Synthesis and anti-tuberculosis activity of glycitylamines

Article abstract of DOI:10.1016/j.bmc.2015.12.036

A series of glycitylamines, which were prepared in few steps from readily available carbohydrates, were tested for their ability to inhibit tuberculosis growth in an Alamar Blue BCG colourimetric assay. Several derivatives, including (2R,3R)-1-(hexadecyla

Full text of DOI:10.1016/j.bmc.2015.12.036

Bioorganic & Medicinal Chemistry xxx (2016) xxx–xxx
Contents lists available at ScienceDirect
Bioorganic & Medicinal Chemistry
journal homepage: www.elsevier.com/locate/bmc
Synthesis and anti-tuberculosis activity of glycitylamines
Hilary M. Corkran a,b, Emma M. Dangerfield a,b, Gregory W. Haslett a, Bridget L. Stocker a,b,
,
⇑
Mattie S. M. Timmer a,
⇑
a School of Chemical and Physical Sciences, Victoria University of Wellington, PO Box 600, 6140 Wellington, New Zealand
b Malaghan Institute of Medical Research, PO Box 7060, Wellington, New Zealand
a r t i c l e i n f o
a b s t r a c t
Article history:
A series of glycitylamines, which were prepared in few steps from readily available carbohydrates, were
tested for their ability to inhibit tuberculosis growth in an Alamar Blue BCG colourimetric assay. Several
derivatives, including (2R,3R)-1-(hexadecylamine)pent-4-ene-2,3-diol, (2R,3R)-1-(hexadecylmethy-
Received 26 October 2015
Revised 16 December 2015
Accepted 22 December 2015
Available online xxxx
lamino)pent-4-ene-2,3-diol and 5-deoxy-5-hexadecylmethylamino-
to excellent (44–90%) overall yield and exhibited micromolar (20–41
similar to the first line tuberculosis (TB) drug ethambutol (39 M) in the same assay. The ease and
D
-arabinitol, were prepared in good
l
M) inhibitory activity that was
l
Keywords:
low cost of the synthesis of the glycitylamines offers definite advantages for their use as potential TB
drugs.
Tuberculosis
Carbohydrates
Amines
Ó 2016 Elsevier Ltd. All rights reserved.
Synthesis
Reductive amination
1. Introduction
enzyme responsible for linking the arabinose sugars to the ara-
binogalactan polysaccharide that forms a critical part of the cell
The treatment of infectious diseases that are predominantly
endemic to developing countries requires simple medications that
can be produced in large quantities at low cost. This is particularly
true in the case of medications for the treatment of tuberculosis
(TB), which is a disease that infects more than 9 million new peo-
ple annually and which has a particularly high mortality rate for
those living in developing nations.1 The treatment of TB requires
an extensive period of drug administration and a cocktail of com-
plementary drug treatments due to the persistence of the pathogen
Mycobacterium tuberculosis (M. tuberculosis) inside host cells2 and
increasing multi-drug resistant (MDR) or extreme drug resistant
(XDR) strains of the disease.1 Current first-line drugs for TB include
pyrazinamide (1), ethambutol (2), and isoniazid (3) (Fig. 1),3 and in
addition to their obvious clinical efficacy, another advantage of
these low molecular weight compounds is their ease of synthesis.
Accordingly, we became interested in exploring the potential of
small molecules as therapeutics for the TB so as to add to the arse-
nal of drugs that are required to combat this disease.4
wall of M. tuberculosis,7 there was also some merit in determining
whether simple pryrrolidines (e.g., 5) showed promise as drugs for
TB.8 To this end, we proposed that the glycitylamines could be
readily prepared from their parent furanosides using our previ-
ously published reductive amination methodology,9 with the
pyrrolidines then being synthesised from glycitylamines in two
additional steps via carbamate annulation methodology.10–13 The
ability of the compounds to inhibit the growth of BCG as a model
for M. tuberculosis would then be explored4 so as to ascertain
whether either of these two scaffolds would provide derivatives
that show promise in the treatment of TB.
2. Results and discussion
To commence our studies, we first undertook the synthesis of
the readily accessible glycitylamines
4 and pryrrolidines 5
(Scheme 1) using our previously published protecting-group-free
procedures.10,14,15 Accordingly, pentoses 6 underwent Fischer gly-
cosylation with MeOH before the iodine was installed at the pri-
mary position. A Vasella reductive-amination, involving the use
of Zn, NH4OAc (sat.), NH3 and NaCNBH3 in EtOH, then allowed
for the selective formation of primary amines 4 (R = H),9 which
were subjected to an I2-mediated carbamate annulation10 to give
carbamates 7. Base-mediated hydrolysis then yielded pyrrolidines
5. By starting with different parent sugars, glycitylamines 4a–4d,
pyrrolidines 5a–5d, and the corresponding carbamates 7a–7d were
Inspired by the simplicity of ethambutol (2), we thus sought to
explore whether glycitylamines (e.g., 4, Fig. 2), or derivatives
thereof, might also show promise in the treatment of TB. Moreover,
as ethambutol (2) inhibits arabinosyl transferase (AraT),5,6 the
⇑
Corresponding authors. Tel.: +64 4 463 6529; fax: +64 4 463 5241.
E-mail addresses: bridget.stocker@vuw.ac.nz (B.L. Stocker), mattie.timmer@vuw.
ac.nz (M.S.M. Timmer).
http://dx.doi.org/10.1016/j.bmc.2015.12.036
0968-0896/Ó 2016 Elsevier Ltd. All rights reserved.
Please cite this article in press as: Corkran, H. M.; et al. Bioorg. Med. Chem. (2016), http://dx.doi.org/10.1016/j.bmc.2015.12.036

2
H. M. Corkran et al. / Bioorg. Med. Chem. xxx (2016) xxx–xxx
OH
OH
OH
OH
O
O
NH2
H
N
NH2
NH2
NH2
N
N
NH2
NHNH2
N
H
OH
OH
OH
OH
N
HO
4a
4b
4c
4d
1
2
3
> 3.5 mM
> 3.5 mM
> 3.5 mM
> 3.5 mM
Figure 1. Low molecular weight first-line drugs pyrazinamide (1), ethambutol (2),
and isoniazid (3) for the treatment of M. tuberculosis.
OH
H
OMe
Ph
Ph
H
N
N
Ph
Ph
Ph
H2N
OH
Ph
4e
OMe4f
8
HO
H
N
0.44-0.88 mM
0.80-1.61 mM
1.36-2.73 mM
OH OH
HO
NH2
HO
HO
HO
HO
H
N
H
N
H
N
H
N
OH
HO
OH
4
5
HO
OH
HO
HO
OH
OH
5a
5b
5c
HO 5d
Figure 2. Glycitylamines I (X = H, OH; n = 1, 2) and pyrrolidines II to be tested for
their ability to inhibit the growth of BCG.
> 3.5 mM
> 3.5 mM
> 3.5 mM
> 3.5 mM
O
O
O
O
O
O
O
O
1) MeOH, AcCl
HO
N
N
N
N
OH
2) I2, PPh3, Imid.
O
OH
NHR
3) Zn, NH4OAc
NH3, NaCNBH3
or
Zn, R-NH2,
NaCNBH3
HO
7a
> 3.5 mM
HO
7c
> 3.5 mM
HO
7d
> 3.5 mM
HO 7b
> 3.5 mM
OH
OH
OH
OH
HO
OH
6
4
Figure 3. Glycitylamines 4, pyrrolidines 5 and carbamates 7 tested for their ability
to inhibit the growth of BCG in an Alamar Blue growth inhibition assay. MIC value
determined as the concentration of drug required for 100% inhibition of bacterial
growth.
I2, NaHCO3
(where R = H)
O
HO
H
O
NaOH
N
N
HO
OH
HO
OH
5
7
Table 1
BCG Inhibitory activity of second library of glycitylamines 4
Scheme 1. Synthesis of glycitylamines 4 and pyrrolidines 5.
Entry
1
Product
OH
Yielda (%)
MICb
prepared (Fig. 3), while the use of the secondary amine amin-
odiphenylmethane rather than NH3/NH4OAc gave glycitylamine
4e (R = Ph2CH). The use of methyl 5-deoxy-2,3-di-O-methyl-5-
H
N
Ph
Ph
60
0.44–0.88 mM
OH
4e
OMe
Ph
Ph
iodo-a/b- D-xylofuranoside in the Vasella reductive amination with
H
N
aminodiphenylmethane afforded the fully protected glycitylamine
4f. In this way, a first series of compounds was prepared and sub-
sequently tested for their ability to inhibit growth of BCG in an Ala-
mar Blue growth inhibition assay.4,16,17 As illustrated (Fig. 3),
glycitylamines 4a–4d showed poor inhibitory activity, however
the more hydrophobic derivatives 4e and 4f showed improved,
albeit modest, activity, which was better than the MIC of diphenyl-
methylamine (8) alone. The pyrrolidines 5 and carbamates 7
showed disappointing inhibitory activity with MICs > 3.5 mM,
which may be due to the hydrophilicity of the compounds and
their inability to pass through the highly lipophilic mycobacterial
cell wall.7 While modifications to an arabinose or related pyrro-
lidine scaffold may lead to inhibitors with improved activity, as
has been demonstrated in studies by others,18 the sub-milimolar
inhibitory activity of glycitylamines 4e and the ease of synthesis
of these compounds promoted us to focus on the structure–activity
relationship of these derivatives instead.
To further explore the structure–activity relationship of the
glycitylamines, we next investigated whether the stereochemistry
of the diphenylmethylamine-functionalised derivatives influenced
the inhibitory activity of these compounds. Accordingly, the
growth inhibition activity of the arabinose derivatives 4e and 4f
(entries 1 and 2, respectively, Table 1) was compared to the activity
of the 2S,3R derivative 4g and the 2R,3R derivative 4h (entries 3 and
4), whereby the latter were prepared in 64% and 56% overall yield,
respectively. Unfortunately, 4g and 4h showed no improvement in
inhibitory activity compared to 4e and 4f. The activity of
2
8
0.80–1.61 mM
OMe
4f
OH
H
N
Ph
Ph
Ph
Ph
Ph
3
4
5
6
7
8
9
64
56
57
55
51
25d
60
1.6 mM
1.6 mM
NIc
OH
Ph
Ph
4g
4h
4i
OH
H
N
OH
OH
H
N
OH
OH
H
N
NI
OH
4j
OH
H
N
NI
OH
4k
OH
OH
H
N
2.0 mM
OH
OH
OH
4l
H
N
940 lM
C8H17
OH
4m
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H. M. Corkran et al. / Bioorg. Med. Chem. xxx (2016) xxx–xxx
3
Table 2
Table 1 (continued)
BCG Inhibitory activity of reductive amination products
Entry
Product
OH
Yielda (%)
61
MICb
O
OH
NaCNBH3
R-NH2
OH
OH
H
N
H
N
HO
R
n
10
940
680
l
M
C8H17
C8H17
n
9
OH
4n
OH
H
N
Entry
1
Product
Yield (%)
14
MICa
NIb
11
12
57
31
l
M
OH OH
OH
4o
H
HO
N
OH
H
N
OH
NI
OH
OH
9a
4p
OH OH
H
N
HO
H
N
2
3
43
30
NI
NI
OH
9b
OH OH
13
14
24
14
200
l
M
OH
4q
H
N
OH OH
OH
NI
N
9c
OH OH
OH
OH
H
N
4r
HO
H
N
4
5
6
45
12
10
NI
NI
NI
OH
15
16
17
28
44
47
45 lM
C14H33
C16H33
C16H33
OH
9d
4s
OH OH
OH
H
H
N
N
21–41 lM
OH OH
OH
4t
OH
9e
N
22
39
l
M
M
OH OH
H
N
OH
4u
18
19
20
21
Ethambutol (2)
—
—
—
—
l
OH OH
Cyclododecylamine
Tetradecylamine
Hexadecylamine
680
140–290
130
lM
lM
9f
lM
OH OH
H
N
HO
a
Overall yield starting from
MIC = minimum concentration for 100% growth inhibition in the BCG Alamar
D
-arabinose,
D-ribose or D-xylose.
7
8
26
18
160
680
l
M
OH
b
Blue Assay.
9g
c
NI = No inhibition, MIC > 500
Isolated as a side-product, see Ref. 10.
l
g/mL.
OH OH
d
H
N
lM
OH OH
9h
OH OH
benzylamine derivatives 4i–4k (entries 5–7), which were synthe-
sised from the parent sugars -xylose, -ribose, and -arabinose,
D
D
D
H
N
HO
HO
C16H33
respectively, did not inhibit BCG growth, while the secondary
amine 4l also had reduced inhibitory activity (entry 8). This lack
of activity prompted us to increase the lipophilicity of the com-
pounds and use the more lipophilic octylamine for the reductive
amination (entries 9–11). Octylamine derivatives 4m and 4n
9
78
90
83–170 lM
OH
9i
OH OH
N
C16H33
10
20–40 lM
OH
9j
exhibited MICs of 940
-arabinose, showed the most promising activity (MIC = 680
This prompted us to continue exploring the structure–activity rela-
tionships of derivatives synthesised from -arabinose.
lM, while N-octylamine 4o, prepared from
OH
D
lM).
HO
OH
OH
C16H33
D
11
4c
61 lM
OH OH
Incorporation of the adamantyl group was initially of interest
due to the promising anti-TB activity of ethylenediamine SQ109,
an analogue of EMB, which is now in phase II clinical trials.3,19 Ada-
mantyl-functionalised glycitylamine derivative 4p, however,
exhibited no inhibitory activity (entry 12), while cyclododecyl-
HO
N
OH
9k
a
MIC = minimum concentration for 100% growth inhibition in the BCG Alamar
Blue Assay.
amine derivative 4q had a modest MIC of 200 lM (entry 13).
b
NI = No inhibition, MIC > 500
Side-product initially formed during synthesis of 13i.
l
g/mL.
Formation of the tertiary amine 4r, via use of di-n-butylamine,
gave a glycitylamine with no inhibitory activity (entry 14), while
the preparation of 4s containing the linear C14 alkyl group gave
c
on the nitrogen which was installed via the addition of formalde-
hyde during the reductive amination step (entry 17). Both deriva-
the most promising activity thus far, with an MIC = 45 lM (entry
15). This suggested that linear, rather than cyclic, alkyl groups
led to improved inhibitory activity for this series of compounds.
We thus prepared 4t using hexadecylamine (entry 16), and the
analogous tertiary amine 4u containing an additional methyl group
tives showed good inhibitory activity (4t, MIC = 21–41
MIC = 22 M), which is comparable to that of ethambutol in our
assay (entry 18, MIC = 39 M). Moreover, both 4t and 4u can be
prepared in three-steps and in good (44–47%) overall yield. While
lM; 4u,
l
l
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4
H. M. Corkran et al. / Bioorg. Med. Chem. xxx (2016) xxx–xxx
the inhibitory activity of these compounds is by no means the best
that has been reported for a variety of potential new generation TB
drugs,3 such an easy synthetic route towards active compounds is
advantageous. Finally, to determine that the activity of the alkeny-
lamines was not due to the amines alone, cyclododecylamine
(entry 19), tetradecylamine (entry 20) and hexadecylamine (entry
19) were tested in the growth inhibition assay. All three showed
inhibitory activity that was significantly lower than the corre-
sponding alkenylamine.
Having determined that the more lipophilic alkenylamines were
able to inhibit the growth of BCG at concentrations comparable to
ethambutol, we then set out to determine whether similar glycity-
lamines, which could be prepared in one-step via direct reductive
amination of the parent sugar, would show equivalent or better
hexadexylamine (Aldrich), formaldehyde (Sigma–Aldrich), di-n-
butylamine (Sigma), and tetradecylamine (Aldrich) were used as
received. All chemicals obtained from commercial suppliers were
used without further purification. Zn dust was activated by the care-
ful addition of concentrated H2SO4, followed by decantation and
washing with EtOH (3ꢀ) and hexanes (3ꢀ) and storage under dry
hexanes. All solvents were removed by evaporation under reduced
pressure. Reactions were monitored by TLC analysis on
Macherey–Nagel silica gel coated plastic sheets with detection by
coating with 20% H2SO4 in EtOH followed by charring at ca.
150 °C, by coating with a solution of ninhydrin in EtOH followed
by charring at ca. 150 °C, or by coating with a solution of 5% KMnO4
and 1% NaIO4 in H2O followed by heating. Column chromatography
was performed on Pure Science silica gel (40–63 micron). Dowex
W50-X8 acidic resin (Sigma) was used for ion exchange chromatog-
raphy and HP-20 (Supelco) for reverse phase chromatography.
High-resolution mass spectra were recorded on a Waters Q-TOF
PremierTM Tandem Mass Spectrometer using positive electro-spray
ionisation. Optical rotations were recorded using an Autopol II
(Rudolf Research Analytical) at 589 nm (sodium D-line). Infrared
spectra were recorded as thin films using a Bruker Tensor 27 FTIR
spectrometer, equipped with an Attenuated Total Reflectance
(ATR) sampling accessory, and are reported in wave numbers
(cmꢁ1). Nuclear magnetic resonance spectra were recorded at
20 °C in D2O, CD3OD, pyridine-d5, CDCl3, or DMSO-d6 at (80 °C) using
either a Varian Unity-INOVA operating at 300 MHz or a Varian Unity
operating at 500 MHz. Chemical shifts are given in ppm (d) and were
internally referenced to the residual solvent peak. NMR peak assign-
ments are based on 2D NMR experiments (COSY, HSQC, and HMBC).
Pyrrolidines 5a,10 5b,15 5c14 and 5d,10 carbamates 7a,10 7b,15
7c14 and 7d,10 and glycitylamines 4a,10 4b,15 4c,14 4d,10 4e10 and
4l10 were synthesised according to previously published proce-
inhibitory activity. To this end, D-arabinose was first subjected to
a reductive amination with 1-adamantanamine to give 9a, which
showed no inhibitory activity (entry 1, Table 2). Similarly, the cyclo-
hexyl derivative 9b showed no activity (entry 2), nor did the anal-
ogous compound prepared from L-fucose 9c (entry 3), or the
derivatives containing a cyclooctyl group (9d, 9e, 9f; entries 4–6).
A shift to using cyclododecylamine during the reductive amination,
however, led to the synthesis of derivatives 9g (entry 7) and 9h
(entry 8), whereby the
D
-arabinose derivative 9g showed inhibitory
M) that was slightly better than the analogous
-arabinose derivatives containing the linear
activity (MIC = 160
alkenylamine. The
l
D
C16 alkyl groups showed better activity, that is, secondary amine
9i with an MIC of 83–170 M (entry 9) and the methylated tertiary
amine 9j with an MIC of 20–40 M (entry 10). The activity of the
l
l
latter compound is comparable to ethambutol (2) and the corre-
sponding alkenylamine 4u, and can be synthesised in 90% yield.
Here, better yields of the C16 glycitylamines could be achieved
due the single step synthesis and the more ready separation of
the target product from the unreacted amine starting material by
silica gel flash column chromatography. Finally, the tertiary amine
9k, which was formed as a side-product during the preparation of
9i, was also tested for its inhibitory activity and determined to have
dures. Methyl 5-deoxy-5-iodo-
5-deoxy-5-iodo- /b-
-arabinoside (64% overall yield),14 and
methyl 5-deoxy-5-iodo- /b-
-xylofuranoside (61% overall yield)10
a/b-D
-riboside (67% yield),10 methyl
a
D
a
D
were prepared according to previously published literature proce-
dures and used for the synthesis of the corresponding alkenylami-
nes, as indicated below.
a notable MIC of 61 lM.
3. Conclusion
4.2. Synthesis
We have prepared a number of glycitylamines containing both
lipophilic and hydrophilic groups. Of this family of compounds,
derivatives 4t, 4u and 9j exhibit MIC activities comparable to the
first line drug ethambutol (5) in our assay. While the MIC value of
these new compounds is only in the micro-molar range, they are
easily prepared in few steps and in good to excellent overall yield
and this may be advantageous for the development of cost effective
medications for the treatment of tuberculosis. Mode of action stud-
ies and in vivo analysis to ascertain the full potential of these glyc-
itylamines as TB drugs is currently under investigation.
4.2.1. (2S,3S)-N-Benzhydryl-2,3-dimethoxypent-4-en-1-amine
(4f)
To a solution of methyl 2,3-di-O-methyl-a/b-D
-xylofuranoside20
(301 mg, 1.57 mmol) in THF (20 mL) was added PPh3 (616 mg,
2.35 mmol), iodine (597 mg, 2.35 mmol), and imidazole (214 mg,
3.14 mmol) and the resulting mixture stirred at reflux for 3 h, then
cooled filtered and concentrated in vacuo. The crude reaction
mixture was purified by silica gel flash column chromatography
(Petroleum Ether/EtOAc, 5/1, v/v) to give methyl 5-deoxy-2,3-di-O
-methyl-5-iodo-
0.53 mmol, 34%). Rf = 0.71 (EtOAc); 1H NMR (500 MHz, CDCl3) d
4.98 (d, J1,2 = 4.4 Hz, 1H, H-1 ), 4.87 (s, 1H, H-1b), 4.45 (m, 1H, H-
4b), 4.37 (td, J3,4 = J4,5a = 6.2, J4,5b = 7.3 Hz, 1H, H-4 ), 3.86 (dd,
J2,3 = 5.4, J3,4 = 6.2 Hz, 1H, H-3 ), 3.77 (dd, J1,2 = 4.4, J2,3 = 5.4 Hz,
1H, H-2 ), 3.74–3.73 (m, 2H, H-2b and H-3b), 3.43 (s, 3H, OMe),
3.41 (s, 3H, OMe), 3.36 (dd, J4,5a = 7.3, J5a,5b = 9.7 Hz, 1H, H-5ab),
3.31 (dd, J4,5a = 6.2, J5a,5b = 10.1 Hz, 1H, H-5a ), 3.27 (dd, J4,5b = 7.5,
J5a,5b = 9.7 Hz, 1H, H-5bb), 3.16 (dd, J4,5b = 7.3, J5a,5b = 10.1 Hz, 1H,
H-5b ),
); 13C NMR (125 MHz, CDCl3) d 108.0 (C1b), 100.8 (C1
88.1 (C2b), 85.9 (C2 ), 83.7 (C3b), 83.5 (C3 ), 81.9 (C4b), 77.8
(C4 ), 60.3 (OMe), 55.5 (OMe), 3.6 (C5b), 2.4 (C5 ); HRMS(ESI) m/z
calcd for [C8H15O4I+Na]+: 324.9907, obsd: 324.9915. To a solution
of methyl 5-deoxy-2,3-di-O -methyl-5-iodo- /b- -xylofuranoside
a/b-D-xylofuranoside as a colourless oil (160 mg,
4. Experimental
4.1. General
a
a
a
Unless otherwise stated, all reactions were performed under
atmospheric air. H2O, MeOH (Pure Science), and EtOH (absolute,
Pure Science), were distilled prior to use. EtOAc (Pure Science),
and petroleum ether (Pure Science), HCl (PanReac), AcOH (Ajax
Finechem), DCM (LabServ), 30% aqueous NH3 (J. T. Baker Chemical
Co.), 35% aqueous NH3 (LabServ), AcCl (B&M), NaCNBH3 (Aldrich),
a
a
a
a
a
a
NH4OAc (AnalaR),
D
-arabinose (Sigma–Aldrich),
L-fucose (Sigma–
a
a
Aldrich), -xylose (Sigma–Aldrich),
D
L
-rhamnose (Sigma–Aldrich),
diphenylmethylamine (Aldrich), benzylamine (Aldrich), octylamine
(Aldrich), cyclohexylamine (Riedel), cyclooctylamine (Aldrich),
a
D
(144 mg, 0.48 mmol) in EtOH/H2O (15 mL/0.3 mL) was added
1-adamantylamine
(Sigma),
cyclododecylamine
(Aldrich),
activated Zn (125 mg, 1.91 mmol), aminodiphenylmethane
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H. M. Corkran et al. / Bioorg. Med. Chem. xxx (2016) xxx–xxx
5
(288
AcOH (50
l
L, 307 mg, 1.67 mmol), NaCNBH3 (102 mg, 0.96 mmol), and
L) and the solution was stirred at reflux for 18 h. The
J3,5-trans = 1.3 Hz, 1H, H-5-trans), 5.23 (dt, J4,5-cis = 10.6, J5-cis,5-trans
=
l
J3,5-cis = 1.3 Hz, 1H, H-5-cis), 4.01 (ddt, J3,5-cis = J3,5-trans = 1.2 Hz, J2,3
4.9 Hz, J3,4 = 6.3 Hz, 1H, H-3), 3.93 (ddd, J1b,2 = 3.1 Hz, J2,3 = 4.9 Hz,
J1a,2 = 9.8 Hz, 1H, H-2), 3.15 (dd, J1b,2 = 3.2 Hz, J1a,1b = 13.2 Hz, 1H,
H-1b), 3.10 (dd, J1a,2 = 9.8 Hz, J1a,1b = 13.2 Hz, 1H, H-1a); 13C NMR
(125 MHz, D2O): d 135.2 (C-4), 135.0 (Ci arom.), 134.7 (Ci arom.),
129.4 (CH arom.), 129.4 (CH arom.), 129.3 (CH arom.), 129.3 (CH
arom.), 127.5 (CH arom.), 127.3 (CH arom.), 118.3 (C-5), 73.6
(C-3), 68.9 (C-2), 65.6 (CHPh2), 48.6 (C-1); HRMS(ESI) m/z calcd
for [C18H21O2N+H]+: 284.1651, obsd: 284.1650.
reaction mixture was cooled, and dry loaded onto silica gel for purifi-
cation by flash column chromatography (Petroleum ether/EtOAc,
10/1, v/v) to give alkenylamine 4f as a colourless oil (119 mg,
0.80 mmol, 80%). Rf = 0.53 (EtOAc); [
a]
17 = ꢁ10.0 (c 1.0, CHCl3); 1H
D
NMR (500 MHz, CDCl3) d 7.34–7.10 (m, 10H, aromatics), 5.68 (ddd,
J3,4 = 7.7, J4,5-cis = 10.4, J4,5-trans = 17.3 Hz, 1H, H-4), 5.29
(d, J4,5-trans = 17.3 Hz, 1H, H-5trans), 5.25 (d, J4,5-cis = 10.4 Hz, 1H, H-
5cis), 4.78 (s, 1H, CH-Ph2), 3.77 (dd, J2,3 = 5.9, J3,4 = 7.7 Hz, 1H, H-3),
3.47 (s, 3H, OMe), 3.39 (ddd, J1a,2 = 4.1, J2,3 = 5.9, J1b,2 = 6.8 Hz, 1H,
H-2), 3.30 (s, 3H, OMe), 2.74 (dd, J1a,2 = 4.1, J1a,1b = 12.3 Hz, 1H, H-
1a), 2.60 (dd, J1b,2 = 6.8, J1a,1b = 12.3 Hz, 1H, H-1b); 13C NMR
(125 MHz, CDCl3) d 144.2, 143.8 (aromatics), 134.9 (C4), 128.5,
128.4, 127.6, 127.4, 127.3, 127.0, 126.9, (aromatics), 118.8 (C5),
83.8 (C3), 82.7 (C2), 67.7 (CH-Ph2), 58.9 (OMe), 56.7 (OMe), 47.9
(C1); HRMS(ESI) m/z calcd for [C20H25NO2+H]+: 312.1958, obsd:
312.1959.
4.2.4. (2S,3S)-1-(Benzhydrylamino)pent-4-ene-2,3-diol (4i)
To a solution of methyl 5-deoxy-5-iodo-
0.19 mmol) in EtOH (3.8 mL) was added activated Zn (62 mg,
0.95 mmol), benzylamine (41 L, 0.38 mmol), NaCNBH3 (36 mg,
a/b-D-xyloside (52 mg,
l
0.57 mmol), and AcOH (5 drops). The mixture was stirred at reflux
for 16 h then cooled and the solvent removed under reduced pres-
sure. The resulting oil was co-evaporated three times with MeOH
and HCl (1 M (aq), 2 equiv) to remove the remaining boron. The
product was purified by silica gel flash column chromatography
(DCM/EtOH/MeOH/30% NH3 (aq), 405/2/2/1 ? 305/2/2/1, v/v/v/v)
to give (2S,3S)-1-(benzhydrylamino)pent-4-ene-2,3-diol (4i)
(43 mg, 0.18 mmol, 93%) as a colourless oil. Rf = 0.39 (DCM/EtOH/
4.2.2. (2S,3R)-1-(Benzhydrylamino)pent-4-ene-2,3-diol (4g)
To a solution of methyl 5-deoxy-5-iodo-
0.18 mmol) in EtOH (0.91 mL) was added activated Zn (60 mg,
0.91 mmol), aminodiphenylmethane (78 L, 0.46 mmol) NaCNBH3
a/b-D-riboside (50 mg,
l
(34 mg, 0.55 mmol), and AcOH (5 drops). The mixture was stirred
at reflux for 18 h then cooled and the solvent removed under
reduced pressure. The resulting oil was co-evaporated three times
with MeOH and HCl (1 M (aq), 2 equiv) to remove the remaining
boron and was purified by silica gel flash column chromatography
(DCM/EtOH/MeOH/30% NH3 (aq), 505/2/2/1 ? 305/2/2/1, v/v/v/v)
to give (2S ,3R)-1-((diphenylmethyl)amino)-pent-4-ene-2,3-diol
(4g) as a colourless oil (56 mg, 0.18 mmol, 96%). Rf = 0.62 (DCM/
MeOH/30% NH3 (aq), 35/2/2/1, v/v/v/v); [
a
]
20 = ꢁ55.2° (c 1.0,
D
EtOH); IR (thin film): 3316, 3065, 3035, 3008, 2985, 2957, 2930,
2853, 2795, 1646, 1588, 1500, 1458, 1429, 1323, 1259, 1243,
1212, 1145, 1107, 1093, 1052, 993, 926, 852, 751, 700 cmꢁ1 1H
;
NMR (500 MHz, D2O) 7.49–7.47 (m, 5H, Ph), 5.83 (ddd, J3,4 = 6.3,
J4,5-cis = 10.5, J4,5-trans = 17.1 Hz, 1H, H-4), 5.33 (dd, J4,5-trans
=
17.1, J5-trans,5-cis = 1.0 Hz, 1H, H-5-trans), 5.28 (dd, J4,5-cis = 10.8,
J5-trans,5-cis = 1.0 Hz, 1H, H-5-cis), 4.27 (s, 2H, CH2 N-Bn), 4.07 (dd,
J2,3 = J3,4 = 5.4 Hz, 1H, H-3), 3.89 (ddd, J1b,2 = 2.7, J2,3 = 4.6,
J1a,2 = 9.8 Hz, 1H, H-2), 3.18 (d, J1a,1b = 13.0 Hz, 1H, H-1b), 3.08
(dd, J1a,2 = 10.2, J1a,1b = 13.0 Hz, 1H, H-1a); 13 C NMR (125 MHz,
D2O): d 135.3 (C-4), 130.3 (Ci arom.), 129.8 (CH arom.), 129.6 (CH
arom.), 129.2 (CH arom.), 118.3 (C-5), 73.7 (C-3), 68.9 (C-2), 50.9
(CH2 Bn), 48.5 (C-1); HRMS(ESI) m/z calcd for [C12H17O2N+H]+:
208.1338, obsd: 208.1334.
EtOH/MeOH/30% NH3 (aq), 35/2/2/1, v/v/v/v); [a]
20 = 8.0° (c 1.0,
D
EtOH); IR (thin film): 3345, 3064, 3033, 3009, 2983, 2960, 2926,
2855, 2830, 1646, 1587, 1499, 1455, 1429, 1322, 1288, 1257,
1187, 1144, 1090, 1026, 1001, 990, 926, 891, 840, 793, 760, 746,
696 cmꢁ1 1H NMR (500 MHz, D2O) 7.51–7.42 (m, 10H, Ph), 5.77
;
(ddd, J3,4 = 6.4, J4,5-cis = 10.5, J4,5-trans = 17.1 Hz, 1H, H-4), 5.58 (s,
1H, CHPh2), 5.26 (d, J4,5-trans = 17.8 Hz, 1H, H-5-trans), 5.23
(d, J4,5-cis = 10.7 Hz, 1H, H-5-cis), 4.06 (t, J2,3 = J3,4 = 6.1 Hz, 1H, H-
3), 3.94 (ddd, J1b,2 = 2.5, J2,3 = 4.9, J1a,2 = 9.6 Hz, 1H, H-2), 3.23 (dd,
J1b,2 = 2.2, J1a,1b = 13.2 Hz, 1H, H-1b), 3.04 (dd, J1a,2 = 10.2,
J1a,1b = 13.0 Hz, 1H, H-1a); 13C NMR (125 MHz, D2O): d 135.0 (C-
4), 135.0 (Ci arom.), 134.8 (Ci arom.), 129.4 (CH arom.), 129.4 (CH
arom.), 129.4 (CH arom.), 129.3 (CH arom.), 127.5 (CH arom.),
127.3 (CH arom.), 118.4 (C-5), 74.1 (C-3), 69.0 (C-2), 65.7 (CHPh2),
48.2 (C-1); HRMS(ESI) m/z calcd for [C18H21O2N+H]+: 284.1651,
obsd: 284.1654.
4.2.5. (2S,3R)-1-(Benzylamino)pent-4-ene-2,3-diol (4j)
To a solution of methyl 5-deoxy-5-iodo-
0.19 mmol) in EtOH (0.93 mL) was added activated Zn (60 mg,
0.92 mmol), benzylamine (51 L, 0.46 mmol), NaCNBH3 (35 mg,
a/b-D-riboside (51 mg,
l
0.56 mmol), and AcOH (5 drops). The mixture was stirred at reflux
for 16 h then cooled and the solvent removed under reduced pres-
sure. The resulting oil was co-evaporated three times with MeOH
and HCl (1 M (aq), 2 equiv) to remove the remaining boron and
then purified by silica gel flash column chromatography (DCM/
EtOH/MeOH/30% NH3 (aq), 105/2/2/1 ? 55/2/2/1, v/v/v/v) to give
(2S,3R)-1-(benzylamino)pent-4-ene-2,3-diol (4j) as a colourless
oil (37 mg, 0.15 mmol, 82%). Rf = 0.30 (DCM/EtOH/MeOH/30% NH3
4.2.3. (2R,3R)-1-(Benzhydrylamino)pent-4-ene-2,3-diol (4h)
To
(52 mg, 0.19 mmol) in EtOH (0.99 mL) was added activated Zn
(65 mg, 0.99 mmol), aminodiphenylmethane (85 L, 0.50 mmol),
NaCNBH3 (37 mg, 0.60 mmol), and AcOH (5 drops). The mixture
was stirred at reflux for 18 h then cooled and the solvent removed
under reduced pressure. The resulting oil was co-evaporated three
times with MeOH and HCl (1 M (aq), 2 equiv) to remove the
remaining boron and purified by silica gel flash column chro-
a solution of methyl 5-deoxy-5-iodo-a/b-D-arabinoside
l
(aq), 35/2/2/1, v/v/v/v); [
3331, 3065, 3034, 3009, 2984, 2957, 2928, 2787, 2760, 1645,
a
]
20 = ꢁ5.7° (c 1.0, EtOH); IR (thin film):
D
1587, 1500, 1457, 1429, 1322, 1261, 1211, 1184, 1144, 1089,
1072, 1028, 992, 927, 848, 751, 698 cmꢁ1 1H NMR (500 MHz,
;
D2O) 7.50–7.47 (m, 5H, Ph), 5.84 (ddd, J3,4 = 6.4, J4,5-cis = 10.5,
J4,5-trans = 17.1 Hz, 1H, H-4), 5.32 (d, J4,5-trans = 18.8 Hz, 1H, H-5-
trans), 5.29 (d, J4,5-cis = 11.0 Hz, 1H, H-5-cis), 4.29 (d, Ja,b = 13.2 Hz,
1H, CH-a N-Bn), 4.26 (d, Ja,b = 13.2 Hz, 1H, CH-b N-Bn), 4.11 (t,
J2,3 = J3,4 = 6.1 Hz, 1H, H-3), 3.89 (ddd, J1b,2 = 2.9, J2,3 = 5.4,
J1a,2 = 9.8 Hz, 1H, H-2), 3.26 (dd, J1a,1b = 12.9, J1b,2 = 2.6 Hz, 1H, H-
1b), 3.03 (dd, J1a,2 = 10.0, J1a,1b = 13.0 Hz, 1H, H-1a); 13 C NMR
(125 MHz, D2O): d 135.1 (C-4), 130.3 (Ci arom.), 129.8 (CH arom.),
129.6 (CH arom.), 129.2 (CH arom.), 118.4 (C-5), 74.1 (C-3), 68.9
(C-2), 51.0 (CH2 Bn), 48.1 (C-1); HRMS(ESI) m /z calcd for
[C12H17O2N+H]+: 208.1338, obsd: 208.1335.
matography (DCM/EtOH/MeOH/30% NH3
305/2/2/1, v/v/v/v) to give diol 4h as a colourless oil (55 mg,
0.17 mmol, 87%). Rf = 0.66 (DCM/EtOH/MeOH/30% NH3 (aq),
(aq), 505/2/2/1 ?
35/2/2/1, v/v/v/v); [
a]
20 = ꢁ1.8° (c 0.5, EtOH); IR (thin film):
D
3335, 3064, 3033, 3009, 2923, 2857, 2826, 1645, 1602, 1586,
1499, 1455, 1409, 1186, 1106, 1053, 1033, 1004, 991, 924, 878,
843, 792, 760, 746, 705 cmꢁ1 1H NMR (500 MHz, D2O) 7.52–7.41
;
(m, 10H, Ph), 5.77 (ddd, J3,4 = 6.3, J4,5-cis = 10.6, J4,5-trans = 17.2 Hz,
1H, H-4), 5.58 (s, 1H, CHPh2), 5.28 (dt, J4,5-trans = 17.5, J5-cis,5-trans
=
Please cite this article in press as: Corkran, H. M.; et al. Bioorg. Med. Chem. (2016), http://dx.doi.org/10.1016/j.bmc.2015.12.036

6
H. M. Corkran et al. / Bioorg. Med. Chem. xxx (2016) xxx–xxx
4.2.6. (2R,3R)-1-(Benzylamino)pent-4-ene-2,3-diol (4k)
To solution of methyl 5-deoxy-5-iodo- /b- -arabinoside
(49 mg, 0.18 mmol) in EtOH (0.88 mL) was added activated Zn
(58 mg, 0.88 mmol), benzylamine (48 L, 0.44 mmol), NaCNBH3
(33 mg, 0.53 mmol), and AcOH (5 drops). The mixture was stirred
at reflux for 16 h then cooled and the solvent removed under
reduced pressure. The resulting oil was co-evaporated three times
with MeOH and HCl (1 M (aq), 2 equiv) to remove the remaining
boron and purified by silica gel flash column chromatography
(DCM/EtOH/MeOH/30% NH3 (aq), 305/2/2/1 ? 105/2/2/1, v/v/v/v)
and HCl (1 M (aq), 2 equiv) to remove the remaining boron and
the product purified by silica gel flash column chromatography
(DCM/EtOH/MeOH/30% NH3 (aq), 305/2/2/1 ? 105/2/2/1, v/v/v/v)
to give diol 4n as a colourless oil (48 mg, 0.18 mmol, 91%).
Rf = 0.18 (DCM/EtOH/MeOH/30% NH3 (aq), 35/2/2/1, v/v/v/v);
a
a
D
l
[a]
20 = ꢁ3.3° (c 1.0, EtOH); IR (thin film): 3342, 3120, 3049, 3024,
D
2957, 2924, 2855, 2806, 1646, 1590, 1445, 1401, 1314, 1230,
1148, 1098, 1027, 994, 924, 844, 760, 723, 703 cmꢁ1 1H NMR
;
(500 MHz, D2O) 5.90 (ddd, J3,4 = 6.4, J4,5-cis = 10.5, J4,5-trans = 17.1 Hz,
1H, H-4), 5.38 (d, J4,5-trans = 17.1 Hz, 1H, H-5-trans), 5.34
(d, J4,5-cis = 10.5 Hz, 1H, H-5-cis), 4.15 (dd, J2,3 = 5.6, J3,4 = 6.3 Hz,
1H, H-3), 3.91 (ddd, J1b,2 = 2.7, J2,3 = 5.4, J1a,2 = 9.8 Hz, 1H, H-2),
3.27 (dd, J1b,2 = 2.7, J1a,1b = 13.2 Hz, 1H, H-1b), 3.07 (t, J = 7.3 Hz,
2H, CH2N octyl), 3.03 (dd, J1a,2 = 10.0, J1a,1b = 13.0 Hz, 1H, H-1a),
1.69 (q, J = 7.1 Hz, 2H, CH2 N-octyl), 1.38–1.27 (m, 10H, 5ꢀCH2 N-
octyl), 0.86 (t, J = 7.1 Hz, 3H, CH3 N-octyl); 13 C NMR (125 MHz,
D2O): d 135.3 (C-4), 118.5 (C-5), 74.1 (C-3), 69.1 (C-2), 48.8 (C-1),
47.8 (CH2 octyl), 30.9 (CH2 octyl), 28.1 (CH2 octyl), 28.1 (CH2 octyl),
25.6 (CH2 octyl), 25.2 (CH2 octyl), 21.9 (CH2 octyl), 13.4 (CH3 octyl);
HRMS(ESI) m/z calcd for [C13H27O2N+H]+: 230.2120, obsd:
230.2118.
to give (2R,3R)-1-(benzylamino)pent-4-ene-2,3-diol (4k) as
colourless oil (34 mg, 0.14 mmol, 79%). Rf = 0.28 (DCM/EtOH/
MeOH/30% NH3 (aq), 35/2/2/1, v/v/v/v); [
20 = 35.7° (c 1.0, EtOH);
a
a
]
D
IR (thin film): 3330, 3091, 3065, 3035, 3008, 2985, 2958, 2928,
2792, 2759, 1647, 1588, 1500, 1458, 1428, 1323, 1259, 1212,
1184, 1144, 1099, 1052, 1026, 992, 926, 854, 750, 699 cmꢁ1 1H
;
NMR (500 MHz, D2O) 7.49–7.46 (m, 5H, Ph), 5.83 (ddd, J3,4 = 6.5,
J4,5-cis = 10.7, J4,5-trans = 17.3 Hz, 1H, H-4), 5.34 (dt, J4,5-trans = 17.3,
J5-cis,5-trans = J3,5-trans = 1.2 Hz, 1H, H-5-trans), 5.28 (dt, J4,5-cis
=
10.5, J5-cis,5-trans = J3,5-cis = 1.2 Hz, 1H, H-5-cis), 4.27 (s, 2H, CH2 N-
Bn), 4.07 (ddt, J2,3 = 4.9 Hz, J3,4 = 6.4 Hz, J3,5-cis = J3,5-trans = 1.2 Hz,
1H, H-3), 3.89 (ddd, J1b,2 = 3.0, J2,3 = 4.9, J1a,2 = 10.1 Hz, 1H, H-2),
3.19 (dd, J1b,2 = 2.9, J1a,1b = 12.9 Hz, 1H, H-1b), 3.08 (dd,
J1a,2 = 10.0, J1a,1b = 12.9 Hz, 1H, H-1a); 13C NMR (125 MHz, D2O): d
135.3 (C-4), 130.3 (Ci arom.), 129.8 (CH arom.), 129.7 (CH arom.),
129.2 (CH arom.), 118.3 (C-5), 73.7 (C-3), 68.9 (C-2), 51.0 (CH2
Bn), 48.6 (C-1); HRMS(ESI) m/z calcd for [C12H17O2N+H]+:
208.1338, obsd: 208.1338.
4.2.9. (2R,3R)-1-(Octylamino)pent-4-ene-2,3-diol (4o)
To
(51 mg, 0.19 mmol) in EtOH (3.6 mL) was added activated Zn
(61 mg, 0.93 mmol), octylamine (61 L, 0.37 mmol), NaCNBH3
a solution of methyl 5-deoxy-5-iodo-a/b-D-arabinoside
l
(35 mg, 0.56 mmol), and AcOH (5 drops). The mixture was stirred
at reflux for 18 h then cooled and the solvent removed under
reduced pressure. The resulting oil was co-evaporated three times
with MeOH and HCl (1 M (aq), 2 equiv) to remove the remaining
boron and the product purified by silica gel flash column chro-
4.2.7. (2S,3S)-1-(Octylamino)pent-4-ene-2,3-diol (4m)
To a solution of methyl 5-deoxy-5-iodo-
0.20430 mmol) in EtOH (4.1 mL) was added activated Zn (67 mg,
1.02 mmol), octylamine (68 L, 0.41 mmol), NaCNBH3 (39 mg,
a/b-D-xyloside (56 mg,
matography (DCM/EtOH/MeOH/30% NH3
(aq), 405/2/2/1 ?
l
305/2/2/1, v/v/v/v) to give (2R,3R)-1-(octylamino)pent-4-ene-2,3-
diol (4o) as a colourless oil (44 mg, 0.17 mmol, 89%). Rf = 0.19
0.61 mmol), and AcOH (5 drops). The mixture was stirred at reflux
for 17 h then cooled and the solvent removed under reduced pres-
sure. The resulting oil was co-evaporated three times with MeOH
and HCl (1 M (aq), 2 equiv) to remove the remaining boron and
purified by silica gel flash column chromatography (DCM/EtOH/
MeOH/30% NH3 (aq), 305/2/2/1 ? 105/2/2/1, v/v/v/v) to give (2S,
3S)-1-(octylamino)-pent-4-ene-2,3-diol (4m) as a colourless oil
(54 mg, 0.20316 mmol, 99%). Rf = 0.21 (DCM/EtOH/MeOH/30%
(DCM/EtOH/MeOH/30% NH3 (aq), 35/2/2/1, v/v/v/v); [a]
20 = 41.6°
D
(c 1.0, EtOH); IR (thin film): 3313, 3091, 3077, 3014, 2958, 2924,
2855, 2797, 1646, 1593, 1455, 1406, 1315, 1250, 1152, 1102,
1090, 1055, 991, 922, 852, 767, 723, 699 cmꢁ1
;
1H NMR
(500 MHz, D2O) 5.86 (ddd, J3,4 = 6.4, J4,5-cis = 10.5, J4,5-trans = 17.1 Hz,
1H, H-4), 5.36 (dt, J4,5-trans = 17.3, J5-cis,5-trans = J3,5-trans = 1.2 Hz, 1H,
H-5-trans), 5.30 (dt, J4,5-cis = 10.7, J5-cis,5-trans = J3,5-cis = 1.2 Hz, 1H,
H-5-cis), 4.10 (dd, J2,3 = 5.0, J3,4 = 6.4 Hz, 1H, H-3), 3.88 (ddd,
J1b,2 = 3.0, J2,3 = 4.9, J1a,2 = 10.3 Hz, 1H, H-2), 3.17 (dd, J1b,2 = 2.9,
J1a,1b = 13.0 Hz, 1H, H-1b), 3.06 (dd, J1a,2 = 10.2, J1a,1b = 12.9 Hz,
1H, H-1a), 3.04 (t, JCH2,CH2 = 7.3 Hz, 2H, CH2N octyl), 1.66 (q,
J = 7.4 Hz, 2H, CH2 N-octyl), 1.36–1.24 (m, 10H, 5ꢀCH2 N-octyl),
0.83 (t, J = 7.1 Hz, 3H, CH3 N-octyl); 13C NMR (125 MHz, D2O): d
135.3 (C-4), 118.3 (C-5), 73.7 (C-3), 69.0 (C-2), 49.1 (C-1), 47.7
(CH2 octyl), 30.9 (CH2 octyl), 28.0 (CH2 octyl), 28.0 (CH2 octyl),
25.5 (CH2 octyl), 25.1 (CH2 octyl), 21.9 (CH2 octyl), 13.3 (CH3 octyl);
HRMS(ESI) m/z calcd for [C13H27O2N+H]+: 230.2120, obsd:
230.2117.
NH3 (aq), 35/2/2/1, v/v/v/v); [
a
]
20 = ꢁ42.0° (c 1.0, EtOH); IR (thin
D
film): 3321, 3089, 3015, 2957, 2924, 2855, 2800, 1647, 1593,
1455, 1407, 1378, 1315, 1285, 1250, 1152, 1140, 1103, 1091,
1051, 992, 923, 853, 755, 723, 699 cmꢁ1 1H NMR (500 MHz,
;
D2O) 5.86 (ddd, J3,4 = 6.6, J4,5-cis = 10.8, J4,5-trans = 17.4 Hz, 1H, H-4),
5.36 (dd, J4,5-trans = 17.3, J5-trans,5-cis = 1.5 Hz, 1H, H-5-trans), 5.30
(dd, J4,5-cis = 10.5, J5-trans,5-cis = 1.2 Hz, 1H, H-5-cis), 4.10 (t,
J2,3 = J3,4 = 6.4 Hz, 1H, H-3), 3.88 (ddd, J1b,2 = 3.0, J2,3 = 4.9,
J1a,2 = 10.1 Hz, 1H, H-2), 3.17 (dd, J1b,2 = 2.5, J1a,1b = 12.9 Hz, 1H,
H-1b), 3.09–3.03 (m, 3H, H-1a and CH2N octyl), 1.66 (q,
J = 7.3 Hz, 2H, CH2 N-octyl), 1.36–1.25 (m, 10H, 5ꢀCH2 N-octyl),
0.83 (t, J = 7.1 Hz, 3H, CH3 N-octyl); 13C NMR (125 MHz, D2O): d
135.3 (C-4), 118.3 (C-5), 73.7 (C-3), 69.0 (C-2), 49.1 (C-1), 47.7
(CH2 octyl), 30.9 (CH2 octyl), 28.0 (CH2 octyl), 28.0 (CH2 octyl),
25.5 (CH2 octyl), 25.1 (CH2 octyl), 21.9 (CH2 octyl), 13.3 (CH3 octyl);
HRMS(ESI) m /z calcd for [C13H27O2N+H]+: 230.2120, obsd:
230.2122.
4.2.10. (2R,3R)-1-(Adamantan-1-ylamino)pent-4-ene-2,3-diol
(4p)
To
a solution of methyl 5-deoxy-5-iodo-a/b-D-arabinoside
(134 mg, 0.49 mmol) in EtOH (9.8 mL) was added 1-adaman-
tanamine (96 mg, 0.64 mmol), activated Zn (159 mg, 2.43 mmol),
NaCNBH3 (93 mg, 1.48 mmol), water (0.3 mL) and AcOH
(0.15 mL). The mixture was stirred under reflux overnight (18 h),
then cooled, filtered and the solvent removed under reduced pres-
sure. The product was dry loaded on silica and purified by gradient
silica gel flash column chromatography (DCM/EtOH/MeOH/35%
NH3 (aq), 305/2/2/1 to 5/2/2/1, v/v/v/v). Further purification by
C8 reverse phase chromatography (H2O to H2O/MeOH, 1/4, v/v) fol-
lowed by the addition of 1 M HCl(aq) and concentration gave
4.2.8. (2S,3R)-1-(Octylamino)pent-4-ene-2,3-diol (4n)
To a solution of methyl 5-deoxy-5-iodo-
0.20 mmol) in EtOH (0.99 mL) was added activated Zn (65 mg,
0.99 mmol), octylamine (82 L, 0.49 mmol), NaCNBH3 (38 mg,
a/b-D-riboside (54 mg,
l
0.59 mmol), and AcOH (5 drops). The mixture was stirred at reflux
for 16 h then cooled and the solvent removed under reduced pres-
sure. The resulting oil was co-evaporated three times with MeOH
Please cite this article in press as: Corkran, H. M.; et al. Bioorg. Med. Chem. (2016), http://dx.doi.org/10.1016/j.bmc.2015.12.036

H. M. Corkran et al. / Bioorg. Med. Chem. xxx (2016) xxx–xxx
7
(2R,3R)-1-(adamantan-1-ylamino)pent-4-ene-2,3-diol hydrochlo-
ride (4p) as a white solid (70 mg, 0.24 mmol, 49%). Rf = 0.42
6H, H-40); 13C NMR (125 MHz, CDCl3) d 137.5 (C4), 116.5 (C5),
74.4 (C3), 69.2 (C2), 57.0 (C1), 54.2 (C10), 28.8 (C20), 20.5 (C30),
14.0 (C40); HRMS(ESI) m/z calcd for [C13H27O2N+H]+: 230.2115,
obsd: 230.2122.
(DCM/EtOH/MeOH/30% NH3 (aq), 15/2/2/1, v/v/v/v); [
(c 1.0, EtOH); IR (film) 3364, 2915, 2855, 1644, 1454, 1365, 1310,
1071, 1022, 938 cmꢁ1 1H NMR (500 MHz, D2O) d 5.90 (ddd,
a]
25 = +24.9
D
.
J3,4 = 5.9, J4,5-cis = 10.6, J4,5-trans = 17.2 Hz, 1H, H-4), 5.39
(d, J4,5-trans = 17.2 Hz, 1H, H-5-trans), 5.33 (d, J4,5-cis = 10.6 Hz, 1H,
H-5-cis), 4.14 (dd, J2,3 = 5.1, J3,4 = 5.9 Hz, 1H, H-3), 3.84 (ddd,
J1a,2 = 2.6, J2,3 = 5.1, J1b,2 = 10.2 Hz, 1H, H-2), 3.18 (dd, J1a,2 = 2.6,
J1a,1b = 12.7 Hz, 1H, H-1a), 3.04 (dd, J1b,2 = 10.2, J1a,1b = 12.7 Hz,
1H, H-1b), 2.22–2.17 (m, 3H, H-30), 1.95–1.85 (m, 6H, H-20), 1.75
4.2.13. (2R,3R)-1-(Tetradecylamino)pent-4-ene-2,3-diol (4s)
To
a solution of methyl 5-deoxy-5-iodo-a/b-D-arabinoside
(146 mg, 0.53 mmol) in EtOH (10 mL) was added tetradecylamine
(136 mg, 0.64 mmol), activated Zn (291 mg, 4.45 mmol), NaCNBH3
(101 mg, 1.60 mmol), water (0.3 mL), and AcOH (0.15 mL). The
mixture was stirred under reflux overnight (18 h), then cooled,
filtered and the solvent removed under reduced pressure. The pro-
duct was dry loaded on silica and purified by gradient silica gel
(d, J 4 a,4 b = 12.5 Hz, 3H, H-40a), 1.66 (d, J 4 a,4 b = 12.5 Hz, 3H, H-
40b); 13C NMR (125 MHz, D2O) d 135.4 (C4), 118.4 (C5), 73.9 (C3),
69.6 (C2), 57.9 (C10), 41.7 (C1), 37.8 (C20), 34.8 (C40), 28.8 (C30);
HRMS(ESI) m /z calcd for [C15H25O2N+H]+: 252.1958, obsd:
252.1961.
0
0
0
0
flash chromatography (DCM/EtOH/MeOH/35% NH3
(aq),
305/2/2/1 to 5/2/2/1, v/v/v/v) to yield diol 4s as a white solid
(74 mg, 0.24 mmol, 44%). Rf = 0.65 (DCM/EtOH/MeOH/30% NH3
(aq), 35/2/2/1, v/v/v/v); [
3275, 2954, 2916, 2847, 1646, 1468, 1420, 1326, 1252, 1162,
1119, 1095, 1053, 1030, 987, 926, 859 cmꢁ1 1H NMR (500 MHz,
a]
21 = +0.3 (c 0.66, EtOH); IR (film) 3380,
D
4.2.11. (2R,3R)-1-(Cyclododecylamino)pent-4-ene-2,3-diol (4q)
To
a
solution of methyl 5-deoxy-5-iodo-
a/b-
D-arabinoside
.
(89.1 mg, 0.33 mmol) in EtOH (6.6 mL) was added cyclododecy-
lamine (65.5 mg, 0.36 mmol), activated Zn (195 mg, 2.98 mmol),
NaCNBH3 (62 mg, 0.98 mmol), water (0.2 mL), and AcOH
(0.1 mL). The mixture was stirred under reflux overnight (18 h),
then cooled, filtered, and the solvent removed under reduced pres-
sure. The product was dry loaded on silica and purified by gradient
silica gel flash column chromatography (DCM/EtOH/MeOH/35%
NH3 (aq), 305/2/2/1 to 5/2/2/1, v/v/v/v) to yield (2R,3R)-1-(cyclodo-
decylamino)pent-4-ene-2,3-diol hydrochloride (4q) as a white
solid (38 mg, 0.12 mmol, 37%). Rf = 0.69 (DCM/EtOH/MeOH/30%
CD3OD) d 5.93 (ddd, J3,4 = 6.3, J4,5-cis = 10.6, J4,5-trans = 17.2 Hz, 1H,
H-4), 5.32 (ddd, J3,5-trans = 1.5, J5-cis,5-trans = 1.6, J4,5-trans = 17.2 Hz,
1H, H-5-trans), 5.19 (ddd, J3,5-cis = 1.5, J5-cis,5-trans = 1.6, J4,5-cis = 10.6
Hz, 1H, H-5-cis), 3.99 (ddd, J3,5 = 1.5, J2,3 = 5.2, J3,4 = 6.3 Hz, 1H, H-
3), 3.65 (ddd, J1a,2 = 3.7 Hz, J2,3 = 5.2, J1b,2 = 8.8 Hz, 1H, H-2), 2.72
10a,20
(dd, J1a,2 = 3.7, J1a,1b = 12.2 Hz, 1H, H-1a), 2.63 (dt, J
= 7.6, J
1 a,1 b = 11.7 Hz, 1H, H-10a), 2.61 (dd, J1b,2 = 8.8, J1a,1b = 12.2 Hz, 1H,
0
0
H-1b), 2.58 (dt, J1 b,2 = 7.6, J1 a,1 b = 11.7 Hz, 1H, H-10b), 1.56–1.49
(m, 2H,
H-20), 1.36–1.27 (m, 22H, H-30–H-130), 0.91 (t, J
= 7.0 Hz, 3H, H-140); 13C NMR (125 MHz, CD3OD) d 139.0
0
0
0
0
130,140
NH3 (aq), 35/2/2/1, v/v/v/v); [
a]
21 = ꢁ0.7 (c 1.0, EtOH); IR (film)
(C4), 116.7 (C5), 76.1 (C3), 73.5 (C2), 52.6 (C1), 50.6 (C10), 33.0,
30.7, 30.7, 30.6, 30.6. 30.6, 30.4, 29.5, 28.3, 28.2, 27.9, 23.7
(C20-C130), 14.3 (C140); HRMS(ESI) m/z calcd for [C19H39O2N+H]+:
314.3054, obsd: 314.3058.
D
3335, 2932, 2863, 1642, 1471, 1446, 1096, 1025, 926, 719 cmꢁ1
.
1H NMR (500 MHz, CD3OD) d 5.98 (ddd, J3,4 = 5.6, J4,5-cis = 10.7,
J4,5-trans = 17.1 Hz, 1H, H-4), 5.40 (d, J4,5-trans = 17.1 Hz, 1H,
H-5-trans), 5.26 (d, J4,5-cis = 10.5 Hz, 1H, H-5-cis), 4.16–4.11 (m,
1H, H-3), 3.93–3.86 (m, 1H, H-2), 3.34–3.28 (m, 1H, H-10), 3.18
(d, J1a,1b = 12.7 Hz, 1H, H-1a), 3.05 (dd, J1a,2 = 9.7, J1a,1b = 12.7 Hz,
1H, H-1b), 1.80 (ddd, J = 7.3 Hz, J = 14.3 Hz, J = 22.0 Hz, 2H, H-20a),
1.75–1.64 (m, 2H, H-20b), 1.58–1.34 (m, 18H, H-30–H-70); 13C
NMR (125 MHz, CD3OD) d 138.1 (C4), 117.3 (C5), 75.1 (C3), 70.5
(C2), 57.9 (C10), 49.3 (C1), 27.3, 26.7, 25.7, 25.6, 25.4, 24.0, 24.0,
4.2.14. (2R,3R)-1-(Hexadecylamino)pent-4-ene,2,3-diol (4t)
To
a solution of methyl 5-deoxy-5-iodo-a/b-D-arabinoside
(102 mg, 0.37 mmol) in EtOH (7.4 mL), was added hexadecylamine
(110 mg, 0.44 mmol), activated Zn (97 mg, 1.49 mmol), NaCNBH3
(69 mg, 1.10 mmol), water (0.22 mL), and AcOH (0.11 mL). The
mixture was stirred under reflux overnight (18 h), then cooled, fil-
tered and the solvent removed under reduced pressure. The pro-
duct was dry loaded on silica and purified by gradient silica gel
23.9, 23.9, 21.6, 21.3
(C20-C60) HRMS(ESI) m/z calcd for
[C17H33O2N+H]+: 284.2584, obsd: 284.2584.
flash chromatography (DCM/EtOH/MeOH/35% NH3
(aq),
4.2.12. (2R,3R)-1-(Dibutylamino)pent-4-ene-2,3-diol (4r)
305/2/2/1 to 5/2/2/1, v/v/v/v) to yeild (2R,3R)-1-(hexadecy-
lamino)pent-4-ene,2,3-diol hydrochloride (4t) as as a white solid
(96 mg, 0.25 mmol, 69%). Rf = 0.19 (DCM/EtOH/MeOH/30% NH3
To
(100.2 mg, 0.37 mmol) in EtOH (7.4 mL) was added di-n-buty-
lamine (74 L, 0.44 mmol), activated Zn (25 mg, 3.75 mmol),
a solution of methyl 5-deoxy-5-iodo-a/b-D-arabinoside
l
(aq), 35/2/2/1, v/v/v/v); [
2955, 2919, 2850, 1739, 1650, 1470, 1373, 1146, 1016 cmꢁ1
NMR (500 MHz, CD3OD) 5.96 (ddd, J3,4 = 5.2, J4,5-cis = 10.7,
a]
22 = +28.4 (c 1.0, EtOH); IR (film) 3352,
D
NaCNBH3 (71 mg, 1.14 mmol), water (0.22 mL), and AcOH
(0.11 mL). The mixture was stirred under reflux overnight (18 h),
then cooled, filtered, and the solvent removed under reduced pres-
sure. The product was dry loaded on silica and purified by gradient
silica gel flash chromatography (DCM/EtOH/MeOH/35% NH3 (aq),
305/2/2/1 to 5/2/2/1, v/v/v/v) to yield 4r as a white solid (19 mg,
0.08 mmol, 22%). Rf = 0.90 (DCM/EtOH/MeOH/30% NH3 (aq),
.
1H
d
J4,5-trans = 16.9 Hz, 1H, H-4), 5.38 (d, J4,5-trans = 16.9 Hz, 1H, H-5-
trans), 5.25 (dd, J5-cis,5-trans = 0.9, J4,5-cis = 10.7 Hz, 1H, H-5-cis), 4.09
(dd, J2,3 = 4.4, J3,4 = 5.2 Hz, 1H, H-3), 3.87 (dd, J1,2 = 1.7,
J2,3 = 4.4 Hz, 1H, H-2), 3.12 (dd, J1a,2 = 2.5 Hz, J1a,1b = 12.6 Hz, 1H,
H-1a), 3.06–2.95 (m, 3H, H-1b, H-10), 1.75–1.66 (m, 2H, H-20),
a]
20 = +48.3 (c 0.82, CHCl3); IR (film) 3370,
D
1.42–1.24 (m, 26H, H-30–H-150), 0.90 (t, J 15 ,16 = 6.7 Hz, 3H, H-
160); 13C NMR (125 MHz, CD3OD) d 138.1 (C4), 117.3 (C5), 75.0
(C3), 70.5 (C2), 50.8 (C1), 48.5 (C10), 33.1, 30.8, 30.8, 30.8, 30.7,
30.6, 30.5, 30.2, 27.6, 27.0, 23.7 (C20-C150), 14.5 (C160); HRMS
(ESI) m/z calcd for [C21H43O2N+H]+: 342.3367, obsd: 342.3371.
0
0
35/2/2/1, v/v/v/v); [
3086, 2957, 2931, 2872, 2863, 2821, 1644, 1467, 1378, 1305,
1267, 1163, 1069, 994, 921, 735 cmꢁ1 1H NMR (500 MHz, CDCl3)
.
d 5.92 (ddd, J3,4 = 5.8, J4,5-cis = 10.5, J4,5-trans = 17.3 Hz, 1H, H-4),
5.38 (ddd, J3,5-trans = 1.5, J5-cis,5-trans = 1.5, J4,5-trans = 17.3 Hz, 1H, H-
5-trans), 5.24 (ddd, J3,5-cis = 1.4, J5-cis,5-trans = 1.5, J4,5-cis = 10.5 Hz,
1H, H-5-cis), 3.99 (ddd, J3,5 = 1.4, J2,3 = 4.2, J3,4 = 5.8 Hz, 1H, H-3),
3.65 (ddd, J2,3 = 4.2, J1b,2 = 4.3, J1a,2 = 8.6 Hz, 1H, H-2), 2.62 (ddd,
4.2.15. (2R,3R)-1-(Hexadecyl(methyl)amino)pent-4-ene (4u)
To
0.04 mmol) in MeOH (1 mL) and AcOH (0.05 mL), was added 37%
aqueous formaldehyde (60 L, 0.8 mmol), and NaCNBH3
(75.6 mg, 1.2 mmol). The solution was stirred under reflux for
24 h with additional formaldehyde (100 L) and NaCNBH3
a
solution of hexadecylalkenylamine 4t (15.8 mg,
J = 8.7, J = 11.7, J 1 a,1 b = 15.9 Hz, 2H, H-10a), 2.59 (dd, J1a,2 = 8.6,
J1a,1b = 12.9 Hz, 1H, H-1a), 2.54 (dd, J1b,2 = 4.3, J1a,1b = 12.9 Hz, 1H,
0
0
l
H-1b), 2.47 (ddd, J = 5.2, J = 8.6, J1 a,1 b = 15.9 Hz, 2H, H-10b), 1.53–
0
0
1.40 (m, 4H, H-20), 1.38–1.25 (m, 4H, H-30), 0.92 (t, J3 ,4 = 7.3 Hz,
l
0
0
Please cite this article in press as: Corkran, H. M.; et al. Bioorg. Med. Chem. (2016), http://dx.doi.org/10.1016/j.bmc.2015.12.036

8
H. M. Corkran et al. / Bioorg. Med. Chem. xxx (2016) xxx–xxx
(100 mg) added after 15 h and 21 h, respectively. For workup, an
additional 1 mL of formaldehyde was added to remove the excess
reducing agent, and the mixture was concentrated in vacuo. Purifi-
cation by C8 reverse phase column chromatography, eluting in
MeOH gave alkenylamine 4u as a white solid (11 mg, 0.03 mmol,
73%). Rf = 0.69 (DCM/EtOH/MeOH/30% NH3 (aq), 35/2/2/1, v/v/v/
1.18–1.11 (m, 2H, H-40b); 13C NMR (125 MHz, D2O) d 71.0 (C3),
70.4 (C2), 65.9 (C4), 62.7 (C1), 57.5 (C10), 47.3 (C5), 28.9, 28.5
(C20), 24.4 (C40), 23.9, 23.9 (C30); HRMS(ESI) m/z calcd for
[C11H23O4N+H]+: 234.1700, obsd: 234.1705.
4.2.18. 1-Cyclohexylamino-1,6-dideoxy-L-galactitol (9c)
v); [
1658, 1643, 1466, 1379, 1266, 1072, 994, 921 cmꢁ1
(500 MHz, CD3OD) 5.97 (ddd, J3,4 = 5.0, J4,5-cis = 10.5,
a
]
D
22 = +28.0 (c 0.5, MeOH); IR (film) 3373, 3156, 2923, 2853,
To a solution of -fucose (100 mg, 0.63 mmol) in EtOH (3 mL),
L
.
1H NMR
was added cyclohexylamine (54 mg, 0.55 mmol), NaCNBH3
(113 mg, 1.8 mmol), water (0.3 mL), and AcOH (0.04 mL). The mix-
ture was stirred at room temperature for 3 d. The solvent was then
removed under reduced pressure, and the residue purified by
Dowex-H+ cation exchange chromatography, eluting with 5–35%
NH3 in H2O. Subsequent silica gradient flash chromatography
(DCM/EtOH/MeOH/35% NH3 (aq), 105/2/2/1 to 15/2/2/1, v/v/v/v)
d
J4,5-trans = 17.5 Hz, 1H, H-4), 5.33 (d, J4,5-trans = 17.5 Hz, 1H, H-5-
trans), 5.20 (d, J4,5-cis = 10.5 Hz, 1H, H-5-cis), 4.04 (dd, J2,3 = 4.4,
J3,4 = 5.0 Hz, 1H, H-3), 3.72 (ddd, J2,3 = 4.4, J = 7.5, J = 9.5 Hz, 1H,H-
2),2.53(dd, J = 4.6, J = 13.0 Hz, 1H, H-1a), 2.45–2.40 (m, 3H, H-1b,
H-10), 2.28 (s, 3H, N-Me), 1.52–1.48 (m, 2H, H-20), 1.37–1.22 (m,
26H, H-30–H-150), 0.90 (t, J 15 ,16 = 7.0 Hz, 3H, H-160); 13C NMR
(125 MHz, CD3OD) d 139.1 (C4), 116.4 (C5), 76.1 (C3), 71.7 (C2),
61.1 (C1), 59.5 (C10), 43.0 (N-Me), 33.1, 30.8, 30.8, 30.8, 30.7,
30.7, 30.5, 28.5, 28.0, 23.7 (C20-C150), 14.4 (C160); HRMS(ESI) m/z
calcd for [C22H45O2N+H]+: 356.3523, obsd: 356.3523.
afforded 1-cyclohexylamino-1,6-dideoxy-
0.17 mmol, 30%) as a white solid. Rf = 0.40 (DCM/EtOH/MeOH/30%
NH3 (aq), 15/2/2/1, v/v/v/v); [
25 = +10.6 (c 0.86, EtOH); IR (film)
3351, 2948, 2836, 1651, 1450, 1410, 1114, 1017 cmꢁ1
D
-galactitol (9c) (41 mg,
0
0
a
]
D
.
1H NMR
(500 MHz, D2O) d 4.14 (ddd, J2,3 = 1.6, J1b,2 = 4.5, J1a,2 = 8.5 Hz, 1H,
H-2), 4.10 (qd, J4,5 = 1.7, J5,6 = 6.7 Hz, 1H, H-5), 3.58 (dd, J2,3 = 1.6,
J3,4 = 9.3 Hz, 1H, H-3), 3.47 (dd, J4,5 = 1.7, J3,4 = 9.3 Hz, 1H, H-4),
3.11 (dd, J1a,2 = 8.5, J1a,1b = 13.0 Hz, 1H, H-1a), 3.09 (dd, J1b,2 = 4.5,
J1a,1b = 13.0 Hz, 1H, H-1b), 3.00–2.94 (m, 1H, H-10), 2.07–2.01 (m,
2H, H-20a), 1.84–1.77 (m, 2H, H-30a), 1.69–1.64 (m, 1H, H-40a),
1.34–1.28 (m, 4H, H-20b, H-30b), 1.25 (d, J5,6 = 6.7 Hz, 3H, H-6),
1.23–1.12 (m, 1H, H-40b); 13C NMR (125 MHz, D2O) d 72.7 (C4),
70.9 (C3), 67.1 (C2), 65.8 (C5), 57.0 (C10), 47.7 (C1), 29.9, 29.6
(C20), 24.8 (C40), 24.1, 24.1 (C30), 18.6 (C6); HRMS(ESI) m/z calcd
for [C12H25O4N+H]+: 248.1856, obsd: 248.1866.
4.2.16. 5-(Adamant-1-ylamino)-5-deoxy-D-arabinitol (9a)
To a solution of -arabinose (100 mg, 0.67 mmol) in EtOH
D
(3.4 mL), was added 1-adamantanamine (121 mg, 0.80 mmol),
NaCNBH3 (126 mg, 2 mmol), water (0.34 mL), and AcOH
(0.05 mL). The mixture was stirred at room temperature for 20 h.
The solvent was then removed under reduced pressure, and the
residue purified by Dowex-H+ cation exchange chromatography,
eluting with 5–35% NH3 in EtOH/H2O. Further purification by C8
reverse phase chromatography (H2O to H2O/MeOH, 1/1, v/v) and
then gradient flash chromatography (DCM/EtOH/MeOH/35% NH3
(aq), 75/2/2/1 to 35/2/2/1, v/v/v/v), followed by treatment with
1 M HCl(aq) and then concentration afforded 5-(adamant-1-yla-
4.2.19. 5-Cyclooctylamino-5-deoxy-D-arabinitol (9d)
To a solution of -arabinose (100 mg, 0.67 mmol) in EtOH
D
mino)-5-deoxy-
(30 mg, 0.09 mmol, 14%). Rf = 0.09 (DCM/EtOH/MeOH/30% NH3
(aq), 15/2/2/1, v/v/v/v); [
25 = +6.6 (c 0.27, EtOH); IR (film) 3365,
2916, 2854, 1645, 1455, 1365, 1310, 1071, 1020 cmꢁ1
(500 MHz, D2O) d 4.13 (td, J3,4 = 1.7, J4,5 = 6.4 Hz, 1H, H-4), 3.82
(dd, J1a,2 = 2.8, J1a,1b = 11.7 Hz, 1H, H-1a), 3.71 (ddd, J1a,2 = 2.8,
J1b,2 = 5.9, J2,3 = 8.9 Hz, 1H, H-2), 3.65 (dd, J1b,2 = 5.9, J1a,1b = 11.7 -
Hz, 1H, H-1b), 3.50 (dd, J3,4 = 1.7, J2,3 = 8.9 Hz, 1H, H-3), 3.18 (d,
J4,5 = 6.4 Hz, 1H, H-5), 2.19 (br s, 3H, H-30), 1.95–1.87 (m, 6H, H-
D
-arabinitol hydrochloride (9a) as a white solid
(13.3 mL), was added cyclooctylamine (77 mg, 0.60 mmol),
NaCNBH3 (126 mg, 2 mmol), and water (0.4 mL). The mixture
was stirred at room temperature for 4 h. The solvent was then
removed under reduced pressure, and the residue purified by
Dowex-H+ cation exchange chromatography eluting with 5–35%
a]
D
.
1H NMR
NH3 in EtOH/H2O to give 5-cyclooctylamino-5-deoxy-
(9d) as a white solid (70 mg, 0.27 mmol, 45%). Rf = 0.18 (DCM/
EtOH/MeOH/30% NH3 (aq), 15/2/2/1, v/v/v/v); [
25 = +4.9 (c 1.0,
EtOH); IR (film) 3348, 2925, 2857, 1642, 1469, 1446, 1054,
879 cmꢁ1 1H NMR (500 MHz, CD3OD)
3.97 (td, J3,4 = 2.1,
D-arabinitol
a]
D
20), 1.74 (d, J 4 a,4 b = 12.2 Hz, 3H, H-40a), 1.65 (d, J 4 a,4 b = 12.2 Hz,
3H, H-40b); 13C NMR (125 MHz, D2O) d 71.1 (C3), 70.4 (C2), 66.2
(C4), 62.7 (C1), 57.8 (C10), 42.7 (C5), 37.8 (C20), 34.8 (C40), 28.7
(C30); HRMS(ESI) m/z calcd for [C15H27O4N+H]+: 286.2013, obsd:
286.2022.
.
d
0
0
0
0
J4,5 = 6.2 Hz, 1H, H-4), 3.79 (dd, J1a,2 = 3.2, J1a,1b = 10.9 Hz, 1H, H-
1a), 3.66 (ddd, J1a,2 = 3.2, J1b,2 = 5.8, J2,3 = 8.0 Hz, 1H, H-2), 3.62
(dd, J1b,2 = 5.8, J1a,1b = 10.9 Hz, 1H, H-1b), 3.42 (dd, J3,4 = 2.1,
J2,3 = 8.0 Hz, 1H, H-3), 2.83 (d, J4,5 = 6.2 Hz, 2H, H-5), 2.83–2.79
(m, 1H, H-10), 1.87–1.73 (m, 4H, H-20-50), 1.61–1.59 (m, 4H, H-20-
50), 1.59–1.46 (m, 6H, H-20-50); 13C NMR (125 MHz, CD3OD) d
74.3 (C3), 73.0 (C2), 69.7 (C4), 64.9 (C1), 59.2 (C10), 51.0 (C5),
4.2.17. 5-Cyclohexylamino-5-deoxy-D-arabinitol (9b)
To a solution of -arabinose (156 mg, 1.0 mmol) in EtOH
D
(20 mL), was added cyclohexylamine (89 mg, 0.9 mmol), NaCNBH3
(194 mg, 3 mmol), water (0.5 mL), and AcOH (0.07 mL). The mix-
ture was stirred at room temperature for 23 h. The solvent was
then removed under reduced pressure and the residue purified
by Dowex-H+ cation exchange chromatography, eluting with 25%
aqueous NH3, to give a white solid (90 mg, 0.39 mmol, 43%) which
was then treated with 1 M HCl(aq) and concentrated to afford 5-
33.0, 28.1, 26.9, 25.3
(C20-50); HRMS(ESI) m/z calcd for
[C13H27O4N+H]+: 262.2013, obsd: 262.2016.
4.2.20. 1-Cyclooctylamino-1,6-dideoxy-L-galactitol (9e)
To a solution of -fucose (100 mg, 0.63 mmol) in EtOH (3 mL),
L
was added cyclooctylamine (84 mg, 0.66 mmol), NaCNBH3
(113 mg, 1.8 mmol), water (0.3 mL), and AcOH (0.04 mL). The mix-
ture was stirred at room temperature for 22 h. The solvent was
then removed under reduced pressure, and the residue purified
by Dowex-H+ cation exchange chromatography, eluting with 5–
35% NH3 in H2O. Gradient flash chromatography (DCM/EtOH/
MeOH/35% NH3 (aq), 105/2/2/1 to 55/2/2/1, v/v/v/v), followed by
treatment with 1 M HCl and concentration afforded 1-cycloocty-
cyclohexylamino-5-deoxy-
white solid. Rf = 0.16 (DCM/EtOH/MeOH/30% NH3 (aq), 15/2/2/1,
v/v/v/v); [
25 = +12.8 (c 1.0, EtOH); IR (film) 3324, 2935, 2858,
1636, 1455, 1032, 896 cmꢁ1
D-arabinitol hydrochloride (9b) as a
a
]
D
.
1H NMR (500 MHz, D2O) d 4.17 (dd,
J4,5 = 3.2, J3,4 = 8.8 Hz, 1H, H-4), 3.80 (dd, J1a,2 = 2.6 Hz, J1a,1b = 11.7
Hz, 1H, H-1a), 3.70 (ddd, J1a,2 = 2.6, J1b,2 = 5.8, J2,3 = 8.7 Hz, 1H, H-
2), 3.63 (dd, J1b,2 = 5.8, J1a,1b = 11.7 Hz, 1H, H-1b), 3.48 (d,
J3,4 = 8.8 Hz, 1H, H-3), 3.25–3.19 (m, 2H, H-5), 3.18–3.11 (m, 1H,
H-10), 2.10–2.04 (m, 2H, H-20a), 1.82 (d, J = 12.5 Hz, 2H, H-30a),
1.64 (d, J = 13.2 Hz, 2H, H-40a), 1.42–1.25 (m, 4H, H-20b, H-30b)
lamino-1,6-dideoxy-
solid (19 mg, 0.07 mmol, 12%). Rf = 0.16 (DCM/EtOH/MeOH/30%
NH3 (aq), 15/2/2/1, v/v/v/v); [
25 = +10.5 (c 1.0, EtOH); IR (film)
3355, 2928, 2858, 1642, 1450, 1285, 1052, 1023 cmꢁ1
D-galactitol hydrochloride (9e) as a white
a
]
D
.
1H NMR
Please cite this article in press as: Corkran, H. M.; et al. Bioorg. Med. Chem. (2016), http://dx.doi.org/10.1016/j.bmc.2015.12.036

H. M. Corkran et al. / Bioorg. Med. Chem. xxx (2016) xxx–xxx
9
(500 MHz, D2O) d 4.13 (ddd, J2,3 = 1.6, J1b,2 = 4.4, J1a,2 = 9.1 Hz, 1H,
4.2.23. 1-Cyclododecylamino-1,6-dideoxy-L-galactitol (9h)
H-2), 4.01 (qd, J4,5 = 1.7, J5,6 = 6.6 Hz, 1H, H-5), 3.51 (dd, J2,3 = 1.6,
To a solution of -fucose (165 mg, 1.0 mmol) in EtOH (20 mL),
L
J3,4 = 9.4 Hz, 1H, H-3), 3.39 (dd, J4,5 = 1.7, J3,4 = 9.4 Hz, 1H, H-4),
was added cyclododecylamine (294 mg, 1.5 mmol), NaCNBH3
(189 mg, 3.0 mmol), water (0.5 mL), and AcOH (0.07 mL). The mix-
ture was stirred at room temperature for 7 d. The solvent was then
removed under reduced pressure, and the residue purified by
Dowex-H+ cation exchange chromatography eluting with 5–35%
NH3 in EtOH/H2O. Reverse phase chromatography (HP20 beads,
10–100% MeOH), afforded a white solid (59.8 mg, 0.18 mmol,
18%) that was subsequently treated with 1 M HCl(aq) and concen-
3.35 (tt, J 1 ,2 a = 3.5, J 1 ,2 b = 9.3 Hz, 1H, H-10), 3.18 (dd, J1a,2 = 9.1,
J1a,1b = 13.2 Hz, 1H, H-1a), 3.14 (dd, J1b,2 = 4.4, J1a,1b = 13.2 Hz,
1H, H-1b), 1.92–1.85 (m, 2H, H-20a), 1.73–1.65 (m, 4H, H-20b, H-
30a), 1.60–1.40 (m, 7H, H-30b-50a), 1.37–1.30 (m, 1H, H-50b), 1.17
(d, J5,6 = 6.6 Hz, 3H, H-6); 13C NMR (125 MHz, D2O) d 72.6 (C4),
70.7 (C3), 66.2 (C2), 65.6 (C5), 59.0 (C10), 47.8 (C1), 29.2, 28.7
(C20), 25.6, 25.6 (C40), 25.2 (C50), 23.3 (C30), 18.6 (C6); HRMS(ESI)
m/z calcd for [C14H29O4N+H]+: 276.2169, obsd: 276.2170.
0
0
0
0
trated to provide 1-cyclododecylamino-1,6-dideoxy-
hydrochloride (9h). Rf = 0.29 (DCM/EtOH/MeOH/30% NH3 (aq),
5/2/2/1, v/v/v/v); [
25 = +6.0 (c 0.29, EtOH); IR (film) 3366, 2928,
2859, 1637, 1471, 1448, 1414, 1052, 996 cmꢁ1
(500 MHz, D2O) 4.21 (t, J1,2 = 6.3 Hz, 1H, H-2), 4.07
(q, J5,6 = 6.6 Hz, 1H, H-5), 3.56 (d, J3,4 = 9.5 Hz, 1H, H-3), 3.44 (d,
J3,4 = 9.5 Hz, 1H, H-4), 3.32–3.38 (m, 1H,
H-10), 3.24 (d,
D-galactitol
a]
D
4.2.21. 1-Cyclooctylamino-1,6-dideoxy-L-mannitol (9f)
.
1H NMR
To a solution of -rhamnose (100 mg, 0.63 mmol) in EtOH
L
d
(5.5 mL), was added cyclooctylamine (62 mg, 0.49 mmol),
NaCNBH3 (104 mg, 1.7 mmol), water (0.3 mL), and AcOH
(0.15 mL). The mixture was stirred at room temperature for 19 h.
The solvent was then removed under reduced pressure, and the
residue purified by Dowex-H+ cation exchange chromatography,
eluting with 5–35% NH3 in EtOH/H2O. Silica gradient flash chro-
matography (DCM/EtOH/MeOH/35% NH3 (aq), 105/2/2/1 to
5/2/2/1, v/v/v/v), followed by treatment with 1 M HCl(aq) and con-
J1,2 = 6.3 Hz, 2H, H-1), 1.74–1.84 (m, 2H, H-20a), 1.63–1.73 (m, 2H,
H-20b), 1.32–1.51 (m, 18H, H-30-H-70), 1.22 (d, J5,6 = 6.6 Hz, 3H, H-
6); 13C NMR (125 MHz, D2O) d 72.5 (C4), 70.7 (C3), 66.0 (C2),
65.6 (C5), 56.9 (C10), 48.0 (C1), 25.7, 25.3, 24.0, 24.0, 23.9, 22.5,
22.3, 22.3, 22.2, 19.9, 19.7(C20-C70), 18.5 (C6); HRMS(ESI) m/z calcd
for [C18H37O4N+H]+: 332.2795, obsd: 332.2802.
centration afforded 1-cyclooctylamino-1,6-dideoxy-L-mannitol
hydrochloride (9f) as a white solid (16 mg, 0.05 mmol, 10%).
Rf = 0.36 (DCM/EtOH/MeOH/30% NH3 (aq), 5/2/2/1, v/v/v/v);
4.2.24. 5-Deoxy-5-hexadecylamino-
D-arabinitol (9i)
To a solution of -arabinose (100 mg, 0.67 mmol) in EtOH
D
[a]
25 = +1.3 (c 0.38, EtOH); IR (film) 2930, 2851, 1641, 1450,
D
(46.6 mL), was added hexadecylamine (1.66 g, 6.9 mmol),
NaCNBH3 (62.1 mg, 1.0 mmol), and AcOH (2.6 mL). The mixture
was stirred at 50 °C for 4 h then the solvent was removed under
reduced pressure. The residue was co-evaporated with 2ꢀ 25 mL
of 5% HCl in methanol (w/v), then purified by gradient silica
gel flash chromatography (DCM/EtOH/MeOH/35% NH3 (aq),
105/2/2/1 to 15/2/2/1, v/v/v/v) to give HCl salt 9i as a white solid
(195 mg, 0.52 mmol, 78%). Rf = 0.40 (DCM/EtOH/MeOH/35% NH3
(aq), 5/2/2/1, v/v/v/v); IR (film) 3268, 2917, 2849, 1467,1093,
1306, 1071, 1019 cmꢁ1 1H NMR (500 MHz, D2O) d 3.94 (ddd,
.
J1a,2 = 3.0, J2,3 = 8.2, J1b,2 = 9.5 Hz, 1H, H-2), 3.82 (dq, J5,6 = 6.3,
J4,5 = 8.1 Hz, 1H, H-5), 3.78 (dd, J3,4 = 1.5, J2,3 = 8.2 Hz, 1H, H-3),
3.51 (dd, J3,4 = 1.5, J4,5 = 8.1 Hz, 1H, H-4), 3.46–3.40 (m, 1H, H-10),
3.41 (dd, J1a,2 = 3.0, J1a,1b = 13.0 Hz, 1H, H-1a), 3.11 (dd, J1b,2 = 9.5,
J1a,1b = 13.0 Hz, 1H, H-1b), 1.98–1.90 (m, 2H, H-20a), 1.81–1.70
(m, 4H, H-20b, H-30a), 1.67–1.45 (m, 7H, H-30b-50a), 1.42–1.35 (m,
1H, H-50b), 1.25 (d, J5,6 = 6.3 Hz, 3H, H-6); 13C NMR (125 MHz,
D2O) d 72.9 (C4), 70.9 (C3), 66.9 (C2), 66.8 (C5), 59.0 (C10), 47.5
(C1), 29.3, 28.6 (C20), 25.6, 25.6 (C40), 25.1 (C50), 23.3, 23.3 (C30),
18.9 (C6); HRMS(ESI) m/z calcd for [C14H29O4N+H]+: 276.2169,
obsd: 276.2175.
1040, 878 cmꢁ1 1H NMR (300 MHz, DMSO-d6, 80 °C) d 3.78–3.71
.
(m, 1H, H-4), 3.60 (dd, J1a,2 = 3.7, J1a,1b = 10.4 Hz, 1H, H-1a), 3.50
(ddd, J1a,2 = 3.7, J1b,2 = 5.7, J2,3 = 7.5 Hz, 1H, H-2), 3.42 (dd,
J1b,2 = 5.7, J1a,1b = 10.4 Hz, 1H, H-1b), 3.31 (dd, J3,4 = 2.5,
J2,3 = 7.5 Hz, 1H, H-3), 2.71–2.62 (m, 2H, H-5), 2.59–2.52 (m, 2H,
H-10), 1.47–1.36 (m, 2H, H-20), 1.36–1.21 (m, 26H, H-30-H-150),
0.87 (t, J150,160 = 6.8 Hz, 3H, H-160); 13C NMR (75 MHz, DMSO-d6,
80 °C) d 72.9 (C3), 71.7 (C2), 68.5 (C4), 63.5 (C1), 52.4 (C5), 48.9
(C5), 30.9, 28.6 (br. s), 26.4, 21.6 (C20-C150), 13.5 (C160); HRMS
(ESI) m/z calcd for [C21H45O4N+H]+: 376.3421, obsd: 376.3421.
4.2.22. 5-Cyclododecylamino-5-deoxy-D-arabinitol (9g)
To a solution of -arabinose (100 mg, 0.67 mmol) in EtOH
D
(13.3 mL), was added cyclododecylamine (243 mg, 1.33 mmol),
NaCNBH3 (126 mg, 2 mmol), and water (0.4 mL). The mixture
was stirred at room temperature for 5 h then the solvent removed
under reduced pressure. The product was purified first by gradient
flash chromatography (DCM/EtOH/MeOH/35% NH3 (aq), 305/2/2/1
to 5/2/2/1, v/v/v/v), followed Dowex-H+ cation exchange (5–35%
NH3 in EtOH/H2O) to give a white solid, 9g, which was treated with
1 M HCl(aq) and concentrated to give the HCl salt (61 mg,
0.17 mmol, 26%). Rf = 0.24 (DCM/EtOH/MeOH/30% NH3 (aq),
4.2.25. 5-Deoxy-5-hexadecylmethylamino-
D-arabinitol (9j)
To a solution of -arabinose (101.5 mg, 0.67 mmol) in MeOH
D
(46.6 mL), was added hexadecylamine (1.65 g, 6.8 mmol),
NaCNBH3 (64.8 mg, 1 mmol), and AcOH (2.6 mL). The mixture
was stirred at 45 °C for 4 h before 1 mL of 37% aqueous formalde-
hyde (13.3 mmol) was added. After a further 3 h, additional
NaCNBH3 (840 mg, 13.3 mmol) was added and the reaction was
left to stir overnight (16 h), then the solvent was removed under
reduced pressure. The residue was co-evaporated with 2ꢀ 25 mL
of 5% HCl in methanol (w/v), then the residue was purified by gra-
dient silica gel flash chromatography (DCM/EtOH/MeOH/35% NH3
(aq), 105/2/2/1 to 85/2/2/1, v/v/v/v) affording 9j as a white solid
that was converted into the HCl salt by the addition of HCl
(1.2 M, aq) and concentration (236 mg, 0.61 mmol, 90%). Rf = 0.57
15/2/2/1, v/v/v/v); [
2929, 2862, 1576, 1471, 1446, 1074 cmꢁ1
CD3OD) d 4.20–4.17 (m, 1H, H-4), 3.80 (dd, J1a,2 = 2.8, J1a,1b
a]
25 = +8.1 (c 0.93, EtOH); IR (film) 3330,
D
.
1H NMR (500 MHz,
=
10.5 Hz, 1H, H-1a), 3.68 (ddd, J1a,2 = 2.8, J1b,2 = 5.4, J2,3 = 8.1 Hz,
1H, H-2), 3.65 (dd, J1b,2 = 5.4, J1a,1b = 10.5 Hz, 1H, H-1b), 3.47 (dd,
0
0
0
0
J3,4 = 1.7, J2,3 = 8.1 Hz, 1H, H-3), 3.31 (dt, J1 ,2 a = 1.6, J1 ,2 b = 3.2 Hz,
1H, H-10), 3.21 (dd, J4,5a = 8.4, J5a,5b = 12.8 Hz, 1H, H-5a), 3.18 (dd,
J4,5b = 4.4, J5a,5b = 12.8 Hz, 1H, H-5b), 1.91–1.78 (m, 2H, H-20a),
1.74–1.66 (m, 2H, H-20b), 1.57–1.35 (m, 18H, H-30-H-70); 13C
NMR (125 MHz, CD3OD) d 73.3 (C3), 72.7 (C2), 67.4 (C4), 64.6
(C1), 57.9 (C10), 49.7 (C5), 27.3, 26.7 (C20), 25.7, 25.6, 25.4, 24.1,
(DCM/EtOH/MeOH/35% NH3 (aq), 5/2/2/1, v/v/v/v); [
1.0, MeOH); IR (film) 3333, 2953, 2918, 2850, 1467, 1081, 1033,
736 cmꢁ1 1H NMR (300 MHz, CD3OD) d 4.29 (d, J = 9.3 Hz, 1H, H-
4), 3.80 (app. d, J = 8.0 Hz, 1H, H-1a), 3.72–3.60 (m, 2H, H-1b and
a]
28 = +8.0 (c
D
24.0, 23.9, 21.6, 21.3
(C30-C70); HRMS(ESI) m/z calcd for
.
[C17H35O4N+H]+: 318.2639, obsd: 318.2648.
Please cite this article in press as: Corkran, H. M.; et al. Bioorg. Med. Chem. (2016), http://dx.doi.org/10.1016/j.bmc.2015.12.036

10
H. M. Corkran et al. / Bioorg. Med. Chem. xxx (2016) xxx–xxx
H-2), 3.48–3.34 (m, 2H, H-3 and H-5a), 3.26–3.10 (m, 3H, H-10 and
H-5b), 2.93 (s, 3H, N-Me), 1.84–1.69 (m, 2H, H-20), 1.46–1.24 (m,
had any inherent redox potential which might affect readouts,
200 L solutions of compounds and PBS were added to individual
control wells. Control wells of bacteria only, and PBS medium only
were also prepared. 100 L of BCG solution was added to all wells
except the PBS only and compound only wells, and the plate was
incubated at 37 °C and 5% CO2 for 7 days. Alamar Blue dye
l
26H, H-30-H-150), 0.90 (t, J 15 ,16 = 6.6 Hz, 3H, H-160); 13C NMR
(125 MHz, CD3OD) d 71.4 (C3), 71.2 (C2), 64.7 (C4), 63.3 (C1),
55.5 (C5), 56.6 (C10), 40.2 (N-Me), 31.7, 29.4, 29.4, 29.3, 29.2,
29.1, 28.9, 26.3, 22.3 (C30-C150), 23.8 (C20), 13.1 (C160); HRMS
(ESI) m/z calcd for [C22H47O4N+H]+: 390.3578, obsd: 390.3594.
0
0
l
(20 lL) was then added to all test wells on day 7, and the plate
was incubated for another 24 h before fluorescence was measured
(excitation at 544 nm, emission at 590 nm) using a FLUOstar
OPTIMA plate reader. The results were graphed to determine the
MICs. The lowest drug concentration effecting 100% growth inhibi-
tion was considered the MIC.
4.2.26. 5-Deoxy-5-hexadecylamino-bis-
5-deoxy-5-hexadecylamino-bis- -arabinitol (9k) was isolated
as a side-product during a strategy used to initially prepare 9i.
The protocol used was as follows: To a solution of -arabinose
D-arabinitol (9k)
D
D
(197 mg, 1.3 mmol) in EtOH (16.8 mL), was added hexadecylamine
(576 mg, 2.4 mmol), NaCNBH3 (124 mg, 2.0 mmol), and AcOH
(0.96 mL). The mixture was stirred at 50 °C for 18 h then the
solvent was removed under reduced pressure. The residue was
co-evaporated with 2ꢀ 25 mL of 5% HCl in methanol (w/v), then
purified by gradient flash silica chromatography (DCM/EtOH/
MeOH/35% NH3 (aq), 105/2/2/1 to 15/2/2/1, v/v/v/v) to give 9i as
a white solid (168.6 mg, 0.45 mmol, 34%) and5-deoxy-5-hexadecy-
Acknowledgments
This work was supported by the Lottery Health Board of New
Zealand, the Wellington Medical Research Foundation and the
Marsden Fund Council.
Supplementary data
lamino-bis-
(14 mg, 0.03 mmol, 4%). Rf = 0.17 (DCM/EtOH/MeOH/35% NH3
(aq), 5/2/2/1, v/v/v/v); [
26 = +9.0 (c 1.0, pyridine); IR (film)
3357, 2954, 2922, 2853, 1466, 1203, 1135, 1080 cmꢁ1
D-arabinitol (9k) also a white solid, as a by-product
Supplementary data (copies of 1H NMR and 13C NMR spectra of
all new compounds) associated with this article can be found, in
the online version, at http://dx.doi.org/10.1016/j.bmc.2015.12.036.
a]
D
.
1H NMR
(500 MHz, Pyridine-d5) d 4.98–4.89 (m, 1H, H-4), 4.59–4.54 (m,
1H, H-2), 4.52 (dd, J1a,2 = 3.8, J1a,1b = 10.7 Hz, 1H, H-1a), 4.39 (dd,
J1b,2 = 5.6, J1a,1b = 10.7 Hz, 1H, H-1b), 4.32 (d, J2,3 = 8.0 Hz, 1H,
H-3), 3.27 (dd, J4,5a = 7.9, J5a,5b = 12.9 Hz, 1H, H-5a), 3.12 (dd,
J4,5b = 5.4, J5a,5b = 12.9 Hz, 1H, H-5b), 2.78–2.65 (m, 2H, H-10),
1.63–1.54 (m, 2H, H-20), 1.32–1.15 (m, 26H, H-30-H-150), 0.88 (t, J
References and notes
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30.4, 30.3, 30.1, 30.0, 27.9, 23.3 (C30-C150), 26.6 (C20), 14.6 (C160);
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4.3.1. BCG Alamar Blue growth inhibition assay protocol
4.3.1.1. Bacteria.
per mL) were stored at ꢁ80 °C. These were defrosted with repeated
cycles of 10 s sonication, followed by 10 s on ice. A stock sample of
BCG was diluted to a concentration of 8 ꢀ 104 bugs per mL using
sterile Tween albumin broth [Dubos broth base/OADC] to suspend.
Stock solutions of BCG (1.5 ꢀ 107 bacteria
4.3.1.2. Compounds.
Compounds were dissolved in one of
two ways, depending on solubility: (a) for compounds soluble in
phosphate buffered saline (PBS) with up to 5% DMSO, a solution
was prepared with sterile PBS and sterile DMSO at double the
desired concentration and these were filter sterilised using Acro-
disc 13 mm syringe filters, with 0.22
eppendorf tubes. A 200 L aliquot of this solution was then added
12. Stocker, B. L.; Jongkees, S. A. K.; Win-Mason, A. L.; Dangerfield, E. M.; Withers,
S. G.; Timmer, M. S. M. Carbohydr. Res. 2013, 367, 29.
13. Win-Mason, A. L.; Jongkees, S. A. K.; Withers, S. G.; Tyler, P. C.; Timmer, M. S.
M.; Stocker, B. L. J. Org. Chem. 2011, 76, 9611.
14. Dangerfield, E. M.; Gulab, S. A.; Plunkett, C. H.; Timmer, M. S. M.; Stocker, B. L.
Carbohydr. Res. 2010, 345, 1360.
lm pore size into sterile
l
to the wells for serial dilution. (b) Compounds soluble only in
DMSO were prepared at forty times the desired starting concentra-
15. Dangerfield, E. M.; Plunkett, C. H.; Stocker, B. L.; Timmer, M. S. M. Molecules
2009, 14, 5298.
16. Yajko, D. M.; Madej, J. J.; Lancaster, M. V.; Sanders, C. A.; Cawthon, V. L.; Gee, B.;
Babst, A.; Hadley, W. K. J. Clin. Microbiol. 1995, 33, 2324.
17. Collins, L. A.; Franzblau, S. G. Antimicrob. Agents Chemother. 1997, 41, 1004.
18. For some examples of TB inhibitors based on AraT inhibitors, see: (a) Centrone,
C. A.; Lowary, T. L. J. Org. Chem. 2002, 67, 8862; (b) Naresh, K.; Bharati, B. K.;
Avaji, P. G.; Jayaraman, N.; Chatterji, D. Org. Biomol. Chem. 2010, 8, 592; (c)
Bosco, M.; Bisseret, P.; Constant, P.; Eustache, J. Tetrahedron Lett. 2007, 48, 153;
(d) Maddry, J. A.; Bansal, N.; Bermudez, L. E.; Comber, R. N.; Orme, I. M.; Suling,
W. J.; Wilson, L. N.; Reynolds, R. C. Bioorg. Med. Chem. Lett. 1998, 8, 237.
19. Protopopova, M.; Hanrahan, C.; Nikonenko, B.; Samala, R.; Chen, P.; Gearhart, J.;
Einck, L.; Nacy, C. A. J. Antimicrob. Chemother. 2005, 56, 968.
tion, in sterile eppendorf tubes. A 10
lL aliquot of this solution and
190 L of PBS was then added to the wells for serial dilution.
l
4.3.1.3. Minimum inhibitory concentration.
Optical bottom
black 96 well plates were used for the screening of the compounds.
The compounds were tested in triplicate and tested over three
serial dilutions, starting at a concentration of 500 mg/mL, with
repeated serial dilutions at a lower concentrations being per-
formed on compounds showing initial inhibitory activity. Ethamb-
utol was used as a positive control. To determine if the compounds
ˇ
20. Kovác, P.; Petríková, M. Carbohydr. Res. 1971, 19, 249.
Please cite this article in press as: Corkran, H. M.; et al. Bioorg. Med. Chem. (2016), http://dx.doi.org/10.1016/j.bmc.2015.12.036