Effective synthesis of novel C(2)-propargyl derivatives of betulinic and ursolic acids and their conjugation with β-d-glucopyranoside azides via click chemistry

Article abstract of DOI:10.1016/j.tet.2016.01.024

A new synthetic approach to the incorporation of an alkynyl group into pentacyclic triterpenoids was developed and utilized to prepare C(2)-propargyl derivatives of lupane- and ursane-type triterpenoic acids. Novel triterpenoid derivatives with a propynyl group at the C(2) atom were successfully used for click chemistry bioconjugation with glucopyranoside azides.

Full text of DOI:10.1016/j.tet.2016.01.024

Accepted Manuscript
Effective synthesis of novel С(2)-propargyl derivatives of betulinic and ursolic acids
and their conjugation with β-D-glucopyranoside azides via click chemistry
Anna Yu. Spivak, Rinat R. Gubaidullin, Zulfiya R. Galimshina, Darya A. Nedopekina,
Victor N. Odinokov
PII:
S0040-4020(16)30024-2
10.1016/j.tet.2016.01.024
TET 27431
DOI:
Reference:
To appear in: Tetrahedron
Received Date: 21 September 2015
Revised Date: 5 January 2016
Accepted Date: 14 January 2016
Please cite this article as: Spivak AY, Gubaidullin RR, Galimshina ZR, Nedopekina DA, Odinokov VN,
Effective synthesis of novel С(2)-propargyl derivatives of betulinic and ursolic acids and their conjugation
with β-D-glucopyranoside azides via click chemistry, Tetrahedron (2016), doi: 10.1016/j.tet.2016.01.024.
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ACCEPTED MANUSCRIPT
Effective synthesis of novel С(2)-propargyl derivatives of betulinic and ursolic acids and their
conjugation with β-D-glucopyranosideazides via click chemistry
Anna Yu. Spivak,* Rinat R. Gubaidullin, Zulfiya R. Galimshina, Darya A. Nedopekina, Victor N.
Odinokov

1
ACCEPTED MANUSCRIPT
Effective synthesis of novel С(2)-propargyl derivatives of betulinic and ursolic
acids and their conjugation with β-D-glucopyranoside azides via click chemistry
*
Anna Yu. Spivak, Rinat R. Gubaidullin, Zulfiya R. Galimshina,
Darya A. Nedopekina, Victor N. Odinokov
Institute of Petrochemistry and Catalysis, Russian Academy of Sciences, 141 prosp. Oktyabrya,
4
50075 Ufa, Russian Federation
Key words:
Pentacyclic triterpenoids, Alkylation of ketones, Metal enolates, Glucopyranosides,
Click chemistry.
Abstract:
A new synthetic approach to the incorporation of an alkynyl group into
pentacyclic triterpenoids was developed and utilized to prepare С(2)-propargyl
derivatives of lupane- and ursane-type triterpenoic acids. Novel triterpenoid
derivatives with a propynyl group at the С(2) atom were successfully used for click
chemistry bioconjugation with glucopyranoside azides.
1
. Introduction
Lupane- and ursane-type pentacyclic triterpenoids (betulin, betulinic and ursolic
acids) are abundant in Nature, being present in plants, and are of interest for medical
research for their diverse biological activities. These plant metabolites have been
found to exhibit antiviral, anti-inflammatory and, what is more important, antitumour
1
-4
properties, which are, moreover, combined with low animal toxicity. Owing to the
presence of easily transformable functional groups in these molecules, natural
pentacyclic triterpenoids have a high synthetic potential and are actively used as
*
Corresponding author. Tel./fax: +7 347 284 2750;
e-mail address: spivak.ink@gmail.com (Anna Yu. Spivak).

2
ACCEPTED MANUSCRIPT
promising structural platforms in synthetic transformations aimed at the development
of new drugs. However, the low solubility of natural triterpenoids in water and in
physiological fluids restricts their medical applications. In the last decade, numerous
semisynthetic betulinic and ursolic acid derivatives have been prepared by
modification of the functional groups at С(3), С(28), or С(30). In some cases, the
transformations gave compounds with higher bioavailability and higher biological
4
-9
activity than the parent compounds.
Transformations of ring А of triterpenoids at С(2) are promising but
considerably less studied. Betulinic acid analogue with a 1,3-enone moiety in ring А
having electron-withdrawing groups at С(2) (4) exhibited high antitumour activity and
1
0,11
inhibited nitrogen oxide production in activated macrophages (Fig.1).
Pronounced
cytotoxic activity towards various human cancer cells has been found for С(2)-
1
2
alkylidene branched lupane derivatives (5). Recently we prepared С(2)-conjugates of
betulinic and dihydrobetulinic acids with the triphenylphosphonium cation (6), which
1
3
were 40-50 times superior to betulinic acid in their antitumour activity. The C(2)-
triphenylphosphonium derivatives of lupane triterpenoids showed in vitro activity
against Schistosoma mansoni trematodes in low micromolar concentrations (IC =
5
0
1
4
0
.64-0.76 µg/ml).
Although the introduction of a triple bond into the betulinic and ursolic acid
molecule at С(3) or С(28) is known to enhance the synthetic and biological potentials
1
5-17
of these compounds,
no data on pentacyclic triterpenoids containing readily
functionalized alkynyl substituents at С(2) have been reported in the literature.

3
ACCEPTED MANUSCRIPT
3
0
30
2
9
20
2
9
20
3
19
21
1
9
21
22
12
1
2
17 22
13
1
17
18
1
8
5
25
11
26
2
5
11 26
CO Н
2
28
CO Н
CO
2
Н
14
16
2
14
16
28
15
1
1
2
9
10
1
9
8
2
3
10
8
A
4
27
3
A
4
5
7
6
27
5
7
6
HO
HO
O
23
3
1
24
23
2
24
-
Br
CO Me
CO Me
2
CO Н
2
+
2
NC
O
P
HO
O
6
5
4
Fig. 1. Chemical structures of betulinic acid (1), betulonic acid (2), ursolic acid (3) and lupane triterpenoids
4-6) with higher biological activities than the parent compounds.
(
2
. Results and Discussion
This communication reports the development of a chemoselective method for
the synthesis of new С(2)-propargyl lupane and ursane derivatives and the subsequent
transformation of these compounds into conjugates with 1,2,3-triazole
glucopyranosides.
The synthesis and pharmacological activities of natural and synthetic
triterpenoid О-glycosides (called saponins) have been the subjects of numerous
publications, as these secondary metabolites possess a broad spectrum of biological
18,19
and pharmacological activities.
Furthermore, binding of the hydrophilic sugar
moieties to terpenoid molecules may provide better water solubility, absorption, and
pharmacological properties. A large group of monodesmosidic and bisdesmosidic
triterpenoid saponins prepared using the different methods of glycosidic bond
1
8,20
formation at the С(3) or С(28) positions have been described to date.
The potential
of click reactions in the synthesis of triterpenoic acid–sugar biohybrids has not been
explored. Meanwhile, the click chemistry reactions of С(3)- or С(28)-alkynyl
derivatives of betulinic or ursolic acid with aromatic azides or azidopeptides were
successfully utilized to synthesize triazolyl-lupanes or triazolyl-ursanes as potential
17,21-23
anti-inflammatory and antitumour agents.

4
ACCEPTED MANUSCRIPT
The synthesis of the new alkynyl betulinic and ursolic acid derivatives 13 and 14
was performed via α-alkylation with propargyl bromide of the potassium
enoxytriethylborate resulting from elimination of the Н–С(2) proton of 3-ketolupane
or 3-ketoursane 7 and 8 by treatment with a strong base, KN(SiMe ) –Et B (Scheme
3
2
3
1
). The in situ generation of potassium enoxytriethylborate in the α-alkylation of
ketones serves to control the regio- and stereochemistry of these processes and reduces
the probability of formation of acetylene–allene rearranged and di- and polyalkylated
2
4,25
products.
We performed the reaction of propargyl bromide with enolate anion А, which
was formed by treating methyl betulonate 7 with KN(SiMe ) –Et B in 1,2-
3
2
3
dimethoxyethane at room temperature. Over a short period of time (1 h), the reaction
1
13
afforded the only diastereoisomer 9a ( H and C NMR data) with the equatorial-
oriented propynyl group. The С(2)-propynyl-substituted methyl ursonoate 10 was
synthesized in a similar way but using a DME–TНF (1:1) solvent mixture (Scheme 1).
The absolute configuration of the С(2)-carbon atom in compounds 9a and 10 was
determined using 2D NMR spectroscopy and the diagnostic Н–С(2) methine proton.
1
Indeed, the Н NOESY spectrum of compound 9a shows intense cross peaks of Н-С(2)
(
δH 2.84, HSQC, HMBC, and COSY spectra) with the axially oriented methyl groups
Ме(25) (δ 1.13) and Ме(24) (δ 1.05) and the proton Н–С(1) (δ 2.32, dd, J 13.5, 5.0
H
H
H
Hz). The observed stereochemistry of the С-alkylation of the enolate anion А, perhaps,
is attributable to steric reasons: the presence of the angular Ме(25) methyl group at the
С(10) atom increases the probability of the attack of the carbon electrophile on the
enolate ion from the sterically more accessible equatorial side.
O
O
25
CH3
H 2
4
H
H
CH3
1
2
O
H C
3
1
Fig. 2. H NOESY NMR experiments data of compound 9a.

5
ACCEPTED MANUSCRIPT
CO Н
CO Н
2
2
3
1
HO
HO
a,c 54%
63% b,c
CO Me
CO Me
2
2
8
7
O
O
d 88%
74% e
H
H
3'
2'
1'
CO Me
2
3
'
2'
1'
CO Me
2
2
5
1
1
0
9
a
O
O
A
4
10
B
6
4%
f
8
f
75%
-
5
+
K Et B O
3
24
23
A
CO Me
CO Me
2
2
11
12
HO
HO
7
4%
g
g 74%
CO H
2
CO H
2
1
3
HO
14
HO
o
o
Scheme 1. (a) CrO ,H SO , acetone, 0 C→20 °C, 5 h; (b) PCC, CH Cl , 0 C→20 °C, 8 h; (c) diazomethane,
3
2
4
2
2
o
Et O, 0 C→20 °C, 2 h; (d) KN(SiMe ) , Et B, propargyl bromide, DME, r.t., Ar, 1 h; (e) KN(SiMe ) , Et B,
2
3 2
3
3 2
o
3
.
o
propargyl bromide, DME–THF (1:1), r.t., Ar, 1 h; (f) NaBH , CeCl 7H O, MeOH—THF, –30 C→20 C, Ar,
4
3
2
2
h; (g) LiI, DMF, reflux, Ar, 2 h.
C(2)-Alkynyl betulinic and ursolic acids derivatives 13 and 14 were synthesized
from triterpenoids 9а and 10 in two-steps. The reduction of the keto group of
.
13
compounds 9а and 10 with CeCl 7H O-modified NaBH by a reported method
3
2
4
1
13
yielded 3β-epimers 11 and 12 with high stereoselectivity (95-96%, H and C NMR
3
data) (Scheme 1). The spin-spin coupling constant ( J
10.0 Hz) of the Н(3)
H(2),H(3)

6
ACCEPTED MANUSCRIPT
1
proton and the axial Н(2) proton found in the Н NMR spectra of compounds 11 and
2 attested to the axial position of Н(3) and, hence, the equatorial position (β-
orientation) of the ОН group. Demethylation of the sterically hindered ester group in
1
2
6
compounds 11 and 12 via halogenolysis with LiI in DMF as described previously
furnished С(2)-alkynyl betulinic and ursolic acid derivatives 13 and 14 in good yields
Scheme 1).
In relation to the alkylation of methyl betulonate 7 with propargyl bromide, it
(
was found that replacement of the DME solvent by TНF resulted in a considerably
lower yield of product 9a (11% in 1 h) due to incomplete conversion. When the
о
reaction temperature in DME was raised to 80 С, the reaction yielded a mixture of α-
and β-epimers 9а and 9b (Scheme 2), the β-epimer 9b content in the diastereoisomer
mixture increasing following the increase in the reaction time: 9а:9b=70:30 (1 h),
1
13
9
а:9b=50:50 (6 h), as shown by Н and С NMR data. The single epimer 9b was
isolated as a narrow chromatographic fraction from a mixture of diastereoisomers
difficult to separate, using column chromatography on SiO (hexane-ethyl acetate /
2
1
5
0:1). The Н NOESY spectrum of 9b showed no cross-peak for the methine Н(2)
proton with the H –Me(25) methyl protons but displayed intensive cross-peak with
ax
the protons of equatorial-oriented methyl group Me(23), which was indicative of the α-
orientation of H(2) and, hence, β-orientation of the propargyl moiety.
Elimination of the methine Н–С(2) proton in terpenoids 7 or 8 induced by
t
Bu OK in DME gave rise to potassium enolate B, which reacted with excess propargyl
t
bromide (7 or 8 : Bu OK : С Н Br molar ratio of 1:3:4.4) to give 2,2-bis-alkylated
3
3
products, terpenoids 15 and 16, in 53-58% yields (Scheme 2).

7
ACCEPTED MANUSCRIPT
30
2
9
20
3
1
9
21
22
1
2
a
1
1
7
1
8
5
1
1
26
+ 3'
CO Me
CO Me
3'
2'
1'
H
2'
1'
H
CO Me
2
1
4
16
2
2
50-55%
1
28
1
A
4
9
2
3
10
8
27
5
7
6
O
7
O
O
9а
9b
24
23
5
8%
b
9
9
а : 9b = 70 : 30 (1 h)
а : 9b = 50 : 50 (6 h)
25
1
CO Me
2
A
4
10
B
6
5
+
-
K O
1
5
O
23
24
B
c
3%
CO Me
CO Me
2
2
5
O
O
8
16
o
t
Scheme 2. (a) KN(SiMe ) , Et B, propargyl bromide, DME, 80 C, Ar; (b) Bu OK, propargyl bromide, DME,
3
2
3
t
r.t., Ar, 1 h; (c) Bu OK, propargyl bromide, DME–THF (1:1) r.t., Ar, 1 h
The absence of protons at the С(2) atom (δ 45.9 for compound 15 and δ 46.0 for
C
C
compound 16, DEPT and HSQC spectra) indicated that both alkynyl groups in
13
terpenoids 15 or 16 were attached to this atom. In the С NMR spectra, as might be
expected, the chemical shifts of geminal α- and β-oriented propargyl substituents
differed, being recorded, for example, for compound 15 at δ 28.6 and 31.0 (СН
C
2
group); δ 79.7 and 81.6 (С≡СН group); and δ 70.8 and 71.6 (С≡СН group).
C
C
The resulting С(2)-propargyl lupane and ursane triterpenoids derivatives 9-16
are «versatile» intermediates for the synthesis of new triterpenes. In this study, we
used these compounds for preparing the first representatives of betulinic and ursolic
acid conjugates with glucopyranosides linked at the С(2) atom through a 1,2,3-triazole
groups.

8
ACCEPTED MANUSCRIPT
3
CO Me
CO R
2
2
1
R
R
9b
O
2
1
2
3
9
1
1
a= R +R = C=O;
R = Me
1
2
3
1= R = OH; R = H; R = Me
1
2
3
3= R = OH; R = H; R = H
+
OR
O
N3
R= Ac, H
OR
RO
RO
click а 76-95%
OR
6'
3
OAc
4
'
CO R
6'
4
'
CO Me
2
O
5''
6''
2
5
'
4''
O
2'
5''
6''
2
'
5'
3'
4''
1
'
N
1'
RO
N
3'
AcO
RO
N N
OR
1
AcO
OAc
N N
R
2
O
R
23
1
2
3
1
7= R=Ac; R +R = C=O; R = Me
1
2
3
1
1
2
8= R=H; R +R = C=O; R = Me
1
2
3
9= R=Ac; R = OH; R = H; R = Me
1
2
3
0= R=H; R = OH; R = H; R = H
OR
O
N3
R= Ac, H
OR
OR
3
3
CO R
RO
O
CO R
2
2
RO
RO
N
N
RO
OR
1
N
1
R
R
R
2
click а 76-95%
2
R
1
2
3
21= R=Ac; R +R = C=O;
1
2
R = Me
3
1
1
0= R +R = C=O;
R = Me
1
2
3
22= R=H; R = OH; R = H;R = H
1
2
3
4= R = OH; R = H; R = H
OAc
O
OAc
N3
AcO
AcO
AcO
O
OAc
AcO
CO Me
2
N
N
CO Me
2
OAc
OAc
click а 86%
N
OAc
15
O
O
N
O
N
N
24
AcO
AcO
t
.
о
Scheme 3. (a) Bu OH, Cu powder, CuSO 5H O , 40 С, 17-23 h.
4
2
The β-D-glucopyranoside azides were obtained by the reaction of peracetylated
2
7
glucose with trimethylsilyl azide in the presence of FeCl , followed by deacetylation
3
28
t
with MeOH:H O:Et N (4:2:1). Coupling of alkynes and azides in Bu OH in the
2
3
.
4
о
presence of Cu powder and CuSO 5H O at 40 С for 17-23 h (TLC monitoring)
2
furnished bioconjugates 17-24 in 76-91% yields (Scheme 3). When sodium ascorbate

9
ACCEPTED MANUSCRIPT
10 mol %) and CuSO 5H O (5 mol %) were used as the catalysts and a Bu OH–Н О
.
t
(
4
2
2
served as the solvent, the yield of the target compounds did not exceed 30% due to the
low solubility of betulinic and ursolic acids in this medium. The formation of triazole
rings was validated by the characteristic resonance Н(5”) (δ 7.56-7.96), which
H
1
13
appeared in the Н NMR spectra of compounds 17-24. In the С NMR spectra, the
signals for С-(4’’) and С-(5’’) occurred at δ 143.6-147.6 and δ 120.3-123.7. The
C
C
resonance for the H(1’) anomeric protons in the glucoside moiety shifted downfield
with respect to the resonance of these protons in the starting azides (ꢀδ ~ 1.2); the
coupling constant for the anomeric protons (J ~ 7.6 Hz) is typical for a β-glycosidic
bond.
We have studied the steric effect of the propargyl substituents in epimers 9а and
9
b on their reactivity in the reaction with the acetylated glucopyranoside azide. In our
t
.
o
experiments (Bu OH, Cu, powder, CuSO 5H O, 40 C), epimer 9b was transformed
4
2
into conjugate 23 with ~ 91% conversion for over 14 h (the yield of compound 23 was
6% after purification by column chromatography on SiO ). The conversion of 9a
8
2
within this period of time was 12%. In this case, the reaction was completed only over
3 h.
2
The reaction between triterpenoid 15 and the acetylated glucopyranoside azide
occurred involving both the С(2)-α- and the С(2)-β-oriented propargyl substituents and
afforded the only bis-conjugate 24 in 86% or 43% yield, respectively, at the 1:2 or 1:1
1
terpenoid-glucoside ratio. The H NMR spectrum of compound 24 contained a double
set of signals corresponding to protons of triazole rings at the H-C(5''), protons H(1')-
H(6') and the methyl protons of the acetoxy groups, belonging to the sugar fragment.
13
In the С NMR spectrum a double set of signals manifested for carbon atoms С(4'')-
C(6''), carbon atoms of sugar moiety including the carbonyl and the methyl carbon
atoms of the acetoxy groups.

1
0
ACCEPTED MANUSCRIPT
3
. Conclusion
Chemoselective methods for the synthesis of С(2)-propargyl-substituted lupane-
and ursane-type triterpenoids were developed based on α-alkylation with propargyl
bromide of potassium enoxytriethylborates or potassium enolates generated from
t
betulonic or ursolic acid on treatment with KN(SiMe ) –Et B or Bu OK. The resulting
3
2
3
compounds are reactive building blocks for the synthesis of new potentially
biologically active terpenoids. С(2)-Propargyl-substituted pentacyclic triterpenoids
were successfully transformed to conjugates with 1,2,3-triazole-containing β-D-
glucopyranosides via Сu(I)-catalyzed 1,3-dipolar cycloaddition reaction (click
chemistry). Future study on Cu(I)-catalyzed 1,3-dipolar addition of pentacyclic
triterpenoids to other azide-sugars and evaluation of their antitumour activity of new
compounds are in progress.
4
4
. Experimental section
.1. General
IR spectra were recorded on a Specord IR–75 spectrometer (thin films or
1
13
solutions in CHCl ). H and C NMR spectra were recorded on a Bruker AVANCE–
3
1
13
5
00 instrument (500.13 ( H) and 125.78 MHz ( C)) in CDCl with Me Si as the
3
4
internal standard. Mass spectra of new compounds were recorded on a Bruker–
Autoflex III spectrometer (MALDI TOF, positive ion mode, 2,5-di-hydroxybenzoic
and α-cyano-4-hydroxycinnamic acids as the matrices) or on a LCMS-2010EV
(
Shimadzu) spectrometer of the UfIC RAS Center for Collective Use “Chemistry”.
Optical rotation was determined on a Perkin–Elmer-141 polarimeter. Specific rotation
α] is expressed in (deg mL)/(g dm); the concentration of the solution c is expressed
[
D
in g/100 mL. Elemental analysis was carried out on a Carlo Erba 1106 analyzer. TLC
was carried out on Sorbfil plates (Sorbpolimer, Krasnodar, Russia) in hexane–EtOAc
and chloroform–methanol (from 30:1 to 1:1); spots were visualized with anisaldehyde.
Silica gel L (KSKG grade, 50–160 ꢁm) was employed for column chromatography.
The starting compounds betulin, betulinic acid, ursolic acid and reagents: BEt (95%),
3

1
1
ACCEPTED MANUSCRIPT
KN(SiMe ) (1 M solution in THF), propargyl bromide, CeCl 7H O, NaBH , LiI,
.
3
3
2
2
4
CH COCl, TMSiN , Et N, sodium ascorbate, DMF, DME (dimethoxyethane),
3
3
3
(Aldrich). Alternatively betulonic and betulinic acids were obtained from betulin
29
according to the known procedures. Hexane, THF, and DME were heated under
reflux and distilled over sodium. The methyl esters of betulonic and ursolic acids were
3
0,31
prepared according to known procedures.
4
.2. Propargyl α-alkylation of compounds 7 and 8 in the system KN(SiMe ) – Et B
3
2
3
A 1 M solution of KN(SiMe ) (0.55 mL, 0.55 mmol) in THF was added under argon
3
2
at room temperature to a stirred solution of compound 7 or 8 (0.43 mmol) in DME or
DME–THF (1:1) (2 mL). After 15 min, 1 M Et B (0.60 mL, 0.60 mmol) in DME was
3
added and the mixture was stirred for 45 min. Then a solution of propargyl bromide
(0.07 mL, 0.85 mmol) was added. The reaction mixture was stirred for a specified
period of time (1-2 h, monitoring by TLC), neutralized with 3 M HCl (aq), and diluted
with water (1.5 mL). The product was extracted with EtOAc (4×10 mL). The
combined organic extracts were dried with MgSO and concentrated under reduced
4
pressure. The residue was purified by column chromatography on SiO with hexane–
2
EtOAc (from 30:1 to 1:1) as an eluent to give compound 9a or 10.
4
.2.1. Methyl-2α-propargyl-3-oxolup-20(29)-en-28-oate (9а). 0.19 g (88%), colorless
crystals, m.p. 57–59 °С; [Found: С, 80.6; H, 9.9. C H O requires С, 80.58; H,
34
50
3
1
9
–1
9
.94%]; [α]D –36.3 (c 0.9, CHCl ); ν (CHCl ) 3309 (≡C–H), 1724 (C=O) сm ; δ
3
max
3
H
(
500 MHz, CDCl ) 4.73, 4.59 (2 H, both br. s, H(29)), 3.68 (3 H, s, OMe), 3.01-2.98 (1
3
a
H, m, H(19)), 2.88-2.82 (1 H, m, H(2)), 2.60 (1 H, dt, J 16.5, 3.0 Hz, H (1'))), 2.32 (1
a
H, dd, J 13.5, 5.0 Hz, H (1)), 2.26-1.11 (21 H, m, CH, CH in pentacyclic skeleton and
2
b
1
H, H (1')), 1.95 (1 H, br.s, H(3')), 1.68 (3 H, s, H(30)), 1.13, 1.06, 1.05, 0.99, 0.95, (3
Н each, all s, Н(23)–Н(27)); δ (125 MHz, CDCl ) 215.7, 176.6, 150.4, 109.7, 82.9,
C
3
6
3
9.5, 57.3, 56.5, 51.3, 50.2, 49.4, 48.3, 46.9, 46.6, 42.5, 41.3, 40.8, 38.2, 37.4, 36.9,
+
4.1, 32.1, 30.5, 29.6, 25.4, 21.6, 21.2, 19.5, 19.3, 16.1, 14.6; MS: m/z [M + Na] ,
+
found 529.30. [C H O ] requires 506.38.
34
50
3

1
2
ACCEPTED MANUSCRIPT
β-propargyl-3-oxolup-20(29)-en-28-oate (9b). Colorless crystals, m.p.
4
.2.2. Methyl-2
2–84 °С; [Found: С, 80.6; H, 9.9. C H O requires С, 80.58; H, 9.94%]; [α] +101
1
D
9
8
3
4
50
3
–
1
(
c 0.7, CHCl ); ν (CHCl ) 3309 (≡C–H), 1724 (C=O) сm ; δ (500 MHz, CDCl )
3
max
3
H
3
4
.75, 4.62 (2 H, both br. s, H(29)), 3.67 (3 H, s, OMe), 3.03-2.99 (2 H, m, H(2),
a
H(19)), 2.54-2.51 (1 H, m, H (1')), 1.94 (1 H, br.s, H(3')), 1.70 (3 H, s, H(30)), 2.28-
b
1
0
8
3
.20 (22 H, m, CH, CH in pentacyclic skeleton and 1H, H (1')), 1.08, 1.04, 1.02, 0.92,
2
.68 (3 Н each, all s, Н(23)–Н(27)); δ (125 MHz, CDCl ) 218.4, 176.6, 150.5, 109.6,
C
3
2.8, 69.1, 56.6, 52.3, 51.3, 49.9, 49.4, 48.2, 46.9, 46.5, 42.4, 40.6, 40.4, 38.5, 36.9,
6.8, 32.9, 32.1, 30.6, 29.6, 29.2, 25.7, 22.0, 20.1, 19.9, 19.4, 18.5, 15.2, 15.3; MS: m/z
+
+
[
M + Na] , found 529.32. [C H O ] requires 506.38.
3
4
50
3
4
.2.3. Methyl-2α-propargyl-3-oxours-12-en-28-oate (10). 0.16 g (74%), colorless
crystals, m.p. 84–86 °С; [Found: С, 80.5; H, 9.8. C H O requires С, 80.58; H,
34
50
3
1
9
9
.94%]; [α]D +4 (c 0.6, CHCl ); ν (CHCl ) 3310 (≡C–H), 2119 (С≡С), 1722 (C=O)
3
max
3
–
1
сm ; δ (500 MHz, CDCl ) 5.27 (1 H, s, H(12)), 3.62 (3 H, s, OMe), 2.89-2.84 (1 H,
H
3
a
a
m, H(2)), 2.61 (1 H, dt, J 16.5, 3.0 Hz, H (1')), 2.32 (1 H, dd, J 13.0, 5.0 Hz, H (1)),
b
2
.30-1.13 (20 H, m, CH, CH in ursolic skeleton and 1 H, H (1')), 1.97 (1 H, br. s,
2
H(3')), 1.23, 1.09, 1.08, 1.07, 0.82 (3 Н each, all s, Н(23)–Н(27)), 0.95 (3 H, d, J 6.5
Hz, H(30)), 0.85 (3 H, d, J 6.5 Hz, H(29)); δ (125 MHz, CDCl ) 215.6, 177.9, 138.4,
C
3
1
3
1
4
25.1, 82.9, 69.5, 57.2, 52.8, 51.5, 48.1, 48.2, 47.1, 46.3, 42.1, 41.2, 39.6, 39.0, 38.9,
7.1, 36.6, 32.7, 30.6, 28.0, 25.2, 24.2, 23.5, 23.6, 21.9, 21.2, 19.5, 19.3, 17.1, 17.0,
+
+
5.6; MS: m/z [M + Na] , found 529.36. [C H O ] requires 506.38.
3
4
50
3
.
.3. Reduction of ketones 9a and 10 with NaBH –CeCl 7H O
4
3
2
.
A solution of CeCl 7H O (0.19 g, 0.5 mmol) in a mixture of THF–methanol (1:1) (2
3
2
mL) was added, via cannula for 3 min, a solution of compound 9a or 10 (0.4 mmol) in
a mixture of THF–methanol (1:2) (7.5 mL) cooled to –30 °C (Ar). Then, NaBH (0.03
4
g, 0.8 mmol) was added in small portions over 5 min, the temperature was raised to
~20 °C and the mixture was stirred at this temperature for 2 h (monitoring by TLC).
Then, the reaction mixture was neutralized with 5% aqueous HCl and extracted with
EtOAc (3×10 mL). The extract was washed sequentially with saturated aq. NaHCO3
and water, dried with MgSO , and the solvent was evaporated in vacuo. The residue
4

1
3
ACCEPTED MANUSCRIPT
was purified by column chromatography on SiO with hexane–EtOAc (from 30:1 to
2
1
:1) as an eluent to give compound 11 or 12.
4
.3.1. Methyl-2α-propargyl-3β-hydroxylup-20(29)-en-28-oate (11). 0.14 g (75%),
white crystals, m.p. 105–107 °С; [Found: С, 80.3; H, 10.2. C H O requires С, 80.26;
34
52
3
1
9
H, 10.30%]; [α]D –8.6 (c 1.1, CHCl ); ν (CHCl ): 3481 (–OH), 3308 (≡C–H), 1725
3
max
3
–
1
(
C=O) сm ; δ (500 MHz, CDCl ) 4.74, 4.60 (2 H, both br. s, H(29)), 3.68 (3 H, s,
H
3
OMe), 3.02 (1 H, d, J 10.0 Hz, H(3)), 3.00-2.88 (1 H, m, H(19)), 2.43 (1 H, dt, J 16.5,
3
0
1
.0 Hz, H(1')), 2.33 (1 H, ddd, J 16.5, 7.0, 3.0 Hz, H(1')), 1.99 (1 H, br. s, H(3')), 2.25-
a
.74 (22 H, m, CH, CH in pentacyclic skeleton), 1.84 (1 H, dd, J 15.0, 5.0 Hz, H (1)),
2
.69 (3 H, s, H(30)), 0.99, 0.98, 0.93, 0.87, 0.79 (3 Н each, all s, Н(23)–Н(27)) ; δC
(
125 MHz, CDCl ) 176.7, 150.5, 109.7, 83.0, 81.4, 69.9, 56.5, 55.4, 51.3, 50.5, 49.5,
3
4
6.9, 44.8, 42.4, 40.7, 39.1, 38.3, 37.3, 36.9, 34.8, 34.3, 32.2, 30.6, 29.6, 28.3, 25.5,
+
+
2
2.4, 20.9, 19.33, 18.5, 16.9, 16.2, 15.9, 14.7; MS (APCI): М , found 509. [C H O ]
34
52
3
requires 508.39.
4
.3.2. Methyl-2α-propargyl-3β-hydroxyurs-12-en-28-oate (12). 0.16 g (84%), white
crystals, m.p. 229–231 °С; [Found: С, 80.2; H, 10.2. C H O requires С, 80.26; H,
3
4
52
3
1
9
1
0.30%]; [α]D +31 (c 0.5, CHCl ); ν (CHCl ): 3545 (–OH), 3308 (≡C–H), 1715
3
max
3
–
1
(
C=O) сm ; δ (500 MHz, CDCl ) 5.27 (1 H, s, H(12)), 3.62 (3 H, s, OMe), 3.05 (1 H,
H
3
d, J 10.0 Hz, H(3)), 2.43 (1 H, dt, J 16.5, 3.0 Hz, H(1')), 2.36 (1 H, ddd, J 16.5, 5.0, 2.0
Hz, H(1')), 2.25 (1 H, d, J 10.0 Hz, H(18)), 2.02 (1 H, br. s, H(3')); 2.06-0.90 (21 H, m,
CH, CH in ursolic skeleton), 1.10, 1.02, 0.98, 0.82, 0.77 (3 Н each, all s, Н(23)–
2
Н(27)), 0.95 (3 H, d, J 7.0 Hz, H(30)), 0.89 (3 H, d, J 8.0 Hz, H(29)); δ (125 MHz,
C
CDCl ) 178.1, 138.2, 125.5, 83.0, 81.6, 69.9, 55.3, 52.9, 51.5, 48.1, 47.5, 44.7, 42.1,
3
3
1
4
9.5, 39.0, 38.9, 37.2, 36.6, 34.7, 32.9, 30.7, 28.5, 28.0, 24.2, 23.7, 23.4, 22.3, 21.2,
+
+
8.5, 17.0, 16.9, 16.5, 16.3; MS (APCI): М , found 509. [C H O ] requires 508.39.
34
52
3
.4. Halogenolysis of compounds 11 and 12 with LiI in DMF
LiI (0.70 g, 5 mmol) was added to a stirred solution of compound 11 or 12 (0.3 mmol)
in DMF (2 mL). The reaction mixture was heated under reflux for 2 h (monitoring by
TLC), diluted with water (2 mL), and neutralized with 5% HCl (aq). The product was
extracted with EtOAc (4×10 mL). The combined extracts were dried with MgSO and
4

1
4
ACCEPTED MANUSCRIPT
concentrate. The residue was purified by column chromatography on SiO with
2
hexane–EtOAc (from 30:1 to 1:1) as an eluent to give compound 13 or 14.
4
.4.1. 2α-Propargyl-3β-hydroxylup-20(29)-en-28-oic acid (13). 0.10 g (74%),
colorless crystals, m.p. 232–234 °С; [Found: С, 79.9; H, 10.2. C H O requires С,
3
3
50
3
1
9
8
0.11; H, 10.19%]; [α]D –17.3 (c 0.8, AcOEt); ν (thin film): 3306 (≡C–H), 1686
max
–
1
(
C=O) сm ; δ (500 MHz, CDCl ) 4.76, 4.62 (2 H, both br. s, H(29)), 3.05 (1 H, d, J
H
3
a
1
0.0 Hz, H(3)), 3.03-2.99 (1 H, m, H(19)), 1.85 (1 H, dd, J 13.0, 3.0 Hz, H (1)), 1.71
(
3 H, s, H(30)), 2.43-0.87 (22 H, m, CH, CH in pentacyclic skeleton and 1 Н, Н(3'), 2
2
Н, Н(1')), 1.00, 0.99, 0.96, 0.88, 0.79 (3 Н each, all s, Н(23)–Н(27)); δ (125 MHz,
C
CDCl ) 182.2, 150.4, 109.8, 82.9, 81.5, 69.9, 56.4, 55.4, 50.5, 49.3, 46.9, 44.8, 42.5,
3
4
1
0.7, 39.1, 38.4, 37.4, 37.1, 34.8, 34.2, 32.2, 30.5, 29.7, 28.3, 25.5, 22.4, 20.9, 19.3,
+
+
8.5, 16.9, 16.2, 16.0, 14.7; MS (APCI): М , found 493. [C H O ] requires 494.38.
33
50
3
4
.4.2. 2α-Propargyl-3β-hydroxyurs-12en-28-oic acid (14). 0.10 g (74%), colorless
crystals, m.p. 215–217 °С; [Found: С, 79.8; H, 10.2. C H O requires С, 80.11; H,
3
3
50
3
1
9
1
0.19%]; [α]D +16 (c 0.6, AcOEt); ν (thin film): 3552 (–OH), 3282 (≡C–H), 1690
max
–
1
(
C=O) сm ; δ (500 MHz, CDCl ) 5.25 (1 H, s, H(12)), 3.05 (1 H, d, J 10.0 Hz, H(3)),
H
3
2
.43 (1 H, dt, J 16.5, 2.5 Hz, H(1')), 2.36 (1 H, dd, J 16.5, 4.5 Hz, H(1')), 2.19 (1 H, d,
J 11.0 Hz, H(18)), 2.02 (1 H, br. s, H(3')), 2.10-0.91 (21 H, m, CH, CH in ursolic
2
skeleton), 0.96 (3 H, d, J 6.0 Hz, H(30)), 0.87 (3 H, d, J 6.5 Hz, H(29)), 1.10, 1.02,
0
1
3
.99, 0.80, 0.78 (3 Н each, all s, Н(23)–Н(27)); δ (125 MHz, CDCl ) 184.2, 137.9,
C
3
25.8, 83.0, 81.6, 70.0, 55.3, 52.5, 47.9, 47.4, 44.6, 41.9, 39.5, 39.0, 38.8, 37.2, 36.7,
4.6, 32.8, 30.6, 28.5, 27.9, 24.0, 23.7, 23.3, 22.3, 21.2, 18.5, 17.2, 16.9, 16.4, 16.3;
+
+
MS (APCI): М , found 493. [C H O ] requires 494.38.
3
3
50
3
4
.5. 2,2-Bis-propargylation of compounds 7 and 8
t
Bu OK (0.15 g, 1.29 mmol) was added at room temperature to a stirred solution of
compound 7 or 8 (0.43 mmol) in DME or DME–TНF (1:1) (2 mL). Then a solution of
propargyl bromide (0.16 mL, 1.89 mmol) in DME (1 mL) was added. The reaction
mixture was stirred for 1 h (monitoring by TLC), neutralized with 3 M HCl (aq), and
diluted with water (2 mL). The product was extracted with EtOAc (4×10 mL). The
combined extracts were dried with MgSO and concentrate. The residue was purified
4

1
5
ACCEPTED MANUSCRIPT
by column chromatography on SiO with hexane–EtOAc (from 30:1 to 1:1) as an
2
eluent to give compound 15 or 16.
4
.5.1. Methyl-2,2-bispropargyl-3-oxolup-20(29)-en-28-оаte (15). 0.14 g (58%),
colorless crystals, m.p. 78–81 °С; [Found: С, 81.6; H, 9.5. C H O requires С, 81.57;
3
7
52
3
2
0
–1
H, 9.62%]; [α]D +55.6 (c 0.8, CHCl ); ν (CHCl ): 3309 (≡C–H), 1724 (C=O) сm ;
3
max
3
δ (500 MHz, CDCl ) 4.75, 4.61 (2 H, both br. s, H(29)), 3.68 (3 H, s, OMe), 3.00-
H
3
a
a
2
2
.99 (1 H, m, H(19)), 2.86 (1 H, d, J 17.0 Hz, H (1')), 2.72 (1 H, d, J 17.0 Hz, H (1'')),
b
.58 (1 H, d, J 17.0 Hz, H (1'')), 2.05 (1 H, br. s, H(3')); 1.97 (1 H, br. s, H(3'')), 1.70
b
(
3 H, s, H(30)), 2.27-1.25 (22 H, m, CH, CH in pentacyclic skeleton and 1 Н, H (1')),
2
1
.19, 1.09, 1.01, 0.94, 0.75 (3 Н each, all s, Н(23)–Н(27)); δ (125 MHz, CDCl )
C
3
2
17.3, 176.6, 150.5, 109.6, 81.6, 79.7, 71.6, 70.8, 56.5, 51.6, 51.3, 50.9, 49.7, 49.4,
7.7, 46.9, 45.9, 42.5, 40.5, 38.4, 36.9, 36.8, 32.9, 32.1, 31.0, 30.6, 29.6, 28.6, 25.6,
4
+
+
2
3.1, 21.7, 20.3, 19.4, 17.2, 15.3, 14.7; MS: m/z [M + Na] , found 567.26. [C H O ]
37
52
3
requires 544.39.
4
.5.2. Methyl-2,2-bispropargyl-3-oxours-12-en-28-оаte (16). 0.12 g (53%), colorless
crystals, m.p. 85–88 °С; [Found: С, 81.5; H, 9.6. C H O requires С, 81.57; H,
37
52
3
1
9
–1
9
.62%]; [α]D +98 (c 0.5, CHCl ); ν (CHCl ): 3309 (≡C–H), 1722 (C=O) сm ; δ
3
max
3
H
(
500 MHz, CDCl ) 5.29 (1 H, s, H(12)), 3.60 (3 H, s, OMe), 2.86 (1 H, dd, J 17.0, 1.5
3
a
a
b
Hz, H (1')), 2.69 (1 H, dd, J 17.0, 1.5 Hz, H (1'')), 2.58 (1 H, d, J 17.0 Hz, H (1'')),
2
.03 (1 H, br. s, H(3')), 1.99 (1 H, br. s, H(3'')), 2.27-1.01 (21 H, m, CH, CH in ursolic
2
b
skeleton and 1 H, H (1')), 0.94 (3 H, d, J 6.0 Hz, H(30)), 0.86 (3 H, d, J 6.0 Hz,
H(29)), 1.20, 1.12, 1.10, 0.83, 0.77 (3 Н each, all s, Н(23)–Н(27)); δ (125 MHz,
C
CDCl ) 217.4, 177.9, 125.5, 138.1, 81.6, 79.7, 71.7, 70.9, 53.1, 51.9, 50.3, 48.2, 47.7,
3
4
2
6.3, 46.0, 42.3, 39.4, 39.1, 38.9, 36.6, 36.5, 31.9, 31.0, 30.7, 29.7, 28.7, 27.9, 24.2,
+
3.6, 23.5, 23.4, 21.2, 20.2, 17.0, 16.7, 16.6; MS: m/z [M + Na] , found 567.36.
+
[
C H O ] requires 544.39.
3
7
52
3
4
.6. General procedure for click reactions
The alkyne (0.20 mmol) and azide (0.25 mmol) were dissolved in t-BuOH (3 mL) then
.
4
Cu (7 mg, 0.11 mmol) and 1 M water solution of CuSO 5H O (0.04 mL, 0.04 mmol)
2
were added. The reaction mixture was stirred at 40 °C for 17-23 h (monitoring by

1
6
ACCEPTED MANUSCRIPT
TLC), then diluted with H O (4 mL) and extracted with EtOAc (4×10 mL). The
2
combined extracts were dried with MgSO and concentrate. The residue was purified
4
by column chromatography on SiO with hexane–EtOAc (from 3:1 to 1:1)
2
(
compounds 17, 19, 21, 23, 24) or CHCl –CH OH (from 4:1 to 1:1) (compounds 18,
3
3
2
4
0, 22) as an eluent to give compounds 17-24.
.6.1. Methyl 2α-[1N(2,3,4,6-tetraacetyl-β-D-glucopyranozo-1yl)-1H-1,2,3-triazole-4-
yl]-3-oxolup-20(29)-en-28-оаte (17). 0.19 g (95%), colorless crystals, m.p. 125–127
1
D
9
°
С; [Found: С, 65.4; H, 8.0. C H N O requires С, 65.51; H, 7.90%]; [α] –12 (c
48
69
3
12
–
1
1
.2, CHCl ); ν (CHCl ): 1757 (C=O) сm ; δ (500 MHz, CDCl ) 7.57 (1 H, s,
3
max
3
H
3
H(5'')), 5.81 (1 H, d, J 9.0 Hz, H(1')), 5.43-5.36 (2 H, m, H(2'), H(3')), 5.24 (1 H, t, J
0.0 Hz, H(4')), 4.71, 4.58 (2 H, both br. s, H(29)), 4.30 (1 H, dd, J 12.5, 5.0 Hz,
1
b
a
H (6')), 4.15 (1 H, dd, J 12.5, 1.5 Hz, H (6')), 3.99-3.96 (1 H, m, H(5')); 3.66 (3 H, s,
a
OMe), 3.15-3.07 (2 H, m, H (6''), H(2)), 2.98-2.95 (1 H, m, H(19)), 2.65 (1 H, dd, J
b
1
5.0, 6.0 Hz, H (6'')), 2.09 (3 H, s, COCH ), 2.04 (3 H, s, COCH ); 2.03 (3 H, s,
3
3
COCH ), 2.23-0.95 (22 H, m, CH, CH in pentacyclic skeleton), 1.84 (3 H, s, COCH ),
3
2
3
1
.66 (3 H, s, H(30)), 1.08, 1.06, 1.04, 0.95, 0.92 (3 Н each, all s, Н(23)–Н(27)); δC
(
125 MHz, CDCl ) 216.8, 176.6, 170.5, 169.9, 169.3, 168.6, 150.4, 146.9, 120.6,
3
1
09.7, 85.8, 75.1, 72.6, 70.3, 67.6, 61.5, 57.4, 56.4, 51.3, 50.1, 49.4, 48.4, 46.9, 46.8,
2.5. 41.9, 40.7, 38.2, 37.5, 36.9, 34.1, 32.1, 30.5, 29.6, 26.1, 25.4, 24.9, 21.6, 21.0,
4
+
+
2
0.5, 20.7, 19.3, 19.2, 16.1, 14.6; MS: m/z [M + Na] , found 902.39. [C H N O ]
4
8
69
3
12
requires 879.49.
4
.6.2.
Methyl
2α-[1N(2,3,4,6-tetrahydroxyl-β-D-glucopyranozo-1yl)-1H-1,2,3-
triazole-4-yl]-3-oxolup-20(29)-en-28-оаte (18). 0.12 g (85%), white crystals, m.p.
1
9
1
–
65–167 °С; [Found: С, 67.5; H, 8.6. C H N O requires С, 67.48; H, 8.64%]; [α]
4
0
61
3
8
D
–
1
14 (c 1.0, CHCl ); ν (CHCl ): 3391 (–OH); 1727 (C=O) сm ; δ (500 MHz,
3
max
3
H
CDCl ) 7.66 (1 H, s, H(5'')), 5.52 (1 H, d, J 7.5 Hz, H(1')), 4.73, 4.64 (2 H, both br. s,
3
H(29)), 4.08-4.03 (1 H, m, H(2')), 3.82-3.75 (4 H, m, H(3'), H(4'), H(6')), 3.69 (3 H, s,
a
OMe), 3.60-3.56 (1 H, m, H(5')), 3.15-3.12 (1 H, m, H(2)), 3.03-2.98 (2 H, m, H (6''),
b
H(19)), 2.49 (1 H, d, J 12.5 Hz, H (6'')), 2.24-1.04 (21 H, m, CH, CH in pentacyclic
2
a
skeleton), 2.11-2.09 (1 H, m, H (1)), 1.68 (3 H, s, H(30)), 0.99, 0.98, 0.97, 0.96, 0.95

1
7
ACCEPTED MANUSCRIPT
3 Н each, all s, Н(23)–Н(27)); δ (125 MHz, CDCl ) 217.7, 176.6, 150.3, 146.1,
(
C
3
1
4
1
7
4
22.9, 109.8, 87.7, 78.9, 76.8, 72.4, 68.8, 60.9, 57.5, 56.5, 51.3, 50.1, 49.4, 48.5, 48.1,
6.9, 42.5, 42.0, 40.7, 38.2, 36.9, 34.1, 32.2, 30.5, 29.6, 26.5, 25.4, 25.1, 21.4, 21.2,
+
+
9.3, 19.2, 16.2, 16.1, 14.7; MS: m/z [M + Na] , found 734.41. [C H N O ] requires
4
0
61
3
8
11.45.
.6.3. Methyl 2α-[1N(2,3,4,6-tetraacetyl-β-D-glucopyranozo-1yl)-1H-1,2,3-triazole-4-
yl]-3β-hydroxylup-20(29)-en-28-оаte (19). 0.16 g (91%), white crystals, m.p. 136–138
1
D
9
°
С; [Found: С, 65.4; H, 8.2. C H N O requires С, 65.36; H, 8.11%]; [α] –15 (c
48
71
3
12
–
1
1
.4, CHCl ); ν (CHCl ): 1757 (C=O) сm ; δ (500 MHz, CDCl ) 7.59 (1 H, s,
3
max
3
H
3
H(5'')), 5.85 (1 H, d, J 9.0 Hz, H(1')), 5.43-5.41 (2 H, m, H(3'), H(2')), 5.26-5.22 (1 H,
b
m, H(4')), 4.70, 4.57 (2 H, both br. s, H(29)), 4.31 (1 H, dd, J 12.5, 5.0 Hz, H (6')),
a
4
2
.11 (1 H, d, J 12.5 Hz, H (6')), 4.02-3.99 (1 H, m, H(5')), 3.66 (3 H, s, OMe), 2.98-
a
.96 (2 H, m, H (6''), H(19)), 2.76 (1 H, d, J 10.0 Hz, H(3)); 2.70 (1 H, dd, J 14.5, 7.0
b
Hz, H (6'')), 2.21-0.63 (23 H, m, CH, CH in pentacyclic skeleton), 2.06 (3 H, s,
2
COCH ), 2.05 (3 H, s, COCH ), 2.01 (3 H, s, COCH ), 1.83 (3 H, s, COCH ), 1.65 (3
3
3
3
3
H, s, H(30)), 0.95, 0.91, 0.87, 0.79, 0.77 (3 Н each, all s, Н(23)–Н(27)); δ (125 MHz,
C
CDCl ) 176.6, 170.4, 169.9, 169.4, 150.5, 147.0, 120.3, 109.6, 85.7, 81.6, 75.1, 72.6,
3
7
0.3, 67.8, 61.5, 56.5, 55.6, 51.2, 50.4, 49.5, 46.9, 45.0, 42.4, 40.7, 39.2, 38.2, 37.4,
6.9, 35.6, 34.3, 32.2, 30.5, 29.6, 28.9, 28.3, 25.5, 20.8, 20.7, 20.5, 20.4, 20.1, 19.3,
3
+
+
1
8.5, 16.8, 16.2, 15.9, 14.7; MS: m/z [M + Na] , found 904.43. [C H N O ]
48
71
3
12
requires 881.50.
4
3
.6.4. 2
α-[1N(2,3,4,6-tetrahydroxyl-β-D-glucopyranozo-1ил)-1H-1,2,3-triazole-4-yl]-
β-hydroxylup-20(29)-en-28-oic acid (20). 0.12 g (76%) white crystals, m.p. 210–212
1
D
9
°
С; [Found: С, 62.9; H, 8.7. C H N O requires С, 62.92; H, 8.78%]; [α] –23.3 (c
39
61
3
8
–
1
0
7
3
.9, MeOH); ν (thin film): 3344 (–ОН); 1693 (С=O) сm ; δ (500 MHz, MeOD)
max
H
.93 (1 H, s, H(5'')), 5.58 (1 H, d, J 10.0 Hz, H(1')), 4.71, 4.60 (2 H, both br. s, H(29)),
.95-3.89 (2 H, m, H(2'), H(6')), 3.75-3.72 (1 H, m, H(6')), 3.61-3.50 (3 H, m, H(3'),
H(4'), H(5')), 3.17 (1 H, d, J 11.5 Hz, H(6")), 3.06-3.00 (1 Н, m, H(19)), 2.83 (1 H, d, J
0.5 Hz, H(3)), 2.56-2.52 (1 H, m, H(6")), 2.33-0.62 (23 H, m, CH, CH in pentacyclic
1
2

1
8
ACCEPTED MANUSCRIPT
skeleton), 1.71 (3 H, s, H(30)), 1.01, 0.99, 0.96, 0.83, 0.82 (3 Н each, all s, Н(23)–
Н(27)); δ (125 MHz, MeOD) 180.4, 152.1, 147.6, 123.7, 110.3, 89.5, 83.0, 81.3, 78.7,
C
7
3
4.0, 71.0, 62.5, 57.6, 57.1, 52.1, 50.6, 48.6, 46.4, 43.7, 42.1, 40.5, 39.7, 38.5, 38.3,
7.4, 35.7, 33.5, 31.8, 30.9, 29.9, 29.2, 27.0, 22.2, 19.8, 19.7, 17.1, 17.6, 16.8, 15.3;
+
+
MS (APCI): [M] , found 700. [C H N O ] requires 699.45.
39
61
3
8
4
.6.5. Methyl 2
α-[1N(2,3,4,6-tetraacetyl-β-D-glucopyranozo-1yl)-1H-1,2,3-triazole-4-
yl]-3-oxours-12-en-28-оаte (21). 0.15 g (85%), white crystals, m.p. 134–136 °С;
1
D
9
[
Found: С, 65.5; H, 7.8. C H N O requires С, 65.51; H, 7.90%]; [α] +10 (c 0.5,
4
8
69
3
12
–
1
CHCl ); ν (CHCl ): 2948 (C=C); 1757 (C≡O); 1704 (СOOMe) сm ; δ (500 MHz,
3
max
3
H
CDCl ) 7.56 (1 H, s, H(5'')), 5.80 (1 H, d, J 9.0 Hz, H(1')), 5.41-5.32 (2 H, m, H(2'),
3
a
H(3')), 5.23-5.21 (2 H, m, H(12), H(4')), 4.28 (1 H, dd, J 12.5, 4.5 Hz, H (6')), 4.13 (1
b
H, d, J 12.5 Hz, H (6')), 3.99-3.97 (1 H, m, H(5')), 3.58 (3 H, s, OMe), 3.13-3.09 (2 H,
a
b
m, H (6''), H(2)), 2.67-2.63 (1 H, m, H (6'')), 2.18 (1 H, d, J 10.0 Hz, H(18)), 2.06 (3
H, s, COCH ), 2.04 (3 H, s, COCH ), 2.00 (3 H, s, COCH ), 1.98-0.70 (20 H, m, CH,
3
3
3
CH in ursolic skeleton), 1.81 (3 H, s, COCH ), 1.16, 1.06, 1.04, 1.01, 0.76 (3 Н each,
2
3
all s, Н(23)–Н(27)), 0.91 (3 H, d, J=5.0 Hz, H(30)), 0.81 (3 H, d, J=6.0 Hz, H(29)); δC
125 MHz, CDCl ) 216.7, 177.9, 170.5, 169.8, 169.3, 168.5, 146.9, 138.2, 125.2,
(
3
1
4
2
20.6, 85.8, 75.0, 72.5, 70.4, 67.8, 61.5, 57.2, 53.1, 51.4, 48.4, 48.0, 47.0, 46.3, 42.0,
1.9, 39.5, 28.9, 38.8, 37.2, 36.6, 32.7, 30.6, 28.0, 26.0, 25.1, 24.1, 23.6, 23.4, 21.8,
+
1.1, 20.7, 20.5, 20.1, 19.3, 17.1, 17.0, 15.5; MS: m/z [M + Na] , found 902.47.
+
[
C H N O ] requires 879.49.
4
8
69
3
12
4
3
.6.6. 2
α
-[1N(2,3,4,6-tetrahydroxyl-β-D-glucopyranozo-1ил)-1H-1,2,3-triazole-4-yl]-
β-hydroxyurs-12-en-28-oic acid (22). 0.11 g (79%), white crystals, m.p. 217–219 °С;
1
D
9
[
Found: С, 62.8; H, 8.7. C H N O requires С, 62.92; H, 8.78%]; [α] –8 (c 0.5,
3
9
61
3
8
–
1
MeOH); ν (thin film): 3353 (–ОН); 1695 (С=O) сm ; δ (500 MHz, MeOD) 7.96 (1
max
H
H, s, H(5'')), 5.60 (1 H, d, J 9.0 Hz, H(1')), 5.22 (1 H, s, H(12)), 3.94-3.89 (2 H, m,
H(2'), H(6')), 3.76-3.74 (1 H, m, H(6')), 3.61-3.53 (3 H, m, H(3'), H(4'), H(5')), 3.18 (1
H, d, J 12.5 Hz, H(6")), 2.86 (1 H, d, J 10.0 Hz, H(3)), 2.57-2.53 (1 H, m, H(6")),
2
.22-0.70 (22 H, m, CH, CH in ursolic skeleton), 1.12, 1.02, 0.99, 0.86, 0.85 (3 Н
2

1
9
ACCEPTED MANUSCRIPT
each, all s, Н(23)–Н(27)), 0.94 (3 H, s, H(30)), 0.89 (3 H, d, J 6.0 Hz, H(29)); δ (125
C
MHz, MeOD) 180.8, 146.1, 138.2, 125.4, 122.2, 88.0, 81.6, 79.7, 77.1, 72.6, 69.5,
6
2
1.0, 55.5, 52.9, 47.5, 47.3, 44.8, 41.9, 39.1, 39.0, 39.4, 38.9, 36.8, 35.7, 32.9, 30.5,
8.4, 27.9, 27.8, 24.0, 23.0, 22.9, 20.4, 18.4, 16.6, 16.4, 16.1, 15.5; MS: m/z [M +
+
+
Na] , found 722.47. [C H N O ] requires 699.45.
3
9
61
3
8
4
.6.7. Methyl 2
β-[1N(2,3,4,6-tetraacetyl-β-D-glucopyranozo-1yl)-1H-1,2,3-triazole-4-
yl]-3-oxolup-20(29)-en-28-оаte (23). 0.18 g (86%), colorless crystals, m.p. 138–140
1
D
9
°
С; [Found: С, 65.4; H, 7.8. C H N O requires С, 65.51; H, 7.90%]; [α] +47.5 (c
4
8
69
3
12
–
1
0
.8, CHCl ); ν (CHCl ): 1757 (C=O) сm ; δ (500 MHz, CDCl ) 7.57 (1 H, s,
3
max
3
H
3
H(5'')), 5.81 (1 H, d, J 9.0 Hz, H(1')), 5.42-5.38 (2 H, m, H(2'), H(3')), 5.24 (1 H, t, J
.5 Hz, H(4')), 4.72, 4.58 (2 H, both br. s, H(29)), 4.30 (1 H, dd, J 13.0, 5.0 Hz,
9
b
a
H (6')), 4.13 (1 H, dd, J 13.0, 1.5 Hz, H (6')), 4.00-3.97 (1 H, m, H(5')), 3.66 (3 H, s,
a
OMe), 3.23-3.17 (1 H, m, H(2)), 3.05 (1 Н, dd, J 15.0, 7.5 Hz, H (6'')), 3.10-2.96 (1 H,
b
m, H(19)), 2.60 (1 H, dd, J 15.0, 5.0 Hz, H (6'')), 2.09 (3 H, s, COCH ), 2.07 (3 H, s,
3
COCH ), 2.02 (3 H, s, COCH ), 2.24-0.92 (22 H, m, CH, CH in pentacyclic skeleton),
3
3
2
1
.83 (3 H, s, COCH ), 1.67 (3 H, s, H(30)), 1.03, 1.02, 0.99, 0.89, 0.67 (3 Н each, all s,
3
Н(23)–Н(27)); δ (125 MHz, CDCl ) 219.3, 176.6, 170.5, 169.9, 169.3, 168.5, 150.5,
C
3
1
4
2
47.0, 120.4, 109.6, 85.7, 75.1, 72.7, 70.3, 67.7, 61.5, 56.5, 52.2, 51.3, 49.9, 49.3,
8.7, 46.9, 46.7, 42.4, 41.5, 40.6, 38.5, 36.9, 32.9, 32.1, 30.6, 29.6, 28.8, 26.4, 25.6,
+
2.0, 20.7, 20.5, 20.2, 19.4, 18.6, 15.3, 15.1; MS (APCI): [M] , found 880.
+
[
C H N O ] requires 879.49.
4
8
69
3
12
4
.6.8.
Methyl-2,2-Bis-[1N(2,3,4,6-tetraacetyl-β-D-glucopyranozo-1yl)-1H-1,2,3-
triazole-4-yl]-3-oxolup-20(29)-en-28-оаte (24). 0.22 g (86%, реакцию проводили
при мольном соотношении алкин (15) : азид = 1:2), colorless crystals, m.p. 165–
1
9
1
1
67 °С; [Found: С, 60.5; H, 6.4. C H N O requires С, 60.45; H, 6.51%]; [α]
–
6
5
90
6
21
D
–
1
8.9 (c 0.6, CHCl ); ν (CHCl ): 1756 (C=O) сm ; δ (500 MHz, CDCl ) 7.90, 7.53
3
max
3
H
3
(2 H, s, H(5''), 5.83 (1 H, d, J 9.5 Hz, H(1')), 5.80 (1 H, d, J 9.5 Hz, H(1')), 5.51 (1 H, t,
J 9.5, H(2')), 5.49 (1 H, t, J 9.5, H(2')), 5.41 (2 H, t, J 9.5 Hz, H(3')), 5.28-5.25 (2 H,
b
m, H(4')), 4.71, 4.56 (2 H, both br. s, H(29)), 4.32-4.29 (2 H, m, H (6')), 4.19-4.17 (2
a
H, m, H (6')), 3.99-3.98 (2 H, m, H(5')), 3.66 (3 H, s, OMe), 3.11-0.93 (23 H, m, CH,

2
0
ACCEPTED MANUSCRIPT
CH in pentacyclic skeleton and 4 H, H(6'')), 2.10, 2.09 (3 H each, both s, COCH ),
2
3
2
.07 (6 H, s, COCH ), 2.03 (6 H, s, COCH ), 1.91 (3 H, s, COCH ), 1.83 (3 H, s,
3
3
3
COCH ), 1.65 (3 H, s, H(30)), 1.25, 1.06, 0.98, 0.89, 0.47 (3 Н each, all s, Н(23)–
3
Н(27)); δ (125 MHz, CDCl ) 219.5, 176.6, 170.54, 170.51, 169.98, 169.96, 169.34,
C
3
1
7
5
3
2
69.33, 168.97, 168.74, 150.3, 145.21, 143.55, 122.79, 121.30, 109.68, 85.71, 85.60,
5.10, 74.98, 72.81, 72.67, 70.52, 70.11, 67.81, 67.78, 61.60, 61.58, 56.45, 51.88,
1.28, 49.91, 49.47, 49.37, 46.86, 46.16, 42.47, 40.44, 38.38, 37.43, 36.93, 36.68,
3.01, 32.92, 32.08, 30.63, 30.49, 29.53, 25.65, 23.17, 21.68, 20.72, 20.68, 20.54,
+
0.51, 20.22, 20.14, 19.98, 19.30, 16.28, 15.28, 14.61; MS (APCI): [M] , found 1291.
+
[
C H N O ] requires 1290.62.
6
5
90
6
21
Acknowledgments
This work was financially supported by the Russian Foundation for Basic Research
(Project No. 16-33-60008).
References and notes
1
.
Tolstikov, G. A.; Flekhter, O. B.; Shul´ts, E. E.; Baltina, L. A.; Tolstikov, A. G.
Chemistry in the Interest of Stable Development. 2005, 13, 1.
2
3
.
.
Cichewicz, R. A.; Kouzi, S. A. Med. Res. Rev. 2004, 24, 90.
Mukherjee, R.; Kumar, V.; Srivastava, S. K.; Agarwal, S. K.; Burman, A. C.
Anti-Cancer Agents Med. Chem. 2006, 6, 271.
Sarek, J.; Kvasnica, M.; Vik, M.; Urban, M.; Dzubak, P.; Hajduch, M. In
4
.
Research on Melanoma - A Glimpse into Current Directions and Future Trends;
Murph, M., Ed.; In Tech, 2011; Vol. 7, pp 125-158.
5
2
6
.
Santos, R. C.; Salvador, J. A. R.; Marin, S.; Cascante, M. Bioorg. Med. Chem.
009, 17, 6241.
Suresh, C.; Zhao, H.; Gumbs, A.; Chetty, C.S.; Bose, H.S. Bioorg. Med. Chem.
.
Lett. 2012, 22, 1734.

2
1
ACCEPTED MANUSCRIPT
Kommera, H.; Kaluderovic, G. N.; Kalbitz, J.; Drager, B.; Paschke, R. Eur. J.
7
.
Med. Chem. 2010, 45, 3346.
8
9
.
.
Meng, Y.; Song, Y.; Yan, Z.; Xia, Y. Molecules. 2010, 15, 4033.
Liu, M.-C.; Yang, S.-J.; Jin, L.-H.; Hu, D.-Y.; Xue, W.; Song, B.-A.; Yang, S.
Eur. J. Med. Chem. 2012, 58, 128.
0. Honda, T.; Liby, K.T.; Su, X.; Sundararajan, C.; Honda, Y.; Suh, N.;
1
Risingsong, R.; Williams, C.R.; Royce, D.B.; Sporn M.B.; Gribble G.W. Bioorg. Med.
Chem. Lett. 2006, 15, 6306.
1
3
1
2
1
1. You, Y-J.; Kim, Y.; Nam, N-H.; Ahn B-Z. Bioorg. Med. Chem. Lett. 2003, 13,
137.
2. Csuk, R.; Stark, S.; Nitsche, C.; Barthel, A.; Siewert, B. Eur. J. Med. Chem.
012, 53, 337.
3. Spivak, A. Yu.; Nedopekina, D. A.; Shakurova, E. R.; Khalitova, R. R.;
Gubaidullin, R. R.; Odinokov, V. N.; Dzhemilev, U. M.; Bel´skii, Yu. P.; Bel´skaya,
N. V.; Stankevich, S. A.; Korotkaya, E. V.; Khazanov, V. A. Russ. Chem. Bull., Int.
Ed. 2013, 62, 188.
1
4. Spivak, A. Yu.; Keiser, J.; Vargas, M.; Gubaidullin, R. R.; Nedopekina, D. A.;
Shakurova, E. R.; Khalitova, R. R.; Odinokov, V. N. Bioorg. Med. Chem. 2014, 22,
6
1
297.
5. Csuk, R.; Barthel, A.; Sczepek, R.; Siewert, B.; Schwarz, S. Arch. Pharm.
Chem. Life Sci. 2011, 1, 37.
1
4
1
6. Csuk, R.; Sczepek, R.; Siewert, B.; Nitsche, C. Bioorg. Med. Chem. 2013, 21,
25.
7. Majeed, R.; Sangwan, P. L.; Chinthakindi, P. K.; Khan, I.; Dangroo, N. A.;
Thota, N.; Hamid, A.; Sharma, P. R.; Saxena, A. K.; Koul, S. Eur. J. Med. Chem.
2
1
2
1
013, 63, 782.
8. Gauthier, C.; Legault, J.; Piochon-Gauthier, M.; Pichette, A. Phytochem. Rev.
011, 10, 521.
9. Netala, V. R.; Ghosh, S. B.; Bobbu, P.; Anitha, D.; Tartte, V. Int. J. Pharm.
Pharm. Sci. 2015, 7, 24.

2
2
ACCEPTED MANUSCRIPT
0. Gauthier, C.; Legault, J.; Lebrun, M.; Dufour, P.; Pichette, A. Bioorg. Med.
2
Chem. 2006, 14, 6713.
1. Govdi, A. I.; Vasilevsky, S. F.; Nenajdenko, V. G.; Sokolova, N. V.; Tolstikov,
G. A. Russ. Chem. Bull., Int. Ed. 2011, 11, 2401.
2. Vasilevsky, S. F.; Govdi, A. I.; Sorokina, I. V.; Tolstikova, T. G.; Baev, D. S.;
Tolstikov, G. A.; Mamatuyk V. I.; Alabugin I. V. Bioorg. Med. Chem. Lett. 2011, 1,
2
2
6
2
2.
3. Govdi, A. I.; Sokolova, N. V.; Sorokina, I. V.; Baev, D. S.; Tolstikova, T. G.;
Mamatuyk, V. I.; Fadeev, D. S.; Vasilevsky, S. F.; Nenajdenko, V. G. Med. Chem.
Comm. 2015, 6, 230.
2
2
2
2
2
4. Negishi, E.; Idacavage, M. J.; Tetrahedron Lett. 1979, 10, 845.
5. Negishi, E.; Chatterjee, H. M. S.; John, R. A. J. Org. Chem. 1982, 47, 3190.
6. Dean, P. D. G. J. Chem. Soc. C. 1965, 6655.
7. Salunke, S. B.; Babu, N. S.; Chen, C.-T. Chem. Commun. 2011, 47, 10440.
8. Kommera, H.; Kaluderovic, G. N.; Bette, M.; Kalbitz, J.; Fuchs, P.; Fulda, S.;
Mier, W.; Paschke, R. Сhem.-Biol. Int. 2010, 185, 128.
9. Kim, D. S. H. L.; Chen, Z.; Nguyen, T.; Pezzuto, J. M.; Qiu, Sh.; Lu, Z-Z.
Synthetic. Commun. 1997, 9, 1607
2
3
6
3
0. Urban, M.; Sarek, J.; Klinot, J.; Kornikova, G.; Hajduch, M. J. Nat. Prod., 2004,
7, 1100.
1. Ma, C. M.; Cai, S. Q.; Cui, J. R.; Wang, R. Q.; Tu, P. F.; Hattori, M.;
Daneshtalab, M. Eur. J. Med. Chem., 2005, 40, 582.