Alkaloids from Crinum moorei

Article abstract of DOI:10.1016/S0031-9422(00)00433-7

Thirteen alkaloids were isolated from Crinum moorei two of which are new. These are 3-[4′-(8′-aminoethyl)phenoxy] bulbispermine and mooreine. The structures of the new alkaloids were determined by spectroscopic methods.

Full text of DOI:10.1016/S0031-9422(00)00433-7

Phytochemistry 56 (2001) 637±640
www.elsevier.com/locate/phytochem
Alkaloids from Crinum moorei
a
b
a,
Esameldin E. Elgorashi , Siegfried E. Drewes , Johannes van Staden *
aResearch Centre for Plant Growth and Development, School of Botany and Zoology, University of Natal Pietermaritzburg,
Private Bag X01, Scottsville 3209, South Africa
bSchool of Chemical and Physical Sciences, University of Natal Pietermaritzburg, Private Bag X01, Scottsville 3209, South Africa
Received 23 February 2000; received in revised form 13 September 2000
Abstract
0
0
Thirteen alkaloids were isolated from Crinum moorei two of which are new. These are 3-[4 -(8 -aminoethyl)phenoxy] bulbis-
permine and mooreine. The structures of the new alkaloids were determined by spectroscopic methods. # 2001 Elsevier Science
Ltd. All rights reserved.
0
0
Keywords: Crinum moorei; Amaryllidaceae; Alkaloids; 3-[4 -(8 -Amino ethyl)phenoxy] bulbispermine; Mooreine
1
. Introduction
and COSY analysis, the resonance of H-3 and H-11
could be pinpointed at ꢀ 4.40 and 3.95, respectively. The
®rst is clearly coupled to H-1 and H-2 and also to the
methylene group at C-4 while the second is coupled to
the methylene group at 3.32. These were the only likely
positions to which the tyramine residue- whose presence
was clearly indicated by the A X system present at ꢀ
Crinum moorei (Amaryllidaceae) is one of 21 Crinum
species found in southern Africa (Verdoorn, 1973). Cri-
num species are used by Zulu healers against swelling of
the body and for urinary tract problems (Hutchings et al.,
1
996). We report here on the isolation of the new com-
2
2
0
0
pounds 3-[4 -(8 -aminoethyl)phenoxy] bulbispermine 1,
and 2 which we have designated as mooreine, systematic
name [1,3]dioxolo [4,5-j]pyrrolo [3,2,1-de] phenanthri-
dinum, 1,2,4,5,12b, 12c-hexahydro-2-hydroxy-12c-meth-
oxy (Tables 1 and 2). Also isolated from this species
were the known compounds crinine, undulatine (Vila-
domat et al., 1995a), 3-O-acetylcrinine (Campbell et al.,
1
namidine, epibuphanisine, epivittatine (Viladomat et al.,
1
1
1
6.73 and ꢀ 7.05 Ð could be attached. HMBC and
NOESY were not helpful and no connectivities were
0
0
observed between H-11 or H-3 and either C-6 and C-2
0
0
or C-7 and C-8 . Also no NOE e€ects were observed
between protons on the basic ring and the phenethyl
moiety. Acetylation of the compound resulted in a
down®eld shift of both H-3 and H-11 to around 5.48
and 4.97 ppm, respectively. This indicates that acetyla-
tion occurred at both positions which further proved by
the molecular ion of the acetate of 371 mass unit with
the loss of 60 and 43 mass units, respectively. The pos-
sibility of the compound being a mixture of two com-
pounds was rejected by developing bulbispermine and
tyramine on a tlc using the same solvent system used for
998), powelline, cherylline (Kobayashi et al., 1984), cri-
995b), 1-epideacetyl-bowdensine (Viladomat et al.,
996), lycorine, and 1-O-acetyllycorine (Evidente et al.,
983).
2
. Results and discussion
the puri®cation of 1. Tyramine has R value of 0.67
f
while bulbispermine R value of 0.59. From the above
f
1
The H NMR of 1 showed the typical singlets asso-
ciated with H-7, H-10, and the methylenedioxy group at
.86, 6.53 and 5.87, respectively. From the HETCOR
acetylation results, it is clear that the compound under-
gone degradation at some stage during storage or ace-
tylation. NOE studies did not reveal much useful
information but did show a good correlation between
H-11 exo and H-12 exo. Construction of an accurate
model using the NOE information showed, however,
that the 3- and 11-positions were not sterically crowded
6
*
Corresponding author. Tel.: +27-33-260-5130; fax: +27-33-260-
897.
E-mail address: rcpgd@botany.unp.ac.za (J. van Staden).
5
0
031-9422/01/$ - see front matter # 2001 Elsevier Science Ltd. All rights reserved.
PII: S0031-9422(00)00433-7

638
E.E. Elgorashi et al. / Phytochemistry 56 (2001) 637±640
and equally exposed to attack by a tyramine moiety. In
these circumstances, the tyramine residue was attached
at the more reactive allylic C-3. In addition, computer
simulation programme H NMR data, using Advanced
Chemistry Development programme (ACD) from Aldrich
Table 2
13C NMR data (CD
3
OD) for compound 1, and bulbispermine
C no.
1
Bulbispermine
1
0
C-1
C-9
157.4 s
148.4 s
147.9 s
137.7 s
137.4 d
131.0 d
130.9 s
127.1 s
125.2 d
116.7 d
108.1 d
104.5 d
102.4 t
81.3 d
68.6 d
67.7 d
63.9 t
148.3 s
147.9 s
137.7 s
137.4 d
1
998, supported the attachment at C-3, as indicated in
C-8
C-10a
C-1
Table 3. It is suggested that the linkage to C-3 is via the
phenolic group of the tyramine moiety. This is based on
the observation that in both plicamine and secoplica-
0
0
0
C-2 , C-6
C-4
À
0
mine (both bonded through N), C-8 resonates in the
C-6a
C-2
127.1 s
125.2 d
1
3
È
C spectrum at 49.0 and 50.0 ppm, respectively (Unver
et al., 1999) whereas in 1 the relevant chemical shift is at
4.0 ppm. In addition, the shift of the ethylene group in
0
C-3 , C-5
4
C-10
C-7
108.1 d
104.5 d
102.5 t
81.3 d
68.6 d
67.6 d
64.0 t
0
0
authentic tyramine (36.7 and 43.4 for C-7 and C-8 ,
respectively) correspond closely to that observed for the
corresponding group in compound 1. Furthermore, ace-
OCH
C-11
C-3
2
O
tylation of tyramine at both C-1 and NH showed only
2
C-4a
C-12
C-6
61.8 t
51.7 s
61.7 t
51.6 s
C-10b
0
C-8
44.0 t
37.9 t
34.7 t
0
C-7
C-4
34.7 t
Table 3
1
Computer simulation programme H NMR chemical shifts of Protons
at positions 3 and 11 for bulbispermine and bulbispermine with the
tyramine attached either to C-3 or C-11
Proton no.
Bulbispermine
Bulbispermine with tyramine
moiety attached to
C-3
C-11
3
1
-H
1-H
4.37
3.91
4.68
3.91
4.21
5.12
Table 4
1H NMR and 13C NMR (CD
Table 1
1H NMR data (CD
3
OD) for compound 2
3
OD) for compound 1, and bulbispermine
Atom no.
13C
1H
1
Bulbispermine
9
158.5 s
152.8 s
±
±
0
0
0
H-2 , H-6
7.05 d (J=8.5)
6.86 s
10
7
H-7
6.83 s
142.1 d
139.3 s
134.2 s
128.4
9.36 s
±
0
H-3 , H-5
6.73 d (J=8.6)
6.53 s
11a
7a
3a
3
H-10
H-2
H-1
6.48 s
±
6.24 dd (J=2.3, 10.3)
6.05 dd (J=1.2, 10.3)
5.87 s
6.23 dd (J=2.2, 10.35)
6.02 d (J=10.35)
5.86 s
±
127.6 d
106.1 d
105.6 t
100.8 d
79.4 s
6.45 m
7.63 s
6.38
7.77 s
±
OCH
H-3
2
O
8
OCH
11
4.40 m
4.3d dd (J=2,4,8)
4.22 d (J=16.9)
3.93 dd (J=2.9, 7)
3.68 d (J=16.9)
3.41 dd (1H, J=7, 13.69)
3.32 m (1H)
2
O
H-6a
H-11
H-6a
H-12
4.27 d (J=16.9)
3.95 dd (J=3.6,7)
3.72 d (J=16.9)
11c
2
72.0 d
68.9 d
57.1 t
4.61 m
5.31 m
5.00 m (2H)
11b
5
0
H-12
H-4a
3.32 m (2H)
3.32 m
3.20 m
1
54.8 t
5.49 ddd (0.69, 3.8, 5.3)
5.48 ddd (0.46, 1.14, 5.3)
3.32 m (2H)
0
0
2
2
2
H-8
H-7
H-4
2.90 t (J=7)
2.70 t (J=7)
2.02 m
4
OMe
26.5 t
48.0
2.03 m
3.32 s (3H)

E.E. Elgorashi et al. / Phytochemistry 56 (2001) 637±640
639
0
0
0
pared to 0.25 ppm down®eld shift of protons at C-3 and
.15 ppm dow®eld shift of protons at C-2 and C-6 com-
0
detailed analysis of the compound and the proposed
structure should be regarded as tentative.
0
0
C-5 . Protons attached to C-8 also shifted down®eld by
0 0
0
.28 ppm. Chemical shifts of C-2 , C-6 remained unaf-
0
0
fected while that of 3 , 5 were shifted down®eld by 6 ppm.
The compound gave no molecular ion in its HRMS.
However, it showed a fragment ion at 286.1058
3. Experimental
NMR spectra were recorded in CDCl and CD OD
3
3
(
permine after loss of the tyramine moiety.
C H NO ) which is the molecular ion of bulbis-
using TMS as internal standard at 500 and 200 MHz for
1H and 50 and 125 MHz for 13C. Chemical shifts are
recorded in ꢀ units and coupling constants (J) in Hz.
Mass spectra were recorded on a Kratos MS 80 RF
double-focussing magnetic sector instrument at 70 eV.
Silica gel Merck (230±400 mesh) was used for VLC .
Silica gel 60 F254 analytical and prep. TLC (2 mm) were
used for additional separation. Spot detection by UV
light (254) and Dragendor€'s reagent.
1
6
16
4
Compound 2 under normal conditions of electron
bombardment showed no molecular ion. However,
using an electrospray instrument it exhibited a mole-
cular ion at m/z 300. This corresponds to the proposed
1
molecular formula of C H NO The H NMR and
1
7
18
4
13C NMR (Table 4) for ring A, B, D closely resemble
those reported for the anhydrolycorinium ion (Petitt et
1
al., 1984). The H NMR showed four singlets at 9.36,
.77, 7.63 and 6.38 assigned to H-7, H-11, H-8 and the
7
3.1. Plant material
methylenedioxy, respectively. It also showed multiplets
at 6.45, 5.31, 5.00, 4.61 and 3.32 assigned to H-3, H-11b,
H-5, H-2 and H-4, respectively. The two ddd at 5.49 and
Crinum moorei (Hook f.) plants were obtained in
December 1997 from Green Goblin Nursery, Durban,
South Africa and their identity con®rmed by Dr. T.J.
Edwards, School of Botany and Zoology, University of
Natal Pietermaritzburg. A voucher specimen (Elgora-
shi2 NU) was deposited in the University of Natal
Herbarium, Pietermaritzburg.
5
.48 were assigned to the two protons at position 1. The
multiplicities of the peaks relating to the aliphatic por-
tion of the molecule were completely assigned by COSY
analysis. It showed a correlation between H-2 (ꢀ 4.61)
and the one proton multiplet of H-3 at ꢀ 6.41 and the
ddd of H-1ꢁ and H-1ꢂ at 5.49 and 5.48. In addition,
there was a correlation between the ddd of both protons
at position 1 and that of H-11b at ꢀ 5.31. The assign-
ment of the signal at 5.31 to H-11b was supported by
the long range coupling between the signal at 5.31 and
that of H-11 (7.77). The deshielding of C-5 and C-4
protons is due to their a- and b-positions with respect to
the nitrogen of the salt (Bastida et al., 1992).
3.2. Extraction and isolation
The dried and powdered non ¯owering whole plants
ꢀ
(62.5 g) were extracted with Petr. ether (60±80 ) and
95% EtOH for 40 h each using a Soxhlet apparatus.
The ethanolic extract was evaporated under reduced
pressure and the residue was treated with 4% aq. HOAc.
The aqueous acidic solution was ®ltered. The solution
was basi®ed with NH OH to pH 9.5 after removal of
4
neutral material with Et O. The basi®ed solution was
2
The 13C NMR showed four methine sp carbons in
the aromatic region at 142, 127, 105 and 101 ppm
assigned to C-7, C-3, C-8, and C-11. The chemical shifts of
the six quaternary carbons C-3a, C-7a, C-9, C-10, C-11a
and C-11c are shown in Table 4. The placement of the
methoxy group on C-11c was based on the chemical
shift of this carbon atom and was in good agreement
with predictions for 13C chemical shifts derived from a
commercially available modelling program. Lack of
material and marked instability prevented a more
2
extracted with Et O, EtOAc, and n-BuOH to give frac-
2
tion A, B, C, respectively (Ghosal et al., 1983).
Fractions A and B were combined and kept overnight
in MeOH at room temp. to give lycorine as a powder
(81 mg). The remaining crude extract subjected to VLC
on silica gel eluting with CHCl and then with CHCl
3
3
enriched gradually with MeOH up to 50% MeOH to
give ®ve fractions. Fraction I was developed on PLC (2
mm) using CHCl ±MeOH (9:1) which were developed
3
again using CHCl ±Et NH (20:1), CHCl ±Et NH (40:1),
3
2
3
2
benzene±MeOH (9:1) to give epibuphanisine (56 mg), 1-
O-acetyllycorine (32 mg), undulatine (11 mg) and 3-O-
acetylcrinine (8 mg).
Fraction II was developed on TLC using CHCl3±
Et NH (20:1) to give two bands which were developed
2
further on CHCl ±Et NH (40:1) twice to give epivitta-
3
2
tine (53 mg), cherylline (35 mg) and CHCl ±MeOH
3
(10:1) to give crinamidine (7 mg). Fractions III, IV, V
were developed on PLC using CHCl ±Et NH (20:1) to
3
2

640
E.E. Elgorashi et al. / Phytochemistry 56 (2001) 637±640
give powelline (20 mg), crinine (36 mg), and 1-epidea-
cetylbowdenisine (16 mg).
Fraction C was subjected to VLC using silica gel
Acknowledgements
E.E.E. acknowledges a PhD scholarship awarded by
DAAD. S.E.D. and J.V.S. thank the University of
Natal Research Fund and the National Science Foun-
dation, Pretoria, for ®nancial support. NMR spectra
were recorded by Mr. M. Watson and Mr. J. Rayn,
School of Chemical and Physical Sciences, University of
Natal Pietermaritzburg.
eluting with CHCl enriched gradually with MeOH to
3
give two fractions. Fraction I was developed on TLC
using CHCl ±MeOH (2:1) to give compound 2 (7 mg).
3
Fraction II was developed on PLC (2 mm) using CHCl3±
CH Cl ±EtOH±MeOH (7:7:7:4) and NH vapour to
give compound 1 (11 mg).
2
2
3
3
.2.1. Conversion of tyramine to its acetate
Twenty-nine mg of tyramine were heated with equal
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0
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0
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0
7.5, 2H-8 ), 2.81 t (7.5, 2H-7 ), 2Me (s, 2.02 and 1.93).
3
C NMR (CD OD): 173.2 s and 171.4 s (2 OCOCH ),
3
3
0
0
0
0
0
1
1
50.7 s (C-1 ), 130.7 d (C-2 and C-6 ), 138.9 s (C-4 ),
0
1
13
1
tochemistry 22, 581±584.
983. H and C analysis of lycorine and a-dihydrolycorine. Phy-
0
0
22.6 d (C-3 and C-5 ), 41.9 t (C-8 ), 35.7 t (C-7 ), 22.5
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3
.2.2. Tyramine
1
0
0
H NMR (CD OD): 7.11 d (8.42, H-2 and H-6 ), 6.79
3
0
0
0
d (8.42, H-3 and H-5 ), 3.13 t (7.14, 2H-8 ), 2.88 t (7.09,
1
984. Alkaloidal constituents of Crinum latifolium and Crinum bul-
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0 0
H-7 ). C NMR (CD OD): 157.2 s (C-1 ), 130.7 d (C-
3
13
2
2
0
0
0
0
0
3
and C-6 ), 130.9 s (C-4 ), 116.4 d (C-3 and C-5 ), 43.4t
0
0
Petitt, G.R., Gaddamidi, V., Goswami, A., Cragg, G.M., 1984. Anti-
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(
C-8 ), 36.7 t (C-7 )
3
.2.3. Compound 1
Amorphous compound (11 mg)[ꢂ] +53.3 (CHCl ,
È
È È
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2
9
ꢀ
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3
1
13
c, 0.045) H and CNMR (see Tables 1 and 2). GC±MS
7
1
0 eV, m/z (% rel. int.): 286 (<1%) 258 (100), 248 (26),
86 (18), 129 (9), 115 (14), 107 (11), 44 (35), 43 (56), 42
1
1, 27±52.
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1
1
3
.2.4. Compound 2
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1
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+
Table 4), electrospray (% rel. int.): 300 [M , 5), 292 (5),
2
84 (100), 266 (5).