Synthesis and characterization of some novel thiosemicarbazones of substituted benzaldehydes and N-(Hepta-O-Acetyl-β-D-Lactosyl) thiosemicarbazide

Article abstract of DOI:10.1080/07328303.2015.1114119

A set of reaction conditions, including ionic liquids as catalysts, water as solvent and microwave-assisted heating method, had been investigated for the synthesis of hepta-O-acetyl-β-D-lactosyl thiosemicarbazones. Based on optimized conditions, namely, [

Full text of DOI:10.1080/07328303.2015.1114119

Journal of Carbohydrate Chemistry
ISSN: 0732-8303 (Print) 1532-2327 (Online) Journal homepage: http://www.tandfonline.com/loi/lcar20
Synthesis and Characterization of Some
Novel Thiosemicarbazones of Substituted
Benzaldehydes and N-(Hepta-O-Acetyl-β-d-
Lactosyl)Thiosemicarbazide
Nguyen Dinh Thanh, Hoang Thi Kim Van & Truong Thi Thu
To cite this article: Nguyen Dinh Thanh, Hoang Thi Kim Van & Truong Thi Thu (2015) Synthesis
and Characterization of Some Novel Thiosemicarbazones of Substituted Benzaldehydes and
N-(Hepta-O-Acetyl-β-d-Lactosyl)Thiosemicarbazide, Journal of Carbohydrate Chemistry, 34:9,
514-544, DOI: 10.1080/07328303.2015.1114119
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Date: 19 December 2015, At: 20:10

Journal of Carbohydrate Chemistry, 34:514–544, 2015
Copyright ꢀC Taylor & Francis Group, LLC
ISSN: 0732-8303 print / 1532-2327 online
DOI: 10.1080/07328303.2015.1114119
Synthesis and Characterization
of Some Novel
Thiosemicarbazones of
Substituted Benzaldehydes
and N-(Hepta-O-Acetyl-β-D-
Lactosyl)Thiosemicarbazide
1
2
1
Nguyen Dinh Thanh, Hoang Thi Kim Van, and Truong Thi Thu
1Faculty of Chemistry, VNU University of Science, 19 Le Thanh Tong, Hoan Kiem, Ha
Noi, Viet Nam
2
Viet Tri University of Industry, Tien Kien, Lam Thao, Phu Tho, Viet Nam
GRAPHICAL ABSTRACT
A set of reaction conditions, including ionic liquids as catalysts, water as solvent
and microwave-assisted heating method, had been investigated for the synthesis
of hepta-O-acetyl-β-D-lactosyl thiosemicarbazones. Based on optimized conditions,
2 2 3
namely, [HO(CH ) NH ][OAc] as catalyst, water as solvent and 300 W microwave
power, a series of substituted benzaldehyde hepta-O-acetyl-β-D-lactosyl thiosemicar-
bazones was synthesized by reaction of hepta-O-acetyl-β-D-lactosyl thiosemicarbazide
with the corresponding substituted benzaldehydes. The high yields of 90–97% were
achieved. Almost all of the obtained thiosemicarbazones exhibited remarkable an-
tibacterial activity against Bacillus subtilis, Staphylococcus aureus, Staphylococcus
epidermidis, Enterobacter, Escherichia coli, Pseudomonas aeruginosa and Klebsiella
Received August 6, 2015; accepted October 26, 2015.
Address correspondence to Nguyen Dinh Thanh, Faculty of Chemistry, VNU
University of Science, 19 Le Thanh Tong, Hoan Kiem, Ha Noi, Viet Nam. E-mail:
nguyendinhthanh@hus.edu.vn
5
14

N-(Per-O-acetylated β-D-Lactosyl)Thiosemicarbazones
5
15
pneumonia in comparison with sulphamethoxazole and trimethoprim as drug refer-
ences and antifungal activity against Aspergillus niger, Candida albicans and Saccha-
romyces cerevisiae in comparison with clotrimazole as drug reference. MIC values of
the compounds range from 39.1 to 2500 μg/mL for bacterial activity and from 0.25 to
2
5.6 μg/mL for fungal activity.
Keywords Antibacterial; Antifungal; Ionic liquid; β-Lactose; Thiosemicarbazone;
Microwave-assisted reaction
INTRODUCTION
Thiosemicarbazones are compounds that contain azomethine and thiourea
bonds simultaneously. Thiosemicarbazone derivatives of monosaccharide
and disaccharide have received considerable attentions because of their
multifaceted properties. These compounds are known to possess a lot
[
1]
of interesting biological activities, such as antituberculosis,
antimicro-
bial,[
tifungal,[ antitumor,
and enzyme inhibitory activities.
2,3]
antibacterial,
[4,11]
anticancer,
cytotoxic, antioxidant,
[5,6]
anticonvulsant, antidiabetic,[8] an-
anti-dyslipidemic,
In general, thiosemicarbazone derivatives
[7]
9]
[10,11]
[12]
[13–15]
[15]
[
16]
containing monosaccharide moiety have significant anti-microorganisms and
antioxidant activities both in vivo and in vitro.[
13,14,16]
On the other hand, sugar
isothiocyanates are among the most versatile synthetic intermediates in carbo-
[
17,18]
hydrate chemistry.
They can be converted into a broad spectrum of func-
tional groups such as amide, isonitrile, carbodiimide, and also N-thiocarbonyl
derivatives allowing, simultaneously, the covalent coupling of a quite unre-
stricted variety of structures to the saccharide moiety.
In several previous articles, the synthesis of substituted aro-
matic aldehyde/ketone N-(per-O-acetylated glucopyranosyl) were re-
ported.[
17,13,14,20–28]
These compounds were synthesized by reaction of
N-(per-O-acetylglycosyl)thiosemicarbazides with the corresponding car-
bonyl compounds, however, thiosemicarbazones containing simultaneously
disaccharide moiety and benzene ring have not been reported yet.
In recent years, ionic liquids have been used more and more frequently
in chemistry as reaction and/or extraction solvents or as catalysts. Ionic liq-
uids with imidazole ring could solubilize carbonyl compounds, alcohols, alkyl
halides, and also transition metal complexes, depending on the anion and the
[
29–31]
alkyl group of the imidazolium cation.
It’s known that the conventional catalysts for reactions between aldehy-
des or ketones and amines involve mainly organic and mineral acids, such as
[
1,3,6–10,12,13,16]
acetic acid and hydrochloric acid.
The use of these catalysts often
suffers from the drawbacks of prolonged reaction times, harsh and harm-
ful reaction conditions, and sometimes difficulty in product separations. This
problem needs to be addressed in carbohydrate synthesis because of the
susceptibility of the derivatives of carbohydrates towards acids, especially
strong acids. In reactions of peracetylated glucopyranosyl thiosemicarbazides,

16 N. Dinh Thanh et al.
5
this becomes extremely important due to the sensibility of acetate groups to-
ward hydrolysis in the presence of strong acidic and basic catalysts. Therefore,
we are especially interested in developing the use of inexpensive, simple and
efficient catalysts in the synthesis of thiosemicarbazones having monosaccha-
ride moiety. On the other hand, it’s known that Brønsted-acidic task-specific
ionic liquids (TSILs), which are the third generation of ionic liquids, are de-
signed to replace traditional mineral or organic liquid acids as catalysts, such
as sulfuric acid, hydrochloric acid, acetic acid, and p-toluenesulfonic acid in
[
32]
these chemical processes.
Such acidic TSILs exhibit dual role, as solvents
and as catalysts, in different organic reactions.[
33,44–48]
In fact, the use of such
Brønsted-acidic TSILs as catalysts is being developed and explored in many
types of organic reactions. Our report is one of such contributions in develop-
ment and exploration of ionic liquids in general and TSILs in particular.
Continuing previous studies on the synthesis and the reactivity of N-
per-O-acetyl-D-glycopyranosyl)thiosemicarbazides,[
13,28]
in the present paper
(
we report a systematic study on the synthesis, spectral characterization, and
ꢁ
ꢁ
ꢁ
ꢁ
biological activities of a series of substituted benzaldehyde N-(2,3,6,2 ,3 ,4 ,6 -
hepta-O-acetyl-β-D-lactosyl)thiosemicarbazones. The use of acids (acetic and
hydrochloric) and piperidine, as well as ionic liquids, as catalysts was also in-
vestigated in this paper. The reaction was carried out by using conventional
and microwave-assisted heating methods.[ The antibacterial and antifungal
activities of these thiosemicarbazones against some microorganisms were eval-
uated.[
34]
39–42]
RESULTS AND DISCUSSION
Chemistry
ꢁ
ꢁ
ꢁ
ꢁ
Required N-(2,3,6,2 ,3 ,4 ,6 -hepta-O-acetyl-β-lactosyl)thiosemicarbazide 3
was prepared from the corresponding isothiocyanate (Sch. 1) by a proce-
dure similar to that used for other peracetylated glucopyranosyl thiosemi-
[
15,23,24]
carbazides.
It’s known that the conversion of N-(2,3,4,6-tetra-O-acetyl-
β-D-glucopyranosyl)isothiocyanate into the corresponding thiosemicarbazide
could usually be carried out at below room temperature in aprotic solvents,
[
15]
[23,24]
such as dichloromethane
and dioxane,
due to the higher reactivity of
in comparison with aro-
[
13,15,23,24]
peracetylated glucopyranosyl isothiocyanate
[
35]
matic isocyanate
towards water. Previously, we found that the conversion of
peracetylated glycopyranosyl isothiocyanates into the corresponding thiosemi-
carbazides could be performed in a protic solvent, such as absolute ethanol, but
◦
the reaction must be performed at lower temperature (usually 7–10 C) to pre-
vent the transformation of isothiocyanate compounds into thiocarbamic acid
[
8,35]
derivatives.
We realized that the use of absolute ethanol as solvent for this
reaction is of great advantage to work out the reaction due to low solubility of

N-(Per-O-acetylated β-D-Lactosyl)Thiosemicarbazones
5
17
these thiosemicarbazides in ethanol. For improvement of this reaction, we have
[13]
[15]
used an 85% hydrazine solution
instead of 100% hydrazine hydrate.
In
this way, N-(hepta-O-acetyl-β-lactosyl)thiosemicarbazide 3 was obtained from
the corresponding isothiocyanate derivative 2 by reaction with 85% hydrazine
[
15]
hydrate (Sch. 1) using the similar procedure in anhydrous dichloromethane
or in anhydrous dioxane.
[
23,24]
The advantage of using ethanol as a reaction
solvent is that the thiosemicarbazide product is precipitated as soon as it is
formed, hence the isolation of the product became easier, simply by filtering
the product precipitates separated. A high yield of 96% was achieved.
Scheme 1: Synthesis of N-(hepta-O-acetyl-β-D-lactosyl)thiosemicarbazide 3. Conditions: (i)
◦
◦
NH SCN, TBAB, 50 C; 1 h; (ii) 85% hydrazine hydrate, EtOH, 7–10 C.
4
There are several methods for the synthesis of isothiocyanate of sug-
ars.[
23,24,36–38]
In this article, the precursor of 2,3,6,2 ,3 ,4 ,6 -hepta-O-acetyl-β-
ꢁ
ꢁ
ꢁ
ꢁ
ꢁ
ꢁ
ꢁ
ꢁ
lactosyl isothiocyanate 2 was prepared from 2,3,6,2 ,3 ,4 ,6 -hepta-O-acetyl-β-
lactosyl bromide 1 by reaction with lead thiocyanate in toluene[ or xylene.
36]
[37]
In this research, we used the reaction of this bromide derivative with dry
ammonium thiocyanate in anhydrous acetonitrile with tetrabutylammonium
bromide as transfer catalyst for 10 min under microwave-assisted irradiation
[
23,24]
(
modified the Tashpulatov’s method).
This procedure is greener than the
[
25,26]
reported procedures
xylene.
which used harmful lead thiocyanate in toluene or
In the literatures, peracetylated glucopyranosyl thiosemicarbazones, in
particular, and other sugars thiosemicarbazones, in general, were synthesized
under severe conditions: heating with acidic catalysts, such as hydrochlo-
ric and acetic acids, in organic solvents, such as methanol, ethanol, and
propanol.[
1–12,13–27]
The reaction time of these protocols are usually lengthy
(
often from 2 h to 48 h). Therefore, the search for methods with smooth con-
ditions is always laid out. First, we considered the reaction of peracetylated
β-lactosyl thiosemicarbazide 3 with unsubstituted benzaldehyde 4a (R H),
as a model reaction, under the conventional conditions for the synthesis of
thiosemicarbazones (Sch. 2, Table 1, Procedure A). This procedure used ab-
solute ethanol as solvent and glacial acetic acid as catalyst, and the reaction
mixture was heated by the conventional heating method or microwave-assisted
conditions. The product yield was 75% for 2 h under refluxing using conven-
tional heating, while in the case of microwave-assisted heating, the reaction
afforded a yield of 78% in only 5 min.

18 N. Dinh Thanh et al.
5
Scheme 2: Synthesis of benzaldehyde N-(hepta-O-acetyl-β-D-lactosyl)thiosemicarbazones
5
a–s. Conditions: (i) glacial acetic acid, EtOH, MW irradiation, 5 min; (ii) ionic liquid
[
HOCH CH NH ][OAc], water, MW irradiation, 2–3 min.
2 2 3
We have evaluated the effect of microwave power on reaction time and
product yield for these reactions (Table 1). We found that, initially, pulses of
0 s of microwave irradiation at maximum power (800 W) were applied, but the
3
yields were not reproducible, and that it was difficult to maintain the heating
of the reaction mixture. Other high microwave powers from 600 W to 300 W
were also evaluated, and the results were similar, except for at 450 W the
yields were higher (78%). A similarly high yield (78%) was also achieved at
microwave power of 300 W. The microwave power of 300 W was then chosen as
the optimized one and continuous microwave irradiation was evaluated below.
We have also used concentrated hydrochloric acid and piperidine as cat-
alysts in this reaction instead of glacial acetic acid (Table 2). The results ob-
tained in Table 2 showed that the use of hydrochloric acid and piperidine did
not give the desired products 5a, even if the mixture was heated for 10 min
under microwave-assisted conditions. It is possible that HCl had blocked the
free amino group in thiosemicarbazide, therefore its reactivity toward carbonyl
functional group was lost, whereas piperidine could not activate the carbonyl
group in benzaldehyde 4a.
Table 1: Different microwave powers used for synthesis of 5a from 3 and 4a
Entry
Microwave power (Watts)
Yield (%) a
1
2
3
4
5
6
800 b
600 b
60
70
78
78
55
b
450
300 b
b
100
Conventional heating c
75 (for 2 h)
a
Isolated yields.
b
Catalyst: Glacial acetic acid, 1 mmol%; Solvent: absolute ethanol; 5 min MW irradiation.
c
Catalyst: Glacial acetic acid, 1 mmol%; Solvent: absolute ethanol; 2 h refluxing.

N-(Per-O-acetylated β-D-Lactosyl)Thiosemicarbazones
5
19
Table 2: Comparison of ionic liquids used as catalysts for model reaction in
Scheme 1 with 4a (R H) at microwave power of 300 W
5
a: Yield (%) / Time (min) a
Entry
Catalysts
In 96% ethanol
In water
b
1
2
3
4
5
6
7
8
CH
HCl acid
Piperidine b
3
CO
2
H
78 / 5
N.A. / 10
N.A. / 10
50 / 2
88 / 2
96 / 2
18 / 10
N.A. / 10
N.A. / 10
50 / 2
90 / 2
97 / 2
c
b
[Et
[Bmim]HSO
[HO(CH
[HSO
[Hmim]BF
3
NH]HSO
4
b
4
)
2
NH
3
][OAc] b
2
b
3
-Bmim][HSO
4
]
80 / 2
87 / 2
82 / 2
88 / 2
b
4
a
Isolated yields.
b
Molar ratio of the catalyst to benzaldehyde is according to the general procedure (see
Experimental part).
c
for Entry 2: 0.01 mL 36% HCl solution in water and absolute ethanol as solvent.
Continuing our study, we have investigated the catalytic capacity
[51]
[52]
of obtained ionic liquids, such as [Et3NH]HSO4,
[Bmim][HSO4],
and [Hmim][BF4][55] in syn-
thesis of thiosemicarbazones 5a from thiosemicarbazide 3 and benzaldehyde
a (R H), as the optimized microwave power (300 W) was used according
[53]
[54]
[
HO(CH2)2NH3][OAc],
[HSO3-Bmim][HSO4]
4
to Table 2. Because some of the ionic liquids could be miscible with water, we
have used water as solvent in these investigations instead of absolute ethanol.
In addition, we found that the use of absolute ethanol or 96% ethanol did not
change the product yields significantly, with 78% and 77% yields of 5a, respec-
tively. Based on these results, we decided to use 96% ethanol as solvent instead
absolute ethanol in following investigations.
We found that almost all of these ionic liquids increased remarkably the
yields of thiosemicarbazones and decreased the reaction time typically by
2
min. Table 2 indicated that, in almost all cases, the reaction that car-
ried out in water gave the higher yield than the one in 96% ethanol, and
that the product yields went up in the following order of these catalysts:
[
<
Bmim][HSO4] (50%) < [HSO3-Bmim][HSO4] (82%) < [Et3NH][HSO4] (86%)
[Hmim][BF4] (88%) < [HO(CH2)2NH3][OAc] (97%). Clearly, the ionic liquid
HO(CH2)2NH3][OAc] is the most suitable for the model reaction described
[
above. Since then, we found that the ionic liquid [HO(CH2)2NH3][OAc] was
the best catalyst for synthesis of thiosemicarbazones. On the other hand, when
these ionic liquids were used as catalysts, the solvent change from 96% ethanol
to water only alters the product yield inconsiderably. Thus, than advantage of
using ionic liquids as catalysts is that the reactions could be carried out in
water instead of 96% ethanol (Tables 2 and 3), making this process green.

20 N. Dinh Thanh et al.
5
Table 3: Different microwave powers for synthesis of 5a using ionic liquid
[
HO(CH
2
)
2
NH
3
][OAc] as catalyst in water solvent
Entry
Microwave power (Watts)
Yield (%) a,b
1
2
3
4
5
800
600
450
300
100
53
67
97.2
97
60
a
Isolated yields.
b
Reaction time: 2 min. Molar ratio of the catalyst to benzaldehyde is according to the general
procedure (see Experimental part).
Next, we have surveyed the effects of microwave power on the reaction
yield when the ionic liquid [HO(CH2)2NH3][OAc] was used. The results listed
in Table 3 indicated that, similarly to the case of using acetic acid as catalyst in
the ethanol, the product yields were lower when the higher microwave powers
were applied in comparison with in cases of the lower microwave powers. The
yield of products 5a were 97.2% and 97% in microwave powers of 450 W and
300 W, respectively. Therefore, the microwave power of 300 W was both energy-
efficient and eco-friendly (using water as the reaction solvent).
Next, we surveyed further how the amounts of the catalyst might influence
this reaction. When the amount of catalyst was reduced to 5 mol%, we found
that the product performance decreased, although the reaction time was pro-
longed twice. When augmenting the amounts of moles of catalyst from 10 mol%
to 20, 30, 40 and 50 mol%, respectively, the product yield did not change much,
even if the reaction time was maintained or prolonged twice. However, the use
of 40 mol%- and 50 mol%-amounts of this TSIL could raise the yield up to
∼
98%, so in general, the use of a large amount of the catalyst is not beneficial
in this synthetic reaction. As such, the amount of catalyst 10 mol% is the most
appropriate for this reaction (Table 4).
After the optimized conditions for the synthesis of thiosemicarbazone 5a
were discovered, we applied these optimized conditions, i.e., using ionic liquid
[HO(CH2)2NH3][OAc] as catalyst, water as solvent and microwave power of
300 W, to the synthesis of other thiosemicarbazones 5b–t (Table 5). The re-
sults from this table indicated that the obtained product yields by using TSIL
as catalyst were higher than the ones obtained with acetic acid as catalyst
whereas the reaction times were maintained equally or changed a little.
Next, we examined the reusability of the catalyst under microwave-
assisted conditions using the model reaction of benzaldehyde 4a with thiosemi-
carbazide 3 in the presence of 2-hydroxyethylammonium acetate (Table 5, En-
try 1). For this purpose, the amount of reagents were raised up to 50 mmol
and the amount of [HO(CH2)2NH3][OAc] was 50 mmol%. Once the reaction

N-(Per-O-acetylated β-D-Lactosyl)Thiosemicarbazones
5
21
Table 4: Effects of catalytic amounts of [HO(CH
of 5a in water at microwave power of 300 W
2 2 3
) NH ][OAc] on synthetic reaction
a
Yields (%) b
Entry
TSIL, (mmol%)
Time (min)
1
2
3
4
5
6
7
8
9
5
2
4
2
4
2
4
2
4
2
4
2
4
45
47
5
10
10
20
20
30
30
40
40
50
50
97
97.3
97.6
97.8
97.6
97.2
97.1
98
1
1
1
0
1
2
98
98.3
a
TSIL = [HO(CH ) NH ][OAc].
2
2
3
b
Isolated yields. Molar ratio of the catalyst to benzaldehyde is according to the general pro-
cedure (see Experimental part).
Table 5: Comparison of synthesis of benzaldehyde N-(peracetylated
β-lactosyl)thiosemicarbazones 5a–s with acetic acid and [HO(CH ) NH ][OAc] as
2 2 3
catalyst in microwave-assisted conditions
CH
3
CO
2
Ha
TSILb
Entry Compd.
R
Yield (%) c Time (min) Yield (%) c Time (min)
1
2
3
4
5
6
7
8
9
0
1
2
3
4
5
6
7
8
9
5a
5b
5c
5d
5e
5f
5g
5h
5i
H
78
87
83
83
70
76
68
78
78
78
72
76
73
73
78
62
77
77
77
5
5
5
5
5
5
5
5
5
5
5
5
5
5
5
5
5
5
5
97
94
90
90
91
93
97
92
97
97
97
97
92
97
97
97
97
97
96
2
3
3
3
2
2
3
3
2
2
2
2
2
2
2
2
2
2
2
4-NO
3-NO
2-NO
4-F
2
2
2
4-Cl
3-Cl
2-Cl
4-Br
4-Me
4-iPr
1
1
1
1
1
1
1
1
1
1
5j
5k
5l
4-OMe
3-OMe
2-OMe
4-OH
5m
5n
5o
5p
5q
5r
2-OH
3-OMe-4-OH
3-OEt-4-OH
5s
4-NMe
2
a
In 5 mL of 96% ethanol, otherwise, in 5 mL of water.
b
c
TSIL = [HO(CH ) NH ][OAc] in water.
2
2
3
Isolated yields. Molar ratios of the catalyst to benzaldehyde is according to the general
procedure (see Experimental part).

22 N. Dinh Thanh et al.
5
Figure 1: Recycling of [HO(CH ) NH ][OAc] as catalyst for reaction of benzaldehyde 4a
2
2
3
with thiosemicarbazide 3 under microwave-assisted conditions: 3 (50 mmol), 4a (50 mmol)
and [HO(CH ) NH ][OAc] (50 mmol%).
2
2
3
was completed, precipitates was filtered out the water solution and washed
with water to remove ionic liquid out to obtain the crude product. The water-
containing ionic liquid (this TSIL is soluble in water) was evaporated under
reduced pressure and the ionic liquid was recovered and reused. The results of
the first experiment and subsequent experiments were almost all consistent in
yields after five runs without any loss of its activity (97%, 96%, 94%, 92% and
9
0%, Fig. 1).
The structures of the prepared substituted benzaldehyde N-
ꢁ
ꢁ
ꢁ
ꢁ
(
2,3,6,2 ,3 ,4 ,6 -hepta-O-acetyl-β-D-lactosyl)thiosemicarbazones 5a–s were
confirmed by their spectroscopic data. The IR spectra show characteristic
−
1
absorptions in the range of 3540–3480 (νOH) and 3348–3159 cm
(νNH),
ester),
−1
1
737–1753 (ν
ester), 1216–1252 and 1044–1056 cm
(ν
C=O
COC
1
−
1
−1
1367–1377 cm
(νC S), and 1580–1615 cm
(νCH N). In the H NMR
spectra, signal of proton NH-2 was found farther downfield in region of δ
1.74–10.69 ppm (in singlet) as this proton was deshielded by anisotropic
1
effect of adjacent imine bond, whereas the signal in region of δ 8.72–8.44 ppm
belongs to NH-4, which was split into a doublet by one neighboring proton on
ꢁ
the carbon C-1 of lactose moiety. The coupling constant in this case was J =
8
.5–9.5 Hz. The protons of the disaccharide moiety had the signals in region
ꢁ
ꢁ
ꢁꢁ
of δ = 5.90–3.80 ppm. The protons in the carbons C-1 and C-2 magnetically
interacted with each other with the coupling constant of J = 7.5–8.0 Hz. This
corresponded with β-(1→4)-glycoside linkage between ring A and ring B in
β-D-lactose (see Sch. 1). In addition, the spin-spin splitting pattern between
H-3 and H-4 on ring B (D-galactose component) differed from the one
between H-3 and H-4 on ring A (D-glucose component). The values of coupling
ꢁ
ꢁ
ꢁꢁ
ꢁ
ꢁ
constants were J = 2.5–3.75 Hz (in ring B) and J = 9.25–9.75 Hz (in ring A),

N-(Per-O-acetylated β-D-Lactosyl)Thiosemicarbazones
5
23
ꢁ
respectively. The chemical shift of proton H-1 (on ring A) was in region of δ =
5
.90–5.74 ppm as triplet pattern. This proton also had similar interaction with
ꢁ
proton H-2 with the coupling constant of J = 9.00–9.25 Hz. The value of this
coupling constant was consistent with a 1,2-trans relationship between these
protons and showed that synthesized hepta-O-acetyl-β-D-lactosyl thiosemicar-
bazones had β-D-anomeric configuration. Proton in azomethine group showed
a sharp peak at δ 7.97–8.18 ppm (CH
N) in singlet. The positions of reso-
nance signals of protons NH-2, NH-4 and CH N were affected significantly
by the substituent nature on the benzene ring: the e-donating groups, such as
4
-NMe2, 4-OH, 4-OCH3 and 4-CH3, caused signal of proton NH-2 shifted to
upfield region, and the e-withdrawing ones, such as 4-NO2, 3-NO2, 4-F, 4-Br,
-Cl and 3-OCH3 and 3-OCH3-4-OH, caused this signal shifted to downfield
region.
The 13C-NMR spectrum of compound 5a–s showed the four-parted regions
of signals at δ 179.7–169.1, 159.7–111.3, 82.1–61.5 and 21.7–20.2 ppm. Sig-
nals at δ 179.1–177.8 ppm belonged to the carbon atom in the C S group.
4
The acetyl groups in these compounds showed resonance at δ 171.7–169.1 ppm
for carbonyl carbon atom and 21.7–20.2 ppm for the methyl carbon one. The
presence of imine in molecule were confirmed by signal at δ 147.8–140.2 ppm,
belong to carbon atom in the azomethine group. It was observed that the NMR
signal of carbon atom in the imine group was affected by these substituents
in the following trend: the e-donating groups, such as 4-NMe2, 4-OH, 4-OCH3
and 4-CH3, caused signal shifted to upfield region, and the e-withdrawing ones,
such as 4-NO2, 3-NO2, 4-F, 4-Br, 4-Cl and 3-OCH3 and 3-OCH3-4-OH, caused
the carbon signal shifted to downfield region.
Biological Evaluation
Antibacterial Activity
Antibacterial activity studies on thiosemicarbazones 5a–s showed that
these compounds had mild to moderate antibacterial activity against the tested
bacteria (Table 6). Moreover, all compounds are more active (lower MIC) than
the references sulphamethoxazole and trimethoprim for the majority of tested
bacteria. It’s known that sulfamethoxazole (SMZ or SMX) is a sulfonamide
[
43]
bacteriostatic antibiotic
and trimethoprim (TMP)-an antibiotic used mainly
in the treatment of urinary tract infections.[ These drugs are most often
used as part of a synergistic combination with trimethoprim in a 5:1 ratio
in co-trimoxazole, and known under trade names such as Bactrim, Septrin,
44]
[
45]
or Septra, Biseptol . . .
Its primary activity is against susceptible forms of
Streptococcus, Staphylococcus aureus (including MRSA-Methicillin-resistant
Staphylococcus aureus), Escherichia coli, Enterobacter spp., Klebsiella pneu-
moniae, Mycobacterium tuberculosis, Salmonella typhi (typhoid fever) and oral

24 N. Dinh Thanh et al.
5
Table 6: Antibacterial activities of thiosemicarbazones 5a–s
Microorganisms/ MIC (μg/mL)
Gram positive
Gram negative
Compound/Drug B.s.a S.a.a S.e.a En.a E.c.a P. a.a
K.p.a logP (calc.)
5
5
5
5
5
5
5
5
5
5
5
5
5
5
5
5
5
5
5
a
b
c
d
e
f
g
h
i
1200 225.1 2500
1200 112.7 1200 225.4 225.4 185.7 175.3
1200 215.3 1200 435 225.4 185.7 156.4
1200 225.5 1200 225.4 225.4 185.7 183.7
1200 58.3 1200 215.5 47.5 185.7 172.1
63.5 58.3 175.3
54.5 185.7 39.1
435
435 225.4 185.1
4.32
4.24
4.23
4.42
4.53
5.08
5.15
5.01
5.25
4.78
5.66
4.45
4.41
4.05
4.07
4.29
3.95
4.48
4.49
0.89
0.79
1200 185.7 1200
435
435 185.4
435 187.1
1200
1200
435
433
85.7 185.7 185.7
435 235.4 435 275.2
87.7
58.3
55.3
58.8
58.3
39.1
39.1
58.7
39.1
j
k
l
225.4 245.8 435
435
1200 255.1 435 245.9 185.2 165.7
1200 175.7 1200
1200 215.3 435 265.4 187.4 177.1
1200 165.7 1200 435 175.8
1200 245.4 435 215.3 165.1 155.3
1200 181.7 1200 435 155.2 58.3 195.1
1200 228.4 435 255.3 190.3 185.1 39.1
1200 176.7 1200 435 174.7 58.3 188.4
1200 235.4 435 215.6 185.7 155.5 39.1
435 182.0
58.3
m
n
o
p
q
r
59.1 175.9
39.1
s
b
c
Ref. A
Ref. B
3200
19.5 3000 3000 3200 3000 156.2
3000 2500 58.32 2500 2500 3000
1100
a
B.s.: Bacillus subtilis; S.a.: Staphylococcus aureus; S.e.: Staphylococcus epidermidis;
En.:Enterobacter; E.c.: Escherchia coli; P. a.: Pseudomonas aeruginosa; K.p.: Klebsiella pneu-
monia
b
Ref. A: Sulphamethoxazole
c
Ref. B: Trimethoprim
[
45,46]
anaerobes.
fections.
pensive.
Sulfamethoxazole is commonly used to treat urinary tract in-
This drug is available as a generic medication and is not very ex-
[
46]
The corresponding MIC values of these references against tested bacteria
were 3200, 19.5, 3000, 3000, 3200, 3000 and 156.2 μg/mL (for sulphamethox-
azole) and 3000, 2500, 58.32, 2500, 2500, 3000 and 1100 μg/mL (for trimetho-
prim), respectively. The data in Table 6 also showed that thiosemicarbazones
5a–s were more active references for almost all tested bacteria, B. subtilis
MIC = 225.4–1200 μg/mL, S. aureus MIC = 176.7–435 μg/mL, S. epidermidis
MIC = 435–2500 μg/mL, Enterobacter MIC = 185.4–435 μg/mL, E. coli MIC
=
174.7–225.4 μg/mL, P. aeruginosa MIC = 58.3–225.4 μg/mL, K. pneumonia
MIC = 39.1–195.1 μg/mL, except S. aureus and K. pneumonia: MIC values
of sulfamethoxazole for this bacterium were 19.5 μg/mL and 156.2 μg/mL,
respectively. For trimethoprim, almost all tested bacteria were inhibited by
these compounds more than reference, except S. epidermidis: MIC value in

N-(Per-O-acetylated β-D-Lactosyl)Thiosemicarbazones
5
25
Table 7: Antifungal activities of thiosemicarbazones 5a–s
MIC (μg/mL)
Compound/Drug A. niger C. albicans F. oxysporum S. cerevisiae logP (calc.)
5
5
5
5
5
5
5
5
5
5
5
5
5
5
5
5
5
5
5
a
b
c
d
e
f
g
h
i
25.6
19.5
7.7
0.50
0.53
0.54
0.51
0.28
0.45
0.34
0.25
0.25
0.57
0.58
0.57
0.25
0.57
0.25
0.25
0.57
0.58
0.57
0.32
4.9
2.7
2.7
4.5
2.7
1.2
2.7
4.4
2.2
2.7
9.4
4.7
2.2
2.7
2.2
2.2
2.7
9.1
4.7
4.9
3.8
3.8
2.6
2.2
2.3
2.2
2.3
2.3
2.3
3.5
3.6
3.2
2.3
3.5
2.3
2.3
3.5
3.6
3.2
2.6
4.32
4.24
4.23
4.42
4.53
5.08
5.15
5.01
5.25
4.78
5.66
4.45
4.41
4.05
4.07
4.29
3.95
4.48
4.49
5.44
25.6
25.6
7.8
19.5
7.7
7.7
j
k
l
7.7
7.7
7.3
m
n
o
p
q
r
7.7
7.7
7.7
7.7
7.7
7.7
s
7.3
Clotrimazole
2.6
this case was 58.32 μg/mL, whereas MIC values of thiosemicarbazones were
in 425–2500 μg/mL. Especially, the most active thiosemicarbazones against
each bacterium were as follows: 5i and 5j against B. subtilis (MIC = 435
and 225.4 μg/mL, respectively), 5e against S. aureus (MIC = 58.3 μg/mL),
5
g–k,m,o,q,s against S. epidermidis (MIC = 435 μg/mL each) and 5g,i–m,q
against K. pneumoniae (MIC = 39.1–58.7 μg/mL), 5e–j against E. coli (MIC =
μg/mL), 5f,i,j against P. aeruginosa (MIC = 55.3–59.1 μg/mL), and 5b,d,e,g-
i,k,m,o,q,s against Enterobacter (MIC = 185.4–275.2 μg/mL).The increased
activity of the synthesized compound comparing to the references, trimetho-
prim and sulphamethoxazole, could be explained by electron delocalization
over the whole molecule. The other reason could be the increased lipophilic
character of thiosemicarbazones 5a–s with logP in the range of 3.95 to 5.66,
as compared to that of the references trimethoprim (log P = 0.79) and sul-
phamethoxazole (log P = 0.89), thus favoring the permeation of 5a–s through
the lipid layer of the bacterial membranes. It may be suggested that this
molecule deactivate various cellular enzymes, which play a vital role in var-
ious metabolic pathways of these microorganisms.[
47]
Antifungal Activity
The biological data in Table 7 showed that, notably, almost all tested
thiosemicarbazones 5a–s had more remarkable activities against these fungi

26 N. Dinh Thanh et al.
5
C. albicans, F. oxysporum, and S. cerevisiae but were more resistant to A.
niger than clotrimazole (the reference compound, MIC = 2.6, 0.32, 4.9 and
2.6 μg/mL, respectively). Clotrimazole is an antifungal medication, commonly,
used in the treatment of fungal infections, in both humans and other animals,
such as vaginal yeast infections (candidiasis), oral thrush (oral candidiasis),
[
48]
and ringworm (dermatophytosis).
This drug is on the WHO’s list of Essen-
tial Medicines (the most important medication needed in a basic health sys-
tem).[
49]
In case of fungus Aspergillus niger, all compounds were less active than
clotrimazole (MIC = 2.6 μg/mL), compounds 5a,b,d,e,g,m had the worst
ability to inhibit to this fungus (MIC = 13.2–25.1 μg/mL), and the re-
maining compounds had medium activity (MIC = 7.3–7.8 μg/mL) but were
less active than clotrimazole. In case of fungus Candida albicans, almost
all compounds were less active than clotrimazole (MIC = 0.42–0.58 and
0
5
.32 μg/mL, respectively), except 5g is equipotent (MIC = 0.34 μg/mL),
e,h,i,m,o,p are more active (MIC = 0.25–0.28 μg/mL). Almost all thiosemi-
carbazones had activities similarly with clotrimazole against fungus Fusar-
ium oxysporum (MIC = 2.4 g/mL), in that, the compounds 5b,c,e,f,g,i,j,m,o,p
have MIC = 2.2–2.4 μg/mL, except thiosemicarbazone 5f is the most active
(
9
MIC = 1.2 g/mL) and thiosemicarbazones 5k, 5r had lowest activity (MIC =
.1 μg/mL). Thiosemicarbazones 5a,b,j,k,l,n were less active against Saccha-
romyces cerevisiae (MIC = 2.8–3.6 μg/mL), and the remaining compounds were
more active (MIC = 2.2–2.4 μg/mL) than clotrimazole (MIC = 2.6 μg/mL).
CONCLUSION
In conclusion, we evaluated in this research the catalysis capacities of selected
ionic liquids in synthetic reactions of thiosemicarbazones having β-lactose moi-
ety under microwave-assisted conditions. We have given an interestingly sim-
ple procedure, cleaner reactions, and use of inexpensive and reusable ionic liq-
uids as catalysts in water, allowing for economic and environmental-friendly
conditions. Almost all quantitative yields and simple isolation and purification
procedures of the products make it a useful procedure for the synthesis of these
compounds. Almost all obtained thiosemicarbazones exhibited the remarkable
antibacterial activity against B. subtilis, S. aureus, S. epidermidis, Enterobac-
ter, E. coli, P. aeruginosa and K. pneumonia and antifungal activity against A.
niger, C. albicans and S. cerevisiae.
EXPERIMENTAL
All solvents, chemicals, and reagents were obtained commercially and used
without purification. Melting points were determined by open capillary method

N-(Per-O-acetylated β-D-Lactosyl)Thiosemicarbazones
5
27
on STUART SMP3 instrument (BIBBY STERILIN, UK) and are uncorrected.
FTIR spectra (KBr disc) were recorded on an Impact 410 FT-IR Spectrometer
1
13
(
Nicolet, USA). H and C NMR spectra were recorded on Bruker Avance Spec-
trometer AV500 (Bruker, Germany) at 500 MHz and 125 MHz, respectively,
using DMSO-d6 as solvent and TMS as an internal standard. Mass spectra
were recorded on mass spectrometer LC-MS LTQ Orbitrap XL (ThermoScien-
tific, USA) in methanol using ESI method or AutoSpec Premier (Water, USA)
using EI method. The entire microwave heating experiments were conducted
under reaction conditions of power and temperature. Thin-layer chromatogra-
phy was performed on silica gel pates 60F254 No. 5715 (Merck, Germany) with
◦
ethyl acetate and light petroleum (bp 60–90 C) or toluene, and spots were visu-
alized with UV light or iodine vapor. The synthesis of some task-specific ionic
liquid was carried out by modifying procedure from a similar method in the
literature, as described as below. The synthesis of [Bmim]BF4 was reported
[
50]
[51]
[52]
previously.
[
Other ionic liquids such as [Et3NH]HSO4,
[Bmim][HSO4],
HO(CH2)2NH3][OAc],[53] [HSO3-Bmim][HSO4][54] and [Hmim][BF4][55] were
prepared by the procedure modified the ones in the corresponding references
as indicated. The ionic liquid was formed quantitatively and in high purity.
Hepta-O-acetyl-β-D-lactosyl bromide 1 was prepared from β-lactose by the sim-
ilar procedure by previously Lemieux’s method.[
56]
ꢀ
ꢀ
ꢀ
ꢀ
Synthesis of 2,3,6,2 ,3 ,4 ,6 -hepta-O-acetyl-β-D-lactosyl
isothiocyanate (2)
To the mixture of hepta-O-acetyl-β-D-lactosyl bromide 1 (7.0 g, 10 mmol)
and dry ammonium thiocyanate (1.9 g, 25 mmol) in 50 mL of absolute acetoni-
trile was added tetrabutylammonium bromide (10 mmol%) under stirring. The
mixture was then irradiated for 10 min at 600 W. Upon completion (monitored
by TLC, the eluent solvent system was toluene/ethyl acetate 1:1 v/v), the pre-
cipitate was removed by filtration and the solvent was removed by distillation,
the residue was dissolved in chloroform, and the solution was washed with
water. The chloroform layer was dried over CaCl2 and evaporated to dryness
in vacuo, and the residue was crystallized from ether-hexane (2:1) to afford
◦
[57]
◦
isothiocyanate 2 (6.09 g). Yield: 90%. M.p. 163–164 C; ref. : 163–165 C.
ꢀ
ꢀ
ꢀ
ꢀ
Synthesis of N-(2,3,6,2 ,3 ,4 ,6 -hepta-O-acetyl-β-D-lactosyl)
thiosemicarbazide (3)
To a solution of hepta-O-acetyl-β-D-lactosyl isothiocyanate 2 (6.77 g,
10 mmol) in 40 mL of absolute ethanol a solution of 85% hydrazine hydrate
(
10 mmol, 1.2 ml) in 10 mL of absolute ethanol was added dropwise slowly
◦
with stirring in 30 minutes so that the reaction temperature is below 10 C.
The white precipitate appeared immediately when several drops of hydrazine

28 N. Dinh Thanh et al.
5
had added due to low solubility of this thiosemicarbazide in ethanol. The tem-
◦
perature of solution was maintained between 10−12 C. The mixture was con-
◦
tinued stirring at 20 C for 30 min. The solid product then was isolated by
filtering with suction. The crude product was crystallized from 96% ethanol to
◦
afford thiosemicarbazide 3 (6.81 g). Yield 96%, mp 187−188 C; FTIR (KBr): ν
–
1
1
(
cm ) 3473, 3409, 3273, 1753, 1620, 1535, 1378, 1226, 1050; H NMR (DMSO-
ꢁ
d6): δ (ppm) 9.21 (s, 1H, NH-2), 8.09 (s, 1H, NH-4), 5.78 (s br, 1H, H-1 ), 5.21 (d,
ꢁ
ꢁ
ꢁ
1
H, J = 2.0 Hz, H-4 ), 5.20 (s br, 1H, H-3 ), 5.12 (dd, 1H, J = 10.25, 3.75 Hz,
ꢁ
ꢁ
ꢁ
ꢁꢁ
H-3 ), 4.95 (t, 1H, J = 9.25 Hz, H-2 ), 4.84 (t, 1H, J = 9.25 Hz, H-2 ), 4.75
ꢁ
ꢁ
(
d, 1H, J = 8.0 Hz, H-1 ), 4.56 (s br, 2H, NH-1), 4.26 (d, 1H, J = 12.0 Hz,
ꢁ
ꢁ
ꢁ
ꢁꢁ
H-5 ), 4.21 (t, 1H, J = 6.25 Hz, H-6 a), 4.01–4.00 (m, 3H, H-6 b, H-6 a, H-
ꢁ
ꢁ
ꢁ
ꢁ
13
6
(
8
5
b), 3.77–3.76 (m, 2H, H-4 , H-5 ), 2.09–1.93 (7s, 21H, 7×CH3CO); C NMR
ꢁ
ꢁ
DMSO-d6): δ (ppm) 182.1 (C
S), 170.2–169.1 (7C, 7×COCH3), 99.7 (C-1 ),
ꢁ
ꢁ
ꢁ
ꢁ
ꢁ
ꢁꢁ
0.8 (C-1 ), 76.1 (C-4 ), 73.3 (C-5 ), 72.6 (C-3 ), 70.8 (C-2 ), 70.4 (C-3 ), 69.7 (C-
ꢁ
ꢁ
ꢁꢁ
ꢁꢁ
ꢁ
ꢁꢁ
), 68.8 (C-2 ), 67.1 (C-4 ), 62.3 (C-6 ), 60.9 (C-6 ), 20.6–20.3 (7C, 7×COCH3);
+
+
ESI-MS: 732.16 [M+Na] , 710.18 [M] , 331.31, 317.27; calc. for C27H39N3O17S
=
709.20 Da; [M+Na] = 732.19 Da; Anal. calcd. for C27H39N3O17S: C,
4
5.70; H, 5.54; N, 5.92; S, 4.52. Found: C, 45.74; H, 5.51; N, 5.95; S,
.50.
4
ꢀ
ꢀ
ꢀ
ꢀ
General procedure for synthesis of N-(2,3,6,2 ,3 ,4 ,6 -hepta-O-
acetyl-β-D-lactosyl)thiosemicarbazones (5a-s)
Procedure A (under refluxing microwave-assisted conditions)
ꢁ
ꢁ
ꢁ
ꢁ
A
mixture of N-(2,3,6,2 ,3 ,4 ,6 -hepta-O-acetyl-β-D-lactosyl)thiosemi
carbazide 3 (1 mmol) and appropriate substituted benzaldehydes 4a-s
1 mmol) in 5 mL of absolute ethanol was added the appropriate acid or
(
base catalyst (1 mmol%, Tables 2, Entry 1–3, for evaluation of the corre-
sponding catalysts) under stirring. The mixture was then irradiated for
a certain time (Table 5, for synthesis of thiosemicarbazones 5a–s). Upon
completion (monitored by TLC, the eluent solvent system was toluene/ethyl
acetate 1:1 v/v), the solvent was evaporated to one half the original vol-
umes. The resulting colorless crystals were filtered by suction. The crude
product when recrystallized from 96% ethanol to afford the title compounds
5a–s.
Procedure B (under microwave-assisted conditions and TSIL as catalyst)
ꢁ
ꢁ
ꢁ
ꢁ
A
mixture of N-(2,3,6,2 ,3 ,4 ,6 -hepta-O-acetyl-β-D-lactosyl)thiosemi
carbazide 3 (1 mmol) and appropriate substituted benzaldehydes 4a-s
1 mmol) in 5 mL of 96% ethanol was added the appropriate ionic liquid cata-
(
lyst (1 mmol%, Tables 2 and 5, Entry 4–8, for evaluation of the corresponding
catalysts) under stirring. The mixture was then irradiated for a certain

N-(Per-O-acetylated β-D-Lactosyl)Thiosemicarbazones
5
29
time (Table 5, for synthesis of thiosemicarbazones 5a–s). Upon completion
(monitored by TLC, the eluent solvent system was toluene/ethyl acetate 1:1,
v/v), the precipitated crude product was collected by filtration, washed by
water and recrystallized from 96% ethanol to afford pure thiosemicarbazones
5
a–s. After removing off the water under reduced pressure to 5 mL volume,
the filtrate containing the ionic liquid could be reused directly in the next run
without further purification.
The products were confirmed by FTIR, H NMR, 13C NMR, MS and physi-
1
2
5
cal data (m.p., [α] ).
D
ꢁ
ꢁ
ꢁ
ꢁ
Benzaldehyde N-(2,3,6,2 ,3 ,4 ,6 -hepta-O-acetyl-β-D-lactosyl)
thiosemicarbazone (5a)
From 709 mg of 3 and 106 mg of 4a. Recrystallized from 96% ethanol to
yield white crystals of 5a. Yield: 622 mg, 78% (reaction time: 5 min, procedure
◦
25
A); 773 mg, 97% (reaction time: 2 min, procedure B); mp 158–159 C; [α] +85.4
D
–
1
(
c 1.0, CHCl3); FTIR (KBr) ν / cm : 3469, 3304, 1750, 1605, 1369, 1222, 1051;
1
H NMR (DMSO-d6): δ (ppm) 11.95 (s, 1H, NH-2), 8.67 (d, 1H, J = 9.5 Hz, NH-
ꢁ
ꢁꢁ
ꢁꢁꢁ
4
), 8.10 (s, 1H, CH N), 7.82–7.80 (m, 2H, H-2 & H-6 ), 7.45–7.43 (m, 3H,
ꢁ
ꢁꢁ
ꢁꢁꢁ
ꢁꢁꢁ
ꢁ
H-3 , H-4 & H-5 ), 5.86 (t, 1H, J = 9.25 Hz, H-1 ), 5.31 (t, 1H, J = 9.25 Hz,
ꢁ
ꢁꢁ
ꢁ
H-3 ), 5.24 (d, 1H, J = 3.5 Hz, H-4 ), 5.19 (t, 1H, J = 9.5 Hz, H-2 ), 5.15 (dd,
ꢁ
ꢁ
ꢁꢁ
1
H, J = 10.5, 3.5 Hz, H-3 ), 4.87 (dd, 1H, J = 10.0, 3.0 Hz, H-2 ), 4.80 (d, 1H,
ꢁ
ꢁ
ꢁ
J = 8.0 Hz, H-1 ), 4.30 (d, 1H, J = 11.0 Hz, H-6 a), 4.26 (t, 1H, J = 6.75 Hz,
ꢁ
ꢁ
ꢁ
H-5 ), 4.07 (dd, 1H, J = 12.0, 5.5 Hz, H-6 b), 4.04 (dd, 1H, J = 6.25, 2.25 Hz,
ꢁ
ꢁ
ꢁꢁ
H-6 a), 4.03 (dd, 1H, J = 6.25, 2.25 Hz, H-6 b), 3.89 (ddd, 1H, J = 9.75, 5.75,
ꢁ
ꢁ
1
.75 Hz, H-5 ), 3.80 (t, 1H, J = 9.5 Hz, H-4 ), 2.11–1.90 (s, 21H, 7 × COCH3);
1
3
C NMR (DMSO-d6): δ (ppm) 178.3 (C S), 170.3–169.1 (7 × COCH3), 143.7
ꢁ
ꢁꢁ
ꢁꢁꢁ
ꢁꢁꢁ
ꢁꢁꢁ
ꢁꢁꢁ
ꢁ
(
5
(
(
CH N), 133.7 (C-1 ), 130.3 (C-4 ), 128.7 (C-2 & C-6 ), 127.6 (C-3 & C-
ꢁ
ꢁꢁ
ꢁꢁ
ꢁ
ꢁ
ꢁ
ꢁ
), 99.6 (C-1 ), 81.2 (C-1 ), 76.1 (C-4 ), 73.3 (C-5 ), 72.7 (C-3 ), 71.1 (C-2 ), 70.4
ꢁ
ꢁ
ꢁꢁ
ꢁꢁ
ꢁꢁ
ꢁ
ꢁꢁ
C-3 ), 69.7 (C-5 ), 68.8 (C-2 ), 67.1 (C-4 ), 62.4 (C-6 ), 61.0 (C-6 ), 20.7–20.3
+
+
7×COCH3); ESI-MS: m/z: 820.34 [M+Na] , 798.39 [M+H] , 766.59, 738.50,
58.66, 541.45, 331.48; calc. for C34H43N3O17S = 797.23 Da. Anal. calcd. for
C34H43N3O17S: C, 51.19; H, 5.43; N, 5.27; S, 4.02. Found: C, 51.23; H, 5.41; N,
6
5.25; S, 4.05.
4
-Nitrobenzaldehyde
ꢁ
ꢁ
ꢁ
ꢁ
N-(2,3,6,2 ,3 ,4 ,6 -hepta-O-acetyl-β-D-lactosyl)thiosemicarbazone (5b)
From 709 mg of 3 and 151 mg of 4b. Recrystallized from 96% ethanol to
yield white crystals of 5b. Yield: 732 mg, 87% (reaction time: 5 min, proce-
◦
25
D
dure A); 791 mg, 94% (reaction time: 3 min, procedure B); mp 172–173 C; [α]
–
1
+
79.8 (c 1.0, CHCl3); FTIR (KBr) ν / cm : 3490, 3325, 1746, 1615, 1376, 1231,
1
1
056; H NMR (DMSO-d6): δ (ppm) 12.16 (s, 1H, NH-2), 8.88 (d, 1H, J = 9.0 Hz,
ꢁ
ꢁꢁ
ꢁꢁꢁ
NH-4), 8.25 (d, 2H, J = 9.0 Hz, H-3 & H-5 ), 8.18 (s, 1H, CH N), 8.10 (d,
ꢁꢁ
ꢁꢁꢁ
ꢁ
2
H, J = 9.0 Hz, H-2ꢁ & H-6 ), 5.88 (t, 1H, J = 9.0 Hz, H-1 ), 5.31 (t, 1H,

30 N. Dinh Thanh et al.
5
ꢁ
ꢁ
ꢁꢁ
J = 9.25 Hz, H-3 ), 5.24–5.21 (m, 2H, H-2 & H-4 ), 5.15 (dd, 1H, J = 10.0,
ꢁ
ꢁ
ꢁꢁ
ꢁꢁ
3
.5 Hz, H-3 ), 4.88 (t, 1H, J = 8.75 Hz, H-2 ), 4.80 (d, 1H, J = 8.0 Hz, H-1 ),
ꢁ
ꢁꢁ
4
.31 (d, 1H, J = 11.5 Hz, H-6 a), 4.25 (t, 1H, J = 6.5 Hz, H-5 ), 4.07 (dd, 1H,
ꢁ
ꢁꢁ
ꢁꢁ
J = 12.5, 5.5 Hz, H-6 b), 4.03 (d, 2H, J = 6.0 Hz, H-6 a & H-6 b), 3.90–3.87
ꢁ
ꢁ
(
m, 1H, H-5 ), 3.81 (t, 1H, J = 9.25 Hz, H-4 ), 2.11–2.01 (s, 21H, 7×COCH3);
3
1
C NMR (DMSO-d6): δ (ppm) 178.8 (C
S), 170.2–169.0 (7×COCH3), 147.8
ꢁ
ꢁꢁ
ꢁꢁꢁ
ꢁꢁꢁ
ꢁꢁꢁ
ꢁꢁꢁ
ꢁ
(
5
(
(
C-4 ), 141.1 (CH N), 140.2 (C-1 ), 128.4 (C-2 & C-6 ), 123.8 (C-3 & C-
), 99.6 (C-1 ), 81.3 (C-1 ), 76.0 (C-4 ), 73.4 (C-5 ), 72.8 (C-3 ), 71.2 (C-2 ), 70.4
C-3 ), 69.7 (C-5 ), 68.9 (C-2 ), 67.1 (C-4 ), 62.4 (C-6 ), 61.0 (C-6 ), 20.7–20.3
ꢁ
ꢁꢁ
ꢁꢁ
ꢁ
ꢁ
ꢁ
ꢁ
ꢁ
ꢁ
ꢁꢁ
ꢁꢁ
ꢁꢁ
ꢁ
ꢁꢁ
+
7×COCH3); ESI-MS: m/z: 843.80 [M+H] , 840.91, 799.01, 668.15, 443.28,
3
66.45; calc. for C34H42N4O19S = 842.22 Da. Anal. calcd. for C34H42N4O19S:
C, 48.45; H, 5.02; N, 6.65; S, 3.80. Found: C, 48.42; H, 5.06; N, 6.68; S,
3.83.
3
-Nitrobenzaldehyde
ꢁ
ꢁ
ꢁ
ꢁ
N-(2,3,6,2 ,3 ,4 ,6 -hepta-O-acetyl-β-D-lactosyl)thiosemicarbazone (5c)
From 709 mg of 3 and 151 mg of 4c. Recrystallized from 96% ethanol to
yield white crystals of 5c. Yield: 699 mg, 83% (reaction time: 5 min, procedure
◦
25
A); 758 mg, 90% (reaction time: 3 min, procedure B); mp 173–174 C; [α] +94.7
D
–
1
(
c 1.0, CHCl3); FTIR (KBr) ν / cm : 3548, 3340, 1751, 1615, 1372, 1229, 1048;
1
H NMR (DMSO-d6): δ (ppm) 12.10 (s, 1H, NH-2), 8.83 (d, 1H, J = 9.0 Hz, NH-
ꢁ
ꢁꢁ
ꢁꢁꢁ
4
(
9
(
), 8.21 (s, 1H, CH N), 8.58 (s, 1H, H-2 ), 8.30 (d, 1H, J = 7.5 Hz, H-4 ), 7.72
ꢁ
ꢁꢁ
ꢁꢁꢁ
d, 1H, J = 8.0 Hz, H-5 ), 8.24 (dd, 1H, J = 8.0, 1.5 Hz, H-6 ), 5.86 (t, 1H, J =
ꢁ
ꢁ
ꢁꢁ
.0 Hz, H-1 ), 5.31 (t, 1H, J = 9.0 Hz, H-3 ), 5.24 (d, 1H, J = 3.5 Hz, H-4 ), 5.20
ꢁ
ꢁꢁ
t, 1H, J = 9.5 Hz, H-2 ), 5.15 (dd, 1H, J = 10.0, 3.5 Hz, H-3 ), 4.88 (dd, 1H, J =
ꢁ
ꢁ
ꢁꢁ
1
0.25, 8.25 Hz, H-2 ), 4.80 (d, 1H, J = 8.0 Hz, H-1 ), 4.31 (d, 1H, J = 11.0 Hz,
ꢁ
ꢁꢁ
ꢁ
H-6 a), 4.25 (t, 1H, J = 6.75 Hz, H-5 ), 4.08 (dd, 1H, J = 12.0, 5.5 Hz, H-6 b),
ꢁ
ꢁ
4
2
(
.04 (dd, 1H, J = 6.75, 2.25 Hz, 2H, H-6 a), 4.03 (dd, 1H, J = 6.75, 2.25 Hz,
ꢁ
ꢁ
ꢁ
ꢁ
H, H-6 b), 3.91–3.84 (m, 1H, H-5 ), 3.82 (t, 1H, J = 9.25 Hz, H-4 ), 2.11–1.91
1
3
s, 21H, 7×COCH3); C NMR (DMSO-d6): δ (ppm) 178.7 (C S), 170.2–69.1
ꢁ
ꢁꢁ
ꢁꢁꢁ
ꢁꢁꢁ
(
(
7×COCH3), 141.6 (CH
N), 148.4 (C-3 ), 135.7 (C-1 ), 133.4 (C-6 ), 130.2
ꢁ
ꢁ
ꢁꢁ
ꢁꢁꢁ
ꢁꢁꢁ
ꢁꢁ
ꢁ
ꢁ
C-5 ), 124.4 (C-4 ), 121.9 (C-2 ), 99.6 (C-1 ), 81.3 (C-1 ), 76.0 (C-4 ), 73.5
ꢁ
ꢁ
ꢁꢁ
ꢁꢁ
ꢁꢁ
ꢁꢁ
(
6
8
C-5 ), 72.7 (C-3 ), 71.1 (C-2 ), 70.4 (C-3 ), 69.8 (C-5 ), 68.9 (C-2 ), 67.1 (C-4 ),
ꢁ
ꢁꢁ
+
2.4 (C-6 ), 61.0 (C-6 ), 20.7–20.3 (7×COCH3); ESI-MS: m/z: 865.06 [M+Na] ,
+
43.02 [M+H] , 823.13, 658.18, 331.21; calc. for C34H42N4O19S = 842.22 Da.
Anal. calcd. for C34H42N4O19S: C, 48.45; H, 5.02; N, 6.65; S, 3.80. Found: C,
48.49; H, 5.00; N, 6.63; S, 3.78.
2
-Nitrobenzaldehyde
ꢁ
ꢁ
ꢁ
ꢁ
N-(2,3,6,2 ,3 ,4 ,6 -hepta-O-acetyl-β-D-lactosyl)thiosemicarbazone (5d)
From 709 mg of 3 and 151 mg of 4d. Recrystallized from 96% ethanol to
yield white crystals of 5d. Yield: 99 mg, 83% (reaction time: 5 min, procedure

N-(Per-O-acetylated β-D-Lactosyl)Thiosemicarbazones
5
31
◦
25
A); 758 mg, 90% (reaction time: 3 min, procedure B); mp 170–172 C; [α] +87.2
D
–
1
(
c 1.0, CHCl3); FTIR (KBr) ν / cm : 3473, 3295, 1751, 1615, 1373, 1230, 1048;
1
H NMR (DMSO-d6): δ (ppm) 12.18 (s, 1H, NH-2), 8.77 (d, 1H, J = 9.0 Hz,
ꢁ
ꢁꢁ
NH-4), 8.53 (s, 1H, CH N), 8.06 (d, 1H, J = 8.5 Hz, H-3 ), 7.68 (t, 1H, J =
ꢁ
ꢁꢁ
ꢁꢁꢁ
ꢁꢁꢁ
7
5
3
.5 Hz, H-4 ), 7.79 (t, 1H, J = 7.5 Hz, H-5 ), 8.38 (d, 1H, J = 7.5 Hz, H-6 ),
ꢁ
ꢁ
.86 (t, 1H, J = 9.25 Hz, H-1 ), 5.30 (t, 1H, J = 9.25 Hz, H-3 ), 5.24 (d, 1H, J =
ꢁ
ꢁ
ꢁ
.5 Hz, H-4 ), 5.20 (t, 1H, J = 9.5 Hz, H-2 ), 5.15 (dd, 1H, J = 10.25, 3.75 Hz,
ꢁ
ꢁ
ꢁꢁ
ꢁꢁ
H-3 ), 4.87 (dd, 1H, J = 10.25, 8.25 Hz, H-2 ), 4.80 (d, 1H, J = 8.0 Hz, H-1 ),
ꢁ
ꢁꢁ
4
=
.30 (d, 1H, J = 12.0 Hz, H-6 a), 4.25 (t, 1H, J = 6.5 Hz, H-5 ), 4.07 (dd, 1H, J
ꢁ
ꢁꢁ
ꢁꢁ
12.25, 5.75 Hz, H-6 b), 4.05–4.02 (m, 2H, H-6 a & H-6 b), 3.92–3.89 (m, 1H,
ꢁ
ꢁ
13
H-5 ), 3.80 (t, 1H, J = 9.5 Hz, H-4 ), 2.11–1.90 (s, 21H, 7×COCH3); C NMR
ꢁ
ꢁꢁ
(
(
3
(
(
DMSO-d6): δ (ppm) 178.8 (C S), 170.2–69.0 (7×COCH3), 148.4 (C-2 ), 138.9
ꢁ
ꢁꢁ
ꢁꢁꢁ
ꢁ
ꢁꢁꢁ
ꢁꢁꢁ
CH
N), 133.4 (C-5 ), 130.8 (C-4 ), 128.3 (C-6 ), 128.1 (C-1 ), 124.6 (C-
ꢁ
ꢁꢁ
ꢁꢁ
ꢁ
ꢁ
ꢁ
ꢁ
), 99.6 (C-1 ), 81.3 (C-1 ), 76.0 (C-4 ), 73.4 (C-5 ), 72.7 (C-3 ), 71.1 (C-2 ), 70.4
ꢁ
ꢁ
ꢁꢁ
ꢁꢁ
ꢁꢁ
ꢁ
ꢁꢁ
C-3 ), 69.7 (C-5 ), 68.9 (C-2 ), 67.1 (C-4 ), 62.3 (C-6 ), 61.0 (C-6 ), 20.6–20.2
+
+
7×COCH3); ESI-MS: m/z: 865.19 [M+Na] , 843.28 [M+H] , 783.37, 658.41,
3
4
31.69; calc. for C34H42N4O19S = 842.22 Da. Anal. calcd. for C34H42N4O19S: C,
8.45; H, 5.02; N, 6.65; S, 3.80. Found: C, 47.49; H, 5.05; N, 6.68; S, 3.83.
4
-Fluorobenzaldehyde
ꢁ
ꢁ
ꢁ
ꢁ
N-(2,3,6,2 ,3 ,4 ,6 -hepta-O-acetyl-β-D-lactosyl)thiosemicarbazone (5e)
From 709 mg of 3 and 124 mg of 4e. Recrystallized from 96% ethanol to
yield white crystals of 5e. Yield: 70 mg, 70% (reaction time: 5 min, procedure
◦
25
A); 733 mg, 91% (reaction time: 2 min, procedure B); mp 159–160 C; [α] +95.4
D
–
1
(
c 1.0, CHCl3); FTIR (KBr) ν / cm : 3563, 3320, 1752, 1601, 1376, 1238, 1048;
1
H NMR (DMSO-d6): δ (ppm) 11.90 (s, 1H, NH-2), 8.66 (d, 1H, J = 9.0 Hz, NH-
ꢁ
ꢁꢁ
ꢁꢁꢁ
4
8
1
), 8.09 (s, 1H, CH
N), 7.88 (d, 2H, J = 8.5 Hz, JHF = 5.5, H-2 & H-6 ),
ꢁ
ꢁꢁ
ꢁꢁꢁ
ꢁ
.10 (d, 2H, J = 8.5 Hz, H-3 & H-5 ), 5.85 (t, 1H, J = 9.25 Hz, H-1 ), 5.30 (d,
ꢁ
ꢁꢁ
H, J = 9.25 Hz, H-3 ), 5.24 (d, 1H, J = 3.5 Hz, H-4 ), 5.19 (d, 1H, J = 9.25 Hz,
ꢁ
ꢁꢁ
H-2 ), 5.17 (dd, 1H, J = 9.75, 3.75 Hz, H-3 ), 4.88 (dd, 1H, J = 10.5, 3.25 Hz,
ꢁ
ꢁ
ꢁꢁ
ꢁ
H-2 ), 4.80 (d, 1H, J = 8.0 Hz, H-1 ), 4.31 (d, 1H, J = 11.5 Hz, H-6 a), 4.25
ꢁ
ꢁ
ꢁ
(
t, 1H, J = 6.5 Hz, H-5 ), 4.07 (dd, 1H, J = 5.5, 11.0 Hz, H-6 b), 4.04–4.02 (m,
ꢁ
ꢁ
ꢁꢁ
ꢁ
ꢁ
2
2
1
H, H-6 a & H-6 b), 3.90–3.87 (m, 1H, H-5 ), 3.81 (t, 1H, J = 9.5 Hz, H-4 ),
13
.11–2.01 (s, 21H, 7×COCH3); C NMR (DMSO-d6): δ (ppm) 178.4 (C
S),
ꢁ
ꢁꢁ
70.2–169.1 (7×COCH3), 163.3 (C-4 ), 142.6 (CH N), 130.4 (JCF = 11.4 Hz,
ꢁ
ꢁꢁ
ꢁꢁꢁ
ꢁꢁꢁ
ꢁꢁꢁ
ꢁ
C-1 ), 129.7 (J = 33.5 Hz, C-2 & C-6 ), 115.7 (J = 87 Hz, C-3 & C-
CF
CF
ꢁ
ꢁꢁ
ꢁꢁ
ꢁ
ꢁ
ꢁ
ꢁ
5
(
(
), 99.6 (C-1 ), 81.3 (C-1 ), 76.0 (C-4 ), 73.4 (C-5 ), 72.7 (C-3 ), 71.1 (C-2 ), 70.4
ꢁ
ꢁ
ꢁꢁ
ꢁꢁ
ꢁꢁ
ꢁ
ꢁꢁ
C-3 ), 69.8 (C-5 ), 68.9 (C-2 ), 67.1 (C-4 ), 62.4 (C-6 ), 61.0 (C-6 ), 20.7–20.2
+
+
7×COCH3); ESI-MS: m/z: 816.07 [M+H] , 838.12 [M+Na] , 784.41, 756.21,
40.29, 598.31, 331.18; calc. for C34H42FN3O17S = 815.22 Da. Anal. calcd. for
C34H42FN3O17S: C, 50.06; H, 5.19; N, 5.15; S, 3.93. Found: C, 50.10; H, 5.16; N,
.18; S, 3.91.
6
5

32 N. Dinh Thanh et al.
-Chlorobenzaldehyde
5
4
ꢁ
ꢁ
ꢁ
ꢁ
N-(2,3,6,2 ,3 ,4 ,6 -hepta-O-acetyl-β-D-lactosyl)thiosemicarbazone (5f)
From 709 mg of 3 and 140 mg of 4f. Recrystallized from 96% ethanol to
yield white crystals of 5f. Yield: 632 mg, 76% (reaction time: 5 min, proce-
◦
25
D
dure A); 773 mg, 93% (reaction time: 2 min, procedure B); mp 189–190 C; [α]
–
1
+
100.2 (c 1.0, CHCl3); FTIR (KBr) ν / cm : 3279, 3215, 1751, 1605, 1371, 1222,
1
1
046; H NMR (DMSO-d6): δ (ppm) 11.95 (s, 1H, NH-2), 8.70 (d, 1H, J = 9.0 Hz,
ꢁ
ꢁꢁ
ꢁꢁꢁ
NH-4), 8.08 (s, 1H, CH N), 7.85 (d, 2H, J = 8.5 Hz, H-2 & H-6 ), 7.49 (d,
ꢁ
ꢁꢁ
ꢁꢁꢁ
ꢁ
2
9
5
1
H, J = 8.5 Hz, H-3 & H-5 ), 5.85 (t, 1H, J = 9.0 Hz, H-1 ), 5.30 (t, 1H, J =
ꢁ
ꢁꢁ
ꢁ
.25 Hz, H-3 ), 5.24 (d, 1H, J = 3.5 Hz, H-4 ), 5.20 (t, 1H, J = 9.25 Hz, H-2 ),
ꢁ
ꢁ
ꢁꢁ
.16 (dd, 1H, J = 3.5, 10.0 Hz, H-3 ), 4.88 (t, 1H, J = 9.0 Hz, H-2 ), 4.80 (d,
ꢁ
ꢁ
ꢁ
H, J = 7.5 Hz, H-1 ), 4.30 (d, 1H, J = 11.5 Hz, H-6 a), 4.25 (t, 1H, J = 6.0 Hz,
ꢁ
ꢁ
ꢁ
ꢁꢁ
ꢁꢁ
H-5 ), 4.08–4.05 (m, 1H, H-6 b), 4.04–4.03 (m, 2H, H-6 a & H-6 b), 3.90–3.88
ꢁ
ꢁ
(
m, 1H, H-5 ), 3.80 (t, 1H, J = 9.5 Hz, H-4 ), 2.11–1.91 (s, 21H, 7×COCH3);
1
3
C NMR (DMSO-d6): δ (ppm) 178.3 (C
S), 170.5–169.4 (7×COCH3), 142.7
ꢁ
ꢁꢁ
ꢁꢁꢁ
ꢁꢁꢁ
ꢁꢁꢁ
ꢁꢁꢁ
(
CH
N), 134.9 (C-4 ), 132.5 (C-1 ), 129.8 (C-2 & C-6 ), 128.8 (C-3
&
ꢁ
ꢁꢁ
ꢁꢁ
ꢁ
ꢁ
ꢁ
ꢁ
C-5 ), 99.6 (C-1 ), 81.2 (C-1 ), 75.9 (C-4 ), 73.5 (C-5 ), 72.6 (C-3 ), 70.9 (C-
ꢁ
ꢁꢁ
ꢁꢁ
ꢁꢁ
ꢁꢁ
ꢁ
+
ꢁꢁ
+
2
2
8
), 70.3 (C-3 ), 69.7 (C-5 ), 67.0 (C-4 ), 68.8 (C-2 ), 62.2 (C-6 ), 60.9 (C-6 ),
0.5–20.1 (7×COCH3); ESI-MS: m/z: 854.17/856.17, [M+Na] ]/[M+2+Na] ,
+
+
37
32.13/834.09 [M+H] ]/[M+2+H] , 790.18, 656.27, 640.38, 598.39, 331.30;
3
5
calc. for C34H42 ClN3O17S/C34H42 ClN3O17S = 831.19/833.19 Da. Anal. calcd.
for C34H42ClN3O17S: C, 49.07; H, 5.09; N, 5.05; S, 3.85. Found: C, 49.11; H, 5.06;
N, 5.08; S, 3.90.
3
-Chlorobenzaldehyde
ꢁ
ꢁ
ꢁ
ꢁ
N-(2,3,6,2 ,3 ,4 ,6 -hepta-O-acetyl-β-D-lactosyl)thiosemicarbazone (5g)
From 709 mg of 3 and 140 mg of 4g. Recrystallized from 96% ethanol
to yield white crystals of 5g. Yield: 565 mg, 68% (reaction time: 5 min, pro-
◦
cedure A); 806 mg, 97% (reaction time: 3 min, procedure B); mp 152–153 C;
2
5
–1
[
1
α] +88.7 (c 1.0, CHCl3); FTIR (KBr) ν / cm : 3500, 3336, 1751, 1605, 1371,
D
1
222, 1046; H NMR (DMSO-d6): δ (ppm) 11.99 (s, 1H, NH-2), 8.75 (d, 1H,
ꢁ
ꢁꢁ
J = 9.0 Hz, NH-4), 8.08 (s, 1H, CH
N), 7.98 (s, H-2 ), 7.48–7.44 (m, 1H,
ꢁ
ꢁꢁ
ꢁꢁꢁ
ꢁꢁꢁ
H-4 ), 7.48–7.44 (m, 1H, H-5 ), 7.70 (dd, 1H, J = 7.0, 1.5 Hz, H-6 ), 5.85
ꢁ
ꢁ
(
t, 1H, J = 9.0 Hz, H-1 ), 5.30 (t, 1H, J = 9.0 Hz, H-3 ), 5.24 (d, 1H, J =
ꢁ
ꢁ
ꢁ
3
.5 Hz, H-4 ), 5.21 (t, 1H, J = 9.25 Hz, H-2 ), 5.16 (dd, 1H, J = 3.5, 10.0 Hz,
ꢁ
ꢁ
ꢁꢁ
ꢁꢁ
H-3 ), 4.88 (dd, 1H, J = 2.0, 8.0 Hz, H-2 ), 4.80 (d, 1H, J = 8.0 Hz, H-1 ),
ꢁ
ꢁꢁ
4
1
1
2
.31 (d, 1H, J = 11.0 Hz, H-6 a), 4.25 (t, 1H, J = 6.75 Hz, H-5 ), 4.07 (dd,
ꢁ
ꢁꢁ
ꢁꢁ
H, J = 5.75, 12.25 Hz, H-6 b), 4.04–4.00 (m, 2H, H-6 a & H-6 b), 3.89 (ddd,
ꢁ
ꢁ
H, J = 1.5, 5.5, 10.0 Hz, H-5 ), 3.81 (t, 1H, J = 9.5 Hz, H-4 ), 2.11–1.90 (s,
1
3
1H, 7×COCH3); C NMR (DMSO-d6): δ (ppm) 178.6 (C
S), 170.1–169.0
ꢁ
ꢁꢁ
ꢁꢁꢁ
ꢁꢁꢁ
(
(
7×COCH3), 142.2 (CH
N), 135.9 (C-1 ), 133.7 (C-3 ), 130.5 (C-4 ), 129.8
ꢁ
ꢁꢁ
ꢁꢁꢁ
ꢁꢁꢁ
ꢁꢁ
ꢁ
ꢁ
C-5 ), 126.7 (C-6 ), 126.3 (C-2 ), 99.6 (C-1 ), 81.3 (C-1 ), 76.0 (C-4 ), 73.4

N-(Per-O-acetylated β-D-Lactosyl)Thiosemicarbazones
5
33
ꢁ
ꢁ
ꢁ
ꢁꢁ
ꢁꢁ
ꢁꢁ
ꢁꢁ
(
6
[
C-5 ), 72.7 (C-3 ), 71.1 (C-2 ), 70.4 (C-3 ), 69.7 (C-5 ), 68.9 (C-2 ), 67.1 (C-4 ),
ꢁ
ꢁꢁ
2.3 (C-6 ), 61.0 (C-6 ), 20.8–20.2 (7×COCH3); ESI-MS: m/z: 854.22/856.12
+
+
M+Na] ]/[M+2+Na] , 794.36, 732.34, 658.33, 561.70, 473.73, 331.64; calc.
35 37
for C34H42 ClN3O17S/C34H42 ClN3O17S = 831.19/833.19 Da. Anal. calcd. for
C34H42ClN3O17S: C, 49.07; H, 5.09; N, 5.05; S, 3.85. Found: C, 49.05; H, 5.12;
N, 5.03; S, 3.83.
2
-Chlorobenzaldehyde
ꢁ
ꢁ
ꢁ
ꢁ
N-(2,3,6,2 ,3 ,4 ,6 -hepta-O-acetyl-β-D-lactosyl)thiosemicarbazone (5h)
From 709 mg of 3 and 140 mg of 4h. Recrystallized from 96% ethanol to
yield white crystals of 5h. Yield: 648 mg, 78% (reaction time: 5 min, proce-
◦
25
D
dure A); 765 mg, 92% (reaction time: 3 min, procedure B); mp 187–188 C; [α]
–
1
+
87.2 (c 1.0, CHCl3); FTIR (KBr) ν / cm : 3309, 1750, 1615, 1375, 1223, 1050;
1
H NMR (DMSO-d6): δ (ppm) 12.08 (s, 1H, NH-2), 8.75 (d, 1H, J = 9.0 Hz,
ꢁ
ꢁꢁꢁ
ꢁꢁ
NH-4), 8.52 (s, 1H, CH
N), 7.51 (dd, 1H, J = 8.0, 0.5 Hz, H-3 ), 7.44 (td,
ꢁ
ꢁꢁ
ꢁꢁ
1
1
9
2
H, J = 7.25, 2.0 Hz, H-4 ), 7.40 (t, 1H, J = 7.25 ), Hz, H-5 ), 8.27 (dd,
ꢁ
ꢁꢁ
ꢁ
H, J = 7.5, 1.5 Hz, H-6 ), 5.86 (t, 1H, J = 9.0 Hz, H-1 ), 5.30 (t, 1H, J =
ꢁ
ꢁꢁ
.25 Hz, H-3 ), 5.24 (d, 1H, J = 3.0 Hz, H-4 ), 5.23 (t, 1H, J = 9.25 Hz, H-
ꢁ
ꢁꢁ
), 5.15 (dd, 1H, J = 10.0, 3.5 Hz, H-3 ), 4.87 (dd, 1H, J = 10.0, 3.0 Hz,
ꢁ
ꢁ
ꢁꢁ
ꢁ
H-2 ), 4.80 (d, 1H, J = 8.0 Hz, H-1 ), 4.33–4.30 (m, 1H, H-6 a), 4.25 (t, 1H,
ꢁ
ꢁ
ꢁ
J = 6.75 Hz, H-5 ), 4.06 (dd, 1H, J = 5.25, 12.25 Hz, H-6 b), 4.03–4.02 (m,
ꢁ
ꢁꢁ
ꢁ
ꢁ
2
2
1
H, H-6 a & H-6 b), 3.90–3.88 (m, 1H, H-5 ), 3.80 (t, 1H, J = 9.5 Hz, H-4 ),
13
.11–1.92 (s, 21H, 7×COCH3); C NMR (DMSO-d6): δ (ppm) 179.1 (C
S),
ꢁ
ꢁꢁ
ꢁꢁꢁ
70.7–169.5 (7×COCH3), 140.2 (CH
N), 133.9 (C-1 ), 132.1 (C-2 ), 131.6
C-4 ), 130.3 (C-3 ), 127.8 (C-5 ), 128.0 (C-6 ), 100.1 (C-1 ), 81.7 (C-1 ), 76.5
C-4 ), 73.9 (C-5 ), 73.2 (C-3 ), 71.6 (C-2 ), 70.9 (C-3 ), 70.2 (C-5 ), 69.3 (C-
ꢁ
ꢁ
ꢁꢁ
ꢁꢁꢁ
ꢁꢁꢁ
ꢁꢁꢁ
ꢁꢁ
ꢁ
(
ꢁ
ꢁ
ꢁ
ꢁꢁ
ꢁꢁ
(
2
8
ꢁ
ꢁ
ꢁꢁ
ꢁ
ꢁꢁ
), 67.6 (C-4 ), 62.8 (C-6 ), 61.5 (C-6 ), 21.2–20.8 (7×COCH3); ESI-MS: m/z:
+
+
37
32.21/834.09 [M+H] ]/[M+2+H] , 790.17, 656.35, 640.38, 598.39, 331.28;
3
5
calc. for C34H42 ClN3O17S/C34H42 ClN3O17S = 831.19/833.19 Da. Anal. calcd.
for C34H42ClN3O17S: C, 49.07; H, 5.09; N, 5.05; S, 3.85. Found: C, 49.11; H, 5.05;
N, 5.03; S, 3.88.
4
-Bromobenzaldehyde
ꢁ
ꢁ
ꢁ
ꢁ
N-(2,3,6,2 ,3 ,4 ,6 -hepta-O-acetyl-β-D-lactosyl)thiosemicarbazone (5i)
From 709 mg of 3 and 185 mg of 4i. Recrystallized from 96% ethanol to
yield white crystals of 5i. Yield: 683 mg, 78% (reaction time: 5 min, proce-
◦
25
D
dure A); 850 mg, 97% (reaction time: 2 min, procedure B); mp 155–156 C; [α]
–
1
+
78.9 (c 1.0, CHCl3); FTIR (KBr) ν / cm : 3334, 1749, 1605, 1367, 1231, 1064;
1
H NMR (DMSO-d6): δ (ppm) 12.00 (s, 1H, NH-2), 8.70 (d, 1H, J = 9.0 Hz,
ꢁ
ꢁꢁ
ꢁꢁꢁ
NH-4), 8.08 (s, 1H, CH N), 7.82 (d, 2H, J = 8.5 Hz, H-2 & H-6 ), 7.66 (d,
ꢁ
ꢁꢁ
ꢁꢁꢁ
ꢁ
2
9
5
H, J = 8.5 Hz, H-3 & H-5 ), 5.90 (t, 1H, J = 9.0 Hz, H-1 ), 5.24 (t, 1H, J =
ꢁ
ꢁ
ꢁꢁ
ꢁꢁ
.25 Hz, H-2 ), 5.33 (t, 1H, J = 9.25 Hz, H-3 ), 5.28 (d, 1H, J = 3.5 Hz, H-4 ),
ꢁ
ꢁ
.16 (dd, 1H, J = 10.25, 3.75 Hz, H-3 ), 4.91 (dd, 1H, J = 11.5, 3.0 Hz, H-2 ),

34 N. Dinh Thanh et al.
5
ꢁ
ꢁ
ꢁ
4
6
3
1
.83 (d, 1H, J = 8.0 Hz, H-1 ), 4.33 (d, 1H, J = 11.0 Hz, H-6 a), 4.29 (t, 1H, J =
ꢁ
ꢁ
ꢁ
ꢁꢁ
.5 Hz, H-5 ), 4.09 (dd, 1H, J = 12.0, 5.5 Hz, H-6 b), 4.07–4.05 (m, 1H, H-6 a),
ꢁ
ꢁ
ꢁ
.94–3.91 (m, 1H, H-6 b), 3.93 (ddd, 1H, J = 9.75, 5.75, 1.75 Hz, H-5 ), 3.84 (t,
ꢁ
13
H, J = 9.75 Hz, H-4 ), 2.15–1.94 (s, 21H, 7×COCH3); C NMR (DMSO-d6):
δ (ppm) 178.4 (C
S), 170.2–169.0 (7×COCH3), 142.5 (CH
N), 133.0 (C-
ꢁ
ꢁꢁ
ꢁꢁꢁ
ꢁꢁꢁ
ꢁꢁꢁ
ꢁꢁꢁ
ꢁꢁꢁ
ꢁꢁ
1
), 131.7 (C-3 & C-5 ), 129.4 (C-2 & C-6 ), 123.5 (C-4 ), 99.6 (C-1 ), 81.2
ꢁ
ꢁ
ꢁ
ꢁ
ꢁ
ꢁꢁ
ꢁꢁ
(
6
C-1 ), 76.0 (C-4 ), 73.4 (C-5 ), 72.7 (C-3 ), 71.1 (C-2 ), 70.4 (C-3 ), 69.7 (C-5 ),
ꢁ
ꢁ
ꢁꢁ
ꢁ
ꢁꢁ
8.8 (C-2 ), 67.1 (C-4 ), 62.4 (C-6 ), 60.9 (C-6 ), 20.7–20.4 (7×COCH3); ESI-
+
+
MS: m/z: 876.41/877.98 [M+H] ]/[M+2+H] , 850.47, 828.21, 640.43, 598.38,
31.59; calc. for C34H4279BrN3O17S/C34H42 BrN3O17S = 875.14/877.14 Da.
81
3
Anal. calcd. for C34H42BrN3O17S: C, 46.58; H, 4.83; N, 4.79; S, 3.66. Found:
C, 46.55; H, 4.87; N, 4.82; S, 3.70.
4
-Methylbenzaldehyde
ꢁ
ꢁ
ꢁ
ꢁ
N-(2,3,6,2 ,3 ,4 ,6 -hepta-O-acetyl-β-D-lactosyl)thiosemicarbazone (5j)
From 709 mg of 3 and 120 mg of 4j. Recrystallized from 96% ethanol to
yield white crystals of 5j. Yield: 632 mg, 78% (reaction time: 5 min, procedure
◦
25
A); 786 mg, 97% (reaction time: 2 min, procedure B); mp 177–178 C; [α] +94.5
D
–
1
1
(
c 1.0, CHCl3); FTIR (KBr) ν / cm : 3212, 1748, 1615, 1373, 1242, 1046; H
NMR (DMSO-d6): δ (ppm) 11.86 (s, 1H, NH-2), 8.59 (d, 1H, J = 9.5 Hz, NH-4),
ꢁ
ꢁꢁ
ꢁ
ꢁꢁꢁ
8
8
.06 (s, 1H, CH N), 7.69 (d, 2H, J = 8.5 Hz, H-2 & H-6 ), 7.25 (d, 2H, J =
ꢁ
ꢁꢁ
ꢁꢁꢁ
.5 Hz, H-3 & H-5 ), 5.84 (t, 1H, J = 9.0 Hz, H-1 ), 5.30 (t, 1H, J = 9.25 Hz,
ꢁ
ꢁꢁ
ꢁ
H-3 ), 5.24 (d, 1H, J = 3.5 Hz, H-4 ), 5.21 (t, 1H, J = 9.25 Hz, H-2 ), 5.15 (dd,
ꢁ
ꢁ
ꢁꢁ
1
=
H, J = 10.25, 3.75 Hz, H-3 ), 4.88 (t, 1H, J = 9.0 Hz, H-2 ), 4.80 (d, 1H, J
ꢁ
ꢁ
ꢁ
8.0 Hz, H-1 ), 4.30 (d, 1H, J = 11.0 Hz, H-6 a), 4.25 (t, 1H, J = 6.5 Hz,
ꢁ
ꢁ
ꢁ
ꢁꢁ
H-5 ), 4.07 (dd, 1H, J = 12.25, 5.75 Hz, H-6 b), 4.04–4.03 (m, 2H, H-6 a &
ꢁ
ꢁ
ꢁ
ꢁ
H-6 b), 3.90–3.87 (m, 1H, H-5 ), 3.81 (t, 1H, J = 9.5 Hz, H-4 ), 2.34 (s, 3H,
ꢁ
ꢁꢁ
13
4
-CH3), 2.11–1.90 (s, 21H, 7×COCH3); C NMR (DMSO-d6): δ (ppm) 178.2
ꢁ
ꢁꢁ
ꢁꢁꢁ
(
C
S), 170.2–169.0 (7×COCH3), 143.9 (CH N), 140.2 (C-4 ), 131.0 (C-1 ),
ꢁ
ꢁꢁ
ꢁꢁꢁ
ꢁꢁꢁ
ꢁꢁꢁ
ꢁꢁ
ꢁ
ꢁ
1
7
(
29.3 (C-3 & C-5 ), 127.5 (C-2 & C-6 ), 99.6 (C-1 ), 81.2 (C-1 ), 76.0 (C-4 ),
ꢁ
ꢁ
ꢁ
ꢁꢁ
ꢁꢁ
ꢁꢁ
3.4 (C-5 ), 72.7 (C-3 ), 71.1 (C-2 ), 70.4 (C-3 ), 69.7 (C-5 ), 68.9 (C-2 ), 67.1
ꢁ
ꢁ
ꢁ
ꢁꢁ
ꢁꢁꢁ
C-4 ), 62.3 (C-6 ), 60.9 (C-6 ), 20.6–20.2 (7×COCH3), 21.0 (4 -CH3); ESI-MS:
+
+
m/z: 812.25 [M+H] , 834.31 [M+Na] , 770.31, 640.45, 598.44, 331.34; calc. for
C35H45N3O17S = 811.25 Da. Anal. calcd. for C35H45N3O17S: C, 51.78; H, 5.59;
N, 5.18; S, 3.95. Found: C, 51.82; H, 5.56; N, 5.16; S, 3.99.
4
-Isopropylbenzaldehyde
ꢁ
ꢁ
ꢁ
ꢁ
N-(2,3,6,2 ,3 ,4 ,6 -hepta-O-acetyl-β-D-lactosyl)thiosemicarbazone (5k)
From 709 mg of 3 and 148 mg of 4k. Recrystallized from 96% ethanol to
yield white crystals of 5k. Yield: 604 mg, 72% (reaction time: 5 min, procedure
◦
25
A); 814 mg, 97% (reaction time: 2 min, procedure B); mp 148–149 C; [α] +95.4
D
–
1
(
c 1.0, CHCl3); FTIR (KBr) ν / cm : 3501, 3337, 1749, 1610, 1375, 1230, 1049;
H NMR (DMSO-d6): δ (ppm) 11.87 (s, 1H, NH-2), 8.57 (d, 1H, J = 9.0 Hz,
1

N-(Per-O-acetylated β-D-Lactosyl)Thiosemicarbazones
5
35
ꢁ
ꢁꢁ
ꢁꢁꢁ
NH-4), 8.09 (s, 1H, CH N), 7.72 (d, 2H, J = 8.0 Hz, H-2 & H-6 ), 7.30 (d,
ꢁ
ꢁꢁ
ꢁꢁꢁ
ꢁ
2
H, J = 8.0 Hz, H-3 & H-5 ), 5.83 (t, 1H, J = 9.25 Hz, H-1 ), 5.30 (t, 1H,
ꢁ
ꢁꢁ
J = 9.25 Hz, H-3 ), 5.24 (d, 1H, J = 3.5 Hz, H-4 ), 5.18 (t, 1H, J = 9.5 Hz,
ꢁ
ꢁꢁ
ꢁꢁ
H-2 ), 5.15 (dd, 1H, J = 10.5, 3.75 Hz, H-3 ), 4.88 (t, 1H, J = 9.75 Hz, H-2 ),
ꢁ
ꢁ
ꢁ
4
=
.80 (d, 1H, J = 8.0 Hz, H-1 ), 4.30 (d, 1H, J = 11.5 Hz, H-6 a), 4.25 (t, 1H, J
ꢁ
ꢁ
ꢁ
ꢁꢁ
ꢁꢁ
5.75 Hz, H-5 ), 4.09–4.03 (m, 3H, H-6 b, H-6 a & H-6 b), 3.88–3.87 (m, 1H,
ꢁ
ꢁ
ꢁꢁꢁ
H-5 ), 3.81 (t, 1H, J = 9.25 Hz, H-4 ), 2.92 [q, 1H, J = 7.0 Hz, 4 -CH(CH3)2],
ꢁ
ꢁꢁ
13
2
.11–1.90 (s, 21H, 7×COCH3), 1.21 [d, 6H, J = 7.0 Hz, 4 -CH(CH3)2];
C
NMR (DMSO-d6): δ (ppm) 178.2 (C
S), 170.2–169.0 (7×COCH3), 150.9 (C-
ꢁ
ꢁ
ꢁꢁ
ꢁꢁ
ꢁꢁꢁ
ꢁꢁꢁ
ꢁ
ꢁꢁꢁ
ꢁꢁꢁ
ꢁ
4
5
(
), 143.8 (CH
N), 131.4 (C-1 ), 127.6 (C-2 & C-6 ), 126.6 (C-3 & C-
ꢁꢁ
ꢁ
ꢁ
ꢁ
), 99.6 (C-1 ), 81.1 (C-1 ), 76.0 (C-4 ), 73.4 (C-5 ), 72.7 (C-3 ), 71.0 (C-2 ), 70.3
C-3 ), 69.7 (C-5 ), 68.8 (C-2 ), 67.1 (C-4 ), 62.3 (C-6 ), 60.9 (C-6 ), 33.4 [4 -
ꢁ
ꢁ
ꢁꢁ
ꢁꢁ
ꢁꢁ
ꢁ
ꢁꢁ
ꢁꢁ
ꢁ
ꢁꢁ
CH(CH3)2], 23.5 [4 -CH(CH3)2], 20.6–20.2 (7×COCH3); ESI-MS: m/z: 862.41
+
[
M+Na] , 848.66, 820.68, 732.72, 596.04, 351.28; calc. for C37H49N3O17S =
8
39.28 Da. Anal. calcd. for C37H49N3O17S: C, 52.91; H, 5.88; N, 5.00; S, 3.82.
Found: C, 52.95; H, 5.85; N, 5.03; S, 3.86.
4
-Methoxybenzaldehyde
ꢁ
ꢁ
ꢁ
ꢁ
N-(2,3,6,2 ,3 ,4 ,6 -hepta-O-acetyl-β-D-lactosyl)thiosemicarbazone (5l)
From 709 mg of 3 and 136 mg of 4l. Recrystallized from 96% ethanol to
yield white crystals of 5l. Yield: 645 mg, 78% (reaction time: 5 min, procedure
◦
25
A); 802 mg, 97% (reaction time: 2 min, procedure B); mp 198–199 C; [α] +97.3
D
–
1
(
c 1.0, CHCl3); FTIR (KBr) ν / cm : 3348, 3284, 1737, 1604, 1370, 1228, 1044;
1
H NMR (DMSO-d6): δ (ppm) 11.81 (s, 1H, NH-2), 8.55 (d, 1H, J = 9.0 Hz,
ꢁ
ꢁꢁ
ꢁꢁꢁ
NH-4), 8.05 (s, 1H, CH N), 7.75 (d, 2H, J = 8.5 Hz, H-2 & H-6 ), 6.99 (d,
ꢁ
ꢁꢁ
ꢁꢁꢁ
ꢁ
2
H, J = 8.5 Hz, H-3 & H-5 ), 5.83 (t, 1H, J = 9.25 Hz, H-1 ), 5.30 (t, 1H,
ꢁ
ꢁꢁ
J = 9.0 Hz, H-3 ), 5.24 (d, 1H, J = 3.0 Hz, H-4 ), 5.18 (t, 1H, J = 9.25 Hz,
ꢁ
ꢁꢁ
ꢁꢁ
H-2 ), 5.15 (dd, 1H, J = 10.0, 3.0 Hz, H-3 ), 4.88 (t, 1H, J = 9.25 Hz, H-2 ),
ꢁ
ꢁ
ꢁ
4
.80 (d, 1H, J = 7.5 Hz, H-1 ), 4.30 (d, 1H, J = 11.5 Hz, H-6 a), 4.24 (t, 1H,
ꢁ
ꢁ
ꢁ
ꢁꢁ
ꢁꢁꢁ
J = 6.75 Hz, H-5 ), 4.09–4.05 (m, 1H, H-6 b), 4.04–4.03 (m, 2H, H-6 a & H-
ꢁ
ꢁ
ꢁ
ꢁ
6
2
1
1
b), 3.89–3.87 (m, 1H, H-4 ), 3.82–3.79 (m, 1H, H-5 ), 3.81 (s, 3H, 4 -OMe),
1
3
.11–1.91 (s, 21H, 7×COCH3); C NMR (DMSO-d6): δ (ppm) 177.8 (C
S),
ꢁ
ꢁꢁ
ꢁꢁꢁ
ꢁꢁꢁ
70.5–169.4 (7×COCH3), 161.1 (C-4 ), 144.1 (CH N), 129.2 (C-2 & C-6 ),
ꢁ
ꢁꢁ
ꢁꢁꢁ
ꢁꢁꢁ
ꢁꢁ
ꢁ
ꢁ
26.0 (C-1 ), 114.2 (C-3 & C-5 ), 99.6 (C-1 ), 81.1 (C-1 ), 75.9 (C-4 ), 73.5
ꢁ
ꢁ
ꢁ
ꢁꢁ
ꢁꢁ
ꢁꢁ
ꢁꢁ
(
C-5 ), 72.6 (C-3 ), 70.9 (C-2 ), 70.3 (C-3 ), 69.7 (C-5 ), 68.8 (C-2 ), 67.1 (C-4 ),
ꢁ
ꢁꢁ
ꢁꢁꢁ
6
8
2.2 (C-6 ), 60.9 (C-6 ), 20.5–20.1 (7×COCH3), 55.2 (4 -OCH3); ESI-MS: m/z:
+
+
50.28 [M+Na] , 828.23 [M+H] , 786.26, 768.35, 640.38, 540.36, 331.27; calc.
for C35H45N3O18S = 827.24 Da. Anal. calcd. for C35H45N3O18S: C, 50.78; H,
5.48; N, 5.08; S, 3.87. Found: C, 50.82; H, 5.45; N, 5.12; S, 3.85.
3
-Methoxybenzaldehyde
ꢁ
ꢁ
ꢁ
ꢁ
N-(2,3,6,2 ,3 ,4 ,6 -hepta-O-acetyl-β-D-lactosyl)thiosemicarbazone (5m)
From 709 mg of 3 and 136 mg of 4m. Recrystallized from 96% ethanol to
yield white crystals of 5m. Yield: 604 mg, 73% (reaction time: 5 min, procedure

36 N. Dinh Thanh et al.
5
◦
25
A); 761 mg, 92% (reaction time: 2 min, procedure B); mp 196–197 C; [α] +95.8
D
–
1
(
c 1.0, CHCl3); FTIR (KBr) ν / cm : 3345, 3160, 1747, 1605, 1376, 1241, 1042;
1
H NMR (DMSO-d6): δ (ppm) 11.95 (s, 1H, NH-2), 8.62 (d, 1H, J = 9.0 Hz, NH-
ꢁ
ꢁꢁ
ꢁꢁꢁ
4
(
), 8.07 (s, 1H, CH N), 7.44 (s, H-2 ), 7.34 (d, 1H, J = 8.0 Hz, H-5 ), 7.31
ꢁ
ꢁꢁ
ꢁꢁꢁ
d, 1H, J = 8.0 Hz, H-6 ), 7.00 (dd, 1H, J = 8.0, 1.5 Hz, H-4 ), 5.79 (t, 1H, J
ꢁ
ꢁ
ꢁ
=
9.0 Hz, H-1 ), 5.31 (t, 1H, J = 9.25 Hz, H-3 ), 5.18 (t, 1H, J = 9.25 Hz, H-2 ),
ꢁ
ꢁ
ꢁꢁ
5
1
1
.24 (d, 1H, J = 3.5 Hz, H-4 ), 5.15 (dd, 1H, J = 10.5, 3.5 Hz, H-3 ), 4.88 (dd,
ꢁ
ꢁ
ꢁꢁ
H, J = 10.0, 3.0 Hz, H-2 ), 4.79 (d, 1H, J = 7.5 Hz, H-1 ), 4.30 (d, 1H, J =
ꢁ
ꢁꢁ
1.0 Hz, H-6 a), 4.24 (t, 1H, J = 6.75 Hz, H-5 ), 4.07 (dd, 1H, J = 12.5, 6.75 Hz,
ꢁ
ꢁꢁ
ꢁꢁ
H-6 b), 4.04 (d, 1H, J = 6.5 Hz, H-6 a), 4.03 (d, 1H, J = 6.5 Hz, H-6 b), 3.88
ꢁ
ꢁꢁꢁ
(
ddd, 1H, J = 10.0, 5.5, 3.5 Hz, H-5 ), 3.83 (s, 3H, 3 -OCH3), 3.81 (t, 1H, J
ꢁ
13
=
9.0 Hz, H-4 ), 2.11–2.01 (s, 21H, 7×COCH3); C NMR (DMSO-d6): δ (ppm)
ꢁ
ꢁꢁ
1
78.4 (C
S), 170.2–169.0 (7×COCH3), 159.6 (C-3 ), 143.5 (CH
N), 135.1
ꢁ
ꢁꢁ
ꢁꢁꢁ
ꢁꢁꢁ
ꢁꢁꢁ
ꢁꢁꢁ
ꢁꢁ
(
(
6
(
C-1 ), 129.6 (C-5 ), 120.7 (C-6 ), 116.5 (C-4 ), 111.4 (C-2 ), 99.6 (C-1 ), 81.2
ꢁ
ꢁ
ꢁ
ꢁ
ꢁ
ꢁꢁ
ꢁꢁ
C-1 ), 76.1 (C-4 ), 73.4 (C-5 ), 72.5 (C-3 ), 70.9 (C-2 ), 70.4 (C-3 ), 69.7 (C-5 ),
ꢁ
ꢁ
ꢁꢁ
ꢁ
ꢁꢁ
8.8 (C-2 ), 67.1 (C-4 ), 62.3 (C-6 ), 60.9 (C-6 ), 20.6–20.2 (7×COCH3), 55.2
ꢁ
ꢁ
+
+
3 -OCH3); ESI-MS: m/z: 850.28 [M+Na] , 828.27 [M+H] , 786.33, 744.50,
56.21, 445.47, 331.37; calc. for C35H45N3O18S = 827.24 Da. Anal. calcd. for
C35H45N3O18S: C, 50.78; H, 5.48; N, 5.08; S, 3.87. Found: C, 50.76; H, 5.50; N,
6
5.12; S, 3.91.
2
-Methoxybenzaldehyde
ꢁ
ꢁ
ꢁ
ꢁ
N-(2,3,6,2 ,3 ,4 ,6 -hepta-O-acetyl-β-D-lactosyl)thiosemicarbazone (5n)
From 709 mg of 3 and 136 mg of 4n. Recrystallized from 96% ethanol to
yield white crystals of 5n. Yield: 604 mg, 73% (reaction time: 5 min, procedure
◦
25
A); 802 mg, 97% (reaction time: 2 min, procedure B); mp 153–155 C; [α] +94.8
D
–
1
(
c 1.0, CHCl3); FTIR (KBr) ν / cm : 3548, 3282, 1750, 1607, 1372, 1241, 1054;
1
H NMR (DMSO-d6): δ (ppm) 11.89 (s, 1H, NH-2), 8.58 (d, 1H, J = 9.0 Hz, NH-
ꢁ
ꢁꢁ
4
), 8.45 (s, 1H, CH N), 8.06 (d, 1H, J = 8.0 Hz, H-6 ), 7.08 (d, 1H, J = 8.5 Hz,
ꢁ
ꢁꢁ
ꢁꢁꢁ
ꢁꢁꢁ
H-3 ), 7.42 (t, 1H, J = 7.25 Hz, H-4 ), 6.99 (t, 1H, J = 7.5 Hz, H-5 ), 5.84 (t,
ꢁ
ꢁ
1
4
1
H, J = 9.0 Hz, H-1 ), 5.30 (t, 1H, J = 9.25 Hz, H-3 ), 5.24 (d, 1H, J = 3.0 Hz, H-
ꢁ
ꢁ
ꢁ
ꢁꢁ
), 5.18 (t, 1H, J = 9.0 Hz, H-2 ), 5.16 (dd, 1H, J = 10.0, 3.5 Hz, H-3 ), 4.88 (t,
ꢁ
ꢁ
ꢁꢁ
H, J = 9.0 Hz, H-2 ), 4.79 (d, 1H, J = 8.0 Hz, H-1 ), 4.31 (d, 1H, J = 11.5 Hz,
ꢁ
ꢁꢁ
ꢁ
H-6 a), 4.25 (t, 1H, J = 6.25 Hz, H-5 ), 4.07 (dd, 1H, J = 12.5, 5.5 Hz, H-6 b),
ꢁ
ꢁꢁ
ꢁꢁ
3
1
.89–3.78 (m, 1H, H-5 ), 4.04–4.03 (m, 2H, H-6 a & H-6 b), 3.88 (ddd, 1H, J =
ꢁ
ꢁꢁꢁ
0.0, 5.5, 3.5 Hz, H-4 ), 3.83 (s, 3H, 2 -OCH3), 2.11–1.90 (s, 21H, 7×COCH3);
1
3
C NMR (DMSO-d6): δ (ppm) 178.2 (C
S), 170.2–169.1 (7×COCH3), 158.1
ꢁ
ꢁꢁ
ꢁꢁꢁ
ꢁꢁꢁ
ꢁꢁꢁ
(
C-3 ), 139.5 (CH
N), 131.8 (C-1 ), 126.2 (C-5 ), 121.8 (C-6 ), 120.6 (C-
ꢁ
ꢁꢁ
ꢁꢁꢁ
ꢁꢁ
ꢁ
ꢁ
ꢁ
ꢁ
2
7
6
8
), 111.8 (C-4 ), 99.6 (C-1 ), 81.2 (C-1 ), 76.1 (C-4 ), 73.4 (C-5 ), 72.7 (C-3 ),
1.1 (C-2 ), 70.4 (C-3 ), 69.8 (C-5 ), 68.9 (C-2 ), 67.1 (C-4 ), 62.4 (C-6 ), 61.0 (C-
ꢁ
ꢁꢁ
ꢁꢁ
ꢁꢁ
ꢁꢁ
ꢁ
ꢁ
ꢁ
ꢁꢁꢁ
+
), 55.7 (2 -OCH3), 20.6–20.2 (7×COCH3); ESI-MS: m/z: 850.25 [M+Na] ,
+
28.36 [M+H] ,763.47, 689.58, 615.57, 429.29, 331.72; calc. for C35H45N3O18S

N-(Per-O-acetylated β-D-Lactosyl)Thiosemicarbazones
5
37
=
827.24 Da. Anal. calcd. for C35H45N3O18S: C, 50.78; H, 5.48; N, 5.08; S, 3.87.
Found: C, 50.81; H, 5.46; N, 5.05; S, 3.86.
4
-Hydroxybenzaldehyde
ꢁ
ꢁ
ꢁ
ꢁ
N-(2,3,6,2 ,3 ,4 ,6 -hepta-O-acetyl-β-D-lactosyl)thiosemicarbazone (5o)
From 709 mg of 3 and 122 mg of 4o. Recrystallized from 96% ethanol to
yield white crystals of 5o. Yield: 634 mg, 78% (reaction time: 5 min, procedure
◦
25
A); 788 mg, 97% (reaction time: 2 min, procedure B); mp 161–162 C; [α] +84.8
D
–
1
(
c 1.0, CHCl3); FTIR (KBr) ν / cm : 3548, 3340, 3275, 1749, 1609, 1372, 1238,
1
ꢁꢁꢁ
1
042; H NMR (DMSO-d6): δ (ppm) 11.76 (s, 1H, NH-2), 9.97 (s, 1H, 4 -OH),
.51 (d, 1H, J = 9.5 Hz, NH-4), 8.00 (s, 1H, CH N), 7.64 (d, 2H, J = 8.5 Hz,
8
ꢁ
ꢁꢁ
ꢁꢁꢁ
ꢁꢁꢁ
ꢁꢁꢁ
H-3 & H-5 ), 6.81 (d, 2H, J = 8.5 Hz, H-2 & H-6 ), 5.83 (t, 1H, J = 9.25 Hz,
ꢁ
ꢁ
ꢁꢁ
H-1 ), 5.30 (t, 1H, J = 9.25 Hz, H-3 ), 5.24 (d, 1H, J = 3.5 Hz, H-4 ), 5.18 (t, 1H,
ꢁ
ꢁꢁ
J = 9.25 Hz, H-2 ), 5.16 (dd, 1H, J = 10.0, 4.0 Hz, H-3 ), 4.87 (dd, 1H, J = 10.0,
ꢁ
ꢁ
ꢁꢁ
ꢁ
3
4
.0 Hz, H-2 ), 4.80 (d, 1H, J = 8.0 Hz, H-1 ), 4.30 (d, 1H, J = 11.0 Hz, H-6 a),
ꢁ
ꢁ
ꢁ
.25 (t, 1H, J = 6.75 Hz, H-5 ), 4.06 (dd, 1H, J = 9.5, 5.5 Hz, H-6 b), 4.03–4.02
ꢁ
ꢁ
ꢁꢁ
ꢁ
(
m, 2H, H-6 a & H-6 b), 3.89–3.86 (m, 1H, H-4 ), 3.80 (t, 1H, J = 9.75 Hz, H-
ꢁ
13
5
1
1
), 2.11–1.90 (s, 21H, 7×COCH3); C NMR (DMSO-d6): δ (ppm) 177.7 (C S),
ꢁ
ꢁꢁ
ꢁꢁꢁ
ꢁꢁꢁ
70.2–169.0 (7×COCH3), 159.6 (C-4 ), 144.1 (CH N), 129.4 (C-2 & C-6 ),
ꢁ
ꢁꢁ
ꢁꢁꢁ
ꢁꢁꢁ
ꢁꢁ
ꢁ
ꢁ
24.6 (C-1 ), 115.6 (C-3 & C-5 ), 99.6 (C-1 ), 81.1 (C-1 ), 76.1 (C-4 ), 73.3
ꢁ
ꢁ
ꢁ
ꢁꢁ
ꢁꢁ
ꢁꢁ
ꢁꢁ
+
(
6
8
C-5 ), 72.7 (C-3 ), 71.1 (C-2 ), 70.3 (C-3 ), 69.7 (C-5 ), 68.8 (C-2 ), 67.1 (C-4 ),
ꢁ
ꢁꢁ
2.3 (C-6 ), 60.9 (C-6 ), 20.7–20.3 (7×COCH3); ESI-MS: m/z: 836.31 [M+Na] ,
+
14.22 [M+H] , 772.33, 640.45, 598.45, 427.05, 331.45; calc. for C34H43N3O18S
=
813.23 Da. Anal. calcd. for C34H43N3O18S: C, 50.18; H, 5.33; N, 5.16; S, 3.94.
Found: C, 50.22; H, 5.31; N, 5.19; S, 3.91.
2
-Hydroxybenzaldehyde
ꢁ
ꢁ
ꢁ
ꢁ
N-(2,3,6,2 ,3 ,4 ,6 -hepta-O-acetyl-β-D-lactosyl)thiosemicarbazone (5p)
From 709 mg of 3 and 122 mg of 4p. Recrystallized from 96% ethanol to
yield white crystals of 5p. Yield: 504 mg, 62% (reaction time: 5 min, procedure
◦
25
A); 788 mg, 97% (reaction time: 2 min, procedure B); mp 150–151 C; [α] +85.9
D
–
1
(
c 1.0, CHCl3); FTIR (KBr) ν / cm : 3602, 3446, 3282, 1741, 1609, 1372, 1228,
1
ꢁꢁꢁ
1
8
4
1
040; H NMR (DMSO-d6): δ (ppm) 11.74 (s, 1H, NH-2), 9.96 (s, 1H, 2 -OH),
.49 (d, 1H, J = 9.5 Hz, NH-4), 8.00 (s, 1H, CH N), 7.63 (d, 2H, J = 8.5 Hz, H-
ꢁ
ꢁ
ꢁꢁ
ꢁꢁꢁ
ꢁꢁꢁ
ꢁꢁꢁ
& H-6 ), 6.81 (d, 2H, J = 8.5 Hz, H-3 & H-5 ), 5.83 (t, 1H, J = 9.25 Hz, H-
ꢁ
ꢁꢁ
), 5.29 (t, 1H, J = 9.25 Hz, H-3 ), 5.24 (d, 1H, J = 3.5 Hz, H-4 ), 5.15 (dd, 1H,
ꢁ
ꢁꢁ
J = 3.75, 9.75 Hz, H-2 ), 5.11 (t, 1H, J = 9.0 Hz, H-3 ), 4.87 (dd, 1H, J = 10.0,
ꢁ
ꢁ
ꢁꢁ
ꢁ
3
4
.0 Hz, H-2 ), 4.79 (d, 1H, J = 8.0 Hz, H-1 ), 4.30 (d, 1H, J = 11.5 Hz, H-6 a),
ꢁ
ꢁ
ꢁ
.24 (t, 1H, J = 6.75 Hz, H-5 ), 4.06 (dd, 1H, J = 12.0, 5.5 Hz, H-6 b), 4.04–4.02
ꢁ
ꢁ
ꢁꢁ
ꢁ
(
4
m, 2H, H-6 a & H-6 b), 3.89–3.86 (m, 1H, H-5 ), 3.80 (t, 1H, J = 9.5 Hz, H-
ꢁ
13
), 2.11–1.90 (s, 21H, 7×COCH3); C NMR (DMSO-d6): δ (ppm) 177.8 (C
ꢁ
ꢁꢁ
ꢁꢁꢁ
S), 170.2–169.1 (7×COCH3), 159.7 (C-2 ), 144.2 (CH N), 129.4 (C-4 & C-
ꢁ
ꢁꢁ
ꢁꢁꢁ
ꢁꢁꢁ
ꢁꢁꢁ
ꢁꢁ
ꢁ
ꢁ
6
), 124.7 (C-5 ), 115.6 (C-1 & C-3 ), 99.6 (C-1 ), 81.1 (C-1 ), 76.1 (C-4 ),

38 N. Dinh Thanh et al.
5
ꢁ
ꢁ
ꢁ
ꢁꢁ
ꢁꢁ
ꢁꢁ
7
(
[
3.4 (C-5 ), 72.7 (C-3 ), 71.1 (C-2 ), 70.4 (C-3 ), 69.8 (C-5 ), 68.9 (C-2 ), 67.2
ꢁ
ꢁ
ꢁ
ꢁꢁ
C-4 ), 62.4 (C-6 ), 61.0 (C-6 ), 20.7–20.3 (7×COCH3); ESI-MS: m/z: 836.39
+
+
M+Na] , 814.50 [M+H] , 794.62, 659.65, 606.01, 561.97, 430.45, 342.45; calc.
for C34H43N3O18S = 813.23 Da. Anal. calcd. for C34H43N3O18S: C, 50.18; H,
5
.33; N, 5.16; S, 3.94. Found: C, 50.15; H, 5.31; N, 5.19; S, 3.98.
3
-Methoxy-4-hydroxybenzaldehyde
ꢁ
ꢁ
ꢁ
ꢁ
N-(2,3,6,2 ,3 ,4 ,6 -hepta-O-acetyl-β-D-lactosyl)thiosemicarbazone (5q)
From 709 mg of 3 and 152 mg of 4q. Recrystallized from 96% ethanol to
yield white crystals of 5q. Yield: 649 mg, 77% (reaction time: 5 min, proce-
◦
25
D
dure A); 817 mg, 97% (reaction time: 2 min, procedure B); mp 187–188 C; [α]
–
1
+
101.4 (c 1.0, CHCl3); FTIR (KBr) ν / cm : 3480, 3337, 1749, 1603, 1377, 1224,
1
ꢁꢁꢁ
1
8
7
1
051; H NMR (DMSO-d6): δ (ppm) 11.84 (s, 1H, NH-2), 9.57 (s, 1H, 4 -OH),
ꢁ
ꢁꢁ
.50 (d, 1H, J = 9.0 Hz, NH-4), 7.98 (s, 1H, CH N), 7.46 (d, J = 1.5 Hz, H-2 ),
ꢁ
ꢁꢁ
ꢁꢁꢁ
.10 (dd, 1H, J = 8.25, 1.5 Hz, H-6 ), 6.81 (d, 1H, J = 8.0 Hz, H-5 ), 5.74 (t,
ꢁ
ꢁ
H, J = 9.25 Hz, H-1 ), 5.33 (t, 1H, J = 9.25 Hz, H-3 ), 5.24 (d, 1H, J = 3.5 Hz,
ꢁ
ꢁ
ꢁ
ꢁꢁ
ꢁꢁ
H-4 ), 5.17–5.13 (m, 2H, H-2 & H-3 ), 4.87 (dd, 1H, J = 10.0, 3.0 Hz, H-2 ),
ꢁ
ꢁ
ꢁꢁꢁ
4
.78 (d, 1H, J = 8.0 Hz, H-1 ), 4.29 (d, 1H, J = 11.0 Hz, H-6 ), 4.25 (t, 1H, J
ꢁ
ꢁ
ꢁꢁꢁ
=
6.75 Hz, H-5 ), 4.07 (dd, 1H, J = 12.0, 5.5 Hz, H-6 ), 4.04–4.02 (m, 2H, H-
ꢁ
ꢁ
ꢁꢁ
ꢁꢁꢁ
ꢁ
ꢁ
6
a & H-6 b), 3.86 (s, 3H, 3 -OMe), 3.85–3.81 (m, 2H, H-4 & H-5 ), 2.11–1.91
1
3
(
(
(
s, 21H, 7×COCH3); C NMR (DMSO-d6): δ (ppm) 177.8 (C S), 170.2–169.0
ꢁ
ꢁꢁ
ꢁꢁꢁ
ꢁꢁꢁ
7×COCH3), 149.5 (C-4 ), 147.1 (C-3 ), 144.1 (CH N), 125.0 (C-1 ), 122.4
ꢁ
ꢁ
ꢁꢁ
ꢁꢁꢁ
ꢁꢁꢁ
ꢁꢁ
ꢁ
ꢁ
C-6 ), 115.4 (C-5 ), 111.1 (C-2 ), 99.6 (C-1 ), 81.0 (C-1 ), 76.2 (C-4 ), 73.3
ꢁ
ꢁ
ꢁꢁ
ꢁꢁ
ꢁꢁ
(
4
C-5 ), 72.4 (C-3 ), 70.8 (C-2 ), 70.4 (C-3 ), 69.7 (C-5 ), 68.8 (C-2 ), 67.1 (C-
ꢁ
ꢁ
ꢁ
ꢁꢁ
ꢁꢁꢁ
), 62.3 (C-6 ), 60.9 (C-6 ), 55.7 (3 -OCH3), 20.7–20.3 (7×COCH3); ESI-MS:
+
+
m/z: 865.88 [M+Na] , 843.79 [M+H] , 824.00, 806.09, 701.09, 659.10, 424.06,
3
4
31.02; calc. for C35H45N3O19S = 843.24 Da. Anal. calcd. for C35H45N3O19S: C,
9.82; H, 5.38; N, 4.98; S, 3.80. Found: C, 49.86; H, 5.35; N, 4.96; S, 3.83.
3
-Ethoxy-4-hydroxybenzaldehyde
ꢁ
ꢁ
ꢁ
ꢁ
N-(2,3,6,2 ,3 ,4 ,6 -hepta-O-acetyl-β-D-lactosyl)thiosemicarbazone (5r)
From 709 mg of 3 and 166 mg of 4r. Recrystallized from 96% ethanol to
yield white crystals of 5r. Yield: 659 mg, 77% (reaction time: 5 min, proce-
◦
25
D
dure A); 831 mg, 97% (reaction time: 2 min, procedure B); mp 140–142 C; [α]
–
1
+
104.3 (c 1.0, CHCl3); FTIR (KBr) ν / cm : 3539, 3277, 1757, 1734, 1604, 1371,
1
ꢁꢁꢁ
1
240, 1036; H NMR (DMSO-d6): δ (ppm) 11.80 (s, 1H, NH-2), 9.45 (s, 1H, 4 -
OH), 8.45 (d, 1H, J = 9.0 Hz, NH-4), 7.97 (s, 1H, CH N), 7.40 (d, 1H, J =
.5 Hz, H-2 ), 7.11 (dd, 1H, J = 8.5, 1.5 Hz, H-6 ), 6.82 (d, 1H, J = 8.5 Hz,
ꢁ
ꢁꢁ
ꢁꢁꢁ
1
ꢁ
ꢁꢁ
ꢁ
ꢁ
H-5 ), 5.75 (t, 1H, J = 9.0 Hz, H-1 ), 5.31 (t, 1H, J = 9.25 Hz, H-3 ), 5.24 (d,
ꢁ
ꢁ
ꢁ
1
H, J = 3.5 Hz, H-4 ), 5.15 (dd, 1H, J = 10.25, 3.75 Hz, H-2 ), 5.13 (t, 1H,
ꢁ
ꢁ
ꢁꢁ
J = 9.5 Hz, H-3 ), 4.87 (dd, 1H, J = 8.0, 10.5 Hz, H-2 ), 4.78 (d, 1H, J =
ꢁ
ꢁ
ꢁ
8
5
.0 Hz, H-1 ), 4.29 (d, 1H, J = 10.5 Hz, H-6 a), 4.25 (t, 1H, J = 6.5 Hz, H-
ꢁ
ꢁ
ꢁꢁꢁ
), 4.11 (q, 2H, J = 7.0 Hz, 3 -OCH2CH3), 4.08 (dd, 1H, J = 12.0, 5.5 Hz,

N-(Per-O-acetylated β-D-Lactosyl)Thiosemicarbazones
5
39
ꢁ
ꢁ
ꢁꢁ
ꢁꢁ
H-6 b), 4.04–4.01 (m, 2H, H-6 a & H-6 b), 3.86 (ddd, 1H, J = 10.0, 5.5, 1.5 Hz,
ꢁ
H-5 ), 3.81 (t, 1H, J = 9.25 Hz, H-4 ), 2.11–1.90 (s, 21H, 7×COCH3), 1.35 (t,
ꢁ
ꢁꢁ
13
3
1
1
4
(
1
H, J = 7.0 Hz, 3 -OCH2CH3); C NMR (DMSO-d6): δ (ppm) 177.8 (C S),
ꢁ
ꢁꢁ
ꢁꢁꢁ
70.2–169.0 (7×COCH3), 149.6 (C-4 ), 148.1 (C-3 ), 144.3 (CH N), 125.1 (C-
ꢁ
ꢁ
ꢁꢁ
ꢁꢁꢁ
ꢁꢁꢁ
ꢁꢁꢁ
ꢁꢁ
ꢁ
), 122.6 (C-6 ), 115.3 (C-5 ), 109.5 (C-2 ), 99.6 (C-1 ), 81.6 (C-1 ), 76.1 (C-
ꢁ
ꢁ
ꢁ
ꢁꢁ
ꢁꢁ
ꢁꢁ
), 73.3 (C-5 ), 72.4 (C-3 ), 70.8 (C-2 ), 70.3 (C-3 ), 68.8 (C-2 ), 69.7 (C-5 ), 67.1
ꢁ
ꢁ
ꢁꢁꢁ
ꢁ
ꢁꢁ
C-4 ), 63.9 (3 -OCH2CH3), 62.2 (C-6 ), 60.9 (C-6 ), 20.6–20.2 (7×COCH3),
ꢁ
ꢁꢁ
+
+
4.7 (3 -OCH2CH3); ESI-MS: m/z: 880.10 [M+Na] , 858.08 [M+H] , 803.08,
40.62, 512.74, 429.44, 413.40, 318.51; calc. for C36H47N3O19S = 857.25 Da.
5
Anal. calcd. for C36H47N3O19S: C, 50.40; H, 5.52; N, 4.90; S, 3.74. Found: C,
50.43; H, 5.50; N, 4.93; S, 3.72.
4
-Dimethylaminobenzaldehyde
ꢁ
ꢁ
ꢁ
ꢁ
N-(2,3,6,2 ,3 ,4 ,6 -hepta-O-acetyl-β-D-lactosyl)thiosemicarbazone (5s)
From 709 mg of 3 and 149 mg of 4s. Recrystallized from 96% ethanol to
yield white crystals of 5s. Yield: 655 mg, 78% (reaction time: 5 min, proce-
◦
25
D
dure A); 804 mg, 96% (reaction time: 2 min, procedure B); mp 187–188 C; [α]
–
1
+
112.5 (c 1.0, CHCl3); FTIR (KBr) ν / cm : 3320, 1753, 1604, 1372, 1220, 1054;
1
H NMR (DMSO-d6): δ (ppm) 11.67 (s, 1H, NH-2), 8.41 (d, 1H, J = 9.5 Hz, NH-
ꢁ
ꢁꢁ
ꢁꢁꢁ
4
2
), 7.97 (s, 1H, CH
N), 7.59 (d, 2H, J = 8.5 Hz, H-2 & H-6 ), 6.72 (d,
ꢁ
ꢁꢁ
ꢁꢁꢁ
ꢁ
H, J = 8.5 Hz, H-3 & H-5 ), 5.82 (t, 1H, J = 9.0 Hz, H-1 ), 5.30 (t, 1H,
ꢁ
ꢁꢁ
ꢁ
J = 9.0 Hz, H-3 ), 5.25 (d, 1H, J = 2.5 Hz, H-4 ), 5.17–5.14 (m, 2H, H-2 &
ꢁ
ꢁ
ꢁꢁ
ꢁꢁ
H-3 ), 4.88 (t, 1H, J = 9.0 Hz, H-2 ), 4.79 (d, 1H, J = 7.5 Hz, H-1 ), 4.30 (d,
ꢁ
ꢁꢁ
1
6
H, J = 11.0 Hz, H-6 a), 4.25 (t, 1H, J = 6.0 Hz, H-5 ), 4.09–4.05 (m, 1H, H-
ꢁ
ꢁꢁ
ꢁꢁ
ꢁ
b), 4.04–4.03 (m, 2H, H-6 a & H-6 b), 3.88–3.85 (m, 1H, H-5 ), 3.81 (t, 1H,
ꢁ
ꢁꢁꢁ
J = 9.25 Hz, H-4 ), 2.97 (s, 6H, 4 -N(CH3)2], 2.11–1.91 (s, 21H, 7×COCH3);
1
3
C NMR (DMSO-d6): δ (ppm) 177.3 (C
S), 170.2–169.0 (7×COCH3), 151.7
ꢁ
ꢁꢁ
ꢁꢁꢁ
ꢁꢁꢁ
ꢁꢁꢁ
ꢁꢁꢁ
(
C-4 ), 144.8 (CH
N), 128.9 (C-2 & C-6 ), 120.8 (C-1 ), 111.6 (C-3
&
ꢁ
ꢁꢁ
ꢁꢁ
ꢁ
ꢁ
ꢁ
ꢁ
ꢁ
C-5 ), 99.6 (C-1 ), 81.1 (C-1 ), 76.1 (C-4 ), 73.4 (C-5 ), 72.7 (C-3 ), 71.0 (C-2 ),
ꢁ
ꢁ
ꢁꢁ
ꢁꢁ
ꢁꢁ
ꢁ
ꢁꢁ
7
[
[
0.4 (C-3 ), 69.7 (C-5 ), 68.9 (C-2 ), 67.1 (C-4 ), 62.3 (C-6 ), 60.9 (C-6 ), 40.0
ꢁ
ꢁꢁ
+
4 -N(CH3)2], 20.6–20.2 (7×COCH3); ESI-MS: m/z: 863.39 [M+Na] , 841.45
+
M+H] , 821.57, 799.47, 577.88, 536.38, 433.16, 331.38; calc. for C36H48N4O17S
=
840.27 Da. Anal. calcd. for C36H48N4O17S: C, 51.42; H, 5.75; N, 6.66; S, 3.81.
Found: C, 51.46; H, 5.72; N, 6.64; S, 3.84.
Biological Activity
Antimicrobial activities
All the synthesized compounds 5a–s were screened for their in vitro
antibacterial and antifungal activities by the twofold serial dilution tech-
nique using the principles of Clinical and Laboratory Standards Institute.[
Antimicrobial activity in vitro against Staphylococcus aureus (ATCC 11632),
58]

40 N. Dinh Thanh et al.
5
Escherchia coli (ATCC 25922), Pseudomonas aeruginosa (ATCC 25923), Kleb-
siella pneumonia (ATCC 4352), Staphylococcus epidermidis (ATCC 12228),
Bacillus subtilis (ATCC 11774), Enterobacter aerogenes (ATCC 13048) were
investigated by minimum inhibitory concentration (MIC) using agar dilu-
tions technique using Mueller–Hinton agar medium. Test compounds were
dissolved in dimethyl sulfoxide (DMSO) and applied to plates with serially
diluted compounds to be tested. The inoculum was prepared using a 4–6-h
broth adjusted to a turbidity equivalent of an 0.5 McFarland standard, diluted
5
in broth media to give a final concentration of 5 × 10 CFU/mL in the test
tray; they were covered and placed in plastic bags to prevent evaporation.
◦
The plates were incubated at 35 C for 18–20 h. The MIC was defined as the
lowest concentration of compound giving complete inhibition of visible growth.
Sulphamethoxazole and trimethoprim were used as reference (positive con-
trol) for bacteria. Also as negative control, antimicrobial effects of the solvents
(DMSO) were determined. According to the values of the controls, the results
were evaluated. Minimal inhibitory concentrations for each compound were
investigated against standard bacterial strains. The MIC values for all test
compounds and reference drug are shown in Table 1.
The compounds 5a–s were also evaluated for their in vitro antifungal ac-
tivity against Aspergillus niger (439), Candida albicans (ATCC 7754), Fusar-
ium oxysporum (M42), Saccharomyces cerevisiae (SH20) using agar dilution
method with Saburoud’s dextrose agar (Hi-Media). Suspensions of each mi-
croorganisms were prepared to contain 10 CFU/mL and applied to agar plates
which have been serially diluted with compounds to be tested. The plates were
◦
[58]
incubated at 26 C during 48 h and MIC’s were determined.
The MIC values
for all test compounds and reference drug are shown in Table 2.
The calculated logP of all synthesized thiosemicarbazones have been ob-
tained by using ACD/LogP in Advanced Chemistry Development (ACD/Labs)
Software (in silico Prediction of Physicochemical, ADME, and Toxicity Proper-
ties, Release 12, 2015).
Statistical analysis
All data on antioxidant activities were the average of triplicate analyses
and one-way analysis of variance was performed by ANOVA procedures. Signif-
icant differences between means were determined by Duncan’s Multiple Range
tests and P values <0.05 were regarded as significant and p values <0.01 were
very significant.
SUPPLEMENTARY MATERIAL
Supplemental data for this article can be accessed on the publisher’s website.

N-(Per-O-acetylated β-D-Lactosyl)Thiosemicarbazones
5
41
FUNDING
Financial support for this work was provided by Vietnam’s National Foun-
dation for Science and Technology Development (NAFOSTED), code 104.01-
2013.26.
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