Beruflich Dokumente
Kultur Dokumente
Tetrahedron Letters
journal homepage: www.elsevier.com/locate/tetlet
National Center for Natural Products Research, School of Pharmacy, The University of Mississippi, University, MS 38677, USA
Department of Pharmacognosy, School of Pharmacy, The University of Mississippi, University, MS 38677, USA
c
Ege University, Faculty of Science, Biology Department, Izmir 35100, Turkey
b
a r t i c l e
i n f o
Article history:
Received 19 September 2013
Revised 4 November 2013
Accepted 10 November 2013
Available online 16 November 2013
a b s t r a c t
A new indole alkaloid, 7b-hydroxy-7H-mitraciliatine (1) and a new oxindole alkaloid, isospeciofoleine (2)
together with nine known alkaloids were isolated from Mitragyna speciosa and characterized by NMR, CD,
and MS spectroscopic data analyses. The 1H and 13C NMR spectroscopic data of isospeciofoline (3), isorotundifoline (4), paynantheine (5), and 3-isopaynantheine (6) were also reported for the rst time.
2013 Elsevier Ltd. All rights reserved.
Keywords:
Mitragyna speciosa
Rubiaceae
Indole/oxindole
7b-Hydroxy-7H-mitraciliatine
Isospeciofoleine
Compound 5 (108 mg) was obtained as an upper band from preparative thin layer chromatography (PTLC) [silica gel plate
(0.5 mm), hexanes/acetone/NH4OH, (208:90:1)] of fraction 5EA.
The contents of the lower band were applied to PTLC [silica gel
plate (0.5 mm), CHCl3/EtOAc (7:3)], followed by reverse phased
HPLC [C-18 silica, MeOH/H2O, 65:35] to purify compounds 2
OMe
9
10
11
12
13
9
5
N4
15
14
MeO
22
OH
OH
OH 6
11
21
20
16
19
10
12
13
18
N
H
OMe
14
MeO
17
OH
N
H
21
20
15
19
OMe
MeO
OMe
O
Isospeciofoline (3),
OMe
N
H
MeO
N
H
OMe
Isorotundifoline (4)
Paynantheine (5)
OMe
O
Isospeciofoleine (2)
MeO
17
OMe
N
H
18
16
22
7-Hydroxy-7H-mitraciliatine (1)
MeO
OMe
O
3-Isopaynantheine (6)
370
Table 1
H NMR spectroscopic dataa for compounds 16 (500 MHz, d in ppm, CDCl3)
No.
1
dH, mult. (J in Hz)
2
dH, mult. (J in Hz)
3
dH, mult. (J in Hz)
4
dH, mult. (J in Hz)
5
dH, mult. (J in Hz)
6
dH, mult. (J in Hz)
3
5
4.24,
3.51,
2.70,
2.49,
2.01,
6.70,
7.27,
7.21,
2.37,
2.09
3.03,
7.34,
0.71,
d (5.5)
br t (14.1)
br d (14.1)
br d (14.1)
t d (14.1, 4.1)
br d (8.0)
t (8.0)
br d (8.0)
t d (13.1, 5.9)
2.61,
3.42,
2.58,
2.43,
2.21
6.60,
7.08,
6.43,
2.14
1.26
2.92,
7.32,
0.90,
3.01,
3.35,
2.61,
2.35,
2.06,
6.46,
6.97,
6.31,
1.43,
1.35,
3.21,
7.32,
0.77,
m
m
3.23,
3.07
2.57,
3.17,
2.99
6.45,
6.99,
6.87,
2.09,
1.96,
2.75,
7.33,
5.00,
4.96,
5.58,
4.44, br s
3.22
1.30,
0.87,
2.03
2.44,
2.06
3.83,
3.81,
3.69,
2.69,
3.42,
2.61
2.43,
2.18
6.57,
7.06,
6.41,
1.69
1.38
2.61
7.20,
4.98,
4.95,
5.50,
br d (7.9)
t (7.9)
br d (7.9)
q (12.1)
d t (12.6, 3.3)
t d (11.8, 3.8)
s
dd (17.2, 2.0)
dd (10.3, 2.0)
ddd (17.9, 10.3, 8.0)
3.04
3.02,
2.27,
3.86,
3.75,
3.70,
8.18,
br d (10.1)
br t (11.2)
s
s
s
s
6
10
11
12
14
15
17
18
19
20
21
9-OMe
17-OMe
22-OMe
NH
OH
a
br t (11.9)
s
t (7.2)
br d (10.6)
s
s
s
br d (11.4)
br t (9.8)
br t (12.1)
br d (8.5)
t (8.5)
br d (8.5)
s
br d (18.0)
br d (10.8)
dt (18.0, 10.8)
br d (11.3)
t d (9.0, 2.5)
br t (9.0)
br t (10.3)
br d (8.0)
t (8.0)
br d (8.0)
d t (13.1, 2.8)
s
t (7.0)
dd (12.0, 2.9)
t d (9.1, 3.0)
m
ddd (13.7, 10.7, 3.2)
d t (13.2, 8.5)
br d (7.9)
t (7.9)
br d (7.9)
t d (13.1, 12.3, 6.7)
br d (13.5)
t (6.3)
s
t (7.4)
1.14
2.90, m
3.21, br d (10.2)
2.13, t (11.0)
1.66
1.26
1.64
3.32, br d (11.9)
2.36, br d (11.9)
3.70, s
3.59, s
9.01, s
11.86, s
3.64, s
3.60, s
8.67, s
11.74, s
3.72, s
3.59, s
9.09, s
br d (10.7)
d t (11.0, 5.6)
m
3.18
2.84,
6.47,
7.02,
6.95,
2.45,
1.99,
2.36,
7.26,
4.91,
4.83,
5.34,
br d (14.5)
br d (7.9)
t (7.9)
br d (7.9)
t d (13.6, 5.7)
br d (13.6)
br t (12.5)
s
br d (17.8)
br d (9.8)
dt (17.8, 9.8)
2.95, p (8.2)
2.63, br d (7.5)
3.88,
3.72,
3.65,
8.19,
s
s
s
s
Overlapped.
(2 mg), 3 (11 mg), and 4 (77 mg). Speciogynine (45 mg) was puried by PTLC [silica gel plate (0.5 mm), hexanes/acetone/NH4OH,
(208:90:1)] of fraction 5I. Speciociliatine (650 mg) and corynoxine
(44 mg) were obtained from fractions 7 and 8 by CC [silica gel
(3600 100 ), CHCl3/MeOH (49:1)] followed by PTLC [silica gel plate
(0.5 mm), hexanes/acetone/NH4OH, (180:120:1)]. Compound 6
(28 mg) was obtained from fraction 8 by CC [silica gel (40
0.75), hexanes/acetone/NH4OH, (180:120:1)] and compound 1
(17 mg) was puried from fractions 10 and 11 (138 mg) by PTLC
[silica gel plate (0.5 mm), CHCl3/MeOH/NH4OH, (28:15:1)].
Compound 16 was obtained as a yellow solid (a22
D 149.0 (c
0.13, MeOH)). The molecular formula of compound 1 was inferred
to be C23H30N2O5 from a protonated ion [M+H]+ at m/z 415.2224 in
the HR-ESI-ToF-MS (calcd for C23H31N2O5, 415.2233). The UV spectrum showed absorptions at 291, 240, and 220 nm. The IR spectrum exhibited absorption due to conjugated ester carbonyl
function at 1700 cm1. The 13C NMR spectrum exhibited 23 resonances which were differentiated as four methyl, ve methylene,
seven methine, and seven quaternary carbons. The characteristic
resonances for three methoxy groups [dH/dC 3.83 (s)/55.5
(9-OCH3), 3.81 (s)/61.6 (17-OCH3), and 3.69 (s)/51.4 (22-OCH3)],
a primary methyl [dH/dC 0.71 (t, J = 7.2 Hz)/11.2 (CH3-18)], an oxygenated olenic bond in conjugation with the oxo group [dH/dC
7.34 (s)/160.1 (CH-17) and dC 112.1 (C-16)], and an iminic quaternary carbon [dC 182.9 (C-2)] were found in the 1H and/or 13C NMR
spectra. The resonances for a 1,2,3-trisubstituted phenyl ring with
an ABC spin system [dH/dC 6.70 (br d, J = 8.0 Hz)/108.8 (CH-10),
7.27 (t, J = 8.0 Hz)/130.6 (CH-11), 7.21 (br d, J = 8.0 Hz)/114.3
(CH-12), dC 126.5 (C-8), 155.9 (C-9), and 154.6 (C-13)] and an oxygenated quaternary carbon [dC 82.1 (C-7)] along with ve aliphatic
methylenes and three aliphatic methines were also observed. The
1
H and 13C NMR data (See Tables 1 and 2) were assigned with
the help of HSQC, 1H1H COSY, and HMBC (Fig. 2) spectra. NMR
data analyses showed that 1 had the same planer structure as that
of 7-hydroxyspeciocilliatine.7 Literature reports indicated that the
13
C NMR resonances of an ethyl group were the key to determining
the conguration at C-20. The reported 13C NMR resonances of the
ethyl group are dC 18.920.5 (C-19) and dC 12.613.3 (C-18) for
C-20 S and dC 24.224.4 (C-19) and dC 11.211.3 (C-18) for C-20
R.7,8 Based on the chemical shifts of C-20 ethyl group [dC 24.2
(C-19) and dC 11.2 (C-18)], the conguration at C-20 in 1 was assigned to be R. The CD spectrum of 1 (Fig. 3) showed negative cotton effects at 316, 272 and 216 nm and found to be partially
comparable to that of 7-hydroxyspeciociliatine, for which negative
cotton effects at 307 and 256 nm and positive cotton effect at
230 nm were reported.7 The reverse cotton effects at lower absorbance could be due to the opposite congurations at C-20 in 1 and
7-hydroxyspeciociliatine.7 Consequently, the structure of 7b-hydroxy-7H-mitraciliatine (1) was established as shown in Figure 1.
Compound 29 was obtained as a yellow solid (a27
D 38 (c 0.05,
MeOH)). The molecular formula of compound 2 was inferred to
be C22H26N2O5 from a protonated ion [M+H]+ at m/z 399.1913 in
the HR-ESI-ToF-MS (calcd for C22H27N2O5, 399.1920). The UV spectrum showed absorptions at 290, 239, and 220 nm. The absorptions in the IR spectrum at 1623 and 1706 cm1 supported the
amide carbonyl and conjugated ester carbonyl functions. The 13C
NMR spectrum displayed 22 resonances, which were differentiated
as two methyl, ve methylene, eight methine, and seven quaternary carbons. The 1H and/or 13C NMR spectra of 2 showed characteristic resonances for two methoxy groups [dH/dC 3.70 (s)/61.4
(17-OCH3) and 3.59 (s)/51.0 (22-OCH3)], an oxygenated olenic
bond adjacent to an oxo group [dH/dC 7.20 (s)/159.7 (CH-17) and
dC 111.3 (C-16)], and an external double bond [dH/dC 5.50 (dt,
J = 18.0, 10.8 Hz)/138.5 (CH-19), 4.98 (br d, J = 18.0 Hz) and 4.95
(br d, J = 10.8 Hz)/116.2 (CH2-18)], a 1,2,3-trisubstituted phenyl
ring [dH/dC 6.57 (br d, J = 8.5 Hz)/111.6 (CH-10), 7.06 (t,
J = 8.5 Hz)/129.4 (CH-11), 6.41 (br d, J = 8.5 Hz)/101.0 (CH-12), dC
116.6 (C-8), 154.4 (C-9), and 140.6 (C-13)], a conjugated ester carbonyl carbon [dC 168.2 (C-22)], and an amide carbonyl carbon [dC
179.2 (C-2)]. The 1H and 13C NMR data assignment (See Tables 1
and 2) was done by analyses of the HSQC, 1H1H COSY, and HMBC
(Fig. 2) spectra. When the NMR spectroscopic data of 2 were compared with those of isospeciofoline (3), the resonances for an ethyl
group were found missing in 2 showing instead for an external
double bond. The sharp signals at dH 11.86 and dH 11.74 in the
1
H NMR spectra of 2 and 3 indicated a non-acidic phenol because
of intramolecular hydrogen bonding between C-9 hydroxyl and
lone pair of N-4 (Fig. 4), which ultimately supported the S
371
13
No.
1
dC, mult.
2
dC, mult.
3
dC, mult.
4
dC, mult.
5
dC, mult.
6
dC, mult.
2
3
5
6
7
8
9
10
11
12
13
14
15
16
17
18
19
20
21
22
9-OMe
17-OMe
22-OMe
182.9, C
54.4, CH
48.8, CH2
35.3, CH2
82.1, C
126.5, C
155.9, C
108.8, CH
130.6, CH
114.3, CH
154.6, C
28.2, CH2
33.7, CH
112.1, C
160.1, CH
11.2, CH3
24.2, CH2
39.5, CH
51.2, CH2
169.0, C
55.5, CH3
61.6, CH3
51.4, CH3
179.2, C
68.5, CH
53.0, CH2
34.0, CH2
56.9, C
116.6, C
154.4, C
111.6, CH
129.4, CH
101.0, CH
140.6, C
28.9, CH2
37.1, CH
111.3, C
159.7, CH
116.2, CH2
138.5, CH
41.8, CH
57.3, CH2
168.2, C
180.1, C
70.5, CH
53.6, CH2
33.6, CH2
57.0, C
117.0, C
154.3, C
111.5, CH
129.2, CH
101.4, CH
141.0, C
25.2, CH2
38.3, CH
110.7, C
160.6, CH
12.7, CH3
18.8, CH2
39.6, CH
54.5, CH2
168.8, C
179.9, C
63.6, CH
53.4, CH2
33.7, CH2
57.2, C
116.6, C
154.3, C
111.4, CH
129.3, CH
101.1, CH
141.1, C
31.4, CH2
30.9, CH
110.8, C
159.6, CH
12.1, CH3
24.0, CH2
38.8, CH
52.9, CH2
169.4, C
61.4, CH3
51.0, CH3
61.3, CH3
51.2, CH3
61.1, CH3
51.5, CH3
133.1, C
60.0, CH
53.0, CH2
23.8, CH2
107.5, C
117.4, C
154.4, C
99.6, CH
121.7, CH
104.4, CH
137.5, C
33.3, CH2
38.7a, CH
111.7a, C
159.9, CH
115.4, CH2
139.4, CH
42.7, CH
61.3, CH2
169.0a, C
55.3, CH3
61.5, CH3
51.3, CH3
130.8, C
53.9, CH
51.4, CH2
19.0, CH2
107.6, C
117.9, C
154.3, C
99.5, CH
121.8, CH
104.6, CH
137.2, C
31.1, CH2
33.9a, CH
111.6, C
159.7, CH
115.2, CH2
139.5, CH
43.0, CH
51.0, CH2
168.8, C
55.2, CH3
61.4, CH3
51.2, CH3
Broad signal.
OMe
OH
OH
N
N
MeO
N O
H
OMe
MeO
OMe
) correlations of 1 and 2.
Figure 4. Intramolecular hydrogen bonding in compounds 2 and 3.
372
7.
8.
9.
10.
11.
12.
13.
14.
NMR: see Table 2; HR-ESI-ToF-MS m/z: 415.2224 [M+H]+, (calcd 415.2233 for
C23H31N2O5).
Kitajima, M.; Misawa, K.; Kogure, N.; Said, I. M.; Horie, S.; Hatori, Y.; Murayama,
T.; Takayama, H. J. Nat. Med. 2006, 60, 2835.
Sakakibara, I.; Takahashi, H.; Terabayashi, S.; Yuzurihara, M.; Kubo, M.; Ishige,
A.; Higuchi, M.; Komatsu, Y.; Okada, M.; Maruno, M.; Biqiang, C.; Jiang, H. X.
Phytomedicine 1998, 5, 8386.
Isospeciofoleine (2). Yellow powder; a27
D 38.0 (c 0.05, MeOH); UV (MeOH) kmax
nm (log e): 290 (3.18), 239 (3.86), 220 (4.08); IR cm1: 3300, 1706, 1623, 1481,
1432, 1279, 1236, 1205, 1145, 1138, 1107; CD (MeOH) kmax nm (De): 214
(8.13), 243 (1.41), 216 (1.21), 293 (0.93); 1H NMR: see Table 1; 13C NMR: see
Table 2; HR-ESI-ToF-MS m/z: 399.1913 [M+H]+, (calcd 399.1920 for
C22H27N2O5).
Hemingway, S. R.; Houghton, P. J.; Phillipson, J. D.; Shellard, E. J. Phytochemistry
1975, 14, 557563.
Shellard, E. J.; Houghton, P. J.; Resha, M. Planta Med. 1978, 34, 253263.
Shellard, E. J.; Houghton, P. J.; Resha, M. Planta Med. 1978, 34, 2636.
Shellard, E. J.; Lala, P. K. Planta Med. 1978, 33, 6369.
Beckett, A. H.; Shellard, E. J.; Phillipson, J. D.; Lee, C. M. Planta Med. 1966, 14,
277288.