Synthesis and structure-activity relationship studies of cytotoxic anhydrovinblastine amide derivatives

Article abstract of DOI:10.1021/np900157t

A series of 3-demethoxycarbonyl-3-amide methyl anhydrovinblastine derivatives (5b-24b) was designed, synthesized, and evaluated for their proliferation inhibition activities against two tumor cell lines (A549 and HeLa). Most of the amide anhydrovinblastin

Full text of DOI:10.1021/np900157t

1170
J. Nat. Prod. 2009, 72, 1170–1177
Synthesis and Structure-Activity Relationship Studies of Cytotoxic Anhydrovinblastine Amide
Derivatives
Yong Shao,†,‡ Han-Kun Zhang,†,‡ Hong Ding,§ Hai-Tian Quan,§ Li-Guang Lou,§ and Li-Hong Hu*,†
Shanghai Research Center for the Modernization of Traditional Chinese Medicine, Shanghai Institute of Materia Medica, Chinese Academy of
Sciences, Shanghai 201203, People’s Republic of China, and Department of Molecular Pharmacology, Shanghai Institute of Materia Medica,
Chinese Academy of Sciences, Shanghai 201203, People’s Republic of China
ReceiVed March 17, 2009
A series of 3-demethoxycarbonyl-3-amide methyl anhydrovinblastine derivatives (5b-24b) was designed, synthesized,
and evaluated for their proliferation inhibition activities against two tumor cell lines (A549 and HeLa). Most of the
amide anhydrovinblastine derivatives exhibited potent cytotoxicity, with the size of the introduced substituents being
the foremost factor in determining the resultant cytotoxic activity. Test results in vivo against sarcoma 180 of three
potent compounds (6b, 12b, and 24b) indicated that the introduction of an amide group at the 22-position of
anhydrovinblastine (1e) improved both potency and toxicity.
Vinblastine (1a) and vincristine (1b), dimeric indole alkaloids
isolated from Catharanthus roseus (L.) G. Don (Apocynaceae),1 have
been used widely as clinically important antitumor agents in cancer
chemotherapy for more than 40 years.2 These two alkaloids, although
structurally almost identical, differ markedly in the type of tumors that
they affect and in their toxic properties.3 From the 1970s, extensive
chemical research has been undertaken in an effort to yield more active
and less toxic analogues exhibiting a wider spectrum of anticancer
efficacy. Most of the research had mainly focused on the semisynthesis
or total synthesis of vinblastine analogues from the two parts of the
dimeric structure, the vindoline and velbenamine portions, by carbon
skeleton modification and functional group transformation.4 To date,
these efforts have led to only two approved drugs, vindesine (1c)5
and vinorelbine (1d).6 Therefore, new ideas and approaches are needed
to extend investigations of the use of these alkaloids as anticancer
agents. Anhydrovinblastine (1e), a synthetic precursor of vinorelbine,
is currently in phase II clinical trials.7 Compared with 1b and 1d,
anhydrovinblastine has shown superior antitumor activity to human
tumor xenografts of non-small-cell lung cancer and cervical cancer at
equitoxic doses.8 Although having poor cytotoxicity in vitro, 1e shows
advantages in terms of efficacy, tolerability, and range of activity over
vinorelbine (1d) in vivo. These successful developments inspired us
to design new derivatives for searching novel promising dimeric indole
alkaloid antitumor agents with better therapeutic indices.
is sensitive to the overall biological properties, including the
potency, toxicity, and tumor spectrum.9-12 In particular, vindesine
(1c) differs slightly from vinblastine (1a) by having an amide group
instead of an ester group at the C-22 position and possesses an
experimental antitumor spectrum that resembles that of vincristine
(1b) rather than that of the parent alkaloid vinblastine, while its
toxicological profile has suggested a potential for reduced neuro-
toxicity relative to that of 1b.13 Additionally, there has been a lack
of research in directly exploiting the C-22 position of the lead
compound, 1e. Hence, we are interested in exploring the effect of
different amino groups at the C-22 position of anhydrovinblastine
(1e), on the resultant antitumor acivity. In our earlier publications
a variety of carbamates and ether and ester anhydrovinblastine
analogues were prepared for antitumor activity evaluation, and a
candidate was obtained for preclinical trials.14,15 Herein, a total of
20 anhydrovinblastine derivatives (5b-24b) with different amide
groups were designed for biological evaluation in vitro and in vivo,
in order to gain a better understanding of the SAR of nitrogen-
linked analogues at C-22 of compound 1e.
Results and Discussion
The synthesis of targeted amide anhydrovinblastine analogues
was achieved using vindoline as starting material, following the
route as shown in Scheme 1. The key intermediate amine 4 was
prepared easily in four steps from vindoline according to a
previously reported procedure.14 N-Acylation of amine 4 was
obtained with sodium hydride and complex BtCOR, which was
prepared using a different acyl chloride or acid anhydride reacted
with benzotriazole. Subsequently, the N-acylated compounds were
further acetylated at C-4 OH to afford the amide vindoline
derivatives 5a-24a, which, upon coupling with catharanthine (3)
by the modified Polonovski-Potier reaction, produced targeted
amide anhydrovinblastine analogues (5b-24b).16
All the amide compounds (5b-24b) were evaluated in vitro for
their cytotoxicity against human non-small-cell lung cancer (A549)
and cervical epithelial adenocarcinoma (HeLa) cell lines using a
sulforhodamine B (SRB) assay,17 employing 1d and 1e as the
standard for comparison purposes. The IC50 values for the inhibition
of proliferation of the A549 and HeLa cell lines are shown in Table
1. In the aliphatic-substituted series, the acetamide compound 5b
showed comparable cytotoxicity against the A549 cell line to the
positive control 1e (IC50 22 nM for 5b and 30 nM for 1e). However,
the activity of compound 5b against the HeLa cell line decreased
16-fold when compared with 1e (IC50 444 nM for 5b vs 27 nM for
1e). The propionamide derivative 6b (IC50 43 and 57 nM) displayed
only slightly lower activity than 1e against both cell lines. With
Structure-activity relationhip studies on the vindoline portion
of 1a have demonstrated that the substitution at the C-22 position
* To whom correspondence should be addressed. Tel: 86-21-50272221.
Fax: 86-21-50272221. E-mail: simmhulh@mail.shcnc.ac.cn.
† Shanghai Research Center for the Modernization of Traditional Chinese
Medicine.
§ Shanghai Institute of Materia Medica.
‡ These two authors contributed equally to this work.
10.1021/np900157t CCC: $40.75  2009 American Chemical Society and American Society of Pharmacognosy
Published on Web 06/05/2009

Cytotoxic AnhydroVinblastine Amide DeriVatiVes
Journal of Natural Products, 2009, Vol. 72, No. 6 1171
Scheme 1a
a Reagents and conditions: (a) NaH, THF, 0 °C to rt, 6 h; (b) Ac2O, pyridine, 12 h; (c) (i) FeCl3, HCl-H2O buffer (pH ) 2), 8 h; (ii) NaBH4, NH4OH, 0 °C, 15
min; (d) Et3N, DCM, 0 °C to rt, 3 h.
the increasing size of alkyl groups by one additional carbon (7b,
8b, 9b, 10b, and 11b), a significant loss of cytotoxicity against the
two cell lines used was observed. This indicated that small alkyl
groups are tolerated to maintain the cytotoxicity. In addition, the
cytotoxicity data of compounds 9b, 10b, and 11b showed that the
size of groups at the ꢀ-position of the ketone has no effect on
activity. In particular, a cyclopropane (12b) and a cyclobutane
carboxamide (13b) also demonstrated an equally potent cytotoxic
activity to 1e against the A549 cell line (IC50 26 vs 34 vs 30 nM,
respectively), whereas compound 13b displayed about 2-fold lower
activity against the HeLa cell line than 12b and 1e (IC50 68 vs 35
vs 27 nM, respectively).
resulting in undue toxicity, in addition to the single-dose schedules,
a multiple-dose schedule was used. The results of two schedules
of administration both indicated that these compounds showed
remarkable in vivo antitumor activities when compared with the
positive controls, 1d and 1e. The effects of single-dose administra-
tion (20 mg/kg) of these three compounds (6b, 12b, 24b) produced
similar efficacy, but some mice died after day 1 because of the
toxicity. However, the intermittent treatments (days 1, 4, 10 mg/
kg) greatly increased the efficacy with tolerable toxicity (6b, 12b).
Threrfore, the amide group at C-22 improved compound antitumor
activity in vivo. These amide anhydrovinblastine derivatives might
be promising leads for the development of new antitumor agents.
Among 11 aromatic-substituted amide analogues, the phenyl
amide 14b (IC50 27, 26 nM) showed equally potent cytotoxic
activity to 1e, whereas a benzyl-substituted derivative (23b) had
less potent activity (IC50 39, 78 nM) (Table 1). This showed that a
phenyl group was more suitable than a benzyl group to maintain
such activity. Introduction of electron-withdrawing (F, Cl, and NO2)
or electron-donating (OMe) groups at the 4-position of the phenyl
group afforded compounds 15b, 18b, 20b, and 22b, with no
significant effects on cytotoxic activity against the A549 cell line.
The derivatives with electron-withdrawing groups on the phenyl
substituent exhibited better cytotoxic activities against the HeLa
cell line than the compound with an electron-donating group. A
2-methoxy phenyl amide (21b) and a 3,4-methylenedioxy phenyl
amide (24b) showed approximately 3-fold better activity against
the A549 cell line than 1e (IC50 10 vs 8 vs 30 nM), comparable to
1d (IC50 9 nM). No distinct effect on cytotoxic activity against the
A549 cell line was observed due to the positions of Cl in the phenyl
ring (IC50 24 nM for 16b vs 26 nM for 17b vs 25 nM for 18b). A
4-pyridine carboxylic amide (19b, IC50 27, 24 nM) had similar
activity against both cell lines to 4-nitrophenyl (20b, IC50 33, 23
nM).
Experimental Section
General Experimental Procedures. Optical rotations were deter-
mined on a Perkin-Elmer 341 polarimeter. IR spectra were recorded
on a Perkin-Elmer 577 spectrometer. NMR spectra were recorded on
a Varian Mercury-VX300 Fourier transform spectrometer or a Bruker
AM-400 spectrometer. The chemical shifts are reported (in ppm) using
the δ 7.26 signal of CDCl3 (1H NMR) and the δ 77.23 signal of CDCl3
(13C NMR) as internal standards. EIMS were obtained on a Shimadzu
GCMS-QP5050A spectrometer. ESIMS were run on a Bruker Esquire
3000 plus spectrometer in MeOH. Thin-layer chromatographic (TLC)
plates (silica gel 60 GF, with glass support) from Yantai Jiangyou
Company were used for monitoring the progress of a reaction and
visualized with 254 nm UV light and/or sprayed with a 10% solution
of ceric ammonium sulfate (CAS) in phosphoric acid. Unless otherwise
mentioned, all chemicals and materials were used as received from
commercial suppliers without further purification. Tetrahydrofuran was
distilled from sodium/benzophenone under nitrogen. Dichloromethane
was distilled from calcium hydride. Compounds 1d and 1e were
purchased from Shanghai Kang’ai Biological Products Company, Ltd.
Catharanthine tartrate (3) was purchased from Shanghai AnTiKang-
Sheng Plant Chemistry Company, Ltd.
3-Demethoxycarbonyl-3-aminomethyl-4-deacetylvindoline (4). Com-
pound 4 was obtained using the method in a previous report14 in four
steps: [R]20D +38 (c 0.09, CHCl3); 1H NMR (400 MHz, CDCl3) δ 8.64
(1H, s), 6.87 (1H, d, J ) 8.0 Hz), 6.28 (1H, dd, J ) 8.0, 2.4 Hz), 6.06
(1H, d, J ) 2.4 Hz), 5.87-5.79 (2H, m), 3.78 (4H, s), 3.45 (1H, dd, J
) 15.6, 4.4 Hz), 3.36-3.32 (1H, m), 3.24 (1H, s), 3.20 (1H, d, J )
11.6 Hz), 3.16 (1H, d, J ) 11.6 Hz), 2.92 (3H, s), 2.83 (1H, dt, J )
In an effort to further evaluate the antitumor activity of the novel
amide anhydrovinblastine derivatives, three potent cytotoxic com-
pounds, 6b, 12b, and 24b, were selected for testing of their efficacy
in the murine sarcoma 180 model (Table 2). For evaluation of the
influence of the schedule of administration of these amide analogues
on their activities and on the dose that could be injected without

1172 Journal of Natural Products, 2009, Vol. 72, No. 6
Shao et al.
by column chromatography (silica gel; hexane-acetone, 3:1) to give
Table 1. Cytotoxic Activity of Anhydrovinblastine Amide
1
Analoguesa
the desired compound 5a (417 mg, 89%) as a white solid: H NMR
(300 MHz, CDCl3) δ 6.81 (1H, d, J ) 8.1 Hz), 6.24 (1H, d, J ) 8.1
Hz), 6.17 (1H, d, J ) 7.2 Hz), 6.08 (1H, s), 5.82 (1H, dd, J ) 10.2,
4.5 Hz), 5.35 (1H, d, J ) 10.2 Hz), 4.92 (1H, s), 3.78 (3H, s), 3.65
(2H, m), 3.41 (2H, m), 3.33 (1H, s), 3.28 (1H, m), 2.98 (1H, d, J )
13.2 Hz), 2.79 (3H, s), 2.73 (1H, d, J ) 4.8 Hz), 2.59 (1H, s), 2.47
(1H, m), 2.16 (2H, m), 2.09 (1H, s), 2.03 (3H, s), 1.92 (3H, s), 1.22
(1H, m), 0.94 (1H, m), 0.45 (3H, t, J ) 7.2 Hz); 13C NMR (75 MHz,
CDCl3) δ 170.4, 170.3, 161.0, 154.3, 130.3, 125.5, 124.1, 122.6, 105.1,
96.6, 82.1, 77.0, 75.8, 67.3, 55.3, 52.2, 51.5, 50.8, 44.6, 43.7, 42.8,
40.6, 31.2, 23.3, 20.9, 7.5; ESIMS m/z 470.3 [M + 1]+.
General Procedure for the Preparation of Compounds 6a-24a.
To a solution of benzotriazole (238 mg, 2.0 mmol) in CH2Cl2 (10 mL)
was added triethylamine (0.32 mL, 2.2 mmol). Then, a different acyl
chloride (2.2 mmol) in CH2Cl2 (2 mL) was added dropwise at 0 °C.
After being stirred at room temperature for 2 h, the mixture was diluted
with water (50 mL) and extracted with CH2Cl2 (3 × 15 mL). The
organic phase was combined, dried with anhydrous Na2SO4, and
concentrated under vacuum pressure to give an intermediate N-acyl
benzotriazole (BtCOR). Then, to a solution of BtCOR (1.1 mmol) and
compound 4 (386 mg, 1 mmol) in THF (10 mL) was added NaH (60%
in oil, 44 mg, 1.1 mmol). The mixture was stirred for 3 h at room
temperature and quenched with saturated aqueous NH4Cl (1 mL). The
resulting solution was partitioned between EtOAc (3 × 15 mL) and
water (50 mL). The EtOAc phases were combined, dried, and
concentrated to dryness. The residue was purified by column chroma-
tography (silica gel; hexane-acetone, 3:1-1:1) to give the N-acyl
compound 4 as a white solid. This was added to pyridine (1 mL) and
Ac2O (1 mL) and stirred for 8 h at room temperature. Saturated aqueous
NaHCO3 (10 mL) and EtOAc (50 mL) were added. The organic phase
was washed with water (3 × 10 mL) and brine (10 mL), dried, and
concentrated. The residue was purified by column chromatography
(silica gel; hexane-acetone, 4:1-2:1) to give the desired compounds
6a-24a, each as a white solid.
3-Demethoxycarbonyl-3-(propionylamino)methylvindoline (6a). Com-
pound 6a was prepared using propionyl chloride as starting material
1
in 72% yield: H NMR (300 MHz, CDCl3) δ 9.20 (1H, s), 6.88 (1H,
d, J ) 8.1 Hz), 6.32 (1H, dd, J ) 8.1, 2.1 Hz), 6.16 (1H, d, J ) 5.7
Hz), 6.15 (1H, d, J ) 2.1 Hz), 5.89 (1H, dd, J ) 10.2, 4.2 Hz), 5.36
(1H, d, J ) 10.2 Hz), 4.99 (1H, s), 3.79 (3H, s), 3.74 (1H, m), 3.50
(1H, dd, J ) 15.9, 4.5 Hz), 3.39 (1H, m), 3.39 (1H, s), 3.03 (1H, d, J
) 13.2 Hz), 2.86 (3H, s), 2.83 (1H, d, J ) 15.9 Hz), 2.66 (1H, s), 2.53
(1H, dd, J ) 18.0, 9.6 Hz), 2.33-2.19 (2H, m), 2.22 (2H, q, J ) 7.5
Hz), 2.11 (3H, s), 1.36-1.29 (1H, m), 1.15 (3H, t, J ) 7.5 Hz),
1.04-0.98 (1H, m), 0.52 (3H, t, J ) 7.2 Hz); 13C NMR (75 MHz,
CDCl3) δ 173.9, 170.5, 161.0, 154.3, 130.4, 125.6, 124.1, 122.7, 105.2,
96.7, 82.2, 76.9, 75.9, 67.2, 55.4, 52.2, 51.5, 50.8, 44.8, 43.4, 42.8,
40.7, 31.3, 29.8, 21.0, 10.0, 7.5; ESIMS m/z 484.4 [M + 1]+.
3-Demethoxycarbonyl-3-(isobutyrylamino)methylvindoline (7a). Com-
pound 7a was prepared using isobutyryl chloride as starting material
1
in 75% yield: H NMR (300 MHz, CDCl3) δ 9.20 (1H, br s), 6.88
(1H, d, J ) 8.4 Hz), 6.32 (1H, dd, J ) 8.4, 2.1 Hz), 6.19 (1H, d, J )
8.1 Hz), 6.15 (1H, d, J ) 2.1 Hz), 5.89 (1H, dd, J ) 10.2, 3.6 Hz),
5.36 (1H, d, J ) 10.2 Hz), 4.98 (1H, s), 3.79 (3H, s), 3.74 (1H, m),
3.48 (1H, dd, J ) 15.9, 5.4 Hz), 3.38 (1H, m), 3.38 (1H, s), 2.99 (1H,
d, J ) 13.2 Hz), 2.84 (3H, s), 2.83 (1H, d, J ) 15.9 Hz), 2.66 (1H, s),
2.54-2.48 (1H, m), 2.42-2.33 (1H, m), 2.29-2.18 (2H, m), 2.11 (3H,
s), 1.37-1.27 (1H, m), 1.15 (3H, d, J ) 2.1 Hz), 1.13 (3H, d, J ) 2.1
Hz), 1.07-0.98 (1H, m), 0.52 (3H, t, J ) 7.2 Hz); 13C NMR (75 MHz,
CDCl3) δ 171.2, 170.7, 161.2, 154.5, 130.5, 125.8, 124.2, 122.8, 105.4,
96.9, 82.2, 77.0, 76.1, 67.4, 55.5, 52.4, 51.7, 51.0, 45.0, 43.2, 43.0,
40.7, 35.9, 31.4, 21.1, 19.9, 19.7, 7.7; ESIMS m/z 498.3 [M + 1]+.
3-Demethoxycarbonyl-3-(pivaloylamino)methylvindoline (8a). Com-
pound 8a was prepared using pivaloyl chloride as starting material in
a Ditartrate of all compounds used in bioassays.
15.6, 2.0 Hz), 2.59 (1H, s), 2.52-2.47 (1H, m), 2.21-2.15 (2H, m),
1.42-1.37 (1H, m), 0.94-0.88 (1H, m), 0.65 (3H, t, J ) 7.6 Hz); 13
C
NMR (75 MHz, CDCl3) δ 160.8, 154.4, 131.9, 126.5, 122.9, 122.8,
104.2, 96.1, 84.5, 78.4, 75.8, 68.1, 55.4, 52.5, 51.5, 51.3, 49.7, 45.2,
43.6, 41.5, 32.4, 7.9; EIMS m/z 385 [M+], 368, 355, 297, 194, 174,
162, 152, 135, 122, 93.
1
68% yield: H NMR (300 MHz, CDCl3) δ 6.85 (1H, d, J ) 8.4 Hz),
6.44 (1H, d, J ) 8.1 Hz), 6.29 (1H, d, J ) 8.1 Hz), 6.08 (1H, s), 5.84
(1H, dd, J ) 9.9, 4.2 Hz), 5.61 (1H, d, J ) 9.9 Hz), 4.97 (1H, s), 3.74
(3H, s), 3.65 (2H, m), 3.41 (2H, m), 3.33 (1H, s), 3.28 (1H, m), 2.98
(1H, d, J ) 13.2 Hz), 2.84 (3H, s), 2.73 (1H, d, J ) 4.8 Hz), 2.59 (1H,
s), 2.47 (1H, m), 2.16 (2H, m), 2.09 (1H, s), 2.05 (3H, s), 1.92 (3H, s),
3-Demethoxycarbonyl-3-(acetamino)methylvindoline (5a). Com-
pound 4 (385 mg, 1 mmol) was dissolved in pyridine (1 mL) and Ac2O
(1 mL), and the resulting mixture was stirred for 8 h at room
temperature. Saturated aqueous NaHCO3 (10 mL) and EtOAc (50 mL)
were added. The organic phase was washed with water (3 × 10 mL)
and brine (10 mL), dried, and concentrated. The residue was purified
1.30 (1H, m), 1.17 (9H, s), 0.90 (1H, m), 0.57 (3H, t, J ) 7.2 Hz); 13
C
NMR (75 MHz, CDCl3) δ 178.7, 170.3, 161.0, 154.6, 130.8, 126.6,
124.5, 123.0, 105.1, 96.9, 82.4, 76.6, 75.3, 68.5, 55.5, 52.1, 52.0, 51.3,

Cytotoxic AnhydroVinblastine Amide DeriVatiVes
Journal of Natural Products, 2009, Vol. 72, No. 6 1173
Table 2. In Vivo Antitumor Efficacy of 6b, 12b, 24b, 1d, and 1e against Sarcoma 180a
mice (n)
body weight (g)
initial end
route and
schedule
group
dosage (mg/kg)
initial/end
tumor weight (X ( SD)
inhibition rate (%)
NS
6b
16/16
8/8
8/5
8/7
8/5
8/8
8/6
8/8
8/8
21.6
20.6
20.5
20.4
20.5
20.5
20.8
24.1
24.3
31.9
21.4
20.7
22.2
23.9
25.3
23.3
27.5
27.9
1.60 ( 0.27
0.25 ( 0.06
0.16 ( 0.03
0.30 ( 0.05
0.18 ( 0.02
0.67 ( 0.07
0.54 ( 0.17
0.60 ( 0.23
0.49 ( 0.10
10
20
10
20
10
20
10
10
iv d 1, 4
iv d 1
iv d 1, 4
iv d 1
iv d 1, 4
iv d 1
iv, d 1, 4
iv, d 1, 4
84.4b
90.0b
81.3b
88.8b
58.1b
66.3b
62.5b
69.4b
12b
24b
1d
1e
a The in vivo experiment was carried out in Kunming mice bearing sarcoma 180 cells, and the compounds were given intravenously. Ditartrate of all
compounds used in bioassays. b p < 0.01 versus control group.
44.9, 44.0, 43.6, 41.2, 38.9, 32.6, 27.7 (3 C), 20.9, 7.8; ESIMS m/z
512.4 [M + 1]+.
76.0, 67.5, 55.4, 52.3, 51.7, 51.0, 44.8, 44.0, 43.0, 40.7, 31.4, 21.0,
14.8, 7.6, 7.0, 6.9; ESIMS m/z 496.4 [M + 1]+.
3-Demethoxycarbonyl-3-(butyrylamino)methylvindoline (9a). Com-
pound 9a was prepared using butyryl chloride as starting material in
3-Demethoxycarbonyl-3-(cyclobutanecarbonylamino)meth-
ylvindoline (13a). Compound 13a was prepared using cyclobutanecar-
bonyl chloride as starting material in 66% yield: 1H NMR (300 MHz,
CDCl3) δ 9.17 (1H, s), 6.87 (1H, d, J ) 8.1 Hz), 6.32 (1H, dd, J )
8.1, 2.1 Hz), 6.14 (1H, d, J ) 2.1 Hz), 6.07 (1H, d, J ) 7.8 Hz), 5.89
(1H, dd, J ) 10.5, 3.6 Hz), 5.36 (1H, d, J ) 10.5 Hz), 4.98 (1H, s),
3.80-3.73 (1H, m), 3.79 (3H, s), 3.47 (1H, dd, J ) 15.9, 4.8 Hz),
3.42-3.34 (1H, m), 3.37 (1H, s), 3.05-3.00 (2H, m), 2.85 (3H, s),
2.81 (1H, d, J ) 15.9 Hz), 2.66 (1H, s), 2.54-2.48 (1H, m), 2.32-2.12
(6H, m), 2.10 (3H, s), 1.98-1.87 (2H, m), 1.36-1.25 (1H, m),
1.05-0.98 (1H, m),.0.52 (3H, t, J ) 7.2 Hz); 13C NMR (75 MHz,
CDCl3) δ 175.1, 170.6, 161.1, 154.4, 130.4, 125.7, 124.2, 122.8, 105.3,
96.8, 82.3, 77.0, 76.0, 67.3, 55.4, 52.3, 51.6, 50.9, 44.9, 43.3, 42.9,
40.8, 40.1, 31.3, 25.4, 25.4, 21.0, 7.6; ESIMS m/z 510.5 [M + 1]+.
3-Demethoxycarbonyl-3-(benzoylamino)methylvindoline (14a). Com-
pound 14a was prepared using benzoyl chloride as starting material in
1
68% yield: H NMR (300 MHz, CDCl3) δ 9.23 (1H, br s), 6.88 (1H,
d, J ) 8.4 Hz), 6.32 (1H, d, J ) 8.4 Hz), 6.16 (1H, d, J ) 8.1 Hz),
6.15 (1H, s), 5.89 (1H, dd, J ) 10.2, 4.8 Hz), 5.37 (1H, d, J ) 10.2
Hz), 5.01 (1H, s), 3.79 (3H, s), 3.74 (1H, m), 3.48 (1H, dd, J ) 15.9,
5.4 Hz), 3.38 (1H, m), 3.41 (1H, s), 3.04 (1H, d, J ) 14.4 Hz), 2.86
(3H, s), 2.83 (1H, d, J ) 15.9 Hz), 2.67 (1H, s), 2.61-2.52 (1H, m),
2.34 -2.16 (2H, m), 2.19 (2H, t, J ) 7.2 Hz), 2.11 (3H, s), 1.70-1.63
(2H, m), 1.37-1.27 (1H, m), 0.91 (3H, t, J ) 7.2 Hz), 1.03-0.85 (1H,
m), 0.51 (3H, t, J ) 7.2 Hz); 13C NMR (75 MHz, CDCl3) δ 173.2,
170.5, 161.0, 154.3, 130.4, 125.6, 124.1, 122.7, 105.2, 96.7, 82.0, 77.0,
75.9, 67.4, 55.3, 52.2, 51.6, 50.8, 44.8, 43.3, 42.9, 40.6, 38.8, 31.3,
20.9, 19.2, 13.9, 7.5; ESIMS m/z 498.3 [M + 1]+.
3-Demethoxycarbonyl-3-(3′-methylbutyramino)methylvindoline (10a).
Compound 10a was prepared using 3-methylbutyryl chloride as starting
material in 65% yield: 1H NMR (300 MHz, CDCl3) δ 9.25 (1H, br s),
6.87 (1H, d, J ) 8.1 Hz), 6.31 (1H, dd, J ) 8.1, 2.1 Hz), 6.15 (1H, d,
J ) 2.1 Hz), 6.10 (1H, d, J ) 8.1 Hz), 5.89 (1H, dd, J ) 10.5, 3.6
Hz), 5.36 (1H, d, J ) 10.5 Hz), 5.00 (1H, s), 3.79 (3H, s), 3.79-3.72
(1H, m), 3.48 (1H, dd, J ) 15.9, 5.1 Hz), 3.42-3.34 (1H, m), 3.39
(1H, s), 3.03 (1H, d, J ) 13.2 Hz), 2.86 (3H, s), 2.83 (1H, d, J ) 15.9
Hz), 2.63 (1H, s), 2.57-2.48 (1H, m), 2.35-2.16 (3H, m), 2.11 (3H,
s), 2.10 (2H, d, J ) 9.9 Hz), 1.37-1.27 (1H, m), 0.94 (6H, d, J ) 6.3
Hz), 1.03-0.85 (1H, m), 0.51 (3H, t, J ) 7.2 Hz); 13C NMR (75 MHz,
CDCl3) δ 172.7, 170.4, 160.9, 154.2, 130.2, 125.5, 124.0, 122.6, 105.1,
96.5, 81.8, 76.8, 75.8, 67.4, 55.2, 52.1, 51.6, 50.7, 46.1, 44.7, 43.2,
42.8, 40.4, 31.2, 26.0, 22.5, 22.4, 20.9, 7.5; ESIMS m/z 512.4 [M +
1]+.
1
73% yield: H NMR (300 MHz, CDCl3) δ 9.28 (1H, br s), 7.79 (2H,
d, J ) 6.9 Hz), 7.44 (3H, m), 6.91(1H, s), 6.89 (1H, d, J ) 8.1 Hz),
6.33 (1H, d, J ) 8.1 Hz), 6.17 (1H, s), 5.91 (1H, m), 5.40 (1H, d, J )
10.5 Hz), 5.06 (1H, s), 3.93 (1H, m), 3.79 (3H, s), 3.55-3.41 (3H, m),
3.25 (1H, d, J ) 12.9 Hz), 2.91 (3H, s), 2.83 (1H, m), 2.70 (1H, s),
2.55 (1H, m), 2.25 (1H, m), 2.10 (3H, m), 2.09 (3H, s), 1.08-1.01
(1H, m), 0.90-0.84 (1H, m), 0.54 (3H, t, J ) 7.5 Hz); 13C NMR (75
MHz, CDCl3) δ 170.3, 167.6, 161.5, 154.6, 135.4, 132.2, 130.7, 128.6
(2 C), 127.5 (2 C), 125.9, 124.5, 123.1, 105.8, 97.2, 83.1, 77.1, 76.4,
67.5, 55.7, 52.4, 51.5, 51.0, 45.2, 44.2, 43.2, 41.3, 31.6, 21.1, 7.8;
ESIMS m/z 532.3 [M + 1]+.
3-Demethoxycarbonyl-3-(4′-fluorobenzoylamino)methylvindo-
line (15a). Compound 15a was prepared using 4-fluorobenzoyl chloride
as starting material in 73% yield: 1H NMR (300 MHz, CDCl3) δ 9.28
(1H, s), 7.79 (2H, t, J ) 8.1 Hz), 7.11 (2H, t, J ) 8.1 Hz), 6.89 (1H,
d, J ) 8.4 Hz), 6.34 (1H, d, J ) 8.4 Hz), 6.17 (1H, s), 5.91 (1H, dd,
J ) 10.2, 2.4 Hz), 5.40 (1H, d, J ) 10.2 Hz), 5.05 (1H, s), 3.97-3.90
(1H, m), 3.79 (3H, s), 3.53-3.41 (2H, m), 3.45 (1H, s), 3.24 (1H, d,
J ) 13.2 Hz), 2.90 (3H, s), 2.85 (1H, d, J ) 15.9 Hz), 2.70 (1H, s),
2.60-2.51 (1H, m), 2.30-2.17 (2H, m), 2.09 (3H, s), 1.37-1.30 (1H,
m), 1.08-1.01 (1H, m), 0.55 (3H, t, J ) 7.5 Hz); 13C NMR (75 MHz,
CDCl3) δ 170.7, 166.4 (2 C), 161.1, 154.4, 131.0, 130.5, 129.3, 129.2,
125.6, 124.3, 122.8, 115.8, 115.5, 105.5, 97.0, 82.8, 77.0, 76.1, 67.2,
55.5, 52.5, 51.5, 50.9, 44.9, 44.1, 43.0, 41.1, 31.4, 21.0, 7.6; ESIMS
m/z 550.3 [M + 1]+.
3-Demethoxycarbonyl-3-(2′-chlorobenzoylamino)methylvindo-
line (16a). Compound 16a was prepared using 2-chlorobenzoyl chloride
as starting material in 80% yield: 1H NMR (300 MHz, CDCl3) δ 9.16
(1H, s), 7.51 (1H, d, J ) 6.6 Hz), 7.30-7.17 (3H, m), 6.83 (1H, d, J
) 9.0 Hz), 6.80 (1H, d, J ) 9.9 Hz), 6.23 (1H, d, J ) 9.9 Hz, 1 H),
6.07 (1H, s), 5.79 (1H, dd, J ) 10.2, 4.8 Hz), 5.30 (1H, d, J ) 10.2
Hz), 4.98 (1H, s, 1 H), 3.94-3.87 (1H, m), 3.67 (3H, s), 3.42 (1H, s),
3.37 (1H, dd, J ) 15.9, 4.8 Hz), 3.27-3.23 (1H, m), 3.15 (1H, d, J )
13.5 Hz), 2.88 (3H, s), 2.74 (1H, d, J ) 16.5 Hz), 2.57 (1H, s),
2.47-2.38 (1H, m), 2.24-2.07 (2H, m), 2.02 (3H, s), 1.31-1.19 (1H,
m), 1.02-0.89 (1H, m), 0.44 (3H, t, J ) 7.2 Hz); 13C NMR (75 MHz,
CDCl3) δ 170.3, 166.4, 160.9, 154.2, 135.4, 131.0, 130.5, 130.2, 130.0,
129.8, 126.9, 125.5, 124.3, 122.5, 105.1, 96.6, 81.8, 76.8, 75.8, 67.3,
55.1, 52.1, 51.4, 50.7, 44.6, 43.8, 42.7, 40.7, 31.2, 20.8, 7.4; ESIMS
m/z 566.3 [M + 1]+.
3-Demethoxycarbonyl-3-(3′,3′-dimethylbutyramino)methylvindo-
line (11a). Compound 11a was prepared using 3,3-dimethylbutyryl
chloride as starting material in 65% yield: 1H NMR (300 MHz, CDCl3)
δ 9.25 (1H, br s), 6.88 (1H, d, J ) 8.1 Hz), 6.32 (1H, dd, J ) 8.1, 2.1
Hz), 6.15 (1H, d, J ) 2.1 Hz), 6.10 (1H, d, J ) 8.1 Hz), 5.89 (1H, dd,
J ) 9.9, 3.3 Hz), 5.36 (1H, d, J ) 9.9 Hz), 5.02 (1H, s), 3.79 (3H, s),
3.79-3.72 (1H, m), 3.48 (1H, dd, J ) 15.9, 5.1 Hz), 3.42-3.34 (1H,
m), 3.42 (1H, s), 3.03 (1H, d, J ) 13.2 Hz), 2.87 (3H, s), 2.83 (1H, d,
J ) 15.9 Hz), 2.64 (1H, s), 2.57-2.48 (1H, m), 2.35-2.16 (2H, m),
2.12 (3H, s), 2.10 (2H, s), 1.37-1.27 (1H, m), 0.94 (6H, d, J ) 6.3
Hz), 1.03 (9H, s), 0.87-0.80 (1H, m), 0.50 (3H, t, J ) 7.2 Hz); 13C
NMR (75 MHz, CDCl3) δ 171.9, 170.3, 160.9, 154.2, 130.2, 125.6,
124.0, 122.5, 105.0, 96.5, 81.7, 76.9, 75.7, 67.4, 55.2, 52.0, 51.6, 50.7,
50.5, 44.7, 43.1, 42.7, 40.4, 31.2, 30.8, 29.7 (3 C), 20.8, 7.4; ESIMS
m/z 526.3 [M + 1]+.
3-Demethoxycarbonyl-3-(cyclopropanecarbonylamino)meth-
ylvindoline (12a). Compound 12a was prepared using cyclopropan-
1
ecarbonyl chloride as starting material in 62% yield: H NMR (300
MHz, CDCl3) δ 9.02 (1H, s), 6.87 (1H, d, J ) 8.1 Hz), 6.31 (2H, dd,
J ) 8.1, 2.4 Hz), 6.14 (1H, d, J ) 2.4 Hz), 5.89 (1H, d, J ) 10.2, 3.6
Hz), 5.37 (1H, d, J ) 10.2 Hz), 5.01 (1H, s), 3.75 (3H, s), 3.75-3.70
(1H, m), 3.51(1H, dd, J ) 15.9, 4.5 Hz), 3.43 (1H, s), 3.43-3.35 (1H,
m), 3.09 (1H, d, J ) 13.5 Hz), 2.86 (3H, s), 2.82 (1H, d, J ) 15.9 Hz),
2.65 (1H, s), 2.58-2.49 (1H, m), 2.34-2.20 (2H, m), 2.10 (3H, s),
1.44-1.29 (2H, m), 1.18-1.07 (1H, m), 1.00-0.91 (2H, m), 0.76-0.67
(2H, m), 0.52 (3H, t, J ) 7.5 Hz); 13C NMR (75 MHz, CDCl3) δ 173.6,
170.5, 161.1, 154.5, 130.5, 125.7, 124.2, 122.7, 105.2, 96.7, 82.3, 77.2,

1174 Journal of Natural Products, 2009, Vol. 72, No. 6
Shao et al.
3-Demethoxycarbonyl-3-(3′-chlorobenzoylamino)methylvindo-
line (17a). Compound 17a was prepared using 3-chlorobenzoyl chloride
as starting material in 81% yield: 1H NMR (300 MHz, CDCl3) δ 9.29
(1H, br s), 7.77 (1H, s), 7.65 (1H, d, J ) 7.5 Hz), 7.47 (1H, d, J ) 7.5
Hz), 7.37 (1H, t, J ) 7.5 Hz), 6.95 (1H, d, J ) 6.6 Hz), 6.89 (1H, d,
J ) 8.1 Hz), 6.34 (1H, d, J ) 8.1 Hz), 6.18 (1H, s), 5.92 (1H, dd, J )
10.5, 4.5 Hz), 5.40 (1H, d, J ) 10.5 Hz), 5.05 (1H, s), 3.95 (1H, dd,
J ) 13.2, 7.5 Hz), 3.80 (3H, s), 3.54-3.42 (2H, m), 3.44 (1H, s), 3.24
(1H, d, J ) 13.5 Hz), 2.91 (3H, s), 2.84 (1H, d, J ) 16.5 Hz), 2.71
(1H, s), 2.58-2.53 (1H, m), 2.29-2.23 (2H, m), 2.10 (3H, s),
chloride as starting material in 80% yield: 1H NMR (300 MHz, CDCl3)
δ 9.04 (1H, br s), 7.19 (2H, d, J ) 8.4 Hz), 6.87 (2H, d, J ) 8.4 Hz),
6.85 (1H, d, J ) 8.1 Hz), 6.32 (1H, dd, J ) 8.1, 2.1 Hz), 6.16 (1H, d,
J ) 8.1 Hz), 6.02 (1H, s), 5.87 (1H, dd, J ) 10.2, 3.6 Hz), 5.32 (1H,
d, J ) 10.2 Hz), 4.89 (1H, s), 3.86-3.82 (1H, m), 3.81 (3H, s), 3.79
(3H, s), 3.48 (1H, s), 3.48-3.41 (1H, m), 3.35-3.27 (1H, m), 3.16
(1H, s), 2.92 (1H, d, J ) 13.5 Hz), 2.80 (1H, d, J ) 15.9 Hz), 2.64
(3H, s), 2.53-2.44 (1H, m), 2.26-2.00 (2H, m), 2.09 (3H, s),
1.39-1.27 (1H, m), 1.03-0.91 (1H, m), 0.51 (3H, t, J ) 7.2 Hz); 13
C
NMR (75 MHz, CDCl3) δ 171.5, 170.9, 161.0, 158.8, 154.3, 130.7,
130.4, 127.3, 125.8, 124.2, 122.7, 114.4, 105.4, 97.5, 82.9, 76.4, 75.7,
67.1, 55.5, 55.3, 52.3, 51.5, 50.8, 45.0, 43.5, 43.2, 41.1, 31.2, 21.1,
7.6; ESIMS m/z 562.3 [M + 1]+.
1.37-1.30 (1H, m), 1.08-1.01 (1H, m), 0.55 (3H, t, J ) 7.5 Hz); 13
C
NMR (75 MHz, CDCl3) δ 170.7, 166.1, 161.2, 154.4, 136.6, 134.8,
131.5, 130.5, 130.0, 127.4, 125.6, 125.1, 124.3, 122.9, 105.7, 97.0,
82.9, 76.9, 76.1, 67.2, 55.5, 52.5, 51.4, 51.0, 44.9, 44.1, 43.0, 41.2,
31.4, 21.1, 7.6; ESIMS m/z 566.4 [M + 1]+.
3-Demethoxycarbonyl-3-(phenylacetylamino)methylvindoline (23a).
Compound 23a was prepared using phenylacetyl chloride as starting
material in 76% yield: 1H NMR (300 MHz, CDCl3) δ 7.26-7.20 (5H,
m), 6.79 (1H, d, J ) 8.4 Hz), 6.25 (1H, d, J ) 8.4 Hz), 6.10 (1H, d,
J ) 9.0 Hz), 5.95 (1H, s), 5.81 (1H, dd, J ) 10.2, 4.2 Hz), 5.26 (1H,
d, J ) 10.2 Hz), 4.83 (1H, s), 3.79-3.73 (1H, m), 3.73 (3H, s), 3.48
(2H, s), 3.38 (1H, dd, J ) 15.9, 5.1 Hz), 3.30-3.20 (1H, m), 3.08
(1H, s), 2.86 (1H, d, J ) 13.8 Hz), 2.74 (1H, d, J ) 15.9 Hz), 2.55
(3H, s), 2.47-2.38 (1H, m), 2.20-1.93 (2H, m), 2.02 (1H, s),
3-Demethoxycarbonyl-3-(4′-chlorobenzoylamino)methylvindo-
line (18a). Compound 18a was prepared using 4-chlorobenzoyl chloride
as starting material in 78% yield: 1H NMR (300 MHz, CDCl3) δ 9.21
(1H, br s), 7.64 (2H, d, J ) 8.7 Hz), 7.28 (1H, d, J ) 8.7 Hz), 6.89
(1H, d, J ) 7.2 Hz), 6.81 (1H, d, J ) 8.1 Hz), 6.25 (1H, d, J ) 8.1
Hz), 6.08 (1H, s), 5.83 (1H, dd, J ) 9.9, 4.5 Hz), 5.31 (1H, d, J ) 9.9
Hz), 4.97 (1H, s), 3.88-3.80 (1H, m), 3.68 (3H, s), 3.43-3.31 (2H,
m), 3.37 (1H, s), 3.18 (1H, d, J ) 13.5 Hz), 2.81 (3H, s), 2.76 (1H, d,
J ) 16.5 Hz), 2.62 (1H, s), 2.47 (1H, q, J ) 9.3 Hz), 2.24-2.13 (2H,
m), 1.97 (3H, s), 1.28-1.19 (1H, m), 1.02-0.93 (1H, m), 0.46 (3H, t,
J ) 7.2 Hz); 13C NMR (75 MHz, CDCl3) δ 170.5, 166.1, 161.1, 154.2,
137.4, 133.1, 130.3, 128.7 (2 C), 128.3 (2 C), 125.5, 124.1, 122.7,
105.4, 96.9, 82.9, 76.9, 75.9, 67.1, 55.2, 52.4, 51.4, 50.8, 44.7, 44.1,
42.8, 41.0, 31.2, 20.9, 7.5; ESIMS m/z 566.4 [M + 1]+.
1.30-1.19 (1H, m), 0.94-0.89 (1H, m), 0.45 (3H, t, J ) 7.2 Hz); 13
C
NMR (75 MHz, CDCl3) δ 171.0, 170.8, 161.0, 154.3, 135.4, 130.4,
129.6 (2 C), 129.0 (2 C), 127.2, 125.8, 124.2, 122.8, 105.5, 97.4, 82.9,
76.4, 75.8, 67.1, 55.5, 52.3, 51.4, 50.8, 45.0, 44.1, 43.6, 42.9, 41.0,
31.3, 21.1, 7.6; ESIMS m/z 546.4 [M + 1]+.
3-Demethoxycarbonyl-3-(3′,4′-methylenedioxybenzoylamino)me-
thylvindoline (24a). Compound 24a was prepared using 3,4-methylene-
dioxybenzoyl chloride as starting material in 74% yield: 1H NMR (300
MHz, CDCl3) δ 9.26 (1H, br s), 7.32 (1H, d, J ) 8.1 Hz), 7.28 (1H,
s), 6.88 (1H, d, J ) 8.4 Hz), 6.83 (1H, d, J ) 8.1 Hz), 6.82 (1H, s),
6.33 (1H, d, J ) 8.4 Hz), 6.16 (1H, d, J ) 2.1 Hz), 6.02 (2H, s), 5.91
(1H, dd, J ) 10.2, 3.6 Hz), 5.39 (1H, d, J ) 10.2 Hz), 5.04 (1H, s),
3.89 (1H, dd J ) 13.5, 8.1 Hz), 3.79 (3H, s), 3.54-3.42 (2H, m), 3.45
(1H, s), 3.22 (1H, d, J ) 13.5 Hz), 2.89 (3H, s), 2.83 (1H, d, J ) 15.9
Hz), 2.69 (1H, s), 2.60-2.51 (1H, m), 2.33-2.10 (2H, m), 2.09 (3H,
s), 1.37-1.25 (1H, m), 1.07-1.00 (1H, m), 0.54 (3H, t, J ) 7.5 Hz);
13C NMR (75 MHz, CDCl3) δ 170.2, 166.3, 160.7, 153.9, 149.9, 147.6,
130.0, 128.5, 125.3, 123.9, 122.5, 121.1, 107.7, 107.2, 105.0, 101.4,
96.4, 82.3, 76.6, 75.7, 66.8, 55.0, 52.0, 51.1, 50.5, 44.5, 43.7, 42.5,
40.6, 30.9, 20.6, 7.3; ESIMS m/z 576.4 [M + 1]+.
General Procedure for the Preparation of 5b-24b. Catharanthine
tartrate (486 mg, 1 mmol) and anhydrous ferric chloride (486 mg, 3
mmol) were combined in a mixture of glycine buffer (containing 320
mg of glycine and 250 mg of sodium chloride in 40 mL of water) and
hydrochloride acid (40 mL, 0.1 N) under a nitrogen atmosphere. After
10 min of stirring at room temperature, compounds 5a-24a (1 mmol)
were added. After 8 h of stirring at room temperature, sodium
borohydride (80 mg) in ammonium hydroxide (8 mL) was added
dropwise at 0 °C, and it was left to react for 15 min. The reaction
mixture was extracted with CH2Cl2 (4 × 20 mL), and the organic phase
was filtered through Celite and concentrated at reduced pressure. Then
each residue was purified by column chromatography on silica gel (100:
1-80:1, CHCl3-MeOH) to afford each compound (5b-24b) as a white
solid in over 50% yield.
3-Demethoxycarbonyl-3-(pyridine-4′-carbonylamino)methylvindo-
line (19a). Compound 19a was prepared using pyridine-4-carbonyl
chloride as starting material in 82% yield: 1H NMR (300 MHz, CDCl3)
δ 9.37 (1H, br s), 8.74 (2H, d, J ) 3.9 Hz), 7.62 (2H, d, J ) 3.9 Hz),
7.10 (1H, d, J ) 5.4 Hz), 6.90 (1H, d, J ) 8.4 Hz), 6.35 (1H, d, J )
8.4 Hz), 6.18 (1H, s), 5.91 (1H, dd, J ) 10.2, 4.5 Hz), 5.40 (1H, d, J
) 10.2 Hz), 5.05 (1H, s), 3.99-3.92 (1H, m), 3.80 (3H, s), 3.52-3.42
(2H, m), 3.42 (1H, s), 3.26 (1H, d, J ) 13.5 Hz), 2.90 (3H, s), 2.86
(1H, d, J ) 16.5 Hz), 2.72 (1H, s), 2.57 (1H, q, J ) 9.6 Hz), 2.26-2.17
(2H, m), 2.09 (3H, s), 1.31-1.25 (1H, m), 1.08-1.03 (1H, m), 0.56
(3H, t, J ) 7.2 Hz); 13C NMR (75 MHz, CDCl3) δ 170.7, 165.4, 161.2,
154.3, 150.7 (2 C), 141.9, 130.4, 125.5, 124.4, 122.9, 120.9 (2 C),
105.8, 97.2, 83.1, 76.8, 76.0, 67.1, 55.5, 52.6, 51.4, 50.9, 44.9, 44.2,
43.0, 41.4, 31.4, 21.1, 7.6; ESIMS m/z 533.4 [M + 1]+.
3-Demethoxycarbonyl-3-(4′-nitrobenzoylamino)methylvindoline (20a).
Compound 20a was prepared using 4-nitrobenzoyl chloride as starting
1
material in 80% yield: H NMR (300 MHz, CDCl3) δ 9.37 (1H, s),
8.29 (2H, d, J ) 8.7 Hz), 7.93 (2H, d, J ) 8.7 Hz), 7.11 (1H, d, J )
5.4 Hz), 6.90 (1H, d, J ) 7.8 Hz), 6.36 (1H, d, J ) 7.8 Hz), 6.18 (1H,
s), 5.91 (1H, dd, J ) 10.2, 4.5 Hz), 5.40 (1H, d, J ) 10.2 Hz), 5.05
(1H, s), 4.01-3.94 (1H, m), 3.80 (3H, s), 3.53-3.42 (2H, m), 3.43
(1H, s), 3.27 (1H, d, J ) 13.5 Hz), 2.91 (3H, s), 2.86 (1H, d, J ) 16.5
Hz), 2.73 (1H, s), 2.62-2.53 (1H, m), 2.33-2.17 (2H, m), 2.09 (3H,
s), 1.36-1.25 (1H, m), 1.08-1.01 (1H, m), 0.56 (3H, t, J ) 7.2 Hz);
13C NMR (75 MHz, CDCl3) δ 170.3, 164.9, 160.9, 153.9, 149.2, 140.1,
130.0, 127.9 (2 C), 125.3, 124.0, 123.4 (2 C), 120.6, 105.3, 96.8, 82.8,
76.5, 75.6, 66.8, 55.0, 52.3, 51.0, 50.5, 44.5, 44.1, 42.6, 41.0, 31.0,
20.6, 7.2; ESIMS m/z 577.3 [M + 1]+.
3-Demethoxycarbonyl-3-(acetamino)methylanhydrovinblastine (5b):
1
[R]20 +63.4 (c 0.30, CHCl3); H NMR (300 MHz, CDCl3) δ 9.33
3-Demethoxycarbonyl-3-(2′-methoxybenzoylamino)methylvin-
doline (21a). Compound 21a was prepared using 2-methoxybenzoyl
chloride as starting material in 82% yield: 1H NMR (300 MHz, CDCl3)
δ 9.09 (1H, s), 8.45 (1H, d, J ) 7.8 Hz), 8.15 (1H, d, J ) 7.5 Hz),
7.43 (1H, t, J ) 7.8 Hz), 7.06 (1H, t, J ) 7.8 Hz), 6.97 (1H, d, J ) 7.8
Hz), 6.88 (1H, d, J ) 8.4 Hz), 6.30 (1H, d, J ) 8.4 Hz), 6.14 (1H, s),
5.88 (1H, dd, J ) 10.2, 4.8 Hz), 5.37 (1H, d, J ) 10.2 Hz), 5.07 (1H,
s), 4.01-3.93 (1H, m), 3.94 (3H, s), 3.79 (3H, s), 3.54-3.30 (2H, m),
3.49 (1H, s), 3.26 (1H, d, J ) 13.5 Hz), 2.88 (3H, s), 2.83 (1H, d, J )
16.5 Hz), 2.65 (1H, s), 2.58-2.49 (1H, m), 2.35-2.16 (2H, m), 2.01
(3H, s), 1.42-1.35 (1H, m), 1.08-1.01 (1H, m), 0.52 (3H, t, J ) 7.2
Hz); 13C NMR (75 MHz, CDCl3) δ 170.1, 164.8, 160.6, 157.3, 154.1,
132.2, 131.4, 130.0, 125.5, 123.7, 122.3, 121.6, 120.6, 111.2, 104.5,
96.2, 81.8, 76.8, 75.6, 67.2, 55.7, 54.8, 51.8, 51.3, 50.4, 44.5, 43.7,
42.5, 40.1, 31.1, 20.4, 7.2; ESIMS m/z 562.3 [M + 1]+.
D
(1H, s), 7.95 (1H, s), 7.45 (1H, d, J ) 7.2 Hz), 7.06 (3H, m), 6.52
(1H, s), 6.13 (1H, s), 6.10 (1H, d, J ) 8.7 Hz), 5.81 (1H, dd, J ) 10.2,
4.5 Hz), 5.42 (1H, d, J ) 4.8 Hz), 5.35 (1H, d, J ) 10.2 Hz), 4.96
(1H, s), 3.76 (3H, s), 3.55 (3H, s), 2.82 (3H, s), 2.55 (1H, s), 2.07 (3H,
s), 1.93 (3H, s), 1.43 (1H, m), 0.93 (3H, t, J ) 7.2 Hz), 0.73 (3H, t, J
) 6.9 Hz); 13C NMR (75 MHz, CDCl3) δ 174.9, 170.9, 170.5, 158.1,
153.7, 139.9, 135.2, 131.1, 130.1, 129.6, 124.7, 124.0, 123.7, 123.7,
122.5, 121.8, 119.1, 118.5, 117.4, 110.7, 95.3, 82.4, 76.8, 76.0, 66.2,
56.0, 55.6, 54.7, 52.8, 52.6, 52.2, 50.3 (2 C), 46.0, 45.3, 43.5, 42.9,
40.9, 34.5, 32.9, 31.6, 28.0, 25.6, 23.5, 21.1, 12.4, 8.4; ESIMS m/z
806.5 [M + 1]+; HRESIMS m/z C47H60N5O7 [M + H]+ calcd for
806.4493, found 806.4495.
3-Demethoxycarbonyl-3-(propionylamino)methylanhy-
1
drovinblastine (6b): [R]20 +65.1 (c 0.28, CHCl3); H NMR (300
D
3-Demethoxycarbonyl-3-(4′-methoxybenzoylamino)methylvin-
doline (22a). Compound 22a was prepared using 4-methoxybenzoyl
MHz, CDCl3) δ 9.40 (1H, s), 8.00 (1H, s), 7.45 (1H, d, J ) 7.5 Hz),
7.13 (3H, m), 6.53 (1H, s), 6.18 (1H, d, J ) 7.8 Hz), 6.14 (1H, s),

Cytotoxic AnhydroVinblastine Amide DeriVatiVes
Journal of Natural Products, 2009, Vol. 72, No. 6 1175
5.81 (1H, dd, J ) 10.2, 3.9 Hz), 5.43 (1H, d, J ) 6.3 Hz), 5.35 (1H,
d, J ) 10.2 Hz), 4.95 (1H, s), 3.76 (3H, s), 3.55 (3H, s), 3.28 (1H, s),
2.81 (3H, s), 2.56 (1H, s), 2.16 (2H, q, J ) 7.8 Hz), 2.08 (3H, s), 1.87
(2H, q, J ) 7.8 Hz), 1.41 (1H, m), 1.08 (3H, t, J ) 7.8 Hz), 0.93 (3H,
t, J ) 7.8 Hz), 0.74 (3H, t, J ) 6.9 Hz); 13C NMR (75 MHz, CDCl3)
δ 174.9, 174.3, 170.9, 158.0, 153.6, 139.8, 135.0, 130.9, 130.0, 129.5,
124.6, 123.8, 123.6, 123.6, 122.4, 121.7, 120.0, 118.4, 117.4, 110.7,
95.2, 82.2, 77.0, 76.0, 65.9, 56.0, 55.5, 54.6, 52.7, 52.5, 51.9, 50.1 (2
C), 45.8, 45.3, 43.1, 42.8, 40.7, 34.3, 32.8, 31.5, 29.9, 28.0, 25.3, 21.0,
12.3, 10.1, 8.4; ESIMS m/z 820.4 [M + 1]+; HRESIMS m/z C48H62N5O7
[M + H]+ calcd for 820.4649, found 806.4650.
3-Demethoxycarbonyl-3-(3′,3′-dimethylbutyramino)methylanhy-
1
drovinblastine (11b): [R]20 +68.1 (c 0.30, CHCl3); H NMR (300
D
MHz, CDCl3) δ 9.42 (1H, s), 8.01 (1H, s), 7.52 (1H, d, J ) 7.8 Hz),
7.17-7.09 (3H, m), 6.58 (1H, s), 6.19 (1H, s), 6.07 (1H, d, J ) 7.8
Hz), 5.88 (1H, dd, J ) 10.2, 3.6 Hz), 5.49 (1H, d, J ) 6.0 Hz), 5.41
(1H, d, J ) 10.2 Hz), 5.02 (1H, s), 3.83 (3H, s), 3.79-3.69 (1H, m),
3.62 (3H, s), 3.32 (1H, s), 2.89 (3H, s), 2.81 (1H, d, J ) 14.1 Hz),
2.59 (1H, s), 2.15 (3H, s), 2.10 (2H, s), 1.94 (2H, q, J ) 7.2 Hz),
1.53-1.46 (1H, m), 1.49 (1H, m), 1.30-1.19 (1H, m), 1.02 (9H, s),
0.99 (3H, t, J ) 7.5 Hz), 0.80 (3H, t, J ) 7.2 Hz); 13C NMR (75 MHz,
CDCl3) δ 174.7, 172.1, 170.6, 158.0, 153.5, 140.1, 135.0, 131.0, 130.0,
129.5, 124.5, 123.9, 123.7, 123.6, 122.3, 121.7, 118.9, 118.4, 117.4,
110.5, 95.1, 81.9, 77.0, 75.9, 66.2, 55.9, 55.5, 54.6, 52.5, 52.4, 52.3,
50.6, 50.4, 50.1, 45.9, 45.3, 42.9, 42.8, 40.5, 34.4, 33.0, 31.4, 31.0,
29.9 (3 C), 29.7, 25.8, 21.0, 12.3, 8.3; ESIMS m/z 862.4 [M + 1]+;
HRESIMS m/z C51H68N5O7 [M + H]+ calcd for 862.5119, found
862.5114.
3-Demethoxycarbonyl-3-(isobutyrylamino)methylanhydrovinblastine
1
(7b): [R]20 +64.0 (c 0.30, CHCl3); H NMR (300 MHz, CDCl3) δ
D
9.43 (1H, s), 8.01 (1H, s), 7.52 (1H, d, J ) 7.5 Hz), 7.16 (3H, m),
6.61 (1H, s), 6.21 (1H, s), 6.19 (1H, s), 5.88 (1H, dd, J ) 10.2, 3.6
Hz), 5.47 (1H, d, J ) 6.0 Hz), 5.42 (1H, d, J ) 10.2 Hz), 5.02 (1H, s),
3.82 (3H, s), 3.62 (3H, s), 3.34 (1H, s), 2.86 (3H, s), 2.62 (1H, s), 2.15
(3H, s), 1.96 (2H, q, J ) 7.5 Hz), 1.49 (1H, m), 1.15 (3H, d, J ) 1.8
Hz), 1.14 (3H, d, J ) 1.8 Hz), 0.99 (3H, t, J ) 7.5 Hz), 0.80 (3H, t,
J ) 7.2 Hz); 13C NMR (75 MHz, CDCl3) δ 177.3, 174.3, 170.9, 158.0,
153.6, 139.8, 135.1, 131.0, 130.0, 129.5, 124.7, 123.9, 123.7, 123.7,
122.5, 121.7, 119.1, 118.5, 117.2, 110.7, 95.2, 82.1, 77.0, 76.0, 66.0,
56.0, 55.5, 54.6, 52.7, 52.6, 51.9, 50.8, 50.2, 45.8, 45.4, 42.8, 42.8,
40.6, 35.8, 34.4, 32.7, 31.5, 28.0, 25.3, 21.0, 19.8, 19.7, 12.3, 8.4;
ESIMS m/z 834.4 [M + 1]+; HRESIMS m/z C49H64N5O7 [M + H]+
calcd for 834.4806, found 834.4805.
3-Demethoxycarbonyl-3-(cyclopropanecarbonylamino)meth-
1
ylanhydrovinblastine (12b): [R]20 +65.0 (c 0.20, CHCl3); H NMR
D
(300 MHz, CDCl3) δ 9.38 (1H, s), 8.01 (1H, s), 7.52 (1H, d, J ) 7.5
Hz), 7.19-7.09 (3H, m), 6.61 (1H, s), 6.31 (1H, d, J ) 7.2 Hz), 6.19
(1H, s), 5.88 (1H, dd, J ) 10.2, 3.6 Hz), 5.48 (1H, d, J ) 6.0 Hz),
5.42 (1H, d, J ) 10.2 Hz), 5.04 (1H, s), 3.83 (3H, s), 3.77-3.70 (1H,
m), 3.62 (3H, s), 3.54 (1H, d, J ) 16.5 Hz), 3.40 (1H, s), 2.89 (3H, s),
2.62 (1H, s), 2.15 (3H, s), 1.93 (2H, q, J ) 7.5 Hz), 1.49 (1H, m),
1.00 (3H, t, J ) 7.5 Hz), 0.93 (2H, m), 0.81 (3H, t, J ) 7.2 Hz), 0.71
(2H, m); 13C NMR (75 MHz, CDCl3) δ 174.9, 173.6, 170.7, 158.0,
153.6, 140.1, 135.1, 131.1, 130.1, 129.5, 124.5, 123.9, 123.6, 123.6,
122.3, 121.8, 118.9, 118.4, 117.5, 110.5, 95.1, 82.4, 77.2, 76.0, 66.1,
56.0, 55.5, 54.6, 52.7, 52.4, 52.4, 50.2 (2 C), 46.0, 45.2, 43.7, 42.8,
40.7, 34.4, 33.1, 31.5, 27.9, 26.0, 21.0, 14.8, 12.4, 8.4, 7.0, 6.9; ESIMS
m/z 832.3 [M + 1]+; HRESIMS m/z C49H62N5O7 [M + H]+ calcd for
832.4649, found 832.4651.
3-Demethoxycarbonyl-3-(pivaloylamino)methylanhydrovin-
blastine (8b): [R]20 +64.1 (c 0.34, CHCl3); 1H NMR (300 MHz,
D
DMSO-d6) δ 9.93 (1H, s), 8.56 (1H, s), 7.44 (1H, d, J ) 8.1 Hz), 7.29
(1H, d, J ) 8.1 Hz), 6.98 (2H, m), 6.62 (2H, s), 6.55 (1H, s), 5.79
(1H, m), 5.50 (1H, d, J ) 5.7 Hz), 5.43 (1H, d, J ) 10.2 Hz), 4.78
(1H, s), 3.78 (3H, s), 3.58 (3H, s), 2.86 (3H, s), 2.08 (3H, s), 1.43 (1H,
m), 1.10 (9H, s), 0.96 (3H, t, J ) 7.2 Hz), 0.65 (3H, t, J ) 6.9 Hz);
13C NMR (75 MHz, DMSO-d6) δ 176.8, 174.2, 170.5, 157.4, 153.2,
139.9, 135.5, 131.3, 130.4, 128.6, 124.3, 123.8, 123.6, 121.9, 121.1,
118.1, 117.6, 115.2, 111.7, 95.5, 81.9, 76.1, 75.5, 64.1, 56.2, 55.0, 54.2,
52.6, 52.5, 52.0, 49.4, 48.7, 46.0, 45.4, 42.5, 42.0, 40.5, 40.3, 38.2,
35.2, 32.9, 31.4, 27.2 (3 C), 27.1, 26.6, 20.8, 12.2, 7.9; ESIMS m/z
848.4 [M + 1]+; HRESIMS m/z C50H66N5O7 [M + H]+ calcd for
848.4962, found 848.4965.
3-Demethoxycarbonyl-3-(cyclobutanecarbonylamino)methy-
lanhydrovinblastine (13b): [R]20D +68.0 (c 0.35, CHCl3); 1H NMR (300
MHz, CDCl3) δ 9.38 (1H, s), 8.01 (1H, s), 7.52 (1H, d, J ) 7.8 Hz),
7.16-7.08 (3H, m), 6.60 (1H, s), 6.19 (1H, s), 6.08 (1H, d, J ) 7.8
Hz), 5.87 (1H, dd, J ) 10.2, 3.6 Hz), 5.47 (1H, d, J ) 6.0 Hz), 5.41
(1H, d, J ) 10.2 Hz), 5.01 (1H, s), 3.82 (3H, s), 3.62 (3H, s), 3.31
(1H, s), 2.87 (3H, s), 2.62 (1H, s), 2.14 (3H, s), 1.98-1.87 (2H, m),
1.48 (1H, m), 0.99 (3H, t, J ) 7.5 Hz), 0.80 (3H, t, J ) 7.5 Hz); 13C
NMR (75 MHz, CDCl3) δ 175.1, 174.7, 170.7, 158.0, 153.5, 140.0,
135.0, 131.0, 130.0, 129.5, 124.5, 123.9, 123.6, 123.6, 122.3, 121.7,
118.9, 118.4, 117.3, 110.5, 95.1, 82.2, 76.9, 75.9, 66.0, 55.9, 55.5, 54.5,
52.6, 52.4, 52.2, 50.1 (2 C), 45.9, 45.2, 43.0, 42.7, 40.6, 40.0, 34.3,
32.9, 31.4, 27.8, 25.7, 25.3 (2 C), 21.0, 18.2, 12.3, 8.3; ESIMS m/z
846.4 [M + 1]+; HRESIMS m/z C50H64N5O7 [M + H]+ calcd for
846.4806, found 846.4809.
3-Demethoxycarbonyl-3-(butyrylamino)methylanhydrovinb-
lastine (9b): [R]20 +63.6 (c 0.30, CHCl3); 1H NMR (300 MHz,
D
CDCl3) δ 9.41 (1H, s), 8.02 (1H, s), 7.52 (1H, d, J ) 7.8 Hz), 7.19-7.09
(3H, m), 6.59 (1H, s), 6.19 (1H, s), 6.15 (1H, d, J ) 7.8 Hz), 5.89
(1H, dd, J ) 10.2, 6.0 Hz), 5.49 (1H, d, J ) 5.7 Hz), 5.41 (1H, d, J )
10.2 Hz), 5.02 (1H, s), 3.83 (3H, s), 3.79-3.70 (1H, m), 3.62 (3H, s),
3.55 (1H, d, J ) 16.5 Hz), 3.35 (1H, s), 2.88 (3H, s), 2.62 (1H, s),
2.18 (2H, t, J ) 7.5 Hz), 2.15 (3H, s), 1.94 (2H, q, J ) 7.2 Hz),
1.70-1.60 (2H, m), 1.53-1.45 (1H, m), 1.30-1.20 (1H, m), 1.00 (3H,
3-Demethoxycarbonyl-3-(benzoylamino)methylanhydrovinb-
t, J ) 7.5 Hz), 0.94 (3H, t, J ) 7.5 Hz), 0.80 (3H, t, J ) 7.2 Hz); 13
C
lastine (14b): [R]20 +60.0 (c 0.30, CHCl3); H NMR (300 MHz,
1
D
NMR (75 MHz, CDCl3) δ 174.9, 173.4, 170.8, 158.0, 153.6, 140.0,
135.1, 131.0, 130.0, 129.6, 124.7, 123.9, 123.7, 123.7, 122.4, 121.7,
119.0, 118.5, 117.5, 110.6, 95.2, 82.1, 77.0, 76.0, 66.0, 56.0, 55.6, 54.7,
52.7, 52.5, 52.2, 50.2 (2 C), 45.9, 45.4, 43.0, 42.8, 40.7, 39.0, 34.4,
33.0, 31.5, 27.9, 25.7, 21.2, 19.4, 14.0, 12.4, 8.4; ESIMS m/z 834.4
[M + 1]+; HRESIMS m/z C49H64N5O7 [M + H]+ calcd for 834.4806,
found 834.4807.
DMSO-d6) δ 9.90 (1H, s), 8.64 (1H, s), 7.78 (2H, d, J ) 7.8 Hz), 7.49
(3H, m), 7.29 (1H, d, J ) 7.8 Hz), 6.98 (3H, m), 6.62 (1H, s), 6.56
(1H, s), 5.75 (1H, dd, J ) 10.2, 4.5 Hz), 5.47 (2H, m), 4.86 (1H, s),
3.80 (3H, s), 3.58 (3H, s), 2.90 (3H, s), 2.09 (3H, s), 0.95 (3H, t, J )
7.2 Hz), 0.66 (3H, t, J ) 6.9 Hz); 13C NMR (75 MHz, DMSO-d6) δ
173.9, 173.6, 170.6, 166.38, 157.3, 153.6, 137.6, 137.0, 135.6, 134.6,
131.3 (2 C), 130.6, 128.5 (2 C), 127.0 (2 C), 124.4, 123.8 (2 C), 121.4,
121.0, 118.4, 117.7, 113.2, 112.7, 111.9, 95.3, 81.2, 76.6, 75.8, 63.8,
56.3, 55.0, 53.8, 52.6, 52.2, 49.5 (2 C), 48.7, 45.2 (2 C), 43.1, 42.1,
38.7, 34.8, 31.4, 31.4, 30.7, 27.2, 20.8, 11.8, 7.9; ESIMS m/z 868.3
[M + 1]+; HRESIMS m/z C52H62N5O7 [M + H]+ calcd for 868.4649,
found 868.4645.
3-Demethoxycarbonyl-3-(3′-methylbutyramino)methylanhy-
1
drovinblastine (10b): [R]20 +66.0 (c 0.27, CHCl3); H NMR (300
D
MHz, CDCl3) δ 9.40 (1H, s), 8.03 (1H, s), 7.53 (1H, d, J ) 7.8 Hz),
7.19-7.09 (3H, m), 6.58 (1H, s), 6.19 (1H, s), 6.15 (1H, d, J ) 7.8
Hz), 5.89 (1H, dd, J ) 10.2, 6.0 Hz), 5.49 (1H, d, J ) 5.7 Hz), 5.41
(1H, d, J ) 10.2 Hz), 5.02 (1H, s), 3.83 (3H, s), 3.79-3.72 (1H, m),
3.62 (3H, s), 3.55 (1H, d, J ) 16.5 Hz), 3.35 (1H, s), 2.88 (3H, s),
2.62 (1H, s), 2.15 (3H, s), 2.10 (2H, d, J ) 9.9 Hz), 1.95 (2H, q, J )
7.2 Hz), 1.69-1.59 (2H, m), 1.53-1.45 (1H, m), 1.31-1.21 (1H, m),
0.99 (3H, t, J ) 7.5 Hz), 0.94 (6H, d, J ) 6.3 Hz), 0.80 (3H, t, J ) 7.2
Hz); 13C NMR (75 MHz, CDCl3) δ 174.8, 172.8, 170.7, 158.0, 153.5,
140.1, 135.1, 131.1, 130.0, 129.5, 124.5, 123.9, 123.6, 123.6, 122.3,
121.8, 118.9, 118.4, 117.5, 110.5, 95.1, 82.0, 77.0, 75.9, 66.2, 55.9,
55.5, 54.6, 52.6, 52.4, 52.4, 50.3, 50.2, 46.2, 46.0, 45.3, 43.0, 42.8,
40.6, 34.4, 33.0, 31.5, 27.9, 26.2, 26.0, 22.6, 22.5, 21.0, 12.4, 8.3;
ESIMS m/z 848.4 [M + 1]+; HRESIMS m/z C50H66N5O7 [M + H]+
calcd for 848.4962, found 848.4960.
3-Demethoxycarbonyl-3-(4′-fluorobenzoylamino)methylanhy-
1
drovinblastine (15b): [R]20 +66.4 (c 0.20, CHCl3); H NMR (300
D
MHz, CDCl3) δ 9.51 (1H, s), 8.03 (1H, s), 7.80 (2H, dd, J ) 8.4, 5.4
Hz), 7.53 (1H, d, J ) 7.5 Hz), 7.19-7.08 (5H, m), 6.90 (1H, d, J )
7.5 Hz), 6.63 (1H, s), 6.21 (1H, s), 5.90 (1H, dd, J ) 10.2, 3.9 Hz),
5.46 (1H, s), 5.44 (1H, d, J ) 10.2 Hz), 5.09 (1H, s), 3.95-3.88 (1H,
m), 3.83 (3H, s), 3.63 (3H, s), 3.53 (1H, d, J ) 16.8 Hz), 3.41 (1H, s),
2.91 (3H, s), 2.85 (1H, d, J ) 16.2 Hz), 2.66 (1H, s), 2.14 (3H, s),
1.93 (2H, q, J ) 7.5 Hz), 1.54-1.49 (1H, m), 0.99 (3H, t, J ) 7.5
Hz), 0.82 (3H, t, J ) 7.2 Hz); 13C NMR (75 MHz, CDCl3) δ 174.9,
171.0, 166.6, 166.5 (d), 158.0, 153.6, 139.9, 135.1, 130.9 (2 C), 130.0,
129.5, 129.3 (d, 2 C), 124.7, 123.9, 123.7, 123.6, 122.5, 121.9, 119.0,

1176 Journal of Natural Products, 2009, Vol. 72, No. 6
Shao et al.
118.5, 117.5, 115.7 (d, 2 C), 110.7, 95.3, 82.7, 77.0, 76.1, 65.9, 56.0,
55.6, 54.7, 52.9, 52.6, 52.0, 50.2, 50.1, 45.8, 45.4, 43.8, 42.8, 41.0,
34.4, 32.8, 31.6, 28.0, 25.5, 21.1, 12.3, 8.4; ESIMS m/z 886.4 [M +
1]+; HRESIMS m/z C52H61FN5O7 [M + H]+ calcd for 886.4555, found
886.4551.
7.94 (2H, d, J ) 8.7 Hz), 7.53 (1H, d, J ) 7.5 Hz), 7.13 (4H, m), 6.64
(1H, s), 6.23 (1H, s), 5.91 (1H, dd, J ) 10.2, 4.2 Hz), 5.47 (1H, s),
5.45 (1H, d, J ) 10.2 Hz), 5.09 (1H, s), 3.91 (1H, m), 3.84 (3H, s),
3.63 (3H, s), 3.43 (1H, s), 2.93 (3H, s), 2.84 (2H, d, J ) 16.5 Hz),
2.68 (1H, s), 2.13 (3H, s), 1.93 (2H, q, J ) 7.2 Hz), 1.51 (1H, m),
1.28 (1H, m), 0.99 (3H, t, J ) 7.5 Hz), 0.82 (3H, t, J ) 7.2 Hz); 13C
NMR (75 MHz, CDCl3)δ 174.8, 171.0, 165.4, 158.1, 153.4, 149.6,
140.3, 140.0, 135.1, 130.9, 130.0, 129.6, 128.2 (2 C), 124.8, 124.0 (2
C), 123.8 (2 C), 123.5, 122.5, 122.3, 119.1, 118.5, 117.6, 110.7, 95.4,
83.0, 76.8, 76.0, 65.8, 56.0, 55.6, 54.5, 53.0, 52.6, 52.2, 50.2, 50.1,
45.9, 45.4, 44.0, 42.9, 41.4, 34.4, 33.0, 31.6, 28.0, 25.7, 21.2, 12.4,
8.4; ESIMS m/z 913.4 [M + 1]+; HRESIMS m/z C52H61N6O9 [M +
H]+ calcd for 913.4500, found 913.4503.
3-Demethoxycarbonyl-3-(2′-chlorobenzoylamino)methylanhy-
1
drovinblastine (16b): [R]20 +65.1 (c 0.30, CHCl3); H NMR (300
D
MHz, CDCl3) δ 9.44 (1H, s), 8.04 (1H, s), 7.60 (1H, dd, J ) 6.6, 2.1
Hz), 7.52 (1H, d, J ) 7.8 Hz), 7.32 (3H, m), 7.13 (3H, m), 6.90 (1H,
d, J ) 7.5 Hz), 6.61 (1H, s), 6.23 (1H, s), 5.88 (1H, dd, J ) 10.2, 4.2
Hz), 5.47 (1H, s), 5.44 (1H, d, J ) 10.2 Hz), 5.09 (1H, s), 3.98 (1H,
m), 3.83 (3H, s), 3.62 (3H, s), 3.46 (1H, s), 3.00 (3H, s), 2.81 (2H, d,
J ) 15.6 Hz), 2.62 (1H, s), 2.18 (3H, s), 1.93 (2H, q, J ) 7.5 Hz),
1.51 (1H, m), 1.26 (1H, m), 0.99 (3H, t, J ) 7.5 Hz), 0.82 (3H, t, J )
6.9 Hz); 13C NMR (75 MHz, CDCl3) δ 174.9, 170.9, 166.9, 158.1,
153.7, 140.0, 135.6, 135.1, 131.3, 130.8, 130.3, 130.0 (2 C), 130.0,
129.6, 127.2, 124.6, 124.0, 123.7, 123.7, 122.4, 121.8, 119.0, 118.5,
117.5, 110.6, 94.7, 82.1, 77.0, 76.1, 66.2, 56.0, 55.6, 54.7, 52.7, 52.5,
52.2, 50.3, 50.2, 46.0, 45.4, 43.8, 42.9, 41.0, 34.5, 33.0, 31.6, 28.0,
25.8, 21.2, 12.4, 8.4; ESIMS m/z 902.5 [M + 1]+; HRESIMS m/z
C52H61ClN5O7 [M + H]+ calcd for 902.4260, found 902.4263.
3-Demethoxycarbonyl-3-(2′-methoxybenzoylamino)methylanhy-
1
drovinblastine (21b): [R]20 +62.1 (c 0.20, CHCl3); H NMR (300
D
MHz, CDCl3) δ 9.27 (1H, s), 8.47 (1H, d, J ) 7.2 Hz), 8.14 (1H, d, J
) 7.5 Hz), 8.05 (1H, s), 7.52 (1H, d, J ) 7.5 Hz), 7.42 (1H, t, J ) 7.5
Hz), 7.14 (3H, m), 7.08 (1H, t, J ) 7.5 Hz), 6.96 (1H, d, J ) 7.5 Hz),
6.61 (1H, s), 6.20 (1H, s), 5.88 (1H, dd, J ) 10.2, 4.2 Hz), 5.49 (1H,
s), 5.45 (1H, d, J ) 10.2 Hz), 5.09 (1H, s), 3.93 (1H, m), 3.90 (3H, s),
3.82 (3H, s), 3.61 (3H, s), 3.44 (1H, s), 2.91 (3H, s), 2.83 (2H, d, J )
15.6 Hz), 2.63 (1H, s), 2.13 (3H, s), 1.93 (2H, q, J ) 7.5 Hz), 1.51
(1H, m), 1.26 (1H, m), 0.99 (3H, t, J ) 7.5 Hz), 0.82 (3H, t, J ) 6.9
Hz); 13C NMR (75 MHz, CDCl3) δ 174.9, 170.9, 165.5, 158.1, 157.7,
153.8, 140.0, 135.1, 132.6, 132.0, 131.3, 130.2, 129.6, 124.6, 124.0,
123.8, 123.7, 122.4, 122.3, 121.5, 121.2, 119.0, 118.5, 117.5, 111.6,
110.6, 95.1, 82.2, 77.3, 76.1, 66.3, 56.1, 56.0, 55.6, 54.7, 52.7, 52.5,
52.3, 50.4, 50.3, 46.0, 45.4, 43.8, 42.9, 40.6, 34.5, 33.0, 31.6, 28.0,
25.8, 21.1, 12.4, 8.4; ESIMS m/z 898.5 [M + 1]+; HRESIMS m/z
C53H64N5O8 [M + H]+ calcd for 898.4755, found 898.4751.
3-Demethoxycarbonyl-3-(3′-chlorobenzoylamino)methylanhy-
1
drovinblastine (17b): [R]20 +65.8 (c 0.32, CHCl3); H NMR (300
D
MHz, CDCl3) δ 9.48 (1H, s), 8.03 (1H, s), 7.77 (1H, s), 7.66 (1H, d,
J ) 7.8 Hz), 7.53 (1H, d, J ) 7.8 Hz), 7.47 (1H, d, J ) 7.8 Hz), 7.36
(1H, t, J ) 7.8 Hz), 7.19-7.09 (3H, m), 6.95 (1H, d, J ) 7.8 Hz),
6.63 (1H, s), 6.22 (1H, s), 5.90 (1H, dd, J ) 9.6, 4.5 Hz), 5.47-5.43
(2H, m), 5.08 (1H, s), 3.93 (1H, dd, J ) 13.5, 7.5 Hz), 3.83 (3H, s),
3.63 (3H, s), 3.53 (1H, d, J ) 16.5 Hz), 3.40 (1H, s), 2.92 (3H, s),
2.85 (1H, d, J ) 15.9 Hz), 2.66 (1H, s), 2.14 (3H, s), 1.93 (2H, q, J )
7.2 Hz), 1.54-1.47 (1H, m), 1.31-1.25 (1H, m), 0.99 (3H, t, J ) 7.5
Hz), 0.82 (3H, t, J ) 7.2 Hz); 13C NMR (75 MHz, CDCl3) δ 174.8,
170.9, 166.1, 158.1, 153.5, 140.0, 136.6, 135.1, 134.7, 131.5, 131.0,
130.0 (2 C), 129.5, 127.3, 125.1, 124.7, 123.9, 122.7, 123.6, 122.4,
122.1, 119.0, 118.5, 117.5, 110.6, 95.4, 82.8, 76.8, 76.0, 65.9, 56.0,
55.6, 54.6, 52.9, 52.5, 52.2, 50.1 (2 C), 45.9, 45.3, 43.8, 42.8, 41.1,
34.4, 33.0, 31.5, 27.9, 25.8, 21.1, 12.4, 8.4; ESIMS m/z 902.5 [M +
1]+; HRESIMS m/z C52H61ClN5O7 [M + H]+ calcd for 902.4260, found
902.4262.
3-Demethoxycarbonyl-3-(4′-methoxybenzoylamino)methylanhy-
1
drovinblastine (22b): [R]20 +63.1 (c 0.17, CHCl3); H NMR (300
D
MHz, CDCl3) δ 9.24 (1H, s), 8.00 (1H, s), 7.52 (1H, d, J ) 7.8 Hz),
7.20 (2H, d, J ) 8.7 Hz), 7.14-7.08 (3H, m), 6.87 (2H, d, J ) 8.7
Hz), 6.58 (1H, s), 6.17 (1H, d, J ) 8.4 Hz), 6.10 (1H, s), 5.85 (1H, dd,
J ) 10.2, 4.2 Hz), 5.46 (1H, s), 5.38 (1H, d, J ) 10.2 Hz), 4.95 (1H,
s), 3.84 (3H, s), 3.79 (3H, s), 3.63 (1H, s), 3.12 (1H, s), 2.91 (1H, d,
J ) 13.5 Hz), 2.80 (1H, d, J ) 15.9 Hz), 2.68 (3H, s), 2.59 (1H, s),
2.13 (3H, s), 1.93 (2H, q, J ) 7.2 Hz), 1.45 (1H, m), 1.00 (3H, t, J )
7.5 Hz), 0.79 (3H, t, J ) 7.2 Hz); 13C NMR (75 MHz, CDCl3) δ 174.7,
171.5, 171.0, 158.8, 158.0, 153.5, 139.8, 135.1, 131.0, 130.6 (2 C),
130.0, 129.5, 127.4, 124.6, 123.9, 123.9, 123.7, 122.4, 122.3, 119.0,
118.4, 117.2, 114.4 (2 C), 110.6, 95.6, 82.8, 76.6, 75.7, 65.9, 56.0,
55.6, 55.4, 54.6, 52.7, 52.5, 52.1, 50.1 (2 C), 45.9, 45.5, 43.2, 42.8,
40.9, 34.4, 32.8, 31.5, 27.9, 25.6, 21.1, 12.4, 8.4; ESIMS m/z 898.5
[M + 1]+; HRESIMS m/z C53H64N5O8 [M + H]+ calcd for 898.4755,
found 898.4752.
3-Demethoxycarbonyl-3-(4′-chlorobenzoylamino)methylanhy-
1
drovinblastine (18b): [R]20 +64.9 (c 0.30, CHCl3); H NMR (300
D
MHz, CDCl3) δ 9.48 (1H, s), 8.04 (1H, s), 7.73 (2H, d, J ) 8.7 Hz),
7.52 (1H, d, J ) 7.5 Hz), 7.39 (1H, d, J ) 8.7 Hz), 7.13 (3H, m), 6.93
(1H, d, J ) 6.9 Hz), 6.63 (1H, s), 6.21 (1H, s), 5.89 (1H, dd, J ) 10.2,
4.2 Hz), 5.46 (1H, s), 5.46 (1H, d, J ) 10.2 Hz), 5.09 (1H, s), 3.91
(1H, m), 3.83 (3H, s), 3.62 (3H, s), 3.40 (1H, s), 2.91 (3H, s), 2.84
(2H, d, J ) 16.5 Hz), 2.65 (1H, s), 2.12 (3H, s), 1.93 (2H, q, J ) 7.2
Hz), 1.51 (1H, m), 1.28 (1H, m), 0.99 (3H, t, J ) 7.5 Hz), 0.82 (3H,
t, J ) 7.2 Hz); 13C NMR (75 MHz, CDCl3) δ 174.9, 171.0, 166.5,
158.1, 153.6, 140.0, 137.8, 135.2, 131.2, 130.1, 130.1, 129.6, 129.0 (2
C), 128.5 (2 C), 124.7, 124.0, 123.8, 123.7, 122.5, 122.1, 119.1, 118.6,
117.5, 110.7, 95.4, 82.9, 77.0, 76.1, 66.0, 56.0, 55.7, 54.7, 52.9, 52.6,
52.3, 50.3 (2 C), 46.0, 45.4, 43.9, 42.9, 41.1, 34.5, 33.0, 31.6, 28.0,
25.8, 21.1, 12.4, 8.4; ESIMS m/z 902.5 [M + 1]+; HRESIMS m/z
C52H61ClN5O7 [M + H]+ calcd for 902.4260, found 902.4263.
3-Demethoxycarbonyl-3-(phenylacetylamino)methylanhy-
1
drovinblastine (23b): [R]20 +58.7 (c 0.15, CHCl3); H NMR (300
D
MHz, CDCl3) δ 9.25 (1H, s), 7.99 (1H, s), 7.52 (1H, d, J ) 7.5 Hz),
7.37-7.27 (5H, m), 7.19-7.11 (3H, m), 6.59 (1H, s), 6.18 (1H, d, J )
8.7 Hz), 6.09 (1H, s), 5.84 (1H, dd, J ) 10.2, 4.2 Hz), 5.45 (1H, s),
5.38 (1H, d, J ) 10.2 Hz), 4.95 (1H, s), 3.84 (3H, s), 3.79-3.77 (1H,
m), 3.63 (3H, s), 3.56 (2H, s), 3.10 (1H, s), 2.91 (1H, d, J ) 13.8 Hz),
2.80 (1H, d, J ) 15.9 Hz), 2.65 (3H, s), 2.59 (1H, s), 2.13 (3H, s),
1.92 (2H, q, J ) 7.5 Hz), 1.51-1.44 (1H, m), 1.25-1.20 (1H, m),
1.00 (3H, t, J ) 7.5 Hz), 0.79 (3H, t, J ) 7.2 Hz); 13C NMR (75 MHz,
CDCl3) δ 174.7, 171.1, 171.0, 158.0, 153.7, 135.5, 135.1, 131.0, 130.0,
129.6 (2 C), 129.4, 129.0 (2 C), 127.2, 126.3, 124.7, 124.0, 124.0,
123.7, 122.7, 121.8, 119.3, 118.4, 116.8, 110.7, 95.6, 82.7, 76.8, 75.8,
66.0, 56.0, 55.6, 54.6, 52.7, 52.6, 51.8, 50.1 (2 C), 45.8, 45.5, 44.2,
43.3, 42.9, 40.9, 34.4, 32.4, 31.5, 27.9, 24.8, 21.1, 12.4, 8.4; ESIMS
m/z 882.5 [M + 1]+; HRESIMS m/z C53H64N5O7 [M + H]+ calcd for
882.4806, found 882.4805.
3-Demethoxycarbonyl-3-(pyridine-4′-carbonylamino)methylanhy-
1
drovinblastine (19b): [R]20 +65.0 (c 0.30, CHCl3); H NMR (300
D
MHz, CDCl3) δ 9.57 (1H, s), 8.73 (2H, d, J ) 6.0 Hz), 8.03 (1H, s),
7.62 (2H, d, J ) 6.0 Hz), 7.52 (1H, d, J ) 7.5 Hz), 7.20-7.09 (3H,
m), 6.63 (1H, s), 6.22 (1H, s), 5.90 (1H, dd, J ) 10.2, 3.9 Hz), 5.46
(1H, d, J ) 10.2 Hz), 5.43 (1H, s), 5.09 (1H, s), 3.97-3.90 (1H, m),
3.83 (3H, s), 3.63 (3H, s), 3.52 (1H, d, J ) 16.5 Hz), 3.38 (1H, s),
2.91 (3H, s), 2.67 (1H, s), 2.13 (3H, s), 1.92 (2H, q, J ) 7.5 Hz),
1.53-1.48 (1H, m), 0.99 (3H, t, J ) 7.5 Hz), 0.82 (3H, t, J ) 7.2 Hz);
13C NMR (75 MHz, CDCl3) δ 174.7, 170.9, 165.4, 158.0, 153.4, 150.6
(2 C), 141.7, 140.0, 135.1, 130.9, 129.9, 129.5, 124.7, 123.8, 123.7,
123.5, 122.5, 122.1, 120.9 (2 C), 119.0, 118.5, 117.3, 110.6, 95.3, 82.8,
76.8, 75.9, 65.8, 56.0, 55.6, 54.6, 52.9, 52.5, 52.0, 50.2, 50.0, 45.8,
45.3, 43.8, 42.8, 41.2, 34.3, 32.7, 31.5, 27.9, 25.4, 21.1, 12.3, 8.4;
ESIMS m/z 869.5 [M + 1]+; HRESIMS m/z C51H61N6O7 [M + H]+
calcd for 869.4602, found 869.4601.
3-Demethoxycarbonyl-3-(3′,4′-methylenedioxybenzoylamino)me-
thylanhydrovinblastine (24b): [R]20D +69.1 (c 0.20, CHCl3); 1H NMR
(300 MHz, CDCl3) δ 9.45 (1H, s), 8.03 (1H, s), 7.53 (1H, d, J ) 8.1
Hz), 7.32 (1H, d, J ) 8.1 Hz), 7.29 (1H, s), 7.18-7.09 (3H, m), 6.83
(2H, m), 6.62 (1H, s), 6.20 (1H, s), 6.02 (2H, s), 5.90 (1H, dd, J )
10.2, 3.9 Hz), 5.46 (2H, m), 5.08 (1H, s), 3.93-3.86 (1H, m), 3.83
(3H, s), 3.63 (3H, s), 3.52 (1H, d, J ) 16.5 Hz), 3.40 (1H, s), 2.91
(3H, s), 2.65 (1H, s), 2.58 (1H, d, J ) 12.9 Hz), 2.14 (3H, s), 1.97-1.89
(2H, m), 1.54-1.47 (1H, m), 1.31-1.25 (1H, m), 0.99 (3H, t, J ) 7.5
Hz), 0.82 (3H, t, J ) 7.2 Hz); 13C NMR (75 MHz, CDCl3) δ 174.6,
3-Demethoxycarbonyl-3-(4′-nitrobenzoylamino)methylanhy-
1
drovinblastine (20b): [R]20 +64.3 (c 0.20, CHCl3); H NMR (300
D
MHz, CDCl3) δ 9.58 (1H, s), 8.28 (2H, d, J ) 8.7 Hz), 8.04 (1H, s),

Cytotoxic AnhydroVinblastine Amide DeriVatiVes
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170.8, 166.7, 158.0, 153.6, 150.3, 148.0, 139.8, 135.0, 131.0, 130.0,
129.4, 129.0, 124.7, 123.9, 123.7, 123.6, 122.9, 122.6, 121.6, 119.3,
118.5, 117.3, 110.6, 108.2, 107.7, 101.7, 95.2, 82.6, 76.9, 76.0, 66.0,
55.9, 55.5, 54.7, 52.8, 52.5, 51.7, 50.2 (2 C), 45.7, 45.4, 43.6, 42.8,
40.8, 34.4, 32.4, 31.5, 27.8, 24.9, 21.1, 12.3, 8.5; ESIMS m/z 912.4
[M + 1]+; HRESIMS m/z C53H62N5O9 [M + H]+ calcd for 912.4548,
found 912.4551.
General Procedure for the Preparation of 5b-24b Tartrate. To a
solution of tartaric acid (150 mg, 1 mmol) in acetone (20 mL) was
added dropwise a solution of compounds 5b-24b (0.5 mmol) in acetone
(10 mL). After 10 min of stirring at room temperature, the reaction
mixture was concentrated at reduced pressure to afford each compound
(5b-24b) tartrate as a white solid.
Cytotoxicity Assays. The cytotoxicity testing was carried out
according to a previously reported procedure.15
Antitumor Activity against Scarcoma 180 in Vivo. The antitumor
activity evaluation was carried out according to a previously reported
procedure.18
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Stanford, K. M.; Leamon, C. P. Bioorg. Med. Chem. Lett. 2006, 16,
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R. A.; Cullinan, G. J.; Gerzon, K.; Poore, G. A. J. Med. Chem. 1979,
22, 391–400.
Acknowledgment. This work was supported by the Chinese National
High-Tech R&D Program (grants 2007AA02Z147, 2006AA020602),
the Natural Science Foundation of China (grants 90713046, 30772638),
and the CAS Foundation (grant KSCX2-YW-R-18).
(14) Shao, Y.; Ding, H.; Tang, W. D.; Lou, L. G.; Hu, L. H. Bioorg. Med.
Chem. 2007, 15, 5061–5075.
(15) Zhang, H. K.; Shao, Y.; Ding, H.; Hu, L. H. J. Nat. Prod. 2008, 71,
1669–1676.
Supporting Information Available: NMR spectra for compounds
5b-24b. This material is available free of charge via the Internet at
http://pubs.acs.org.
(16) Sundberg, R. J.; Hong, J.; Smith, S. Q.; Sabat, M. Tetrahedron 1998,
54, 6259–6292.
References and Notes
(17) Skehan, P.; Storeng, R.; Scudiero, D.; Monks, A.; McMahon, J.;
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J. Natl. Cancer Inst. 1990, 82, 1107–1112.
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NP900157T