Synthesis of D-1,2-dideoxy-1,2-difluoro-myo-inositol 3,4,5,6-tetrakisphosphate and its enantiomer as analogues of myo-inositol 3,4,5,6-tetrakisphosphate

Article abstract of DOI:10.1016/S0008-6215(98)00146-3

DL-3,4,5,6-Tetra-O-benzyl-1-deoxy-1-fluoro-scyllo-inositol was resolved using (-)-(1S, 4R)-camphanyl chloride. The diastereoisomers formed were separated and the structure of D-3,4,5,6-tetra-O-benzyl-2-(1S,4R)-camphanyl-1-deoxy-1-fluoro-scyllo-inositol wa

Full text of DOI:10.1016/S0008-6215(98)00146-3

Carbohydrate Research 309 (1998) 337±343
Synthesis of d-1,2-dideoxy-1,2-di¯uoro-myo-
inositol 3,4,5,6-tetrakisphosphate and its
enantiomer as analogues of myo-inositol
3,4,5,6-tetrakisphosphate
Kevin R.H. Solomonsa, Sally Freemana,*, Carl H. Schwalbeb,
Stephen B. Shearsc, Deborah J. Nelsond, Weiwen Xied, Karol S. Bruzike,
Marcia A. Kaetzelf
a School of Pharmacy and Pharmaceutical Sciences, University of Manchester, Oxford Road,
Manchester M13 9PL, UK
b Department of Pharmaceutical and Biological Sciences, Aston University, Birmingham B4 7ET,
UK
c Inositide Signaling Section, Laboratory of Signal Transduction, National Institute of Environmental
Health Sciences, National Institutes of Health, Research Triangle Park, NC 27709, USA
d Department of Neurology, University of Chicago, 5841 S Maryland Avenue, Chicago,
IL 60637, USA
e DepartmentofMedicinalChemistryandPharmacognosy,CollegeofPharmacy,UniversityofIllinoisat
Chicago, Chicago IL 60612-7231, USA
f Department of Molecular and Cellular Physiology, College of Medicine, University of Cincinnati, OH
45267-0576, USA
Received 23 March 1998; accepted 9 June 1998
Abstract
dl-3,4,5,6-Tetra-O-benzyl-1-deoxy-1-¯uoro-scyllo-inositol was resolved using ( )-(1S, 4R)-
camphanyl chloride. The diastereoisomers formed were separated and the structure of d-3,4,5,6-
tetra-O-benzyl-2-(1S,4R)-camphanyl-1-deoxy-1-¯uoro-scyllo-inositol was solved by X-ray
crystallography to an R-factor of 4.2%. A series of manipulations led to the preparation of d-1,2-
dideoxy-1,2-di¯uoro-myo-inositol 3,4,5,6-tetrakisphosphate and its enantiomer. The d-1,2-
di¯uoro enantiomer stereospeci®cally inhibited CaMK II-activated Cl current, but with low
potency; however, ecacy of this compound was greatly enhanced by myo-inositol 3,4,5,6-tetra-
kisphosphate itself. # 1998 Elsevier Science Ltd. All rights reserved
Keywords: Inositol polyphosphates; Chloride current; Myo-inositol 3,4,5,6-tetrakisphosphate; Crystal structure
* Corresponding author. Fax: 0044 (0)161 275 2396; e-mail: sfreeman@fsl.pa.man.ac.uk
0008-6215/98/$Ðsee front matter # 1998 Elsevier Science Ltd. All rights reserved
PII S0008-6215(98)00146-3

338
K.R.H. Solomons et al./Carbohydrate Research 309 (1998) 337±343
1. Introduction
Since the ®nding that myo-inositol 1,4,5-trisphos-
phate is the second messenger linking receptor
activation to the mobilisation of calcium from
intracellular stores [1], there has been widespread
investigation into the metabolism and e€ects of
inositol phosphates as mediators of cell signalling
transduction pathways [2,3]. d-myo-Inositol
3,4,5,6-tetrakisphosphate (Ins[3,4,5,6]P4, 1a) has
been shown to be involved in the long-term
uncoupling of chloride secretion from intracellular
calcium levels [4]. More recently it has been
demonstrated that 1a inhibits calcium-dependent
chloride conductance, which is important in the
cellular control of salt and ¯uid secretion, and may
have implications in the treatment of cystic ®brosis
[5]. Tetrakisphosphate 1a has also been shown to
be a potent inhibitor of myo-inositol 1,3,4-trisphos-
phate 5/6-kinase activity [6]. In addition, levels of
its enantiomer, d-myo-inositol 1,4,5,6-tetrakispho-
sphate (Ins[1,4,5,6]P4, 1b), may increase after cell
transformation with the src oncogene [7], and also
after treatment with phosphoinositidase (PIC)-
linked agonists [8].
Scheme 2. Synthesis of d-1,2-dideoxy-1,2-di¯uoro-myo-inositol
3,4,5,6-tetrakisphosphate (2a). Reagents: (a) aq NaOH,
MeOH; (b) DAST, CH2Cl2, Et3N; (c) H2, 10% Pd-C, THF±
H2O (1:1); (d) (BnO)2PNPri2, 1H-tetrazole, THF, then MCPBA.
whereas hydroxy can both accept and donate [9].
We have previously reported the synthesis of racemic
dl-1,2-dideoxy-1,2-di¯uoro-myo-inositol 3,4,5,6-
tetrakisphosphate (2) [10], and here the enantio-
mers 2a and 2b have been synthesised. Enantiomer
2a represents the ®rst biologically-active analogue
of 1a. Schultz and coworkers have recently repor-
ted the synthesis of the corresponding 1,2-dichloro
and 1,2-dimethoxy analogues [11,12].
2. Results and discussion
The synthesis of 2a is shown in Schemes 1 and 2.
The enantiomer 2b was prepared analogously from
4b.
dl-3,4,5,6-Tetra-O-benzyl-1-deoxy-1-¯uoro-
Fluorine is a sterically conservative replacement
for a hydroxy group: the signi®cant di€erence is
that ¯uorine can only accept hydrogen bonds,
scyllo-inositol 3 was prepared in three steps from
dl-3,4,5,6-tetra-O-benzyl-myo-inositol [10]. Cam-
phanyl esters have been useful in determining the
absolute con®guration of inositol derivatives [13±
15] and here the resolution of 3 was achieved by
treatment with ( )-(1S, 4R)-camphanyl chloride
(Scheme 1). The diastereoisomeric (1S,4R)-cam-
phanyl esters 4a and 4b were readily separated by
¯ash chromatography and were fully characterised,
1
with data including H and 13C NMR spectra, ele-
mental analysis and speci®c optical rotation mea-
surements. The second eluted diastereoisomer
crystallised from diethyl ether±hexane to give
X-ray quality crystals. The structure was re®ned to
R=4.2%.1 The atomic coordinates are given in
Table 1 and Fig. 1 gives the molecular geometry.
Correct stereochemistry was con®rmed by the
known camphanyl geometry and the structure was
determined as diastereoisomer 4a. The ®ve inositol
oxygens and the ¯uoro substituent are all equator-
ial, as required for the scyllo isomer. The inositol
ring adopts a distorted, locally ¯attened chair
1 Tables of atomic coordinates, bond lengths, and bond
angles have been deposited with the Cambridge Crystal-
lographic Data Centre. These tables may be obtained, on
request, from the Director, Cambridge Crystallographic Data
Centre, 12 union Road, Cambridge, UK, CB2 1EZ.
Scheme 1. Resolution of dl-3,4,5,6-tetra-O-benzyl-1-deoxy-1-
¯uoro-scyllo-inositol. Reagents: (a) ( )-(1S,4R)-camphanyl
chloride, DMAP, Et3N, CH2Cl2.

K.R.H. Solomons et al./Carbohydrate Research 309 (1998) 337±343
339
conformation: the torsion angles around the ring
vary in magnitude from a maximum for C1±C2±
C3±C4 [55.9(4)o] to a minimum for C4±C5±C6±C1
[ 49.3(4)o], compared with the consensus value of
56ꢀ in cyclohexane [17]. The best-preserved symmetry
elements of a cyclohexane chair are the mirror
plane through C2 with asymmetry parameter [18]
ÁCS=1.7ꢀ and the two-fold axis through the mid-
point of C2 and C3 with ÁC2=1.5ꢀ. The C±C
inositol ring bond lengths (range 1.499±1.529 A)
are comparable with those of myo-inositol [19].
The C±C±C inositol ring bond angles (range 109.9±
113.0ꢀ) are close to those of a perfect chair [17].
Alkaline hydrolysis of esters 4a and 4b yielded
the enantiomeric alcohols 3a and 3b in quantitative
yield, which were converted to the dideoxy-
di¯uoro-myo-inositol tetrakisphosphates, 2a and
2b, by methods previously described for the race-
mate of 2 (Scheme 2, shown for a) [10]. This
required ¯uorination of 3a and 3b with DAST to
give the di¯uoro compounds 5a and 5b, debenzy-
lation by hydrogenolysis to give the tetraols 6a and
6b, phosphorylation with dibenzyl N,N-diisopro-
pylphosphoramidite in the presence of 1H-tetra-
zole, followed by MCPBA to give the protected
tetrakisphosphates 7a and 7b, and subsequent
removal of the P-OBn groups by hydrogenolysis to
give the tetrakisphosphates 2a and 2b.
Table 1
Atomic coordinates (Â104) and equivalent isotropic displace-
ment parameters (A2Â103) for 4a. U(eq) is de®ned as one third
of the trace of the orthogonalized Uij tensor
x
y
z
U(eq)
C(2)
C(1)
C(6)
C(5)
C(4)
C(3)
F(8)
O(7)
O(12)
O(11)
O(10)
O(9)
7126(2)
6239(2)
5218(2)
4930(2)
5855(2)
6852(2)
8016(2)
6529(2)
4414(2)
4087(2)
5538(2)
7691(2)
6821(3)
6844(3)
7122(3)
7327(2)
7959(4)
8123(4)
8179(3)
6333(4)
7032(5)
8246(3)
9044(5)
9114(3)
8172(6)
8361(4)
9230(3)
10031(3)
10822(3)
10821(5)
10056(5)
9262(3)
5679(4)
5296(3)
5845(4)
5499(5)
4575(5)
4010(4)
4361(4)
3274(3)
2353(2)
2120(3)
1252(4)
599(3)
5608(4)
5898(3)
5188(3)
5449(4)
5212(4)
5949(4)
6326(3)
5401(2)
5694(3)
4554(3)
5649(2)
5541(2)
6305(4)
7484(3)
5634(4)
6693(3)
6154(5)
4719(5)
4844(4)
4692(6)
4093(6)
6803(4)
4048(7)
5712(8)
3515(6)
6584(5)
5957(4)
5261(5)
4679(6)
4818(7)
5519(8)
6090(5)
4670(5)
5246(4)
6217(6)
6738(7)
6285(8)
5313(8)
4799(7)
5077(4)
5636(4)
6966(4)
7476(5)
6659(5)
5305(5)
4794(4)
4724(5)
5399(5)
6564(5)
7152(7)
6563(11)
5391(12)
4796(8)
4786(2)
4122(2)
4318(2)
5228(2)
5897(2)
5690(2)
4575(2)
3304(1)
3717(1)
5390(2)
6710(1)
6284(1)
2721(2)
2844(2)
1925(2)
1313(2)
734(2)
59(1)
54(1)
55(1)
55(1)
56(1)
59(1)
88(1)
61(1)
65(1)
65(1)
64(1)
67(1)
64(1)
103(1)
60(1)
76(1)
79(1)
85(1)
76(1)
101(2)
117(2)
112(1)
134(2)
111(2)
135(3)
100(2)
65(1)
82(1)
108(2)
118(2)
113(2)
87(1)
84(1)
72(1)
102(2)
119(2)
122(2)
122(2)
100(1)
71(1)
58(1)
80(1)
102(2)
90(1)
94(1)
83(1)
85(1)
72(1)
94(1)
132(2)
142(3)
144(3)
112(2)
C(13)
O(14)
C(15)
O(16)
C(17)
C(18)
C(19)
C(20)
C(21)
O(22)
C(23)
C(24)
C(25)
C(26)
C(27)
C(28)
C(29)
C(30)
C(31)
C(32)
C(33)
C(34)
C(35)
C(36)
C(37)
C(38)
C(39)
C(40)
C(41)
C(42)
C(43)
C(44)
C(45)
C(46)
C(47)
C(48)
C(49)
C(50)
C(51)
C(52)
C(53)
An alternative preparation of 4a and 4b is out-
lined in Scheme 3. This requires the reaction of
dl-3,4,5,6-tetra-O-benzyl-myo-inositol (8) with ( )-
(1S,4R)-camphanyl chloride. Selectivity was
observed at the 1-/3-positions to give a mixture of
diastereoisomers 9a and 9b. Separation was
achieved by crystallisation, however it was more
968(2)
1962(2)
1444(3)
763(3)
150(2)
560(3)
2321(3)
2429(3)
6628(3)
7196(2)
6846(3)
7371(4)
8240(5)
8587(3)
8073(3)
7381(2)
8190(2)
8648(3)
9386(3)
9679(3)
9244(4)
8498(3)
5906(2)
5375(2)
5373(3)
4902(4)
4420(3)
4426(3)
4897(3)
3237(3)
2574(2)
2763(3)
2165(5)
1388(5)
1192(3)
1792(3)
804(4)
1666(3)
3829(3)
3085(3)
2555(3)
1851(4)
1627(5)
2135(5)
2850(4)
Fig. 1. ORTEP drawing [16] of 4a, showing the labelling
scheme for non-H atoms. Thermal ellipsoids are drawn at the
50% probability level.

340
K.R.H. Solomons et al./Carbohydrate Research 309 (1998) 337±343
contributes to the cooperative binding through
hydrogen-bond donation [20]. The inhibition was
stereospeci®c as the enantiomer 2b had no e€ect on
the CaMK II-activated Cl current [20]. In addi-
tion, here it is reported that 1a increases the
potency of inhibition of compound 2a: a 10 ꢀM
concentration of 2a had little e€ect on the CaMK
II-activated Cl current (Fig. 2, bar C), whereas
there was a synergistic decrease in the current when
6 ꢀM 2a was combined with 6 ꢀM 1a (Fig. 2, bar
D). This 1a-mediated increase in the potency of 2a
is consistent with inhibition of Cl current being
highly cooperative in vivo [5]. Thus, compound 2a
has contributed to de®ning the importance of
hydrogen-bonding in the cooperative inhibition of
1a, which should assist in the design of agonists
and antagonists for potential therapeutic applica-
tions. The major route of metabolism of 1a in vivo
is by phosphorylation by a 1-OH kinase [21].
Therefore compound 2a also has a potential appli-
cation as a metabolically stable analogue.
Scheme 3. Alternative syntheses of camphanyl esters 4a and
4b. Reagents: (a) ( )-(1S,4R)-camphanyl chloride, DMAP,
Et3N, CH2Cl2; (b) DAST, toluene.
convenient to ¯uorinate the diastereoisomeric
mixture with DAST to give4a and 4b in excellent yield,
which could be separated by ¯ash chromatography.
Most of the biological studies conducted with 2a
and 2b have been reported [20]. Brie¯y myo-inosi-
tol 1,4,5-trisphosphate causes the release of cyto-
solic calcium which binds to calmodulin and
activates calmodulin dependant protein kinase II
(CaMK II). Through phosphorylation this acti-
vates one type of chloride conductance (gClCaMK
)
(Fig. 2, bar A). This chloride current is down-
regulated by the tetrakisphosphate 1a (Fig. 2, bar
B) with an IC50 of 6 ꢀM [5], which shows non-
cooperative binding at low concentration and
cooperative binding at high concentration [20]. The
1,2-di¯uoro analogue 2a inhibited the chloride
current to provide the ®rst agonist of 1a, albeit
with lower anity (IC50 100 ꢀM) [20]. In contrast,
2a showed non-cooperative binding only, suggest-
ing that one or both of the hydroxy groups in 1a
3. Experimental
General.ÐInstrumentation, techniques and
reagent suppliers were as previously described [10]
with the following addition: optical rotations were
measured on an Optical Activity AA-100 polari-
ꢀ
meter using a 0.25 dm cell at 25 C. For com-
pounds 3a/b, 5a/b, 7a/b and 2a/b only brief
experimental information and 1H NMR data,
melting points and speci®c optical rotations are
given: the procedures and other spectral data (13C
and 19F NMR) were identical to those reported for
the racemic materials [10].
Crystal structure determination of (4a).Ð
C44H47FO8, M=722.8. Monoclinic, a=12.646(2),
b=9.939(1), c=15.611(2) A, ꢁ=94.107(9)ꢀ, V=
1957.0(4) A3 (by least squares analysis of setting
angles of 25 re¯ections, 23.0ꢁꢂꢁ27.0ꢀ, l=1.54178
3
A), space group P21, Z=2, Dc=1.227 Mg m .
Needle with dimensions: 0.4Â0.15Â0.05 mm,
1
ꢀ=0.71 mm .
Enraf-Nonius CAD4 di€ractometer, !-2ꢂ scan
technique, graphite-monochromated Cu radiation:
4262 re¯ections measured (2ꢁꢂꢁ67ꢀ, for 1ꢁhꢁ15,
0ꢁkꢁ11, 18ꢁlꢁ18), 3689 unique, giving 2895
with F>4ꢃ. The structure was solved by direct
methods [22] and re®ned by full-matrix least-
squares [23] with anisotropic displacement para-
meters for non-H atoms, and H atoms in calculated
Fig. 2. The e€ect of 2a upon the chloride current (ICl,CaMK
)
when added together with 1a. The mean (n=3) peak current
amplitudes at 110 mV are given after 12±15 min treatment with
CaMK II plus the following concentrations of 1a and 2a: bar
A-CaMK II activated (control); bar B-6 ꢀM 1a; bar C-10 ꢀM
2a; bar D-6 ꢀM 2a+6 ꢀM 1a.

K.R.H. Solomons et al./Carbohydrate Research 309 (1998) 337±343
341
positions. Final discrepancy indices are R=4.2%
for observed data and 6.4% for all data. Correct
stereochemistry is con®rmed by the known cam-
phanyl geometry and an absolute structure para-
meter [23] value of 0.2(2).
d-3,4,5,6-Tetra-O-benzyl-2-(10S,40R)-camphanyl-1-
deoxy-1-¯uoro-scyllo-inositol (4a) and d-1,4,5,6-
tetra-O-benzyl-2-(10S,40R)-camphanyl-3-deoxy-3-
¯uoro-scyllo-inositol (4b).ÐA solution of ( )-
(1S,4R)-camphanyl chloride (212 mg, 0.978 mmol)
in dichloromethane (1.2 mL) was added dropwise
to a stirred solution of dl-3,4,5,6-tetra-O-benzyl-1-
1.12 (3 H, s, CH3), 1.55±1.7 (1 H, m, Camph-CH2),
1.75±1.95 (2 H, m, Camph-CH2), 2.35±2.5 (1 H, m,
Camph-CH2), 3.5±3.8 (4 H, m, 3-, 4-,5- and 6-H),
2
4.54 (1 H, dt, JHF 51.8, J1/2ꢂJ1/6ꢂ9.6, 1-H), 4.7±
3
4.95 (8 H, m, 4ÂCH2Ph), 5.40 (1 H, br dt, JHF
11.9, J2/1ꢂJ2/3ꢂ9.9, 2-H), 7.2±7.35 (20 H, m, Ph);
13C NMR data (CDCl3): ꢅ 9.7 (CH3), 16.4
(2ÂCH3), 28.8 (CH2), 30.5 (CH2), 54.7 (quat.), 54.8
2
(quat.), 73.3 (d, JCF 17.1, C-2 or C-6), 75.4, 75.7,
75.95, 76.1 [4ÂCH2Ph], 78.9 (d, 3JCF 9.8, C-3 or C-
2
3
5), 80.5 (d, JCF 15.9, C-6 or C-2), 81.1 (d, JCF
11.0, C-5 or C-3), 82.1 (s, C-4), 91.0 (quat.), 92.7
1
deoxy-1-¯uoro-scyllo-inositol
3
[10] (265 mg,
(d, JCF 185.5, C-1), 127.3±128.4 (20Âarom CH),
0.489 mmol), DMAP (10 mg, 0.082 mmol) and
triethylamine (0.125 mL, 0.897 mmol) in anhydrous
137.75 (2Âarom C), 137.9 (arom C), 138.0 (arom
C), 166.8 (C=O), 178.3 (C=O); MS data (CI): 740
(M+NH4+, 40%), 108 (35), 91(100). Anal. Calcd
for C44H47FO8.0.5 H2O: C, 72.2; H, 6.6. Found C,
72.3; H, 6.4.
ꢀ
dichloromethane (6 mL) at 0 C under an atmo-
sphere of nitrogen. The mixture was allowed to
warm to room temp. and then stirred for 24 h. The
reaction mixture was washed with saturated aq.
NaHCO3 (2Â10 mL) and water (2Â10 mL), dried
(Na2SO4) and the solvent was evaporated o€ in
vacuo. Flash chromatography on silica eluting with
diethyl ether±dichloromethane (2:98) gave 4a (Rf
0.30) and 4b (Rf 0.41).
Alternatively, 4a and 4b were prepared directly
by heating a diastereoisomeric mixture of 3,4,5,6-
tetra-O-benzyl-1-(10S,40R)-camphanyl-myo-inositol
9a and 1,4,5,6-tetra-O-benzyl-3-(10S,40R)-campha-
nyl-myo-iꢀnositol 9b with DAST (16 eq) in toluene
at 70±80 C for 2 h. Aqueous work-up, ¯ash chro-
matography and recrystallisation as described
above gave 4a and 4b in 48 and 25% yields,
respectively.
Diastereoisomer 4a was recrystallised from
diethyl ether±hexane to give needles which were
suitable for X-ray crystallography (125 mg, 36%);
ꢀ
mp 112±113 C; [ꢄ]d +1.1‹0.6ꢀ (c 0.0143 g/mL,
d-3,4,5,6-Tetra-O-benzyl-1-deoxy-1-¯uoro-scyllo-
inositol (3a) and d-1,4,5,6-tetra-O-benzyl-3-deoxy-
3-¯uoro-scyllo-inositol (3b).ÐAqueous sodium
hydroxide (3 M, 2 mL) was added to a stirred
solution of each camphanyl ester, 4a or 4b (160 mg,
0.29 mmol) in warm methanol (15 mL). After 18 h
at room temp., methanol was removed by eva-
poration in vacuo and the residue was partitioned
between dichloromethane (2Â30 mL) and water
(30 mL). The combined organic layers were washed
with water (30 mL), dried (Na2SO4), and the sol-
vent evaporated to give the alcohols 3a or 3b in
quantitative yields. Recrystallisation from ethyl
acetate±hexane gave crystals of 3a; mp 121±122 ꢀC;
1
CHCl3); H NMR data (CDCl3): ꢅ 0.89 (3 H, s,
CH3), 1.05 (3 H, s, CH3), 1.10 (3 H, s, CH3), 1.6±
1.75 (1 H, m, Camph-CH2), 1.85±2.0 (2 H, m,
Camph-CH2), 2.3±2.4 (1 H, m, Camph-CH2), 3.5±
3.8 (4 H, m, 3-, 4-, 5- and 6-H), 4.51 (1 H, dt, 2JHF
51.5, J1/2ꢂJ1/6ꢂ9.3, 1-H), 4.65±4.95 (8 H, m,
3
4ÂCH2Ph), 5.42 (1 H, br dt, JHF 11.2, J2/1ꢂJ2/3
ꢂ9.9, 2-H), 7.2±7.35 (20 H, m, Ph); 13C NMR data
(CDCl3): ꢅ 9.6 (CH3), 16.3 (CH3), 16.7 (CH3),
28.95 (CH2), 30.7 (CH2), 54.2 (quat.), 54.7 (quat.),
2
73.0 (d, JCF 18.3, C-2 or C-6), 75.4, 75.5, 76.0,
3
76.1 [4ÂCH2Ph], 78.7 (d, JCF 11.0, C-3 or C-5),
2
3
80.4 (d, JCF 15.8, C-6 or C-2), 81.1 (d, JCF 12.2,
C-5 or C-3), 82.2 (s, C-4), 90.8 (quat.), 92.95 (d,
1JCF 186.7, C-1), 127.1±128.4 (20Âarom CH),
137.6 (arom C), 137.7 (arom C), 137.8 (arom C),
138.0 (arom C), 166.4 (C=O), 177.9 (C=O); MS
data (CI): 740 (M+NH4+, 14%), 216 (40), 108
(51), 91(100). Anal. Calcd for C44H47FO8: C, 73.1;
H, 6.55. Found: C, 72.9; H, 6.6.
[ꢄ]d
7.75‹0.3ꢀ (c 0.0258 g/mL, CHCl3); 1H
NMR (CDCl3): ꢅ 2.49 (1 H, s, OH), 3.40 (1 H, t, J4/3
ꢂJ4/5ꢂ9.0, 4-H), 3.5±3.8 (4 H, m, 2-, 3-, 5- and 6-
H), 4.42 (1 H, dt, JHF 52.4, J1/2ꢂJ1/6ꢂ9.0, 1-H),
4.75±4.95 (8 H, m, 4ÂCH2Ph), 7.25±7.35 (20 H, m,
Ph). 3b; mp 122±123 ꢀC; [ꢄ]d +6.1‹1.0 (c
0.0085 g/mL, CHCl3).
Diastereoisomer 4b was recrystallised from
ethanol (118 mg, 34%); mp 139±140 ꢀC; [ꢄ]d
3.6‹0.5ꢀ (c 0.0135 g/mL, CHCl3); 1H NMR data
(CDCl3): ꢅ 0.99 (3 H, s, CH3), 1.04 (3 H, s, CH3),
d-3,4,5,6-Tetra-O-benzyl-1,2-dideoxy-1,2-di¯uoro-
myo-inositol (5a) and d- 1,4,5,6-tetra-O-benzyl-2,3-
dideoxy-2,3-di¯uoro-myo-inositol (5b).ÐRecrys-
tallisation from ethanol gave needles of 5a; mp

342
K.R.H. Solomons et al./Carbohydrate Research 309 (1998) 337±343
87±88 ꢀC; [ꢄ]d 20.0‹0.7 (c 0.0108 g/mL, CHCl3);
( )-(1S,4R)-camphanyl chloride and dl-3,4,5,6-
tetra-O-benzyl-myo-inositol 8 (2.37 g, 4.39 mmol)
using a procedure similar to that described for 4a/
b. Crystallisation from ethanol gave a mixture of
9a and 9b (1.54 g, 49%); 1H NMR data (CDCl3): ꢅ
0.89 (3 H, s, CH3a), 0.97 (3 H, s, CH3b), 1.01 (3 H,
s, CH3b), 1.08 (3 H, s, CH3a), 1.10 (3 H, s, CH3a),
1.11 (3 H, s, CH3b), 1.55±1.75 (1 H, m, Camph-
CH2), 1.8±2.0 (2 H, m, Camph-CH2), 2.25±2.4 (1
H, m, Camph-CH2), 3.54 (1 H, t, JHHꢂJHHꢂ8.9,
inositol H), 3.57 (1 H, d, J3/4 9.2, 3-H), 3.95
(1 H, t, JHHꢂJHHꢂ9.6, inositol Ha), 3.97 (1 H,
t, JHHꢂJHHꢂ9.6, inositol Hb), 4.12 (1 H, t,
JHHꢂJHHꢂ9.6, inositol Ha), 4.14 (1 H, t,
JHHꢂJHHꢂ9.6, inositol Hb), 4.29 (1 H, t, J2/1ꢂJ2/3
ꢂ2.3, 2-Hb), 4.32 (1 H, t, J2/1ꢂJ2/3ꢂ2.3, 2-Ha),
4.65±4.95 (9 H, m, 4ÂCH2Ph and 1-H), 7.2-7.4 (20
H, m, Ph).
3
1H NMR (CDCl3): ꢅ 3.45 (1 H, br dd, J3/F-2 28.0,
J3/4 9.5, 3-H), 3.47 (1 H, t, J5/4ꢂJ5/6ꢂ9.4, 5-H),
4
3.97 (1 H, td, J4/3ꢂJ4/5ꢂ9.6, J4/F-2 1.3, 4-H), 4.04
3
4
(1 H, dtd, J6/F-1 9.9, J6/1ꢂJ6/5ꢂ9.6, J6/F-2 1.3, 6-
2
3
H), 4.42 (1 H, dddd, JHF 46.5, JHF 27.7, J1/6 9.7,
J1/2 1.8, 1-H), 4.7±4.95 (8 H, m, 4ÂCH2Ph), 5.12 (1
2
3
H, ddt, JHF 53.1, JHF 9.6, J2/1ꢂJ2/3ꢂ1.6, 2-H),
ꢀ
7.25±7.35 (20 H, m, Ph). 5b; mp 92±93 C; [ꢄ]d
+17.7‹0.4 (c 0.0162 g/mL, CHCl3).
d-1,2-Dideoxy-1,2-di¯uoro-myo-inositol 3,4,5,6-
tetrakis(dibenzyl phosphate) (7a) and d-2,3-dideoxy-
2,3-di¯uoro-myo-inositol 1,4,5,6-tetrakis(dibenzyl
phosphate)
(7b).ÐCompounds
5a
(85 mg,
0.156 mmol) or 5b (69 mg, 0.127 mmol) were
hydrogenated to give tetraols 6a or 6b. Without
puri®cation these were phosphorylated using
dibenzyl N,N-diisopropylphosphoramidite (5 eq)
to give the protected phosphates as oils, 7a
(141 mg, 74%) or 7b (79 mg, 51%). Enantiomer 7a;
Biological methods.ÐThe electrophysiological
methods evaluating the synergistic inhibition of
CaMK II-activated chloride current in T84 colonic
epithelial cells by 2a in the presence of 1a was as
previously described [20].
1
[ꢄ]d +2.4‹0.6 (c 0.0139 g/mL, CHCl3); H NMR
data (CDCl3): ꢅ 4.15±4.35 (1 H, m, inositol H),
4.4±4.65 (1 H, m, inositol H), 4.85±5.1 (19 H, m,
8ÂCH2Ph and 3Âinositol H), 5.35 (1 H, br dd,
3
2JHF 51.1, JHF 8.9, 2-H), 7.1±7.4 (40 H, m, Ph);
Acknowledgements
31P NMR data (CDCl3, referenced to 85% phos-
1
phoric acid, H decoupled): ꢅ 1.74 (s), 1.63 (s),
We thank the University of Manchester
Research Support Fund (K.R.H.S.). The mass
spectra were recorded at the Department of
Chemistry, University of Manchester.
0.88 (s), 0.72 (s); MS data (+ve electrospray):
1225 (M+H+, 5%), 1242 (M+H2O+, 100), 1247
(M+Na+, 27). Enantiomer 7b; [ꢄ]d 2.8‹0.5 (c
0.0145 g/mL, CHCl3).
Sodium salts of d-1,2-dideoxy-1,2-di¯uoro-myo-
inositol 3,4,5,6-tetrakisphosphate (2a) and d-2,3-
dideoxy-2,3-di¯uoro-myo-inositol 1,4,5,6-tꢀetrakis-
phosphate (2b).ÐEnantiomer 2a; mp>300 C; [ꢄ]d
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