Synthesis of a Macrocyclic Conjugate of the Diterpenoid Isosteviol and Glucuronic Acid

Article abstract of DOI:10.1007/s10600-015-1385-6

A macrocyclic conjugate of the natural diterpenoid isosteviol (16-oxo-ent-beyeran-19-oic acid) and glucuronic acid was synthesized for the first time. The conjugate contained two molecules of dihydroisosteviol β-D-glucuronoside joined by 1,8-octanedicarboxylate and 1,8-octanedicarbazoyl spacers.

Full text of DOI:10.1007/s10600-015-1385-6

DOI 10.1007/s10600-015-1385-6
Chemistry of Natural Compounds, Vol. 51, No. 4, July, 2015
SYNTHESIS OF A MACROCYCLIC CONJUGATE OF
THE DITERPENOID ISOSTEVIOL AND GLUCURONIC ACID
O. V. Andreeva, R. R. Sharipova, B. F. Garifullin,
I. Yu. Strobykina, and V. E. Kataev*
A macrocyclic conjugate of the natural diterpenoid isosteviol (16-oxo-ent-beyeran-19-oic acid) and glucuronic
acid was synthesized for the first time. The conjugate contained two molecules of dihydroisosteviol
ꢀ
-D-glucuronoside joined by 1,8-octanedicarboxylate and 1,8-octanedicarbazoyl spacers.
Keywords: isosteviol, glucuronic acid, glycophanes, macrocycles, macrocyclic glycoterpenoids.
We recently reported the synthesis of the first conjugate of the diterpenoid isosteviol (16-oxo-ent-beyeran-19-oic
acid) and glucuronic acid [1] in which two molecules of the isosteviol derivative dihydroisosteviol (16-hydroxy-ent-beyeran-
1
9-oic acid) functionalized at the carboxyls by ꢀ-D-glucopyranuronoyl residues were connected by a 1,6-hexanedicarboxylate
spacer. In continuation of research on the synthesis of macrocyclic glycoterpenoids [2], herein we present results for the
synthesis of a macrocyclic conjugate of isosteviol and glucuronic acid. We used the methodology for synthesizing macrocyclic
isosteviol derivatives [3, 4] in which two dihydroisosteviol molecules were first bound through the hydroxyls by reacting them
with carboxylic acid chlorides. The resulting diacids (e.g., diacid 4 in Scheme 1) were converted to macrocycles by reacting
their carboxylic acid chlorides with diols.
The starting carbohydrate component was D-(+)-glucurono-3,6-lactone (1), which was easily converted to methyl-
1
,2,3,4-tetra-O-acetyl-ꢀ-D-glucopyranuronate (2) as before [5]. The methyl-1-deoxy-1-bromo-2,3,4-tri-O-acetyl-ꢀ-D-
glucopyranuronate 3 that was obtained from it by the literature method [6] was then reacted with diacid 4 in refluxing CH Cl
2
2
in the presence of tetrabutylammonium bromide (TBAB) and potash. Bisglucuronide 5 was obtained in 56% yield. Its
anomeric protons appeared in the PMR spectrum as a single doublet at 5.73 ppm with vicinal SSCC 8.02 Hz. This indicated
convincingly that the glycoside bonds had the ꢀ-orientation.
In fact, the anomeric proton of methyl 1,2,3,4-tetra-O-acetyl-ꢀ-D-glucopyranuronate 2 resonated as a doublet at
5
.73 ppm with vicinal SSCC 7.8 Hz [6] whereas the anomeric proton of the corresponding ꢁ-D-glucopyranuronate was found
as a doublet at 6.34 ppm with SSCC 3.7 Hz [5]. Then, bisglucuronoside 5 was reacted at room temperature with hydrazine
hydrate as before [7]. The PMR spectrum of the product, which was obtained in 90% yield, lacked the strong singlets
corresponding to acetate resonances at 1.96, 1.98, and 1.99 ppm and the ester methyls of the glucopyranuronates at 3.68 ppm,
which were observed in the spectrum of 5. A multiplet that was characteristic of the hydrazide NH proton appeared at
7
.69–7.73 ppm. All these data combined with MALDI mass spectral data indicated that dihydrazide 6 had formed.
The reaction did not affect the carbohydrate anomeric centers. Therefore, the glycoside bonds in 6 retained the ꢀ-orientation.
Its glucuronate anomeric protons resonated as a single doublet at 3.78 ppm with SSCC 9.63 Hz, which agreed with
3
the literature for various ꢀ-glycosides of non-acylated glucuronic acid methyl ester (e.g., ꢂ 4.26 ppm and J = 9.4 Hz [8]) and
with previous work [9] in which the PMR spectrum of 1-O-ꢁ-acetyl-D-glucuronic acid was described. The anomeric proton
in this compound occupied the ꢀ-position and still resonated at 5.02 ppm as a doublet with a much smaller constant of 4.2 Hz.
The macrocycle was formed by reacting dihydrazide 6 with sebacic acid dichloride. Bisglucuronoside 6 was thoroughly
–
1
dehydrated before use. The reaction was carried out for 72 h until the ꢃ(C=O) stretching band at 1800 cm that was characteristic
of sebacic acid dichloride vibrations disappeared in the IR spectrum of the reaction mixture.
A. E. Arbuzov Institute of Organic and Physical Chemistry, Kazan Scientific Center, Russian Academy of Sciences,
420088, Kazan, Ul. Arbuzova, 8, fax: (843) 273 22 53, e-mail: kataev@iopc.ru. Translated from Khimiya Prirodnykh Soedinenii,
No. 4, July–August, 2015, pp. 594–597. Original article submitted February 27, 2015.
0
009-3130/15/5104-0689 ©2015 Springer Science+Business Media New York
689

OMe
OMe
HO
O
OAc
O
Br
O
i, ii
O
O
iii
O
OH
O
AcO
OAc
AcO
OAc
OH
OAc
OAc
1
2
3
O
O
O
O
O
O
4
1
7
1
6
O
O
4
13
15
1
1
9
7
20
iv
v
3
+
1
5
18
3
19
O
O
O
O
HO
O
O
OH
OAc
AcO
AcO
4
O
1'
O
3'
MeO
OMe
5'
OAc
O
OAc
OAc O
5
O
O
O
O
O
O
4
O
O
4
vi
O
O
O
O
O
O
O
O
O
OH
OH
HO
HO
OH
OH
HO
HO
O
O
O
O
H
H
N
O
O
H2N
N
NH2
O
O
OH
N
H
OH
NH
OH
OH
O
N
H
N
H
4
6
7
i. MeONa, CH OH, room temp., yield 100%; ii. Ac O, HClO , 0°C, yield 50%; iii. 33% HBr–AcOH, CH Cl , 0°C, yield 100%; iv. TBAB,
3
2
4
2
2
K CO , CH Cl –H O, reflux, yield 56%; v. MeOH–CHCl , NH NH 4H O, room temp., yield 90%; vi. CHCl –C H N, ClOC(CH ) COCl,
2
3
2
2
2
3
2
2
2
3
5
5
2 8
ꢄ
4
A mol. sieves, room temp., yield 30%
The reaction product that was isolated by column chromatography over silica gel in 30% yield was macrocycle 7
according to MALDI mass spectrometry. The PMR spectrum of the macrocycle contained the same characteristic isosteviol
methyl resonances as dihydrazide 6. Resonances of the 1,8-octanedicarboxylate and 1,8-octanedicarbazoyl spacers overlapped
each other in the spectrum. The number of protons was determined from the integrated intensities. The dihydrazide protons
resonated as multiplets at 7.31–7.38 and 7.71–7.77 ppm. Like for macrocyclic isosteviol derivatives with the same fragments
[
3], this was apparently indicative of several hindered conformers of macrocycle 7. The anomeric protons of its glucuronates
were found in the spectrum as a single doublet at 3.88 ppm with SSCC 9.51 Hz. The resonances of the carbohydrate anomeric
protons in 5–7 as single doublets with SSCC 8–9.6 Hz provided unambiguous evidence that they all were ꢀ-glucuronosides.
Several macrocyclic glucuronic acid derivatives in which carbohydrate residues were connected by polymethylene
[
10] or alkenylene [11] spacers were reported. However, macrocycles containing terpenoid fragments in addition to glucuronic
acid have not been reported. Thus, it could be concluded that we synthesized for the first time macrocyclic glycoterpenoids as
the macrocyclic conjugate of the diterpenoid isosteviol and glucuronic acid.
690

EXPERIMENTAL
PMR spectra were recorded on Avance-400 andAvance-500 spectrometers (Bruker, Germany). MALDI mass spectra
were obtained in an UltraFlex III TOF/TOF time-of-flight mass spectrometer (Bruker Daltonik GmbH, Germany). Measurements
were made in the range m/z 400–3000. The data were processed using the FlexAnalysis 3.0 programs (Bruker Daltonik
GmbH, Germany). The matrix consisted of p-nitroaniline. Samples were dissolved in CHCl or CHCl –MeOH (1:1) at a
3
3
–
3
concentration of 10 mg/mL and deposited by the dried drop method. Optical rotation was measured on a PerkinElmer 341
polarimeter (USA). IR spectra were recorded from films on a Vector 22 Fourier spectrometer (Bruker, Germany) in the range
00–4000 cm . Melting points were determined on a Boetius apparatus. The course of reactions and purity of products were
–
1
4
monitored by TLC on Sorbfil PTSKh-AF-Aplate (Krasnodar, Russia) with detection by H SO (5%) and heating. Compounds
2
4
were isolated by flash chromatography over KSK silica gel (0.063–0.125 mm fraction, OOO KhromLab, Russia).
2
0
Compound 2 was prepared by the literature method [5], mp 177°C (MeOH), [ꢁ] +8.1° (c 1.1, CHCl ); lit. mp
D
3
5
1
76°C [5], mp 177–178°C (EtOH), [ꢁ]2 +7.4° (c 2.0, CHCl ) [12]. Compound 3 was prepared by the literature method [6],
D 3
mp 105–106°C (EtOAc); lit. mp 106–107°C (EtOH) [12]. Diacid 4 was prepared by the literature method [13], mp 108–
2
0
20
1
09°C, [ꢁ] –97.7° (c 1.1, CHCl ); lit. mp 108–110°C, [ꢁ] –99° (c 0.17, C H ) [13]. We used commercial D-(+)-glucurono-
D
3
D
6 6
6
,3-lactone (Acros, Belgium).
Bis[19-nor-4ꢁ(2,3,4-tetra-O-acetyl-5-methoxycarbonyl-ꢀ-D-glucopyranosyloxycarbonyl)-ent-beyeran-16-yl]-
,8-octanedicarboxylate (5). Bromide 3 (0.5 g, 1.26 mmol) and diacid 4 (0.51 g, 0.63 mmol) were dissolved in freshly
1
distilled CH Cl (50 mL), treated with K CO (0.55 g, 4 mmol) and TBAB (0.4 g, 1.26 mmol), diluted with H O (5 mL),
2
2
2
3
2
stirred for 1 h at room temperature, and refluxed for 16 h. The solution became dark during the reaction. The reaction mixture
was cooled, diluted with CHCl , washed with H O, and dried over MgSO . The solvent was distilled at reduced pressure.
3
2
4
Chromatography of the residue over silica gel (petroleum ether–EtOAc eluent, 2:1) afforded a product that was dried in vacuo
2
0
–1
to give a white powder. Yield 0.5 g (56%), mp 114–115ꢄÑ, [ꢁ] –39.9ꢄ (ñ 1.1, CHCl ). IR spectrum (ꢃ, cm ): 1759 (C=O).
D
3
1
Í NMR spectrum (400 MHz, ÑDÑl , ꢂ, ppm, J/Hz): 0.70–1.90 (50Í, m, ent-beyerane skeleton, spacer fragment (ÑÍ ) ),
3
2 6
0
.66 (6Í, s, H -20), 0.87 (6Í, s, H -17), 1.16 (6Í, s, Í -18), 1.96 (6H, s, CH C(O)), 1.98 (6H, s, CH C(O)), 1.99 (6H, s,
3 3 3 3 3
CH C(O)), 2.11 (2H, d, J = 13.08, H -3), 2.25 (4H, t, J = 7.43, two “upper” spacer fragments 16-OC(O)CH ), 3.68 (6H, s,
3
eq
2
H -6ꢅ), 4.06–4.14 (2H, m, H-5ꢅ), 4.66 (2Í, dd, J = 10.7, 4.07, H-16), 5.14–5.29 (6H, m, H-2ꢅ, 3ꢅ, 4ꢅ), 5.73 (2H, d, J = 8.02,
3
+
+
H-1ꢅ). Mass spectrum: m/z 1461.8 [M + Na] , calcd 1461.7 [M + Na] . C H O .
7
6 110 26
Bis[19-nor-4ꢁ(5-carbazoyl-ꢀ-D-glucopyranosyloxycarbonyl)-ent-beyeran-16-yl]-1,8-octanedicarboxylate (6).
Bis-derivative 5 (0.5 g, 0.35 mmol) was dissolved in a mixture of MeOH (30 mL) and CHCl (10 mL), treated with hydrazine
3
hydrate (0.5 mL), and held at room temperature until a crystalline precipitate formed. The obtained product was rinsed with
2
0
–1
MeOH. Yield 0.37 g (90%), mp 131–132ꢄÑ, [ꢁ] –41.2ꢄ (ñ 0.7, CHCl –MeOH 1:1). IR spectrum (ꢃ, cm ): 1373, 1610,
D
3
1
1
1
2
3
674 (O=C–NH), 1759 (C=O), 3323 (OH, NH, NH ). Í NMR spectrum (400 MHz, ÑDÑl –CD OD 1:1, ꢂ, ppm, J/Hz): 0.80–
2 3 3
.90 (50Í, m, ent-beyerane skeleton, spacer fragment (ÑÍ ) ), 0.77 (6Í, s, H -20), 0.89 (6Í, s, H -17), 1.19 (6Í, s, Í -18),
.16 (2H, d, J = 13.30, H -3), 2.30 (4H, t, J = 7.38, two “upper” spacer fragments 16-OC(O)CH ), 3.31–3.35 (2H, m, H-2ꢅ),
.39–3.46 (4H, m, H-3ꢅ, 4ꢅ), 3.55–3.61 (2H, m, H-5ꢅ), 3.78 (2H, d, J = 9.63, H-1ꢅ), 4.67 (2Í, dd, J = 10.6, 4.2, H-16), 5.39–5.42
2H, m, 2 ꢆ OH), 7.69–7.73 (2H, m, NH). Mass spectrum: m/z 1209.5 [M + Na] , calcd 1209.7 [M + Na] . C H N O .
2
6
3
3
3
eq
2
+
+
(
62 98 4 18
2
,13,16,35-Tetraoxa-19,20,31,32-tetraaza-1,14(16,4ꢁ)di(19-nor-ent-beyerane)-17,34(1,5)di(ꢀ-D-
glucopyranosyl)cyclohexatriacontaphane-3,12,15,18,21,30,33,36-octaone (7). Dihydrazide 6 (0.3 g, 0.25 mmol) was stored
°
in vacuo for 3 h, dissolved in a mixture of anhydrous CHCl (50 mL) and freshly distilled Py (5 mL), treated with 4A molecular
3
sieves, stored for 1 h, treated with sebacic acid chloride (0.06 g, 0.25 mmol), and stirred at room temperature for 72 h.
The solvent was removed at reduced pressure. Chromatography of the residue over silica gel (EtOAc–MeOH eluent, 4:1)
2
0
–1
afforded a white amorphous powder. Yield 0.1 g (30%), mp 176–178ꢄÑ, [ꢁ] –34.9ꢄ (ñ 0.55, MeOH). IR spectrum (ꢃ, cm ):
D
1
1
372, 1630, 1670 (O=C–NH), 1730 (C=O), 3290 (OH, NH). Í NMR spectrum (400 MHz, ÑDÑl –CD OD 1:1, ꢂ, ppm,
3
3
J/Hz): 0.60–1.80 (62Í, m, ent-beyerane skeleton, “upper” spacer fragment (ÑÍ ) and “lower” spacer fragment (ÑÍ ) ),
2
6
2 6
0
.67 (6Í, s, H -20), 0.79 (6Í, s, H -17), 1.11 (6Í, s, Í -18), 2.08 (2H, d, J = 12.82, H -3), 2.11–2.25 (8H, m, two “upper”
3 3 3 eq
spacer fragments 16-OC(O)CH and two “lower” spacer fragments NC(O)CH ), 3.38–3.48 (4H, m, H-2ꢅ, 3ꢅ), 3.51–3.62 (4H,
2
2
m, H-4ꢅ, 5ꢅ), 3.88 (2H, d, J = 9.51, H-1ꢅ), 4.58 (2Í, dd, J = 10.2, 4.7, H-16), 5.38 (2H, d, J = 7.48, 2 ꢆ OH), 7.31–7.38 (2H,
+
+
m, C(O)NH), 7.71–7.77 (2H, m, C(O)NH). Mass spectrum: m/z 1375.7 [M + Na] , calcd 1375.8 [M + Na] . C H N O .
7
2 112 4 20
691

ACKNOWLEDGMENT
The work was sponsored by the Russian Science Foundation (Grant No. 14-50-00014).
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