An enantio-and diastereocontrolled synthesis of (-)-salinosporamide A

Article abstract of DOI:10.3987/COM-10-12042

The enantio-and diastereocontrolled total synthesis of (-)-salinosporamide A, a potent 20S proteasome inhibitor, was accomplished through organocatalytic aldolization, diastereoselective Claisen condensation, a Rh-catalyzed Reformatsky reaction, and an AZADO-catalyzed oxidative β-lactonization reaction as the key reactions. The Japan Institute of Heterocyclic Chemistry.

Full text of DOI:10.3987/COM-10-12042

HETEROCYCLES, Vol. 81, No. 10, 2010
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HETEROCYCLES, Vol. 81, No. 10, 2010, pp. 2239 - 2246. © The Japan Institute of Heterocyclic Chemistry
Received, 10th August, 2010, Accepted, 26th August, 2010, Published online, 26th August, 2010
DOI: 10.3987/COM-10-12042
AN ENANTIO- AND DIASTEREOCONTROLLED SYNTHESIS OF
(–)-SALINOSPORAMIDE A
Yosuke Sato, Hayato Fukuda, Masaki Tomizawa, Tomohito Masaki,
Masatoshi Shibuya, Naoki Kanoh, and Yoshiharu Iwabuchi*
Graduate School of Pharmaceutical Sciences, Tohoku University, 6-3 Aobayama,
Sendai 980-8578, Japan. E-mail: iwabuchi@mail.pharm.tohoku.ac.jp
Abstract – The enantio- and diastereocontrolled total synthesis of
(–)-salinosporamide A, a potent 20S proteasome inhibitor, was accomplished
through organocatalytic aldolization, diastereoselective Claisen condensation, a
Rh-catalyzed Reformatsky reaction, and an AZADO-catalyzed oxidative
ꢀ-lactonization reaction as the key reactions.
Salinosporamide A (1, aka NPI-0052, marizobnib) was discovered by Fenical and coworkers as a
cytotoxic principle produced by a marine actinomycete Salinosopra tropicana.1
Structurally,
salinosporamide A has the same unique fused ꢀ-lactone-ꢁ-lactam core as omuralide (2), a well-known
20S proteasome inhibitor derived from lactacystin (3),2 but is distinguished by its five contiguous
asymmetric centers consisting of a ꢀ-chloroethyl group, two quaternary carbons, and a 2-cyclohexene
ring. Salinosporamide A has been reported to be approximately 35 times more effective at proteasome
inhibition than omuralide,1 making it as a candidate for the development of anticancer drugs.3 The
beneficial pharmacological profile of salinosporamide A coupled with its structural complexity has
spurred intense effortsto synthesize it.
HO2C
Cl
O
O
O
NHAc
NH
O
S
NH
NH
O
OH
OH
O
HO
O
O
OH
lactacystin (3)
omuralide (2)
salinosporamide A (1)
Figure 1. Structures of salinosporamide A (1), omuralide (2), and lactacystin (3)

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HETEROCYCLES, Vol. 81, No. 10, 2010
To date, a number of total4,5 and formal6 syntheses have been accomplished, most of which employ
Corey’s approach4a to realize the diastereoselective installation of the cyclohexene ring of 1 in the later
stage of the synthesis, with the exception of the Nagamitsu-Omura synthesis,4e in which the cyclohexene
moiety is mounted in the early stage. We now report the enantio- and diastereocontrolled synthesis of
(–)-salinosporamide A (1) featuring the extensive adoption of aldol and related reactions, where the
cyclohexene ring is installed in the early stage of the synthesis.
Our retrosynthetic analysis of 1 is illustrated in Scheme 1. We set 4 as a logical subgoal, of which the
deprotected oxazoline and the methyl ester moieties could serve as suitable intermediates to obtain 1.
Among several possible disconnections, we chose the intermolecular aldol reaction of the ketone 5 and an
enolate derived from ꢀ-butyrolactone, which would ultimately allow us to secure the vicinal asymmetric
quaternary centers in a diastereocontrolled manner. The quaternary center of the ketone 5 should be
secured from the oxazoline 6 via a diastereocontrolled mixed Claisen condensation with an acetyl
counterpart. The precursor of the oxazoline 6 was expediently assigned to the known aldol 7:7 the
highly diastereocontrolled synthesis of the enantiomeric ent-7 was previously attained via an
L-proline-catalyzed aldol reaction of (S)-Garner’s aldehyde8a and cyclohexanone.
OM
Cl
Ar
O
O
Ar
O
O
N
O
NH
O
OH
N
O
O
O
CO2Me
O
OH CO2Me
5
4
1
aldol or Reformatsky
Claisen
X
Ar
Boc
Boc
N
N
OH
CO2H
N
H
O
N
O
O
CHO
O
O
MeO2C
ref. 7
(R)-Garner's
aldehyde
7
6
aldol
Scheme 1. Retrosynthetic analysis of salinosporamide A (1)
The oxazoline 12, serving as the stereochemical basis throughout the synthesis, was prepared as shown in
Scheme 2. In accordance with the procedure developed by Rode,7 (R)-Garner’s aldehyde8b was treated
with cyclohexanone in the presence of 20 mol% D-proline in CHCl3-DMSO (3:1) in 5 °C to give 7 with
81% yield and a high diaseteroselectivity of over 98% de, of which the diastereochemical preference is

HETEROCYCLES, Vol. 81, No. 10, 2010
2241
rationalized by the transition state model A. 7 Upon TMS etherification, triflation of the kinetically
generated enolate of 7 afforded 8. Pd-catalyzed reduction of the triflate 8 employing formic acid gave 9
with concomitant desilylation. The exposure of 9 to a mixture of cHCl and AcOEt effected the
deprotection of the acetonide and the Boc group to give the corresponding amino-diol, which was then
condensed with p-chlorobenzoyl chloride to give 10. Note that the installation of the p-chlorobenzoyl
group was not the result of a capricious choice (vide infra).9 Treatment of 10 with a catalytic amount of
TEMPO in the presence of PhI(OAc)210 in CH3CN-H2O (1:1) led to the selective oxidation of the primary
alcohol moiety, which upon treatment with diazomethane gave the methyl ester 11. The cyclization of
11 to the oxazoline 12 was accomplished in boiling toluene using a catalytic amount of p-TsOH and
Dean-Stark apparatus.
O
Boc
Boc
N
N
OH
H
O
O
O
N
a
b, c
O
CHO
O
O
H
N
7
(R)-Garner's aldehyde
A
(81%, >98% de)
Boc
Ar
Boc
Boc
O
OH
N
OTMS
N
OH
HN
OTf
d
e, f, g
O
O
HO
8
9
(74% from 7)
10
(88%)
Ar
Ar
O
j
h, i
HN
OH
N
O
Ar =
Cl
MeO2C
MeO2C
11
(78%)
12
(47%, BRSM 89%)
Scheme 2. Synthesis of oxazoline 12, equivalent to 6. Reagents and conditions: (a) D-proline (0.2
equiv), cyclohexanone (2 equiv), CHCl3-DMSO (3:1), 5 °C, 3 d; (b) TMSCl (2 equiv), Et3N (4 equiv),
DMAP (0.1 equiv), CH2Cl2, 0 °C, 1.5 h; (c) KHMDS (1.3 equiv), PhNTf2 (1.1 equiv), THF, –78 to
–40 °C, 1 h; (d) Pd(OAc)2 (0.05 equiv), PPh3 (0.1 equiv), HCO2H (2.5 equiv), n-Bu3N (3 equiv), DMF,
60 °C, 12 h; (e) cHCl, AcOEt, rt, 0.5 h; (f) p-chlorobenzoyl chloride (1.5 equiv), Et3N (4 equiv), DMAP
(0.03 equiv), CH2Cl2, rt, 0.5 h; (g) K2CO3 (10 equiv), MeOH, rt, 2 h; (h) TEMPO (0.25 equiv), PhI(OAc)2
(2.6 equiv), MeCN-H2O (1:1), 0 °C, 15 h; (i) CH2N2 in Et2O, MeOH, 0 °C; (j) p-TsOH (0.05 equiv),
toluene, Dean-Stark, reflux, 12 h.

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HETEROCYCLES, Vol. 81, No. 10, 2010
Having formed the projected chiral platform 12, we considered the formation of the vicinal quaternary
centers in 1 (Scheme 3). The deprotonation of 12 with lithium diisopropylamide in THF and acylation
of the resulting enolate with acetyl chloride afforded the required 13 exclusively with 76% yield. The
next step, the attachment of ꢀ-butyrolactone, turned out to be troublesome: all the attempted aldol
reactions failed to give even a trace amount of the anticipated product and instead gave
2-acetyl-ꢀ-butyrolactone (14) as a major product, indicating the formation of the primary adduct B and the
prompt collapse via a retro-Claisen reaction. After many unsuccessful trials, we decided to make a
detour so as to accommodate the transient oxy-anion species generated by the nucleophilic attack of the
enolate counterpart (Scheme 3).
Ar
Ar
O
O
O
N
N
O
O
a
b
12
O
O
O
CO2Me
O
CO2Me
B
13 (76%)
14
Ar
Ar
Ar
N
O
N
O
N
O
i
c
h
d, e, f, g
13
HO
O
HO
CO2Me
OAc
OAc
16 (95% from 13)
15
17 (91%)
HO
O
O
Ar
HO
O
Ar
O
O
O
O
Ar
N
l
k
N
O
N
NH
OH
+
O
7
HO
8
HO
HO
HO
AcO
OAc
18a (31%)
OAc
AcO
18b (30%)
(87%)
19
20
(49% from 18b)
j
Scheme 3. Synthesis of ꢀ-lactam 20. Reagents and conditions (Ar = p-Cl-C6H4-): (a) n-BuLi (2 equiv),
i-Pr2NH (2.2 equiv), THF, –78 °C; AcCl (3 equiv), 15 min; (b) ꢀ-butyrolactone, LDA, THF, –78 °C; (c)
NaBH4 (1.5 equiv), MeOH, –40 °C, 2 h; (d) DHP (5 equiv), PPTS (0.22 equiv), CH2Cl2, reflux, 3 h; (e)
DIBALH (3 equiv), THF, –5 °C, 9 h; (f) Ac2O (3 equiv), Et3N (6 equiv), DMAP (0.1 equiv), CH2Cl2,
reflux, 3 h; (g) Me2AlCl in hexane (2 equiv), CH2Cl2, 0 °C to rt, 0.5 h; (h) 1-Me-AZADO (0.11 equiv),
PhI(OAc)2 (1.65 equiv), CH2Cl2, rt, 12 h; (i) (Ph3P)3RhCl (0.05 equiv), ꢁ-bromo-ꢀ-butyrolactone (4
equiv), Et2Zn (4 equiv), THF, –20 °C, 3 h; (j) DBU (1.1 equiv), CH2Cl2, 0 °C, 12 h; (k) NaBH3CN (5
equiv), AcOH-THF (2:1), 0 °C, 6 d; (l) 1,2-propanedithiol (4.2 equiv), cHCl, CF3CH2OH, rt, 0.5 h.

HETEROCYCLES, Vol. 81, No. 10, 2010
2243
To this end, ketone 13 was selectively reduced to the alcohol 15 by treatment with NaBH4 in MeOH at
–40 °C. Upon THP protection, the reduction of the ester moiety with DIBALH, acetylation, and the
removal of the THP group, we obtained 16 from 15. Oxidation of the highly congested alcohol 16 was
easily achieved under 1-Me-AZADO11-catalyzed conditions using PhI(OAc)2 to give 17 as a revised
substrate for attaching the ꢀ-butyrolactone moiety. Owing to the challenging steric hindrance of 17,
most
of
attempted
aldolizations
including
the
Mukaiyama
aldolization12
using
(4,5-dihydrofuran-2-yloxy)trimethylsilane were unsuccessful and only Shibasaki’s protocol13 using
CuF·3PPh3·2EtOH/(EtO)3SiF gave the desired adduct with 8% yield. Eventually, we found that the
Honda variant14 of the Reformatsky conditions gave the best results. Thus, 17 was treated with
ꢁ-bromo-ꢀ-butyrolactone and Et2Zn in the presence of 5 mol% Wilkinson’s catalyst to give 18a and 18b
in 31% and 30% isolated yields, respectively. The relative stereochemistry of 18a was confirmed by
X-ray analysis of the corresponding ꢂ7,8-hydrogenated derivative.15 Fortunately, 18a was cleanly
isomerized to the desired epimer 18b upon treatment with DBU.
Detachment of the
chlorophenylmethylene portion of 18b was best effected in a two-step sequence through the convenient
formation of ꢀ-lactam: (1) reduction of oxazoline with NaBH3CN4a to form 19 via an intramolecular
translactamization, (2) removal of the p-chlorobenzylidene group with cHCl and 1,3-propanedithiol16 to
give 20. It should be stressed that the p-methoxy counterpart4a of 18b was significantly resistant9 to
reduction, resulting in a significant yield loss (Scheme 3).
To progress from 20 to (–)-1 (Scheme 4), the primary and secondary hydroxyl groups had selectively to
be protected in a discriminable manner, with the acetate group detached, of which purpose was
expediently attained via the selective TBDPS etherification, deacetylation, selective TMS etherification of
the primary and secondary hydroxyl groups, and selective removal of the primary TMS ether on treatment
with silica gel to furnish 21. Fortunately, upon the attempted oxidation of the primary alcohol under
AZADO11-catalyzed conditions using PhI(OAc)2, 21 directly afforded ꢃ-lactone 22. Note that the use of
TEMPO17 significantly retarded the formation of ꢃ-lactone. Global deprotection of the silyl groups in 22,
followed by treatment of the resulting diol with Ph3PCl24a finally gave (–)-salinosporamide A. The
specific rotation, melting point, and spectroscopic properties of the synthetic 1 were full accordance with
the reported data.
In conclusion, we have developed an enantio- and diastereocontrolled route to synthesize
(–)-salinosporamide A from (R)-Garner aldehyde. The key features of the synthetic route are the
Rh-catalyzed Reformatsky reaction of 17 to 18, and the simple, one-step, oxidative ꢃ-lactonization of 22
catalyzed by AZADO. Additional information on the chemo- and diastereoselective manipulations

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HETEROCYCLES, Vol. 81, No. 10, 2010
around the ꢀ-lactone-ꢁ-lactam motif should provide useful insight for the design and synthesis of
advanced 20S proteasome inhibitors related to salinosporamide A/lactacystin.
TBDPSO
TBDPSO
O
O
NH
NH
O
OTMS
OTMS
d
e
a, b, c
20
HO
O
HO
21 (98%)
22 (78%)
Cl
HO
O
O
NH
O
NH
O
OH
OH
f
[ꢀ]D27 –72.5 (c 0.17, CHCl3)
[ꢀ]D25 –72.9 (c 0.55, CHCl3)
O
O
lit.1
,
23
1 (75% from 22)
Scheme 4. Total synthesis of salinosporamide A (1). Reagents and conditions: (a) TBDPSCl (10 equiv),
Et3N (30 equiv), DMAP (2 equiv), CH2Cl2, rt, 12 h; (b) Amberlyst-26, MeOH, rt, 2.5 h; (c) TMSCl (5
equiv), Et3N (10 equiv), DMAP (0.5 equiv), CH2Cl2, rt, 12 h; SiO2, CHCl3-AcOEt, 15 min; (d) AZADO
(0.07 equiv), PhI(OAc)2 (4 equiv), CH2Cl2, rt, 4 h; (e) aq. HF, MeCN, rt, 1.5 h; (f) Ph3PCl2 (17 equiv),
pyridine-MeCN (1:1), rt, 2 h.
ACKNOWLEDGEMENTS
This work was financially supported in part by a Grant-in-Aid for Scientific Research on Priority Areas
180320110 and Grant-in-Aid for Global COE Program for “International Center of Research ꢀ Education
for Molecular Complex Chemistry” from Ministry of Education, Culture, Sports, Science, and
Technology, Japan.
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The relative stereochemistry of ꢀ7,8-H2-18b was

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HETEROCYCLES, Vol. 81, No. 10, 2010
confirmed by observing the n.O.e of ꢀ7,8-H2-19.
Cl
Cl
5 mol% (Ph3P)3RhCl
O
Br
O
O
O
N
O
N
O
7
4 equiv Et2Zn
THF, 0 °C, 3 h
8
O
HO
OAc
OAc
ꢀ7,8-H2-17
ꢀ7,8-H2-18a (37%)
+
ORTEP of ꢀ7,8-H2-18a
ꢀ7,8-H2-18b (32%)
Cl
Cl
HO
HO
O
Cl
5 equiv NaBH3CN
O
N
OAc
O
O
HO
N
AcOH-THF (3:1)
0 °C, 6 d
N
O
O
H
O
HO
AcO
H3C
Cy
H
HO
n.O.e
OAc
ꢀ7,8-H2-18b
ꢀ7,8-H2-19 (66%)
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