Arabino mycolates from synthetic mycolic acids

Article abstract of DOI:10.1016/j.tet.2016.03.083

The synthesis of single mono-arabino mycolates, important lipid antigens from mycobacteria is described, using structurally defined synthetic mycolic acids. Preliminary assays indicate that these are differentially antigenic to antibodies in the serum of

Full text of DOI:10.1016/j.tet.2016.03.083

Accepted Manuscript
Arabino Mycolates from Synthetic Mycolic Acids
Mohsin O. Mohammed, Mark S. Baird, Juma’a R. Al Dulayymi, Alison Jones,
Christopher D. Gwenin
PII:
S0040-4020(16)30225-3
10.1016/j.tet.2016.03.083
TET 27623
DOI:
Reference:
To appear in: Tetrahedron
Received Date: 26 January 2016
Revised Date: 12 March 2016
Accepted Date: 24 March 2016
Please cite this article as: Mohammed MO, Baird MS, Al Dulayymi J’aR, Jones A, Gwenin CD, Arabino
Mycolates from Synthetic Mycolic Acids, Tetrahedron (2016), doi: 10.1016/j.tet.2016.03.083.
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ACCEPTED MANUSCRIPT
Graphical Abstract
Arabino Mycolates from Synthetic Mycolic Acids
Leave this area blank for abstract info.
Mohsin O. Mohammed, Mark S. Baird,*
Juma’a R. Al Dulayymi , Alison Jones and Christopher D. Gwenin
School of Chemistry, Bangor University, Bangor, Wales, LL57 2UW
R = synthetic mycolic acid

1
ACCEPTED MANUSCRIPT
Tetrahedron
journal homepage: www.elsevier.com
Arabino Mycolates from Synthetic Mycolic Acids
Mohsin O. Mohammed,a Mark S. Baird,* Juma’a R Al Dulayymi, Alison Jones and Christopher D. Gwenin
School of Chemistry, Bangor University, Bangor, Wales, LL57 2UW
ARTICLE INFO
ABSTRACT
The synthesis of single mono-arabino mycolates, important lipid antigens from mycobacteria is
described, using structurally defined synthetic mycolic acids. Preliminary assays indicate that
these are differentially antigenic to antibodies in the serum of people with active tuberculosis.
Article history:
Received
Received in revised form
Accepted
Available online
2009 Elsevier Ltd. All rights reserved
.
Keywords:
Mycobacteria
Tuberculosis
Arabino mycolates
Antigens
*Corresponding author. E-mail: chs028@bangor.ac.uk; fax: +44 1248 370528
a.
Current address: College of science, Chemistry department, Kirkuk University, Iraq

2
Tetrahedron
1. Introduction
of various mycobacteria under acidic conditions was reported.16-
22
ACCEPTED MANUSCRIPT
More recently, mass spectrometry and NMR have provided
powerful tools for the analysis of such molecules.23 Azuma and
The mycobacterial cell wall has a complex structure made up of
lipids, glycolipids, polysaccharides and proteins.1 There are four
major components: peptidoglycan (PG), mycolyl-arabinogalactan
(mAG), lipoarabinomannan (LAM) and extractable lipids.2 The
mycolyl-arabinogalactan (mAG) complex is the largest
component structure and acts as a permeability barrier that
prevents passage of antibiotics. It forms from cross bonding
Yamamura in 1962 isolated arabinomycolate as a D-arabinose-5-
mycolate and proved it was toxic to mice.17 Inflammatory
reactions similar to that observed after inoculation of live BCG
were induced in the lungs by trehalose monomycolate and
dimycolate (TMM and TDM) or glucose monomycolate (GMM)
isolated from BCG. However, the toxic reactions caused by
glycerol monomycolate (GroMM) and arabinose mono-mycolate
were characterized by an acute inflammatory process.24
between both
D-arabinofuranosyl (Araf) and-galactofuranosyl
(Galf ) with a long chain (C70-C90), α-alkyl branched β-
hydroxylated fatty acid, ‘mycolic acid’; carbohydrate (Araf) and
(Galf) are bound to peptidoglycan in the cell wall by an α-
rhamnopyranosyl-(1→3)-2-acetamido-2-deoxy-α-D-gluco-
L-
In 2006, the preparation of a tetramycolyl pentaarabinose using a
complex natural mixture of MAs was described.25 However, only
in 2010 were structural studies of the composition of the
arabinose mycolates of the cell wall of M. bovis reported. A two-
layer acid hydrolysis gave a number of fractions. One of these
was a penta-arabinose tetramycolate, one was an arabinose
mono-mycolate, while the others were hexa-arabinose, hepta-
arabinose and octa-arabinose tetramycolates. The mycolic acid
methyl esters released from each of these showed a mass
spectrometric pattern almost identical to the methyl esters
obtained by hydrolysis of the original cell wall. A comparison of
the penta-arabinose tetramycolate and arabinose monomycolate
with samples prepared by combining a mixture of natural MAs
with arabinose was reported.26 Ishiwata et al reported the
synthesis of a series of mono-, di- and tetra-arabinomycolates
found in the terminal position of cell wall skeleton of bacillus
Calmette Guerin from M. bovis, by using natural mycolic acid
mixtures extracted from the cell wall. They proved the biological
activity for synthesized compounds in a tumor necrosis factor
alpha (TNF-α) secretion-inducing assay; all the compounds
showed strong TNF-α inducing activity in vitro.25 The
mechanism of the activity of arabino-mycolate is not clear.22
Synthetic arabino-mycolates induce the production of TNF-α in
murine macrophage cell lines at an intensity similar to BCG-
CWS (cell wall skeleton). However the immunological activity
of natural arabino-mycolates isolated from BCG has not been
investigated, probably due to the complexity of the molecule.
Arabino-mycolates obtained by acid hydrolysis from CWS
(SMP-105) of M. bovis BCG Tokyo 172 strain consisted mainly
of mono-arabinose mono-mycolate, pentaarabinose tetra-
mycolate and hexa-arabinose tetramycolate fractions. Arabino-
mycolates significantly induced TNF-α production and enhanced
delayed type hypersensitivity (DTH) reactions against inactivated
tumour cells. Arabino-mycolates-induced TNF-α production was
completely dependent on TLR2 and MyD88 pathways. Thus
isolated natural arabino-mycolates possess potent adjuvant
immunostimulatory activity.27
pyranosyl phosphate disaccharide. The galactan part is linear and
composed of alternating β-(1→5) and β-(1→6) galactofuran
residues. Galactan and arabinan are bonded from C-5 of the β-
(1→6) link.3-6 Both galactosyl and arabinosyl units in mAG are
in a furanose form, less stable thermodynamically than the
pyranose form.7 It is believed that this plays a large role in raising
the flexibility of the polysaccharide and making the mycolic
acids (MAs) pack strongly by van de Waals interaction. Thereby,
the structure of the cell wall has extremely low permeability and
this provides the organism with high protection from drugs and
from its environment.1 Therefore, studying and understanding
mAG biosynthesis is an important strategy for developing new
anti-tuberculosis drugs. Indeed, isoniazid and ethambutol, two of
the standard antibiotics, target mAG biosynthesis; ethambutol
inhibits arabinosyltransferases which contribute to the
biosynthesis of the arabinan part of the polysaccharides while
isoniazid inhibits mycolic acid biosynthesis.8
It is approaching a century since Anderson began his ground
breaking studies of the lipid components of mycobacteria,
leading eventually to the characterization of MAs. These were
shown to be high-molecular weight hydroxy acids, with an
experimental formula of C88H172O4 or C88H176O4 which were very
hard to purify and not possible to crystallize.9,10 The long alkyl
branch on the α-position and hydroxyl groups in β-position in
MAs was proved by Asselineau in 1950.11 In brief, MAs can
defined as a complex group of long chain fatty α-alkyl β-alkyl
hydroxyl fatty acids (Figure 1).12,13
Intra-tumor injections of extracts of Re mutant Salmonella
typhimurium in combination with TDM or arabinose mycolate
were highly effective in producing regression of tumors in guinea
pigs. Similar extracts from M. bovis strain BCG and strain AN5
in combination with TDM also possessed tumor-regressive
activity. The activity was reduced when the arabinose mycolate
was substituted for the TDM. An extract of Coxiella burnetii, in
combination with either TDM or arabinose mycolate was also
active. Intracutaneous administration of Re glycolipid or aqueous
extracts from BCG in combination with trehalose or arabinose
mycolates did not produce life-threatening, clinical signs of
toxicity in young mice. If additional toxicity studies demonstrate
that adverse side effects can be satisfactorily controlled, these
water soluble extracts may prove beneficial in the treatment of
spontaneous tumors of humans and other animals.28.29 The
adjuvant activity of cell wall skeletons (mycolic acid-arabino-
galactan-mucopeptide) prepared from the cells of mycobacteria,
Figure 1: Generalised mycolic acid structure
The proximal group Y is generally a cis- alkene or cyclopropane,
or a trans-alkene or cyclopropane with a further methyl
substituent on the adjacent carbon distal from the hydroxyl acid.
The distal group X is normally a cis-cyclopropane (α-MAs), a –
(CHMeCHOMe)- (methoxy MAs) or a –(CHMeCO)- fragment
(keto MAs). Mixtures extracted from mycobacteria comprise
many individual MA containing a range of functional groups X
and Y (Figure 1) and but each also of several chain lengths.14
Anderson & Geiger in 1937 claimed the first extraction of
arabino-mycolate from the cell wall of Mycobacterium bovis
using natural organic solvent.15 Some fifty years ago, the
isolation of arabinose 5-mycolate by extraction of the cell walls

3
nocardia and corynebacteria was examined in vivo in mice and
guinea pigs. The cell wall skeletons of M. bovis BCG (BCG-
CWS), Nocardia asteroides 131 and Corynebacterium
diphtheriae PWC suspended in Freund's incomplete adjuvant
(FIA) as water-in-oil emulsions showed potent adjuvant activity
on the formation of circulating antibody and cell-mediated
immunity to bovine serum albumin (BSA), sheep erythrocytes
(SRBC) and sulfanylazo-bovine serum albumin (SA-BSA) in
mice and guinea pigs. After acetylation or acid treatment, BCG-
CWS retained its adjuvant activity, but the activity of BCG-CWS
was destroyed completely by alkaline treatment. The cell wall
constituents, arabinose-mycolate and arabino-galactan, prepared
activating the sugar as a tosylate raised the yield to 79%.
ACCEPTED MANUSCRIPT
Therefore, the hydroxyl group in the compound 4 was tosylated
by reaction with p-toluenesulfonyl chloride in dry pyridine and
catalytic 4-dimethylaminopyridine (DMAP) in dry CH2Cl2 at 0
ºC to afford the tosylate 5. Surprisingly, esters of arabinose with
simple fatty acids do not appear to have been reported. In a
model experiment, compound 4 was therefore first condensed
with behenic acid using DMAP and N,N-dicyclohexylcarbodi-
imide in dry CH2Cl2 to give compound 7a in 80% yield.36 This
was then hydrogenolysed in the presence of Pd(OH)2 and under a
hydrogen atmosphere to give the behenyl arabinose 8a.
Compound 5 was also reacted with palmitic acid by an alkylative
esterification using cesium hydrogen carbonate in dry DMF: THF
from BCG-CWS showed no adjuvant activity 30
.
at 70 C,25 to give compound 7b; this was then debenzylated to
0
Few studies of arabinose mycolates have been carried out and
little is known of the effect of structure on the
immunostimulatory activity of arabino-mycolates in activating
give the compound 8b. In the same way, compound 5 was
reacted with oleic acid to give the compound 7c; hydrogenolysis
in the presence of Pd(OH)2 under a hydrogen atmosphere gave
the corresponding stearyl arabinose, in which the double bond
was also saturated. However, deprotection with boron trichloride
in dichloromethane and methanol gave the oleoyl derivative 8c.
macrophages.6 However,
D
-arabinose-5-mycolate, purified from
bound lipids of the cell-wall skeleton of M. bovis BCG may be
prominent structure for recognition by host immunity 6
.
However, it is now clearly established that, in the case of free
MAs, the detailed structure controls their effects on a range of
cytokines and chemokines and that some lead to inflammatory
reactions on intratracheal instillation in mice, while others lead to
the formation of foam cells.31 We now report the synthesis of a
set of arabinose esters of stereochemically defined synthetic
MAs, as well as those of some simple fatty acids.
A series of synthetic MAs was then reacted with the compound 5
by alkylative esterification using cesium hydrogen carbonate as
in Scheme 2. Firstly, the methoxy cis-cyclopropane mycolic acid
6d,37 gave compound 7d, which was then debenzylated using
hydrogen and Pd(OH)2 catalyst to give the arabinose ester 8d.
This was achieved with no loss of the cyclopropane group, on the
basis of the proton and carbon NMR spectra. Although the
absolute stereochemistry of the cis-cyclopropane unit in MAs
remains unproven, this compound contains the stereochemistry
that appears likely, based on a common intermediate leading to
the different classes of mycolic acid.38 The 22 carbon length of
the α-chain is typical of mycobacteria such as Mycobacterium
kansasii.39 Condensation of methoxy-cis-cyclopropane mycolic
acid 6e,37 having a twenty two carbon chain in the α-position, and
the opposite absolute cis-cyclopropane stereochemistry, one
stereoisomer of which is present in M. kansasii, led to compound
7e. On hydrogenolysis as above, this gave compound 8e. In the
same way, esterification of methoxy-cis-cyclopropane mycolic
acid 6f,40 with a twenty four carbon chain at the α-position, a
stereoisomer of which is a major component of M. tuberculosis
methoxy-MA,39 but the opposite cyclopropane stereochemistry
to 6d gave compound 7f, which on hydrogenolysis led to
compound 8f.
2a Results and discussion: synthesis of arabinose mycolates
According to literature procedures, D-(-)-arabinose 1 was treated
with HCl (0.22 M) freshly prepared by addition of acetyl chloride
to anhydrous methanol at 0 ºC working up with pyridine rather
than ammonium carbonate,32 to give methyl-α,β-
D
-arabino
/β-
furanoside with predominant formation of the α-anomer (α-
D
D
3:2).33,34 In order to isolate these two anomers, in situ tritylation
of this mixture followed by column chromatography gave
methyl 5-O-trityl α-D-Araf 2 (45%) and 5-O-trityl β-D-Araf 3
(30%). Compound 2 was benzylated to protect the two secondary
hydroxyl groups by using benzyl bromide and sodium hydride in
dry DMF then the trityl group was removed from the primary
hydroxyl group by hydrolysis in 80% AcOH, affording
compound 4 (Scheme 1).25,34
The same procedure was repeated to prepare the arabino ester
compounds for the keto cis-cyclopropane mycolic acid 6g,37 with
a C22 alkyl chain in the α-position; this was used as the epimeric
mixture at the carbon adjacent to the ketone. This produced
compound 7g which on debenzylation gave the arabinose ester
8g. Ester 8g’, with the normal C24 M. tuberculosis α-chain length,
was also obtained from 6g’.41 In the same way the arabinose ester
8h, from the keto trans-cyclopropane mycolic acid 6h,42 again
epimeric adjacent to the ketone, was obtained in good yield.
Finally, the α-mycolic acid 6i,43,44 was esterified to give
compound 7i and then debenzylation to give 8i.
The yields for each of these coupling steps and deprotections are
given in Table 1. All the products were characterized by accurate
mass MALDI-MS, proton and carbon NMR and the spectra are
presented as Supplementary Information.
Scheme 1. Reagents and conditions: (i) HCl, CH3OH; (ii) Trityl chloride,
DMAP, pyridine, 0 ºC/R.T., then 70 ºC 4 h, 45% from α-anomer; (iii) a:
NaH, BnBr, DMF, 0 ºC/R.T. 2 h, b: 80% AcOH, 75 ºC for 4 h, 78%; (iv) 4-
toluenesulfonyl chloride, DMAP, pyridine, 90%.
According to the literature,25 direct esterification between the
primary hydroxyl group of Araf and a carboxyl group in natural
mycolic acid mixtures was achieved in a low yield (30%), while

4
Tetrahedron
ACCEPTED MANUSCRIPT
R
7 (%)
80
8 (%)
81
a
b
c
76
88
83
64
d
e
77
79
75
65
f
80
76
g
g’
h
80
63
71
66
92
80
R = stearate
i
87
80
R= CH3(CH2)20 a, CH3(CH2)14 b R =
c
Table 1: Yields for arabinose esters
2b. Assessment of arabinose mycolates as antigens
R=
R=
d
It is known that both trehalose esters of natural MAs, and the free
acids themselves are antigenic to antibodies in the serum of
patients infected with tuberculosis (TB).45,46 As an initial study of
the biological properties of the unique arabinose mycolates
prepared in this work, their antigenicity to 64 serum samples
taken from a Gambian population was determined using a
standard enzyme-linked immunosorbent assay (ELISA). The
samples, provided by the World Health Organisation from the
TDR TB Specimen Bank,47 were all from individuals with
symptoms of suspected tuberculosis. Nine were diagnosed as
having active TB using a range of clinical and biochemical
assays; 55 as were diagnosed as not having active TB. The
categorization was carried out using standard WHO protocols.
The assay used an IgG(Fc) conjugated secondary antibody to
detect the disease antibody – antigen interaction based on the
resulting absorbance at 492 nm. The results are summarized in
Table 2. This shows the median absorbance for active TB and no
active TB samples, the area under the curve (AUC) for a
Receiver Operating Characteristic (ROC) analysis of the data for
each antigen, the cut-off response set for a positive response for
each antigen in the assay, and the resulting sensitivity (% of
active TB diagnosed samples giving a response above the assay
cut-off) and specificity (% of no active TB samples giving a
response below the cut-off value).
e
R=
R=
f
g n = 21
g’ n= 23
R=
h
R=
i
Scheme 2: Synthesis of arabinose esters

5
Silica gel (Merck 7736) and silica gel plates used for column
ACCEPTED MANUSCRIPT
7d
7e
7f
7g
7h
7i
chromatography and thin layer chromatography were obtained
from Aldrich; separated components were detected using
variously UV light, I2 and phosphomolybdic acid solution in IMS
followed by charring. Infra-red (IR) spectra were carried out on
a Perkin-Elmer 1600 F.T.I.R. spectrometer as liquid films or KBr
disc (solid). Melting points were measured using a Gallenkamp
melting point apparatus. NMR spectra were carried out on a
Bruker AC250 or Advance 400 spectrometer. [α]D values were
recorded in CHCl3 on a POLAAR 2001 optical activity
polarimeter. Low resolution mass spectra were recorded on a
Bruker matrix-assisted laser desorption/ ionisation-time of flight
mass spectrometry (MALDI-TOF MS) values are given plus
sodium. Accurate mass measurements were carried out by the
EPSRC UK National Mass Spectrometry Facility at Swansea
University.
Median
active TB
0.64
0.40
0.78
0.50 0.48 1.18
Median no
active TB
0.39
.85
0.33
.69
0.46
.87
0.44 0.45 0.52
ROC AUC
.69
.67
.95
>.7
9
Cut-off for
‘active’ in
assay
>.35
9
>.27
9
>.45
9
>1.00 >.25
Active TB
samples above
the cut-off
1
2
9
3.1: Methyl 5-O-trityl-α-D-Araf 2
No active TB
samples above
the cut-off
Freshly prepared HCl solution in MeOH (resulting from mixing
acetyl chloride (2 mL) in MeOH (30 mL) at 0 ºC) was added to a
18
29
20
45
11
stirred solution of D-(-)-arabinose (5.0 g, 33 mmol) in anhydrous
MeOH (100 mL). Stirring was continued overnight at room
temperature, when a clear solution was obtained. The mixture
was neutralized by adding pyridine to pH 7 – 8, and the solid was
filtered and washed with MeOH (10 mL). The solvent was
evaporated to give a residue which was purified by column
chromatography eluting with chloroform: acetone (3:5) to give a
Sensitivity
Specificity
100
100
100
64
100
20
100
18
100
80
67
47
Table 2. Responses for a set of 64 human serum samples from
Gambia (9 diagnosed with active TB, 55 with no active TB) to
synthetic arabinose mycolates coated as antigens on ELISA
plates, using IgG(Fc) secondary antibody conjugate to determine
the binding. The closer the ROC AUC is to 1, the higher the
predictive value of the assay.
colourless oil, methyl–α,β-
(6.42 g, 23.0 mmol) and 4-(dimethylamino)pyridine (2.57 g, 21.0
mmol) were added to a stirred solution of methyl-α,β- -Araf
D-Araf (4.3 g, 78%). Trityl chloride
D
(3.12 g, 19.0 mmol) in anhydrous pyridine (60 mL) and the
mixture was stirred at room temperature overnight then heated on
oil bath at 70 ºC for 4 h. The mixture was cooled to room
temperature and poured into ice/water (300 mL). The organic
phase was separated and the aqueous phase was extracted with
ethyl acetate (3 × 100 mL). The combined organic layers were
washed with aq. NaHCO3 solution (5%, 100 mL), dried and the
solvent was evaporated. The residue was purified by column
chromatography eluting started from 10% to 50% hexane/ethyl
acetate gave the title compound as a colorless oil 2 (3.3 g, 45%)
This shows that the six synthetic arabinose mycolates are
detected to a different degree by antibodies in the serum of
patients infected with active tuberculosis; in some cases the
response is essentially no different between active TB and no
active TB samples, whereas in others, such as 7i, the assay result
correlates well with the clinical diagnosis. The active TB status
had been determined using a range of standard assays employed
by the WHO, including being smear and culture positive; the no
active TB samples were all from patients showing some of the
symptoms of TB, but diagnosed using the same methods as not
having active TB, and being both smear and culture negative.
Although this is a very small sample set, it is interesting that the
highest distinction between active TB and no active TB sets is
given with the arabinose mycolates 7d and 7i which probably
represent the natural stereochemistries of methoxy- and α-
mycolates. Work is on-going to validate these results on a larger
sample set.
and 5–O-trityl-β-D-Araf 3 (2.3 g, 30%). The mixture of anomers
showed δH (400 MHz, CDCl3), δC (125 MHz, CDCl3) and vmax
identical to the literature.33.34
3.2: Methyl 2,3-di-O-benzyl-α-D-Araf 4
A solution of 2 (3.20 g, 7.88 mmol) in dry DMF (80 mL) was
added dropwise to a stirred suspension of NaH (0.85 g) (60%
w/w, dispersion in mineral oil, washed with petrol three times) at
room temperature under nitrogen. The mixture was stirred for 30
min. then benzyl bromide (2.5 mL, 3.6 g, 21.0 mmol) in dry
DMF (50 mL) was added. The mixture was stirred at room
temperature for 20 h and then quenched by slow addition of
water (15 mL) and diluted with ether (25 mL). The organic layer
was separated and the aqueous layer was extracted with ether (2
× 100 mL). The combined extracts were washed with water (50
mL), brine (50 mL), dried and evaporated under reduced
pressure. Aqueous acetic acid (80%, 30 mL) was added to the
crude product and the mixture was stirred and heated at 75 ºC for
4 h, then diluted with water (20 mL) and ether (20 mL) and the
organic layer was separated and the aqueous layer extracted with
ether (2 × 100 mL). The combined organic layers were washed
with water (50 mL), sat. aq. NaHCO3 (50 mL) and brine (50 mL),
dried and the solvent was evaporated. Column chromatography
eluting with petrol/ethyl acetate (7:3) gave the title compound as
The effects of selected examples of the above synthetic arabinose
mono-mycolates on the Mincle receptor in M. tuberculosis will
be described elsewhere.48
3. Experimental section
Chemicals used were obtained from commercial suppliers
(Sigma, Aldrich, and Alfa Aeser) or prepared from them by the
methods described. Solvents which were required to be dry, e.g.
ether, tetrahydrofuran were dried over sodium wire and
benzophenone under nitrogen, while dichloromethane and
HMPA were dried over calcium hydride. All reagents and
solvents used were of reagent grade unless otherwise stated.
Organic solutions were dried over anhydrous magnesium sulfate.

6
Tetrahedron
ACCEPTED MANUSCRIPT
a colorless oil 4 (2.12 g, 78%) [MALDI-Found (M+Na)+:
mixture was stirred for 24 h then filtered and the precipitate
367.3; C20H24NaO5, requires: 367.1], [α] 22 +89 (c 0.10, CHCl3)
washed with CH2Cl2 (10 mL), the filtrate was evaporated and the
residue was purified by column chromatography eluting with
hexane/ethyl acetate (1:1) to afford the title compound as a
colorless oil 8a (0.0079 g, 81%) [MALDI-Found (M+Na)+:
509.5, C28H54NaO6 requires: 509.3], [α] 2D2 +46 (c 0.10, CHCl3),
[lit.26 [α] 24 +83.2 (c 1.14, CHCl3)], whichDshowed δH (400 MHz,
D
CDCl3): 7.40 – 7.28 (10H, m), 4.95 (1H, s), 4.61 (1H, d, J 12.0
Hz), 4.54 (1H, d, J 12 Hz), 4.53 (1H, d, J 12 Hz), 4.50 (1H, d, J
12 Hz), 4.18 – 4.12 (1H, m), 4.02 – 3.96 (2H, m), 3.85 (1H, dd, J
12.1, 2.8 Hz), 3.65 (1H, dd, J 12.1, 4.1 Hz), 3.40 (3H, s), 2.93
(1H, br. s); δC (101 MHz, CDCl3): 137.6, 137.2, 128.4, 128.4,
127.9, 127.8, 127.79, 127.7, 107.3, 87.6, 82.5, 82.2, 72.3, 71.8,
62.1, 54.8; νmax: 3466 br., 3089, 3064, 3031, 2925, 1725, 1605,
1454, 739 cm-1. All data were identical to the literature.25,34
which showed δH (500 MHz, CDCl3): 4.92 (1H, s), 4.32 – 4.29
(2H, m), 4.22 (1H, dd, J 7.1, 3.9 Hz), 4.09 (1H, br. s), 3.90 (1H,
br.s), 3.42 (3H, s), 2.81 – 2.67 (1H, m), 2.51 – 2.39 (1H, m), 2.38
– 2.32 (2H, m), 1.62 (2H, dd, J 14.7, 7.3 Hz), 1.35 – 1.20 (36H,
m), 0.89 (3H, t, J 7.0 Hz); δC (101 MHz, CDCl3): 173.5, 108.8,
83.7, 79.8, 78.0, 63.8, 55.0, 34.1, 31.9, 29.7, 29.6, 29.5, 29.4,
29.3, 29.2, 29.0, 24.8, 22.6, 14.1 ; νmax: br. 3500, 2917,
2850,1739, 1464, 1100,758 cm-1.
3.3: Methyl 2,3-di-O-benzyl-5-O-p-toluensulfonyl-α-D-Araf 5
p-Toluenesulfonyl chloride (0.60 g, 3.14 mmol) was added to a
stirred solution of 4 (0.50 g, 1.45 mmol), pyridine (0.96 g,
0.98mL, 12.4 mmol) and DMAP (0.10g, 0.81 mmol) in dry
CH2Cl2 (5 mL) at 0 ºC. The mixture was stirred at room
temperature for 16 h then diluted with ethyl acetate (10 mL), the
organic layer was separated and the aqueous layer was re-
extracted with ethyl acetate (2 × 100 mL). The combined organic
layer were washed with water (50 mL), brine (50 mL), dried and
the solvent was evaporated. The residue was purified by column
chromatography eluting with hexane/ethyl acetate (4:1) affording
the title compound as a colorless oil 5 (0.65 g, 90%), [α] 2D3 +55
3.5: Methyl 2,3-di-O-benzyl-5-O-palmitoyl-α-D-Araf 8b
(a) Cesium hydrogen carbonate (0.19 g, 1.0 mmol) was added to
a stirred solution of 5 (0.1 g, 0.2 mmol) and palmitic acid (0.061
g, 0.237 mmol) in dry DMF:THF (1: 5, 5 mL) at room
temperature and the reaction mixture was stirred at 70 ºC for two
days. The suspension was diluted with ethyl acetate (10 mL) and
water (10 mL). The organic layer was separated and the aqueous
layer was re-extracted with ethyl acetate (2 × 10 mL). The
combined organic layers were washed with water (15 mL) and
brine (15 mL), dried, filtered and evaporated to give a thick oil
residue. The residue was purified by column chromatography
eluting with hexane/ethyl acetate (10:1) to give methyl 2,3-di-O-
(c 0.10, CHCl3) [lit.25 [α] 2D7 +57.0 (c 0.40, CHCl3)], which
benzyl-5-O-palmitoyl-α-D
-Araf 7b (0.089 g, 76%).25 as a
colorless oil [MALDI-Found (M+Na)+ : 605.1, C36H54NaO6
requires: 605.3], [α] 2D2 +37 (c 0.10, CHCl3), which showed δH
showed δH (400 MHz, CDCl3): 7.77 (2H, d, J 8.2 Hz), 7.41 –
7.25 (12H, m), 4.85 (1H, s), 4.56 – 4.38 (4H, m), 4.22 – 4.15
(1H, m), 4.12 (2H, br d, J 4.6 Hz), 3.94 (1H, br d, J 2.6 Hz), 3.81
(1H, dd, J 5.9, 2.7 Hz), 3.33 (3H, s), 2.42 (3H, s); δC (101 MHz,
CDCl3): 144.8, 137.3, 137.2, 132.7, 129.7, 128.4, 128.0, 127.9,
127.89, 127.8, 127.79, 107.4, 87.5, 82.8, 79.2, 72.2, 71.9, 68.8,
55.1, 21.6; νmax: 3064, 3032, 2916, 1741, 1454, 1365, 1177, 698
cm-1. All data were identical to the literature.25
(500 MHz, CDCl3): 7.39 – 7.28 (10H, m), 4.95 (1H, s), 4.59 (1H,
d, J 12.0 Hz), 4.57 (1H, d, J 11.7 Hz), 4.52 (1H, d, J 12.0 Hz),
4.49 (1H, d, J 11.8 Hz), 4.29 (1H, dd, J 11.3, 2.9 Hz), 4.24 – 4.15
(2H, m), 4.01 (1H, dd, J 3.0, 1.2 Hz), 3.84 (1H, dd, J 6.2, 2.9
Hz), 3.40 (3H, s), 2.32 – 2.27 (2H, m), 1.64 – 1.54 (2H, m), 1.26
(24H, s), 0.89 (3H, t, J 7.0 Hz); δC (101 MHz, CDCl3): 173.6,
137.5, 137.3, 128.4, 128.3, 127.9, 127.8, 107.3, 88.0, 83.4, 79.3,
72.3, 72.0, 63.5, 54.9, 34.1, 31.9, 29.7, 29.68, 29.6, 29.4, 29.32,
29.3, 29.1, 24.8, 22.7, 14.1; νmax: 3064, 3032, 2924, 2853, 1740,
1455, 1365, 1107, 735 cm-1 .
3.4: Methyl 5-O-behenoyl-α-D-Araf 8a
(a) A solution of N,N`-dicyclohexylcarbodimide (0.089 g, 0.432
mmol) in dry CH2Cl2 (1 mL) was added dropwise to a stirred
solution of 4 (0.10g, 0.29 mmol), DMAP (0.042 g, 0.343
mmol) and behenic acid (0.10 g, 0.29 mmol) in dry CH2Cl2 (1
mL) at 0 ºC under nitrogen. The mixture was stirred for 30 min.
then the precipitate of dicyclohexyl urea was filtered off and
washed with CH2Cl2 (10 mL), the solvent was evaporated and
the residue was purified by column chromatography eluting
with hexane/ethyl acetate (10:1) to afford methyl 2,3-di-O-
(b) Palladium hydroxide on activated charcoal (20% Pd(OH)2-C ,
0.01 g, 0.15 fold by weight ) was added to a stirred solution of
7b (0.06 g, 0.10 mmol) in dry CH2Cl2 : MeOH, (1:1, 2 mL) at
room temperature under hydrogen atmosphere. The reaction
mixture was stirred for 24 h then filtered and the precipitate was
washed with CH2Cl2 (10 mL), the filtrate was evaporated and the
residue was purified by column chromatography eluting with
hexane/ethyl acetate (1:1) to afford the title compound as a
colorless oil 8b (0.04 g, 88%),34 [α]1D8 +1.2 (c 0.10, CHCl3)
[MALDI-Found (M+Na)+: 425.5, C22H42NaO6 requires: 425.2]
which showed δH (500 MHz, CDCl3): 4.93 (1H, s), 4.33 – 4.30
(2H, m), 4.22 (1H, dd, J 6.9, 3.9 Hz), 4.09 (1H, br. s), 3.91 (1H,
br. s), 3.43 (3H, s), 2.35 (2H, t, J 7.6 Hz), 1.67 – 1.60 (2H, m),
1.56 (2H, br. s), 1.36 – 1.21 (24H, m), 0.89 (3H, t, J 6.9 Hz); δC
(101 MHz, CDCl3): 173.5, 108.8, 83.8, 79.8, 78.0, 63.8, 55.1,
34.1, 31.9, 29.6, 29.6, 29.5, 29.4, 29.3, 29.2, 29.0, 24.8, 22.6,
14.1 ; νmax: br. 3308, 2916, 2848, 1742, 1472, 1099, 728 cm-1.
benzyl-5-O-behenoyl-α-D
-Araf31 as a colorless oil 7a (0.15 g,
80%) [MALDI-Found (M+Na)+: 689.5, C42H66NaO6 requires:
689.4], [α] 2D4 +40 (c 0.10, CHCl3), which showed δH (500
MHz, CDCl3): 7.41 – 7.28 (10H, m), 4.95 (1H, s), 4.59 (1H, d, J
12.0 Hz), 4.57 (1H, d, J 12 Hz), 4.52 (1H, d, J 12 Hz), 4.49 (1H,
d, J 12 Hz), 4.29 (1H, dd, J 11.3, 2.8 Hz), 4.24 – 4.15 (2H, m),
4.01 (1H, dd, J 2.9, 1.1 Hz), 3.84 (1H, dd, J 6.3, 2.9 Hz), 3.40
(3H, s), 2.33 – 2.26 (2H, m), 1.64 – 1.52 (2H, m), 1.34 – 1.18
(36H, m), 0.89 (3H, t, J 6.9 Hz); δC (101 MHz, CDCl3): 173.6,
137.5, 137.4, 128.45, 128.43, 127.9, 127.8, 107.3, 88.0, 83.4,
79.4, 72.3, 72.0, 63.5, 55.0, 34.1, 31.9, 29.7, 29.6, 29.62, 29.5,
29.3, 29.27, 29.1, 24.8, 22.7, 14.1; νmax: 3034, 2915, 2849,
1741, 1471, 1100, 758 cm-1.
3.6: Methyl 5-O-oleoyl-α-D-Araf 8c
(a) Cesium hydrogen carbonate (0.47 g, 2.42 mmol) was added to
a stirred solution of 5 (0.26 g, 0.52 mmol) and oleic acid (0.10 g,
0.35 mmol) in dry DMF:THF (1:5, 2 mL) at room temperature
and the mixture was stirred at 70 ºC for two days. The suspension
was diluted with ethyl acetate (10 mL) and water (10 mL). The
(b) Palladium hydroxide on activated charcoal (20% Pd(OH)2-C,
0.0026 g, 0.15 fold by weight) was added to a stirred solution of
7a (0.010g, 0.01 mmol) in dry CH2Cl2: MeOH (1:1, 2 mL) at
room temperature under hydrogen atmosphere. The reaction

7
organic layer was separated and the aqueous layer was re-
extracted with ethyl acetate (2 × 10 mL). The combined organic
layers were washed with water (15 mL) and brine (15 mL). The
organic layer was dried, filtered and evaporated to give a thick oil
residue. This was purified by column chromatography eluting
with hexane/ethyl acetate (7:2) to afford methyl 2,3-di-O-benzyl-
5-O-oleoyl-α-D-Araf as a colorless oil 7c (0.18 g, 83%)
[MALDI-Found (M+Na)+ : 631.3, C38H56NaO6 requires : 631.4],
[α] 2D2 +45 (c 0.10, CHCl3) which showed δH (400 MHz, CDCl3):
(a) Cesium hydrogen carbonate (0.056 g, 0.288 mmol) was
ACCEPTED MANUSCRIPT
added to a solution of 5 (0.031g, 0.062 mmol) and (R)-2-{(R)-1-
hydroxy-18-[(1S,2R)-2-((17S,18S)-17-methoxy-18-
methylhexatriacontyl) cyclopropyl]octadecyl}tetracosanoic acid
(0.051g, 0.041 mmol)37 in dry DMF:THF (1:5, 2 mL) at room
temperature and the mixture was stirred at 70 ºC for two days.
Work up as before gave methyl 2,3-di-O-benzyl-5-O-(2-{(R)-1-
hydroxy-18-[(1S,2R)-2-[(17S,18S
)-17-methoxy-18-
methylhexatriacontyl]cyclopropyl]octadecyl}- tetracosanoate) α-
D
- Araf 7d as a thick colorless oil (0.05 g, 77%), [Found
7.39 – 7.28 (10H, m), 5.41 – 5.30 (2H, m), 4.95 (1H, s), 4.59
(1H, d, J 12.0 Hz), 4.57 (1H, d, J 12 Hz), 4.52 (1H, d, J 12 Hz),
4.49 (1H, d, J 12 Hz), 4.29 (1H, dd, J 11.0, 2.5 Hz), 4.24 – 4.15
(2H, m), 4.0 (1H, br d, J 2.1 Hz), 3.84 (1H, dd, J 6.2, 2.8 Hz),
3.40 (3H, s), 2.30 (2H, t, J 7.6 Hz), 2.06 – 1.97 (4H, m), 1.64 –
1.54 (4H, m), 1.28 (18H, br m), 0.89 (3H, t, J 6.8 Hz); δC (101
MHz, CDCl3): 173.6, 137.5, 137.4, 130.0, 129.7, 128.5, 128.4,
127.9, 107.3, 87.9, 83.3, 79.3, 72.3, 72.0, 63.5, 55.0, 34.0, 31.9,
29.7, 29.6, 29.5, 29.3, 29.17, 29.11, 29.1, 27.25, 27.2, 24.8, 22.6,
14.1; νmax: 3005, 3089, 3031, 2926, 2855, 1740, 1454, 1050, 735
cm-1.
(M+Na)+:1574.4044, C103H186NaO8 requires: 1574.4040]; [α] 2D0
+23 (c 0.10, CHCl3); δH (400 MHz, CDCl3): 7.39 – 7.28 (10H,
m), 4.92 (1H, s), 4.58 (1H, d, J 12.0 Hz), 4.56 (1H, d, J 12 Hz),
4.51 (1H, d, J 12 Hz), 4.48 (1H, d, J 12 Hz), 4.33 – 4.27 (2H, m),
4.25 – 4.19 (1H, m), 3.99 (1H, br d, J 2.0 Hz), 3.84 (1H, dd, J
6.4, 2.6 Hz), 3.67 – 3.59 (1H, m), 3.37 (3H, s), 3.35 (3H, s), 2.99
– 2.94 (1H, m), 2.52 (1H, d, J 8.3 Hz), 2.43 (1H, dt, J 9.4, 5.4
Hz), 1.72 – 1.61 (2H, m), 1.59 – 1.03 (141H, m), 0.89 (6H, t, J
6.8 Hz), 0.86 (3H, d, J 6.9 Hz), 0.70 – 0.61 (2H, m), 0.60 – 0.53
(1H, dt, J 4, 8 Hz), –0.33 (1H, br. q J 5.1 Hz); δC (101 MHz,
CDCl3): 175.0, 128.5, 128.4, 127.9, 127.8, 107.2, 87.9, 85.4,
83.7, 79.4, 72.4, 72.1, 63.5, 57.7, 54.9, 51.53, 35.52, 35.3, 32.36,
31.9, 30.5, 30.48, 30.4, 30.3, 30.23, 30.12, 30.0, 29.98, 29.94,
29.9, 29.86, 29.84, 29.7, 29.6, 29.5, 29.4, 29.3, 29.3, 29.26, 29.2,
29.19, 29.1, 29.0, 29.04, 28.7, 28.67, 27.5, 27.4, 26.1, 25.7, 22.7,
22.6, 15.7, 14.8, 14.1, 10.9; νmax: 3479, 3064, 2924, 2853, 1735,
1494, 1455, 1100 cm-1.
(b) Boron trichloride49 (0.32 mL, 1M) was added to a stirred
solution of compound (7c) (0.02 g, 0.03 mmol) in dry CH2Cl2 (2
mL) at -78 ºC. The mixture was stirred for 2 h then quenched
with CH2Cl2:MeOH (1:1, 2 mL) and the solvent was evaporated
and the residue was purified by column chromatography eluting
with hexane/ethyl acetate (1:1) to afford the title compound (7c)
(9 mg, 64%) [Found (MALDI) (M+Na)+: 451.3, C24H44NaO6
requires: 451.3], [α]1D8 +5.0 (c 0.10, CHCl3) which showed δH
(b) Palladium hydroxide on activated charcoal (20% Pd(OH)2-C
, 0.0033 g , 0.15 fold by weight) was added to a stirred solution
of 7d (0.022 g, 0.014 mmol) in dry CH2Cl2 : MeOH (1:1, 2 mL)
at room temperature under hydrogen atmosphere. The mixture
was stirred overnight; work up as before gave the title compound
as a thick colorless oil 8d (0.016 g, 79%) [Found (M+NH4)+:
1389.3550, C89H178NO8 requires: 1389.3552]; [α] 2D1 +28 (c
(400 MHz, CDCl3 + few drops CD3OD): 5.35 – 5.26 (2H, m),
4.77 (1H, d, J 4.3 Hz), 4.24 (1H, dd, J 11.7, 2.1 Hz), 4.11 – 4.05
(1H, m), 4.02 – 3.97 (1H, m), 3.97 – 3.91 (2H, m), 3.39 (3H, s),
2.32 (2H, t, J 7.6 Hz), 2.04 – 1.92 (4H, m), 1.65 – 1.54 (2H, m),
1.35 – 1.15 (22H, m), 0.84 (3H, t, J 6.7 Hz); δC (101 MHz,
CDCl3 + few drops CD3OD): 174.3, 129.96, 129.7, 102.3, 79.9,
77.7, 75.8, 65.4, 55.2, 49.6, 49.3, 49.1, 48.9, 48.7, 34.1, 31.8,
29.7, 29.6, 29.4, 29.2, 29.1, 29.0, 27.1, 27.09, 24.8, 22.6, 13.98;
νmax: br. 3468, 2917, 2850, 1735, 1454, 1050, 824 cm-1.
0.10, CHCl3); δH (400 MHz, CDCl3): 4.89 (1H, s), 4.49 (1H, dd,
J 12, 4.0 Hz), 4.35 (1H, dd, J 12, 4.1 Hz), 4.21 – 4.15 (1H, m),
4.07 (1H, br.s), 3.98 (1H, br.s), 3.77 – 3.66 (1H, m), 3.41 (3H, s),
3.35 (3 H, s), 3.01 – 2.92 (1H, m), 2.85 (1H, s), 2.54 – 2.36 (1H,
ddd, J 10.1, 6.9, 5.1 Hz), 1.48 – 1.17 (144H, m), 0.89 (6H, t, J
6.8 Hz), 0.86 (3H, d, J 6.9 Hz), 0.70 – 0.61 (2H, m), 0.61 – 0.52
(1H, dt, J 7.9, 3.9 Hz), -0.33 (1H, br. q J 5.1 Hz); δC (101 MHz,
CDCl3): 174.9, 108.7, 85.4, 83.6, 80.5, 78.4, 72.8, 63.2, 57.7,
54.9, 52.3, 35.3, 35.2, 32.3, 31.9, 30.7, 30.69, 30.64, 30.62,
30.55, 30.5, 30.4, 30.37, 30.3, 30.28, 30.2, 30.1, 30.0, 29.9, 29.8,
29.7, 29.6, 29.57, 29.55, 29.5, 29.4, 29.3, 29.28, 29.25, 29.2,
29.1, 29.0, 29.03, 29.0, 28.9, 28.7, 28.6, 28.5, 28.3, 27.5, 27.4,
26.1, 25.4, 22.6, 15.7, 14.8, 14.1, 10.9; νmax: br.3435, 2918, 2850,
1732, 1455,1100 cm-1.
3.7: Methyl 5-O-stearyl-α-D-Araf
Palladium hydroxide on activated charcoal (20% Pd(OH)2-C,
0.012 g, 0.15 fold by weight) was added to a stirred solution of
7c (0.08 g, 0.13 mmol) in dry (CH2Cl2 : MeOH, 1:1, 3 mL) at
room temperature under hydrogen atmosphere. The mixture was
stirred for 24 h then filtered and the precipitate washed with
CH2Cl2 (15 mL), the filtrate was evaporated and the residue was
purified by column chromatography eluting with hexane / ethyl
acetate (1:1) to afford the title compound as a colorless oil
(0.045 g, 80%) [MALDI-Found (M+Na)+: 453.0, C24H46NaO6
requires: 453.3]; [α]1D8 +2 (c 0.1, CHCl3); δH (400 MHz, CDCl3):
4.92 (1H, s), 4.30 (2H, d, J 4.0 Hz), 4.21 (1H, dd, J 7.3, 3.8 Hz),
4.17 – 4.06 (1H, m), 3.90 (1H, br d, J 10.1 Hz), 3.42 (3H, s), 2.77
(1H, d, J 10.2 Hz), 2.52 (1H, d, J 7.9 Hz), 2.35 (2H, t, J 7.6 Hz),
1.69 – 1.58 (2H, m), 1.34 – 1.21 (28H, m), 0.88 (3H, t, J 6.8 Hz);
δC (101 MHz, CDCl3): 173.5, 108.8, 83.7, 79.8, 78.0, 63.8, 55.1,
50.8, 34.1, 31.9, 29.7, 29.6, 29.6, 29.4, 29.3, 29.2, 29.05, 24.79,
22.67, 14.10; νmax: br. 3468, 2917, 2850, 1735, 1454, 1050, 824
cm-1.
3.9: Methyl 5-O-(2-[(R)-1-hydroxy-18-[(1R ,2S)-2-[(17S,18S)-
17-methoxy-18-methylhexatriacontyl]cyclo propyl]octadecyl-
]tetracosanoate) α-D-Araf 8e
(a) Cesium hydrogen carbonate (0.098 g, 0.505 mmol) was
added to a stirred solution of 5 (0.0546 g, 0.1000 mmol) and (R)-
2-{(R)-1-hydroxy-18-[(1R,2S)-2-((17S,18S)-17-methoxy-18-
methyl-hexatriacontyl)cyclopropyl]octadecyl}tetracosanoic acid
(0.100 g, 0.081 mmol)37 in dry DMF:THF (1:5, 2 mL) at room
temperature and the reaction mixture was stirred at 70 ºC for two
days. The suspension was diluted with ethyl acetate (10 mL) and
water (10 mL). The organic layer was separated and the aqueous
layer was re-extracted with ethyl acetate (2 × 10 mL). The
combined organic layers were washed with water (15 mL) and
brine (15 mL), then dried, filtered and evaporated to give a thick
3.8: Methyl 5-O-(2-{(R)-1-hydroxy-18-[(1S, 2R)-2-[(17S, 18S)
-17-methoxy-18-methylhexatriacontyl]cyclopropyl]octadecyl}
tetracosanoate) α-D-Araf 8d

8
Tetrahedron
ACCEPTED MANUSCRIPT
oil residue. The residue was purified by column
3.35 (3H, s), 3.00 – 2.93 (1H, m), 2.50 (1H, d, J 8.2 Hz), 2.44
chromatography eluting with hexane/ethyl acetate (10:1) to give
methyl 2,3-di-O-benzyl-5-O-(2-[(R)-1-hydroxy-18-[(1R,2S)-2-
[(17S,18S)-17-methoxy-18-methyl-hexatriacontyl]-
cyclopropyl]octadecyl]tetracosanoate)α-D-Araf as a colorless
thick oil 7e (0.095 g, 75%) [Found (M+Na)+: 1574.4085,
C103H186NaO8 requires: 1574.4040], [α] 2D3 +18 (c 0.10, CHCl3),
(1H, dt, J 8.7, 5.4 Hz), 1.75 – 1.56 (2H, m), 1.58 – 1.04 (145H,
m), 0.89 (6H, t, J 6.8 Hz), 0.86 (3H, d, J 6.9 Hz), 0.70 – 0.62
(2H, m), 0.57 (1H, dt, J 8 4 Hz), -0.32 (1H, br. q J 5.0 Hz); δC
(101 MHz, CDCl3): 175.0, 137.4, 137.3, 128.5, 128.4, 128.0,
127.9, 127.89, 127.88, 127.87, 127.86, 107.2, 87.8, 85.4, 83.7,
79.4, 72.4, 72.2, 72.1, 63.4, 57.7, 54.9, 51.5, 35.5, 35.3, 32.4,
31.9, 30.8, 30.7, 30.67, 30.6, 30.55, 30.5, 30.45, 30.4, 30.38,
30.37, 30.36, 30.2, 30.17, 30.16, 30.15, 30.0, 29.9, 29.8, 29.7,
29.6, 29.5, 29.4, 29.3, 29.27, 29.26, 29.2, 29.14, 29.1, 29.07,
29.02, 29.01, 28.94, 28.9, 28.7, 28.65, 28.6, 28.52, 28.5, 27.5,
27.4, 26.1, 25.7, 22.6, 15.7, 14.8, 14.1, 10.9 ; νmax: br. 3522,
3064, 2921, 2851 , 1723, 1467, 1027, 732 cm-1.
which showed δH (500 MHz, CDCl3): 7.38 – 7.29 (10H, m), 4.92
(1H, s), 4.58 (1H, d, J 12 Hz), 4.56 (1H, d, J 12 Hz), 4.51 (1H, d,
J 12 Hz), 4.48 (1H, d, J 12 Hz), 4.31 – 4.28 (2H, m), 4.24 – 4.20
(1H, m), 3.99 (1H, dd, J 2.7, 0.9 Hz), 3.84 (1H, dd, J 6.4, 2.6
Hz), 3.66 – 3.60 (1H, m), 3.38 (3H, s), 3.35 (3H, s), 3.00 – 2.93
(1H, m), 2.52 (1H, br. s), 2.43 (1H, dt, J 9.1, 5.5 Hz), 1.72 – 1.61
(2H, m), 1.60 – 1.07 (141H, m), 0.89 (6H, t, J 6.9 Hz), 0.86 (3H,
d, J 6.8 Hz), 0.69 – 0.62 (2H, m), 0.57 (1H, dt, J 8.4, 4.1 Hz), –
0.32 (1H, q, J 5.2 Hz); δC (101 MHz, CDCl3): 175.0, 137.5,
137.3, 128.6, 128.5, 127.9, 127.8, 107.2, 87.9, 85.4, 83.7, 79.4,
72.4, 72.2, 72.1, 63.5, 57.7, 54.9, 51.5, 35.5, 35.3, 32.5, 32.3,
31.9, 31.3, 31.0, 30.9, 30.8, 30.79, 30.6, 30.58, 30.5, 30.47, 30.4,
30.38, 30.37, 30.3, 30.29, 30.2, 30.19, 30.1, 29.9, 29.8, 29.7,
29.6, 29.5, 29.4, 29.3, 29.2, 29.18, 29.17, 29.16, 29.15, 29.0,
28.9, 28.8, 28.7, 28.5, 28.4, 28.3, 27.6, 27.4, 27.3, 26.9, 26.1,
25.7, 22.6, 15.7, 14.8, 14.1, 10.9 ; νmax : 3479, 3064, 2923, 2853,
1733, 1465, 1100, 721 cm-1.
(b) Palladium hydroxide on activated charcoal (20% Pd(OH)2-C ,
0.003 g , 0.15 fold by weight) was added to a stirred solution of
7e (0.020 g, 0.012 mmol) in dry CH2Cl2 : MeOH (1:1, 2 mL) at
room temperature under hydrogen atmosphere. The mixture was
stirred overnight then filtered and the solvent was evaporated to
give a residue; column chromatography eluting with hexane/ethyl
acetate (1:1) gave the title compound as a colorless oil 8e (0.011
g, 65%) [Found (M+Na)+: 1394.3138, C89H174NaO8 requires:
1394.3101], [α]1D6 +10 (c 0.70, CHCl3); δH (400 MHz, CDCl3):
(b) Palladium hydroxide on activated charcoal (20% Pd(OH)2-C,
0.003 g, 0.15 fold by weight) was added to a stirred solution of 7f
(0.020 g, 0.012 mmol) in dry CH2Cl2 : MeOH (1:1, 2 mL) at
room temperature under hydrogen atmosphere. The mixture was
stirred overnight then worked up as before to give the title
compound as a thick colorless oil 8f (0.014 g, 76%) [Found
(M+Na)+: 1422.3425, C91H178NaO8 requires: 1422.3414]; [α] 21
D
+10 (c 0.10, CHCl3); δH (400 MHz, CDCl3): 4.89 (1H, s), 4.51
(1H, dd, J 12, 3.9 Hz), 4.33 (1H, dd, J 12.0, 4.1 Hz), 4.21 – 4.15
(1H, m), 4.07 (1H, br. s), 4.01 – 3.95 (1H, m), 3.75 (1H, d, J 5.3
Hz), 3.73 – 3.66 (1H, m), 3.41 (3H, s), 3.35 (3H, s), 3.00 – 2.93
(1H, m), 2.85 – 2.70 (2H, m), 2.50 – 2.39 (1H, m), 1.72 – 1.61
(2H, m), 1.60 – 1.03 (145H, m), 0.89 (6H, t, J 6.8 Hz), 0.85 (3H,
d, J 6.8 Hz), 0.70 – 0.61 (2H, m), 0.60 – 0.53 (1H, dt, J 8, 4 Hz),
-0.33 (1H, br. q, J 5.2 Hz); δC (101 MHz, CDCl3): 175.0, 108.7,
85.5, 83.8, 80.4, 78.4, 72.8, 63.2, 57.7, 55.0, 52.2, 35.3, 35.2,
32.3, 31.9, 30.5, 30.2, 29.9, 29.89, 29.7, 29.6, 29.6, 29.5, 29.4,
29.3, 28.7, 27.6, 27.4, 26.1, 22.4, 22.7, 15.77, 14.8, 14.1, 10.9;
νmax: br. 3436, 2921, 2852, 1732, 1493, 1455, 759 cm-1.
4.89 (1H, s), 4.50 (1H, dd, J 12, 3.8 Hz), 4.32 (1H, dd, J 12, 4.4
Hz), 4.20 – 4.16 (1H, m), 4.07 (1H, d, J 5.6 Hz), 4.00 – 3.96 (1H,
m), 3.74 – 3.64 (1H, m), 3.41 (3H, s), 3.35 (3H, s), 3.00 – 2.93
(1H, m), 2.79 – 2.7 (2H, m), 2.48 – 2.39 (1H, m), 2.39 – 2.32
(1H, m), 1.6 – 1.05 (143H, m), 0.89 (6H, t, J 6.8 Hz), 0.86 (3H,
d, J 6.9 Hz), 0.69 – 0.63 (2H, m), 0.60 – 0.53 (1H, dt J ), -0.32
(1H, br. q J 4.5 Hz); δC (101 MHz, CDCl3): 174.9, 108.7, 85.4,
83.8, 80.4, 78.4, 72.8, 63.2, 57.7, 55.0, 52.2, 35.3, 35.2, 32.3,
31.9, 30.5, 30.2, 30.0, 29.9, 29.7, 29.68, 29.6, 29.5, 29.4, 29.3,
28.7, 27.5, 27.4, 26.1, 25.4, 22.7, 15.7, 14.99, 14.1, 10.9; νmax: br.
3436, 2918, 2850, 1732, 1467, 1099, 720 cm-1.
3.11: Methyl 5-O-(2-[(1R)-1-hydroxy-16-[(1R,2S)-2-[20-
methyl-19-oxooctatriacontyl]cyclopropyl] hexadecyl] tetra-
cosanoate) α-D-Araf 8g
(a) Cesium hydrogen carbonate (0.086 g, 0.443 mmol) was added
to a stirred solution of 5 (0.0475 g, 0.0953 mmol) and (R)-2-{(R)-
1-hydroxy-16-[(1R,2S)-2-((S)-20-methyl-19-oxo-octatriacontyl)-
cyclopropyl]hexadecyl}tetracosanoic acid (0.077 g, 0.063
mmol)41 in dry DMF:THF (1:5, 2 mL) at room temperature and
stirred at 70 ºC for two days. Work up as before gave methyl 2,3-
di-O-benzyl-5-O-(2-[(1R)-1-hydroxy-16-[(1R,2S)-2-[20-methyl-
19-oxooctatriacontyl]cyclopropyl]hexadecyl]tetracosanoate) α-
3.10: Methyl 5-O-(2-[(R)-1-hydroxy-18-[(1R, 2S)-2-[(17S, 18
S)-17-methoxy-18-methylhexatriacontyl]cyclopropyl]octa-
decyl] hexacosanoate) α-D-Araf 8f
D
-Araf as a thick colorless oil 7g (0.077 g, 80%) [Found
(M+Na)+: 1558.3708, C102H182NaO8 requires: 1558.3727], [α] 21
D
(a) Cesium hydrogen carbonate (0.083 g, 0.428 mmol) was
added with stirring to 5 (0.0457 g, 0.091 mmol) and (R)-2-{(R)-
1-hydroxy-18-[(1R,2S)-2-((17S,18S)-17-methoxy-18-
methylhexa- triacontyl)cyclopropyl]octadecyl}hexacosanoic acid
(0.076 g, 0.060 mmol)40 in dry DMF:THF (1:5, 2 mL) at room
temperature and the mixture was stirred at 70 ºC for two days
then worked up as before to give methyl 2,3-di-O-benzyl-5-O-(2-
[(R)-1-hydroxy-18-[(1R,2S)-2-[(17S,18S)-17-methoxy-18-
methylhexatriacontyl]- cyclopropyl]octadecyl]hexacosanoate)α-
+21 (c 0.10, CHCl3); δH (400 MHz, CDCl3): 7.39 – 7.28 (10H,
m), 4.92 (1H, s), 4.58 (1H, d, J 12.0 Hz), 4.56 (1H, d, J 12 Hz),
4.51 (1H, d, J 12 Hz), 4.48 (1H, d, J 12 Hz), 4.32 – 4.28 (2H, m),
4.25 – 4.19 (1H, m), 3.99 (1H, br. d, J 2.2 Hz), 3.84 (1H, dd, J
6.4, 2.8 Hz), 3.67 – 3.59 (1H, m), 3.38 (3H, s), 2.57 – 2.47 (2H,
m), 2.47 – 2.38 (3H, m), 1.75 – 1.09 (140H, m), 1.06 (3H, d, J
6.9 Hz), 0.89 (6H, t, J 6.7 Hz), 0.70 – 0.62 (2H, m), 0.61 – 0.53
(1H, dt, J 3.9, 7.9 Hz), -0.32 (1H, br. q, J 5.2 Hz); δC (101 MHz,
CDCl3): 215.2, 175.0, 137.5, 137.3, 128.5, 128.4, 127.9, 127.9,
127.8, 107.2, 87.8, 83.7, 79.4, 72.4, 72.1, 63.4, 54.9, 51.5, 46.3,
41.1, 33.0, 31.9, 30.2, 29.7, 29.6, 29.6, 29.5, 29.4, 29.3, 28.7,
27.4, 27.3, 25.7, 23.7, 22.7, 16.3, 15.7, 14.1, 10.9; νmax: br. 3524,
3030, 3063, 2922, 2851, 1732, 1715, 1465, 1107, 733 cm-1.
D
-Araf 7f (0.077 g, 80%) as a thick colorless oil [Found
(M+Na)+: 1602.4304, C105H190NaO8 requires: 1602.4353], [α] 22
D
+18 (c 0.10, CHCl3), which showed δH (400 MHz, CDCl3): 7.39
– 7.28 (10H, m), 4.92 (1H, s), 4.58 (1H, d, J 12 Hz), 4.56 (1H, d,
J 12 Hz), 4.51 (1H, d, J 12 Hz), 4.48 (1H, d, J 12 Hz), 4.31 –
4.28 (2H, m), 4.26 – 4.19 (1H, m), 4.0 (1H, dd, J 5.6, 2.5 Hz),
3.84 (1H, dd, J 6.3, 2.5 Hz), 3.68 – 3.58 (1H, m), 3.37 (3H, s),
(b) Palladium hydroxide on activated charcoal (20% Pd(OH)2-C,
0.0015 g, 0.15 fold by weight) was added to a stirred solution of
7g (0.010 g, 0.006 mmol) in dry CH2Cl2 : MeOH (1:1, 2 mL) at
room temperature under hydrogen atmosphere. The reaction

9
mixture was stirred overnight then worked up as before to afford
3.13: Methyl 5-O-(2-{(1R)-1-hydroxy-17-[(1S, 2R)-2-[(2S)-
22-methyl-21-oxotetracontan-2-yl]cyclopropyl]
heptadecyl}hexa- cosanoate) α-D-Araf 8h
ACCEPTED MANUSCRIPT
the title compound as a thick white oil 8g (0.0069 g, 63%)
[Found (M+Na)+: 1378.2788, C88H170NaO8 requires: 1378.2803],
[α] 2D1 +10.5 (c 0.10, CHCl3 + few drops of CD3OD); δH (400
(b) Cesium hydrogen carbonate (0.045 g, 0.232 mmol) was added
with stirring to 5 (0.025 g, 0.050mmol) and (R)-2-{(R)-17-[(1S,2R)-
2-((1S)-1,21-dimethyl-20-oxo-nonatriacontyl)cyclopropyl]-1-
hydroxy heptadecyl}hexacosanoic acid methyl ester (0.043 g, 0.033
mmol)43 in dry DMF:THF (1:5, 2 mL) at room temperature and
stirred at 70 ºC for two days, then worked up as before to give methyl
2,3-di-O-benzyl-5-O-(2-{(1R)-1-hydroxy-17-[(1S,2R)-2-[(2S)-22-
methyl-21-oxotetra-contan-2-
MHz, CDCl3): 4.89 (1H, s), 4.51 (1H, dd, J 12, 4.0 Hz), 4.33
(1H, dd, J 12.0, 4.1 Hz), 4.21 – 4.15 (1H, m), 4.07 (1H, br. s),
4.02 – 3.95 (1H, m), 3.74 – 3.66 (1H, m), 3.41 (3H, s), 2.90 –
2.71 (2H, m), 2.56 – 2.48 (1H, m), 2.48 – 2.37 (4H, m, including
the OH), 2.23 – 1.10 (140H, m), 1.05 (3H, d, J 6.9 Hz), 0.89 (6H,
t, J 6.8 Hz), 0.70 – 0.61 (2H, m), 0.59 – 0.53 (1H, dt, J 7.9, 3.9
Hz), -0.33 (1H, br. q, J 5.2 Hz); δC (101 MHz, CDCl3 + few drops
of CD3OD): 215.0, 174.9, 108.7, 83.8, 80.4, 78.4, 72.8, 63.2,
54.9, 52.1, 46.3, 41.1, 35.2, 33.0, 31.9, 30.2, 29.7, 29.6, 29.5,
29.45, 29.4, 29.35, 29.3, 28.7, 27.4, 27.3, 25.4 23.7, 22.7, 16.36,
15.7, 14.1, 10.9 ; νmax: br. 3436, 2918, 2850, 1731, 1708, 1467,
1170 cm-1.
yl]cyclopropyl]heptadecyl}hexacosanoate) α-D- Araf 7h as a thick
colorless oil (0.050 g, 92%) [Found (M+Na)+:1628.4539,
C107H192NaO8 requires: 1628.4509], [α] 21 +16.8 (c 0.1, CHCl3); δH
D
(400 MHz, CDCl3): 7.40 – 7.27 (10H, m), 4.92 (1H, s), 4.58 (1H, d,
J 12.0 Hz), 4.56 (1H, d, J 12 Hz), 4.53 (1H, d, J 12 Hz), 4.48 (1H, d,
J 12 Hz), 4.33 – 4.28 (2H, m), 4.22 (1H, dt, J 10.7, 4.6 Hz), 3.99 (1
H, br d, J 1.9 Hz), 3.84 (1H, dd, J 6.4, 2.6 Hz), 3.68 – 3.58 (1H, dt, J
7.6, 5.2 Hz), 3.38 (3H, s), 2.58 – 2.47 (2H, m), 2.47 – 2.38 (3H,
m), 1.72 – 1.13 (146H, m), 1.06 (3H, d, J 6.9 Hz), 0.89 (9H,
including a triplet J 7.5 Hz), 0.72 – 0.61 (1H, m), 0.50 – 0.41
(1H, m), 0.24 – 0.08 (3H, m); δC (101 MHz, CDCl3): 215.2,
175.0, 137.5, 137.3, 128.5, 128.4, 128.0, 127.9, 127.8, 107.2,
87.8, 83.7, 79.4, 72.4, 72.1, 63.5, 54.9, 51.5, 46.3, 41.1, 38.1,
37.4, 35.5, 34.5, 33.0, 31.9, 29.7, 29.6, 29.5, 29.4, 29.3, 27.4,
27.35, 27.3, 26.1, 25.7, 23.7, 22.7, 19.7, 18.6, 16.3, 14.1, 10.5 ;
3.12: Methyl 5-O-(2-[(1R)-1-hydroxy-16-[(1R,2S)-2-(20-
methyl-19-oxo- octatriacontyl)cyclopropyl]hexadecyl]-
hexacosa-noate)-α-D-Araf 8g’
(a) Cesium hydrogencarbonate (0.062g, 0.319 mmol) was added
to a stirred solution of 5 (0.048 g, 0.096 mmol) and (R)-2-{(R)-
1-hydroxy-16-[(1R,2S)-2-((S)-20-methyl-19-oxo-octatriacontyl)-
cyclopropyl]hexadecyl}hexacosanoic acid
(0.080 g, 0.064
mmol)42 in dry DMF : THF (1:5, 4 mL) at room temperature and
the mixture was stirred at 70 ºC for 2 days, then worked up and
purified as before to afford methyl 2,3-di-O-benzyl-5-O-(2-[(1R)-
1-hydroxy-16-[(1R,2S)-2-(20-methyl-19-oxooctatriacontyl)-
ν
max: br. 3524, 3064, 3031, 2923, 2853, 1737, 1715, 1465, 1107,
734 cm-1.
(b) Palladium hydroxide on activated charcoal (20% Pd(OH)2-C
, 0.0015 g, 0.15 fold by weight) was added to a stirred solution of
7h (0.010 g, 0.006 mmol ) in dry CH2Cl2 : MeOH (1:1, 2 mL) at
room temperature under hydrogen atmosphere. The mixture was
stirred overnight then worked up as before to afford the title
compound as a thick colorless oil 8h (0.010 g, 80%) [Found
cyclopropyl]hexadecyl]hexacosanoate)-α-D-Araf as a colourless
thick oil 7g’ (72 mg, 71%) [Found (M+NH4)+: 1581.4467,
C104H190NO8 requires: 1581.4486], [α]23 + 25 (c 0.10, CHCl3)
D
which showed δH (400 MHz, CDCl3): 7.42 – 7.27 (10H, m), 4.92
(1H, s), 4.58 (1H, d, J 12.0 Hz), 4.56 (1H, d, J 11.8 Hz), 4.50
(1H, d, J 12.0 Hz), 4.47 (1H, d, J 11.8 Hz), 4.34 – 4.26 (2H, m),
4.22 (1H, dt, J 4.6, 9.1 Hz), 3.99 (1H, br. d, J 2.1 Hz), 3.84 (1H,
dd, J 2.6, 6.4 Hz), 3.68 – 3.58 (1H, m), 3.38 (3H, s), 2.56 – 2.47
(2H, m, including OH), 2.47 – 2.37 (3H, m), 1.76 – 1.09 (144H,
m), 1.05 (3H, d, J 6.9 Hz), 0.89 (6H, t, J 6.8 Hz), 0.70 – 0.61
(2H, m), 0.56 (1H, dt, J 4.0, 8.0 Hz), -0.33 (1H, br. q, J 5.2 Hz);
δC (101 MHz, CDCl3): 215.2, 175.0, 137.4, 137.2, 128.5, 128.4,
128.0, 127.9, 127.8, 107.2, 87.8, 83.7, 79.4, 72.4, 72.1, 72.0,
63.4, 54.9, 51.5, 46.3, 41.1, 35.5, 33.0, 31.9, 30.2, 29.7, 29.67,
29.6, 29.57, 29.5, 29.46, 29.4, 29.3, 28.7, 27.4, 27.3, 25.8, 23.7,
22.7, 16.4, 15.8, 14.1, 10.9; νmax: 3457, 3064, 3032, 2921, 2851,
1717, 1678, 1466, 1101, 755, 697 cm-1.
(M+Na)+: 1448.3585, C93H180NaO8 requires: 1448.3570], [α] 21
+
D
13 (c 0.10, CHCl3); δH (400 MHz, CDCl3): 4.89 (1H, s), 4.49
(1H, dd, J 12, 4 Hz), 4.33 (1H, dd, J 12, 4.2 Hz), 4.21 – 4.16 (1H,
m), 4.09 (1H, d, J 6.8 Hz), 4.01 – 3.95 (1H, m), 3.75 (1H, d, J 5.4
Hz), 3.73 – 3.65 (1H, m), 3.41 (3H, s), 2.61 – 2.47 (1H, m), 2.46
– 2.28 (5H, m), 1.72 – 1.15 (146H, m), 1.06 (3H, d, J 6.9 Hz),
0.89 (9H, including a triplet at 0.89 with J 7.4 Hz), 0.76 – 0.61
(1H, m), 0.51 – 0.39 (1H, m), 0.26 – 0.05 (3H, m); δC (101 MHz,
CDCl3): 215.3, 174.9, 108.7, 83.8, 80.4, 78.8, 78.4, 72.8, 54.9,
52.2, 46.3, 44.5, 41.1, 38.1, 37.4, 35.2, 34.5, 33.0, 31.9, 30.0,
29.7, 29.6, 29.5, 29.3, 27.3, 26.1, 25.4, 23.71, 22.7, 19.7, 18.6,
16.3, 14.1, 10.5; νmax: br. 3467, 2917, 2849, 1731, 1713, 1467,
1050,720 cm-1.
(b) Palladium hydroxide on activated charcoal (20% Pd(OH)2-C,
0.01g, 0.15 fold by weight) was added to a stirred solution of
compound 7g’ (0.066 g, 0.042 mmol) in dry CH2Cl2 : MeOH
(1:1, 2 mL) at room temperature under hydrogen atmosphere.
The mixture was worked up and purified as before to afford the
title compound 8g’ as a colourless thick oil (39 mg, 66%) [Found
3.14: Methyl 5-O-(2-{(R)-1-hydroxy-12-[(1S, 2R) -2-[14-[(1S,
2R)-2-eicosylcyclopropyl]tetradecyl]cyclopropyl]dodecyl}-
hexacosanoate) α-D-Araf 8i
(M+NH4)+: 1401.3546, C90H178NO8, requires: 1401.3547], [α]23
D
(a) Cesium hydrogen carbonate (0.089g, 0.46 mmol) was added
to a stirred solution of 5 (0.049 g, 0.098 mmol) and (R)-2-((R)-1-
hydroxy-12-{(1S,2R)-2-[14-((1S,2R)-2-eicosylcyclopropyl)tetradec-
yl]cyclopropyl}dodecyl)hexacosanoic acid (0.075 g, 0.065 mmol)37
in dry DMF:THF (1:5, 2 mL) at room temperature and then
stirred at 70º C for two days. Work up as before gave methyl 2,3-
di-O-benzyl-5-O-(2-{(R)-1-hydroxy-12-[(1S,2R)-2-[14-[(1S,2R)-
2-eicosylcyclopropyl]tetradecyl]cyclopropyl]dodecyl}hexacosan-
+11 (c 0.10, CHCl3) which showed δH (400 MHz, CDCl3 + few
drops CD3OD): 4.89 (1H, s), 4.51 (1H, dd, J 3.9, 11.9 Hz), 4.32
(1H, dd, J 4.0, 11.9 Hz), 4.21 – 4.15 (1H, m), 4.07 (1H, br. s),
3.98 (1H, br. d, J 1.8 Hz), 3.74 – 3.66 (1H, m), 3.41 (3H, s), 2.87
– 2.67 (2H, m), 2.55 – 2.38 (4H, m), 1.71 – 1.12 (145H, m), 1.05
(3H, d, J 6.9 Hz), 0.89 (6H, t, J 6.8 Hz), 0.69 – 0.61 (2H, m),
0.56 (1H, dt, J 4.0, 8.0 Hz), -0.33 (1H, br. q, J 5.2 Hz); δC (101
MHz, CDCl3 + few drops CD3OD): 215.4, 175.0, 108.7, 83.6
,
oate) α-D-Araf 7i as a thick colorless oil (0.084 g, 87%) [Found
80.6, 78.4, 72.8, 63.2, 55.0, 52.3, 50.9, 46.3, 41.1, 35.2, 33.0, 31.9,
30.2, 29.7, 29.65, 29.6, 29.5, 29.45, 29.41, 29.40, 29.3, 28.7, 27.4,
27.3, 25.4, 23.7, 22.7, 16.4, 15.8, 14.1, 10.9; νmax: 3405, 2920, 2851,
1713, 1712, 1466, 1108, 759 cm-1.
(M+Na)+: 1486.3173, C98H174NaO7 requires: 1486.3152], [α] 21
D
+31 (c 0.10, CHCl3); δH (400 MHz, CDCl3): 7.39 – 7.28 (10H,
m), 4.92 (1H, s), 4.58 (1H, d, J 12 Hz), 4.56 (1H, d, J 12 Hz),

10
Tetrahedron
ACCEPTED MANUSCRIPT
4.51 (1H, d, J 12 Hz), 4.48 (1H, d, J 12 Hz), 4.32 – 4.28 (2H,
MM wishes to thank the Ministry of Higher Education &
m), 4.25 – 4.19 (1H, m), 3.99 (1H, br d, J 2.3 Hz), 3.84 (1H, dd,
J 6.4, 2.6 Hz), 3.68 – 3.59 (1H, m), 3.38 (3H, s), 2.52 (1H, d, J
8.3 Hz), 2.44 (1H, dt, J 9.3, 5.5 Hz), 1.62 – 1.03 (134H, m), 0.89
(6H, t, J 6.7 Hz), 0.72 – 0.62 (4H, m), 0.61 – 0.52 (2H, dt J 8, 4
Hz), -0.32 (2H, br. q J 5.1 Hz); δC (101 MHz, CDCl3): 175.0,
137.4, 137.2, 128.5, 128.4, 128.0, 127.9, 127.89, 127.8, 107.2,
87.8, 83.7, 79.4, 72.4, 72.2, 72.1, 63.5, 54.9, 51.5, 35.5, 31.9,
30.55, 30.5, 30.45, 30.44, 30.4, 30.35, 30.3, 30.28, 30.2, 30.18,
30.1, 30.0, 29.9, 29.7, 29.6, 29.5, 29.4, 29.3, 29.29, 29.2, 29.1,
29.03, 29.0, 28.7, 28.6, 27.4, 25.7, 22.68, 22.6, 15.7, 14.1, 10.9 ;
Scientific Research - Kurdistan Region, Iraq for the award of a
grant. We thank the EPSRC UK National Mass Spectrometry
Facility at Swansea University for carrying out accurate mass
measurements. We thank Dr A Ramsay and the Special
Programme for Research and Training in Tropical Diseases at the
World Health Organization for access to materials from the TDR
TB Specimen Bank
References and notes
ν
max: 3479, 3065, 2989, 2919, 2849, 1733,1607, 1494, 718 cm-1.
1.
2.
3.
Brennan, P. J., Nikaido, H. The Envelope of Mycobacteria, Ann.
Rev. Biochem., 1995, 64, 29 – 63.
Mishra, A. K., Driessen, N. N., Appelmelk, B. J., Besra, G. S.
FEMS Microbiol., 2011, 35, 1126 – 1157.
Daffe, M., McNeil, M., Brennan, P. J. Carb. Res., 1993, 249, 383
– 398.
Brennan, P. J. Tuberculosis, 2003, 83, 91 – 97.
(b) Palladium hydroxide on activated charcoal (20% Pd(OH)2-C ,
0.0087 g , 0.15 fold by weight) was added to a stirred solution of
7i (0.058g, 0.039 mmol) in dry CH2Cl2 : MeOH (1:1, 2 mL) at
room temperature under hydrogen atmosphere. The mixture was
stirred overnight then worked up as before to afford the title
compound as a thick colourless oil 8i (0.040 g, 80%) [Found
(M+Na)+: 1306.2201, C84H162NaO7 requires: 1306.2213], [α] 2D3
4.
5.
Umesiri, F. E., Sanki, A. K., Boucau, J., Ronning, D. R., Sucheck,
S. J. Med. Res. Rev., 2010, 30, 290 – 326.
Miyauchi, M., Murata, M., Shibuya, K., Koga-Yamakawa, E.,
Uenishi, Y., Kusunose, N., Sunagawa, M., Yano, I., Kashiwazaki,
Y. Drug Disc. Ther., 2011, 5, 130 – 135.
6.
+ 12 (c 0.10, CHCl3); δH (400 MHz, CDCl3 + few drops of
CD3OD): 4.89 (1H, s), 4.47 (1H, dd, J 11.8, 4.2 Hz), 4.37 (1H,
dd, J 11.9, 4.1 Hz), 4.20 – 4.14 (1H, m), 4.07 (1H, br. s), 3.98
(1H, br. s), 3.79 – 3.67 (1H, m), 3.40 (3H, s), 3.06 (2H, br. s),
2.45 (1H, dt, J 10.1, 5.0 Hz), 1.90 – 1.83 (1H, m), 1.63 – 1.07
(134H, m), 0.89 (6H, t, J 6.8 Hz), 0.71 – 0.61 (4H, m), 0.60 –
0.49 (2H, dt, J 8, 4 Hz), -0.33 (2H, br. q J 5.2 Hz); δC (101 MHz,
CDCl3): 175.0, 108.7, 83.4, 80.7, 78.4, 72.8, 63.2, 54.9, 52.4,
7.
8.
9.
Janin, Y. L. Bioorg. Med. Chem., 2007, 15, 2479 – 2513.
Schroeder, E. K., de Souza, O. N., Santos, D. S., Blanchard, J. S.,
Basso, L. A. Curr. Pharm. Biotech., 2002, 3, 197 – 225.
Anderson, R. J. Chem. Rev., 1941, 29, 225 – 243.
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