Synthesis and evaluation of the antitumor agent TMC-69-6H and a focused library of analogs

Article abstract of DOI:10.1016/j.tet.2004.06.139

A concise, efficient and flexible total synthesis of the potent antitumor agent TMC-69-6H (2) is described. Key steps involve the palladium catalyzed regioselective addition of 4-hydroxy-2-pyridone 5 to pyranyl acetate 6 which is accompanied by a spontaneous 1,4-addition of the phenolic -OH group to the emerging enone to give the tricyclic product 7 in excellent yield. When this reaction is carried out with optically enriched (S)-6 (conveniently prepared by a lipase catalyzed kinetic dynamic resolution) in the presence of the chiral ligand (S,S)-12 and allylpalladium chloride dimer, the ensuing matched situation delivers the key building block (-)-7 in 96% ee. Its further elaboration into 2 involves a Julia-Kocienski olefination with tetrazolylsulfone 19 and a final N-oxidation effected by the peroxomolybdenum complex [(pyridine)MoO ₅(HMPA)] to form the hydroxamic acid motif. The flexibility inherent to this route allows for the preparation of a focused library of analogues for biochemical evaluation. The results obtained show that N-hydroxy-2-pyridone derivatives constitute a promising new class of selective phosphatase inhibitors. In contrast to previous reports in the literature, however, TMC-69-6H and congeners are found to exhibit pronounced activities against the tyrosine protein phosphatase PTB1B, the dual specific phosphatase VHR, and the serine/threonine phosphatase PP1, while being only weak inhibitors for the dual specific phosphatases Cdc25 A and B. Two key intermediates of the synthesis route have been characterized by X-ray crystallography. Graphical Abstract.

Full text of DOI:10.1016/j.tet.2004.06.139

Tetrahedron 60 (2004) 9543–9558
Synthesis and evaluation of the antitumor agent TMC-69-6H
and a focused library of analogs
a,
Alois Furstner, Fabian Feyen, Heino Prinz and Herbert Waldmann
a
b
b
*
¨
a
¨ ¨
Max-Planck-Institut fur Kohlenforschung, Kaiser-Wilhelm-Platz 1, D-45470 Mulheim/Ruhr, Germany
b
¨
Max-Planck-Institut fur Molekulare Physiologie, D-44227 Dortmund, Germany
Received 26 April 2004; revised 1 June 2004; accepted 3 June 2004
Available online 21 August 2004
Abstract—A concise, efficient and flexible total synthesis of the potent antitumor agent TMC-69-6H (2) is described. Key steps involve the
palladium catalyzed regioselective addition of 4-hydroxy-2-pyridone 5 to pyranyl acetate 6 which is accompanied by a spontaneous 1,4-
addition of the phenolic –OH group to the emerging enone to give the tricyclic product 7 in excellent yield. When this reaction is carried out
with optically enriched (S)-6 (conveniently prepared by a lipase catalyzed kinetic dynamic resolution) in the presence of the chiral ligand
(S,S)-12 and allylpalladium chloride dimer, the ensuing matched situation delivers the key building block (K)-7 in 96% ee. Its further
elaboration into 2 involves a Julia–Kocienski olefination with tetrazolylsulfone 19 and a final N-oxidation effected by the
peroxomolybdenum complex [(pyridine)MoO5(HMPA)] to form the hydroxamic acid motif. The flexibility inherent to this route allows
for the preparation of a focused library of analogues for biochemical evaluation. The results obtained show that N-hydroxy-2-pyridone
derivatives constitute a promising new class of selective phosphatase inhibitors. In contrast to previous reports in the literature, however,
TMC-69-6H and congeners are found to exhibit pronounced activities against the tyrosine protein phosphatase PTB1B, the dual specific
phosphatase VHR, and the serine/threonine phosphatase PP1, while being only weak inhibitors for the dual specific phosphatases Cdc25 A
and B. Two key intermediates of the synthesis route have been characterized by X-ray crystallography.
q 2004 Elsevier Ltd. All rights reserved.
1. Introduction
crucial regulatory functions at the crossroads between
cellular proliferation, cell cycle arrest, and apoptosis.
Their oncogenic properties together with the fact that
Cdc25A and B are overexpressed in many human tumors
render these isoenzymes molecular targets of utmost interest
in the quest for anticancer drugs.7–9
Reversible phosphorylations of proteins mediate innumer-
able biological processes, and aberrant phosphorylation can
cause the development of human diseases such as cancer
and diabetes.1 As a consequence, all enzymes involved in
the regulation of protein phosphorylation sates represent
potential targets for current medicinal chemistry and
chemical biology research. While the phosphorylating
protein kinases have already been intensively studied,2 it
was only recently that their natural antagonists, the protein
phosphatases (PPs) responsible for the catalyzed hydrolysis
of phosphate esters on tyrosine, serine and threonine,
received similar attention.3–7
Among them, the dual specific protein phosphatases (PP’s)
of the Cdc25 family are particularly attractive for their
eminent role in cell cycle control.8 Their physiological
substrates are cyclin-dependent kinases which, in turn,
trigger key transitions in the process of eukaryotic cellular
division. Hence, the homologous Cdc25 enzymes exert
The number of small molecules that qualify as lead
structures in the search for selective inhibitors of Cdc25
phosphatases, however, is rather limited. A particularly
promising candidate is the N-hydroxy-2-pyridone derivative
TMC-69 (1) isolated from the culture broth of the fungus
Keywords: Heterocycles; Julia–Kocienski olefination; N-oxidation; Palla-
dium; Phosphate inhibitors.
* Corresponding author. Tel.: C49-208-306-2342; fax: C49-208-306-
2994; e-mail: fuerstner@mpi-muelheim.mpg.de
0040–4020/$ - see front matter q 2004 Elsevier Ltd. All rights reserved.
doi:10.1016/j.tet.2004.06.139

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A. Furstner et al. / Tetrahedron 60 (2004) 9543–9558
9544
Chrysosporium sp. TC 1068.10 According to the literature,
this compound is distinguished by IC50 values in the low
micromolar range as well as by a surprising selectivity for
Cdc25 A and B over other phosphatases. While 1 itself is
inherently labile and degrades within 2 weeks even at 0 8C,
hydrogenation of its triene moiety results in a significantly
improved stability and was reported to enhance the
inhibitory activity as well as the cytotoxicity even further.
Importantly, TMC-69-6H (2) thus formed has also proven
effective in vivo for the treatment of P388 murine leukemia
and B16 melanoma in nude mice, leading to an increase on
life span of up to 105.9% at a dose of 1.25 mg/kg i.p.10
Due to this favorable profile, TMC-69-6H was selected for
further study in our program directed at the identification
and development of novel classes of phosphatase inhibi-
tors.11–13 Outlined below is the first total synthesis of both
the (17R)- and the (17S)-configured isomers of 2, since the
absolute stereochemistry at this remote chiral center in the
natural product 1 and its derivative 2 has not yet been
established.14 The chosen route is based on a novel
palladium catalyzed fragment coupling process for the
assembly of the core structure. Moreover, its inherent
flexibility allowed for the synthesis of a focused library of
analogues which enabled first insights into structure/activity
relationships (SAR). Surprisingly though, the consistent set
of biochemical data obtained with these samples is not in
accord with the claims previously made in the literature.10
Rather than being potent inhibitors of Cdc25A and B, we
find pyridone derivatives of this series to inhibit the tyrosine
protein phosphatase PTB1B, the dual specific phosphatase
VHR, and the serine/threonine phosphatase PP1 much more
effectively. The relevance of this finding, which results in
the re-definition of the activity profile of TMC-69-6H, is
outlined below.
Scheme 1. Retrosynthetic analysis of TMC-69-6H (2).
as granted because 2-pyridones tend to react with allylic
substrates at their N-atom in the presence of palladium
catalysts,17 and palladium catalyzed transformations of
6-hydroxy-6H-pyran-3-one E (XZOR) derivatives are
rather scarce and seem to be restricted to reactions with
O-nucleophiles.18 To the best of our knowledge, no Pd-
catalyzed C-arylation of such a compound with a phenol
derivative has previously been described. If successful,
however, the envisaged fragment coupling process would
greatly contribute to the overall flexibility of the synthesis
route and add a new facet to organopalladium chemistry in
general.19,20
2. Results and discussion
2.2. Total synthesis
2.1. Retrosynthetic analysis
The required 4-hydroxypyridone 5 was prepared on a
large scale by condensation of malonic acid dichloride 3
with 2-phenylacetonitrile followed by hydrogenolytic
removal of the residual chloride in 4 (Scheme 2).21 We
were pleased to see that compound 5 reacted smoothly and
regioselectively at its ‘enolic’ site with rac-622 in the
presence of Pd(PPh3)4 cat. and Et3N in DMF to give the
tricyclic product (G)-7 in 89% yield. The palladium-
catalyzed C–C-bond formation is accompanied by a
spontaneous 1,4-addition of the 4-OH group to the enone
entity of the emerging product; we are unaware of any
precedence for this transformation. Attempts to perform this
reaction enantioselectively using rac-6 as the substrate and
the chiral diphosphine 1223 as ligand to Pd were unreward-
ing (ee w30%).
Since the hydroxamic acid function likely contributes to the
lability of 2, it was decided to install this group at a late
stage of the synthesis by N-oxidation of the pyridone
precursor A (Scheme 1). The alkyl side chain of the latter
can be attached to the heterocyclic core by an olefination/
hydrogenation sequence which provides opportunities for
structural variations of this part of the molecule at a later
stage. The required precursor B featuring two trans-
disposed substituents might be secured by a 1,4-addition
of a suitable methyl donor to the corresponding enone C
which should derive from pyridone D and the pyranone E.
Although one might envisage to join these building blocks
by established oxo-carbenium cation chemistry,15 recourse
to palladium catalyzed C–C-bond formations should allow
to control the absolute stereochemistry at the ring junction
and therefore provide a more attractive solution.16 To this
end, however, the metal template has to force the ambident
pyridone D to act exclusively as a C- (rather than O- or N-)
nucleophile and its equally ambident reaction partner E to
behave solely as an allylcation equivalent rather than as a
Michael acceptor. Such a reactivity pattern cannot be taken

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A. Furstner et al. / Tetrahedron 60 (2004) 9543–9558
9545
spectra.21,25 While chromophors of the general type F have
an absorption maximum in the range of lmaxZ245–247 nm,
their constitutional isomers G absorb at lmaxZ233–235 nm.
Since the tricyclic product formed in the palladium-
catalyzed fragment coupling process shows a lmax at
245 nm, we ascribe the 2(1H)-pyridinone structure 7 to
this key building block.
The cyclic ether moiety in 7 acts as a temporary protecting
group for the enone, which can be released by a retro-
Michael reaction on treatment with oxophilic reagents in the
presence of a base.26 Optimal results were obtained with
TBSCl in combination with Et3N. The bis-TBS-ether 8,
though labile, can be isolated in pure form, whereas its
TMS-counterpart is too unstable to be of practical use. The
structure of this compound in the solid state is shown in
Figure 1. As can be seen, one p-face of the enone (C7A–
C8A) is strongly shielded by the bulky silyl group attached
to O4A. Although it has not been investigated if this product
adopts a similar conformation in solution, this particular
steric arrangement augured for a good selectivity in the
subsequent 1,4-addition reaction.
Scheme 2. Synthesis of (17S)-2: [a] phenylacetonitrile, 4d, 50%; [b] H2
(1 atm), Pd/C, EtOH, 60 8C, 97%; [c] [(allyl)PdCl]2 (0.5%), ligand 12
(1.5%), DMF, 65% (eeZ96%); [d] TBSCl, Et3N, 18 h, 78%; [e] Me2CuLi,
THF, K70 8C; [f] LiHMDS, (S)-19, THF, K78 8C, 69% (over both steps);
[g] H2, Pd/C, EtOH; [h] TBAF, THF, 69% (over both steps); [i] (i)
HN(SiMe3)2, TMSCl cat., reflux; (ii) (pyridine)MoO5(HMPA), CH2Cl2;
(iii) sat. aq. EDTA-Na, EtOAc, 71%.
In fact, exposure of compound 8 to Me2CuLi afforded the
somewhat sensitive trans-disubstituted ketone 9 in good
yield and excellent diastereoselectivity (de O90%).
Crystals suitable for X-ray analysis could be obtained
from the closely related compound 25 carrying a chloride
substituent on the pyridone ring (see below). Its structure in
the solid state is shown in Figure 2.
Gratifyingly though, the use of 12 in combination with
enantiomerically enriched (S)-6 (eeZ81%), which is easily
prepared on a multigram scale by a lipase catalyzed
dynamic resolution,24 served our purpose very well,
delivering the tricyclic ketone (K)-7 in good yield (65%)
and excellent optical purity (eeZ96%) (Scheme 2). Thereby
it is essential to keep the catalyst loading low to avoid
partial racemization of the intermediates via ligand transfer
processes.
Although ketone 9 can be isolated in pure form, it turned out
to be rather unstable. Therefore the crude product was
immediately subjected to a Julia–Kocienski olefination
reaction for the introduction of the aliphatic side chain.27,28
The required sulfones were prepared from citronellene 14 as
shown in Scheme 3 for (R)-19. Selective ozonolysis of the
more highly substituted double bond29,30 followed by
routine elaboration of the resulting aldehyde 15 gave
alcohol 17, which condensed with commercial 1-phenyl-
1H-tetrazol-5-thiol under Mitsunobu conditions31 to give
sulfide 18. Oxidation with aqueous H2O2 in the presence of
catalytic amounts of ammonium molybdate furnished
sulfone (R)-19 in good overall yield. Since both isomers
of citronellene are commercially available, the antipodal
sulfone (S)-19 is equally accessible by this route.
It is worth mentioning that an unambiguous structure
assignment of this tricyclic product is not trivial. Specifi-
cally, one has to consider that the 2-hydroxy-4-pyridone
tautomer of 5 might come into play and lead to the
formation of compound 13, which is difficult to distinguish
from 7 by NMR and IR. Recent studies, however, revealed
that such tautomeric ethers show distinctly different UV

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A. Furstner et al. / Tetrahedron 60 (2004) 9543–9558
9546
Figure 1. Molecular structure of compound 8 in the solid state. Anisotropic displacement parameters are shown at the 50% probability level, hydrogen atoms
˚
are omitted for clarity. Only one of the two independent molecules in the unit cell is depicted. Selected bond length (A) and angles (8): C(1A)–N(1A) 1.341(6),
C(1A)–C(2A) 1.371(7), C(2A)–C(3A) 1.409(7), C(3A)–C(4A) 1.383(6), C(4A)–C(5A) 1.401(6), C(5A)–N(1A) 1.330(6), C(6A)–C(7A) 1.488(7), C(7A)–
C(8A) 1.352(7), C(8A)–C(9A) 1.439(8), C(9A)–O(2A) 1.229(7), C(9A)–C(10A) 1.488(9), C(10A)–O(1A) 1.427(6), C(6A)–O(1A) 1.435(5), C(3A)–O(3A)
1.375(6), C(5A)–O(4A) 1.350(5), C(8A)–C(7A)–C(6A) 120.9(5), C(7A)–C(8A)–C(9A) 120.3(6), O(2A)–C(9A)–C(8A) 122.8(7), O(1A)–C(6A)–C(7A)
111.5(4), O(1A)–C(10A)–C(9A) 113.5(5).
Reaction of the lithiated sulfone (S)-19 with the crude
ketone 9 provided alkene 10 as a 1:1 mixture of isomers.
HPLC analyses on chiral columns and comparison with the
racemic series confirmed that no racemization had occurred
during this or any of the preceding steps (eeZ97%).
Subsequent hydrogenation of the (E,Z)-mixture of 10 over
Pd/C followed by cleavage of the –OTBS groups furnished a
separable 5:1 mixture of the desired product 11 and its C-10
diastereomer in 69% yield over both steps. The axial
orientation of the alkyl chain on the tetrahydropyran ring in
11 was evident from an analysis of the pertinent coupling
constants and NOESY data (Scheme 4).
treated with the peroxomolybdenum complex
[(pyridine)MoO5(HMPA)].34 An aqueous work-up of the
reaction mixture with EDTA to sequester all metal cations
completed the first total synthesis of (17S)-TMC-69-6H
(17S-2).
As mentioned earlier, no secured information concerning
the absolute stereochemistry of the remote chiral center on
the lateral chain of compound 2 derived from natural
sources is presently available.10 Therefore the same
For the final N-oxidation to the desired hydroxamic acid
derivative,33 compound 11 was refluxed with hexamethyl-
disilazane (HMDS) and the resulting bis-silylether was
Figure 2. Molecular structure of compound 25 in the solid state. Anisotropic displacement parameters are shown at the 50% probability level, hydrogen atoms
˚
are omitted for clarity. Selected bond length (A) and angles (8): N(1)–C(2) 1.335(6), N(1)–C(6) 1.342(6), C(2)–C(3) 1.385(7), C(3)–C(4) 1.408(7), C(4)–C(5)
1.395(7), C(5)–C(6) 1.398(7), O(4)–C(4) 1.361(6), O(6)–C(6) 1.342(6), C(51)–C(56) 1.527(7), C(55)–C(56) 1.533(7), C(54)–C(55) 1.496(8), C(54)–O(58)
1.201(6), C(53)–C(54) 1.504(8), O(52)–C(53) 1.420(6), C(51)–O(52) 1.445(6), C(57)–C(56)–C(51) 110.6(4), C(51)–C(56)–C(55) 108.9(4), C(5)–C(51)–
C(56) 113.8(4), O(52)–C(51)–C(5) 106.3(4).

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A. Furstner et al. / Tetrahedron 60 (2004) 9543–9558
9547
analogues for biological evaluation in order to gain first
insights into structure/activity relationships.
The ketone group in 9 provides an opportunity for late stage
divergent modification that might allow to probe the
importance of the aliphatic tail for binding to the protein.
To this end, compound 9 was subjected to Julia–Kocienski
olefination reactions with the two unbranched alkyl sulfones
20a,b.27,36 The olefins 21a,b thus formed were hydrogen-
ated over Pd/C and the major stereoisomers of the resulting
products (d.r.Z5:1) were separated and N-oxidized with
[(pyridine)MoO5(HMPA)] as outlined above to give
products 23a,b, respectively, differing from the lead
compound 2 only in their lipophilicity (Scheme 5).
Next, the synthesis was repeated with the 6-chloropyridone
derivative 421 which was formed as the primary product in
the condensation reaction shown in Scheme 2. Gratifyingly,
the aryl halide did neither interfere in the palladium-
catalyzed fragment coupling step to give optically active
tricyclic ketone 24 (eeZ95%) nor in the cuprate addition
reaction introducing the methyl branch. However, dehalo-
genation competes with the hydrogenation of the double
bond in product 26; therefore careful monitoring of the
reaction was necessary to obtain product 27 in 44% yield,
which was oxidized according to the established protocol to
give the desired analogue 28 for biological evaluation
(Scheme 6).
Scheme 3. [a] O3, CH2Cl2, K78 8C, then Me2S, 98%; [b] (EtO)2P(O)CH2-
COOMe, NaH, THF, K78 8C, 87%; [c] Dibal-H, Et2O, 91%; [d] H2, Pd/C,
88%; [e] 1-phenyl-1H-tetrazole-5-thiol, DEAD, PPh3, THF, 68%; [f]
(NH4)6Mo7O24$4H2O (0.1 equiv.), H2O2 (10 equiv.), EtOH, 93%.
Finally, more substantial structural modifications were
made by substituting the entire lipophilic segment of the
natural product by bare alkyl chains attached to C-3 of the
pyridone (Scheme 7). For this purpose, we investigated if
simple allylic esters undergo similar palladium catalyzed
cross coupling reactions with pyridone 5 as does the more
activated substrate 6 used in the total synthesis of 2. In fact,
acetates 29a,b afforded the desired C-allylated products
30a,b, although more forcing conditions were required and
the yields were lower than those obtained with 6.
Subsequent hydrogenation followed by N-oxidation pro-
ceeded uneventfully and afforded product 32 as a simplified
analogue of TMC-69-6H.
sequence of reactions was repeated using ketone (K)-9 and
the antipodal sulfone (R)-19 to give (17R)-2 in similar
overall yield. The diastereomeric compounds (17R)-2 and
(17S)-2 thus obtained, however, are virtually indistinguish-
able by NMR and match the literature data reported for
TMC-69-6H very well. Even a direct comparison of their
spectra recorded at 600 MHz with those of an authentic
sample does not allow us to rigorously assign the absolute
stereochemistry of TMC-69-6H at that distal site (Tables 1
and 2).35
2.3. Analogues
2.4. Biochemical investigations: Re-assessment of the
phosphatase inhibitory activity of TMC-69-6H and
analogues
In view of the promising biochemical and biological
properties of TMC-69 and TMC-69-6H described in the
literature,10 the preparation of analogues was called for.
Taking advantage of the flexibility inherent to the synthesis
route described above we prepared a small collection of
The excellent match between the analytical and spectro-
scopic data of (17R)-2 and (17S)-2 prepared by total
synthesis with those reported in the literature leaves no
doubt about the structural integrity of these compounds.
Therefore we were surprised to find that their phosphatase
inhibitory activity significantly deviates from the reported
profile both in terms of potency and selectivity. In contrast
to what has been claimed,10 the compounds prepared by
total synthesis as well as the authentic sample of 2 all turned
out to be only rather weak inhibitors for Cdc25A (IC50O
30 mM) in our assay. Instead, they exhibit promising
activities against the tyrosine protein phosphatase PTB1B,
the dual specific phosphatase VHR, and the serine/threonine
phosphatase PP1 (Table 3).
Scheme 4. Schematic representation of compound 11 with characteristic
NOESY data. The following coupling constants indicate a chair
3
3
conformation of the tetrahydropyran ring: JH7,H8Z10.4 Hz, JH10,H11ax-
Z2.5 Hz, 3JH10,H11eq%1 Hz (TMC-69-6H numbering).32
PTP1B is a key negative regulator of insulin-receptor

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A. Furstner et al. / Tetrahedron 60 (2004) 9543–9558
9548
Table 1. Comparison of the 13C NMR data of (17R)-2 and (17S)-2 in CDCl3 with those of the authentic material published in the literature;10 the accuracy of
the recorded data is G0.1 ppm. Arbitrary numbering scheme as shown in the insert
Position
(17S)-2
(17R)-2
Authentic sample
Literature10
2
3
4
5
6
7
157.2
109.3
160.9
113.5
131.1
81.6
157.2
109.5
161.0
113.7
130.5
81.6
157.4
109.3
161.0
113.7
130.8
81.6
157.0
109.3
160.9
113.5
129.9
81.7
8
9
31.0
36.1
31.0
36.1
31.0
36.1
31.0
36.1
10
11
12
13
14
15
16
17
18
19
20
21
22
23
24
25
33.0
72.4
30.8
30.0
27.1
27.8
36.5
34.4
29.5
11.4
17.8
19.2
133.2
129.2
128.4
127.7
34.0
72.4
30.8
29.9
27.0
27.8
36.5
34.4
29.5
11.4
17.8
19.2
133.1
129.2
128.4
127.7
34.0
72.4
30.8
29.9
27.0
27.8
36.5
34.4
29.5
11.4
17.8
19.2
133.2
129.2
128.4
127.7
34.0
72.5
30.9
30.0
27.1
27.8
36.9
34.4
29.5
11.4
17.9
19.2
133.2
129.2
128.4
127.7
activity, and PTP1B-inhibitors are expected to enhance
insulin sensitivity and act as effective therapeutics for the
treatment of Type II diabetes, insulin resistance and
obesity.6 The vaccinia VH1-related phosphatase VHR is a
physiological regulator of various members of the MAP
kinase family and therefore influences signaling cascades
via the MAP kinase pathway.37 PP1 is a major enkaryotic
phosphatase that regulates diverse cellular processes such as
signal transduction, cell cycle progression, protein syn-
thesis, muscle contraction, carbohydrate metabolism and
transcription. All of these phosphatases have been subject to
intense research activities aimed at the development of
inhibitors for biological studies and drug development.1–7,38
strictly required for phosphatase inhibition, enhances the
potency of such compounds. Secondly, (17S)-2 and (17R)-2
are equally effective inhibitors within the error margins,
thus showing that the configuration at the lateral methyl
branch is hardly relevant. In line with this notion, analogues
23a,b bearing straight-chain alkyl residues were both active,
with the more lipophilic compound 23b being significantly
more potent. Furthermore, alkene groups embedded in the
alkyl side chain are tolerated and their configuration is of
minor importance since both isomers of 10 and 30b led to
respectable phosphatase inhibition. The chloride substituent
on the pyridone ring in 28 is well accommodated, indicating
that further variations in the aromatic part of 2 might be
worthwhile. Equally promising is the fact that even more
severe structural changes as those found in compounds 30–
32 do not annihilate the inhibitory capacity for relevant
phosphatases at all.
The evaluation of TMC-69-6H, its analogues and isomers
described above allows us to draw further conclusions. First,
the comparison of the IC50 values for, e.g. (17S)-2 with
those of the precursor amide (17S)-11 indicate that the
presence of the N-OH group in the heterocycle, though not
Although we are unable to reconcile our findings concerning
Table 2. Comparison of pertinent 1H NMR data of (17R)-2 and (17S)-2 in CDCl3 with those of the authentic material published in the literature;10 the full set of
data is compiled in the Section 3; arbitrary numbering scheme as shown in the insert to Table 1
Position
(17S)-2
(17R)-2
Literature10
–OH
6
9.52
7.67
9.52
7.67
9.51
7.67
7
8
4.68 (d, JZ10.5 Hz)
2.06 (m)
4.68 (d, JZ10.5 Hz)
2.07 (m)
4.68 (d, JZ10.5 Hz)
2.08 (m)
9a
10
11a
11b
1.76 (d, JZ12.2 Hz)
1.62 (m)
3.96 (d, JZ11.5 Hz)
3.72 (dd, JZ11.5, 2.3 Hz)
1.76 (d, JZ11.0 Hz)
1.61 (m)
3.96 (d, JZ11.8 Hz)
3.72 (dd, JZ11.6, 2.5 Hz)
1.77 (d, JZ12.0 Hz)
1.63 (m)
3.96 (d, JZ11.6 Hz)
3.72 (dd, JZ11.6, 2.5 Hz)

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9549
Scheme 7. [a] Pd(PPh3)4 (5 mol%), Et3N, DMF, 110 8C, 59% (30a, RZH);
or: Pd(PPh3)4 (7.5 mol%), NaH, DMF, 125 8C, 49% (30b, RZ(CH2)8Me);
[b] H2 (1 atm), Pd/C, EtOH, quant. 60% (RZ(CH2)8Me); [c] (i) HMDS,
TMSCl cat., reflux; (ii) (pyridine)MoO5(HMPA), CH2Cl2; (iii) sat. aq.
EDTA-Na, EtOAc.
Scheme 5. [a] LiHMDS, 20a,b, THF, K78 8C; [b] H2, Pd/C, EtOH; [c]
TBAF, THF, 32% (21a over three steps), 17% (21b, over three steps); [d] (i)
HN(SiMe3)2, TMSCl cat., reflux; (ii) (pyridine)MoO5(HMPA), CH2Cl2;
(iii) sat. aq. EDTA-Na, EtOAc, 80% (23a), 85% (23b).
the physiological activity profile of TMC-69-6H with the
literature reports,10 the excellent reproducibility of our
results, the internal control against authentic 2, the
consistency within the individual series, and the established
validity of our Cdc25-assay11–13 leave no room for
interpretation. More importantly, these data make clear
that N-hydroxy-2-pyridone derivatives constitute a promis-
ing new class of selective phosphatase inhibitors which
allow for substantial structural variations and therefore
constitute relevant lead compounds for further optimiz-
ation.39 Notably, TMC-69-6H incorporates a structural
framework not present in any of the PTP1B- and PP1
inhibitors developed so far. Its proven activity in cellular
assays as well as in vivo10 renders further research activities
in this field promising. In particular it seems lucrative to
investigate the inhibitory profile of other natural products
containing an N-hydroxy-2-pyridone motif similar to the
one found in TMC-69.40
3. Experimental
3.1. General
All reactions were carried out under Argon in flame-dried
glassware using Schlenk techniques. The solvents were
dried by distillation over the indicated drying agents and
were stored and transferred under Argon: CH2Cl2, Et3N,
DMF (CaH2), toluene, THF, DME (Na), MeOH, EtOH
(Mg), Et2O (Mg-anthracene). Flash chromatography: Merck
silica gel (230–400 mesh) using either hexanes/ethyl acetate
or pentanes/diethyl ether in various proportions as the
eluents. NMR: Spectra were recorded on a Bruker DPX 300,
AMX 400, DMX 600 spectrometer in CDCl3 or CD2Cl2 as
indicated. Chemical shifts (d) are given in ppm relative to
the residual peak of CHCl3 or CHDCl2; coupling constant
(J) in Hz. IR: Nicolet FT-7199, wavenumbers in cmK1. MS:
Varian CH-5 (70 eV), HRMS: Finnigan MAT SSQ 7000
(70 eV). Elemental analyses: H. Kolbe, Mu¨lheim. Com-
mercialy available reagents (Aldrich, Fluka, Strem,
Lancaster) were used as received.
Scheme 6. [a] Compound (S)-6, [(allyl)PdCl]2 (0.5%), ligand 12 (1.5%),
DMF, 71% (eeZ95%); [b] TBSCl, Et3N, 1 h, 39%; [c] Me2CuLi, THF,
K70 8C; [d] LiHMDS, 19, THF, K78 8C; [e] TBAF, THF, E/ZZ1:1, 82%
(over three steps); [f] H2, Pd/C, EtOH, 44%, d.r.Z3:1; [g] (i) HN(SiMe3)2,
TMSCl cat., reflux; (ii) (pyridine)MoO5(HMPA), CH2Cl2; (iii) sat. aq.
EDTA-Na, EtOAc, 77%.
3.1.1. 1-Phenyl-3,4b(S),8,8a(R)-tetrahydro-5,9-dioxa-3-
aza-flouren-4,7-dione (7). A solution of pyridone 5
(1.00 g, 5.34 mmol)21 in DMF (25 mL) and Et3N (0.5 mL)
was stirred for 10 min before [(allyl)PdCl]2 (9.8 mg,
0.03 mmol) and (K)-1,2-bis-N-[20-(diphenylphosphino)-

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A. Furstner et al. / Tetrahedron 60 (2004) 9543–9558
9550
Table 3. IC50 values [mM] of authentic and synthetic TMC-69-6H, its immediate precursors and analogues against different phosphatasesa
Compound
Cdc25A
PTP1B
VHR
PP1
Authentic 2
R50
3.2G1.6
7.0G3.5
6G3
(17R)-2
45G23
4G2.0
6G3
8.5G4.5
(17S)-2
32G16
3.5G1.7
5.5G3
8G4
(17S)-11
R50
11G5.5
9G5
30G5
(17S)-10a
R50
23G11
11G5.5
32G16
28
O50
O50
O50
O50
O50
5.9G4.2
11.3G5.6
2.8G1.5
12G6
26G16.5
18.9G14
26G13
38.5G13
27
47G6
23b
23a
22a
19G8
14.1G8.3
3.1G1.4
O50

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A. Furstner et al. / Tetrahedron 60 (2004) 9543–9558
9551
Table 3 (continued)
Compound
Cdc25A
PTP1B
VHR
PP1
47
32
O50
4.7G1.2
6.1G3.3
9.2G4.6
3.1G1.6
31b
O50
O50
9.5G8.6
33G15
30bb
26G13
O50
a
The enzymatic activity was determined by hydrolysis of para-nitrophenyl phosphate in standard buffers for PTP1B, VHR and PP1.
1:1 Mixture of isomers.
b
benzoyl]-1(S),2(S)-diaminocyclohexane (S,S)-12 (55.5 mg,
(dd, JZ15.3, 3.9 Hz, 1H); 13C NMR (400 MHz, CDCl3) d
206.5, 170.2, 161.6, 139.8, 130.2, 129.9, 128.6, 128.4,
109.9, 104.5, 83.0, 73.8, 69.1, 39.3; IR (film) 2869, 1737,
1650, 1613, 1544, 1432, 1211, 1094, 1074, 1025, 957, 900,
772, 702; UV [3max (l)]: 12300 (240 nm), 11600 (219 nm);
MS (EI) m/z (rel. intensity) 317 ([MC], 88), 258 (35), 245
(100), 222 (22), 209 (45), 167 (7), 153 (12), 115 (13), 89
(19), 43 (27); HR-MS (ESI-pos) (C16H12NO4Cl) calcd
317.0455, found 317.0450.
0.08 mmol) were introduced. The resulting mixture was
added dropwise to a solution of acetate (S)-6 (1.12 g,
7.20 mmol, eeZ81%)24 in DMF (15 mL) and stirring was
continued for 25 min before the reaction was quenched with
aq. sat. NH4Cl. A standard extractive work up followed by
flash chromatography of the crude product (EtOAc/EtOH,
20:1CEt3N (0.1%, v/v)) afforded compound 7 as a pale
yellow solid (834 mg, 65%). The enantiomeric excess (eeZ
96%) was determined by HPLC on a chiral column
(Chiralcel OJ-R, : 4.6!150 mm; MeCN/water 20:80,
0.5 mL minK1; 308 K; 3.6 MPa; retention times: (K)-7:
9.435 min, (C)-7: 10.237 min). [a]2D0ZK1858 (cZ1.0,
CHCl3/MeOHZ9:1). 1H NMR (400 MHz, DMSO-d6) d
11.66 (s, 1H), 7.64 (s, 1H), 7.50–7.30 (m, 5H), 5.47 (d, JZ
7.3 Hz, 1H), 5.37 (dt, JZ7.0, 4.0 Hz, 1H), 4.01 (d, JZ
18.1 Hz, 1H), 3.64 (d, JZ18.1 Hz, 1H), 3.19 (dd, JZ16.2,
4.0 Hz, 1H), 2.88 (dd, JZ16.2, 4.0 Hz, 1H); 13C NMR
(100 MHz, DMSO-d6) d 208.5, 167.9, 160.3, 137.8, 132.9,
128.7, 127.5, 127.2, 107.7, 105.5, 82.0, 73.8, 68.5, 39.6; IR
(KBr) 3144, 3055, 2973, 2875, 1739, 1662, 1438, 1199,
1086, 775, 698; UV [3max (l)]: 14200 (245 nm), 11600
(210 nm); MS (EI) m/z (rel. intensity) 283 ([MC], 72), 224
(41), 212 (55), 211 (100), 196 (12), 144 (5), 102 (7), 77 (5);
HR-MS (ESI-pos) (C16H13NO4) calcd 283.084270, found
283.084457.
3.1.3. (R)-6-[2,4-Bis-(tert-butyl-dimethyl-silanyloxy)-5-
phenyl-pyridin-3-yl]-6H-pyran-3-one (8). A solution of
compound 7 (50 mg, 0.18 mmol), TBSCl (140 mg,
0.93 mmol) and Et3N (0.5 mL) was stirred for 18 h at
ambient temperature. The mixture was adsorbed on silica
and the product was purified by flash chromatography
(hexanes/Et2O, 10:1CEt3N (0.5%, v/v)) to give compound
8 as a colorless, moisture-sensitive solid (70 mg, 78%).
[a]2D0ZK8.358 (cZ3.5, THF). 1H NMR (400 MHz,
CD2Cl2) d 7.97 (s, 1H), 7.45–7.34 (m, 5H), 7.17 (dd, JZ
10.4, 1.7 Hz, 1H), 6.20 (dd, JZ10.5, 2.7 Hz, 1H), 5.76 (dd,
JZ4.4, 2.2 Hz, 1H), 4.31 (d, JZ16.4 Hz, 1H), 4.20 (dd, JZ
16.2, 2.1 Hz, 1H), 0.95 (s, 9H), 0.93 (s, 9H), 0.34 (s, 3H),
0.30 (s, 3H), K0.26 (s, 3H), K0.37 (s, 3H); 13C NMR
(100 MHz, CD2Cl2) d 194.2, 162.4, 160.0, 153.4, 149.0,
136.6, 130.0, 129.5, 128.4, 126.7, 124.9, 113.1, 72.8, 68.2,
25.8, 25.6, 18.4, 18.2, K4.0, K4.3, K4.5, K4.9; IR (film)
3063, 2955, 2929, 2885, 2858, 1702, 1582, 1461, 1255,
1066, 976, 843, 787, 701; MS (EI) m/z (rel. intensity) 511
([MC], !1) 496 (3), 456 (13), 454 (100), 424 (12), 396 (4),
368 (3), 338 (7), 294 (2), 199 (1), 156 (5), 129 (3), 73 (19);
HR-MS (ESIpos) (C28H42NO4Si2) calcd 512.265241, found
512.265080 (MCH).
3.1.2. 2-Chloro-1-phenyl,3,4b(S),8,8a(R)-tetrahydro-5,9-
dioxa-3-aza-flouren-4,7-dione (24). Prepared as described
above from pyridone 4 (200 mg, 0.90 mmol) and acetate
(S)-6 (210 mg, 1.34 mmol). Pale yellow powder (205 mg,
71%). eeZ95.3% (Chiralcel OD-R, : 4.6!250 mm;
MeCN/water 35:65, 0.5 mL/min; 298 K; 3.3 MPa; retention
times: (C)-(24): 14.465 min, (K)-(24): 16.119 min).
[a]2D0ZK2228 (cZ0.52, CHCl3). 1H NMR (400 MHz,
CDCl3) d 8.75 (s, 1H), 7.46–7.25 (m, 5H), 5.69 (d, JZ
7.3 Hz, 1H), 5.28 (m, 1H), 4.11 (d, JZ18.1 Hz, 1H), 3.86
(d, JZ18.3 Hz, 1H), 3.06 (dd, JZ16.3, 3.9 Hz, 1H), 2.99
3.1.4. (R)-6-[2,4-Bis-(tert-butyl-dimethyl-silanyloxy)-5-
phenyl-pyridin-3-yl]-(R)-5-methyl-tetrahydro-pyran-3-
one (9). A cooled solution (K78 8C) of enone 8 (140 mg,
0.27 mmol) in THF (4 mL) was slowly added to a solution

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A. Furstner et al. / Tetrahedron 60 (2004) 9543–9558
9552
of Me2CuLi [freshly prepared from CuBr$Me2S (259 mg,
1.26 mmol) and MeLi (1.6 M in Et2O, 1.57 mL) in THF
(8 mL) at 0 8C] and the resulting mixture was stirred for 2 h
at K70 8C. Addition of aq. NH4OH/NH4Cl (pH 8–9)
followed by a standard extractive work up provided crude
9 (205 mg, dr O9:1) which is moisture sensitive and
therefore used without further purification in the next step.
2.80 (dd, JZ13.5, 2.7 Hz, 1H), 2.58 (m, 2H), 2.40, (d, JZ
13.5, 2.6 Hz, 1H), 2.15–0.80 (m, 30H), 1.00 (s, 18H) 0.96 (s,
18H), 0.74, (d, JZ6.7 Hz, 6H), 0.70 (d, JZ6.7 Hz, 6H),
0.37 (s, 6H), 0.32 (s, 6H), 0.06 (s, 6H), K0.65 (s, 6H); 13C
NMR (75 MHz, CD2Cl2) d 162.1, 159.6, 147.2, 137.2,
134.3, 134.0, 130.0, 128.3, 128.1, 127.0, 124.6, 124.2,
115.2, 115.1, 78.0, 74.1, 66.5, 42.7, 36.6, 35.0, 34.5, 34.5,
34.2, 33.4, 30.6, 30.3, 29.8, 29.7, 29.6, 27.3, 27.1, 27.0,
27.0, 26.1, 26.0, 25.9, 25.8, 19.1, 18.4, 18.2, 17.8, 17.5,
11.3, 11.3, K4.1, K4.2, K4.2, K4.3, K4.6, K5.3; IR
(film) 2956, 2928, 2857, 1582, 1455, 1444, 1254, 1136,
1058, 1004, 842, 784, 699; MS (EI) m/z (rel. intensity) 637
([MC], !1), 622 (2), 582 (18), 580 (100), 562 (8), 550 (14),
448 (5), 398 (12), 332 (3), 284 (4), 256 (2), 95 (2), 73 (17),
57 (2), 43 (2); HR-MS (ESI-pos) (C38H65NO3Si2) calcd
638.442476, found 638.442434 (MCH).
1
Characteristic data: H NMR (400 MHz, CDCl3) d 7.94 (s,
1H), 7.42–7.30 (m, 5H), 4.79 (d, JZ10.0 Hz, 1H), 4.22 (dd,
JZ15.9, 1.7 Hz, 1H), 3.97 (d, JZ15.9 Hz, 1H), 3.07 (m,
1H), 2.78 (ddd, JZ16.0, 5.3, 1.5 Hz, 1H), 2.20 (dd, JZ16.0,
11.2 Hz, 1H), 0.98 (s, 9H), 0.94 (s, 9H), 0.86 (m, 3H), 0.36
(s, 3H) 0.35 (s, 3H), K0.01 (s, 3H), K0.61 (s, 3H); 13C
NMR (100 MHz, CDCl3) d 207.9, 161.8, 159.7, 148.0,
136.7, 129.9, 128.3, 127.1, 124.5, 113.6, 76.5, 74.3, 46.5,
33.1, 26.0, 26.0, 18.4, 18.2, 18.2, K3.9, K3.9, K4.4, K4.9;
IR (film) 3061, 2956, 2930, 2895, 2858, 1733, 1582, 1456,
1445, 1256, 1056, 976, 840, 784, 700; MS (EI) m/z (rel.
intensity) 527 ([MC], !1), 512 (4), 470 (100), 442 (5), 412
(22), 340 (13), 314 (21), 258 (5), 207 (2), 157 (2), 73 (21);
HR-MS (ESI-pos) (C29H46NO4Si2) calcd 528.296541,
found 528.296878 (MCH).
3.1.7. 2,4-Bis-(tert-butyl-dimethyl-silanyloxy)-3-[(R)-3-
methyl-5-(pentyliden)-tetrahydro-pyran-2-(R)-yl]-5-
phenyl-pyridine (21a). Prepared analogously using sulfone
20a. 1H NMR (400 MHz, CD2Cl2) d 7.88 (s, 2H), 7.46–7.27
(m, 10H), 5.30–5.25 (m, 2H), 4.63 (d, JZ12.4 Hz, 1H), 4.53
(dd, JZ9.8, 9.7 Hz, 1H), 4.06 (d, JZ12.0 Hz, 1H), 3.99 (d,
JZ11.9 Hz, 1H), 3.74 (d, JZ12.4 Hz, 1H), 2.78 (dd, JZ
13.5, 2.7 Hz, 1H), 2.58 (m, 2H), 2.40 (d, JZ13.5, 2.6 Hz,
1H), 2.08 (m, 4H), 1.70 (m, 1H), 1.42–1.26 (m, 10H), 1.10–
0.80 (m, 42H), 0.74 (d, JZ6.7 Hz, 3H), 0.70 (d, JZ6.7 Hz,
3H), 0.37 (s, 6H), 0.32 (s, 6H), 0.06 (s, 6H), K0.65 (s, 6H);
13C NMR (75 MHz, CD2Cl2) d 162.0, 159.6, 147.2, 137.1,
134.3, 134.0, 130.0, 128.2, 126.9, 124.5, 124.1, 115.1, 78.0,
74.0, 66.4, 65.7, 42.6, 35.0, 34.2, 34.0, 33.4, 32.3, 32.1,
29.8, 29.7, 26.9, 26.6, 25.9, 25.8, 25.8, 22.5, 22.5, 18.4,
18.1, 17.7, 17.5, 15.2, 13.9, K4.2, K4.3, K4.3, K4.7,
K5.3; IR (film) 2956, 2929, 2858, 1582, 1455, 1443, 1254,
1136, 1057, 1004, 839, 783, 699; MS (EI) m/z (rel. intensity)
581 ([MC], !1), 566 (2), 526 (16), 525 (43), 524 (100), 506
(9), 494 (13), 398 (10), 386 (5), 342 (3), 332 (3), 284 (4),
256 (2), 75 (2), 73 (19), 67 (2); HR-MS (ESI-pos)
(C34H56NO3Si2) calcd 582.37988, found 582.37987
(MCH).
3.1.5. (R)-6-[2,4-Bis-(tert-butyl-dimethyl-silanyloxy)-5-
phenyl-6-chloro-pyridin-3-yl]-(R)-5-methyl-tetrahydro-
pyran-3-one (25). Prepared analogously. Characteristic
1
data: [a]2D0ZK288 (cZ0.14, CHCl3). H NMR (400 MHz,
CD2Cl2) d 7.47–7.25 (m, 5H), 4.70 (d, JZ10.0 Hz, 1H),
4.15 (dd, JZ15.7, 1.7 Hz, 1H), 3.98 (d, JZ15.7 Hz, 1H),
3.02 (m, 1H), 2.74 (ddd, JZ15.9, 5.1, 1.5 Hz, 1H), 2.20 (dd,
JZ15.9, 11.6 Hz, 1H), 1.01 (s, 9H), 0.89 (s, 9H), 0.83 (m,
3H), 0.38 (s, 3H), 0.38 (s, 3H), 0.02 (s, 3H), K0.57 (s, 3H);
13C NMR (100 MHz, CDCl3) d 207.0, 162.0, 160.2, 147.2,
134.8, 132.4, 128.0, 127.9, 122.8, 113.2, 76.3, 74.3, 46.4,
33.2, 25.9, 25.9, 18.5, 18.1, 17.9, K4.0, K4.1, K4.3, K5.0;
IR (film) 2956, 2931, 2897, 2859, 1733, 1571, 1541, 1428,
1251, 1158, 1140, 1105, 1077, 951, 841, 824, 812, 785, 701;
MS (EI) m/z (rel. intensity) 561 ([MC], !1), 548 (2), 507
(15), 506 (46), 504 (100), 448 (12), 446 (26), 374 (13), 350,
(9), 348 (23), 292 (3), 224 (2), 174 (3), 129 (3), 73 (35); HR-
MS (ESI-pos) (C29H45NO4Si2Cl) calcd 562.257569, found
562.257635 (MCH).
3.1.8. 2,4-Bis-(tert-butyl-dimethyl-silanyloxy)-3-[(R)-3-
methyl-5-(tetradecyliden)-tetrahydro-pyran-2-(R)-yl]-5-
phenyl-pyridine (21b). Prepared analogously using sulfone
20b. 1H NMR (400 MHz, CD2Cl2) d 7.88 (s, 2H), 7.45–7.26
(m, 10H), 5.30–5.25 (m, 2H), 4.62 (d, JZ12.4 Hz, 1H), 4.53
(dd, JZ9.8, 9.7 Hz, 1H), 4.06 (d, JZ12.0 Hz, 1H), 3.98 (d,
JZ11.9 Hz, 1H), 3.75 (d, JZ12.5 Hz, 1H), 2.78 (dd, JZ
13.5, 2.7 Hz, 1H), 2.57 (m, 2H), 2.38, (d, JZ13.5, 2.6 Hz,
1H), 2.02 (m, 5H), 1.68 (m, 1H), 1.43–1.21 (m, 45H), 0.98
(s, 18H), 0.96 (s, 18H), 0.88 (t, JZ6.5 Hz, 6H), 0.72 (d, JZ
6.7 Hz, 3H), 0.69 (d, JZ6.7 Hz, 3H), 0.37 (s, 6H), 0.32 (s,
6H), 0.06 (s, 6H), K0.67 (s, 6H); 13C NMR (100 MHz,
CD2Cl2) d 162.0, 159.6, 147.2, 137.1, 133.9, 130.0, 128.2,
126.9, 124.5, 124.2, 115.1, 78.0, 74.0, 66.4, 65.7, 42.6, 35.0,
34.2, 34.0, 33.4, 32.3, 32.0, 30.2, 29.9, 29.8, 29.8, 29.7,
29.7, 29.5, 29.5, 29.5, 27.2, 27.0, 25.9, 25.8, 25.8, 22.8,
18.4, 18.1, 17.7, 17.5, 14.0, K4.2, K4.3, K4.3, K4.7,
K5.3; IR (film) 2956, 2926, 2855, 1582, 1455, 1444, 1406,
1254, 1136, 1057, 1004, 840, 810, 784, 699; MS (EI) m/z
(rel. intensity) 707 ([MC], !1), 652 (21), 651 (53), 650
(100), 632 (6), 620 (11), 536 (2), 518 (4), 398 (11), 386 (6),
332 (2), 314 (2), 284 (3), 256 (2), 73 (14), 57 (2); HR-MS
3.1.6. 2,4-Bis-(tert-butyl-dimethyl-silanyloxy)-3-[(R)-3-
methyl-5-((S)-6-methyl-octyliden)-tetrahydro-pyran-2-
(R)-yl]-5-phenyl-pyridine (10). A solution of LiHMDS
(100 mg, 0.60 mmol) in DME (2 mL) was added to a
solution of sulfone (S)-19 (160 mg, 0.48 mmol) in DME
(4 mL) at K78 8C. The resulting, bright yellow mixture was
stirred for 15 min before a solution of ketone 9 (200 mg,
0.38 mmol) in DME (4 mL) was introduced and stirring was
continued at that temperature for 30 min. The reaction was
quenched with aq. NH4Cl/NH4OH (pHZ8, 10 mL), the
aqueous layer was repeatedly extracted with EtOAc, the
combined organic layers were dried and evaporated, and
the crude product was purified by flash chromatography
(Et2O/Et3N/pentane, 1:1:100) to give product 10 as a
colorless syrup which is moisture sensitive and immediately
used in the next step (120 mg, 69% over two steps, E/ZZ
1
1:1). Characteristic data: H NMR (300 MHz, CD2Cl2) d
7.89 (s, 2H), 7.45–7.30 (m, 12H), 5.30–5.25 (m, 2H), 4.63
(d, JZ12.4 Hz, 1H), 4.54 (m, 2H), 4.05 (d, JZ12.0 Hz,
1H), 4.00 (d, JZ11.9 Hz, 1H), 3.76 (d, JZ12.5 Hz, 1H),

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A. Furstner et al. / Tetrahedron 60 (2004) 9543–9558
9553
(ESIpos) (C43H74NO3Si2) calcd 708.72053, found
708.71976 (MCH).
5H), 4.34 (d, JZ10.2 Hz, 1H), 3.81 (d, JZ11.3 Hz, 1H),
3.58 (dd, JZ11.3, 2.6 Hz, 1H), 2.60 (m, 1H), 1.80 (d, JZ
10.4 Hz, 1H), 1.63–1.46 (m, 4H), 1.40–1.20 (m, 9H), 1.15–
1.05 (m, 2H), 1.02 (s, 9H), 0.96 (s, 9H), 0.94–0.76 (m, 6H),
0.61 (d, JZ6.7 Hz, 3H), 0.38 (s, 3H), 0.35 (s, 3H), 0.08 (s,
3H), 0.08 (s, 3H); IR (film) 3058, 2955, 2926, 2856, 1646,
3.1.9. 2,4-Bis-(tert-butyl-dimethyl-silanyloxy)-3-[(R)-3-
methyl-5-((S)-6-methyl-octyliden)-tetrahydro-pyran-2-
(R)-yl]-5-phenyl-6-chloro-pyridine. Prepared analogously
1
1582, 1454, 1444, 1256, 1060, 839, 785, 700; MS (EI) m/z
¨
as a E/ZZ1:1 mixture of isomers. Characteristic data: H
C
(rel. Intensitat) 639 ([M ], !1), 624 (3), 584 (19), 583 (48),
NMR (300 MHz, CD2Cl2) d 7.50–7.28 (m, 10H), 7.25 (s, br,
2H), 5.30–5.25 (m, 2H), 4.62 (d, JZ12.5 Hz, 1H), 4.53 (t,
JZ9.7 Hz, 1H), 4.07 (d, JZ12.0 Hz, 1H), 3.98 (d, JZ
11.9 Hz, 1H), 3.76 (d, JZ12.5 Hz, 1H), 2.78 (d, JZ
13.5 Hz, 1H), 2.60 (m, 2H), 2.39 (d, JZ13.5 Hz, 1H), 2.08
(m, 3H), 1.78 (m, 1H), 1.45–1.22 (m, 9H), 1.20–0.79 (m,
58H), 0.74 (d, JZ6.7 Hz, 3H), 0.70 (d, JZ6.7 Hz, 3H), 0.38
(s, 6H), 0.34 (s, 6H), 0.06 (s, 6H), K0.61 (s, 6H); 13C NMR
(75 MHz, CD2Cl2) d 161.8, 160.4, 146.4, 135.1, 134.0,
133.7, 132.4, 131.8, 127.9, 127.7, 124.4, 122.5, 114.5, 77.8,
74.0, 66.4, 42.5, 36.6, 34.9, 34.5, 34.5, 34.2, 33.4, 30.6,
30.3, 29.6, 27.3, 27.1, 27.0, 27.0, 26.1, 25.9, 25.8, 25.8,
22.5, 19.1, 18.5, 18.1, 17.7, 17.5, 11.3, K4.3, K4.4, K4.45,
K4.5, K5.0; IR (film) 2957, 2928, 2857, 1572, 1541, 1427,
1399, 1255, 1158, 1140, 1070, 1004, 953, 841, 825, 812,
784, 700; MS (EI) m/z (rel. intensity) 671 ([MC], !1), 616
(47), 614 (100), 586 (9), 584 (18), 482 (5), 434 (8), 432 (14),
270 (6), 73 (23); HR-MS (ESI-pos) (C38H63NO3Si2Cl) calcd
672.40350, found 672.40365 (MCH).
582 (100), 468 (20), 398 (3), 314 (1), 272 (2), 73 (30), 55
(7); HR-MS (ESI-pos) (C38H66NO3Si2) calcd 640.458126,
found 640.457570 (MCH). For this purpose, TBAF (1 M in
THF, 0.40 mL, 0.38 mmol) was added to a solution of the
crude product in THF (5 mL) and the resulting mixture was
stirred for 5 min. The reaction mixture was adsorbed on
Celite and the product was purified by flash chromatography
(EtOAc/hexanes, 2:1) to give product 11 as a mixture of
diastereomers (52 mg, 69% over two steps, d.r.Z5:1). The
major isomer was separated by preparative HPLC (Nucleo-
sil-5-120-C18/A, : 4.5!125 mm; MeOH/water 80:20,
0.8 mL minK1; 308 K; 8.6 MPa; retention time: 51.64 min).
[a]2D0ZC94.58 (cZ1.0, MeOH). 1H NMR (400 MHz,
CD2Cl2) d 12.77 (s, br, 1H), 9.65 (s, br, 1H), 7.44–7.30
(m, 5H), 7.26 (s, 1H), 4.67 (d, JZ10.5 Hz, 1H), 3.99 (d, JZ
11.5 Hz, 1H), 3.74 (dd, JZ11.6, 2.5 Hz, 1H), 2.10 (m, 1H),
1.80 (d, JZ13.1 Hz, 1H), 1.68–1.47 (m, 4H), 1.40–1.20 (m,
9H), 1.15–1.05 (m, 2H), 0.89–0.80 (m, 9H); 13C NMR
(100 MHz, CD2Cl2) d 164.4, 164.1, 134.2, 133.5, 129.2,
128.2, 127.3, 115.5, 110.1, 81.4, 72.5, 36.6, 36.2, 34.5, 34.2,
31.2, 30.9, 30.1, 29.5, 27.8, 27.1, 19.0, 17.9, 11.2; MS (EI)
m/z (rel. intensity) 411 ([MC], 61), 393 (33), 368 (18), 366
(23), 284 (34), 266 (15), 226 (31), 216 (30), 201 (100), 187
(40), 146 (6), 118 (11), 91 (4), 57 (18), 55 (16), 43 (20), 41
(18); HR-MS (EI) (C26H37NO3) calcd 411.277343, found
411.277067.
3.1.10. 3-((2R,3R)-Tetrahydro-3-methyl-5-(6-methyl-
octyliden)-2H-pyran-2-yl)-4-hydroxy-5-phenyl-6-chloro-
pyridin-2(1H)-one (26). TBAF (1 M in THF, 0.36 ml,
0.36 mmol) was added to a solution of the pyridone described
above (115 mg, 0.171 mmol) in THF (5 mL) and the resulting
mixture was stirred for 30 min. The solution was adsorbed on
Celite and the product was purified by flash chromatography
(hexanes/EtOAc, 5:1) to afford product 26 as a colorless solid
(74 mg, 99%, E/ZZ1:1). Characteristic data: 1H NMR
(300 MHz, CDCl3) d 10.5 (br, 2H), 7.50–7.27 (m, 10H),
5.28 (m, 2H), 4.85 (m, 2H), 4.71 (d, JZ12.5 Hz, 1H), 4.12 (m,
2H), 3.91 (d, JZ12.7 Hz, 1H), 3.00 (m, 1H), 2.72 (d, JZ
11.5 Hz, 1H), 2.35 (dd, JZ13.0, 2.1 Hz, 2H), 2.20–1.75 (m,
10H), 1.40–0.70 (m, 34H); 13C NMR (75 MHz, CDCl3) d
164.8, 162.8, 138.3, 132.6, 132.3, 132.1, 130.7, 128.1, 127.9,
126.0, 116.0, 107.0, 80.2, 67.1, 41.6, 38.3, 37.6, 36.4, 34.3,
34.3, 34.2, 30.2, 29.8, 29.5, 29.4, 27.1, 26.8, 26.6, 19.1, 18.0,
17.8, 11.4, 11.3; IR(film)3200, 2959, 2926, 2854, 1635, 1614,
1449, 1359, 1236, 1050, 954, 838, 762, 697; MS (EI) m/z (rel.
intensity) 443 ([MC], 57), 427 (9), 425 (23), 408 (5), 387 (17),
358 (9), 330 (12), 326 (16), 261 (51), 250 (100), 237 (12), 234
(43), 221 (22), 144 (8), 82 (14), 55 (21); HR-MS (ESI-pos)
(C26H34NO3ClNa) calcd 466.21249, found 466.21292
(MCNa).
3.1.12. 3-((2R,3R,5R)-Tetrahydro-3-methyl-5-pentyl-
2H-pyran-2-yl)-4-hydroxy-5-phenyl-pyridin-2(1H)-one
(22a, nZ1). Prepared analogously. The major isomer was
purified by preparative HPLC (Nucleosil-100-5-C18/A, :
4.5!125 mm; MeOH/water 75:25; 0.8 mL minK1; 308 K;
12.1 MPa). [a]2D0Z98.58 (cZ0.51, CHCl3). 1H NMR
(400 MHz, CD2Cl2) d 12.65 (s, br, 1H), 9.70 (s, br, 1H),
7.48–7.26 (m, 5H), 7.25 (s, 1H), 4.66 (d, JZ10.5 Hz, 1H),
3.99 (d, JZ11.5 Hz, 1H), 3.74 (dd, JZ11.6, 2.4 Hz, 1H),
2.10 (m, 1H), 1.78 (d, JZ11.7 Hz, 1H), 1.68–1.40 (m, 3H),
1.40–1.20 (m, 7H), 0.95–0.80 (m, 6H); 13C NMR
(100 MHz, CD2Cl2) d 164.3, 164.1, 134.2, 133.5, 129.2,
128.2, 127.3, 115.5, 110.1, 81.4, 72.5, 36.2, 34.2, 32.0, 31.2,
30.8, 27.5, 22.7, 17.9, 14.0; IR (film) 3131, 2956, 2926,
2855, 1646, 1600, 1457, 1233, 1053, 698; MS (EI) m/z (rel.
intensity) 355 ([MC], 43), 337 (24), 325 (7), 312 (18), 310
(17), 284 (24), 266 (17), 242 (22), 229 (12), 226 (32), 216
(29), 201 (100), 200 (76), 187 (44), 155 (10), 146 (6), 144
(5), 130 (6), 118 (10), 91 (7), 55 (14), 41 (13); HR-MS (EI)
(C22H29NO3) calcd 355.214744, found 355.214445.
3.1.11. 3-((2R,3R,5R)-Tetrahydro-3-methyl-5-((S)-6-
methyloctyl)-2H-pyran-2-yl)-4-hydroxy-5-phenylpyri-
din-2(1H)-one (11). A suspension of compound 10 (E/Z
mixture, 120 mg, 0.19 mmol) and Pd/C (10% w/w, 30 mg)
in EtOH (6 mL) was stirred under an atmosphere of H2
(1 atm) for 24 h. The catalyst was filtered off and was
carefully washed with hot EtOH (containing 1% Et3N) and
EtOAc (containing 1% Et3N), the combined filtrates were
evaporated and the crude product was directly subjected to
the subsequent desilylation reaction. Characteristic data: 1H
NMR (400 MHz, CD2Cl2) d 7.86 (s, 1H), 7.43–7.28 (m,
3.1.13. 3-((2R,3R,5R)-Tetrahydro-3-methyl-5-tetradecyl-
2H-pyran-2-yl)-4-hydroxy-5-phenylpyridin-2(1H)-one
(22b, nZ10). Prepared analogously. The major isomer was
purified by preparative HPLC (Nucleosil-100-5-C18/A :
4.5!125 mm; MeOH/water 95:5; 0.8 mL minK1; 308 K;
6.1 MPa; retention time 14.271 min). [a]2D0Z68.68 (cZ0.55,
CHCl3). 1H NMR (400 MHz, CDCl3) 9.82 (s, br, 1H),

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A. Furstner et al. / Tetrahedron 60 (2004) 9543–9558
9554
7.48–7.26 (m, 6H), 4.68 (d, JZ10.5 Hz, 1H), 3.99 (d, JZ
11.5 Hz, 1H), 3.74 (dd, JZ11.6, 2.3 Hz, 1H), 2.10 (m, 1H),
1.79 (d, JZ12.3 Hz, 1H), 1.68–1.44 (m, 4H), 1.40–1.18 (m,
24H), 0.95–0.80 (m, 6H); 13C NMR (100 MHz, CDCl3) d
164.3, 163.2, 133.6, 133.4, 129.1, 128.4, 127.7, 116.9,
109.6, 81.2, 72.5, 36.1, 34.0, 31.9, 31.0, 30.9, 29.7 (m), 29.6,
29.6, 29.3, 27.8, 22.7, 18.0, 14.1; IR (film) 3143, 2958,
2922, 2852, 1645, 1601, 1457, 1435, 1380, 1230, 1051, 801,
759, 697; MS (EI) m/z (rel. intensity) 481 ([MC], 21), 463
(23), 451 (6), 438 (13), 436 (14), 382 (2), 298 (9), 284 (20),
266 (11), 242 (23), 229 (10), 228 (17), 227 (9), 226 (22), 216
(21), 201 (100), 200 (64), 187 (20), 146 (3), 118 (4), 97 (3),
95 (2), 91 (3), 67 (2), 43 (14); HR-MS (ESI-pos)
(C31H48NO3) calcd 482.363419, found 482.36352 (MCH).
27.1, 19.2, 17.8, 11.4; IR (film) 3180, 2958, 2925, 2854,
1640, 1580, 1556, 1455, 1381, 1220, 1054, 775, 698, 658;
MS (EI) m/z (rel. intensity) 427 ([MC], 32), 410 (72), 392
(60), 382 (14), 380 (11), 368 (10), 314 (5), 284 (12), 282 (4),
266 (11), 258 (10), 244 (11), 242 (16), 240 (15), 238 (11),
232 (11), 226 (27), 216 (59), 214 (34), 211 (13), 200 (100),
187 (19), 144 (4), 97 (5), 67 (4), 55 (15), 43 (14), 41 (13);
HR-MS (EI) (C26H37NO4) calcd 427.272259, found
427.272083.
3.1.16. 3-((2R,3R,5R)-Tetrahydro-3-methyl-5-((R)-6-
methyloctyl)-2H-pyran-2-yl)-1,4-dihydroxy-5-phenyl-
pyridin-2(1H)-one, (17R)-TMC-69-6H, ((17R)-2). Pre-
pared analogously. [a]2D0Z102.38 (cZ0.6, MeOH). 1H
NMR (600 MHz, CDCl3) d 9.52 (s, br, 1H), 7.67 (s, 1H),
7.47–7.31 (m, 5H), 4.68 (d, JZ10.5 Hz, 1H), 3.96 (d, JZ
11.8 Hz, 1H), 3.72 (dd, JZ11.6, 2.5 Hz, 1H), 2.07 (m, br,
1H), 1.76 (d, JZ11.0 Hz, 1H), 1.63 (m, br, 1H), 1.58–1.44
(m, 3H), 1.35–1.18 (m, 9H), 1.13–1.02 (m, 2H), 0.87–0.81
(m, 6H), 0.79 (d, JZ6.6 Hz, 3H); 13C NMR (150 MHz,
CDCl3) d 161.0, 157.2, 133.1, 130.5, 129.2, 128.4, 127.7,
113.7, 109.5, 81.6, 72.4, 36.5, 36.1, 34.4, 34.0, 31.0, 30.8,
29.9, 29.5, 27.8, 27.0, 19.2, 17.8, 11.4; IR (film) 3187, 2958,
2924, 2854, 1643, 1580, 1557, 1456, 1381, 1220, 1054, 984,
775, 699; MS (EI) m/z (rel. intensity) 427 ([MC], 45), 410
(100), 392 (74), 382 (15), 380 (12), 367 (9), 314 (6), 284 (5),
282 (5), 266 (7), 258 (13), 244 (13), 242 (10), 240 (17), 238
(11), 232 (16), 226 (18), 216 (66), 214 (37), 211 (14), 200
(87), 187 (12), 144 (5), 97 (6), 57 (15), 55 (22), 43 (22), 41
(21); HR-MS (EI) (C26H37NO4) calcd 427.272259, found
427.271906.
3.1.14. 3-((2R,3R,5R)-Tetrahydro-3-methyl-5-(6-methyl-
octyl)-2H-pyran-2-yl)-4-hydroxy-5-phenyl-6-chloro-
pyridin-2(1H)-one (27). Prepared analogously. The major
isomer was purified by preparative HPLC (Asahipak, :
4.5!250 mm; MeCN/TEAA buffer (pH 6.96)Z70:30;
0.8 mL minK1; 308 K; 6.5 MPa). [a]2D0ZK83.08 (cZ0.7,
CHCl3). 1H NMR (400 MHz, CD2Cl2) d 7.50–7.29 (m, 5H),
4.64 (d, JZ10.5 Hz, 1H), 3.94 (d, JZ11.8 Hz, 1H), 3.72
(dd, JZ11.8, 2.6 Hz, 1H), 2.04 (m, 1H), 1.77 (d, JZ
12.7 Hz, 1H), 1.70–1.38 (m, 4H), 1.35–1.17 (m, 10H), 1.19–
1.00 (m, 2H), 0.93–0.78 (m, 9H); 13C NMR (100 MHz,
CDCl3) d 165.0, 162.7, 139.2, 132.9, 130.8, 128.2, 128.1,
128.0, 116.4, 107.5, 81.0, 72.4, 36.6, 36.2, 34.4, 34.0, 31.1,
30.8, 29.4, 27.8, 27.0, 19.2, 18.0, 11.4; IR (film) 3189, 2959,
2926, 2855, 1634, 1614, 1590, 1460, 1380, 1263, 1235,
1076, 1056, 982, 762, 698; MS (EI) m/z (rel. intensity) 445
([MC], 100), 427 (17), 402 (14), 400 (14), 318 (16), 276
(19), 275 (10), 274 (11), 223 (17), 221 (42), 211 (19), 144
(11), 129 (5), 116 (6), 115 (6), 82 (7), 57 (19), 43 (28), 41
(29); HR-MS (ESI-pos) (C26H37NO3Cl) calcd 446.24619,
found 446.24617.
3.1.17. 3-((2R,3R,5R)-Tetrahydro-3-methyl-5-(pentyl)-
2H-pyran-2-yl)-1,4-dihydroxy-5-phenyl-pyridin-2(1H)-
one (23a, nZ1). Prepared analogously. [a]2D0Z88.88 (cZ
1
0.8, CH2Cl2). H NMR (300 MHz, CDCl3) d 9.52 (s, br,
1H), 7.67 (s, 1H), 7.50–7.30 (m, 5H), 4.68 (d, JZ10.5 Hz,
1H), 3.96 (d, JZ11.5 Hz, 1H), 3.73 (dd, JZ11.6, 2.5 Hz,
1H), 2.08 (br, m, 1H), 1.76 (d, JZ11.0 Hz, 1H), 1.68–1.42
(m, 4H), 1.38–1.19 (m, 6H), 0.92–0.86 (t, JZ7.0 Hz, 3H),
0.82 (d, JZ6.2 Hz, 3H); 13C NMR (75 MHz, CDCl3) d
160.9, 157.1, 133.1, 130.3, 129.2, 128.4, 127.7, 113.7,
109.5, 81.6, 72.4, 36.1, 34.0, 31.0, 30.8, 29.7, 27.6, 22.4,
17.9, 14.0; IR (film) 3181, 2957, 2922, 2852, 1736, 1639,
1579, 1556, 1455, 1381, 1260, 1218, 1048, 801, 775, 697;
MS (EI) m/z (rel. intensity) 371 ([MC], 24), 354 (39), 336
(29), 312 (13), 284 (17), 266 (15), 242 (17), 216 (43), 200
(100), 187 (30), 155 (13), 118 (9), 97 (10), 69 (12), 55 (22);
HR-MS (EI) (C22H29NO4) calcd 371.209659, found
371.209745.
3.1.15. 3-((2R,3R,5R)-Tetrahydro-3-methyl-5-((S)-6-
methyloctyl)-2H-pyran-2-yl)-1,4-dihydroxy-5-phenyl-
pyridin-2(1H)-one, (17S)-TMC-69-6H, ((17S)-2). A
mixture of pyridone 11 (25 mg, 0.06 mmol), HMDS
(0.50 mL) and freshly distilled TMSCl (9 mL) was refluxed
for 6 h before all volatile components were removed in
vacuo. The remaining yellow syrup was dissolved in
CH2Cl2 (1 mL) and treated with (pyridine)MoO5(HMPA)
(52.5 mg, 0.12 mmol).34 After the mixture had been stirred
for 16 h, the mixture was diluted with EtOAc (2.5 mL) and
sat. aq. EDTA-Na4 (2.5 mL), causing a color change from
dark red to pale yellow. The pH of the solution was adjusted
to 7–7.5 upon addition of aq. HCl (0.1 M) before the
aqueous phase was separated and repeatedly extracted with
EtOAc. The combined organic layers were evaporated and
the residue was dried in vacuo (10K4 Torr) to provide
compound (17S)-2 as a colorless powder (18.9 mg, 71%).
[a]2D0Z88.68 (cZ0.95, MeOH). 1H NMR (400 MHz,
CDCl3) d 9.52 (s, br, 1H), 7.67 (s, 1H), 7.47–7.29 (m,
5H), 4.68 (d, JZ10.5 Hz, 1H), 3.96 (d, JZ11.5 Hz, 1H),
3.72 (dd, JZ11.5, 2.3 Hz, 1H), 2.06 (m, br, 1H), 1.76 (d,
JZ12.2 Hz, 1H), 1.62 (m, br, 1H), 1.58–1.40 (m, 3H), 1.35–
1.19 (m, 9H), 1.13–1.00 (m, 2H), 0.89–0.81 (m, 6H), 0.79
(d, JZ6.6 Hz, 3H); 13C NMR (100 MHz, CDCl3) d 160.9,
157.2, 133.2, 131.1, 129.2, 128.4, 127.7, 113.5, 109.3, 81.6,
72.4, 36.5, 36.1, 34.4, 33.9, 31.0, 30.8, 30.0, 29.5, 27.8,
3.1.18. 3-((2R,3R,5R)-Tetrahydro-3-methyl-5-(tetra-
decyl)-2H-pyran-2-yl)-1,4-dihydroxy-5-phenylpyridin-
2(1H)-one (23b, nZ10). Prepared analogously. [a]2D0Z
66.08 (cZ1.9, CH2Cl2). 1H NMR (300 MHz, CDCl3) d 9.63
(s, br, 1H), 7.67 (s, 1H), 7.48–7.27 (m, 5H), 4.69 (d, JZ
10.5 Hz, 1H), 3.98 (d, JZ11.4 Hz, 1H), 3.73 (d, JZ11.7,
1H), 2.10 (m, 1H), 1.78 (d, JZ12.5 Hz, 1H), 1.69–1.42 (m,
4H), 1.37–1.18 (m, 24H), 0.92–0.80 (m, 6H); IR (film)
3136, 2923, 2853, 1644, 1601, 1581, 1555, 1456, 1381,
1221, 1053, 801, 698; MS (ESI) m/z (rel. intensity) 497
([MC], !1), 482 (100), 433 (6), 390 (4), 180 (13); HR-MS

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A. Furstner et al. / Tetrahedron 60 (2004) 9543–9558
9555
(ESI-pos) (C31H48NO4) calcd 498.35833, found 498.35885
(MCH).
chromatography (hexanes/Et2O, 10:1) afforded product 16
as a colorless liquid (650 mg, 87%). [a]2D0ZC16.28 (cZ
1.0, CHCl3/MeOHZ9:1). H NMR (400 MHz, CDCl3) d
1
6.97 (dt, JZ15.6, 7.0 Hz, 1H), 5.81 (m, 1H), 5.70–5.60 (m,
1H), 5.01–4.90 (m, 2H), 3.72 (s, 3H), 2.18–2.05 (m, 3H),
1.47–1.40 (m, 2H), 1.01 (d, JZ6.7 Hz, 3H); 13C NMR
(400 MHz, CDCl3) d 166.8, 149.2, 143.4, 120.6, 113.0,
51.0, 37.0, 34.4, 29.6, 19.8; IR (film) 3077, 2954, 2928,
1728, 1658, 1436, 1273, 994, 913; MS (EI) m/z (rel.
intensity) 168 ([MC], !1), 153 (17), 139 (16), 126 (8), 113
(53), 111 (23), 109 (35), 100 (84), 94 (50), 87 (19), 81 (43),
79 (27), 71 (13), 69 (65), 59 (21), 55 (78), 41 (100), 29 (23);
HR-MS (ESI-pos) (C10H16O2) calcd 169.122855, found
169.122843 (MCH).
3.1.19. 3-((2R,3R,5R)-Tetrahydro-3-methyl-5-(6-methyl-
octyl)-2H-pyran-2-yl)-1,4-dihydroxy-5-phenyl-6-chloro-
pyridin-2(1H)-one (28). Prepared analogously. [a]2D0ZC
24.08 (cZ0.5, CHCl3). 1H NMR (600 MHz, CDCl3) d 9.96
(s, br, 1H), 9.38 (s, br, 1H), 7.50–7.25 (m, 5H), 4.62 (d, JZ
10.2 Hz, 1H), 3.91 (d, JZ11.0 Hz, 1H), 3.67 (d, JZ
11.0 Hz, 1H), 2.02 (m, br, 1H), 1.73 (d, JZ12.1 Hz, 1H),
1.62–1.37 (m, 4H), 1.36–1.00 (m, 11H), 0.86–0.70 (m, 9H);
13C NMR (150 MHz, CDCl3) d 160.7, 156.8, 130.8, 128.4,
128.3, 128.2, 113.0, 107.0, 81.5, 72.4, 36.5, 36.1, 34.4, 33.9,
31.1, 30.8, 30.2, 29.9, 29.5, 27.8, 27.0, 19.2, 17.8, 11.4; IR
(film) 3168, 2959, 2924, 2854, 1625, 1532, 1453, 1379,
1260, 1215, 1090, 1054, 912, 802, 698; MS (EI) m/z (rel.
intensity) 461 ([MC], 16), 446 (35), 445 (48), 444 (73), 428
(12), 427 (13), 426 (26), 416 (11), 400 (6), 398 (5), 320 (2),
318 (6), 302 (4), 300 (8), 276 (13), 274 (11), 260 (12), 252
(12), 250 (44), 234 (100), 221 (20), 211 (13), 180 (4), 144
(7), 55 (14), 43 (12), 41 (11); HR-MS (ESI-pos)
(C26H37NO4Cl) calcd 462.24111, found 462.24080
(MCH).
3.1.23. (R)-6-Methyloctan-1-ol (17). Dibal-H (1.0 M in
hexane, 25.6 mL, 25.6 mmol) was added dropwise to a
solution of ester 16 (1.50 g, 8.92 mmol) in Et2O (20 mL) at
0 8C. After stirring for 2 h, the reaction was carefully
quenched with MeOH at K788 C. A standard extractive
work up followed by flash chromatography (Et2O/hexanes,
1:2) afforded (S,E)-6-methylocta-2,7-dien-1-ol as a color-
less liquid (1.13 g, 91%) which showed the following
analytical and spectroscopic properties: [a]2D0ZC14.08
(cZ1.0, CHCl3/MeOHZ9:1). 1H NMR (400 MHz,
CDCl3) d 5.72–5.58 (m, 3H), 5.01–4.90 (m, 2H), 4.10–
4.05 (m, 2H), 2.18–1.97 (m, 3H), 1.43–1.33 (m, 2H), 0.99
(d, JZ6.7 Hz, 3H); 13C NMR (100 MHz, CDCl3) d 144.4,
133.3, 129.0, 112.8, 63.8, 37.2, 35.9, 29.8, 20.1; IR (film)
3329, 3077, 2961, 2925, 1640, 1454, 1373, 971; MS (EI)
m/z (rel. intensity) 140 ([MC], !1), 122 (3), 109 (17), 107
(22), 93 (37), 83 (28), 79 (25), 70 (22), 68 (64), 67 (70), 57
(30), 55 (97), 51 (5), 41 (100), 31 (11), 29 (41); HR-MS (CI)
(C9H16O) calcd 141.127940, found 141.127788 (MCH). A
suspension of this alcohol (100 mg, 0.70 mmol) and Pd/C
(10% w/w, 20 mg) in MeOH (5 mL) was stirred for 6 h
under an atmosphere of H2 (1 atm). The catalyst was filtered
off and was carefully rinsed with CH2Cl2, the combined
filtrates were evaporated and the crude product was purified
by flash chromatography (Et2O/hexanes, 2:1) to give
3.1.20. 3-Dodecyl-1,4-dihydroxy-5-phenyl-1H-pyridin-2-
one (32). Prepared analogously from pyridone 31. 1H NMR
(300 MHz, CDCl3) d 7.60–7.29 (m, 6H), 2.68 (m, 2H),
2.10–1.50 (m, 2H), 1.40–1.15 (m, 18H) 0.88 (t, JZ6.9 Hz,
3H); 13C NMR (75 MHz, CDCl3) d (signals for C-2 and C-4
were not detected), 132.3, 129.8, 128.7, 128.2, 127.8, 118.0,
113.9, 42.0, 31.9, 29.7, 29.6, 29.6, 29.4, 29.3, 29.3, 22.7,
22.7, 14.1; IR (film) 3252, 2921, 2852, 1716, 1622, 1543,
1497, 1463, 1344, 1262, 1198, 1088, 771, 697; MS (EI) m/z
(rel. intensity) 371 ([MC], 33), 354 (3), 272 (2), 216 (7), 200
(6), 176 (12), 175 (100), 146 (20), 118 (10), 91 (5), 55 (6),
43 (9); HR-MS (ESI-pos) (C23H33NO3Na) calcd 394.23581,
found 394.23593 (MCNa).
3.1.21. (S)-4-Methylhex-5-enal (15).29 Ozone was bubbled
through a solution of (C)-(S)-citronellene 14 (8.2 g,
59 mmol, 91% ee) in CH2Cl2 (120 mL) at K78 8C until
TLC showed complete consumption of the starting material.
At this point, Me2S (7.0 mL, 148 mmol) was added and the
resulting mixture was stirred for 1 h at K30 8C. Excess
Me2S was pumped off in vacuo at that temperature before all
volatile components were evaporated at ambient tempera-
ture and the crude product was purified by distillation (bp
52–55 8C, 50 mbar) to give aldehyde 15 as a colorless liquid
1
product 17 as a colorless liquid (90 mg, 88%). H NMR
(400 MHz) (CDCl3) d 3.59 (m, 2H), 1.56–1.51 (m, 3H),
1.37–1.25 (m, 4H), 1.20–1.07 (m, 4H), 0.92–0.86 (m, 6H);
13C NMR (100 MHz) (CDCl3) d 63.0, 36.7, 34.5, 33.1, 29.6,
27.0, 26.2, 19.1, 11.3.
3.1.24. 5-((R)-6-Methyloctylthio)-1-phenyl-1H-tetrazol
((R)-18).41 DEAD (202 mg, 1.00 mmol) was added to a
solution of alcohol 17 (80 mg, 0.55 mmol), 1-phenyl-1H-
tetrazole-5-thiol (45) (198 mg, 1.10 mmol) and PPh3
(218 mg, 0.83 mmol) in THF (5 mL) and the resulting
mixture was stirred for 10 min. A standard extractive work
up followed by flash chromatography (hexanes/EtOAcZ
30:1) afforded sulfide 18 as a colorless syrup (114 mg, 68%)
which was immediately subjected to the subsequent
oxidation. Characteristic data: [a]2D0ZK2.58 (cZ1.0,
CHCl3). 1H NMR (400 MHz, CDCl3) d 7.57–7.53 (m,
5H), 3.38 (m, 2H), 1.83–1.78 (m, 2H), 1.44–1.06 (m, 9H),
0.86–0.82 (m, 6H); 13C NMR (100 MHz, CDCl3) d 154.4,
133.8, 130.0, 129.7, 123.8, 36.3, 34.2, 33.4, 29.4, 29.1, 28.9,
26.5, 19.1, 11.3; IR (film) 2958, 2927, 2856, 1597, 1500,
1462, 1386, 1243, 761, 694, 552.
1
(6.45 g, 98%). [a]D20ZC3.58 (cZ1.0, CHCl3). H NMR
(400 MHz, CDCl3) d 9.78 (s, 1H), 5.70–5.58 (m, 1H), 5.02–
4.94 (m, 2H), 2.46–2.40 (m, 2H), 2.20–2.10 (m, 1H), 1.72–
1.53 (m, 2H), 1.02 (d, JZ6.7 Hz, 3H); 13C NMR (100 MHz,
CDCl3) d 202.6, 143.3, 113.8, 41.8, 37.4, 28.5, 20.2.
3.1.22. (S,E)-Methyl 6-methylocta-2,7-dienoat (16).
Methyl diethylphosphonoacetate (1.41 g, 6.69 mmol) was
added dropwise to a suspension of NaH (160 mg,
6.69 mmol) in THF (15 mL) at 0 8C and the resulting
mixture was stirred for 30 min at ambient temperature. The
solution was cooled to K78 8C before aldehyde 15 (500 mg,
4.46 mmol) was introduced via syringe and the resulting
mixture was allowed to reach ambient temperature. A
standard extractive work up followed by flash

¨
A. Furstner et al. / Tetrahedron 60 (2004) 9543–9558
9556
3.1.25. 5-((R)-6-Methyloctylsulfonyl)-1-phenyl-1H-tetra-
zol ((R)-19). A solution of (NH4)6Mo7O24$4H2O (550 mg,
0.45 mmol) in aq. H2O2 (30%, 5.84 g, 51.50 mmol) was
added to a solution of sulfide (R)-18 (1.60 g, 5.26 mmol) in
EtOH (25 mL) and the resulting mixture was stirred for 18 h
at ambient temperature. For work up, the mixture was
diluted with water and EtOAc, the organic phase was
washed with aqueous sodium thiosulfate (5%, w/w) before it
was dried over MgSO4 and evaporated. The residue was
purified by flash chromatography (EtOAc/hexanes, 1:8) to
give sulfone (R)-19 as a colorless syrup (1.64 g, 93%).
[a]2D0ZK2.78 (cZ3.0, CHCl3). 1H NMR (400 MHz,
CDCl3) d 7.72–7.55 (m, 5H), 3.73 (m, 2H), 1.96 (m, 2H),
1.56–1.42 (m, 2H), 1.41–1.23 (m, 5H), 1.20–1.06 (m, 2H),
0.90–0.83 (m, 6H); 13C NMR (100 MHz, CDCl3) d 153.3,
133.0, 131.4, 129.7, 125.1, 56.1, 36.1, 34.3, 29.4, 28.5, 26.4,
22.0, 19.1, 11.3; IR (film) 2958, 2928, 2872, 2859, 1498,
1463, 1341, 1152, 763, 688, 629; MS (EI) m/z (rel. intensity)
336 ([MC], !1), 243 (2), 173 (9), 160 (3), 147 (12), 145
(10), 119 (17), 118 (100), 117 (28), 97 (6), 91 (7), 77 (9), 70
(6), 65 (11), 57 (25), 55 (23), 43 (33), 41 (23), 29 (11);
HR-MS (CI) (C16H24N4SO2) calcd 337.169409, found
337.169423 (MCH); Anal. calcd for C16H23N4SO2: C
57.12, H 7.19, N 16.65, found: C 57.19, H 7.11, N
16.73.
under an atmosphere of H2 (1 atm) for 72 h. The catalyst
was filtered off through a short pad of Celite and was
successively washed with hot MeOH and hot EtOAc, the
combined filtrates were evaporated and the crude product
was purified by HPLC (Nucleosil-100-5-C18/A, : 4.5!
125 mm; MeOH/water 90:10; 0.8 mL minK1; 308 K;
12.5 MPa; retention time: 4.88 min) to give compound 31
1
as a colorless solid (60%). H NMR (400 MHz, CD2Cl2C
CD3OD) d 7.45–7.32 (m, 6H), 2.55 (m, 2H), 1.51–1.40 (m,
2H), 1.42–1.17 (m, 18H), 0.83 (t, JZ6.4 Hz, 3H); 13C NMR
(100 MHz, CD2Cl2CCD3OD) d 166.1, 162.8, 135.5, 132.1,
130.5, 129.8, 128.9, 117.3, 114.6, 33.1, 31.0, 30.9, 30.8,
30.6, 29.6 (m), 24.5, 23.9, 14.9; IR (film) 2923, 2852, 1640,
1620, 1600, 1455, 1434, 1208, 1132, 1082, 881, 760, 699;
MS (EI) m/z (rel. intensity) 355 ([MC], 17), 338 (10), 214
(13), 201 (100), 200 (46), 188 (1), 173 (1), 146 (2), 130 (3),
118 (2), 91 (3), 55 (3), 43 (3), 41 (2); HR-MS
(C23H33NO2CNa) calcd 378.24054, found 378.24059
(MCNa).
3.2. Enzymatic assays
PTP1B-inhibition. PTP1B was purchased from Calbiochem
(human recombinant). The enzyme (0.001 U) was pre-
incubated with the inhibitors in a buffer (pH 7.2)42
containing HEPES (25 mM), EDTA (2.5 mM), NaCl
(50 mM), DTT (2 mM) and BSA (0.1%) for 15 min at
room temperature. Then p-NPP was added (end concen-
tration 50 mM) and the read-out (405 nm) was recorded on a
microplate-reader at 37 8C continuously for 80 min. The
reaction rate was determined from the absorption difference
between 30 and 60 min reaction time.
3.1.26. 3-Allyl-4-hydroxy-5-phenylpyridin-2(1H)-one
(30a). A solution of allyl acetate 29a (59 mg, 0.59 mmol)
in DMF (2 mL) was slowly added to a solution of pyridone 5
(100 mg, 0.53 mmol) and Pd(PPh3)4 (31 mg, 0.03 mmol) in
DMF (3 mL) and Et3N (0.91 ml, 0.57 mmol), and the
resulting mixture was stirred at 110 8C for 2 h. A standard
extractive work up followed by flash chromatography of the
crude product (EtOAc/EtOAc/MeOH, 20:1) provided
Cdc25A-inhibition. The clone pET9d/His-Cdc25A was
expressed in the E. coli strain BL21-DE3 and purified in
the presence of 8M urea. 30 mg of the purified enzyme was
pre-incubated with the inhibitors in a buffer pH 8.0
containing 50 mM Tris, 50 mM NaCl and 2 mM DTE for
15 min at room temperature.43,44
1
product 30a as a colorless solid (71 mg, 59%). H NMR
(400 MHz, DMSO-d6) d 11.28 (s, br, 1H), 9.45 (s, br, 1H),
7.68–7.24 (m, 6H), 5.82 (m, 1H), 5.01 (dd, JZ2.1, 17.2 Hz,
1H), 4.92 (dd, JZ2.1, 10.1 Hz, 1H), 3.30 (d, JZ6.0 Hz,
2H); 13C NMR (75 MHz, CDCl3) d 163.1, 160.4, 136.1,
135.3, 132.0, 129.2, 128.3, 126.8, 114.4, 113.9, 109.4, 27.1;
IR (film) 3051, 1643, 1479, 1433, 1270, 1095, 756, 728,
694; MS (EI) m/z (rel. intensity) 227 ([MC], 61), 212 (100),
200 (5), 144 (3), 128 (4), 118 (14), 106 (2), 63 (4), 51 (4);
HR-MS (EI) (C14H13NO2) calcd 227.094629, found
227.094368.
3.3. X-ray crystallographic study
Suitable crystals were obtained by recrystallization from
n-hexane (8) and n-heptane (25). Data were recorded using
an Enraf-Nonius KappaCCD diffractometer with graphite-
˚
monochromated Mo Ka-radiation (lZ0.71073 A). The
3.1.27. 3-(Dodec-2-enyl)-4-hydroxy-5-phenylpyridin-
2(1H)-one (30b). Prepared analogously as a mixture of
isomers (E/ZZ1:1). H NMR (400 MHz, CDCl3) d 7.80–
crystal was mounted in a stream of cold nitrogen gas. The
structures were solved by direct methods (SHELXS-97)45
and refined by full-matrix least-squares techniques against
F2 (SHELXL-97).46 Hydrogen atoms were inserted from
geometry consideration using the HFIX option of the
program. Crystallographic data (excluding structure factors)
have been deposited with the Cambridge Crystallographic
Data Centre as supplementary publication number CCDC
236780 (8) and 236781 (25). Copies of the data can be
obtained, free of charge, on application to CCDC, 12 Union
Road, Cambridge CB2 1 EZ, UK (fax: C44-1223-336033
or e-mail: deposit@ccdc.cam.ac.uk).
1
7.20 (m, 12H), 5.62 (m, 4H), 3.43 (d, JZ6.1, 4H), 2.00 (m,
4H), 1.47–1.13 (m, 24H), 0.87 (t, JZ7.0 Hz, 6H); 13C NMR
(100 MHz, CDCl3) d 164.5, 162.0, 132.1, 132.0, 129.1,
128.9, 128.8, 128.5, 128.4, 127.8, 126.3, 115.7, 110.2, 31.4,
31.8, 29.5, 29.4, 29.3, 29.2 (m), 27.3, 26.4, 22.6, 14.0; MS
(EI) m/z (rel. intensity) 353 ([MC], 35), 336 (4), 324 (3), 282
(4), 241 (11), 240 (35), 226 (25), 224 (5), 212 (29), 200
(100), 187 (7), 146 (4), 118 (6), 91 (7), 77 (3), 55 (6), 43
(10); HR-MS (EI) (C23H31NO2): calcd 353.235479, found
353.235538.
Selected X-ray crystallographic data for compound 8.
3.1.28. 3-Dodecyl-4-hydroxy-5-phenyl-1H-pyridin-2-one
(31). A suspension of olefin 30b (83 mg, 0.23 mmol) and
Pd/C (20 mg, 10%, w/w) in MeOH (2.5 mL) was stirred
C28H41NO4Si2, MrZ511.80 g molK1 colorless, crystal
size 0.44!0.08!0.06 mm, orthorhombic, P21212 [No.
18], aZ18.2689(2), bZ26.2118(3), cZ12.40800(10) A,
,
˚

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A. Furstner et al. / Tetrahedron 60 (2004) 9543–9558
9557
3
VZ5941.70(11) A , ZZ8, DcalcZ1.144 Mg mK3, mZ
0.150 mmK1, TZ100 K, 74260 reflections collected,
13582 independent reflections, 8780 reflections with IO
2s(I), qmaxZ27.458, 651 refined parameters, RZ0.0686,
RwZ0.1666, SZ1.492, largest diff. peak and holeZ
˚
240, 236–243. (c) Gasparotto, D.; Maestro, R.; Piccinin, S.;
Vukosavljevic, T.; Barzan, L.; Sulfaro, S.; Boiocchi, M.
Cancer Res. 1997, 57, 2366–2368. (d) Wu, W.; Fan,
Y. H.; Kemp, B. L.; Walsh, G.; Mao, L. Cancer Res.
1998, 58, 4082–4085.
K3
˚
10. (a) Hirano, N.; Kohno, J.; Tsunoda, S.; Nishio, M.; Kishi, N.;
Okuda, T.; Kawano, K.; Komatsubara, S.; Nakanishi, N.
J. Antibiot. 2001, 54, 421–427. (b) Kohno, J.; Hirano, N.;
Sugawara, K.; Nishio, M.; Hashiyama, T.; Nakanishi, N.;
Komatsubara, S. Tetrahedron 2001, 57, 1731–1735.
(c) Sugawara, K.; Kohno, J.; Nakanishi, N.; Hashiyama, T.
Tennen Yuki Kagobutsu Toronkai Koen Yoshishu 2000, 42,
661–666. Chem. Abstr. 2001, 134, 340370.
0.547/K0.503 e A
.
Selected X-ray crystallographic data for compound 25.
C29H44ClNO4Si2, MrZ562.28 g molK1, colorless, crystal
size 0.34!0.04!0.03 mm3, monoclinic, P21 [No. 4], aZ
˚
7.8949(3), bZ15.8034(6), cZ12.3838(5) A, bZ92.396(2)
8, VZ1543.73(10) A , ZZ2, DcalcZ1.210 Mg mK3, mZ
3
˚
0.234 mmK1, TZ100 K, 18212 reflections collected, 4405
independent reflections, 3171 reflections with IO2s(I),
qmaxZ23.258, 189 refined parameters, RZ0.0598, RwZ
0.1175, SZ1.032, largest diff. peak and holeZ
11. (a) Fu¨rstner, A.; Ruiz-Caro, J.; Prinz, H.; Waldmann, H.
J. Org. Chem. 2004, 69, 459–467. (b) Fu¨rstner, A. Eur. J. Org.
Chem. 2004, 943–958.
K3
˚
12. Brohm, D.; Philippe, N.; Metzger, S.; Bhargava, A.; Mu¨ller,
O.; Lieb, F.; Waldmann, H. J. Am. Chem. Soc. 2002, 124,
13171–13178.
0.479/K0.431 e A
.
13. (a) Bialy, L.; Waldmann, H. Angew. Chem. 2002, 114,
1819–1822. (b) Bialy, L.; Waldmann, H. Angew. Chem. Int.
Ed. 2002, 41, 1748–1751. (c) Bialy, L.; Waldmann, H. Chem.
Commun. 2003, 1872–1873.
Acknowledgements
We thank Dr. J. Kohno, Tanabe Seiyaku Co. Ltd, Japan, for
providing an authentic sample of TMC-69-6H for compari-
son and Dr. C. W. Lehmann for solving the X-ray structures.
Financial support by the DFG (Leibniz award to A. F.), the
Fonds der Chemischen Industrie and the Merck Research
Council is acknowledged with gratitude.
14. Preliminary communication: Fu¨rstner, A.; Feyen, F.; Prinz, H.;
Waldmann, H. Angew. Chem. 2003, 115, 5519–5522. Fu¨rstner,
A.; Feyen, F.; Prinz, H.; Waldmann, H. Angew. Chem. Int. Ed.
2003, 42, 5361–5364.
15. (a) Grynkiewicz, G.; Zamojski, A. Z. Naturforsch. B: Anorg.
Chem. Org. Chem. 1980, 35, 1024–1027. (b) Reetz,
M. T.; Mu¨ller-Starke, H. Liebigs Ann. Chem. 1983,
1726–1738.
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alkaloids that are not oxidized on nitrogen. For characteristic
members of these series see: (a) Militarinone A: Schmidt, K.;
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optical rotation of the authentic sample of 2 made available
to us, however, was lower than this reported value. When
corrected for the residual water in the sample, we record an
[a]20zC99 (cZ0.18, MeOH). The data of our analytically
pure synthetic compounds are as follows: (17R)-2. [a]20ZC
102.3 (cZ0.6, MeOH); (17S)-2. [a]20ZC88.6 (cZ0.95,
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