P2′-truncated BACE-1 inhibitors with a novel hydroxethylene-like core

Article abstract of DOI:10.1016/j.ejmech.2009.10.041

Highly potent BACE-1 protease inhibitors derived from a novel hydroxyethylene-like core structure were recently developed by our group using X-ray crystal structure data and molecular modelling. In a continuation of this work guided by molecular modelling we have explored a truncated core motif where the P2′ amide group is replaced by an ether linkage resulting in a set of alkoxy, aryloxy and alkylaryl groups, with the overall aim to reduce molecular weight and the number of amide bonds to increase permeability and bestow the inhibitors with drug-like features. The most potent of these inhibitors displayed a BACE-1 IC₅₀ value of 140 nM. The synthesis of these BACE-1 inhibitors utilizes readily available starting materials, furnishing the target compounds in good overall yields.

Full text of DOI:10.1016/j.ejmech.2009.10.041

European Journal of Medicinal Chemistry 45 (2010) 542–554
Contents lists available at ScienceDirect
European Journal of Medicinal Chemistry
journal homepage: http://www.elsevier.com/locate/ejmech
Original article
P20-truncated BACE-1 inhibitors with a novel hydroxethylene-like core
Jenny Adrian Meredith a, Catarina Bjo¨rklund a, Hans Adolfsson a, Katarina Jansson b, Anders Hallberg c,
,b,
Åsa Rosenquist b, Bertil Samuelsson a
*
a Department of Organic Chemistry, Arrhenius Laboratory, Stockholm University, SE-106 91 Stockholm, Sweden
b Medivir AB, PO Box 1086, SE-141 22 Huddinge, Sweden
c Department of Medicinal Chemistry, BMC, Uppsala University, Box 574, SE-751 23 Uppsala, Sweden
a r t i c l e i n f o
a b s t r a c t
Article history:
Received 2 July 2009
Received in revised form
27 September 2009
Accepted 22 October 2009
Available online 3 November 2009
Highly potent BACE-1 protease inhibitors derived from a novel hydroxyethylene-like core structure were
recently developed by our group using X-ray crystal structure data and molecular modelling. In
a continuation of this work guided by molecular modelling we have explored a truncated core motif
where the P20 amide group is replaced by an ether linkage resulting in a set of alkoxy, aryloxy and
alkylaryl groups, with the overall aim to reduce molecular weight and the number of amide bonds to
increase permeability and bestow the inhibitors with drug-like features. The most potent of these
inhibitors displayed a BACE-1 IC50 value of 140 nM. The synthesis of these BACE-1 inhibitors utilizes
readily available starting materials, furnishing the target compounds in good overall yields.
Ó 2009 Elsevier Masson SAS. All rights reserved.
Keywords:
BACE-1
BACE
b-Secretase
Asp2
Memapsin
PI
1. Introduction
Mice genetically deficient in BACE-1 show no Ab build-up in the
brain and are healthy and fertile [11–13]. These data together
suggest that inhibition of BACE-1 would be a suitable therapeutic
approach for AD. Intense research efforts are under way to identify
safe and efficacious drugs based on selective BACE-1 inhibition in
both the pharmaceutical industry and in academia [14–19].
As BACE-1 is a member of the aspartic protease family, most
inhibitors in progress incorporate a non-cleavable transition-state
isostere motif such as hydroxyethylamine (HEA), hydroxyethylene
(HE), statine, or norstatine pseudopeptide backbone frameworks,
where a hydroxy group is the key element [20]. Several research
groups have reported on promising BACE-1 inhibitors containing
substituted isophthalamides in the P2-P3 position and Vacca and
co-workers have described 5-substituted isophthalamides coupled
to hydroxyethylamine (HEA) cores that furnish potent and selective
BACE-1 inhibitors [21].
Previously we have demonstrated that a statin motif incorpo-
rating a methyl-aryloxy or methyl-benzyloxy P1 substituent with
a 5-substituted isophthalamide (A) [21] in the P2-P3 position
furnishes highly potent BACE-1 inhibitors [22]. We have also
recently disclosed a novel HE central core [23] having a methoxy
P10 substituent which renders BACE-1 inhibitors with higher
potency and improved BACE-1/cathepsin D selectivity, i.e. inhibitor
1 (Fig. 1) displaying an IC50 value of 0.32 nM. In the same work we
Alzheimer’s disease (AD) is a neurodegenerative, progressive
and ultimately fatal disorder afflicting approximately 40 percent of
the population over 80 years, with over 30 million people suffering
from AD worldwide [1,2]. In addition to the disease burden of the
individuals and social impact on families and society, the financial
cost to society is staggering, estimated at over 100 billion USD per
year in the US alone [3,4].
AD is characterized by the build-up of insoluble amyliod plaques
and neurofibrillary tangles in the brain tissue [5]. Amyloid plaque is
mainly constituted of aggregated A
with A (1-42) being the major component [6,7]. These peptides are
the cleavage products of amyloid precursor protein (APP), a trans-
membrane protein, by the action of BACE-1 ( -site APP-cleaving
enzyme) and -secretase. Invitro, the aggregation of A peptides has
shown to influence neurotoxicity [8,9]. In neuritic plaques found in
the brain of AD patients amyloid- 42 (A 1-42) dominates [10].
b(1-40) and Ab(1-42) peptides
b
b
g
b
b
b
* Corresponding author. Department of Organic Chemistry, Arrhenius Laboratory,
Stockholm University, SE-106 91 Stockholm, Sweden. Tel.: þ46 8 54683104; fax:
þ46 8 54683199.
E-mail address: bertil.samuelsson@medivir.com (B. Samuelsson).
0223-5234/$ – see front matter Ó 2009 Elsevier Masson SAS. All rights reserved.
doi:10.1016/j.ejmech.2009.10.041

J. Adrian Meredith et al. / European Journal of Medicinal Chemistry 45 (2010) 542–554
543
Scheme 2. Reagents and conditions: (i) NaH, RX, QI, DMF.
2. Chemistry
The lactol 4 (Scheme 1), synthesized from
D
-glucose in 8-steps
according to the literature procedure [24–30], was reduced with
LiBH4, rendering diol 3 in 95% yield. Compound 3 was activated
with dibutyl tinoxide, and regioselectively benzylated on the
primary alcohol position using QI and benzyl bromide to generate
5a (Scheme 1) in 38% yield. As selective methylation or allylation
employing this tin activation procedure only provided low yields
(10% or less) of the desired primary substituted hydroxyl group,
a different strategy was employed.
Treating diol 3 with sodium hydride and the appropriate organo
halides yielded the ethers 5b–h (Scheme 2), preferentially
substituted at the primary hydroxyl group, in 14–51% yields. The
undesired products, di-substituted and mono-substituted at the
secondary hydroxyl group, were formed in low yields, 2–14%, and
were readily separated by column chromatography.
Fig. 1. Lead compound 1 and target compound 2l.
have also shown that it is possible to truncate between the P20 and
P30 position and still maintain activity, although these compounds
are substantially less potent than lead compound 1.
For the conversion to the phenoxy ether, phenyl amines and
benzyl amine a different synthesis strategy was employed. The
secondary hydroxyl group was first protected as a p-methoxy
benzyl (PMB) ether using a selective reductive opening of the
p-methoxy-benzylidene acetal protected diol 6 [31]. The intro-
duction of p-methoxy-benzylidene acetal (Scheme 3) was per-
formed using anisaldehyde dimehtylacetal and p-TsOH in DMF
under reduced pressure to furnish compound 6 in 98% yield. Using
the described conditions for selective opening rendering the
unsubstituted primary hydroxyl (i.e. NaCNBH3, TMSCl, MeCN),
products 7 and 8 were delivered in 43% and 42% yield, respectively
[31]. The yield of compound 8 was not as high as expected, possibly
due to differences between a seven membered ring acetal and a six
membered ring acetal.[31] When using the alternative method
described to generate the corresponding PMB-protected primary
We now describe an approach to improve on the drug-like
properties of this new template by both reducing molecular weight
and decreasing the number of amide bonds (Fig. 1) rendering
a template that could easily could be varied in the P20 position,
allowing for the introduction of different ethers as well as
secondary amines.
The most potent of these inhibitors, the p-methoxy benzyl
(PMB) ether 2l, (Fig. 1) displayed a BACE-1 IC50 value of 140 nM.
In this study we report on the synthesis of BACE-1 inhibitors,
which are developments of the previously described P10-methoxy
hydroxethylene-like scaffold, having more drug-like features
although demonstrating somewhat lower potency. Further
modelling and preparative work to explore this truncated template
in BACE-1 inhibitor design is currently under way and will be
reported elsewhere.
Scheme 1. Reagents and conditions: (i): LiBH4, THF, 0 ꢀC; (ii) dibutyl tinoxide, toluene, reflux; (iii) QI, BnBr, 90 ꢀC.

544
J. Adrian Meredith et al. / European Journal of Medicinal Chemistry 45 (2010) 542–554
Scheme 5. Reagents and conditions: (i): 2-Nitro-N-phenyl-benzenesulfonamide or N-
(4-fluoro-phenyl)-2-nitro-benzenesulfonamide, DIAD, Ph3P, DCM; (ii): PhSH, K2CO3,
MeCN, 50 ꢀC; (iii) TFA 10% in DCM, 30 min.
with carboxylic acid A, the nosyl protecting group was not removed
until after coupling of the P2–P3 fragment. In order to confirm the
stereochemical preference of the secondary hydroxyl group, which
interacts with the catalytic Asp228 and Asp32 of BACE-1, the
hydroxyl epimer was also prepared. The secondary hydroxyl group
of 14 was thus inverted using Mitsunobu conditions, DIAD, tri-
phenylphosphine and p-nitro benzoic acid, followed by benzoate
saponification in 0.1 M NaOMe to deliver the hydroxyl epimer 15 in
51% yield over the two steps.
Reduction of the azide group in compounds 3, 5b–e, 8 and 12a
(Scheme 7) was achieved by hydrogenolysis providing the corre-
sponding amines which were coupled directly without prior puri-
fication with carboxylic acid A using PyBOP [34] and DIPEA in DCM
furnishing 2a–g in yields ranging from 39 to 85% over the two steps.
The azide group in 5a, 5f–h, 10 and 12b was reduced into the cor-
responding amines employing Staudinger conditions [35] i.e. Ph3P
in methanol containing a few drops of water. The amines were used
in the subsequent step without further purification and coupled to
carboxylic acid A, using the same procedure vide supra, to give
products 2h–m in 39–92% yield.
Azides 14 and 15 (Scheme 8) were also reduced into its corre-
sponding amines employing Staudinger conditions, coupled to
carboxylic acid A, to obtain 16 and 17 in 90% and 92% yield,
respectively. Compounds 16 and 17 were treated with thiophenol
and potassium carbonate to deliver 2n and 2o in 50% and 52% yield,
respectively.
Scheme 3. Reagents and conditions: (i): Anisaldehyde dimehtylacetal, p-TsOH, DMF,
reduced pressure, 50 ꢀC; (ii) NaCNBH3, TMSCl, MeCN, 0 ꢀC.
hydroxyl group (i.e. NaCNBH3, TFA, DMF) our results were in
accordance with those reported and generated almost exclusively
the product 7 [31].
Compound 7 could readily be separated from the desired
compound 8 and subsequently converted back to diol 3 in 90% yield
by treatment with 10% TFA in DCM. Compound 8 was converted
into its corresponding phenoxy ether 9 (Scheme 4) in 63% yield
employing Mitsunobu conditions after which the PMB group was
cleaved using DDQ to generate 10 in 89% yield.
Using Mitsunobu conditions (i.e. Ph3P, DIAD and DCM) the
nosylates [32] 2-nitro-N-phenyl-benzenesulfonamide and N-(4-
fluoro-phenyl)-2-nitro-benzenesulfonamide, were reacted with
compound 8 to deliver its corresponding nosylated phenyl amine
and 4-fluoro phenyl amine, which were subjected to thiophenol
and potassium carbonate cleavage of the nosyl activating group to
obtain 11a and 11b (Scheme 5) in 74% and 50% yield (over two
steps), respectively. The PMB-group was subsequently removed
using 10% TFA in DCM to obtain 12a and 12b in 84% and 99% yield,
respectively.
Compound 8 was converted into the nosylated benzyl amine 13
in 61% yield (Scheme 6) by Mitsunobu coupling with the nosylated
benzyl amine (N-(benzyl)-2-nitro-benzenesulfonamide) [33]. This
was followed by cleavage of the PMB-group using DDQ to generate
14 in 72% yield. To avoid undesired coupling of the benzyl amine
To evaluate two alternative P2-P3 groups, the azide moiety in
compound 7 was selectively reduced by hydrogenolysis (Scheme 9)
and coupled with carboxylic acids B [36] and C [37] to furnish the
final products 18 and 19 in 66 and 76% yield, respectively.
3. Structure activity relationships
In a previous study we have demonstrated that an HE motif
incorporating a methyl-aryloxy P1 substituent and a methoxy P10
substituent coupled to isophathalamide A in the P2-P3 position
furnish highly potent and cathepsin D selective BACE-1 inhibitors,
i.e. inhibitor 1 [23] displaying an IC50 value of 0.32 nM. With the
Scheme 6. Reagents and conditions: (i): N-(benzyl)-2-nitro-benzenesulfonamide,
DIAD, Ph3P, DCM; (ii): DDQ, DCM/H2O 19:1; (iii): p-nitro benzoic acid, DIAD, Ph3P, THF;
Scheme 4. Reagents and conditions: (i): Phenol, Ph3P, DIAD, DCM; (ii) DDQ, DCM/H2O
19:1.
(iv): NaOMe 0.1 M, DOWEX Hþ
.

J. Adrian Meredith et al. / European Journal of Medicinal Chemistry 45 (2010) 542–554
545
Scheme 9. Reagents and conditions: (i): H2, Pd/C, MeOH; (ii): PyBOP, DIPEA, B or C,
DCM.
Scheme 7. Reagents and conditions: (i): H2, Pd/C, MeOH; (ii): Ph3P, H2O, MeOH; (iii):
PyBOP, DIPEA, A, DCM.
In order to explore the impact on varying the P2-P3 substitu-
ents, the azide in compound 7 was reduced and coupled with acids
B and C delivering compounds, 18 and 19 (entries 16 and 17, Table
1). Both showed an approximately tenfold decrease in potency
compared to 2l (entry 12, Table 1). Although all three carboxylic
acids employed has shown similar impact on potency in previous
studies, it indicates that the S2-S3-pockets are sensitive for the
conformal changes that appear when the amide on the prime side
is lost and the interactions with the S20 pocket are changed. The
phenyl amine 2g is four times as potent as phenyl ether 2f, this may
be due to positive interaction between the nitrogen and the
carbonyl of Gly34. Our results indicate that the interactions to the
amides are of great importance, which suggests that, a major
change in the P20 position is required to regain activity and that
likely new interactions are also required to make up for this loss.
Work towards this end is currently under way.
aim to bestow this inhibitor class with more advantageous drug-
like properties the two prime side amides having one side-chain
and three backbone hydrogen bonds to the active site of BACE-1
were removed. All targets were screened against BACE-1 and the
IC50 values are shown in Table 1. Modelling (Fig. 2) suggests that
this truncation is likely to result in less active inhibitors unless an
optimal P20 group having efficient interactions in the S20 pocket of
BACE-1 can be introduced.
The shortening of the compounds will also position the capping
P20 group in the buried S20 pocket rather than in the shallow S30
pocket. Inhibitor 2c with an IC50 of 484 nM (entry 3, Table 1) is
exhibiting similar interactions as inhibitor 1 in the S20 pocket, and
with the loss of the amides and the P30 interactions this compound is
losing activity. Inhibitors 2k and 2l with IC50 values of 243 nM and
140 nM, respectively, (entry 11 and 12, Table 1) are having more
extensive interactions in S20, including aromatic stacking with Tyr71
and Arg128, increasing the activity compared to 2c. According to the
3D structure the p-methoxy substituent in 2l makes additional close
contact interactions with Ser36, Asn37 and Ile126 in the bottom of
the S20 pocket, making it twice as active as 2k. The meta-methoxy-
benzyl compound 2j is loosing activity due to the restrictions in
available space for a m-methoxy group in the S20 pocket. Overall
these inhibitors demonstrate a narrow SAR and relative modest
potency indicating that additional interactions, i.e. a larger S10 group
or interacting with Tyr198 in proximity to the S20 pocket, might be
advantageous for delivering high potency inhibitors.
4. Conclusions
Based on a novel hydroxyethylene-like core structure recently
developed by our group, using X-ray crystal structure data and
molecular modelling, a new series of drug-like truncated BACE-1
inhibitors were designed and synthesized where 2c, 2k and 2l
showed potency in the nanomolar range. These inhibitors are
considerable less peptidic than those previously reported by our
group but they also display less potency compared with the potent
BACE-1 inhibitor 1. We have showed that the interactions in the S20
and S30-pocket as well as the amide linkage is of key importance for
potency and that loss or great change of these interactions must be
compensated in other ways. We believe that truncated, less peptide
like inhibitors is an area that need to be investigated further.
5. Experimental section
5.1. Protease enzyme assays
The BACE-1 assay was performed as previously described [22].
5.1.1. Modelling
Binding modes and enzyme interactions of inhibitors 2c, 2k and
2l were investigated using GOLD docking [39] with visualisation
using Sybyl [40]. The inhibitors were docked in Gold 200 times each
using constraints for hydrogen bonding to the catalytic aspartic
acids (Asp32 and Asp228) and to the backbone carbonyls of Gly34
Scheme 8. Reagents and conditions: (i): Ph3P, H2O, MeOH; (ii): PyBOP, DIPEA, A, DCM;
(iii): PhSH, K2CO3, MeCN, 50 ꢀC.

546
J. Adrian Meredith et al. / European Journal of Medicinal Chemistry 45 (2010) 542–554
Table 1
BACE-1 inhibitory activities.
OH
O
O
F
H
N
R2
R1
F
Entry
Cpd.
R1
R2
OH
IC50 (mM)
O
S
O
O
O
N
OH
H
N
1
2a
2b
(S)-OH
>10
O
O
S
N
N
OMe
2
3
H
N
(S)-OH
(S)-OH
>10
O
O
O
O
O
S
O
O
H
N
2c
2d
2e
0.48
O
O
O
O
O
O
S
N
N
H
N
4
5
(S)-OH
(S)-OH
1.86
O
O
O
S
O
CF3
H
N
n.d.
O
S
O
O
N
N
O
H
N
6
2f
(S)-OH
6.50
O
O
O
S
O
O
H
N
H
N
7
8
2g
2h
(S)-OH
(S)-OH
1.50
2.03
O
S
O
O
O
N
H
N
O

J. Adrian Meredith et al. / European Journal of Medicinal Chemistry 45 (2010) 542–554
547
Table 1 (continued)
Entry
Cpd.
R1
R2
OH
IC50 (mM)
O
S
O
O
N
N
N
N
N
N
N
F
H
N
O
9
2i
(S)-OH
>10
O
O
O
O
O
O
O
O
S
O
O
O
H
N
10
11
12
13
14
15
2j
O
(S)-OH
(S)-OH
(S)-OH
(S)-OH
(S)-OH
(R)-OH
>10
0.24
0.14
1.50
>10
>10
O
S
O
O
O
H
N
2k
O
S
O
O
O
H
N
O
2l
O
S
H
N
O
O
H
N
2m
2n
2o
F
O
S
O
O
H
N
H
N
O
S
O
O
H
N
H
N
O
S
O
N
O
O
N
O
O
16
17
18
19
(S)-OH
(S)-OH
1.80
1.50
N
O
O
S
N
O
N
O

548
J. Adrian Meredith et al. / European Journal of Medicinal Chemistry 45 (2010) 542–554
(d, 1JCF ¼ 246.6 Hz), 160.2, (t, 3JCF ¼ 13.9 Hz), 98.6, (d, 2JCF ¼ 29.1 Hz,
2C), 97.1, (t, 2JCF ¼ 25.7 Hz), 80.4, 69.4, 64.6, 62.8, 57.1, 34.8; MS (ESI)
m/z 340.1 ([M þ Na]þ calcd for C13H17F2N3NaOþ4 340.1)
5.1.3.2. (2S,3S,5R)-2-azido-6-benzyloxy-1-(3,5-difluoro-phenoxy)-5-
methoxy-hexan-3-ol (5a). Under argon atmosphere dibutyl tin-
oxide (85 mg, 0.34 mmol) was added to a stirred solution of 3
(82 mg, 0.26 mmol) in dry toluene (3 mL). The reaction was
refluxed for 2 h with a Dean–Stark trap. After 2 h benzyl bromide
(61 mL, 0.52 mmol) and tert-butyl ammonium iodide (126 mg,
0.34 mmol) were added. The reaction mixture was refluxed 1 h
before it was concentrated and purified by silica column chro-
matography (toluene–ethyl acetate, gradient 30:1–5:1) to give 5a
20
in 38% yield (40 mg, 98
(400 MHz, CDCl3)
m
mol). [
a
]
D
þ1.8 (c 1.0, MeOH); 1H NMR
d
7.39–7.28 (m, 5H), 6.50–6.41 (m, 3H), 4.56 (d,
2H, J ¼ 2.8 Hz), 4.24 (dd, 1H, J ¼ 4.1, 9.8 Hz), 4.13 (dd, 1H, J ¼ 8.0,
9.7 Hz), 4.04 (m, 1H), 3.72 (d, 1H, J ¼ 2.4 Hz), 3.66 (m, 2H), 3.55 (d,
2H, J ¼ 4.5 Hz), 3.45 (s, 3H), 1.94–1.77 (m, 2H); 13C NMR (100 MHz,
Fig. 2. Models of compounds 2c (green), 2k (yellow), and 2l (orange) super-
impositioned with an X-ray crystal structure of inhibitor 1 (white) co-crystallized with
BACE-1 [38]. Tyr71 and Arg128 of the enzyme are included for clarity.
CDCl3)
d
163.9 (d, 1JCF ¼ 246.5 Hz), 163.7 (d, 1JCF ¼ 246.5 Hz), 160.3
3
2
(t, JCF ¼ 13.6 Hz), 137.8, 128.6, 128.0, 127.9, 98.6 (d, JCF ¼ 28.2 Hz,
2
2C), 97.1 (t, JCF ¼ 25.7 Hz), 80.1, 73.7, 71.1, 70.5, 68.6, 64.3, 57.6,
and Gly230. The binding conformations were evaluated using
GoldScore. The X-ray crystal structure of inhibitor 1 [38] co-crys-
tallized with BACE-1 was used as the starting point for the
modelling and docking studies.
35.5; MS (ESI) m/z 430.2 ([M þ Na]þ calcd for C20H23F2N3NaOþ4
430.2)
5.1.3.3. 2(S,3S,5R)-2-azido-1-(3,5-difluoro-phenoxy)-5,6-dimethoxy-
hexan-3-ol (5b). NaH (60% in mineral oil) (15 mg, 0.37 mmol,
1.2 equiv) was added to a stirred solution of diol 3 (99 mg,
0.31 mmol, 1 equiv) in dry DMF (2 mL) at 0 ꢀC. After 30 min methyl
5.1.2. General methods
All glassware were dried over an open flame before use in
connection with an inert atmosphere. Concentrations were per-
formed under reduced pressure at <40 ꢀC (bath temperature). Thin
layer chromatography was performed using Merck silica gel 60 F-
254 plates with detection by UV, charring with 8% sulphuric acid or
ammonium molybdate (100 g): Ce(IV)sulphate (2 g): sulphuric acid
(10%, 2L). Column chromatography was performed on silica (0.035–
0.070 mm). NMR spectra were recorded at 25 ꢀC on a Varian
(400 MHz) or on a Bruker (400 MHz or 500 MHz) instrument using
iodide (29
mL, 0.47 mmol, 1.5 equiv) and tert-butyl ammonium
iodide (11 mg, 31
mmol, 0.1 equiv) were added. The reaction was
allowed to reach r t and then left stirring for an additional 16 h. The
reaction was quenched with MeOH, concentrated and purified by
silica column chromatography (toluene–ethyl acetate, gradient
30:1–5:1) to give title compound 5b in 23% yield (24 mg, 72
m
mol).
20
[a
]
ꢂ3.4 (c 0.5, MeOH); 1H NMR (400 MHz, CDCl3)
d 6.50–6.41 (m,
D
3H), 4.26 (dd, 1H, J ¼ 4.1, 9.8 Hz), 4.15 (dd, 1H, J ¼ 8.0, 9.8 Hz), 4.05
(m, 1H), 3.76 (d, 1H, J ¼ 2.4 Hz), 3.69 (m, 1H), 3.60 (m, 1H), 3.49 (m,
2H). 3.46 (s, 3H), 3.40 (s, 3 H), 1.86 (m, 2H); 13C NMR (100 MHz,
the solvent residual peak (CDCl3 1H
d
¼ 7.26 and 13C
d
¼ 77.16 or
CD3OD-d4 1H
d
¼ 3.31 and 13C ¼ 49.0) as standard. Unless stated
otherwise, all materials were obtained from commercial suppliers
and used without further purification. DCM was refluxed over CaH2
and distilled onto 4 Å MS before use. Compound 2a–o, 18 and 19
were further purified before the biological testing using preparative
RP-HPLC, consisted of Waters 2767 auto-injector and fraction
collector, Waters 996 photodiode array detector, and Micromass
ZQ2000 mass detector (operated in þESI). The preparative reversed
CDCl3,)
d
163.9 (d, 1JCF ¼ 246.5 Hz), 163.7 (d, 1JCF ¼ 246.5 Hz), 160.3
(t, 3JCF ¼ 13.9 Hz), 98.6 (d, 2JCF ¼ 28.4 Hz, 2C), 97.1 (t, 2JCF ¼ 25.9 Hz),
79.8, 73.8, 70.2, 68.7, 64.5, 59.5, 57.5, 35.5; MS (ESI) m/z 354.1
([M þ Na]þ calcd for C14H19F2N3NaOþ4 354.1)
5.1.3.4. (2S,3S,5R)-2-azido-1-(3,5-difluoro-phenoxy)-6-isobutoxy-5-
methoxy-hexan-3-ol (5c). Title compound 5c was obtained from 3
20
phase column was an ACE C8, 21 ꢁ100 mm, 5
m
m, 100 A from ACE
as described for 5b, in 14% yield (18 mg, 48
m
mol). [
a
]
ꢂ3.7 (c 1.00,
D
(UK) and the mobile phases were based on water/acetonitrile
containing 0.1%ammonium acetate. Optical rotations were
measured at r t on a Perkin Elmer 341 polarimeter using a 10 cm,
1 mL cell. Mass spectra were recorded with a Bruker micrOTOF 125
spectrometer.
MeOH); 1H NMR (400 MHz, CDCl3)
d 6.50–6.41 (m, 3H), 4.26 (dd,
1H, J ¼ 4.2, 9.8 Hz), 4.15 (dd, 1H, J ¼ 8.0, 9.8 Hz), 4.06 (m, 1H), 3.85
(d, 1H, J ¼ 2.2 Hz), 3.68 (m, 1H), 3.32 (m, 1H), 3.50 (m, 2H). 3.46 (s,
3H), 3.22 (m, 2H), 1.92–2.78 (m, 3H), 0.91 (d, 3H, J ¼ 1.3 Hz), 0.89 (d,
3H, J ¼ 1.3 Hz); 13C NMR (100 MHz, CDCl3,)
d
163.9 (d,
1
3
1JCF ¼ 246.3 Hz), 163.7 (d, JCF ¼ 246.3 Hz), 160.3 (t, JCF ¼ 13.7 Hz),
2
2
5.1.3. Synthetic experiments
98.6 (d, JCF ¼ 28.5 Hz, 2C), 97.1 (t, JCF ¼ 25.9 Hz), 80.0, 78.8, 72.4,
70.3, 68.7, 64.4, 57.7, 35.6, 28.5, 19.5, 19.4; MS (ESI) m/z 374.2
([M þ H]þ calcd for C17H26F2N3Oþ4 374.2)
5.1.3.1. (2R,4S,5S)-5-azido-6-(3,5-difluoro-phenoxy)-2-methoxy-
hexane-1,4-diol (3). Under argon atmosphere LiBH4 (200 mg,
9.22 mmol) was added to a stirred solution of 4 (1.45 g, 4.61 mmol)
in dry THF (20 mL) at 0 ꢀC. After 1 h the reaction mixture was
quenched with water, extracted with ethyl acetate, dried over
Na2SO4 and concentrated. After purification by silica gel chroma-
5.1.3.5. (2S,3S,5R)-2-azido-6-cyclopropylmethoxy-1-(3,5-difluoro-phe-
noxy)-5-methoxy-hexan-3-ol (5d). Title compound 5d was
obtained from 3 as described for 5b, in 43% yield (47 mg,
20
tography (toluene–ethyl acetate, gradient 20:1–5:1) compound 3 in
0.13 mmol). [
d
a]
ꢂ5.1 (c 1.0, MeOH); 1H NMR (400 MHz, CDCl3)
D
20
95% yield (1.39 g, 4.38 mmol). [
(400 MHz, CDCl3)
a
]
D
ꢂ5.8 (c 1.00, MeOH); 1H NMR
6.50–6.39 (m, 3H), 4.26 (dd, 1H, J ¼ 4.1, 9.7 Hz), 4.16 (dd, 1H,
d
6.50–6.41 (m, 3H), 4.26 (dd, 1H, J ¼ 4.1, 9.7 Hz),
J ¼ 8.0, 9.8 Hz), 4.07 (m, 1H), 3.92 (d, 1H, J ¼ 2.3 Hz), 3.68 (m, 1H),
3.62 (m, 1H), 3.55 (m, 2H). 3.45 (s, 3H), 3.38–3.26 (m, 2H), 1.86 (m,
2H), 1.05 (m, 1H), 0.55(m, 2H), 0.20 (m, 2H); 13C NMR (100 MHz,
4.15 (dd, 1H, J ¼ 8.0, 9.8 Hz), 4.05 (m, 1H), 3.88 (m, 1H), 3.71 (m, 1H),
3.36–3.23 (m, 2H). 3.45 (s, 3H), 2.01–1.90 (m, 1H), 1.80 (m, 1H), 1.64
(bs,1H); 13C NMR (100 MHz, CDCl3)
d
163.9 (d,1JCF ¼ 246.6 Hz),163.7
CDCl3,)
d
163.9 (d, 1JCF ¼ 246.8 Hz), 163.7 (d, 1JCF ¼ 246.8 Hz), 160.3

J. Adrian Meredith et al. / European Journal of Medicinal Chemistry 45 (2010) 542–554
549
(t, 3JCF ¼ 13.6 Hz), 98.6 (d, 2JCF ¼ 28.4 Hz, 2C), 97.1 (t, 2JCF ¼ 25.9 Hz),
79.9, 76.5, 71.6, 70.0, 68.7, 64.5, 57.5, 35.8, 10.5, 31.2, 31.1; MS (ESI)
m/z 394.2 ([M þ Na]þ calcd for C17H23F2N3NaOþ4 394.2)
drops of TEA were added and the reaction mixture was concen-
trated and purified by silica column chromatography (toluene–
ethyl acetate, gradient 100:1–10:1) to give the product 6 in 98%
yield (382 mg, 0.88 mmol). 1H NMR (400 MHz, CDCl3)
d 7.40 (m,
5.1.3.6. (2S,3S,5R)-2-azido-1-(3,5-difluoro-phenoxy)-5-methoxy-6-
(4,4,4-trifluoro-butoxy)-hexan-3-ol (5e). Title compound 5e was
obtained from 3 as described for 5b, in 36% yield (48 mg,
2H), 6.89 (m, 2H), 6.38 (m, 1H), 6.07 (m, 2H), 5.77 (s, 1H), 4.33 (m,
1H), 4.23 (m, 1H), 3.86 (m, 1H), 3.78 (s, 3H) 3.55 (m, 3H), 3.48 (s,
3H), 3.06 (dd, 1H, J ¼ 3.6, 9.2 Hz), 2.06 (m, 1H), 1.90 (m, 1H); 13C
20
1
011 mmol). [
d
a
]
ꢂ4.1 (c 1.0, MeOH); 1H NMR (400 MHz, CD3OD)
NMR (100 MHz, CDCl3)
d
3
163.9 (d, JCF ¼ 246.6 Hz), 163.7 (d,
D
6.63–6.51 (m, 3H), 4.27 (dd, 1H, J ¼ 4.0, 10.0 Hz), 4.17 (dd, 1H,
1JCF ¼ 246.6 Hz), 160.1 (t, JCF ¼ 13.8 Hz), 159.9, 131.1, 128.0, 127.7,
2
2
J ¼ 7.8, 10.0 Hz), 3.93 (m, 1H), 3.76 (m, 1H), 3.61–3.49 (m, 5H). 3.41
113.7, 98.6 (d, JCF ¼ 28.8 Hz, 2C), 97.3 (t, JCF ¼ 25.7 Hz), 77.4, 76.5,
72.0, 67.9, 63.8, 62.8, 56.5, 55.5, 36.6; MS (ESI) m/z 458.2 ([M þ Na]þ
calcd for C21H23F2N3NaOþ5 458.2)
(s, 3H), 2.30–2.17 (m, 2H), 1.81 (m, 4H); 13C NMR (100 MHz, CD3OD)
1
1
d
165.3 (d, JCF ¼ 244.9 Hz), 165.1 (d, JCF ¼ 244.9 Hz), 162.1 (t,
3JCF ¼ 13.9 Hz), 129.0 (d, 1JCF ¼ 274.0 Hz), 99.5 (d, 2JCF ¼ 29.0 Hz, 2C),
97.4 (t, 2JCF ¼ 26.4 Hz), 79.0, 73.0, 70.6, 70.2, 69.2, 66.1, 57.5, 31.5 (dd,
2JCF ¼ 28.8, 86.7 Hz), 31.0, 23.5 (dd, 3JCF ¼ 2.9, 8.9 Hz); MS (ESI) m/z
450.1 ([M þ Na]þ calcd for C17H22F5N3NaOþ4 450.1)
5.1.3.11. (2S,3S,5R)-2-azido-1-(3,5-difluoro-phenoxy)-5-methoxy-6-
(4-methoxy-benzyloxy)-hexan-3-ol (7) and (2R,4S,5S)-5-azido-6-
(3,5-difluoro-phenoxy)-4-(4-methoxy-benzyloxy)-hexan-1-ol (8). A
solution, kept at 0 ꢀC, of Me3SiCl (0.48 mL, 3.85 mmol) in MeCN
(3 mL) was added drop wise to a stirred mixture of 6 (279 mg,
0.64 mmol), NaCNBH3 (242 mg, 3.85 mmol) and 3 Å molecular
sieves in MeCN (7.5 mL). The reaction mixture was stirred at 0 ꢀC for
30 min and then allowed to reach r t before it was filtrated through
Celite and evaporated. The residue was dissolved in DCM and
washed with NaHCO3(aq), the organic phase was dried over
Na2SO4, evaporated and purified by silica column chromatography
(toluene–ethyl acetate, gradient 60:1–3:1) to give the products 7 in
5.1.3.7. (2S,3S,5R)-6-allyloxy-2-azido-1-(3,5-difluoro-phenoxy)-5-
methoxy-hexan-3-ol (5f). Title compound 5f was obtained from 3
20
as described for 5b, in 51% yield (109 mg, 0.31 mmol). [
a]
ꢂ3.4 (c
D
1.0, MeOH); 1H NMR (400 MHz, CD3OD)
d
6.62–6.50 (m, 3H), 5.91
(m, 1H), 5.28 (m, 1H), 5.16 (m, 1H), 4.26 (dd, 1H, J ¼ 4.0, 10.0 Hz),
4.16 (dd, 1H, J ¼ 7.9, 10.0 Hz), 4.01 (m, 2H), 3.93 (m, 1H), 3.76 (m,
1H), 3.58 (m, 2H). 3.51 (m, 1H), 3.40 (s, 3H), 1.83 (m, 2H); 13C NMR
1
(100 MHz, CD3OD)
d
165.3 (d, JCF ¼ 244.9 Hz), 164.0 (d,
1JCF ¼ 244.9 Hz), 165.1 (t, JCF ¼ 14.0 Hz), 136.0, 117.3, 99.5 (d,
2JCF ¼ 29.2 Hz, 2C), 97.4 (t, 2JCF ¼ 26.4 Hz), 79.0, 73.3, 72.3, 70.2, 69.2,
66.1, 57.5, 36.3; MS (ESI) m/z 380.1 ([M þ Na]þ calcd for
C16H21F2N3NaOþ4 380.1)
43% yield (121 mg, 0.27 mmol) and 8 in 42% yield (118 mg,
1
20
0.26 mmol,). (7) [
a
]
ꢂ1.0 (c 1.00, MeOH); 1H NMR (400 MHz,
D
CDCl3)
d
7.25 (m, 2H), 6.89 (m, 2H), 6.50–6.41 (m, 3H), 4.49 (m, 2H),
4.23 (dd, 1H, J ¼ 4.1, 9.7 Hz), 4.13 (dd, 1H, J ¼ 8.0, 9.6 Hz), 4.02 (m,
1H), 3.80 (s, 3H), 3.74 (d, 1H, J ¼ 2.3 Hz), 3.67 (m, 1H), 3.62 (m, 1H),
3.52 (m, 1H), 3.43 (s, 3H), 1.85 (m, 2H); 13C NMR (100 MHz, CDCl3)
5.1.3.8. (2S,3S,5R)-2-azido-1-(3,5-difluoro-phenoxy)-6-(4-fluoro-ben-
zyloxy)-5-methoxy-hexan-3-ol (5g). Title compound 5g was
1
1
d
163.9 (d, JCF ¼ 246.7 Hz), 163.7 (d, JCF ¼ 246.7 Hz), 160.3 (t,
obtained from 3 as described for 5b, in 29% yield (48 mg,
3JCF ¼ 13.6 Hz), 159.5, 129.8, 129.6, 114.0, 98.6 (d, 2JCF ¼ 28.6 Hz, 2C),
20
2
0.11 mmol). [
d
a]
þ0.2 (c 1.0, MeOH); 1H NMR (400 MHz, CD3OD)
97.1 (t, JCF ¼ 25.8 Hz), 80.0, 73.4, 70.7, 70.4, 68.6, 64.4, 57.6, 55.4,
D
7.35 (m, 2 H), 7.04 (m, 2H), 6.60–6.50 (m, 3H), 4.51 (m, 2H), 4.23
35.6; MS (ESI) m/z 460.2 ([M þ Na]þ calcd for C21H25F2N3NaOþ5
20
(dd, 1H, J ¼ 4.0, 10.1 Hz), 4.15 (dd, 1H, J ¼ 7.8, 10.1 Hz), 3.90 (m, 1H),
460.2). (8) [
a
]
þ1.7 (c 1.00, MeOH); 1H NMR (400 MHz, CDCl3)
D
3.74 (m, 1H), 3.61 (m, 2H), 3.54 (m, 1H), 3.40 (s, 3H), 1.85 (m, 2H);
d 7.25 (m, 2H), 6.87 (m, 2H), 6.47–6.34 (m, 3H), 4.57 (d, 1H,
1
13C NMR (100 MHz, CD3OD,)
d
165.3 (d, JCF ¼ 246.9 Hz), 165.1 (d,
J ¼ 11.3 Hz), 4.46 (d, 1H, J ¼ 11.3 Hz), 4.03 (m, 2H), 3.87 (m, 1H), 3.80
1JCF ¼ 246.9 Hz), 163.8 (d, JCF ¼ 245.4 Hz), 162.0 (t, JCF ¼ 13.6 Hz),
(s, 3H), 3.75 (m, 2H), 3.51 (m, 1H), 3.42 (m, 1H), 3.39 (s, 3H), 1.90 (m,
1
3
4
3
135.7 (d, JCF ¼ 3.1 Hz), 130.8 (d, JCF ¼ 8.2 Hz 2C), 116.0 (d,
2JCF ¼ 21.8 Hz, 2C), 99.5 (d, 2JCF ¼ 28.7 Hz 2C), 97.4 (t, 3JCF ¼ 26.3 Hz),
79.0, 73.5, 72.2, 70.2, 69.2, 66.1, 57.5, 36.3; MS (ESI) m/z 448.1
([M þ Na]þ calcd for C20H22F3N3NaOþ4 448.2)
2H); 13C NMR (100 MHz, CDCl3)
d
163.8 (d, 1JCF ¼ 246.6 Hz),163.6 (d,
3
1JCF ¼ 246.6 Hz), 160.1 (t, JCF ¼ 13.6 Hz), 159.7, 130.0, 129.6, 114.1,
2
2
98.6 (d, JCF ¼ 28.5 Hz, 2C), 97.1 (t, JCF ¼ 25.8 Hz), 78.2, 74.6, 71.9,
68.5, 63.3, 62.3, 57.2, 55.4, 31.6; MS (ESI) m/z 460.2 ([M þ Na]þ calcd
for C21H25F2N3NaOþ5 460.2)
5.1.3.9. (2S,3S,5R)-2-azido-1-(3,5-difluoro-phenoxy)-5-methoxy-6-
(3-methoxy-benzyloxy)-hexan-3-ol (5h). Title compound 5h was
5.1.3.12. (2S,3S,5R)-2-azido-1-(3,5-difluoro-phenoxy)-3-(4-methoxy-
obtained from 3 as described for 5b, in 24% yield (55 mg,
benzyloxy)-5-methoxy-6-phenoxy-hexane
(9). DIAD
(28
mL,
20
0.13 mmol). [
a
]
ꢂ4.3 (c 1.0, MeOH); 1H NMR (400 MHz, CD3OD)
0.14 mmol) was added drop wise at 0 ꢀC to a stirred solution of 8
(44 mg, 0.10 mmol), phenol (14.3 mg, 0.15 mmol) and Ph3P (37 mg,
0.14 mmol) in freshly distilled DCM (3 mL). The reaction mixture
was allowed to reach r t. After 16 h the reaction mixture was
D
d
7.23 (t, 1H, J ¼ 7.8 Hz), 6.91 (m, 2H), 6.82 (m, 1H), 6.60–6.50 (m,
3H), 4.51 (dd, 2H, J ¼ 4.1, 20.4 Hz), 4.22 (dd, 1H, J ¼ 4.0, 10.1 Hz), 4.14
(dd, 1H, J ¼ 7.8, 10.1 Hz), 3.87 (m, 1H), 3.77 (s, 3H), 3.73 (m, 1H), 3.60
(m, 2H), 3.54 (m, 1H), 3.40 (s, 3H), 1.85 (t, 2H, J ¼ 3.6 Hz); 13C NMR
evaporated and purified by column chromatography (toluene) to
1
20
(100 MHz, CD3OD)
d
165.2 (d, JCF ¼ 245.4 Hz), 165.1 (d,
give the product 9 in 63% yield (33 mg, 63
mmol). [
a
]
ꢂ3.6 (c 0.5,
D
1JCF ¼ 245.4 Hz), 162.0, (t, JCF ¼ 13.7 Hz), 161.3, 141.2, 130.4, 121.0,
114.3, 114.2, 99.5 (d, 2JCF ¼ 28.9 Hz 2C), 97.4 (t, 2JCF ¼ 26.3 Hz), 79.0,
74.2, 72.0, 70.2, 69.2, 66.2, 57.4, 55.6, 36.3; MS (ESI) m/z 460.2
([M þ Na]þ calcd for C21H25F2N3NaOþ5 460.2)
MeOH); 1H NMR (500 MHz, CDCl3)
d 7.32 (m, 5H), 7.0–6.51 (m, 4H),
3
6.48–6.35 (m, 3H), 4.59 (d, 1H, J ¼ 11.4 Hz), 4.45 (d, 1H, J ¼ 11.4 Hz),
4.09–3.92 (m, 3H), 3.86 (m, 2H), 3.79 (s, 3H), 3.72 (m, 1H), 3.62 (m,
1H), 3.47 (s, 3H), 2.11 (m, 1H), 1.96 (m, 1H); 13C NMR (125 MHz,
CDCl3)
d
164.6, 163.8 (d, 1JCF ¼ 246.9 Hz), 163.7 (d, 1JCF ¼ 246.9 Hz),
5.1.3.10. (4S,6R)-4-[(1S)-1-azido-2-(3,5-difluoro-phenoxy)-ethyl]-6-
methoxy-2-(4-methoxy-phenyl)-[1,3]dioxepane (6). Anisaldehyde
160.2, (t, 3JCF ¼ 13.6 Hz), 159.6, 158.7, 130.8, 129.8, 129.7, 121.3, 114.7,
114.0, 98.6 (d, 2C, 2JCF ¼ 28.6 Hz,), 97.1 (d, 2JCF ¼ 25.8 Hz), 76.2, 74.4,
71.7, 69.2, 68.6, 62.5, 57.8, 55.4, 32.4; MS (ESI) m/z 536.2 ([M þ Na]þ
calcd for C27H29F2N3NaO5 536.2)
dimethylacetal (228
mL, 1.34 mmol) and p-TsOH (8 mg, 45 mmol)
were added to a stirred solution of 3 (283 mg, 0.89 mmol) in freshly
distilled DMF (10 mL). The reaction mixture was heated to 50 ꢀC
under reduced pressure. The reaction was followed by TLC, after
approximately 1.5 h all starting material was consumed. A couple of
5.1.3.13. (2S,3S,5R)-2-azido-1-(3,5-difluoro-phenoxy)-5-methoxy-6-
phenoxy-hexan-3-ol (10). DDQ (12 mg, 0.05 mmol) was added at

550
J. Adrian Meredith et al. / European Journal of Medicinal Chemistry 45 (2010) 542–554
20
20
0 ꢀC to a stirred solution of 9 (20 mg, 39
10 mL). The reaction was allowed to reach r t then stirred for an
additional 3 h. The reaction mixture was washed with saturated
NaHCO3 (aq) and water, dried over Na2SO4, evaporated and purified
m
mol) in DCM:H20 (19:1,
(103 mg, 0.19 mmol). [
a
]
ꢂ5.9 (c 1.00, CHCl3); [
a
]
ꢂ5.9 (c 1.00,
D
D
CHCl3); 1H NMR (400 MHz, CDCl3)
d 7.24 (m, 2H), 6.88 (m, 4H), 6.53
(m, 2H), 6.48–6.35 (m, 3H), 4.52 (m, 2H), 3.99 (m, 2H) 3.87 (m, 1H),
3.79 (s, 3H), 3.88 (m, 1H), 3.58 (m, 1H), 3.38 (s, 3H), 3.20 (d, 1H,
by column chromatography (toluene–ethyl acetate, 50:1) to give
J ¼ 4.5, 12.5 Hz), 3.05 (d, 1H, J ¼ 5.3, 12.5 Hz), 1.95 (m, 2H); 13C NMR
20
1
the product 10 in 89% yield (14 mg, 35
mmol). [
a]
ꢂ3.6 (c 0.5,
(100 MHz, CDCl3)
d
163.9 (d, JCF ¼ 245.6 Hz), 163.7 (d,
D
MeOH); 1H NMR (400 MHz, CD3OD)
d
7.26 (m, 2H), 6.92 (m, 2H),
1JCF ¼ 245.6 Hz), 160.1 (t, 3JCF ¼ 13.7 Hz), 159.7, 156.1 (d,
4
6.56 (m, 3H), 4.27 (dd,1H, J ¼ 4.0,10.1 Hz), 4.18 (m,1H), 4.11 (m,1H),
1JCF ¼ 234.8 Hz), 145.5 (d, JCF ¼ 2.0 Hz), 130.0, 129.6, 115.8 (d, 2C
4.04 (m, 1H), 3.98 (m, 1H), 3.80 (m, 2H), 3.47 (s, 3H), 1.96 (m, 2H);
2JCF ¼ 22.4 Hz), 114.0 (d, 2C 3JCF ¼ 7.6 Hz), 98.6 (d, 2C 2JCF ¼ 20.3 Hz),
13C NMR (100 MHz, CD3OD)
d
165.3 (d, JCF ¼ 245.4 Hz), 165.1 (d,
97.1 (t, JCF ¼ 26.0 Hz), 76.3, 75.6, 71.9, 68.5, 62.3, 56.9, 55.4, 46.7,
1
2
1JCF ¼ 245.4 Hz), 162.0 (t, JCF ¼ 13.9 Hz), 160.2, 130.5, 122.0, 115.6,
32.7, 29.8; MS (ESI) m/z 531.2 ([M þ H]þ calcd for C27H30F3N4Oþ4
3
2
2
99.5 (d, 2C JCF ¼ 28.8 Hz), 97.4 (t, JCF ¼ 26.3 Hz), 78.5, 70.2, 70.2,
69.1, 66.1, 57.7, 36.5; MS (ESI) m/z 416.1 ([M þ Na]þ calcd for
C19H21F2N3NaOþ4 416.1)
531.2)
5.1.3.17. (2S,3S,5R)-2-azido-1-(3,5-difluoro-phenoxy)-,6-(phenylamino)-
5-methoxy-hexan-3-ol (12a). (128 mg, 0.25 mmol) of 11a was stir-
red in a solution of TFA (10%) in freshly distilled DCM (1 mL) at r t
for 40 min. The reaction was evaporated then co-evaporated 5
times with toluene before purification by silica column chroma-
5.1.3.14. N-(4-fluoro-phenyl)-2-nitro-benzenesulfonamide.. Nosylchlo-
ride (177 mg, 0.8 mmol) and pyridine (194
mL, 2.4 mmol) were added
to a solution of 4-flouranilin (154 L,1.6 mmol) in freshly distilled DCM
m
(2 mL). The reaction was stirred at r t for 2 h then quenched with 10%
HCl (aq), the pH was set to w1 before extracting with DCM (2ꢁ). The
combined organic phases were washed with brine, dried over MgSO4,
concentrated and purified by silica column chromatography (toluene–
ethyl acetate, 50:1) to give N-(4-fluoro-phenyl)-2-nitro-benzenesulfo-
namide in 70% yield (165 mg, 0.56 mmol). 1H NMR (400 MHz, CDCl3)
tography (toluene–ethyl acetate, 30:1–5:1) to afford 12a in 84%
20
yield (82 mg, 0.21 mmol).
(400 MHz, CDCl3):
[
a
]
þ2.4 (c 1, CHCl3); 1H NMR
D
d
7.30–7.24 (m, 2H), 7.00–6.88 (m, 3H), 6.49–
6.41 (m, 3H), 4.26 (dd, 1H, J ¼ 3.9, 10.3 Hz), 4.15 (dd 1H. J ¼ 8.2,
10.3 Hz), 4.06–4.01 (m, 1H), 3.84–3.77 (m, 1H), 3.72–3.67 (m, 1H),
3.50–3.38 (m, 1H), 3.41 (s, 3H), 3.32–3.24 (m, 1H), 2.07–1.95 (m,
d
7.86 (dd, 1H, J ¼ 1.3, 8.0 Hz), 7.76 (dd, 1H, J ¼ 1.4, 7.8 Hz), 7.71 (dt, 1H,
1H), 1.87–1.77 (m, 1H); 13C NMR (100.5 MHz, CDCl3):
d 165.3 (d,
1
3
J ¼ 1.4, 7.8 Hz), 7.58 (dt, 1H, J ¼ 1.3, 7.7 Hz) 7.16 (m, 2H), 6.96 (m, 2H);
1JCF ¼ 245.3 Hz), 165.1 (d, JCF ¼ 245.3 Hz), 160.2 (t, JCF ¼ 13.6 Hz),
144.1, 129.8, 116.5, 98.6 (d, 2C, 2JCF ¼ 28.8 Hz), 97.2 (t,
2JCF ¼ 25.8 Hz), 68.9, 68.8, 64.6, 57.0, 48.8, 35.6; MS (ESI) m/z 393.2
([M þ H]þ calcd for C19H23F2N4Oþ3 393.2)
13C NMR (100 MHz, CDCl3)
d
161.4 (d, JCF ¼ 246.8 Hz), 148.4, 134.2,
1
4
3
132.7, 132.0, 131.9, 131.4 (d, JCF ¼ 3.4 Hz), 126.1 (d, JCF ¼ 8.6 Hz 2C),
125.4, 116.5 (d, 2JCF ¼ 22.9 Hz, 2C); MS (ESI) m/z 319.0 ([M þ Na]þ calcd
for C12H9FN2NaO4Sþ 319.0). 2-Nitro-N-phenyl-benzenesulfonamide and
N-(benzyl)-2-nitro-benzenesulfonamide were prepared as described
above and in accordance with previously published data [32,33].
5.1.3.18. (2S,3S,5R)-2-azido-1-(3,5-difluoro-phenoxy)-,6-(4-fluoro-
phenylamino)-5-methoxy-hexan-3-ol (12b). Title compound 12b
was obtained from 11b, as described for 12 a, in 99% yield (38 mg,
20
20
5.1.3.15. [(2R,4S,5S)-5-azido-6-(3,5-difluoro-phenoxy)-2-methoxy-4-
93
m
mol). [
a
]
ꢂ12.1 (c 1.0, MeOH); [
a
]
ꢂ12.1 (c 1.0, MeOH);1H
D
D
(4-methoxy-benzyloxy)-hexyl]-(phenyl)-amine (11a). DIAD (105
mL,
NMR (400 MHz, CD3OD) d 6.85 (m, 2H), 6.65 (m, 2H), 6.55 (m,
0.53 mmol, 1.4 equiv) was added drop wise at 0 ꢀC to a stirred
solution of 8 (167 mg, 0.38 mmol, 1 equiv), 2-nitro-N-phenyl-ben-
zenesulfonamide (144 mg, 0.52 mmol,1.36 equiv) and Ph3P (140 mg,
0.53 mmol, 1.4 equiv) in freshly distilled DCM (5 mL). The reaction
mixture was allowed to reach r t, then left stirring over night. After
evaporation the crude product was used without further purifica-
tion. The crude product was dissolved in MeCN (5 mL) and thio-
3H), 4.26 (dd, 1H, J ¼ 3.9, 10.1 Hz), 4.15 (dd, 1H, J ¼ 7.9, 10.1 Hz),
3.95 (m, 1H), 3.75 (m, 1H), 3.55 (m, 1H), 3.40 (s, 3H), 3.27 (d, 1H,
J ¼ 4.3, 13.2 Hz), 3.14 (d, 1H, J ¼ 6.1, 13.2 Hz), 1.88 (m, 2H); 13C NMR
1
(100 MHz, CD3OD)
d
165.2 (d, JCF ¼ 245.4 Hz), 165.1 (d,
1JCF ¼ 245.4 Hz), 161.0 (t, 3JCF ¼ 13.8 Hz), 157.2 (d, 1JCF ¼ 233.4 Hz),
146.6 (d, 4JCF ¼ 1.5 Hz), 129.6, 116.3 (d, 2C, 2JCF ¼ 22.3 Hz), 115.1 (d,
2C, 3JCF ¼ 7.4 Hz), 99.5 (d, 2C, 2JCF ¼ 28.9 Hz), 97.4 (d,
2JCF ¼ 26.7 Hz), 78.5, 70.2, 69.1, 66.3, 57.2, 48.0, 36.7; MS (ESI) m/z
411.2 ([M þ H]þ calcd for C19H22F3N4Oþ3 411.2)
phenol (118
mL, 1.15 mmol, 3 equiv) was added followed by
potassium carbonate (158 mg, 1.15 mmol, 3 equiv). The reaction
was heated to 50 ꢀC and followed by TLC. After 3 h all starting
material was consumed. The reaction mixture was evaporated, co-
evaporated with toluene and then filtered through a short silica
plug using ethyl acetate as eluent before concentration and puri-
5.1.3.19. N-[5S-azido-6-(3,5-difluoro-phenoxy)- 2R-methoxy-4S-(4-
methoxy-benzyloxy)-hexyl]-N-benzyl-2-nitro-benzenesulfonamide
(13). DIAD (66
m
L, 0.34 mmol) was added drop wise at 0 ꢀC to
fication by silica column chromatography (toluene–ethyl acetate,
a stirred solution of 8 (105 mg 0.24 mmol), N-(benzyl)-2-nitro-
benzenesulfonamide (95 mg, 0.33 mmol) and Ph3P (88 mg,
0.34 mmol) in freshly distilled DCM (5 mL). The reaction mixture
was allowed to reach r t, then left stirring over night. After evap-
oration the crude product was purified by silica column chroma-
20
50:1) to give 11a in 74% yield (145 mg, 0.28 mmol). [
a
]
þ2.6 (c 1,
D
CHCl3); 1H NMR (400 MHz, CDCl3):
d
7.27–7.16 (m, 4H), 6.89–6.85
(m, 2H), 6.77–6.71 (m, 1H), 6.64–6.59 (m, 2H), 6.48–6.35 (m, 3H),
4.56 (d, 1H, J ¼ 11.4 Hz), 4.47 (d, 1H, J ¼ 11.4 Hz), 4.06–4.01 (m, 2H),
3.92–3.84 (m, 2H), 3.80–3.75 (m, 1H), 3.79 (s, 3H), 3.63–3.56 (m,
1H), 3.39 (s, 3H), 3.30–3.22 (m, 1H), 3.16–3.07 (m,1H), 1.98–1.92 (m,
tography (toluene–ethyl acetate, 50:1) to give 13 in 61% yield
20
(104 mg, 0.146 mmol). [
a
]
ꢂ1.8 (c 1, CHCl3); 1H NMR (400 MHz,
D
2H); 13C NMR (100.5 MHz, CDCl3);
d
163.8 (d, 1JCF ¼ 246.9 Hz), 163.7
CD3OD): d 8.01 (m, 1H), 7.78 (m, 2H), 7.71 (m, 1H), 7.30–7.18 (m, 7H),
(d, 1JCF ¼ 246.9 Hz), 160.1 (t, 3JCF ¼ 13.8 Hz), 159.7, 148.3, 130.1, 129.7,
6.84 (m, 2H), 6.56–6.44 (m, 3H), 4.64 (m, 2H), 4.41 (m, 2H), 4.05 (m,
1H), 3.98 (m, 1H), 3.76 (s, 3H), 3.73 (m, 1H), 3.67 (m, 1H), 3.45–3.35
2
129.5, 117.9, 114.1, 113.2, 98.6 (d, 2C, JCF ¼ 28.6 Hz), 97.1 (t,
2JCF ¼ 25.8 Hz), 76.3, 74.6, 71.9, 68.5, 62.4, 56.9, 55.4, 45.9, 32.8; MS
(ESI) m/z 513.2 ([M þ H]þ calcd for C27H31F2N4Oþ4 513.2)
(m, 3H), 3.10 (s, 3H), 1.81–1.72 (m, 1H), 1.65–1.56 (m, 1H); 13C NMR
1
(100.5 MHz, CD3OD):
d
163.9 (d, JCF ¼ 247.0 Hz), 163.7 (d,
1JCF ¼ 247.0 Hz), 161.8 (t, JCF ¼ 13.6 Hz), 161.0, 149.4, 137.3, 135.2,
3
5.1.3.16. [(2R,4S,5S)-5-azido-6-(3,5-difluoro-phenoxy)-2-methoxy-4-
(4-methoxy-benzyloxy)-hexyl]-(4-fluoro-phenyl)-amine (11b). Title
compound 11b was obtained from 8 and N-(4-fluoro-phenyl)-2-
nitro-benzenesulfonamide, as described for 11a, in 50% yield
134.6, 133.0, 131.9, 131.3, 131.0, 129.7, 129.5, 129.0, 125.4, 114.8, 99.5
2
2
(d,2C, JCF ¼ 28.8 Hz), 97.4 (t, JCF ¼ 26.5 Hz), 78.0, 75.7, 72.9, 69.4,
63.8, 57.3, 55.7, 53.6, 51.6, 33.4; MS (ESI) m/z 734.2 ([M þ Na]þ calcd
for C34H35F2N5NaO8Sþ 734.2).

J. Adrian Meredith et al. / European Journal of Medicinal Chemistry 45 (2010) 542–554
551
5.1.3.20. N-[5S-azido-6-(3,5-difluoro-phenoxy)-4S-hydroxy-2R-meth-
oxy-hexyl]-N-benzyl-2-nitro-benzenesulfonamide (14). DDQ
2.97 (s, 3H),1.78 (m, 2H),1.64 (d, 3H, J ¼ 7.0 Hz); 13C NMR (100 MHz,
1
CD3OD)
d
169.1, 167.8, 165.3 (d, JCF ¼ 245.0 Hz), 165.1 (d,
3
(56 mg, 0.25 mmol) was added at 0 ꢀC to a stirred solution of 13
(136 mg, 0.19 mmol) in DCM:H20 (19:1, 10 mL). The reaction was
allowed to reach r t then stirred for an additional 3 h. The reaction
mixture was washed with saturated NaHCO3 (aq) and water and
dried over Na2SO4 before it was concentrated and purified by silica
1JCF ¼ 245.0 Hz), 162.3 (t, JCF ¼ 13.9 Hz), 145.1, 143.8, 137.5, 137.1,
129.6, 129.4, 129.3, 129.2, 128.2, 127.3, 125.9, 98.6 (d, 2C,
2
2JCF ¼ 29.1 Hz), 97.2 (t, JCF ¼ 26.4 Hz), 80.8, 68.8, 68.2, 63.6, 57.4,
54.2, 51.0, 38.3, 36.3, 35.9, 22.1; HRMS (ESI) m/z 650.2340 ([M þ H]þ
calcd for C31H38F2N3O8Sþ 650.2342).
column chromatography (toluene–ethyl acetate, gradient 60:1–
20
10:1) to give 14 in 64% yield (72 mg, 0.12 mmol). [
a
]
þ9.6 (c 1.0,
5.1.3.23. N-[(1S,2S,4R)-1-(3,5-difluoro-phenoxymethyl)-2-hydroxy-4,5-
dimethoxy-pentyl]-5-(methanesulfonyl-methyl-amino)-N0-(1R-phenyl-
ethyl)-isophthalamide (2b). Title compound 2b was obtained from
D
MeOH); 1H NMR (400 MHz, CDCl3)
d
8.01 (m, 1H), 7.69 (m, 3H), 7.27
(m, 5H), 6.47 (m, 3H), 4.64 (d, 2H, J ¼ 7.9 Hz), 4.19 (dd, 1H, J ¼ 4.2,
9.8 Hz), 4.07 (dd, 1H, J ¼ 8.0, 9.7 Hz), 3.83 (m, 1H), 3.59 (m, 1H),
3.56–3.36 (m, 3H), 3.23 (s, 3H), 1.63 (m, 2H); 13C NMR (100 MHz,
5b as described for 2a, in 43% yield (19 mg, 29 mmol). 2b was further
20
purified by LC-MS. [
CD3OD)
a]
ꢂ20.3 (c 1.0, MeOH); 1H NMR (400 MHz,
D
CDCl3)
d
163.9 (d, 1JCF ¼ 246.8 Hz),163.7 (d, 1JCF ¼ 246.8 Hz),160.2 (t,
d
8.24 (t, 1H, J ¼ 1.6 Hz), 8.04 (m, 2H), 7.43–7.18 (m, 5H),
3JCF ¼ 13.4 Hz), 148.8, 135.8, 133.8, 133.5, 131.8, 131.4, 128.9, 128.5,
128.3, 124,3, 98.6 (d, 2C, 2JCF ¼ 28.6 Hz), 97.1 (t, 2JCF ¼ 25.8 Hz), 79.7,
69.8, 68.8, 64.4, 57.5, 53.1, 49.8, 35.4; MS (ESI) m/z 614.2 ([M þ Na]þ
calcd for C26H27F2N5NaO7Sþ 614.2).
6.62 (m, 2H), 6.51(m, 1H), 5.25 (dd, 1H, J ¼ 7.0, 14.0 Hz), 4.54 (m,
1H), 4.26 (dd,1H, J ¼ 6.5, 9.8 Hz), 4.16 (m, 2H), 3.60 (m 1H), 3.50 (dd,
1H, J ¼ 3.6, 10.5 Hz), 3.43 (dd, 1H, J ¼ 5.2, 10.5 Hz). 3.39 (s, 3H), 3.37
(s, 3H), 3.33 (s, 3H), 2.97 (s, 3H), 1.77 (m, 2H), 1.58 (d, 3H, J ¼ 7.1 Hz);
13C NMR (100 MHz, CD3OD)
d
169.0, 167.8, 165.3 (d, 1JCF ¼ 245.0 Hz),
5.1.3.21. N-[5S-azido-6-(3,5-difluoro-phenoxy)-4R-hydroxy-2R-metho-
165.1 (d, 1JCF ¼ 245.0 Hz), 162.3 (t, 3JCF ¼ 13.6 Hz), 145.1, 143.8, 137.5,
xy-hexyl]-N-benzyl-2-nitro-benzenesulfonamide
(15). 14
(73 mg,
137.1, 129.6, 129.3, 129.3, 129.2, 128.2, 127.3, 125.9, 99.6 (d, 2C,
2
0.12 mmol), p-nitro benzaldehyde (37 mg, 0.22 mmol) and Ph3P
(55 mg, 0.21 mmol) was solved in THF (3 mL) and cooled to 0 ꢀC
before DIAD (42 mL, 0.21 mmol) was added drop wise. The reaction
2JCF ¼ 28.7 Hz), 97.2 (t, JCF ¼ 26.4 Hz), 79.1, 75.0, 68.8, 68.2, 59.4,
57.6, 54.2, 51.0, 38.3, 36.6, 35.9, 22.1; HRMS (ESI) m/z 686.2302
([M þ Na]þ calcd for C32H39F2N3NaO8Sþ 686.2318).
mixture was allowed to reach r t, then left stirring over night. After
concentration and purification on a short silica plug the crude
product was solved in 0.1 M NaOMe (10 mL), stirred in r t for 30 min,
neutralized with Dowex Hþ, filtered, concentrated and purified by
5.1.3.24. N-[(1S,2S,4R)-1-(3,5-difluoro-phenoxymethyl)-2-hydroxy-
5-isobutoxy-4-methoxy-pentyl]-5-(methanesulfonyl-methyl-amino)-
N0-(1R-phenyl-ethyl)-isophthalamide (2c). Title compound 2c was
silica column chromatography (toluene–ethyl acetate 10:1) to yield
obtained from 5c as described for 2a, in 85% yield (29 mg, 41 mmol).
20
20
the product 15 in 51% yield (37 mg, 62
m
mol). [
a
]
þ2.6 (c 1, CHCl3);
2c was further purified by LC-MS. [
a
]
ꢂ15.7 (c 1.0, MeOH); 1H NMR
D
D
1H NMR (500 MHz, CDCl3):
d
8.00 (dd, 1H, J ¼ 1.3, 7.9 Hz), 7.72–7.61
(400 MHz, CD3OD)
d
8.25 (t, 1H, J ¼ 1.6 Hz), 8.05 (m, 2H), 7.40 (m,
(m, 3H), 7.32–7.20 (m, 5H), 6.45 (m, 3H), 4.67 (d, 1H, J ¼ 15.5 Hz), 4.56
(d, 1H, J ¼ 15.5 Hz), 4.20 (dd, 1H, J ¼ 3.3, 9.9 Hz), 3.99 (dd, 1H, J ¼ 7.9,
9.9 Hz), 3.86 (m, 1H), 3.66 (m, 1H), 3.61 (m, 1H), 3.43 (d, 1H,
J ¼ 5.8 Hz), 3.21 (s, 3H), 2.87 (d, 1H, J ¼ 4.7 Hz), 1.70 (m, 1H), 1.61 (m,
2H), 7.33 (m, 2H), 7.23 (m, 1H), 6.62 (m, 2H), 6.51 (m, 1H), 5.25 (dd,
1H, J ¼ 7.1,14.1 Hz), 4.55 (m,1H), 4.26 (dd,1H, J ¼ 6.6, 9.8 Hz), 4.17 (m,
2H), 3.60 (m,1H), 3.49 (m, 2H). 3.40 (s, 3H), 3.37 (s, 3H), 3.19 (m, 2H),
2.97 (s, 3H), 1.79 (m, 3H), 1.58 (d, 3H, J ¼ 7.1 Hz), 0.85 (d, 6H,
1H), 1.58 (s, 1H); 13C NMR (125.8 MHz, CDCl3):
d
163.8 (d,
J ¼ 6.7 Hz); 13C NMR (100 MHz, CD3OD)
d 169.0, 167.8, 165.3 (d,
1
3
1
3
1JCF ¼ 247.2 Hz), 163.7 (d, JCF ¼ 247.2 Hz), 160.2 (t, JCF ¼ 13.8 Hz),
148.1, 135.4, 133.8, 131.9, 131.4, 129.0, 128.5, 128.3, 124.4, 98.7 (d, 2C,
2JCF ¼ 28.7 Hz), 97.2 (t, 2JCF ¼ 25.7 Hz), 78.0, 68.8, 68.3, 65.0, 58.1, 52.8,
49.7, 34.0; MS (ESI) m/z 614.2 ([M þ Na]þ calcd for C26H27F2N5NaO7Sþ
614.2).
1JCF ¼ 245.0 Hz), 165.1 (d, JCF ¼ 245.0 Hz), 162.3 (t, JCF ¼ 13.9 Hz),
145.0, 143.8, 137.4, 137.0. 129.6, 129.3, 128.2, 127.3, 125.9, 99.6 (d, 2C,
2JCF ¼ 28.8 Hz), 97.2 (t, JCF ¼ 26.3 Hz), 79.4, 79.2, 73.4, 68.7, 68.2,
2
57.7, 54.1, 51.0, 38.3, 36.9, 35.9, 29.6, 19.7, 19.7; HRMS (ESI) m/z
728.2790 ([M þ Na]þ calcd for C35H45F2N3NaO8Sþ 728.2778).
5.1.3.22. N-[(1S,2S,4R)-1-(3,5-difluoro-phenoxymethyl)-2,5-dihydroxy-
4-methoxy-pentyl]-5-(methanesulfonyl-methyl-amino)-N0-(1R-phenyl-
ethyl)-isophthalamide (2a). Palladium on charcoal (w5%) was added
5.1.3.25. N-[(1S,2S,4R)-5-cyclopropylmethoxy-1-(3,5-difluoro-phe-
noxymethyl)-2-hydroxy-4-methoxy-pentyl]-5-(methanesulfonyl-
(2d). Title
compound 2d was obtained from 5d as described for 2a, in 65%
methyl-amino)-N0-(1R-phenyl-ethyl)-isophthalamide
to a solution of 3 (19 mg, 59 mmol, 1 equiv) in MeOH (3 mL) under
20
argon atmosphere. The flask was evacuated followed by addition of
hydrogen gas (1 atm). The reaction mixture was stirredfor 3 h before
the Pd/C wasfiltered off and the filtratewas concentratedtoyield the
crude product that was used in the next step without further puri-
yield (30 mg, 42 mmol). 2d was further purified by LC-MS. [a]
D
ꢂ17.7 (c 1.0, MeOH); 1H NMR (400 MHz, CD3OD)
d 8.25 (t, 1H,
J ¼ 1.5 Hz), 8.04 (m, 2H), 7.44–7.08 (m, 5H), 6.62 (m, 2H), 6.51 (m,
1H), 5.25 (dd, 1H, J ¼ 7.0, 14.0 Hz), 4.55 (m, 1H), 4.27 (dd, 1H, J ¼ 6.5,
9.9 Hz), 4.17 (m, 2H), 3.58 (m, 2H), 3.50 (m, 1H). 3.40 (s, 3H), 3.37 (s,
3H), 3.28 (m, 2H), 2.97 (s, 3H), 1.79 (t, 2H, J ¼ 6.5 Hz), 1.58 (d, 3H,
J ¼ 7.1 Hz), 0.97 (m, 1H), 0.46 (m, 2H), 0.16 (m, 2H); 13C NMR
fication. Carboxylic acid A (22 mg, 59
dry DCM (2 mL) and PyBOP (31 mg, 59
followed by DIPEA (11 L, 59 mol, 1 equiv). After 30 min the crude
amine (1 equiv dissolved in DCM) was added to the mixture, fol-
lowed by DIPEA (11 L, 59 mol, 1 equiv). After 2 h, the reaction
mmol,1 equiv) was dissolved in
mmol, 1 equiv) was added
m
m
(100 MHz, CD3OD)
d
169.0, 167.8, 165.3 (d, 1JCF ¼ 245.0 Hz), 165.1 (d,
3
m
m
1JCF ¼ 245.0 Hz), 162.3 (t, JCF ¼ 14.0 Hz), 145.0, 143.8, 137.5, 137.0,
mixture was diluted with DCM, washed with Na2CO3 (sat.) and
NH4Cl (aq). The aqueous layers were extracted with DCM (3x), and
the combined organic phases were washed with brine, dried
(MgSO4), concentrated and purified by silica column chromatog-
raphy (toluene–ethyl acetate gradient 10:1–1:6) to yield compound
129.9, 129.6, 129.3, 129.3, 128.2, 127.3, 125.9, 99.6 (d, 2C,
2JCF ¼ 29.0 Hz), 97.2 (t, JCF ¼ 26.3 Hz), 79.2, 77.0, 72.7, 68.7, 68.2,
2
57.6, 54.2, 51.0, 38.3, 36.7, 35.9, 22.1, 11.4, 3.4, 3.4; HRMS (ESI) m/z
726.2647 ([M þ Na]þ calcd for C35H43F2N3O8SNaþ 726.2631).
2a in 39% yield (15 mg, 23
m
mol). 2a was further purified by LC-MS.
5.1.3.26. N-[(1S,2S,4R)-1-(3,5-difluoro-phenoxymethyl)-2-hydroxy-
4-methoxy-5-(4,4,4-trifluoro-butoxy)-pentyl]-5-(methanesulfonyl-
20
[
a]
ꢂ23.4 (c 0.53, MeOH); 1H NMR (400 MHz, CD3OD)
d 8.24 (t,1H,
D
J ¼ 1.6 Hz), 8.04 (m, 2H), 7.44–7.08 (m, 5H), 6.65–6.47 (m, 3H), 5.25
(m, 1H), 4.55 (m, 1H), 4.27 (dd, 1H, J ¼ 6.5, 9.9 Hz), 4.17 (m, 2H), 3.69
(dd, 1H, J ¼ 3.4, 11.4 Hz), 3.54–3.44 (m, 2H). 3.39 (s, 3H), 3.38 (s, 3H),
methyl-amino)-N0-(1R-phenyl-ethyl)-isophthalamide
(2e). Title
compound 2e was obtained from 5e as described for 2a, in 39%
20
yield (17 mg, 22 mmol). 2e was further purified by LC-MS. [a]
D

552
J. Adrian Meredith et al. / European Journal of Medicinal Chemistry 45 (2010) 542–554
ꢂ18.4 (c 0.75, MeOH); 1H NMR (400 MHz, CD3OD)
d
8.25 (t, 1H,
(toluene–ethyl acetate gradient 10:1–1:6) to yield 2h in 82% yield
20
J ¼ 1.6 Hz), 8.04 (m, 2H), 7.42–7.20 (m, 5H), 6.62 (m, 2H), 6.51 (m,
1H), 5.25 (dd, 1H, J ¼ 7.0, 14.0), 4.55 (m, 1H), 4.26 (dd, 1H, J ¼ 6.5,
9.89 Hz), 4.17 (m, 2H), 3.65–3.45 (m, 5H). 3.40 (s, 3H), 3.37 (s, 3H),
(30 mg, 43
m
mol). 2h was further purified by LC-MS. [
a
]
ꢂ23.0 (c
D
0.80, MeOH); 1H NMR (400 MHz, CD3OD)
d
8.24 (t, 1H, J ¼ 1.6 Hz),
8.04 (m, 2H), 7.43–7.21 (m, 5H), 6.65–6.48 (m, 3H), 5.86 (m, 1H), 5.24
(m, 2H), 5.11 (m, 1H), 4.55 (m, 1H), 4.26 (dd, 1H, J ¼ 6.5, 9.9 Hz), 4.17
(m, 2H), 3.98 (m, 2H), 3.61 (m, 1H), 3.56 (dd, 1H, J ¼ 3.7,10.5 Hz), 3.48
(dd, 1H, J ¼ 5.1, 10.5 Hz), 3.40 (s, 3H), 3.37 (s, 3H), 2.97 (s, 3H), 1.79 (t,
2H, J ¼ 6.6 Hz), 1.58 (d, 3H, J ¼ 7.1 Hz); 13C NMR (100 MHz, CD3OD)
2.97 (s, 3H), 2.18 (m, 2H), 1.77 (m, 4H), 1.59 (d, 3H, J ¼ 7.1 Hz); 13C
1
NMR (100 MHz, CD3OD)
d
169.0, 167.8, 165.3 (d, JCF ¼ 245.2 Hz),
165.1 (d, 1JCF ¼ 245.2 Hz), 162.3 (t, 3JCF ¼ 13.8 Hz), 145.1, 143.8, 137.5,
137.0, 129.6, 129.4, 129.3, 128.2, 127.3, 125.9, 99.6 (d, 2C,
2JCF ¼ ,28.9 Hz), 97.2 (t, JCF ¼ 26.4 Hz), 79.1, 73.2, 70.6, 68.8, 68.2,
d
169.0, 167.8, 165.3 (d, JCF ¼ 244.6 Hz), 165.1 (d, JCF ¼ 244.6 Hz),
3
2
1
1
2
57.7, 54.1, 51.0, 38.3, 36.7, 35.9, 31.4 (dd, JCF ¼ 28.7, 86.5 Hz), 23.5
162.3 (t, JCF ¼ 13.8 Hz), 145.0, 143.8, 137.5, 137.0, 136.0, 129.6, 129.3,
(dd, JCF ¼ 3.1, 9.1 Hz), 22.1; HRMS (ESI) m/z 760.2677 ([M þ H]þ
128.2, 127.2, 125.9, 117.2, 99.6 (d, 2C, JCF ¼ 28.8 Hz), 97.2 (t,
3
2
calcd for C35H43F5N3O8Sþ 760.2686).
2JCF ¼ 26.4 Hz), 79.2, 73.2, 72.4, 68.8, 68.2, 57.7, 54.1, 51.0, 38.3, 36.7,
35.9, 22.1; HRMS (ESI) m/z 712.2496 ([M þ Na]þ calcd for
C34H41F2N3O8SNaþ 712.2475).
5.1.3.27. N-[(1S,2S,4R)-1-(3,5-difluoro-phenoxymethyl)-2-hydroxy-
4-methoxy-5-phenoxy-pentyl]-5-(methanesulfonyl-methyl-amino)-
N0-(1R-phenyl-ethyl)-isophthalamide (2f). Title compound 2f was
5.1.3.30. N-[(1S,2S,4R)-1-(3,5-difluoro-phenoxymethyl)-5-(4-fluoro-
benzyloxy)-2-hydroxy-4-methoxy-pentyl]-5-(methanesulfonyl-methyl-
amino)-N0-(1R-phenyl-ethyl)-isophthalamide (2i). Title compound 2i
obtained from 10 as described for 2a, in 61% yield (27 mg,
20
37
m
mol). 2f was further purified by LC-MS. [
a
]
ꢂ18.7 (c 0.3,
D
MeOH); 1H NMR (400 MHz, CDCl3)
d
8.15 (m, 1H), 7.98 (m, 2H),
was obtained from 5g as described for 2h, in 70% yield (30 mg,
20
7.37 (m, 4H), 7.28 (m, 2H), 6.95 (m, 2H), 6.87 (m, 2H), 6.60 (d, 1H,
J ¼ 7.6 Hz), 6.51 (m, 2H), 6.42 (m, 1H), 5.32 (m, 1H), 4.47 (m, 1H),
4.37 (m, 1H), 4.15 (m, 1H), 4.08 (dd, 1H, J ¼ 5.2, 9.3 Hz), 4.03 (dd,
1H, J ¼ 4.6, 9.8 Hz), 3.94 (m, 2H), 3.55 (s, 3H), 3.37 (s, 3H), 2.86 (s,
39
m
mol). 2i was further purified by LC-MS. [
a]
ꢂ1.9 (c 1.0,
D
MeOH);1H NMR (400 MHz, CD3OD)
d
8.24 (t, 1H, J ¼ 1.5 Hz), 8.04
(m, 2H), 7.40 (m, 2H), 7.30 (m, 4H), 7.22 (m, 2H), 7.12 (m, 1H), 6.98
(m, 2H), 6.60 (m, 2H), 6.50 (m, 2H), 5.24 (dd, 1H, J ¼ 7.0, 14.1), 4.54
(s,1H), 4.47 (d, 2H, J ¼ 4.5 Hz), 4.25 (dd,1H, J ¼ 6.6,10.0 Hz), 4.17 (m,
2H), 3.64 (m, 1H), 3.58 (dd, 1H, J ¼ 3.7, 10.4 Hz), 3.51 (dd, 1H, J ¼ 5.0,
10.4 Hz), 3.38 (s, 3H), 3.35 (s, 3H), 2.94 (s, 3H), 1.81 (m, 2H), 1.58 (d,
3H), 1.89 (m, 2H), 1.63 (d, 3H, J ¼ 7.0 Hz); 13C NMR (125 MHz,
1
CDCl3)
d
165.8, 163.9 (d, JCF ¼ 246.0 Hz), 164.6, 163.8 (d,
1JCF ¼ 246.0 Hz), 160.4 (t, 3JCF ¼ 13.5 Hz), 158.4, 142.8, 142.5, 136.2,
135.5, 129.7, 129.0, 127.9, 127.8, 127.6, 126.4, 124.1, 121.4, 114.6. 98.7
(d, 2C, 2JCF ¼ 28.8 Hz), 96.9 (t, 2JCF ¼ 25.9 Hz), 80.3, 69.2, 69.1, 67.0,
58.1, 53.1, 50.0, 38.1, 36.3, 35.6, 21.8; HRMS (ESI) m/z 726.2627
([M þ H]þ calcd for C37H42F2N3O8Sþ 726.2655).
3H, J ¼ 7.0 Hz); 13C NMR (100 MHz, CD3OD)
d 169.0, 167.7, 165.2 (d,
1JCF ¼ 245.1 Hz), 165.1, (d, 1JCF ¼ 245.1 Hz), 163.7 (d, 1JCF ¼ 244.4 Hz),
3
162.3 (t, JCF ¼ 13.8 Hz), 145.0, 143.8, 137.4, 137.0, 135.6 (d,
4JCF ¼ 3.3 Hz), 130.7 (d, 2C, JCF ¼ 8.2 Hz), 129.9, 129.6, 129.4, 129.3,
3
129.2, 128.2, 127.3, 126.3, 125.9, 116.0 (d, 2C, 2JCF ¼ 21.8 Hz), 99.6 (d,
2C, 2JCF ¼ 28.7 Hz), 97.2 (t, 2JCF ¼ 26.3 Hz), 79.1, 73.4, 72.3, 68.8, 68.2,
57.7, 54.1, 60.0, 38.3, 36.7, 35.9, 22.1; HRMS (ESI) m/z 758.2683
([M þ H]þ calcd for C38H43F3N3O8Sþ 758.2717).
5.1.3.28. N-[1S-(3,5-difluoro-phenoxymethyl)-2S-hydroxy-4R-methoxy-
5-phenylamino-pentyl]-5-(methanesulfonyl-methyl-amino)-N0-(1R-
phenyl-ethyl)-isophthalamide (2g). Title compound 2g was
obtained from 12a as described for 2a, in 57% yield (34 mg,
20
47
m
mol). 2g was further purified by LC-MS. [
a
]
þ17.4 (c 1, CHCl3);
5.1.3.31. N-[(1S,2S,4R)-1-(3,5-difluoro-phenoxymethyl)-2-hydroxy-
4-methoxy-5-(3-methoxy-benzyloxy)-pentyl]-5-(methanesulfonyl-
D
1H NMR (400 MHz, CD3OD):
d
8.13 (m, 1H), 7.96 (m, 2H), 7.36 (m,
4H), 7.28 (m, 2H), 7.17 (m, 2H), 7.0 (d, 1H, J ¼ 8.7 Hz), 6.74 (m, 2H),
6.64 (m, 2H), 6.49 (m, 2H), 6.41 (m, 1H), 5.30 (m, 1H), 4.44 (m, 1H),
4.31 (m, 1H), 4.13 (m, 1H), 4.07 (dd, 1H, J ¼ 5.5, 9.4 Hz), 3.82 (m, 1H),
3.44 (s, 3H), 3.33 (s, 3H), 3.19 (dd, 1H, J ¼ 3.1, 12.8 Hz) 2.83 (s, 3H),
1.97 (m, 1H), 1.77 (m, 1H), 1.60 (d, 3H, J ¼ 7.0 Hz); 13C NMR
methyl-amino)-N0-(1R-phenyl-ethyl)-isophthalamide
(2j). Title
compound 2j was obtained from 5h as described for 2h, in 92% yield
20
(53 mg, 69
m
mol). 2j was further purified by LC-MS. [
a]
ꢂ12.4 (c
D
1.0, MeOH); 1H NMR (400 MHz, CD3OD)
d
8.24 (t,1H, J ¼ 1.5 Hz), 8.04
(m, 21H), 7.40 (m, 2H), 7.32 (m, 2H), 7.26–7.09 (m, 2H), 6.85 (m, 2H),
6.78 (m,1H), 6.60 (m, 2H), 6.50 (m,1H), 5.24 (dd,1H, J ¼ 7.0,14.0 Hz),
4.59 (bs, 1H), 4.55 (m, 1H), 4.48 (d, 2H, J ¼ 5.4 Hz), 4.24 (dd, 1H,
J ¼ 6.5, 9.9 Hz), 4.15 (m, 2H), 3.73 (s, 3H), 3.67–3.56 (m, 2H), 3.51 (dd,
1H, J ¼ 4.8, 10.5 Hz), 3.39 (s, 3H), 3.35 (s, 3H), 2.94 (s, 3H), 1.81 (m,
(100.5 MHz, CD3OD):
d
169.0, 167.7, 163.8 (d, 1JCF ¼ 246.2 Hz), 163.7
(d, 1JCF ¼ 246.2 Hz), 162.2 (t, JCF ¼ 13.9 Hz), 150.0, 145.0, 143.7, 138.9,
137.4, 137.0, 130.1, 129.9, 129.6, 129.3, 129.2, 128.2, 127.3, 126.3,
126.0, 118.2, 114.1, 99.6 (d, 2C, 2JCF ¼ 28.7 Hz), 97.2 (t, 2JCF ¼ 26.4 Hz),
78.8, 68.8, 68.3, 57.4, 54.3, 50.9, 47.3, 38.3, 37.5, 35.9, 22.1; HRMS
(ESI) m/z 725.2767 ([M þ H]þ calcd for C37H43F2N4O7Sþ 725.2815).
2H), 1.58 (d, 3H, J ¼ 7.1 Hz); 13C NMR (100 MHz, CD3OD)
d 169.0,
1
1
167.7, 166.3, 165.2 (d, JCF ¼ 245.4 Hz), 165.1, (d, JCF ¼ 245.4 Hz),
162.3 (t, 3JCF ¼ 13.9 Hz), 161.2, 145.0, 143.8, 141.1, 137.4, 137.0, 130.4,
129.6, 129.3, 129.2, 128.2, 127.3, 125.9, 120,9, 114.1, 99.6 (d, 2C,
2JCF ¼ 28.6 Hz), 97.2 (t, 2JCF ¼ 26.5 Hz), 79.1, 74.1, 72.2, 68.8, 68.2, 57.7,
55.6, 54.2, 51.0, 38.3, 36.8, 35.9, 22.1; HRMS (ESI) m/z 792.2709
([M þ Na]þ calcd for C39H45F2N3O9Sþ 792.2737).
5.1.3.29. N-[(1S,2S,4R)-5-allyloxy-1-(3,5-difluoro-phenoxymethyl)-
2-hydroxy-4-methoxy-pentyl]-5-(methanesulfonyl-methyl-amino)-N0-
(1R-phenyl-ethyl)-isophthalamide (2h). Ph3P (21 mg, 80
1.5 equiv) was added together with two drops ofwater to a solution of
the 5f (19 mg, 53 mol, 1 equiv) in MeOH (3 mL). The reaction
mmol,
m
mixture was stirred for 16 h, concentrated to yield the crude
compound which was used in the next step without further purifi-
5.1.3.32. N-[(1S,2S,4R)-5-benzyloxy-1-(3,5-difluoro-phenoxymethyl)-
2-hydroxy-4-methoxy-pentyl]-5-(methanesulfonyl-methyl-amino)-N0-
(1R-phenyl-ethyl)-isophthalamide (2k). Title compound 2k was
cation. Carboxylic acid A (20 mg, 53
dry DCM (2 mL) and PyBOP (28 mg, 53
followed by DIPEA (9 L, 53 mol, 1 equiv). After 30 min the crude
amine (1 equiv, dissolved in DCM) was added to the mixture, fol-
lowed by DIPEA (9 L, 53 mol, 1 equiv). After 2 h, the reaction
mmol, 1 equiv) was dissolved in
mmol, 1 equiv) was added
obtained from 5a as described for 2h, in 83% yield (30 mg, 40 mmol).
20
m
m
2kwasfurtherpurified by LC-MS. [
a
]
ꢂ11.8 (c 0.76, MeOH);1H NMR
D
(400 MHz, CD3OD)
d
8.24(t,1H,J ¼ 1.5 Hz), 8.04(m, 2H), 7.44–7.19(m,
m
m
10H), 6.64–6.47 (m, 3H), 5.25 (dd,1H, J ¼ 7.0,14.1), 4.54 (m, 3H), 4.25
(dd,1H, J ¼ 6.4, 9.8 Hz), 4.17 (m, 2H), 3.62(m, 2H), 3.52 (dd,1H, J ¼ 4.9,
10.4 Hz), 3.39(s, 3H), 3.36(s, 3H), 2.95 (s, 3H),1.81 (m, 2H),1.58 (d, 3H,
mixture was diluted with DCM, washed with Na2CO3 (sat.) and NH4Cl
(aq). The aqueous layers were extracted with DCM (3x), and
the combined organic phases were washed with brine, dried
(MgSO4), concentrated and purified bysilicacolumn chromatography
J ¼ 7.0 Hz); 13C NMR (100 MHz, CD3OD)
d 169.0, 167.8, 165.3 (d,
1JCF ¼ 245.1 Hz), 165.2 (d, JCF ¼ 245.1 Hz), 162.3 (t, JCF ¼ 13.9 Hz),
1
3

J. Adrian Meredith et al. / European Journal of Medicinal Chemistry 45 (2010) 542–554
553
20
145.1, 143.8,139.6,137.4, 137.0,129.6,129.4,129.3,128.8,128.7, 128.2,
127.3, 125.9, 99.6 (d, 2C, 2JCF ¼ 29.0 Hz), 97.2 (d, 2JCF ¼ 26.5 Hz), 79.1,
74.3, 72.3, 68.8, 68.2, 57.7, 54.1, 50.9, 38.3, 36.8, 35.9, 22.1; HRMS (ESI)
m/z 740.2837 ([M þ H]þ calcd for C38H44F2N3O8Sþ 740.2812).
yield (54 mg, 58
CDCl3):
m
mol). [
a
]
þ24.5 (c 1, CHCl3); 1H NMR (400 MHz,
D
d
8.08 (m, 1H), 7.98 (t, 1H, J ¼ 1.9 Hz), 7.95 (m, 1H), 7.91 (dd,
1H, J ¼ 1.3, 8.0 Hz), 7.73–7.57 (m, 3H), 7.38–7.28 (m, 4H), 7.25 (m,
4H), 7.16 (m, 2H), 6.93 (d, 1H, J ¼ 8.4), 6.73 (d, 1H, J ¼ 7.6 Hz), 6.50
(m, 2H) 6.38 (m, 1H), 5.29 (m, 1H), 4.68 (d, 1H, J ¼ 15.4 Hz), 4.46 (d,
1H, J ¼ 15.4 Hz), 4.33 (m, 2H), 4.13 (m, 2H), 3.57 (m, 2H), 3.33 (s,
3H), 3.30 (m,1H), 3.17 (s, 3H), 2.84 (s, 3H),1.75 (m, 2H),1.65 (bs,1H),
5.1.3.33. N-[(1S,2S,4R)-1-(3,5-difluoro-phenoxymethyl)-2-hydroxy-
4-methoxy-5-(4-methoxy-benzyloxy)-pentyl]-5-(methanesulfonyl-
methyl-amino)-N0-(1R-phenyl-ethyl)-isophthalamide
(2l). Title
compound 2l was obtained from 7 as described for 2h, in 39% yield
1.59 (d, 3H, J ¼ 7.0 Hz); 13C NMR (100.5 MHz, CDCl3):
d 166.1, 164.9,
1
1
163.7 (d, JCF ¼ 246.6 Hz), 163.5 (d, JCF ¼ 246.6 Hz), 160.5 (t,
3JCF ¼ 13.4 Hz), 148.0, 142.9, 142.5, 136.3, 135.7, 135.1, 134.0, 133.2,
132.0, 131.3, 129.0, 128.9, 128.5, 128.4, 128.2, 127.7, 126.4, 124.5,
20
(18 mg, 23
m
mol). 2l was further purified by LC-MS. [
a
]
ꢂ5.5 (c
D
0.76, MeOH); 1H NMR (400 MHz, CD3OD)
d
8.44 (t, 1H, J ¼ 1.5 Hz),
2
2
8.21 (m, 1H), 8.08 (m, 1H), 7.39 (m, 2H), 7.32 (m, 2H), 7.23 (m, 3H),
6.83 (m, 2H), 6.62 (m, 2H), 6.54 (m,1H), 5.21 (dd,1H, J ¼ 7.1,14.2 Hz),
4.45 (d, 2H, J ¼ 4.7 Hz), 4.28 (d, 2H, J ¼ 6.0 Hz), 3.74 (s, 3H), 3.63–3.50
(m, 3H), 3.40 (s, 3H), 3.37 (s, 3H), 3.08 (dd, 1H, J ¼ 6.0, 12.3 Hz), 2.93
(m, 4H), 2.02 (m, 1H), 1.91 (m, 1H), 1.49 (d, 3H, J ¼ 7.0 Hz); 13C NMR
124.0, 98.8 (d, 2C, JCF ¼ 28.6 Hz), 97.0 (t, JCF ¼ 25.9 Hz), 77.9, 67.8,
67.1, 58.0, 53.7, 52.8, 49.9, 48.9, 38.1, 35.8, 34.1, 21.8; HRMS (ESI) m/z
946.2591 ([M þ Na] þ calcd for C44H47F2N5NaO11Sþ2 946.2574)
5.1.3.37. N-[5-benzylamino-1S-(3,5-difluoro-phenoxymethyl)-2S-hyd-
roxy-4R-methoxy-pentyl]-5-(methanesulfonyl-methyl-amino)-N0-
(1R-phenyl-ethyl)-isophthalamide (2n). To a stirred solution of 16
(125 MHz, CD3OD)
d
179.6, 167.7, 165.2 (d, 1JCF ¼ 245.8 Hz), 165.1 (d,
1JCF ¼ 245.8 Hz), 162.1 (t, JCF ¼ 13.8 Hz), 160.8, 145.1, 143.8, 137.6,
3
135.6, 131.5, 131.1, 130.6, 129.6, 129.6, 128.1, 127.7, 127.2, 114.8, 99.6
(50 mg, 54
mmol, 1 equiv) in MeCN (3 mL) was added thiophenol
2
2
(d, 2C, JCF ¼ 28.9 Hz), 97.4 (t, JCF ¼ 26.5 Hz), 79.8, 74.0, 71.8, 68.6,
58.5, 55.7, 51.0, 41.8, 40.1, 38.2, 35.8, 31.5, 22.0;; HRMS (ESI) m/z
770.2916 ([M þ H]þ calcd for C39H46F2N3O9Sþ 770.2917).
(18 mL, 0.16 mmol, 3 equiv) and K2CO3 (22 mg, 0.16 mmol,
3 equiv). After 2 h of heating (50 ꢀC) the reaction mixture was
allowed to reach r t before it was concentrated and purified by
silica column chromatography (toluene–ethyl acetate gradient
5.1.3.34. N-[(1S,2S,4R)-1-(3,5-difluoro-phenoxymethyl)-5-(4-fluoro-
phenylamino)-2-hydroxy-4-methoxy-pentyl]-5-(methanesulfonyl-
(2m). Title
compound 2m was obtained from 12b as described for 2h, in 40%
10:1–100% ethyl acetate) to yield the compound 2n in 50% yield
20
(20 mg, 27
m
mol). 2n was further purified by LC-MS.. [
a
]
ꢂ14.8
D
methyl-amino)-N0-(1R-phenyl-ethyl)-isophthalamide
(c 1.0, CHCl3); 1H NMR (400 MHz, CDCl3)
d 8.18 (m, 1H), 8.00 (m,
2H), 7.41–7.27 (m, 10H), 7.12 (d, 1H, J ¼ 8.5 Hz), 6.75 (d, 1H,
J ¼ 7.8 Hz), 6.50 (m, 2H), 6.39 (m, 1H), 5.32 (m, 1H) 4.43 (m,
1H),4.16 (m, 2H), 4.07 (dd, 1H, J ¼ 5.6, 9.2 Hz), 3.80 (dd, 2H,
J ¼ 13.0, 33.1 Hz), 3.61 (m, 1H), 3.37 (s, 3H), 3.30 (s, 3H), 3.07 (dd,
1H, J ¼ 5.0, 12.1 Hz), 2.86 (s, 3H), 2.56 (m, 1H), 2.03 (m, 2H), 1.62
20
yield (20 mg, 27 mmol). 2m was further purified by LC-MS. [a]
D
þ14.2 (c 1.0, MeOH);1H NMR (400 MHz, CDCl3)
d 8.52 (d, 1H,
J ¼ 5.5 Hz), 8.13 (t,1H, J ¼ 1.5 Hz), 7.95 (m, 2H), 7.40–7.22 (m, 5H), 7.17
(m,1H), 6.90 (m, 2H), 6.62 (m, 2H) 6.55 (m 2H), 6.46 (m,1H) 5.31 (m,
1H), 4.81 (m, 1H), 4.48 (dd, 1H, J ¼ 3.8, 9.9 Hz), 4.41 (m, 1H), 4.18 (m,
1H), 3.95 (t, 1H, J ¼ 9.5 Hz), 3.74 (d, 1H, J ¼ 11.1 Hz), 3.54 (s, 3H), 3.33
(s, 3H), 3.29 (dd 1H, J ¼ 4.8, 11.1 Hz), 2.84 (s, 3H), 2.43 (m, 2H), 1.70
(d, 3H, J ¼ 6.7 Hz); 13C NMR (100 MHz, CDCl3)
d 165.9, 163.9 (d,
1JCF ¼ 247.2 Hz), 164.7, 163.7 (d, JCF ¼ 247.2 Hz), 160.7 (t,
3JCF ¼ 13.9 Hz), 142.9, 142.4, 138.1, 136.1, 135.8, 129.0, 128.9, 128.6,
127.9, 127.9, 127.8, 126.4, 124.1, 98.7 (d, 2C, 2JCF ¼ 28.6 Hz), 96.7 (d,
2JCF ¼ 25.9 Hz), 77.4, 67.7, 63.0, 56.8, 54.0, 53.8, 50.6, 49.9, 38.3,
38.1, 35.6, 21.8 HRMS (ESI) m/z 739.2951 ([M þ H]þ calcd for
C38H45F2N4O7Sþ 739.2972)
1
(bs, 1H), 1.60 (d, 3H, J ¼ 9.7 Hz); 13C NMR (100 MHz, CDCl3)
d 166.7,
164.7, 163.9 (d, 1JCF ¼ 246.7 Hz), 163.7 (d, 1JCF ¼ 246.7 Hz), 160.3 (t,
3JCF ¼ 15.5 Hz), 155.8 (d, JCF ¼ 235.9 Hz), 143.5 (d, JCF ¼ 2.2 Hz),
142.8,142.3,136.1,135.9,129.2,128.9,128.4,127.8,127.8,127.8,126.4,
125.4, 124.4, 116.0 (d, 2C, 2JCF ¼ 22.1 Hz), 113.5 (d, 2C, 3JCF ¼ 15.5 Hz),
1
4
2
2
98.7 (d, 2C, JCF ¼ 28.5 Hz), 97.0 (t, JCF ¼ 25.8 Hz), 80.1, 67.4, 58.0,
57.6, 55.5, 49.8, 49.3, 38.2, 35.5, 32,3, 21.8; HRMS (ESI) m/z 743.2697
([M þ H]þ calcd for C37H42F3N4O7Sþ 743.2721)
5.1.3.38. N-[5-benzylamino-1S-(3,5-difluoro-phenoxymethyl)-2R-hydr-
oxy-4R-methoxy-pentyl]-5-(methanesulfonyl-methyl-amino)-N0-
(1R-phenyl-ethyl)-isophthalamide (2o). Title compound 2o was
obtained from 17 as described for 2n, in 52% yield (21 mg,
20
5.1.3.35. N-[5-benzyl-(2-nitro-benzenesulfonyl)-amino-1S-(3,5-diflu-
oro-phenoxymethyl)-2S-hydroxy-4R-methoxy-pentyl]-5-(methanes-
ulfonyl-methyl-amino)-N0-(1R-phenyl-ethyl)-isophthalamide (16). Title
28
m
mol). 2o vas further purified by LC-MS. [
a
]
D
þ41.5 (c 0.4,
CHCl3); 1H NMR (500 MHz, CD3OD):
d
8.22 (t, 1H, J ¼ 1.6 Hz), 8.04
(m, 2H), 7.42–7.38 (m, 2H), 7.37–7.30 (m, 6H), 7.30–7.22 (m, 2H),
6.63.6.47 (m, 3H), 5.27–5.21 (m, 1H), 4.40–4.35 (m, 1H), 4.32 (d,
2H, J ¼ 5.0 Hz), 4.04–3.99 (m, 1H), 3.90–3.82 (m, 2H), 3.68–3.62
(m, 1H), 3.37 (s, 3H), 3.36 (s, 3H), 2.95 (s, 3H), 2.88 (dd, 1H, J ¼ 4.8,
12.7 Hz), 2.77 (dd, 1H J ¼ 6.0, 12.7 Hz), 2.00–1.93 (m, 1H), 1.63–1.56
(m, 1H), 1.59 (d, 3H, J ¼ 7.1 Hz); 13C NMR (125.8 MHz, CD3OD):
compound 16 was obtained from 14 as described for 2h, in 90%
20
yield (80 mg, 86
(400 MHz, CDCl3)
m
d
mol). [
a
]
D
þ12.2 (c 1.0, CHCl3); 1H NMR
8.08 (m, 1H), 8.00 (m, 1H), 7.96 (m, 1H), 7.63 (m,
3H), 7.40–7.19 (m, 10H), 6.79 (m, 2H), 6.49 (m, 2H), 6.42 (m, 1H),
5.31 (m, 1H) 4.60 (dd, 2H, J ¼ 15.7, 37.1 Hz), 4.22 (m, 1H),4.13–4.00
(m, 3H), 3.51 (m, 1H), 3.37 (m, 5H), 3.18 (s, 3H), 2.87 (s, 3H), 1.67 (m,
1H), 1.61 (d, 3H, J ¼ 7.0 Hz), 1.38 (m, 1H); 13C NMR (100 MHz, CDCl3)
1
1
d
168.8, 167.9, 163.9 (d, JCF ¼ 246.8 Hz), 163.7 (d, JCF ¼ 246.8 Hz),
162.5 (t, 3JCF ¼ 13.8 Hz), 145.1, 143.8, 137.5, 137.2, 129.9, 129.7, 129.6,
1
1
2
d
166.5, 165.0 (d, JCF ¼ 246.4 Hz), 164.8, 162.6 (d, JCF ¼ 246.4 Hz),
129.4, 129.1, 128.8, 128.2, 127.3, 126.0, 99.6 (d, 2C, JCF ¼ 29.1 Hz),
160.3 (t, 3JCF ¼ 13.7 Hz), 147.8142.9, 142.5, 136.3, 135.6, 135.3, 134.0,
97.1 (t, JCF ¼ 26.3 Hz), 78.0, 68.8, 68.7, 57.7, 56.0, 54.1, 52.2, 51.0,
2
133.4, 131.9, 131.4, 128.9, 128.9, 128.4, 128.3, 127.9, 127.8, 127.7,
38.5, 38.3, 35.9; HRMS (ESI) m/z 739.2982 ([M þ H]þ calcd for
2
126.4, 124.8, 123.9, 98.7 (d, 2C, JCF ¼ 28.6 Hz), 96.9 (d,
C38H45F2N4O7Sþ 739.2972)
2JCF ¼ 25.8 Hz), 80.3, 77.4, 68.3, 66.9, 57.7, 52.8, 52.7, 50.8, 48.7, 38.1,
35.9, 35.7, 21.9; HRMS (ESI) m/z 924.2735 ([M þ H]þ calcd for
C44H48F2N5O11Sþ2 924.2754)
5.1.3.39. N-[1S-(3,5-difluoro-phenoxymethyl)-2S-hydroxy-4R-methoxy-
5-(4-methoxy-benzyloxy)-pentyl]-2-[methyl-(2S-methyl-cyclopropyl-(S)-
methyl)-amino]-6-[methyl-(propane-2-sulfonyl)-amino]-isonicotina-
mide (18). Palladium on charcoal (w5%) was added to a solution of
7 (68 mg, 0.16 mmol) in MeOH (3 mL) under argon atmosphere.
The flask was evacuated followed by addition of hydrogen gas
(1 atm). The reaction mixture was stirred over night. The Pd/C was
5.1.3.36. N-[5-benzyl-(2-nitro-benzenesulfonyl)-amino-1S-(3,5-diflu-
oro-phenoxymethyl)-2R-hydroxy-4R-methoxy-pentyl]-5-(methanesul-
fonyl-methyl-amino)-N0-(1R-phenyl-ethyl)-isophthalamide (17). Title
compound 17 was obtained from 15 as described for 2h, in 92%

554
J. Adrian Meredith et al. / European Journal of Medicinal Chemistry 45 (2010) 542–554
filtered off and the filtrate was concentrated to yield the crude
product which was used in the next step without further purifi-
References
[1] K. Blennow, M.J. de Leon, H. Zetterberg, Lancet 368 (2006) 387–403.
[2] H.-W. Klafh, M. Staufenbiel, J. Kornhuber, J. Wiltfang, Brain 129 (2006) 2840–2855.
[3] I.G. Stavrovskaya, B.S. Kristal, Free Radic. Biol. Med. 2005 (2005) 687–697.
[4] C. Milligan, Nat. Med. 6 (2000) 385–386.
[5] A.K. Ghosh, N. Kumaragurubaran, J. Tang, Curr. Top. Med. Chem. 5 (2005)
1609–1622.
[6] T. Iwatsubo, A. Odaka, N. Suzuki, H. Mizusawa, N. Nukina, Y. Ihara, Neuron 13
(1994) 45–53.
[7] C. Lippa, L. Nee, H. Mori, P. St George-Hyslop, Lancet 352 (1998) 1117–1118.
[8] C.J. Pike, D. Burdick, A.J. Walencewicz, C.G. Glabe, C.W. Cotman, J. Neurosci. 13
(1993) 1676–1687.
[9] B.A. Yankner, L.R. Dawes, S. Fisher, L. Villa-Komaroff, M.L. Oster-Granite,
R.L. Neve, Science 245 (1989) 417–420.
[10] J. Hardy, D.J. Selkoe, Science 297 (2002) 353–356.
[11] X. Lin, G. Koelsch, S. Wu, D. Downs, A. Dashti, J. Tang, Proc. Natl. Acad. Sci.
U.S.A. 97 (2000) 1456–1460.
[12] Y. Luo, B. Bolon, S. Kahn, B.D. Bennett, S. Babu-Khan, P. Denis, W. Fan, H. Kha,
J. Zhang, Y. Gong, L. Martin, J.C. Louis, Q. Yan, W.G. Richards, M. Citron,
R. Vassar, Nat. Neurosci. 4 (2001) 231–232.
[13] S.L. Roberds, J. Andersson, G. Basi, M.J. Bienkowski, D.G. Bransetter, K.S. Chen,
S.B. Freedman, N.L. Frigon, D. Games, K. Hu, K. Johnson-Wood,
K.E. Kappenman, T.T. Kawabe, I. Kola, R. Kuehn, M. Lee, W. Liu, R. Motter,
N.F. Nichols, M. Power, D.W. Robertson, D. Schenk, M. Schoor, G.M. Shopp,
M.E. Shuck, S. Sinha, K.A. Svensson, G. Tatsuno, H. Tintrup, J. Wijsman,
S. Wright, L. McConlogue, Hum. Mol. Genet. 10 (2001) 1317–1324.
[14] T. Gou, D.W. Hobbs, Curr. Med. Chem. 13 (2006) 1811–1829.
[15] B. Schmidt, S. Baumann, H.A. Braun, G. Larbig, Curr. Top. Med. Chem. 6 (2006)
377–392.
cation. Carboxylic acid B (23 mg, 65
DCM (2 mL) and PyBOP (34 mg, 65 mol) was added followed by
DIPEA (12 L, 65 mol, 0.4 equiv). After 30 min the crude amine,
dissolved in DCM, was added to the mixture, followed by DIPEA
(12 L, 65 mol). After 2 h, the reaction mixture was diluted with
mmol) was dissolved in dry
m
m
m
m
m
DCM, washed with Na2CO3 (sat.) and NH4Cl (aq). The aqueous
layers were extracted with DCM (3x), and the combined organic
phases were washed with brine, dried (MgSO4), concentrated and
purified by silica column chromatography (toluene–ethyl acetate
gradient 10:1–1:6) to yield the crude compound 18, in 66% yield
(32 mg, 43 mmol). Compound 18 was further purified by LC-MS.
20
[
a]
ꢂ3.4 (c 1.0, MeOH); 1H NMR (400 MHz, CDOD3)
d 7.20 (m,
D
2H), 6.80 (m, 3H), 6.71 (m, 1H), 6.60 (m, 2H), 6.50 (m, 1H), 4.50 (m,
1H), 4.43 (m, 2H), 4.23 (dd, 1H, J ¼ 6.4, 9.8 Hz), 4.16 (dd, 1H, J ¼ 7.1,
9.8 Hz), 4.08 (m, 2H), 3.74 (s, 1H), 3.62 (m, 1H), 3.52 (m, 3H), 3.38
(m, 7H), 3.09 (s, 3H), 1.79 (m, 2H), 1.34 (d, 6H, J ¼ 6.9 Hz), 1.01 (d,
3H, J ¼ 6.0 Hz), 0.77 (m, 1H), 0.69 (m, 1H), 0.43 (m, 1H), 0.22 (m,
1H); 13C NMR (100 MHz, CDOD3)
d
169.7, 163.9 (d, 1JCF ¼ 246.2 Hz),
1
3
163.7 (d, JCF ¼ 246.2 Hz), 162.3 (t, JCF ¼ 13.8 Hz), 160.7, 159.1,
154.8, 146.6, 131.5, 130.5, 114.7, 100.6, 100.1, 99.6 (d, 2C,
2JCF ¼ 28.8 Hz), 97.2 (t, JCF ¼ 26.3 Hz), 79.0, 73.9, 71.7, 68.7, 68.2,
2
57.6, 56.0, 55.6, 54.7, 54.1, 36.8, 36.7, 36.5, 19.1, 18.8, 16.6, 16.6 12.6,
12.0; HRMS (ESI) m/z 749.3419 ([M þ H]þ calcd for C37H51F2N4O8Sþ
749.3390).
[16] Z. Ziora, T. Kimura, Y. Kiso, Drugs Future 31 (2006) 53–63.
[17] J. Eder, U. Hommel, F. Cumin, B. Martoglio, B. Gerhartz, Curr. Pharm. Design 13
(2007) 271–285.
[18] R. Silvestri, Med. Res. Rev. 29 (2009) 295–338.
[19] B. Schmidt, S. Baumann, R. Narlawar, H.A. Braun, G. Larbig, Neurodegenerative
Dis. 3 (2006) 290–297.
[20] J.N. Cumming, U. Iserloh, M.E. Kennedy, Curr. Opin. Drug Discov. Dev. 7 (2004)
536–557.
[21] S.J. Stachel, C.A. Coburn, T.G. Steele, K.G. Jones, E.F. Loutzenhiser, A.R. Gregro,
H.A. Rajapakse, M.-T. Lai, M.-C. Crouthamel, M. Xu, K. Tugusheva,
J.E. Lineberger, B.L. Pietrak, A.S. Espeseth, X.-P. Shi, E. Chen-Dodson,
M.K. Holloway, S. Munshi, A.J. Simon, L. Kuo, J.P. Vacca, J. Med. Chem. 47 (2004)
6447–6450.
[22] M. Ba¨ck, J. Nyhle´n, I. Kvarnstro¨m, S. Appelgren, N. Borkakoti, K. Jansson,
J. Lindberg, S. Nystro¨m, A. Hallberg, Å. Rosenquist, B. Samuelsson, Bioorg. Med.
Chem. 16 (2008) 9471–9486.
[23] C. Bjo¨rklund, S. Oscarson, K. Benkestock, N. Borkakoti, K. Jansson, J. Lindberg,
A. Hallberg, Å. Rosenquist, B. Samuelsson. [unpublished results].
[24] O.T. Schmidt, in: R.L. Whistler, M.L. Woffrom, J.N. BeMiller (Eds.), Methods in
Carbohydrate Chemistry, vol. II, Academic Press, New York, London,1963, p. 318.
[25] K. Oscarsson, M. Lahmann, J. Lindberg, T.U. Kangasmetsa¨, S. Oscarson,
A. Hallberg, B. Samuelsson, Bioorg. Med. Chem. 11 (2003) 1107–1115.
[26] H.R. Jeon, S. Yoon, Y. Shin, J.E.N. Shin, J. Korean Chem. Soc. 41 (1997) 150–156.
[27] P.-O. Johansson, Y. Chen, A.K. Belfrage, M.J. Blackman, I. Kvarnstro¨m,
K. Jansson, L. Vrang, E. Hamelink, A. Hallberg, Å. Rosenquist, B. Samuelsson, J.
Med. Chem. 47 (2004) 3353–3366.
[28] G. Zuccarello, A. Bouzide, I. Kvarnstro¨m, G. Niklasson, S.C.T. Svensson,
M. Brisander, H. Danielsson, U. Nillroth, A. Karle´n, A. Hallberg, B. Classon,
B. Samuelsson, J. Org. Chem. 63 (1998) 4898–4906.
[29] K.M. Sureshan, T. Das, M.S. Shashidhar, R.G. Gonnade, M.M. Bhadbhade, Eur. J.
Org. Chem. (2003) 1035–1041.
5.1.3.40. 2-Ethyl-6-methyl-7,7-dioxo-6,7,8,9-tetrahydro-7l
6-thia-6,9a-
diaza-benzo[cd]azulene-carboxylic acid [1S-(3,5-difluoro-phenoxy-
methyl)-2S-hydroxy-4R-methoxy-5-(4-methoxy-benzyloxy)-pen-
tyl]-amide (19). Palladium on charcoal (w5%) was added to
a solution of 7 (88 mg, 0.20 mmol) in MeOH (3 mL) under argon
atmosphere. The flask was evacuated followed by addition of
hydrogen gas (1 atm). The reaction mixture was stirred over
night. The Pd/C was filtered off and the filtrate was concentrated
to yield the crude product which was used in the next step
without further purification. Carboxylic acid C (26 mg, 84
was dissolved in dry DCM (2 mL) and PyBOP (44 mg, 84
m
m
mol)
mol)
was added followed by DIPEA (15
30 min the crude amine, dissolved in DCM was added to the
mixture, followed by DIPEA (15 L, 84 mol). After 2 h, the
mL, 84 mmol, 0.4 equiv). After
m
m
reaction mixture was diluted with DCM, washed with Na2CO3
(sat.) and NH4Cl (aq). The aqueous layers were extracted with
DCM (3ꢁ), and the combined organic phases were washed with
brine, dried (MgSO4), concentrated and purified by silica column
chromatography (toluene–ethyl acetate gradient 10:1–1:6) to
yield the crude compound 19, in 76% yield (45 mg, 64
m
mol).
20
[30] T. Kinoshita, T. Miwa, J. Clardy, Carbohydr. Res. 143 (1985) 249–255.
[31] R. Johansson, B. Samuelsson, J. Chem. Soc. Perkin Trans. (1984) 2371–2374.
[32] H.H. Jung, P.E. Floreancig, J. Org. Chem. 72 (2007) 7359–7366.
[33] M. Harmata, P. Zheng, C. Huang, M.G. Gomes, W. Ying, K.-O. Ranyanil, G. Balan,
N.L. Calkins, J. Org. Chem. 72 (2007) 683–685.
Compound 19 was further purified by LC-MS. [
a]
D
ꢂ11.7 (c 1.0,
MeOH); 1H NMR (400 MHz, CDOD3)
d
8.08 (d, 1H, J ¼ 1.6 Hz), 7.65
(d, 1H, J ¼ 1.5 Hz), 7.14 (m, 2H), 7.07 (m, 1H), 6.72 (m, 2H), 6.61
(m, 2H), 6.49 (m, 1H), 4.57 (m, 1H), 4.49 (m, 2H) 4.39 (m, 2H),
4.25 (dd, 1H, J ¼ 6.8, 9.9 Hz),4.16 (m, 2H), 3.89 (m, 2H), 3.68 (s,
3H), 3.62 (m, 1H), 3.54 (dd, 1H, J ¼ 3.8, 10.6 Hz), 3.48 (dd, 1H,
J ¼ 4.7, 10.6 Hz), 3.46 (s, 3H), 3.37 (s, 3H), 2.75 (m, 2H), 1.81 (m,
´
[34] E. Frerot, J. Coste, A. Pantaloni, M.-N. Dufour, P. Jouin, Tetrahedron 47 (1991)
259–270.
[35] H. Staudinger, J. Meyer, Helv. Chim. Acta 2 (1919) 635.
[36] S.R. Stauffer, M.G. Stanton, A.R. Gregro, M.A. Steinbeiser, J.R. Shaffer,
P.G. Nantermet, J.C. Barrow, K.E. Rittle, D. Collusi, E.S. Espeseth, M.-T. Lai,
B.L. Pietrak, M.K. Holloway, G.B. McGaughey, S.K. Munshi, J.H. Hochman,
A.J. Simon, H.G. Selnick, S.L. Graham, J.P. Vacca, Bioorg. Med. Chem. Lett. 17
(2007) 1788–1792.
[37] N. Charrier, B. Clarke, L. Cutler, E. Demont, C. Dingwall, R. Dunson, P. East,
J. Hawkins, C. Howes, I. Hussain, P. Jeffrey, G. Maile, R. Matico, J. Mosley,
A. Naylor, A. O’Brien, S. Redshaw, P. Rowland, V. Soleil, K.J. Smith, S. Sweitzer,
P. Theobald, D. Vesey, D.S. Walter, G. Wayne, J. Med. Chem. 51 (2008) 3313–
3317.
2H), 1.31 (t, 3H, J ¼ 7.5 Hz); 13C NMR (100 MHz, CDOD3)
d 170.6,
1
1
165.2 (d, JCF ¼ 244.9 Hz), 165.1 (d, JCF ¼ 244.9 Hz), 162.3 (t,
3JCF ¼ 13.8 Hz), 160.7, 135.6, 131.9, 131.4, 130.4, 129.7, 129.2, 127.7,
2
127.4, 120.9, 119.1, 119.1, 114.6, 99.6 (d, 2C, JCF ¼ 28.7 Hz), 97.1 (t,
2JCF ¼ 26.5 Hz), 79.0, 73.8, 71.7, 68.7 68.3, 57.8, 57.6, 55.6, 53.9,
44.2, 40.2, 36.7, 18.9, 14.9; HRMS (ESI) m/z 724.2486 ([M þ Na]þ
calcd for C35H41F2N3O8SNaþ 724.2475).
[38] PDB ID: 3ELT.
[39] GOLD, version 2.1, Cambridge Crystallographic Data Centre, Cambridge, UK.
[40] Tripos Inc. 1699 South Hanley Rd., St. Louis, Missouri, 63 144, USA.
Acknowledgement
We gratefully thank Medivir AB for financial support.