Synthesis and membranoprotective properties of new disulfides with monoterpene and carbohydrate fragments

Article abstract of DOI:10.1007/s11172-015-0926-2

A cooxidation of carbohydrate and terpene thiols gives mixtures of disulfides containing 51 - 90% of the unsymmetric product. Membranoprotective and antioxidant properties of obtained unsymmetric and symmetric disulfides were evaluated based on their ability to inhibit the H₂O₂-induced hemolysis of erythrocytes, as well as the accumulation of secondary products of the peroxy oxidation of lipids and the oxidation of hemoglobin.

Full text of DOI:10.1007/s11172-015-0926-2

Russian Chemical Bulletin, International Edition, Vol. 64, No. 3, pp. 723—731, March, 2015
723
Synthesis and membranoprotective properties of new disulfides
with monoterpene and carbohydrate fragments*
S. V. Pestova,a E. S. Izmest´ev,a O. G. Shevchenko,b S. A. Rubtsova,a and A. V. Kuchina
aInstitute of Chemistry, Komi Scientific Center of the Ural Branch of the Russian Academy of Sciences,
48 ul. Pervomaiskaya, 167982 Syktyvkar, Russian Federation.
Fax: +7 (821 2) 21 8477. Eꢀmail: pestovaꢀsv@chemi.komisc.ru
bInstitute of Biology, Komi Scientific Center of the Ural Branch of the Russian Academy of Sciences,
28 ul. Kommunisticheskaya, 167982 Syktyvkar, Russian Federation
A cooxidation of carbohydrate and terpene thiols gives mixtures of disulfides containing
51—90% of the unsymmetric product. Membranoprotective and antioxidant properties of obꢀ
tained unsymmetric and symmetric disulfides were evaluated based on their ability to inhibit the
H2O2ꢀinduced hemolysis of erythrocytes, as well as the accumulation of secondary products of
the peroxy oxidation of lipids and the oxidation of hemoglobin.
Key words: disulfides, monoterpenes, thiols, monosaccharides, membranoprotective propꢀ
erties, erythrocytes, oxidation hemolysis, antioxidants.
The synthesis of new efficient and lowꢀtoxic sulfurꢀ
containing antioxidants based on natural substrates is of
significant interest for pharmacology, since the agents
based on natural raw materials, as a rule, are less dangerꢀ
ous for living organisms and environment. Glutathione,1,2
cystine,3 and lypoic acid4 are the commonly known enꢀ
dogenic sulfurꢀcontaining compounds with pronounced
antioxidant activity. In cells, these compounds are in
a labile equilibrium of the oxidized and the reduced forms
(—SS—/—SH), with the reduced form only being active.5
The transformation of the oxidized forms to the reduced
ones takes place upon treatment with highly specific
enzymes, for example, glutathione reductase and cystine
reductase, due to which a constant redox potential of the
cell is maintained.6 At the same time, disulfides also can
exhibit high antioxidant activity, in particular, such an
activity was described for diallyl disulfide encountered with
in plants of the genus Allium and formed from allicin thioꢀ
sulfinate and/or its precursors.7
thiosulfinates are prone to the disproportionation reacꢀ
tions, leading to the starting disulfide and the correspondꢀ
ing thiosulfonate, which can be subjected to oxidation, as
well. The formation of the starting disulfides from thiosulꢀ
finates proceeds spontaneously without involvement of
enzymes.9
The disulfides containing monosaccharide and monoꢀ
terpenoid fragments seem promising for pharmacology,
since they can possess not only an increased antioxidant
activity due to the presence of a terpene fragment,10 but
also relatively high bioaccessibility and biocompatibility
due to the presence of a carbohydrate fragment.11
The purpose of the present work is the synthesis and
primary screening of biological activity of symmetric and
unsymmetric disulfides containing terpene and/or carboꢀ
hydrate fragments in different combinations. The studies
also included evaluation of the contribution of each of the
fragments to the antioxidant and membranoprotective acꢀ
tivity of the conjugates obtained.
Disulfides can be involved in the oxidation reactions
with many active oxidants due to the presence of two reacꢀ
tion centers capable of binding with four and more equivaꢀ
lents of an oxidant. An increase in the amount of the
oxidant can lead to a stepwise formation of products with
the growing oxidation state of sulfur: thiosulfinates—thioꢀ
sulfonates—sulfinylsulfonates—disulfones, however, the
instability of thiosulfinates formed in the first step interꢀ
feres with further oxidation reactions.8 It is known that
Results and Discussion
In the present work, the target disulfides were obtained
from 6ꢀthiodiisopropylidenegalactopyranose and 1ꢀthioꢀ
diisopropylidenefructopyranose as the carbohydrate comꢀ
ponent and neomenthanethiol, isobornanethiol, cisꢀmyrꢀ
tanethiol, transꢀverbenethiol, and myrtenethiol as the terꢀ
pene component. The cooxidation of two different thiols
with iodine proceeded nonselectively with the formation
of two symmetric and one unsymmetric (containing a terꢀ
pene and a carbohydrate fragment) disulfides.
* Dedicated to Academician of the Russian Academy of Sciences
V. I. Minkin on the occasion of his 80th birthday.
Published in Russian in Izvestiya Akademii Nauk. Seriya Khimicheskaya, No. 3, pp. 0723—0731, March, 2015.
1066ꢀ5285/15/6403ꢀ0723 © 2015 Springer Science+Business Media, Inc.

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Pestova et al.
The starting 6ꢀthiodiisopropylidenegalactopyranose (1)
synthesized by the reaction of thioacetic acid and transꢀ
verbenol in the presence of a ZnCl2 catalyst.15
and 1ꢀthiodiisopropylidenefructopyranose (2) were obꢀ
tained from Dꢀgalactose 3 and Dꢀfructose 4 (Scheme 1).
Their reaction with acetone gave rise to the corresponding
diisopropylidenepyranoses 5 and 6; a preserved OH group
at positions 6 and 1, respectively, was used for further
transformations. The substitution for the OH group with
a SH group was carried out in three stages through the step
of the formation of thioacetates 7 and 8. The thioacetates
were obtained from the corresponding tosylate 9 and
iodide 10 (see Scheme 1).
The terpene thiols were synthesized using known proꢀ
cedures based on the substitution for the pꢀtoluenesulfoꢀ
nyl group in the corresponding tosylates with the thioaceꢀ
tic acid moiety12 (thiols 11a,b), or on the addition of thioꢀ
acetic acid at the double bond of the monoterpenes13
(thiol 11c), or on the reaction of Nꢀsulfinylbenzeneꢀ
sulfonamide with monoterpenes14 (thiol 11d) with subseꢀ
quent reduction of the formed thioacetates and sulfinylꢀ
sulfonamides. transꢀVerbenethiol (11e) was obtained by
the reduction of the corresponding thioacetate, in turn,
The oxidation of a mixture of two different thiols with
iodine (~20 C, a complete conversion) is a nonselective
process, which gives all the possible disulfides, however,
the rate of their formation is not the same and is deterꢀ
mined by steric effects, which facilitate or, vice versa,
hinder the coupling of two sulfanyl fragments. Unsymꢀ
metric disulfides 12a—e, 13a—e were found to be the maꢀ
jor products (Scheme 2). The largest amount of them was
formed in the oxidation of a mixture of thiol 2 and isoborꢀ
nanethiol 11b — up to 90% calculated on the total amount
of products, whereas the smallest amount was formed in
the case of thiol 2 and neomenthanethiol 11a (51%). Symꢀ
metric disulfides 14a—e based on terpene thiols were
formed in lower amounts: from 7 to 41% calculated on the
total amount of reaction products. In all the cases,
the formation of symmetric disulfides 15 and 16 with
monosaccharide fragments did not exceed 13% (Table 1).
All the disulfides were isolated by column chromatoꢀ
graphy in the individual state, their structure was estabꢀ
Scheme 1
BzIm is the benzimidazole. Py is the pyridine.

Hexapyranosyl terpenyl disulfides
Russ.Chem.Bull., Int.Ed., Vol. 64, No. 3, March, 2015
725
Scheme 2
lished by IR and NMR spectroscopy, as well as by eleꢀ
mental analysis.
conditions induced by the addition of hydrogen peroxide.
The membranoprotective activity of compounds with the
galactopyranose fragment decreases in the following order
(Fig. 1): 12d = 12c > 12a = 12b > 15 = standard (diꢀnꢀ
propyl disulfide) > 12e. This means that the conjugates
with cisꢀmyrtanyl, myrtenyl, neomenthyl, and isobornyl
moieties are the most active. The lowest activity was obꢀ
served for the disulfide with transꢀverbenyl fragment.
The structurally similar compounds 13a—e containing
a fructopyranose moiety exhibited a lower activity as comꢀ
pared to disulfides with galactopyranose fragment (Fig. 2).
The calculations showed that the replacement of the
galactopyranose fragment with a fructopyranose one led
to a 1.2—1.6 times decrease in the membranoprotective
activity for most compounds, except a lowꢀactive sulfide
12e containing a transꢀverbenyl fragment (see Figs 1
and 2). Thus, in the case when erythrocytes were protectꢀ
ed by the conjugates with a galactopyranose fragment, the
value of relative hemolysis was 44—69%, whereas when
similar compounds with a fructopyranose fragment were
used, it was 69—91%. However, among the disulfides with
a fructopyranose moiety the compounds containing myrteꢀ
nyl, cisꢀmyrtanyl, or neomenthyl fragments were found to
be the most active. The regressive analysis indicates the
presence of a statistically significant close positive relaꢀ
tionship between the activities of the corresponding galacꢀ
toꢀ and fructosoꢀcontaining disulfides with five different
terpene fragments in the composition (RS = 0.9, p = 0.037).
For the symmetric disulfides containing no sugar fragꢀ
ments (Fig. 3), the highest membranoprotective activity
was found for compounds containing cisꢀmyrtanyl (14d),
myrtenyl (14c), and isobornyl (14b) moieties. The lowest
activity was found in the disulfide with transꢀverbenyl fragꢀ
ment (14e). For the symmetric neomenthaneꢀderived diꢀ
sulfide (14a), a statistically significant membranoprotecꢀ
tive activity was not found (no data reported).
The screening of biological activity of disulfides was
carried out in the in vitro system using blood erythrocytes
of laboratory animals.16—21 Since the toxicity of disulfides
can impose considerable limitations on their further use,
the compounds were first evaluated on their hemolytic
activity.
Among all the conjugates under study, the hemolytic
activity was found only in unsymmetric disulfides conꢀ
taining neomenthyl (12a, 13a) or isobornyl (13b) fragꢀ
ments. The addition of these compounds to a suspension
of erythrocytes in the concentration of 10 mol L–1 caused
a 2.1—3.4 times excess of a spontaneous hemolysis level.
Most of the tested compounds showed an ability to
inhibit the death of erythrocytes under the acute stress
Table 1. Oxidation of mixtures of terpene and carboꢀ
hydrate thiols
Mixture
of thiols
Products
Proportion of corresꢀ
ponding disulfide* (%)
1 + 11a
1 + 11b
1 + 11c
1 + 11d
1 + 11e
2 + 11a
2 + 11b
2 + 11c
2 + 11d
2 + 11e
12a + 14a + 15
12b + 14b + 15
12c + 14c + 15
12d + 14d + 15
12e + 14e + 15
13a + 14a + 16
13b + 14b + 16
13c + 14c + 16
13d + 14d + 16
13e + 14e + 16
53 : 34 : 13
75 : 19 : 6
71 : 24 : 5
64 : 29 : 7
78 : 18 : 4
51 : 41 : 8
90 : 7 : 3
72 : 20 : 8
62 : 30 : 8
83 : 10 : 7
* Calculated from the ratio of integral intensities of nonꢀ
overlapping signals for the protons in the NMR spectra
of the reaction mixture.

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Pestova et al.
Hemolysis (%)
70
1 h
2 h
3 h
4 h
5 h
60
50
40
30
20
10
Control
12a
12b
12e
12d
12c
15
Standard
Fig. 1. Influence of compounds 12a—e, 15 and Prn2S2 (a standard) in the concentration of 10 mol L–1 on the degree of the
H2O2ꢀinduced hemolysis of erythrocytes after 1—5 h of incubation.
Hemolysis (%)
80
1 h
2 h
3 h
4 h
5 h
70
60
50
40
30
20
10
Control
13a
13b
13e
13d
13c
16
Fig. 2. Influence compounds 13a—e, 16 in the concentration of 10 mol L–1 on the degree of the H2O2ꢀinduced hemolysis of
erythrocytes after 1—5 h of incubation.
Hemolysis (%)
70
1 h
2 h
3 h
4 h
5 h
60
50
40
30
20
10
Control
14b
14e
14d
14c
Standard
Fig. 3. Influence compounds 14b—e and Prn2S2 (standard) in the concentration of 10 mol L–1 on the degree of the H2O2ꢀinduced
hemolysis of erythrocytes after 1—5 h of incubation.

Hexapyranosyl terpenyl disulfides
Russ.Chem.Bull., Int.Ed., Vol. 64, No. 3, March, 2015
727
Level of hemolysis
1.0
of secondary products of POL and to protect cells from
damage.
Finally, most disulfides obtained in this work (except
those containing a transꢀverbenyl fragment) exhibited the
ability to inhibit transformation of oxyhemoglobin of
erythrocytes to methemoglobin upon treatment with hyꢀ
drogen peroxide, which also indicates the presence of
a pronounced antioxidant activity. The highest activity
was found in the conjugates with the myrtenyl/cisꢀmyrtaꢀ
nyl fragments (independent of the structure of the second
substituent). The preincubation of erythrocytes with these
disulfides led to a 2—2.4 times decrease in the level of
accumulation of methemoglobin as compared to the conꢀ
trol samples (containing no compounds under study),
whereas a comparison agent inhibited the formation of
methemoglobin only by 1.4 times.
In conclusion, we for the first time synthesized symꢀ
metric and unsymmetric disulfides containing five terpene
and two carbohydrate moieties in different combinations.
A biological model system was used to show that the antiꢀ
oxidant and membranoprotective activity of the conjuꢀ
gates obtained depended on the structure of both substituꢀ
ents (the terpene and the carbohydrate fragments). The
most active are disulfides, which have cisꢀmyrtanyl, myrteꢀ
nyl, and neomenthyl fragments in the composition, with
the activity of compounds with a galactopyranose fragꢀ
ment being slightly higher than that of similar structures
with a fructopyranose fragment. For symmetric disulfides
containing no sugar moieties, the highest activity was
found for compounds containing cisꢀmyrtanyl, myrtenyl,
and isobornyl fragments. A low activity in this model sysꢀ
tem was found for disulfides of different structure containꢀ
ing transꢀverbenyl fragment.
0.8
0.6
0.4
0.2
12a—14a 12b—14b 12c—14c 12d—14d 12e—14e
Fig. 4. Relative membranoprotective activity (the ratio of the
hemolysis level in the tested and control samples 5 h after addiꢀ
tion of H2O2) of compounds 12a—e—14a—e in the concentraꢀ
tion of 10 mol L–1
.
A comparative evaluation of the membranoprotective
activity showed (Fig. 4) that for most disulfides it depends
on the structure of both the terpene and the carbohydrate
fragments. Only for the conjugates with a transꢀverbenyl
moiety, the presence and the character of the carbohydrate
part of the molecule have no influence on the biological
activity of the compounds. Generally, it can be stated that
the highest ability to protect cells from the influence of
hydrogen peroxide is possessed by compounds containing
cisꢀmyrtanyl/myrtenyl and galactose fragments.
An ability of compounds to inhibit the process of perꢀ
oxide oxidation of lipids (POL) in erythrocytes under conꢀ
ditions of the acute oxidation stress can be evaluated based
on the data given in Fig. 5.
The disulfides with the cisꢀmyrtanyl/myrtenyl fragꢀ
ments in a combintion with the galactose moiety inhibited
accumulation of secondary products of POL most activeꢀ
ly. A comparison reference and symmetric disulfides with
two carbohydrate fragments showed relatively low activity
in this model system, inhibiting POL to 72—80% from the
control data.
The found relationships between the structure and the
biological activity of disulfides with terpene and/or carboꢀ
hydrate moieties in the composition allow us to carry out
a purposeful synthesis of new biologically active compounds,
promising for pharmacology and food production.
All seventeen conjugates studied are characterized by
the statistically significant positive dependence (RS = 0.56,
p = 0.020) between the ability to inhibit the accumulation
Experimental
Antioxidant activity
1.0
IR spectra were recorded on a Shimadzu IR Prestige 21
Fourierꢀtransform IR spectrometer for neat samples or in KBr
pellets. Melting points were determined on a GallencampꢀSanyo
apparatus. 1H and 13C NMR spectra were recorded on a Bruker
Avanceꢀ300 spectrometer (300.17 (1H) and 75.48 MHz (13C)) in
solutions in CDCl3, using residual signals of chloroform as
a reference. The 1H and 13C signals were completely assigned
using 2D homoꢀ (1H—1H COSY, 1H—1H NOESY) and heteroꢀ
0.8
0.6
0.4
0.2
1
nuclear experiments (1H—13C HSQC, H—13C HMBC). Optiꢀ
cal rotation angles were measured on a Kruss P3002RS automatꢀ
ed digital polarimeter. Thinꢀlayer chromatography was carried
out on Sorbfil plates in the Et2O—benzene, CHCl3—Et2O, and
light petroleum—AcOEt solvent systems in different gradient
ratio; solutions of phosphomolybdic acid and KMnO4 were used
for visualization. Elemental anlysis was carried out on an EA
12a—14a 12b—14b 12c—14c 12d—14d 12e—14e
Fig. 5. Relative antioxidant activity (the ratio of the content of
2ꢀthiobarbituric acid in in the tested and control samples 5 h
after addition of H2O2) of compounds 12a—e—14a—e in the
concentration of 10 mol L–1
.

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Pestova et al.
1110 CHNSꢀO automated analyzer. All the reactions were carꢀ
ried out with freshly distilled solvents. Column chromatography
was performed on Alfa Aesar silica gel (0.06—0.2 mm) in the
same solvent systems as for TLC.
1,2:3,4ꢀDiꢀOꢀisopropylideneꢀ6ꢀthioacetylꢀ6ꢀdeoxyꢀꢀDꢀgaꢀ
lactopyranose (7) was obtained according to the procedure.28
22
The yield was 77%, []D –15.6 (c 0.30, CHCl3) (cf. Ref. 28:
the yield 99%, []D22 –15.0 (c 1.8, CHCl3)).
(1S,2S,5R)ꢀ2ꢀIsopropylꢀ5ꢀmethylcyclohexanethiol (neomenꢀ
thanethiol (11a)) was obtained according to the procedure22 from
2,3:4,5ꢀDiꢀOꢀisopropylideneꢀꢀDꢀfructopyranose (6) was synꢀ
thesized according to the procedure.26 The yield was 85%, []D
22
25
Lꢀ(–)ꢀmenthol (99%) made by Alfa Aesar ([]D –50.0, c 0.1,
–45.6 (c 0.35, Me2CO) (cf. Ref. 26: the yield 90%, []D22 –46.0
(c 0.36, Me2CO)).
2,3:4,5ꢀDiꢀOꢀisopropylideneꢀ1ꢀiodoꢀ1ꢀdeoxyꢀꢀDꢀfructopyrꢀ
anose (10) was obtained according to the procedure.29 The yield
was 64%, []D22 –47.6 (c 0.88, CHCl3).
2,3:4,5ꢀDiꢀOꢀisopropylideneꢀ1ꢀthioacetylꢀ1ꢀdeoxyꢀꢀDꢀfrucꢀ
topyranose (8) was synthesized according to the procedure.30
The yield was 72%, []D22 –3.1 (c 0.95, Me2CO) (cf. Ref. 30: the
yield 76%, []D22 –2.9 (c 1.0, CHCl3)).
22
EtOH). A colorless liquid. The yield was 58%, []D +53.2
22
(c 0.83, EtOH) (cf. Ref. 22: the yield 60%, []D 53.0 (c 0.8,
EtOH)).
(1S,2S,4S)ꢀ1,7,7ꢀTrimethylbicyclo[2.2.1]heptaneꢀ2ꢀthiol
(isobornanethiol (11b)) was synthesized according to the proꢀ
cedure23 from (–)ꢀborneol (98%) made by Alfa Aesar
22
([]D –36.0, c 5.0, EtOH). A white powder. The yield was
22
83%, []D +46.5 (c 0.1, EtOH) (cf. Ref. 23: the yield 85%,
[]D22 +47.0 (c 0.1, EtOH)).
1,2:3,4ꢀDiꢀOꢀisopropylideneꢀ6ꢀmercaptoꢀ6ꢀdeoxyꢀꢀDꢀgaꢀ
lactopyranose (1) was obtained according to the procedure.31
A dense yellow liquid. The yield was 91% (benzene—Et2O, 10 : 1),
(1R,2S,5R)ꢀ(6,6ꢀDimethylbicyclo[3.1.1]heptꢀ2ꢀyl)methaneꢀ
thiol (cisꢀmyrtanethiol (11c)) was obtained through thioacetate
by a direct addition of thioacetic acid to (–)ꢀꢀpinene according
to the described procedure.24 A colorless liquid. The yield was
22
[]D –70 (c 0.47, CHCl3) (cf. Ref. 31: the yield 70%,
[]D22 –88.7 (CHCl3)).
22
71%, []D –45.6 (c 0.35, CHCl3) (cf. Ref. 24: the yield 37%,
2,3:4,5ꢀDiꢀOꢀisopropylideneꢀ1ꢀmercaptoꢀ1ꢀdeoxyꢀꢀDꢀfrucꢀ
topyranose (2) was obtained according to the procedure.32
A white powder. The yield was 94% (benzene—Et2O, 10 : 1),
[]D22 –44.0 (c 0.40, CHCl3) (cf. Ref. 32: the yield 65%, angle of
rotation not reported).
[]D22 –75.9 (c 5.18, CHCl3)).
(1S,5R)ꢀ(6,6ꢀDimethylbicyclo[3.1.1]heptꢀ2ꢀenꢀ2ꢀyl)methꢀ
anethiol (myrtenethiol (11d)) was synthesized according to the
procedure14 from (–)ꢀꢀpinene (99%) made by Alfa Aesar ([]D
25
–21.0, without solvent). A colorless liquid. The yield was 51%,
Oxidation of thiols with iodine (general procedure). A solution
of I2 (0.127 g, 0.5 mmol) in EtOH (6 mL) was added slowly to
a mixture containing a terpene thiol (1 mmol) and a thiol carboꢀ
hydrate (0.276 g, 1 mmol) in ethanol (4 mL) with vigorous stirꢀ
ring until a yellow color was persistent. The mixture was stirred
for 2 h, the solvent was evaporated in vacuo. The residue was
dissolved in CHCl3 (30 mL), and washed with saturated solution
of Na2S2O3 (2 mL). The organic layer was separated, dried
with anhydrous CaCl2, the solvent was evaporated in vacuo,
the residue was separated by column chromatography (eluent
CHCl3—Et2O, 5 : 1; visualization with KMnO4).
22
[]D –46.5 (c 0.1, EtOH) (cf. Ref. 14: the yield 84%,
[]D22 –42.6 (c 0.4, CHCl3)).
2,6,6ꢀTrimethylbicyclo[3.1.1]heptꢀ3ꢀenеꢀ2ꢀthiol (transꢀverꢀ
benethiol (11e)). The compounds MeC(O)SH (0.076 g, 1 mmol)
and ZnCl2 (0.014 g, 0.1 mmol) were sequentially added to transꢀ
25
verbenol25 (0.152 g, 1 mmol) ([]D +1.0, without solvent) in
CH2Cl2 (5 mL) with stirring. The mixture was stirred for
10—15 min, treated with water (20 mL), and extracted with
chloroform (3×10 mL). The combined organic layers were dried
with CaCl2, the solvent was evaporated in vacuo. The residue
was dissolved in Et2O (20 mL) and LiAlH4 (0.019 g, 0.5 mmol)
was added under argon. After 15 min, water (10 mL) was added
slowly, extracted with light petroleum (3×15 mL), dried with
CaCl2, the solvent was evaporated in vacuo. The residue was
separated by column chromatography (eluent light petroleum,
visualization with KMnO4). A colorless liquid. The yield was
90%. Found (%): C, 71.42; H, 9.48; S, 19.10. C10H16S. Calcuꢀ
lated (%): C, 71.37; H, 9.58; S, 19.05. IR, /cm–1: 675 (C—S),
820, 1372, 1645 (C=C), 2924. 1H NMR (CDCl3), : 0.90 (s, 3 H,
9´ꢀMe); 1.35 (s, 3 H, 10´ꢀMe); 1.31—1.43 (m, 1 H, HA(7´));
1.63 (d, 1 H, SH, J = 8.5 Hz); 1.72 (s, 3 H, 8´ꢀMe); 2.03 (t, 1 H,
H(5´), J = 5.4 Hz); 2.17—2.25 (m, 1 H, H(1´)); 2.35 (dt, 1 H,
HB(7´), J = 9.1 Hz, J = 5.6 Hz); 3.65—3.75 (m, 1 H, H(4´));
5.26—5.32 (m, 1 H, H(3´)). 13C NMR, : 20.37 (C(9´)),
22.63 (C(8´)), 26.52 (C(10´)), 28.29 (C(7´)), 40.81 (C(1´)),
42.82 (C(6´)), 47.35 (C(5´)), 49.54 (C(4´)), 119.82 (C(3´)),
145.69 (C(2´)).
6ꢀ{[(1S,2S,5R)ꢀ2ꢀIsopropylꢀ5ꢀmethylcyclohexyl]disulfanyl}ꢀ
1,2:3,4ꢀdiꢀOꢀisopropylideneꢀ6ꢀdeoxyꢀꢀDꢀgalactopyranose (12a).
A dense yellow liquid. The yield was 53% (0.125 g), []D22 +33.0
(c 0.71, CHCl3). Found (%): C, 59.02; H, 8.66; S, 14.24.
C22H38O5S2. Calculated (%): C, 59.16; H, 8.58; S, 14.36. IR,
/cm–1: 646 (C—S), 999, 1071 (C—O), 1169 (O—C—O), 1211,
1
2918. H NMR (CDCl3), : 0.86—0.92 (m, 1 H, He(4´)); 0.91
(d, 3 H, 7´ꢀMe, J = 6.6 Hz); 0.93 (d, 3 H, 10´ꢀMe, J = 6.6 Hz);
1.01 (d, 3 H, 9´ꢀMe, J = 6.6 Hz; 1.10—1.33 (m, 3 H, H(2´),
He(6´), He(3´)); 1.37 (s, 3 H, 8ꢀMe); 1.38 (s, 3 H, 12ꢀMe); 1.47
(s, 3 H, 11ꢀMe); 1.62 (s, 3 H, 9ꢀMe); 1.58—1.81 (m, 3 H, He(4´),
H(8´), Ha(3´)); 1.81—2.00 (m, 1 H, H(5´)); 2.25—2.35 (m, 1 H,
Ha(6´)); 2.90 (dd, 2 H, H(6), J = 6.7 Hz, J = 2.1 Hz); 3.36—3.43
(m, 1 H, H(1)); 4.11—4.18 (m, 1 H, H(5)); 4.30—4.39 (m, 2 H,
H(2), H(3)); 4.67 (dd, 1 H, H(4), J = 8.0 Hz, J = 2.2 Hz); 5.57
(d, 1 H, H(1), J = 5.0 Hz). 13C NMR, : 20.74 (C(10´)), 21.20
(C(9´)), 22.11 (C(7´)), 24.47 (C(12)), 25.04 (C(8)), 25.91 (C(3´)),
25.98 (C(11), C(5´)), 26.25 (C(9)), 29.99 (C(8´)), 35.38 (C(4´)),
38.30 (C(6)), 40.12 (C(6´)), 48.80 (C(2´)), 53.70 (C(1´)), 66.67
(C(5)), 70.64 (C(2)), 70.98 (C(4)), 71.70 (C(3)), 96.67 (C(1)),
108.76 (C(7)), 109.30 (C(10)).
1,2:3,4ꢀDiꢀOꢀisopropylideneꢀꢀDꢀgalactopyranose (5) was
obtained according to the procedure.26 The yield was 85%, []D
22
–45.6 (c 0.35, Me2CO) (cf. Ref. 26: the yield 72—90%,
[]D22 –46.0 (c 0.36, Me2CO)).
1,2:3,4ꢀDiꢀOꢀisopropylideneꢀ6ꢀOꢀ(pꢀtoluenesulfonyl)ꢀꢀDꢀ
galactopyranose (9) was synthesized according to the proceꢀ
1ꢀ{[(1S,2S,5R)ꢀ2ꢀIsopropylꢀ5ꢀmethylcyclohexyl]disulfanyl}ꢀ
2,3;4,5ꢀdiꢀOꢀisopropylideneꢀ1ꢀdeoxyꢀꢀDꢀfructopyranose (13a).
A yellow powder. M.p. 53—54 C. The yield was 51% (0.120 g),
dure.27 The yield was 98%, []D –44.6 (c 0.30, CHCl3)
22
(cf. Ref. 27: the yield 87%, []D22 –44.0 (c 0.28, CHCl3)).
[]D +99.0 (c 0.58, CHCl3). Found (%): C, 59.08; H, 8.49;
22

Hexapyranosyl terpenyl disulfides
Russ.Chem.Bull., Int.Ed., Vol. 64, No. 3, March, 2015
729
S, 14.30. C22H38O5S2. Calculated (%): C, 59.16; H, 8.58;
S, 14.36. IR, /cm–1: 511 (S—S), 642 (C—S), 988, 1067 (C—O),
1165 (O—C—O), 2918. 1H NMR (CDCl3), : 0.89 (d, 3 H,
7´ꢀMe, J = 6.3 Hz); 0.91 (d, 3 H, 9´ꢀMe, J = 6.3 Hz); 0.85—0.95
(m, 1 H, He(4´)); 0.99 (d, 3 H, 10´ꢀMe, J = 6.6 Hz); 1.06—1.30
(m, 3 H, H(2´), He(6´), He(3´)); 1.36 (s, 3 H, 12ꢀMe); 1.48 (s, 3 H,
8ꢀMe); 1.50 (s, 3 H, 11ꢀMe); 1.55 (s, 3 H, 9ꢀMe); 1.55—1.80
(m, 3 H, Ha(3´), Ha(4´), H(8´)); 1.80—1.98 (m, 1 H, H(5´));
2.24—2.36 (m, 1 H, Ha(6´)); 3.10 (d, 1 H, HA(1), J = 13.8 Hz);
3.31—3.42 (m, 2 H, HB(1), H(1´)); 3.76 (d, 1 H, HA(6),
J = 12.9 Hz); 3.93 (d, 1 H, HB(6), J = 12.7 Hz); 4.24 (d, 1 H,
H(5), J = 7.7 Hz); 4.39 (d, 1 H, H(3), J = 2.5 Hz); 4.63 (dd, 1 H,
H(4), J = 8.0 Hz, J = 2.5 Hz). 13C NMR, : 20.84 (C(10´)),
21.18 (C(9´)), 22.15 (C(7´)), 24.10 (C(12)), 25.76 (C(8)), 25.87
(C(3´)), 25.93 (C(11)), 26.18 (C(5´)), 26.64 (C(9)), 26.89 (C(8´)),
35.41 (C(4´)), 39.91 (C(6´)), 48.84 (C(2´)), 49.06 (C(1)), 54.16
(C(1´)), 61.68 (C(6)), 70.39 (C(4)), 70.82 (C(5)), 71.96 (C(3)),
102.93 (C(2)), 108.66 (C(7)), 109.08 (C(10)).
lactopyranose (12c). A dense yellow liquid. The yield was 62%
(0.195 g), []D22 –113.0 (c 0.64, CHCl3). Found (%): C, 59.51;
H, 8.24; S, 14.31. C22H36O5S2. Calculated (%): C, 59.43;
H, 8.16; S, 14.42. IR, /cm–1: 511 (S—S), 646 (C—S), 1071
(C—O), 1169 (O—C—O), 2913. 1H NMR (CDCl3), : 0.83—1.04
(m, 1 H, HA(6´)); 1.02 (s, 3 H, 9´ꢀMe); 1.22 (s, 3 H, 8´ꢀMe);
1.37 (s, 3 H, 8ꢀMe); 1.38 (s, 3 H, 12ꢀMe); 1.48 (s, 3 H, 11ꢀMe);
1.48—1.64 (m, 1 H, HA(6´)); 1.61 (s, 3 H, 9ꢀMe); 1.75—2.16
(m, 3 H, H(1´), H(2´), HB(4´)); 2.27—2.53 (m, 2 H, H(5´), HB(6´));
2.64—3.02 (m, 6 H, H(3´), H(10´), H(6)); 4.08—4.18 (m, 1 H,
H(5)); 4.32—4.45 (m, 2 H, H(2), H(3)); 4.67 (dd, 1 H, H(4),
J = 8.0 Hz, J = 2.2 Hz); 5.57 (d, 1 H, H(1), J = 5.0 Hz). 13C NMR,
: 21.81 (C(4´)), 23.31 (C(9´)), 24.48 (C(12)), 25.03 (C(8)), 26.00
(C(11)), 26.08 (C(9)), 27.93 (C(8´)), 33.25 (C(6´)), 38.04 (C(6)),
38.69 (C(7´)), 40.21 (C(5´)), 41.25 (C(1´)), 45.44 (C(3´)), 45.92
(C(10´)), 66.57 (C(5)), 70.61 (C(2)), 70.94 (C(4)), 71.50 (C(3)),
96.71 (C(1)), 108.79 (C(7)), 109.32 (C(10)).
1ꢀ[(cis)ꢀ(1R,2R,5R)ꢀ6,6ꢀDimethylbicyclo[3.1.1]heptꢀ2ꢀylꢀ
methyldisulfanyl]ꢀ2,3;4,5ꢀdiꢀOꢀisopropylideneꢀ1ꢀdeoxyꢀꢀDꢀfructoꢀ
pyranose (13c). A dense yellow liquid. The yield was 60% (0.192 g),
1,2:3,4ꢀDiꢀOꢀisopropylideneꢀ6ꢀ{[(1S,2S,4S)ꢀ1,7,7ꢀtrimethꢀ
ylbicyclo[2.2.1]heptꢀ2ꢀyl]disulfanyl}ꢀ6ꢀdeoxyꢀꢀDꢀgalactopyrꢀ
anose (12b). A dense colorless liquid. The yield was 73% (0.243 g),
22
[]D –47.0 (c 0.32, CHCl3). Found (%): C, 59.60; H, 8.09;
22
[]D –106.0 (c 0.22, CHCl3). Found (%): C, 59.49; H, 8.22;
S, 14.51. C22H36O5S2. Calculated (%): C, 59.43; H, 8.16; S, 14.42.
S, 14.49. C22H36O5S2. Calculated (%): C, 59.43; H, 8.16; S, 14.42.
IR, /cm–1: 516 (S—S), 578 (C—S), 1065 (C—O), 1165 (O—C—O),
IR, /cm–1: 511 (S—S), 646 (C—S), 1071 (C—O), 1140, 1169
2913. H NMR (CDCl3), : 0.84—1.07 (m, 1 H, HA(7´)); 1.01
1
1
(O—C—O), 2953. H NMR (CDCl3), : 0.84 (s, 3 H, 8´ꢀMe);
(s, 3 H, 9´ꢀMe); 1.21 (s, 3 H, 8´ꢀMe); 1.38 (s, 3 H, 12ꢀMe); 1.47
(s, 3 H, 8ꢀMe); 1.53 (s, 3 H, 11ꢀMe); 1.56 (s, 3 H, 9ꢀMe); 1.45—1.62
(m, 1 H, HA(4´)); 1.67—2.17 (m, 5 H, H(1´), H(2´), HB(4´),
H(3´)); 2.27—2.52 (m, 2 H, H(5´), HB(7´)); 2.79—2.95 (m, 2 H,
H(10´)); 3.09 (d, 1 H, HA(1), J = 13.8 Hz); 3.36—3.45 (m, 3 H,
HB(1)); 3.79 (d, 1 H, HA(6), J = 13.0 Hz); 3.94 (dd, 1 H, HB(6),
J = 12.9 Hz, J = 1.4 Hz); 4.26 (d, 1 H, H(5), J = 8.0 Hz); 4.33
(d, 1 H, H(3), J = 2.5 Hz); 4.63 (dd, 1 H, H(4), J = 8.0 Hz,
J = 2.5 Hz)). 13C NMR, : 20.17 (C(4´)), 21.83 (C(9´)), 24.13
(C(12)), 25.58 (C(8)), 25.94 (C(11)), 26.59 (C(9)), 26.00 (C(11)),
26.08 (C(9)), 26.10 (C(3´)), 27.94 (C(8´)), 33.25 (C(7´)), 38.69
(C(6´)), 40.12 (C(5´)), 41.26 (C(1´)), 45.58 (C(2´)), 46.37
(C(10´)), 49.69 (C(1)), 61.62 (C(6)), 70.41 (C(4)), 70.80 (C(5)),
72.24 (C(3)), 102.66 (C(2)), 108.60 (C(7)), 109.13 (C(10)).
1,2:3,4ꢀDiꢀOꢀisopropylideneꢀ6ꢀ[(1S,5R)ꢀ6,6ꢀdimethylbiꢀ
cyclo[3.1.1]heptꢀ2ꢀenꢀ2ꢀylmethyldisulfanyl]ꢀ6ꢀdeoxyꢀꢀDꢀgalacꢀ
topyranose (12d). A dense yellow liquid. The yield was 58%
(0.164 g), []D22 –105.0 (c 0.94, CHCl3). Found (%): C, 59.63;
H, 7.51; S, 14.43. C22H34O5S2. Calculated (%): C, 59.70;
H, 7.74; S, 14.49. IR, /cm–1: 511 (S—S), 679 (C—S), 1071
(C—O), 1169 (O—C—O), 1645 (C=C), 2918. 1H NMR (CDCl3),
: 0.85 (s, 3 H, 9´ꢀMe); 1.16—1.31 (m, 1 H, HA(4´)); 1.31 (s, 3 H,
8´ꢀMe); 1.37 (s, 3 H, 8ꢀMe); 1.38 (s, 3 H, 12ꢀMe); 1.47 (s, 3 H,
11ꢀMe); 1.61 (s, 3 H, 9ꢀMe); 2.05—2.15 (m, 3 H, H(5´)); 2.21
(t, 1 H, H(1´), J = 5.2 Hz); 2.25—2.33 (m, 2 H, H(7´)); 2.38—2.48
(m, 1 H, HB(4´)); 2.85—3.00 (m, 2 H, H(6)); 3.29—3.41 (m, 2 H,
H(10´)); 4.12 (t, 1 H, H(5), J = 6.6 Hz); 4.35 (dd, 1 H, H(2),
J = 5.0 Hz, J = 2.5 Hz); 4.41 (dd, 1 H, H(3), J = 8.0 Hz, J = 1.1 Hz);
4.66 (dd, 1 H, H(4), J = 7.8 Hz, J = 2.3 Hz); 5.50 (s, 1 H, H(3´));
5.57 (d, 1 H, H(1), J = 5.0 Hz). 13C NMR, : 21.29 (C(9´)),
24.47 (C(12)), 25.01 (C(8)), 25.98 (C(11)), 26.08 (C(9)), 26.16
(C(8´)), 31.43 (C(4´)), 31.74 (C(7´)), 37.95 (C(6)), 38.15 (C(6´)),
40.45 (C(5´)), 45.26 (C(1´)), 45.73 (C(10´)), 66.65 (C(5)), 70.63
(C(2)), 70.92 (C(4)), 71.36 (C(3)), 96.71 (C(1)), 108.74 (C(7)),
109.28 (C(10)), 121.63 (C(3´)), 142.83 (C(2´)).
0.92 (s, 3 H, 9´ꢀMe); 1.05 (s, 3 H, 10´ꢀMe); 1.10—1.33 (m, 2 H,
Hen(5´), Hen(6´)); 1.37 (s, 3 H, 8ꢀMe); 1.38 (s, 3 H, 12ꢀMe);
1.47 (s, 3 H, 11ꢀMe); 1.61 (s, 3 H, 9ꢀMe); 1.64—1.81 (m, 3 H,
Hex(5´), Hex(6´), H(4´)); 1.83—2.08 (m, 2 H, Hen(3´), Hex(3´));
2.85—3.08 (m, 3 H, H(2´), H(6)); 4.10—4.19 (m, 1 H, H(5));
4.35 (q, 1 H, H(2), J = 4.9 Hz, J = 2.6 Hz); 4.40 (dd, 1 H, H(3),
J = 7.8 Hz, J = 1.8 Hz); 4.66 (dd, 1 H, H(4), J = 7.8 Hz, J = 2.3 Hz);
5.57 (d, 1 H, H(1), J = 5.0 Hz). 13C NMR, : 14.02 (C(10´)),
20.00 (C(8´)), 20.48 (C(9´)), 24.46 (C(12)), 25.03 (C(8)), 25.99
(C(11)), 26.08 (C(9)), 27.31 (C(5´)), 38.25 (C(6´)), 38.33 (C(6)),
40.51 (C(3´)), 45.93 (C(4´)), 47.16 (C(1´)), 49.89 (C(7´)), 63.93
(C(2´)), 66.44 (C(5)), 70.66 (C(2)), 70.93 (C(4)), 71.30 (C(3)),
96.72 (C(1)), 108.76 (C(7)), 109.26 (C(10)).
2,3;4,5ꢀDiꢀOꢀisopropylideneꢀ1ꢀ[(1S,2S,4S)ꢀ1,7,7ꢀtrimethylꢀ
bicyclo[2.2.1]heptꢀ2ꢀyldisulfanyl]ꢀ1ꢀdeoxyꢀꢀDꢀfructopyranose
22
(13b). A dense yellow liquid. The yield was 86% (0.342 g), []D
–24.0 (c 0.15, CHCl3). Found (%): C, 59.49; H, 8.22; S, 14.49.
C22H36O5S2. Calculated (%): C, 59.43; H, 8.16; S, 14.42. IR,
/cm–1: 520 (S—S), 579 (C—S), 1069 (C—O), 1130, 1165
1
(O—C—O), 2949. H NMR (CDCl3), : 0.84 (s, 3 H, 8´ꢀMe);
0.91 (s, 3 H, 9´ꢀMe); 1.06 (s, 3 H, 10´ꢀMe); 1.09—1.32 (m, 2 H,
Hen(5´), Hen(6´)); 1.38 (s, 3 H, 12ꢀMe); 1.47 (s, 3 H, 8ꢀMe);
1.53 (s, 3 H, 11ꢀMe); 1.57 (s, 3 H, 9ꢀMe); 1.61—1.81 (m, 3 H,
Hex(5´), Hex(6´), H(4´)); 1.80—2.07 (m, 2 H, Hen(3´), Hex(3´));
3.06 (dd, 1 H, H(2´), J = 9.4 Hz, J = 6.1 Hz); 3.13 (d, 1 H,
HA(1), J = 13.8 Hz); 3.44 (d, 1 H, HB(1), J = 13.8 Hz); 3.79 (d, 1 H,
HA(6), J = 13.2 Hz); 3.94 (dd, 1 H, HB(6), J = 12.9 Hz,
J = 1.4 Hz); 4.26 (d, 1 H, H(5), J = 7.7 Hz); 4.36 (d, 1 H, H(3),
J = 2.5 Hz); 4.64 (dd, 1 H, H(3), J = 7.8 Hz, J = 2.6 Hz). 13C NMR,
: 14.07 (C(10´)), 20.00 (C(8´)), 20.48 (C(9´)), 24.12 (C(12)),
25.29 (C(8)), 25.69 (C(11)), 25.99 (C(9)), 27.38 (C(5´)), 38.30
(C(6´)), 40.35 (C(3´)), 45.88 (C(4´)), 47.16 (C(1´)), 49.78 (C(7´),
C(1)), 61.59 (C(6)), 63.96 (C(2´)), 70.40 (C(4)), 70.81 (C(5)),
72.11 (C(3)), 102.79 (C(2)), 108.59 (C(7)), 109.10 (C(10)).
6ꢀ[(cis)ꢀ(1R,2R,5R)ꢀ6,6ꢀDimethylbicyclo[3.1.1]heptꢀ2ꢀylꢀ
methyldisulfanyl]ꢀ1,2:3,4ꢀdiꢀOꢀisopropylideneꢀ6ꢀdeoxyꢀꢀDꢀgaꢀ
2,3;4,5ꢀDiꢀOꢀisopropylideneꢀ1ꢀ[(1S,5R)ꢀ6,6ꢀdimethylbicycloꢀ
[3.1.1]heptꢀ2ꢀenꢀ2ꢀylmethyldisulfanyl]ꢀ1ꢀdeoxyꢀꢀDꢀfructopyrꢀ

730
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Pestova et al.
anose (13d). A dense yellow liquid. The yield was 52% (0.142 g),
[]D –75.0 (c 0.51, CHCl3). Found (%): C, 59.60; H, 7.58;
Bis(1,2;3,4ꢀdiꢀOꢀisopropylideneꢀ6ꢀdeoxyꢀꢀDꢀgalactopyraꢀ
nosꢀ6ꢀyl) disulfide (15). A dense yellow liquid. The yield was
13—20%, []D22 –87.0 (c 1.70, CHCl3) (cf. Ref. 33: []D22 –75.3
(c 0.36, CHCl3)).
Bis(2,3:4,5ꢀdiꢀOꢀisopropylideneꢀ1ꢀdeoxyꢀꢀDꢀfructopyranosꢀ
1ꢀyl) disulfide (16). A dense yellow liquid. The yield was
15—18%, []D22 –55.0 (c 0.67, CHCl3) (cf. Ref. 30: []D22 –54.1
(c 1.00, CHCl3)).
Bis[(1S,2S,5R)ꢀ2ꢀisopropylꢀ5ꢀmethylcyclohexyl] disulfide
(14a). White crystals. M.p. 52—53 C. The yield was 30—35%,
[]D22 +353.0 (c 1.1, Me2CO) (cf. Ref. 34: []D22 +350.0 (c 1.0,
Me2CO)).
Bis[(1S,2S,4S)ꢀ1,7,7ꢀtrimethylbicyclo[2.2.1]heptꢀ2ꢀyl] diꢀ
sulfide (14b). A white powder. M.p. 215—216 C. The yield was
20—22%, []D22 –83.0 (c 0.90, CHCl3) (cf. Ref. 35: []D22 –81.7
(c 0.88, CHCl3)).
22
S, 14.47. C22H34O5S2. Calculated (%): C, 59.70; H, 7.74; S, 14.49.
IR, /cm–1: 519 (S—S), 580 (C—S), 988, 1061 (C—O), 1165
(O—C—O), 1211, 1645 (C=C), 2931. 1H NMR (CDCl3),
: 0.86 (s, 3 H, 9´ꢀMe); 1.16—1.28 (m, 1 H, HA(4´)); 1.31 (s, 3 H,
8´ꢀMe); 1.38 (s, 3 H, 12ꢀMe); 1.47 (s, 3 H, 8ꢀMe); 1.54 (s, 3 H,
11ꢀMe); 1.57 (s, 3 H, 9ꢀMe); 2.06—2.16 (m, 1 H, H(5´)); 2.22
(t, 1 H, H(1´), J = 5.1 Hz); 2.25—2.34 (m, 2 H, H(7´)); 2.42 (dt, 1 H,
HB(4´), J = 8.6 Hz, J = 5.6 Hz); 3.12 (d, 1 H, HA(1), J = 14.0 Hz);
3.35—3.46 (m, 3 H, HB(1), H(10´)); 3.80 (d, 1 H, HA(6),
J = 12.9 Hz); 3.94 (dd, 1 H, HB(6), J = 12.9 Hz, J = 1.4 Hz); 4.26
(d, 1 H, H(5), J = 8.0 Hz); 4.35 (d, 1 H, H(3), J = 2.5 Hz); 4.64 (dd,
1 H, H(4), J = 7.8 Hz, J = 2.6 Hz); 5.50 (s, 1 H, H(3´)). 13C NMR,
: 21.32 (C(9´)), 24.16 (C(12)), 25.59 (C(8)), 25.95 (C(11)),
26.17 (C(8´)), 26.59 (C(9)), 31.43 (C(4´)), 31.73 (C(7´)), 38.16
(C(6´)), 40.48 (C(5´)), 45.35 (C(1´)), 45.85 (C(10´)), 49.53 (C(1)),
61.60 (C(6)), 70.44 (C(4)), 70.82 (C(5)), 72.29 (C(3)), 102.63 (C(2)),
108.59 (C(7)), 109.15 (C(10)), 121.64 (C(3´)), 142.80 (C(2´)).
1,2:3,4ꢀDiꢀOꢀisopropylideneꢀ6ꢀ[(trans)ꢀ2,6,6ꢀtrimethylbiꢀ
cyclo[3.1.1]heptꢀ2ꢀenꢀ4ꢀyldisulfanyl]ꢀ6ꢀdeoxyꢀꢀDꢀgalactopyrꢀ
anose (12e). A dense yellow liquid. The yield was 78% (0.269 g),
Bis[transꢀ(1S,2S,5R)ꢀ6,6ꢀdimethylbicyclo[3.1.1]heptꢀ2ꢀylꢀ
methyl] disulfide (14c). A yellow liquid. The yield was 25—30%,
22
[]D –73.8 (c 0.60, CHCl3). Found (%): C, 70.98; H, 10.17;
S, 18.85. C20H34S2. Calculated (%): C, 70.94; H, 10.12; S, 18.94.
IR, /cm–1: 540 (S—S), 615 (C—S), 874 (C—S), 937, 2910.
1H NMR (CDCl3), : 0.84—1.02 (m, 1 H, HA(7´)); 1.02 (s, 3 H,
8´ꢀMe); 1.23 (s, 3 H, 9´ꢀMe); 1.44—1.66 (m, 1 H, HA(3´));
1.67—2.17 (m, 5 H, H(1´), HB(3´), H(4´), H(5´)); 2.26—2.50
(m, 2 H, H(2´), HB(7´)); 2.70—2.87 (m, 2 H, H(10´)). 13C NMR,
: 21.91 (C(3´)), 23.30 (C(8´)), 26.14 (C(4´)), 27.97 (C(9´)),
33.31 (C(7´)), 38.70 (C(6´)), 40.32 (C(2´)), 41.31 (C(5´)), 45.42
(C(1´)), 46.19 (C(10´)).
22
[]D –84.0 (c 0.91, CHCl3). Found (%): C, 59.57; H, 7.79;
S, 14.42. C22H34O5S2. Calculated (%): C, 59.70; H, 7.74; S, 14.49.
IR, /cm–1: 511 (S—S), 646 (C—S), 1071 (C—O), 1169 (O—C—O),
1
1645 (C=C), 2930. H NMR (CDCl3), : 0.91 (s, 3 H, 9´ꢀMe);
1.37 (s, 6 H, 10´ꢀMe, 12ꢀMe); 1.38 (s, 3 H, 8ꢀMe); 1.38—1.48
(m, 1 H, HA(7´)); 1.37 (s, 3 H, 8ꢀMe); 1.38 (s, 3 H, 12ꢀMe); 1.47
(s, 3 H, 11ꢀMe); 1.61 (s, 3 H, 9ꢀMe); 1.64—1.81 (m, 3 H,
HA(7´)); 1.48 (s, 3 H, 9ꢀMe); 1.62 (s, 3 H, 11ꢀMe); 1.73 (s, 3 H,
8´ꢀMe); 2.06 (t, 1 H, H(5´), J = 5.3 Hz); 2.24—2.34 (m, 1 H,
HB(7´)); 2.40—2.48 (m, 1 H, H(1´)); 2.85—3.02 (m, 2 H, H(6));
3.72—3.81 (m, 1 H, H(4´)); 4.16 (t, 1 H, H(5), J = 6.6 Hz); 4.36
(dd, 1 H, H(2), J = 5.0 Hz, J = 2.2 Hz); 4.37—4.43 (m, 1 H,
H(3)); 4.67 (dd, 1 H, H(4), J = 8.0 Hz, J = 2.2 Hz); 5.27 (s, 1 H,
H(3´)); 5.57 (d, 1 H, H(1), J = 5.0 Hz). 13C NMR, : 20.60
(C(9´)), 22.98 (C(8´)), 24.49 (C(12)), 25.04 (C(8)), 26.00 (C(11)),
26.11 (C(9)), 26.50 (C(10´)), 28.58 (C(7´)), 38.87 (C(6)), 42.72
(C(6´)), 45.26 (C(1´)), 47.70 (C(5´)), 54.41 (C(4´)), 66.58 (C(5)),
70.64 (C(2)), 70.95 (C(4)), 71.58 (C(3)), 96.73 (C(1)), 108.81
(C(7)), 109.29 (C(10)), 115.65 (C(3´)), 149.22 (C(2´)).
Bis[(1S,5R)ꢀ6,6ꢀdimethylbicyclo[3.1.1]heptꢀ2ꢀenꢀ2ꢀylmethꢀ
yl] disulfide (14d). A dense yellow liquid. The yield was 26—30%,
22
[]D –36.8 (c 0.52, CHCl3). Found (%): C, 71.83; H, 9.15;
S, 19.02. C20H30S2. Calculated (%): C, 71.80; H, 9.04; S, 19.17.
IR, /cm–1: 542 (S—S), 599 (C—S), 1645 (C=C), 2918.
1H NMR (CDCl3), : 0.87 (s, 3 H, 9´ꢀMe); 1.17—1.36 (m, 1 H,
HA(4´)); 1.33 (s, 3 H, 8´ꢀMe); 2.07—2.18 (m, 1 H, H(5´)); 2.23
(t, 1 H, H(1´), J = 5.0 Hz); 2.26—2.36 (m, 2 H, H(7´)); 2.44
(td, 1 H, HB(4´), J = 8.6 Hz, J = 5.6 Hz); 3.25—3.39 (m, 2 H,
H(10´)); 5.49 (s, 1 H, H(3´)). 13C NMR, : 21.34 (C(9´)),
26.21 (C(8´)), 31.44 (C(4´)), 31.78 (C(7´)), 38.19 (C(6´)),
40.50 (C(5´)), 45.36 (C(1´)), 45.74 (C(10´)), 121.32 (C(3´)),
143.02 (C(2´)).
2,3;4,5ꢀDiꢀOꢀisopropylideneꢀ1ꢀ[(trans)ꢀ2,6,6ꢀtrimethylbiꢀ
cyclo[3.1.1]heptꢀ2ꢀenꢀ4ꢀyldisulfanyl]ꢀ1ꢀdeoxyꢀꢀDꢀfructopyrꢀ
anose (13e). A dense yellow liquid. The yield was 83% (0.305 g),
Bis[2,6,6ꢀtrimethylbicyclo[3.1.1]heptꢀ2ꢀenꢀ4ꢀyl] disulfide
(14e). A colorless liquid. The yield was 24—26%. Found (%):
C, 71.70; H, 9.14; S, 19.16. C20H30S2. Calculated (%): C, 71.80;
H, 9.04; S, 19.17. IR, /cm–1: 567 (S—S), 611 (C—S), 1645
22
[]D +6.0 (c 1.2, CHCl3). Found (%): C, 59.41; H, 7.64;
1
S, 14.59. C22H34O5S2. Calculated (%): C, 59.70; H, 7.74; S, 14.49.
(C=C), 2924. H NMR (CDCl3), : 0.92 (s, 3 H, 9´ꢀMe); 1.38
IR, /cm–1: 517 (S—S), 573 (C—S), 1067 (C—O), 1165 (O—C—O),
(s, 3 H, 10´ꢀMe); 1.40—1.50 (m, 1 H, HA(6´)); 1.75 (s, 3 H, 8´ꢀMe);
2.06 (t, 1 H, H(5´), J = 5.4 Hz); 2.23—2.35 (m, 1 H, HB(6´));
2.38—2.49 (m, 1 H, H(1´)); 3.68 (s, 1 H, H(4´)); 5.33 (s, 1 H,
H(3´)). 13C NMR, : 20.62 (C(9´)), 22.97 (C(8´)), 26.50 (C(10´)),
28.65 (C(6´)), 42.77 (C(7´)), 45.36 (C(1´)), 47.69 (C(5´)), 55.08
(C(4´)), 116.05 (C(3´)), 148.74 (C(2´)).
Evaluation of toxicity, antioxidant, and membranoprotective
activity of obtained compounds. For in vitro evaluation of toxiꢀ
city, antioxidant and membranoprotective activity, a 0.5% (v/v)
suspension of laboratory mice erythrocytes in the phosphateꢀ
buffered saline (PBS, рH 7.4) was used. Ethanol was used as
a solvent, while acetone was used for symmetric disulfides with
two terpene fragments 14a—e, which are poorly soluble in
ethanol. Diꢀnꢀpropyl disulfide with known antioxidant properꢀ
ties was chosen as a standard.36
1
1645 (C=C), 2932. H NMR (CDCl3), : 0.90 (s, 3 H, 9´ꢀMe);
1.36 (s, 3 H, 10´ꢀMe); 1.37 (s, 3 H, 12ꢀMe); 1.34—1.50 (m, 1 H,
HA(7´)); 1.48 (s, 3 H, 8ꢀMe); 1.52 (s, 3 H, 11ꢀMe); 1.56 (s, 3 H,
9ꢀMe); 1.72 (s, 3 H, 8´ꢀMe); 2.05 (t, 1 H, H(5´), J = 5.4 Hz);
2.28 (dt, 1 H, HB(7´), J = 9.1 Hz, J = 5.6 Hz); 2.41—2.53 (m, 1 H,
H(1´)); 3.07 (d, 1 H, HA(1), J = 13.8 Hz); 3.49 (d, 1 H, HB(1),
J = 13.8 Hz); 3.73—3.98 (m, 3 H, H(4´), H(6)); 4.96 (d, 1 H,
H(5), J = 8.0 Hz); 4.34 (d, 1 H, H(3), J = 2.5 Hz); 4.63 (dd, 1 H,
H(4), J = 7.7 Hz, J = 2.2 Hz); 5.28 (s, 1 H, H(3´)). 13C NMR, :
20.58 (C(9´)), 22.94 (C(8´)), 24.10 (C(12)), 25.60 (C(8)), 25.97
(C(11)), 26.49 (C(9)), 26.60 (C(10´)), 28.55 (C(7´)), 42.68 (C(6´)),
45.21 (C(1´)), 47.71 (C(5´)), 50.75 (C(1)), 55.20 (C(4´)), 61.59
(C(6)), 70.40 (C(4)), 70.80 (C(5)), 72.23 (C(3)), 102.71 (C(2)),
108.57 (C(7)), 109.09 (C(10)), 115.72 (C(3´)), 149.04 (C(2´)).

Hexapyranosyl terpenyl disulfides
Russ.Chem.Bull., Int.Ed., Vol. 64, No. 3, March, 2015
731
Toxicity of compounds was evaluated based on their ability
to induce hemolysis. Solutions of compounds were added to
a suspension of erythrocytes and incubated for 5 h at 37 C in
a Biosan ESꢀ20 temperatureꢀcontrolled shaker (Latvia). Control
samples contained a corresponding solvent (0.1% of a total volꢀ
ume of the incubation mixture). Membranoprotective and antiꢀ
oxidant activity of disulfides were determined based on their
inhibition degree of the induced hemolysis, the inhibition of the
accumulation of secondary products POL and the oxidation of
oxyhemoglobin in erythrocytes. To accomplish this, hemolysis
was initiated with an aqueous solution of hydrogen peroxide
(0.006%) 30 min after the addition of solutions of compounds
under study to the suspension of erythrocytes. Then, the reacꢀ
tion mixture was incubated for 5 h at 37 C in a temperatureꢀ
controlled shaker with a slow stirring.
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An aliquot was collected from the incubation media, centriꢀ
fuged for 5 min (1600 g), hemolysis degree was determined based on
the content hemoglobin in the supernatant on a ThermoSpectroꢀ
mic Genesys 20 spectrophotometer (USA) at  = 524 nm.19
Completely hemolyzed sample served as a reference to calculate
percentage of hemolysis.18 The content of secondary products
POL reacting with 2ꢀthiobarbituric acid was determined spectroꢀ
photometrically.37 To evaluate accumulation of the oxidation
products of hemoglobin, the absorption spectrum of hemolyzate
was analyzed, using a Lyumeks Flyuoratꢀ02ꢀPanorama spectroꢀ
fluorimeter. The contents of oxyhemoglobin (oxyHb), metheꢀ
moglobin (metHb), and ferrylhemoglobin (ferrylHb) were calꢀ
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Oxidative Stress II, Methods in Molecular Biology, Ed. D.
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The authors are grateful to E. N. Zainullina, S. P. Kuzꢀ
netsov, and I. N. Alekseev for their help with recording of
NMR spectra and E. U. Ipatova for recording of IR spectra.
This work was financially supported by the Russian
Foundation for Basic Research (Project No. 13ꢀ03ꢀ01312_a).
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in revised form February 5, 2015