Solid-Phase Combinatorial Synthesis and Biological Evaluation of Destruxin e Analogues

Article abstract of DOI:10.1002/chem.201502970

The solid-phase combinatorial synthesis of cyclodepsipeptide destruxin E has been demonstrated. The combinatorial synthesis of cyclization precursors 8 was achieved by using a split and pool method on SynPhase Lanterns. The products were successfully macrolactonized in parallel in the solution phase by using 2-methyl-6-nitrobenzoic anhydride and 4-(dimethylamino)pyridine N-oxide to afford macrolactones 9, and the subsequent formation of an epoxide in the side chain gave 18 member destruxin E analogues 6. Biological evaluation of analogues 6 indicated that the N-MeAla residue was crucial to the induction of morphological changes in osteoclast-like multinuclear cells (OCLs). Based on structure-activity relationships, azido-containing analogues 15 were then designed for use as a molecular probe. The synthesis and biological evaluation of analogues 15 revealed that 15 b, in which the Ile residue was replaced with a Lys(N₃) residue, induced morphological changes in OCLs at a sufficient concentration, and modification around the Ile residue would be tolerated for attachment of a chemical tag toward the target identification of destruxin E (1). Forced to change: The 18 member analogues of destruxin E (see scheme) were successfully synthesized. Biological evaluation of these analogues indicated that the N-MeAla residue was crucial for inducing morphological changes in osteoclast-like multinuclear cells (OCLs). Molecular probes were then prepared by replacing the Ile residue with a Lys(N₃) residue for target identification in OCLs.

Full text of DOI:10.1002/chem.201502970

DOI: 10.1002/chem.201502970
Full Paper
&
Structure–Activity Relationships
Solid-Phase Combinatorial Synthesis and Biological Evaluation of
Destruxin E Analogues
Masahito Yoshida,[a] Yoshitaka Ishida,[a] Kenta Adachi,[a] Hayato Murase,[b]
Hiroshi Nakagawa,[b] and Takayuki Doi*[a]
Abstract: The solid-phase combinatorial synthesis of cyclo-
depsipeptide destruxin E has been demonstrated. The com-
binatorial synthesis of cyclization precursors 8 was achieved
by using a split and pool method on SynPhase Lanterns. The
products were successfully macrolactonized in parallel in the
solution phase by using 2-methyl-6-nitrobenzoic anhydride
and 4-(dimethylamino)pyridine N-oxide to afford macrolac-
tones 9, and the subsequent formation of an epoxide in the
side chain gave 18 member destruxin E analogues 6. Biologi-
cal evaluation of analogues 6 indicated that the N-MeAla
residue was crucial to the induction of morphological
changes in osteoclast-like multinuclear cells (OCLs). Based on
structure–activity relationships, azido-containing analogues
15 were then designed for use as a molecular probe. The
synthesis and biological evaluation of analogues 15 revealed
that 15b, in which the Ile residue was replaced with
a Lys(N3) residue, induced morphological changes in OCLs at
a sufficient concentration, and modification around the Ile
residue would be tolerated for attachment of a chemical tag
toward the target identification of destruxin E (1).
enzyme (BACE) or presenilin (PS)/g-secretase activity.[10] In par-
ticular, compound 1 is a more potent V-ATPase inhibitor than
any other member of the destruxin family[11] and could there-
fore be a promising candidate drug for use in cancer therapies
because V-ATPase plays an important role in allowing cancer
cells to behave invasively.[12] Intriguingly, it has recently been
reported that 1 induces morphological changes in osteoclast-
like multinuclear cells (OCLs) reversibly at a lower concentra-
tion than that required to affect V-ATPase activity in the OCLs,
resulting in inhibition of bone resorption without causing cell
death.[13] Most anti-resorption agents induce cell death at the
same time as inhibiting bone resorption; therefore 1 can be
considered as a new type of anti-resorption agent for use in
osteoporosis therapies. We wished to synthesize 1 because of
its structural features and unique biological activity, and we re-
cently achieved the first total synthesis of 1 and fully deter-
mined its structure. In particular, we found that the potency of
the V-ATPase inhibitory activity of 1 depends on the stereo-
chemistry of the epoxide moiety.[14] We therefore wished to
elucidate the mode of action of 1 in OCLs. Studying the
structure–activity relationships of the analogues of 1 would be
worthwhile because the information provided could allow
novel drug candidates for osteoporosis therapies to be devel-
oped.[15] Herein, we describe the combinatorial synthesis of an-
alogues of 1 and present the results of biological evaluations
of the analogues in terms of the morphological changes the
analogues bring about in OCLs.
Introduction
Cyclic peptides are novel compounds for use in the develop-
ment of drugs because they more easily permeate cells and
are more bioavailable than linear peptides.[1] Peptides such as
FK228,[2] cyclosporine A,[3] and daptomycin[4] that can be used
as drugs, not only have cyclic structures, but contain non-pro-
teinogenic amino acids, such as N-methylated or d-amino
acids. These features can give a drug unique pharmacological
properties.[5] This makes cyclic peptides containing non-protei-
nogenic amino acids attractive candidates for both the lead
discovery and lead optimization phases of drug development.
Destruxin E (1) was isolated from Metarhidium anisopliae by
Pa¨ıs et al. in 1981.[6a] Compound 1 has a 19-membered cyclo-
depsipeptide structure that consists of five proteinogenic and
non-proteinogenic amino acids (l-proline, l-isoleucine, N-Me-l-
valine, N-Me-l-alanine, and b-alanine) and an epoxide-contain-
ing derivative of a hydroxy acid.[6] It has recently been found
that 1 has several unique biological activities,[7] including anti-
proliferative activity against P388 leukemic cells,[8] the inhibi-
tion of the accumulation of lipid droplets in J774 macro-
phages,[9] and the ability to decrease the amount of amyloid-
b (Ab) generated without affecting the b-amyloid-cleaving
[a] Dr. M. Yoshida, Y. Ishida, K. Adachi, Prof.Dr. T. Doi
Graduate School of Pharmaceutical Sciences, Tohoku University
6-3 Aza-Aoba, Aramaki, Sendai 980-8578 (Japan)
E-mail: doi_taka@mail.pharm.tohoku.ac.jp
[b] H. Murase, Dr. H. Nakagawa
Department of Applied Biological Chemistry, Chubu University
1200 Matsumoto-cho, Kasugai, Aichi 487-8501 (Japan)
Supporting information for this article is available on the WWW under
http://dx.doi.org/10.1002/chem.201502970.
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chain was required to allow the molecule to affect the
osteoclast morphology at low concentrations.
Results and Discussion
Using the structural information presented above, we de-
signed destruxin E analogues composed of amino acid residues
present in destruxins and pseudodestruxins. The synthetic
strategy used to prepare the destruxin analogues 6 is shown
in Figure 3. Cyclization precursors 8 were synthesized in the
solid phase by using a split and pool method similar to the
method we previously used for the total synthesis of 1.[14] Mac-
rolactones 9 were formed through macrolactonization mediat-
ed by MNBA[17] in the solution phase after the reactants had
been cleaved from the polymer support. The formation of ep-
oxide moieties in parallel then gave the 18 member destruxin
analogues 6. Each of the analogues consisted of the amino
acids A (three), B (three), and C (two), as shown in Figure 3.
In brief, cyclization precursors 8 were synthesized in the
solid-phase by using the split and pool method[18] outlined in
Scheme 1. The 18 member b-Ala-OH-attached SynPhase Lan-
terns 10,[14,19] which were visualized by using colored stems
with cogs, were divided into three portions, and the first acyla-
tion was performed on each portion with a different amino
acid, A1 (N-Me-Ala), A2 (N-Me-Val), or A3 (N-Me-Leu), by using
DIC/HOBt (Scheme 1, step 1). The lanterns were then mixed
and washed with organic solvents to remove excess reagents.
The Fmoc group was removed with 20% piperidine in DMF,
and this left the polymer-supported amines 11, which were
split into three portions. The second coupling reaction was
performed on each portion with a different amino acid,
B1 (N-Me-Val), B2 (N-Me-Ile), or B3 (N-Me-Phe), by using
PyBroP[20]/DIEA. The lanterns were mixed and washed, and the
Fmoc group was removed to afford the polymer-supported
amines 12. Acylation of 12 to give tetrapeptide 13 was difficult
to achieve because of the presence of hindered N-Me-amines,
as we have previously reported;[14] therefore, amines 12 had to
be double coupled with Fmoc-Ile-OH C1 and Fmoc-Phe-OH C2
to give the tetrapeptides 13. Once the Fmoc group in 13 had
been removed, all of the polymer supports were acylated with
acid 7[21] to give hexapeptides 14. The TBS group was removed
with TBAF and then the products were cleaved in parallel from
the lanterns (with 30% HFIP/CH2Cl2 at room temperature for
1 h)[22] to give the 18 member cyclization precursors 8.[23]
The 18 member cyclization precursors 8 were macrolacton-
ized and then epoxide moieties were formed to produce des-
truxin E analogues 6, as shown in Scheme 2. Compound 8 was
subsequently macrolactonized with MNBA and DMAPO[17] to
provide the desired macrolactones 9 in 7–49% yields. Epoxide
moieties in the side chains were formed through three steps,
which have been described previously,[14] to give the 18
member destruxin E analogues 6 in moderate yields.
Synthesis and biological evaluation of destruxin E analogues
6
It is already known that
1 and destruxin B (2) induce
morphological changes in OCLs (Figure 1).[13] We assumed that
the biological activities of destruxins would depend on the
macrocyclic structures of the molecules. We also assumed that
similar structures, such as pseudodestruxin A (3) and pseudo-
destruxin B (4),[16] would also act as negative regulators of the
morphologies of osteoclasts (Figure 1).
We concluded that an epoxide moiety in the side chain
would be required to give a potent biological activity to the
molecule because 1 strongly affected the osteoclast morpho-
logy, whereas 2, which contained an isobutyl side chain, would
be less affected.[13] It seems, therefore, that the epoxide moiety
was fixed in the analogues presented. We first evaluated the
effect of the stereochemistry of the epoxide on osteoclast
morphology (Figure 2). Intriguingly, we found that 1, which
contained an (S)-epoxide moiety in the side chain, strongly
induced morphological changes in OCLs (0.04 mm), whereas
epi-destruxin E (5), previously reported,[14] which contains an
(R)-epoxide, affected the OCLs at a higher concentration
(5 mm). We concluded that an (S)-epoxide moiety in the side
Figure 1. Structures of destruxin E (1), destruxin B (2), pseudodestruxin A (3),
and pseudodestruxin B (4).
We evaluated the effects of destruxin E analogues 6 on the
morphologies of polarized OCLs, and the results are summar-
ized in Table 1. Among the analogues 6a–i, which contain an
isoleucine moiety in the R3 group, only analogues 6a–c, which
contain an N-Me-alanine residue in the R1 group, induced the
desired morphological changes without cell death (Table 1, en-
tries 1–3). Substitution of the R2 group with a more bulky or
hydrophobic group did not strongly negatively affect the bio-
Figure 2. Minimum concentration of 1 and epi-destruxin E (5) for morpho-
logical changes in OCLs.
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might allow the desired biologi-
cal activity to be retained, even
though introducing
a phenyl-
methyl group into the R3 group
decreased the biological activity.
Design, synthesis, and biologi-
cal evaluation of a molecular
probe 15 based on 1
Having recorded the structure–
activity relationships for mor-
phological changes in OCLs, we
next attempted the synthesis of
a molecular probe based on the
structure of 1. According to the
structure–activity relationships,
modification of the side chain
on the N-Me-Val or Ile residues
would be tolerant for inducing
morphological changes; thus we
designed molecular probes 15a
and 15b, which contained an
alkyl azido moiety to connect
a chemical tag, such as biotin or
FLAG tags, by copper-catalyzed
[3+2] cycloaddition.[24] In addi-
tion, we also planned to prepare
15c as a negative control be-
cause replacement of the Ala
residue diminished the desired
biological activity (Figure 4).
The synthesis of 15 was then
performed in the same manner
as that outlined above to pre-
pare cyclization precursors 16a–
c (Scheme 3), which contained
N-Me-Lys(N3)[25] or Lys(N3)[26], fol-
Figure 3. Synthetic strategy for producing the 18 member destruxin E analogues 6 and compositions of the
analogues in terms of amino acids A, B, and C. Fmoc=9-fluorenylmethoxycarbonyl, TBS=tert-butyldimethylsilyl,
SPPS=solid-phase peptide synthesis, MNBA=2-methyl-6-nitrobenzoic anhydride, DMAPO=4-(dimethylamino)-
pyridine N-oxide, TsCl=4-toluenesulfonyl chloride.
logical activity, although a slightly higher concentration was re-
quired to induce the desired morphological changes in the
OCLs. Analogues 6d–i were not effective at the concentrations
indicated in Table 1, but they were cytotoxic and induced cell
death (Table 1, entries 4–9). Therefore, we conclude that replac-
ing the R1 group with a bulky or hydrophobic moiety removed
the functionality we desire. We also conclude that the N-Me-
alanine residue plays a crucial role in inducing morphological
changes in OCLs. However, the desired biological activities
were lower for analogues 6j–r, in which R3 was a phenylmethyl
group, than those for analogues 6a–i (Table 1, entries 10–18).
Among analogues 6j–r, only 6j and 6k exhibited the desired
biological activity (Table 1, entries 10 and 11), and no effects
on the osteoclasts were observed for analogues 6l–r, which
exhibited cytotoxicity over the concentrations indicated in
Table 1, entries 12–18. Interestingly, analogue 6k, in which the
R2 was a bulky sec-butyl group, induced the desired morpho-
logical changes at a lower concentration than that of analogue
6j, which indicated that a bulky or hydrophobic R2 group
lowed by macrolactonization with MNBA/DMAPO to produce
macrolactones 17a–c. Finally, the formation of the epoxide in
the side chain gave derivatives 15a–c in moderate yields.
With the analogues 15a–c in hand, we evaluated the biolog-
ical activity and the results are summarized in Table 2. As ex-
pected, compound 15c as a negative control did not exhibit
the desired biological activity and cytotoxicity was observed at
a concentration of higher than 10 mm. Intriguingly, analogue
15b induced morphological changes in OCLs at 2.5 mm, where-
as analogue 15a required a higher concentration (10 mm) to
exhibit the activity, which indicated that modification around
the Ile residue would be tolerant for attaching a chemical tag,
and analogue 15b could be a good candidate as a molecular
probe toward target identification of 1.
Conclusion
We performed the combinatorial synthesis of 18 member des-
truxin E analogues 6 and evaluated the biological activities of
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phase by using a split and pool
method. The macrolactonization
of 8 with MNBA and DMAPO,
followed by the formation of ep-
oxide moieties in the side chains
in parallel, gave the 18 member
destruxin analogues 6 in moder-
ate yields. Biological evaluations
of the analogues showed that
modifying the N-Me-valine resi-
due allowed the molecule to
induce morphological changes,
but the N-Me-alanine residue
was essential for an analogue to
have the desired biological activ-
ity. This is the first time that the
relationship between the struc-
tures of destruxins and morpho-
logical changes induced has
been elucidated. In addition,
based on the above structure–
activity study, analogues 15
were designed as molecular
probes, and synthesized in an
analogous manner to that for
the synthesis of 6. Biological
evaluation of analogues 15a–
c indicated that 15b could be
a good candidate as a molecular
probe toward target identifica-
tion of 1. The results obtained
Scheme 1. Solid-phase synthesis of cyclization precursors 8 by using a split and pool method. a) Fmoc-AA-OH (A1,
A2, and A3) (0.1m), diisopropylcarbodiimide (DIC; 0.1m), 1-hydroxybenzotriazole (HOBt; 0.15m), DMF, RT, 12 h;
b) 20% piperidine/DMF, RT, 1 h; c) Fmoc-AA-OH (B1, B2, and B3) (0.1m), bromotripyrrolidinophosphonium hexa-
fluorophosphate (PyBroP; 0.1m), ethyldiisopropylamine (DIEA; 0.15m), DMF, RT, 12 h; d) Fmoc-AA-OH (C1 and C2)
(0.1m), PyBroP (0.1m), DIEA (0.15m), DMF, RT, 12 h, repeated; e) 7 (0.1m), PyBroP (0.1m), DIEA (0.15m), DMF, RT,
12 h; f) tetrabutylammonium fluoride (TBAF), THF, 08C, 4 h; g) 30% hexafluoroisopropanol (HFIP)/CH2Cl2, RT, 1 h.
will allow novel lead structures for anti-resorption
agents based on cyclodepsipeptide scaffolds to be
developed.
Experimental Section
General
All commercially available reagents were used as re-
ceived. Dry THF and CH2Cl2 (Kanto Chemical Co.) were
obtained by passing commercially available predried,
oxygen-free formulations through activated alumina col-
umns. MeOH was distilled from iodide and magnesium
turnings. DMF was purchased from Wako Chemical Ind.,
Ltd. (for peptide synthesis, grade: 99.5%). Trityl alcohol
lantern was purchased from Mimotopes (surface: poly-
styrene, loading: 35 mmol). Fmoc-protected amino
acids (Fmoc-bAla-OH, Fmoc-MeAla-OH, Fmoc-Ala-
OH·H2O, Fmoc-MeVal-OH, Fmoc-MeLeu-OH, Fmoc-MeIle-
OH, Fmoc-MePhe-OH, Fmoc-Ile-OH, and Fmoc-Phe-OH)
were purchased from Watanabe Chemical Ind., Ltd. All
reactions in solution were monitored by TLC carried out
on 0.2 mm Merck silica gel plates (60F-254) and visual-
ized with UV light, anisaldehyde, or 10% phosphomolyb-
Scheme 2. Synthesis of destruxin analogues 6 from cyclization precursors 8. a) MNBA,
DMAPO, CH2Cl2, RT, 308C, 48 h; b) aqueous 1m HCl, dioxane·H2O (1:2), 08C, 1 h; c) TsCl,
NEt3, 4-dimethylaminopyridine (DMAP), CH2Cl2, RT, 12 h; d) K2CO3, iPrOH·(CH2Cl)2, 508C,
12 h.
dic acid in ethanol. Silica gel 60N (Kanto Chemical Co.
1
the analogues in terms of the effects they had on the mor-
100~210 mm) was used for column chromatography. H (400 MHz)
and 13C NMR spectra (100 MHz) were recorded on JEOL JNM-AL400
phologies of OCLs. The 18 member cyclization precursors 8
were efficiently synthesized with good purities in the solid
1
spectrometers. H (600 MHz) and 13C NMR spectra (150 MHz) were
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Table 1. Evaluation of the effects of destruxin E analogues 6 on osteo-
Table 2. Evaluation of the effects of 15 on osteoclast morphology.
clast morphology.
Entry
Analogue
Yield [%][a]
[mm][b]
Entry
Analogue
{A, B, C}
Yield [%][a]
[mm][b]
1
2
3
15a
15b
15c
8
9
6
10
2.5
(10)[c]
1
2
3
4
5
6
6a[c]
6b
6c
6d
6e
6 f
{1, 1, 1}
{1, 2, 1}
{1, 3, 1}
{2, 1, 1}
{2, 2, 1}
{2, 3, 1}
{3, 1, 1}
{3, 2, 1}
{3, 3, 1}
{1, 1, 2}
{1, 2, 2}
{1, 3, 2}
{2, 1, 2}
{2, 2, 2}
{2, 3, 2}
{3, 1, 2}
{3, 2, 2}
{3, 3, 2}
13
12
8
3
2
7
4
8
5
9
13
7
3
3
7
2
5
10
0.04
0.1
10
(5.0)
(10)
(2.5)
(6.3)
(2.5)
(0.06)
12.5
3.1
(25)
(12.5)
(25)
[a] Yield of product isolated, as calculated from b-Ala-OH-attached lantern
10. [b] Minimum concentration required to induce morphological
changes. [c] The value in parentheses is the maximum concentration for
cell survival.
7
8
9
6g
6h
6i
10
11
12
13
14
15
16
17
18
6j
6k
6l
6m
6n
6o
6p
6q
6r
recorded on JEOL JNM-ECA600 spectrometers in the indicated sol-
vent. Chemical shifts (d) are reported in ppm relative to the signal
for internal tetramethylsilane (TMS; d=0 ppm for H) for solutions
in CDCl3 or residual protonated solvent: CDCl3 (7.26 ppm for H or
d=77.0 ppm for 13C) or [D6]DMSO (d=2.49 ppm for H). Multiplici-
ties are reported as follows: s (singlet), d (doublet), t (triplet), q
(quartet), quin (quintet), m (multiplet) dd (doublet of doublet), dt
(doublet of triplet), dquin (doublet of quintet), ddd (doublet of
doublet of doublet), brt (broad triplet); coupling constants, J, are
given in Hz. IR spectra were recorded on a JASCO FT/IR-4100 spec-
trometer. Only the strongest and/or structurally important absorp-
tions are recorded in cmÀ1. Optical
1
1
(25)
1
(12.5)
(3.5)
(25)
[a] Yield of product isolated, as calculated from b-Ala-OH-attached lantern
10. [b] Minimum concentration required to induce morphological
changes. Values in parentheses are maximum concentrations for cell
survival. [c] Destruxin E (1).
rotations were measured on
a JASCO P-1000 polarimeter. Melt-
ing points were recorded on
a
Round Science RFS-10 instru-
ment and are uncorrected. Mass
spectra (FAB) and high-resolution
mass spectra (FAB) were measured
on JEOL JMS-DX303 and JMS-700
instruments. Preparative HPLC pu-
rification was performed on re-
versed-phase HPLC (Waters HPLC
system), and the purity was deter-
mined with peak area at l=
214 nm. The column used was an
X Bridge C18 5 mm, 10”150 mm
(Waters) column. The gradient
method was as follows: 10% B
(0.00–1.00 min) 10–90% B (1.00–
13.0 min), 90% B (13.0–18.0 min),
95–10% B (18.0–19.0 min), 10% B
Figure 4. Design of a molecular probe containing an alkyl azido moiety.
(19.0–22.0 min)
(A:
0.100%
HCOOH/H2O, B: 0.100% HCOOH/
MeOH).
General procedure I: Macrolac-
tonization of 8
DMAPO (2.00 equiv) and MNBA
(3.00 equiv) were added to a solu-
tion of crude cyclization precursor
8 (1 equiv) in dry CH2Cl2 (3.00 mm)
at room temperature under an
argon atmosphere. After being
stirred at 308C for 48 h, the reac-
tion mixture was poured into a sa-
turated aqueous solution of
NaHCO3 and the aqueous layer
was extracted with CHCl3. The or-
Scheme 3. Synthesis of molecular probes 15. a) i) Fmoc-Ala-OH (for 15a and 15b) or Fmoc-N-Me-Lys(N3)-OH (for
15c) (0.1m), DIC (0.1m), HOBt (0.15m), DMF, RT, 12 h; ii) 20% piperidine/DMF, RT, 1 h; b) i) Fmoc-Me-Val-OH (for
15b and 15c) or Fmoc-N-Me-Lys(N3)-OH (for 15a), PyBroP (0.1m), DIEA (0.15m), DMF, RT, 12 h; ii) 20% piperidine/
DMF, RT, 1 h; c) i) Fmoc-Ile-OH (for 15a and 15c) or Fmoc-Lys(N3)-OH (for 15b), PyBroP (0.1m), DIEA (0.15m), DMF,
RT, 12 h, repeated; ii) 20% piperidine/DMF, RT, 1 h; d) 7 (0.1m), PyBroP (0.1m), DIEA (0.15m), DMF, RT, 12 h;
e) i) TBAF, THF, 08C, 4 h; ii) 30% HFIP/CH2Cl2, RT, 1 h; f) MNBA, DMAPO, CH2Cl2, RT, 308C, 48 h; g) aqueous 1m HCl,
dioxane/H2O (1:2), 08C, 1 h; h) TsCl, NEt3, DMAP, CH2Cl2, RT, 12 h; i) K2CO3, iPrOH/(CH2Cl)2, 508C, 12 h.
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ganic layer was washed with brine, dried over MgSO4, and filtered.
The filtrate was concentrated in vacuo, and the resulting residue
was purified by column chromatography on silica gel (eluted with
CHCl3/MeOH=40:1) and preparative HPLC to afford macrolatone 9
as a colorless oil.
Compound 9d {2, 1, 1}: Yield: 15% from polymer-supported H-
bAla-OH 10 (10.6 mg, 15.6 mmol); [a]2D4 =À133 (c=0.230, CHCl3);
1H NMR (400 MHz, CDCl3): d=8.10 (d, J=10.0 Hz, 1H), 7.15 (d, J=
8.8 Hz, 1H), 5.24 (d, J=10.8 Hz, 1H), 5.10 (dd, J=6.0, 8.8 Hz, 1H),
4.63–4.70 (m, 2H), 4.37 (d, J=10.4 Hz, 1H), 4.05–4.13 (m, 2H),
3.97–4.04 (m, 1H), 3.95 (t, J=8.0 Hz, 1H), 3.75–3.83 (m, 1H), 3.62 (t,
J=6.8 Hz, 1H), 3.15 (s, 3H), 3.12 (brt, J=12.4 Hz, 1H), 2.94 (s, 3H),
2.67 (dd, J=5.6, 18.4 Hz, 1H), 2.53 (dd, J=10.0, 18.4 Hz, 1H), 2.40–
2.47 (m, 2H), 2.33–2.38 (m, 1H), 2.15–2.24 (m, 1H), 2.05–2.13 (m,
1H), 1.93–2.04 (m, 4H), 1.47–1.54 (m, 1H), 1.42 (s, 3H), 1.33 (s, 3H),
1.24–1.32 (m, 1H), 0.93 (d, J=6.8 Hz, 3H), 0.90 (d, J=6.8 Hz, 3H),
0.89 (d, J=6.8 Hz, 3H), 0.88 (t, J=7.2 Hz, 3H), 0.84 (d, J=7.2 Hz,
3H), 0.78 ppm (d, J=7.2 Hz, 3H); 13C NMR (100 MHz, CDCl3): d=
173.6, 173.2, 171.3, 170.6, 169.3, 168.5, 109.3, 71.5, 71.3, 69.2, 65.9,
60.8, 56.8, 53.7, 46.4, 37.3, 34.5, 34.1, 32.9, 30.5, 29.2, 29.1, 28.6,
27.2, 26.9, 25.7, 25.5, 24.0, 20.4, 19.8, 19.3, 18.7, 14.4, 11.0 ppm; IR
(CHCl3): n˜ =3386, 3276, 2966, 2933, 2879, 1727, 1672, 1626, 1519,
1446, 1414, 1380, 1224, 1181, 1059, 751 cmÀ1; HRMS (FAB): m/z
calcd for C34H58N5O9 [M+H]+: 680.4235; found: 680.4242.
Compound 9a {1, 1, 1}: Yield: 38% from polymer-supported H-
bAla-OH 10 (26.0 mg, 39.9 mmol); [a]2D4 =À216 (c=0.606, CHCl3);
1H NMR (400 MHz, CDCl3): d=8.19 (d, J=7.8 Hz, 1H), 7.14 (d, J=
9.3 Hz, 1H), 5.15 (q, J=6.8 Hz, 1H), 5.05 (dd, J=5.9, 8.0 Hz, 1H),
4.96 (d, J=11.0 Hz, 1H), 4.89 (dd, J=6.3, 9.0 Hz, 1H), 4.68 (d, J=
6.8 Hz, 1H), 4.00–4.11 (m, 3H), 3.90 (brt, J=7.6 Hz, 1H), 3.75–3.81
(m, 1H), 3.62 (t, J=6.3 Hz, 1H), 3.22 (s, 3H), 3.08 (brt, J=12.7 Hz,
1H), 2.72 (s, 3H), 2.47–2.67 (m, 3H), 2.27–2.37 (m, 1H), 1.88–2.19
(m, 5H), 1.42 (s, 3H), 1.32 (s, 3H), 1.30 (d, J=6.8 Hz, 3H), 1.25–1.42
(m, 2H), 0.93 (d, J=6.6 Hz, 3H), 0.89 (d, J=6.6 Hz, 3H), 0.87 (d, J=
6.8 Hz, 3H), 0.85 ppm (t, J=7.3 Hz, 3H); 13C NMR (100 MHz, CDCl3):
d=173.5, 173.4, 171.0, 170.8, 169.7, 168.9, 109.2, 71.6, 70.8, 69.1,
60.8, 58.0, 55.5, 53.4, 46.4, 37.5, 34.5, 34.4, 33.2, 30.8, 29.2, 28.1,
27.2, 26.8, 25.5, 24.3, 23.9, 20.0, 19.6, 15.4, 15.1, 11.4 ppm; IR (neat):
n˜ =2966, 1732, 1668, 1627, 1520, 1446, 1379, 1180 cmÀ1; HRMS
(FAB): m/z calcd for C32H54N5O9 [M+H]+: 652.3916; found:
652.3898.
Compound 9e {2, 2, 1}: Yield: 7.0% from polymer-supported H-
bAla-OH 10 (5.1 mg, 7.35 mmol); [a]2D4 =À160 (c=0.230, CHCl3);
1H NMR (400 MHz, CDCl3): d=8.11 (d, J=10.0 Hz, 1H), 7.15 (d, J=
8.8 Hz, 1H), 5.29 (d, J=10.8 Hz, 1H), 5.10 (dd, J=6.0, 8.4 Hz, 1H),
4.69 (d, J=6.4 Hz, 1H), 4.66 (t, J=8.8 Hz, 1H), 4.36 (d, J=10.4 Hz,
1H), 3.97–4.12 (m, 3H), 3.93 (t, J=8.4 Hz, 1H), 3.75–3.83 (m, 1H),
3.62 (t, J=6.4 Hz, 1H), 3.13 (s, 3H), 3.04–3.17 (m, 1H), 2.94 (s, 3H),
2.67 (dd, J=5.6, 18.0 Hz, 1H), 2.53 (dd, J=10.0, 18.0 Hz, 1H), 2.33–
2.47 (m, 2H), 2.15–2.24 (m, 3H), 2.06–2.15 (m, 2H), 1.93–2.05 (m,
2H), 1.46–1.54 (m, 1H), 1.42 (s, 3H), 1.33–1.41 (m, 1H), 1.33 (s, 3H),
1.24–1.32 (m, 1H), 0.96–1.07 (m, 1H), 0.94 (d, J=6.8 Hz, 3H), 0.89
(t, J=7.2 Hz, 3H), 0.87 (d, J=6.8 Hz, 3H), 0.86 (t, J=6.8 Hz, 3H),
0.86 (d, J=6.4 Hz, 3H), 0.78 ppm (d, J=6.8 Hz, 3H); 13C NMR
(100 MHz, CDCl3): d=173.7, 173.1, 171.4, 170.7, 169.4, 168.7, 109.3,
71.5, 71.3, 69.2, 65.9, 60.8, 55.5, 53.7, 46.5, 37.3, 34.5, 34.1, 33.4,
33.0, 30.6, 29.3, 29.2, 28.6, 26.9, 25.7, 25.5, 24.7, 24.0, 19.8, 19.3,
16.6, 14.4, 11.1, 11.0 ppm; IR (CHCl3): n˜ =3383, 3271, 2966, 2933,
2877, 1727, 1671, 1625, 1522, 1449, 1413, 1380, 1225, 1180, 1077,
1059, 848, 751 cmÀ1; HRMS (FAB): m/z calcd for C35H60N5O9 [M+H]+
: 694.4391; found: 694.4388.
Compound 9b {1, 2, 1}: Yield: 27% from polymer-supported H-
bAla-OH 10 (18.9 mg, 28.4 mmol); [a]2D4 =À187 (c=0.350, CHCl3);
1H NMR (400 MHz, CDCl3): d=8.21 (d, J=8.8 Hz, 1H), 7.14 (d, J=
8.8 Hz, 1H), 5.14 (q, J=6.8 Hz, 1H), 5.06 (t, J=6.8 Hz, 1H), 5.04 (d,
J=11.2 Hz, 1H), 4.89 (dd, J=6.8, 8.8 Hz, 1H), 4.68 (d, J=6.8 Hz,
1H), 4.00–4.12 (m, 3H), 3.91 (t, J=8.0 Hz, 1H), 3.75–3.82 (m, 1H),
3.62 (t, J=6.8 Hz, 1H), 3.20 (s, 3H), 3.08 (brt, J=13.2 Hz, 1H), 2.72
(s, 3H), 2.50–2.65 (m, 2H), 2.46 (d, J=6.0 Hz, 1H), 2.11–2.20 (m,
2H), 2.03–2.10 (m, 2H), 1.84–2.00 (m, 5H), 1.42 (s, 3H), 1.32 (s, 3H),
1.30 (d, J=6.8 Hz, 3H), 1.25–1.32 (m, 2H), 0.92 (d, J=5.6 Hz, 3H),
0.87 (t, J=6.8 Hz, 3H), 0.86 (d, J=6.8 Hz, 3H), 0.85 ppm (t, J=
6.8 Hz, 3H); 13C NMR (100 MHz, CDCl3): d=173.5, 173.4, 171.1,
170.9, 169.7, 168.9, 109.2, 71.6, 70.8, 69.1, 60.8, 56.7, 55.5, 53.6,
46.4, 37.5, 34.6, 34.5, 33.5, 33.2, 31.0, 29.3, 28.1, 26.9, 25.7, 25.5,
24.4, 23.9, 16.2, 15.4, 15.2, 11.4, 11.0 ppm; IR (neat): n˜ =3383, 3295,
2965, 2934, 2878, 1731, 1667, 1629, 1518, 1445, 1413, 1380, 1230,
1179, 1106, 1059 cmÀ1; HRMS (FAB): m/z calcd for C33H55N5O9 [M+
H]+: 666.4078; found: 666.4091.
Compound 9 f {2, 3, 1}: Yield: 26% from polymer-supported H-
bAla-OH 10 (19.9 mg, 27.3 mmol); [a]2D5 =À163 (c=0.650, CHCl3);
1H NMR (400 MHz, CDCl3): d=8.36 (d, J=9.2 Hz, 1H), 7.17–7.31 (m,
5H), 7.14 (d, J=9.2 Hz, 1H), 5.81 (dd, J=6.8, 8.4 Hz, 1H), 5.08 (dd,
J=6.0, 8.0 Hz, 1H), 4.84 (dd, J=6.4, 9.2 Hz, 1H), 4.72 (d, J=6.8 Hz,
1H), 4.35 (d, J=10.4 Hz, 1H), 4.03–4.14 (m, 3H), 3.90 (t, J=8.0 Hz,
1H), 3.76–3.84 (m, 1H), 3.63 (t, J=6.8 Hz, 1H), 3.21–3.27 (m, 2H),
3.20 (s, 3H), 3.08–3.19 (m, 1H), 2.80 (s, 3H), 2.55–2.67 (m, 2H), 2.46
(d, J=6.4 Hz, 1H), 2.23–2.31 (m, 1H), 2.08–2.21 (m, 2H), 2.01–2.08
(m, 1H), 1.91–1.99 (m, 2H), 1.83–1.90 (m, 1H), 1.43 (s, 3H), 1.36–1.47
(m, 1H), 1.33 (s, 3H), 1.20–1.32 (m, 1H), 0.89 (d, J=6.8 Hz, 3H), 0.84
(d, J=6.8 Hz, 3H), 0.84 (t, J=7.2 Hz, 3H), 0.49 ppm (d, J=6.8 Hz,
3H); 13C NMR (100 MHz, CDCl3): d=173.4, 172.8, 171.4, 170.5, 168.9,
168.6, 136.0, 128.9, 128.5, 127.0, 109.2, 71.5, 70.9, 69.1, 66.3, 60.7,
53.4, 51.3, 46.3, 37.6, 35.0, 34.5, 34.4, 32.8, 31.7, 29.3, 29.0, 27.1, 26.8,
25.5, 24.3, 23.8, 19.6, 18.4, 15.4, 11.3 ppm; IR (neat): n˜ =3386, 3299,
2964, 2934, 2876, 1731, 1671, 1629, 1519, 1499, 1446, 1412, 1381,
1370, 1337, 1222, 1181, 1103, 1059, 754, 701 cmÀ1; HRMS (FAB): m/z
calcd for C38H58N5O9 [M+H]+: 728.4235; found: 728.4213.
Compound 9c {1, 3, 1}: Yield: 26% from polymer-supported H-
bAla-OH 10 (19.1 mg, 27.3 mmol); [a]2D4 =À146 (c=0.560, CHCl3);
1H NMR (400 MHz, CDCl3): d=8.25 (d, J=7.2 Hz, 1H), 7.14–7.30 (m,
5H), 7.11 (d, J=9.2 Hz, 1H), 5.39 (dd, J=4.0, 12.0 Hz, 1H), 5.04 (dd,
J=6.0, 8.0 Hz, 1H), 4.98 (dd, J=6.4, 9.2 Hz, 1H), 4.70 (d, J=6.8 Hz,
1H), 4.66 (q, J=6.8 Hz, 1H), 4.02–4.12 (m, 2H), 3.88–3.98 (m, 2H),
3.75–3.84 (m, 1H), 3.62 (t, J=6.4 Hz, 1H), 3.35 (s, 3H), 3.22–3.38 (m,
1H), 3.05 (brt, J=13.2 Hz, 1H), 2.92 (dd, J=4.0, 12.0 Hz, 1H), 2.56
(s, 3H), 2.51–2.66 (m, 2H), 2.46 (d, J=6.8 Hz, 1H), 2.11–2.18 (m,
2H), 2.05–2.10 (m, 2H), 1.93–2.01 (m, 1H), 1.81–1.90 (m, 1H), 1.42
(s, 3H), 1.33 (s, 3H), 1.22–1.30 (m, 2H), 0.92 (d, J=6.8 Hz, 3H), 0.86
(t, J=6.8 Hz, 3H), 0.18 ppm (d, J=6.8 Hz, 3H); 13C NMR (100 MHz,
CDCl3): d=173.3, 172.6, 170.9, 170.7, 169.8, 168.9, 136.0, 129.5,
128.9, 127.3, 109.2, 71.6, 70.6, 69.1, 60.8, 55.6, 53.9, 53.2, 46.4, 37.7,
36.5, 34.5, 34.4, 33.2, 31.5, 29.5, 28.0, 26.8, 25.5, 24.3, 23.9, 15.6,
13.1, 11.3 ppm; IR (neat): n˜ =3383, 3305, 2965, 2935, 2878, 1731,
1664, 1636, 1518, 1493, 1448, 1412, 1379, 1337, 1228, 1191, 1109,
1060, 752, 702, 679 cmÀ1; HRMS (FAB): m/z calcd for C36H54N5O9
[M+H]+: 700.3922; found: 700.3911.
Compound 9g {3, 1, 1}: Yield: 23% from polymer-supported H-
bAla-OH 10 (16.9 mg, 24.4 mmol); [a]2D4 =À195 (c=0.845, CHCl3);
Chem. Eur. J. 2015, 21, 18417 – 18430
www.chemeurj.org
18422
ꢀ 2015 Wiley-VCH Verlag GmbH & Co. KGaA, Weinheim

Full Paper
1H NMR (400 MHz, CDCl3): d=8.32 (d, J=9.6 Hz, 1H), 7.17 (d, J=
9.2 Hz, 1H), 5.06 (dd, J=6.0, 8.0 Hz, 1H), 5.00 (d, J=10.8 Hz, 1H),
4.94 (dd, J=6.0, 9.2 Hz, 1H), 4.89 (dd, J=2.0, 11.6 Hz, 1H), 4.72 (d,
J=6.8 Hz, 1H), 4.03–4.16 (m, 3H), 3.89 (t, J=8.0 Hz, 1H), 3.76–3.84
(m, 1H), 3.63 (t, J=6.8 Hz, 1H), 3.22 (s, 3H), 3.09 (brt, J=12.0 Hz,
1H), 2.76 (s, 3H), 2.56–2.64 (m, 2H), 2.50 (d, J=6.4 Hz, 1H), 2.36–
2.41 (m, 1H), 2.29–2.36 (m, 1H), 2.09–2.21 (m, 2H), 2.01–2.08 (m,
1H), 1.93–2.00 (m, 2H), 1.85–1.90 (m, 1H), 1.47–1.58 (m, 1H), 1.42
(s, 3H), 1.33–1.42 (m, 1H), 1.33 (s, 3H), 1.25–1.32 (m, 1H), 0.99–1.05
(m, 1H), 0.97 (d, J=6.8 Hz, 3H), 0.93 (d, J=6.8 Hz, 6H), 0.90 (d, J=
6.4 Hz, 3H), 0.85 (d, J=6.8 Hz, 3H), 0.83 ppm (t, J=7.2 Hz, 3H);
13C NMR (100 MHz, CDCl3): d=173.4, 173.1, 170.7, 170.6, 168.8,
168.6, 109.2, 71.6, 70.8, 69.1, 60.7, 58.6, 57.7, 53.5, 46.3, 39.2, 37.5,
34.5, 34.4, 32.8, 30.8, 29.3, 28.7, 27.4, 26.8, 25.6, 25.5, 24.2, 23.8,
23.6, 22.5, 20.0, 19.7, 15.5, 11.4 ppm; IR (neat): n˜ =3387, 3299,
2962, 2936, 2875, 1732, 1673, 1634, 1516, 1446, 1414, 1380, 1370,
1343, 1218, 1181, 1061, 751 cmÀ1; HRMS (FAB): m/z calcd for
C35H60N5O9 [M+H]+: 694.4391; found: 694.4376.
Compound 9j {1, 1, 2}: Yield: 34% from polymer-supported H-
bAla-OH 10 (24.5 mg, 35.7 mmol); [a]2D9 =À132 (c=1.23, CHCl3);
1H NMR (400 MHz, CDCl3): d=7.84 (d, J=8.4 Hz, 1H), 7.34 (d, J=
9.2 Hz, 1H), 7.18–7.30 (m, 5H), 5.30 (q, J=6.8 Hz, 1H), 5.08–5.15
(m, 1H), 5.07 (t, J=6.8 Hz, 1H), 5.03 (d, J=11.2 Hz, 1H), 4.61 (d, J=
7.6 Hz, 1H), 4.03–4.15 (m, 3H), 3.58–3.70 (m, 3H), 3.23 (s, 3H),
3.12–3.19 (m, 1H), 3.01–3.07 (m, 1H), 2.81–2.86 (m, 1H), 2.81 (s,
3H), 2.63–2.69 (m, 2H), 2.28–2.40 (m, 1H), 2.16–2.21 (m, 1H), 2.10–
2.16 (m, 2H), 1.70–1.82 (m, 2H), 1.43 (s, 3H), 1.33 (d, J=6.8 Hz,
3H), 1.32 (s, 3H), 1.22–1.30 (m, 1H), 0.93 (d, J=6.8 Hz, 3H),
0.88 ppm (d, J=6.4 Hz, 3H); 13C NMR (100 MHz, CDCl3): d=173.6,
173.5, 170.7, 170.1, 169.7, 168.5, 136.5, 129.1, 128.4, 126.9, 109.1,
71.6, 70.9, 69.1, 60.3 58.0, 55.3, 51.1, 46.3, 37.5, 34.6, 34.3, 33.2,
30.3, 28.5, 28.1, 27.4, 26.8, 25.5, 23.2, 19.8, 19.3, 15.1 ppm; IR (neat):
n˜ =3385, 3307, 2980, 2936, 2879, 1728, 1668, 1634, 1531, 1446,
1414, 1380, 1338, 1228, 1182, 1061, 753, 701 cmÀ1; HRMS (FAB): m/
z calcd for C35H52N5O9 [M+H]+: 686.3765; found: 686.3765.
Compound 9k {1, 2, 2}: Yield: 33% from polymer-supported H-
bAla-OH 10 (24.2 mg, 34.7 mmol); [a]2D9 =À152 (c=1.08, CHCl3);
1H NMR (400 MHz, CDCl3): d=7.85 (d, J=8.4 Hz, 1H), 7.32 (d, J=
9.6 Hz, 1H), 7.19–7.30 (m, 5H), 5.29 (q, J=6.8 Hz, 1H), 5.10–5.15
(m, 1H), 5.10 (d, J=11.2 Hz, 1H), 5.07 (t, J=6.8 Hz, 1H), 4.61 (d, J=
7.6 Hz, 1H), 4.05–4.13 (m, 3H), 3.59–3.70 (m, 3H), 3.21 (s, 3H),
3.12–3.20 (m, 1H), 3.03 (dd, J=4.4, 13.6 Hz, 1H), 2.82 (dd, J=11.2,
13.6 Hz, 1H), 2.81 (s, 3H), 2.62–2.69 (m, 2H), 2.05–2.21 (m, 3H),
1.71–1.83 (m, 2H), 1.43–1.51 (m, 1H), 1.43 (s, 3H), 1.32 (d, J=
7.2 Hz, 3H), 1.31 (s, 3H), 1.17–1.28 (m, 1H), 0.95–1.05 (m, 1H), 0.92
(t, J=7.2 Hz, 3H), 0.84 ppm (d, J=6.8 Hz, 3H); 13C NMR (100 MHz,
CDCl3): d=173.6, 173.4, 170.7, 170.2, 169.7, 168.5, 136.6, 129.1,
128.5, 126.9, 109.1, 71.6, 70.9, 69.1, 60.3, 56.6, 55.3, 51.2, 46.3, 37.5,
34.6, 34.3, 33.6, 33.2, 30.4, 28.6, 28.1, 26.8, 25.5, 25.4, 23.2, 16.0,
15.1, 11.0 ppm; IR (neat): n˜ =3388, 3299, 2967, 2936, 2877, 1728,
1670, 1634, 1532, 1445, 1414, 1379, 1340, 1228, 1183, 1061, 753,
701, 665 cmÀ1; HRMS (FAB): m/z calcd for C36H54N5O9 [M+H]+:
700.3922; found: 700.3929.
Compound 9h {3, 2, 1}: Yield: 32% from polymer-supported H-
bAla-OH 10 (23.5 mg, 33.2 mmol); [a]2D5 =À162 (c=1.18, CHCl3);
1H NMR (400 MHz, CDCl3): d=8.34 (d, J=10.0 Hz, 1H), 7.16 (d, J=
9.2 Hz, 1H), 5.09 (d, J=10.8 Hz, 1H), 5.07 (dd, J=6.0, 8.0 Hz, 1H),
4.93 (dd, J=6.4, 9.2 Hz, 1H), 4.89 (d, J=12.0 Hz, 1H), 4.72 (d, J=
6.8 Hz, 1H), 4.03–4.17 (m, 3H), 3.89 (t, J=8.0 Hz, 1H), 3.76–3.84 (m,
1H), 3.62 (t, J=6.4 Hz, 1H), 3.20 (s, 3H), 3.09 (brt, J=12.0 Hz, 1H),
2.76 (s, 3H), 2.57–2.64 (m, 2H), 2.47 (d, J=6.0 Hz, 1H), 2.38 (dt,
J=4.0, 12.8 Hz, 1H), 2.11–2.22 (m, 3H), 2.04–2.11 (m, 2H), 1.92–2.00
(m, 1H), 1.83–1.91 (m, 1H), 1.47–1.58 (m, 1H), 1.42 (s, 3H), 1.38–
1.47 (m, 2H), 1.33 (s, 3H), 1.25–1.32 (m, 1H), 0.97–1.05 (m, 2H),
0.97 (d, J=6.4 Hz, 3H), 0.93 (d, J=6.4 Hz, 3H), 0.91 (t, J=7.6 Hz,
3H), 0.87 (d, J=6.8 Hz, 3H), 0.85 (d, J=6.8 Hz, 3H), 0.83 ppm
(t, J=7.6 Hz, 3H); 13C NMR (100 MHz, CDCl3): d=173.4, 173.1,
170.7, 170.6, 168.8, 168.6, 109.2, 71.6, 70.8, 69.1, 60.7, 58.6, 56.1,
53.5, 46.3, 39.2, 37.5, 34.5, 34.4, 33.4, 32.9, 30.9, 29.3, 28.7, 26.9,
25.7, 25.6, 25.5, 24.3, 23.8, 23.6, 22.5, 16.7, 15.4, 11.4, 10.7 ppm; IR
(neat): n˜ =3385, 3299, 2962, 2934, 2875, 1731, 1672, 1630, 1515,
1446, 1413, 1380, 1370, 1347, 1219, 1181, 1061, 752 cmÀ1; HRMS
(FAB): m/z calcd for C36H62N5O9 [M+H]+: 708.4548; found:
708.4543.
Compound 9l {1, 3, 2}: Yield: 41% from polymer-supported H-
bAla-OH 10 (31.6 mg, 43.1 mmol); [a]2D5 =À109 (c=0.825, CHCl3);
1H NMR (400 MHz, CDCl3): d=7.91 (d, J=8.0 Hz, 1H), 7.10–7.30 (m,
11 H), 5.47 (dd, J=4.0, 12.0 Hz, 1H), 5.22 (dt, J=4.4, 10.0 Hz, 1H),
5.04 (t, J=6.8 Hz, 1H), 4.84 (q, J=6.8 Hz, 1H), 4.63 (d, J=7.2 Hz,
1H), 4.06–4.15 (m, 2H), 3.93–4.01 (m, 1H), 3.58–3.71 (m, 3H), 3.41
(t, J=12.0 Hz, 1H), 3.35 (s, 3H), 3.07–3.13 (m, 1H), 3.04–3.07 (m,
1H), 2.89 (dd, J=3.6, 12.0 Hz, 1H), 2.81–2.86 (m, 1H), 2.64 (s, 3H),
2.51–2.63 (m, 2H), 2.15–2.22 (m, 1H), 2.08–2.14 (m, 2H), 1.70–1.83
(m, 2H), 1.42 (s, 3H), 1.32 (s, 3H), 1.17–1.28 (m, 1H), 0.25 ppm (d,
J=6.8 Hz, 3H); 13C NMR (100 MHz, CDCl3): d=173.4, 172.5, 170.5,
170.0, 169.7, 168.7, 136.3, 136.0, 129.4, 129.2, 128.8, 128.5, 127.3,
126.9, 109.1, 71.6, 70.7, 69.0, 60.4, 55.4, 53.8, 50.7, 46.3, 38.0, 36.6,
34.5, 34.3, 33.2, 31.0, 28.8, 28.0, 26.8, 25.4, 23.2, 13.2 ppm; IR (neat):
n˜ =3383, 3312, 2985, 2935, 2883, 1728, 1666, 1636, 1532, 1447,
1413, 1379, 1342, 1229, 1182, 1060, 752, 701 cmÀ1; HRMS (FAB): m/
z calcd for C39H52N5O9 [M+H]+: 734.3765; found: 734.3768.
Compound 9i {3, 3, 1}: Yield: 49% from polymer-supported H-
bAla-OH 10 (37.9 mg, 51.1 mmol); [a]2D4 =À143 (c=1.06, CHCl3);
1H NMR (400 MHz, CDCl3): d=8.32 (d, J=9.2 Hz, 1H), 7.15–7.31 (m,
5H), 7.14 (d, J=9.2 Hz, 1H), 5.48 (dd, J=5.6, 10.8 Hz, 1H), 5.06 (t,
J=8.0 Hz, 1H), 4.97 (dd, J=6.0, 9.2 Hz, 1H), 4.73 (d, J=6.4 Hz, 1H),
4.65 (dd, J=3.6, 6.8 Hz, 1H), 3.99–4.12 (m, 4H), 3.89 (t, J=7.6 Hz,
1H), 3.76–3.85 (m, 1H), 3.62 (t, J=6.4 Hz, 1H), 3.33–3.41 (m, 1H),
3.30 (s, 3H), 3.05 (brt, J=12.0 Hz, 1H), 2.92 (dd, J=4.0, 14.0 Hz,
1H), 2.67 (s, 3H), 2.48–2.57 (m, 3H), 2.10–2.22 (m, 2H), 2.05–2.10
(m, 1H), 1.92–1.98 (m, 2H), 1.81–1.91 (m, 1H), 1.72 (dt, J=5.2,
12.8 Hz, 1H), 1.42 (s, 3H), 1.41–1.45 (m, 1H), 1.33 (s, 3H), 1.30–1.37
(m, 1H), 1.21–1.30 (m, 1H), 0.88 (d, J=6.8 Hz, 3H), 0.84 (t, J=
7.2 Hz, 3H), 0.66 (d, J=6.8 Hz, 3H), 0.65 (d, J=6.4 Hz, 3H),
À0.31 ppm (ddd, J=3.6, 8.8, 12.8 Hz, 1H); 13C NMR (100 MHz,
CDCl3): d=173.3, 172.5, 170.6, 170.5, 168.8, 168.6, 136.0, 129.3,
128.9, 127.2, 109.2, 71.6, 70.7, 69.0, 60.6, 58.4, 53.4, 53.1, 46.3, 37.6,
37.2, 36.2, 34.5, 34.3, 32.8, 31.4, 29.4, 28.7, 26.8, 25.5, 24.7, 24.1,
23.8, 23.0, 21.8, 15.6, 11.4 ppm; IR (neat): n˜ =3384, 3307, 2960,
2936, 2875, 1731, 1670, 1636, 1518, 1448, 1412, 1380, 1370, 1338,
1218, 1182, 1061, 753, 702 cmÀ1; HRMS (FAB): m/z calcd for
C39H60N5O9 [M+H]+: 742.4391; found: 742.4374.
Compound 9m {2, 1, 2}: Yield: 16% from polymer-supported H-
bAla-OH 10 (12.0 mg, 16.8 mmol); [a]2D5 =À154 (c=0.525, CHCl3);
1H NMR (400 MHz, CDCl3): d=7.74 (d, J=10.0 Hz, 1H), 7.37 (d, J=
8.8 Hz, 1H), 7.13–7.30 (m, 5H), 5.25 (d, J=10.8 Hz, 1H), 5.11 (dd,
J=5.6, 8.8 Hz, 1H), 4.93 (ddd, J=4.4, 8.8, 11.2 Hz, 1H), 4.63 (d, J=
7.6 Hz, 1H), 4.46 (d, J=10.8 Hz, 1H), 4.05–4.18 (m, 3H), 3.59–3.75
(m, 3H), 3.24 (brt, J=12.8 Hz, 1H), 3.17 (s, 3H), 3.01 (s, 3H), 2.90–
2.97 (m, 2H), 2.75 (dd, J=5.6, 18.4 Hz, 1H), 2.56 (dd, J=10.0,
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18.4 Hz, 1H), 2.38–2.50 (m, 2H), 2.14–2.23 (m, 1H), 2.07–2.14 (m,
2H), 1.81–1.87 (m, 1H), 1.70–1.78 (m, 1H), 1.44 (s, 3H), 1.32 (s, 3H),
1.19–1.28 (m, 1H), 0.96 (d, J=6.4 Hz, 3H), 0.90 (d, J=6.4 Hz, 3H),
0.87 (d, J=6.4 Hz, 3H), 0.85 ppm (d, J=6.8 Hz, 3H); 13C NMR
(100 MHz, CDCl3): d=174.0, 172.8, 170.6, 170.2, 169.4, 168.2, 136.9,
128.8, 128.5, 126.9, 109.2, 71.5, 71.4, 69.2, 65.6, 60.4, 56.9, 51.8,
46.3, 36.4, 34.4, 34.2, 32.8, 29.7, 29.2, 28.8, 28.2, 27.5, 26.9, 25.5,
23.2, 20.2, 19.6, 19.2, 18.4 ppm; IR (CHCl3): n˜ =3389, 3301, 2960,
2936, 2873, 1727, 1673, 1635, 1530, 1446, 1413, 1380, 1370, 1221,
1190, 1062, 753, 700 cmÀ1; HRMS (FAB): m/z calcd for C37H56N5O9
[M+H]+: 714.4078; found: 714.4078.
d=173.7, 173.1, 170.4, 169.8, 168.8, 168.5, 136.6, 129.1, 128.4,
126.8, 109.2, 71.5, 71.0, 69.0, 60.2, 58.3, 57.7, 51.1, 46.1, 39.3, 37.5,
34.5, 34.2, 32.9, 30.3, 28.8, 28.5, 27.5, 26.8, 25.4, 25.3, 23.6, 23.0,
22.6, 19.8, 19.3 ppm; IR (neat): n˜ =3391, 3291, 2967, 2936, 2878,
1725, 1674, 1630, 1532, 1446, 1414, 1380, 1371, 1345, 1308, 1222,
1185, 1061, 751, 701 cmÀ1; HRMS (FAB): m/z calcd for C38H58N5O9
[M+H]+: 728.4235; found: 728.4221.
Compound 9q {3, 2, 2}: Yield: 48% from polymer-supported H-
bAla-OH 10 (37.4 mg, 50.4 mmol); [a]2D4 =À158 (c=0.675, CHCl3);
1H NMR (400 MHz, CDCl3): d=8.00 (d, J=9.6 Hz, 1H), 7.33 (d, J=
8.8 Hz, 1H), 7.21–7.27 (m, 5H), 5.14 (d, J=11.2 Hz, 1H), 5.09–5.13
(m, 1H), 5.08 (t, J=6.8 Hz, 1H), 5.03 (dd, J=3.6, 11.2 Hz, 1H), 4.64
(d, J=7.2 Hz, 1H), 4.12–4.20 (m, 1H), 4.04–4.11 (m, 2H), 3.60–3.69
(m, 3H), 3.22 (s, 3H), 3.18 (brt, J=12.0 Hz, 1H), 2.96–3.03 (m, 1H),
2.85 (s, 3H), 2.81–2.89 (m, 1H), 2.64–2.70 (m, 2H), 2.33 (dt, J=3.6,
11.2 Hz, 1H), 2.15–2.21 (m, 2H), 2.08–2.14 (m, 2H), 1.68–1.82 (m,
3H), 1.45–1.57 (m, 1H), 1.43 (s, 3H), 1.32 (s, 3H), 1.02–1.17 (m, 2H),
0.99 (d, J=6.4 Hz, 3H), 0.95 (d, J=6.8 Hz, 3H), 0.91 (t, J=7.2 Hz,
3H), 0.86 ppm (d, J=6.4 Hz, 3H); 13C NMR (100 MHz, CDCl3): d=
173.7, 173.1, 170.4, 170.0, 168.9, 168.5, 136.6, 129.1, 128.4, 126.9,
109.2, 71.6, 71.0, 69.0, 60.2, 58.3, 56.0, 51.1, 46.2, 39.3, 37.5, 34.5,
34.3, 33.5, 33.0, 30.4, 28.8, 28.5, 26.8, 25.5, 25.4, 25.3, 23.6, 23.1,
22.6, 16.0, 10.7 ppm; IR (neat): n˜ =3389, 3302, 2962, 2936, 2875,
1785, 1726, 1676, 1631, 1530, 1449, 1413, 1381, 1370, 1228, 1193,
1110, 1062, 752, 700 cmÀ1; HRMS (FAB): m/z calcd for C39H60N5O9
[M+H]+: 742.4391; found: 742.4401.
Compound 9n {2, 2, 2}: Yield: 10% from polymer-supported H-
bAla-OH 10 (7.6 mg, 10.5 mmol); [a]2D4 =À168 (c=0.210, CHCl3);
1H NMR (400 MHz, CDCl3): d=7.75 (d, J=10.0 Hz, 1H), 7.36 (d, J=
8.8 Hz, 1H), 7.22–7.30 (m, 5H), 5.29 (d, J=10.4 Hz, 1H), 5.11 (dd,
J=5.6, 8.0 Hz, 1H), 4.90–4.97 (m, 1H), 4.63 (d, J=8.0 Hz, 1H), 4.46
(d, J=10.8 Hz, 1H), 4.05–4.18 (m, 3H), 3.67–3.74 (m, 1H), 3.59–3.67
(m, 2H), 3.23 (brt, J=12.0 Hz, 1H), 3.16 (s, 3H), 3.01 (s, 3H), 2.85–
2.96 (m, 2H), 2.75 (dd, J=5.6, 18.8 Hz, 1H), 2.56 (dd, J=11.2,
18.8 Hz, 1H), 2.37–2.50 (m, 1H), 2.14–2.25 (m, 2H), 2.07–2.14 (m,
2H), 1.79–1.87 (m, 1H), 1.71–1.79 (m, 1H), 1.43 (s, 3H), 1.34–1.41
(m, 1H), 1.32 (s, 3H), 1.17–1.29 (m, 1H), 1.00–1.07 (m, 1H), 0.96 (d,
J=6.4 Hz, 3H), 0.90 (t, J=7.2 Hz, 3H), 0.86 (d, J=7.2 Hz, 3H),
0.84 ppm (d, J=6.8 Hz, 3H); 13C NMR (100 MHz, CDCl3): d=174.0,
172.8, 170.6, 170.3, 169.4, 168.3, 136.9, 128.9, 128.5, 126.9, 109.3,
71.5, 71.4, 69.2, 65.6, 60.4, 55.8, 51.8, 46.3, 36.5, 34.5, 34.3, 33.8,
32.8, 29.8, 29.2, 28.8, 28.3, 26.9, 25.5, 24.6, 23.2, 19.7, 19.2, 16.4,
11.2 ppm; IR (neat): n˜ =3392, 3286, 2967, 2934, 2877, 1725, 1675,
1629, 1532, 1446, 1414, 1380, 1223, 1184, 1119, 1061, 752,
Compound 9r {3, 3, 2}: Yield: 28% from polymer-supported H-
bAla-OH 10 (23.0 mg, 29.6 mmol); [a]2D6 =À124 (c=1.00, CHCl3);
1H NMR (400 MHz, CDCl3): d=8.02 (d, J=8.4 Hz, 1H), 7.14–7.32 (m,
11 H), 5.56 (dd, J=5.2, 6.8 Hz, 1H), 5.20 (ddd, J=4.4, 9.2, 10.8 Hz,
1H), 5.06 (t, J=7.2 Hz, 1H), 4.76 (q, J=4.0, 10.8 Hz, 1H), 4.65 (d,
J=7.2 Hz, 1H), 4.02–4.13 (m, 3H), 3.58–3.67 (m, 3H), 3.41 (t, J=
11.6 Hz, 1H), 3.32 (s, 3H), 3.08–3.13 (m, 1H), 3.04 (dd, J=4.4,
14.4 Hz, 1H), 2.87–2.93 (m, 1H), 2.78–2.86 (m, 1H), 2.74 (s, 3H),
2.53–2.64 (m, 2H), 2.15–2.20 (m, 1H), 2.04–2.14 (m, 2H), 1.68–1.81
(m, 3H), 1.43 (s, 3H), 1.32 (s, 3H), 1.20–1.28 (m, 1H), 1.08–1.20 (m,
1H), 0.68 (d, J=6.8 Hz, 1H), 0.67 (d, J=6.4 Hz, 3H), À0.23 ppm
(ddd, J=4.0, 8.8, 12.8 Hz, 1H); 13C NMR (100 MHz, CDCl3): d=173.5,
172.5, 170.2, 169.8, 168.7, 168.6, 136.4, 136.1, 129.3, 129.1, 128.9,
128.5, 127.2, 126.9, 109.2, 71.6, 70.8, 69.0, 60.2, 58.4, 53.1, 50.8,
46.2, 37.9, 37.4, 36.4, 34.6, 34.3, 32.8, 30.9, 28.8, 28.7, 26.8, 25.5,
24.8, 23.1, 23.0, 22.0 ppm; IR (neat): n˜ =3387, 3310, 2955, 2871,
1727, 1671, 1636, 1529, 1448, 1414, 1380, 1338, 1217, 1189, 1059,
753, 701 cmÀ1; HRMS (FAB): m/z calcd for C42H58N5O9 [M+H]+:
776.4235; found: 776.4227.
700 cmÀ1
;
HRMS (FAB): m/z calcd for C38H58N5O9 [M+H]+:
728.4235; found: 728.4255.
Compound 9o {2, 3, 2}: Yield: 24% from polymer-supported H-
bAla-OH 10 (19.2 mg, 25.2 mmol); [a]2D8 =À118 (c=0.845, CHCl3);
1H NMR (400 MHz, CDCl3): d=7.94 (d, J=9.2 Hz, 1H), 7.36 (d, J=
9.2 Hz, 1H), 7.18–7.28 (m, 10H), 5.83 (d, J=7.6 Hz, 1H), 5.07 (dd,
J=6.0, 7.6 Hz, 1H), 5.01–5.07 (m, 1H), 4.64 (d, J=7.2 Hz, 1H), 4.41
(d, J=10.8 Hz, 1H), 4.05–4.18 (m, 3H), 3.60–3.68 (m, 3H), 3.34 (dd,
J=7.6, 14.4 Hz, 1H), 3.23 (s, 3H), 3.07 (brt, J=14.0 Hz, 1H), 3.07
(dd, J=7.6, 14.4 Hz, 1H), 2.98–3.02 (m, 1H), 2.88 (s, 3H), 2.82–2.88
(m, 1H), 2.70 (dd, J=4.4, 18.8 Hz, 1H), 2.59 (dd, J=11.2, 18.8 Hz,
1H), 2.21–2.28 (m, 1H), 2.07–2.20 (m, 3H), 1.68–1.81 (m, 2H), 1.43
(s, 3H), 1.32 (s, 3H), 1.07–1.20 (m, 1H), 0.84 (d, J=6.4 Hz, 3H),
0.25 ppm (d, J=7.2 Hz, 3H); 13C NMR (100 MHz, CDCl3): d=173.8,
172.6, 170.5, 169.8, 168.9, 168.6, 136.6, 136.0, 129.1, 129.0, 128.8,
128.4, 127.1, 126.9, 109.2, 71.5, 71.2, 69.1, 66.0, 60.3, 51.7, 51.2,
46.2, 37.5, 35.7, 34.5, 34.3, 32.8, 30.5, 29.1, 28.6, 27.1, 26.8, 25.5,
23.1, 19.3, 17.7 ppm; IR (neat): n˜ =3388, 3299, 2963, 2932, 2875,
1726, 1672, 1631, 1530, 1446, 1413, 1380, 1337, 1221, 1189, 1055,
750, 700 cmÀ1; HRMS (FAB): m/z calcd for C41H56N5O9 [M+H]+:
762.4078; found: 762.4089.
General procedure II: Synthesis of destruxin E analogues 6
A 3m aqueous solution of HCl (0.500 mL) was added to a solution
of macrolactone 9 (1 equiv) in 1,4-dioxane (0.500 mL) and H2O
(0.500 mL). After being stirred at 08C for 1 h, the reaction mixture
was concentrated in vacuo, and the resulting residue was used in
the next reaction without further purification.
Compound 9p {3, 1, 2}: Yield: 49% from polymer-supported H-
bAla-OH 10 (38.3 mg, 51.6 mmol); [a]2D4 =À149 (c=0.800, CHCl3);
1H NMR (400 MHz, CDCl3): d=7.98 (d, J=9.6 Hz, 1H), 7.34 (d, J=
9.2 Hz, 1H), 7.15–7.31 (m, 5H), 4.98–5.18 (m, 4H), 4.64 (d, J=
7.6 Hz, 1H), 3.99–4.08 (m, 3H), 3.58–3.72 (m, 3H), 3.24 (s, 3H), 3.18
(brt, J=12.8 Hz, 1H), 2.93–3.08 (m, 1H), 2.85 (s, 3H), 2.78–2.85 (m,
1H), 2.58–2.75 (m, 2H), 2.28–2.43 (m, 2H), 2.08–2.20 (m, 3H), 1.66–
1.80 (m, 2H), 1.48–1.61 (m, 1H), 1.43 (s, 3H), 1.32 (s, 3H), 1.05–1.20
(m, 1H), 0.99 (d, J=6.4 Hz, 3H), 0.95 (d, J=7.2 Hz, 3H), 0.93 (d, J=
6.4 Hz, 3H), 0.90 ppm (d, J=6.0 Hz, 3H); 13C NMR (100 MHz, CDCl3):
TsCl (2.00 equiv) was added to a solution of the crude diol
(1 equiv), triethylamine (5.00 equiv), and DMAP (0.100 equiv) in dry
CH2Cl2 (1.00 mL) at 08C under an argon atmosphere. After being
stirred at room temperature for 12 h, the reaction mixture was
poured into a saturated aqueous solution of NH4Cl and aqueous
layer was extracted with CHCl3. The organic layer was washed with
a 1m aqueous solution of HCl, a saturated aqueous solution of
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NaHCO3, and brine. The organic layer was dried over MgSO4 and
filtered. The filtrate was concentrated in vacuo, and the resulting
residue was purified by flash column chromatography on silica gel
(eluted with CHCl3/MeOH=40:1) to afford tosylate S3 as a colorless
oil. (Data for S3 are shown in the Supporting Information.)
(150 MHz, CDCl3): d=173.3, 172.6, 170.9, 170.7, 169.8, 168.7, 136.0,
129.5, 128.9, 127.3, 70.5, 60.9, 55.7, 54.0, 53.3, 47.9, 47.1, 46.7, 37.7,
36.6, 34.5, 33.7, 33.2, 31.6, 29.5, 28.1, 24.3, 24.0, 15.6, 13.1,
11.4 ppm; IR (neat): n˜ =2961, 2926, 2875, 2849, 1731, 1666, 1634,
1519, 1447, 1380, 1179, 1130, 1102 cmÀ1; HRMS (FAB): m/z calcd for
C33H48N5O8 [M+H]+: 642.3503; found: 642.3511.
K2CO3 (4.00 equiv) was added to a solution of tosylate S3 (1 equiv)
in iPrOH/(CH2Cl)2 (10:1, 2.00 mL) at 08C under an argon atmos-
phere. After being stirred at 508C for 12 h, the reaction mixture
was poured into a saturated aqueous solution of NH4Cl and the
aqueous layer was extracted with CHCl3. The organic layer was
washed with brine, dried over MgSO4, and filtered. The filtrate was
concentrated in vacuo, and the resulting residue was purified by
flash column chromatography on silica gel (eluted with CHCl3/
MeOH=40:1) to afford destruxin E derivatives 6 as a colorless oil.
Compound 6d {2, 1, 1}: Yield (3 steps): 21% (2.0 mg, 3.22 mmol);
[a]2D5 =À233 (c=0.100, CHCl3); 1H NMR (400 MHz, CDCl3): d=8.12
(d, J=9.6 Hz, 1H), 7.16 (d, J=8.8 Hz, 1H), 5.24 (d, J=10.8 Hz, 1H),
5.05 (t, J=7.2 Hz, 1H), 4.69 (d, J=8.4 Hz, 1H), 4.66 (t, J=8.8 Hz,
1H), 4.38 (d, J=10.8 Hz, 1H), 3.93–4.06 (m, 2H), 3.62–3.70 (m, 1H),
3.15 (s, 3H), 3.08–3.14 (m, 1H), 2.96–3.00 (m, 1H), 2.95 (s, 3H), 2.84
(t, J=4.0 Hz, 1H), 2.61 (dd, J=5.2, 18.0 Hz, 1H), 2.57–2.70 (m, 1H),
2.49–2.57 (m, 1H), 2.37–2.48 (m, 2H), 2.27–2.36 (m, 2H), 2.07–2.13
(m, 2H), 1.93–2.04 (m, 3H), 1.46–1.54 (m, 1H), 1.22–1.34 (m, 1H),
0.94 (d, J=6.4 Hz, 3H), 0.90 (d, J=6.4 Hz, 3H), 0.89 (d, J=5.6 Hz,
3H), 0.88 (d, J=7.6 Hz, 3H), 0.84 (d, J=6.8 Hz, 3H), 0.78 ppm (d,
J=7.2 Hz, 3H); 13C NMR (150 MHz, CDCl3): d=173.6, 173.2, 171.2,
170.6, 169.4, 168.2, 71.2, 65.9, 60.8, 56.7, 53.7, 47.9, 47.2, 46.6, 37.3,
34.1, 33.6, 32.9, 30.5, 29.2, 29.1, 28.6, 27.2, 25.7, 24.1, 20.4, 19.8,
19.3, 18.7, 14.4, 11.0 ppm; IR (neat): n˜ =2966, 2930, 2877, 1729,
Compound 6a {1, 1, 1} (destruxin E): Yield (3 steps): 34% (8.1 mg,
13.6 mmol); [a]2D8 =À224 (c=0.200, CHCl3); 1H NMR (400 MHz,
CDCl3): d=8.20 (d, J=8.0 Hz, 1H), 7.15 (d, J=9.2 Hz, 1H), 5.16 (q,
J=6.6 Hz, 1H), 4.99 (t, J=7.3 Hz, 1H), 4.97 (d, J=10.9 Hz, 1H), 4.90
(dd, J=6.6, 9.1 Hz, 1H), 4.69 (d, J=7.0 Hz, 1H), 4.01–4.07 (m, 1H),
3.95 (brt, J=8.3 Hz, 1H), 3.60–3.67 (m, 1H), 3.23 (s, 3H), 3.07 (brt,
J=12.8 Hz, 1H), 2.93–2.97 (m, 1H), 2.83 (t, J=4.6 Hz, 1H), 2.72 (s,
3H), 2.63–2.68 (m, 1H), 2.47–2.59 (m, 3H), 2.24–2.35 (m, 2H), 1.81–
2.07 (m, 5H), 1.38–1.48 (m, 2H), 1.30 (d, J=6.8 Hz, 3H), 1.27–1.31
(m, 1H), 0.92 (d, J=6.6 Hz, 3H), 0.89 (d, J=6.8 Hz, 3H), 0.86 (d, J=
7.2 Hz, 3H), 0.85 ppm (t, J=7.8 Hz, 3H); 13C NMR (100 MHz, CDCl3):
d=173.5, 173.4, 171.0, 170.8, 169.7, 168.6, 70.6, 60.9, 58.0, 55.5,
53.6, 47.8, 47.1, 46.6, 37.4, 34.5, 33.6, 33.1, 31.2, 30.8, 29.6, 29.1,
28.1, 27.2, 24.3, 24.0, 20.0, 19.6, 15.4, 15.1, 11.3 ppm; IR (neat): n˜ =
2965, 2929, 1734, 1670, 1629, 1518, 1444, 1179 cmÀ1; HRMS (FAB):
m/z calcd for C29H48N5O8 [M+H]+: 594.3497; found: 594.3480.
1671, 1625, 1523, 1448, 1383, 1350, 1224, 1180, 1119, 1077 cmÀ1
;
HRMS (FAB): m/z calcd for C31H52N5O8 [M+H]+: 622.3816; found:
622.3822.
Compound 6e {2, 2, 1}: Yield (3 steps): 28% (1.3 mg, 2.04 mmol);
1
[a]2D5 =À250 (c=0.0500, CHCl3); H NMR (400 MHz, CDCl3): d=8.14
(d, J=10.0 Hz, 1H), 7.16 (d, J=8.8 Hz, 1H), 5.29 (d, J=10.8 Hz, 1H),
5.04 (dd, J=6.4, 7.6 Hz, 1H), 4.70 (d, J=7.2 Hz, 1H), 4.66 (t, J=
8.8 Hz, 1H), 4.37 (d, J=10.4 Hz, 1H), 3.93–4.06 (m, 2H), 3.62–3.70
(m, 1H), 3.13 (s, 3H), 3.06–3.12 (m, 1H), 2.96–3.00 (m, 1H), 2.95 (s,
3H), 2.84 (t, J=4.4 Hz, 1H), 2.61 (dd, J=4.4, 18.4 Hz, 1H), 2.51–2.60
(m, 2H), 2.35–2.47 (m, 2H), 2.29–2.34 (m, 1H), 2.12–2.21 (m, 1H),
2.03–2.10 (m, 2H), 1.92–2.02 (m, 3H), 1.44–1.54 (m, 1H), 1.22–1.43
(m, 2H), 0.97–1.08 (m, 1H), 0.94 (d, J=6.4 Hz, 3H), 0.90 (t, J=
7.6 Hz, 3H), 0.87 (d, J=7.6 Hz, 3H), 0.86 (t, J=7.6 Hz, 3H), 0.85 (d,
J=6.8 Hz, 3H), 0.78 ppm (d, J=7.2 Hz, 3H); 13C NMR (150 MHz,
CDCl3): d=173.6, 173.2, 171.2, 170.7, 169.4, 168.3, 71.2, 65.9, 60.8,
55.6, 53.7, 47.9, 47.1, 46.6, 37.4, 34.1, 33.6, 33.5, 32.9, 30.6, 29.2,
29.1, 28.7, 25.7, 24.8, 24.1, 19.9, 19.4, 16.7, 14.4, 11.1, 11.0 ppm; IR
(neat): n˜ =2964, 2929, 2879, 1728, 1671, 1625, 1523, 1445, 1382,
1225, 1179, 1077 cmÀ1; HRMS (FAB): m/z calcd for C32H54N5O8 [M+
H]+: 636.3972; found: 636.3965.
Compound 6b {1, 2, 1}: Yield (3 steps): 45% (7.8 mg, 12.8 mmol);
[a]2D5 =À211 (c=0.110, CHCl3); 1H NMR (400 MHz, CDCl3): d=8.22
(d, J=8.8 Hz, 1H), 7.14 (d, J=9.2 Hz, 1H), 5.13 (q, J=6.8 Hz, 1H),
5.04 (d, J=11.2 Hz, 1H), 4.99 (t, J=6.8 Hz, 1H), 4.88 (dd, J=6.8,
10.0 Hz, 1H), 4.69 (d, J=6.8 Hz, 1H), 4.00–4.08 (m, 1H), 3.95 (t, J=
8.0 Hz, 1H), 3.62–3.70 (m, 1H), 3.21 (s, 3H), 3.08 (brt, J=12.8 Hz,
1H), 2.93–2.98 (m, 1H), 2.83 (t, J=4.8 Hz, 1H), 2.72 (s, 3H), 2.63–
2.68 (m, 1H), 2.51–2.59 (m, 2H), 2.43–2.49 (m, 1H), 2.24–2.32 (m,
1H), 2.03–2.13 (m, 2H), 1.91–2.01 (m, 3H), 1.86–1.91 (m, 1H), 1.50–
1.76 (m, 2H), 1.38–1.48 (m, 2H), 1.31 (d, J=6.8 Hz, 3H), 0.92 (d, J=
5.6 Hz, 3H), 0.87 (t, J=6.8 Hz, 3H), 0.86 (d, J=6.8 Hz, 3H),
0.85 ppm (t, J=6.8 Hz, 3H); 13C NMR (100 MHz, CDCl3): d=173.6,
173.5, 171.1, 170.9, 169.7, 168.6, 70.7, 60.9, 56.8, 55.5, 53.7, 47.9,
47.1, 46.7, 37.5, 34.5, 33.7, 33.5, 33.2, 31.0, 29.2, 28.1, 25.8, 24.5,
24.0, 16.2, 15.4, 15.2, 11.4, 11.1 ppm; IR (neat): n˜ =2964, 2930, 2876,
1732, 1668, 1633, 1519, 1444, 1380, 1340, 1280, 1257, 1127,
Compound 6 f {2, 3, 1}: Yield (3 steps): 26% (4.7 mg, 7.07 mmol);
[a]2D4 =À202 (c=0.155, CHCl3); 1H NMR (400 MHz, CDCl3): d=8.35
(d, J=9.6 Hz, 1H), 7.16–7.31 (m, 5H), 7.16 (d, J=9.2 Hz, 1H), 5.81
(dd, J=6.8, 8.8 Hz, 1H), 5.01 (d, J=6.8 Hz, 1H), 4.84 (dd, J=6.4,
9.2 Hz, 1H), 4.72 (d, J=6.8 Hz, 1H), 4.36 (d, J=10.4 Hz, 1H), 4.06–
4.17 (m, 1H), 3.94 (t, J=8.0 Hz, 1H), 3.62–3.70 (m, 1H), 3.20–3.29
(m, 2H), 3.19 (s, 3H), 3.08–3.16 (m, 1H), 2.94–3.02 (m, 1H), 2.84 (t,
J=4.4 Hz, 1H), 2.80 (s, 3H), 2.60–2.68 (m, 2H), 2.57–2.60 (m, 1H),
2.43–2.53 (m, 1H), 2.22–2.36 (m, 2H), 2.03–2.12 (m, 1H), 1.93–2.02
(m, 3H), 1.82–1.91 (m, 1H), 1.36–1.47 (m, 1H), 1.20–1.34 (m, 1H),
0.88 (d, J=6.4 Hz, 3H), 0.84 (t, J=7.2 Hz, 3H), 0.84 (d, J=6.8 Hz,
3H), 0.49 ppm (d, J=7.2 Hz, 3H); 13C NMR (150 MHz, CDCl3): d=
173.4, 172.9, 171.5, 170.5, 168.7, 168.6, 136.1, 128.9, 128.6, 127.1,
70.8, 66.3, 60.8, 53.5, 51.4, 47.9, 47.1, 46.6, 37.6, 35.0, 34.5, 33.6,
32.8, 31.2, 29.2, 29.0, 27.2, 24.4, 24.0, 19.7, 18.5, 15.4, 11.3 ppm; IR
(neat): n˜ =2964, 2929, 2876, 1731, 1668, 1630, 1519, 1446, 1383,
1180, 1078 cmÀ1; HRMS (FAB): m/z calcd for C35H52N5O8 [M+H]+:
670.3816; found: 670.3815.
1101 cmÀ1
;
HRMS (FAB): m/z calcd for C30H50N5O8 [M+H]+:
608.3659; found: 608.3657.
Compound 6c {1, 3, 1}: Yield (3 steps): 30% (5.2 mg, 8.18 mmol);
[a]2D5 =À166 (c=0.115, CHCl3); 1H NMR (400 MHz, CDCl3): d=8.26
(d, J=8.0 Hz, 1H), 7.15–7.36 (m, 5H), 7.11 (d, J=9.2 Hz, 1H), 5.39
(dd, J=4.0, 12.0 Hz, 1H), 4.93–5.05 (m, 2H), 4.70 (d, J=7.6 Hz, 1H),
4.68 (q, J=7.2 Hz, 1H), 3.88–4.00 (m, 2H), 3.62–3.70 (m, 1H), 3.36
(s, 3H), 3.32–3.39 (m, 1H), 3.04 (brt, J=11.2 Hz, 1H), 2.94–2.98 (m,
1H), 2.92 (dd, J=4.0, 12.0 Hz, 1H), 2.83 (t, J=4.0 Hz, 1H), 2.58–2.68
(m, 1H), 2.56 (s, 3H), 2.49–2.56 (m, 2H), 2.43–2.49 (m, 1H), 2.24–
2.32 (m, 1H), 2.04–2.10 (m, 1H), 1.91–2.01 (m, 3H), 1.82–1.90 (m,
1H), 1.38–1.48 (m, 1H), 1.25–1.34 (m, 1H), 0.90 (d, J=6.8 Hz, 3H),
0.86 (d, J=7.2 Hz, 3H), 0.18 ppm (d, J=7.2 Hz, 3H); 13C NMR
Chem. Eur. J. 2015, 21, 18417 – 18430
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Compound 6g {3, 1, 1}: Yield (3 steps): 15% (2.4 mg, 3.70 mmol);
(d, J=7.6 Hz, 1H), 7.35 (d, J=8.8 Hz, 1H), 7.21–7.30 (m, 5H), 5.31
(q, J=6.8 Hz, 1H), 5.10 (dt, J=4.0, 8.8 Hz, 1H), 5.02 (d, J=12.0 Hz,
1H), 5.01 (t, J=6.8 Hz, 1H), 4.63 (d, J=8.0 Hz, 1H), 4.05–4.13 (m,
1H), 3.72 (t, J=8.8 Hz, 1H), 3.45–3.53 (m, 1H), 3.23 (s, 3H), 3.16
(brt, J=12.4 Hz, 1H), 3.04 (dd, J=4.0, 13.6 Hz, 1H), 2.96–3.01 (m,
1H), 2.81 (s, 3H), 2.81–2.87 (m, 2H), 2.57 (dd, J=2.4, 4.8 Hz, 1H),
2.30–2.40 (m, 1H), 2.21–2.27 (m, 1H), 2.16–2.21 (m, 1H), 1.98–2.07
(m, 1H), 1.73–1.83 (m, 2H), 1.32 (d, J=6.8 Hz, 3H), 1.17–1.25 (m,
1H), 0.93 (d, J=6.4 Hz, 3H), 0.88 ppm (d, J=6.4 Hz, 3H); 13C NMR
(150 MHz, CDCl3): d=173.6, 173.5, 170.7, 170.1, 169.7, 168.4, 136.6,
129.1, 128.5, 126.9, 70.8, 60.4, 58.0, 55.3, 51.2, 47.9, 47.1, 46.5, 37.6,
34.6, 33.5, 33.2, 30.3, 28.5, 28.1, 27.4, 23.3, 19.8, 19.3, 15.2 ppm; IR
(neat): n˜ =2965, 2931, 2875, 1728, 1671, 1631, 1529, 1444, 1379,
1342, 1274, 1227, 1181, 1100 cmÀ1; HRMS (FAB): m/z calcd for
C32H46N5O8 [M+H]+: 628.3346; found: 628.3348.
1
[a]2D5 =À236 (c=0.0950, CHCl3); H NMR (400 MHz, CDCl3): d=8.33
(d, J=9.6 Hz, 1H), 7.18 (d, J=9.6 Hz, 1H), 5.01 (dd, J=4.4, 6.8 Hz,
1H), 4.99 (d, J=8.0 Hz, 1H), 4.93 (dd, J=6.4, 9.6 Hz, 1H), 4.90 (d,
J=10.0 Hz, 1H), 4.73 (d, J=6.8 Hz, 1H), 4.09–4.18 (m, 1H), 3.92 (t,
J=8.8 Hz, 1H), 3.62–3.70 (m, 1H), 3.22 (s, 3H), 3.05–3.13 (m, 1H),
2.94–3.00 (m, 1H), 2.84 (t, J=4.0 Hz, 1H), 2.76 (s, 3H), 2.59–2.64 (m,
2H), 2.57–2.70 (m, 1H), 2.49–2.57 (m, 1H), 2.57 (dd, J=2.4, 4.0 Hz,
1H), 2.51 (d, J=6.4 Hz, 1H), 2.26–2.42 (m, 2H), 2.04–2.12 (m, 1H),
1.98–2.02 (m, 2H), 1.93–1.98 (m, 2H), 1.83–1.92 (m, 1H), 1.47–1.58
(m, 1H), 1.35–1.43 (m, 1H), 1.22–1.33 (m, 1H), 1.00–1.06 (m, 1H),
0.97 (d, J=6.4 Hz, 3H), 0.93 (d, J=6.8 Hz, 6H), 0.91 (d, J=6.4 Hz,
3H), 0.84 (d, J=7.2 Hz, 3H), 0.83 ppm (t, J=7.6 Hz, 3H); 13C NMR
(150 MHz, CDCl3): d=173.4, 173.2, 170.7, 170.6, 168.7, 168.6, 70.8,
60.8, 58.7, 57.8, 53.6, 47.9, 47.0, 46.6, 39.3, 37.6, 34.5, 33.6, 32.9,
30.9, 29.2, 28.7, 27.4, 25.6, 24.3, 23.9, 23.6, 22.6, 20.1, 19.7, 15.5,
11.4 ppm; IR (neat): n˜ =2962, 2932, 2875, 1731, 1671, 1630, 1518,
1445, 1382, 1345, 1217, 1181, 1111, 1082 cmÀ1; HRMS (FAB): m/z
calcd for C32H54N5O8 [M+H]+: 636.3972; found: 636.3963.
Compound 6k {1, 2, 2}: Yield (3 steps): 39% (8.6 mg, 13.4 mmol);
[a]2D9 =À211 (c=0.225, CHCl3); 1H NMR (600 MHz, CDCl3): d=7.86
(d, J=8.4 Hz, 1H), 7.33 (d, J=7.8 Hz, 1H), 7.21–7.30 (m, 5H), 5.30
(q, J=7.2 Hz, 1H), 5.09 (d, J=10.8 Hz, 1H), 5.06–5.12 (m, 1H), 5.01
(t, J=7.2 Hz, 1H), 4.62 (d, J=8.4 Hz, 1H), 4.06–4.13 (m, 1H), 3.72 (t,
J=7.8 Hz, 1H), 3.45–3.51 (m, 1H), 3.21 (s, 3H), 3.15 (brt, J=
12.6 Hz, 1H), 3.02 (dd, J=4.2, 13.8 Hz, 1H), 2.99 (dt, J=4.2, 7.2 Hz,
1H), 2.81–2.86 (m, 2H), 2.81 (s, 3H), 2.71 (ddd, J=1.8, 12.0, 19.2 Hz,
1H), 2.62 (dd, J=2.4, 19.2 Hz, 1H), 2.57 (dd, J=2.4, 4.8 Hz, 1H),
2.23 (ddd, J=4.2, 7.2, 14.4 Hz, 1H), 2.16–2.21 (m, 1H), 2.07–2.13
(m, 1H), 2.02 (dt, J=7.2, 14.4 Hz, 1H), 1.74–1.82 (m, 2H), 1.43–1.49
(m, 1H), 1.32 (d, J=7.2 Hz, 3H), 1.18–1.27 (m, 1H), 0.96–1.04 (m,
1H), 0.92 (t, J=7.2 Hz, 3H), 0.85 ppm (d, J=6.6 Hz, 3H); 13C NMR
(150 MHz, CDCl3): d=173.7, 173.5, 170.7, 170.1, 169.7, 168.4, 136.6,
129.1, 128.5, 126.9, 70.8, 60.4, 56.6, 55.3, 51.2, 47.9, 47.1, 46.5, 37.5,
34.5, 33.6, 33.5, 33.2, 30.4, 28.5, 28.1, 25.5, 25.3, 16.0, 15.2,
11.0 ppm; IR (neat): n˜ =2965, 2933, 2879, 1728, 1671, 1631, 1529,
1444, 1380, 1227, 1181 cmÀ1; HRMS (FAB): m/z calcd for C33H48N5O8
[M+H]+: 642.3503; found: 642.3519.
Compound 6h {3, 2, 1}: Yield (3 steps): 26% (5.6 mg, 8.64 mmol);
[a]2D6 =À223 (c=0.185, CHCl3); 1H NMR (600 MHz, CDCl3): d=8.35
(d, J=10.2 Hz, 1H), 7.16 (d, J=9.6 Hz, 1H), 5.09 (d, J=11.4 Hz, 1H),
5.01 (t, J=7.2 Hz, 1H), 4.92 (dd, J=6.6, 9.6 Hz, 1H), 4.90 (d, J=
9.0 Hz, 1H), 4.73 (d, J=6.6 Hz, 1H), 4.07–4.16 (m, 1H), 3.94 (t, J=
8.4 Hz, 1H), 3.60–3.72 (m, 1H), 3.20 (s, 3H), 3.08 (brt, J=12.0 Hz,
1H), 2.94–3.00 (m, 1H), 2.84 (t, J=4.8 Hz, 1H), 2.77 (s, 3H), 2.59–
2.64 (m, 2H), 2.57 (dd, J=3.6, 4.8 Hz, 1H), 2.50 (d, J=5.4 Hz, 1H),
2.38 (dt, J=4.2, 12.6 Hz, 1H), 2.27–2.33 (m, 1H), 2.03–2.15 (m, 2H),
1.96–2.00 (m, 1H), 1.92–1.96 (m, 2H), 1.85–1.92 (m, 1H), 1.47–1.55
(m, 1H), 1.35–1.47 (m, 2H), 1.22–1.33 (m, 1H), 0.97–1.05 (m, 2H),
0.97 (d, J=6.6 Hz, 3H), 0.93 (d, J=6.6 Hz, 3H), 0.91 (t, J=7.2 Hz,
3H), 0.87 (d, J=6.6 Hz, 3H), 0.85 (d, J=6.6 Hz, 3H), 0.84 ppm (t, J=
7.2 Hz, 3H); 13C NMR (150 MHz, CDCl3): d=173.4, 173.1, 170.7,
170.6, 168.7, 168.6, 70.7, 60.8, 58.7, 56.2, 53.6, 47.9, 47.0, 46.5, 39.3,
37.6, 34.5, 33.6, 33.5, 32.9, 31.0, 29.2, 28.7, 25.7, 25.6, 24.3, 23.9,
23.6, 22.5, 16.2, 15.4, 11.4, 10.8 ppm; IR (neat): n˜ =2962, 2932,
2875, 1731, 1671, 1629, 1517, 1445, 1382, 1180, 1083 cmÀ1; HRMS
(FAB): m/z calcd for C33H56N5O8 [M+H]+: 650.4129; found:
650.4141.
Compound 6l {1, 3, 2}: Yield (3 steps): 18% (5.2 mg, 7.64 mmol);
[a]2D9 =À197 (c=0.115, CHCl3); 1H NMR (400 MHz, CDCl3): d=7.91
(d, J=7.2 Hz, 1H), 7.14–7.30 (m, 11 H), 5.46 (dd, J=4.0, 12.0 Hz,
1H), 5.21 (dt, J=4.4, 10.0 Hz, 1H), 4.98 (t, J=6.8 Hz, 1H), 4.85 (q,
J=6.8 Hz, 1H), 4.63 (d, J=8.0 Hz, 1H), 3.94–4.04 (m, 1H), 3.72 (t,
J=8.8 Hz, 1H), 3.48–3.55 (m, 1H), 3.40 (t, J=12.0 Hz, 1H), 3.34 (s,
3H), 3.04–3.13 (m, 2H), 2.95–3.01 (m, 1H), 2.89 (dd, J=3.6, 12.0 Hz,
1H), 2.79–2.86 (m, 3H), 2.64 (s, 3H), 2.52–2.60 (m, 2H), 2.17–2.26
(m, 2H), 2.00 (dt, J=3.2, 14.0 Hz, 1H), 1.73–1.85 (m, 2H), 1.16–1.25
(m, 1H), 0.25 ppm (d, J=6.4 Hz, 3H); 13C NMR (150 MHz, CDCl3):
d=173.5, 172.6, 170.5, 169.9, 169.7, 168.5, 136.3, 136.1, 129.4,
129.2, 128.9, 128.5, 127.3, 127.0, 70.6, 60.5, 55.4, 53.8, 50.7, 47.9,
47.0, 46.5, 38.0, 36.7, 34.5, 33.5, 33.2, 31.0, 28.7, 28.1, 23.3,
13.2 ppm; IR (neat): n˜ =2928, 1729, 1668, 1636, 1529, 1446, 1378,
1345, 1179, 1108, 1079 cmÀ1; HRMS (FAB): m/z calcd for C36H46N5O8
[M+H]+: 676.3346; found: 676.3364.
Compound 6i {3, 3, 1}: Yield (3 steps): 11 % (3.9 mg, 5.66 mmol);
[a]2D3 =À251 (c=0.110, CHCl3); 1H NMR (600 MHz, CDCl3): d=8.32
(d, J=9.0 Hz, 1H), 7.15–7.30 (m, 5H), 7.14 (d, J=9.0 Hz, 1H), 5.47
(dd, J=5.4, 10.2 Hz, 1H), 4.99 (t, J=7.2 Hz, 1H), 4.96 (dd, J=6.6,
9.0 Hz, 1H), 4.74 (d, J=6.6 Hz, 1H), 4.66 (dd, J=3.6, 10.8 Hz, 1H),
4.02–4.07 (m, 1H), 3.93 (t, J=9.0 Hz, 1H), 3.62–3.68 (m, 1H), 3.37 (t,
J=11.4 Hz, 1H), 3.30 (s, 3H), 3.03 (brt, J=10.8 Hz, 1H), 2.91–3.00
(m, 2H), 2.83 (t, J=4.2 Hz, 1H), 2.67 (s, 3H), 2.49–2.58 (m, 3H),
2.27–2.32 (m, 1H), 2.03–2.13 (m, 2H), 1.96–2.00 (m, 1H), 1.91–1.96
(m, 2H), 1.83–1.90 (m, 1H), 1.73 (dt, J=5.4, 12.6 Hz, 1H), 1.38–1.46
(m, 1H), 1.21–1.32 (m, 2H), 0.88 (d, J=6.6 Hz, 3H), 0.84 (t, J=
7.2 Hz, 3H), 0.67 (d, J=6.0 Hz, 3H), 0.65 (d, J=6.0 Hz, 3H),
À0.31 ppm (ddd, J=3.6, 9.0, 12.6 Hz, 1H); 13C NMR (150 MHz,
CDCl3): d=173.3, 172.6, 170.6, 170.5, 168.7, 168.6, 136.1, 129.3,
128.9, 127.2, 70.6, 60.8, 58.5, 53.5, 53.2, 47.9, 47.0, 46.6, 37.7, 37.3,
36.3, 34.6, 33.5, 32.8, 31.5, 29.4, 28.7, 24.8, 24.2, 23.9, 23.0, 21.8,
15.6, 11.4 ppm; IR (neat): n˜ =2958, 2925, 2875, 1731, 1670, 1630,
1518, 1448, 1382, 1341, 1180, 1076 cmÀ1; HRMS (FAB): m/z calcd for
C36H54N5O8 [M+H]+: 684.3972; found: 684.3964.
Compound 6m {2, 1, 2}: Yield (3 steps): 18% (2.0 mg, 3.06 mmol);
1
[a]2D7 =À211 (c=0.0900, CHCl3); H NMR (600 MHz, CDCl3): d=7.74
(d, J=9.6 Hz, 1H), 7.36 (d, J=8.4 Hz, 1H), 7.18–7.30 (m, 5H), 5.24
(d, J=10.2 Hz, 1H), 5.06 (t, J=7.2 Hz, 1H), 4.91–4.96 (m, 1H), 4.64
(d, J=7.8 Hz, 1H), 4.47 (d, J=10.8 Hz, 1H), 4.10–4.18 (m, 1H), 3.76
(t, J=9.0 Hz, 1H), 3.45–3.54 (m, 1H), 3.24 (brt, J=11.4 Hz, 1H), 3.16
(s, 3H), 3.01 (s, 3H), 2.93–3.01 (m, 2H), 2.84 (t, J=4.2 Hz, 1H), 2.75
(dd, J=5.4, 18.6 Hz, 1H), 2.56–2.62 (m, 2H), 2.37–2.47 (m, 2H),
2.27–2.33 (m, 1H), 2.08–2.14 (m, 1H), 1.98–2.04 (m, 1H), 1.81–1.86
(m, 1H), 1.72–1.81 (m, 1H), 1.21–1.29 (m, 1H), 0.97 (d, J=6.6 Hz,
Compound 6j {1, 1, 2}: Yield (3 steps): 27% (6.1 mg, 9.66 mmol);
[a]2D7 =À126 (c=0.185, CHCl3); 1H NMR (400 MHz, CDCl3): d=7.86
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3H), 0.90 (d, J=6.6 Hz, 3H), 0.87 (d, J=7.2 Hz, 3H), 0.86 ppm (d,
J=8.4 Hz, 3H); 13C NMR (150 MHz, CDCl3): d=174.0, 172.8, 170.5,
170.3, 169.4, 168.0, 136.9, 128.9, 128.5, 127.0, 71.4, 65.6, 60.5, 57.0,
51.9, 47.9, 47.1, 46.5, 36.5, 34.2, 33.5, 32.8, 29.7, 29.2, 28.7, 28.3,
27.5, 23.3, 20.2, 19.7, 19.2, 18.5 ppm; IR (neat): n˜ =2967, 2931,
2879, 1725, 1673, 1628, 1531, 1445, 1384, 1338, 1260, 1223, 1184,
33.4, 33.0, 30.3, 28.8, 28.4, 27.5, 25.4, 23.6, 23.2, 22.7, 19.9,
19.4 ppm; IR (neat): n˜ =2959, 2924, 2873, 1727, 1674, 1634, 1531,
1446, 1383, 1343, 1275, 1182, 1081 cmÀ1; HRMS (FAB): m/z calcd for
C35H52N5O8 [M+H]+: 670.3816; found: 670.3818.
Compound 6q {3, 2, 2}: Yield (3 steps): 10% (3.6 mg, 5.20 mmol);
1079 cmÀ1
;
HRMS (FAB): m/z calcd for C34H50N5O8 [M+H]+:
1
[a]2D3 =À194 (c=0.0900, CHCl3); H NMR (600 MHz, CDCl3): d=7.99
656.3659; found: 656.3655.
(d, J=9.6 Hz, 1H), 7.33 (d, J=8.4 Hz, 1H), 7.21–7.30 (m, 5H), 5.14
(d, J=10.8 Hz, 1H), 5.10–5.14 (m, 1H), 5.05 (dd, J=3.6, 11.4 Hz,
1H), 5.02 (t, J=5.4 Hz, 1H), 4.65 (d, J=7.8 Hz, 1H), 4.12–4.17 (m,
1H), 3.66 (t, J=9.6 Hz, 1H), 3.44–3.50 (m, 1H), 3.21 (s, 3H), 3.19
(brt, J=12.6 Hz, 1H), 2.97–3.04 (m, 2H), 2.85 (s, 3H), 2.84 (t, J=
4.8 Hz, 1H), 2.80–2.84 (m, 1H), 2.62–2.75 (m, 2H), 2.57 (dd, J=2.4,
4.8 Hz, 1H), 2.33 (dt, J=4.8, 12.0 Hz, 1H), 2.23–2.28 (m, 1H), 2.13–
2.21 (m, 2H), 1.99–2.05 (m, 1H), 1.71–1.79 (m, 2H), 1.48–1.55 (m,
1H), 1.42–1.47 (m, 1H), 1.20–1.27 (m, 1H), 1.00–1.17 (m, 2H), 0.99
(d, J=6.6 Hz, 3H), 0.95 (d, J=6.6 Hz, 3H), 0.91 (t, J=7.2 Hz, 3H),
0.86 ppm (d, J=6.6 Hz, 3H); 13C NMR (150 MHz, CDCl3): d=173.7,
173.1, 170.4, 170.0, 168.9, 168.4, 136.7, 129.1, 128.5, 126.9, 70.9,
60.3, 58.4, 56.1, 51.2, 48.0, 47.0, 46.4, 39.3, 37.5, 34.5, 33.5, 33.4,
33.0, 30.4, 28.8, 28.4, 25.4, 25.3, 23.6, 23.2, 22.6, 16.0, 10.7 ppm; IR
(neat): n˜ =2960, 2929, 2875, 1727, 1674, 1633, 1531, 1446, 1383,
1188, 1081 cmÀ1; HRMS (FAB): m/z calcd for C36H54N5O8 [M+H]+:
684.3972; found: 684.3984.
Compound 6n {2, 2, 2}: Yield (3 steps): 28% (2.0 mg, 2.99 mmol);
[a]2D8 =À171 (c=0.080, CHCl3); 1H NMR (600 MHz, CDCl3): d=7.76
(d, J=10.2 Hz, 1H), 7.37 (d, J=8.4 Hz, 1H), 7.21–7.30 (m, 5H), 5.29
(d, J=10.8 Hz, 1H), 5.06 (t, J=7.2 Hz, 1H), 4.94 (ddd, J=4.2, 8.4,
11.4 Hz, 1H), 4.64 (d, J=7.8 Hz, 1H), 4.47 (d, J=10.2 Hz, 1H), 4.12–
4.17 (m, 1H), 3.75 (t, J=9.0 Hz, 1H), 3.48–3.55 (m, 1H), 3.23 (brt,
J=12.6 Hz, 1H), 3.15 (s, 3H), 3.01 (s, 3H), 2.97–3.00 (m, 1H), 2.88–
2.96 (m, 2H), 2.84 (t, J=4.8 Hz, 1H), 2.74 (dd, J=5.4, 18.6 Hz, 1H),
2.59 (ddd, J=1.8, 12.0, 18.6 Hz, 1H), 2.58 (dd, J=3.0, 4.8 Hz, 1H),
2.38–2.45 (m, 1H), 2.30 (ddd, J=4.2, 7.2, 12.0 Hz, 1H), 2.17–2.22
(m, 1H), 2.12 (dd, J=6.6, 12.0 Hz, 1H), 2.00 (quin, J=7.2 Hz, 1H),
1.80–1.86 (m, 1H), 1.75–1.80 (m, 1H), 1.35–1.42 (m, 1H), 1.22–1.32
(m, 1H), 1.02–1.07 (m, 1H), 0.96 (d, J=6.0 Hz, 3H), 0.90 (t, J=
7.2 Hz, 3H), 0.87 (d, J=7.2 Hz, 3H), 0.84 ppm (d, J=6.6 Hz, 3H);
13C NMR (150 MHz, CDCl3): d=174.0, 172.8, 170.5, 170.3, 169.4,
168.1, 136.9, 128.9, 128.5, 127.0, 71.4, 65.7, 60.5, 55.8, 51.9, 47.9,
47.1, 46.5, 36.5, 34.2, 33.9, 33.5, 32.8, 29.8, 29.2, 28.7, 28.3, 24.7,
23.3, 19.7, 19.3, 16.5, 11.2 ppm; IR (neat): n˜ =2966, 2931, 2877,
1725, 1671, 1628, 1533, 1445, 1383, 1183, 1077 cmÀ1; HRMS (FAB):
m/z calcd for C35H52N5O8 [M+H]+: 670.3816; found: 670.3820.
Compound 6r {3, 3, 2}: Yield (3 steps): 34% (7.3 mg, 10.1 mmol);
1
[a]2D3 =À96.9 (c=0.235, CHCl3); H NMR (400 MHz, CDCl3): d=8.02
(d, J=9.2 Hz, 1H), 7.16–7.34 (m, 11 H), 5.55 (dd, J=4.8, 10.4 Hz,
1H), 5.19 (dt, J=4.0, 10.0 Hz, 1H), 4.99 (t, J=7.2 Hz, 1H), 4.77 (dd,
J=4.0, 10.8 Hz, 1H), 4.66 (d, J=7.6 Hz, 1H), 4.02–4.13 (m, 1H), 3.66
(t, J=9.2 Hz, 1H), 3.42–3.52 (m, 1H), 3.42 (t, J=11.6 Hz, 1H), 3.32
(s, 3H), 3.12 (brt, J=12.0 Hz, 1H), 3.03 (dd, J=4.0, 14.0 Hz, 1H),
2.96–3.03 (m, 1H), 2.88–2.93 (m, 1H), 2.83 (t, J=4.4 Hz, 1H), 2.79–
2.83 (m, 1H), 2.74 (s, 3H), 2.58–2.64 (m, 2H), 2.57 (dd, J=2.8,
4.4 Hz, 1H), 2.17–2.28 (m, 2H), 2.02 (dt, J=7.2, 14.4 Hz, 1H), 1.68–
1.82 (m, 3H), 1.17–1.25 (m, 1H), 1.07–1.25 (m, 1H), 0.67 (d, J=
6.8 Hz, 3H), 0.66 (d, J=6.4 Hz, 3H), À0.25 ppm (ddd, J=4.0, 8.4,
13.2 Hz, 1H); 13C NMR (150 MHz, CDCl3): d=173.6, 172.5, 170.2,
169.8, 168.8, 168.4, 136.5, 136.1, 129.4, 129.2, 128.9, 128.5, 127.2,
126.9, 70.7, 60.3, 58.4, 53.1, 50.9, 48.0, 47.0, 46.4, 37.9, 37.4, 36.5,
34.6, 33.4, 32.9, 30.9, 28.8, 28.6, 24.8, 23.1, 23.0, 22.0 ppm; IR (neat):
n˜ =2955, 2928, 2871, 1728, 1671, 1633, 1530, 1445, 1382, 1181,
Compound 6o {2, 3, 2}: Yield (3 steps): 28% (4.9 mg, 6.97 mmol);
[a]2D3 =À155 (c=0.140, CHCl3); 1H NMR (400 MHz, CDCl3): d=7.94
(d, J=9.6 Hz, 1H), 7.36 (d, J=9.2 Hz, 1H), 7.18–7.29 (m, 10H), 5.83
(t, J=7.6 Hz, 1H), 5.02–5.08 (m, 1H), 5.02 (t, J=6.8 Hz, 1H), 4.65 (d,
J=7.6 Hz, 1H), 4.44 (d, J=10.8 Hz, 1H), 4.10–4.18 (m, 1H), 3.69 (t,
J=9.6 Hz, 1H), 3.44–3.52 (m, 1H), 3.33 (dd, J=7.6, 14.4 Hz, 1H),
3.21 (s, 3H), 3.20 (brt, J=14.8 Hz, 1H), 3.08 (dd, J=8.0, 14.4 Hz,
1H), 2.97–3.04 (m, 2H), 2.88 (s, 3H), 2.84–2.88 (m, 1H), 2.84 (t, J=
4.8 Hz, 1H), 2.62–2.75 (m, 2H), 2.58 (dd, J=2.4, 4.8 Hz, 1H), 2.26–
2.30 (m, 1H), 2.22–2.26 (m, 1H), 2.15–2.22 (m, 1H), 2.02 (dt, J=6.8,
14.8 Hz, 1H), 1.71–1.80 (m, 2H), 1.12–1.21 (m, 1H), 0.85 (d, J=
6.4 Hz, 3H), 0.27 ppm (d, J=6.8 Hz, 3H); 13C NMR (150 MHz, CDCl3):
d=173.8, 172.7, 170.5, 169.7, 168.9, 168.4, 136.6, 136.1, 129.1,
129.0, 128.8, 128.5, 127.1, 126.9, 71.0, 66.0, 60.4, 51.7, 51.3, 47.9,
47.0, 46.4, 37.5, 35.7, 34.5, 33.4, 32.8, 30.5, 29.1, 28.5, 27.1, 23.2,
19.4, 17.8 ppm; IR (neat): n˜ =2964, 2927, 2879, 1726, 1671, 1630,
1532, 1446, 1384, 1222, 1182, 1077 cmÀ1; HRMS (FAB): m/z calcd for
C38H50N5O8 [M+H]+: 704.3659; found: 704.3648.
1078 cmÀ1
;
HRMS (FAB): m/z calcd for C39H52N5O8 [M+H]+:
718.3816; found: 718.3819.
General procedure III: Macrolactonization of 16
The synthesis of macrolactone 17 from 16 was performed in the
same manner as that used for the synthesis of 9; see general pro-
cedure I.
Compound 6p {3, 1, 2}: Yield (3 steps): 5% (1.6 mg, 2.32 mmol);
1
[a]2D5 =À186 (c=0.0450, CHCl3); H NMR (600 MHz, CDCl3): d=7.98
(d, J=9.0 Hz, 1H), 7.34 (d, J=9.0 Hz, 1H), 7.18–7.32 (m, 5H), 5.11–
5.16 (m, 1H), 5.05 (d, J=10.8 Hz, 1H), 5.04 (d, J=11.4 Hz, 1H), 5.02
(t, J=7.2 Hz, 1H), 4.65 (d, J=6.6 Hz, 1H), 4.13–4.18 (m, 1H), 3.68 (t,
J=9.6 Hz, 1H), 3.45–3.53 (m, 1H), 3.23 (s, 3H), 3.18 (brt, J=
12.0 Hz, 1H), 2.97–3.03 (m, 2H), 2.85 (s, 3H), 2.84 (t, J=4.2 Hz, 1H),
2.82–2.85 (m, 1H), 2.62–2.70 (m, 1H), 2.56–2.59 (m, 1H), 2.41–2.47
(m, 1H), 2.29–2.41 (m, 1H), 2.22–2.28 (m, 1H), 2.17–2.22 (m, 1H),
2.00–2.06 (m, 1H), 1.72–1.78 (m, 2H), 1.51–1.57 (m, 1H), 1.08–1.18
(m, 2H), 0.99 (d, J=6.0 Hz, 3H), 0.95 (d, J=5.4 Hz, 3H), 0.94 (d, J=
6.6 Hz, 3H), 0.90 ppm (d, J=6.6 Hz, 3H); 13C NMR (150 MHz, CDCl3):
d=173.7, 173.2, 170.4, 169.8, 168.9, 168.4, 136.7, 129.1, 128.5,
126.9, 70.9, 60.3, 58.4, 57.8, 51.2, 48.0, 47.0, 46.4, 39.3, 37.6, 34.5,
Compound 17a: Yield: 33% from polymer-supported H-bAla-OH
10 (8.2 mg, 11.6 mmol); [a]2D5 =À124 (c=0.388, CHCl3); 1H NMR
(400 MHz, CDCl3): d=8.32 (d, J=7.2 Hz, 1H), 7.06 (d, J=10 Hz, 1H),
5.21 (t, J=7.6 Hz, 1H), 5.03–5.08 (m, 2H), 4.93 (dd, J=6.0, 9.2 Hz,
1H), 4.65 (d, J=6.4 Hz, 1H), 3.95–4.15 (m, 3H), 3.91 (brt, J=8.8 Hz,
1H), 3.80 (q, J=8.8 Hz, 1H), 3.62 (t, J=6.8 Hz, 1H), 3.29 (t, J=
6.8 Hz, 2H), 3.23 (s, 3H), 3.04–3.15 (m, 1H), 2.69 (s, 3H), 2.52–2.67
(m, 2H), 2.45–2.50 (m, 1H), 2.12–2.18 (m, 2H), 2.00–2.10 (m, 1H),
1.94–1.98 (m, 2H), 1.80–1.91 (m, 2H), 1.71–1.80 (m, 2H), 1.58–1.68
(m, 2H), 1.42 (s, 3H), 1.32 (s, 3H), 1.31 (d, J=6.8 Hz, 3H), 1.25–1.30
(m, 4H), 0.89 (d, J=7.2 Hz, 3H), 0.85 ppm (d, J=7.6 Hz, 3H);
13C NMR (100 MHz, CDCl3): d=173.3, 173.0, 171.6, 170.8, 169.6,
Chem. Eur. J. 2015, 21, 18417 – 18430
www.chemeurj.org
18427
ꢀ 2015 Wiley-VCH Verlag GmbH & Co. KGaA, Weinheim

Full Paper
169.0, 109.2, 77.2, 71.6, 70.6, 69.1, 60.9, 55.6, 53.3, 52.3, 51.0, 46.5,
37.6, 34.6, 34.5, 33.5, 31.1, 29.5, 29.4, 28.8, 28.3, 26.9, 25.5, 24.3,
23.9, 15.6, 15.2, 11.3 ppm; IR (neat): n˜ =3383, 3302, 2958, 2929,
2875, 2097, 1732, 1665, 1640, 1519, 1446, 1408, 1379, 1345, 1303,
1258, 1231, 1180, 1102, 1061 cmÀ1; HRMS (FAB): m/z calcd for
C33H55N8O9 [M+H]+: 707.4092; found: 707.4073.
0.86 ppm (t, J=7.6 Hz, 3H); 13C NMR (100 MHz, CDCl3): d=173.2,
173.0, 171.5, 170.7, 169.6, 168.7, 77.2, 70.5, 60.9, 55.6, 53.4, 52.3,
51.0, 47.9, 47.1, 46.7, 38.1, 37.6, 34.5, 33.6, 33.4, 31.1, 28.8, 28.3,
25.7, 24.3, 23.9, 15.6, 15.2, 11.4 ppm; IR (neat): n˜ =3383, 3308,
2958, 2925, 2875, 2853, 2097, 1734, 1665, 1636, 1540, 1517, 1447,
1412, 1378, 1345, 1260, 1232, 1178, 1130, 1101, 1029 cmÀ1; HRMS
(FAB): m/z calcd for C30H49N8O8 [M+H]+: 649.3673; found:
649.3654.
Compound 17b: Yield: 31% from polymer-supported H-bAla-OH
10 (7.6 mg, 11.0 mmol); [a]2D6 =À221 (c=0.225, CHCl3); 1H NMR
(400 MHz, CDCl3): d=7.67 (d, J=10 Hz, 1H), 7.39 (d, J=8.8 Hz, 1H),
5.23 (q, J=6.8 Hz, 1H), 5.05 (t, J=6.8 Hz, 1H), 4.98 (d, J=10.8 Hz,
1H), 4.87 (dt, J=4.0, 9.6 Hz, 1H), 4.74 (d, J=7.2 Hz, 1H), 4.07–4.15
(m, 2H), 4.03 (m, 1H), 3.87 (t, J=10 Hz, 1H), 3.75 (q, J=10 Hz, 1H),
3.64 (t, J=9.6 Hz, 1H), 3.29 (dt, J=2.8, 10 Hz, 2H), 3.20 (s, 3H),
3.02–3.09 (m, 1H), 2.77 (s, 3H), 2.59–2.70 (m, 2H), 2.48–2.53 (m,
1H), 2.29–2.37 (m, 1H), 2.11–2.17 (m, 2H), 2.05–2.10 (m, 1H), 1.88–
1.97 (m, 2H), 1.62–1.73 (m, 1H), 1.54–1.59 (m, 2H), 1.46–1.54 (m,
2H), 1.42 (s, 3H), 1.33 (s, 3H), 1.31 (d, J=6.8 Hz, 3H), 0.92 (d, J=
6.4 Hz, 3H), 0.87 ppm (d, J=6.8 Hz, 3H); 13C NMR (100 MHz, CDCl3):
d=173.8, 173.5, 171.0, 170.7, 169.7, 168.8, 109.1, 77.2, 71.6, 70.8,
69.1, 60.5, 57.9, 55.3, 51.0, 49.7, 46.3, 34.5, 34.3, 33.2, 31.1, 30.4,
28.5, 28.2, 28.1, 27.4, 26.8, 25.5, 23.0, 19.8, 19.4, 15.0 ppm; IR (neat):
n˜ =3383, 3306, 2961, 2932, 2875, 2097, 1730, 1671, 1632, 1529,
1446, 1413, 1379, 1344, 1261, 1229, 1180, 1101, 1061 cmÀ1; HRMS
(FAB): m/z calcd for C32H52N8O9 [M+H]+: 693.3936; found:
693.3964.
Compound 15b: Yield (3 steps): 29% (2.0 mg, 3.15 mmol); [a]D27
=
1
À237 (c=0.100, CHCl3); H NMR (400 MHz, CDCl3): d=7.67 (d, J=
8.0 Hz, 1H), 7.40 (d, J=8.8 Hz, 1H), 5.25 (q, J=6.8 Hz, 1H), 4.99 (t,
J=6.8 Hz, 1H), 4.98 (d, J=10.4 Hz, 1H), 4.87 (dt, J=3.6, 10.0 Hz,
1H), 4.75 (d, J=7.6 Hz, 1H), 3.98–4.07 (m, 1H), 3.91 (brt, J=8.8 Hz,
1H), 3.57–3.68 (m, 1H), 3.30 (dt, J=2.8, 6.4 Hz, 2H), 3.20 (s, 3H),
3.06 (brt, J=11.6 Hz, 1H), 2.98–3.03 (m, 1H), 2.84 (t, J=4.4 Hz, 1H),
2.77 (s, 3H), 2.62–2.72 (m, 1H), 2.55–2.60 (m, 2H), 2.49–2.59 (m,
1H), 2.27–2.37 (m, 1H), 2.19–2.27 (m, 1H), 2.06–2.15 (m, 1H), 1.95–
2.06 (m, 1H), 1.88–1.95 (m, 2H), 1.53–1.73 (m, 4H), 1.44–1.52 (m,
2H), 1.31 (d, J=6.8 Hz, 3H), 0.92 (d, J=6.4 Hz, 3H), 0.87 ppm (d,
J=6.8 Hz, 3H); 13C NMR (100 MHz, CDCl3): d=173.8, 173.5, 170.9,
170.7, 169.7, 168.6, 70.7, 60.6, 57.9, 55.2, 51.0, 49.7, 47.9, 47.1, 46.5,
34.5, 33.4, 33.2, 31.1, 30.4, 28.4, 28.3, 28.1, 27.5, 24.1, 23.0, 19.8,
19.4, 15.1 ppm; IR (neat): n˜ =3384, 3305, 3062, 2961, 2925, 2875,
2849, 2097, 1729, 1669, 1631, 1528, 1446, 1414, 1379, 1344, 1260,
1228, 1178, 1119, 1100 cmÀ1; HRMS (FAB): m/z calcd for C29H47N8O8
[M+H]+: 635.3517; found: 635.3518.
Compound 17c: Yield: 20% from polymer-supported H-bAla-OH
10 (5.1 mg, 6.94 mmol); [a]2D5 =À142 (c=0.240, CHCl3); 1H NMR
(400 MHz, CDCl3): d=8.33 (d, J=9.2 Hz, 1H), 7.15 (d, J=9.2 Hz,
1H), 5.06 (dd, J=6.0, 8.0 Hz, 1H), 5.05 (d, J=11.2 Hz, 1H), 4.91 (dd,
J=6.4, 9.2 Hz, 1H), 4.80 (dd, J=2.8, 11.2 Hz, 1H), 4.70 (d, J=7.2 Hz,
1H), 4.04–4.13 (m, 3H), 3.90 (brt, J=9.6 Hz, 1H), 3.80 (m, 1H), 3.62
(dd, J=6.0, 7.2 Hz, 1H), 3.28 (t, J=6.8 Hz, 2H), 3.22 (s, 3H), 3.07–
3.13 (m, 1H), 2.75 (s, 3H), 2.61 (t, J=4.0 Hz, 2H), 2.47 (d, J=5.6 Hz,
1H), 2.27–2.37 (m, 4H), 2.09–2.17 (m, 2H), 2.03–2.08 (m, 1H), 1.92–
1.97 (m, 2H), 1.88–1.90 (m, 1H), 1.57–1.68 (m, 2H), 1.42 (s, 3H),
1.33 (s, 3H), 1.24–1.32 (m, 2H), 0.93 (d, J=7.2 Hz, 3H), 0.89 (d, J=
6.4 Hz, 3H), 0.85 (d, J=7.2 Hz, 3H), 0.84 ppm (t, J=7.2 Hz, 3H);
13C NMR (100 MHz, CDCl3): d=173.5, 173.4, 170.9, 170.8, 168.9,
168.4, 109.2, 77.2, 71.6, 70.9, 69.1, 60.8, 60.1, 57.7, 53.5, 51.1, 46.4,
37.5, 34.6, 34.5, 30.9, 30.2, 29.3, 29.0, 28.7, 27.3, 26.9, 25.5, 24.4,
23.9, 23.7, 20.0, 19.5, 15.4, 11.4 ppm; IR (neat): n˜ =3387, 3298,
2962, 2924, 2879, 2853, 2097, 1731, 1672, 1632, 1530, 1450, 1405,
1380, 1260, 1181 cmÀ1; HRMS (FAB): m/z calcd for C35H59N8O9 [M+
H]+: 735.4405; found: 735.4407.
Compound 15c: Yield (3 steps): 31% (1.5 mg, 2.16 mmol); [a]D25
=
1
À227 (c=0.0750, CHCl3); H NMR (400 MHz, CDCl3): d=8.34 (d, J=
8.0 Hz, 1H), 7.16 (d, J=9.2 Hz, 1H), 4.96–5.04 (m, 2H), 4.91 (dd, J=
6.8, 9.2 Hz, 1H), 4.81 (dd, J=2.4, 10.8 Hz, 1H), 4.71 (d, J=7.2 Hz,
1H), 4.06–4.16 (m, 1H), 3.95 (brt, J=8.8 Hz, 1H), 3.61–3.68 (m, 1H),
3.29 (t, J=6.8 Hz, 2H), 3.22 (s, 3H), 3.05–3.13 (m, 1H), 2.94–3.00 (m,
1H), 2.84 (t, J=4.8 Hz, 1H), 2.76 (s, 3H), 2.60–2.64 (m, 2H), 2.57
(dd, J=2.4, 4.8 Hz, 1H), 2.47–2.53 (m, 1H), 2.25–2.39 (m, 4H), 2.07–
2.10 (m, 1H), 1.92–2.03 (m, 4H), 1.88–1.92 (m, 1H), 1.57–1.68 (m,
2H), 1.32–1.38 (m, 2H), 1.27–1.31 (m, 2H), 0.94 (d, J=6.4 Hz, 3H),
0.89 (d, J=6.8 Hz, 3H), 0.85 (d, J=6.4 Hz, 3H), 0.84 ppm (t, J=
6.8 Hz, 3H); 13C NMR (100 MHz, CDCl3): d=173.4, 173.3, 170.8,
170.7, 168.6, 168.4, 77.2, 70.8, 60.9, 60.1, 57.7, 53.5, 51.1, 47.9, 47.1,
46.6, 37.5, 34.6, 33.6, 33.0, 30.9, 30.2, 29.0, 28.7, 27.3, 24.4, 24.0,
23.7, 20.0, 19.5, 15.4, 11.4 ppm; IR (neat): n˜ =3386, 3297, 2961,
2925, 2875, 2849, 2096, 1733, 1669, 1628, 1540, 1517, 1444, 1413,
1381, 1345, 1260, 1178, 1115, 1089, 1025 cmÀ1; HRMS (FAB): m/z
calcd for C32H53N8O8 [M+H]+: 677.3986; found: 677.3989.
General procedure IV: Synthesis of molecular probe 15
General procedure V: Preparation of mouse osteoclasts
from bone marrow cells
The synthesis of molecular probe 15 from 17 was performed in
the same manner as that used for the synthesis of 6; see general
procedure II.
For the preparation and culture of mouse osteoclasts, a-MEM sup-
plemented with 10% (v/v) heat-inactivated fetal calf serum (ICN Bi-
omedicals, Inc., Aurora, OH, USA), l-glutamine (4 mm), penicillin
(100 UmLÀ1), streptomycin (100 mgmLÀ1), and amphotericin B
(250 ngmLÀ1) was used in this study, and the cells were cultured at
378C in a humidified atmosphere of 5% (v/v) CO2 in air. Mouse os-
teoclasts were prepared from mouse bone marrow cells of six- to
nine-week-old male ddY mice (SLC Inc., Shizuoka, Japan). In brief,
mouse bone marrow cells (1.0–1.5”107 cells/dish/6 mL) were cul-
tured for the first 3 days in the presence of 4000 UmLÀ1 recombi-
nant human M-CSF (M-CSF), 1 ngmLÀ1 recombinant human TGF-b1
(TGF-b1), and 100 ngmLÀ1 GST-human sRANKL (GST-RANKL) in
100 mm diameter type I collagen-coated culture dishes (AGC
Compound 15a: Yield (3 steps): 24% (1.8 mg, 2.77 mmol); [a]D22
=
1
À172 (c=0.0900, CHCl3); H NMR (400 MHz, CDCl3): d=8.32 (d, J=
7.2 Hz, 1H), 7.06 (d, J=9.2 Hz, 1H), 5.20 (t, J=7.2 Hz, 1H), 5.08 (q,
J=6.8 Hz, 1H), 4.98 (t, J=7.2 Hz, 1H), 4.92 (dd, J=6.0, 9.2 Hz, 1H),
4.66 (d, J=7.2 Hz, 1H), 3.93–4.05 (m, 2H), 3.58–3.69 (m, 1H), 3.29
(t, J=6.8 Hz, 1H), 3.23 (s, 3H), 3.11 (brt, J=12.0 Hz, 1H), 2.92–2.97
(m, 1H), 2.83 (t, J=4.4 Hz, 1H), 2.70 (s, 3H), 2.63–2.69 (m, 1H),
2.50–2.58 (m, 2H), 2.29–2.43 (m, 1H), 2.23–2.31 (m, 1H), 1.99–2.10
(m, 1H), 1.92–1.99 (m, 3H), 1.83–1.92 (m, 2H), 1.55–1.68 (m, 2H),
1.30 (d, J=6.8 Hz, 3H) 1.23–1.37 (m, 4H), 0.87 (d, J=7.2 Hz, 3H),
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Rand, T. Rezai, C. M. McEwen, V. M. Gelev, R. A. Turner, R. G. Linington,
S. S. F. Leung, A. S. Kalgutkar, J. N. Bauman, Y. Zhang, S. Liras, D. A. Price,
A. M. Mathiowetz, M. P. Jacobson, R. S. Lokey, Nat. Chem. Biol. 2011, 7,
810–817; c) J. G. Beck, J. Chatterjee, B. Laufer, M. U. Kiran, A. O. Frank, S.
Neubauer, O. Ovadia, S. Greenberg, C. Gilon, A. Hoffman, H. Kesseler, J.
Am. Chem. Soc. 2012, 134, 12125–12133; d) F. Giordanetto, J. Kihlberg,
J. Med. Chem. 2014, 57, 278–295; e) A. Bhat, L. R. Roberts, J. J. Dwyer,
Eur. J. Med. Chem. 2015, 94, 471–479.
Techno Glass Co., Ltd., Chiba, Japan), in which GST-RANKL and Leu-
koprol were used for the preparation of mouse osteoclasts instead
of RANKL and M-CSF, respectively, according to previous re-
ports.[27–29] TGF-b1 was added to the culture medium together with
M-CSF and GST-RANKL, according to previous reports to promote
the efficiency of osteoclast differentiation.[30–33] After 3 days of cul-
ture, adherent cells were harvested by treatment with trypsin–eth-
ylenediaminetetraacetic acid (EDTA), and the resulting harvested
cells (3”104 cells/well/100 mL) were placed on 96-well half-area cul-
ture plates (Corning Inc., Corning, NY, USA) to promote the effi-
ciency of osteoclastic fusion and maturation. This study with exper-
imental animals was approved by and conducted in accordance
with the guidelines of the Animal Experiment Committee of Chubu
University.
[2] a) R. Furumai, A. Matsuyama, N. Kobashi, K. H. Lee, M. Nishiyama, H. Na-
kajima, A. Tanaka, Y. Komatsu, N. Nishino, M. Yoshida, S. Horinouchi,
Cancer Res. 2002, 62, 4916–4921; b) K. M. Vander Molen, W. McCulloch,
C. J. Pearce, N. H. Oberlies, J. Antibiot. 2011, 64, 525–531.
[3] a) A. Rügger, A. M. Kuhn, H. Lichti, H. R. Loosi, R. Huguenin, C. Quiquer-
ez, A. V. von Wartburg, Helv. Chim. Acta 1976, 59, 1075–1092; b) D.
Faulds, K. L. Goa, O. Benfield, Drugs 1993, 45, 953–1040.
[4] a) J. N. Steenbergen, J. Alder, G. M. Thorne, F. P. Tally, J. Antimicrob. Che-
mother. 2005, 55, 283–288; b) R. H. Baltz, V. Miao, S. K. Wrigley, Nat.
Prod. Rep. 2005, 22, 717–741.
[5] J. Chatterjee, C. Gilon, A. Hoffman, H. Kessler, Acc. Chem. Res. 2008, 41,
1331–1342.
General procedure VI: TRAP staining
Osteoclasts cultured with 0.5% (v/v) vehicle (DMSO) or test com-
pounds for 24 h were stained for tartrate-resistant acid phospha-
tase (TRAP), a typical marker enzyme of osteoclasts, as previously
described[13,34,35] to visualize osteoclasts. In brief, the cells were
fixed with 3.7% formaldehyde in phosphate-buffered saline (PBS)
for 15 min. After treatment with ethanol/acetone (1:1) for 1 min,
the well surface was dried and treated with the TRAP staining solu-
tion (0.1m sodium acetate buffer (pH 5.0) containing 50 mm
sodium tartrate, 0.1 mgmLÀ1 naphthol AS-MX phosphate, and
1 mgmLÀ1 fast red violet LB salt) for 30 min. The resulting visual-
ized osteoclasts were detected under an all-in-one fluorescence mi-
croscope (BZ-8100 BIOZERO; KEYENCE Corp., Osaka, Japan), with
which the number of osteoclasts with more than five nuclei were
also counted. Similar results were obtained in more than two other
experiments.
[6] a) M. Pa¨ıs, B. C. Das, P. Ferron, Phytochemistry 1981, 20, 715–723; b) S.
Gupta, D. W. Roberts, J. A. A. Renwick, J. Chem. Soc. Perkin Trans. 1 1989,
2347–2357.
[7] a) T. Kobayashi, S. Ikeno, N. Hosokawa, Y. Uehara, M. Hori, K. Tsuchiya,
Biol. Pharm. Bull. 2004, 27, 587–590; b) Y. Sasazawa, Y. Futamura, E. Ta-
shiro, M. Imoto, Cancer Sci. 2009, 100, 1460–1467; c) R. Dornetshuber-
Fleiss, P. Heffeter, T. Mohr, P. Hazemi, K. Kryeziu, C. Seger, W. Berger, R.
Lemmens-Gruber, Biochem. Pharmacol. 2013, 86, 361–377.
[8] F. Odier, A. Vey, J. Bureau, Biol. Cell 1992, 74, 267–271.
[9] S. Naganuma, N. Kuzuya, K. Sakai, K. Hasumi, A. Endo, Biochim. Biophys.
Acta 1992, 1126, 41–48.
[10] N. Itoh, M. Okochi, S. Tagami, K. Nishitomi, T. Nakayama, K. Yanagida, A.
Fukutomi, J. Jiang, K. Mori, M. Hosono, J. Kikuchi, Y. Nakano, Y. Takinami,
K. Dohi, A. Nishigaki, H. Takemoto, K. Minagawa, T. Katoh, M. Willem, C.
Haass, T. Morihara, T. Tanaka, T. Kudo, H. Hasegawa, M. Nishimura, G. Sa-
kaguchi, A. Kato, M. Takeda, Neurodegener. Dis. 2009, 6, 230–239.
[11] M. J. Vµzquez, M. I. Albarrµn, A. Espada, A. Rivera-Sagredo, E. Díaz, J. A.
Hueso-Rodríguez, Chem. Biodiversity 2005, 2, 123–130.
[12] S. R. Sennoune, K. Bakunts, G. M. Martínez, J. L. Chua-Tuan, Y. Kebir,
M. N. Attaya, R. Martínez-Zaquliµn, Am. J. Physiol. Cell. Physiol. 2004,
286, C1443–C1452.
[13] H. Nakagawa, M. Takami, N. Udagawa, Y. Sawae, K. Suda, T. Sasaki, N. Ta-
kahashi, M. Wachi, K. Nagai, J. T. Woo, Bone 2003, 33, 443–455.
[14] M. Yoshida, H. Takeuchi, Y. Ishida, M. Yashiroda, M. Yoshida, M. Takagi, K.
Shin-ya, T. Doi, Org. Lett. 2010, 12, 3792–3795.
[15] a) C. Farina, S. Gagliardi, Drug Discovery Today 1999, 4, 163–172; b) C.
Farina, S. Gagliardi, Curr. Pharm. Des. 2002, 8, 2033–2048; c) E. J.
Bowman, B. J. Bowman, J. Bioenerg. Biomembr. 2005, 37, 431–435;
d) F. L. Yuan, X. Li, W. G. Lu, C. W. Li, J. P. Li, Y. Wang, Mol. Biol. Rep. 2010,
37, 3561–3566; e) N. Katner, Y. Yao, G. J. Crasto, A. Datti, M. F. Manolson,
J. Biol. Chem. 2010, 285, 37476–37490; f) C. S. Thudium, V. K. Jensen,
M. A. Karsdal, K. Henriksen, Curr. Protein Pept. Sci. 2012, 13, 141–151.
[16] Y. Che, D. C. Swenson, J. B. Gloer, B. Koster, D. Malloch, J. Nat. Prod.
2001, 64, 555–558.
[17] a) I. Shiina, R. Ibuka, M. Kubota, Chem. Lett. 2002, 286–287; b) I. Shiina,
M. Kubota, R. Ibuka, Tetrahedron Lett. 2002, 43, 7535–7539; c) I. Shiina,
M. Kubota, H. Oshiumi, M. Hashizume, J. Org. Chem. 2004, 69, 1822–
1830; d) I. Shiina, M. Hashizume, Y. Yamai, H. Oshiumi, T. Shimazaki, Y.
Takasuna, R. Ibuka, Chem. Eur. J. 2005, 11, 6601–6608; e) I. Shiina, Bull.
Chem. Soc. Jpn. 2014, 87, 196–233.
General procedure VII: Assay for OCL morphology
Osteoclasts prepared from bone marrow cells were further cultured
in 96-well culture plates for 24 h after the culture medium was re-
placed with fresh medium that contained 4000 UmLÀ1 M-CSF,
100 ngmLÀ1 sRANKL, and 0.5% (v/v) vehicle (DMSO) or test com-
pounds. After culturing, cells were fixed and stained for TRAP and
then the number of osteoclasts with more than five nuclei with
a clear cytoplasm and smooth periphery were counted. Similar
results were obtained in more than two other experiments.
Acknowledgements
This work was supported by the JSPS through a Grant-in-Aid
for Scientific Research (B) (no. 26282208 for T.D.) and a Grant-
in-Aid for Young Scientists (B) (no. 23710264 for M.Y.), as well
as the Protein Research Foundation. This work was partially
supported by the Platform for Drug Discovery, Informatics, and
Structural Life Science from the Ministry of Education, Culture,
Sports, Science and Technology, Japan, and the Hori Sciences
and Arts Foundation.
[18] H. M. Geysen, R. H. Meloen, S. J. Barteling, Proc. Natl. Acad. Sci. USA
1984, 81, 3998–4002.
[19] SynPhase Lanterns were purchased from Mimotopes, 11 Duerdin Street,
Clayton, Victoria, 3168 Australia.
[20] PyBroP, see; E. FrØrot, J. Coste, A. Pantaloni, M. N. Dufour, P. Jouin, Tetra-
hedron 1991, 47, 259–270.
[21] Scalable synthesis of acid 7 was performed in the accordance with the
reported procedure, see: M. Yoshida, H. Sato, Y. Ishida, H. Nakagawa, T.
Doi, J. Org. Chem. 2014, 79, 296–306.
Keywords: combinatorial chemistry
peptides solid-phase synthesis
relationships
·
molecular probes
·
·
·
structure–activity
[1] a) T. Rezai, B. Yu, G. L. Millhauser, M. P. Jacobson, R. S. Lokey, J. Am.
Chem. Soc. 2006, 128, 2510–2511; b) T. R. White, C. M. Renzelman, A. C.
[22] J. D. Wade, J. Bedford, R. C. Sheppard, G. W. Tregear, Pep. Res. 1991, 4,
194–199.
Chem. Eur. J. 2015, 21, 18417 – 18430
www.chemeurj.org
18429
ꢀ 2015 Wiley-VCH Verlag GmbH & Co. KGaA, Weinheim

Full Paper
[23] The cyclization precursors 8 were immediately used for macrolactamiza-
tion without purification because decomposition of 8 was observed.
[24] a) R. Huisgen, Angew. Chem. Int. Ed. Engl. 1963, 2, 565–598; Angew.
Chem. 1963, 75, 604–637; b) H. C. Kolb, M. G. Finn, K. B. Sharpless,
Angew. Chem. Int. Ed. 2001, 40, 2004–2021; Angew. Chem. 2001, 113,
2056–2075; c) C. W. Tornøe, C. Christensen, M. Meldal, J. Org. Chem.
2002, 67, 3057–3064.
[25] Synthesis of Fmoc-l-N-Me-Lys(N3)-OH was performed according to the
reported procedure, see: T. Doi, Y. Numajiri, T. Takahashi, M. Takagi, K.
Shin-ya, Chem. Asian J. 2011, 6, 180–188.
[29] Y. Tomimori, K. Mori, M. Koide, Y. Nakamichi, T. Ninomiya, N. Udagawa,
H. Yasuda, J. Bone Miner. Res. 2009, 24, 1194–1205.
[30] R. J. Sells Galvin, C. L. Gatlin, J. W. Horn, T. R. Fuson, Biochem. Biophys.
Res. Commun. 1999, 265, 233–239.
[31] T. Kaneda, T. Nojima, M. Nakagawa, A. Ogasawara, H. Kaneko, T. Sato, H.
Mano, M. Kumegawa, Y. Hakeda, J. Immunol. 2000, 165, 4254–4263.
[32] K. Fuller, J. M. Lean, K. E. Bayley, M. R. Wani, T. J. Chambers, J. Cell Sci.
2000, 113, 2445–2453.
[33] T. Yan, B. L. Riggs, W. J. Boyle, S. Khosla, J. Cell. Biochem. 2001, 83, 320–
325.
[26] Synthesis of Fmoc-l-Lys(N3)-OH was performed according to the report-
ed procedure, see: A. Le Chevalier Isaad, F. Barbetti, P. Rovero, A. M.
D’Ursi, M. Chelli, M. Chorev, A. M. Papini, Eur. J. Org. Chem. 2008, 5308–
5314.
[34] H. Nakagawa, M. Wachi, J. T. Woo, M. Kato, S. Kasai, F. Takahashi, I. S.
Lee, K. Nagai, Biochem. Biophys. Res. Commun. 2002, 292, 94–101.
[35] H. Nakagawa, K. Hasumi, M. Takami, S. Aida-Hyugaji, J. T. Woo, K. Nagai,
T. Ishikawa, M. Wachi, Biochem. Pharmacol. 2007, 73, 34–43.
[27] A. Mochizuki, M. Takami, T. Kawawa, R. Suzumoto, M. Sasaki, A. Shiba, H.
Tsukasaki, B. Zhao, R. Yasuhara, T. Suzawa, Y. Miyamoto, Y. Choi, R.
Kamijo, J. Immunol. 2006, 177, 4360–4368.
Received: July 29, 2015
[28] B. Zhao, M. Takami, A. Yamada, X. Wang, T. Koga, X. Hu, T. Tamura, K.
Ozato, Y. Choi, L. B. Ivashkiv, H. Takayanagi, R. Kamijo, Nat. Med. 2009,
15, 1066–1071.
Revised: September 17, 2015
Published online on November 4, 2015
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