Synthesis and pharmacological activity of amides and the ozonolysis product of maleopimaric acid

Article abstract of DOI:10.1134/S1068162010060130

The synthesis of a new group of maleopimaric acid amides containing fragments of methyl ethers of amino acids, aliphatic amines, imidazole, and N-methylpiperazine was carried out. The ozonolysis of methylmaleopimarate occurs via the cleavage of the double

Full text of DOI:10.1134/S1068162010060130

ISSN 1068ꢀ1620, Russian Journal of Bioorganic Chemistry, 2010, Vol. 36, No. 6, pp. 762–770. © Pleiades Publishing, Ltd., 2010.
Original Russian Text © O.B. Kazakova, E.V. Tret’yakova, O.S. Kukovinets, G.A. Tolstikov, T.I. Nazyrov, I.V. Chudov, A.F. Ismagilova, 2010, published in Bioorganicheskaya
Khimiya, 2010, Vol. 36, No. 6, pp. 832–840.
Synthesis and Pharmacological Activity of Amides and the Ozonolysis
Product of Maleopimaric Acid
1
O. B. Kazakovaa, , E. V. Tret’yakovaa, O. S. Kukovinetsa, G. A. Tolstikova,
T. I. Nazyrova, I. V. Chudovb, and A. F. Ismagilovab
a Institute of Organic Chemistry, Ufa Scientific Center, Russian Academy of Sciences, pr. Oktyabrya 71, Ufa, 450054 Russia
b Bashkir State Agrarian University, Ufa, Russia
Received December 21, 2009; in final form, May 4, 2010
Abstract—The synthesis of a new group of maleopimaric acid amides containing fragments of methyl ethers
of amino acids, aliphatic amines, imidazole, and
Nꢀmethylpiperazine was carried out. The ozonolysis of
methylmaleopimarate occurs via the cleavage of the double bond C18(19) and the opening of an anhydrous
ring with the formation of secotriacid. As a result of the screening of the antiꢀinflammatory and antiulcer
activity of maleopimaric acid derivatives, new effective compounds such as maleopimaric acid and its methyl
ether, a product of ozonolysis—diterpenic secotriacid—and maleopimaric acid amide with Lꢀleucine were
found. An important advantage of the studied compounds is the low toxicity and the presence of bidirectional
activity in the absence of adverse effects on the animal.
Keywords: levopimaric acid, maleopimaric acid, amides, ozonolysis, antiulcer activity, antiꢀinflammatory activity
DOI: 10.1134/S1068162010060130
1
INTRODUCTION
zole, and Nꢀmethylpiperazine (scheme). The use of a
quantity of amine equimolar to the acid allowed us to
conduct the reaction selectively by the carboxyl funcꢀ
tion without involving the anhydrous ring.
Maleopimaric acid (II), which is the adduct of
diene synthesis of levopimaric acid (I) with maleic
anhydride, is an accessible product for chemical modꢀ
ification [1–8]. The processes of the oxidation and
regroupings of maleopimaric and the fumaropimaric
acids associated with them were studied in [9–14].
Methods of the selective decarboxylation, including
catalytic methods accompanied by a reaction of retroꢀ
diene synthesis, were developed [15]. The fungicidal
One of the effective methods of the oxidation of
double bonds is ozonolysis. The reaction of methyl
maleopimarate (III) with ozone was mentioned in the
work by L. Ruzicka [23]. We found that the ozonation
of the methyl ether of maleopimaric acid (III) in
methylene–methanol at 0°С produces a good yield of
secotriacid (XV). The structure of the obtained acid
was confirmed by the synthesis and determination of
the physicochemical characteristics of its full methyl
ether (XVI). The opening of the anhydrous ring of
methylmaleopimarate with the formation of two carꢀ
boxyl groups during ozonation probably occurs due to
the participation of the anhydrous ring in the stabilizaꢀ
tion of the peroxide products of the interaction of
ozone with the C18–C19 double bond spatially close
to it. It should be noted that the aldehyde group, which
forms in the C1 position during the cleavage of the
C18–C19 double bond by ozone, is very easily oxiꢀ
dized to the carboxyl group. The compound (XV) can
be obtained also during the ozonolysis of the trimethyl
ether of fumaropimaric acid but with a low yield and
after the chromatographic purification of the multiꢀ
component mixture [24].
properties of salts and ethers of Nꢀreplaced imides of
maleopimaric acid were revealed [16]. Compounds
with antiꢀinflammatory and heptaprotective activities
were found among the amides of maleopimaric acid
with piperazine, 4ꢀmethylꢀ and 4ꢀhydroxyethylpiperꢀ
azine, morpholine, and dimethylcarbamoyl.
While continuing the studies of the pharmacologiꢀ
cal properties of adducts of levopimaric acid [18–22],
we modified maleopimaric acid (II) at the carboxyl
group and bridging double bond and studied the antiꢀ
ulcer and antiꢀinflammatory activities of the obtained
compounds.
RESULTS AND DISCUSSION
A new group of amides (V)–(XIV) were synthesized
by the interaction of maleopimaryl chloride (IV) with
1
A study of the acute toxicity of the derivatives of
maleopimaric acid (II), (III), and (V)–(XV) showed
(see Experimental section) that their mean lethal dose
is within 8000 to 12000 mg/kg. The introduction of
ethers of amino acids, aliphatic amines, Hꢀimidaꢀ
1
Corresponding author; phone/fax: (347) 235ꢀ60ꢀ66; eꢀmail:
obf@anrb.ru.
762

SYNTHESIS AND PHARMACOLOGICAL ACTIVITY OF AMIDES
763
O
H
O
O
O
12
11
14
16
O
19 13
15
O
O
C
C
18
17
HC
HC
[2]
8
5
10
3
+
O
7
6
9
1
2
ii
H
4
O
COR
(I)
20
COR
COOH
–
3' 4'
2'
1'
(
(
(
II) R = OH,
III) R = OCH
IV) R = Cl
iii
i
(
(
(
(
(
(
V) R = NHCH(CH )CO CH
3 2 3
2' 1'
3
4' 3'
iv
VI) R = NH(CH ) CO CH
2 2
2' 1'
2
3
3'
4'
5' 6'
VII) R = NHCH(CO CH )CH(CH )
2
2' 1'
3
3 2
O
3'
4' 5'6'
VIII) R = NHCH(CO CH )CH(CH )
2
3
4' 5'
3 2
3' 2' 1'
6'7'
COOR
3
IX) R = NHCH(CO CH )CH CH(CH )
4
2
1
2
3
4'5'
2
3
4a
3' 2' 1'
6'
5
8
COOR
COOR
X
) R = NHCH(CO CH )(CH ) SCH
3
6
7
4b 10a
10
2
2 2
3
3' 5'–10'4'
2'
1'
9
(
XI) R = NHCH(Ph)CH CO CH
2 2 3
1' 2'–5' 6'
COOCH3
XV) R = H
XVI) R = CH
(
XII) R = NHCH(CH ) CH
2 4
3
(
(
2'
iii
1'
3
(
XIII) R =
XIV) R =
N
N
3'
N
1' 2'
(
NCH3
4'
3'
Reaction conditions: i. (COCl) , CHCl , 3 h; ii. NH CH(CH )CO CH · HCl for (V), NH (CH ) CO CH · HCl
2 2 2 2 3
2
3
2
3
2
3
for (VI), (L)ꢀNH CH(CO CH )CH(CH ) · HCl for (VII), (DL)ꢀNH CH(CO CH )CH(CH ) · HCl for (VIII),
2 2 3 3 2 2 2 3 3 2
NH CH(CO CH )CH CH(CH ) · HCl for (IX), (
L
)ꢀNH CH(CO CH )(CH ) SCH · HCl for (X),
2
2
3
2
3 2
2 2 3 2 2 3
NH CH(Ph)CH CO CH · HCl for (XI), NH CH(CH ) CH for (XII), NH C H N for (XIII),
2
2
2
3
2
2 4
3
2 3 3 2
NH C H N for (XIV), Et N, CHCl ; iii. CH N , Et O; iv. O , CH Cl ꢀMeOH, 0
°C.
2
5
11
2
3
3
2
2
2
3
2
2
Scheme.
doses close to LD16 to the stomachs of mice was gens of a nonprotein nature: silver nitrate and formaꢀ
accompanied by an insignificant inhibition in the lin. Ortophen was used for comparison. The data from
experimental animals and a decrease in their locomoꢀ Table 1 show that maleopimaric acid (II) manifests
tive activity (“freezing” on the spot). However, these antiꢀinflammatory activity more pronouncedly as
signs disappeared within 1 to 2 h and the physiological compared to ortophen, but less pronouncedly as comꢀ
activity was completely restored. After the administraꢀ pared to the product of ozonolysis (XV) in a dose of
tion of doses close to LD50, a shortꢀterm increase in 50 mg/kg.
the motor activity of mice from experimental groups
The derivatives of maleopimaric acid (V)–(XIV)
was registered for 20–30 min, after which inhibition
followed, which lasted from 6 to 12 h. The intragastric
introduction of absolutely lethal doses of compounds
have higher antiꢀinflammatory action than the acid
itself (Table 1). The highest activity in this group is
demonstrated by the amide of maleopimaric acid with
methyl ether of Lꢀleucine (IX) during argentonitrate
inflammation (within doses of 25 to 100 mg/kg).
(
II), (III), and (V)–(XV) caused the quick (within 5–
10 min) depression of motor activity and a lethal outꢀ
come within 12 to 24 h. The variability coefficient of
the lethal doses was 1.6–2.1, which indicates their sigꢀ
nificantly wide range. Thus, according to GOST
An interesting manifestation of the antiphlogistic
activity was registered in compounds (VII), (VIII),
12.1.00.7ꢀ76, compounds (II), (III), and (V)–(XV)
belong to class 4 of danger (low toxicity).
which contain fragments of
established that the presence of a fragment of DL
Lꢀ and DLꢀvaline. It was
ꢀ
The antiꢀinflammatory properties of derivatives of valine decreases the pharmacological effectiveness
maleopimaric acid were studied in three species of during the acute stages of the inflammatory reaction as
experimental inflammations induced by carrageenin, compared with compound (VII). At the same time, the
which is a phlogogen of a protein nature, and phlogoꢀ degree of the antiphlogistic activity of compound
RUSSIAN JOURNAL OF BIOORGANIC CHEMISTRY Vol. 36
No. 6
2010

764
KAZAKOVA et al.
Table 1. Antiꢀinflammatory activity* of compounds (II), (III), and (V)–(XV)
Carrageenin inflammation, the dose, mg/kg
50
Compound
25
25
25
100
100
100
A
B
A
B
A
B
(
(
(
(
(
(
(
(
(
(
(
(
(
II
III
)
38.7 1.9
37.6 1.9
40.3 1.7
38.2 1.6
37.9 1.6
52.8 3.3
38.2 1.6
42.1 1.6
44.7 1.7
39.7 2.5
42.3 2.5
41.1 1.7
39.4 2.5
40.7 1.3
54.5 1.1
29.1 1.4
31.0 1.5
26.0 1.1
30.0 1.3
30.4 1.3
3.1 0.2
30.0 1.3
22.9 0.9
18.1 0.7
27.2 1.7
22.3 1.3
24.6 1.0
27.7 1.7
25.4 0.8
n.s.
40.6 2.5
38.8 2.5
39.7 1.9
37.8 1.9
36.3 2.1
52.2 2.1
37.9 2.1
41.8 1.9
43.3 2.0
40.1 1.8
44.3 1.8
40.8 1.8
34.5 1.7
40.7 1.3
54.5 1.1
25.5 1.6
28.9 1.9
27.2 1.3
30.6 1.5
33.4 1.9
4.2 0.2
30.4 1.7
23.3 1.0
20.6 0.9
26.4 1.2
18.7 0.8
25.2 1.1
36.7 1.8
25.4 0.8
n.s.
40.0 2.0
35.9 1.8
41.1 2.0
37.2 1.8
39.1 1.8
53.2 2.6
37.3 1.8
42.3 1.9
44.6 2.3
39.1 2.0
45.3 1.9
49.4 2.0
34.7 2.2
40.7 1.3
54.5 1.1
26.6 1.3
34.2 1.7
24.5 1.1
31.8 1.5
28.3 1.3
2.5 0.1
31.6 1.5
22.5 1.0
18.3 1.0
28.2 1.5
16.8 0.7
9.4 0.4
36.4 2.3
25.4 0.8
n.s.
)
V)
VI
VII
VIII
)
)
)
IX
)
X)
XI
)
XII
XIII
XIV
XV
)
)
)
)
Ortophen
Control
Argentonitrate inflammation, the dose, mg/kg
50
Compound
A
B
A
B
A
B
(
(
(
(
(
(
(
(
(
(
(
(
(
II
III
V
VI
VII
VIII
IX
X
)
35.2 2.2
33.7 2.2
35.4 2.0
34.2 1.9
29.3 1.4
34.8 1.4
26.2 1.3
31.9 1.9
37.2 1.8
36.8 1.9
37.5 1.8
35.7 1.8
29.7 1.8
36.2 0.1
43.1 0.2
18.2 1.1
21.8 1.4
17.7 1.0
20.7 1.1
31.9 1.6
19.3 0.8
39.3 1.9
26.0 1.6
13.7 0.6
14.5 0.7
13.0 0.6
17.2 0.9
31.1 1.9
15.9 0.1
n.s.
34.3 1.9
31.6 1.5
35.8 1.4
34.6 1.5
28.1 1.8
32.6 1.7
25.9 1.4
33.7 1.4
37.9 1.7
37.2 1.9
38.0 1.7
36.2 1.7
27.8 1.9
36.2 0.1
43.1 0.2
20.4 1.2
26.7 1.2
16.9 0.7
19.6 0.9
34.7 2.2
24.2 1.3
39.8 2.2
21.8 0.9
12.1 0.6
13.7 0.7
11.6 0.5
16.0 0.8
35.6 2.5
15.9 0.1
n.s.
34.0 1.9
34.0 0.7
37.2 1.3
35.0 1.8
30.2 1.5
31.4 1.2
26.3 1.4
33.8 1.8
37.5 1.9
38.0 1.9
37.8 1.9
36.2 1.6
27.6 1.2
36.2 0.1
43.1 0.2
21.0 1.1
21.1 0.4
13.7 0.5
18.8 0.9
29.8 1.5
27.2 1.1
38.9 2.1
21.5 1.1
12.8 0.6
11.6 0.6
12.3 0.6
16.0 0.7
35.8 1.6
15.9 0.1
n.s.
)
)
)
)
)
)
)
XI
)
XII
XIII
XIV
XV
)
)
)
)
Ortophen
Control
Formalin inflammation, the dose, mg/kg
50
Compound
A
B
A
B
A
B
(
(
(
(
(
(
(
(
(
(
(
(
(
II
III
)
26.1 0.8
24.1 1.1
26.1 0.9
21.2 1.0
26.1 1.0
22.0 0.7
23.3 1.2
27.8 1.3
25.4 1.1
26.6 1.2
28.1 1.0
25.9 1.1
22.9 0.8
26.1 0.9
35.8 0.2
27.2 0.9
32.5 1.5
27.0 1.0
40.7 1.9
27.0 1.1
38.5 1.2
35.0 1.8
22.3 1.1
29.1 1.3
25.6 1.1
21.6 0.8
27.6 1.2
37.7 1.4
27.0 0.9
n.s.
25.3 1.2
23.9 1.0
25.3 1.3
22.6 1.3
22.7 1.2
22.3 1.3
22.2 1.2
24.9 1.3
25.4 1.0
27.0 1.1
28.2 1.5
26.4 1.5
20.7 0.7
26.1 0.9
35.8 0.2
29.5 1.4
33.2 1.4
29.4 1.6
36.8 2.2
36.5 1.9
37.5 2.3
38.0 2.0
30.6 1.6
28.9 1.2
24.5 1.0
21.3 1.1
26.2 1.5
42.2 1.6
27.0 1.0
n.s.
23.7 1.2
23.6 1.5
25.4 0.8
23.1 1.0
22.8 1.2
23.6 1.0
22.3 0.9
23.4 0.9
25.8 0.9
27.0 1.1
28.1 1.1
26.1 1.0
23.6 1.5
26.1 0.9
35.8 0.2
33.7 1.7
33.9 2.2
29.0 0.9
35.3 1.6
36.4 1.9
34.1 1.5
37.6 1.5
34.6 1.3
27.9 1.0
24.6 1.0
21.6 0.9
27.1 1.0
34.1 2.1
26.9 1.0
n.s.
)
V)
VI
VII
VIII
)
)
)
IX
)
X)
XI
)
XII
XIII
XIV
XV
)
)
)
)
Ortophen
Control
* Was estimated (in per cents) by the increase (A) and depression (B) in the inflammatory edema; (n.s.) not studied.
RUSSIAN JOURNAL OF BIOORGANIC CHEMISTRY Vol. 36
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SYNTHESIS AND PHARMACOLOGICAL ACTIVITY OF AMIDES
765
(
VIII) can be brought to a level comparable with that The optical absorption spectra were measured on a
of derivative (VII) by increasing the dose from 50 to PerkinꢀElmer 241 MC polarimeter (Germany) in a
100 mg/kg.
The amide of maleopimaric acid with
demonstrates higher antiꢀinflammatory activity on the
model of carrageenin inflammation compared with
tube 1ꢀdm long. An Ozonꢀ2K ozonizer (Russia) was
used for ozonation. TLC analysis was conducted on
Silufol plates (Chemapol, Czech Republic) using a
20 : 1 chloroform–methanol system of solutions; the
compound was detected with a 5% solution of phosꢀ
βꢀalanin (VI)
orthophen. The activity of amide with Lꢀalanin (V) is
photungstic acid in ethanol (2–3 min at 100–120
Compounds (II and (III) were obtained according to
[2].
°
С).
comparable with that of the comparison preparation
within the range of the effective doses from 25 to
100 mg/kg on all models of inflammation. Compound
)
(
VI) showed higher activity as compared with that of
compound ( ), but less pronounced as compared with
compounds (VII and (VIII).
(5R,9R,13R,17R)ꢀ5,9ꢀDimethylꢀ19ꢀ(1ꢀmethylethyl)ꢀ
15ꢀoxaꢀ14,16ꢀdioxopentacyclo[10.5.2.01,10.04,9.013,17]hepꢀ
tadecꢀ18ꢀeneꢀ5ꢀcarbonyl chloride (maleopimaryl
chloride) (IV). Oxalyl chloride (0.3 ml, 3.5 mmol) was
V
)
Thus, compounds (II), (III), and (XV) proved to be
the most active with steadily high antiphlogistic
action, while maleopimaric acid amides with methyl
added dropwise while stirring to compound (II
)
(1 mmol, 0.42 g) in dry chloroform (10 ml). The mixꢀ
ture was kept under nitrogen for 4 h. The solvent was
evaporated in the vacuum of a waterꢀjet pump. Yield
0.40 g (95%). Found, %: C 72.09; H 7.92; Cl 8.06.
ethers of
L
ꢀmethionine (
X
) and
L
ꢀphenylꢀ
β
ꢀalanine
(
XI), and heterocyclic residues (XIII
)
and (XIV)
proved to be the least active relative to the comparison
preparation.
C24H31ClO4 (M 418.95). Calculated, %: C 72.45; H
The results of the studies on the antiulcer properꢀ
8.18; Cl 8.23.
ties of maleopimaric acid (II) and its derivatives (III
and ( )–(XV) on two models of experimental ulcers
)
Synthesis of compounds (V)–(XIV). A solution of
one of the following compounds—hydrochloride of
V
are given in Table 2 and indicate the ability to depress
the process of the formation of ulcerations of mucous
membranes of the gastrointestinal tract under the
influence of unfavorable factors. The highest level of
antiulcer activity was noted in compound (XV) in a
dose of 25 mg/kg. An increase in the dose of comꢀ
pound (XV) from 25 to 100 mg/kg on the model of
indometacin ulcers did not lead to an improvement of
this property, whereas the positive tendency of increasꢀ
ing the activity was noted during the induction of
ulcers with acetic acid by increasing the dose to
the methyl ether of amino acid (
ꢀvaline, DLꢀvaline, ꢀleucine,
ꢀphenylꢀ ꢀalanine, respectively), hexylamine, 1
imidazole, or ꢀmethylpiperazine (1.1 mmol)—in
L
ꢀalanine,
β
ꢀalanine,
L
L
L
Lꢀmethionine, and
β
Hꢀ
N
dry chloroform (5 ml) was added while stirring to a
solution of (IV) (1 mmol, 0.43 g) in dry chloroform
(15 ml). The solvent was evaporated in the vacuum of
a waterꢀjet pump. The reaction product was purified
by Al2O3 column chromatography (chloroform as the
eluent).
Nꢀ[(5
R,9R,13R,17R)ꢀ5,9ꢀDimethylꢀ19ꢀ(1ꢀmethꢀ
100 mg/kg. Compounds (II), (III), (V), and (VII)–(IX)
ylethyl)ꢀ15ꢀoxaꢀ14,16ꢀdioxopentacyclo[10.5.2.01,10
.
showed activity on the model of indometacin ulcers at
a level of ~2.59 at a minimum dose from 50 to
100 mg/kg. At the same time, for acetic ulcers, the
activity at level
ric acid (II) and its methyl ether (III).
Maleopimaric acid amides (VI
showed insufficiently pronounced antiulcer activity.
04,9.013,17]heptadecꢀ18ꢀeneꢀ5ꢀcarbonyl]ꢀ
methylether (V). Yield 0.34 g (79%).
L
ꢀalanine
Rf 0.63. Mp 176–
≥2 was registered only for maleopimaꢀ
20
178
°
С. [α]D – 23
°
(с
0.05, CHCl3). Found, %: C
485.61). Calcuꢀ
lated, %: C 69.25; H 8.09; N 2.88. 1H NMR spectrum:
0.59 (3 Н, s, СН3), 0.99 and 1.00 (6 H, both d, 6.8,
68.97; H 7.86; N 2.65. C28H39NO6 (
M
)
and (X)–(XIV)
J
Thus, maleopimaric acid (II) and its derivatives
III), (IX), and (XV) show some promise for further
2СН3), 1.15 (3 H, s, СН3), 1.15–1.30 (1 H, m, H4),
1.35–1.80 (15 H, m, CH2, CH), 2.23 (1 H, m, CH),
(
study as antiꢀinflammatory and antiulcer agents. An
important advantage of the studied compounds comꢀ
pared with the existing drugs of unidirectional action is
the twoꢀway (combined) (antiꢀinflammatory and antiꢀ
ulcer) activity and the absence of negative influences
on the animal’s body characteristic of groups of drugs
with a similar manifestation of activity.
2.51 (1 H, dt,
3.08 (1 H, dd,
(3 H, s, OMe), 4.35 (1 H, m, H3'), 5.53 (1 H, br. s,
H18), 6.49 (1 H, dd,
5.4, J 1.2, NH). 13C NMR specꢀ
J
3,
J
14, H10), 2.71 (1 H, d,
J11.0, H12),
J
3,
J11, H17), 3.09 (1 H, m, H13), 3.69
J
trum: 15.5, 16.6 (C4'), 17.0, 19.9, 20.5, 20.9, 27.1,
32.7, 33.5, 34.6, 35.2, 35.6, 36.7, 37.7, 37.8, 40.3,
45.6, 46.5, 49.6, 51.7 (C1'), 53.0, 53.1 (C3'), 125.1
(C18), 147.9 (C19), 170.9 (C14), 172.7 (C16), 173.3
(C2'), 178.3 (C20).
EXPERIMENTAL
The 1Нꢀ and 13C NMR spectra were recorded on a
Nꢀ[(5R,
ethyl)ꢀ15ꢀoxaꢀ14,16ꢀdioxopentacyclo[10.5.2.01,10.04,9.
013,17]heptadecꢀ18ꢀeneꢀ5ꢀcarbonyl]ꢀ
ꢀalanine methyl
ether (VI). Yield 0.34 g (78%). f 0.68. Mp 168–170 С.
9R,13R,17R)ꢀ5,9ꢀDimethylꢀ19ꢀ(1ꢀmethylꢀ
Bruker AMꢀ300 spectrometer (Germany, 300 and
75.5 MHz, respectively, , ppm, SSCC, Hz) in CDCl3
using tetramethylsilane as the internal standard. The
melting points were determined on a Boetius plate.
δ
β
R
°
RUSSIAN JOURNAL OF BIOORGANIC CHEMISTRY Vol. 36
No. 6
2010

766
KAZAKOVA et al.
Table 2. Antiulcer activity of compounds (II), (III), and (V)–(XV)
Type of experimental ulcers
Indometacin
Acetic acid
Compound Dose, mg/kg
mean number of deꢀ
mean number of deꢀ
antiulcer activity
antiulcer activity
structions
structions
(
(
(
(
(
(
(
(
(
(
(
(
(
II
)
25
50
3.9 0.4
3.7 0.3
4.2 0.2
5.6 0.4
4.7 0.4
4.6 0.4
6.6 0.5
7.6 0.4
7.3 0.5
8.6 0.5
7.7 0.5
7.7 0.5
6.2 0.5
4.9 0.4
6.9 0.5
5.9 0.4
6.1 0.4
6.1 0.5
5.2 0.3
5.2 0.4
5.1 0.3
7.2 0.3
7.4 0.5
7.6 0.3
5.6 0.4
6.1 0.4
6.3 0.5
7.6 0.3
7.8 0.3
8.0 0.3
8.3 0.4
8.3 0.2
8.6 0.4
9.2 0.3
8.5 0.2
8.8 0.4
3.2 0.2
3.7 0.2
3.5 0.2
14.4 0.8
3.7 0.1
3.9 0.1
3.4 0.1
2.6 0.1
3.1 0.1
3.2 0.1
2.2 0.1
1.9 0.1
2.0 0.1
1.7 0.1
1.9 0.1
1.9 0.1
2.5 0.9
3.1 0.9
2.1 0.1
2.4 0.1
2.4 0.1
2.4 0.1
2.8 0.1
2.8 0.1
2.8 0.1
2.0 0.1
1.9 0.1
1.9 0.1
2.6 0.1
2.4 0.1
2.3 0.1
1.9 0.1
1.9 0.1
1.8 0.1
1.7 0.1
1.7 0.1
1.7 0.1
1.6 0.1
1.7 0.1
1.6 0.1
4.4 0.1
3.9 0.2
4.1 0.1
13.3 0.9
12.9 1.1
13.6 0.8
13.3 0.8
14.1 1.1
14.4 1.2
15.7 1.1
14.7 1.2
16.1 1.5
18.0 1.1
18.4 1.1
17.9 1.2
16.7 1.1
15.0 1.1
16.3 1.1
14.6 1.2
13.7 1.0
14.3 0.9
14.6 1.0
13.8 0.9
14.7 0.9
17.5 1.3
17.2 0.9
18.6 1.2
20.3 0.8
19.8 0.8
20.9 1.0
19.7 0.9
19.9 0.9
19.7 0.6
20.3 0.7
19.8 0.8
20.1 0.8
17.3 1.6
14.7 0.9
14.0 0.9
12.6 1.0
12.3 0.9
11.2 0.7
28.3 1.3
2.1 0.1
2.2 0.1
2.1 0.1
2.1 0.1
2.0 0.1
1.9 0.1
1.8 0.1
1.9 0.1
1.7 0.1
1.6 0.1
1.5 0.1
1.6 0.1
1.7 0.3
1.9 0.3
1.7 0.1
1.9 0.1
2.1 0.1
2.0 0.1
1.9 0.1
2.1 0.1
1.9 0.1
1.6 0.1
1.6 0.1
1.5 0.1
1.4 0.1
1.4 0.1
1.4 0.1
1.4 0.1
1.4 0.1
1.4 0.1
1.4 0.1
1.4 0.1
1.4 0.1
1.6 0.1
1.9 0.1
2.0 0.1
2.2 0.1
2.3 0.1
2.5 0.1
100
25
III
)
50
100
25
VI
)
50
100
25
X
)
)
50
100
25
V
50
100
25
IX
)
50
100
25
VII
)
50
100
25
XI
)
50
100
25
VIII
)
50
100
25
XIV
)
50
100
25
XIII
)
50
100
25
XII
)
50
100
25
XV
)
50
100
Control
n.s.
*
n.s.*
n.s.*
* (n.s.) not studied.
RUSSIAN JOURNAL OF BIOORGANIC CHEMISTRY Vol. 36
No. 6
2010

SYNTHESIS AND PHARMACOLOGICAL ACTIVITY OF AMIDES
767
20
[α]D – 16
°
(
с
0.05, CHCl3). Found, %: C 68.93; H 04,9.013,17]heptadecꢀ18ꢀeneꢀ5ꢀcarbonyl]ꢀ
L
ꢀleucine
methyl ether (IX). Yield 0.35 g (81%).
Rf 0.72. Mp
8.01; N 2.70. C28H39NO6 (
69.25; H 8.09; N 2.88. 1H NMR spectrum: 0.60 (3 H,
с, CH3), 0.99 and 1.01 (6 H, both d, 6.8, 2СH3), 1.15
(3 H, s, CH3), 1 (1 H, m, H4), 1.34–1.80 (15 H, m,
СН2, CH), 2.23 (1 H, m, CH), 2.51 (1 H, dt,
H10), 2.70 (1 H, d, 11, H12), 3.05 (1 H, dd,
H17), 3.10 (1 H, m, H13), 3.70 (3 H, s, OMe), 4.52 (1
H, m, H3'), 5.54 (1 H, br.s, H18), 6.50 (1 H, dd, 5.4
1.2, NH). 13C NMR spectrum: 15.5, 16.4, 17.0, 19.9,
20.4, 20.7, 26.9, 32.7, 33.5 (C3'), 34.7, 35.2, 35.6
(C4'), 36.7, 37.5, 37.8, 40.8, 45.2, 45.8, 49.6, 51.9
(C1'), 53.0, 53.2, 125.1 (C18), 148.1 (C19), 170.8
(C14), 172.7 (C16), 172.9 (C2'), 178.2 (C20).
M 485.61). Calculated, %: C
20
117–119
°
С. [α]D + 24
°
(
с
0.05, CHCl3). Found, %: C
527.69). Calcuꢀ
lated, %: C 70.56; H 8.59; N 2.65. 1H NMR spectrum:
0.59 (3 H, s, CH3), 0.97 and 0.99 (6 H, both d, 6.8,
J
70.85; H 8.24; N 2.76. C31H45NO6 (
M
J
3,
J
14,
J
J
J
3,
J
11,
2СH3), 1.15 (3 H, s, CH3), 1.18–1.30 (1 H, m, H4),
1.37–1.82 (21 H, m, CH2, CH), 2.23 (1 H, m, CH),
J
,
2.51 (1 H, dt,
3.08 (1 H, dd,
J
3,
3,
J
14, H10), 2.71 (1 H, d,
J 11, H12),
J
J
J 11, H17), 3.10 (1 H, m, H13), 3.70
(3 H, s, OMe), 4.51 (1 H, m, H3'), 5.49 (1 H, br.s,
H18), 6.19 (1 H, dd,
J
5.4, J 1.2, NH). 13C NMR specꢀ
trum: 15.5, 16.6, 16.9, 19.8, 20.4, 20.9, 21.8 (C7'),
22.4 (C6'), 22.6, 24.9 (C5'), 27.1, 32.5, 34.4, 35.5,
36.7, 37.5, 37.6, 40.2, 41.3 (C4'), 45.5, 46.5, 49.2, 50.8
(C1'), 52.1 (C3'), 52.9, 125.1 (C18), 147.8 (C19),
Nꢀ[(5R,
9R,13R,17R)ꢀ5,9ꢀDimethylꢀ19ꢀ(1ꢀmethylꢀ
ethyl)ꢀ15ꢀoxaꢀ14,16ꢀdioxopentacyclo[10.5.2.01,10.04,9.
013,17] heptadecꢀ18ꢀeneꢀ5ꢀcarbonyl]ꢀ
L
ꢀvaline methyl 170.9 (C14), 172.7 (C16), 173.6 (C2'), 178.2 (C20).
ether (VII). Yield 0.40 g (92%).
Rf 0.75. Mp 129–
Nꢀ[(5R,9R,13R,17R)ꢀ5,9ꢀDimethylꢀ19ꢀ(1ꢀmethylꢀ
20
ethyl)ꢀ15ꢀoxaꢀ14,16ꢀdioxopentacyclo[10.5.2.01,10
.
131
°
С. [α]D + 44
°
(с
0.05, CHCl3). Found, %: C
513.63). Calcuꢀ
lated, %: C 70.15; H 8.44; N 2.73. 1H NMR spectrum:
0.60 (3 H, с, CH3), 0.97 and 1.00 (6 H, both d, 6.8,
04,9.013,17]heptadecꢀ18ꢀeneꢀ5ꢀcarbonyl]ꢀ
L
ꢀmethionꢀ
70.26; H 8.47; N 2.77. C30H43NO6 (
М
ine methyl ether (X). Yield 0.32 g (74%).
Rf 0.68. Mp
20
J
79–81
°
С. [α]D – 23
°
(с
0.05, CHCl3). Found, %: C
531.7).
2СH3), 1.14 (3 H, s, CH3), 1.18–1.33 (1 H, m, H4),
1.45–1.85 (19 H, m, CH2, CH), 2.25 (1 H, m, CH),
2.50 (1 H, dt,
3.08 (1 H, dd,
(3 H, s, OMe), 4.49 (1 H, m, H3'), 5.52 (1 H, br.s,
H18), 6.23 (1 H, dd, 5.4,
trum: 15.2, 16.0, 16.9, 19.7, 20.4, 21.0, 21.5 (C6'),
22.0 (C5'), 22.3, 25.2 (C4'), 27.8, 32.8, 34.6, 35.0,
36.9, 37.1, 37.6, 41.2, 45.1, 47.0, 49.9, 51.8 (C1'), 52.9
(C3'), 53.1, 125.9 (C18), 147.5 (C19), 170.5 (C14),
172.9 (C16), 173.3 (C2'), 178.1 (C20).
65.01; H 7.08; N 2.38; S 5.86. C29H41NSO6 (
M
1
Calculated, %: C 65.51; H 7.77; N 2.63. H NMR
spectrum: 0.61 (3 H, s, CH3), 0.99 and 1.01 (6 H, both
J
J
3,
3,
J
J
14, H10), 2.70 (1 H, d,
11, H17), 3.11 (1 H, m, H13), 3.71
J 11, H12),
d,
J 6.8, 2СH3), 1.16 (3 H, s, CH3), 1 (1 H, m, H4),
1.40–1.80 (17 H, m, СН2, CH), 2.23 (1 H, m, CH),
J
J
1.2, NH). 13C NMR specꢀ
2.51 (1 H, dt,
J
3,
J
14, H10), 2.72 (1 H, d,
11, H17), 3.09 (1 H, m, H13), 3.71
5.2, J 1.9, J 7.1, H3'),
5.53 (1 H, s, H18), 6.56 (1 H, dd,
5.4, J 1.2, NH). 13C
J 11, H12),
3.08 (1 H, dd,
(3 H, s, OMe), 4.69 (1 H, dt,
J
3,
J
J
J
NMR spectrum: 15.4, 15.5, 16.9, 18.9, 19.3, 19.8,
21.0, 27.0, 30.0 (C5'), 30.8 (C4'), 32.6, 34.5, 35.5,
36.8, 37.6, 37.7, 40.2, 45.5, 46.5, 49.4, 52.3 (C1'), 52.9
(C3'), 53.0, 125.0 (C18), 147.8 (C19), 170.7 (C14),
172.5 (C16), 176.0 (C2'), 178.1 (C20).
Nꢀ[(5R,9R,13R,17R)ꢀ5,9ꢀDimethylꢀ19ꢀ(1ꢀmethylꢀ
ethyl)ꢀ15ꢀoxaꢀ14,16ꢀdioxopentacyclo[10.5.2.01,10
.
04,9.013,17]heptadecꢀ18ꢀeneꢀ5ꢀcarbonyl]ꢀDLꢀvaline
methyl ether (VIII). Yield 0.32 g (74%). Rf 0.75. Mp
Nꢀ[(5R,
9R,13R,17R)ꢀ5,9ꢀDimethylꢀ19ꢀ(1ꢀmethylꢀ
20
ethyl)ꢀ15ꢀoxaꢀ14,16ꢀdioxopentacyclo[10.5.2.01,10
.
106–108
°С. [α]D + 3
°
(
с
0.05, CHCl3). Found, %: C
513.67). Calcuꢀ
lated, %: C 70.15; H 8.44; N 2.73. 1H NMR spectrum:
0.61 (3 H, s, CH3), 0.99 and 1.00 (6 H, both d, 6.8,
04,9.013,17]heptadecꢀ18ꢀeneꢀ5ꢀcarbonyl]ꢀ
Lꢀphenylalaꢀ
nine methyl ether (XI). Yield 0.33 g (76%). Rf 0.65. Mp
70.25; H 8.44; N 2.76. C30H43NO6 (
М
20
J
106–108
°
С. [α]D +10
°
(с
0.05, CHCl3). Found, %: C
561.71). Calcuꢀ
lated, %: C 72.70; H 7.72; N 2.49. 1H NMR spectrum:
0.59 (3 H, s, CH3), 0.99 and 1.00 (6 H, both d, 6.8,
2СH3), 1.15 (3 H, s, CH3), 1.15–1.29 (1 H, m, H4),
1.45–1.90 (19 H, m, CH2, CH), 2.25 (1 H, m, CH),
2.50 (1 H, dt,
3.08 (1 H, dd,
(3 H, s, OMe), 4.47 (1 H, m, H3'), 5.52 (1 H, br.s,
H18), 6.23 (1 H, dd, 5.4,
trum: 15.2, 16.0, 16.9, 19.8, 20.4, 21.0, 21.5 (C6'),
22.0 (C5'), 22.3, 25.2 (C4'), 27.8, 32.8, 34.4, 35.0,
36.9, 37.1, 37.6, 41.2, 45.0, 47.0, 49.9, 51.8 (C1'), 52.9
(C3'), 53.2, 125.9 (C18), 147.1 (C19), 170.0 (C14),
172.9 (C16), 173.1 (C2'), 178.0 (C20).
72.88; H 7.59; N 2.33. C34H43NO6 (
M
J
J
3,
3,
J
J
14, H10), 2.70 (1 H, d,
11, H17), 3.11 (1 H, m, H13), 3.70
J 11, H12),
J
2СH3), 1.15 (3 H, s, CH3), 1.17–1.31 (1 H, m, H4),
1.35–1.80 (12 H, m, CH2, CH), 2.23 (1 H, m, CH),
2.51 (1 H, dt,
3.05 (2 H, dd,
J
J
1.2, NH). 13C NMR specꢀ
J
J
3,
5.6,
J
14, H10), 2.71 (1 H, d,
J
11, H12),
3, 11,
J
6.5, H4'), 3.08 (1 H, dd,
J
J
H17), 3.12 (1 H, br.s, H13), 3.68 (3H, s, OMe), 4.75
(1 H, dt, 5.8, J 5.4, J 11.5, H3'), 5.50 (1 H, br.s, H18),
6.15 (1 H, dd, 5.4, J 1.2, NH), 6.95–7.12 (2 H, m,
J
J
13
H6', H10'), 7.13–7.30 (3 H, m, H7', H8', H9'). C
NMR spectrum: 15.4, 15.5, 16.4, 16.9, 19.8, 20.5,
Nꢀ[(5R,9R,13R,17R)ꢀ5,9ꢀDimethylꢀ19ꢀ(1ꢀmethylꢀ
ethyl)ꢀ15ꢀoxaꢀ14,16ꢀdioxopentacyclo[10.5.2.01,10.ꢀ 27.1, 20.8, 32.6, 34.5, 35.6, 36.8, 37.4 (C4'), 37.5,
RUSSIAN JOURNAL OF BIOORGANIC CHEMISTRY Vol. 36
No. 6
2010

768
KAZAKOVA et al.
37.8, 40.2, 45.6, 46.6, 48.9, 52.6 (C1'), 52.8, 53.1 (16 H, m, CH2, CH), 2.22 (1 H, dt,
J
3,
2.23 (1 H, m, CH), 2.43 (3 H, s, NꢀCH3), 2.68 (1 H, d,
11.0, H12), 3.09 (2 H, m), 3.68–3.89 (4 H, m, H1',
J 14, H10),
(C3'), 125.2 (C18), 127.1 (C8'), 128.5 (C10'), 128.6
(C6'), 129.1 (C9'), 129.0 (C7'), 135.8 (C5'), 147.8
(C19), 170.9 (C14), 172.2 (C16), 172.7 (C2'), 178.3
(C20).
J
H3'), 5.51 (1 H, br.s, H18). 13C NMR spectrum: 15.6,
16.5, 16.7, 18.5 (C5'), 20.0, 20.4, 22.5, 27.0, 32.6,
34.8, 35.5, 36.9, 37.7, 37.8, 40.3, 44.3, 45.6 (C3'), 45.7
(C4'), 47.0, 49.6, 53.0, 53.4, 53.5 (C2'), 54.2 (C1'),
125.3 (C18), 147.9 (C19), 170.9 (C14), 172.7 (C16),
177.4 (C20).
Nꢀ[(5R,
ethyl)ꢀ15ꢀoxaꢀ14,16ꢀdioxopentacyclo[10.5.2.01,10
04,9.013,17]heptadecꢀ18ꢀeneꢀ5ꢀcarbonyl]hexylamine
9R,13R,17R)ꢀ5,9ꢀDimethylꢀ19ꢀ(1ꢀmethylꢀ
.
(XII). Yield 0.27 g (64%). Rf 0.72. Mp 100–102
°С.
20
(4bS,
8
R
)
ꢀ
4
b
,8ꢀDimethylꢀ3ꢀisobutyrylperhydroꢀ
ꢀtetracarboxylic acid 8ꢀ
[α]D – 14
°
(с
0.05, CHCl3). Found, %: C 74.18; H
phenanthreneꢀ1,2,8,10
methyl ether (XV). Ozone was passed through a soluꢀ
tion of compound (III) (2 mmol, 0.84 g) in a 1 : 1 mixꢀ
a
9.16; N 2.74. C30H45NO4 (M 483.68). Calculated, %: C
74.50; H 9.38; N 2.90. 1H NMR spectrum: 0.59 (3 H,
s, CH3), 0.89 (3 H, s, H6'), 0.99 and 1.02 (6 H, both d,
ture of CH2Cl2 and MeOH at
0°С until the starting
J
6.8, 2СH3), 1.25 (3 H, s, CH3), 1.26–1.51 (9 H, m,
compound vanished (TLC). The mixture was kept at
room temperature for 3 h. The precipitate was recrysꢀ
tallized from chloroform. Yield 0.75 g (89%). Mp
H2', H3', H4', H5', H4), 1.52–1.80 (15 H, m, CH2
,
CH), 2.23 (1 H, m, CH), 2.51 (1 H, dt,
2.71 (1 H, d, 11, H12), 3.08 (1 H, dd,
J
J
3,
3,
J
J
14, H10),
11, H17),
J
220–222 С. Found, %: C 62.25; H 7.27. C25H36O9 (M
°
1
3.11 (1 H, m, H13), 3.46 (2 H, m, H1'), 5.53 (1 H, br.s,
H18), 5.70 (1 H, br.s, NH). 13C NMR spectrum: 13.9,
15.5 (C6'), 16.8, 17.0, 19.9, 20.4, 20.9 (C5'), 22.4, 26.5
(C3'), 27.1, 29.5 (C2'), 31.3 (C4'), 32.6, 35.4, 35.6,
36.8, 37.6, 37.8, 39.7 (C1'), 40.2, 45.6, 46.5, 49.7,
54.2, 55.0, 125.2 (C18), 147.8 (C19), 170.9 (C14),
172.7 (C16), 178.0 (C20).
480.55). Calculated, %: C 62.49; H 7.55. H NMR
spectrum: 0.71 (3 H, s, CН3), 1.05 and 1.08 (6 H, both
d,
.48–1.98 (10 H, m, CH2, CH), 2.00 (1 H, qu,
Н8 ), 2.27 (1 H, d, 11.8, H3), 2.39 (1 H, d, 11, H2),
2.78 (1 H, m), 2.92 (1 H, dt, 4.5, 11.8, H1), 3.75 (3
J
6.8, 2СH3), 1.09 (3 H, s, CН3), 1 (3 H, m, H4, H4
а),
1
J
11.8,
а
J
J
J
J
13
H, s, OMe). C NMR spectrum: 13.3, 15.2, 16.5,
18.5, 19.6, 21.8, 33.5, 35.6, 36.4, 37.0, 37.5, 42.4,
42.9, 46.3, 47.9, 49.1, 50.7, 55.2, 55.4, 165.8
(COOH), 169.6 (COOH), 173.1 (COOH), 177.5
(COOH), 210.7 (C=O).
1
ꢀ
[(5R,
ethyl)ꢀ15ꢀoxaꢀ14,16ꢀdioxopentacyclo[10.5.2.01,10
04,9.013,17]heptadecꢀ18ꢀeneꢀ5ꢀcarbonyl]ꢀ1
ꢀimidaꢀ
9R,13R,17R)ꢀ5,9ꢀDimethylꢀ19ꢀ(1ꢀmethylꢀ
.
H
zole (XIII). Yield 0.30 g (69%). Rf 0.66. Mp 149–
(4bS,
phenanthreneꢀ1,2,8,10
1,2,8,10 ꢀtetramethyl ether (XVI). An ether solution
of CH2N2 (70 ml) (TLC) was added to compound (XV
(1 mmol, 0.48 g). After recrystallization from methaꢀ
nol, the yield was 0.45 g (88%). Mp 104–107
Found, %: C 64.49; H 8.27. C28H42O9 ( 522.63). Calꢀ
culated, %: C 64.35; H 8.10. 1H NMR spectrum: 0.73
(3 H s, СН3), 1.08 and 1.11 (6 H, both d, 6.8, 2СH3),
1.13 (3 H, s, СН3), 1.10–1.40 (3 H, m, H4, H4 ), 1.50–
1.95 (10 H, m, СН2, CH), 2.05 (1 H, qu,
Н8 ), 2.30 (1 H, d, 12, H3), 2.41 (1 H, d,
2.80 (1 H, m), 2.94 (1 H, dt, 4.5,
8
R
)ꢀ4
b
,8ꢀDimethylꢀ3ꢀisobutyrylperhydroꢀ
20
151
°
С. [α]D – 5
°
(с
0.05, CHCl3). Found, %: C 71.72;
465.58). Calculated, %:
C 71.97; H 7.61; N 6.22. H NMR spectrum: 0.59 (3
H, s, CH3), 0.99 and 1.00 (6 H, both d, 6.8, 2СH3),
1.15 (3 H, s, CH3), 1 (1 H, m, H4), 1.80–1.35 (12 H,
m, CH2, CH), 2.23 (1 H, m, CH), 2.51 (1 H, dt, 3,
14, H10), 2.71 (1 H, d, 11, H12), 3.08 (1 H, dd, 3,
11, H17), 3.10 (1 H, m, H13), 5.54 (1 H, s, H18),
aꢀtetracarboxylic acid
H 7.42; N 6.05. C27H34N2O4 (
M
a
1
)
J
°С.
M
J
J
J
J
J
J
a
7.18 (1 H, s, H2'), 7.42 (1 H, s, H1'), 8.10 (1H, s, H3').
13C NMR spectrum: 14.1, 15.4, 15.6, 18.7, 19.2, 20.1,
25.7, 31.2, 33.4, 34.2, 35.3, 36.1, 36.6, 38.5, 44.3,
44.9, 47.7, 51.6, 51.8, 119.7 (C2'), 120.0 (C1'), 123.9
(C18), 133.5 (C3'), 146.5 (C19), 170.2 (C14), 171.8
(C16), 179.2 (C20).
J
12 Hz,
a
J
J
12, H2),
J
J12, H1), 3.57, 3.61,
3.63, and 3.75 (12 H, 4 s, 4OMe). 13C NMR spectrum:
13.7, 15.7, 17.4, 17.8, 18.0, 18.7, 21.9, 35.6, 35.9,
36.2, 36.8, 37.9, 43.9, 44.4, 46.9, 49.2, 50.4, 50.8,
50.9, 51.1, 51.2, 56.5, 59.1, 170.0 (COOCH3), 170.5
(COOCH3), 173.8 (COOCH3), 178.3 (COOCH3), 212.8
(C=O).
1ꢀ[(5R,9R,13R,17R)ꢀ5,9ꢀDimethylꢀ19ꢀ(1ꢀmethylꢀ
ethyl)ꢀ15ꢀoxaꢀ14,16ꢀdioxopentacyclo[10.5.2.01,10
.
04,9.013,17]heptadecꢀ18ꢀeneꢀ5ꢀcarbonyl]ꢀ4ꢀmethylpipꢀ
erazine (XIV). Yield 0.25 g (58%). Rf 0.78. Mp 161–
The acute toxicity of compounds (II), (III), and
(V)–(XV) was determined on 686 white nonꢀinbred
mice 18–22 g in mass of both sexes by introducing the
compounds in the stomach with a probe in doses from
2500 to 16000 mg/kg by the Kerber method [25]. The
animals were observed for 30 days. Based on the data
on the death of animals from various doses of the studꢀ
20
163°С. [α]D – 26
°
(с
0.05, CHCl3). Found, %: C 71.98;
497.67). Calculated, %:
C 72.17; H 8.77; N 5.80. H NMR spectrum: 0.59 (3
Н, s, СH3), 0.99 and 1.00 (6 H, both d, 6.8, 2СH3),
1.15 (3 H, s, CH3), 1.15–1.32 (1 H, m, H4), 1.35–1.90
H 8.62; N 5.64. C29H42N2O4 (
M
1
J
RUSSIAN JOURNAL OF BIOORGANIC CHEMISTRY Vol. 36
No. 6
2010

SYNTHESIS AND PHARMACOLOGICAL ACTIVITY OF AMIDES
769
ied compounds, the minimum lethal dose (LD100) and acute stomach ulcers. Indometacin and acetic ulcers
maximum endurable dose (LD0) were established by were induced by introducing a 3% solution of acetic
the integration method of Berens [26]. The LD16 and acid in a dose of 100 mg/kg (based on the active comꢀ
LD84 values were determined by the characteristic pound) or a solution of indometacin (20 mg/kg) into
curves built based on the integrated data. The variabilꢀ the rat’s stomach using a probe. The animals were kept
ity coefficient of the lethal doses (
by the equation
К
) was determined on a starvation diet. The compounds were introduced
as a water solution intragastrially 1 h before the introꢀ
duction of ulcerogens. The control group received disꢀ
tilled water. After 8 h, the ulcergogens were introduced
again. Then, the animals were kept for 24 h on a starꢀ
LD84
K = ꢀꢀꢀꢀꢀꢀꢀꢀꢀ.
LD16
vation diet at 4–6 С, after which a laparotomy was
°
The dose that led to the death of half of the animals,
LD50, was calculated by the Kerber equation [26]
conducted with the extraction of the stomachs. The
stomachs were opened along the greater curvature and
ulcers were counted twice using a magnifying glass.
The antiulcer activity (AA) was determined by the
equation
Σ(Zd)
LD50 = DM – ꢀꢀꢀꢀꢀꢀꢀꢀꢀꢀꢀꢀ ,
n
where DM is the dose that caused the death of all aniꢀ
AA = PI (control)/PI (experiment)
mals (LD100);
from two subsequent doses;
the numerical values of two adjacent doses;
Z
is half the number of animals that died
is the difference between
is the
is the number of animals in
Paules Index (PI) =
(А
×
В)/100,
d
where
А is the mean number of ulcers per animal and
∑
В
is the number of animals with ulcers in the group, %.
The results are given in Table 2.
summation sign; and
each group.
n
The mean error of LD50 was calculated by the Hadꢀ
dem equation
ACKNOWLEDGMENTS
This study was supported by the Russian Foundaꢀ
tion for Basic Research, project no. 09ꢀ03ꢀ00831.
k Sd
n
SLD50
=
ꢀꢀꢀꢀꢀꢀꢀꢀ ,
where
is the mean interval between the doses used;
number of animals in each group; and is the meanꢀ
square deviation LD50, which was calculated by the
equation
k
is the constant coefficient, which was 0.564;
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d
n
is the
S
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⎯
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