Chem. Pharm. Bull.

64, 793–799 (2016)

Vol. 64, No. 7793

Special Collection of Papers

This article is dedicated to Professor Satoshi Ōmura in celebration of his 2015 Nobel Prize.

Regular Article

New Lycopodine-Type Alkaloids from Lycopodium carinatum

Noriyuki Kogure,a Moe Maruyama,a Sumphan Wongseripipatana,b
Mariko Kitajima,a and Hiromitsu Takayama*,a
a
 Graduate School of Pharmaceutical Sciences, Chiba University; 1–8–1 Inohana, Chuo-ku, Chiba
260–8675, Japan: and b Faculty of Pharmaceutical Sciences, Chulalongkorn University;
Bangkok 10500, Thailand.
Received February 22, 2016; accepted March 16, 2016; advance publication released online March 25, 2016

The structures of new lycopodine-type alkaloids, lycopocarinamines A–F, which were isolated from
Lycopodium carinatum, were elucidated by spectroscopic analysis and chemical conversions. The proposed
structure of lycocarinatine A was revised.
Key words Lycopodium alkaloid; lycopocarinamine; structure elucidation; chemical conversion

The genus Lycopodium, which belongs to Lycopodiaceae,
consists of more than 200 species from which many kinds of
unique alkaloids, the so-called Lycopodium alkaloids, have
been isolated.1–4) Among the Lycopodium alkaloids, huper-
zine A isolated from L. serratum in 1986 has been shown to
have acetylcholinesterase (AChE) inhibitory activity (IC50:
0.082 µM) and to improve memory disorders in Alzheimer’s
disease patients.5,6) It was reported that some Lycopodium al-
kaloids possessing skeletons different from huperzine A were
able to enhance nerve growth factor (NGF) mRNA expression
and production in human glial cells,7,8) had anti-human immu-
nodeficiency virus (HIV)-1 activity,9,10) or inhibited lipopoly-
saccharide (LPS)-induced pro-inflammatory factors in BV2
microglia and macrophages.11) Because of their useful bio-
logical activities, Lycopodium alkaloids are attractive research
targets in natural product chemistry, synthetic chemistry, and
medicinal chemistry.12–14)
In the course of our research on bioactive Lycopodium al-
kaloids, we have isolated novel Lycopodium alkaloids and de-
termined their structures by spectroscopic analyses and total
syntheses.15–26)
Lycopodium alkaloids are classified into four groups on the
basis of their structures, i.e., fawcettimine, lycodine, lycopo-
dine, and miscellaneous groups. In this paper, we describe the
structure elucidation of six new Lycopodium alkaloids having
the lycopodine-type skeleton, which were isolated from L.
carinatum,27) and the structure revision of reported alkaloid
lycocarinatine A.28)
Results and Discussion
Lycopodium carinatum purchased in Bangkok, Thailand,
was dried, powdered, and extracted with MeOH. The crude
alkaloidal fraction obtained by a conventional procedure from
the MeOH extract was successively purified by SiO2 and
amino-silica gel column chromatography to give 26 alkaloids,
six of which were new alkaloids (1, 2, 5, 7, 9, 10).
The molecular formula of new alkaloid 1, named lycopo-
 To whom correspondence should be addressed.  e-mail: takayamah@faculty.chiba-u.jp
* 
carinamine A, was established as C26H37 NO6 from high res-
olution-electrospray ionization (HR-ESI)-MS [m/z 460.26722
(MH+)], which possessed ten carbons more than common
Lycopodium alkaloids (Fig. 1). 1H- and 13C-NMR data (Table
1) showed signals characteristic of common Lycopodium al-
kaloids, namely, a secondary methyl group [δ H 0.88 (3H, d),
δC 23.4], two sets of aminomethylenes [δ H 3.24 (td, H-1), δ H
3.20 (td, H-9), δ H 2.47 (2H, overlapped, H-1 and 9), δC 46.6,
δC 45.6], and two oxymethines [δ H 4.70 (s, H-6), δ H 3.47 (d,
H-5), δC 82.2, δC 71.5]. Furthermore, the signals of 1,2,4-tri-
substituted aromatic protons [δ 6.85 (d, J=7.7 Hz), δ 6.70 (d,
J=2.1 Hz) and δ 6.69 (dd, J=7.7, 2.1 Hz)], a methoxy group [δ
3.88 (3H, s)], and two adjacent methylene protons [δ 2.90 (2H),
δ 2.61 (2H)] implied the presence of a dihydroferulic acid unit.
1
H–1H correlation spectroscopy (COSY) and 1H-detect-
ed heteronuclear multiple quantum coherence (HMQC)
analyses (Fig. 1) indicated the presence of three carbon
chains (a: –CH2CH2CH2CHCH–; b: –CH2CH2CH2–; c:
–CHCHCH2CHMeCH2–), and the gross structure of 1 was
established by heteronuclear multiple bond connectivity
(HMBC) analyses as follows. HMBC cross peaks between
H-5 (δ 3.47) and the carbons of fragment c (δ 44.1, C-7 and δ
82.2, C-6) showed that fragment c was attached to C-5 posi-
tion. The cross peak between H-7 and the terminal carbon of
fragment b (δ 30.9, C-11) and a quaternary carbon (δ 69.7)
indicated that fragment b was attached to C-7 via the qua-
ternary carbon (δ 69.7). HMBC correlations between H-5 and
another quaternary carbon (δ 58.4), and between H-14 (δ 2.24)
and C-4 and the same quaternary carbon (δ 58.4) implied
that C-14 and C-4 were connected via the quaternary carbon
(δ 58.4, C-13). Correlations from H-1 to C-9 and C-13 indi-
cated that C-1, C-9, and C-13 were connected to each other via
a nitrogen atom. HMBC correlation between H-6 (δ 4.70) and
an ester carbonyl carbon showed that the dihydroferulic acid
unit was connected to C-6 position.
The stereochemistry of 1 was elucidated as follows. Nu-
clear Overhauser effect (NOE) correlations of H-4/H-9 and
© 2016 The Pharmaceutical Society of Japan
794 Chem. Pharm. Bull. Vol. 64, No. 7 (2016)

Fig. 1. Lycopocarinamine A (1)

Table 1. NMR Data for 1–3 in CDCl3a)

Lycopocarinamine A (1) Lycopocarinamine B (2) Lycocarinatine A (3, revised)

Position
1 13 1 13 1 13
H C H C H C

1 3.24 (td, 13.7, 3.4) 46.6 3.25 (td, 13.7, 4.1) 46.6 3.55 (td, 13.5, 3.6) 63.3
2.47 (overlapped) 2.57–2.44 (overlapped) 2.84 (m)
2 1.98 (overlapped) 18.9 1.95 (overlapped) 18.8 1.90 (m) 22.2
1.34 (d, 8.2) 1.34 (overlapped) 1.80 (m)
3 1.84 (overlapped) 22.35 1.85 (overlapped) 22.4 1.90 (m) 20.3
1.41 (d, 13.1) 1.42 (br d, 13.1) 1.42 (br d, 14.0)
4 2.33 (br s) 31.1 2.38 (br t, 8.6) 31.1 2.12 (m) 37.2
5 3.47 (d, 6.9) 71.5 3.51 (d, 6.9) 71.6 3.38 (d, 6.8) 70.8
6 4.70 (s) 82.2 4.73 (s) 82.2 4.58 (s) 82.1
7 1.79 (overlapped) 44.1 1.80 (overlapped) 44.2 1.73 (br s) 45.2
8 1.90 (overlapped) 33.4 1.92 (overlapped) 33.4 2.05 (td, 12.9, 5.2) 33.1
1.36 (t, 6.2) 1.36 (overlapped) 1.24 (m)
9 3.20 (td, 13.1, 3.4) 45.6 3.20 (td, 12.4, 3.4) 45.7 4.02 (td, 12.7, 3.3) 59.5
2.47 (overlapped) 2.57–2.44 (overlapped) 2.95 (m)
10 1.98 (overlapped) 20.5 2.00 (overlapped) 20.6 2.84 (m) 16.4
1.50 (d, 13.7) 1.52 (br d, 13.7) 1.52 (br d, 12.4)
11 1.82 (overlapped) 30.9 1.87 (overlapped) 31.0 1.63 (td, 14.0, 4.1) 30.3
1.18 (d, 13.7) 1.21 (br d, 13.7) 1.24 (m)
12 69.7 69.7 71.1
13 58.4 58.3 72.9
14 2.24 (dd, 13.7, 6.2) 35.3 2.25 (dd, 13.1, 6.2) 35.4 2.71 (t, 12.7) 27.5
1.18 (d, 13.7) 1.24 (br d, 11.7) 1.80 (m)
15 2.53 (m) 22.26 2.57–2.44 (overlapped) 22.3 2.58 (m) 22.6
16 0.88 (3H, d, 6.2) 23.4 0.88 (3H, d, 6.2) 23.4 0.94 (3H, d, 6.3) 23.2
17 172.5 172.4 173.0
18 2.61 (2H, m) 36.6 2.63 (2H, m) 36.4 2.64 (2H, m) 36.6
19 2.90 (2H, m) 31.1 2.91 (2H, m) 30.8 2.95 (m) 31.6
2.84 (m)
20 131.9 132.6 131.6
21 6.70 (d, 2.1) 111.0 6.73 (d, 1.4) 111.6 6.71 (d, 1.2) 111.3
22 146.5 148.9 146.6
23 144.1 147.6 144.3
24 6.85 (d, 7.7) 114.3 6.80 (d, 7.6) 111.1 6.84 (d, 7.7) 114.3
25 6.69 (dd, 7.7, 2.1) 120.9 6.74 (dd, 7.6, 1.4) 120.2 6.69 (dd, 7.7, 1.2) 121.2
26 3.88 (3H, s) 55.9 3.88 (3H, s) 55.9 3.89 (3H, s) 56.0
27 3.86 (3H, s) 55.9
a) Measured at 600 and 150 MHz, respectively.

H-4/H-11 indicated that H-4 had an axial orientation and
the stereochemistry of C-12 quaternary carbon was estab-
lished, as shown in Fig. 1. NOE correlations of H-4/H-5 and
H-5/H-6 implied that H-4 and H-5 were in syn orientation.
The coupling pattern of H-6 (s) suggested that the dihedral
angles between H-5 and H-6, and between H-6 and H-7 were
respectively ca. 90°, as depicted in Fig. 1. NOE correlations
of H-6/H-15 demonstrated that the methyl group was in an
equatorial position. From these analyses, the structure of 1
was elucidated as shown in Fig. 1. Lycopocarinamine A (1)
 Chem. Pharm. Bull.
Vol. 64, No. 7 (2016)795
Chart 1. Chemical Conversions of Lycopocarinamine A (1)
Fig. 2. Structures of Lycocarinatine A (Revised and Reported)
is the second example of a lycopodine-type alkaloid having a
5,6,12-tri-oxygenated skeleton.
The molecular formula of new alkaloid 2, named lycopo-
carinamine B, was deduced from HR-ESI-MS experiments
to be C27H39NO6 [m/z 474.28222 (MH+)], which had an extra
CH2 compared to 1. The 1H-NMR data of 2 were very similar
to those of 1 except for the presence of an additional methoxy
signal at δ 3.86 (3H, s). Taking these facts into consideration,
2 was presumed to be an O-methyl derivative of 1. To confirm
the structure inferred by spectroscopic analyses above, the
chemical conversion of 1 was attempted (Chart 1). 1 was treat-
ed with freshly prepared CH2N2 solution to afford 2. 1H-NMR
data of the methylated compound was completely identical
with those of natural 2 and therefore, the structure of 2 was
established as shown.
The molecular formula of alkaloid 3 was established as
C26H37 NO7 from the protonated molecular ion peak [m/z
476.26125 (MH+)] in the HR-ESI-MS spectrum, which had an
extra oxygen atom compared to 1. The 1H- and 13C-NMR data
of 3 resembled those of 1 except for the signals around the ni-
trogen atom (H2-1, 9 and C-1, 9, 13). From these data, alkaloid
3 was deduced to be an N-oxide derivative of 1. To confirm
the structure of 3, a chemical conversion from 1 was executed.
1 was treated with m-chloroperbenzoic acid (mCPBA) in
CH2Cl2 at 0°C to give 3 in a quantitative yield. The spectral
data of the oxidized compound were completely identical with
those of the natural product and therefore, the structure of 3
was confirmed.
We noticed that the spectral data of 3 were in good agree-
ment with those of recently reported lycocarinatine A (4, re-
ported)28) (Fig. 2). In that study, the authors observed HMBC
correlations between H-5 and C-12, and between H-6 and
C-13. However, as those were four-bond correlations, the au-
thors must have erred in the assignment of the signals of [H-5,
C-5] for [H-6, C-6]. We hereby propose the revised structure
of lycocarinatine A as shown in Fig. 2.
The HR-ESI-MS spectrum of new alkaloid 5, named ly-
copocarinamine C, gave a protonated molecular ion peak at
m/z 458.25452 ([MH]+) that corresponded to the molecular
formula C26H35NO6 (Fig. 3). 1H-NMR data (Table 2) showed
signals characteristic of Lycopodium alkaloids, namely, a sec-
ondary methyl group [δ 1.00 (3H, d)], two sets of aminometh-
ylenes [δ 2.93 (m, H-1) and δ 2.53 (m, H-1), δ 2.62 (2H, m,
H2-9)], and an olefinic proton [δ 5.40 (t, H-11)]. Furthermore,
the signals of 1,2,4-trisubstituted aromatic protons [δ 6.68 (dd,
J=8.2, 2.1 Hz), δ 6.82 (d, J=8.2 Hz), and δ 6.68 (d, J=2.1 Hz)],
a methoxy group [δ 3.88 (3H, s)], and two adjacent methylene
protons [δ 2.85 (2H), δ 2.58 (2H)] indicated the presence of a
dihydroferulic acid unit. The 1H- and 13C-NMR data highly
resembled those of known Lycopodium alkaloid malycorin B
(6).29) The only differences between 5 and 6 were the absence
of an acetyl group in 5 and the chemical shift of proton at C-8
(5; δ 3.30, 6; δ 4.46). Judging from these spectroscopic data,
5 was deduced to be an 8-O-deacetyl derivative of malycorin
B (6).
New alkaloid 7, named lycopocarinamine D, was obtained
as a colorless amorphous powder and its molecular formula
was established as C16H25NO3 by HR-ESI-MS [m/z 280.18909
(MH)+] (Fig. 4). 1H-NMR data (Table 3) showed signals
characteristic of common Lycopodium alkaloids, namely, one
796 Chem. Pharm. Bull. Vol. 64, No. 7 (2016)

Fig. 3. Structures of Lycopocarinamine C (5) and Malycorin B (6)

Table 2. NMR Data for 5 and 6 in CDCl3a) The molecular formula of new alkaloid 9, named lyco-
pocarinamine E, was deduced to be C18H27 NO5 from HR-
Lycopocarinamine C (5) Malycorin B (6) ESI-MS [m/z 338.19654 (MH)+]. The 1H- and 13C-NMR data
Position
1
H 13
C 1
H 13
C markedly resembled those of lycopocarinamine D (7) except
for the chemical shifts of H-8 and C-8 and the existence of an
1 2.93 (m) 48.5 2.94 (m) 48.4
acetoxy group [δ Η 2.05 (3H, s), and δC 170.6 and δC 21.2] in 9.
2.53 (m) 2.54 (m)
Judging from these data, 9 was deduced to be an 8-acetoxy
2 1.77 (m) 24.9 1.76 (m) 24.8
derivative of 7. The configuration of the acetoxy group at C-8
1.64 (m) 1.63 (br s)
was shown to be β by the coupling constant between H-8 and
3 1.77 (m) 22.5 1.85 (br s) 22.6
H-15 (J8,15=11.0 Hz). Lycopocarinamine E (9) is the first exam-
1.42 (br d, 8.9) 1.42 (d, 8.8)
4 1.95 (m) 43.9 1.95 (m) 43.7
ple of lycopodine-type alkaloids having three hydroxy groups
5 3.60 (d, 6.2) 71.1 3.62 (br s) 71.4
at C-8, C-11, and C-12 positions.
6 5.13 (s) 75.1 4.96 (m) 75.0 The 1H-NMR spectrum of lycopocarinamine F (10) showed
7 2.53 (m) 51.2 2.67 (m) 47.9 specific signals due to Lycopodium alkaloids, namely, one
8 3.30 (dd, 10.3, 5.5) 79.5 4.46 (dd, 11.2, 5.0) 80.4 secondary methyl group [δ 0.89 (3H, d, H3-16)], two sets of
9 2.62 (2H, m) 45.0 2.67 (2H, m) 45.0 aminomethylenes [δ 2.98 and δ 2.55 (H2-1), δ 2.67 (2H, H2-9)],
10 2.26 (m) 26.2 2.25 (m) 26.4 and an olefinic proton [δ 5.53 (H-11)] (Table 4). The 1H- and
13
2.03 (m) 2.01 (m) C-NMR spectra evidently resembled those of known alka-
11 5.40 (t, 4.1) 118.8 5.49 (t, 4.4) 120.2 loid lycofoline (11)31) except for the existence of signals for an
12 139.5 138.3 acetyl group [δ H 2.09 (3H, s), δC 171.0 and δC 21.4] in 10. The
13 56.0 55.7 molecular formula of 10 was established by HR-ESI-MS [m/z
14 2.21 (dd, 13.1, 6.2) 31.6 2.25 (m) 31.8 306.20928 (MH)+] as C18H27 NO3, which indicated that 10 had
1.05 (m) 1.10 (t, 12.4) an extra C2H2O compared to lycofoline (11). From the data
15 2.72 (m) 31.6 2.94 (m) 28.6 mentioned above, 10 must be an acetyl derivative of lycofoline
16 1.00 (3H, d, 6.2) 20.3 0.88 (3H, d, 6.2) 19.8 (11). The chemical shift of H-8 in 10 was lower than that in
17 172.7 172.2 lycofoline (11), indicating that the acetyl group was attached
18 2.58 (2H, t, 6.9) 36.4 2.57 (2H, t, 8.1) 36.3 to O-8 position (Fig. 5).
19 2.85 (2H, t, 6.9) 30.7 2.85 (2H, td, 7.5, 2.1) 30.7
20 132.1 132.1
Experimental
21 6.68 (d, 2.1) 110.9 6.68 (d, 1.8) 110.8
General 1H- and 13C-NMR spectra: JEOL JNM ECA-600
22 146.4 146.4
or JEOL ECZ-600 or ECP-600 at 600 MHz (1H-NMR)
23 144.0 144.0
and 150 MHz (13C-NMR), respectively. HR-ESI-MS: JEOL
24 6.82 (d, 8.2) 114.3 6.82 (d, 7.9) 114.3
AccuTOF-GCv. UV: JASCO V-560. Optical rotation: JASCO
25 6.68 (dd, 8.2, 2.1) 120.8 6.66 (dd, 7.9, 1.8) 120.8
P-1020. TLC: precoated silica gel 60 F254 plates (Merck,
26 3.88 (3H, s) 55.9 3.88 (3H, s) 55.9
COMe 170.9
0.25 mm thick), NH-TLC (Fuji Silysia Chemical). Flash
COMe 2.12 (3H, s) 21.3
column chromatography: Silica gel 60N (Kanto Chemi-
cal, 40–50 µm). Amino-silica gel column chromatography:
a) Measured at 600 and 150 MHz, respectively.
NH-DM1020 (Fuji Silysia Chemical).
Plant Material Lycopodium carinatum was purchased
secondary methyl group [δ 0.87 (3H, d, H3-16)] and two sets of from a plant market in Bangkok. The plant was identi-
aminomethylenes [δ 3.22 and δ 2.50 (H2-1), δ 3.53 and δ 2.45 fied by Dr. S. Wongseripipatana and a voucher specimen
(H2-9)]. 13C-NMR data indicated the presence of a ketone car- (No.20100201) is stored at the Graduate School of Pharmaceu-
bon [δ 214.8 (C-5)], an oxymethine carbon [δ 74.3 (C-11)], and tical Sciences, Chiba University, Japan.
two quaternary carbons [δ 70.0 (C-12), δ 60.9 (C-13)] attached Extraction and Isolation The dried, powdered whole
to the heteroatom. The 1H- and 13C-NMR data were similar to plant (451.2 g) was extracted with MeOH four times (twice at
those of known alkaloid lycoposerramine N (8)26) except for room temperature, twice under reflux). The extract was evapo-
the absence of an acetyl group in 7. From the data mentioned rated to give a crude MeOH extract (150.2 g). The MeOH
above, 7 was deduced to be a deacetyl derivative of lycoposer- extract was dissolved in 2% aq. tartaric acid (2.0 L) and the
ramine N (8). mixture was filtered to remove the insoluble part. The filtrate
 Chem. Pharm. Bull.
Vol. 64, No. 7 (2016)797

Fig. 4. Structures of 7–9

Table 3. NMR Data for 7–9 in CDCl3a)

Lycopocarinamine D (7) Lycopocarinamine E (9) Lycoposerramine N (8)

Position
1 13 1 13 1 13
H C H C H C

1 3.22 (td, 13.8, 3.4) 46.6 3.16 (td, 14.4, 4.1) 47.0 3.22 (td, 14.2, 3.8) 46.6
2.50 (overlapped) 2.54 (overlapped) 2.49 (dd, 14.7, 5.2)
2 1.90 (m) 18.0 1.90 (m) 17.8 1.90 (qt, 13.7, 5.4) 18.0
1.37 (br d, 13.8) 1.38 (overlapped) 1.40 (br d, 15.0)
3 2.05 (m) 19.4 2.04 (br d, 11.0) 19.0 2.10 (br d, 12.5) 19.7
1.67–1.57 (overlapped) 1.69–1.50 (overlapped) 1.66 (m)
4 3.72 (m) 42.5 3.76 (dd, 12.4, 4.1) 42.4 3.44 (dd, 11.9, 3.7) 42.4
5 214.8 213.7 213.4
6 3.48 (dd, 16.5, 6.9) 45.6 3.21 (dd, 17.2, 6.9) 42.5 2.85 (dd, 17.2, 6.9) 45.2
2.28 (d, 16.5) 2.44–2.40 (overlapped) 2.32 (dd, 17.4, 1.2)
7 2.26 (overlapped) 39.7 2.44–2.40 (overlapped) 38.6 2.20 (t, 3.5) 38.9
8 1.96 (td, 13.1, 3.4) 37.3 5.24 (dd, 11.0, 4.1) 74.1 2.00 (td, 13.1, 4.0) 37.3
1.32 (br d, 11.0) 1.31 (br d, 11.9)
9 3.53 (br t, 13.1) 41.4 3.51 (td, 14.4, 5.5) 41.3 3.33 (td, 12.8, 3.1) 41.9
2.45 (m) 2.57–2.51 (overlapped) 2.46 (m)
10 2.50 (overlapped) 31.4 2.57–2.51 (overlapped) 31.9 2.42 (m) 27.1
1.67–1.57 (overlapped) 1.69–1.50 (overlapped) 1.75 (dq, 15.0, 2.5)
11 4.01 (s) 74.3 4.05 (d. 1.4) 71.5 4.98 (t, 2.7) 75.2
12 70.0 64.3 69.7
13 60.9 61.0 60.7
14 2.34 (dd, 13.8, 4.8) 37.0 2.44–2.40 (overlapped) 35.8 2.38 (m) 37.1
1.26 (overlapped) 1.40–1.36 (overlapped) 1.26 (t, 13.1)
15 1.51 (m) 24.1 1.69–1.50 (overlapped) 30.9 1.48 (m) 24.1
16 0.87 (3H, d, 6.2) 22.4 0.88 (3H, d, 6.2) 22.7 0.87 (3H, d, 6.1) 22.4
COMe 170.6 169.5
COMe 2.05 (3H, s) 21.2 2.07 (3H, s) 21.9
a) Measured at 600 and 150 MHz, respectively.

was extracted with petroleum ether (1.0 L) and the acidic aq.
layer was basified with Na2CO3 (pH 10) and extracted with
5% MeOH/CHCl3 (1.75 L×4). The organic layer was dried
over MgSO4, filtered, and evaporated to give the crude alka-
loidal fraction (2.99 g).
The crude alkaloidal fraction was separated by SiO2 flash
chromatography using MeOH/CHCl3 gradient (3–50%),
MeOH, and 10% aq. ammonia/MeOH to give 13 fractions (frs.
A–M). The MeOH eluate (fr. J) was purified by amino-silica
gel open column chromatography with 5% MeOH/AcOEt,
MeOH, and 10% aq. ammonia/MeOH to give six fractions
(frs. J1–J6). The 5% MeOH/AcOEt eluate (fr. J4) was purified
successively by amino-silica gel open column chromatogra-
phy (10–20% EtOH/n-hexane, 2% MeOH/CHCl3) to afford
lycopocarinamine A (4.2 mg), lycopocarinamine B (1.7 mg), ly-
copocarinamine C (1.8 mg), lycopocarinamine D (0.9 mg), and
lycopocarinamine E (0.3 mg). Lycopocarinamine F (3.3 mg)
was isolated from fr. J2 by SiO2 flash column chromatography
(3% MeOH/CHCl3 saturated with ammonia).
Other isolated compounds from the crude alkaloidal frac-
tion were lycopodine,31) lycocarinatine A,28) 12-epi-lycodoline
N-oxide,32) lycofoline,30) anhydrolycodoline,30) obscurumine
C,33) gnidioidine,30) malycorin B,29) fawcettimine,34) lyco-
poserramine-C,35) lycoposerramine-D,35) lycoposerramine-P,35)
phlegmariurine A,22) phlegmariurine B,36) huperzine A,5) hu-
perzine C,37) 13N-formyl huperzine A,38) lycodoline,32,39) and
798 Chem. Pharm. Bull. Vol. 64, No. 7 (2016)

Table 4. NMR Data for 10 and 11 in CDCl3a) (MH)+ (Calcd for C16H26NO3: 280.19127)
Lycopocarinamine E (9)
Lycopocarinamine F (10) Lycofoline (11)
Position Colorless amorphous powder; 1H- and 13C-NMR, see Table
1
H 13
C 1
H 13
C 3; [α]D25 +44.8 (c=0.008, MeOH); HR-ESI-MS m/z 338.19654
1 2.98 (t-like, 9.6) 48.5 2.96 (m) 48.6
(MH)+ (Calcd for C18H28NO5: 338.19675)
2.55 (d, 11.0) 2.54 (m)
Lycopocarinamine F (10)
2 1.60 (2H, m) 24.7 1.77 (m) 24.9 Colorless amorphous powder; 1H- and 13C-NMR, see Table
1.62 (m) 4; [α]D25 −3.4 (c=0.14, MeOH); HR-ESI-MS m/z 306.20928
3 1.78 (m) 23.2 1.77 (m) 23.1 (MH)+ (Calcd for C18H28NO3: 306.20692)
1.51 (m) 1.48 (m) Chemical Conversion of Lycopocarinamine A (1) into
4 2.05–1.88 (overlapped) 47.1 1.94 (m) 47.6 Lycopocarinamine B (2) To a solution of lycopocarinamine
5 3.97 (t, 4.8) 67.8 3.96 (t, 5.5) 68.0 A (1, 1.0 mg) in MeOH (0.5 mL) was added freshly prepared
6 2.05–1.88 (2H, overlapped) 33.0 2.12 (d, 15.8) 32.6 ether solution of CH2N2 (excess). The mixture was stirred for
1.89 (dt, 15.8, 6.2) 1 h at room temperature. After the decomposition of CH2N2
7 2.70 (t, 4.8) 42.5 2.54 (m) 45.8 (the yellowish color faded) was completed, the solvent was
8 4.44 (dd, 10.2, 4.8) 81.5 3.23 (br s) 79.9 removed under reduced pressure. The residue was purified by
9 2.67 (2H, m) 45.1 2.66 (2H, m) 45.1 SiO2 flash chromatography (MeOH/CHCl3 gradient) to give
10 2.22 (m) 26.3 2.24 (m) 26.2 semi-synthetic 2 (0.37 mg, 36%). All the spectroscopic data of
2.05–1.88 (overlapped) 2.01 (m) semi-synthetic 2 were identical with those of natural 2.
11 5.53 (t, 4.1) 117.9 5.45 (t, 3.7) 116.4 Chemical Conversion of Lycopocarinamine A (1) into
12 142.1 143.5 Lycocarinatine A (3) To a stirred solution of 1 (0.5 mg) in
13 56.1 56.4 dry CH2Cl2 (0.1 mL) was added mCPBA (77%, 0.27 mg) at
14 2.28 (dd, 13.7, 6.8) 32.5 2.22 (dd, 13.0, 6.8) 32.4 0°C under argon atmosphere. After the reaction mixture was
1.10 (t, 13.0) 1.06 (t, 13.0)
stirred at 0°C for 30 min, it was directly subjected to amino-
15 3.07 (m) 28.1 2.77 (m) 31.5
silica gel open column chromatography (MeOH/AcOEt gradi-
16 0.89 (3H, d, 6.2) 19.8 1.02 (3H, d, 6.9) 20.4
ent) to give semi-synthetic 3 (0.67 mg, quant.). All the spec-
COMe 171.0
troscopic data of semi-synthetic 3 were identical with those of
COMe 2.09 (3H, s) 21.4
natural 3.
a) Measured at 600 and 150 MHz, respectively.

Acknowledgments This work was supported by JSPS

KAKENHI Grant Numbers 25293023, 26293023 and
25460005.

Conflict of Interest The authors declare no conflict of

interest.

Fig. 5. Structures of Lycopocarinamine F (10) and Lycofoline (11)
lycodine.18)
Lycopocarinamine A (1)
Colorless amorphous powder; 1H- and 13C-NMR, see Table
1; [α]D27 −8.9 (c=0.14, MeOH); UV λmax (MeOH) 282.5, 229.5,
216.0 nm; HR-ESI-MS m/z 460.26722 (MH)+ (Calcd for
C26H38NO6: 460.26991)
Lycopocarinamine B (2)
Colorless amorphous powder; 1H- and 13C-NMR, see Table
1; [α]D27 −10.8 (c=0.11, MeOH); UV λmax (MeOH) 279.5,
229.5, 206.0 nm; HR-ESI-MS m/z 474.28222 (MH)+ (Calcd for
C27H40NO6: 474.28556)
Lycopocarinamine C (5)
Colorless amorphous powder; 1H- and 13C-NMR, see Table
2; [α]D26 −4.0 (c=0.08, MeOH); UV λmax (MeOH) 281.5,
230.0, 215.5 nm; HR-ESI-MS m/z 458.25452 (MH)+ (Calcd for
C26H36NO6: 458.25426)
Lycopocarinamine D (7)
Colorless amorphous powder; 1H- and 13C-NMR, see Table
3; [α]D24 −9.5 (c=0.04, MeOH); HR-ESI-MS m/z 280.18909
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