Synthesis using microwave irradiation, characterisation and antibacterial activity of Novel deoxycholic acid-triazole conjugates

Article abstract of DOI:10.3184/174751912X13364636691591

Novel deoxycholic acid 3α-triazole conjugates based on methyl 3α-chloroacetoxy-12α-hydroxy-cholanate have been synthesised. The synthesis is accelerated by microwave irradiation under solvent free conditions in the presence of K₂CO₃. Some of these compounds were tested for antibacterial activity against B.subtilis, P.aeruginosa and S.aureus. The preliminary results indicated that these deoxycholic acid-triazole conjugates have good inhibitory effect against B.subtilis. All of the compounds were characterised by ¹H NMR, IR, ESI-MS spectra and elemental analyses.

Full text of DOI:10.3184/174751912X13364636691591

JOURNAL OF CHEMICAL RESEARCH 2012
RESEARCH PAPER 383
JULY, 383–386
Synthesis using microwave irradiation, characterisation and antibacterial
activity of novel deoxycholic acid-triazole conjugates
Jie Yang, Zhigang Zhao* and Hui Li
College of Chemistry and Environmental Protection Engineering, Southwest University for Nationalities, Chengdu 610041, P. R. China
Novel deoxycholic acid 3α-triazole conjugates based on methyl 3α-chloroacetoxy-12α-hydroxy-cholanate have been
synthesised. The synthesis is accelerated by microwave irradiation under solvent free conditions in the presence of
K2CO3. Some of these compounds were tested for antibacterial activity against B.subtilis, P.aeruginosa and S.aureus.
The preliminary results indicated that these deoxycholic acid-triazole conjugates have good inhibitory effect against
B.subtilis. All of the compounds were characterised by 1H NMR, IR, ESI-MS spectra and elemental analyses.
Keywords: deoxycholic acid, triazole, microwave irradiation, antibacterial activity
Bile acids are pharmacologically interesting as potential
carriers of liver-specific drugs, absorption enhancers, and
cholesterol lowering agents.1 Moreover, owing to their own
facially amphiphilic structures, it is possible to utilise bile
acids as building blocks for other more complex amphiphilic
molecules.2 A variety of bile acids derivatives with anti-tumour,
anti-microbial, anti-HCV, anti-HIV, carbonic anhydrase
inhibitors and glucocorticoid receptor antagonists have been
reported.3–8 However, the common feature of these compounds
are that the side chains of the bile acids are changed, whereas
the hydroxyl groups are preserved. To the best of our knowl-
edge, little work has been published on the synthesis and phys-
icochemical properties of compounds in which the hydroxyl
groups are replaced by heterocyclic compounds containing
hydrophilic groups.
diverse biological activities such as anti-bacterial, anti-prolif-
erative, anti-cancer, analgesic and anti-oxidant agents.10–14
In recent years, microwave-assisted organic synthesis has
been applied to a large and expanding range of chemical trans-
formations, and this method of energy transfer to a reaction
mixture can provide impressive enhancements in product
yield, selectivity, and reaction rate.15,16 In view of this, we now
report a simple, efficient and rapid method for the synthesis
of deoxycholic acid-triazole conjugates under microwave
irradiation. The synthetic route is depicted in Scheme 1.
Result and discussion
In the 1H NMR (DMSO-d6, 400 MHz) spectrum of compounds
7a–j the protons of the ArH appeared at 8.12–7.06 ppm.
A single peak in the region 6.61–5.72 ppm attributed to
the NH2 group. The 3β-H and 12β-H appeared in the region
4.70–4.64 ppm (multiplet) and 4.28–4.24 ppm, respectively.
The 12α-OH appeared at 3.78 ppm (singlet). The resonance
Azoles are the largest class of antibacterial agents in clinical
use.9 1,2,4-Triazole moieties are attractive connecting units,
and synthetic molecules containing 1,2,4-triazole units show
Scheme 1 Reagents and conditions: (i) SOCl2, C2H5OH, MWI; (ii) NH2NH2·H2O, MWI; (iii) CS2, KOH, C2H5OH; (iv) NH2NH2·H2O, MWI;
(v) CH3COCl, CH3OH; (vi) pyridine, ClCH2COCl, CHCl3, MWI; (vii) 4a–j, K2CO3, DMF, MWI.
* Correspondent. E-mail: zzg63129@yahoo.com.cn

384 JOURNAL OF CHEMICAL RESEARCH 2012
corresponding to –OCOCH2N– protons was identified at the
region 4.14–4.04 ppm (singlet). Moreover, the three hydrogen
singlets in the range 0.92–0.90, 0.87–0.85 and 0.60–0.59 ppm
were assigned to the 21-CH3, 19-CH3 and 18-CH3, respec-
tively. The IR spectra of all compounds 7a–j revealed strong
characteristic bands in the region 3361–3310 cm−1, 3286–
3170 cm−1 and 1732–1729 cm−1 which were assigned to the
12α-OH NH2 and –COOCH3, respectively. The strong bands at
1631–1572 cm−1 corresponded with the absorption of C=N.
The most characteristic peaks were the bands at 1384–1379
cm−1 and 1094–1040 cm−1 assigned to the C=S. In addition,
they gave satisfactory elemental analyses and mass spectra.
Compounds 7a–j showed molecular ion peak at [2M+H]+ (or
[2M+Na]+).
count, compounds were dissolved in DMSO. In each case,
0.1 mL of test compound and 0.1 mL test bacterial suspension
were pipette to provide a uniform coating on the tablet. The
bacteria tablets were incubated at 37 °C for 24 h. The results
are presented in Table 4, Figs 1,2 and 3. The preliminary results
indicated that these deoxycholic acid-triazole conjugates
have good inhibitory effects against B.subtilis. The details of
compounds 7a–j for their antibacterial activity are under
further studies.
Experimental
Melting points were determined on a micro-melting point apparatus
and the thermometer was uncorrected. IR spectra were obtained on
1
1700 Perkin-Elmer FTIR using KBr disks. H NMR spectra were
In conclusion, all of the spectroscopy and elemental
analysis data were consistent with the structures of the target
compounds.
Comparison of microwave irradiation and conventional
heating: In order to identify the best reaction conditions,
we examined the synthesis of 7a with different microwave
irradiation power and reaction time.
As shown in Table 1, the reaction was irradiated with differ-
ent power for the same reaction time (20 min). As a result,
150 W was the optimum power and the use of decreased the
yield.
As shown in Table 2, the reaction was irradiated for a differ-
ent time with the same power (150 W). It was found that
20 min was the best reaction time.
As shown in Table 3, compared to conventional method,
microwave method decreased the reaction time from 1560–
2880 min to 20–50 min. It was obvious that the yields increased
from 45–55% to 93–98%. From these data, we conclude that
microwave irradiation method is a simple, efficient and rapid
synthetic method.
In vitro antibacterial activity: Compounds 7c, 7d, 7g, 7j
were evaluated for their antibacterial activities by the method
of plate culture count. The Gram-positive and Gram-negative
bacteria which utilised in this study consisted of B.subtilis,
P.aeruginosa and S.aureus. In the method of plate culture
recorded on a Varian INOVA 400 MHz spectrometer using TMS as
internal standard. Mass spectra were determined on Finnigan LCQDECA
instrument. Elemental analysis was performed on a Carlo-Erba-1106
autoanalyser. All reactions were performed in a commercial micro-
wave reactor (XH-100A, 100–1000 W, Beijing Xianghu Science
and Technology Development Co. Ltd, Beijing, P.R. China). Optical
rotations were measured on a Wzz-2B polarimeter. All the solvents
were purified before use.
Synthesis of compounds 4a–j;17 typical procedure
Substituted acid was esterified with ethanol in the presence of SOCl2.
Then the ester refluxed with hydrazine hydrate to obtain carbohydra-
zide. The carbohydrazide on reaction with carbon disulphide in
ethanolic potassium hydroxide yielded the corresponding dithiocarba-
zinate in good yield then used directly for the next step without further
purification. Triazole was synthesised by refluxing dithiocarbazinate
with hydrazine hydrate. The crude product was then recrystallised
from ethanol to give a pure sample. (Table 5)
Synthesis of compound 5;18 general procedure
Deoxycholic acid (5 mmol) and anhydrous methanol (50 mL) were
added to a round-bottomed flask and the mixture was stirred in ice
bath. Then pure acetyl chloride was added slowly. The reaction was
monitored by TLC. The reaction solution was then poured into ice
water, and the required solid was precipitated out. It was filtered, dried
under vacuum. The crude product was purified by column chromato-
graphy on silica gel using V(CHCl3): V(CH3COCH3): V(CH3OH) =
70:20:1 as elution agent to give 1.82 g. White solid, yield 90%, m.p.
79–80 ºC (lit.18 m.p. 74–76 ºC).
Synthesis of compound 6;19 general procedure
Table 1 Effect of microwave powers on yields
Compound 5 (1 mmol) and chloroacetyl chloride (1.2 mmol) were
dissolved in dry chloroform (20 mL).A solution of anhydrous pyridine
(1mmol) in dry chloroform (10 mL) was added slowly. Then, the
mixture was refluxed in the microwave reactor (TLC). The mixture
was evaporated to dryness under vacuum, and the residue was
dissolved in CH3COOC2H5, The solution was washed with NaHCO3,
brine, dried over Na2SO4 and evaporated to dryness under vacuum.
The impure product was then purified by flash chromatography
(elution with CH3COOC2H5: petroleum ether = 1:5) to give 0.44g.
White solid, yield 92%; m.p. 130–132 ºC (lit.19 134–135 ºC).
Power/W
Yields/%
100
–
125
60
150
94
175
85
200
0
Table 2 Effect of microwave irradiation time yields
Time/min
Yields/%
10
80
20
95
30
90
40
88
50
79
Synthesis of compounds 7a–j; microwave method
Triazole(0.5mmol), methyl3α-chloroacetoxy-12α-hydroxy-cholanoate
(0.5 mmol) and anhydrous K2CO3 (1.0 g) were placed in a porcelain
mortar. After grinding, dry DMF (the amount of catalyst) was added
and the mixture was placed in the microwave reactor. Then it was
irradiated for 20–50 min at 150 W (TLC). The mixture was cooled to
room temperature and dissolved in CH3COOC2H5. It was filtered
and evaporated to dryness under vacuum. The impure product was
then purified by flash chromatography (elution with CH3COOC2H5:
petroleum ether = 4:1–8:1) to give 7a–j (92–98%).
Methyl 3α-(2-(4-amino-3-phenyl-5-thioxo-4,5-dihydro-1,2,4-triazol-
1-yl)acetoxy)-12α-hydroxy-cholanate (7a): White solid, yield 95%,
m.p. 91–93 ºC; [α]2D0–71.7 (c 0.19, CH2Cl2); IR (cm−1): 3339, 3191,
2939, 2867,1730, 1631, 1456, 1379, 1296, 1172, 1042, 772, 695; 1H
NMR (DMSO-d6, 400 MHz): δ 7.96 (d, J = 2.4 Hz, 2H, ArH), 7.51 (s,
3H, ArH), 6.21 (s, 2H, NH2), 4.67 (bs, 1H, 3β-H), 4.24 (s, 1H, 12β-H),
4.07 (s, 2H, –OCOCH2N), 3.78 (s, 1H, 12α-OH), 3.57 (s, 3H,
–COOCH3), 0.91 (d, J = 6.4 Hz, 3H, 21-CH3), 0.87 (s, 3H, 19-CH3),
Table 3 Comparison of the synthesis of 7a–j using microwave
and conventional methods
a
Compd Conventional method Microwave method tc/tw
Time/min Yields/% Time/min Yield%
7a
7b
7c
7d
7e
7f
1440
2040
2220
2520
2640
2880
2520
2760
2520
1800
50
45
49
55
50
51
46
49
45
55
20
30
25
30
45
50
25
40
45
35
95
93
97
95
93
95
98
95
92
96
72
68
88
84
58
57
100
69
56
51
7g
7h
7i
7j
tc, conventional method time; tw, microwave method time

JOURNAL OF CHEMICAL RESEARCH 2012 385
Table 4 The antibacterial rate of compounds 7c, 7d, 7g and 7j against B. subtilis, P. aeruginosa and S. aureus (%)
512 µg mL−1
256 µg mL−1
128 µg mL−1
64 µg mL−1
32 µg mL−1
7c
B.subtilis
P.aeruginosa
S.aureus
B.subtilis
P.aeruginosa
S.aureus
B.subtilis
P.aeruginosa
S.aureus
B.subtilis
P.aeruginosa
S.aureus
B.subtilis
P.aeruginosa
S.aureus
89.6
21.5
50.8
83.1
34.7
22.6
66.2
7.9
88.3
26.7
4.8
77.9
16.9
16.1
61.0
31.4
–
70.1
–
–
100.0
100.0
95.5
59.7
6.1
49.3
28.6
–
53.2
18.3
–
6.4
20.1
–
44.1
19.2
–
100.0
100.0
75.5
44.1
23.9
–
36.3
31.4
–
–
2.3
–
–
22.5
–
100.0
100.0
37.7
–
7d
72.7
31.9
–
32.4
23.4
–
64.9
22.0
–
100.0
100.0
88.8
7g
–
7j
74.0
35.6
–
100.0
100.0
100.0
Amoxicillin
–No antibacterial activity.
Table 5 The melting point of triazole 4a–j
Compd
Formula
M.p./ °C
Lit. m.p./ °C
4a
4b
4c
4d
4e
4f
C8H8N4S
C9H10N4S
C9H10N4S
202–204
203–205
201–203
211–212
206–207
149–150
213–214
214–216
208–210
204–205
203–20420
206–20821
20121
C9H N4OS
216–21921
20821
10
C9H N4OS
10
C10H12N4OS
C8H7N4ClS
C8H7N4ClS
C8H7N4BrS
C12H10N4S
153–15422
215–21721
210–21221
205–20621
20623
4g
4h
4i
4j
Fig. 1 Comparison of compounds 7c, 7d, 7g, and 7j against
B.subtilis antibacterial rate.
3340, 3187, 2937, 2867, 1731, 1613, 1452, 1382, 1296, 1185, 1094,
978, 793, 714; 1H NMR (DMSO-d6, 400 MHz): δ 7.77 (t, J =
6.4 Hz, 2H, ArH), 7.40 (t, J = 7.2 Hz, 1H, ArH), 7.31 (d, J = 7.6 Hz,
1H, ArH), 6.19 (s, 2H, NH2), 4.67–4.64 (m, 1H, 3β-H), 4.27 (s, 1H,
12β-H), 4.06 (s, 2H, –OCOCH2N), 3.78 (s, 1H, 12α-OH), 3.57 (s, 3H,
–COOCH3), 2.38 (s, 3H, Ar-CH3), 0.91 (d, J = 6.0 Hz, 3H, 21-CH3),
0.87 (s, 3H, 19-CH3), 0.59 (s, 3H, 18-CH3). ESI-MS m/z (%): 1305
([2M+H]+, 100). Anal. Calcd for C36H52N4O5S: C, 66.23; H, 8.03; N,
8.58. Found: C, 65.93; H, 8.07; N, 8.55%.
Methyl 3α-(2-(4-amino-3-(4-methylphenyl)-5-thioxo-4,5-dihydro-
1,2,4-triazol-1-yl)-acetoxy)-12α-hydroxy-cholanate (7c): White solid,
yield 97%, m.p. 104–106 ºC; [α]2D0–80.2 (c 0.17, CH2Cl2); IR (cm−1):
3343, 3195, 2939, 2868, 1730, 1623, 1452, 1380, 1297, 1173, 1045,
983, 823, 726; 1H NMR (DMSO-d6, 400 MHz): δ 7.86 (d, J = 8.0 Hz,
2H, ArH), 7.32 (d, J = 8.0 Hz, 2H, ArH), 6.17 (s, 2H, NH2), 4.66–4.64
(m, 1H, 3β-H), 4.26 (s, 1H, 12β-H), 4.05 (s, 2H, –OCOCH2N), 3.77 (s,
1H, 12α-OH), 3.57 (s, 3H, –COOCH3), 2.37 (s, 3H, Ar-CH3), 0.90 (d,
J = 6.0 Hz, 3H, 21-CH3), 0.86 (s, 3H, 19-CH3), 0.59 (s, 3H, 18-CH3);
ESI-MS m/z (%): 1305 ([2M+H]+, 100). Anal. Calcd for C36H52N4O5S:
C, 66.23; H, 8.03; N, 8.58. Found: C, 66.36; H, 8.06; N, 8.61%.
Methyl3α-(2-(4-amino-3-(2-methoxyphenyl)-5-thioxo-4,5-dihydro-
1,2,4-triazol-1-yl)-acetoxy)-12α-hydroxy-cholanate (7d): White solid,
yield 95%, m.p. 102–104 ºC; [α]2D0 –139.8 (c 0.13, CH2Cl2); IR (cm−1):
3361, 3286, 2939, 2867, 1732, 1610, 1529, 1471, 1379, 1251, 1169,
1044, 985, 758; 1H NMR (DMSO-d6, 400 MHz): δ 7.54 (t, J = 8.0 Hz,
1H, ArH), 7.41 (d, J = 7.2 Hz, 1H, ArH), 7.19 (d, J = 8.4 Hz, 1H,
ArH), 7.08 (t, J = 7.2 Hz, 1H, ArH), 5.72 (s, 2H, NH2), 4.70–4.64 (m,
1H, 3β-H), 4.28 (s, 1H, 12β-H), 4.06 (s, 2H, –OCOCH2N), 3.82
(s, 3H, Ar–OCH3), 3.77 (s, 1H, 12α-OH), 3.57 (s, 3H, –COOCH3),
0.91 (d, J = 6.0 Hz, 3H, 21-CH3), 0.87 (s, 3H, 19-CH3), 0.59 (s, 3H,
18-CH3); ESI-MS m/z (%): 1337 ([2M+H]+, 100). Anal. Calcd for
C36H52N4O6S: C, 64.64; H, 7.84; N, 8.38. Found: C, 64.49; H, 7.88; N,
8.35%.
Fig. 2 Comparison of compounds 7c, 7d, 7g and 7j against
P.aeruginosa antibacterial rate.
Fig. 3 Comparison of compounds 7c, 7d, 7g and 7j against
S.aureus antibacterial rate.
0.59 (s, 3H, 18-CH3); ESI-MS m/z (%): 1277 ([2M+H]+, 100). Anal.
Calcd for C35H50N4O5S: C, 65.80; H, 7.89; N, 8.77. Found: C, 66.06;
H, 7.86; N, 8.80%.
Methyl 3α-(2-(4-amino-3-(3-methylphenyl)-5-thioxo-4,5-dihydro-
1,2,4-triazol-1-yl)-acetoxy)-12α-hydroxy-cholanate (7b): White solid,
yield 93%, m.p. 171–173 ºC; [α]2D0 –129.8 (c 0.14, CH2Cl2); IR (cm−1):
Methyl3α-(2-(4-amino-3-(4-methoxyphenyl)-5-thioxo-4,5-dihydro-
1,2,4-triazol-1-yl)-acetoxy)-12α-hydroxy-cholanate (7e): White solid,
yield 93%, m.p. 175–177 ºC; [α]2D0 –53.4 (c 0.17, CH2Cl2); IR (cm−1):
3310, 3170, 2940, 2868, 1732, 1611, 1464, 1381, 1299, 1177, 1033,
986, 832; 1H NMR (DMSO-d6, 400 MHz): δ 7.91 (d, J = 8.4 Hz, 2H,
ArH), 7.06 (d, J = 8.8 Hz, 2H, ArH), 6.61 (s, 2H, NH2), 4.69–4.63 (m,

386 JOURNAL OF CHEMICAL RESEARCH 2012
1H, 3β-H), 4.26 (s, 1H, 12β-H), 4.04 (s, 2H, –OCOCH2N), 3.81
(s, 3H, Ar–OCH3), 3.78 (s, 1H, 12α-OH), 3.57 (s, 3H, –COOCH3),
0.90 (d, J = 6.0 Hz, 3H, 21-CH3), 0.86 (s, 3H, 19-CH3), 0.59 (s, 3H,
18-CH3); ESI-MS m/z (%): 1337 ([2M+H]+, 100). Anal. Calcd for
C36H52N4O6S: C, 64.64; H, 7.84; N, 8.38. Found: C, 64.85; H, 7.80;
N, 8.40%.
Methyl 3α-(2-(4-amino-3-(2-ethoxyphenyl)-5-thioxo-4,5-dihydro-
1,2,4-triazol-1-yl)-acetoxy)-12α-hydroxy-cholanate (7f): White solid,
yield 95%, m.p. 101–103 ºC; [α]2D0 –85.2 (c 0.16, CH2Cl2); IR (cm−1):
3358, 3286, 2938, 2867, 1733, 1609, 1466, 1380, 1247, 1167, 1040,
986, 756; 1H NMR (DMSO-d6, 400 MHz): δ 7.52 (t, J = 7.4 Hz, 1H,
ArH), 7.43 (d, J = 7.2 Hz, 1H, ArH), 7.18 (d, J = 8.4 Hz, 1H, ArH),
7.08 (t, J = 7.4 Hz, 1H, ArH), 5.72 (s, 2H, NH2), 4.69–4.64 (m, 1H,
3β-H), 4.28 (s, 1H, 12β-H), 4.19–4.10 (m, 2H, Ar–OCH2CH3), 4.06
(s, 2H, –OCOCH2N), 3.78 (s, 1H, 12α-OH), 3.57 (s, 3H, –COOCH3),
1.28 (t, J = 7.0 Hz, 3H, Ar–OCH2CH3), 0.91 (d, J = 6.0 Hz, 3H,
21-CH3), 0.87 (s, 3H, 19-CH3), 0.59 (s, 3H, 18-CH3); ESI-MS m/z
(%): 1365 ([2M+H]+, 100). Anal. Calcd for C37H54N4O6S: C, 65.07; H,
7.97; N, 8.20. Found: C, 64.74; H, 8.01; N, 8.23%.
ESI-MS m/z (%): 1377 ([2M+H]+, 100). Anal. Calcd for C39H52N4O5S:
C, 67.99; H, 7.61; N, 8.13. Found: C, 67.65; H, 7.63; N, 8.17%.
Synthesis of compounds 7a–j; general procedure
In a typical procedure, the triazole (0.5 mmol) and anhydrous K2CO3
(0.5 mmol) in dry DMF (5 mL) was stirred at room temperature
for 2 h. Next, methyl 3α-chloroacetoxy-12α-hydroxy-cholanoate
(0.5 mmol) was added and the mixture was stirred for 24–48 h at room
temperature (TLC). Then, it was poured into crushed ice and extracted
with CH3COOC2H5. The extract was washed with H2O, brine, and
dried over Na2SO4. The solvent was evaporated under reduce pressure
to afford the impure product. The impure product was then purified by
flash chromatography (elution with CH3COOC2H5: petroleum ether =
4:1–8:1) to give 7a–j (45–55%).
This work was supported by the Fundamental Research Funds
for Central University, Southwest University for Nationalities
(No.11ZYXS17).
Methyl 3α-(2-(4-amino-3-(3-chlorophenyl)-5-thioxo-4,5-dihydro-
1,2,4-triazol-1-yl-)acetoxy)-12α-hydroxy-cholanate (7g): White solid,
yield 98%, m.p. 165–167 ºC; [α]2D0–107.1 (c 0.14, CH2Cl2); IR (cm1 ):
3341, 3260, 2937, 2867, 1731, 1572, 1450, 1384, 1299, 1189, 1094,
977, 889; 1H NMR (DMSO-d6, 400 MHz): δ 8.07 (s, 1H, ArH), 7.94
(dd, J = 1.6 Hz and J =1.6 Hz, 1H, ArH), 7.59–7.53 (m, 2H, ArH),
6.25 (s, 2H, NH2), 4.69–4.64 (m, 1H, 3β-H), 4.25 (s, 1H, 12β-H), 4.07
(s, 2H, –OCOCH2N), 3.78 (s, 1H, 12α-OH), 3.57 (s, 3H, –COOCH3),
0.90 (d, J = 6.4 Hz, 3H, 21-CH3), 0.86 (s, 3H, 19-CH3), 0.59 (s, 3H,
18-CH3); ESI-MS m/z (%): 1367 ([2M+H]+, 100). Anal. Calcd for
C35H49ClN4O5S: C, 62.43; H, 7.34; N, 8.32. Found: C, 62.72; H, 7.38;
N, 8.29%.
Methyl 3α-(2-(4-amino-3-(4-chlorophenyl)-5-thioxo-4,5-dihydro-
1,2,4-triazol-1-yl)-acetoxy)-12α-hydroxy-cholanate (7h): White solid,
yield 95%, m.p. 160–162 ºC; [α]2D0–64.9 (c 0.13, CH2Cl2); IR (cm−1):
3324, 3196, 2936, 2869, 1732, 1632, 1462, 1383, 1296, 1186, 1093,
998, 885, 836; 1H NMR (DMSO-d6, 400 MHz): δ 8.01 (d, J = 8.4 Hz,
2H, ArH), 7.60 (d, J = 8.4 Hz 2H, ArH), 6.23 (s, 2H, NH2), 4.66–4.64
(m, 1H, 3β-H), 4.26 (s, 1H, 12β-H), 4.07 (s, 2H, –OCOCH2N), 3.87
(s, 1H, 12α-OH), 3.57 (s, 3H, –COOCH3), 0.90 (d, J = 6.0 Hz, 3H,
21-CH3), 0.86 (s, 3H, 19-CH3), 0.59 (s, 3H, 18-CH3); ESI-MS m/z
(%): 1367 ([2M+H]+, 100). Anal. Calcd for C35H49ClN4O5S: C, 62.43;
H, 7.34; N, 8.32. Found: C, 62.61; H, 7.30; N, 8.35%.
Methyl 3α-(2-(4-amino-3-(4-bromophenyl)-5-thioxo-4,5-dihydro-
1,2,4-triazol-1-yl)-acetoxy)-12α-hydroxy-cholanate (7i): White solid,
yield 92%, m.p. 99–101 ºC; [α]2D0–129.8 (c 0.14, CH2Cl2); IR (cm−1):
3336, 3195, 2927, 2865, 1729, 1630, 1453, 1380, 1297, 1171, 980,
831, 724; 1H NMR (DMSO-d6, 400 MHz): δ 7.96 (d, J = 8.4 Hz, 2H,
ArH), 7.74 (d, J = 8.4 Hz, 2H, ArH), 6.23 (s, 2H, NH2), 4.70–4.65
(m, 1H, 3β-H), 4.26 (s, 1H, 12β-H), 4.07 (s, 2H, –OCOCH2N), 3.78
(s, 1H, 12α-OH), 3.58 (s, 3H, –COOCH3), 0.92 (d, J = 6.4 Hz, 3H,
21-CH3), 0.87 (s, 3H, 19-CH3), 0.60 (s, 3H, 18-CH3); ESI-MS m/z
(%): 1459 ([2M+Na]+, 100).Anal. Calcd for C35H49BrN4O5S: C, 58.57;
H, 6.88; N, 7.81. Found: C, 58.83; H, 6.85; N, 7.85%.
Received 22 March 2012; accepted 31 March 2012
Paper 1201226 doi: 10.3184/174751912X13364636691591
Published online: 26 June 2012
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