Synthesis and biological evaluation of oleanolic acid derivatives as novel inhibitors of protein tyrosine phosphatase 1B

Article abstract of DOI:10.3987/COM-12-12445

A series of oleanolic acid (OA) derivatives have been synthesized and their inhibitory effects on PTP1B, TCPTP and related PTPs are evaluated. Some compounds with five-membered heterocyclic ring-fused at C-2, C-3 positions showed a dramatic increase in in

Full text of DOI:10.3987/COM-12-12445

HETEROCYCLES, Vol. 85, No. 5, 2012
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HETEROCYCLES, Vol. 85, No. 5, 2012, pp. 1117 - 1139. © 2012 The Japan Institute of Heterocyclic Chemistry
Received, 10th February, 2012, Accepted, 26th March, 2012, Published online, 2nd April, 2012
DOI: 10.3987/COM-12-12445
SYNTHESIS AND BIOLOGICAL EVALUATION OF OLEANOLIC ACID
DERIVATIVES AS NOVEL INHIBITORS OF PROTEIN TYROSINE
PHOSPHATASE 1B
Hui Li,a Hui Zou,b Lixin Gao,c Ting Liu,a Fan Yang,a Jingya Li,c Jia Li,*c
Wen-Wei Qiu,*band Jie Tanga
a. Shanghai Engineering Research Center of Molecular Therapeutics and New
Drug Development, East China Normal University, Shanghai 200062, P. R. China
b. Department of Chemistry, East China Normal University, Shanghai 200062, P.
R. China. (phone: +86-21-62237950; fax: +86-21-62237950; e-mail:
wwqiu@chem.ecnu.edu.cn)
c. National Center for Drug Screening, Shanghai Institute of Materia Medica,
Shanghai Institute for Biological Science, Chinese Academy of Science, Shanghai
201203, P. R. China. (phone: +86-21-50801313×132; fax: +86-21-50801552;
e-mail: jli@mail.shcnc.ac.cn)
Abstract – A series of oleanolic acid (OA) derivatives have been synthesized and
their inhibitory effects on PTP1B, TCPTP and related PTPs are evaluated. Some
compounds with five-membered heterocyclic ring-fused at C-2, C-3 positions
showed a dramatic increase in inhibition, the two most potent PTP1B inhibitors
19 (IC50 = 0.91 μM) and 21 (IC50 = 0.98 μM) showed about 3-fold more potent
than lead compound OA. Some C-ring modified OA analogs showed high
selectivity for PTP1B over TCPTP, among them, 50 possessed the best selectivity
of 6.6-fold.
INTRODUCTION
Protein tyrosine phosphatases (PTPs) are expressed in insulin-sensitive tissues, which can function as
negative modulators in insulin signal transduction by dephosphorylation of tyrosyl residues.1–4 Protein
tyrosine phosphatase 1B (PTP1B), a prototypical member of the protein tyrosine phosphate superfamily,
is a key negative regulator of both insulin and leptin signaling pathway by dephosphorylating the insulin
receptor (IR),5 insulin receptor substrates (IRS)6 and Janus kinase 2 (JAK2).7,8 Previous studies have

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HETEROCYCLES, Vol. 85, No. 5, 2012
reported that PTP1B knockout mice not only display increased insulin sensitivity in liver and muscle
tissues but also are resistant to high fat diet-induced obesity.7,9-11 PTP1B antisense oligonucleotide
treatment can modulate obesity related fat storage and lipogenesis in adipose, and improve insulin
sensitivity in animal models of type 2 diabetes.12,13 These investigations suggest that inhibition of
PTP1B may anti-type 2 diabetes by increasing insulin sensitivity9 and resistance in obesity.10 In recent
years, PTP1B inhibitors are regarded as agents in the treatment of type 2 diabetes and obesity.14,15
A
series of synthetic inhibitors with submicro-, even nano-molar activity were discovered, however, few
was further development to clinical trials, mainly for two reasons. Firstly, the poor bioavailability is an
important issue, because of most active site-directed inhibitors reported to date possess a high charge
density by mimicking the phosphate group in IRS.16 Secondly, the low selectivity between PTP1B and
the most homogeneous T-cell protein tyrosine phosphatase (TCPTP) is another important issue, because
of all PTPs share a high degree of structural conservation in the active site.17
Natural products play a major role in drug discovery, about 60% of anticancer and 75% of anti-infective
drugs approved from 1981-2002 could be traced to natural origins.18 In searching for novel PTP1B
inhibitors that derived from natural products, researchers have found trodusquemine (is currently
concducting a phase I clinical challenge by Genaera19) and pentacyclic triterpenoids,20,21 including
oleanolic acid (OA) and its derivatives.22 It is well-known that pentacyclic triterpenes are too
hydrophobic to have acceptable water solubility and related pharmacokinetic properties. Initial SAR
studies for analogs of OA with modified various substituents on C-3 and C-28 positions indicated a
strong preference for the hydrophobic group and these derivatives had no obvious selectivity for PTP1B
over TCPTP (with 77% sequences identity with PTP1B). Our previous study was focused on improving
water solubility, increasing inhibitory activity and selectivity between the two homogeneous enzymes by
introducing a series of heterocyclic rings at C-2 and C-3 positions of OA.23 According to our previous
study, the inhibition of the five-membered heterocyclic ring-fused OA derivatives is more potent than
the six-membered analogs. Therefore, we herein reported a subsequent work of our previous research.
We further introduced five-membered hydrophilic heterocyclic rings at C-2 and C-3 positions and
modified the C-ring of OA respectively. All of these OA derivatives were evaluated in the enzyme
inhibition assay against PTP1B and TCPTP, and selected compounds of the series were evaluated in
related PTPs, such as SHP-1, SHP-2 and LAR.
RESULTS AND DISCUSSION
1. Chemistry. Benzylation of OA, followed by IBX oxidation to give 2. Reaction of 2 with isoamyl
nitrite in the presence of t-BuOK in t-BuOH furnished 3.24 Compound 3 was treated with hydroxylamine

HETEROCYCLES, Vol. 85, No. 5, 2012
1119
hydrochloride in refluxing pyridine to give 4. Cyclization of 4 was performed by heating in the presence
of KOH to give 5.25 Compound 6 was produced by debenzylation of 5 over palladium/carbon in MeOH
with high yield. Reaction of 3 with zinc powder in AcOH and Ac2O produced 7. Compound 8 was
afforded by reaction of 7 with phosphorus oxychloride in dry pyridine under nitrogen atmosphere.26
Compound 9 was obtained by debenzylation of 8 with the same procedure as preparation of 6, as shown
in scheme 1.
C
CO2Bn
CO2Bn
c
CO2H
b
a
2
3
HO
O
HO
2
1
OA
CO2Bn
CO2Bn
e
CO2R1
HON
O
d
HON
HON
N
N
O
3
4
5
g
R1 = Bn
f
6 R1 = H
CO2R1
H
N
CO2Bn
h
N
O
O
O
8
7
R1 = Bn
f
9 R1 = H
Scheme 1
a) BnBr, K2CO3, DMF, rt, 12 h, 92%. b) IBX, DMSO, THF, rt, 6 h, 85%. c) isoamyl nitrite, t-BuOK,
t-BuOH, rt, 5 h, 86%. d) hydroxylamine hydrochloride, pyridine, reflux, 5 h, 75%. e) KOH, ethylene
glycol, dioxane, reflux, 5 h, 60%. f) 10% Pd/C, H2, MeOH, rt, 12 h, 92% for 6, 82% for 9. g) Zn dust,
AcOH, Ac2O, rt, 12 h, 86%, h) POCl3, pyridine, rt, 12 h, 75%.
Compound 10 was produced by Oxidation of OA with IBX. Treatment of 10 with various acylation
reagents afforded acylated intermediates 11, 12, 13, 14. Compound 15 was afforded by treatment of 11
with hydroxylamine hydrochloride in refluxing pyridine. Reaction of 12 with hydrazine hydrate in acetic
acid produced 16. Compounds 17 and 18 were obtained by treatment of 13 with hydrazine hydrate or
hydroxylamine hydrochloride respectively.25 Compounds 19 and 21 were obtained by cyclization of 14
with the same procedure as preparation of 17 and 18. Compounds 20 and 22 were synthesized by

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HETEROCYCLES, Vol. 85, No. 5, 2012
hydrolysis of 19 and 21 in the presence of NaOH in THF/MeOH, as shown in scheme 2.
OH
CO2H
CO2H
CO2H
a
b
c
OA
O
N
O
O
15
10
11
b
F3C
HN
CO2H
O
O
CO2H
N
R
16
d
R = CF3
12
O
f
CO2H
R = OEt
13
14
e
O
R = CO2Et
N
H
g
18
d
O
CO2H
HN
N
RO2C
H
CO2H
RO2C
HN
CO2H
17
N
O
N
21
22
R = Et
R = H
R = Et
19
h
h
20 R = H
Scheme 2
a) IBX, DMSO, THF, rt, 5 h, 90%. b) RCO2Et (R = CF3, OEt, CO2Et), NaH, THF, rt, 12 h or HCO2Et,
MeONa, toluene, rt, 1 h, 70% for 11, 78% for 12, 76% for 13, 78% for 14. c) hydroxylamine
hydrochloride, pyridine, reflux, 5 h, 74%. d) 85% hydrazine hydrate, AcOH, reflux, 5 h, 81%. e) 85%
hydrazine hydrate, EtOH, reflux, 5 h, 67%. f) hydroxylamine hydrochloride, AcONa, EtOH, reflux, 24 h,
68%. g) hydroxylamine hydrochloride, EtOH, reflux, 5 h. h) 2 M NaOH, THF, MeOH, reflux, 5 h.
Acetylation of 1 followed by oxidation with PDC produced 24.27 Compound 25 was generated by
removal of acetyl group of 24 in the presence of NaOH in MeOH/THF and benzyl group of 24 in the
presence of Pd/C respectively. The intermediate 26 was synthesized by reaction of 23 with mCPBA.28
Compound 27 was obtained by removal of acetyl group and benzyl group of 26 with the same
procedure as preparation of 25. Compound 29 was afforded by dehydrogenation of 26 in the presence of
Br2 and HBr in HOAc, then removal of acetyl and benzyl group. Compound 26 reacted with
hydroxylamine hydrochloride in pyridine to produce 30. Compound 31 was prepared by deprotection of

HETEROCYCLES, Vol. 85, No. 5, 2012
1121
acetyl group and benzyl group of 30. Compound 33 was generated by Beckmann rearrangement of 30,
then deprotection of acetyl group and benzyl group, as shown in scheme 3.
O
O
CO2H
CO2Bn
O
c
CO2H
HO
AcO
24
25
HO
b
27
c
O
O
CO2Bn
d
CO2Bn
CO2Bn
a
e
1
AcO
AcO
AcO
28
c
23
26
f
O
NOH
O
NH
CO2Bn
CO2H
CO2Bn
g
AcO
HO
AcO
29
32
30
c
c
O
NOH
NH
CO2H
CO2H
HO
HO
31
33
Scheme 3
a) Ac2O, DMAP, pyridine, rt, 12 h, 90%. b) pyridinium dichromate, DCM, rt, 3 h, 50%. c) 1. 2 M NaOH,
THF, MeOH, rt, 12 h; 2. 10% Pd/C, MeOH, rt, 12 h, 68-80% (over two steps). d) mCPBA, CHCl3, rt, 24
h, 60%. e) HBr, Br2, AcOH, 60 oC, 78%. f) hydroxylamine hydrochloride, pyridine, reflux, 5 h. g) SOCl2,
THF, rt, 12 h, 86%.
Compound 1 was protected by TBS group and then reacted with mCPBA to produce 35. Reduction of
carbonyl goup of 35 with NaBH4 yielded 36 and 37; the ratio is 2/3. We also found although 37 could be
methylation or acylation, 36 was very difficult to take place these reaction. We speculated that 36
possessed strong steric hindrance. Compounds 40 and 41 were obtained by deprotection of TBS and
benzyl group of 36 and 37. The structure and configuration of 39 were proven by single-crystal X-ray

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HETEROCYCLES, Vol. 85, No. 5, 2012
diffraction analysis29 (Figure 1). Compound 43 was synthesized by methylation of 37, then removal of
TBS and benzyl group of 37. Acylation of 37 with benzoyl chloride or chloroacetyl chloride afforded 44
and 45. Compound 46 was produced by removal of TBS and benzyl group of 44. Deprotection of TBS
group of 45 afforded 47. Compounds 50 and 51 were generated by reaction of 47 with piperidine or
morpholine, then removal of benzyl group, as shown in scheme 4.
O
R
CO2Bn
b
CO2Bn
c
CO2Bn
a
d
1
TBSO
TBSO
TBSO
35
34
36 R = OH (a)
37 R = OH (b)
R1
g
f
CO2R'2
R2
O
O
HO
CO2Bn
CO2R2
38 R1 = OH (a) R'2 = Bn
R' = Bn
39 R1 = OH (b)
40
2
e
R1 = OH (a) R'2 = H
R1
41 R1 = OH (b) R'2 = H
42 R1 = OTBS, R2 = Bn
43 R1 = OH, R2 = H
R1 = OTBS, R2 = COPh
44
d, e
d, e
45 R1 = OTBS, R2 = COCH2Cl
47 R1 = OH, R2 = COCH2Cl
d
PhO2C
h
CO2H
R2H2CCO2
HO
CO2R1
46
48 R1 = Bn R2 =
N
N
HO
49
50
R2 =
O
O
R1 = Bn
R1 = H
O
e
CO2H
N
N
R2 =
R2 =
O
51 R1 = H
O
52
HO2C
O
Scheme 4
a) TBSCl, imidazole, DMF, 60 oC, 5 h, 83%. b) mCPBA, CHCl3, rt, 12 h, 67%. c) NaBH4, THF/H2O, rt,
5 h, 89%, (36/37 = 2:3). d) BF3·Et2O, CHCl3, 0 oC, 5 h, 82%. e) Pd/C, H2, MeOH, rt, 12 h, 78% for 40,
75% for 41, 74% for 43 and 75% for 46 over two steps. f) MeI, NaH, DMF, rt, 3 h, 85%. g) benzoyl
chloride or chloroacetyl chloride, pyridine, DCM, rt, 5 h, 78% for 44 and 90% for 45. h) piperidine or
morpholine, DCM, reflux, 6 h, 85% for 48 and 49.

HETEROCYCLES, Vol. 85, No. 5, 2012
1123
Figure 1. Single-crystal X-ray diffraction analysis of 39
2. In vitro biological evaluation. In this paper, we further synthesized OA derivatives based on our
previous work.23 All these synthetic derivatives were evaluated in the enzyme inhibition assay against
PTP1B by the method of p-nitrophenyl phosphate using compound 52 as a reference compound.30
Homogeneous TCPTP inhibitory activities were investigated simultaneously by the same method for
further selectivity studying (Tables 1 and 2). We also tested the inhibitory activity of some synthetic
derivatives on several other critical PTPs, which negatively regulate insulin dephosphorylation, such as
leukocyte antigen-related phosphatase (LAR), src homology phosphatase-1 (SHP-1) and src homology
phosphatase-2 (SHP-2) (Table 3).
The IC50 of OA derivatives with heterocyclic ring-fused at C-2, C-3 positions (6-22) were tested on
PTP1B and TCPTP. The results (Table 1) indicated that we obtained two most potent inhibitors 19 (IC50
= 0.91 M) and 21 (IC50 = 0.98M) which showed about 3-fold more potent than lead compound OA
on PTP1B. The inhibitory activity was decreased by introducing hydrophilic substituents at the
five-membered heterocycles, such as carbonyl group (17 and 18), especially the formyloxy group (20
and 22), and increased by introducing hydrophobic substituents, such as trifluoromethyl group (16),
especially the ethyl formyloxy group (19 and 21). The selectivity between PTP1B and TCPTP of these
heterocyclic derivatives was not significantly improved, among them 16 possessed the best selectivity of
4.4-fold.

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HETEROCYCLES, Vol. 85, No. 5, 2012
Table 1. Inhibitory activity of OA derivatives with heterocyclic ring-fused at C-2, C-3 positions against
PTP1B and TCPTP
IC50 (M)
TCPTP/PTP1Ba
Compounds
PTP1B
TCPTP
7.78
_
_
+
1.55
+
2.96 0.26
2.6
3.5
2.3
2.6
4.4
OA
6
9
_
_
+
+
2.71
0.35
9.58 1.64
_
+
_
+
2.86 0.34
6.59 1.10
_
_
+
+
3.05 0.23
15
16
17
7.99 1.29
_
+
_
+
6.15 1.40
1.39 0.27
_
_
+
+
6.09 0.58
2.99 0.44
2.0
2.0
_
_
+
+
18
19
7.89 0.95
3.89 0.63
_
_
+
+
2.01 0.34
0.91 0.13
2.2
2.7
2.5
_
_
+
+
20
21
22
52b
23.8 7.12
8.87 1.55
_
+
_
+
2.46 0.22
0.98 0.14
_
+
_
+
7.93 3.69
6.54 0.68
1.2
1.6
_
+
_
+
6.24 0.19
3.89 0.08
a) TCPTP/PTP1B, the ratio of IC50 of TCPTP and PTP1B. b) Positive control.
The PTP1B inhibition of the C-ring modified derivatives (25-51) was decreased dramatically, with the
exception of 46. The ring expansion compound 33 was obtained by Beckman rearrangement of 31,
which PTP1B inhibitory potency was decreased at least 2-fold. The stereo configuration of C-12
hydroxyl is important since the activity of -hydroxyl compound (41) was about half of -hydroxyl
compound (40). Compared with 41, the C-12 -hydroxyl substituted with hydrophobic groups, such as
methyl, benzoyl, piperidine N-acetyl and morpholine N-acetyl group (43, 46, 50 and 51) could
ameliorate the PTP1B inhibitory potency. Although the PTP1B inhibitory potency of these C-ring
modified derivatives was decreased dramatically, the selectivity between PTP1B and TCPTP of some
derivatives was improved significantly, especially 25, 50 and 51 possessed more than 6-fold selectivity.
Table 2. Inhibitory activity of C-ring modified OA derivatives against PTP1B and TCPTP
a) Not determined. b) TCPTP/PTP1B, the ratio of IC50 of TCPTP and PTP1B. c) Positive control.

HETEROCYCLES, Vol. 85, No. 5, 2012
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Moreover, some derivatives (16, 19, 21, 25, 50 and 51), which had potent inhibition or high selectivity,
were also evaluated on homogenous enzymes LAR, SHP-1 and SHP-2 (Table 3). The results showed
that these compounds had no obvious inhibition against LAR, SHP-1 and SHP-2. Compared to OA,
these tested derivatives developed by our group clearly have better selectivity between PTP1B and
SHP-1.
Table3. Inhibitory activity of selected OA derivatives against related PTPs
IC50 (M)
Compounds
SHP-1
SHP-2
> 40
LAR
> 40
OA
16
19
21
_
+
5.39
31.3
> 40
> 40
> 40
> 40
> 40
> 40
> 40
> 40
> 40
25
50
51
> 40
> 40
> 40
> 40
> 40
> 40
> 40
> 40
> 40
CONCLUSION
In summary, we synthesized a series of novel OA derivatives with five-membered heterocyclic
ring-fused at C-2, C-3 positions and the C-ring modified OA analogs efficiently. The five-membered
heterocyclic ring-fused compounds had improved inhibitory activity on PTP1B, and the most potent
compounds possessed 3-fold potency compare to OA; however, the selectivity between PTP1B and
TCPTP was not obviously improved. The C-ring modified compounds had improved selectivity for
PTP1B over TCPTP, some compounds possessed up to 6-fold selectivity; however, the inhibitory
potency on PTP1B was decreased dramatically. We hope that merge the two parts of works,
modification of the C-2, C-3 positions and the C-ring simultaneously, would afford the potent and high
selectivity PTP1B inhibitors.
EXPERIMENTAL
1
General. H (400 and 500 MHz) and 13C (100 and 125 MHz) NMR spectra were recorded on a
JEOL-400 or Bruker AM-500 Fourier transform spectrometer. The chemical shifts were reported (in
ppm) using the= 7.26, 2.5 signals of CDCl3, DMSO-d6 (1H NMR), and using the= 77.23, 39.51
signals of CDCl3, DMSO-d6 (13C NMR) as internal standards. High-resolution mass data were obtained
on a Micromass Tof II spectrometer.
General procedure A, oxidation of 3-OH group of OA derivatives. OA derivative (1 mmol) was
dissolved in DMSO (30 mL). To this soln., IBX (0.56 g, 2 mmol) was added. After stirring for 12 h at rt,

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HETEROCYCLES, Vol. 85, No. 5, 2012
the reaction mixture was poured into H2O (30 mL), the precipitate was filtered, and the filtrate was
extracted with AcOEt (3×20 mL). The combined organic layer was washed with brine, dried over
anhydrous Na2SO4 and concentrated under reduced pressure. The residue was purified by column
chromatography (CC).
General procedure B, acylation of C-2 position of 10. OA derivative (0.2 mmol) was dissolved in dry
THF (20 mL). To this soln., NaH (40 mg, 1 mmol, 60% in oil) was added. After stirring for 1 h at rt, the
acylation reagent RCO2Et (R = CF3, OEt, CO2Et) (1 mmol) was added. After stirring for 12 h at rt, the
reaction mixture was poured into H2O (40 mL), the pH was adjusted to 6-7 by adding 5% HCl, and the
mixture was extracted with AcOEt (3×20 mL). The combined organic layer was washed with brine,
dried over anhydrous Na2SO4 and concentrated under reduced pressure. The residue was purified by CC.
General procedure C, deprotection of benzyl group of OA derivatives. OA derivative (1 mmol) was
dissolved in MeOH (50 mL). To this soln., 10% Pd on carbon (30 mg) was added and the reaction was
subjected to 1 atm of H2. After stirring for 12 h at rt, the reaction mixture was filtered, and the filtrate
was concentrated to give the target product.
General procedure D, hydrolysis of OA derivatives. OA derivative (1 mmol) was dissolved in THF
(20 mL) and MeOH (20 mL). To this soln., 2 M NaOH solution (12 mL) was added. After stirring for 12
h at rt, the reaction mixture was poured into H2O (40 ml), the pH was adjusted to 6-7 by adding 5% HCl,
and the mixture was extracted with AcOEt (3×30 mL). The combined organic layer was washed with
brine, dried over anhydrous Na2SO4 and concentrated under reduced pressure. The residue was purified
by CC.
General procedure E, deprotection of TBS group of OA derivatives. OA derivative (1 mmol) was
o
dissolved in DCM (20 mL). To this soln., boron trifluoride ether etherate (6 mL) was added at 0 C.
o
After stirring for 5 h at 0 C, the reaction mixture was poured into H2O (40 mL), and the mixture was
extracted with DCM (3×20 mL). The combined organic layer was washed with brine, dried over
anhydrous Na2SO4 and concentrated in vacuo. The residue was purified by CC.
Compound 1. OA (10 g, 21.9 mmol) was dissolved in dry DMF (100 mL). To this soln., K2CO3 (6.04 g,
43.8 mmol) and BnBr (3.33 g, 26.3 mmol) were added. After stirring for 12 h at rt, the reaction mixture
was poured into ice H2O (200 mL). The precipitate was filtered and washed with water, then dried under
reduced pressure to give 1 as a white solid powder (11 g, 92%).
Compound 2. General procedure A, CC (PE/AcOEt 10:1), afforded 2 (840 mg, 85%) as a white solid.
1H NMR (400 MHz, CDCl3): 7.34-7.35 (m, 5 H); 5.31 (s, 1 H); 5.04-5.12 (m, 2 H); 2.90-2.93 (m, 1 H);
2.50-2.54 (m, 1 H); 2.34-2.37 (m, 1 H); 1.18-1.99 (m, 20 H); 1.13 (s, 3 H); 1.08 (s, 3 H); 1.03 (s, 3 H);
1.01 (s, 3 H); 0.92 (s, 3 H); 0.90 (s, 3 H); 0.65 (s, 3 H).
Compound 3. t-BuOK (515 mg, 4.2 mmol) and 2 (500 mg, 0.92 mmol) were dissolved in t-BuOH (30

HETEROCYCLES, Vol. 85, No. 5, 2012
1127
mL) and stirred for 1 h at rt. To this soln. isoamyl nitrite (493 mg, 4.2 mmol) was added. After stirring
for 5 h at rt, the reaction mixture was concentrated under reduced pressure. The residue was purified by
1
CC (PE/AcOEt 5:1) to give 3 (450 mg, 86%) as a white solid. H NMR (400 MHz, CDCl3): 7.29-7.32
(m, 5 H); 5.31 (s, 1 H); 5.00-5.09 (m, 2 H); 2.96-3.00 (m, 1 H); 2.89-2.92 (m, 1 H); 1.19-2.11 (m, 19 H);
1.16 (s, 3 H); 1.12 (s, 3 H); 1.09 (s, 3 H); 0.90 (s, 3 H); 0.88 (s, 3 H); 0.85 (s, 3 H); 0.61 (s, 3 H).
Compound 4. Hydroxylamine hydrochloride (165 mg, 2.35 mmol) and 3 (450 mg, 0.78 mmol) were
dissolved in pyridine (20 mL). After refluxing for 4 h, the reaction mixture was concentrated under
reduced pressure. The residue was purified by CC (PE/AcOEt 3:1) to give 4 (345 mg, 75%) as a white
solid. 1H NMR (400 MHz, CDCl3): 7.32-7.35 (m, 5 H); 5.33 (s, 1 H); 5.04-5.12 (m, 2 H); 3.10-3.15 (m,
1 H); 2.90-2.93 (m, 1 H); 1.21-2.05 (m, 19 H); 1.18 (s, 3 H); 1.12 (s, 3 H); 1.07 (s, 3 H); 0.90 (s, 3 H);
0.89 (s, 3 H); 0.85 (s, 3 H); 0.63 (s, 3 H).
Compound 5. KOH (32 mg, 0.58 mmol) and 4 (345 mg, 0.58 mmol) were dissolved in ethylene glycol
(10 mL) and dioxane (5 mL). After refluxing for 5 h, the reaction mixture was poured into H2O (30 mL)
and extracted with AcOEt (3×30 mL). The combined organic layer was washed with brine, dried over
anhydrous Na2SO4 and concentrated under reduced pressure. The residue was purified by CC
(PE/AcOEt 10:1) to give 5 (200 mg, 60%) as a white solid. 1H NMR (400 MHz, CDCl3): 7.32-7.36 (m,
5 H); 5.36 (s, 1 H); 5.04-5.12 (m, 2 H); 3.06-3.10 (m, 1 H); 2.94-2.96 (m, 1 H); 2.13-2.17 (m, 1 H);
1.46-1.99 (m, 18 H); 1.41 (s, 3 H); 1.33 (s, 3 H); 1.12 (s, 3 H); 0.93 (s, 3 H); 0.90 (s, 3 H); 0.81 (s, 3 H);
0.65 (s, 3 H).
1
Compound 6. General procedure C, afforded 6 (168 mg, 92%) as a white solid. Mp 243-245 oC. H
NMR (400 MHz, CDCl3): 5.35 (s, 1 H); 3.08-3.12 (m, 1 H); 2.86-2.89 (m, 1 H); 2.15-2.19 (m, 1 H);
1.44-2.02 (m, 18 H); 1.41 (s, 3 H); 1.30 (s, 3 H); 1.17 (s, 3 H); 0.93 (s, 3 H); 0.91 (s, 3 H); 0.84 (s, 3 H);
13
0.82 (s, 3 H). C NMR (CDCl3, 100 MHz): 184.3; 159.6; 150.5; 143.6; 122.0; 52.9; 46.6; 45.9; 45.8;
41.8; 41.0; 39.3; 38.4; 35.2; 33.8; 33.2; 33.0; 32.4; 31.7; 31.3; 30.7; 29.7; 27.7; 25.7; 24.7; 23.5; 22.9;
19.0; 16.6; 15.4. ESI-HRMS (m/z) [M+Na]+ calcd for C30H44N2NaO3 503.3244; found 503.3284.
Compound 7. Zn dust (70 mg, 1.1 mmol) and 3 (200 mg, 0.35 mmol) were dissolved in AcOH (10 mL)
and Ac2O (5 mL). After stirring for 12 h at rt, the reaction mixture was filtered with diatomite, the pH of
filtrate was adjusted to 6-7 by adding 5% HCl, and the mixture was extracted with AcOEt (3×30 mL).
The combined organic layer was washed with brine, dried over anhydrous Na2SO4 and concentrated
under reduced pressure. The residue was purified by CC (PE/AcOEt 1:1) to give 7 (170 mg, 86%) as a
1
white solid. H NMR (400 MHz, CDCl3): 7.27-7.32 (m, 5 H); 6.58 (d, J = 6.0 Hz, 1 H); 5.26 (s, 1 H);
5.05 ( d, J = 12.0 Hz, 1 H); 5.00 ( d, J = 12.0 Hz, 1H); 4.84-4.88 (m, 2H); 2.87-2.90 (m, 1H); 2.54-2.61
(m, 1H); 2.04 (s, 3H); 1.19-2.02 (m, 19 H); 1.14 (s, 3 H); 1.11 (s, 3 H); 1.07 (s, 3 H); 0.90 (s, 3 H); 0.89
(s, 3 H); 0.74 (s, 3 H); 0.57 (s, 3 H).

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Compound 8. POCl3 (0.5 mL) and 7 (170 mg, 0.28 mmol) were dissolved in pyridine (10 mL). After
stirring for 12 h at rt, H2O (20 mL) was added in to quench the reaction and the mixture was extracted
with AcOEt (3×30 mL). The combined organic layer was washed with brine, dried over anhydrous
Na2SO4 and concentrated under reduced pressure. The residue was purified by CC (PE/AcOEt 5:1) to
1
give 8 (125 mg, 75%) as a white solid. H NMR (400 MHz, CDCl3): 7.22-7.27 (m, 5 H); 5.27 (s, 1 H);
5.01 (d, J = 12 Hz, 1 H); 4.96 (d, J = 12 Hz, 1 H); 2.85-2.88 (m, 1 H); 2.32-2.36 (m, 1 H); 2.31 (s, 3 H);
1.19-2.02 (m, 19 H); 1.12 (s, 3 H); 1.10 (s, 3 H); 1.03 (s, 3 H); 0.85 (s, 3 H); 0.83 (s, 3 H); 0.82 (s, 3 H);
0.59 (s, 3 H).
Compound 9. General procedure C, afforded 9 (83 mg, 82%) as a white solid. Mp 267-268 oC. 1H NMR
(500MHz, CDCl3): 5.32 (s, 1 H); 2.84-2.86 (m, 1 H); 2.39 (s, 3 H); 1.24-2.08 (m, 20 H); 1.17 (s, 3 H);
1.13 (s, 3 H); 1.07 (s, 3 H); 0.91 (s, 3 H); 0.88 (s, 3 H); 0.81 (s, 3 H); 0.82 (s, 3 H). 13C NMR (100 MHz,
CDCl3): 183.4; 159.8; 151.9; 143.6; 129.8; 122.4; 53.5; 53.4; 46.5; 46.2; 45.9; 41.7; 41.0; 39.4; 39.0;
38.5; 33.8; 33.0; 32.4; 31.9; 30.6; 28.8; 27.7; 25.7; 23.5; 23.3; 22.9; 21.0; 18.8; 16.8; 15.7; 14.0.
ESI-HRMS (m/z) [M+H]+ calcd for C32H48NO3 494.3629; found 494.3695.
Compound 10. General procedure A, afforded 10 (1.0 g, 90%) as a white solid and used without further
purification.
Compound 11. MeONa (595 mg, 11 mmol) and 10 (1.0 g, 2.2 mmol) were dissolved in dry toluene (30
mL). After stirring for 1 h at rt, HCO2Et (0.89 mL, 11 mmol) was added. After stirring for 6 h, the
reaction mixture was concentrated under reduced pressure. The residue was diluted with H2O (30 mL)
and adjusted pH to 6-7 by adding 5% HCl, and the mixture was extracted with AcOEt (3×30 mL). The
combined organic layer was washed with brine, dried over anhydrous Na2SO4 and concentrated under
reduced pressure. The residue was purified by CC (PE/AcOEt 10:1) to give 11 (743 mg, 70%) as a pink
solid. 1H NMR (500 MHz, CDCl3): 14.92 (d, J = 4.0 Hz, 1 H), 8.57 (d, J = 4.0 Hz, 1 H), 5.34 (m, 1 H),
2.90 (m, 1 H), 1.20-2.87 (m, 21 H); 1.19 (s, 3 H), 1.14 (s, 3 H), 1.10 (s, 3 H), 0.94 (s, 3 H), 0.90 (s, 3 H),
0.89 (s, 3 H), 0.79 (s, 3 H).
Compound 12. General procedure B, CC (PE/AcOEt 5:1), afforded 2 (430 mg, 78%) as a pink solid. 1H
NMR (500 MHz, CDCl3): 15.71 (s, 1H, CO2H); 5.33 (s, 1H, H-12); 2.83-2.86 (m, 1H, H-18); 2.50-2.53
(m, 1H); 0.82-2.04 (m, 20H); 1.25 (s, 3 H); 1.17 (s, 3 H); 1.10 (s, 3 H); 0.93 (s, 3 H); 0.90 (s, 3 H); 0.89
(s, 3 H); 0.81 (s, 3 H).
Compound 13. General procedure B, CC (PE/AcOEt 5:1), afforded 13 (410 mg, 78%) as a white solid.
1H NMR (500 MHz, CDCl3): 12.57 (s, 1H, CO2H); 5.32 (s, 1H, H-12); 4.14-4.27 (m, 2H, CH2O); 2.
82-2.85 (m, 1H, H-18); 2.32-2.35 (m, 1H); 0.85-2.03 (m, 23H); 1.16 (s, 3H); 1.13 (s, 3H); 1.07 (s, 3H);
0.92 (s, 6H); 0.90 (s, 3H); 0.85 (s, 3H).
Compound 14. General procedure B, CC (PE/AcOEt 5:1), afforded 14 (410 mg, 78%) as a white solid.

HETEROCYCLES, Vol. 85, No. 5, 2012
1129
1H NMR (400 MHz, CDCl3): 5.24 (s, 1 H); 4.25-4.32 (m, 2 H); 2.86-2.89 (m, 1 H); 2.32-2.36 (m, 1 H);
1.27-1.91 (m, 26 H); 1.24 (s, 3 H); 1.14 (s, 3 H); 1.08 (s, 3 H); 0.83 (s, 3 H); 0.77 (s, 3 H); 0.60 (s, 3 H).
Compound 15. Hydroxylamine hydrochloride (210 mg, 3.0 mmol) and 11 (500 mg, 1.0 mmol) were
dissolved in pyridine (20 mL). After refluxing for 5 h, the reaction mixture was concentrated under
reduced pressure. The residue was purified by CC (PE/AcOEt 4:1) to give 15 (370 mg, 74%) as a white
1
solid. Mp 159-162 oC. H NMR (400 MHz, CDCl3): 8.03 (s, 1 H); 5.33 (s, 1 H); 2.84-2.86 (m, 1 H);
2.65-2.69 (m, 1 H); 1.25-2.04 (m, 19 H); 1.36 (s, 3 H); 1.23 (s, 3 H); 1.15 (s, 3 H); 0.93 (s, 3 H); 0.90 (s,
3 H); 0.81 (s, 3 H); 0.78 (s, 3 H). 13C NMR (100 MHz, CDCl3): 184.1; 167.7; 153.5; 143.6; 122.3;
113.0; 53.2; 46.6; 45.9; 45.8; 41.8; 41.0; 39.2; 38.1; 34.0; 33.8; 33.6; 33.0; 32.3; 31.9; 31.4; 30.6; 27.7;
25.7; 24.7; 23.5; 23.3; 22.9; 19.0; 16.7; 14.9. ESI-HRMS (m/z) [M+H]+ calcd for C31H46NO3 480.3478;
found 480.3448.
Compound 16. 85% hydrazine hydrate (270 mg, 4.5 mmol) and 12 (500 mg, 0.91 mmol) were dissolved
in dry EtOH (20 mL). After refluxing for 5 h, the reaction mixture was concentrated under reduced
pressure. The residue was purified by CC (DCM/MeOH 40:1) to give 16 (400 mg, 81%) as a white solid.
Mp 254-256 oC. 1H NMR (400 MHz, DMSO-d6): 13.2 (bs, 1 H); 12.1 (bs, 1 H); 5.21 (s, 1 H); 2.75-2.77
(m, 1 H); 2.49-2.52 (m, 1 H); 1.22-2.05 (m, 22 H); 1.13 (s, 3 H); 1.10 (s, 3 H); 1.05 (s, 3 H); 0.86 (s, 6
13
H); 0.80 (s, 3 H); 0.77 (s, 3 H). C NMR (100 MHz, DMSO-d6): 178.5; 147.8; 143.6; 123.8; 121.4;
121.2; 110.2; 52.1; 45.6; 45.5; 45.4; 41.5; 40.9; 40.1; 37.8; 34.9; 33.3; 32.8; 32.6; 32.0; 31.6; 30.3; 30.2;
27.2; 25.3; 23.2; 23.0; 22.8; 22.6; 18.4; 16.5; 15.1. ESI-HRMS (m/z) [M+Na]+ calcd for
C32H45F3N2NaO2 569.3325; found 569.3344.
Compound 17. 85% hydrazine hydrate (140 mg, 2.4 mmol) and 12 (300 mg, 0.56 mmol) were dissolved
in dry EtOH (20 mL). After refluxing for 5 h, the reaction mixture was concentrated under reduced
pressure. The residue was purified by CC (DCM/MeOH 20:1) to give 17 (194 mg, 67%) as a white solid.
Mp 265-266 oC. 1H NMR (400 MHz, DMSO-d6): 5.22 (s, 1 H); 2.75-2.78 (m, 1 H); 2.25-2.28 (m, 1 H);
1.31-1.97 (m, 24 H); 1.15 (s, 3 H); 1.10 (s, 3 H); 1.05 (s, 3 H); 0.86 (s, 6 H); 0.79 (s, 3 H); 0.76 (s, 3 H).
13C NMR (100 MHz, DMSO-d6): 178.7; 158.6; 146.8; 143.6; 121.7; 95.5; 52.7; 45.7; 45.5; 41.5; 40.9;
40.1; 38.8; 38.1; 34.6; 33.4; 33.0; 32.9; 32.1; 31.8; 30.4; 30.3; 27.3; 25.4; 23.3; 22.9; 22.7; 18.6; 16.6;
15.1; 14.1. ESI-HRMS (m/z) [M+H]+ calcd for C31H47N2O3 495.3581; found 495.3587.
Compound 18. Hydroxylamine hydrochloride (78 mg, 1.14 mmol), AcONa (93 mg, 1.14 mmol) and 13
(120 mg, 0.23 mmol) were dissolved in dry EtOH (15 mL). After refluxing for 24 h, the reaction mixture
was diluted with H2O (30 mL) and extracted with AcOEt (3×30 mL). The combined organic layer was
washed with brine, dried over anhydrous Na2SO4 and concentrated under reduced pressure. The residue
1
was purified by CC (DCM/MeOH 30:1) to give 18 (76 mg, 68%) as a white solid. Mp 217-218 oC. H
NMR (400 MHz, DMSO-d6): 5.27 (s, 1 H); 2.81-2.84 (m, 1 H); 2.10-2.14 (m, 1 H); 1.23-2.03 (m, 23 H);

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1.26 (s, 3 H); 1.16 (s, 3 H); 1.13 (s, 3 H); 0.93 (s, 6 H); 0.88 (s, 3 H); 0.82 (s, 3H). 13C NMR (100 MHz,
DMSO-d6): 178.6; 171.8; 169.7; 143.6; 121.5; 63.3; 52.1; 48.6; 45.7; 45.5(×2); 41.5; 40.9; 37.5; 34.1;
33.6; 33.4; 32.8; 32.1; 31.6; 30.4; 28.9; 27.3; 25.4; 23.3; 22.7; 21.6; 18.3; 16.5; 15.0; 14.1. ESI-HRMS
(m/z) [M+Na]+ calcd for C31H45NNaO4 518.3241; found 518.3263.
Compound 19. 85% hydrazine hydrate (100 mg, 1.71 mmol), and 14 (190 mg, 0.34 mmol) were
dissolved in AcOH (15 mL). After stirring for 12 h at rt, the reaction mixture was diluted with H2O (30
mL) and adjusted pH to 6-7 with NaHCO3 and extracted with AcOEt (3×30 mL). The combined organic
layer was washed with brine, dried over anhydrous Na2SO4 and concentrated under reduced pressure.
The residue was purified by CC (DCM/MeOH 20:1) to give 19 (150 mg, 78%) as a white solid. Mp
1
311-312 oC. H NMR (400 MHz, DMSO-d6): 5.23 (s, 1 H); 4.19-4.21 (m, 2 H); 2.78-2.85 (m, 2 H);
1.25-2.03 (m, 25 H); 1.23 (s, 3 H); 1.20 (s, 3 H); 1.11 (s, 3 H); 1.10 (s, 6 H); 0.85 (s, 3 H); 0.77 (s, 3 H).
13C NMR (100 MHz, DMSO-d6): 178.6; 169.2; 150.5; 143.7; 138.9; 121.5; 119.2; 59.6; 52.4; 45.7; 45.5;
41.5; 40.9; 40.1; 38.8; 37.7; 36.6; 33.4; 32.8; 32.1; 31.7; 30.6; 30.4; 27.3; 27.2; 25.4; 23.3; 23.2; 22.9;
22.7; 18.6; 16.5; 15.3; 14.3. ESI-HRMS (m/z) [M+Na]+ calcd for C34H50N2NaO4 573.3663; found
573.3663.
Compound 20. General procedure D, CC (DCM/MeOH 10:1), afforded 20 (277 mg, 53%) as a white
1
solid. Mp 274-276 oC. H NMR (500 MHz, DMSO-d6): 12.45 (bs, 3 H); 5.24 (s, 1H); 2.92 (d, J = 16.0
Hz, 1 H); 2.78 (d, J = 16.0 Hz, 1 H); 1.33-2.08 (m, 19 H); 1.23 (s, 3 H); 1.14 (s, 6 H); 0.89 (s, 6 H); 0.80
13
(s, 6 H). C NMR (125 MHz, DMSO-d6): 178.6; 163.3; 150.5; 143.6; 138.9; 121.6; 115.3; 52.4; 45.7;
45.5; 41.5; 40.9; 40.0; 39.0; 38.9; 37.7; 36.6; 33.4; 32.9; 32.8; 32.1; 31.8; 30.8; 30.4; 27.2; 25.4; 23.4;
23.3; 23.0; 22.7; 16.6; 15.3. ESI-HRMS (m/z) [M+Na]+ calcd for C32H46N2NaO4 545.3350; found
545.3355.
Compound 21. Hydroxylamine hydrochloride (123 mg, 1.8 mmol) and 14 (200 mg, 0.36 mmol) were
dissolved in dry EtOH (15 mL). After refluxing for 5 h, the reaction mixture was diluted with H2O (30
mL) and extracted with AcOEt (3×20 mL). The combined organic layer was washed with brine, dried
over anhydrous Na2SO4 and concentrated under reduced pressure. The residue was purified by CC
1
(DCM/MeOH 40:1) to give 21 (150 mg, 75%) as a white solid. Mp 170-172 oC. H NMR (500 MHz,
DMSO-d6): 5.32 (s, 1 H); 4.37-4.41 (m, 2 H); 2.82-2.85 (m, 1 H); 2.77-2.76 (m, 1 H); 1.37-2.09 (m, 22
H); 1.32 (s, 3 H); 1.17 (s, 3 H); 1.13 (s, 3 H); 0.91 (s, 3 H); 0.89 (s, 3 H); 0.86 (s, 3 H); 0.79 (s, 3 H). 13C
NMR (100 MHz, DMSO-d6): 183.2; 175.0; 159.7; 153.0; 142.3; 121.2; 109.7; 60.5; 59.2; 52.2; 51.8;
45.4; 45.0; 44.6; 40.6; 39.8; 38.2; 37.3; 34.5; 33.8; 32.7; 31.9; 31.3; 30.7; 29.5; 27.6; 26.5; 24.6; 21.7;
19.8; 17.5; 15.6; 14.3; 13.0. ESI-HRMS (m/z) [M+Na]+ calcd for C34H49NNaO5 574.3503; found
574.3550.
Compound 22. General procedure D, CC (DCM/MeOH 10:1), afforded 22 (288 mg, 55%) as a white

HETEROCYCLES, Vol. 85, No. 5, 2012
1131
solid. Mp 163-164 oC. 1H NMR (500 MHz, DMSO-d6): 5.24 (s, 1 H); 2.78-2.80 (m, 1 H); 2.65-2.69 (m,
2H); 1.28-2.09 (m, 18 H); 1.24 (s, 3 H); 1.09 (s, 3 H); 1.07 (s, 3 H); 0.89 (s, 6 H); 0.83 (s, 3 H); 0.80 (s,
3 H). 13C NMR (100 MHz, DMSO-d6): 178.5; 175.2; 161.6; 154.8; 143.7; 121.3; 110.3; 59.7; 52.1; 45.7;
45.5; 45.4; 41.5; 37.9; 35.1; 34.4; 33.3; 32.8; 32.0; 31.4; 30.3; 28.4; 27.2; 25.3; 23.3; 22.9; 22.6; 20.7;
18.1; 16.5; 15.1; 14.0. ESI-HRMS (m/z) [M+Na]+ calcd for C32H45NNaO5 546.3190; found 546.3182.
Compound 23. DMAP (0.45 g, 3.66 mmol), 1 (10 g, 18.3 mmol), Ac2O (3.73g, 36.6 mmol) and
pyridine (4.4 mL, 55 mmol) were dissolved in dry DMF (50 mL). After stirring at rt for 12 h, the
reaction mixture was poured into ice water (100 mL) and the mixture was extracted with AcOEt (3×30
mL). The combined organic layer was washed with aqueous NaHCO3, 5% HCl and brine, dried over
anhydrous Na2SO4 and concentrated under reduced pressure afforded 23 (9.7 g, 90%) as a white solid
and used without further purification.
Compound 24. Chromium anhydride (1.02 g, 10.2 mmol) was dissolved in DCM (20 mL). To the soln.
pyridine (1.64 mL, 20.4 mmol) was slowly added at 0 oC and the reaction mixture was stirred for 0.5 h,
then 23 (300 mg, 0.51 mmol) was added. After stirring for 12 h at rt, the solution was filtered through
silica gel, the filtrate was concentrated under reduced pressure and dissolved in AcOEt (100 mL),
washed with saturated aqueous NaHSO3 and brine, dried over anhydrous Na2SO4 and concentrated under
reduced pressure. The residue was purified by CC (PE/AcOEt 5:1) to give 24 (153mg, 50%) as a white
solid. 1H NMR (400 MHz, CDCl3): 7.30-7.32 (m, 5 H); 5.61 (s, 1 H); 5.03-5.12 (m, 2 H); 4.46 (dd, J =
4.0, 11.5 Hz, 1 H); 3.05-3.07 (m, 1 H); 2.77-2.79 (m, 1 H); 2.05 (s, 3 H); 1.17-1.94 (m, 20 H); 1.16 (s, 3
H); 1.0 (s, 3 H); 0.94 (s, 3 H); 0.91 (s, 3 H); 0.89 (s, 3 H); 0.88 (s, 3 H); 0.86 (s, 3 H).
Compound 25. General procedure D and C, afforded 25 (377 mg, 80%, over two steps) as a white solid.
Mp 279-280 oC. 1H NMR (500 MHz, CDCl3): 12.33 (bs, 1 H); 5.43 (s, 1 H); 4.24 (s, 1 H); 3.17 (m, 1 H);
3.0-3.02 (m, 1 H); 2.84-2.86 (m, 1 H); 2.59-2.62 (m, 1 H); 2.29 (s, 1 H); 1.34-2.08 (m, 17 H); 1.33 (s, 3
H); 1.01 (s, 3 H); 0.90 (s, 6 H); 0.88 (s, 3 H); 0.87 (s, 3 H); 0.66 (s, 3 H). 13C NMR (125 MHz,
DMSO-d6): 199.2; 178.1; 169.1; 126.8; 76.6; 61.1; 54.2; 45.1; 44.6; 43.8; 43.2; 41.2; 38.7; 38.5; 36.9;
36.8; 33.1; 32.5; 31.2; 30.3; 28.1; 27.3; 26.9; 23.2; 23.1; 22.4; 18.8; 17.1; 16.0; 15.9. ESI-HRMS (m/z)
[M+Na]+ calcd for C30H46NaO4 493.3288; found 493.3268.
Compound 26. mCPBA (1 g, 5.1 mmol) and 23 (1 g, 1.7 mmol) were dissolved in CHCl3 (20 mL).
After stirring for 24 h at rt, the reaction mixture was diluted with H2O (30 mL) and extracted with CHCl3
(2×30 mL). The combined organic layer was washed with saturated aqueous Na2SO3 and brine, dried
over anhydrous Na2SO4 and concentrated under reduced pressure. The residue was purified by CC
(PE/AcOEt 10:1) to give 26 (616 mg, 60%) as a white solid. 1H NMR (400 MHz, CDCl3): 7.31-7.34 (m,
5 H); 5.19 (d, J = 12.2 Hz, 1 H); 5.06 (d, J = 12.2 Hz, 1 H); 4.45 (dd, J = 5.1, 12.0 Hz, 1 H); 2.80-2.84
(m, 1 H); 2.43 (d, J = 5.2 Hz, 1 H); 2.18-2.19 (m, 1 H); 2.15 (s, 3 H); 1.15-2.08 (m, 21 H); 1.12 (s, 3 H);

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0.98 (s, 3 H); 0.92 (s, 3 H); 0.89 (s, 3 H); 0.88 (s, 3 H); 0.77 (s, 3 H); 0.61 (s, 3 H).
Compound 27. General procedure D and C, afforded 27 (364 mg, 77%, over two steps) as a white solid.
1
Mp 223-224 oC. H NMR (500 MHz, CDCl3): 3.16-3.20 (m, 1 H); 2.67-2.69 (m, 1 H); 2.42-2.45 (m, 1
H); 2.19 (s, 1H); 1.14-2.15 (m, 20 H); 1.12 (s, 3 H); 0.98 (s, 3 H); 0.92 (s, 3 H); 0.89 (s, 3 H); 0.88 (s, 3
13
H); 0.77 (s, 3 H); 0.61 (s, 3 H). C NMR (100 MHz, DMSO-d6): 209.3; 177.5; 75.0; 52.9; 49.4; 47.6;
44.4; 39.8; 39.2; 36.8; 36.4; 35.9; 34.8; 34.2; 32.4; 31.7; 31.0; 30.0; 29.8; 29.0; 28.7; 26.4; 25.5; 25.3;
21.5; 20.8; 18.5; 16.4; 14.2; 13.3. ESI-HRMS (m/z) [M+Na]+ calcd for C30H48NaO4 495.3445; found
495.3457.
Compound 28. 2 M solution of bromine in AcOH (0.5 mL), 26 (300 mg, 0.49 mmoL) and several drops
of hydrobromic acid were added to glycial AcOH (15 mL). The reaction mixture was heated for 4 h at
60 oC. The reaction mixture was quenched with H2O (30 mL) and adjusted pH to 7-8 by adding saturated
aqueous NaHCO3 solution, then extracted with AcOEt (3 ×30 mL). The combined organic layer was
washed with brine, dried over anhydrous Na2SO4 and concentrated under reduced pressure. The residue
1
was purified by CC (PE/AcOEt 10:1) to give 28 (230 mg, 78%) as a white solid. H NMR (500 MHz,
CDCl3): 7.28-7.36 (m, 5 H); 5.71 (s, 1 H); 5.14 (d, J = 12.0 Hz, 1 H); 5.10 (d, J = 12.0 Hz, 1 H); 4.46
(dd, J = 5.0, 12.0 Hz, 1 H); 3.05-3.08 (m, 1 H); 2.73 (d, J = 4.4 Hz, 1 H); 2.06 (s, 3 H); 0.79-1.94 (m, 19
H); 1.16 (s, 3 H); 1.0 (s, 3 H); 0.94 (s, 3 H); 0.91 (s, 3 H); 0.89 (s, 3 H); 0.88 (s, 3 H); 0.86 (s, 3 H).
Compound 29. General procedures D and C, afforded 29 (348 mg, 74%, over two steps) as a white solid.
Mp 299-301 oC. 1H NMR (500 MHz, CDCl3): 5.75 (s, 1 H); 3.22 (dd, J = 5.0, 11.7 Hz, 1 H); 2.98-3.01
(m, 1 H); 2.89-2.90 (m, 1 H); 1.18-1.95 (m, 19 H); 1.17 (s, 3 H); 1.03 (s, 3 H); 1.00 (s, 3 H); 0.94 (s, 3
13
H); 0.89 (s, 3 H); 0.87 (s, 3 H); 0.83 (s, 3 H). C NMR (100 MHz, DMSO-d6): 197.6; 177.2; 176.9;
120.1; 74.3; 48.1; 47.0; 44.3; 43.2; 39.5; 38.5; 37.3; 34.4; 33.7; 32.3; 31.5; 30.8; 30.7; 29.4; 28.6; 26.4;
26.0; 25.6; 22.0; 21.9; 21.3; 20.6; 19.5; 15.9; 14.3. ESI-HRMS (m/z) [M+Na]+ calcd for C30H46NaO4
493.3194; found 493.3174.
Compound 30. Hydroxylamine hydrochloride (58 mg, 0.83 mmol) and 26 (100 mg, 0.17 mmol) were
dissolved in dry pyridine (20 mL). After refluxing for 5 h, the reaction mixture was concentrated under
reduced pressure and diluted with H2O (30 mL), then extracted with AcOEt (3×20 mL). The combined
organic layer was washed with brine, dried over anhydrous Na2SO4 and concentrated under reduced
pressure afforded 30 (100 mg, 97%) as a white solid and used without further purification.
Compound 31. General procedure D and C, afforded 31 (370 mg, 76%, over two steps) as a white solid.
1
Mp 184-185 oC. H NMR (400 MHz, CDCl3): 3.18 (dd, J = 5.0, 15.0 Hz, 1 H); 3.0-3.03 (m, 1 H);
2.79-2.82 (m, 1 H); 2.53-2.55 (m, 1 H); 1.23-1.97 (m, 21 H); 1.21 (s, 3 H); 0.98 (s, 3 H); 0.93 (s, 3 H);
0.90 (s, 3 H); 0.88 (s, 3 H); 0.85 (s, 3 H); 0.73 (s, 3 H). 13C NMR (100 MHz, DMSO-d6): 179.5; 157.0;
76.7; 54.6; 47.2; 46.8; 42.5; 40.7; 40.1; 38.9; 38.4; 37.9; 36.6; 35.7; 34.2; 33.5; 33.0; 32.2; 31.6; 30.5;

HETEROCYCLES, Vol. 85, No. 5, 2012
1133
28.0; 27.0; 23.1; 22.4; 20.0; 19.3; 18.0; 15.8; 15.5; 15.0. ESI-HRMS (m/z) [M+Na]+ calcd for
C30H49NNaO4 510.3629; found 510.3633.
Compound 32. Compound 28 (250 mg, 0.4 mmol) was dissolved in dry THF (20 mL). To the soln.
SOCl2 (0.29 mL, 4.0 mmol) was added. After stirring for 12 h at rt, the reaction mixture was
concentrated under reduced pressure and diluted with H2O (30 mL), then extracted with AcOEt (3×20
mL). The combined organic layer was washed with brine, dried over anhydrous Na2SO4 and
concentrated under reduced pressure. The residue was purified by CC (DCM/MeOH 30:1) to give 32
(215 mg, 86%) as a white solid. 1H NMR (500 MHz, CDCl3): 7.33-7.37 (m, 5 H); 5.48 (d, J = 5.0 Hz, 1
H); 5.22 (d, J = 12.2 Hz, 1 H); 5.09 (d, J = 12.2 Hz, 1 H); 4.47 (dd, J = 5.0, 11.7 Hz, 1 H); 3.89-3.92 (m,
1 H); 2.52-2.55 (m, 1 H); 2.25-2.28 (m, 1 H); 2.03 (s, 3 H); 1.01-1.99 (m, 21 H); 0.94 (s, 3 H); 0.86 (s, 3
H); 0.83 (s, 3 H); 0.80 (s, 3 H); 0.77 (s, 3 H); 0.74 (s, 3 H); 0.70 (s, 3 H).
Compound 33. General procedure D and C, afforded 33 (332 mg, 68%, over two steps) as a white solid.
Mp 172-173 oC. 1H NMR (400 MHz, CDCl3): 6.12 (d, J = 5.0 Hz, 1 H); 4.06-4.09 (m, 1 H); 3.15 (dd, J
= 5.0, 11.5 Hz, 1 H); 2.52-2.54 (m, 1 H); 2.26-2.29 (m, 1 H); 0.95-1.93 (m, 20 H); 0.94 (s, 3 H); 0.86 (s,
3 H); 0.83 (s, 3 H); 0.80 (s, 3 H); 0.77 (s, 3 H); 0.74 (s, 3 H); 0.70 (s, 3 H). 13C NMR (100 MHz,
DMSO-d6): 176.9; 173.9; 74.9; 52.2; 48.5; 46.8; 44.5; 43.0; 40.7; 37.2; 36.9; 36.7; 36.6; 34.4; 32.9;
32.7; 31.7; 31.5; 31.3; 30.8; 28.9; 26.4; 25.2; 21.2; 19.7; 16.1; 15.6; 15.1; 14.2; 13.8. ESI-HRMS (m/z)
[M+Na]+ calcd for C30H49NNaO4 510.3554; found 510.3572.
Compound 34. Imidazole (1.3 g, 18.3 mmol), 1 (1.0 g, 1.83 mmol) and TBSCl (1.38 g, 9.16 mmol)
were dissolved in dry DMF (25 mL). After stirring for 6 h at 60 oC, the reaction mixture was poured into
H2O (30 mL), then extracted with AcOEt (3×30 mL). The combined organic layer was washed with
brine, dried over anhydrous Na2SO4 and concentrated under reduced pressure. The residue was
1
recrystallized with AcOEt to give 34 (1.0 g, 83%) as a white solid. H NMR (400 MHz, CDCl3):
7.29-7.30 (m, 5 H); 5.24 (s, 1H); 5.04 (d, J = 12.4 Hz, 1 H); 4.99 (d, J = 12.4 Hz, 1 H); 3.11-3.14 (m, 1
H); 2.84-2.87 (m, 1 H); 1.10-1.92 (m, 24 H); 1.07 (s, 3 H); 0.87 (s, 6 H); 0.85 (s, 3 H); 0.84 (s, 9 H);
0.82 (s, 3 H); 0.69 (s, 3 H); 0.50 (s, 3 H); -0.02 (s, 6H).
Compound 35. mCPBA (2.36 g, 6.82 mmol) and 34 (1.5 g, 2.27 mmol) were dissolved in CHCl3 (30
mL). After stirring for 12 h at rt, the reaction mixture was poured into saturated aqueous NaHSO3
solution (30 mL), and then extracted with AcOEt (3×30 mL). The combined organic layer was washed
with saturated aqueous Na2CO3 solution and brine, dried over anhydrous Na2SO4 and concentrated under
reduced pressure. The residue was purified by CC (PE/AcOEt 10:1) to give 35 (800 mg, 67%) as a white
1
solid. H NMR (400 MHz, CDCl3): 7.31-7.33 (m, 5 H); 5.05-5.21 (m, 2 H); 3.12-3.15 (m, 1 H);
2.77-2.80 (m, 1 H); 2.42-2.45 (m, 1 H); 2.17-2.19 (m, 1 H); 1.13-2.15 (m, 21 H); 1.12 (s, 3 H); 0.92 (s, 3
H); 0.90 (s, 3 H); 0.88 (s, 3 H); 0.87 (s, 12 H); 0.77 (s, 3 H); 0.61 (s, 3 H); 0.03 (s, 6 H).

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HETEROCYCLES, Vol. 85, No. 5, 2012
Compound 36 and 37. Compound 35 (900 mg, 1.33 mmol) was dissolved in THF (20 mL) and H2O (2
mL). To this soln. NaBH4 (150 mg, 4 mmol) was added. After stirring for 5 h, the pH of the reaction
mixture was adjusted to 6-7 by adding 5% HCl, and then the mixture was extracted with AcOEt (3×30
mL). The combined organic layer was washed with brine, dried over anhydrous Na2SO4 and
concentrated under reduced pressure. The residue was purified by CC (PE/AcOEt 15:1) to give 36 and
37 (804 mg, 89%, 36/37 = 2:3). 36: 1H NMR (500 MHz, CDCl3): 7.31-7.35 (m, 5 H); 5.17 (d, J = 12.3
Hz, 1 H); 5.07 (d, J = 12.3 Hz, 1 H); 3.99 (m, 1 H); 3.17 (dd, J = 4.1, 15.5 Hz, 1 H); 2.47-2.50 (m, 1 H);
1.29-2.38 (m, 23 H); 1.28 (s, 3 H); 0.93 (s, 3 H); 0.91 (s, 3 H); 0.88 (s, 12 H); 0.80 (s, 3 H); 0.72 (s, 3
H); 0.61 (s, 3H); 0.03 (s, 6 H). 13C NMR (100 MHz, CDCl3): 178.1; 136.4; 128.5(×2); 128.2; 128.1(×2);
79.3; 74.0; 66.0; 55.6; 48.5; 45.4; 42.0; 40.9; 40.4; 39.4; 39.2; 38.8; 36.6; 34.1; 33.5; 33.4; 33.2; 32.0;
30.7; 30.6; 28.4; 27.4; 27.3; 25.9; 23.7; 23.6; 22.7; 20.8; 18.4; 18.1; 16.5; 15.9; 15.8; 15.1; -3.8; -4.9. 37:
1H NMR (400 MHz, CDCl3): 7.31-7.40 (m, 5 H); 5.21 (d, J = 12.2 Hz, 1 H); 5.08 (d, J = 12.2 Hz, 1 H);
3.67 (m, 1 H); 3.14-3.16 (m, 1 H); 2.71-2.74 (m, 1 H); 1.01-2.69 (m, 23 H); 0.93 (s, 3 H); 0.92 (s, 3 H);
0.91 (s, 3 H); 0.88 (s, 12 H); 0.78 (s, 3 H); 0.72 (s, 3 H); 0.62 (s, 3H); 0.03 (s, 6 H). 13C NMR (100 MHz,
CDCl3): 177.9; 136.5; 128.5(×2); 128.4; 128.1(×2); 79.3; 68.3; 66.0; 55.3; 49.1; 47.4; 43.1; 41.5; 40.5;
39.4; 38.6; 36.8; 36.1; 34.5; 33.4; 33.1; 32.6; 31.9; 31.5; 30.5; 28.7; 28.4; 27.7; 25.9(×3); 23.4; 23.2;
18.4; 18.1; 17.8; 16.2; 15.9(×2); -3.8; -4.9.
1
Compound 39. General procedure E afforded 39 (474 mg, 84%) as a white solid. H NMR (500 MHz,
CDCl3): 7.30-7.39 (m, 5H); 5.18 (d, J = 12.0 Hz, 1 H); 5.06 (d, J = 12.0 Hz, 1 H); 3.67 (m, 1H); 3.17 (dd,
J = 4.8, 11.0 Hz, 1H); 2.72-2.78 (m, 1H); 1.13-2.03 (m, 23 H); 1.23 (s, 3 H); 0.95 (s, 3 H); 0.93 (s, 3 H);
0.91 (s, 3 H); 0.77 (s, 3 H); 0.74 (s, 3 H); 0.61 (s, 3 H).
Compound 40. General procedure E and C afforded 40 (370 mg, 78%, over two steps) as a white solid.
1
Mp 246-247 oC. H NMR (400 MHz, CDCl3): 3.77-3.80 (m, 1 H); 2.96-3.00 (m, 1 H); 2.24-2.27 (m, 1
H); 2.08-2.11 (m, 1 H); 1.07-1.97 (m, 27 H); 0.87 (s, 3 H); 0.86 (s, 3 H); 0.83 (s, 3 H); 0.80 (s, 3 H);
0.76 (s, 3 H); 0.64 (s, 3 H). 13C NMR (100 MHz, DMSO-d6): 179.8; 76.8; 71.6; 55.2; 47.3; 44.8; 41.6;
40.4; 38.5; 38.4; 38.3; 36.3; 33.9; 33.5; 33.0; 31.9; 30.6; 30.2; 30.0; 28.9; 28.7; 28.1; 27.0; 23.4; 20.1;
18.6; 18.0; 16.3; 16.0; 15.8. ESI-HRMS (m/z) [M+Na]+ calcd for C30H50NaO4 497.3601; found
497.3621.
Compound 41. General procedure E and C afforded 41 (356 mg, 75%, over two steps) as a white solid.
1
Mp 320-321 oC. H NMR (400 MHz, CDCl3): 3.73-3.76 (m, 1 H); 3.18-3.20 (m, 1 H); 2.70-2.76 (m, 1
H); 1.04-1.88 (m, 27 H); 0.93 (s, 6 H); 0.90 (s, 3 H); 0.83 (s, 3 H); 0.76 (s, 3 H); 0.68 (s, 3 H). 13C NMR
(100 MHz, DMSO-d6): 179.7; 77.1; 66.4; 55.2; 49.0; 46.5; 42.3; 41.4; 40.6; 38.9; 38.7; 36.9; 35.9; 34.6;
33.8; 33.4; 32.7; 32.0; 31.2; 30.6; 28.9; 28.5; 27.5; 23.6; 23.5; 18.4; 18.0; 16.5; 16.1; 16.0. ESI-HRMS
(m/z) [M+Na]+ calcd for C30H50NaO4 497.3601; found 497.3624.

HETEROCYCLES, Vol. 85, No. 5, 2012
1135
Compound 42. 60% NaH (35 mg, 0.9 mmol) and 37 (200 mg, 0.3 mmol) were dissolved in dry DMF
(15 mL). To this soln., MeI (0.06 mL, 0.9 mmol) was added. After stirring for 3 h at rt, the reaction
mixture was poured into H2O (30 mL), and then the mixture was extracted with AcOEt (3×30 mL). The
combined organic layer was washed with brine, dried over anhydrous Na2SO4 and concentrated under
reduced pressure. The residue was purified by CC (PE/AcOEt 10:1) to give 42 (174 mg, 85%). 1H NMR
(500 MHz, CDCl3): 7.29-7.40 (m, 5 H); 5.17 (d, J = 12.5 Hz, 1 H); 5.11 (d, J = 12.5 Hz, 1 H); 3.30 (s, 3
H); 3.21-3.24 (m, 1 H); 3.15 (dd, J = 4.4, 15.7 Hz, 1 H); 0.95-1.93 (m, 24 H); 0.92 (s, 3 H); 0.91 (s, 3
H); 0.90 (s, 3 H); 0.88 (s, 9 H); 0.87 (s, 3 H); 0.79 (s, 3 H); 0.72 (s, 3 H); 0.62 (s, 3 H); 0.03 (s, 6H).
Compound 43. General procedure E and C, afforded 43 (362 mg, 74%, over two steps) as a white solid.
Mp 274-276 oC. 1H NMR (500 MHz, CDCl3): 3.36 (s, 3 H); 3.26-3.30 (m, 1 H); 3.19 (dd, J = 4.6, 16.2
Hz, 1 H); 2.72-2.75 (m, 1 H); 0.98-2.03 (m, 23 H); 0.97 (s, 3 H); 0.96 (s, 3 H); 0.92 (s, 3 H); 0.91 (s, 3
H); 0.87 (s, 3 H); 0.83 (s, 3 H); 0.76 (s, 3 H). 13C NMR (100 MHz, DMSO-d6): 183.1; 92.0; 78.9; 55.9;
55.3; 48.9; 47.2; 41.5; 41.2; 40.4; 38.9; 38.7; 37.2; 36.5; 34.5; 33.5; 33.3; 32.5; 31.2; 30.6; 29.7; 28.8;
27.3; 26.0; 23.5; 23.1; 18.3; 17.9; 16.6; 15.9; 15.4. ESI-HRMS (m/z) [M+Na]+ calcd for C31H52NaO4
511.3758; found 511.3760.
Compound 44. Benzoyl chloride (0.35 mL, 3.0 mmol), 37 (200 mg, 0.3 mmol) and TEA (1.0 mL) were
dissolved in DCM (15 mL). After stirring for 5 h at rt, the reaction mixture was poured into H2O (30
mL), and then the mixture was extracted with DCM (3×20 mL). The combined organic layer was
washed with brine, dried over anhydrous Na2SO4 and concentrated under reduced pressure. The residue
1
was purified by CC (PE/AcOEt 10:1) to give 44 (180 mg, 78%) as a white solid. H NMR (400 MHz,
CDCl3): 7.96 (d, J = 7.3 Hz, 2 H); 7.46-7.48 (m, 1 H); 7.41 (d, J = 7.3 Hz, 2 H); 7.32-7.34 (m, 5 H); 5.24
(d, J = 12.2 Hz, 1 H); 5.11 (d, J = 12.2 Hz, 1 H); 3.16-3.18 (m, 1 H); 3.10-3.14 (m, 1 H); 2.70-2.72 (m, 1
H); 2.12-2.15 (m, 1 H); 1.13-2.11 (m, 22 H); 1.12 (s, 3 H); 0.92 (s, 3 H); 0.90 (s, 3 H); 0.87 (s, 12 H);
0.88 (s, 3 H); 0.77 (s, 3 H); 0.62 (s, 3 H); 0.03 (s, 6 H).
Compound 45. Chloroacetyl chloride (0.23 mL, 3.0 mmol), 37 (200 mg, 0.3 mmol) and pyridine (1.0
mL) were dissolved in DCM (15 mL). After stirring for 5 h at rt, the reaction mixture was poured into
H2O (30 mL), and then the mixture was extracted with DCM (3×20 mL). The combined organic layer
was washed with brine, dried over anhydrous Na2SO4 and concentrated under reduced pressure. The
residue was purified by CC (PE/AcOEt 10:1) to give 45 (200 mg, 90%) as a white solid. 1H NMR (400
MHz, CDCl3): 7.36-7.40 (m, 5 H); 5.20-5.29 (m, 2 H); 4.00 (s, 2 H); 3.10-3.13 (m, 1 H); 2.54-2.57 (m, 1
H); 1.15-2.50 (m, 24 H); 1.12 (s, 3 H); 0.97 (s, 3 H); 0.90 (s, 3 H); 0.87 (s, 12 H); 0.88 (s, 3 H); 0.77 (s,
3 H); 0.62 (s, 3 H); 0.03 (s, 6 H).
Compound 46. General procedure E and C, afforded 46 (432 mg, 75%, over two steps) as a white solid.
1
Mp 195-197 oC. H NMR (500 MHz, DMSO-d6): 12.25 (br, 1 H); 7.96-7.97 (m, 2 H); 7.60-7.63 (m, 1

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HETEROCYCLES, Vol. 85, No. 5, 2012
H); 7.48-7.51 (m, 2 H); 4.90-5.20 (m, 1 H); 2.90-3.00 (m, 1 H); 2.62-2.65 (m, 1 H); 2.10-2.40 (m, 1 H);
1.11-2.33 (m, 20 H); 1.01 (s, 3 H); 0.91 (s, 3 H); 0.88 (s, 3 H); 0.75 (s, 3 H); 0.72 (s, 3 H); 0.62 (s, 3 H);
0.45 (s, 3 H). 13C NMR (125 MHz, DMSO-d6): 183.3; 165.4; 133.1; 130.2; 129.0 (×2); 128.5 (×2); 76.6;
72.2; 54.7; 48.4; 46.1; 41.0; 40.3; 40.1; 39.9; 38.4; 38.2; 36.6; 36.1; 34.0; 33.2; 32.7; 32.1; 31.2; 30.1;
28.5; 28.1; 27.2; 27.0; 22.9; 17.9; 17.4; 16.2; 15.7; 15.6. ESI-HRMS (m/z) [M+Na]+ calcd for
C37H54NaO5 601.3863; found 601.3884.
Compound 47. General procedure E, afforded 47 (526 mg, 82%) as a white solid. 1H NMR (400 MHz,
CDCl3): 7.37-7.43 (m, 5 H); 5.08-5.28 (m, 2 H); 4.03 (s, 2 H); 3.17-3.20 (m, 1 H); 2.57-2.60 (m, 1 H);
1.25-2.43 (m, 24 H); 1.21 (s, 3 H); 0.99 (s, 3 H); 0.92 (s, 3 H); 0.85 (s, 3 H); 0.79 (s, 3 H); 0.77 (s, 3 H);
0.65 (s, 3 H).
Compound 48. Piperidine (0.27 mL, 3.1 mmol) and 47 (200 mg, 0.31 mmol) were dissolved in DCM
(20 mL). After refluxing for 6 h, the reaction mixture was poured into H2O (30 mL), and then the
mixture was extracted with DCM (3×20 mL). The combined organic layer was washed with brine, dried
over anhydrous Na2SO4 and concentrated under reduced pressure. The residue was purified by CC
(PE/AcOEt 5:1) to give 48 (182 mg, 85%) as a white solid. 1H NMR (500 MHz, CDCl3): 7.33-7.38 (m,
5 H); 5.04-5.23 (m, 3 H); 3.62-3.75 (m, 2 H); 3.22-3.50 (m, 4 H); 3.15-3.17 (m, 1 H); 2.43-2.45 (m, 1
H); 1.21-2.03 (m, 29 H); 0.94 (s, 6 H); 0.87 (s, 3 H); 0.77 (s, 3 H); 0.75 (s, 3 H); 0.73 (s, 3 H); 0.60 (s, 3
H).
Compound 49. Using the same procedure as preparation of 48 afforded 49 (181 mg, 85%) as a white
solid. 1H NMR (500 MHz, CDCl3): 7.33-7.38 (m, 5 H); 5.06-5.23 (m, 3 H); 4.04 (m, 4 H); 3.62-3.75 (m,
2 H); 3.22-3.50 (m, 4 H); 3.15-3.23 (m, 1 H); 2.39-2.42 (m, 1 H); 1.25-1.94 (m, 24 H); 1.24 (s, 3 H);
0.95 (s, 3 H); 0.89 (s, 3 H); 0.87 (s, 3 H); 0.79 (s, 3 H); 0.74 (s, 3 H); 0.64 (s, 3 H).
1
Compound 50. General procedure C, afforded 50 (516 mg, 86%) as a white solid. Mp 120-123 oC. H
NMR (500 MHz, DMSO-d6): 4.86-4.88 (m, 1 H); 3.17 (d, J = 17.0 Hz, 1 H); 3.00 (d, J = 17.0 Hz, 1 H);
2.96-2.99 (m, 1 H); 2.44-2.46 (m, 4 H); 2.02-2.06 (m, 1 H); 1.11-1.80 (m, 29 H); 0.96 (s, 3 H); 0.87 (s, 3
H); 0.85 (s, 6 H); 0.76 (s, 3 H); 0.75 (s, 3 H); 0.65 (s, 3 H). 13C NMR (100 MHz, DMSO-d6): 178.9;
169.7; 76.6; 71.1; 59.3; 54.8; 54.6; 53.2; 48.5; 48.2; 46.0; 41.0; 38.9; 38.4; 38.1; 36.5; 35.7; 33.8; 33.1;
32.7; 32.0; 30.9; 30.6; 30.1; 28.3; 28.0; 27.1; 27.0; 25.4; 23.5; 23.2; 22.6; 17.9; 17.3; 16.0; 15.7; 15.5.
ESI-HRMS (m/z) [M+H]+ calcd for C37H62NO5 600.4728; found 600.4705.
1
Compound 51. General procedure C, afforded 51 (512 mg, 85%) as a white solid. Mp 95-98 oC. H
NMR (500 MHz, DMSO-d6): 12.25 (br, 1 H); 4.87-4.90 (m, 1 H); 4.26-4.28 (m, 1 H); 3.56 (t, J = 4.5 Hz,
4 H); 3.25 (d, J = 16.9 Hz, 1 H); 3.07 (d, J = 16.9 Hz, 1 H); 2.96-2.99 (m, 1 H); 1.16-2.54 (m, 28 H);
0.96 (s, 3 H); 0.87 (s, 3 H); 0.85 (s, 3 H); 0.84 (s, 3 H); 0.76 (s, 3 H); 0.74 (s, 3 H); 0.64 (s, 3 H). 13C
NMR (125 MHz, DMSO-d6): 178.9; 169.5; 76.6; 71.2; 66.1; 58.7; 54.5; 52.5; 48.2; 46.0; 40.9; 40.2;

HETEROCYCLES, Vol. 85, No. 5, 2012
1137
39.2; 39.1; 39.0; 38.4; 38.1; 36.5; 35.7; 33.7; 33.1; 32.7; 32.0; 30.9; 30.1; 28.3; 28.0; 27.1; 27.0; 23.2;
22.6; 17.9; 17.3; 16.0; 15.7; 15.6. ESI-HRMS (m/z) [M+H]+ calcd for C36H60NO6 602.4415; found
602.4485.
In vitro assay of PTP1B inhibitors21
The tested compounds were solubilized in Me2SO at 5 mg/mL, and 2 μL samples was distributed to
A2-H11 wells of 96-well clear polystyrene plate (Corning, Action, MA). The Me2SO (2 μL) was
distributed to A1-D1 and E12-H12 wells as the full enzyme activity, and compound 52 was distributed
to E1-H1 and A12-D12 wells as the positive inhibition. After adding an assay mixture (80 μL), 10 μL of
the GST-PTP1B (300 nM) was added to initiate the reaction. The high-throughput screening was carried
out in a final 100 μL volume containing 50 mM MOPS, pH 6.5, 2 mM pNPP, 30 nM PTP1B and 2%
Me2SO, and the catalysis of pNPP was continuously monitored on SpectraMax 340 microplate reader at
405 nm for 2 min at 30 °C. For calculating IC50, inhibition assays were performed with 30 nM
recombinant enzyme, 2 mM pNPP in 50 mMMOPS at pH 6.5 and the inhibitors diluted around the
estimated IC50 values. IC50 was calculated from the non-linear curve fitting of percent inhibition (%
inhibition) vs. inhibitor concentration [I] by usingthe following equation % Inhibition=100/{1+( IC50/
[I])k}, where k is the Hill coefficient.
Crystal data and structure determination
A single crystal of 39 with dimensions of 0.398 mm × 0.369 mm × 0.307 mm was chosen for X-ray
diffraction analysis performed on a Bruker-AXS diffractometer, equipped with Mo Ka radiation ( =
0.71073 Å ) at 293(2) K by using a φ-ω scan mode. In the ranges of 1.73≤θ≤25.50°, a total of 35026
reflections were collected including 12334 unique ones (R int = 0.0793), of which 12334 were observed
with I > 2σ(I). The structure was solved by direct methods using SHELXS program of the SHELXL-97
package and refined with SHELXL. The final refinement was performed by full-matrix least-squares
method with Full-matrix least-squares on F2 for the non-hydrogen atoms. 39 (C37H56O4, Mr = 564.82),
crystallizes in the orthorhombic system, space group P212121 with a = 13.3536(9), b = 19.8860(12), c =
24.9647(16) Å,  = β = =90o, V = 6629.4(7) Å3, Z = 8, Dc = 1.132mg/m3, μ = 0.075mm−1, F(000) =
2480, the final R1 = 0.0705 and wR2 = 0.1778 for 12334 observed reflections (I > 2σ(I)). The hydrogen
atoms were located from Fourier difference maps. The final R1 = 0.0705, wR2 = 0.1778 (w = 1/[σ2(Fo2)
+ (0.1038P)2 + 0.0000P], where P = (Fo2 + 2Fc2)/3, S = 0.921, (Δρ) max = 0.592 and (Δρ) min =
-0.264 e/Å3.
ACKNOWLEDGEMENTS
This work was supported by the National Natural Science Foundation of China (Grant No. 20802020)

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and Ph.D. Program Foundation of Ministry of Education of China (Grant No. 200802691039).
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