Synthesis of novel succinic acid derivatives as potential matrix metalloproteinases inhibitors and anticancer medicine

Article abstract of DOI:10.2174/157017810790796336

In this paper we have designed different methods, according to the differences in nucleophilicity of amino, to conjugate tartaric acid or malic acid with two kinds of useful intermediates respectively via amide bond, envisaging they are promising matrix metalloproteinase inhibitors or anticancer medicine.

Full text of DOI:10.2174/157017810790796336

Letters in Organic Chemistry, 2010, 7, 155-158
155
Synthesis of Novel Succinic Acid Derivatives as Potential Matrix
Metalloproteinases Inhibitors and Anticancer Medicine
Huiming Huanga,b, Dan Chenb, Shaohua Lib and Chengmei Liu*,a
aState Key Laboratory of Food Science and Technology, Nanchang University, 235, Nanjing East Road, Nanchang
330047, P.R., China
bDepartment of Pharmacy, Nanchang University, 461, Bayi Road, Nanchang 330006, P.R., China
Received November 17, 2009: Revised December 10, 2009: Accepted January 02, 2010
Abstract: In this paper we have designed different methods, according to the differences in nucleophilicity of amino, to
conjugate tartaric acid or malic acid with two kinds of useful intermediates respectively via amide bond, envisaging they
are promising matrix metalloproteinase inhibitors or anticancer medicine.
Keywords: Tartaric acid, malic acid, thiadiazole, acylation, nucleophilic substitution, matrix metalloproteinase inhibitors.
INTRODUCTION
RESULTS AND DISCUSSION
Matrix metalloproteinases (MMPs) are a family of
structurally related zinc-containing enzymes that play an
important role in the breakdown of connective tissue. The
balance between MMPs and its endogenous inhibitors –
tissue inhibitors of metalloproteinase (TMP) maintains the
stabilization of cells. The over expression of MMPs will
induce excessive degradation of extracellular matrix (ECM).
This is closely related with many inflammatory, malignant
and degenerative diseases such as cancer, osteoarthritis,
rheumatoid arthritis and so on [1-4]. Therefore, finding
effective matrix metalloproteinase inhibitors (MMPIs) to
cure these diseases is very important. Design and synthesis
of MMPIs has always been a hot research field of medicinal
chemistry since nineties of the last century. The requirement
for a molecule to be an effective inhibitor of the MMPIs is a
functional group (e.g., carboxylic acid, hydroxamic acid, and
sulfhydryl, etc.) capable of chelating the active-site zinc (II)
ion, that may be at least one functional group which provides
a hydrogen bond interaction with the enzyme backbone, and
one or more side chains which undergo effective van der
Waals interactions with the enzyme subsites [5]. Tartaric
acid and malic acids, however, are often used to chelate
many metal ions.
Firstly four kinds of intermediates, hydrazides, 2-amino-
1,3,4-thiadiazole derivatives, 2-acetyl-malic anhydride and
diacetyl-tartaric anhydride are prepared as described in the
literature [10-13]. According to the differences in
nucleophilicity of amino, we take different acylation
reagents, anhydride or acyl chloride. The hydrazides amino
are comparatively active, so we can take anhydride as
acylation reagent. But the activity of 2-amino-1,3,4-
thiadiazoles amino proved extremely weak (ꢀ
= 7.4). In
NH2
order to get high yield we transform tartaric acid from
anhydride into acyl chloride as acylation reagent. Final
deprotection of hydroxyl groups is achieved by a solution of
K2CO3 in methanol [14] (Scheme 1).
As for 2-acetyl-malic anhydride, though there is a
reactivity difference between the two carbonyl groups, there
will still be two kinds of compounds in the course of
transforming anhydride into mono-methyl ester as Scheme 2
shown. But the product will be mainly the former when 2-
acetyl-malic anhydride reacts with 2-amino-1,3,4-thiadiazole
derivatives. Because the stereospecific blockade of 2-amino-
1,3,4-thiadiazoles is more larger than methanol and the
carbonyl group near the hydroxyl group is more active than
the other, therefore we choose 2-acetyl-malic anhydride as
acylation reagent. Further as Davenport and Watson’s
procedure may be followed [15]. But it will give little
amount of product if integration is done with 2-amino-1,3,4-
thiadiazole derivatives.
2-Amino-1,3,4-thiadiazole derivatives are well known as
compounds of a wide range of anticancer activity, including
in vivo conditions [6]. They are also applied in antibacterial
activity, and used as MMPIs, pesticide and so on [7,8].
Hydrazides are widely applied in bioassays including matrix
metalloproteinase inhibitors [9]. Therefore, we design to
conjugate tartaric acid and malic acid respectively via amide
bond, expecting the designed new compounds to be good
MMPIs.
In addition, we identified the structure of compounds (6)
that the hydroxyl group is attached to C(3) by comparing the
1H NMR of compound (6a) and compound (5a) (Table 1).
This result accords with our prediction.
CONCLUSION
In conclusion, several proposals to synthesize asymmetry
compounds and regional selectivity in chemical reactions are
described in this paper. Two kinds of scaffolds which have
*Address correspondence to this author at the State Key Laboratory of Food
Science and Technology, Nanchang University, 235, Nanjing East Road,
Nanchang 330047, P.R., China; Tel: +86-13803521460;
E-mail: cdan99199@yahoo.com.cn
1570-1786/10 $55.00+.00
© 2010 Bentham Science Publishers Ltd.

156 Letters in Organic Chemistry, 2010, Vol. 7, No. 2
Huang et al.
O
O
OH
O
AcO
H
OH
b, c
S
N
a
O
HO
R3
OH
OH
O
N
O
N
O
(1)
(6)
R3 = phenyl, p-telyl, 4-bromo-phenyl, 3-heptyl
O
O
OH
OAc
O
AcO
OH
O
e, f
d
O
HO
Cl
AcO
OH
O
O
OAc
O
(3)
(2)
g, h
i, j
OH
O
O
OH
O
H
H
N
S
N
R2
OH
R1
N
OH
H
N
O
OH
N
O
OH
(5)
(4)
R1 = phenyl, benzyl, p-fluoro-phenyl, p-methoxyl-phenyl R2 = phenyl, benzyl, 4-methoxy-phenyl, 3-heptyl
Scheme 1. (a) Acetyl chloride, 50 ˚C, 2 h, 90%; (b) 2-amino-1,3,4-thiadiazoles, THF, 0 ˚C, 48 h; (c) 1 M NaOH, 4 h; (d) acetic anhydride,
sulfuric acid, 120 ˚C, 10 min, 78% (e) dry MeOH, 0 ˚C, 15 min, 100%; (f) SOCl2, 75 ˚C, 2 h, 97%; (g) hydrazides, THF, 30 min; (h) 1 M
NaOH, 4 h; (i) 2-amino-1,3,4-thiadiazoles, triethylamine, THF, 24 h; (j) K2CO3, MeOH, H2O, 65 ˚C, 12 h.
O
O
O
O
AcO
O
OH
OH
HO
O
HO
O
OAc
O
OH
O
OAc
O
O
a
O
N
O
N
O
OH
H
N
b, c
OH
S
O
+
N
S
HO
H
O
O
OH
O
N
OH
O
N
Scheme 2. (a) 2,2-dimethoxypropane, tosic acid, 100%; (b) 2-amino-1,3,4-thiadiazole, DCC, THF; (c) THF, 2 M HCl.
Table 1. Structure Identification by Comparing the 1H NMR of Compound (6a) and (5a)
ꢀ (ppm)
-NH
ꢀ (ppm)
3-OH
ꢀ (ppm)
3-CH
ꢀ (ppm)
2-OH
ꢀ (ppm)
2-CH
ꢀ (ppm)
-COOH
Compound
OH
O
H
S
N
OH
12.18
12.40
5.89
6.00
4.63
4.60
5.37
4.47
12.78
12.40
N
O
OH
N
5a
OH
O
H
N
S
OH
2.61
2.76
\
N
O
N
6a

Synthesis of Novel Succinic Acid Derivatives
Letters in Organic Chemistry, 2010, Vol. 7, No. 2
157
extensive biological activity have been successfully
conjugated with tartaric acid and malic acid. We expect these
new compounds have better biological activity.
1578, 1528, 1254, 1123, 1072, 1026, 845; ESI-MS m/z:
[M+Na]+ 321.12; 1H NMR (DMSO-d6, 600 MHz): ꢀ 3.82 (s,
3H), 4.39 (s, 2H), 5.05 (s, 1H), 5.74 (s, 1H), 7.01 (d, J = 9.0
Hz, 2H), 7.88 (d, J = 9.0 Hz, 2H), 9.67 (s, 1H), 10.30 (s,
1H), 12.66 (s, 1H).
EXPERIMENTAL
All reagents were obtained from commercial suppliers
and used without further purification except SOCl2,
methanol and triethylamine. All the solvents were purified
before use. All reactions were monitored by thin-layer
chromatography on 0.25 mm silica gel plates (GF-254).
Melting points were determined on an electrothermal
melting point apparatus and were uncorrected. ESI mass
spectra were recorded using a Waters ZQ4000/2695 LC–MS
spectrometer. Infrared spectra were obtained as KBr pellets
Synthesis of 2,3-dihydroxy-N-(5-phenyl- [1,3,4]thiadiazol-
2-yl)-succinamic acid (5a)
To the solution of 2-amino-5-phenyl-1,3,4- thiadiazole
(3.54 g, 0.020 mol) in THF and excess triethylamine
dropwise added the solution of compound (3) (6.40 g, 0.024
mol) in THF which was derived from diacetyl-tartaric
anhydride reacted with anhydrous methanol and then SOCl2.
The solvent was removed with a rotary evaporator after 24
hours and the residue was hydrolyzed in a solution of K2CO3
in methanol at 65°C for 12 hours to afford compound (5a)
(5.31 g, 86%); Mp 210°C (decomposition); IR (KBr, cmꢁ1):
3242, 1736, 1693, 1533, 1466, 1304, 1269, 1146, 1082, 762,
683; ESI-MS m/z: [M+H]+ 310.08; 1H NMR (DMSO-d6, 600
MHz): ꢀ 4.47 (d, J = 1.8 Hz, 1H), 4.63 (s, 1H), 5.37 (s, 1H),
5.89 (s, 1H), 7.53-7.55 (m, 3H), 7.95-7.96 (q, 2H), 12.18 (s,
1H), 12.78 (s, 1H).
1
on a Shimadzu FTIR-8000 spectrometer. H NMR spectra
were obtained in DMSO on a Bruker 600 MHz spectrometer.
The chemical shifts were reported in ꢀ values (ppm) relative
to tetramethylsilane (TMS) as internal standard.
Synthesis of (2R, 3R)-2,3-dihydroxy-4-(N'-Benzoyl-hydr-
azino)-4-oxo-butyric acid (4a)
The product (4a) was prepared from the reaction of
diacetyl-l-tartaric anhydride with benzoic acid hydrazide in
THF for 30 minutes. Finally, acetyls were removed by 1 M
NaOH. Purification was by recrystallization from chloroform
if necessary. White solid (yield 92%); Mp 128-130°C; IR
(KBr, cmꢁ1): 3533, 3476, 3360, 3213, 3028, 1744, 1693,
1651,1578, 1528, 1273, 1123, 1072, 694; ESI-MS m/z:
[M+Na]+ 291.10; 1H NMR (DMSO-d6, 600 MHz): ꢀ 4.39 (s,
2H), 5.04 (s, 1H), 5.76 (d, J = 6.0 Hz, 1H), 7.47-7.50 (q,
2H), 7.55-7.58 (m, 1H), 7.89-7.90 (q, 2H), 9.74 (d, J = 1.2
Hz, 1H), 10.44 (d, J = 1.2 Hz, 1H), 12.68 (s, 1H).
2,3-dihydroxy-N-(5-Benzyl-[1,3,4]thiadiazol-2-yl)-succ-
inamic acid (5b)
White solid (4.39 g, 68%); Mp 162-163°C; IR (KBr,
cmꢁ1): 3256, 1736, 1693, 1533, 1308, 1269, 1150, 1082, 700,
669; ESI-MS m/z: [M+H]+ 324.02; 1H NMR (DMSO-d6, 600
MHz): ꢀ 4.36 (s, 2H), 4.40 (d, J = 1.8 Hz, 1H), 4.55 (s, 1H),
5.30 (s, 1H), 5.83 (s, 1H), 7.26-7.29 (m, 1H), 7.32-7.36 (m,
4H), 11.89 (s, 1H), 12.74 (s, 1H).
2,3-dihydroxy-N-[5-(4-methoxy-phenyl)-[1,3,4]thiadiazol-
2-yl]-succinamic acid (5c)
(2R, 3R)-2,3-dihydroxy-4-(N'-phenylacetyl- hydrazino)-4-
oxo-butyric acid (4b)
White solid (6.03 g, 89%); Mp 193-194°C; IR (KBr,
cmꢁ1): 3240, 1740, 1693, 1609, 1535, 1462, 1308, 1258,
White solid (yield 81%); Mp 120-122°C; IR (KBr, cmꢁ1):
3530, 3476, 3387, 3206, 3028, 1740, 1697, 1659, 1570,
1520, 1420, 1238, 1111, 1072, 721, 694; ESI-MS m/z:
[M+Na]+ 305.13; 1H NMR (DMSO-d6, 600 MHz): ꢀ 3.48 (s,
2H), 4.33 (s, 2H), 5.02 (s, 1H), 5.71 (s, 1H), 7.23 (t, J = 3.0
Hz, 1H), 7.28-7.30 (q, 4H), 9.62 (d, J = 2.4 Hz, 1H), 10.25
(d, J = 2.4 Hz, 1H), 12.64 (s, 1H).
1
1142, 1084, 1034, 833; ESI-MS m/z: [M+H]+ 340.07; H
NMR (DMSO-d6, 600 MHz): ꢀ 3.83 (s, 3H), 4.47 (s, 1H),
4.61 (d, J = 3.6 Hz, 1H), 5.36 (s, 1H), 5.88 (d, J= 6.0 Hz,
1H), 7.09 (dd, J= 1.8 Hz, 7.2 Hz, 2H), 7.89 (dd, J = 1.8 Hz,
6.6 Hz, 2H), 12.07(s, 1H), 12.78 (s, 1H).
2,3-dihydroxy-N-[5-(1-Ethyl-pentyl)-[1,3,4]thiadiazol-2-
yl]-succinamic acid (5d)
(2R, 3R)-2,3-dihydroxy-4-[N'-(4-Fluoro-benzoyl)-hydra-
zino]-4-oxo-butyric acid (4c)
White solid (4.30 g, 65%); Mp 130-132°C; IR (KBr,
cmꢁ1): 3233, 2959, 2932, 2858, 1697, 1535, 1458, 1312,
1142, 1080; ESI-MS m/z: [M+H]+ 332.09; 1H NMR
(DMSO-d6, 600 MHz): ꢀ 0.80-0.84 (m, 6H), 1.20-1.30 (m,
4H), 1.61-1.65 (m, 2H), 1.70-1.75 (m, 2H), 2.99-3.04 (m,
1H), 4.43 (d, J = 2.4 Hz, 1H), 4.57 (d, J = 2.4 Hz, 1H), 5.34
(s, 1H), 5.84 (s, 1H), 11.86 (s, 1H), 12.76 (s, 1H).
White solid (yield 84%); Mp 165-166°C; IR (KBr, cmꢁ1):
3533, 3479, 3340, 3209, 3017, 1740, 1693, 1651, 1605,
1508, 1273, 1246, 1123, 1072, 853; ESI-MS m/z: [M+Na]+
309.10; 1H NMR (DMSO-d6, 600 MHz): ꢀ 4.39 (s, 2H), 5.04
(s, 1H), 5.77 (d, J = 6.0 Hz, 1H), 7.33 (t, J = 9.0 Hz, 2H),
7.97 (dd, J = 5.4 Hz, 9.0 Hz, 2H), 9.75 (d, J = 1.8 Hz, 1H),
10.48 (d, J = 1.8 Hz, 1H), 12.69 (s, 1H).
Synthesis of 3-hydroxy-4-oxo-4-(5-phenyl-1,3,4-thiadia-
zol-2-ylamino)butanoic acid (6a)
(2R, 3R)-2,3-dihydroxy-4-[N'-(4-methoxy-benzoyl)-hydr-
azino]-4-oxo-butyric acid (4d)
White solid (yield 87%); Mp 121-122°C; IR (KBr, cmꢁ1):
3533, 3479, 3340, 3206, 3016, 1740, 1690, 1647, 1608,
The product was prepared from the reaction of compound
(1) (3.79 g, 0.024 mol) with 2-amino-5-phenyl-1,3,4-
thiadiazole (3.54 g, 0.020 mol) in THF at 0°C for 48 hours.

158 Letters in Organic Chemistry, 2010, Vol. 7, No. 2
Huang et al.
Purification is by recrystallization from water and acetic
acid. Yield 52%; Mp 192-193°C; IR (KBr, cmꢀ1): 3317,
1701, 1543, 1462, 1439, 1308, 1211, 1177, 1107, 764, 687;
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ESI-MS m/z: [M+H]+ 294.08; H NMR (DMSO-d6, 600
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1103, 837; ESI-MS m/z: [M+2]+ 374.00; H NMR (DMSO-
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1308, 1277, 1215, 1099; ESI-MS m/z: [M+H]+ 316.24; H
NMR (DMSO-d6, 600 MHz): ꢁ 0.80-0.84 (m, 6H), 1.13-1.30
(m, 4H), 1.61-1.64 (m, 2H), 1.71-1.75 (q, 2H), 2.57 (dd, J =
7.2 Hz, 15.6 Hz, 1H), 2.73 (dd, J = 5.4 Hz, 15.6 Hz, 1H),
3.00-3.03 (m, 1H), 4.55 (dd, J = 5.4 Hz, 7.2 Hz, 1H), 5.93 (s,
1H), 12.14 (s, 1H), 12.30 (s, 1H).
ACKNOWLEDGEMENT
Thank Natural Science Foundation of JiangXi province
for financial support.