Tuning the acyclic ether moiety of anticancer agent AA005 with conformationally constrained fragments

Article abstract of DOI:10.1002/chem.200801298

A new series of anticancer annonaceous acetogenin mimetics were designed, synthesized, and evaluated based on our previously developed compound AA005, in which a variety of conformationally constrained fragments were introduced. Parallel syntheses of all new compounds were accomplished by replacement of the acyclic bis-ether functionality of AA005 with certain conformationally constrained fragments. Slight effects to the anticancer activity were exerted by alteringstereochemistries in the middle modification region. Similar to AA005, most newly synthesized mimetics were found to exhibit potent activities against breast cancer cells, and showed satisfactory selectivities between cancerous and non-cancerous cells. An N,N′-di-methyl bis-amide compound 67 exhibits 30 times more potency against MDA-MB-468 cells than its parent molecule AA005. This study indicates that the introduction of appropriate conformational constraints is a useful optimizing tool for this class of anticancer agents. Successes in the bisamide analogues of AA005 make this unique class of anticancer agents much simpler and more flexible for future further developments.

Full text of DOI:10.1002/chem.200801298

DOI: 10.1002/chem.200801298
Tuning the Acyclic Ether Moiety of Anticancer Agent AA005 with
Conformationally Constrained Fragments
Hai-Xia Liu, Fei Shao, Gang-Qin Li, Guo-Liang Xun, and Zhu-Jun Yao*[a]
Abstract: A new series of anticancer
annonaceous acetogenin mimetics were
designed, synthesized, and evaluated
based on our previously developed
compound AA005, in which a variety
of conformationally constrained frag-
ments were introduced. Parallel synthe-
ses of all new compounds were accom-
plished by replacement of the acyclic
bis-ether functionality of AA005 with
certain conformationally constrained
fragments. Slight effects to the anti-
cancer activity were exerted by altering
stereochemistries in the middle modifi-
cation region. Similar to AA005, most
newly synthesized mimetics were found
to exhibit potent activities against
breast cancer cells, and showed satis-
factory selectivities between cancerous
and non-cancerous cells. An N,N’-di-
methyl bis-amide compound 67 exhib-
its 30 times more potency against
MDA-MB-468 cells than its parent
molecule AA005. This study indicates
that the introduction of appropriate
conformational constraints is a useful
optimizing tool for this class of anti-
cancer agents. Successes in the bis-
amide analogues of AA005 make this
unique class of anticancer agents much
simpler and more flexible for future
further developments.
Keywords: anticancer
conformational constraints
mimicry organic chemistry
parallel synthesis
agents
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·
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Introduction
cin (a representative of this family) were replaced by an eth-
ylene glycol ether unit. These studies finally led to our in-
vention of AA005 (Scheme 1). This bis-ether compound not
only shows potent antitumor activities against a variety of
human cancer cells in the 50–100 nm range, but also exhibits
significant selectivities between a number of human normal
and cancerous cells. To generate further improved analogues
of AA005, a parallel fragment-assembly strategy was thus
developed in a systematic fashion.[6] However, all our previ-
ous works used the same protocol, in which the conforma-
tionally constrained bis-THF rings of natural acetogenin
were simply replaced by linear and rotation-free ethylene-
glycol-ether functionalities. Little is known about the confor-
mational contributions to the bioactivity when changing the
THF rings of natural products to the ethylene glycol ethers
of AA005.
To explore the above, a number of new AA005-like mole-
cules with various conformational constraints were designed
(Scheme 1). The structure of AA005 indicated that incorpo-
ration of conformationally constrained fragments into the
middle ether-bond region of AA005 is feasible by our previ-
ously developed strategy.[6] Among these, one series contain
a 1,2-disubstituted ethylene glycol unit and exist in either
the linear (e.g. compounds 1, 2,[7] Scheme 1) or cyclic state
(e.g. compounds 3–6). The other series are equipped with a
bis-amide moiety to alter the corresponding ether function-
Annonaceous acetogenins are a large family of fatty acid de-
rived natural products that feature an a,b-unsaturated g-
methylbutenolide at one terminal, one to three THF rings in
the middle region, and a hydrocarbon chain between these
two functional parts, as well as another hydrocarbon chain
at the other terminal.[1,2] Many members of this family ex-
hibit a broad spectrum of biological activities, among which
the most impressive is their anticancer activity (cytotoxic-
ity). They were considered to be the most powerful inhibi-
tors of complex I (NADH: ubiquinone oxidoreductase) in
mitochondria.[3] Due to their potent bioactivities and unique
structures, we have been engaged in modifying natural ace-
togenins (e.g. bullatacin) into the corresponding mimetics
with simpler structures for several years. In our previous
successful studies,[4,5] both the THF rings of natural bullata-
[a]Dr. H.-X. Liu, F. Shao, G.-Q. Li, Dr. G.-L. Xun, Prof. Dr. Z.-J. Yao
State Key Laboratory of Bioorganic and Natural Products Chemistry
Shanghai Institute of Organic Chemistry
Chinese Academy of Sciences, 354 Fenglin Road
Shanghai 200032 (China)
Fax : (+86)21-6416-6128
E-mail: yaoz@mail.sioc.ac.cn
Supporting information for this article is available on the WWW
under http://dx.doi.org/10.1002/chem.200801298.
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Chem. Eur. J. 2008, 14, 8632 – 8639

FULL PAPER
ieved with tosyl hydrazide
based conditions. Global depro-
tection of the MOM ethers with
BF3·Et2O in Me2S[10] provided
the final products 41–46.
Accordingly, bis-amide com-
pounds were constructed with
sequential couplings of two car-
boxylic acids (47 and 52) with
certain diamine fragments. Syn-
thesis of the left carboxylic acid
47 and the right carboxylic acid
52 is shown in Scheme 3. Initial
direct oxidation of the primary
alcohol of diol 7 by using a
TEMPO-based
(TEMPO=2,2,6,6-tetramethyl-
piperidene N-oxyl) failed,
method[11]
giving very complicated results.
A two-step alternative, Dess–
Martin oxidation of 8 followed
by treatment with sodium chlor-
ite[12] afforded the desired car-
boxylic acid 47. Synthesis of the
other carboxylic acid 52 started
Scheme 1. Design of new AA005-like molecules with conformational constraints. MOM=methoxymethyl.
ality in AA005. Amides are generally more rigid in confor-
mation than the corresponding ether compounds. Their con-
formations are, therefore, more constrained at various ex-
tents. Relative to the corresponding ether bonds, formation
of the amide bonds is more efficient and convenient in the
synthesis. In addition, the further expansion of structural di-
versity can be easily achieved by simply altering the diamine
fragments. With the above advantageous considerations, sev-
eral 1,2-diamines, 1,3-diamines, and 1,2-disubstituted ethyl-
ene glycol fragments were chosen for constructing new
AA005-like molecules in this work.
from an inexpensive commercially available material, cis-
eruic acid (Scheme 3). By using a known procedure,[5] diol
48 could be prepared in a large scale. Iterative protections
of the primary alcohol and secondary alcohol of 48 afforded
the methyl ester 49. The a,b-unsaturated lactone moiety was
constructed by a three-step sequence. Aldol condensation of
compound 49 with O-THP-(S)-lactadehyde, hydrolysis of O-
THP protecting groups, and in situ lactonization, followed
by b-elimination gave compound 50. Removal of the
TBDPS group with HF/pyridine 1:1 followed by a two-step
oxidation afforded the second acid 52.
Sequential assembly of the corresponding fragments into
the final products was carried out in parallel under standard
amide-formation conditions (EDCI and HOBt) to give dia-
mides 59–64 (Scheme 4). The final global deprotection of
the MOM ethers of 59–64 was performed in a dichlorome-
thane solution containing 30% trifluoroacetic acid and af-
forded the targets 65–70 in satisfactory yields.
Results and Discussion
Chemical synthesis: Synthesis of the first series of com-
pounds with an ether linkage is depicted in Scheme 2. Selec-
tive protection of the secondary alcohol of known diol 7[4]
afforded the corresponding MOM ether 8, the remaining
primary alcohol of which was then converted to the mesy-
late 9. Successive O-alkylation of diols 1–6 with mesylate 9
followed by (À)-(R)-epichlorohydrin[8] afforded the building
blocks 16–21 (in parallel). Regioselective opening of 16–21
by the lithium salt of trimethylsilyl acetylene in the presence
of BF3·Et2O[9] followed by treatment with MOMCl and re-
moval of TMS with TBAF afforded compounds 22–27. De-
protonation of the terminal alkynes 22–27 with nBuLi fol-
lowed by treatment with epoxide 28 in the presence of
BF3·Et2O at À788C gave the whole skeleton precursors 29–
34. Elimination of the newly born hydroxyl group with
MsCl, Et3N, and DBU afforded compounds 35–40. Selective
reduction of the middle triple and double bonds was ach-
Biological evaluation: The preliminary screening of all
newly synthesized AA005 analogues revealed that most
compounds exhibited comparable or more active potencies
than their parent AA005 against a number of cancer cell
lines. Similar to AA005, they are inactive to human normal
hepatic cell line HL7702. Such cell-type selectivity has been
observed in our previous studies on AA005 and (4R)-hy-
droxy-AA005,[5,13] though the detailed mechanisms are still
unclear.[14] Further evaluation of their inhibitory activities
against human breast cancer cell lines MDA-MB-435 and
MDA-MB-468, and non-cancerous human mammary epithe-
lial cells (HMEC) was accomplished by WST methods, by
using AA005 as a positive reference. The inhibitory poten-
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Z.-J. Yao et al.
Scheme 3. Syntheses of acids 47 and 52: a) i) DMP, CH2Cl2; ii) NaClO2,
KH2PO4, 2-methyl-2-bulene, tBuOH/H2O 4:1, 90% over 2 steps;
b) i) TBDPSCl, imidazole, CH2Cl2; ii) MOMCl, iPr2Net, CH2Cl2, 90%
over 2 steps; c) i) LDA, O-THP-(S)-lactaldehyde, THF; ii) 10% H2SO4,
THF; ii) (CF3CO)2O, Et3N, 60% over 3 steps; d) HF/pyridine 1:1, THF,
RT, 75%; e) i) DMP, CH2Cl2; ii) KH2PO4, NaClO2, 2-methyl-2-butene,
tBuOH/H2O, 86% in 2 steps. TBDPSCl=tert-butyldiphenylchlorosilane;
LDA=lithium diisopropylamide; DMP=Dess–Martin periodinane;
THP=tetrahydropyran.
Scheme 2. Syntheses of new mimetics 41–46: a) i) HC(OCH3)3, (D)-CSA,
G
CH2Cl2; ii) DIBALH in toluene, CH2Cl2, 08C, 88% over 2 steps;
b) MsCl, Et3N, CH2Cl2, 100%; c) NaH, DMF, 808C, 79–82%; d) 50%
NaOH, Bu4NHSO4, (R)-epichlorohydrin, hexane, 60–70%; e) i) nBuLi,
BF3·Et2O, trimethylsilyacetylene, THF, À788C; ii) MOMCl, iPr2NEt,
CH2Cl2; iii) TBAF, THF, 08C, 85–88% over 3 steps; f) n-BuLi, BF3·Et2O,
30, THF, À788C, 58–68%; g) MsCl, Et3N then DBU, CH2Cl2, 60–65%;
h) i) TsNHNH2, NaOAc, DME/H2O, reflux; ii) conc. HCl, MeOH, 43–
57% over 2 steps. CSA=10-camphorsulfonic acid; DIBALH=diisobutyl-
aluminum hydride; MsCl=methanesulfonyl chloride; TBAF=tetrabutyl-
ammonium fluoride; DBU=1,8-diazabicyclo[5.4.0]undec-7-ene; DME=
U
1,2-dimethoxyethane.
Scheme 4. Syntheses of amide compounds 65–70: a) EDCI, HOBt,
CH2Cl2, 85–90%; b) i) Pd/C, H2 (1 atm), MeOH; ii) 52, EDCI, HOBt, 73–
90% in 2 steps; c) 30% TFA, CH2Cl2, 57–70%. EDCI=N-(3-dimethyl-
aminopropyl)-N’-ethylcarbodiimide hydrochloride; HOBt=1-hydroxy-
benzotriazole hydrate; TFA=trifluoroacetic acid.
cies of compounds 41–46 and 65–70, in terms of IC50 values,
are listed in Table 1. All compounds except 69 and 70 show
low micromolar and submicromolar potencies against the
human breast cancer cells MDA-MB-468, whereas they are
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Anticancer Agent AA005
FULL PAPER
Table 1. Bioactivity screening of newly synthesized AA005 analogues.[a–c]
(approximately 14 times) was achieved when both amides of
65 were N-methyalted (compound 67, entry 10). A similar
but more rigid compound 68 with a 1,4-piperazine moiety
(entry 11) could not afford a better record, though it is 2–3
times more potent than amides 65 and 66. To our surprise,
introduction of 1,2-trans-cyclohexanediamine into the skele-
ton of AA005 yielded negative results (entries 12 and 13).
Generally, the amide is more rigid in conformation than the
corresponding ether, even in the acyclic states. Based on the
above results, it can be concluded that conformational con-
straint is a double-edged sword for the bioactivity of such
AA005-like molecules: An appropriate conformational con-
straint with slightly rigid properties, such as that introduced
into 67, is mostly favorable. Second, stereochemistry affects
the bioactivity of these AA005-like compounds little or to a
very limited extent. A similar phenomenon was also ob-
served in the naturally occurring annonaceous acetogenins
with the hydroxylated bis-THF ring(s), the stereochemistries
of which hardly affect the inhibitory potency.[15] In this
study, for instance, pairs of diastereomerically different (en-
antiomers, each in the modification region) compounds, 41
and 42 (entries 2 and 3), 43 and 44 (entries 4 and 5), and 45
and 46 (entries 6 and 7), exhibit almost the same potencies.
The two worst compounds in this study, 69 and 70 (en-
tries 12 and 13), gave only a five times potency difference
against MDA-MB468. In addition, the nature of heteroa-
toms (O, S, or N) introduced in these compounds is not cru-
cial to the bioactivity, though nitrogen-containing com-
pounds give relatively better results.
Entry
Compounds
IC50 [mm]
MDA-MB-435
MDA-MB-468
HMEC
1
2
3
4
5
6
7
8
AA005
41
42
43
44
45
46
65
66
67
68
69
70
>100
6.467
4.170
5.500
11.24
25.69
18.40
>100
>100
3.784
12.61
>100
>100
5.932
0.830
1.005
0.994
1.630
2.559
3.007
2.953
2.019
0.218
0.858
11.81
61.59
82.11
14.25
10.86
13.60
17.68
20.63
17.95
>100
>100
15.11
70.00
>100
>100
9
10
11
12
13
[a]AA005 was used as a positive control. [b]MDA-MB-435: human
breast cancer cell; MDA-MB-468: human breast cancer cell; HMEC:
non-cancerous human mammary epithelial cells. [c]Inhibition of cell
growth by the listed compounds in MDA-MB-435, MDA-MB-468, and
HMEC cells was determined by using a WST assay.
less active against the other type of human breast cancer
cells MDA-MB-435. Eight samples presented at least 10
times less potency, and four of them are inactive (IC50 >
100 mm) against noncancerous HMEC, when compared to
MDA-MB-468. Compound 67, the most potent compound
in this study, exhibits 69 times more potency against MDA-
MB-468 than HMEC cells.
Based on their cytotoxicities against MDA-MB-468, some
conclusions can be drawn with relevance to their structural
features. In general, all the newly synthesized compounds
are more active than their parent AA005, with the exception
of two more rigid compounds 69 and 70 (Table 1, entries 12
and 13). Among them, compound 67 (entry 10) exhibited
the highest potency with an IC50 of 0.218 mm, 30-times great-
er than that of AA005. Obviously, the introduction of ap-
propriate conformational constraints into the middle ether
region of AA005 is a useful strategy for the improvement of
anticancer activity. Furthermore, compound 67 (entry 10,
SI=69) shows a better selectivity for inhibitory effects be-
tween cancerous MDA-MB-468 cells and noncancerous
HMEC cells than its parent AA005 (entry 1, SI=14).
For the first series of compounds with ether functionalities
(Table 1, entries 2–7), either in the open or cyclic state, the
bioactivities are similar. Only limited improvements were
achieved. The best of this series are compounds 41 and 43
(IC50 ꢀ1 mm, entries 2 and 4), as both of these compounds
acquired a potency 5–6 times that of AA005. However,
much more different results were observed in the second
series, the amide-containing compounds. It was the first op-
portunity to introduce nitrogen atom(s) into the middle
region of the AA005 skeleton. Among the six nitrogen-con-
taining compounds, we can find the best (67; IC50 =0.218 mm,
entry 10) and worst compound (70; IC50 =61.59 mm,
entry 13) in this study. Linear amides 65 (with 1,2-ethanedi-
amine as the linkage) and 66 (with 1,3-propanediamine as
the linkage) slightly improved their cytotoxicities (entries 8
and 9). More interestingly, a great extent of enhancement
Conclusion
A number of new analogues based on anticancer annona-
ceous acetogenin mimetic AA005 were designed and synthe-
sized by the introduction of conformational constraints to
the linear skeleton of AA005. Biological results demonstrate
that stereochemical variations in the modification region ex-
erted little effects on the bioactivity. Most AA005 analogues
in this study were found to be more potent against breast
cancer cell line MDA-MB-468 than the other cancer cell
line MDA-MB-435 and showed satisfactory selectivity to
noncancerous cell line HMEC. The generally improved bio-
activities clearly indicate that the introduction of appropri-
ate conformational constraints into the linear AA005 skele-
ton is a useful optimizing tool for this unique class of anti-
cancer agents. In addition, success with the amide analogues
of AA005 makes the AA005-based anticancer agents much
simpler to synthesize and more flexible for future further
development.
Experimental Section
General methods: Optical rotations were measured by a Perkin–Elmer
341mC polarimeter at room temperature. IR spectra were obtained by
using a Fourier-transform IR spectrometer (FTIR). 1H NMR spectra
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Z.-J. Yao et al.
were recorded with Varian EM and Bruker AMX machines at 300, 400,
or 500 MHz and are reported in ppm (d) downfield relative to TMS as
the internal standard, and 13C NMR spectra were recorded at 100 and
150 MHz and assigned in ppm (d). HRMS spectra were recorded by
using either a Kratos Concept instrument, a Q-Tof micro instrument, or
an APEXIII 7.0 TESLA FTMS. Elemental analysis was preformed by
using an Elemental VARIO EL apparatus. All melting points were un-
corrected. Flash column chromatography was performed on silica gel
(10–40 mm).
CDCl3): d=0.87 (3H, t, J=7.2 Hz), 1.25–1.53 (30H, m), 1.40 (3H, d, J=
6.9 Hz), 2.25 (2H+1OH, m), 2.46 (4H, m), 2.83 (2H, m), 2.96 (1H, d,
J=3.9 Hz), 3.01 (1H, d, J=3.9 Hz), 3.37 (3H, s), 3.38 (3H, s), 3.45–3.69
(7H, m), 3.81 (1H, m), 4.06 (2H, m), 4.64 (1H, d, J=6.6 Hz), 4.71 (2H,
s), 4.74 (1H, d, J=6.6 Hz), 4.99 (1H, dq, J=1.5, 6.6 Hz), 6.98 ppm (1H,
d, J=1.2 Hz); 13C NMR (100 MHz, CDCl3): d=173.9, 148.9, 134.2, 96.0,
95.9, 84.6, 84.4, 78.8, 78.4, 77.4, 76.2, 74.7, 72.2, 70.6, 70.1, 55.56, 55.51,
36.3, 32.87, 32.82, 31.97, 31.92, 29.7, 29.5, 29.4, 29.3, 29.2, 29.1, 27.8, 27.3,
25.6, 25.4, 25.1, 22.7, 22.1, 19.2, 14.1 ppm; MS (ESI): m/z: 735 (M++Na);
HRMS (ESI): m/z: calcd for C39H68O9SNa: 735.4476; found: 735.4484; el-
emental analysis calcd (%) for C39H68O9S: C 65.70, H 9.61; found: C
66.00, H 9.61.
General procedure for 29–34: nBuLi (1.6m in hexanes, 2 mmol) at
À788C was added to a solution of 22–27 (2 mmol) in THF (5 mL). After
the mixture had been stirred for 30 min, BF3·Et2O (2 mmol) was added.
After an additional 30 min at À788C, a solution of 28 (2 mmol) in THF
(5 mL) was added dropwise. The reaction mixture was quenched after 2 h
by the addition of aq. NH4Cl solution and subsequently extracted with
ethyl acetate (3”25 mL). The combined organic layers were washed with
water and brine, concentrated, and then purified by column chromatogra-
phy with EtOAc/petroleum ether 1:5 to 1:2 as the eluent to give com-
pounds 29–34, respectively.
Data for 34: Yellowish oil; yield: 68%; IR (film): n˜ =3496, 2927, 2855,
1757, 1465, 1357, 1318, 1204, 1152, 1106, 1036, 918 cmÀ1 1H NMR
;
(500 MHz, CDCl3): d=0.86 (3H, t, J=7.2 Hz), 1.25–1.53 (30H, m), 1.39
(3H, d, J=6.9 Hz), 2.25 (2H+1OH, m), 2.46 (4H, m), 2.82 (2H, m),
2.97 (1H, d, J=4.2 Hz), 3.01 (1H, d, J=4.2 Hz), 3.37 (3H, s), 3.38 (3H,
s), 3.43–3.69 (7H, m), 3.80 (1H, m), 4.05 (2H, m), 4.63 (1H, d, J=
6.6 Hz), 4.71 (2H, s), 4.74 (1H, d, J=6.6 Hz), 4.99 (1H, dq, J=1.8,
6.6 Hz), 6.98 ppm (1H, d, J=1.5 Hz); 13C NMR (100 MHz, CDCl3): d=
173.9, 148.9, 134.2, 96.1, 95.9, 84.6, 84.4, 78.8, 77.4, 76.4, 74.7, 72.3, 70.7,
70.1, 55.5, 36.3, 32.8, 32.0, 31.9, 29.7, 29.5, 29.4, 29.3, 29.2, 29.1, 27.8, 27.3,
25.6, 25.3, 25.1, 22.7, 22.1, 19.2, 14.1 ppm; MS (ESI): m/z: 735 [M++Na];
HRMS (ESI): m/z: calcd for C39H68O9SNa: 735.4476; found: 735.4481; el-
emental analysis calcd (%) for C39H68O9S: C 65.70, H 9.61; found: C
65.69, H 9.85.
Data for 29: Yellowish oil; yield: 58%; IR (film): n˜ =3487, 2926, 2856,
2249, 1757, 1465, 1320, 1314, 1152, 1111, 1038, 919, 733 cmÀ1 1H NMR
;
(500 MHz, CDCl3): d=0.88 (3H, t, J=7.0 Hz), 1.26 (24H, m), 1.40 (3H,
d, J=6.8 Hz), 1.54 (6H, m), 2.26 (2H, t, J=7.3 Hz), 2.47 (4H, m), 3.36
(6H, s), 3.37 (3H, s), 3.38 (3H, s), 3.37 (1H, m), 3.56–3.85 (12H, m), 4.62
(4H, s), 4.63 (1H, d, J=6.7 Hz), 4.72 (2H, s), 4.76 (1H, d, J=6.8 Hz),
4.99 (1H, dq, J=1.5, 6.7 Hz), 6.99 ppm (1H, d, J=1.0 Hz); 13C NMR
(100 MHz, CDCl3): d=173.7, 148.8, 134.1, 96.6, 95.9, 95.8, 79.4, 79.1, 78.2,
76.4, 74.8, 73.8, 69.9, 66.9, 55.37, 55.33, 55.1, 36.2, 32.0, 31.7, 29.6, 29.5,
29.3, 29.2, 29.1, 29.0, 27.7, 27.2, 25.5, 25.3, 25.0, 22.5, 22.1, 19.0, 13.9 ppm;
MS (ESI): m/z: 825 [M++Na]; HRMS (ESI): m/z: calcd for
C43H78O13Na: 825.5334; found: 825.5362.
General procedure for compounds 35–40: MsCl (1 mmol) in CH2Cl2
(2 mL) was added to a stirred solution of 29–34 (0.5 mmol) and Et3N
(1.5 mmol) in CH2Cl2 (5 mL) at 08C. The reaction mixture was stirred at
RT for 30 min, and then DBU (1 mmol) was added. The mixture was
stirred for a further 2 h, and was then quenched with water (5 mL) and
extracted with CH2Cl2 (3”20 mL). The combined organic layers were
washed with HCl (1n), water, and brine, and then dried over Na2SO4.
Removal of the solvent followed by column chromatography with
EtOAc/petroleum ether 1:3 to 1:1 as the eluent gave compounds 35–40,
respectively.
Data for 30: Yellowish oil; yield: 58%; IR (film): n˜ =3494, 2927, 2856,
2249, 1757, 1465, 1319, 1214, 1152, 1037, 919, 756, 734 cmÀ1 1H NMR
;
(500 MHz, CDCl3): d=0.88 (3H, t, J=7.0 Hz), 1.26 (24H, m), 1.40 (3H,
d, J=6.8 Hz), 1.53 (6H, m), 2.26 (2H, t, J=7.0 Hz), 2.47 (4H, m), 3.36
(6H, s), 3.37 (3H, s), 3.38 (3H, s), 3.37 (1H, m), 3.62 (8H, m), 3.76 (4H,
m), 4.62 (4H, s), 4.64 (1H, d, J=6.8 Hz), 4.72 (2H, m), 4.77 (1H, d, J=
6.8 Hz), 4.99 (1H, dq, J=1.5, 6.8 Hz), 6.99 ppm (1H, d, J=1.3 Hz);
13C NMR (100 MHz, CDCl3): d=173.7, 148.8, 134.1, 96.6, 96.0, 95.8, 79.3,
79.1, 76.5, 74.9, 74.0, 72.3, 69.9, 66.9, 55.36, 55.33, 55.1, 36.2, 32.0, 31.7,
29.6, 29.5, 29.3, 29.2, 29.1, 28.9, 27.7, 27.2, 25.5, 25.2, 25.0, 22.5, 22.1, 19.0,
13.9 ppm; MS (ESI): m/z: 825 [M++Na]; HRMS (ESI): m/z: calcd for
C43H78O13Na: 825.5334; found: 825.5301.
Data for 35: Yellowish oil; yield: 62%; IR (film): n˜ =2928, 2856, 1759,
1465, 1319, 1214, 1152, 1110, 1039, 919 cmÀ1
;
1H NMR (400 MHz,
CDCl3): d=0.88 (3H, t, J=7.0 Hz), 1.26 (24H, m), 1.40 (3H, d, J=
6.8 Hz), 1.52 (6H, m), 2.06 (1H, m), 2.26 (3H,m), 2.62 (2H, m), 3.35
(6H, s), 3.37 (3H, s), 3.39 (3H, s), 3.53–3.87 (13H, m), 4.62 (5H, m), 4.74
(3H, m), 4.99 (1H, dq, J=1.6, 6.8 Hz), 5.43 (1H, m), 5.81 (0.55H, dt, J=
10.7, 7.3 Hz), 6.04 (0.46H, dt, J=15.8, 7.0 Hz), 6.99 ppm (1H, d, J=
1.5 Hz); MS (ESI): m/z: 807 [M++Na]; HRMS (ESI): m/z: calcd for
C43H76O12Na: 807.5229; found: 807.5254.
Data for 31: Yellowish oil; yield: 60%; IR (film): n˜ =3489, 2926, 2856,
1756, 1467, 1319, 1214, 1151, 1105, 1033, 919 cmÀ1 1H NMR (500 MHz,
;
CDCl3): d=0.88 (3H, t, J=7.0 Hz), 1.26 (24H, m), 1.40 (3H, d, J=
6.8 Hz), 1.53 (4H, m), 2.27 (2H+1OH, m), 2.45 (4H, m), 3.37 (3H, s),
3.38 (3H, s), 3.51 (2H, m), 3.67 (4H, m), 3.80 (3H, m), 3.92 (2H, m),
3.98 (2H, m), 4.64 (1H, d, J=6.8 Hz), 4.73 (3H, m), 4.99 (1H, dq, J=1.5,
6.7 Hz), 6.99 ppm (1H, s); 13C NMR (100 MHz, CDCl3): d=173.0, 148.1,
133.4, 95.2, 95.1, 82.8, 77.7, 75.4, 73.8, 71.4, 70.6, 70.4, 69.7, 69.3, 54.7,
54.6, 35.5, 31.2, 31.1, 28.9, 28.8, 28.6, 28.5, 28.4, 28.3, 27.0, 26.6, 24.8, 24.6,
24.3, 21.8, 21.3, 18.4, 13.3 ppm; MS (ESI): m/z: 719 [M++Na]; HRMS
(ESI): m/z: calcd for C39H68O10Na: 719.4704; found: 719.4726.
Data for 36: Yellowish oil; yield: 62%; IR (film): n˜ =2928, 2856, 1759,
1465, 1401, 1319, 1214, 1153, 1111, 954, 919 cmÀ1 1H NMR (400 MHz,
;
CDCl3): d=0.88 (3H, t, J=7.0 Hz), 1.26 (24H, m), 1.40 (3H, d, J=
6.8 Hz), 1.52 (6H, m), 2.06 (1H, m), 2.26 (3H,m), 2.62 (2H, m), 3.36
(6H, s), 3.37 (3H, s), 3.39 (3H, s), 3.37 (2H, m), 3.53–3.87 (11H, m), 4.63
(5H, m), 4.74 (3H, m), 4.99 (1H, dq, J=1.6, 6.8 Hz), 5.43 (1H, m), 5.81
(0.55H, dt, J=10.7, 7.3 Hz), 6.04 (0.46H, dt, J=15.8, 7.0 Hz), 6.99 ppm
(1H, d, J=1.5 Hz); MS (ESI): m/z: 807 [M++Na]; HRMS (ESI): m/z:
calcd for C43H76O12Na: 807.5229; found 807.5233.
Data for 32: Yellowish oil; yield: 60%; IR (film): n˜ =3489, 2927, 2856,
Data for 37: Yellowish oil; yield: 60%; IR (film): 3489, 2926, 2856, 1756,
1757, 1465, 1319, 1213, 1152, 1106, 1035, 919 cmÀ1
;
1H NMR (500 MHz,
1467, 1319, 1214, 1151, 1105, 1033, 919 cmÀ1 1H NMR (500 MHz,
;
CDCl3): d=0.88 (3H, t, J=7.0 Hz), 1.26 (24H, m), 1.40 (3H, d, J=
6.8 Hz), 1.53 (4H, m), 2.27 (2H+1OH, m), 2.46 (4H, m), 3.38 (3H, s),
3.39 (3H, s), 3.50 (2H, m), 3.62 (2H, m), 3.67 (2H, m), 3.81 (3H, m),
3.94 (4H, m), 4.65 (1H, d, J=6.8 Hz), 4.72 (3H, m), 4.99 (1H, dq, J=1.6,
6.8 Hz), 7.00 ppm (1H, d, J=1.4 Hz); 13C NMR (100 MHz, CDCl3): d=
173.0, 148.1, 133.4, 95.3, 95.1, 82.8, 82.7, 77.8, 75.5, 73.9, 71.5, 70.6, 70.5,
69.8, 69.2, 54.7, 54.6, 35.5, 31.2, 31.0, 28.8, 28.7, 28.6, 28.5, 28.4, 28.2, 27.0,
26.5, 24.8, 24.6, 24.3, 21.8, 21.2, 18.3, 13.2 ppm; MS (ESI): m/z: 719
[M++Na]; HRMS (ESI): m/z: calcd for C39H68O10Na: 719.4704; found:
719.4733.
CDCl3): d=0.88 (3H, t, J=7.0 Hz), 1.26 (24H, m), 1.40 (3H, d, J=
6.8 Hz), 1.53 (4H, m), 2.27 (2H+1OH, m), 2.45 (4H, m), 3.37 (3H, s),
3.38 (3H, s), 3.51 (2H, m), 3.67 (4H, m), 3.80 (3H, m), 3.92 (2H, m),
3.98 (2H, m), 4.64 (1H, d, J=6.8 Hz), 4.73 (3H, m), 4.99 (1H, dq, J=1.5,
6.7 Hz), 6.99 ppm (1H, s); MS (ESI): m/z: 719 [M++Na]; HRMS (ESI):
m/z: calcd for C39H68O10Na: 719.4704; found: 719.4726.
Data for 38: Yellowish oil; yield: 60%; IR (film): n˜ =2927, 2855, 1758,
1464, 1356, 1318, 1216, 1152, 1105, 1037, 918 cmÀ1 1H NMR (500 MHz,
;
CDCl3): d=0.88 (3H, t, J=7.1 Hz), 1.26 (24H, m), 1.40 (3H, d, J=
6.8 Hz), 1.53 (4H, m), 2.06 (1H, m), 2.26 (3H, m), 2.61 (2H, m), 3.37
(3H, s), 3.39 (3H, s), 3.49 (2H, m), 3.78 (2H, m), 3.85 (1H, m), 3.94
(4H, m), 4.64 (1H, d, J=6.8 Hz), 4.73 (3H, m), 5.00 (1H, dq, J=1.5,
Data for 33: Yellowish oil; yield: 67%; IR (film): n˜ =3489, 2927, 2855,
1756, 1465, 1318, 1204, 1152, 1105, 1036„ 918 cmÀ1 1H NMR (500 MHz,
;
8636
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Chem. Eur. J. 2008, 14, 8632 – 8639

Anticancer Agent AA005
FULL PAPER
6.8 Hz), 5.43 (1H, m), 5.83 (0.55H, dt, J=10.6, 7.4 Hz), 6.05 (0.45H, dt,
J=15.2, 7.4 Hz), 6.99 ppm (1H, s); MS (ESI): m/z: 701 [M++Na];
HRMS (ESI): m/z: calcd for C39H66O9Na: 701.4599; found: 701.4588.
25.4, 25.1, 22.6, 19.1, 14.0 ppm; MS (ESI): m/z: 619 [M++Na]; HRMS
(ESI): m/z: calcd for C35H64O7Na: 619.4568; found: 619.4524.
Data for 44: White wax; yield: 57%; IR (film): n˜ =3500, 2920, 2851,
1743, 1467, 1325, 1147, 1074, 1031, 953, 727 cmÀ1; [a]D25 =+0.14 (c=0.8 in
CHCl3); 1H NMR (500 MHz, CDCl3): d=0.88 (3H, t, J=7.1 Hz), 1.26
(32H, m), 1.40 (3H, d, J=6.8 Hz), 1.44 (6H, m), 1.54 (2H, m), 2.26 (2H,
t, J=7.7 Hz), 2.30 (2OH, brs), 3.33 (2H, m), 3.51 (2H, m), 3.77 (4H, m),
3.94 (2H, m), 3.99 (2H, m), 4.99 (1H, dq, J=1.5, 6.8 Hz), 6.99 ppm (1H,
J=1.2 Hz); 13C NMR (100 MHz, CDCl3): d=173.8, 148.8, 134.3, 83.4,
77.3, 73.9, 71.4, 70.3, 33.0, 31.8, 29.6, 29.5, 29.2, 29.1, 27.3, 25.4, 25.1, 22.6,
19.1, 14.0 ppm; MS (ESI): m/z: 619 [M++Na]; HRMS (ESI): calcd for
C35H64O7Na: 619.4568; found: 619.4562.
Data for 39: Yellowish oil; yield: 65%; IR (film): n˜ =2926, 2854, 1758,
1465, 1318, 1152, 1105, 1037, 918 cmÀ1 1H NMR (500 MHz, CDCl3): d=
;
0.88 (3H, t, J=7.2 Hz), 1.26–1.53 (30H, m), 1.41 (3H, d, J=6.9 Hz), 2.07
(1H, m), 2.26 (2H, t, J=7.5 Hz), 2.26 (1H, m), 2.58 (1H, m), 2.64 (1H,
m), 2.86 (2H, m), 2.99 (1H, d, J=3.6 Hz), 3.02 (1H, d, J=6.6 Hz), 3.38
(3H, s), 3.39 (3H, s), 3.46–3.71 (5H, m), 3.85 (1H, m), 4.08 (2H, m), 4.65
(1H, d, J=6.6 Hz), 4.73 (2H, s), 4.74 (1H, d, J=6.6 Hz), 4.99 (1H, dq,
J=1.8, 6.6 Hz), 5.44 (1H, m), 5.83 (0.55H, dt, J=11.1, 7.5 Hz), 6.05
(0.45H, dt, J=15.9, 6.9 Hz), 6.99 ppm (1H, d, J=1.5 Hz); MS (ESI):
m/z: 717 [M++Na]; HRMS (ESI): m/z: calcd for C39H66O8SNa: 717.4370;
found: 717.4399.
Data for 45: White wax; yield: 46%; [a]2D5 =+38.1 (c=2.0 in CHCl3); IR
(film): n˜ =3498, 2926, 2850, 1745, 1468, 1324, 1166, 1121, 1029, 953, 863,
721 cmÀ1 1H NMR (500 MHz, CDCl3): d=0.88 (3H, t, J=7.1 Hz), 1.26–
;
Data for 40: Yellowish oil; yield: 65%; IR (film): n˜ =2926, 2854, 1758,
1465, 1318, 1152, 1105, 1037, 918 cmÀ1 1H NMR (500 MHz, CDCl3): d=
;
1.56 (40H, m), 1.41 (3H, d, J=6.8 Hz), 2.26 (2H, t, J=7.9 Hz), 2.47
(2OH, brs), 2.84 (2H, m), 3.00 (2H, m), 3.36 (2H, dd, J=7.9, 9.4 Hz),
3.53 (2H, dd, J=2.8, 9.6 Hz), 3.74 (2H, m), 4.08 (2H, m), 4.99 (1H, dq,
J=1.4, 6.6 Hz), 6.99 ppm (1H, d, J=1.4 Hz); 13C NMR (100 MHz,
CDCl3): d=173.8, 148.8, 134.2, 83.9, 77.3, 73.8, 70.1, 33.0, 32.9, 32.2, 31.8,
29.6, 29.4, 29.2, 29.1, 27.3, 25.4, 25.1, 22.6, 19.1, 14.0 ppm; MS (ESI): m/z:
635 [M++Na]; HRMS (ESI): m/z: calcd for C35H64O6SNa: 635.4315;
found: 635.4331.
0.88 (3H, t, J=7.2 Hz), 1.25–1.53 (30H, m), 1.41 (3H, d, J=6.9 Hz), 2.07
(1H, m), 2.26 (2H, t, J=7.5 Hz), 2.26 (1H, m), 2.58 (1H, m), 2.64 (1H,
m), 2.82 (1H, dd, J=3.3, 6.3 Hz), 2.87 (1H, dd, J=3.3, 5.4 Hz), 2.99 (1H,
d, J=4.2 Hz), 3.02 (1H, J=4.2 Hz), 3.38 (3H, s), 3.39 (3H, s), 3.44–3.71
(5H, m), 3.84 (1H, m), 4.08 (2H, m), 4.65 (1H, d, J=6.6 Hz), 4.73 (2H,
s), 4.74 (1H, d, J=6.6 Hz), 4.99 (1H, dq, J=1.5, 6.6 Hz), 5.45 (1H, m),
5.83 (0.55H, dt, J=11.1, 7.5 Hz), 6.06 (0.45H, dt, J=14.4, 7.8 Hz),
6.99 ppm (1H, d, J=1.5 Hz); MS (ESI): m/z: 717 [M++Na]; HRMS
(ESI): m/z: calcd for C39H66O8SNa: 717.4370; found: 717.4382.
Data for 46: White wax; yield: 43%; IR (film): n˜ =3463, 2924, 2851,
1750, 1467, 1320, 1175, 1087, 1027, 948, 869, 722 cmÀ1; [a]D25 =À29.0 (c=
1.55 in CHCl3); 1H NMR (500 MHz, CDCl3): d=0.88 (3H, t, J=7.0 Hz),
1.26–1.56 (40H, m), 1.41 (3H, d, J=6.8 Hz), 2.26 (2H, t, J=7.7 Hz), 2.83
(2H, m), 2.99 (1H, d, J=3.4 Hz), 3.02 (1H, d, J=3.6 Hz), 3.33 (2H, dd,
J=8.5, 8.5 Hz), 3.59 (2H, dd, J=2.9, 9.5 Hz), 3.74 (2H, m), 4.09 (2H, m),
4.99 (1H, q, J=6.6 Hz), 6.99 ppm (1H, s); 13C NMR (100 MHz, CDCl3):
d=173.7, 148.8, 134.2, 84.5, 77.3, 74.3, 70.5, 33.0, 32.9, 32.0, 31.8, 29.5,
29.4, 29.2, 29.1, 27.3, 25.4, 25.1, 22.6, 19.1, 14.0 ppm; MS (ESI): m/z: 635
[M++Na]; HRMS (ESI): m/z: calcd for C35H64O6SNa: 635.4315; found:
635.4332.
General procedure for compounds 41–46: NaOAc (22.0 mmol) in water
(30 mL) was added over 5 h to a stirred solution of 35–40 (0.15 mmol)
and p-toluenesulfonyl hydrazide (18.6 mmol) in dimethoxyethane
(25 mL) under reflux. The mixture was then cooled down to RT, poured
into water, and extracted with ether (3”20 mL). The combined organic
layers were dried over Na2SO4 and concentrated under reduced pressure.
The residue was directly used in the next step without further purifica-
tion. Concentrated HCl (0.5 mL) was added to a stirred solution of the
above crude product in methanol (2 mL) at 08C. Then, the reaction mix-
ture was warmed to RT and stirred for 24 h. Finally, the mixture was
quenched with sat. NaHCO3 at 08C. The mixture was extracted with
ethyl acetate (3”15 mL) and the extracts were washed with brine, dried
over Na2SO4, and concentrated under reduced pressure. The residue was
purified by column chromatography with EtOAc/petroleum ether 1:5 to
1:1 as the eluent to afford compounds 41–46, respectively.
General procedure for 59–64: Compounds 53–58 (1 mmol) in methanol
(10 mL) were hydrogenated under H2 (1 atm) in the presence of 10%
Pd/C at room temperature for 4 h. After removal of solid through Celite
and evaporation of solvents, the crude product was directly used in the
next step. The mixture of this crude product, compound 52 (0.19 g,
0.5 mmol), and HOBt (0.13 g, 1.0 mmol) in dry CH2Cl2 (15 mL) was
treated with EDCI (0.19 g, 1.0 mmol) at 08C. The reaction mixture was
stirred for 4 h at RT, quenched with saturated NH4Cl, and extracted with
ethyl acetate. The extracts were washed with saturated NaHCO3 and
brine, dried over Na2SO4, and concentrated under reduced pressure. The
residue was purified by column chromatography with EtOAc/petroleum
ether 1:5 to 1:1 as the eluent to give compounds 59–64, respectively.
Data for 41: White wax; yield: 46%; [a]2D5 =À16.1 (c=0.6 in CHCl3); IR
(film): n˜ =3353, 2925, 2851, 1749, 1471, 1326, 1201, 1144, 1113, 1033, 880,
720 cmÀ1 1H NMR (400 MHz, CDCl3): d=0.88 (3H, t, J=7.0 Hz), 1.26–
;
1.50 (38H, m), 1.40 (3H, d, J=6.8 Hz), 1.54 (2H, m), 2.26 (2H, t, J=
8.0 Hz), 3.18 (4OH, brs), 3.44 (2H, m), 3.65 (6H, m), 3.79 (4H, m), 4.99
(1H, dq, J=1.6, 6.8 Hz), 6.99 ppm (1H, d, J=1.5 Hz); 13C NMR
(100 MHz, CDCl3): d=173.9, 148.9, 134.3, 80.6, 77.4, 75.2, 70.7, 60.5, 33.1,
31.9, 29.7, 29.68, 29.63, 29.5, 29.4, 29.3, 29.2, 29.1, 27.4, 25.5, 25.1, 22.6,
19.1, 14.0 ppm; MS (ESI): m/z: 637 [M++Na]; HRMS (ESI): m/z: calcd
for C35H66O8Na: 637.4649; found: 637.4678.
Data for 59: White solid; yield: 77%; [a]2D5 =À37.8 (c=1.90 in CHCl3);
IR (film): n˜ =3291, 3077, 2920, 2851, 1745, 1660, 1547, 1471, 1382, 1323,
1233, 1161, 1104, 1051, 1036, 958, 920, 719 cmÀ1 1H NMR (500 MHz,
;
Data for 42: White wax; yield: 54%; [a]2D5 =À0.11 (c=1.45 in CHCl3);
CDCl3): d=0.88 (3H, t, J=7.1 Hz), 1.25 (34H, m), 1.40 (3H, d, J=
6.8 Hz), 1.54 (2H, m), 1.75 (4H, m), 2.26 (2H, t, J=8.0 Hz), 3.39 (6H, s),
3.47 (4H, m), 4.03 (2H, m), 4.62 (2H, d, J=6.6 Hz), 4.65 (2H, d, J=
6.6 Hz), 4.99 (1H, q, J=6.8 Hz), 6.99 (1H, s), 7.02 ppm (2H, brs);
13C NMR (125 MHz, CDCl3): d=173.8, 173.5, 148.8, 134.3, 96.2, 77.9,
77.3, 56.0, 39.2, 32.9, 31.8, 29.5, 29.4, 29.3, 29.2, 29.1, 27.3, 25.1, 24.8, 22.6,
19.1, 14.0 ppm; MS (ESI): m/z: 691 [M++Na]; HRMS (MALDI): m/z:
calcd for C37H68N2O8Na: 691.4867; found: 691.4854.
IR (film): n˜ =3356, 2920, 2851, 1753, 1469, 1321, 1084, 1029, 889,
721 cmÀ1 1H NMR (400 MHz, CDCl3): d=0.88 (3H, t, J=7.0 Hz), 1.26–
;
1.50 (38H, m), 1.40 (3H, d, J=6.8 Hz), 1.54 (2H, m), 2.26 (2H, t, J=
8.0 Hz), 3.39 (2H, m), 3.44 (4OH, brs), 3.75 (10H, m), 4.99 (1H, dq, J=
1.6, 6.8 Hz), 6.99 ppm (1H, d, J=1.5 Hz); 13C NMR (100 MHz, CDCl3):
d=173.9, 148.9, 134.2, 80.8, 77.4, 75.5, 71.0, 60.7, 32.9, 31.8, 29.68, 29.66,
29.60, 29.5, 29.4, 29.3, 29.2, 29.1, 27.3, 25.6, 25.1, 22.6, 19.1, 14.0 ppm; MS
(ESI): m/z: 637 [M++Na]; HRMS (ESI): m/z: calcd for C35H66O8Na:
637.4649; found: 637.4676.
Data for 60: White solid; yield: 90%; [a]2D5 =À40.6 (c=2.40 in CHCl3);
IR (film): n˜ =3295, 2923, 2852, 2822, 1755, 1652, 1535, 1468, 1434, 1371,
1319, 1215, 1152, 1128, 1095, 1038, 919, 723, 670 cmÀ1
;
1H NMR
Data for 43: White wax; yield: 57%; [a]2D5 =+12.5 (c=0.95 in CHCl3);
(500 MHz, CDCl3): d=0.88 (3H, t, J=7.1 Hz), 1.25 (32H, m), 1.40 (3H,
d, J=6.8 Hz), 1.52 (2H, m), 1.73 (6H, m), 2.26 (2H, t, J=8.0 Hz), 3.30
(4H, m), 3.40 (6H, s), 4.05 (2H, m), 4.67 (2H, d, J=6.6 Hz), 4.69 (2H, d,
J=6.6 Hz), 5.00 (1H, dq, J=1.6, 6.8 Hz), 6.99 (1H, d, J=1.6 Hz),
7.06 ppm (2H, m); 13C NMR (125 MHz, CDCl3): d=173.7, 173.0, 148.8,
134.2, 96.2, 77.9, 77.2, 56.0, 35.3, 33.0, 31.8, 29.9, 29.6, 29.5, 29.4, 29.3,
29.2, 29.1, 27.3, 25.1, 24.8, 22.6, 19.1, 14.0 ppm; MS (ESI): m/z: 705 [M+
IR (film): n˜ =3500, 2919, 2850, 1747, 1467, 1323, 1138, 1122, 1079, 1027,
880, 722 cmÀ1 1H NMR (500 MHz, CDCl3): d=0.88 (3H, t, J=7.1 Hz),
;
1.26 (32H, m), 1.40 (3H, d, J=6.8 Hz), 1.44 (6H, m), 1.54 (2H, m), 2.26
(2H+2OH, m), 3.31 (2H, m), 3.52 (2H, m), 3.73 (2H, m), 3.79 (2H, m),
3.94 (2H, m), 3.99 (2H, m), 4.99 (1H, dq, J=1.6, 6.8 Hz), 6.99 ppm (1H,
J=1.5 Hz); 13C NMR (100 MHz, CDCl3): d=173.8, 148.8, 134.2, 83.4,
77.3, 73.9, 71.4, 70.3, 70.2, 33.0, 31.8, 29.6, 29.5, 29.27, 29.23, 29.1, 27.3,
Chem. Eur. J. 2008, 14, 8632 – 8639
ꢀ 2008 Wiley-VCH Verlag GmbH & Co. KGaA, Weinheim
www.chemeurj.org
8637

Z.-J. Yao et al.
+Na]; HRMS (MALDI): m/z: calcd for C38H70N2O8Na: 705.5024; found:
705.5035.
6.8 Hz), 7.01 (1H, s), 7.24 ppm (2H, brs); 13C NMR (125 MHz, CDCl3/
MeOD): d=175.6, 174.1, 149.1, 134.2, 77.5, 71.9, 34.5, 31.8, 29.5, 29.4,
29.3, 29.2, 29.1, 29.0, 27.3, 25.1, 25.0, 22.5, 19.0, 13.9 ppm; MS (ESI): m/z:
617 [M++Na]; HRMS (MALDI): m/z: calcd for C34H63N2O6Na:
595.4680; found: 595.4678.
Data for 61: Yellowish oil; yield: 75%; [a]2D5 =À53.8 (c=1.60 in CHCl3);
IR (film): n˜ =2925, 2854, 1756, 1655, 1466, 1406, 1318, 1151, 1105, 1039,
919, 722 cmÀ1 1H NMR (500 MHz, CDCl3): d=0.84 (3H, t, J=7.1 Hz),
;
1.22 (34H, m), 1.37 (3H, d, J=6.8 Hz), 1.52 (6H, m), 2.23 (2H, t, J=
7.0 Hz), 2.96 (1H, s), 3.07 (5H, s), 3.33 (6H, s), 3.31 (1H, m), 3.48 (2H,
m), 3.69 (1H, m), 4.32 (2H, m), 4.59 (4H, m), 4.95 (1H, dq, J=1.6,
6.8 Hz), 6.96 ppm (1H, d, J=1.5 Hz); 13C NMR (125 MHz, CDCl3): d=
173.7, 172.2, 148.7, 134.2, 95.5, 73.8, 55.87, 55.80, 55.7, 45.3, 35.2, 32.3,
31.8, 29.5, 29.4, 29.2, 29.1, 27.3, 25.6, 25.5, 25.1, 22.5, 19.1, 14.0 ppm; MS
(ESI): m/z: 719 [M++Na]; HRMS (MALDI): m/z: calcd for
C39H72N2O8Na: 719.5180; found: 719.5162.
Data for 67: White wax; yield: 57%; [a]2D5 =+10.3 (c=0.90, CHCl3); IR
(film): n˜ =3412, 2924, 2852, 2457, 1755, 1642, 1466, 1387, 1318, 1286,
1199, 1080, 1027, 722 cmÀ1 1H NMR (500 MHz, CDCl3): d=0.88 (3H, t,
;
J=7.0 Hz), 1.26 (36H, m), 1.40 (3H, d, J=6.8 Hz), 1.54 (4H, m), 2.26
(2H, t, J=8.0 Hz), 3.01 (6H, m),3.56 (4H+2OH, m), 4.30 (2H, m), 4.99
(1H, dq, J=1.5, 6.8 Hz), 7.01 ppm (1H, d, J=1.2 Hz); 13C NMR
(125 MHz, CDCl3): d=175.2, 173.8, 148.8, 134.3, 77.3, 68.1, 45.3, 34.9,
34.7, 31.8, 29.6, 29.5, 29.4, 29.3, 29.1, 27.4, 25.3, 25.1, 25.0, 22.6, 19.2,
14.0 ppm; MS (ESI): m/z: 631 [M++Na]; HRMS (MALDI): m/z: calcd
for C35H64N2O6Na: 631.4656; found: 631.4685.
Data for 62: Yellowish oil; yield: 73%; [a]2D5 =À28.4 (c=3.32 in CHCl3);
IR (film): n˜ =2925, 2854, 1756, 1651, 1463, 1371, 1318, 1216, 1153, 1105,
1041, 920, 722 cmÀ1
;
1H NMR (500 MHz, CDCl3): d=0.81 (3H, t, J=
Data for 68: White wax; yield: 70%; [a]2D5 =À2.43 (c=0.90 in CHCl3);
7.0 Hz), 1.19 (32H, m), 1.33 (3H, d, J=6.8 Hz), 1.46 (4H, m), 1.64 (4H,
m), 2.19 (2H, t, J=7.9 Hz), 3.30 (6H, s), 3.58 (8H, m), 4.30 (2H, s), 4.57
(4H, s), 4.92 (1H, dq, J=1.4, 6.8 Hz), 6.92 ppm (1H, d, J=1.4 Hz);
13C NMR (125 MHz, CDCl3): d=173.7, 170.7, 148.7, 134.2, 95.7, 75.3,
55.9, 45.3, 45.0, 42.2, 32.7, 31.7, 29.5, 29.4, 29.3, 29.2, 29.1, 27.3, 25.5, 25.0,
22.5, 19.1, 14.0 ppm; MS (ESI): m/z: 717 [M++Na]; HRMS (MALDI):
m/z: calcd for C39H70N2O8Na: 717.5024; found: 717.5034.
IR (film): n˜ =3419, 2924, 2853, 1754, 1641, 1467, 1395, 1319, 1283, 1255,
1082, 1022, 721 cmÀ1 1H NMR (500 MHz, CDCl3): d=0.88 (3H, t, J=
;
7.0 Hz), 1.29 (32H, m), 1.34 (3H, d, J=6.8 Hz), 1.49 (5H, m), 1.59 (3H,
m), 2.26 (2H, t, J=8.0 Hz), 3.43 (5H+2OH, m), 3.70 (2H, m), 3.83 (1H,
m), 4.36 (2H, m), 4.99 (1H, dq, J=1.4, 6.8 Hz), 6.98 ppm (1H, d, J=
1.2 Hz); 13C NMR (125 MHz, CDCl3): d=173.8, 148.8, 134.3, 77.3, 68.0,
44.6, 42.2, 35.1, 31.8, 29.6, 29.5, 29.4, 29.3, 29.2, 29.1, 27.4, 25.1, 24.9, 22.6,
19.2, 14.0 ppm; MS (ESI): m/z: 629 [M++Na]; HRMS (MALDI): m/z:
calcd for C35H62N2O6Na: 629.4500; found: 629.4513.
Data for 63: White solid; yield: 73%; [a]2D5 =À62.0 (c=3.35 in CHCl3);
IR (film): n˜ =3269, 3086, 2923, 2853, 1757, 1648, 1552, 1467, 1319, 1219,
1107, 1041, 919, 721 cmÀ1
;
1H NMR (500 MHz, CDCl3): d=0.80 (3H, t,
Data for 69: White wax; yield: 70%; [a]2D5 =À61.2 (c=1.25 in CHCl3);
J=7.0 Hz), 1.18 (38H, m), 1.33 (3H, d, J=6.8 Hz), 1.47 (2H, m), 1.64
(6H, m), 1.95 (2H, d, J=12.1 Hz), 2.19 (2H, t, J=7.9 Hz), 3.28 (6H, s),
3.63 (2H, m), 3.88 (2H, m), 4.44 (2H, d, J=6.8 Hz), 4.55 (2H, d, J=
6.8 Hz), 4.92 (1H, dq, J=1.5, 6.8 Hz), 6.68 (2H, brs), 6.91 ppm (1H, d,
J=1.4 Hz); 13C NMR (125 MHz, CDCl3): d=173.7, 172.6, 148.7, 134.2,
96.0, 77.4, 55.9, 52.69, 52.64, 32.9, 32.4, 31.8, 29.4, 29.3, 29.2, 29.1, 27.3,
25.1, 24.6, 24.5, 22.5, 19.1, 14.0 ppm; MS (ESI): m/z: 745 [M++Na];
HRMS (MALDI): m/z: calcd for C41H74N2O8Na: 745.5337; found:
745.5342.
IR (film): n˜ =3281, 3090, 2920, 2851, 1753, 1649, 1618, 1552, 1467, 1320,
1202, 1147, 1115, 1082, 1027, 950, 856, 721 cmÀ1 1H NMR (500 MHz,
;
CDCl3): d=0.88 (3H, t, J=7.1 Hz), 1.29 (38H, m), 1.40 (3H, d, J=
6.8 Hz), 1.55 (4H, m), 1.72 (4H, m), 1.94 (2H, m), 2.26 (2H, t, J=
8.0 Hz), 3.72 (2H, m), 3.98 (2H, m), 4.99 (1H, dq, J=1.6, 6.8 Hz), 6.98
(1H, d, J=1.4 Hz), 7.02 ppm (2H, m); 13C NMR (125 MHz, CDCl3): d=
175.8, 173.8, 148.8, 134.3, 77.3, 72.2, 53.2, 34.4, 32.0, 31.8, 29.6, 29.5, 29.4,
29.3, 29.2, 29.1, 27.4, 25.2, 25.1, 24.7, 22.6, 19.1, 14.0 ppm; MS (ESI): m/z:
657 [M++Na]; HRMS (MALDI): m/z: calcd for C37H66N2O6Na:
657.4813; found: 657.4814.
Data for 64: White solid; yield: 75%; [a]2D5 =À22.7 (c=3.45 in CHCl3);
IR (film): n˜ =3278, 3075, 2924, 2854, 1758, 1643, 1526, 1467, 1319, 1288,
Data for 70: White wax; yield: 70%; [a]2D5 =+20.5 (c=0.95 in CHCl3);
1154, 1103, 1032, 920, 721 cmÀ1 1H NMR (500 MHz, CDCl3): d=0.80
;
IR (film): n˜ =3340, 3260, 2919, 2451, 1752, 1651, 1617, 1539, 1467, 1320,
(3H, t, J=7.0 Hz), 1.18 (38H, m), 1.33 (3H, d, J=6.8 Hz), 1.49 (4H, m),
1.70 (4H, m), 2.03 (2H, d, J=11.8 Hz), 2.19 (2H, t, J=7.9 Hz), 3.32 (6H,
s), 3.59 (2H, m), 3.88 (2H, m), 4.56 (4H, s), 4.92 (1H, dq, J=1.6,
6.8 Hz), 6.77 (2H, brs), 6.91 ppm (1H, d, J=1.5 Hz); 13C NMR
(125 MHz, CDCl3): d=173.7, 173.2, 148.7, 134.2, 96.3, 78.2, 56.1, 52.9,
52.8, 33.4, 32.3, 31.8, 29.6, 29.5, 29.4, 29.3, 29.2, 29.1, 27.3, 25.5, 25.1, 24.5,
22.6, 19.1, 14.0 ppm; MS (ESI): m/z: 745 [M++Na]; HRMS (MALDI):
m/z: calcd for C41H74N2O8Na: 745.5337; found: 745.5333.
1202, 1114, 1082, 1027, 721 cmÀ1 1H NMR (500 MHz, CDCl3): d=0.87
;
(3H, t, J=7.1 Hz), 1.25 (38H, m), 1.40 (3H, d, J=6.8 Hz), 1.54 (4H, m),
1.77 (4H, m), 2.01 (2H, m), 2.26 (2H, t, J=8.0 Hz), 3.69 (2H, m), 4.00
(2H, m), 4.99 (1H, dq, J=1.5, 6.8 Hz), 6.88 (2H, brs), 6.98 ppm (1H, d,
J=1.4 Hz); 13C NMR (125 MHz, CDCl3): d=175.0, 173.9, 148.98, 134.3,
77.4, 72.0, 53.3, 35.0, 31.90, 29.6, 29.5, 29.4, 29.3, 29.2, 29.1, 27.4, 25.1,
24.6, 22.6, 19.1, 14.0 ppm; MS (ESI): m/z: 657 [M++Na]; HRMS
(MALDI): m/z: calcd for C37H66N2O6Na: 657.4813; found: 657.4822.
General procedure for 65–70:
A
solution of compounds 59–64
WST cell growth assay: The anticancer activities of newly synthesized
AA005 analogues were determined using the MDA-MB-468 and and
MDA-MB-435 human breast cancer cell lines. To test their selectivity,
normal mammary epithelial cells (HMEC) were used. Cells were seeded
in 96-well flat bottom cell culture plates at a density of 3000–4000 cells
per well with different concentrations of the tested compounds and incu-
bated for four days. Cell growth inhibition was determined using the
(0.10 mmol) in CH2Cl2 (2 mL) was treated with TFA (1 mL) at 08C.
After stirring at RT for 1 h, the solvent was removed under reduced pres-
sure. The residue was purified by column chromatography with CH2Cl2/
MeOH 20:1 to 10:1 to give compounds 65–70, respectively.
Data for 65: White wax; yield: 70%; [a]2D5 =À21.2 (c=0.42 in CHCl3);
IR (film): n˜ =3404, 3298, 2918, 2849, 2530, 2457, 1747, 1618, 1482, 1468,
1429, 1378, 1319, 1260, 1104, 1081, 1027, 951, 721 cmÀ1
;
1H NMR
WST-8
(2-(2-methoxy-4-nitrophenyl)-3-(4-nitrophenyl)-5-(2,4-disulfo-
phenyl)-2H-tetrazolium monosodium salt; Dojindo Molecular Technolo-
gies Inc., Gaithersburg, Maryland). WST-8 was added at a final concen-
tration of 10% to each well, and then the plates were incubated at 378C
for 2–3 h. The absorbance of the samples was measured at 450 nm using
a Molecular Device Reader. Concentration of the compounds that inhib-
ited cell growth by 50% (IC50) was calculated by comparing absorbance
in the untreated cells (DMSO control) and the cells treated with the
tested compounds.
(500 MHz, CDCl3/MeOD): d=0.88 (3H, t, J=7.1 Hz), 1.26 (34H, m),
1.41 (3H, d, J=6.8 Hz), 1.55 (4H, m), 1.78 (2H, m), 2.26 (2H, t, J=
8.0 Hz), 3.30 (2H, m), 3.46 (2H, m), 3.98 (2H, m), 5.00 (1H, dq, J=1.2,
6.8 Hz), 7.05 ppm (1H, J=1.5 Hz); 13C NMR (125 MHz, CDCl3/MeOD):
d=174.2, 172.3, 147.3, 131.8, 75.6, 69.5, 51.1, 36.6, 32.1, 29.6, 27.3, 27.2,
27.1, 27.0, 26.8, 25.1, 22.9, 20.3, 16.7 ppm; MS (ESI): m/z: 603 [M++Na];
HRMS (MALDI): m/z: calcd for C33H60N2O6Na: 603.4343; found:
603.4373.
Data for 66: White wax; yield: 65%; [a]2D5 =À11.0 (c=0.60 in CHCl3);
IR (film): n˜ =3401, 3299, 2919, 2848, 2523, 2456, 1748, 1614, 1538, 1484,
1468, 1437, 1376, 1317, 1080, 1027, 934, 721 cmÀ1 1H NMR (500 MHz,
;
CDCl3/MeOD): d=0.88 (3H, t, J=7.0 Hz), 1.26 (34H, m), 1.40 (3H, d,
J=6.8 Hz), 1.56 (4H, m), 1.72 (2H, m), 1.79 (2H, m), 2.26 (2H, t, J=
7.8 Hz), 3.21 (2H, m), 3.37 (2H, m), 4.02 (2H, m), 5.00 (1H, q, J=
Acknowledgements
This project is financially co-supported by the National Natural Science
Foundation (20425205, 20621062), Chinese Academy of Sciences
8638
www.chemeurj.org
ꢀ 2008 Wiley-VCH Verlag GmbH & Co. KGaA, Weinheim
Chem. Eur. J. 2008, 14, 8632 – 8639

Anticancer Agent AA005
FULL PAPER
(KGCX2-SW-213, KJCX2-YW-H08), and the Shanghai Municipal Com-
mittee of Science and Technology (06QH14016, 07DZ22001). Prof. S.
Wang and Dr. S. Qiu at the Comprehensive Cancer Center of the Univer-
sity of Michigan are greatly appreciated for their help in screening the in-
hibitory activities of samples.
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Received: June 28, 2008
Published online: August 7, 2008
Chem. Eur. J. 2008, 14, 8632 – 8639
ꢀ 2008 Wiley-VCH Verlag GmbH & Co. KGaA, Weinheim
www.chemeurj.org
8639