ISSN 10681620, Russian Journal of Bioorganic Chemistry, 2010, Vol. 36, No. 2, pp. 257–262. © Pleiades Publishing, Ltd., 2010. Original Russian Text © O.B. Kazakova, E.V. Tret’yakova, I.E. Smirnova, L.V. Spirikhin, G.A. Tolstikov, I.V. Chudov, G.V. Bazekin, A.F. Ismagilova, 2010, published in Bioorgan icheskaya Khimiya, 2010, Vol. 36, No. 2, pp. 277–282.
The Synthesis and AntiInflammatory Activity of Quinopimaric Acid Derivatives O. B. Kazakovaa,1, E. V. Tret’yakovaa, I. E. Smirnovaa, L. V. Spirikhina, G. A. Tolstikova, I. V. Chudovb, G. V. Bazekinb, and A. F. Ismagilovab a
Institute of Organic Chemistry, Ufa Research Centre, Russian Academy of Sciences, pr. Oktyabrya 71, Ufa, 450054 Russia b
Bashkir State Agricultural University, Russia
Received April 7, 2009; in final form, May 18, 2009
Under the action of PCl5, the Beckman rearrangement of a 3 : 1 mixture of Z and Eisomeres of 18βhydroxydihydroquinopimaric acid resulted in 5'caprolactam and isomeric caprolactams containing fragments of cyclic ether. Z and Eketoximes were separated as acetates. Using a carrageenan inflammation model, we demonstrated that the antiinflammatory activity of quinopimaric acid derivatives was comparable with that of diclofenac. Key words: antiinflammatory activity, Beckman rearrangement, diene adducts, lactams, levopimaric acid, oximes, quinopimaric acid, toxicity DOI: 10.1134/S1068162010020160
1
INTRODUCTION
The search for platforms for designing compounds with valuable pharmacological properties is one of the important tasks of modern medicinal chemistry [1]. Our studies are aimed at the synthetic transforma tions and activity studies of the diene adducts of levopimaric acid. Earlier, we showed that quinopi maric and 3chloroquinopimaric acid possess anti inflammatory activity [2] and dihydroquinopimaric and cyclopentenone pimaric acids displayed antiphlo gistic properties [3]. We also studied the antiviral activ ity of amides of dihydroquinopimaric acid towards the type A influenza virus [4]. RESULTS AND DISCUSSION In this work, we report the synthetic transforma tions of the dihydroquinopimaric acid E cycle and the preparation of its new nitrogencontaining derivatives, as well as the study of their antiinflammatory activity. The interaction of methyl 18βhydroquinopimaric 1 Corresponding
[email protected]
author; phone: +7 (347) 2356066; email:
acid (VI) with hydroxylamine hydrochloride in etha nol resulted in a ketoxime mixture (VII). In the 1H NMR spectrum, the resonance splitting of the meth oxy group and H20 was observed. Based on a compar ison of the integral intensities of these resonances, the ratio of isomeric ketoximes was found as a 3 : 1 mix ture. In the 13C NMR spectrum of crude ketoxime (VII), a double set of resonances of C5, C9, C10, C11, C12, C15, C16, C17, C18, C19, C20, and C21 was seen and the ratio of their integral intensities was also 3 : 1. Using the spectral characteristics of the Zketoxime of methyl dioxolanodihydroquinopimarate [5], we determined that the major reaction product was Zketoxime (VIIa) and the minor one was Eketoxime (VIIb). In the spectrum of Zketoxime (VIIa), the C15 re sonance was found at δ 159.7 ppm, whereas for Eketoxime it was located at δ 160.1 ppm. We sepa rated ketoximes (VIIa) and (VIIb) using their acetates (VIIIa) and (VIIIb). Homogeneous ketoxime (VIIa) was prepared by the alkaline treatment of acetate (VIIIa).
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O
OH
O 2 3
[2]
19
1 14
20 6 7 8
O COOH (I)
9
4
18 15
17 16
[8]
5 13 10 12 11 21
O COOH (II) R = H (III) R = CH3
OH (IV) R = COOCH3 (V) R = CH2OH
R
[8]
OH i
ii
O COOCH3
OAc
OAc
OH
+
(VI)
N
N
N COOCH3 OH (VII)
OR (VIIIa) (VIIIa) R = Ac (VIIa) R = H
COOCH3
RO (VIIIб) COOCH3 (VIIIb) R = Ac iii iii (VIIb) R = H
iv
OAc
OH
O
ii
NH
+
NH
O
COOCH3
O
NH
O
COOCH3 (XI)
+
NH O
O
COOCH3 (IX)
COOCH3 (Xa)
(Xb)
Conditions: i. NH2OH ⋅ HCl, EtOH, Δ; ii. Ac2O, DMAP, benzene, Δ; iii. Na2CO3, MeOH, Δ; iv. PCl5, Et2O. Scheme.
The Beckman rearrangement of crude ketoxime (VII) under the action of phosphorus pentachloride in diethyl ether yielded three products. The residue formed in the reaction corresponded to caprolactam (IX). Its 18βacetate (XI) was obtained in a yield of 90%. In the NMR spectra of the mixture, the reso nances of two other products can be seen. We assume that they belong to compounds (Xa) and (Xb): the presence of PCl5 supported the formation of an addi
tional cycle in ketoxime (VII) containing an ether C18–O–C19 bond. A similar effect was observed ear lier [6, 7]. Caprolactams (Xa) and (Xb) were isolated in yields of 5 and 24%, respectively, after the column chromatography of the mother solution. The study of the toxicological properties of com pounds (I)–(VI) showed that their toxicity varied from 4571.4 ± 303.6 for compound (IV) to 11178.6 ± 597.1 mg/kg for compound (V) (Table 1). Following the
Table 1. Acute toxicity of compounds (I)–(VI) Parameters of acute toxicity following the intragastric administration in mice Compound (I) (II) (III) (IV) (V) (VI)
LD0
LD16
LD50
LD84
LD100
K
5000 4000 4500 2500 7000 6500
6100 5900 5500 3050 8300 7300
8678.6 ± 494.4 8892.9 ± 575.3 7607.1 ± 393.8 4571.4 ± 303.6 11178.6 ± 597.1 10571.4 ± 636.3
11300 11700 9700 6100 14200 14000
12000 12500 10000 7000 16000 15500
1.85 1.98 1.76 2.00 1.71 1.92
Note: LD0 is the maximal dose not causing death; LD50, the mediumgrade lethal dose (a dose causing the death of 50% of experimental animals); LD100, the minimal dose causing death; and K, the variation coefficient of lethal doses. RUSSIAN JOURNAL OF BIOORGANIC CHEMISTRY
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Table 2. Antiinflammatory activity of compounds (I)–(VI), (VIIa), (VIIIa), and (IX) Doses and parameters of antiinflammatory activity 50 mg/kg
Compound
(I) (II) (III) (VI) (IV) (V) (VIIa) (VIIIa) (IX) Diclofenac (8 mg/kg) Control
100 mg/kg
% of growth of antiin flammatory edema
% of inhibition of anti inflammatory edema
% of growth of antiin flammatory edema
% of inhibition of anti inflammatory edema
39.21 ± 3.62 39.24 ± 2.24 39.96 ± 2.93 31.48 ± 2.52 41.36 ± 2.52 40.01 ± 1.97 35.77 ± 1.47 37.94 ± 1.96 36.26 ± 2.13 40.65 ± 1.54 54.42 ± 3.35
27.97 ± 2.58 27.91 ± 1.60 26.59 ± 1.95 42.17 ± 3.37 24.00 ± 2.52 26.50 ± 1.30 34.28 ± 1.40 30.30 ± 1.56 33.38 ± 1.96 25.31 ± 0.96 not studied
41.76 ± 3.75 39.65 ± 3.16 40.41 ± 3.75 33.68 ± 1.55 37.92 ± 2.52 40.40 ± 3.90 34.83 ± 1.96 37.52 ± 3.49 37.44 ± 2.10 40.65 ± 1.54 54.42 ± 3.35
23.27 ± 3.20 27.16 ± 2.85 25.75 ± 3.66 38.13 ± 1.76 30.33 ± 2.01 25.78 ± 2.49 36.00 ± 2.02 31.07 ± 3.72 31.11 ± 2.96 not studied not studied
administration in mice of compounds (I)–(VI) in doses close to LD16, a slight inhibition of motor activ ity (standing still) was observed in laboratory animals, but these signs disappeared as early as 1–2 h after the administration, followed by the complete recovery of motor activity. Following the administration of doses close to LD50 in mice, we marked an insignificant increase of motor activity for 20–30 min, followed by inhibition for 6–12 h. Absolutely lethal doses of diter penoids after intragastric administration caused rapid inhibition (for 5–10 min) followed by death in 12 (compound (IV)) to 48 h (compound (VI)). Hence, according to GOST (State Standard) 12.1.00.776, compounds (I)–(VI) were shown to belong to class 4 of danger (lowtoxic compounds). Variations of the toxic effects depended on the struc ture of quinopimaric acid derivatives. For example, the reduction of methyl dihydroquinopimarate to an 18βhydroxyl derivative (VI) resulted in a less toxic compound. On the contrary, the presence of two hydroxyl groups at positions C18 and C15 (compound (IV)) provided a considerable strengthening of toxicity (by a factor of two) as compared to quinopimaric acid (I). At the same time, the addition of a hydroxyimine fragment at the C15position did not change the toxic properties of the tested compounds. The studies of the antiinflammatory properties of the synthesized compounds (Table 2) showed that the effective dose of quinopimaric acid and its derivatives causing an antiinflammatory effect during carrag eenaninduced inflammation was about 50 mg/kg. In this dose, the antiinflammatory activities of quinopi maric acid (I) and its derivatives (I)–(VI), (VIIa), (VIIIa), and (IX) were comparable to that of diclofenac, widely used in veterinary and medicinal preparations, at the recommended dose of 8 mg/kg. RUSSIAN JOURNAL OF BIOORGANIC CHEMISTRY
The highest antiinflammatory activity was found for compound (VI); the inhibition index of the inflamma tory edema was 42.17 ± 3.37%, which is 16.86% higher than that of diclofenac. The antiinflammatory activ ities of compounds (VIIa), (VIIIa), and (IX) were more pronounced (by 5–9%) as compared to diclofenac, but were reliably lower as compared to compound (VI) (by 9–11%). These figures demonstrate that under the given laboratory model, the presence of a hydroxy imine group at the C15position of the tested com pounds provided for an invariance of toxicity and a decrease in the antiinflammatory activity. An increase in the dose of the tested compounds up to 100 mg/kg did not cause a rise in their antiinflam matory activity. Only for compound (IV) was the effect more pronounced (by 6.33%) as compared to the dose of 50 mg/kg, but it was reliably lower (by 11.90%) as compared to the same dose of compound (IV). The performed studies showed that a substituent at the C15position considerably affected the toxic effect and the pharmacological activity of the tested com pounds. Thus, using a carrageenan model, we found that quinopimaric acid derivatives (IV), (VI), (VIIa), (VIIIa), and (IX) possessed higher antiinflammatory activities than diclofenac. EXPERIMENTAL 1H
and 13C NMR spectra were registered on a Bruker AM300 spectrometer (Germany) with a working frequency of 300 and 75.5 MHz, respectively; (δ, ppm, J, Hz) in CDCl3; the internal standard was tetramethylsilane. The melting points were measured on a Boetius microtable. The optical rotation was measured on a PerkinElmer 241 MC polarimeter (Germany) in a tube 1 dm in length. TLC was per Vol. 36
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formed on Silufol plates (Chemapol, Czech Republic) in a 20 : 1 mixture of chloroform–methanol. The compounds were developed with 5% phosphotungstic acid in ethanol followed by heating at 100–120°C for 2–3 min. Compounds (II) and (IV)–(VI) were obtained according to [2, 8]. 18Hydroxy5,9dimethyl9methoxycarbonyl 19(1methylethyl)15hydroxyiminopentacyclo [12.5.0.22,13.04,13.05,10]eicos20ene (VII). A solution of compound (VI) (1 mmol, 0.42 g) and hydroxy lamine hydrochloride (0.5 g) in ethanol (15 ml) was refluxed for 8 h. The mixture was poured into 5% HCl (30 ml), the residue was filtered, washed with water, dried in air, and recrystallized from acetone to give 0.38 g (90%) of the product; Rf 0.61; mp 125–127°C. 1H NMR: 0.60 (3 H, s, CH ), 0.82–1.00 (2 H, m, CH), 3 1.05 and 1.08 (6 H, both d, J 6.9, 2CH3), 1.15 (3 H, s, CH3), 1.27–1.79 (12 H, m, CH, CH2), 2.10–2.50 (8 H, m, CH, CH2), 2.65 (1 H, br s, H2), 3.40 (1 H, br s, OH), 3.65 (0.7 × 3H, s, OCH3), 3.66 (0.3 × 3 H, s, OCH3), 5.43 (0.7 × 1 H, br s, H20), 5.46 (0.3 × 1 H, br s, H20), 9.46 (1 H, br s, NOH). 13C NMR: (179.5, 179.4) (C21), (160.1, 159.7) (C15), (147.6, 146.6) (C19), (124.5, 124.0) (C20), (68.9, 68.1) (C18), 55.8, 53.9, 51.9, 49.6, 47.3, (47.2, 46.6), (45.9, 44.9), (41.2, 40.8), 38.2, 37.9, (36.9, 36.8), 35.7, 33.9, (33.6, 33.5), (33.1, 33.0), 29.7, 25.8, 21.9, 19.4, 19.1, 17.1, 16.6, 15.9. Compounds (VIIIa) and (VIIIb). Acetic anhydride (0.2 ml) and dimethylaminopyridine (0.25 g) were added to a solution of compound (VII) (1 mmol, 0.44 g) in dry benzene (20 ml) and the mixture was stirred for 18 h at room temperature. The mixture was poured in 5% HCl (30 ml), washed with water (2 × 100 ml), and dried with CaCl2. The solvent was evap orated in a vacuum, and the residue was purified by column chromatography on Al2O3 with chloroform as the eluent. (15Z)18Acetoxy5,9dimethyl9methoxycar bonyl19(1methylethyl)15hydroxyiminopentacy clo[12.4.0.22,13.04,13.05,10]eicos20ene (VIIIa). Yield 0.31 g (76%). Rf 0.58; mp 191–193°С. [ α ]20 D +34.0° (с 0.005, CHCl3). Found, %: C 70.35, H 8.55, N 2.21. C32H47NO6. Calc., %: C 70.95, H 8.74, N 2.59. 1H NMR: 0.49 (3 Н, s, CH3), 0.72–0.90 (2 H, m, CH), 1.00 and 1.02 (6 H, both d, J 6.9, 2СН3), 1.05 (3 Н, s, СН3), 1.27–1.79 (11 H, m, CH, СН2), 1.98 (3 Н, s, OCH3), 2.02 (3 H, s, NOAc), 2.10–2.50 (10 H, m, CH, CH2), 2.65 (1 H, br s, H2), 3.55 (3 H, s, OCH3), 4.51 (1 H, dt, J1 4.9, J2 6.4, J3 10.7, H18), 5.33 (1 H, br s, H20). 13C NMR: 178.7 (C21), 169.7 (OC–O), 168.4 (OC–O), 166.4 (C15), 147.6 (C19), 123.8 (C20), 70.3 (C18), 55.5, 52.9, 51.7, 49.4, 48.9, 46.9, 42.5, 40.3, 37.8, 37.6, 36.3, 34.6, 34.2, 34.0, 32.7, 28.9, 26.2, 22.3, 21.4, 20.9,19.4, 19.3, 16.8, 16.5, 15.6. (15E)18Acetoxy5,9dimethyl9methoxycarbo nyl19(1methylethyl)15hydroxyiminopentacy
clo[12.4.0.22,13.04,13.05,10]eicos20ene (VIIIb). Yield 0.06 g (15%). Rf 0.60; mp 178–180°С. [ α ]20 D +14.0° (с 0.005, CHCl3). Found, %: C 70.35, H 8.87, N 2.31. C32H47NO6. Calc., %: C 70.95, H 8.74, N 2.59. 1H NMR: 0.50 (3 Н, s, СН3), 0.72–0.90 (2 H, m, CH), 1.01 and 1.03 (6 H, both d, J 6.9, 2СН3), 1.06 (3 Н, s, СН3), 1.27–1.80 (11 H, m, CH, СН2), 1.98 (3 Н, s, OCH3), 2.01 (3 H, s, NOAc), 2.10–2.50 (10 H, m, CH, CH2), 2.65 (1 H, br s, H2), 3.55 (3 H, s, OCH3), 4.45 (1 H, dt, J1 4.9, J2 6.4, J3 10.7, H18), 5.30 (1 H, br s, H20). 13C NMR: 178.5 (C21), 169.5 (OC–O), 168.1 (OC–O), 166.4 (C15), 146.6 (C19), 123.6 (C20), 69.7 (C18), 55.3, 52.8, 51.7, 49.3, 48.9, 46.9, 42.0, 40.3, 37.6, 37.4, 36.3, 34.4, 34.1, 34.0, 32.7, 28.9, 26.1, 22.3, 21.1, 20.9, 19.4, 19.3,16.8, 16.5, 15.5. (15Z)18Hydroxy5,9dimethyl9methoxycarbo nyl19(1methylethyl)15hydroxyiminopentacyclo [12.4.0.22,13.04,13.05,10]eicos20ene (VIIa). Sodium carbonate (4.1 mmol, 0.43 g) was added to a solution of compound (VIIIa) (1 mmol, 0.54 g) in methanol (30 ml) and the mixture was refluxed for 4 h. The solu tion was poured into 5% HCl (30 ml); the residue was filtered, washed with water, and dried in air to give 0.50 g (93%) of the product. Rf 0.61; mp 135–138°С. [ α ]20 D +14.0° (с 0.005, CHCl3). Found, %: C 72.90, H 8.42, N 3.10. C27H41NO4. Calc., %: C 73.10, H 8.32, N 3.16. 1H NMR: 0.60 (3 Н, s, СН3), 0.82–1.00 (2 H, m, CH), 1.05 and 1.08 (6 H, both d, J 6.9, 2 CH3), 1.15 (3 H, s, CH3), 1.27–1.79 (12 H, m, CH, CH2), 2.10– 2.50 (8 H, m, CH, CH2), 2.65 (1 H, br s, H2), 3.40 (1 H, br s, OH), 3.65 (3 H, s, OCH3), 5.43 (1 H, br s, H20), 9.46 (1 H, br s, NOH). 13C NMR: 179.4 (C21), 159.7 (C15), 147.8 (C19), 124.0 (C20), 68.9 (C18), 55.8, 53.9, 51.9, 49.6, 47.3, 47.2, 45.9, 41.2, 38.2, 37.9, 36.8, 35.7, 33.9, 33.6, 33.1, 29.7, 25.8, 21.9, 19.4, 19.1, 17.1, 16.6, 15.9. Synthesis of compounds (IX), (Xa), and (Xb). Phos phorus pentachloride (4 mmol, 0.82 g) was added to a solution of compound (VII) (2 mmol, 0.44 g) in diethyl ether (50 ml) and the mixture was stirred for 30 min. The precipitate was filtered, washed with cool ether, and dried to give 0.22 g (50%) of compound (IX). The mother solution was poured into 5% HCl (30 ml), washed with water (2 × 100 ml), and dried with CaCl2. The solvent was evaporated in a vacuum and the residue was purified by column chromatography on Al2O3 eluted with chloroform to give 0.02 g (5%) of compound (Xa) and 0.11 g (24%) of compound (Xb). 18Hydroxy5,9dimethyl9methoxycarbonyl19 (1methylethyl)15azaheptacyclo[12.5.0.22,13.04,13.05,10] geneicos20en16one (IX). Rf 0.35; mp 166–168°С. [ α ]20 D +6.0° (с 0.005, CHCl3). Found, %: C 72.97, H 9.22, N 3.18. C27H41NO4. Calc., %: C 73.10, H 9.32, N 3.16. 1H NMR: 0.50 (3 Н, s, СН3), 0.82–1.00 (2 H, m, CH), 1.05 and 1.08 (6 H, J 6.9, both d, 2СН3), 1.15
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(3 Н, s, СН3), 1.27–1.79 (12 H, m, CH, СН2), 2.10– 2.50 (8 H, m, CH, СН2), 3.20 (1 H, br s, H2), 3.65 (3 Н, s, ОСН3), 4.50 (1 H, br s, OH), 5.20 (1 H, br s, H20), 7.66 (1 H, br s, NH). 13C NMR: 178.4 (C21), 176.3 (C15), 148.4 (C19), 122.9 (C20), 79.4 (C18), 59.6, 52.0, 50.6, 48.8, 47.6, 46.6, 40.7, 40.3, 39.7, 39.4, 38.8, 36.9, 36.4, 33.6, 32.5, 28.1, 27.7, 20.7, 19.8, 16.8, 16.6, 15.5. 5,9Dimethyl9methoxycarbonyl19(1methylethyl) 18oxa14azaoctacyclo[12.5.0.22,13.04,13.05,10.119.21] docosan15one (Xa). Rf 0.48; mp 129–131°С. [ α ]20 D +8.0° (с 0.005, CHCl 3). Found, %: C 72.75, H 9.15, N 3.28. C27H41NO4. Calc., %: C 73.10, H 9.32, N 3.16. 1H NMR: 0.86 (3 Н, s, СН3), 0.89 and 0.93 (6 H, both d, J 7.11, 2СН3), 1.06 (3 Н, s, СН3), 1.09–1.19 (2 H, m, CH), 1.23–1.80 (11 H, m, CH, СН2), 1.96–2.24 (6 H, m, CH, СН2), 2.37–2.57 (5 H, m, CH, СН2), 3.57 (3 Н, s, СН3), 4.10 (1 H, br s, H18), 6.10 (1 H, br s, NH). 13C NMR: 178.9 (C21), 178.4 (C15), 83.2 (C19), 76.6 (C18), 65.2, 55.3, 51.9, 51.8, 49.9, 47.9, 47.1, 40.0, 38.7, 38.6, 37.8, 36.4, 36.0, 35.8, 34.8, 34.1, 31.9, 21.1, 17.1, 16.9, 16.7, 16.5, 15.5. 5,9Dimethyl9methoxycarbonyl19(1methylethyl) 18oxa16azaoctacyclo[12.5.0.22,13.04,13.05,10.119.21] geneicos15one (Xb). Rf 0.48; mp 115–117°С. [ α ]20 D +12.0° (с 0.005, CHCl 3). Found, %: C 72.85, H 9.12, N 3.21. C27H41NO4. Calc., %: C 73.10, H 9.32, N 3.16. 1H NMR: 0.86 (3 Н, s, СН3), 0.89 and 0.93 (6 H, both d, J 7.11, 2СН3), 1.06 (3 Н, s, СН3), 1 (2 H, m, CH), 1.23–1.80 (11 H, m, CH, СН2), 1.96–2.24 (6 H, m, CH, CH2), 2.37–2.57 (5 H, m, CH, СН2), 3.57 (3 H, s, CH3), 3.85 (1 H, br s, H18), 6.69 (1 H, br s, NH). 13C NMR: 178.9 (C21), 176.8 (C15), 84.3 (C19), 74.8 (C18), 65.2, 55.3, 51.9, 51.8, 49.9, 47.9, 47.1, 40.0, 38.7, 38.6, 37.8, 36.4, 36.0, 35.8, 34.8, 34.1, 31.9, 21.1, 17.1, 16.9, 16.7, 6.5, 15.5. 18Acetoxy5,9dimethyl9methoxycarbonyl19 (1methylethyl)15azaheptacyclo[12.5.0.22,13.04,13.05,10] geneicos20en15one (XI). Acetic anhydride (0.2 ml) and dimethylaminopyridine (0.25 g) were added to a solution of compound (IX) (1 mmol, 0.44 g) in dry benzene (20 ml) and refluxed for 8 h. The mixture was poured into 5% HCl (30 ml), washed with water (2 × 100 ml), and dried with CaCl2. The solvent was evaporated in a vacuum, and the residue was puri fied by column chromatography over Al2O3 eluted with chloroform. Yield, 0.39 g (90%). Rf 0.58; mp 200– 202°С. [ α ]20 D +44.0° (с 0.005, CHCl3). Found, %: 71.70, H 8.78, N 2.60. C29H43NO5. Calc., %: C 71.72, H 8.92, N 2.88. 1H NMR: 0.49 (3 Н, s, СН3), 0.72–0.90 (2 H, m, CH), 1.00 and 1.02 (6 H, both d, J 6.9, 2СН3), 1.05 (3 Н, s, СН3), 1.27–1.65 (12 H, m, CH, СН2), 1.89 RUSSIAN JOURNAL OF BIOORGANIC CHEMISTRY
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(3 H, s, OAc), 1.98–2.45 (11 H, m, CH, CH2), 3.51 (3 H, s, OCH3), 4.21 (1 H, dt, J1 4.9, J2 6.4, J3 10.7, H18), 5.25 (1 H, d, J 10.5, NH), 5.43 (1 H, br s, H20). 13C NMR: 178.9 (C21), 176.9 (C15), 169.7 (OCO), 150.4 (C 19), 123.8 (C20), 81.3 (C18), 57.3, 51.9, 51.7, 49.4, 49.0, 48.3, 47.0, 42.0, 38.2, 37.6, 37.4, 36.4, 33.6, 32.9, 30.4, 28.4, 23.3, 21.6, 21.0, 20.2, 17.0, 16.5, 15.5. The acute toxicity of the tested compounds was studied on 605 masscalibrated white mice (body weight of 18–20 g) preliminarily subjected to a 15day quarantine according to the protocol described in [9]. Compounds (I)–(VI) were administered intragastri cally in doses of 2500 to 16000 mg/kg of body weight. The animals were observed for 30 days. Based on the death of the animal following the administration of varied doses of the tested compounds, the absolutely lethal dose (LD100) and a maximally tolerated dose (LD0) were found by integration using the Berenice method [9, 10]. The LD16 and LD84 values found by the characteristic curves plotted based on the integra tion data were used for the determination of the varia tion coefficients of lethal doses [11]. The antiinflammatory properties of the com pounds were studied using a carrageenan inflamma tion model [12]. The antiinflammatory effect was estimated by volume variations of mouse pads affected by flagogen. The effect was calculated by the following formula: V −V inflammation inhibition (%) = k 0 , Vk where Vk is the mean increase in the mouse pad vol ume in the control group, and V0 is the mean increase in the mouse pad volume in the experimental group. The reproduction of experimental pathologies was conducted in animals anaesthetized with sodium ethaminal. Decapitation was carried out using ether anesthesia. REFERENCES 1. Newman, D.J. and Cragg, G.M., J. Nat. Prod., 2007, vol. 70, pp. 461–477. 2. Flekhter, O.B., Tret’yakova, E.V., Galin, F.Z., Kara churina, L.T., Spirikhin, L.V., Zarudii, F.S., and Tolstikov, G.A., Khim.Farm. Zh., 2002, vol. 36, pp. 29–31. 3. Flekhter, O.B., Tret’yakova, E.V., Makara, N.S., Gab drakhmanova, S.F., Baschenko, N.Zh., Galin, F.Z., Zarudii, F.S., and Tolstikov, G.A., Khim.Farm. Zh., 2003, vol. 37, pp. 35–37. 4. Flekhter, O.B., Smirnova, I.E., Tret’yakova, E.V., Tol stikov, G.A., Savinova, O.V., and Boreko, E.I., Bioorg. Khim., 2009, vol. 35, pp. 424–430 [Russ. J. Bioorg. Chem. (Engl. Transl.), 2009, vol. 35, pp. 385–390]. 5. Smirnova, I.E., Tret’yakova, E.V., Kazakova, O.B., and Starikova, Z.A., Zh. Strukt. Khim., 2009, pp. 379⎯381. 6. Herz, W., Blakstone, R.C., and Nair, M.G., J. Org. Chem., 1967, vol. 32, pp. 2292–2998. Vol. 36
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7. Smirnova, I.E., Tret’yakova, E.V., Kazakova, O.B., Starikova, Z.A., and Fedyanin, I.V., Zh. Strukt. Khim., 2009, pp. 377–378. 8. Smirnova, I.E., Tret’yakova, E.V., Flekhter, O.B., Spir ikhin, L.V., Galin, F.Z., Tolstikov, G.A., Starikova, Z.A., and Korlyukov, A.A., Zh. Org. Khim., 2008, vol. 44, pp. 1623–1629. 9. Sanotskii, I.V., Metody opredeleniya toksichnosti i opas nosti khimicheskikh veshchestv (Methods of Determina tion of Toxicity and Hazard of Chemicals), Moscow: Meditsina, 1970.
10. Elizarova, O.N., Opredelenie porogovykh doz promy shlennykh yadov pri peroral’nom vvedenii (Determina tion of Threshold Doses of Industrial Poisons at Peroral Administration), Moscow: Meditsina, 1971. 11. Belen’kii, M.P., Osnovnye priemy statisticheskoi obrabo tki rezul’tatov nablyudenii v oblasti fiziologii (The Main Methods of Statistical Processing of Results of Obser vations in Physiology), Moscow: Meditsina, 1963. 12. Trinus, F.P. and Mokhort, N.F., Nesteroidnye protivo vospalitel’nye sredstva (Nonsteroid Antiinflammatory Drugs), Kiev, 1975.
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