Design, synthesis and antifungal evaluation of borrelidin derivatives

Article abstract of DOI:10.1016/j.bmc.2018.11.005

Borrelidin, a nitrile containing 18-membered polyketide macrolide, display potent antifungal activity. In this study, a library of borrelidin derivatives were synthesized. Their structures were elucidated by detailed spectroscopic data analysis. The antifungal activity and cytotoxicity of these target compounds were evaluated by broth microdilution and 3-(4,5-dimethylthiazol-2-yl)-3,5-phenytetrazoliumromide (MTT) methods. Among forty-seven prepared analogues, compound 3b had the inhibitory effect on Candida albicans and Candida parapsilosis (MIC: 50 and 12.5 μg/mL, respectively). Furthermore, compounds 4n and 4r presented better antifungal activity against Aspergillus fumigatus with 12.5 μg/mL MIC value, which were insensitive to borrelidin. Preliminary structure-activity relationships (SAR) revealed that the ester analogues containing fragment -OCH₂CH₂N- had an important effect on the antifungal activity. Meanwhile, the molecular docking study indicated the carboxyl substituents in BN could provide extra interaction with pathogenic fungal threonyl-tRNA synthetase (ThrRS).

Full text of DOI:10.1016/j.bmc.2018.11.005

Accepted Manuscript
Design, Synthesis and Antifungal Evaluation of Borrelidin Derivatives
Caijuan Hu, Hao Su, Jinghan Luo, Li Han, Qingyin Liu, Wenxi Wu, Yu Mu,
Peipei Guan, Tiemin Sun, Xueshi Huang
PII:
S0968-0896(18)31430-5
https://doi.org/10.1016/j.bmc.2018.11.005
BMC 14609
DOI:
Reference:
To appear in:
Bioorganic & Medicinal Chemistry
Received Date:
Revised Date:
Accepted Date:
8 August 2018
24 October 2018
4 November 2018
Please cite this article as: Hu, C., Su, H., Luo, J., Han, L., Liu, Q., Wu, W., Mu, Y., Guan, P., Sun, T., Huang, X.,
Design, Synthesis and Antifungal Evaluation of Borrelidin Derivatives, Bioorganic & Medicinal Chemistry (2018),
doi: https://doi.org/10.1016/j.bmc.2018.11.005
This is a PDF file of an unedited manuscript that has been accepted for publication. As a service to our customers
we are providing this early version of the manuscript. The manuscript will undergo copyediting, typesetting, and
review of the resulting proof before it is published in its final form. Please note that during the production process
errors may be discovered which could affect the content, and all legal disclaimers that apply to the journal pertain.

Bioorganic & Medicinal Chemistry
Design, Synthesis and Antifungal Evaluation of Borrelidin Derivatives
a
a
b
a,
a
a
a
a
Caijuan Hu , Hao Su , Jinghan Luo , Li Han *, Qingyin Liu , Wenxi Wu , Yu Mu , Peipei Guan , Tiemin
b,
a,
Sun * and Xueshi Huang *
a
College of Life and Health Sciences, Northeastern University, Shenyang 110819, China
Key Laboratory of Structure-Based Drug Design and Discovery, Shenyang Pharmaceutical University, Shenyang 110016, China
b
ARTICLE INFO
ABSTRACT
Article history:
Received
Received in revised form
Accepted
Borrelidin, a nitrile containing 18-membered polyketide macrolide, display potent antifungal
activity. In this study, a library of borrelidin derivatives were synthesized. Their structures were
elucidated by detailed spectroscopic data analysis. The antifungal activity and cytotoxicity of
these target compounds were evaluated by broth microdilution and 3-(4,5-dimethylthiazol-2-yl)-
3,5-phenytetrazoliumromide (MTT) methods. Among forty-seven prepared analogues,
compound 3b had the inhibitory effect on Candida albicans and Candida parapsilosis (MIC: 50
and 12.5 μg/mL, respectively). Furthermore, compounds 4n and 4r presented better antifungal
activity against Aspergillus fumigatus with 12.5 μg/mL MIC value, which were insensitive to
borrelidin. Preliminary structure-activity relationships (SAR) revealed that the ester analogues
Available online
Keywords:
Borrelidin
Synthesis
Antifungal activity
Structure-activity relationships
Molecular docking
2 2
containing fragment -OCH CH N- had an important effect on the antifungal activity.
Meanwhile, the molecular docking study indicated the carboxyl substituents in BN could
provide extra interaction with pathogenic fungal threonyl-tRNA synthetase (ThrRS).
2009 Elsevier Ltd. All rights reserved.
1
. Introduction
a narrow therapeutic window due to its strong cytotoxic
activity. The multiple bioactivities of BN have been confirmed
2
5
Invasive candidiasis and aspergillosis serve as the mainly life-
to be correlated with BN suppressing the threonyl-tRNA
threatening fungal infections, especially in immunocompromised
patients. The majority of these invasive infections are mostly
caused by Candida albicans, Candida parapsilosis and
Aspergillus fumigatus. Unfortunately, limited few classes of
antifungal drugs such as polyenes and azoles, had various
drawbacks in terms of toxicity, spectrum of activity, and drug
resistance to increase the risk of treatment failure. Searching
for novel antifungal agents from natural products and their
26-29
formation in organisms.
The expression level or sequence
1
alteration of ThrRS from bacteria to human cells was associated
30
with the potency of inhibition of BN.
2,3
The derivatization of BN was limited because of its
complicated total synthesis and low output from fermentation of
strains. Therefore, few literatures had evaluated the structure-
activity relationships (SAR) of BN. In the process of our search
for antimicrobial active natural products from actinomycetes,
Streptomyces vinaceusdrappus (YIM 100880) with a high-yield
4
,5
6
derivatives is a significant approach. Borrelidin (BN, Fig. 1), an
1
Streptomyces roche in 1949. The gross structure of BN was
8-membered macrolide polyketide, was initially isolated from
7
31
borrelidin was isolated from the Hylobates hoolock feces. Our
firstly elucidated in 1967 and subsequently refined by detailed
preliminary research indicated that BN was active in vitro against
Candida parapsilosis (MIC, 50 μg/mL) and no activity against
Candida albicans and Aspergillus fumigatus. In an effort to
investigate the antifungal SAR of BN and discover promising
antifungal leads with low toxicity, a series of borrelidin
derivatives (BNs) were designed and synthesized in the present
research. Meanwhile, the docking studies were carried out to
explore the new BNs-binding pocket of fungal ThrRS.
8
,9
NMR analysis. Its absolute configuration was confirmed by X-
1
0
ray crystallography in 1989. Since then, the macrolide BN was
1
1-14
15-17
continuously focused on in total synthesis,
biosynthesis,
1
8-20
21-24
isolation,
and bioactivity,
which highlighted its
significance. Especially, BN presented strong inhibitory activity
against gram-positive, few gram-negative species of bacteria,
1
9,20
actinomyces and fungicidal activity.
———
Corresponding author.
Tel.: +86-24-83656122; E-mail: hanli@mail.neu.edu.cn (L.H.);
However, so far BN had
—


2
. Results and Discussion
2
.1. Chemistry
A detailed structural and functional analysis of the binding of
BN to ThrRS revealed that the plane of the 18-membered ring in
BN played a central role in binding the -sheet of the catalytic
3
0
domain of ThrRS. Notably, the carboxyl acid group and the
cyano moiety were closely correlation with the ThrRS-specific
recognition of BN in the different kingdom of life. Thus, a series
of BNs with the modifications of cyano and carboxyl groups
were designed and synthesized in the present research to discover
the promising leads against C. albicans, C. parapsilosis and A.
fumigatus. As showed in Fig. 1, the total design strategies of BNs
include: (a) hydrolyzation of nitrile functional group to obtain
compound 1; (b) amidation of the carboxyl group at C-23
position to synthesize borrelidin amide analogues 2a-2s; (c)
esterification of the carboxyl group at C-23 position to prepare
borrelidin ester analogues 3a-3i and 4a-4r.
Scheme 1. Preparation of compound 1. Reagents and
conditions: (i) CH CH=NOH, CuO, H O/MeOH (1:1), 100℃,
2 h.
3
2
1
Scheme 2. Preparation of compound 2a-2s. Reagents
and conditions: (ii) PyBoP, TEA, appropriate amines,
DCM, r.t, 6-24 h.
Figure 1. Total design of novel borrelidin derivatives.
On the basis of the above design strategy, the structure and
synthetic routes to these target compounds were described in
Schemes 1-4. These reaction conditions were relatively mild
without hydrolyzing macrocyclic lactone ring and the yields were
high. There was no predecessor that had hydrolyzed the cyano
group of BN by organic chemistry method. The hydration of
nitriles in BN could not be performed on the strong base
condition since lactone ring was instable for acids and bases. A
3
2
metal catalyst method was firstly used to yield compound 1 in
present study (Scheme 1). Nitrogen heterocyclic played an
important role in the numerous antifungal regeants to improve its
3
3-36
pharmacokinetic properties and biological activity.
Consequently, some suitable nitrogen heterocyclic were
introduced to structural skeleton via amidation and esterification.
Compounds 2a-2s, 3a-3i and 4a-4r were synthesized as depicted
in Schemes 2-4. Among them, compounds 2a-2s and 3a-3i were
prepared via a condensation reaction of BN and different amines
and alcohols in the presence of PyBOP or DCC, respectively.
Compound 4a was obtained by condensation of BN and 1-Boc-4-
(
2-hydroxyethyl)-piperazine and further to give 4b by the Boc
Scheme 3. Preparation of compound 3a-3i. Reagents and
3
deprotection. Compounds 4c-4r were synthesized from 4b by N-
conditions: (iii) DCC, HoBt, R OH (appropriate alcohols),
acylation or alkylation.
THF, r.t, 12-24 h.

inhibition effect against A. fumigatus with 12.5-50 μg/mL MICs.
Particularly, the MIC value of compound 4n and 4r against A.
fumigatus could unexpectedly reach 12.5 μg/mL, whereas BN
had no inhibition effect at a high concentration of 100 μg/mL.
a
Table 1. In vitro antifungal activity of some compounds
b
Comp
MIC (μg/mL)
c
d
e
C.a
C.p
A.f
25
50
50
_
f
_
3
3
3
3
3
3
4
4
4
4
a
b
c
d
e
f
50
12.5
25
100
100
100
100
100
50
_
_
100
100
_
_
n
o
p
r
12.5
25
_
_
_
_
_
_
_
Scheme 4. Preparation of compound 4a-4r. Reagents and conditions:
(
iv) (Boc)
2
O, TEA, DCM, r.t, 12 h; (v) TFA/DCM (V/V = 1:1),
50
4
0
℃→ r.t., 0.5 h; (vi) K
2
3
CO , R X (appropriate acyl halides or
halohydrocarbons), NaI, CH
3
CN, 80℃, 2 h.
12.5
2
.2. Evaluation of Antifungal Activity
_
Borrelidin
50
The antifungal activities of all synthetic compounds were
g
AmB
2.0
1.0
2.0
evaluated in vitro by a micro-dilution method to obtain their
minimum inhibitory concentration (MIC) values against three
pathogenic fungi, Candida albicans ATCC MYA-2876, Candida
parapsilosis ATCC 22019 and Aspergillus fumigatus CCTCC AF
a
The compounds (1, 2a-2s, 3g-3i, 4a-4m and 4q) had no inhibitory
effect on the tested pathogenic strains at the concentration 100
μg/mL.
9
3048. Amphotericin B was used as positive control. The MIC
b
MIC was defined as the minimal inhabitory concentration of a
values were summarized in Table 1.
compound. Identical values were obtained for each compound in three
replicates by visual investigation as stated in the experimental section.
The MIC values of compound 1 were all greater than 100
c
Candida albicans ATCC MYA-2876
μg/mL against the tested strains, which demonstrated compound
1
antifungal activity compared with BN. Combined with the
with converting the cyano group to primary amide had lost
d
Candida parapsilosis ATCC 22019
e
2
0
Aspergillus fumigatus CCTCC AF 93048
activity result and previous literature, it further confirmed that
the cyanide group on BN was necessary for its antifungal
activity. All of borrelidin amide analogues (2a-2s), which were
synthesized by introducing some nitrogen heterocyclic and amino
acid compounds, were found to be no antifungal activity at the
concentration of 100 μg/mL. Amide analogues were possibly
unable to recognize the specific residues of the enzymatic site of
fungal ThrRS, resulting in the resistance of BNs binding to
ThrRS.
f
_
The “ ” symbol represents no effect at the highest tested
concentration (100 μg/mL )
g
Amphotericin B (AmB) was used as positive control.
2
.3. Structure-Activity Relationship Summary
In combination with the analysis of antifungal activity,
the primary structure-activity relationship was summarised
in Fig. 2. The loss of cyano group led to the significant
decrease of activity (1 vs. BN). The antifungal activity of all
amide analogues (2a-2s) were dramatically decreased or
disappeared. The most of ester analogues (3a-3i, 4a-4r)
were apparently better activities than BN, which indicated
that the modification of the carboxyl group to the ester-
type analogues could produce novel antifungal agents with
new structural scaffolds. As can be seen from Fig. 2, the
fragment (-OCH CH N-) was quite favorable for BNs to
In order to further explore the effects of the modifications of
carboxyl group in BN on the antifungal activity, we attempted to
construct the derivatives (3a-3i) through ester bond in the
following research. Besides, piperazine moieties as an important
component of many antifungal agents were also implanted into
BNs (4a-4r). The results showed that analogues 3b-3f displayed
the inhibition effects against C. albicans and C. parapsilosis with
MIC values in a range of 12.5-100 μg/mL. More significantly,
ester analogues 3b-3f presented some inhibition effect against C.
albicans, while BN was inactive at 100 μg/mL concentration. In
comparison with BN (MIC: 50 μg/mL), only ester analogues 3b
3
7
2
2
exert antifungal activity. At the same time, the strong
electron donating ability of nitrogen in the fragment (-
OCH CH N-) is beneficial for improving antifungal activity
(MIC: 12.5 μg/mL) and 3c (MIC: 25 μg/mL) had excellent
2
2
antifungal activity against C. parapsilosis. Among all of BNs,
compouds 3a-3d, 4n-4p, and 4r showed a certain extent
(3b-3f vs. 3a). Although the introduction of imidazole

group was unfavorable for inhibiting C. albicans and C.
parapsilosis, it was benefit to improve the activity against A.
fumigatus. Unsubstituted piperazine ring had not change
antimicrobial spectrum of borrelidin (4b vs. BN), but N-
alkyl substituted piperazine ring introduced into BN favored
to improve its activity against A. fumigatus compared with
N-acyl/aromatic substituted piperazine ring (3c, 4n-4r vs.
Table 2. IC50 (μg/mL) and selectivity index (SI) values of active
compounds against 293T
a
b
Comp
IC50 (μg/mL)
SI value
C.a
4
b-4m). Only N-methyl piperazine derivative of BNs (3c)
c
presented the activity against C. albicans and C.
parapsilosis.
293T
C.p
A.f
d
3
3
3
a
b
c
18.09±0.42
35.18±0.56
58.46±0.31
27.62±0.28
39.23±0.26
28.02±0.74
<0.18
<0.18
2.81
0.72
0.71
1.17
0.55
<0.39
<0.28
>5.61
0.92
0.34
1.76
<0.14
2
.4. Cytotoxicity and selectivity index of BNs
0.71
To create BNs’ selectivity between fungi and human
38
cells, in vitro cytotoxicities of BNs (3a-3f, 4n-4p and 4r)
were further examined using human kidney epithelial cell
line (293T) by MTT method. As described in Table 2, the
cytotoxicities of all the tested derivatives dropped obviously
in comparation with BN. The selectivity index value of
derivative 3b with the best activity against C. albicans, was
at least five times that of BN. Compounds 3b and 3c with
satisfying activity against C. parapsilosis gave good
selectivity index values (SI: 2.81, 2.34). Remarkably, The
SI values of derivatives (3c, 4n and 4r) possessing the
excellent activity against A. fumigatus were 8-40-fold
superior to that of BN. Overall, these results suggested that
borrelidin analogs (3b, 3c, 4n and 4r) have a potential
usefulness in development of drugs for infectious diseases
and pathogenic fungi control.
0.58
2.34
3d
3e
3f
0.28
0.28
0.39
0.39
0.28
0.28
e
4
4
4
4
n
o
p
r
>70.15
<0.70
<0.23
<0.17
<0.22
<0.14
<0.70
<0.23
<0.17
<0.22
0.28
23.02±1.00
17.04±0.62
22.04±0.70
13.97±0.15
Borrelidin
a
IC50 is defined as the concentration at which 50% growth is
inhibited.
b
Selectivity index (in vitro) = cytotoxicity (IC50: μg/mL)/antifungal
activity (MIC: μg/mL)
c
2
93T: human kidney epithelial cell line
d
The MIC values of these compounds > 100 μg/mL.
The IC50 value of 4n > 100 μM
e.
Figure 2. Structure-activity relationships of borrelidin

Figure 3. Molecular binding mode of BN (A1) and 3b (A2) into the active pocket of ThrRS (protein surface diagram in software
PyMOL); the bingding modes of BN (B1) and 3b (B2) with the key residues of ThrRS (3D interaction relationship diagram in software
Discovery Studio 3.0; Purple: hydrophobic interaction, Green: hydrogen bond, Red: ionic bond); hydrogen bonding interactions of BN
(
C1) and 3b (C2) with the key residues of ThrRS (2D interactions in Glide module; Purple: hydrogen bond, The arrow point to the
hydrogen bond acceptor).
2
.5. Molecular Docking
Molecular docking is an effective and reliable tool able to
3
. Conclusions
In summary, a series of antifungal borrelidin derivatives were
locate the probable binding interactions of ligands with their
target proteins.
synthetase (ThrRS). Due to the absence of crystal structure of
ThrRS from C. albicans, C. parapsilosis and A. fumigatus in the
Protein Data Bank, the docking studies were performed based on
the X-ray structure of the yeast mitochondrial ThrRS (PDB ID:
designed and synthesized via the rational design. The SAR
indicated that the presence of nitrile group and the fragment (-
OCH CH N-) introduced into carboxyl of BN were essential for
antifungal activity. The biological assay displayed a good
selectivity index between fungi and human cells for the active
BNs (3b, 4n and 4r), which were greatly promising antifungal
candidates. Furthermore, the molecular docking investigation
indicated the carboxyl substituents in BN existed extra
interaction with ThrRS of pathogenic fungi, which provided
useful guidelines for new tRNA synthetase inhibitor.
3
9,40
BN is a natural inhibitor of threonyl-tRNA
23
2
2
4
yye). Hence, BN and the potent analogue 3b were docked in the
active site of ThrRS to predict binding mode and support the
antifungal results (Fig. 3). Although the compound 3b filled the
similar binding space as BN, the side chain of 3b was projected
deeper into the bottom of binding pocket than BN as shown in
figure 3A1/A2. Compared to BN, 3b did not establish hydrogen
bonds with Asn108, Glu164 and Arg162 through 8/16-hydroxyl
groups but form hydrophobic interactions with Pro232 and
Met131 through 9/15-methyl groups (Fig. 3B1/B2). The great
distinction of 3b and BN depended on the formation of hydrogen
bonds between 23-position substituted group and different amino
acids in ThrRS. Gln287 and Lys273 oriented toward 23-position
carbonyl of 3b to form two hydrogen bonds. Oxygen atom in the
fragment -OCH CH N- of 3b was positioned toward amino group
4. Experimental Section
4
.1. General Experimental Procedures
The synthetic routes to borrelidin derivatives 1, 2a-2s,
a-3i and 4a-4r were depicted in Schemes 1-4. All the
3
target compounds were fully analyzed and characterized by
1
13
H and C NMR spectra, high resolution mass spectrometry
(
ESI-HRMS), optical rotations, ultraviolet spectra and IR
2
2
spectra before biological screening. Column
of guanidyl on Arg162 to make hydrogen bond. Both methylene
group in the fragment -OCH CH N- and the pyrrole-NH of 3b
were also involved in the interactions with active site residue
Gln292. Moreover, the binding mode revealed that the compound
3
chromatography were Silica gel (100-200 mesh, 200-300
mesh, Qingdao Marine Chemical Ltd., Qingdao, China),
Sephadex LH-20 (GE Healthcare Biosciences AB, Uppsala,
Sweden), YMC*GEL ODS-A (S-50 μm, 12 nm) (YMC Co.,
Ltd., Kyoto, Japan), and Amberlite XAD-16 polymeric resin
2
2
b bearing the pyrrolidine ring created a strong ionic bond via
interacting with the important acidic amino acid Asp182.
Perhaps, this was the primary cause of its improving antifungal
activity. According to the in vitro antifungal activity and
molecular docking results, it was established that the 23-position
substituted group of BNs extended into a new orthogonal pocket,
which underwent induced-fit nature of BNs’ interaction with
ThrRS of pathogenic fungi.
(
Rohm and Hass ShanghaiChemical Industry Co., Ltd.,
Shanghai, China) were used for columnchromatography.
MTT and antimicrobial assays were analyzed using
amicroplate reader (Bio Tek Synergy H1, Bio Tek
Instruments, Inc., Vermont, USA).
4
.1.1. Synthesis of (1R,2R)-2-
[(2S,4E,6E,8R,9S,11R,13S,15S, 16S)-7-carbamoyl-8-
hydroxy-9,11,13,15,16-pentamethyl-18-
oxooxacyclooctadeca-4,6-dien-2-yl]cyclopentane-1-
carboxylic acid (1)
To a 10 mL round-bottom flask were added BN 20 mg
0.041 mmol), acetaldoxime 4.84 mg (0.082 mmol), copper
(
oxide 1.59 mg (0.020 mmol) and H O/MeOH (v/v =1:1, 3
2
mL). The mixture was heated to reflux for 12 h. The
completion of reaction was checked with monitoring TLC.
After cooling to room temperature, the reaction mixture
was directly evaporated to dryness and the residue was
purified by column chromatography on silica gel
(
dichloromethane/methanol = 25:1) to give 1 as a light-
20
yellow crystal (70% yield): []
D
-3.4 (c 1.18, MeOH); UV
(MeOH) λmax (log ε) 256 (4.33) nm; IR v 3409, 2952,

2
9
1
925, 1718, 1658, 1578, 1454, 1384, 1267, 1182, 1023,
(600 MHz, CD OD) δ (ppm): 6.93 (d, J = 11.3 Hz, 1H),
3
-1
1
76 cm ; H NMR (600 MHz, CD OD) δ (ppm): 6.91 (d, J =
6.60 (dd, J = 14.9, 11.3 Hz, 1H), 6.39 (ddd, J = 14.9, 8.6,
6.6 Hz, 1H), 4.98 (dt, J = 9.9, 3.4 Hz, 1H), 4.18 (d, J = 9.8
Hz, 1H), 3.90 (ddd, J = 8.0, 4.5, 3.2 Hz, 1H), 3.70-3.59 (m,
5H, 2 × -CH -N-CO-), 3.54-3.47 (m, 3H, 2 × -CH O-), 2.97-
3
1.4 Hz, 1H), 6.46 (dd, J = 14.9, 11.4 Hz, 1H), 6.13 (ddd,
J = 14.9, 10.5, 4.6 Hz, 1H), 4.85 (dt, J = 10.5, 3.7 Hz, 1H),
4
2
.21 (d, J = 10.0 Hz, 1H), 3.80 (dt, J = 10.3, 2.9 Hz, 1H),
.59 (quint, J = 9.4 Hz, 1H), 2.49 (ddd, J = 14.9, 10.6, 4.2
2
2
2.90 (m, 2H), 2.57 (m, 2H), 2.31 (dd, J = 11.3, 3.6 Hz, 1H),
2.27 (d, J = 11.3 Hz, 1H), 2.12-2.00 (m, 2H), 1.86-1.69 (m,
5H), 1.67 (m, 1H), 1.56 (m, 1H), 1.47 (m, 1H), 1.24-1.19
(m, 2H), 1.11 (brt, J = 12.5 Hz, 1H), 1.04 (d, J = 6.4 Hz,
3H, CH ), 0.98-0.93 (m, 2H), 0.87 (d, J = 6.9 Hz, 3H, CH ),
Hz, 1H), 2.42 (brd, J = 14.9 Hz, 1H), 2.32 (q, J = 7.7 Hz,
1
1
H), 2.28 (dd, J = 16.0, 2.9 Hz, 1H), 2.12 (dd, J = 16.0,
0.1 Hz, 1H), 1.92-1.86 (m, 2H), 1.84 (m, 1H), 1.78-1.62
(m, 5H), 1.50 (m, 1H), 1.31 (m, 1H), 1.06 (m, 2H), 0.94 (d,
J = 6.3 Hz, 3H, CH ), 0.93 (m, 1H), 0.87 (m, 1H), 0.84 (m,
3
3
0.85 (d, J = 6.4 Hz, 3H, CH ), 0.84 (d, J = 6.0 Hz, 3H, CH ),
3
3
3
1
3
1
H), 0.75 (d, J = 6.7 Hz, 3H, CH ), 0.74 (d, J = 6.7 Hz, 3H,
0.70 (brt, J = 12.5 Hz, 1H); C NMR (150 MHz, CD OD) δ
3
3
CH ), 0.67 (d, J = 6.5 Hz, 3H, CH ), 0.58 (brt, J = 13.2 Hz,
(ppm): 174.9, 171.5, 144.0, 139.0, 127.1, 118.4, 115.9,
76.5, 71.6, 70.4, 66.4, 66.3, 47.8, 45.9, 45.2, 44.2, 43.1,
42.3, 38.3, 37.5, 35.6, 35.6, 34.4, 31.0, 29.0, 27.0, 26.0,
24.3, 19.3, 17.0, 16.9, 13.9. ESI-HRMS (m/z): calcd. for
3
3
13
1
1
4
2
H); C NMR (150 MHz, CD OD) δ (ppm): 174.9, 172.5,
71.8, 136.7, 136.3, 132.9, 128.0, 76.2, 73.2, 71.6, 48.9,
8.2, 46.1, 43.4, 38.3, 36.9, 35.6, 35.2, 34.4, 31.1, 29.2,
3
+
7.1, 26.3, 24.9, 19.6, 17.6, 17.2, 14.6. ESI-HRMS (m/z):
C H N O [M+H] : 559.3747; found, 559.3749.
32
51
2
6
+
calcd. for C H NO [M+H] : 508.3274; found, 508.3273.
2
8
46
7
4
.1.2.3. (2S,4E,6Z,8R,9S,11R,13S,15S,16S)-8,16-
dihydroxy-9,11, 13,15-tetramethyl-2-[(1R,2R)-2-
piperidine-1-carbonyl)cyclo-pentyl]-18-
4
.1.2. General Procedure for the Synthesis of Compounds 2a-2s
(
To a stirred solution of BN (30 mg, 0.061 mmol) in DCM
oxooxacyclooctadeca-4,6-diene-7-carbonitrile (2c)
(
1.0 mL) was added TEA (12.4 mg, 0.122 mmol) and
PyBoP (35.1 mg, 0.067 mmol) at room temperature. After
0 min, the appropriate amine (0.153 mmol) was added.
Eluent petroleum ether/EtOAc (3:1). Colorless crystal.
2
0
3
89% yield: []
D
-4.4 (c 0.92, MeOH); UV (MeOH) λmax
Keep stirring for 6-24 h, the reaction mixture was washed
with 3% HCl aqueous (2 × 10 mL), saturated NaHCO3
aqueous (2 × 10 mL) and brine (2 × 10 mL). The resulting
organic layer was dried over Na SO , filtered and
(log ε) 254 (4.56) nm; IR v 3428, 2927, 2864, 2212, 1725,
1618, 1454, 1378, 1253, 1187, 1126, 1036, 975, 854, 812,
-1
1
755, 696 cm ; H NMR (600 MHz, CD OD) δ (ppm): 6.94
3
(d, J = 11.3 Hz, 1H), 6.62 (brt, J = 11.3 Hz, 1H), 6.40 (m,
1H), 5.00 (dt, J = 10.6, 3.5 Hz, 1H), 4.20 (d, J = 9.8 Hz,
1H), 3.92 (dt, J = 9.8, 3.3 Hz, 1H), 3.57 (m, 2H, 1 × -CH2-
2
4
concentrated in vacuo. The residue was purified by silica
gel column chromatography eluting with appropriate
mixture as indicated in each case. The data of 2k was
N-CO-), 3.49 (m, 2H, 1 × -CH -N-CO-), 2.95 (m, 2H), 2.57
2
4
1
found in previous literature.
(m, 2H), 2.27 (m, 2H), 2.04 (m, 2H), 1.79 (m, 5H), 1.68
m, 2H), 1.55 (m, 6H, 3 × -CH -), 1.45 (m, 1H), 1.23 (m,
2
4
1
.1.2.1. (2S,4E,6Z,8R,9S,11R,13S,15S,16S)-8,16-dihydroxy-9,11,
3,15-tetramethyl-2-[(1R,2R)-2-(piperazine-1-carbonyl)cyclo-
H), 1.11 (m, 1H), 1.05 (d, J = 6.4 Hz, 3H, CH ), 0.97 (m,
3
13
H), 0.86 (brd, J = 6.4 Hz, 9H, 3 × CH ), 0.70 (m, 1H);
C
3
pentyl]-18-oxooxacyclooctadeca-4,6-diene-7-carbonitrile (2a)
3
Eluent dichloromethane/methanol (40:1). light-yellow
139.2, 127.2, 118.5, 115.9, 76.6, 71.6, 70.6, 48.0, 46.5,
45.4, 44.3, 43.2, 43.1, 37.9, 37.5, 35.7, 35.6, 34.5, 31.2,
2
0
crystal. 96% yield: []
D
-16 (c 0.82, MeOH); UV (MeOH)
λmax (log ε) 254 (4.50) nm; IR v 3399, 2956, 2921, 2873,
211, 1720, 1631, 1446, 1374, 1262, 1180, 1037, 974 cm ;
29.2, 26.9, 26.4, 26.1, 25.5, 24.4, 24.1, 19.4, 17.1, 17.0,
-1
+
2
13.9. ESI-HRMS (m/z): calcd. for C33
579.3774; found, 579.3770.
H N
52
O
2
5
Na [M+Na] :
1
H NMR (600 MHz, CD OD) δ (ppm): 6.93 (d, J = 11.3 Hz,
3
1
1
H), 6.60 (dd, J = 14.9, 11.3 Hz, 1H), 6.39 (ddd, J = 14.9,
0.0, 5.3 Hz, 1H), 4.98 (dt, J = 9.8, 3.6 Hz, 1H), 4.19 (d, J
9.8 Hz, 1H), 3.90 (m, 1H), 3.68-3.54 (m, 4H, 2 × -CH2-
4
.1.2.4. (2S,4E,6Z,8R,9S,11R,13S,15S,16S)-8,16-
dihydroxy-9,11, 13,15-tetramethyl-2-[(1R,2R)-2-
pyrrolidine-1-carbonyl)cyclo-pentyl]-18-
=
(
N-CO-), 3.00-2.85 (m, 6H, 2 × -CH -N-, 2 × -CH-), 2.60-
2
oxooxacyclooctadeca-4,6-diene-7-carbonitrile (2d)
2
(
1
.53 (m, 2H), 2.28 (m, 2H), 2.12-2.00 (m, 2H), 1.86-1.69
m, 5H), 1.67 (m, 1H), 1.56 (m, 1H), 1.47 (m, 1H), 1.26-
.18 (m, 2H), 1.11 (brt, J = 12.6 Hz, 1H), 1.04 (d, J = 6.4
Eluent dichloromethane/methanol (80:1). Colorless
2
0
crystal. 83% yield: []
-16.9 (c 1.06, MeOH); UV (MeOH)
λmax (log ε) 252 (4.68) nm; IR v 3400, 2954, 2878, 2212,
D
Hz, 3H, CH ), 0.98-0.93 (m, 2H), 0.87 (d, J = 6.9 Hz, 3H,
3
CH ), 0.85 (d, J = 6.4 Hz, 3H, CH ), 0.84 (d, J = 6.0 Hz, 3H,
1723, 1619, 1452, 1380, 1260, 1180, 1087, 1040, 976, 832,
3
3
13
-1
1
CH ), 0.70 (brt, J = 12.6 Hz, 1H); C NMR (150 MHz,
788, 737, 657 cm ; H NMR (600 MHz, CD OD) δ (ppm):
3
3
CD OD) δ (ppm): 174.7, 171.5, 144.0, 139.0, 127.1, 118.4,
6.90 (d, J = 11.3 Hz, 1H), 6.60 (dd, J = 14.9, 11.3 Hz, 1H),
6.35 (m, 1H), 5.01 (dt, J = 10.6, 3.5 Hz, 1H), 4.17 (d, J =
9.8 Hz, 1H), 3.88 (dt, J = 9.2, 3.3 Hz, 1H), 3.45 (t, J = 6.8
Hz, 2H, 1 × -CH -N-CO-), 3.38 (m, 2H, 1 × -CH -N-CO-),
3
1
4
2
15.9, 76.5, 71.6, 70.2, 45.3, 45.0, 44.7, 44.3, 44.2, 43.1,
1.3, 38.7, 37.4, 35.7, 35.6, 34.4, 30.9, 29.0, 27.0, 26.0,
4.2, 19.3, 17.0, 16.7, 13.8. ESI-HRMS (m/z): calcd. for
2
2
+
C H N O [M+H] : 560.3907; found, 560.3910.
2.82 (m, 1H), 2.72 (q, J = 9.0 Hz, 1H), 2.54 (dd, J = 7.5,
32
52
3
5
3
1
×
1
1
.1 Hz, 2H), 2.23 (dd, J = 16.1, 9.5 Hz, 1H), 2.19 (dd, J =
6.1, 3.7 Hz, 1H), 2.11-2.00 (m, 2H), 1.98-1.83 (m, 3H, 2
4
.1.2.2. (2S,4E,6Z,8R,9S,11R,13S,15S,16S)-8,16-
dihydroxy-9,11, 13,15-tetramethyl-2-[(1R,2R)-2-
morpholine-4-carbonyl)cyclo-pentyl]-18-
-CH -), 1.84-1.74 (m, 6H), 1.65 (m, 1H), 1.59 (m, 1H),
2
(
.44 (m, 1H), 1.20 (m, 2H), 1.09(brt, J = 12.6 Hz, 1H),
oxooxacyclooctadeca-4,6-diene-7-carbonitrile (2b)
.03 (d, J = 6.4 Hz, 3H, CH ), 0.93-0.98 (m, 2H), 0.84 (d, J
3
Eluent dichloromethane/methanol (40:1). light-yellow
= 6.2 Hz, 6H, CH
3
), 0.83 (d, J = 6.8 Hz, 3H, CH
3
), 0.67 (brt,
J = 12.6 Hz, 1H); C NMR (150 MHz, CD OD) δ (ppm):
3
13
2
0
crystal. 96% yield: []
D
-16 (c 0.82, MeOH); UV (MeOH)
λmax (log ε) 254 (4.50) nm; IR v 3488, 3430, 2957, 2921,
175.0, 171.5, 144.1, 139.0, 127.3, 118.5, 116.0, 76.5, 71.6,
70.9, 48.0, 46.5, 45.8, 44.7,43.2, 37.6, 37.3, 35.8, 35.6,
34.5, 30.5, 28.9, 27.0, 26.1, 25.5, 24.2, 24.0, 19.5, 17.4,
2
1
864, 2212, 1723, 1628, 1525, 1456, 1379, 1265, 1229,
-1 1
182, 1116, 1075, 1036, 976, 857, 790, 700 cm ; H NMR

7.1, 13.9. ESI-HRMS (m/z): calcd. for C H N O Na
3
2
51
2
5
+
Eluent dichloromethane/methanol (80:1). light-yellow
2
0
D
-4.1 (c 0.97, MeOH); UV (MeOH)
213, 1721, 1628, 1460, 1384, 1261, 1163, 1087, 1035,
-
1
1
44, 731, 656 cm ; H NMR (600 MHz, CD OD) δ (ppm):
3
.95 (d, J = 11.3 Hz, 1H), 6.63 (dd, J = 14.9, 11.3 Hz, 1H),
.41 (ddd, J = 14.9, 8.4, 6.2 Hz, 1H), 5.00 (dt, J = 10.3,
.6 Hz, 1H), 4.20 (d, J = 9.8 Hz, 1H), 3.92 (m, 1H), 3.76-
.46 (m, 4H, 2 × -CH -N-CO-), 2.95 (m, 2H), 2.64-2.46 (m,
2
H), 2.39 (s, 3H, -N-CH ), 2.31 (dd, J = 11.6, 4.0 Hz, 1H),
3
.27 (d, J = 11.6 Hz, 1H), 2.15-2.00 (m, 2H), 1.89-1.72 (m,
H), 1.68 (m, 1H), 1.57 (m, 1H), 1.48 (m, 1H), 1.23 (brt, J
11.9 Hz, 2H), 1.12 (brt, J = 11.6 Hz, 1H), 1.06 (d, J =
.5 Hz, 3H, CH ), 0.98-0.94 (m, 2H), 0.88 (d, J = 6.6 Hz,
3
H, CH ), 0.86 (d, J = 6.5 Hz, 3H, CH ), 0.85 (d, J = 6.5 Hz,
3
3
13
H, CH ), 0.70 (brt, J = 11.6 Hz, 1H); C NMR (150 MHz,
3
3
1
4
2
15.9, 76.6, 71.7, 70.5, 54.5, 54.1, 47.9, 47.5, 45.4, 44.6,
4.3, 43.2, 41.2, 38.4, 37.5, 35.8, 35.7, 34.5, 31.1, 29.1,
1723, 1624, 1456, 1377, 1265, 1138, 1041, 978, 848, 742,
-1
1
645 cm ; H NMR (600 MHz, CD OD) δ (ppm): 6.82 (d, J =
3
7.0, 26.1, 24.3, 19.4, 17.2, 17.0, 13.9. ESI-HRMS (m/z):
11.3 Hz, 1H), 6.50 (dd, J = 14.9, 11.3 Hz, 1H), 6.28 (ddd,
J = 14.9, 9.2, 6.0 Hz, 1H), 4.88 (dt, J = 10.0, 3.6 Hz, 1H),
+
calcd. for C H N O [M+H] : 572.4063; found, 572.4063.
3
3
54
3
5
.08 (d, J = 9.8 Hz, 1H), 3.80 (dt, J = 8.6, 3.8 Hz, 1H),
4
(
.1.2.6. (2S,4E,6Z,8R,9S,11R,13S,15S,16S)-2-[(1R,2R)-2-
4-benzylpiperazine-1-carbonyl)cyclopentyl]-8,16-
dihydroxy-9,11,13,15-tetramethyl-18-oxooxacyclooctadeca-
.58 (t, J = 5.9 Hz, 2H, - CH -OH), 3.55 (m, 1H), 3.50-3.40
2
2
× -N-CH -), 2.38 (m, 2H), 2.19 (dd, J = 15.9, 5.6 Hz, 1H),
2
4
,6-diene-7-carbonitrile (2f)
.16 (d, J = 15.9 Hz, 1H), 1.94 (m, 2H), 1.73-1.63 (m, 5H),
1.56 (m, 1H), 1.44 (m, 1H), 1.35 (m, 1H), 1.11 (m, 2H),
1.00 (brt, J = 12.5 Hz, 1H), 0.94 (d, J = 6.4 Hz, 3H, CH
0.88-0.82 (m, 2H), 0.76 (d, J = 6.4 Hz, 3H, CH ), 0.75 (d, J
= 6.4 Hz, 3H, CH ), 0.74 (d, J = 6.2 Hz, 3H, CH ), 0.59 (brt,
J = 12.5 Hz, 1H); C NMR (150 MHz, CD OD) δ (ppm):
3
2
0
),
3
3
2
1
3
3
13
NMR (600 MHz, CD OD) δ (ppm): 7.35 (m, 4H, Ar-H), 7.31
174.7, 171.6, 144.1, 139.1, 127.2, 118.5, 116.0, 76.5, 71.7,
70.6, 59.7, 58.3, 53.4, 52.9, 47.9, 45.4, 45.0, 44.3, 43.1,
41.5, 38.2, 37.5, 35.8, 35.7, 34.5, 31.1, 29.1, 27.0, 26.1,
3
(
m, 1H, Ar-H), 6.94 (d, J = 11.3 Hz, 1H), 6.62 (dd, J =
1
1
6
4.9, 11.3 Hz, 1H), 6.40 (m, 1H), 4.99 (brd, J = 9.5 Hz,
H), 4.20 (d, J = 9.8 Hz, 1H), 3.92 (m, 1H), 3.73-3.49 (m,
H, 2 × -CH -N-CO-, -N-CH -Ar), 2.94 (m, 2H), 2.58-2.48
24.4, 19.4, 17.2, 17.1, 13.9. ESI-HRMS (m/z): calcd. for
+
C
34
H N
56
O
3
[M+H] : 602.4169; found, 602.4169.
2
2
6
m, 6H), 2.28 (m, 2H), 2.05 (m, 2H), 1.85-1.75 (m, 5H),
.68 (m, 1H), 1.56 (m, 1H), 1.47 (m, 1H), 1.23 (brt, J =
2.3 Hz, 2H), 1.12 (brt, J = 11.7 Hz, 1H), 1.06 (d, J = 6.4
4
.1.2.9. (1R,2R)-2-[(2S,4E,6Z,8R,9S,11R,13S,15S,16S)-7-
cyano-8,16-dihydroxy-9,11,13,15-tetramethyl-18-
oxooxacyclooctadeca-4,6-dien-2-yl]-N'-
propionylcyclopentane-1-carbohydrazide (2i)
3
3
3
13
H); C NMR (150 MHz, CD OD) δ (ppm): 174.8, 171.6,
Eluent dichloromethane/methanol (100:1). Colorless
3
2
0
44.1, 139.2, 129.2 (2C), 128.1 (3C), 127.3, 127.2, 118.5,
15.9, 76.5, 71.6, 70.5, 52.8, 52.4, 48.0, 45.4, 44.9, 44.3,
3.2, 41.5, 38.3, 37.5, 35.8, 35.7, 34.5, 31.0, 29.1, 27.0,
crystal. 60% yield: []
λmax (log ε) 256 (4.46) nm; IR v 3431, 3375, 3258, 3034,
2957, 2924, 2213, 1714, 1619, 1463, 1377, 1262, 1209,
1181, 1107, 1038, 974, 814, 779, 626 cm ; H NMR (600
D
-8 (c 1.00, MeOH); UV (MeOH)
-1
1
6.1, 24.3, 19.4, 17.3, 17.0, 13.9. ESI-HRMS (m/z): calcd.
+
MHz, CD OD) δ (ppm): 6.89 (d, J = 11.3 Hz, 1H), 6.58 (dd,
3
9
58
3
5
3
J = 14.9, 11.3 Hz, 1H), 6.31 (ddd, J = 14.9, 10.8, 4.4 Hz,
4
.1.2.7. (2S,4E,6Z,8R,9S,11R,13S,15S,16S)-8,16-
1
1
2
2
H), 4.95 (dt, J = 10.6, 3.5 Hz, 1H), 4.17 (d, J = 9.8 Hz,
H), 3.90 (dt, J = 10.1, 3.1 Hz, 1H), 2.77 (m, 1H), 2.62-
.52 (m, 2H), 2.45 (brq, J = 7.0 Hz, 1H), 2.30-2.22 (m,
dihydroxy-2-[(1R,2R)-2-(4-(4-methoxyphenyl)piperazine-1-
carbonyl)cyclo-pentyl]-9,11,13,15-tetramethyl-18-
oxooxacyclooctadeca-4,6-diene-7-carbonitrile (2g)
H), 2.23 (q, J = 7.2 Hz, 2H, NH-CO-CH -), 2.06-1.96 (m,
2
2H), 1.87-1.71 (m, 6H), 1.65 (m, 1H), 1.43 (m, 1H), 1.26-
1.19 (m, 2H), 1.14 (t, J = 7.2 Hz, 3H, CH ), 1.08 (brt, J =
12.2 Hz, 1H), 1.03 (d, J = 6.0 Hz, 3H, CH ), 1.00-0.93 (m,
2H), 0.86 (d, J = 6.0 Hz, 3H, CH ), 0.85 (d, J = 6.0 Hz, 6H,
2 × CH ), 0.68 (brt, J = 12.2 Hz, 1H); C NMR (150 MHz,
CD OD) δ (ppm): 176.0, 173.6, 172.1, 144.1, 138.9, 127.3,
118.5, 116.0, 76.2, 71.6, 71.5, 48.0, 47.2, 44.4, 43.2, 37.6,
2
0
8
(
1
8
6
3
3
3
13
3
3

3
1
7.4, 35.8, 35.3, 34.5, 31.9, 29.7, 27.1, 26.5, 26.2, 25.2,
(log ε) 254 (4.84) nm; IR v 3642, 3477, 3410, 3357, 2924,
9.4, 17.5, 17.2, 13.9, 8.6. ESI-HRMS (m/z): calcd. For
2211, 1724, 1647, 1543, 1447, 1372, 1259, 1180, 1104,
+
-1
1
C H N O [M+H] : 560.3700; found, 560.3700.
1038, 975, 850, 769, 699, 638 cm ; H NMR (600 MHz,
31
50
3
6
3
.1.2.10. (1R,2R)-N-butyl-2-
Eluent dichloromethane/methanol (80:1). Colorless
CH
2.96 (m, 2H, -CH
2
10.5, 4.1 Hz, 1H), 2.52 (brd, J = 15.1 Hz, 1H), 2.34 (dd, J
= 15.7, 3.0 Hz, 1H), 2.33 (t, J = 6.0 Hz, 1H), 2.25 (dd, J =
15.7, 10.0 Hz, 1H), 1.99 (m, 1H), 1.90 (m, 1H), 1.85-1.75
(m, 4H), 1.72 (m, 1H), 1.68-1.60 (m, 2H), 1.41 (m, 1H),
1.21 (brt, J = 13.0 Hz, 1H), 1.16 (brt, J = 13.0 Hz, 1H),
b
-NH-CO), 3.34 (dt, J = 13.4, 7.3 Hz, 1H, -CH
b
-NH-CO),
2
0
crystal. 85% yield: []
D
-8 (c 1.00, MeOH); UV (MeOH)
λmax (log ε) 256 (4.42) nm; IR v 3424, 2959, 2882, 2214,
-Py), 2.68 (m, 1H), 2.58 (ddd, J = 15.1,
1
6
1
4
(
×
2
2
3
1
718, 1623, 1455, 1376, 1272, 1201, 1121, 1037, 970, 689,
-1
1
51 cm ; H NMR (600 MHz, CD OD) δ (ppm): 6.93 (d, J =
3
1.3 Hz, 1H), 6.62 (brt, J = 12.3 Hz, 1H), 6.37 (m, 1H),
.98 (brd, J = 10.5 Hz, 1H), 4.19 (d, J = 9.8 Hz, 1H), 3.95
m, 1H), 3.44 (m, 1H), 3.35 (m, 1H), 3.25-3.14 (m, 2H, 1
-CH -N-CO-), 2.94 (m, 1H), 2.74 (brq, J = 8.3 Hz, 1H),
1.07 (brt, J = 12.4 Hz, 1H), 1.03 (d, J = 6.4 Hz, 3H, CH
0.95 (brt, J = 12.6 Hz, 1H), 0.91 (brt, J = 12.6 Hz, 1H),
3
),
2
.61 (brt, J = 14.5 Hz, 1H), 2.55 (brd, J = 14.5 Hz, 1H),
.24 (m, 2H), 2.09-2.05 (m, 2H), 1.91 (m, 2H), 1.81 (m,
H), 1.66 (m, 1H), 1.61 (m, 1H), 1.53 (m, 4H, 2 × -CH -),
0.84 (d, J = 6.2 Hz, 3H, CH
0.81 (d, J = 7.0 Hz, 3H, CH
3
), 0.82 (d, J = 6.4 Hz, 3H, CH
),
3
), 0.67 (brt, J = 12.4 Hz, 1H);
3
13
C NMR (150 MHz, CD OD) δ (ppm): 177.1, 171.5, 158.7,
2
3
.45 (m, 1H), 1.34 (m, 4H, 2 × -CH -), 1.26 (brt, J = 12.8
148.4, 144.0, 138.9, 137.2, 127.2, 123.6, 121.7, 118.4,
115.9, 76.2, 71.5, 71.0, 50.0, 47.8, 44.5, 43.1, 39.0, 37.6,
37.2, 35.6, 35.3, 34.4, 31.7, 37.0, 29.1, 27.0, 26.1, 24.6,
2
Hz, 1H), 1.17 (brt, J = 12.6 Hz, 1H), 1.11 (brt, J = 12.6 Hz,
H), 1.05 (d, J = 6.2 Hz, 3H, CH ), 1.02-0.92 (m, 2H), 0.97
1
3
(
(
(
(
7
3
2
t, J = 7.2 Hz, 3H, CH ), 0.96 (t, J = 7.2 Hz, 3H, CH ), 0.85
19.4, 17.4, 17.2, 13.9. ESI-HRMS (m/z): calcd. for
3
3
+
d, J = 6.4 Hz, 3H), 0.84 (d, J = 6.4 Hz, 6H, 2 × CH ), 0.68
C
35
H N
52
O
3
5
[M+H] : 594.3907; found, 594.3906.
3
13
brt, J = 12.6 Hz, 1H); C NMR (150 MHz, CD OD) δ
3
4
[
.1.2.13. methyl{(1R,2R)-2-
(2S,4E,6Z,8R,9S,11R,13S,15S,16S)-7-cyano-8,16-
dihydroxy-9,11,13,15-tetramethyl-18-oxooxacyclo-
ppm): 176.0, 171.6, 144.2, 139.0, 127.4, 118.5, 116.0,
6.7, 71.6, 71.0, 48.1, 47.7, 46.4, 45.8, 44.4, 43.1, 37.6,
6.4, 35.5, 35.4, 34.5, 31.8, 31.5, 29.9, 29.4, 26.8, 26.1,
octadeca-4,6-dien-2-yl]cyclopentane-1-carbonyl}-D-phenyl-
alaninate (2m)
4.6, 19.9, 19.7, 19.5, 17.8, 17.2, 14.0, 13.0, 12.9. ESI-
+
HRMS (m/z): calcd. For C H N O [M+H] : 623.4400;
3
6
61
2
5
found, 623.4401.
Eluent petroleum ether/EtOAc (3:1). Colorless crystal.
2
0
6
(
1
1
0% yield: []
D
-10.1 (c 0.79, MeOH); UV (MeOH) λmax
4
.1.2.11. (1R,2R)-N-benzyl-2-
log ε) 256 (4.55) nm; IR v 3421, 3366, 2956, 2923, 2213,
[(2S,4E,6Z,8R,9S,11R,13S,15S,16S) -7-cyano-8,16-
734, 1653, 1531, 1457, 1374, 1257, 1213, 1177, 1125,
-1 1
dihydroxy-9,11,13,15-tetramethyl-18-oxooxa-
cyclooctadeca-4,6-dien-2-yl]cyclopentane-1-carboxamide
087, 1035, 976, 738, 701, 643 cm ; H NMR (600 MHz,
CD OD) δ (ppm): 7.28 (t, J = 7.2 Hz, 2H, Ar-H), 7.22 (t, J
3
(2k)
=
7.2 Hz, 1H, Ar-H), 7.20 (d, J = 7.2 Hz, 2H, Ar-H), 6.88 (d,
1
H NMR (600 MHz, CD OD) δ (ppm): 7.31 (t, J = 7.3 Hz,
J = 11.3 Hz, 1H), 6.59 (dd, J = 14.9, 11.3 Hz, 1H), 6.26
(ddd, J = 14.9, 10.5, 4.6 Hz, 1H), 4.94 (dt, J = 10.3, 3.6
Hz, 1H), 4.56 (dd, J = 8.0, 6.8 Hz, 1H), 4.17 (d, J = 9.8 Hz,
3
2
6
6
3
1
1
2
H), 7.27 (d, J = 7.3 Hz, 2H), 7.24 (t, J = 7.3 Hz, 1H),
.90 (d, J = 11.3 Hz, 1H), 6.61 (dd, J = 14.9, 11.3 Hz, 1H),
.34 (ddd, J = 15.0, 10.7, 4.5 Hz, 1H), 4.98 (dt, J = 10.7,
.6 Hz, 1H), 4.45 (d, J = 15.1 Hz, 1H), 4.25 (d, J= 15.1 Hz,
H), 4.19 (d, J = 9.8 Hz, 1H), 3.86 (dt, J = 10.2, 3.0 Hz,
H), 2.77 (m, 1H), 2.60 (ddd, J = 15.0, 10.7, 4.1 Hz, 1H),
.54 (brd, J = 15.0 Hz, 1H), 2.45 (q, J = 7.9 Hz, 1H), 2.25
1H), 3.89 (dt, J = 10.1, 3.1 Hz, 1H), 3.63 (s, 3H, -OCH ),
3
3.09 (dd, J = 13.8, 6.8 Hz, 1H), 3.01 (dd, J = 13.8, 8.0 Hz,
1H), 2.64 (m, 1H), 2.56-2.46 (m, 3H), 2.37 (dd, J = 16.3,
2.9 Hz, 1H), 2.21 (dd, J = 16.3, 10.1 Hz, 1H), 2.00-1.88 (m,
2H), 1.85-1.76 (m, 3H), 1.76-1.68 (m, 3H), 1.63 (m, 1H),
1.41 (m, 1H), 1.22 (m, 1H), 1.18 (m, 1H), 1.06 (brt, J =
(
dd, J = 16.2, 2.8 Hz, 1H), 2.18 (dd, J = 16.2, 10.2 Hz,
H), 2.01 (m, 2H), 1.84-1.74 (m, 6H), 1.66 (m, 1H), 1.44
m, 1H), 1.24 (brt, J = 13.1 Hz, 1H), 1.08 (m, 2H), 1.04 (d,
1
(
12.5 Hz, 1H), 1.03 (d, J = 6.2 Hz, 3H, CH
), 0.93 (m, 2H),
3
0.83 (d, J = 6.8 Hz, 3H, CH ), 0.82 (d, J = 7.2 Hz, 6H, 2 ×
3
13
J = 6.4 Hz, 3H, CH ), 0.93 (m, 2H), 0.93 (m, 2H), 0.86 (d,
J = 6.6 Hz, 3H, CH ), 0.84 (d, J = 6.6 Hz, 3H), 0.77 (d, J =
CH
CD
3
), 0.70 (brt, J = 12.5 Hz, 1H); C NMR (150 MHz,
OD) δ (ppm): 177.2, 172.1, 171.7, 144.2, 139.1, 136.8,
3
3
3
13
7
.0 Hz, 3H, CH ), 0.71 (brt, J = 13.1 Hz, 1H); C NMR
128.8 (2C), 128.2 (2C), 127.3, 126.6, 118.5, 116.0, 75.9,
71.6, 71.1, 54.2, 51.2, 48.8, 47.9, 44.4, 43.2, 37.6, 37.4,
37.1, 35.7, 35.2, 34.6, 31.4, 29.6, 27.1, 26.2, 25.0, 19.4,
3
(150 MHz, CD OD) δ (ppm): 177.2, 171.6, 144.0, 138.9,
3
1
7
3
1
38.4, 128.1 (2C), 127.3, 127.0 (2C), 126.8, 118.4, 116.0,
6.2, 71.5, 71.3, 49.9, 47.9, 44.6, 43.1, 42.8, 37.6, 36.9,
5.5, 35.3, 34.5, 31.8, 29.4, 27.0, 26.1, 24.9, 19.4, 17.6,
17.4, 17.3, 14.0. ESI-HRMS (m/z): calcd. for C38
[M+Na] : 651.4009; found, 651.4012.
H
N
54
O
2
Na
7
+
+
7.1, 13.9. ESI-MS (m/z): 579.2000 [M+H] , 601.1000
+
4.1.2.14. methyl{(1R,2R)-2-
(2S,4E,6Z,8R,9S,11R,13S,15S,16S)-7-cyano-8,16-
[M+Na] , calcd. for C H N O .
3
5
50
2
5
[
4
.1.2.12. (1R,2R)-2-[(2S,4E,6Z,8R,9S,11R,13S,15S,16S)-7-
dihydroxy-9,11,13,15-tetramethyl-18-oxooxacyclo-
octadeca-4,6-dien-2-yl]cyclopentane-1-carbonyl}-D-
tryptophan-ate (2n)
cyano-8,16-dihydroxy-9,11,13,15-tetramethyl-18-
oxooxacyclooctadeca-4,6-dien-2-yl]-N-(2-(pyridin-2-
yl)ethyl)cyclopentane-1-carbox-amide (2l)
Eluent dichloromethane/methanol (100:1). White crystal.
2
0
Eluent petroleum ether/EtOAc (1:2). Colorless crystal.
63% yield: []
D
-7.4 (c 1.09, MeOH); UV (MeOH) λmax
2
0
8
8% yield: []
D
-18.7 (c 1.07, MeOH); UV (MeOH) λmax
(log ε) 220 (4.69), 256 (4.63) nm; IR v 3397, 2956, 2922,

2
1
212, 1731, 1646, 1524, 1459, 1379, 1353, 1256, 1213,
179, 1100, 1033, 976, 844, 746, 665 cm ; H NMR (600
4.8 Hz, 1H), 4.96 (dt, J = 10.5, 3.6 Hz, 1H), 4.37 (dd, J =
9.0, 6.0 Hz, 1H), 4.17 (d, J = 9.8 Hz, 1H), 3.93 (dt, J
-1
1
MHz, CD OD) δ (ppm): 7.50 (d, J = 7.9 Hz, 1H, Ar-H), 7.34
=10.2, 2.8 Hz, 1H), 3.69 (s, 3H, -OCH ), 2.74 (m, 1H),
3
3
(
7
d, J = 7.9 Hz, 1H, Ar-H), 7.09 (t, J = 7.9 Hz, 1H, Ar-H),
.08 (s, 1H, Ar-H), 7.01 (t, J = 7.9 Hz, 1H, Ar-H), 6.80 (d,
J = 11.3 Hz, 1H), 6.54 (dd, J = 14.9, 11.3 Hz, 1H), 6.15
ddd, J = 14.9, 10.5, 4.4 Hz, 1H), 4.93 (dt, J = 10.4, 3.6
Hz, 1H), 4.66 (t, J = 6.9 Hz, 1H), 4.15 (d, J = 9.8 Hz, 1H),
.83 (dt, J = 9.9, 3.1 Hz, 1H), 3.59 (s, 3H, -OCH ), 3.26
2.60-2.50 (m, 3H), 2.36 (dd, J = 16.1, 2.9 Hz, 1H), 2.24
(dd, J = 16.1, 10.3 Hz, 1H), 2.00-1.95 (m, 2H), 1.91-
1.70(m, 6H), 1.67 (m, 1H), 1.62-1.56 (m, 2H), 1.46 (m,
1H), 1.41 (m, 1H), 1.20-1.17 (m, 2H), 1.09 (brt, J = 12.5
Hz, 1H), 1.00-0.91 (m, 2H), 0.67 (brt, J = 12.5 Hz, 1H),
1.03 (d, J = 6.4 Hz, 3H, CH ), 0.96 (d, J = 6.6 Hz, 3H, CH ),
(
3
3
3
3
(
dd, J = 14.6, 6.9 Hz, 1H), 3.19 (overlapped, 1H), 2.66 (m,
0.93 (d, J = 6.6 Hz, 3H, CH ), 0.85 (d, J = 6.5 Hz, 3H, CH ),
3
3
1
2
1
1
1
1
6
H), 2.55-2.45 (m, 3H), 2.30 (dd, J = 16.3, 3.0 Hz, 1H),
.17 (dd, J = 16.3, 9.9 Hz, 1H), 1.99-1.90 (m, 2H), 1.82-
.67 (m, 6H), 1.62 (m, 1H), 1.40 (m, 1H), 1.19 (m, 1H),
.14 (m, 1H), 1.05 (brt, J = 12.6 Hz, 1H), 0.93 (brt, J =
3.0 Hz, 1H), 0.87 (brt, J = 13.0 Hz, 1H), 0.67 (brt, J =
0.84 (d, J = 7.1 Hz, 3H, CH ), 0.83 (d, J = 7.0 Hz, 3H, CH );
3 3
13
C NMR (150 MHz, CD OD) δ (ppm): 177.7, 173.3, 171.7,
3
144.1, 138.9, 127.3, 118.5, 116.0, 76.1, 71.6, 71.3, 51.2,
51.1, 48.6, 47.9, 43.9, 43.2, 40.0, 37.6, 36.9, 35.7, 35.3,
34.6, 31.7, 29.9, 27.1, 26.2, 25.2, 24.7, 21.9, 21.0, 19.5,
17.7, 17.2, 13.9. ESI-HRMS (m/z): calcd. for C H N O
2.6 Hz, 1H), 1.02 (d, J = 6.4 Hz, 3H, CH ), 0.81 (d, J =
3
35 57
2
7
13
+
.1 Hz, 6H, 2 × CH ), 0.71 (d, J = 6.8 Hz, 3H, CH );
C
[M+H] : 617.4166; found, 617.4162.
3
3
NMR (150 MHz, CD OD) δ (ppm): 177.1, 172.6, 171.8,
3
.1.2.17. methyl{(1R,2R)-2-
1
1
5
3
44.2, 139.0, 136.6, 127.3, 127.2, 123.0, 121.2, 118.5,
18.6, 117.8, 115.9, 111.0, 109.2, 76.0, 71.6, 71.1, 53.8,
1.2, 49.0, 47.9, 44.3, 43.0, 37.6, 37.3, 35.7, 35.2, 34.5,
1.3, 29.6, 27.3, 27.1, 26.3, 25.0, 19.4, 17.3, 17.3, 14.0.
+
ESI-HRMS (m/z): calcd. for C H N O [M+H] : 690.4118;
40
56
3
7
found, 690.4118.
Eluent dichloromethane/methanol (40:1). Colorless
20
crystal. 79% yield: []
D
-33.3 (c 0.24, MeOH); UV (MeOH)
4
.1.2.15. methyl{(1R,2R)-2-
λmax (log ε) 254 (4.83) nm; IR v 3323, 2956, 2924, 2854,
[(2S,4E,6Z,8R,9S,11R,13S,15S,16S)-7-cyano-8,16-
2
1
212, 1730, 1651, 1535, 1461, 1377, 1261, 1209, 1176,
dihydroxy-9,11,13,15-tetramethyl-18-oxooxacyclo-
octadeca-4,6-dien-2-yl]cyclopentane-1-carbonyl}-D-
methionin-ate (2o)
-1 1
093, 1041, 1024, 973, 802, 684, 650 cm ; H NMR (600
MHz, CD OD) δ (ppm): 7.60 (s, 1H, Ar-H), 6.90 (d, J =
3
1
1.1 Hz, 1H), 6.88 (s, 1H, Ar-H), 6.58 (dd, J = 14.9, 11.1
Hz, 1H), 6.30 (ddd, J = 14.9, 10.5, 4.8 Hz, 1H), 4.95 (dt, J
= 10.0, 3.7 Hz, 1H), 4.57 (t, J = 4.6 Hz, 1H), 4.17 (d, J =
9.7 Hz, 1H), 3.86 (dt, J = 9.8, 3.2 Hz, 1H), 3.65 (s, 3H, -
2
0
2
1
6
6
3
9
851, 2211, 1728, 1649, 1538, 1437, 1378, 1261, 1208,
OCH ), 3.05 (m, 2H), 2.70 (m, 1H), 2.59-2.48 (m, 3H),
3
-1
174, 1022, 972 cm ; 1H NMR (600 MHz, CD OD) δ (ppm):
2.33 (dd, J = 16.1, 3.2 Hz, 1H), 2.20 (dd, J = 16.1, 10.1
Hz, 1H), 2.00-1.95 (m, 2H), 1.84-1.68 (m, 6H), 1.63 (m,
1H), 1.42 (m, 1H), 1.20 (brt, J = 12.3 Hz, 2H), 1.08 (brt, J
= 11.4 Hz, 1H), 1.00-0.88 (m, 2H), 0.69 (brt, J = 11.4 Hz,
3
.90 (d, J = 11.3 Hz, 1H), 6.60 (dd, J = 14.9, 11.3 Hz, 1H),
.32 (ddd, J = 14.9, 10.6, 4.8 Hz, 1H), 4.96 (dt, J = 10.7,
.6 Hz, 1H), 4.49 (dd, J = 8.7, 5.2 Hz, 1H), 4.17 (d, J =
.8 Hz, 1H), 3.93 (dt, J = 10.4, 2.8 Hz, 1H), 3.71 (s, 3H, -
1H), 1.03 (d, J = 6.7 Hz, 3H, CH ), 0.83 (d, J = 6.6 Hz, 6H,
3
13
OCH ), 2.71 (m, 1H), 2.61-2.49 (m, 5H), 2.39 (dd, J = 16.3,
2 × CH ), 0.79 (d, J = 6.7 Hz, 3H, CH ); C NMR (150 MHz,
3
3
3
2
2
1
1
.7 Hz, 1H), 2.26 (dd, J = 16.3, 10.4 Hz, 1H), 2.10 (m, 1H),
CD OD) δ (ppm): 177.3, 171.9, 171.7, 144.1, 139.1, 135.0,
3
.08 (s, 3H, -SCH ), 2.03-1.95 (m, 2H), 1.94 (m, 1H), 1.89-
127.2, 118.6, 117.2, 115.9, 76.0, 71.6, 70.9, 53.0, 51.3,
49.0, 48.2, 44.3, 43.1, 37.6, 35.7, 35.3, 34.6, 31.7, 29.5,
27.2, 26.2, 24.9, 22.3, 19.5, 17.3, 17.0, 13.9. ESI-HRMS
3
.70 (m, 6H), 1.65 (m, 1H), 1.42 (m, 1H), 1.23 (brt, J =
2.7 Hz, 2H), 1.08 (brt, J = 12.6 Hz, 1H), 1.03 (d, J = 6.2
+
Hz, 3H, CH ), 1.00-0.94 (m, 2H), 0.86 (d, J = 6.9 Hz, 3H,
(m/z): calcd. for C H N O [M+H] : 641.3914; found,
3
35 53
4
7
CH ), 0.84 (d, J = 6.2 Hz, 3H, CH ), 0.83 (d, J = 6.2 Hz, 3H,
641.3904.
3
3
13
CH ), 0.71 (brt, J = 12.6 Hz, 1H); C NMR (150 MHz,
3
4
.1.2.18. (1R,2R)-N-[2-(1H-indol-2-yl)ethyl]-2-
CD OD) δ (ppm): 177.6, 172.4, 171.7, 144.2, 138.9, 127.4,
3
[(2S,4E,6Z,8R,9S, 11R,13S,15S,16S)-7-cyano-8,16-
1
4
2
18.5, 116.0, 76.2, 71.6, 71.3, 51.6, 51.3, 48.8, 47.9, 44.2,
3.1, 37.6, 37.0, 35.7, 35.3, 34.5, 31.6, 30.5, 29.9, 29.8,
dihydroxy-9,11,13,15-tetrameth-yl-18-
oxooxacyclooctadeca-4,6-dien-2-yl]cyclopentane-1-
carboxamide (2r)
7.0, 26.2, 25.1, 19.5, 17.7, 17.2, 14.0, 14.0. ESI-HRMS
+
(m/z): calcd. for C H N O S [M+H] : 635.3730; found,
34
55
2
7
6
35.3732.
2
0
4
.1.2.16. methyl{(1R,2R)-2-
[(2S,4E,6Z,8R,9S,11R,13S,15S,16S)-7-cyano-8,16-
2
1
dihydroxy-9,11,13,15-tetramethyl-18-oxooxacyclo-
octadeca-4,6-dien-2-yl]cyclopentane-1-carbonyl}-D-
leucinate (2p)
3
Eluent petroleum ether/EtOAc (2:1). Colorless crystal.
5% yield: []
2
0
8
D
-36.4 (c 0.33, MeOH); UV (MeOH) λmax
(log ε) 256 (4.63) nm; IR v 3371, 2957, 2925, 2875, 2854,
2
1
213, 1727, 1655, 1534, 1463, 1378, 1281, 1260, 1204,
176, 1095, 1041, 1023, 972, 802, 745, 651 cm ; H NMR
(dt, J =10.2, 3.0 Hz, 1H), 3.84 (s, 3H, -OCH ), 3.61 (dt, J
3
-1
1
= 13.2, 7.5 Hz, 1H), 3.32 (dt, J = 13.2, 7.5 Hz, 1H), 2.91 (t,
J = 7.5 Hz, 2H), 2.73 (m, 1H), 2.58 (dd, J = 14.9, 3.9 Hz,
1H), 2.52 (brd, J = 14.9 Hz, 1H), 2.37 (m, 1H), 2.34 (dd, J
(600 MHz, CD OD) δ (ppm): 6.90 (d, J = 11.3 Hz, 1H),
3
6
.60 (dd, J = 14.9, 11.3 Hz, 1H), 6.32 (ddd, J = 14.9, 10.5,

=
16.3, 2.9 Hz, 1H), 2.25 (dd, J = 16.3, 10.2 Hz, 1H), 2.01
oxooxacyclooctadeca-4,6-dien-2-yl]cyclopentane-1-
carboxylate (3a)
(m, 1H), 1.94 (m, 1H), 1.88-1.76 (m, 4H), 1.72 (m, 2H)
1
1
0
0
0
.63 (m, 1H), 1.42 (m, 1H), 1.21(brt, J = 12.6 Hz, 1H),
.13(brt, J = 12.1 Hz, 1H), 1.04 (m, 1H), 0.91 (m, 2H),
.67 (brt, J = 11.6 Hz, 1H), 1.04 (d, J = 6.3 Hz, 3H, CH ),
Eluent dichloromethane/methanol (50:1). White crystal.
2
0
7
(
5% yield: []
log ε) 254 (4.52) nm; IR v 3419, 2956, 2922, 2211, 1730,
1638, 1595, 1512, 1459, 1375, 1282, 1168, 1081, 1040,
D
-12.6 (c 0.95, MeOH); UV (MeOH) λmax
3
.83 (d, J = 6.3 Hz, 3H, CH ), 0.82 (d, J = 6.3 Hz, 3H, CH ),
3
3
13
.79 (d, J = 6.3 Hz, 3H, CH ); C NMR (150 MHz, CD OD)
-1
1
3
3
9
(
75, 871, 825, 750, 657 cm ; H NMR (600 MHz, CD OD) δ
3
δ (ppm): 177.2, 171.6, 153.6, 144.1, 139.1, 132.0, 127.6,
ppm): 7.69 (s, 1H, Ar-H), 7.17 (s, 1H, Ar-H), 7.00 (s, 1H,
Ar-H), 6.94 (d, J = 11.3 Hz, 1H), 6.59 (dd, J = 15.0, 11.3
Hz, 1H), 6.29 (ddd, J = 14.9, 10.4, 4.7 Hz, 1H), 4.96 (dt, J
1
7
3
1
27.4, 122.7, 118.5, 116.0, 111.5, 111.4, 111.2, 99.9, 76.4,
1.6, 71.3, 54.9, 50.3, 48.2, 44.6, 43.2, 40.1, 37.6, 37.0,
5.6, 35.4, 34.5, 31.8, 29.3, 27.0, 26.2, 25.2, 24.7, 19.4,
7.7, 17.2, 13.9. ESI-HRMS (m/z): calcd. For C H N O
=
10.2, 3.6 Hz, 1H), 4.44 (m, 1H, -CO-O-CH -), 4.36-4.26
a
39
56
3
6
+
(m, 3H), 4.19 (d, J = 9.6 Hz, 1H), 3.87 (dt, J = 9.8, 3.3 Hz,
H), 2.63 (m, 1H), 2.59-2.51 (m, 3H), 2.29 (dd, J = 15.8,
[M+H] : 662.4169; found, 662.4167.
1
4
.1.2.19. (1R,2R)-2-[(2S,4E,6Z,8R,9S,11R,13S,15S,16S)-7-
3.0 Hz, 1H), 2.19 (dd, J = 15.8, 10.2 Hz, 1H), 2.01-1.92 (m,
2H), 1.84-1.77 (m, 3H), 1.77-1.72 (m, 3H), 1.64 (m, 1H),
1.42 (m, 1H), 1.22 (brt, J = 13.0 Hz, 1H), 1.17 (brt, J =
11.7 Hz, 1H), 1.08 (brt, J = 12.4 Hz, 1H), 0.97 (m, 1H),
cyano-8,16-dihydroxy-9,11,13,15-tetramethyl-18-
oxooxacyclooctadeca-4,6-dien-2-yl]-N-(4-
methoxyphenethyl)cyclopentane-1-carbox-amide (2s)
0
.94 (m, 1H), 0.70 (brt, J = 12.4 Hz, 1H), 1.04 (d, J = 6.4
Hz, 3H, CH ), 0.85 (d, J = 6.4 Hz, 3H, CH ), 0.84 (d, J =
2
0
3
3
13
6
1
.9 Hz, 6H, 2 × CH ); C NMR (150 MHz, CD OD) δ (ppm):
3 3
75.7, 171.5, 143.9, 138.5, 127.8, 127.5, 119.4, 118.4,
2
1
116.0, 75.8, 71.5, 71.1, 63.3, 48.4, 47.9, 45.5, 45.4 , 43.1,
3
3
1
7.6, 37.1, 35.7, 35.0, 34.5, 30.7, 28.9, 27.1, 26.2, 24.7,
9.4, 17.3, 17.3, 13.9. ESI-HRMS (m/z): calcd. For
+
C H N O [M+H] : 584.3700; found, 584.3700.
33
51
3
6
=
10.7, 3.6 Hz, 1H), 4.19 (d, J = 9.8 Hz, 1H), 3.91 (dt, J =
4.1.3.2. 2-(pyrrolidin-1-yl)ethyl (1R,2R)-2-
1
7
2
1
0.5, 3.1 Hz, 1H), 3.78 (s, 3H, -OCH ), 3.51 (dt, J = 13.2,
[(2S,4E,6Z,8R,9S,11R, 13S,15S,16S)-7-cyano-8,16-
dihydroxy-9,11,13,15-tetramethyl-18-oxooxacyclooctadeca-
4,6-dien-2-yl]cyclopentane-1-carboxyl-ate (3b)
3
.5 Hz, 1H), 3.18 (dt, J = 13.2, 7.5 Hz, 1H), 2.72 (m, 3H),
.60 (dd, J = 14.9, 3.9 Hz, 1H), 2.54 (brd, J = 14.9 Hz,
H), 2.36 (dd, J = 16.1, 3.0 Hz, 1H), 2.34 (m, 1H), 2.26
Eluent petroleum ether/EtOAc (2:1, 1:3). Colorless
(dd, J = 16.1, 10.2 Hz, 1H), 2.01 (m, 1H), 1.94 (m, 1H),
2
0
crystal. 69% yield: []
D
-18.5 (c 0.86, MeOH); UV (MeOH)
1
1
1
1
.82 (m, 4H), 1.74 (m, 1H), 1.70-1.60 (m, 2H), 1.43 (m,
H), 1.22 (brt, J = 11.7 Hz, 1H), 1.15 (brt, J = 12.1 Hz,
H), 1.06 (brt, J = 12.6 Hz, 1H), 0.96 (m, 1H), 0.91 (m,
H), 0.69 (brt, J = 12.6 Hz, 1H), 1.04 (d, J = 6.3 Hz, 3H,
λmax (log ε) 256 (4.52) nm; IR v 3644, 3440, 2958, 2923,
2
9
6
878, 2214, 1726, 1637, 1460, 1378, 1281, 1170, 1035,
75, 847, 739 cm ; H NMR (600 MHz, CD OD) δ (ppm):
.91 (d, J = 11.3 Hz, 1H), 6.61 (dd, J = 14.9, 11.3 Hz, 1H),
-1
1
3
CH ), 0.85 (d, J = 6.3 Hz, 3H, CH ), 0.84 (d, J = 7.0 Hz, 3H,
3
3
1
3
6.34 (ddd, J = 14.9, 10.4, 4.7 Hz, 1H), 4.97 (dt, J = 10.5,
.7 Hz, 1H), 4.34 (ddd, J = 11.7, 6.4, 5.3 Hz, 1H), 4.19 (d,
J = 9.8 Hz, 1H), 4.12 (ddd, J = 11.7, 6.4, 5.3 Hz, 1H), 3.91
dt, J = 10.1, 3.2 Hz, 1H), 2.83-2.62 (m, 6H, 3 × -CH -N-),
CH ), 0.83 (d, J = 6.3 Hz, 3H, CH ); C NMR (150 MHz,
CD OD) δ (ppm): 177.1, 171.6, 158.4, 144.1, 139.0, 130.8,
3
3
3
3
1
7
3
1
29.3 (2C), 127.4, 118.5, 116.0, 113.5 (2C), 76.4, 71.6,
1.3, 54.3, 50.3, 48.2, 44.5, 43.2, 41.1, 37.6, 37.1, 35.6,
5.4, 34.5, 34.5, 31.8, 29.3, 27.0, 26.2, 24.7, 19.4, 17.7,
7.2, 13.9. ESI-HRMS (m/z): calcd. For C H N O Na
(
2
2
.64 (m, 1H), 2.60-2.50 (m, 3H), 2.33 (dd, J = 15.9, 3.0
Hz, 1H), 2.21 (dd, J = 15.9, 10.0 Hz, 1H), 2.05-1.97 (m,
2H), 1.91-1.75 (m, 10H, 2 × -CH -pyrrole), 1.66 (m, 1H),
.43 (m, 1H), 1.25 (brt, J = 12.0 Hz, 1H), 1.17(brt, J =
3
7
55
2
6
+
2
[M+H] : 623.4060; found, 623.4062.
1
4
3
.1.3. General Procedure for the Synthesis of Compounds
a-3i
3
3
To a solution of borrelidin (40 mg, 0.082 mmol) in THF
2.0 mL) was added 1-hydroxybenzotriazole (HoBt, 22.2
mg, 0.164 mmoL) and N,N'-dicyclohexylcarbodiimide (DCC,
3.8 mg, 0.164 mmol) at room temperature. The reaction
13
(
3
mixture was stirred at room temperature for 0.5 h after
which the appropriate alcohol (0.205 mmol) was added.
Keep stirring for 12-24 h, the mixture was extracted with
EtOAc (20 mL). The organic layer was then washed with 3% 4.1.3.3. 2-(4-methylpiperazin-1-yl)ethyl(1R,2R)-2-
HCl aqueous (2 × 10 mL), saturated NaHCO aqueous (2 ×
[(2S,4E,6Z,8R, 9S,11R,13S,15S,16S)-7-cyano-8,16-
dihydroxy-9,11,13,15-tetra-methyl-18-
oxooxacyclooctadeca-4,6-dien-2-yl]cyclopentane-1-
carboxylate (3c)
3
1
0 mL), brine (2 × 10 mL). The resulting organic layer
wasdried over Na SO , filtered and concentrated in vacuo.
2
4
The residue was purified by silica gel column
chromatography eluting with appropriate mixture as
indicated in each case. The data of compound 3e and 3f
can be found at the literature.
Eluent dichloromethane/methanol (25:1). Yellow crystal.
2
0
41,42
65% yield: []
log ε) 254 (4.72) nm; IR v 3642, 3477, 3410, 2924, 2211,
1724, 1647, 1543, 1447, 1372, 1259, 1180, 1104, 1038,
D
-13.3 (c 0.30, MeOH); UV (MeOH) λmax
(
4
.1.3.1. 2-(1H-imidazol-1-yl)ethyl(1R,2R)-2-
-1
1
[(2S,4E,6Z,8R,9S, 11R,13S,15S,16S)-7-cyano-8,16-
975, 850, 769, 699, 638 cm ; H NMR (600 MHz, CD OD) δ
3
dihydroxy-9,11,13,15-tetra-methyl-18-
(ppm): 6.92 (d, J = 11.3 Hz, 1H), 6.60 (dd, J = 14.9, 11.4

Hz, 1H), 6.32 (ddd, J = 14.9, 10.4, 4.7 Hz, 1H), 4.97 (dt, J
10.5, 3.7 Hz, 1H), 4.29 (dt, J = 11.6, 5.7 Hz, 1H), 4.18
26.2, 24.8, 19.5, 17.7, 17.3, 14.0. ESI-MS (m/z): 561.4000
+
=
[M+H] , calcd. For C H N O .
3
2
53
2
6
d, J = 9.8 Hz, 1H), 4.14 (dt, J = 11.6, 5.7 Hz, 1H), 3.89
4
[
.1.3.6. 2-morpholinoethyl (1R,2R)-2-
(2S,4E,6Z,8R,9S,11R,13S, 15S,16S)-7-cyano-8,16-
dihydroxy-9,11,13,15-tetramethyl-18-oxooxacyclooctadeca-
2
.60-2.50 (m, 4H), 2.32 (dd, J = 15.9, 3.0 Hz, 1H), 2.21
4
,6-dien-2-yl]cyclopentane-1-carboxylate (3f)
1
1
H), 1.17 (brt, J = 12.3 Hz, 1H), 1.10 (brt, J = 11.5 Hz,
H), 0.97 (m, 2H), 0.69 (brt, J = 11.5 Hz, 1H), 2.49 (s, 3H,
Eluent dichloromethane/methanol (80:1). Colorless
2
0
crystal. 83% yield: [] -22.4 (c 1.07, MeOH); UV (MeOH)
D
-
NCH ), 1.03 (d, J = 6.4 Hz, 3H, CH ), 0.86 (d, J = 6.7 Hz,
λmax (log ε) 252 (4.49) nm; IR v 3435, 2954, 2923, 2872,
3
3
1
3
6
H, 2 × CH ), 0.85 (d, J = 6.4 Hz, 3H, CH ); C NMR (150
2212, 1728, 1639, 1457, 1375, 1270, 1169, 1118, 1035,
3
3
-1
1
MHz, CD OD) δ (ppm): 176.2, 171.6, 143.9, 138.7, 127.6,
976, 862, 772, 689 cm ; H NMR (600 MHz, CD OD) δ
3
3
1
(
3
18.5, 116.2, 76.0, 71.5, 71.4, 61.4, 56.0, 54.1 (2C), 51.7
2C), 48.7, 48.1, 45.6, 43.9, 43.3, 37.7, 37.0, 35.7, 35.2,
4.6, 30.8, 29.1, 27.3, 26.3, 24.8, 19.5, 17.6, 17.3, 14.0.
(ppm): 6.91 (d, J = 11.3 Hz, 1H), 6.61 (dd, J = 14.9, 11.3
Hz, 1H), 6.32 (ddd, J = 14.9, 10.4, 4.7 Hz, 1H), 4.97 (dt, J
= 10.3, 3.6 Hz, 1H), 4.35 (ddd, J = 11.7, 6.4, 5.1 Hz, 1H),
4.18 (d, J = 9.8 Hz, 1H), 4.11 (ddd, J = 11.7, 6.4, 5.1 Hz,
1H), 3.91 (dt, J = 10.3, 3.1 Hz, 1H), 3.69 (t, J = 4.7 Hz,
+
ESI-HRMS (m/z): calcd. For C H N O [M+H] : 616. 4326;
found, 616.4327.
35
58
3
6
H, 2 × -CH -O-), 2.71 (m, 1H), 2.63 (m, 2H), 2.57 (m,
2
.1.3.4. 2-(piperidin-1-yl)ethyl(1R,2R)-2-
H), 2.52 (m, 6H, 3 × -CH -N-), 2.32 (dd, J = 16.0, 3.0 Hz,
2
H), 2.21 (dd, J =16.0, 10.2 Hz, 1H), 2.00 (m, 2H), 1.85
,6-dien-2-yl]cyclopentane-1-carboxyl-ate (3d)
(brt, J = 12.5 Hz, 1H), 1.00 (m, 1H), 0.96 (m, 1H), 0.70
2
0
(brt, J = 12.5 Hz, 1H), 1.03 (d, J = 6.4 Hz, 3H, CH ), 0.85
3
(d, J = 6.4 Hz, 6H, 2 × CH ), 0.84 (d, J = 6.4 Hz, 3H, CH );
3
3
13
2
9
(
C NMR (150 MHz, CD OD) δ (ppm): 176.2, 171.6, 144.0,
3
138.7, 127.6, 118.5, 116.2, 76.0, 71.5, 71.5, 66.3 (2C),
61.3, 56.9, 53.6 (2C), 53.4, 48.7, 45.6, 43.3, 37.7, 36.8,
35.7, 35.2, 34.6, 30.8, 29.1, 27.2, 26.3, 24.9, 19.5, 17.8,
+
=
17.3, 14.0. ESI-HRMS (m/z): calcd. For C H N O [M+H] :
34 55 2 7
H, -CO-O-CH -), 4.21 (d, J = 9.8 Hz, 1H), 3.92 (dt, J =
b
0.1, 3.2 Hz, 1H), 3.21-3.01 (m, 6H, 3 × -CH -N-), 2.75 (m,
2
4
H), 2.62 (m, 1H), 2.60 (m, 1H), 2.58 (m, 1H), 2.36 (brd, J
15.9 Hz, 1H), 2.25 (dd, J =15.9, 10.0 Hz, 1H), 2.10-1.98
2
H, 1 × -CH -piperidine), 1.47 (m, 1H), 1.27 (m, 1H), 1.21
2
(
m, 1H), 1.13 (m, 1H), 0.98 (m, 1H), 0.92 (m, 1H), 0.70
Eluent petroleum ether/EtOAc (1:3). Colorless crystal.
2
0
(
brt, J = 11.6 Hz, 1H), 1.06 (d, J = 6.4 Hz, 3H, CH ), 0.89
88% yield: [] -20.4 (c 0.98, MeOH); UV (MeOH) λmax
3
D
(d, J = 6.4 Hz, 3H, CH ), 0.87 (d, J = 6.4 Hz, 3H, CH ),
(log ε) 202 (4.15), 256 (4.85) nm; IR v 3469, 3421, 2954,
3
3
13
0
.86 (d, J = 6.4 Hz, 3H, CH ); C NMR (150 MHz, CD OD)
2923, 2213, 1729, 1640, 1593, 1453, 1378, 1270, 1167,
3
3
-1
1
δ (ppm): 175.6, 171.6, 143.9, 138.8, 127.5, 118.5, 116.2,
1096, 1037, 974, 862, 817, 764, 670 cm ; H NMR (600
7
3
2
6.0, 71.6, 71.3, 59.4, 55.9, 53.8 (2C), 48.4, 45.4, 43.2,
7.6, 37.2, 35.7, 35.3, 34.5, 30.7, 29.1, 27.2, 26.2, 24.8,
MHz, CD OD) δ (ppm): 8.43 (d, J = 5.0 Hz, 1H, Ar-H), 7.75
3
(td, J = 7.7, 1.8 Hz, 1H, Ar-H), 7.39 (d, J = 7.7 Hz, 1H, Ar-
H), 7.27 (dd, J = 7.7, 5.0 Hz, 1H, Ar-H), 6.82 (d, J = 11.3
Hz, 1H), 6.51 (dd, J = 14.9, 11.3 Hz, 1H), 6.25 (ddd, J =
3.7 (2C), 22.0, 19.5, 17.5, 17.2, 13.9. ESI-HRMS (m/z):
+
calcd. For C H N O [M+H] : 601. 4217; found, 601.4216.
35
57
2
6
1
5
4.9, 10.4, 4.7 Hz, 1H), 5.18 (d, J = 13.7 Hz, 1H, -CH -Py),
a
.1.3.5. 2-(dimethylamino)ethyl(1R,2R)-2-[(2S,4E,6Z,8R,9S,
1R,13S,15S,16S)-7-cyano-8,16-dihydroxy-9,11,13,15-
.03 (d, J = 13.7 Hz, 1H, -CH -Py), 4.90 (dt, J = 10.8, 4.2
b
Hz, 1H), 4.08 (d, J = 9.8 Hz, 1H), 3.75 (dt, J = 10.4, 3.1
Hz, 1H), 2.69 (m, 1H), 2.53 (m, 1H), 2.48 (m, 1H), 2.44
(
m, 1H), 2.10 (dd, J = 15.9, 2.9 Hz, 1H), 2.00 (dd, J =
1
H NMR (600 MHz, CD OD) δ (ppm): 6.81 (d, J = 11.3
15.9, 10.3 Hz, 1H), 1.94 (m, 2H), 1.80 (m, 1H), 1.70 (m,
5H), 1.53 (m, 1H), 1.35 (m, 1H), 1.12 (brt, J = 13.5 Hz,
1H), 0.94 (m, 1H), 0.89 (m, 1H), 0.81 (m, 1H), 0.80 (m,
1H), 0.56 (brt, J = 12.5 Hz, 1H), 0.93 (d, J = 6.3 Hz, 3H,
CH ), 0.74(d, J = 6.7 Hz, 3H, CH ), 0.72 (d, J = 6.4 Hz, 3H,
3
4.9, 10.4, 4.7 Hz, 1H), 4.87 (dt, J = 10.5, 3.7 Hz, 1H),
.20 (ddd, J = 11.2, 6.2, 5.0 Hz, 1H), 4.08 (d, J = 9.8 Hz,
H), 4.02 (ddd, J = 11.2, 6.2, 5.0 Hz, 1H), 3.80 (dt, J =
0.1, 3.2 Hz, 1H), 2.64 (m, 1H), 2.58 (m, 2H), 2.50-2.41
3
3
1
3
CH ), 0.63 (d, J = 6.4 Hz, 3H, CH ); C NMR (150 MHz,
3
3
(
m, 3H), 2.25 (dd, J = 16.0, 2.9 Hz, 1H), 2.25 (s, 6H, 2 × -
NCH ), 2.11 (dd, J =16.0, 10.3 Hz, 1H), 1.92 (m, 2H),
.77-1.66 (m, 6H), 1.56 (m, 1H), 1.33 (m, 1H), 1.14 (brt, J
12.5 Hz, 1H), 1.07 (brt, J = 12.5 Hz, 1H), 1.00 (brt, J =
CD OD) δ (ppm): 175.8, 171.6, 155.7, 148.7, 144.1, 138.8,
3
137.6, 127.7, 123.2, 121.7, 118.5, 116.2, 76.0, 71.7, 71.5,
65.9, 48.5, 48.0, 45.8 , 43.2, 37.7, 36.6, 35.6, 35.2, 34.6,
30.9, 29.2, 27.2, 26.2, 25.0, 19.5, 17.9, 17.2, 14.0. ESI-
3
1
=
+
1
1
2
2.5 Hz, 1H), 0.84-0.91 (m, 2H), 0.59 (brt, J = 12.5 Hz,
HRMS (m/z): calcd. For C H N O [M+H] : 581.3591;
3
4
49
2
6
H), 0.93 (d, J = 6.4 Hz, 3H, CH ), 0.76 (d, J = 6.4 Hz, 6H,
found, 581.3592.
3
13
× CH ), 0.74 (d, J = 6.4 Hz, 3H, CH ); C NMR (150 MHz,
3
3
4
.1.3.8. phenethyl(1R,2S)-2-
CD OD) δ (ppm): 176.2, 171.6, 144.1, 138.9, 127.6, 118.5,
3
[(1R,3E,5Z,7R,8S,10R,12S,14S,15S)-6-cyano-7-hydroxy-
1
16.1, 76.0, 71.5, 71.5, 61.7, 57.2, 48.7, 48.1, 45.5, 44.5
(2C), 43.3, 37.7, 36.9, 35.7, 35.2, 34.6, 30.8, 29.1, 27.2,

8
3
,10,12,14,15-pentamethyl-17-methylene-cyclooctadeca-
,5-dien-1-yl]-2-methylcyclopentane-1-carboxyl-ate (3h)
washed with 3% HCl aqueous (2 × 50 mL), saturated
NaHCO aqueous (2 × 50 mL), brine (2 × 50 mL). The
3
resulting organic layer wasdried over Na SO , filtered and
2
4
Eluent petroleum ether/EtOAc (1:3). Colorless crystal.
concentrated in vacuo. The residue was purified by silica
gel column chromatography eluting with
dichloromethane/methanol (40:1) to form 1-Boc-4-(2-
hydroxyethyl)-piperazine a colorlessoil (1.592 g, 90% yield).
2
0
8
0% yield: []
D
-22.9 (c 1.05, MeOH); UV (MeOH) λmax
(log ε) 206 (4.13), 256 (4.50) nm; IR v 3486, 3447, 2957,
2
1
922, 2213, 1725, 1638, 1597, 1458, 1372, 1262, 1173,
-1 1
036, 975, 746, 698, 643 cm ; H NMR (600 MHz, CD OD)
3
δ (ppm): 7.20 (m, 2H, Ar-H), 7.14 (m, 3H, Ar-H), 6.79 (d, J
11.3 Hz, 1H), 6.50 (dd, J = 14.9, 11.3 Hz, 1H), 6.16
ddd, J = 14.9, 10.4, 4.7 Hz, 1H), 4.85 (dt, J = 10.7, 3.6
Hz, 1H), 4.29 (dt, J = 10.8, 6.9 Hz, 1H, -CO-O-CH -), 4.09
To a solution of BN (500 mg, 1.022 mmol) in THF (12
mL) was added 1-hydroxybenzotriazole (HoBt, 276.206 mg,
2.044 mmoL) and N,N'-dicyclohexylcarbodiimide (DCC,
421.739 mg, 2.044 mmol) at room temperature. After
stirring at room temperature for 0.5 h, the 1-Boc-4-(2-
hydroxyethyl)-piperazine (588.059 mg, 2.555 mmol) was
added. After another for 12 h, the excess THF was
evaporated, and then the mixture was extracted with
EtOAc (2 × 50 mL), and washed with 3% HCl aqueous (2
=
(
a
(
dt, J = 10.8, 6.9 Hz, 1H, -CO-O-CH -), 4.08 (d, J = 9.8 Hz,
b
1
2
2
H), 3.80 (dt, J = 10.3, 3.1 Hz, 1H), 2.83 (brt, J = 6.8 Hz,
H), 2.54 (m, 1H), 2.46 (ddd, J = 14.7, 10.5, 4.0 Hz, 1H),
.40 (brd, J = 14.7 Hz, 1H), 2.37 (q, J = 7.5 Hz, 1H), 2.21
(dd, J = 16.0, 2.9 Hz, 1H), 2.09 (dd, J = 16.0, 10.3 Hz,
1
1
=
H), 1.86 (m, 2H), 1.72 (m, 3H), 1.65 (m, 3H), 1.52 (m,
H), 1.30 (m, 1H), 1.11 (brt, J = 12.5 Hz, 1H), 1.03 (brt, J
12.8 Hz, 1H), 0.90 (m, 1H), 0.86 (brt, J = 12.5 Hz, 1H),
× 50 mL), saturated NaHCO aqueous (2 × 50 mL), brine
3
(2 × 50 mL). The resulting organic layer was dried over
Na SO , filtered and concentrated in vacuo. The residue
2
4
0
0
0
.78 (brt, J = 12.0 Hz, 1H), 0.57 (brt, J = 12.3 Hz, 1H),
.92 (d, J = 6.4 Hz, 3H, CH ), 0.74 (d, J = 6.0 Hz, 3H, CH ),
was purified by silica gel column chromatography eluting
with petroleum ether/EtOAc (2:1) to afford target
compound 4a as a colorless crystal (551.957 mg, 77%
yield).
3
3
.73 (d, J = 6.0 Hz, 3H, CH ), 0.69 (d, J = 6.5 Hz, 3H, CH );
3
3
1
3
C NMR (150 MHz, CD OD) δ (ppm): 176.3, 171.6, 144.1,
3
1
1
3
1
38.7, 137.9, 128.6 (2C), 128.2 (2C), 127.6, 126.2, 118.5,
16.1, 76.0, 71.6, 71.5, 64.9, 48.9, 47.9, 45.6, 43.2, 37.6,
6.7, 35.7, 35.1, 34.8, 34.6, 30.7, 29.1, 27.1, 26.2, 24.8,
The solution of 4a (500 mg, 0.713 mmol) in CH Cl (5
2
2
mL) was cooled to 0℃ and TFA (5mL) was slowly added
and stirring was continued for 30 min (monitored by TLC),
then quenched by the addition of saturated aqueous
Na CO . The mixture was extracted with CH Cl (2 × 50
9.4, 17.8, 17.3, 14.0. ESI-HRMS (m/z): calcd. For
+
C H NO Na [M+Na] : 616.3614; found, 616.3614.
36
50
5
2
3
2
2
4
[
.1.3.9. cyclopropylmethyl(1R,2R)-2-
(2S,4E,6Z,8R,9S,11R,13S, 15S,16S)-7-cyano-8,16-
dihydroxy-9,11,13,15-tetramethyl-18-oxooxacyclooctadeca-
mL), washed with brine, dried over anhydrous Na SO ,
2 4
filtered and concentrated in vacuo. The residue was further
purified by silica gel column chromatography eluting with
dichloromethane /methanol (25: 1) to prepare compound
4
,6-dien-2-yl]cyclopentane-1-carboxylate (3i)
4
b as a colorless crystal (372.928 mg, 87% yield).
Eluent petroleum ether/EtOAc (1:1). Colorless crystal.
2
0
7
(
1
5% yield: []
D
-12.6 (c 0.95, MeOH); UV (MeOH) λmax
To a solution of 4b (20 mg, 0.033 mmol) in CH CN (1.5
3
log ε) 256 (4.63) nm; IR v 3455, 2925, 2213, 1724, 1639,
mL) was added potassium carbonate (4.5621 mg, 0.033
mmol), appropriate halohydrocarbon (0.028 mmol) and
sodium iodide (catalytic amount). The mixture was heated
-
1
1
457, 1374, 1260, 1174, 1097, 1033, 976, 816 cm ; H
NMR (600 MHz, CD OD) δ (ppm): 6.81 (d, J = 11.3 Hz, 1H),
3
6
4
9
3
1
4
.51 (dd, J = 14.9, 11.3 Hz, 1H), 6.23 (ddd, J = 14.9, 10.4,
.7 Hz, 1H), 4.87 (dt, J = 10.7, 3.6 Hz, 1H), 4.09 (d, J =
at reflux for 1-2 h (monitored by TLC). Excess CH CN was
evaporated. Then the mixture was extracted with EtOAc (2
3
.8 Hz, 1H), 3.87 (dd, J = 11.4, 7.3 Hz, 1H, -CO-O-CH -),
× 10 mL), and washed with H O (2 × 10 mL) and brine (2
a
2
.81 (dt, J = 10.3, 3.0 Hz, 1H), 3.75 (dd, J = 11.4, 7.3 Hz,
× 10 mL), dried with anhydrous Na SO , filtered and
2
4
H, -CO-O-CH -), 2.62 (m, 1H), 2.48 (ddd, J = 14.7, 10.5,
concentrated in vacuo. The residue was further purified by
silica gel column chromatography eluting with appropriate
mixture as indicated in each case to give target compounds
4c-4r.
b
.0 Hz, 1H), 2.43 (brd, J = 14.7 Hz, 1H), 2.38 (q, J = 8.0
Hz, 1H), 2.22 (dd, J = 16.0, 2.9 Hz, 1H), 2.09 (dd, J =16.0,
1
1
0
=
0
6
=
0.4 Hz, 1H), 1.91 (m, 2H), 1.76 (m, 2H), 1.70 (m, 4H),
.54 (m, 1H), 1.32 (m, 1H), 1.14 (brt, J = 13.0 Hz, 1H),
.96-1.06 (m, 3H), 0.90 (brt, J = 13.0 Hz, 1H), 0.84 (brt, J
4
.1.4.1. (2S,4E,6Z,8R,9S,11R,13S,15S,16S)-tert-butyl-4-{2-
[((1R, 2R)-2-(7-cyano-8,16-dihydroxy-9,11,13,15-
13.0 Hz, 1H), 0.59 (brt, J = 12.6 Hz, 1H), 0.46 (m, 2H),
tetramethyl-18-oxo-oxacyclooctadeca-4,6-dien-2-
yl)cyclopentane-1-carbonyl)oxy]-ethyl}piperazine-1-
carboxylate (4a)
.19 (m, 2H), 0.93 (d, J = 6.4 Hz, 3H, CH ), 0.76 (d, J =
3
.4 Hz, 3H, CH ), 0.75 (d, J = 6.4 Hz, 3H, CH ), 0.74 (d, J
3
3
1
3
6.4 Hz, 3H, CH ); C NMR (150 MHz, CD OD) δ (ppm):
3
3
2
0
1
7
3
8
75.7, 170.7, 143.1, 137.8, 126.7, 117.5, 115.1, 75.0, 70.7,
0.6, 67.9, 47.9, 47.2, 44.7, 42.3, 36.7, 35.6, 34.7, 34.2,
3.6, 29.9, 28.1, 26.1, 25.3, 23.9, 18.6, 16.9, 16.3, 13.0,
.4, 1.4, 1.3. ESI-HRMS (m/z): calcd. For C H NO
[] -18.9 (c 1.06, MeOH); UV (MeOH) λmax (log ε)
D
256 (4.66) nm; IR v 3633, 3431, 2956, 2213, 1691, 1424,
-
1
1
1373, 1252, 1170, 1033, 978, 847 cm ; H NMR (600 MHz,
32
50
6
CD OD) δ (ppm): 6.81 (d, J = 11.3 Hz, 1H), 6.51 (dd, J =
3
+
[M+H] : 544.3638; found, 544.3638.
14.9, 11.3 Hz, 1H), 6.22 (ddd, J = 14.9, 10.3, 4.8 Hz, 1H),
4
1
1
.88 (dt, J = 10.6, 3.7 Hz, 1H), 4.22 (dt, J = 11.7, 5.9 Hz,
H), 4.09 (d, J = 9.8 Hz, 1H), 4.03 (dt, J = 11.7, 5.9 Hz,
H), 3.80 (dt, J = 10.1, 3.1 Hz, 1H), 3.33 (m, 4H, 2 × -
4
4
.1.4. General Procedure for the Synthesis of Compounds
a-4r
To a solution of 1-(2-hydroxyethyl) piperazine (1.0 g,
.686 mmol) in dichloromethane (25 mL) were added di-
CON-CH -), 2.61 (m, 2H), 2.55 (t, J = 5.0 Hz, 2H), 2.51-
2.41 (m, 3H, 1 × -CH -N-), 2.38 (m, 4H, 2 × -CH -N-), 2.23
2
7
2
2
tertbutyl dicarbonate (2.013 g, 9.223 mmol) and
triethylamine (TEA, 2.130 g, 15.372 mmol). The reaction
mixture was stirred at room temperature for 12 h, then
(dd, J = 15.9, 3.0 Hz, 1H), 2.12 (dd, J = 16.0, 10.1 Hz,
1H), 1.90 (m, 2H), 1.80-1.60 (m, 6H), 1.55 (m, 1H), 1.36
(s, 9H), 1.32 (m, 1H), 1.14 (brt, J = 12.6 Hz, 1H), 1.07 (brt,

J = 12.6 Hz, 1H), 1.00 (brt, J = 12.6 Hz, 1H), 0.88 (m, 2H),
30.9, 29.1, 27.2, 26.3, 24.9, 19.5, 17.7, 17.3, 14.0. ESI-
+
0
0
.60 (brt, J = 12.6 Hz, 1H), 0.94 (d, J = 6.4 Hz, 3H, CH ),
HRMS (m/z): calcd. For C H N O [M+H] : 736. 4537;
3
42 62
3
8
.76 (d, J = 6.4 Hz, 6H, 2 × CH ), 0.75 (d, J = 6.4 Hz, 3H,
found, 736.4536.
3
13
CH ); C NMR (150 MHz, CD OD) δ (ppm): 176.3, 171.6,
3
3
4
.1.4.4. 2-(4-benzoylpiperazin-1-yl) ethyl (1R,2R)-2-
1
7
4
54.9, 143.9, 138.7, 127.6, 118.5, 116.2, 79.9, 76.0, 71.6,
1.4, 61.3, 56.4, 52.9 (2C), 48.6 (2C), 48.2 (2C), 48.1,
5.6, 43.3, 37.7, 37.0, 35.7, 35.2, 34.6, 30.9, 29.1, 27.3
[(2S,4E,6Z, 8R,9S,11R,13S,15S,16S)-7-cyano-8,16-
dihydroxy-9,11,13,15-tetramethyl-18-oxooxacyclooctadeca-
4
,6-dien-2-yl] cyclopentane -1-carboxylate (4d)
(
3C), 27.2, 26.3, 24.8, 19.6, 17.7, 17.3, 14.0. ESI-HRMS
m/z): calcd. For C H N O [M+H] : 702.4693; found,
+
(
Eluent dichloromethane/methanol (50:1). Colorless
39
64
3
8
2
0
7
02.4693.
crystal. 95% yield: [] -44 (c 1.00, MeOH); UV (MeOH)
D
λmax (log ε) 252 (4.87) nm; IR v 3391, 2925, 2213, 1728,
4
.1.4.2. 2-(piperazin-1-yl) ethyl (1R,2R)-2-
1
626, 1448, 1375, 1273, 1168, 1025, 971, 857, 774, 712
[
(2S,4E,6Z,8R,9S,11R, 13S,15S,16S)-7-cyano-8,16-
dihydroxy-9,11,13,15-tetramethyl-18-oxooxacyclooctadeca-
,6-dien-2-yl]cyclopentane-1-carboxyl-ate (4b)
-1
1
cm ; H NMR (600 MHz, CD OD) δ (ppm): 7.37 (m, 3H, Ar-
3
H), 7.31 (m, 2H, Ar-H), 6.80 (d, J = 11.3 Hz, 1H), 6.51 (dd,
J = 14.9, 11.3 Hz, 1H), 6.22 (ddd, J = 14.9, 10.1, 4.8 Hz,
1H), 4.88 (dt, J = 10.5, 3.7 Hz, 1H), 4.22 (dt, J = 11.7, 5.7
Hz, 1H), 4.08 (d, J = 9.8 Hz, 1H), 4.05 (dt, J = 11.7, 5.7
Hz, 1H), 3.80 (dt, J = 10.1, 3.2 Hz, 1H), 3.67 (m, 2H, 1 × -
CON-CH -), 3.36 (m, 2H, 1 × -CON-CH -), 2.62 (m, 1H),
4
2
0
[]
D
-12 (c 1.00, MeOH); UV (MeOH) λmax (log ε) 256
(
1
4.49) nm; IR v 3419, 2926, 2219, 1681, 1451, 1379, 1199,
142, 1036, 973, 842, 801, 723, 669 cm ; H NMR (600
-1
1
MHz, CD OD) δ (ppm): 6.81 (d, J = 11.3 Hz, 1H), 6.51 (dd,
3
2
2
J = 14.9, 11.3 Hz, 1H), 6.22 (ddd, J = 14.9, 10.1, 4.8 Hz,
2.58 (t, J = 5.7 Hz, 2H, 1 × -N-CH -), 2.51-2.41 (m, 3H),
2
1
H), 4.88 (dt, J = 10.6, 3.7 Hz, 1H), 4.24 (m, 1H, -CO-O-
2.39 (m, 4H, 2 × -N-CH -), 2.22 (dd, J = 15.9, 3.1 Hz, 1H),
2
CH -), 4.11 (m, 1H, -CO-O-CH -), 4.09 (d, J = 9.7 Hz, 1H),
2.12 (dd, J = 15.9, 10.0 Hz, 1H), 1.91 (m, 2H), 1.72 (m,
3H), 1.67 (m, 3H), 1.54 (m, 1H), 1.33 (m, 1H), 1.14 (m,
1H), 1.08 (brt, J = 12.5 Hz, 1H), 0.99 (brt, J = 12.5 Hz,
1H), 0.89 (m, 1H), 0.85 (m, 1H), 0.59 (brt, J = 12.5 Hz,
a
b
3
2
.78 (dt, J = 9.9, 3.2 Hz, 1H), 3.19 (m, 4H, 2 × -NH-CH -),
2
.84 (m, 2H, 1 × -N-CH -), 2.78 (m, 4H, 2 × -N-CH -), 2.59
2
2
(
m, 1H), 2.53-2.43 (m, 3H), 2.23 (dd, J = 15.8, 3.1 Hz,
1
1
1
1
0
0
H), 2.14 (dd, J = 15.8, 9.8 Hz, 1H), 1.92 (m, 2H), 1.76-
.66 (m, 6H), 1.55 (m, 1H), 1.34 (m, 1H), 1.13 (brt, J =
2.6 Hz, 1H), 1.08 (brt, J = 12.6 Hz, 1H), 1.00 (brt, J =
2.6 Hz, 1H), 0.87 (m, 2H), 0.60 (brt, J = 12.6 Hz, 1H),
.94 (d, J = 6.4 Hz, 3H, CH ), 0.76 (d, J = 6.4 Hz, 3H, CH ),
1H), 0.93 (d, J = 6.4 Hz, 3H, CH ), 0.75 (d, J = 6.4 Hz, 3H,
3
CH ), 0.74 (d, J = 6.4 Hz, 3H, CH ), 0.73 (d, J = 6.4 Hz, 3H,
3
3
13
CH ); C NMR (150 MHz, CD OD) δ (ppm): 176.3, 171.6,
3
3
171.0, 143.9, 138.7, 135.3, 129.8, 128.4 (2C), 127.6,
126.7 (2C), 118.5, 116.2, 75.9, 71.6, 71.4, 61.3, 56.2, 53.3,
52.7, 48.6, 48.2, 45.6, 43.3, 41.8 (2C), 37.7, 37.0, 35.7,
35.2, 34.6, 30.9, 29.1, 27.2, 26.3, 24.8, 19.5, 17.6, 17.3,
3
3
.75 (d, J = 6.4 Hz, 3H, CH ), 0.74 (d, J = 6.4 Hz, 3H, CH );
3
3
1
3
C NMR (150 MHz, CD OD) δ (ppm): 176.2, 171.6, 144.0,
3
+
1
4
3
1
6
38.8, 127.5, 118.5, 116.1, 76.0, 71.6, 71.2, 60.6, 55.9,
9.6 (2C), 48.5, 48.4, 45.5, 43.2, 43.0 (2C), 37.7, 37.4,
5.8, 35.2, 34.6, 31.0, 29.1, 27.3, 26.3, 24.8, 19.5, 17.4,
14.0. ESI-HRMS (m/z): calcd. For C H N O [M+H] : 706.
41
60
3
7
4431; found, 706.4432.
+
4.1.4.5. 2-(4-acetylpiperazin-1-yl) ethyl (1R,2R)-2-
7.3, 14.0. ESI-HRMS (m/z): calcd. For C H N O [M+H] :
34
56
3
6
[
(2S,4E,6Z,8R, 9S,11R,13S,15S,16S)-7-cyano-8,16-
02.4169; found, 602.4168.
dihdihydroxy-9,11,13,15-tetramethyl-18-
oxooxacyclooctadeca-4,6-dien-2-yl]cyclopentane-1-
carboxylate (4e)
4
.1.4.3. benzyl-4-{2-[((1R,2R)-2
((2S,4E,6Z,8R,9S,11R,13S,15S, 16S)-7-cyano-8,16-
dihydroxy-9,11,13,15-tetramethyl-18-oxo-
oxacyclooctadeca-4,6-dien-2-yl)cyclopentane-1-
carbonyl)oxy]-ethyl}piperazine-1-carboxylate (4c)
2
0
1632, 1448, 1371, 1257, 1168, 1039, 987, 864, 764, 678
2
0
-1
1
cm ; H NMR (600 MHz, CD OD) δ (ppm): 6.91 (d, J =
3
11.3 Hz, 1H), 6.61 (dd, J = 14.9, 11.3 Hz, 1H), 6.32 (ddd,
J = 14.9, 10.1, 4.8 Hz, 1H), 4.97 (dt, J = 10.6, 3.7 Hz, 1H),
4.33 (dt, J = 11.5, 5.7 Hz, 1H), 4.18 (d, J = 9.8 Hz, 1H),
4.13 (dt, J = 11.5, 5.7 Hz, 1H), 3.90 (dt, J = 10.1, 3.1 Hz,
1
7
=
1H), 3.58 (t, J = 5.1 Hz, 2H, 1 × -CON-CH -), 3.54 (t, J =
2
1
4
4
1
=
4
H), 5.12 (s, 2H, -CH -Ar), 4.97 (dt, J = 10.6, 3.7 Hz, 1H),
.32 (dt, J = 11.7, 5.7 Hz, 1H), 4.18 (d, J = 9.8 Hz, 1H),
.11 (dt, J = 11.7, 5.7 Hz, 1H), 3.89 (dt, J = 10.2, 3.1 Hz,
5.1 Hz, 2H, 1 × -CON-CH -), 2.70 (m, 1H), 2.65 (t, J = 5.7
2
2
Hz, 2H), 2.59-2.49 (m, 7H), 2.33 (dd, J = 15.9, 3.0 Hz, 1H),
2.21 (dd, J = 15.9, 10.1 Hz, 1H), 2.09 (s, 3H), 2.00 (m,
2H), 1.87-1.77 (m, 6H), 1.65 (m, 1H), 1.42 (m, 1H), 1.23
(brt, J = 12.5 Hz, 1H), 1.17 (brt, J = 12.5 Hz, 1H), 1.10
(brt, J = 12.5 Hz, 1H), 1.00 (m, 1H), 0.96 (m, 1H), 0.69
H), 3.50 (m, 4H, 2 × -CON-CH -), 2.68 (m, 1H), 2.64 (t, J
2
5.9 Hz, 2H, 1 × -N-CH -), 2.60-2.52 (m, 3H), 2.50 (m,
2
H, 2 × -N-CH -), 2.32 (dd, J = 15.9, 3.0 Hz, 1H), 2.21 (dd,
2
J = 15.9, 10.1 Hz, 1H), 1.99 (m, 2H), 1.83 (m, 4H), 1.77
m, 2H), 1.64 (m, 1H), 1.42 (m, 1H), 1.23 (brt, J = 12.5 Hz,
(brt, J = 12.5 Hz, 1H), 1.03 (d, J = 6.4 Hz, 3H, CH ), 0.86
3
(
(d, J = 6.6 Hz, 3H, CH ), 0.85 (d, J = 5.4 Hz, 6H, 2 × CH );
3
3
13
1
1
6
=
H), 1.16 (brt, J = 12.5 Hz, 1H), 1.08 (brt, J = 12.5 Hz,
H), 0.97 (m, 2H), 0.69 (brt, J = 12.5 Hz, 1H), 1.03 (d, J =
.4 Hz, 3H, CH ), 0.85 (d, J = 6.4 Hz, 3H, CH ), 0.84 (d, J
C NMR (150 MHz, CD OD) δ (ppm): 176.2, 171.6, 170.2,
3
143.9, 138.7, 127.6, 118.5, 116.2, 75.9, 71.5, 71.4, 61.3,
56.3, 53.2, 52.7, 48.6, 48.1, 45.9, 45.6, 43.3, 41.1, 37.7,
37.0, 35.7, 35.2, 34.6, 30.9, 29.1, 27.2, 26.3, 24.8, 19.8,
19.5, 17.7, 17.3, 14.0. ESI-HRMS (m/z): calcd. For
3
3
13
6.4 Hz, 3H, CH ), 0.83 (d, J = 6.4 Hz, 3H, CH ); C NMR
3
3
(150 MHz, CD OD) δ (ppm): 176.2, 171.6, 155.4, 143.9,
3
+
1
1
4
38.7, 136.7, 128.1 (2C), 127.7, 127.6, 127.5 (2C), 118.5,
16.2, 76.0, 71.6, 71.4, 67.0, 61.3, 56.4, 52.8 (2C), 48.6,
8.1, 45.6, 43.5 (2C), 43.3, 37.7, 36.9, 35.7, 35.2, 34.6,
C H N O [M+H] : 644.4275; found, 644.4275.
36
58
3
7

4
[
.1.4.6. 2-(4-benzylpiperazin-1-yl) ethyl (1R,2R)-2-
(2S,4E,6Z, 8R,9S,11R,13S,15S,16S)-7-cyano-8,16-
dihydroxy-9,11,13,15-tetramethyl-18-oxooxacyclooctadeca-
oxooxacyclooctadeca-4,6-dien-2-yl)cyclopentane-1-
carboxylate (4h)
Eluent dichloromethane/methanol (50:1). Colorless
4
,6-dien-2-yl]cyclopentane-1-carboxylate (4f)
2
0
crystal. 90% yield: []
D
-21.8 (c 0.55, MeOH); UV (MeOH)
Eluent dichloromethane/methanol (50:1). Colorless
λmax (log ε) 256 (4.53) nm; IR v 3467, 2923, 2862, 2213,
2
0
crystal. 90% yield: []
D
-18.8 (c 0.64, MeOH); UV (MeOH)
1728, 1610, 1501, 1458, 1373, 1277, 1168, 1094, 1037,
-1
1
λmax (log ε) 254 (4.81) nm; IR v 3427, 2933, 2212, 1728,
636, 1457, 1373, 1284, 1166, 1033, 975, 822, 742, 694
971, 850, 812, 740 cm ; H NMR (600 MHz, CD OD) δ
3
1
(ppm): 7.44 (m, 1H, Ar-H), 6.94 (m, 2H, Ar-H), 6.92 (d, J
= 11.3 Hz, 1H), 6.60 (dd, J = 14.9, 11.3 Hz, 1H), 6.31
(ddd, J = 14.9, 10.1, 4.8 Hz, 1H), 4.96 (dt, J = 10.6, 3.7
Hz, 1H), 4.31 (dt, J = 11.5, 5.7 Hz, 1H), 4.18 (d, J = 9.8
Hz, 1H), 4.11 (dt, J = 11.5, 5.7 Hz, 1H), 3.89 (dt, J = 10.2,
-1
1
cm ; H NMR (600 MHz, CD OD) δ (ppm): 7.36-7.29 (m,
3
4
6
4
1
1
H, Ar-H), 7.26 (m, 1H, Ar-H), 6.93 (d, J = 11.3 Hz, 1H),
.60 (dd, J = 14.9, 11.3 Hz, 1H), 6.31 (ddd, J = 14.9, 10.1,
.8 Hz, 1H), 4.96 (dt, J = 10.6, 3.7 Hz, 1H), 4.30 (dt, J =
1.5, 5.9 Hz, 1H), 4.18 (d, J = 9.8 Hz, 1H), 4.12 (dt, J =
1.5, 5.9 Hz, 1H), 3.89 (dt, J = 10.2, 3.1 Hz, 1H), 3.57 (d,
3.1 Hz, 1H), 3.59 (s, 2H, -CH -Ar), 2.70 (m, 2H), 2.63 (m,
2
2H), 2.59 (m, 2H, 1 × -N-CH -), 2.57 (m, 2H, 1 × -N-CH -),
2
2
J = 12.8 Hz, 1H), 3.54 (d, J = 12.8 Hz, 1H), 2.70 (m, 2H),
.64 (m, 2H), 2.60 (m, 2H, 1 × -N-CH -), 2.56 (m, 2H, 1 ×
2.53 (m, 2H, 1 × -N-CH -), 2.49 (m, 4H, 2 × -N-CH -), 2.31
2
2
2
(dd, J = 15.9, 3.0 Hz, 1H), 2.20 (dd, J = 15.9, 10.2 Hz,
1H), 1.99 (m, 2H), 1.82 (m, 4H), 1.77 (m, 2H), 1.65 (m,
1H), 1.42 (m, 1H), 1.23 (brt, J = 12.5 Hz, 1H), 1.16 (brt, J
= 12.5 Hz, 1H), 1.09 (brt, J = 12.5 Hz, 1H), 0.99 (m, 1H),
0.96 (m, 1H), 0.69 (brt, J = 12.5 Hz, 1H), 1.03 (d, J = 6.4
Hz, 3H, CH ), 0.85 (d, J = 6.0 Hz, 3H, CH ), 0.84 (d, J =
2
-
N-CH -), 2.53 (m, 2H, 1 × -N-CH -), 2.49 (m, 4H, 2 × -N-
2
2
2
CH -), 2.31 (dd, J = 15.9, 3.0 Hz, 1H), 2.20 (dd, J = 15.9,
1
1
1
1
1
0
0.1 Hz, 1H), 1.98 (m, 2H), 1.82 (m, 4H), 1.77 (m, 2H),
.65 (m, 1H), 1.42 (m, 1H), 1.23 (brt, J = 13.0 Hz, 1H),
.16 (brt, J = 13.0 Hz, 1H), 1.09 (brt, J = 12.2 Hz, 1H),
.00 (m, 1H), 0.97 (m, 1H), 0.69 (brt, J = 12.2 Hz, 1H),
.03 (d, J = 6.5 Hz, 3H, CH ), 0.85 (d, J = 6.0 Hz, 3H, CH ),
3
3
13
6.0 Hz, 3H, CH ), 0.83 (d, J = 6.0 Hz, 3H, CH ); C NMR
3
3
(150 MHz, CD OD) δ (ppm): 176.2, 171.6, 162.3, 163.2,
3
3
3
13
.84 (d, J = 6.4 Hz, 6H, 2 × CH ); C NMR (150 MHz,
162.4, 162.3, 161.7, 161.6, 160.7, 160.6, 144.0, 138.7,
133.0, 127.6, 119.7, 118.5, 116.1, 110.7, 103.0, 76.0, 71.5
(2C), 61.4, 56.4, 54.0, 52.8 (2C), 52.9 (2C), 48.7, 48.1,
45.6, 43.3, 37.7, 36.9, 35.7, 35.2, 34.6, 30.8, 29.1, 27.2,
26.3, 24.8, 19.5, 17.7, 17.3, 14.0. ESI-HRMS (m/z): calcd.
3
CD OD) δ (ppm): 176.2, 171.6, 144.0, 138.7, 136.7, 129.4
3
(
2C), 127.9 (2C), 127.6, 127.1, 118.5, 116.2, 76.0, 71.5,
1.4, 62.5, 61.4, 56.4, 52.8 (2C), 52.2 (2C), 48.7, 48.1,
5.6, 43.3, 37.7, 36.9, 35.7, 35.2, 34.6, 30.8, 29.1, 27.2,
7
4
2
+
6.3, 24.8, 19.5, 17.7, 17.3, 14.0. ESI-HRMS (m/z): calcd.
For C H F N O [M+H] : 728.4450; found, 728.4450.
4
1
60
2
3
6
+
For C H N O [M+H] : 692.4639; found, 692.4641.
4
1
62
3
6
4.1.4.9. 2-[4-(4-chlorobenzyl)piperazin-1-yl] ethyl (1R,2R)-
4
.1.4.7. 2-[4-(4-fluorobenzyl) piperazin-1-yl] ethyl (1R,2R)-
-[(2S,4E,6Z,8R,9S,11R,13S,15S,16S)-7-cyano-8,16-
2-[(2S,4E,6Z,8R,9S,11R,13S,15S,16S)-7-cyano-8,16-
dihydroxy-9,11,13,15-tetramethyl-18-oxooxacyclooctadeca-
4,6-dien-2-yl]-cyclopentane-1-carboxylate (4i)
2
dihydroxy-9,11,13,15-tetramethyl-18-oxooxacyclooctadeca-
4
,6-dien-2-yl]cyclopentane-1-carboxylate (4g)
2
0
Eluent dichloromethane/methanol (50:1). Colorless
2
0
crystal. 90% yield: []
D
-21.8 (c 0.55, MeOH); UV (MeOH)
λmax (log ε) 258 (4.40) nm; IR v 3393, 2930, 2212, 1727,
1636, 1595, 1457, 1374, 1291, 1167, 1092, 1031, 974, 806,
-1
1
1
7
=
636, 1510, 1456, 1372, 1282, 1223, 1165, 1035, 974, 841,
671 cm ; H NMR (600 MHz, CD OD) δ (ppm): 7.32 (m, 4H,
3
-1
1
64 cm ; H NMR (600 MHz, CD OD) δ (ppm): 7.35 (dd, J
Ar-H), 6.93 (d, J = 11.3 Hz, 1H), 6.60 (dd, J = 14.9, 11.3
Hz, 1H), 6.31 (ddd, J = 14.9, 10.1, 4.8 Hz, 1H), 4.96 (dt, J
= 10.7, 3.7 Hz, 1H), 4.31 (dt, J = 11.5, 5.5 Hz, 1H), 4.18
(d, J = 9.8 Hz, 1H), 4.11 (dt, J = 11.5, 5.5 Hz, 1H), 3.89
(dt, J = 10.2, 3.1 Hz, 1H), 3.54 (d, J = 13.0 Hz, 1H), 3.51
(d, J = 13.0 Hz, 1H), 2.70 (m, 2H), 2.63 (m, 2H), 2.60 (m,
2H, 1 × -N-CH -), 2.58 (m, 2H, 1 × -N-CH -), 2.53 (m, 2H,
3
8.5, 5.6 Hz, 2H, Ar-H), 7.04 (brt, J = 8.5 Hz, 2H, Ar-H),
6
6
3
9
1
1
.93 (d, J = 11.3 Hz, 1H), 6.60 (dd, J = 14.9, 11.3 Hz, 1H),
.31 (ddd, J = 14.9, 10.1, 4.8 Hz, 1H), 4.96 (dt, J = 10.6,
.7 Hz, 1H), 4.31 (dt, J = 11.5, 5.7 Hz, 1H), 4.18 (d, J =
.8 Hz, 1H), 4.12 (dt, J = 11.5, 5.7 Hz, 1H), 3.89 (dt, J =
0.2, 3.1 Hz, 1H), 3.56 (d, J = 13.0 Hz, 1H), 3.53 (d, J =
3.0 Hz, 1H), 2.70 (m, 2H), 2.64 (m, 2H), 2.60 (m, 2H, 1 ×
2
2
1 × -N-CH -), 2.49 (m, 4H, 2 × -N-CH -), 2.31 (dd, J =
2
2
-
N-CH -), 2.57 (m, 2H, 1 × -N-CH -), 2.53 (m, 2H, 1 × -N-
15.9, 3.0 Hz, 1H), 2.20 (dd, J = 15.9, 10.2 Hz, 1H), 1.98
(m, 2H), 1.83 (m, 3H), 1.77 (m, 3H), 1.65 (m, 1H), 1.41
(m, 1H), 1.23 (brt, J = 12.6 Hz, 1H), 1.16 (brt, J = 12.6 Hz,
1H), 1.08 (brt, J = 12.6 Hz, 1H), 0.98 (m, 1H), 0.96 (m,
1H), 0.69 (brt, J = 12.6 Hz, 1H), 1.03 (d, J = 6.4 Hz, 3H,
CH ), 0.85 (d, J = 6.0 Hz, 3H, CH ), 0.84 (d, J = 6.0 Hz, 3H,
2
2
CH -), 2.49 (m, 4H, 2 × -N-CH -), 2.31 (dd, J = 15.9, 3.0
Hz, 1H), 2.20 (dd, J = 15.9, 10.1 Hz, 1H), 1.98 (m, 2H),
2
2
1
1
1
0
0
0
.82 (m, 4H), 1.77 (m, 2H), 1.65 (m, 1H), 1.41 (m, 1H),
.23 (brt, J = 12.5 Hz, 1H), 1.16 (brt, J = 12.5 Hz, 1H),
.09 (brt, J = 12.5 Hz, 1H), 0.99 (m, 1H), 0.97 (m, 1H),
3
3
1
3
.69 (brt, J = 12.5 Hz, 1H), 1.03 (d, J = 6.4 Hz, 3H, CH ),
CH ), 0.82 (d, J = 6.0 Hz, 3H, CH ); C NMR (150 MHz,
3
3
3
.85 (d, J = 6.0 Hz, 3H, CH ), 0.84 (d, J = 7.2 Hz, 3H, CH ),
CD OD) δ (ppm): 176.2, 171.6, 144.0, 138.7, 135.9, 132.8,
3
3
3
13
.83 (d, J = 6.6 Hz, 3H, CH ); C NMR (150 MHz, CD OD)
130.8 (2C), 128.0 (2C), 127.6, 118.5, 116.2, 76.0, 71.5,
71.5, 61.6, 61.4, 56.4, 52.9 (2C), 52.2 (2C), 48.7, 48.1,
45.6, 43.3, 37.7, 36.8, 35.7, 35.2, 34.6, 30.8, 29.1, 27.2,
26.3, 24.8, 19.5, 17.8, 17.3, 14.0. ESI-HRMS (m/z): calcd.
3
3
δ (ppm): 176.2, 171.6, 163.1, 161.5, 144.0, 138.7, 132.8,
1
7
4
2
31.1, 130.9, 127.6, 118.5, 116.2, 114.6, 114.5, 76.0, 71.5,
1.4, 61.5, 61.4, 56.4, 52.8 (2C), 52.1 (2C), 48.7, 48.1,
5.6, 43.3, 37.7, 36.8, 35.7, 35.2, 34.6, 30.8, 29.1, 27.2,
+
For C H ClN O [M+H] : 726.4249; found, 726.4248.
4
1
61
3
6
6.3, 24.8, 19.5, 17.7, 17.3, 14.0. ESI-HRMS (m/z): calcd.
+
4.1.4.10. 2-[4-(4-bromobenzyl)piperazin-1-yl] ethyl
1R,2R)-2-[(2S,4E,6Z,8R,9S,11R,13S,15S,16S)-7-cyano-
For C H FN O [M+H] : 710.4544; found, 710.4544.
4
1
61
3
6
(
4
(
8
.1.4.8. 2-[4-(2,4-difluorobenzyl) piperazin-1-yl] ethyl
1R,2R)-2-[(2S,4E,6Z,8R,9S,11R,13S,15S,16S)-7-cyano-
,16-dihydroxy-9,11,13,15-tetramethyl-18-
8,16-dihydroxy-9,11,13,15-tetramethyl-18-
oxooxacyclooctadeca-4,6-dien-2-yl]-cyclopentane-1-
carboxylate (4j)

-
1
1
1726, 1636, 1457, 1374, 1275, 1167, 1036, 975 cm ; H
2
0
NMR (600 MHz, CD OD) δ (ppm): 7.20 (d, J = 7.8 Hz, 2H,
3
Ar-H), 7.13 (d, J = 7.8 Hz, 2H, Ar-H), 6.92 (d, J = 11.3 Hz,
1H), 6.60 (dd, J =14.9, 11.3 Hz, 1H), 6.31 (ddd, J = 14.9,
10.1, 4.8 Hz, 1H), 4.96 (dt, J = 10.6, 3.7 Hz, 1H), 4.30 (dt,
J = 11.5, 5.5 Hz, 1H), 4.18 (d, J = 9.8 Hz, 1H), 4.11 (dt, J
= 11.5, 5.5 Hz, 1H), 3.89 (dt, J = 10.3, 3.1 Hz, 1H), 3.53
(d, J = 13.0 Hz, 1H), 3.50 (d, J = 13.0 Hz, 1H), 2.70-2.48
(m, 14H), 2.31 (dd, J = 15.9, 3.0 Hz, 1H), 2.31 (s, 3H, -Ar-
2
9
(
2
1
(
.18 (d, J = 9.8 Hz, 1H), 4.11 (dt, J = 11.5, 5.5 Hz, 1H),
.89 (dt, J = 10.2, 3.1 Hz, 1H), 3.53 (d, J = 13.0 Hz, 1H),
.50 (d, J = 13.0 Hz, 1H), 2.70 (m, 2H), 2.63 (m, 2H), 2.60
CH ), 2.20 (dd, J = 15.9, 10.1 Hz, 1H), 2.00 (m, 2H), 1.83
3
(m, 3H), 1.77 (m, 3H), 1.63 (m, 1H), 1.41 (m, 1H), 1.23
(brt, J = 12.6 Hz, 1H), 1.16 (brt, J = 12.6 Hz, 1H), 1.09
(brt, J = 12.6 Hz, 1H), 0.98 (m, 1H), 0.96 (m, 1H), 0.69
2
2
2
1
H, 1 × -N-CH -), 2.49 (m, 4H, 2 × -N-CH -), 2.31 (dd, J =
5.9, 3.0 Hz, 1H), 2.20 (dd, J = 15.9, 10.2 Hz, 1H), 1.99
(brt, J = 12.6 Hz, 1H), 1.03 (d, J = 6.4 Hz, 3H, CH ), 0.85
2
2
3
(d, J = 6.6 Hz, 3H, CH ), 0.84 (d, J = 7.2 Hz, 3H, CH ),
3
3
13
(
(
m, 2H), 1.83 (m, 3H), 1.77 (m, 3H), 1.65 (m, 1H), 1.41
m, 1H), 1.24 (brt, J = 12.6 Hz, 1H), 1.16 (brt, J = 12.6 Hz,
0.83 (d, J = 6.6 Hz, 3H, CH ); C NMR (150 MHz, CD OD)
3
3
δ (ppm): 176.2, 171.6, 144.0, 138.7, 136.9, 129.4 (2C),
128.5 (2C), 127.6, 118.5, 116.2, 76.0, 71.5 (2C), 62.2,
61.4, 56.4, 52.7 (2C), 52.2 (2C), 48.7, 48.1, 45.6, 43.3,
37.7, 36.9, 35.7, 35.2, 34.6, 30.9, 29.1, 27.2, 26.3, 24.8,
19.8, 19.5, 17.7, 17.3, 14.0. ESI-HRMS (m/z): calcd. For
1
1
H), 1.08 (brt, J = 12.6 Hz, 1H), 0.98 (m, 1H), 0.96 (m,
H), 0.69 (brt, J = 12.6 Hz, 1H), 1.03 (d, J = 6.4 Hz, 3H,
CH ), 0.85 (d, J = 6.0 Hz, 3H, CH ), 0.84 (d, J = 6.0 Hz, 3H,
3
3
1
3
CH ), 0.82 (d, J = 6.0 Hz, 3H, CH ); C NMR (150 MHz,
CD OD) δ (ppm): 176.2, 171.6, 144.0, 138.7, 136.4, 131.1
3
3
+
C H N O [M+H] : 706.4795; found, 706.4797.
3
42 64
3
6
2C), 131.0 (2C), 127.6, 120.7, 118.5, 116.2, 76.0, 71.5,
1.5, 61.6, 61.4, 56.4, 52.9 (2C), 52.2 (2C), 48.7, 48.1,
5.6, 43.3, 37.7, 36.8, 35.7, 35.2, 34.6, 30.8, 29.1, 27.2,
.1.4.13. 2-[4-(naphthalen-2-ylmethyl)piperazin-1-yl] ethyl
1R,2R)-2-[(2S,4E,6Z,8R,9S,11R,13S,15S,16S)-7-cyano-
,16-dihydroxy-9,11,13,15-tetramethyl-18-
6.3, 24.8, 19.5, 17.8, 17.3, 14.0. ESI-HRMS (m/z): calcd.
+
4
1
63
3
6
4
.1.4.11. 2-[4-(4-nitrobenzyl) piperazin-1-yl] ethyl (1R,2R)-
-[(2S,4E,6Z,8R,9S,11R,13S,15S,16S)-7-cyano-8,16-
2
2
0
dihydroxy-9,11,13,15-tetramethyl-18-oxooxacyclooctadeca-
4
,6-dien-2-yl]-cyclopentane-1-carboxylate (4k)
1
Eluent dichloromethane/methanol (80:1). Colorless
2
0
crystal. 90% yield: []
D
-22 (c 0.91, MeOH); UV (MeOH)
λmax (log ε) 256 (4.60) nm; IR v 3430, 2929, 2211, 1727,
636, 1522, 1457, 1346, 1276, 1169, 1030, 973, 857, 743
1
-1
1
cm ; H NMR (600 MHz, CD OD) δ (ppm): 8.20 (d, J = 8.5
3
Hz, 2H, Ar-H), 7.61 (d, J = 8.5 Hz, 2H, Ar-H), 6.93 (d, J =
1
1.3 Hz, 1H), 6.61 (dd, J = 14.9, 11.3 Hz, 1H), 6.32 (ddd,
J = 14.9, 10.1, 4.8 Hz, 1H), 4.96 (dt, J = 10.6, 3.6 Hz, 1H),
Ar), 2.69 (m, 2H), 2.64 (m, 4H, 1 × -N-CH -), 2.57(m, 4H,
2
4
4
1
2
.34 (dt, J = 11.5, 5.5 Hz, 1H), 4.18 (d, J = 9.8 Hz, 1H),
.13 (dt, J = 11.5, 5.5 Hz, 1H), 3.90 (dt, J = 10.3, 3.1 Hz,
H), 3.69 (d, J = 13.0 Hz, 1H), 3.66 (d, J = 13.0 Hz, 1H),
.72-2.49 (m, 14H), 2.32 (dd, J = 15.9, 2.9 Hz, 1H), 2.20
2 × -N-CH -), 2.50 (m, 4H, 2 × -N-CH -), 2.31 (dd, J =
2
2
15.9, 3.0 Hz, 1H), 2.20 (dd, J = 15.9, 10.1 Hz, 1H), 1.97
(m, 2H), 1.82 (m, 4H), 1.75 (m, 2H), 1.63 (m, 1H), 1.39
(m, 1H), 1.23 (m, 1H), 1.15 (brt, J = 12.5 Hz, 1H), 1.06
(brt, J = 12.5 Hz, 1H), 0.96 (m, 2H), 0.69 (brt, J = 12.5 Hz,
(
dd, J = 15.9, 10.2 Hz, 1H), 1.99 (m, 2H), 1.83 (m, 3H),
1
1
0
1
0
.78 (m, 3H), 1.65 (m, 1H), 1.41 (m, 1H), 1.23 (m, 1H),
.16 (brt, J = 12.6 Hz, 1H), 1.08 (brt, J = 12.6 Hz, 1H),
.99 (m, 1H), 0.94 (m, 1H), 0.68 (brt, J = 12.6 Hz, 1H),
.03 (d, J = 6.4 Hz, 3H, CH ), 0.85 (d, J = 6.0 Hz, 3H, CH ),
1H), 1.03 (d, J = 6.6 Hz, 3H, CH ), 0.84 (d, J = 6.6 Hz, 3H,
3
CH ), 0.83 (d, J = 6.6 Hz, 3H, CH ), 0.81 (d, J = 6.6 Hz, 3H,
3
3
13
CH ); C NMR (150 MHz, CD OD) δ (ppm): 176.2, 171.6,
3
3
144.0, 138.7, 134.3, 133.4, 133.0, 128.1, 127.6 (2C),
127.4, 127.3, 127.2, 125.7, 125.5, 118.6, 116.2, 76.0, 71.5
(2C), 62.6, 61.4, 56.4, 52.8 (2C), 52.3 (2C), 48.7, 48.1,
45.6, 43.3, 37.7, 36.9, 35.7, 35.2, 34.6, 30.8, 29.1, 27.2,
26.3, 24.8, 19.5, 17.7, 17.3, 14.0. ESI-HRMS (m/z): calcd.
3
3
.84 (d, J = 6.0 Hz, 3H, CH ), 0.81 (d, J = 6.6 Hz, 3H, CH );
3
3
1
3
C NMR (150 MHz, CD OD) δ (ppm): 176.1, 171.6, 147.3,
3
1
1
45.5, 144.0, 138.8, 129.8 (2C), 127.6, 123.0 (2C), 118.6,
16.2, 76.0, 71.5, 71.5, 61.4, 61.2, 56.4, 53.0 (2C), 52.3
+
(
2C), 48.8, 48.1, 45.6, 43.3, 37.7, 36.8, 35.7, 35.2, 34.6,
forC H N O [M+H] : 742.4795; found, 742.4795.
45
64
3
6
3
0.8, 29.1, 27.2, 26.3, 24.8, 19.5, 17.8, 17.3, 14.0. ESI-
+
4.1.4.14. 2-{4-[2-(1,3-dioxolan-2-yl)ethyl]piperazin-1-yl}
ethyl (1R,2R)-2-[(2S,4E,6Z,8R,9S,11R,13S,15S,16S)-7-
cyano-8,16-dihydroxy-9,11,13,15-tetramethyl-18-
oxooxacyclooctadeca-4,6-dien-2-yl]cyclopentane-1-
carboxylate (4n)
HRMS (m/z): calcd. For C H N O [M+H] : 737.4489;
found, 737.4492.
4
1
61
4
8
4
.1.4.12. 2-[4-(4-methylbenzyl)piperazin-1-yl] ethyl
(1R,2R)-2-[(2S,4E,6Z,8R,9S,11R,13S,15S,16S)-7-cyano-
8
,16-dihydroxy-9,11,13,15-tetramethyl-18-
oxooxacyclooctadeca-4,6-dien-2-yl]-cyclopentane-1-
carboxylate (4l)
2
0
2212, 1729, 1638, 1538, 1460, 1376, 1282, 1163, 1037,
2
0
-1
1
973, 815, 779, 726, 657 cm ; H NMR (600 MHz, CD OD) δ
3
λmax (log ε) 256 (4.40) nm; IR v 3395, 2923, 2862, 2211,
(ppm): 6.83 (d, J = 11.3 Hz, 1H), 6.51 (dd, J = 14.9, 11.3

Hz, 1H), 6.23 (ddd, J = 14.9, 10.1, 4.8 Hz, 1H), 4.86 (dt, J
10.6, 3.7 Hz, 1H), 4.79 (t, J = 4.8 Hz, 1H, -CH), 4.21 (dt,
J = 11.5, 5.5 Hz, 1H), 4.09 (d, J = 9.8 Hz, 1H), 4.04 (dt, J
11.5, 5.5 Hz, 1H), 3.84 (m, 2H), 3.80 (dt, J = 10.0, 3.2
1 × -N-CH -), 2.56 (m, 4H, 2 × -N-CH -), 2.51 (m, 4H, 2 ×
2 2
=
-N-CH -), 2.34 (m, 2H, -CH -cyclopropane), 2.33 (m, 1H),
2
2
2.32 (dd, J = 15.9, 3.0 Hz, 1H), 2.20 (dd, J = 15.9, 10.2
Hz, 1H), 2.00 (m, 2H), 1.83 (m, 3H), 1.78 (m, 3H), 1.65
(m, 1H), 1.43 (m, 1H), 1.23 (m, 1H), 1.17 (brt, J = 12.5 Hz,
1H), 1.09 (brt, J = 12.5 Hz, 1H), 0.99 (m, 1H), 0.96 (m,
1H), 0.69 (brt, J = 12.5 Hz, 1H), 0.56 (m, 2H), 0.18 (m,
=
Hz, 1H), 3.73 (m, 2H), 2.61 (m, 2H), 2.55 (m, 3H, 1 × -N-
CH -), 2.49 (m, 4H, 2 × -N-CH -), 2.44 (m, 3H, 1 × -N-
2
2
CH -), 2.42 (m, 4H, 2 × -N-CH -), 2.23 (dd, J = 15.9, 3.1
2
2
Hz, 1H), 2.11 (dd, J = 15.9, 10.1 Hz, 1H), 1.91 (m, 2H),
2H), 1.03 (d, J = 6.6 Hz, 3H, CH ), 0.86 (d, J = 6.6 Hz, 3H,
3
13
1
1
1
1
.76 (m, 3H), 1.69 (m, 5H), 1.55 (m, 1H), 1.33 (m, 1H),
.14 (brt, J = 12.5 Hz, 1H), 1.07 (brt, J = 12.5 Hz,
H),1.00 (brt, J = 12.5 Hz, 1H), 0.90 (m, 1H), 0.86 (m,
H), 0.60 (brt, J = 12.5 Hz, 1H), 0.94 (d, J = 6.0 Hz, 3H,
CH ), 0.85 (d, J = 5.4 Hz, 6H, 2 × CH ); C NMR (150 MHz,
3
3
CD OD) δ (ppm): 174.5, 170.0, 142.2, 137.0, 125.9, 116.8,
3
114.5, 74.3, 69.9, 69.8, 61.3, 59.8, 54.7, 50.8 (2C), 50.7
(2C), 47.0, 46.4, 43.9, 41.6, 36.1, 35.3, 34.0, 33.5, 32.9,
29.1, 27.4, 25.6, 24.6, 23.1, 17.8, 16.0, 15.6, 12.3, 5.5,
CH ), 0.77 (d, J = 6.6 Hz, 3H, CH ), 0.76 (d, J = 6.6 Hz, 3H,
3
3
1
3
+
CH ), 0.75 (d, J = 6.6 Hz, 3H, CH ); C NMR (150 MHz,
1.4, 1.4. ESI-HRMS (m/z): calcd. For C H N O [M+H] :
3
3
38 62
3
6
CD OD) δ (ppm): 176.2, 171.6, 143.9, 138.7, 127.6, 118.5,
656.4639; found, 656.4640.
3
1
5
3
1
16.2, 102.7, 76.0, 71.5 (2C), 64.5 (2C), 61.4, 56.4, 52.8,
2.6 (2C), 52.5 (2C), 48.7, 48.1, 45.6, 43.3, 37.7, 36.9,
5.7, 35.2, 34.6, 30.8, 30.5, 29.1, 27.3, 26.3, 24.8, 19.5,
7.7, 17.3, 14.0. ESI-HRMS (m/z): calcd. For C H N O
4
.1.4.17. 2-[4-(2-oxopropyl)piperazin-1-yl] ethyl (1R,2R)-2-
[
(2S,4E,6Z,8R,9S,11R,13S,15S,16S)-7-cyano-8,16-
dihydroxy-9,11,13,15-tetramethyl-18-oxooxacyclooctadeca-
39
64
3
8
+
4,6-dien-2-yl]-cyclopentane-1-carboxylate (4q)
[M+H] : 702.4693; found, 702.4693.
.1.4.15. 2-[4-(cyclohexylmethyl)piperazin-1-yl] ethyl
2
0
2
1
(
,16-dihydroxy-9,11,13,15-tetramethyl-18-
6
3.7 Hz, 1H), 4.32 (ddd, J = 11.5, 6.0, 4.8 Hz, 1H), 4.18 (d,
J = 9.8 Hz, 1H), 4.12 (ddd, J = 11.5, 6.0, 4.8 Hz, 1H), 3.90
(dt, J = 10.1, 3.2 Hz, 1H), 3.37 (d, J = 18.0 Hz, 1H), 3.32
(d, J = 18.0 Hz, 1H), 2.72-2.48 (m, 14H), 2.32 (dd, J =
15.9, 3.1 Hz, 1H), 2.20 (dd, J = 15.9, 10.1 Hz, 1H), 2.11 (s,
3H), 2.00 (m, 2H), 1.87-1.76 (m, 6H), 1.65 (m, 1H), 1.42
(m, 1H), 1.24 (m, 1H), 1.16 (brt, J = 12.5 Hz, 1H), 1.10
(brt, J = 12.5 Hz, 1H), 0.98 (m, 2H), 0.68 (brt, J = 12.5 Hz,
2
0
2
1
6
(
6
3
9
1
2
2
2
1H), 1.03 (d, J = 6.6 Hz, 3H, CH ), 0.85 (d, J = 6.6 Hz, 9H,
3
13
3 × CH ); C NMR (150 MHz, CD OD) δ (ppm): 206.3,
3
3
176.1, 171.6, 144.1, 138.7, 127.7, 118.6, 116.1, 76.0, 71.5,
71.4, 61.5, 61.1, 56.5, 52.7 (2C), 52.5 (2C), 52.4, 48.9,
48.1, 45.6, 43.3, 37.8, 36.7, 35.7, 35.2, 34.6, 30.7, 29.1,
27.2, 26.4, 26.3, 21.8, 19.5, 17.8, 17.3, 14.0. ESI-HRMS
+
(
(m/z): calcd. For C H N O [M+H] : 658.4431; found,
37
60
3
7
1
1
2
H), 0.87 (m, 1H), 0.82 (m, 1H), 0.59 (brt, J = 12.5 Hz,
658.4431.
H), 0.94 (d, J = 6.6 Hz, 3H, CH ), 0.76 (d, J = 7.2 Hz, 6H,
3
13
4.1.4.18. 2-(4-allylpiperazin-1-yl) ethyl (1R,2R)-2-
× CH ), 0.75 (d, J = 6.6 Hz, 3H, CH ); C NMR (150 MHz,
3
3
[
(2S,4E,6Z,8R, 9S,11R,13S,15S,16S)-7-cyano-8,16-
CD OD) δ (ppm): 176.2, 171.6, 143.9, 138.7, 127.6, 118.5,
3
dihydroxy-9,11,13,15-tetra-methyl-18-
oxooxacyclooctadeca-4,6-dien-2-yl]cyclopentane-1-
carboxylate (4r)
1
16.2, 76.0, 71.5 (2C), 65.2, 61.4, 56.5, 52.9 (2C), 52.7
(2C), 48.8, 48.1, 45.6, 43.3, 37.7, 36.8, 35.7, 35.2, 34.6,
3
2
4.6, 31.6, 31.5, 30.8, 29.4, 29.1, 27.2, 26.3 (2C), 25.7,
4.8, 19.5, 17.7, 17.3, 14.0. ESI-HRMS (m/z): calcd. For
Eluent dichloromethane/methanol (40:1). Colorless
crystal. 90% yield: [] -36.4 (c 0.44, MeOH); UV (MeOH)
+
2
0
C H N O [M+H] : 698.5108; found, 698.5106.
41
68
3
6
D
λmax (log ε) 256 (4.31) nm; IR v 3401, 2954, 2922, 2855,
.1.4.16. 2-[4-(cyclopropylmethyl)piperazin-1-yl] ethyl
1R,2R)-2-[(2S,4E,6Z,8R,9S,11R,13S,15S,16S)-7-cyano-
,16-dihydroxy-9,11,13,15-tetramethyl-18-
2
1
212, 1729, 1640, 1597, 1459, 1376, 1267, 1165, 1099,
037, 974, 926, 812, 778, 737 cm ; H NMR (600 MHz,
-1 1
CD OD) δ (ppm): 6.82 (d, J = 11.3 Hz, 1H), 6.51 (dd, J =
3
1
5
4.9, 11.3 Hz, 1H), 6.22 (ddd, J = 14.9, 10.1, 4.8 Hz, 1H),
.78 (ddt, J = 17.0, 10.2, 6.8 Hz, 1H), 5.15 (dd, J = 17.0,
Eluent dichloromethane/methanol (30:1). Colorless
1.8 Hz, 1H), 5.11 (dd, J = 10.2, 1.8 Hz, 1H), 4.87 (dt, J =
10.6, 3.7 Hz, 1H), 4.21 (dt, J = 11.5, 5.5 Hz, 1H), 4.09 (d,
J = 9.8 Hz, 1H), 4.04 (dt, J = 11.5, 5.5 Hz, 1H), 3.80 (dt, J
2
0
crystal. 89% yield: []
D
-34.1 (c 0.82, MeOH); UV (MeOH)
λmax (log ε) 256 (4.82) nm; IR v 3399, 2954, 2923, 2212,
1
7
6
6
3
9
1
728, 1636, 1599, 1458, 1374, 1276, 1166, 1035, 976, 826,
= 10.1, 3.2 Hz, 1H), 2.96 (m, 2H, -CH -alkene), 2.61 (m,
2
-
1
1
45, 677, 637 cm ; H NMR (600 MHz, CD OD) δ (ppm):
2H), 2.56 (m, 2H), 2.51 (m, 2H, 1 × -N-CH -), 2.46 (m, 4H,
3
2
.92 (d, J = 11.3 Hz, 1H), 6.60 (dd, J = 14.9, 11.3 Hz, 1H),
.32 (ddd, J = 14.9, 10.1, 4.8 Hz, 1H), 4.97 (dt, J = 10.6,
.7 Hz, 1H), 4.30 (dt, J = 11.5, 5.5 Hz, 1H), 4.18 (d, J =
.8 Hz, 1H), 4.14 (dt, J = 11.5, 5.5 Hz, 1H), 3.89 (dt, J =
0.0, 3.2 Hz, 1H), 2.71 (m, 2H), 2.66 (m, 2H), 2.62 (m, 2H,
2 × -N-CH -), 2.41 (m, 4H, 2 × -N-CH -), 2.23 (dd, J =
2
2
15.9, 3.1 Hz, 1H), 2.11 (dd, J = 15.9, 10.1 Hz, 1H), 1.90
(m, 2H), 1.73 (m, 3H), 1.69 (m, 3H), 1.56 (m, 1H), 1.33
(m, 1H), 1.14 (m, 1H), 1.07 (brt, J = 12.5 Hz, 1H), 1.00
(brt, J = 12.5 Hz, 1H), 0.90 (m, 1H), 0.86 (m, 1H), 0.60

(
brt, J = 12.5 Hz, 1H), 0.94 (d, J = 6.6 Hz, 3H, CH ), 0.76
4YYE) and compound borrelidin was optimized and saved
for the next docking experiments.
3
(d, J = 6.6 Hz, 6H, 2 × CH ), 0.75 (d, J = 6.6 Hz, 3H, CH );
3
3
13
C NMR (150 MHz, CD OD) δ (ppm): 176.2, 171.6, 143.9,
3
Since the docking studies were performed by Glide in
Schrodinger (version 2014) in subsequent works, the
protein was prepared by Schrodinger's protein preparation
wizard as well and crystallographic water molecules were
deleted as there was no water molecules exist any co-
ordination with the ligand. Other processes of preparation
include correcting protein structure defects, building the
missing side chain atoms and adding hydrogen atoms. Both
the acidic and basic amino acid residues were disposed into
the appropriate ionization states at pH 7.4. Ultimately, the
energy of the protein was minimized till RMSD value of
1
6
4
2
38.7, 133.6, 127.6, 118.5, 118.3, 116.2, 76.0, 71.5, 71.4,
1.5, 61.1, 56.4, 52.6 (2C), 52.3 (2C), 48.7, 48.1, 45.6,
3.3, 37.7, 36.9, 35.7, 35.2, 34.6, 30.8, 29.1, 27.2, 26.3,
4.8, 19.5, 17.7, 17.3, 14.0. ESI-HRMS (m/z): calcd. For
+
C H N O [M+H] : 642.4482; found, 642.4484.
37
60
3
6
4
.2. Biological Assays
4.2.1. In vitro Antifungal Activity Evaluation by MIC Assay
The MIC values of borrelidin derivatives were
43
determined by using a micro broth dilution method. C.
albicans (ATCC MYA-2876) and C. parapsilosis (ATCC
0
.30 Å was done using OPLS_2005 force field to relieve the
steric clashes among the residues due to addition of
hydrogen atoms. Next, the generated grid file from the
prepared protein was used for Glide XP docking
2
2019) were obtained from China General Microbiological
Culture Collection Center. A. fumigatus (CCTCC AF 93048)
was obtained from China Center for Type Culture Collection.
Antifungal assay were performed in RPMI-1640 broth (10.4
calculations. The minimized conjugate gradient output of
the reference ligand. XP Glide scoring function was used to
order the best ranked compounds and the specific
interactions like hydrogen bonding and pi-pi stacking were
analyzed by formed 2D interactions diagram using in Glide
module. Finally, the docked completed file was imported
into software Discovery Studio 3.0 and PyMOL to
g/L RPMI-1640, 2 g/L NaHCO , 34.53 g/L MOPs, pH 7.0).
3
The tested compounds were dissolved in DMSO and 2-fold
serially diluted to eight different concentrations (10-0.31
mg/mL), and the above samples (1 μL) was added to each
well. Afterwards, prepared fungal suspensions containing 2
3
×
10 cfu/mL for C. albicans, C. parapsilosis and A.
investigate the 3D interaction relationship and the
conformation of the compound on the protein surface.
fumigatus were added to each well of 96-well microtiter
plates, and the plates were incubated for 48 h at 28 °C for
C. albicans, C. parapsilosis and A. fumigatus. The minimum
concentration of sample showing no turbidity was recorded
as the MIC.
Acknowledgments
This work was funded by National Natural Science Foundation
of China (No. 81573327) and the Fundamental Research Funds
for the Central Universities, China (Nos. N172004004 and
N172008008).
4.2.2. Cytotoxicity Assay
The human kidney epithelial cell line (293T) were
obtained from National Infrastructure of Cell Line Resource
Beijing, China), and cultured in DMEM supplemented with
0% fetal bovine serum (FBS), 1% L-glutamine, 100
(
1
References and notes
units/mL penicillin and 100 μg/mL streptomycin at 37 °C in
a humidified 5% CO atmosphere incubator (Thermo Fisher
1. Leeflang, M.M.; Debetsossenkopp, Y.J.; Visser, C.E.
Database. Syst. Rev. 2008, 4, CD007394.
2. De, P.G.; Tumbarello, M. Curr. Opin. Crit. Care. 2015, 21,
421-429.
2
Scientific, Waltham, MA, USA). The cytotoxicity and
proliferation inhibition were quantified by the MTT assay.
3
2
93T (7 × 10 cells/well) was seeded in 96-well plates
3. Turgut, M.; Ozsunar, Y.; Oncü, S. Surg. Neurol. 2008, 69,
169-174.
4. Wong, S.S.; Kao, R.Y.; Yuen, K.Y. Plos One. 2014, 9,
e85836.
5. Bitar, I.; Khalaf, R.A.; Harastani, H. Biomed. Res. Int. 2014,
2014, 931372.
6. Lee, J.; Choi, Y.; Woo, E.R.; Lee, D.G. Biochem. Biophys.
Res. Commun. 2009, 379, 676-680.
7. Berger, J.; Jampolsky, L.M.; Goldberg, M.W. Arch.
Biochem. 1949, 22, 476-478.
IC50 values were calculated by a non-linear regression
analysis using SPSS 13.0 (SPSS Inc., Chicago, IL, USA).
8.
Keller-Schierlein, W. Helv. Chim. Acta. 1967, 50, 731-753.
Kuo, M.S.; Yurek, D.A.; Kloosterman, D.A. J. Antibiot.
9
1
1
1
1
1
.
1
989, 42, 1006-1007.
4.2.3. Molecular docking study
0. Anderson, B.F., Herlt, A.J., Rickards, R.W., Robertson, G.B.
Aust. J. Chem. 1989, 42, 717-730.
1. Hanessian, S.; Yang, Y.; Giroux, S.; Mascitti, V.; Ma, J.;
Raeppel, F. J. Am. Chem. Soc. 2003, 125, 13784-13792.
2. Duffey, M.O.; LeTiran, A.; Morken, J.P. J. Am. Chem. Soc.
Download the homo threonyl-tRNA synthetase with co-
crystal borrelidin (PDB: 4P3N) and threonyl-tRNA
synthetase of the yeast mitochondrial (PDB: 4YYE) from
the Protein Data Bank (http://www.rcsb.org/pdb). Align
their structures to find out the binding site by using
Discovery Studio 3.0 software. This software was also
applied as for the preparation of protein, during which
missing amino acid residues complemented, hydrogen
atoms added, all water molecules removed and missing
side chains fixed. The complex of protein structure (PDB:
2
003, 125, 1458-1459.
3. Vong, B.G.; Kim, S.H.; Abraham, S.; Theodorakis, E.A.
Angew. Chem. 2004, 43, 3947-3951.
4. Nagamitsu, T.; Takano, D.; Fukuda, T.; Otoguro, K.;
Kuwajima, I.; Harigaya, Y.; Omura, S. Org. Lett. 2004, 6,
1
865-1867.

1
1
5. Olano, C.; Wilkinson, B.; Moss, S.J.; Brana A.F.; Mendez,
C.; Leadlay P.F.; Salas, J.A. Chem. Commun. 2003, 22,
32. Ma, X.Y.; He, Y.; Hu, Y.L.; Lu, M. Tetrahedron Lett. 2012,
53, 449-452.
33. Taylor, R.D.; MacCoss, M.; Lawson, A.D. J. Med. Chem.
2014, 57, 5845-5859.
34. Hemmerling, F.; Hahn, F. Beilstein. J. Org. Chem. 2016, 12,
1512-1550.
35. Padmavathi, V.; Venkata Subbaiah, D.R.; Mahesh, K.;
Radha Lakshmi, T. Chem. Pharm. Bull. 2007, 55, 1704-
1709.
36. Bazgir, A.; Khanaposhtani, M.M.; Soorki, A.A. Bioorg.
Med. Chem. Lett. 2008, 18, 5800-5803.
37. Patel, R.V.; Park, S.W. Mini. Rev. Med. Chem. 2013, 13,
1579-1601.
2
780-2782.
6. Olano, C.; Wilkinson, B.; Sanchez, C.; Moss, S.J.; Sheridan,
R.; Math, V.; Weston, A.J.; Brana, A.F.; Martin, C.J.;
Oliynyk, M.; Mendez, C.; Leadlay, P.F.; Salas, J.A. Chem.
Biol. 2004, 11, 87-97.
7. Olano, C.; Moss, S.J.; Brana, A.F.; Sheridan, R.M.; Math,
V.; Weston, A.J.; Mendez, C.; Leadlay, P.F.; Wilkinson, B.;
Salas, J.A. Mol. Microbiol. 2004, 52, 1745-1756.
8. Moss, S.J.; Carletti, I.; Olano, C.; Sheridan, R.M.; Ward, M.;
Math, V.; Nur, E.A.M.; Brana, A.F.; Zhang, M.Q.; Leadlay,
P.F.; Mendez, C.; Salas, J.A.; Wilkinson, B. Chem.
Commun. 2006, 22, 2341-2343.
1
1
3
8. Suresh, N.; Nagesh, H.N.; Renuka, J.; Rajput, V.; Sharma,
R.; Khan, I.A.; Kondapalli Venkata Gowri C.S. Eur. J. Med.
Chem. 2014, 71, 324-332.
1
2
2
9. Lumb, M.; Macey, P.E.; Spyvee, J.; Whitmarsh, J.M.;
Wright, R.D. Nature 1965, 206, 263-265.
0. Singh, S.K.; Gurusiddaiah, S.; Whalen, J.W. Antimicrob.
Agents. Chemother. 1985, 27, 239-245.
39. Friesner, R.A.; Banks, J.L.; Murphy, R.B.; Halgren, T.A.;
Klicic, J.J.; Mainz, D.T.; Repasky, M.P.; Knoll, E.H.;
Shelley, M.; Perry, J.K.; et al. J. Med. Chem. 2004, 47,
1739-1749.
1. Gao, Y.M.; Wang, X.J.; Zhang, J.; Li, M.; Liu, C.X.; An, J.;
Jiang, L.; Xiang, W.S. J. Agric. Food. Chem. 2012, 60,
9
874-9881.
40. Kontoyianni, M.; Sokol, G.S.; McClellan, L.M. J. Comput.
Chem. 2005, 26, 11-22.
41. Sugawara, A.; Tanaka, T.; Hirose, T.; Ishiyama, A.;
Iwatsuki, M.; Takahashi, Y.; Otoguro, K.; Omura, S.;
Sunazuka, T. Bioorg. Med. Chem. Lett. 2013, 23, 2302-
2305.
2
2
2
2
2. Habibi, D.; Ogloff, N.; Jalili, R.B.; Yost, A.; Weng, A.P.;
Ghahary, A.; Ong, C.J. Invest. New. Drugs. 2012, 30, 1361-
1
370.
3. Ruan, B.; Bovee, M.L.; Sacher, M.; Stathopoulos, C.;
Poralla, K.; Francklyn, C.S.; Soll, D. J. Biol. Chem. 2005,
2
80, 571-577.
42. Novoa, E.M.; Camacho, N.; Tor, A.; Wilkinson,B.; Moss,
S.; Marin-Garcia, P.; Azcarate, I.G. Bautista, J.M.; Mirando,
A.C.; Francklyn, C.S.; Varon, S.; Royo, M.; Cortes, A.;
Ribas de Pouplana, L. Proc. Natl. Acad. Sci. USA. 2014,
111, E5508-5517.
4. Ishiyama, A.; Iwatsuki, M.; Namatame, M.; Nishihara-
Tsukashima, A.; Sunazuka, T.; Takahashi Y.; Omura, S.;
Otoguro K. J. Antibiot. 2011, 64, 381-384.
5. Wilkinson, B.; Gregory, M.A.; Moss, S.J.; Carletti, I.;
Sheridan, R.M.; Kaja, A.; Ward, M.; Olano, C.; Mendez, C.;
Salas, J.A.; Leadlay, P.F.; vanGinckel, R.; Zhang, M.Q.
Bioorg. Med. Chem. Lett. 2006, 16, 5814-5817.
43. Zhao, J.; Mou, Y.; Shan, T.; Li, Y.; Zhou, L.; Wang, M.;
Wang, J. Molecules 2010, 15, 7961-7970.
2
2
6. Paetz, W.; Nass, G. Eur. J. Biochem. 1973, 35, 331-337.
7. Gerken, S.C.; Arfin, S.M. J. Biol. Chem. 1984, 259, 9202-
Supplementary Material
1
9
206.
Supplementary materials are available online. The H-NMR,
C-NMR and IR spectra of target compounds.
1
3
2
2
3
8. Nass, G.; Poralla, K. Mol. Gen. Genet. 1976, 147, 39-43.
9. Nass, G.; Hasenbank, R. Mol. Gen. Genet. 1970, 108, 28-32.
0. Fang, P.; Yu, X.; Jeong, S.J.; Mirando, A.; Chen, K.; Chen,
X.; Kim, S.; Francklyn, C.S.; Guo, M. Nat. Commun. 2015,
Click here to remove instruction text...
6
, 6402.
3
1. Gao, X.X.; Jiang, Y.; Han, L.; Chen, X.; Hu, C.J.; Su, H.;
Mu, Y.; Guan, P.P.; Huang X.S. Rsc Adv. 2017, 7, 44401-
4
4409.

Graphical Abstract
Design, Synthesis, and Antifungal Evaluation of Borrelidin Derivatives
Leave this area blank for abstract info.
a
a
b
a,
a
Caijuan Hu , Hao Su , Jinghan Luo , Li Han *, Qingyin Liu ,
a
a
a
b,
a,
Wenxi Wu , Yu Mu , Peipei Guan , Tiemin Sun * and Xueshi Huang *
a
College of Life and Health Sciences, Northeastern University, Shenyang 110819, China;
b
Key Laboratory of Structure-Based Drug Design and Discovery, Shenyang Pharmaceutical University, Shenyang 110016, China

Highlights





Synthesis and antifungal evaluation of forty-
seven borrelidin analogs.
Fragment -OCH CH N- was essential for
2
2
antifungal activity.
3b exhibited activity against C. albicans which
was insensitive to borrelidin.
Compared with borrelidin, 4n and 4r presented
extra activity against A. fumigatus.
Carboxyl substituents in borrelidin existed
extra interaction with fungal ThrRS.