.5, 15.9 Hz, H-11a); 2.25 (dt,J= 6.0, 16.2 Hz, H- 11b); 2.12 (s, H3C-Ac); 1.79 (d, J = 1.2 Hz, H3-16); 1.61 (ddt, J = 13.0, 5.0, 1.5 Hz, H-1a); 1.56 (ddt, J= 13.5, 13.0, 4.0 Hz, H-2a); 1.41 (dt,J = 14.5, 5.0 Hz, H-2b); 1.40 (d, J= 11.0 Hz, H-5 );
1.39 (t,J= 5.5 Hz, H-9 ); 1.34 (dtd,J= 113.5, 5.0, 1.5 Hz, H-3a); 1.23 (s, H-17); 1.22 (m, H-3b);
1.01 (s, H-18); 0.96 (s, H-20); 0.95-0.97 (m, H-1b); 0.87 (s, H-19)
RMN 13C (CDCl3, 75 MHz); , asignaci n: 171.81 (C=O-Ac), 133.61 (C-12), 133.16 (C-14), 130.99 (C-13), 113.79 (C-15), 83.67 (C-7), 76.89 (C-8), 73.19 (C-6), 59.15 (C-9), 56.49 (C-5), 43.39 (C-3), 39.81 (C-1), 39.39 (C-10), 35.99 (C-18), 33.33 (C-4), 22.75 (C-11), 22.17 (C-19), 21.79 (CH3-Ac), 19.85 (C-16), 19.54 (C-17), 18.03 (C-2), 16.75 (C-20)
EM-IE+: m/z (% intensidad relativa) 364 (no observado), 346 (10, [M H2O]+), 286 (65, 346 C2H4O2), 205 (95), 137 (67), 81 (93), 43 (100), 18 (10)
EM-FAB+:m/z(% intensidad relativa) 364 (1,M+), 347 (17, [M H2O]+), 287 (55, 347 C2H4O2), 205 (25), 81 (100), 69 (38), 43 (35), 29 (7)
EM-AR-FAB+: Observado:m/z347.2589; Calculado: 347.2586 [M H2O + H]+).
OH
OH O
H
15 16
9 5 7 3
1
20 17
19 18 11
13
O
118
6.2.4 Derivati aci n qu mica de 6-O-acetilaustroinulina (118)
6.2.5 6,7-Diacetilaustroinulina (118a): 62.5 mg de 118 fueron disueltos en 1 mL de piridina anhidra y luego se adicion 1 mL de anh drido ac tico. La mezcla de reacci n se mantuvo en agitaci n constante a temperatura ambiente por 3 h, se detuvo la reacci n adicionando hielo y se trat con HCl 10 %, acetato de etilo y disoluci n saturada de NaHCO3de acuerdo al procedimiento convencional. La fase org nica fue secada (Na2SO4), concentrada y el derivado acetilado fue purificado mediante cromatograf a preparativa en capa delgada, usando como sistema de eluci n una mezcla de hexano:acetato de etilo 4:1.
Aceite amarillo (59 mg) Rf 0.42 (hexano:AcOEt, 7:3)
Resultados y Discusiones 55 RMN1H (CDCl3, 300 MHz); 6.85 (ddd,J= 0.8, 10.8, 17.2 Hz, H-14); 5.43 (t,J= 7.4 Hz, H-12);
5.29 (dd,J= 9.8, 10 Hz, H-6 ); 5.19 (d,J= 16.8 Hz, H-15); 5.10 (dd,J= 1.5, 10.8 Hz, H-15); 4.90 (d, J= 10.0 Hz, H-7 ); 2.45 (m, H-11a); 2.24 (m, H-11b); 2.06 (s, H3C(2 )-Ac); 2.01 (s, H3C(2 )- Ac); 1.78 (d,J= 1.2 Hz, H-16); 1.70-1.18 (m, H-1a, H-2a, H2b, H-9, H-5, H-3a, H-3b); 1.22 (s, H- 17); 0.97 (s, H-18); 0.95 (s, H-20); 0.95-0.97 (m, H-1b); 0.88 (s, H-19)
OH
OH OH H OH
OH O
H
15 16
9 5 7 3
1
20 17
19 18 11
13
OH
OR OH H O
118 117
118c R =
KOH/MeOH -BrC6H4COCl
10% Py
OH
OAc OAc H
OH
OMTPA OH
H
118d (R)-MTPA 118e (S)-MTPA
118a
9 5 7
17
A A / Py ( )-MTPA-Cl/ Py ( )-MTPA-Cl/ Py
O
Br
118b R =
O ( )-C6H4CHCHCOCl
6.2.6 7-p-Bromoben o laustroinulina (118b)
60 mg de 118 fueron sometidos a hidr lisis en presencia de KOH 10 % (20 mL) obteni ndose 36 mg de 117. Posteriormente, 117 fue sometido al proceso de esterificaci n con cloruro de p- bromobenzo lo en piridina anhidra por 24 h y tratado de manera convencional hasta la obtenci n del derivado p-bromobenzoilado118b(29 mg). Cabe se alar que, el producto mayoritario se esterific preferencialmente en la posici n 7, mientras que el alcohol de la posici n 6 no reacciona, presumiblemente por el impedimento est rico que origina el grupop-bromobenzoato de la posici n 7. El producto fue purificado mediante CCDP usando Hex:(CH3)2CO (7:3) como eluyente.
S lido blanco amorfo
UV [c1.98 X 10-5, EtOH] 204 (log 4.48), 215 (log 4.17), 241 (log 4.62) DC [c1.98 X 10-5, EtOH] [ ]247+ 19 726, [ ]231 5 464, [ ]208+ 5 914
RMN 1H (CDCl3, 300 MHz); 7.94 (2H, d, J = 9.0 Hz, AA MM ), 7.58 (2H, d, J = 8.7 Hz, AA MM ), 6.86 (ddd,J= 17.4, 10.8, 0.9 Hz, H-14), 5.45 (t,J= 7.8 Hz, H-12), 5.20 (d, J= 17.1 Hz, H-15trans), 5.10 (dt, J= 10.8, 1.8 Hz, H-15cis), 5.02 (d, J= 9.9 Hz, H-7), 3.89 (dd, J= 11.2, 9.8 Hz, H-6), 2.50 (dt,J= 15.6, 5.1 Hz, H-11a), 2.23 (dt,J= 16.2, 6.0 Hz, H-11b), 1.78 (d,J= 1.2 Hz, H3-16), 1.32 (s, H3-17), 1.27 (d,J= 11.1 Hz, H-5), 1.17 (s, H3-18), 1.01 (s, H3-20), 0.96 (s, H3-19) EM-FAB+: m/z (% intensidad relativa) 504 (2, M+), 505 (3), 507 (2), 487 (7, 505 - H2O), 489 (3, 507 H2O), 183 (100, [C7H4OBr]+), 185 (97, [C7H4OBr]+), 81 (67, [C6H9]+).
6.2.7 7-(E)-Cinamo laustroinulina (118c): 118 (40 mg) fue sometido a una hidr lisis alcalina para obtener 117 (31 mg), el cual fue posteriormente esterificado con cloruro de (E)- cinamo lo en piridina anhidra hasta obtener 118c (23 mg). UV [c 4.42 X 10-5, MeOH] 238 (log 4.34), 277 (log 4.37); DC [c 4.42 X 10-5, MeOH] [ ]276 + 6 737, [ ]266 + 6 961, [ ]237 15 667, [ ]217 10 710; RMN1H (CDCl3, 300 MHz) 7.74 (d, J = 15.9 Hz, AB), 7.53 (m, 2H), 7.39 (m, 2H), 6.87 (ddd, J= 17.1, 10.5, 0.6 Hz, H-14), 6.52 (d,J= 16.2 Hz, AB), 5.46 (t, J= 7.5 Hz, H-12), 5.19 (dd,J= 17.1, 0.6 Hz, H-15trans), 5.10 (dt,J= 10.8, 1.5 Hz, H-15cis), 4.90 (d,J= 9.9 Hz, H-7 ), 3.85 (dd,J= 11.1, 9.9 Hz, H-6 ), 2.50 (dt,J= 15.9, 6.0 Hz, H-11a), 2.23 (dt,J= 16.2, 5.7 Hz, H- 11b), 1.78 (d,J= 1.5 Hz, H3-16), 1.27 (s, H3-17), 1.17 (s, H3-18), 1.02 (s, H3-19), 0.96 (s, H3-20)
6.2.8 Preparaci n de los derivados esterificados (R)-MTPA y (S)-MTPA de 117 (118d y 118e): 35 mg de 118 fueron hidrolizados con KOH/MeOH al 10 % hasta obtenci n de 117 (16 mg). Luego, 2.0 mg de117fueron sometidos a reaccionarin situ(en un tubo de RMN) con 6 L del cloruro (S)-(+)- -metoxi- -(trifluorometil)fenilacetilo en 0.5 mL de piridina-d5 anhidra [41]. El tubo de RMN fue agitado cuidadosamente para mezclar la muestra y posteriormente, el curso de la reacci n fue monitoreada cada 2 h, mediante RMN 1H, hasta que el ster derivado (R)-MTPA (118d) de117 fue totalmente formado. RMN1H (Piridina-d5, 300 MHz) 7.2619 (ddd,J= 17.31, 10.77, 0.78 Hz, H-14); 5.7948 (t,J= 7.64 Hz, H-12); 5.7794 (d,J= 9.24 Hz, H-7 ); 5.2552 (dd,J= 17.46, 2.37 Hz, H-15trans); 5.1547 (dt, J= 10.89, 1.59 Hz, H-15cis); 4.0128 (dd, J= 10.0, 8.97 Hz, H-6 ); 2.9314 (m, H.11a); 2.4171 (m, H-11b). El procedimiento descrito para118d, fue aplicado para la formaci n del ster derivado (S)-MTPA (118e) de 117 a partir de la mezcla de
Resultados y discusiones 57 reacci n del cloruro (R)-( )- -metoxi- -(trifluorometil)fenilacetilo (6 L), piridina-d5 anhidra (0.5 mL) y117 (2.0 mg). RMN 1H (Piridina-d5, 300 MHz) 7.1784 (ddd,J= 17.11, 10.83 Hz, H-14);
5.7429 (t, J= 7.6 Hz, H-12); 5.8456 (d, J = 9.9 Hz, H-7 ); 5.2474 (dd, J= 17.13 Hz, H-15trans);
5.1285 (dt, J = 10.86 Hz, H-15cis); 4.2793 (dd, J = 11.0, 10.83 Hz, H-6 ); 2.8732 (m, H.11a);
2.3764 (m, H-11b). El espectro de RMN 1H proporcion evidencia s lo de se ales debidas a la esterificaci n preferencial del alcohol de la posici n 7. En los espectros de RMN1H de118dy118e se observaron se ales debidas al cloruro y al cido de Mosher.
6.2.9 Isopropilidendioxiaustroinulina (119)
A partir de la fracci n G se aisl el artefacto acet nido derivado del producto natural austroinulina (117), formado durante el proceso de separaci n y purificaci n por cromatografia en columna en la cual se us (CH3)2CO, CHCl3y Hex como sistema de eluci n. Cabe se alar que no se ha reportado previamente en la literatura este tipo de artefacto derivado de diterpenos de tipo labdano.
S lido blanco (28 mg)
Rf 0.45 (hexano:acetona, 7:3) Pf 106-108 C
IR CHCl3 max3460 (OH); 2985, 2921, 2874; 1230 (C-O); 1642, 1091, 896cm-1(C=C) [ ]D25+ 40 (c= 0.075, MeOH)
RMN1H (CD3COCD3, 500 MHz); 6.95 (ddd,J= 1.0, 11.0, 17.5 Hz, H-14); 5.48 (t,J= 7.0 Hz, H- 12); 5.13 (dd,J= 1.0, 17.0 Hz, H-15); 5.03 (dt, J= 10.8, 1.5 Hz, H-15); 3.61 (dd,J= 9.0, 11.0 Hz, H-6 ); 3.49 (OH); 3.42 (d, J= 9.5 Hz, H-7 ); 2.50 (m, H-11a); 2.27 (m, H-11b); 1.74 (d, J= 0.5 Hz, H-16); 1.64-1.58 (2H, m, H-1a, H-2a); 1.42-1.34 (3H, m, H-2b, H-9 , H-3a); 1.35 (d, J = 0.5 Hz, H3-2 ); 1.33 (d, J= 0.5 Hz, H3-3 ); 1.26 (d,J= 11.5 Hz, H-5 ); 1.16 (m, H-3b); 1.18 (s, H-17);
1.04 (s, H-18); 0.97 (s, H-20); 0.96 (s, H-19); 0.93 (d, H-1b)
RMN13C (CD3COCD3, 125 MHz); 135.68 (C-12), 135.04 (C-14), 130.48 (C-13), 113.48 (C-15), 108.89 (C-1 ), 90.76 (C-7), 76.62 (C-6), 74.99 (C-8), 61.51 (C-9), 56.69 (C-5), 42.69 (C-3), 41.46 (C-10), 40.84 (C-1), 35.33 (C-18), 34.12 (C-4), 27.58 (C-3 ), 27.06 (C-2 ), 22.98 (C-11), 22.49 (C- 19), 19.97 (C-16), 19.04 (C-2), 18.60 (C17), 16.98 (C-20)
EM IE: m/z (% Intensidad relativa), 362 (1, M+), 361 (1, [M H]+), 344 (10, [M H2O]+), 286 (10), 195 (65), 137 (50), 81 (82), 43 (100), 41 (42), 18 (12)
EM-AR-FAB+: Observado: 363.2889; Calculado: 363.2899, [M + H]+).
OH
O O
H
15 16
9 5 7 3
1
20 17
19 18 11
13
119
6.2.10 trans-Deshidrocrotonina (116) [10a, 10b]
S lido blanco amorfo (32 mg) Rf 0.32 (Hex:AcOEt, 75:25) Pf: 135 C
[ ]D25+ 13 (c= 0.075, CHCl3)
IR CHCl3 max2958, 2932, 2863; 1753 (C=O anillo -lactona); 1664 (C=O , -insaturado); 1504, 873 (anillo furano); 1154 (C-O); 1622 (C=C); 755cm-1
RMN 1H (CDCl3, 300 MHz); 7.45 (2H, m, H-15, H-16); 6.41 (dd, J= 1.0, 1.7 Hz, H-14); 5.89 (quint, J= 1.0 Hz, H-3); 5.43 (t,J= 8.6 Hz, H-12 ); 3.18 (t,J= 1.2, 11.5 Hz, H-5 ); 2.54 (dd,J= 3.0, 15.6 Hz, H-1 ); 2.43 (dd, J= 13.9, 8.6 Hz, H-11a); 2.36 (dd, J= 13.8, 8.7 Hz, H-11b); 2.26 (dquint,J= 12.9, 3.0 Hz, H-6a); 2.19 (dd,J= 14.1, 15.6 Hz, H-1 ); 1.97 (t,J= 1.0 Hz, H3-18); 1.85 (m, H-7a); 1.80 (ddd, J = 3.0, 10.4, 13.9 Hz, H-10 ); 1.66-1.69 (m, H-7b); 1.66-1.69 (m, H-8 );
1.17 (m, H-6b); 1.16 (d,J= 6.6 Hz, H3-17)
RMN13C (CDCl3, 75 MHz); , asignaci n: 197.82 (C-2), 177.00 (C-20), 166.14 (C-4), 144.30 (C- 15), 139.32 (C-16), 126.69 (C-3), 125.10 (C-13), 107.97 (C-14), 72.35 (C-12), 51.40 (C-9), 46.20 (C-10), 41.70 (C-8), 40.49 (C-11), 39.72 (C-1), 39.54 (C-5), 30.08 (C-7), 28.15 (C-6), 22.04 (C-18), 17.61 (C-17)
EM-IE+:m/z(int. rel. %) 314 (45,M+); 161 (100); 134 (20); 121 (45); 95 (17); 91 (12); 77 (10); 67 (7); 41 (10)
Resultados y discusiones 59
H
CH3
O
1
3 5 7
9 11 13 15
16
17 20
18 H O
O O H
116
6.2.11 Me cla de cajucarin lida (122) [10c] y 15-epi-cajucarin lida (123)
De las fracciones H-I se aisl y purific un s lido color beige que se revela como una mancha negra bien definida bajo la l mpara de UV254nmy que revela caf tenue con sulfato c rico de amonio.
S lido color beige (5.3 mg) Rf 0.25 (Hex:(CH3)2CO, 3:2) Pf 238-240 C
UV [c5.78 X 10-5M, MeOH] 205 (log 4.15), 235 (log 4.60)
DC [c5.78 X 10-5M, MeOH] [ ]324 8 399, [ ]271+ 2 506, [ ]243 41 300, [ ]222+ 56 754
IR CHCl3 max 3273 (OH); 2962, 2859; 1753 (C=O anillo -lactona); 1661 (C=O , -insaturado);
1213, 1107 (C-O); 1620 (C=C); 755cm-1
RMN1H (CDCl3:DMSO-d63:1, 300 MHz); 7.08 (2H, t,J= 1.2 Hz, H-14 , H-14); 6.24 (t,J= 1.2 Hz, H-15 ); 6.14 (dd,J= 1.2, 1.8 Hz, H-15); 5.87 (2H, sa, H-3 , H-3); 5.19 (2H, m, H-12 , H-12);
3.09 (2H, sa, H-5 , H-5); 2.60 (m, H-11a); 2.58 (m, H-1a); 2.51 (m,J= 13.8, 8.5 Hz, H-11a ); 2.34 (2H, m, H2-1 ); 2.29 (m, H-11b ); 2.23 (2H, m, H-6a , H-6a); 2.16 (m, H-11b); 2.12 (m, H-1b); 1.97 (6H, d,J= 1.0 Hz, H3-18 , H3-18); 1.82 (2H, m, H-7a , H-7a); 1.77 (2H, m, H-10 , H10); 1.70 (2H, m, H-8 , H-8); 1.68 (2H, m, H-7b , H-7b); 1.19 (2H, td,J= 12.6, 3.3 Hz, H-6b , H6b); 1.13 (3H, d, J= 6.6 Hz, H3-17); 1.12 (3H, d,J= 6.6 Hz, H3-17 )
RMN1H (CDCl3:DMSO-d63:1, 500 MHz); 7.09 (t,J= 1.0 Hz, H-14 ) 7.08 (t,J= 1.2 Hz, H-14);
6.24 (t,J= 1.2 Hz, H-15 ); 6.14 (dd,J= 2.0, 1.5 Hz, H-15); 5.87 (2H, dsept,J= 6.0, 1.5 Hz, H-3 , H-3); 5.18 (2H, ddd, J= 10.5, 9.0, 2.0 Hz, H-12 , H-12); 3.11 (2H, m, H-5 , H-5); 2.61(4H, m, H- 11a , H-11a, H2-1); 2.38 (dd, J = 15.5, 2.5 Hz, H-1a ); 2.30 (dd, J = 14.0, 9.0 Hz, H-11b ); 2.26 (2H, dquint, J= 13.5, 3.0 Hz, H-6a , H-6a); 2.17 (dd,J= 14.5, 9.0 Hz, H-11b); 2.09 (m, H-1b );
1.97 (6H, d, J = 1.5 Hz, H3-18 , H3-18); 1.84 (2H, dt, J= 13.0, 7.0 Hz, H-7a , H7a); 1.78 (2H, m, H-10 ,H-10); 1.72-1.65 (4H, m, H-8 , H-8, H-7b , H-7b); 1.19 (2H, m, H-6b , H-6b); 1.13 (3H, d,J
= 6.0 Hz, H3-17 ); 1.12 (3H, d, J= 6.5 Hz, H3-17)
RMN13C (CDCl3:DMSO-d6 3:1, 75 MHz); 197.53 (C-2 ), 197.29 (C-2), 176.46 (C-20 ), 176.40 (C-20), 168.50 (C-16 , C-16), 165.85 (C-4 ), 165.60 (C-4), 144.56 (C-16 ), 144.24 (C-14), 136.24 (C-13 ), 135.95 (C-13), 126.62 (C-3 ), 126.54 (C-3), 98.37 (C-15 ), 98.18 (C-15), 71.54 (C-12 , C- 12), 50.89 (C-9 ), 50.72 (C-9), 45.52 (C-10 , C-10), 41.54 (C-8 ), 41.33 (C-8), 39.59 (C-1 ), 39.41 (C-5 , C-5), 39.11 (C-1), 38.56 (C-11 ), 38.19 (C-11), 29.92 (C-7 ), 29.80 (C-7), 27.93 (C-6 ), 27.87 (C-6), 21.81 (C-18 , C-18), 17.34 (C-17 ), 17.24 (C-17)
RMN13C (CDCl3:DMSO-d63:1, 125 MHz); 197.15 (C-2 ), 196.87 (C-2), 176.04 (C-20 ), 175.98 (C-20), 168.50 (C-16 , C-16), 165.44 (C-4 ), 165.25 (C-4), 144.16 (C-16 ), 143.82 (C-14), 135.68 (C-13 ), 135.56 (C-13), 126.20 (C-3 ), 126.13 (C-3), 97.95 (C-15 ), 97.76 (C-15), 71.11 (C-12 , C- 12), 50.49 (C-9 ), 50.32 (C-9), 45.10 (C-10 , C-10), 41.11 (C-8 ), 40.92 (C-8), 39.16 (C-1 ), 39.00 (C-5 ), 38.94 (C-5), 38.69 (C-1), 38.14 (C-11 ), 37.77 (C-11), 29.47 (C-7 ), 29.36 (C-7), 27.47 (C- 6 ), 27.42 (C-6), 21.43 (C-18 ), 21.40 (C-18), 16.91 (C-17 ), 176.82 (C-17)
EM-FAB+:m/z(int. rel. %) 693 (4, [2(M) + H]+), 347 (82, [M+ H]+), 259 (51), 243 (100), 154 (80), 136 (68), 91 (32), 77 (30), 55 (25), 43 (18), 41 (21)
H
CH3 O
HO
1
3 5 7
9 11 13 15
16
17 20
18 H O
O
H CH3
O HO
1
3 5 7
9 11 13 15
16
17 20
18 H O
O
O O
O O
H H
+
122 123 6.2.12 Marrubiagenina (121) [36a]
S lido blanco (3.6 mg) Rf 0.37 (Hex:(CH3)2CO, 7:3) Pf 174-175 C
[ ]D25+ 54.4 (c= 0.18, CHCl3), ([ ]D25 60.5,c0.9, CHCl3, [44]), ([ ]D25 58, c2.3, CHCl3, [43])
Resultados y discusiones 61 UV (c 9.03 x 10-5 M, MeOH) 225 (log 4.38); [MeOH, 208 nm, log 4.30, [44]]; [CH3CN, 209 nm, log 4.30, 36a].
DC (MeOH,c9.03 x 10-5M) [ ]210 13 240; [ ]216 9 946; [ ]242+ 18 886; [(MeOH, c3 x 10-5M) [ ]205 + 5,300; [ ]215 0; [ ]235 14,900, [44]]; [(MeOH, : 243 (+ 6.38) = [ ]243+ 21,054; 215 (- 2.16) = [ ]215 7,128, [36a].
IR CHCl3 max 3500-2636, 1779 (CO-OH); 1746 (C=O anillo lactona); 1679 (C=O cido , - insaturado); 1635 (C=C); 1267, 1173 (C-O)cm-1
RMN1H (CDCl3, 500 MHz); 6.82 (t,J= 4.0 Hz, H-3); 5.86 (quint, J= 1.5 Hz, H-14); 4.76 (2H, d, J= 2.0 Hz, H-16); 2.75 (ddd,J= 17.0, 11.5, 3.5 Hz, H-6 ); 2.40 (ddd, J= 13.0, 9.0, 4.0 Hz, H- 2 ); 2.27 (3H, m, H-2 , H2-12); 2.06 (m, H-1 ); 1.79 (dd, J= 15.5, 9.0 Hz, H-1 ); 1.73 (dd, J = 10.5, 7.3 Hz, H-11a); 1.55 (dd, J= 10.5, 7.0 Hz, H-11b); 1.47 (m, H-8 ); 1.40 (dd,J= 6.5, 1.0 Hz, H-10 ); 1.32 (dddd,J= 16.0, 13.5, 3.0 Hz, H-7 ); 1.25 (3H, s, H3-19 ); 1.18 (dddd,J= 16.0, 14.0, 2.0 Hz, H-7 ); 1.11 (ddd,J= 16.5, 13.5, 3.0 Hz, H-6 ); 0.83 (3H, s, H3-20 ); 0.78 (3H, d,J= 6.5 Hz, H3-17 )
RMN13C (CDCl3, 125 MHz); , asignaci n: 173.98 (C-15), 171.74 (C-18), 170.88 (C-13), 142.05 (C-3), 137.25 (C-4), 115.13 (C-14), 73.05 (C-16), 45.48 (C-10), 40.26 (C-9), 37.38 (C-8), 36.74 (C- 6), 36.33 (C-5), 35.24 (C-11), 33.35 (C-19), 28.47 (C-7), 24.28 (C-2), 22.22 (C-12), 17.88 (C-20), 16.95 (C-1), 15.91 (C-17)
EM-IE+:m/z(int. rel. %) 332 (7,M+), 333 (2, [M+ H]+), 314 (67, [M H2O]+), 299 (100, [M CH3
H2O]+), 271 (15), 203 (31), 125 (22), 111 (19, C6H7O2+), 98 (16, C5H6O2+), 91 (19), 69 (24), 57 (23), 55 (23), 41 (17), 28 (14)
HRFABMS: Observado: 333.2076; Calculado: 333.2066, C20H29O4, [M + H]+).
H
CH3
CH3 CH3 O O
CO2H 1
3 5 7
9 11 13 15
16
17 20
19 18
121
6.2.13 5,7,4 -Trihidroxi-3-metoxiflavona o 3-O-metilkaempferol (120) [37]
S lido amarillo amorfo (61 mg) Rf 0.35 (Hex:(CH3)2CO, 3:2) Pf 268-270 C
IR CHCl3 max3494, 3125 (OH); 2942, 2732, 2588, 2492, 2378, 2101; 1650 (C=O , -insaturado);
1604; 1565, 1490, 1358, 1297, 1220, 1165, 1087, 1016, 837, 804, 717, 637, 576
RMN1H (CDCl3, 300 MHz); HO-C(5) no fue observado en la regi n de 11.0-12.0 ppm, 7.93 (dt, J= 8.79, 2.04 Hz, H-6 , H-2 ); 6.88 (dt,J= 9.09, 2.07 Hz, H-3 , H-5 ); 6.33 (d,J= 2.09 Hz, H-8);
6.14 (d, J= 1.77 Hz, H-6); 3.90 (3H, s, H3-CO-C(3), los cuales correlacionaron, en el experimento COLOC, con C-3 ( 139.42))
RMN13C (CDCl3, 75 MHz); 179.96 (C-4), 165.83 (C-7), 163.04 (C-5), 161.63 (C-4 ), 158.37 (C- 9), 157.97 (C-2), 139.42 (C-3), 131.39 (C-2 , C-6 ), 122.55 (C-1 ), 116.53 (C-3 , C-5 ), 105.87 (C- 10), 99.75 (C-6), 94.76 (C-8), 60.49 (CH3-O)
EM-FAB+:m/z(int. rel. %) 301 (100, [M+ H]+), 300 (35,M+), 286 (12, [M CH3]+), 154 (41), 136 (34), 121 (11), 107 (12), 89 (13), 77 (17), 55 (14), 41 (10)
O
O O HO
OH
OH
3
4'
7
5
1 1'
120
6.2.14 5,7,3 ,4 -Tetrahidroxi-3-metoxiflavona (124) [38 ] S lido blanco amorfo (5 mg)
Rf 0.21 (Hex:AcOEt, 1:1) Pf 254-256 C
IR CHCl3 max3399 (OH); 2918, 2850; 2256, 2128 (Ar); 1653 (C=O , -insaturado); 1600, 1362, 1302, 1174, 1025, 1002, 823, 762cm-1
RMN1H (CDCl3, 300 MHz); 12.58 (s, HO-C(5)); 7.69 (d,J= 2.2 Hz, H-2 ); 7.56 (dd,J= 8.4, 2.2 Hz, H-6 ); 6.97 (d,J= 8.4 Hz, H-5 ); 6.39 (d,J= 2.0 Hz, H-8); 6.29 (d,J= 2.2 Hz, H-6); 3.83 (3H, s, H3-CO-C(3))
Resultados y discusiones 63 RMN13C (CDCl3, 75 MHz); 177.98 (C-4), 163.38 (C-7), 161.46 (C-5), 156.26 (C-9), 155.17 (C- 2), 146.96 (C-4 ), 143.85 (C-3 ), 137.90 (C-3), 121.07 (C-1 ), 120.73 (C-6 ), 114.88 (C-2 ), 114.61 (C-5 ), 104.50 (C-10), 98.43 (C-6), 93.16 (C-8), 59.20 (CH3-O)
EM-IE+:m/z(int. rel. %) 316 (21, M+), 315 (20, [M H]+), 111 (36), 97 (58), 83 (62), 71 (64), 69 (73), 57 (100), 55 (67), 43 (62), 28 (72), 18 (63)
124
6.2.15 -D-glucopiran sido de -sitosterilo (125) [39]
S lido blanco-verdoso (1.45 g)
RMN1H (Py-d5, 300 MHz); , (multiplicidad, J(Hz), asignaci n): 5.35 (t,J= 2.5 Hz, H-6), 5.04 (d,J= 7.69 Hz, H-1 ), 4.55 (dd,J= 2.5, 11.77 Hz, H-6 a), 4.40 (dd, J= 5.2, 11.77 Hz, H-6 b), 4.27 (2H, t, J= 7.50 Hz, H-3 , H-4 ), 4.04 (dd,J= 7.69, 7.50 Hz, H-2 ), 3.98 (m, H-3), 3.96 (m, H-5 ), 2.72 (ddd,J= 1.98, 4.69, 12.94 Hz, H-4a), 2.49 (ddd,J= 1.98, 12.94, 12.94 Hz, H-4b), 2.14 (m, H- 2a), 1.98 (dd, J= 4.3, 12.37 Hz, H-12), 1.95 (ddd, J= 16.0, 2.5, 7.0 Hz, H-7b), 1.85 (m, H-16b), 1.75 (m, 2b), 1.72 (m, H-1a), 1.68 (m, H-25), 1.60 (ddd, J= 2.5, 7.0, 16.0 Hz, H-7a), 1.57 (m, H- 15b), 1.42 (2H, m, H-11), 1.40 (2H, m, H-20, H-22a), 1.36 (m, H-8), 1.30 (2H, m, H28), 1.25 (3H, m, H-16a, H-23), 1.10 (3H, m, H-12b, H-17, H-22b), 1.05 (m, H-15a), 1.00 (m, H-24), 0.98 (m, H- 1b), 0.98 (d,J= 6.5 Hz, H-21), 0.95 (m, H-14), 0.94 (s, H-19), 0.88 (d,J= 7.5 Hz, H-29), 0.86 (d,J
= 7.0 Hz, H-27), 0.85 (m, H-9), 0.84 (d,J= 7.0 Hz, H-26), 0.66 (s, H-18)
RMN13C (Py-d5, 75 MHz); , asignaci n: 140.95 (C-5), 121.90 (C-6), 102.57 (C-1 ), 36.36 (C- 20), 12.13 (C-29), 78.54 (C-3 ), 78.13 (C-5 ), 78.38 (C-3), 75.27 (C-2 ), 71.67 (C-4 ), 62.82 (C-6 ), 50.34 (C-9), 56.25 (C-17), 56.83 (C-14), 46.04 (C-24), 32.04 (C-8), 34.20 (C-22), 39.31 (C-4), 37.46 (C-1), 42.47 (C-13), 36.08 (C-10), 28.50 (C-16), 32.15 (C-7), 18.99 (C-21), 30.21 (C-2), 26.40 (C-23), 24.48 (C-15), 39.95 (C-12), 21.26 (C-11), 19.29 (C-30), 23.28 (C-28), 29.47 (C-25), 11.95 (C-18), 19.39 (C-19), 19.20 (C-26), 19.94 (C-27)
O
22 23
3 6 1
16 14 10
19
18
11
26 27 28
O OH
HO HO
OH
2' 1' 4' 6'
125
6.2.16 Sacarosa (126) [40]
S lido (400 mg) Pf 179-182 C
IR (KBr), max: 3382, 2921, 1647, 1417, 1145, 1049, 925cm-1
RMN1H (300 MHz, CD3OD), : 5.38 (d,J= 3.84 Hz, H-1), 4.07 (d,J= 8.3 Hz, H-3 ), 4.01 (t,J= 8.3 Hz, H-4 ), 3.82 (m, H-5), 3.80 (2H, m, H-6), 3.75 (3H, m, H-1 , H-5 ), 3.70 (m, H-3), 3.60 (2H, m, H-6), 3.42 (dd,J= 3.5, 9.5 Hz, H-2), 3.34 (m, H-4)
RMN13C (75 MHz, CD3OD), : 105.32 (C-2 ), 93.65 (C-1), 83.75 (C-5 ), 79.36 (C-3 ), 75.71 (C- 4 ), 74.61 (C-3), 74.38 (C-5), 73.19 (C-2), 71.34 (C-4), 64.03 (C-6 ), 63.39 (C-1 ), 62.219 (C-6) EM-FAB+ m/z(int. rel.): 365 [M + Na]+ (6), 329 (30), 307 (34), 289 (13), 176 (42), 154 (100), 136 (61), 107 (12), 89 (11), 77 (10), 65 (4), 23 (4)
O
O H OH
HO HO
OH
O
OH OH
HO
OH
1 2 6
2' 1' 5'
4' 4
126
Resultados y discusiones 65