Preparation and conformational study of B-ring substituted lupane derivatives

Article abstract of DOI:10.1135/cccc20061131

New lupane-type triterpenoids with 5(6) double bond were prepared using the method of partial demethylation on carbon C-4. Hydroboration of the double bond led to 6α-hydroxy derivative. By the oxidation and following reduction of 6α-hydroxy derivative the

Full text of DOI:10.1135/cccc20061131

B-Ring Substituted Lupane Derivatives
1131
PREPARATION AND CONFORMATIONAL STUDY OF
B-RING SUBSTITUTED LUPANE DERIVATIVES+
a
Martin DRAČÍNSKÝa1, Simona HYBELBAUEROVÁa2, † Jan SEJBAL and
b,
Miloš BUDĚŠÍNSKÝ *
a
Department of Organic Chemistry, Faculty of Science, Charles University,
1
2
128 40 Prague 2, Czech Republic; e-mail: martindraca@yahoo.com, hybsim@hotmail.com
Institute of Organic Chemistry and Biochemistry, Academy of Sciences of the Czech Republic,
166 10 Prague 6, Czech Republic; e-mail: milos.budesinsky@uochb.cas.cz
b
Received February 13, 2006
Accepted April 15, 2006
Dedicated to Professor Antonín Holý on the occasion of his 70th birthday.
New lupan e-type triterpen oids with 5(6) double bon d were prepared usin g th e m eth od of
partial dem eth ylation on carbon C-4. Hydroboration of th e double bon d led to 6α-h ydroxy
derivative. By th e oxidation an d followin g reduction of 6α-h ydroxy derivative th e 6-oxo an d
6β-h ydroxy derivatives were prepared. A n ew m eth od for selective oxidation of secon dary
h ydroxy group in th e presen ce of prim ary h ydroxy group was perform ed. Th e con form ation
of rin g A of n ew lupan e-type 3-oxo derivatives with a substituen t on rin g B was elucidated
on th e bases of 1H an d 13C NMR spectra an d m olecular m odellin g.
Keyw ord s: Triterpen oids; Triterpen es; Hydroboration ; Oxidation ; Lupan e; NMR spectros-
copy; Con form ation an alysis.
Triterpen es are com m on con stituen ts of h igh er plan ts2. Recen tly, in terest-
in g an ti-HIV an d an ti-can cer activities h ave been foun d for som e of th em
(e.g. refs3–5). Most of th ese com poun ds perspective for m edical use are de-
rived from th e lupan e skeleton . B-rin g substituted lupan e derivatives are
quite rare in th e n ature an d n o syn th etic route leadin g to th em h as been
publish ed yet. Isolation s of 41 n atural products with th e B-rin g substituted
lupan e skeleton h ave been publish ed so far6–36. Som e of th em sh ow in ter-
estin g biological activities, for exam ple an tibacterial10,35, h epatoprotective37
+ Part CXIV in th e series Triterpen es; for part CXIII see ref.1
Collect. Czech. Chem. Commun. 2006, Vol. 71, No. 8, pp. 1131–1160
© 2006 Institute of Organic Chemistry and Biochemistry
doi:10.1135/cccc20061131

1132
Dračínský, Hybelbauerová, Sejbal, Buděšínský:
an d cytostatic13,30. An appropriate startin g com poun d with lupan e skeleton
is betulin , wh ich is easy to isolate from birch bark.
A m eth od of partial dem eth ylation at carbon C-4 followed by m eth yl-
ation of th is carbon with sim ultan eous in troduction of double bon d to
position 5 was ch osen . For th e preparation of B-rin g substituted lupan e de-
rivatives, dih ydrobetulin (1) was con verted in ten reaction steps to un satu-
rated n orketon e 14. 3-Oxolupan e-28-yl acetate (2) was th e key in term ediate
of th is reaction route (Sch em e 1). Th ere are two possible routes for prepara-
CH2OR2
(iii)
CH2OR
CN
R1
R1
R2
1, β-OH OH
4, R = Ac
8, R = Bz
(iv)
(i)
2, =O
3, =O
OAc
(ii)
OH
(v)
CH2OR3
(vi)
CH2OR
CN
O
O
R1
R1
6, CH3
10, CH3
11, H
R2
H
R3
R2
Ac
Bz
5, R = Ac
9, R = Bz
H
CH3 Bz
12, CH3 CHO Bz
(vi)
(vii)
CH2OR
O
O
7
13, R = Bz
14, R = H
(viii)
17, R = CH3
(i) Br2, Py, –24 °C; (ii) Ac2O, Py; (iii) see ref.39; (iv) 1. NaOH, EtOH 2. benzoyl chloride, Py;
(v) 3-chloroperoxybenzoic acid, CHCl3, 0 °C; (vi) 1. BF3Et2O, 2. 2 M HCl; (vii) HBr, AcOH, Br2;
(viii) KOH, EtOH
SCHEME 1
Collect. Czech. Chem. Commun. 2006, Vol. 71, No. 8, pp. 1131–1160

B-Ring Substituted Lupane Derivatives
1133
tion of th is acetoxyketon e from dih ydrobetulin (1). Th e first con sists in se-
lective acetylation of 28-h ydroxy group an d subsequen t oxidation of th e
3β-h ydroxy group. Th is partial acetylation is kn own 38 but th e yields are
quite poor an d th e product m ust be purified by colum n ch rom atograph y.
We decided to use th e oth er path way: selective oxidation of th e secon dary
3β-h ydroxy group an d subsequen t acetylation of th e h ydroxy group in po-
sition 28. As an oxidation agen t we ch ose aqueous solution of sodium
h ypoch lorite in acetic acid. Th e selectivity of th e oxidation reaction at
room tem perature was good but th e low solubility of diol 1 required h uge
am oun ts of acetic acid. Wh en th e reaction was carried out at h igh er tem -
peratures, th e solubility of th e startin g com poun d was h igh er but th e selec-
tivity was lower. An oth er oxidation agen t was brom in e in pyridin e. Best
results were ach ieved wh en th e reaction was carried at –24 °C. Th e selectiv-
ity of th is reaction was h igh an d th e preparative yield of 28-h ydroxylupan -
3-on e (3) was h igh er th an 90%. Th e h ydroxyketon e 3 was acetylated to
acetoxyketon e 2. Th e con version of th e com poun d 2 to secon itrile 4 was
perform ed accordin g to ref.39 Th e n ext step, epoxidation of secon itrile 4,
provided a m ixture of 4R an d 4S epoxy derivatives 5. Th e furth er reaction
was closure of rin g A an d decarbon ylation , with n orketon e 6 as th e ex-
pected product. Th is reaction is kn own in steroid ch em istry40 an d it was
successfully used also for preparation of olean an e derivatives41. We applied
th is reaction to th e epoxy derivatives 5 but th e yields of 3-oxo-24-n or-
lupan -28-yl acetate (6) were quite low (15%) sin ce th e solvolysis of acetoxy
group in position 28 an d furth er rearran gem en ts of rin g E led to product 7.
Its structure was con firm ed by IR, MS an d 2D-NMR spectroscopy. Wh en
oth er Lewis acid (AlCl3, FeCl3, Zn Cl2) was used, n o traces of n orketon e 6
were foun d in th e reaction m ixture. We decided to use an oth er protective
group of th e h ydroxy group in th e position 28. Acetoxysecon itrile 4 gave
ben zoylsecon itrile 8 after alkalin e h ydrolysis an d reaction with ben zoyl
ch loride. Secon itrile 8 reacts with 3-ch loroperoxyben zoic acid to give a
m ixture of epoxides 9. Wh en a m ixture of epoxyn itriles 9 was refluxed in
toluen e with boron trifluoride eth erate on ly sm all traces of products with
rearran ged rin g E appeared. Th e reaction m ixture con tain ed two m ain prod-
ucts. Th e first product was 3-oxo-24-n orlupan -28-yl ben zoate (10), wh ose
structure was con firm ed by m ass, in frared an d NMR spectroscopy (in
1H NMR spectrum a doublet of 4α-m eth yl group at δ 0.98). Th e sign als of
all h ydrogen s an d carbon s of com poun d 10 were assign ed usin g two-
dim en sion al NMR tech n iques (Tables I an d IV). Attem pts to isolate th e sec-
on d product failed because it is tran sform ed to n orketon e 10 durin g ch ro-
1
m atograph y. H NMR data of th is product were obtain ed from th e spectrum
Collect. Czech. Chem. Commun. 2006, Vol. 71, No. 8, pp. 1131–1160

1134
Dračínský, Hybelbauerová, Sejbal, Buděšínský:
of th e reaction m ixture, suggested structure of th e by-product is 4β isom er
(11) of n orketon e 10 (doublet of 4β-m eth yl group at δ 1.11). Steric in terac-
tion of th e 4β-m eth yl group with th e 10β-m eth yl group can decrease th e
th erm odyn am ic stability of com poun d 11 an d explain s its easy isom er-
isation to n orketon e 10. A sm all am oun t of 3,24-dioxolupan -28-yl ben zoate
(12) was also isolated from th e reaction m ixture. Its structure was con -
firm ed by m ass, in frared an d NMR spectroscopy (see Tables I an d IV). In
1H NMR spectrum a sign al of aldeh ydic h ydrogen (δ 9.71) appears; sign als
of all h ydrogen s an d carbon s of com poun d 12 were assign ed usin g 2D-NMR
tech n iques an d th e 4β position of th e aldeh ydic group was con firm ed by
NOESY spectrum . Norketon e 10 was brom in ated an d th en deh ydro-
brom in ated (with out isolation of th e brom o derivative) to give un saturated
n orketon e 13. Th e ben zoyl ester 13 was h ydrolysed to th e un saturated
keton e 14.
It is kn own 42 th at m eth ylation of 3-oxosteroid derivatives with m eth yl
iodide after en olisation with potassium tert-butyl alcoh olate takes place
preferen tially at th e C-4 an d m ixtures of 4-m on om eth yl an d 4,4-dim eth yl
derivatives are usually obtain ed. Th is reaction was also applied to an
olean an e derivative1. It was sh own th at n eith er prolon gation of th e reac-
tion with potassium tert-butoxide n or prolon gation of th e m eth ylation
in creased th e yields of th e 4,4-dim eth yl derivative. Wh en h igh er con cen tra-
tion s of potassium tert-butoxide an d m eth yl iodide were used, by products
with on e or two m eth yl groups in position 2 appeared1. Meth ylation of
n orketon e 14 with m eth yl iodide (Sch em e 2) after refluxin g with potassium
tert-butyl alcoh olate gave a m ixture of four com poun ds: startin g keton e 14
(38%), 4,4-dim eth yl keton e with double bon d in position 5 (15, 34%) an d
sm all am oun ts of two an alogous com poun ds with m eth oxy group in posi-
tion 28 (16, 5% an d 17, 5%). Keton e 15 was reduced with sodium
boroh ydride an d th e resultin g dih ydroxy derivative 18 was acetylated to
diacetoxy derivative 19. In IR spectra of 3-oxo derivatives 15 an d 16 on e
can fin d a weak absorption ban d of C=C valen ce vibration at 1666 cm –1
,
h ydroxy derivative 18 an d acetoxy derivative 19 h ave th is ban d very weak.
Double bon d in position 5 gives in 13C NMR spectra of all derivatives two
sign als – sin glet at δ 145–147 (C-5) an d doublet at δ 119–121 (C-6). In
1H NMR spectra H-6 appears as doublet of doublets at δ 5.50 with couplin g
con stan ts J(6,7β) = 5–6 an d J(6,7α) = 2–3 Hz, sign al of th e axial H-7α is easy
to iden tify at δ 2.20 as a very broad doublet with large gem in al couplin g
con stan t (17–18 Hz). Full assign m en t of sign als of all h ydrogen s an d car-
bon s of com poun ds 15, 16 an d 18 was m ade usin g 2D-NMR tech n iques
(Tables I, II an d V). In th e m ass spectra of keton es 15 an d 16 an ion at m/z
Collect. Czech. Chem. Commun. 2006, Vol. 71, No. 8, pp. 1131–1160

B-Ring Substituted Lupane Derivatives
1135
CH2OCH3
CH2OH
(i)
O
O
17
14
CH2OCH3
CH2OH
O
O
16
15
(ii)
(iv)
CH2OR
CH2OR
RO
RO
20, R = H
21, R = Ac
18, R = H
(iii)
OR
19, R = Ac
(v)
(vi)
CH2OH
CH2OPiv
R1O
23, R1 = R2 = H
24, R1 = Piv; R2 = H
25, R1 = H; R2 = Piv
O
22
OR2
OH
(vii)
(viii)
(vi)
CH2OR2
CH2OR
R1O
O
27, R1 = R2 = H
28, R1 = H; R2 = Ac
29, R1 = R2 = Ac
26, R = Piv
30, R = H
O
OH
(i) 1. t-BuOK, ∆, Ar, 2. MeI; (ii) NaBH4, C6H6, MeOH; (iii) Ac2O, Py; (iv) 1. BH3, THF, ∆p, 2. NaOH, H2O;
(v) pivaloyl chloride, Py, –10 oC; (vi) Br2, Py, –24 oC; (vii) CrO3, Py; (viii) 1. LiAlH4, Et2O, 2. AcOEt
SCHEME 2
Collect. Czech. Chem. Commun. 2006, Vol. 71, No. 8, pp. 1131–1160

1136
Dračínský, Hybelbauerová, Sejbal, Buděšínský:
124 appears, th is ion was foun d also in som e 4,4-dim eth yl steroid43 an d
triterpen oid derivatives1,7 with double bon d at position 5.
Th e double bon d in th e position 5 can be used for in troduction of vari-
ous substituen ts in to th e rin g B. In previously publish ed papers th e allylic
oxidation 44, allylic brom in ation 1 an d epoxidation 1,44 of triterpen oids with
double bon d in position 5 is described. In th is work we studied th e
h ydroboration of olefin 18, th at was prepared by reduction of keton e 15.
Th e h ydroboration did n ot work un der n orm al con dition s. Wh en per-
form ed at 80 °C in th e pressure vial th en after 8 h workin g up with h ydro-
gen peroxide in alkalin e m edium provided lupan e-3β,6α,28-triol (20) as th e
on ly product. Th e structure of th is com poun d follows from full assign m en t
1
of H an d 13C NMR ch em ical sh ifts usin g HSQC an d HMBC tech n iques (see
Tables II an d V). Th e stereoch em istry was con firm ed by an alysis of J cou-
plin gs an d NOESY spectrum . In 1H NMR spectrum proton H(6) appears as
a triplet of doublets at δ 4.05 with two large (alm ost iden tical) vicin al cou-
3
plin g con stan ts (10.6 Hz) an d on e sm aller (4.0 Hz). Th e J ~ 10.6 Hz in di-
cates trans-diaxial couplin g of H-6, wh ich m ust th erefore adopt position
6β an d th e h ydroxy group is in position 6α. In addition , th e h ydrogen
H-6 sh ows NOE con tacts (accordin g to NOESY spectrum , see Fig. 1) to th e
m eth yl groups 24, 25 an d 26, th at is possible on ly in th e case H-6 in th e po-
sition 6β.
Triol 20 con tain in g two secon dary an d on e prim ary h ydroxy group was
acetylated to triacetate 21. It was in terestin g to exam in e th e reaction of th e
triol with brom in e in pyridin e at –24 °C. Durin g th is reaction on ly th e 3β
h ydroxy group is oxidised wh ile th e 6β an d 28 h ydroxy groups do n ot re-
FIG. 1
Part of NOESY spectrum of com poun d 20. Th e NOE cross-peaks of H-6 (at δ 4.05) with m eth yl
proton s H-24, H-25 an d H-26
Collect. Czech. Chem. Commun. 2006, Vol. 71, No. 8, pp. 1131–1160

B-Ring Substituted Lupane Derivatives
1137
act; dih ydroxyketon e 22 is th e on ly reaction product. A possible reason for
distin ct reactivity of th e 3β an d 6β secon dary h ydroxy group is th e steric
h in dran ce of th e 6β h ydroxy group. To prepare 6-oxo derivatives we h ad to
use a stron ger oxidative agen t th an brom in e in pyridin e. First, th e 28 h y-
droxy group was protected by tran sform ation to pivaloyloxy derivative.
Wh en pivaloyl ch loride in pyridin e at –10 °C reacted with th e triol 20, pro-
tected derivative 23 appeared as a m ain product but sm all am oun ts of
dipivalates 24 an d 25 were also isolated from th e reaction m ixture.
Dih ydroxy derivative 23 was oxidised with ch rom ium trioxide to diketo de-
rivative 26. Th e pivalate protective group was rem oved by reduction with
lith ium alum in um h ydride. Durin g th is reaction , th e 3 an d 6 oxo groups
were also reduced an d lupan e-3β,6β,28-triol (27) was th e m ain product. To
decom pose th e residual lith ium alum in um h ydride eth yl acetate was used
an d som e tran sesterification took place an d 3β,6β-dih ydroxylupan -28-yl
acetate (28) was also isolated from th e reaction m ixture. Th e fact th at th e
ester of acetic acid appears on ly in th e position 28 is in agreem en t with th e
previous observation 38 th at prim ary h ydroxy group in position 28 of
lupan e derivatives is m ore reactive in esterification s th an secon dary h y-
droxy groups in oth er position s. Th e structures of com poun ds 27 an d 28
were con firm ed by m ass, IR an d NMR spectra. Both com poun ds sh ow in
1H NMR spectra a broad sign al at δ 4.53 (H-6). Sign als of all h ydrogen an d
carbon atom s of com poun d 28 were assign ed usin g 2D-NMR tech n iques
(Tables III an d VI). Triol 27 was acetylated at room tem perature to
6β-h ydroxylupan e-3β,28-diyl diacetate (29), 6β h ydroxy group did n ot react
un der th ese con dition s. We also studied selective oxidation of triol 27 with
brom in e in pyridin e at –24 °C. Un der th ese con dition s both th e 3β an d 6β
h ydroxy groups are oxidised, th e prim ary h ydroxy group in position 28
does n ot react an d dioxo derivative 30 is th e product. Sign als of all h ydro-
gen an d carbon atom s of com poun d 30 were assign ed usin g 2D-NMR tech -
n iques (see Tables III an d VI).
Olefin 18 was epoxidised with 3-ch loroperoxyben zoic acid to give
5α,6α-epoxylupan -3β,28-diol (31) (Sch em e 3). Th is epoxy derivative was
oxidised usin g brom in e in pyridin e at – 24 °C to epoxyketon e 32. Th e struc-
ture of epoxyketon e 32 follows from full assign m en t of all 1H an d 13C
sign als usin g HSQC, HMBC an d NOESY spectra. Th e 5α,6α con figuration of
th e epoxide is in agreem en t with th e previously observed epoxidation of
triterpen oid derivatives1,44. We also exam in ed th e reaction of th e diacetoxy
derivative 19 with N-brom oacetam ide. Th is reaction is used in steroid
ch em istry to prepare 5α-brom o-6β-h ydroxy derivatives45,46. In our h an ds,
olefin 19 gave after th e reaction with N-brom oacetam ide 5β,6β-epoxide 33
Collect. Czech. Chem. Commun. 2006, Vol. 71, No. 8, pp. 1131–1160

1138
Dračínský, Hybelbauerová, Sejbal, Buděšínský:
1
as th e on ly product. Its structure was con firm ed by full assign m en t of H
an d 13C NMR sign als by HSQC, HMBC an d NOESY spectra (Tables III an d
VI). Th e differen t reactivity of olefin 19 can be explain ed by th e steric h in -
dran ce of th e β side of th e B rin g of th e lupan e skeleton . Th e 6β-h ydroxy
group (wh ich is probably presen t in th e in term ediate) is in trans-position to
th e 5α-brom in e, th us in tram olecular epoxidation can take place an d th e re-
sultin g 5β,6β-epoxide is less sterically dem an din g th an th e 6β-h ydroxy de-
rivative in term ediate.
(i)
CH2OH
CH2OH
O
HO
HO
18, R = H
19, R = Ac
31
(iii)
(ii)
CH2OAc
CH2OH
O
O
AcO
O
33
32
(i) 3-chloroperoxybenzoic acid, CHCl3; (ii) 70% HClO4, N-bromoacetamide, CH2Cl2, EtOH, H2O;
(iii) Br2, Py, -24 °C
SCHEME 3
Conformation of Ring A
It is well kn own (e.g. ref.47) th at rin g A of 3-oxo derivatives of triterpen oids
can adopt oth er th an ch air con form ation s (twist-boat, sofa) depen din g on
oth er substituen ts. It was sh own 1,47 th at on e can determ in e th e con form a-
tion of th e rin g A on th e basis of an an alysis of vicin al couplin g con stan ts
of proton s in position s 1 an d 2. Th ree basic con form ation s of rin g A were
proposed in ref.48 Th ey are th e ch air con form ation C an d two twist-boat
con form ation s T1 an d T2. Later a sofa con form ation was foun d in crystal
structure of a 3-oxolupan e-28-n itrile49 (Fig. 2). Th e values of ch aracteris-
tic couplin g con stan ts of con form ation s C, T1, T2 an d S are presen ted in
Table VII togeth er with 3J(1,2) of 3-oxo derivatives prepared in th is work
Collect. Czech. Chem. Commun. 2006, Vol. 71, No. 8, pp. 1131–1160

B-Ring Substituted Lupane Derivatives
1139
an d allobetulon e47 (34). Th e values for C, T1 an d T2 are taken from ref.1, th e
values for S are calculated accordin g to a m odified Karplus equation 50. To
fin d out th e vicin al con stan ts it was n ecessary first to assign ch em ical sh ifts
O
O
34
of h ydrogen s in position s 1α, 1β, 2α an d 2β. For com poun ds 12, 15, 22, 30
an d 32 it was don e usin g NOESY spectra, for com poun d 26 accordin g to
an alogy with com poun d 30 an d for com poun d 16 accordin g to an alogy
with com poun d 15. It is discussed in ref.47 th at rin g A of 3-oxo derivatives
FIG. 2
Possible con form ation s of rin g
C(10)–C(1)–C(2)–C(3)
A of 3-oxo triterpen oids. Θ is th e torsion an gle
Collect. Czech. Chem. Commun. 2006, Vol. 71, No. 8, pp. 1131–1160

1140
Dračínský, Hybelbauerová, Sejbal, Buděšínský:
with out oth er substituen ts (e.g. allobetulon ) is foun d in an equilibrium of
ch air an d T1 con form ation .
Th e con form ation of rin g A of com poun ds 12, 15, 22 an d 30 was also
studied usin g m olecular m odellin g. It was don e with m olecules an alogical
to th ese com poun ds, wh ere isopropyl group in position 19 an d h y-
droxym eth yl or ben zoyloxym eth yl group in position 17 were replaced with
m eth yl groups. Th ese structural ch an ges sim plify th e calculation s an d are
sufficien tly far from rin g A to in fluen ce its con form ation . On e h un dred of
sim ulated an n ealin gs to 1000 K followed by slow coolin g to 200 K were per-
form ed with every m olecule. Th e calculated structures were th en sorted ac-
cordin g to th e con form ation of rin g A to several structural types. Th e
results of m olecular m odellin g are presen ted in Table VIII.
Couplin g con stan ts of aldeh yde 12 are very close to th ose of ch air con -
form ation an d th e m olecular m odellin g foun d also ch air con form ation as
th e global en ergy m in im a geom etry. Th e replacem en t of th e m eth yl group
in position 4β by th e carbaldeh yde group th us in creases th e stability of
ch air con form ation (in com parison with allobetulon e (34), wh ere equilib-
rium of C an d T1 con form ation takes place).
By exam in ation of th e couplin g con stan ts of derivatives with double
bon d in th e position 5 we can see th at th ey are in th e best agreem en t with
th e 3J(1,2) values of T2 con form er. Th e sam e con clusion was m ade for
olean an e derivatives in ref.1 Th e results of m olecular m odellin g are in good
agreem en t with th is con clusion . Figure 3 sh ows th e rin g A of th e lowest en -
ergy con form ation of m olecule an alogous to com poun d 15. Th e en ergy of
oth er con form ers is at least 26 kJ m ol–1 h igh er, th is m ean s th ey are alm ost
n ot presen ted in th e equilibrium .
FIG. 3
Rin gs A an d B of lup-5-en e derivatives (results of m olecular m odellin g)
Collect. Czech. Chem. Commun. 2006, Vol. 71, No. 8, pp. 1131–1160

B-Ring Substituted Lupane Derivatives
1141
Couplin g con stan ts of com poun d 22 with h ydroxy group in position 6α
agree best with th e values of T1 con form er. Th e sam e result was also ob-
tain ed by m olecular m odellin g. Figure 4 sh ows rin g A of th e lowest en ergy
con form ation of a m olecule an alogous to 6α-h ydroxy derivative 22.
Th e vicin al couplin g con stan ts of com poun ds 26 an d 30 with keto group
in position 6 are close to th e con stan ts of th e ch air con form ation . Th e sam e
result was foun d by m olecular m odellin g. Con form ers with oth er th an
ch air con form ation h ave en ergy at least 7 kJ m ol–1 h igh er.
Addition al in form ation on th e A rin g con form ation of 3-oxo triterpen -
oids could be obtain ed in gen eral from NOE con tacts between sterically
close h ydrogen atom s. However th e in spection of m olecular m odels of four
above discussed con form ation s of rin g A sh owed, th at th ree con form ers –
C, T2 an d S – h ave sh ort in teratom ic distan ces (≤3 Å) on ly between th e
m eth yl group in position 25 an d h ydrogen s 1β an d 2β an d th us th e NOESY
spectra can n ot be used to distin guish between th ese con form ation s. Th e
correspon din g cross peaks were observed in NOESY spectra of com poun ds
12, 15, 30 an d 32. On th e oth er h an d, th e in teratom ic distan ce in th e con -
form ation T1 between th e 2β h ydrogen an d th e m eth yl group in position
25 is larger (close to 4 Å) but th e distan ce between 2α h ydrogen an d m eth yl
group 23 is sh orter (~3 Å). In th e NOESY spectrum of 6α-h ydroxy derivative
22, cross-peaks are observed between th e m eth yl group in position 23 an d
2α h ydrogen an d between m eth yl group 25 an d h ydrogen 1β but n ot 2β.
Th is observation is furth er eviden ce of th e T1 con form ation of th e 6α-
h ydroxy derivative.
FIG. 4
Rin gs A an d B of 6α-h ydroxylupan e derivatives (results of m olecular m odellin g)
Collect. Czech. Chem. Commun. 2006, Vol. 71, No. 8, pp. 1131–1160

1142
Dračínský, Hybelbauerová, Sejbal, Buděšínský:
EXPERIMENTAL
TLC was perform ed on silica gel plates G (Merck), visualisation by sprayin g with 10% sulfu-
ric acid an d h eatin g. Kieselgel 60 G (Merck) was used for preparative TLC (sprayin g with
0.3% m eth an olic solution of m orin ), colum n ch rom atograph y was carried out on silica gel
(Merck). Mixtures of h exan e an d eth er were used as eluen ts. Th e usual work-up m ean s dilu-
tion of reaction m ixture with water, extraction of products with eth er an d successive wash -
in g of eth er layer with dilute (1:4) h ydroch loric acid (if n ecessary), water, saturated sodium
h ydrogen carbon ate solution , water, dryin g over an h ydrous sodium sulfate an d evaporation
of solven ts un der reduced pressure. Sam ples for elem en tal an alysis were dried over ph osph o-
rus pen toxide un der reduced pressure. Startin g dih ydrobetulin 1 was prepared accordin g to
ref.51
Meltin g poin ts were determ in ed on a Kofler block an d are un corrected. Optical rotation s
were carried out in ch loroform (if n ot specified in experim en t), c 0.3–0.5, at am bien t tem -
perature on an autom atic polarim eter ETL-NPL (Ben dix–Ericsson ) with accuracy ±2 an d are
given in 10–1 deg cm 2 g–1. In frared spectra were recorded in ch loroform solution on a PE 684
(Perkin –Elm er) spectrom eter, waven um bers are given in cm –1. Mass spectra were recorded on
Fin n igan MAT-In cos 50 in strum en t, ion isin g electron en ergy 70 eV, direct in let tem peratures
150–180 °C. Data are presen ted in th e form at m/z (%), in ten sities are scaled to m ost in ten -
sive ion above m/z 80. Elem en tal an alyses were carried out on a Perkin –Elm er CHN An alyser
2400, Series II.
NMR spectra were recorded on in strum en t Varian UNITYINOVA 400 (1H at 400 MHz, 13C
at 100.5 MHz) in deuterioch loroform at 25 °C. For 1H NMR spectra, tetram eth ylsilan e (TMS)
was used as in tern al stan dard, ch em ical sh ifts (δ scale, ppm ) an d couplin g con stan ts (J, Hz)
were obtain ed by first order an alysis. 13C NMR ch em ical sh ifts were obtain ed from th e spec-
tra m easured in deuterioch loroform an d referen ced to solven t sign al (δ(CDCl3) 77.00). Th e
2D-H,H-COSY experim en ts were recorded in absolute value m ode usin g stan dard two-pulse
sequen ce. 2D-H,H-NOESY experim en ts were perform ed in ph ase sen sitive m ode with stan -
dard th ree-pulse sequen ce an d m ixin g tim e 0.3 s. Th e 2D-H,C-HSQC an d 2D-H,C-HMBC
m easurem en ts were perform ed as PFG experim en ts (HSQC in ph ase sen sitive an d HMBC in
absolute value m ode). All h eteron uclear 2D-H,C experim en ts were recorded with spectral
win dows 5000 Hz for proton an d 25 000 Hz for carbon .
Molecular m odellin g studies were perform ed usin g program m e Hyperch em 6 (Hypercube).
Th e con jugate gradien t m eth od for en ergy m in im isation s was used to con vergen ce (less th an
0.2 kJ m ol–1 RMS force). Force field MM+ was used for all com putation s. Gen eral protocol
for obtain in g lowest-en ergy con form ers by sim ulated an n ealin g: optim ised startin g structure
was subjected to dyn am ic run – 0.2 ps h eatin g from 300 to 1000 °C, 0.4 ps equilibration an d
1 ps coolin g to 200 °C followed by en ergy m in im isation . Every n ext run started from th e
previously m in im ised structure. A set of 100 structures was so obtain ed for each com poun d,
from wh ich th e lowest-en ergy con form ation was ch osen for evaluation .
28-Hydroxylupan -3-on e (3)
A solution of lupan e-3β,28-diol (1; 17 g, 38 m m ol) in pyridin e (340 m l) was cooled to 0 °C
an d th en brom in e (12 m l, 240 m m ol) was added. Th e m ixture was left stan din g at –24 °C
for th ree days an d th en it was diluted with water. Th e precipitate was filtered off, wash ed
with water an d dried at room tem perature. Th en th e precipitate was dissolved in eth er an d
th e solution was filtered th rough silica gel (30 g) an d worked up in th e usual m an n er.
Collect. Czech. Chem. Commun. 2006, Vol. 71, No. 8, pp. 1131–1160

B-Ring Substituted Lupane Derivatives
1143
Crystallisation from eth an ol afforded 28-h ydroxylupan -3-on e (3; 14.5 g, 85%). Th e ph ysical
con stan ts of th e com poun d were in agreem en t with th ose publish ed previously38
.
3-Oxolupan -28-yl Acetate (2)
To a solution of h ydroxyketon e 3 (3 g, 6.9 m m ol) in pyridin e (5 m l), acetic an h ydride (5 m l,
55 m m ol) was added an d th e m ixture was left stan din g at room tem perature overn igh t. Th e
m ixture was worked up in th e usual m an n er. Crystallisation from ch loroform –eth an ol gave
acetate 2 (2.9 g, 91%). Th e ph ysical data of th e com poun d were in agreem en t with th ose
publish ed previously38
.
28-Acetoxy-4,23-epoxy-3,4-secolupan e-3-n itrile (5)
A solution of 28-acetoxy-3,4-secolup-4(23)-en e-3-n itrile39 (4; 5.1 g, 10 m m ol) in ch loroform
(70 m l) was cooled to 0 °C. Th en 3-ch loroperoxyben zoic acid (60%, 4 g, 14 m m ol) was
added an d th e m ixture was left stan din g at 4 °C for 24 h . Th en eth er was added an d th e so-
lution was wash ed with 5% solution of potassium iodide an d th en with 5% solution of so-
dium sulfite to decolourise th e solution . Th is procedure was repeated un til th e potassium
iodide solution ch an ged colour to brown . Th en th e eth er layer was wash ed with water an d
worked up in th e usual m an n er. A m ixture of epoxy derivatives 5 (5.3 g, 100%) was ob-
tain ed. Th e m ixture was used with out purification in furth er reaction .
Reaction of Epoxysecon itrile 5 with Boron Trifluoride
A solution of crude m ixture of epoxy derivatives 5 (2.4 g, 4.8 m m ol) in dry toluen e (150 m l)
was deoxygen ated with low stream of argon an d th en boron trifluoride dieth yl eth erate
(7.5 m l, 68 m m ol) was added. Th e m ixture was refluxed un der argon for 10 h . After coolin g
to room tem perature th e reaction m ixture was worked up in th e usual m an n er. Evaporation
residue con tain ed (accordin g to TLC) two com pon en ts. Th e m ixture was separated by col-
um n ch rom atograph y on silica gel (250 g). A m ixture of eth er an d h exan e (1:10) was used as
eluen t. Followin g com poun ds were isolated (accordin g to in creasin g polarity).
22(17→28)Abeo-24-norlup-3-one (7): 130 m g (7%), m .p. 165–169 °C (ch loroform –eth an ol),
[α]D –9. IR: 1700 (C=O), 1453, 1207. EI-MS, m/z (%): 410 (M+, 22), 395 (1), 367 (3), 205
(100), 149 (44), 109 (77). 1H NMR: 0.677 d, 3 H (H-29, J(29,20) = 6.7); 0.919 d, 3 H (H-30,
J(30,20) = 6.5); 0.968 s, 3 H (H-26); 0.998 d, 3 H (H-23, J(23,4) = 6.5); 1.066 d, 3 H (H-25, J =
1.0); 1.115 s, 3 H (H-27); 1.91 dh , 1 H (H-20, J1 = 10.5, J2 = 6.5); 2.02 bm , 1 H (H-17, ΣJ =
25); 2.08 ddd, 1 H (H-1β, J(1β,1α) = 13.2, J(1β,2α) = 2.4, J(1β,2β) = 7.1); 2.28 m , 1 H (H-4β,
J(4β,5) = 11.8, J(4β,23) = 6.6); 2.33 ddd, 1 H (H-2α, J(2α,2β = 15.3, J(2α,1α) = 5.6, J(2α,1β) =
2.4); 2.36 bddd, 1 H (H-19, J1 = 10.6, J2 = 4.8, J3 = 2.6); 2.47 dddd, 1 H (H-2β, J(2β,1α) =
13.2, J(2β,2α) = 15.3, J(2β,1β) = 7.1, J(2β,4β) = 1.1); 2.72 bddd, 1 H (H-12β, J1 = 15.0, J2 = 5.0,
J3 = 2.4). For 13C NMR, see Table I. For C29H46O (410.7) calculated: 84.81% C, 11.29% H;
foun d: 84.32% C, 11.08% H.
3-Oxo-24-norlupan-28-yl acetate (6): 346 m g (15%), m .p. 130–131 °C (ch loroform –eth an ol),
[α]D 0. IR: 1244 (C–O–C), 1702 (C=O), 1727 (CH3COO). EI-MS, m/z (%): 470 (M+, 14), 410
(6), 397 (33), 367 (5), 204 (100), 191 (64), 43 (55). 1H NMR: 0.764 d, 3 H (H-29, J(29,20) =
6.7); 0.845 d, 3 H (H-30, J(30,20) = 6.8); 0.947 d, 3 H (H-27, J = 0.7); 0.982 d, 3 H (H-23,
J(23,4) = 6.6); 1.036 d, 3 H (H-25, J = 0.7); 1.104 s, 3 H (H-26); 2.067 s, 3 H (OAc); 2.02 ddd,
1 H (H-1β, J(1β,1α) = 13.1, J(1β,2β) = 7.0, J(1β,2α) = 2.3); 2.27 ddq, 1 H (H-4β, J(4β,5) = 12.0,
Collect. Czech. Chem. Commun. 2006, Vol. 71, No. 8, pp. 1131–1160

1144
Dračínský, Hybelbauerová, Sejbal, Buděšínský:
J(4β,23) = 6.6, J(4β,2β) = 1.0); 2.30 ddd, 1 H (H-2α, J(2α,2β) = 15.0, J(2α,1α) = 5.5, J(2α,1β) =
2.3); 2.44 dddd, 1 H (H-2β, J(2β,2α) = 15.0, J(2β,1α) = 13.4, J(2β,1β) = 7.0, J(4β,2β) = 1.1);
3.82 dd, 1 H (H-28a, J(28a,28b) = 11.0, Jl.r. = 1.4); 4.26 dd, 1 H (H-28b, J(28b,28a) = 11.0,
Jl.r. = 1.8). For 13C NMR, see Table I. For C31H50O3 (470.7) calculated: 79.10% C, 10.71% H;
foun d: 79.07% C, 10.94% H.
28-Ben zoyloxy-3,4-secolup-4(23)-en e-3-n itrile (8)
To th e solution of acetate 4 (10 g, 21 m m ol) in eth an ol (300 m l) a solution of sodium h y-
droxide (5 g) in eth an ol (150 m l) was added. Th e m ixture was refluxed for 1 h th en it was
cooled to room tem perature an d diluted with water (1 l). Th e precipitate was filtered off,
wash ed with water an d dried at 100 °C. 1H NMR spectrum of th e precipitate was in agree-
m en t with NMR data publish ed previously39 for 28-h ydroxy-3,4-secolup-4(23)-en e-3-n itrile.
Th e precipitate was dissolved in pyridin e (50 m l) an d th en ben zoyl ch loride (20 m l, 110 m m ol)
was added. Th e m ixture was left stan din g at room tem perature overn igh t an d th en it was
worked up in th e usual m an n er. Crystallisation from ch loroform –eth an ol gave 6.5 g (77%)
of th e title com poun d 8, m .p. 162–163 °C, [α]D –1. IR: 2249 (n itrile), 1714 (C=O), 1454,
1389, 1278, 1120. EI-MS, m/z (%): 543 (M+, 11), 421 (23), 408 (25), 378 (13), 340 (10), 229
(18), 191 (50), 135 (30), 105 (100). For 1H NMR, see Table IV. For 13C NMR, see Table I. For
C
37H53NO2 (543.8) calculated: 81.72% C, 9.82% H, 2.58% N; foun d: 81.67% C, 9.91% H,
2.35% N.
28-Ben zoyloxy-4,23-epoxy-3,4-secolupan e-3-n itrile (9)
Th e solution of com poun d 8 (10.3 g, 19 m m ol) in ch loroform (125 m l) was cooled to 0 °C.
Th en 3-ch loroperoxyben zoic acid (60%, 8 g, 28 m m ol) was added an d th e m ixture was left
stan din g at 4 °C for 16 h . Th en eth er was added an d th e solution was wash ed with 5% solu-
tion of potassium iodide an d th en with 5% solution of sodium sulfite to decolourise th e so-
lution . Th is procedure was repeated as lon g as th e potassium iodide solution ch an ged colour
to brown . Th en th e eth er layer was wash ed with water an d worked up in th e usual m an n er.
A m ixture of epoxy derivatives (10.6 g, 100%) was obtain ed. Multiple crystallisation gave
on e isom er, th e title com poun d 9, m .p. 179–181 °C, [α]D –1. IR: 2249 (n itrile), 1712 (C=O),
1454, 1277. EI-MS, m/z (%): 559 (M+, 2), 530 (2), 501 (2), 437 (9), 424 (11), 380 (7), 351 (4),
229 (18), 191 (32), 135 (28), 105 (100). 1H NMR: 0.791 d (J = 6.6), 0.866 d (J = 6.7); 0.952 s,
0.955 s, 1.109 s, 1.275 s , 6 × 3 H (6 × CH3); 2.63 d, 1 H (H-23a, J(23a,23b) = 4.4); 2.72 d,
1 H (H-23b, J(23b,23a) = 4.4); 4.05 d, 1 H (H-28a, J(28a,28b) = 11.1); 4.51 dd, 1 H (H-28b,
J(28b,28a) = 11.1, Jl.r. = 1.5); 8.05 m , 2 H, 7.56 m , 1 H an d 7.44 m , 2 H (C6H5). For
13C NMR, see Table I. For C37H53NO3 (559.8) calculated: 79.38% C, 9.54% H, 2.50% N;
foun d: 78.61% C, 9.63% H, 2.31% N.
3-Oxo-24-n orlupan -28-yl Ben zoate (10)
Th e solution of crude m ixture of isom eric epoxides 9 (5 g, 8.9 m m ol) in dry toluen e (200 m l)
was deoxygenated with low stream of argon and then boron trifluoride etherate (6 m l, 54 m m ol)
was added. Th e m ixture was refluxed un der argon for 2 h , th en
2 M h ydroch loric acid
(30 m l) was added an d th e m ixture was refluxed for an oth er 30 m in . After coolin g to room
tem perature th e reaction m ixture was worked up in th e usual m an n er. Th e crude product
was filtered th rough a colum n of silica gel (eluen t: ch loroform –petroleum eth er, 2:1).
Collect. Czech. Chem. Commun. 2006, Vol. 71, No. 8, pp. 1131–1160

B-Ring Substituted Lupane Derivatives
1145
TABLE I
13C NMR ch em ical sh ifts of com poun ds 6–10 an d 12–15 in CDCl3
Carbon
6
7
8
9
10
12
13
14
15
1
40.30
37.44
213.84
44.56
53.25
22.12
32.97
40.56
47.65
36.68
21.40
26.77a
37.10
42.93
26.82a
29.78
46.40
48.02
44.47
29.41
21.55
34.60
11.55
–
40.35
37.40
213.94
44.59
53.26
22.14
33.61
40.62
48.26
36.82
22.38
25.14
132.40
43.90
26.12
25.60
32.95
136.00
45.53
27.28
30.37
22.20
11.67
–
34.36
11.42
120.22
147.00
50.52
24.19
32.59
40.69
40.11
39.44
21.52
26.48
37.20
43.34
26.98
29.94
46.84
48.10
44.55
29.47
21.65
34.81
113.98
22.78
19.68
16.07
14.52
63.20
14.90
22.89
34.58
11.63
119.74
57.93
49.91
20.98
32.39
40.55
40.55
38.74
21.20
26.35
37.23
43.29
26.91
29.86
46.79
48.04
44.51
29.45
21.62
34.76
55.41
20.70
19.76
15.92
14.77
63.12
14.86
22.86
40.34
37.46
213.9
44.61
53.29
22.16
33.00
40.62
47.69
36.72
21.44
26.80
37.23
43.02
26.94
30.01
46.85
48.15
44.61
29.47
21.65
34.82
11.58
–
39.83
36.00
209.98
63.59
57.45
19.62
33.95
40.76
48.58
37.22
21.43
26.69
37.22
43.02
26.98
29.95
46.84
48.10
44.57
29.47
21.65
34.81
201.24
17.17
15.02
16.04
14.54
63.17
14.89
22.90
36.56
33.31
198.92
128.05
164.42
24.56
32.78
40.52
49.09
39.31
21.54
26.88
37.41
43.22
26.98
29.86
46.93
47.96
44.54
29.44
21.62
34.80
11.02
–
36.53
33.29
198.96
128.06
164.49
24.58
32.83
40.53
49.10
39.31
21.59
26.87
36.99
43.17
26.91
29.13
47.80
47.95
44.49
29.46
21.70
33.98
11.03
–
35.28
33.67
217.47
48.41
146.98
119.29
33.12
38.73
43.40
37.12
22.75
26.45
37.00
42.00
27.17
29.09
47.80
48.19
44.63
29.58
21.75
33.97
27.97
30.01
17.44
15.66
15.23
60.42
14.88
22.93
2
3
4
5
6
7
8
9
10
11
12
13
14
15
16
17
18
19
20
21
22
23
24
25
13.46
15.96
14.43
62.77
14.83
22.87
13.50
17.77
20.52
34.62
20.97
20.94
13.50
16.03
14.51
63.26
14.89
22.91
18.14
15.59
14.38
63.19
14.88
22.89
18.13
15.52
14.32
60.52
14.88
22.92
26
27
28
29
30
OAc or OBz
C=O
1′
171.70
21.03
–
–
–
–
–
166.94
130.50
129.53
128.35
132.84
166.91
130.44
129.53
128.83
132.84
166.97
130.53
129.54
128.36
132.84
166.94
130.48
129.54
128.36
132.87
166.95
130.47
129.53
128.36
132.86
–
–
–
–
–
–
–
–
–
–
2′, 6′
3′, 5′
4′
a
Sign als m ay be m utually in terch an ged.
Collect. Czech. Chem. Commun. 2006, Vol. 71, No. 8, pp. 1131–1160

1146
Dračínský, Hybelbauerová, Sejbal, Buděšínský:
Crystallisation from ch loroform –eth an ol gave th e title com poun d 10 (2.5 g, 53%), m .p.
196–198 °C, [α]D +5. IR: 1700 broad (2 × C=O), 1454, 1388, 1316, 1277, 1176, 1118. EI-MS,
m/z (%): 532 (M+, 10), 410 (16), 397 (26), 367 (7), 204 (60), 191 (40), 163 (25), 135 (29), 105
(100). For 1H NMR, see Table IV. For 13C NMR, see Table I. For C36H52O3 (532.8) calculated:
81.15% C, 9.84% H; foun d: 80.80% C, 9.85% H. Preparative TLC of m oth er liquor gave
25 m g of 3,24-dioxolupan -28-yl ben zoate (12), m .p. 225–229 °C, [α]D –18. IR: 2871 (CH=O),
1722, 1709 (2 × C=O), 1278, 1119. EI-MS, m/z (%): 532 (M+ – CO, 55), 425 (5), 410 (3), 397
(4), 229 (15), 191 (20), 135 (27), 105 (100). HRMS, m/z: 560.3879 (for C37H52O4 calculated:
560.3866). For 1H NMR, see Table IV. For 13C NMR, see Table I. Before addition of h ydro-
ch loric acid, th e reaction m ixture con tain ed also 3-oxo-23-n orlupan -28-yl ben zoate (11).
Th e attem pts to isolate th is com poun d failed because of easy isom erisation to 3-oxo-24-n or-
lupan -28-yl ben zoate (10). 1H NMR spectrum of 3-oxo-23-n orlupan -28-yl ben zoate (11) was
obtain ed by elim in ation of sign als of com poun d 10 from a spectrum of th e reaction m ix-
ture. 1H NMR: 0.791 d, 3 H (H-29, J(29,20) = 6.7); 0.864 d, 3 H (H-30, J(30,20) = 6.8);
0.988 s, 3 H (H-27); 1.021 s, 3 H (H-25); 1.105 d, 3 H (H-24, J(24,4α) = 7.8); 1.133 s, 3 H
(H-26); 2.58 ddd, 1 H (H-2β, J(2β,2α) = 15.7, J(2β,1α) = 12.7, J(2β,1β) = 6.9); 4.07 d, 1 H
(H-28a, J(28a,28b) = 11.0); 4.53 d, 1 H (H-28b, J(28b,28a) = 11.0); 8.05 m , 2 H, 7.56 m , 1 H
an d 7.44 m , 2 H (C6H5).
3-Oxo-24-n orlup-4-en -28-yl Ben zoate (13)
Keton e 10 (7.7 g, 14.4 m m ol) was dissolved in warm acetic acid (550 m l). Th e solution was
cooled to th e room tem perature an d th en 33% solution of h ydrogen brom ide in acetic acid
(0.3 m l) was added. Th e m ixture was con tin uously stirred an d, durin g 30 m in , brom in e
(2.56 g, 16 m m ol) in acetic acid (64 m l) was added dropwise. Th en 33% solution of h ydro-
gen brom ide in acetic acid (70 m l, 288 m m ol) was added an d th e m ixture was left stan din g
at room tem perature for two days in th e dark. Th e reaction m ixture was th en diluted with
water, th e precipitate was filtered off, wash ed with saturated solution of sodium h ydrogen -
carbon ate an d with water an d th en it was dried on air at room tem perature. Precipitate (7.7 g)
of th e title com poun d 13 was obtain ed, pure en ough for use for furth er reaction . A sam ple
for an alysis was obtain ed by preparative TLC (eluen t: eth er–h exan e, 1:2). M.p. 212–217 °C
(ch loroform –eth an ol), [α]D +47. IR: 1713 (C=O), 1655 (C=O), 1604 (C=C), 1453, 1388, 1315,
1278, 1117. EI-MS, m/z (%): 530 (M+, 87), 365 (8), 351 (5), 229 (25), 191 (27), 147 (18), 138
(36), 105 (100). 1H NMR: 0.793 d, 3 H (H-29, J(29,20) = 6.7); 0.873 d, 3 H (H-30, J(30,20) =
6.7); 0.953 s, 3 H (H-27); 1.144 s, 3 H (H-25); 1.284 s, 3 H (H-26); 1.791 d, 3 H (H-23, Jl.r.
=
1.1); 2.01 ddd, 1 H (H-1β, J(1β,1α) = 13.1, J(1β,2β) = 5.1, J(1β,2α) = 2.8); 2.35 ddd, 1 H
(H-2α, J(2α,2β) = 17.0, J(2α,1α) = 4.7, J(2α,1β) = 2.7); 2.46 ddd, 1 H (H-2β, J(2β,2α) = 17.0,
J(2β,1α) = 15.0, J(2β,1β) = 5.2); 2.64 ddd, 1 H (H-6a, J(6a,6b) = 14.8, J(6a,7a) = 3.4, J(6a,7b) =
3.4); 4.07 dd, 1 H (H-28a, J(28a,28b) = 11.1, Jl.r. = 1.2); 4.56 dd, 1 H (H-28b, J(28b,28a) =
11.1, Jl.r. = 1.8); 7.45 m , 2 H, 7.57 m , 1 H an d 8.06 m , 2 H (C6H5). For 13C NMR, see Table I.
For C36H50O3 (530.8) calculated: 81.46% C, 9.49% H; foun d: 81.25% C, 9.48% H.
28-Hydroxy-24-n orlup-4-en -3-on e (14)
To th e solution of ben zoate 13 (3.5 g, 6.6 m m ol) in eth an ol (100 m l) a solution of potas-
sium h ydroxide (1.7 g, 30 m m ol) in eth an ol (50 m l) was added. Th e m ixture was refluxed
for 1 h an d th en it was cooled to room tem perature, diluted with water (300 m l) an d ex-
tracted with eth er. Th e eth er layer was worked up in th e usual m an n er. Th e evaporation res-
Collect. Czech. Chem. Commun. 2006, Vol. 71, No. 8, pp. 1131–1160

B-Ring Substituted Lupane Derivatives
1147
idue (2.8 g) was used for an oth er reaction with out furth er purification . A sam ple for an alysis
was obtain ed by colum n ch rom atograph y (see th e n ext experim en t) an d crystallisation from
ch loroform –eth an ol. M.p. 245–249 °C, [α]D +124. IR: 3625 (OH), 1656 (C=O), 1607 (C=C),
1458, 1376, 1329, 1018. For 1H NMR, see Table IV. For 13C NMR, see Table I. For C29H46O2
(426.7) calculated: 81.63% C, 10.87% H; foun d: 81.12% C, 10.45% H.
28-Hydroxylup-5-en -3-on e (15)
Potassium (220 m g, 5.6 m m ol) was dissolved in warm tert-butyl alcoh ol (10 m l). To 7.5 m l
of th is solution keton e 14 (250 m g, 0.59 m m ol) was added. Th is m ixture was refluxed un der
argon for 1 h . Th en m eth yl iodide (100 µl, 1.6 m m ol) was added an d th e m ixture was
refluxed an oth er 15 m in an d th en it was left stan din g at room tem perature for 20 m in .
Th en water (30 m l) was added an d th e m ixture was extracted with eth er. Th e eth er layer
was worked up in th e usual m an n er. Th e evaporation residue (240 m g) con tain ed (accordin g
to TLC) four com pon en ts. Th e m ixture was separated by colum n ch rom atograph y on silica
gel (25 g). Eth er–h exan e (1:1) was used as eluen t. Th e followin g com poun ds were obtain ed
an d th ey were crystallised from ch loroform –eth an ol (accordin g to in creasin g polarity).
28-Methoxylup-5-en-3-one (16): 15 m g (5.6%), m .p. 185–191 °C, [α]D +15. IR: 1702 (C=O),
1666 (C=C), 1462, 1384, 1371, 1100, 1024, 966, 816. EI-MS, m/z (%): 454 (M+, 29), 422 (4),
409 (23), 397 (8), 229 (21), 205 (36), 203 (35), 191 (52), 124 (45), 107 (100). For 1H NMR,
see Table V. For 13C NMR, see Table II.
28-Methoxy-24-norlup-4-en-3-one (17): 14 m g (5.4%), m .p. 218–221 °C, [α]D +54. IR: 1655
(C=O), 1606 (C=C), 1458, 1376, 1100, 963. EI-MS, m/z (%): 440 (M+, 100), 408 (36), 395
(13), 366 (18), 271 (20), 229 (23), 203 (33), 191 (62), 161 (36), 147 (47), 138 (49), 121 (56).
HRMS, m/z: 440.3659 (for C30H48O2 calculated: 440.3654). 1H NMR: 0.763 d, 3 H (H-29,
J(29,20) = 6.7); 0.854 d, 3 H (H-30, J(30,20) = 6.8); 0.919 s, 3 H (H-27); 1.137 s, 3 H (H-25);
1.253 s, 3 H (H-26); 1.793 d, 3 H (H-23, Jl.r.= 1.2); 2.00 ddd, 1 H (H-1β, J(1β,1α) = 13.1,
J(1β,2β) = 5.2, J(1β,2α) = 2.8); 2.34 ddd, 1 H (H-2α, J(2α,2β) = 17.1, J(2α,1α) = 4.9, J(2α,1β) =
2.6); 2.45 ddd, 1 H (H-2β, J(2β,2α) = 16.9, J(2β,1α) = 15.0, J(2β,1β) = 5.2); 2.64 ddd, 1 H
(H-6a, J(6a,6b) = 14.8, J(6a,7a) = 3.5, J(6a,7b) = 3.5); 3.06 dd, 1 H (H-28a, J(28a,28b) = 9.2,
Jl.r. = 1.1); 3.49 dd, 1 H (H-28b, J(28b,28a) = 9.2, Jl.r. = 1.6); 3.35 s, 3 H (OCH3). For 13C NMR,
see Table II.
28-Hydroxylup-5-en-3-one (15): 88 m g (34%), m .p. 188–192 °C, [α]D +22. IR: 3626 (OH),
1701 (C=O), 1666 (C=C), 1462, 1385, 1371, 1111, 1025, 1017. EI-MS, m/z (%): 440 (M+, 72),
422 (8), 409 (15), 247 (15), 217 (18), 203 (21), 191 (22), 177 (20), 152 (38), 133 (28), 124
(100). For 1H NMR, see Table V. For 13C NMR, see Table I. For C30H48O2 (440.7) calculated:
81.76% C, 10.98% H; foun d: 80.21% C, 11.33% H.
28-Hydroxy-24-norlup-4-en-3-one (14): 94 m g (38%), iden tical with sam ple obtain ed by
alkalin e h ydrolysis of ben zoate 13 (see previous experim en t).
Lup-5-en e-3β,28-diol (18)
To a solution of keton e 15 (200 m g, 0.45 m m ol) in a m ixture of ben zen e (5 m l) an d m eth a-
n ol (7 m l) sodium boroh ydride (200 m g, 5.3 m m ol) was added durin g 15 m in an d th e m ix-
ture was left stan din g at room tem perature. Th en 2 M h ydroch loric acid (10 m l) was added
dropwise an d th e m ixture was extracted with eth er, th e eth er layer was worked up in th e
usual m an n er. Crystallisation from ch loroform –eth an ol gave title com poun d 18 (162 m g,
Collect. Czech. Chem. Commun. 2006, Vol. 71, No. 8, pp. 1131–1160

1148
Dračínský, Hybelbauerová, Sejbal, Buděšínský:
TABLE II
13C NMR ch em ical sh ifts of com poun ds 16–22 in CDCl3
Carbon
16
17
18
19a
20
21b
22
1
2
35.27
33.68
217.50
48.42
146.96
119.36
33.13
38.73
43.41
37.13
22.80
26.47
37.22
41.97
27.36
29.85
47.19
48.34
44.81
29.60
21.94
34.72
27.97
30.01
17.45
15.67
15.27
71.19
14.91
22.96
59.66
36.54
33.31
198.94
128.05
164.53
24.60
32.82
40.54
49.10
39.31
21.62
26.93
37.20
43.16
27.07
29.90
47.35
47.97
44.67
29.49
21.88
34.75
11.03
–
38.99
27.41
78.09
42.01
146.08
119.92
34.40
38.13
46.22
37.25
22.24
26.51
37.01
42.27
27.21
29.11
47.81
48.27
44.68
29.58
21.76
33.96
27.96
20.28
22.80
16.63
15.48
60.47
14.89
22.94
–
38.66
23.83
79.91
40.81
145.35
120.51
34.40
38.12
46.16
37.19
22.15
26.46
37.30
42.28
27.20
29.65
46.35
48.31
44.60
29.52
21.62
34.57
27.86
20.44
24.04
16.74
15.43
62.72
14.88
22.89
–
38.46
26.71
78.68
39.26
60.48
68.75
46.72
42.90
49.52
39.15
20.72
26.93
36.34
42.23
26.93
29.23
47.88
48.00
44.51
29.44
21.68
33.96
30.88
15.46
17.06
17.48
14.63
60.48
14.85
22.91
–
37.98
23.20
80.41
39.38
58.15
70.77
41.92
42.07
49.43
37.60
20.66
26.56
36.63
42.98
26.96
29.71
46.43
48.03
44.47
29.40
21.56
34.60
30.34
16.74
17.05
17.27
14.60
62.70
14.86
22.87
–
39.56
33.02
219.96
47.15
58.56
67.82
44.48
42.89
47.93
38.19
21.71
26.80
36.51
41.70
26.91
29.14
47.88
48.43
44.48
29.48
21.71
33.97
31.90
19.59
17.62
16.39
14.57
60.45
14.86
22.91
–
3
4
5
6
7
8
9
10
11
12
13
14
15
16
17
18
19
20
21
22
23
24
25
26
27
28
29
30
OMe
18.14
15.52
14.36
71.34
14.89
22.95
59.68
a
Sign als of acetate in th e position 3 (δ 170.83 an d 21.33), an d 28 (δ 171.62 an d 21.05);
sign als of acetate in th e position 3 (δ 170.01 an d 21.28), 6 (δ 171.05 an d 21.96) an d 28
b
(δ 171.58 an d 21.00).
Collect. Czech. Chem. Commun. 2006, Vol. 71, No. 8, pp. 1131–1160

B-Ring Substituted Lupane Derivatives
1149
81%), m .p. 269–271 °C, [α]D –22. IR: 3624 (OH), 1462, 1385, 1020. EI-MS, m/z (%): 442 (M+,
36), 424 (20), 359 (89), 203 (26), 187 (100), 175 (42), 161 (30), 147 (38), 133 (68), 119 (72).
For 1H NMR, see Table V. For 13C NMR, see Table II. For C30H50O2 (442.7) calculated:
81.39% C, 11.38% H; foun d: 80.94% C, 11.50% H.
Lup-5-en e-3β,28-diyl Diacetate (19)
To a solution of diol 18 (10 m g, 0.023 m m ol) in pyridin e (0.4 m l), acetic an h ydride (0.4 m l,
4.4 m m ol) was added an d th e m ixture was left stan din g at room tem perature overn igh t. Th e
m ixture was worked up in th e usual m an n er. Crystallisation from ch loroform –eth an ol gave
diacetate 19 (10 m g, 84%), m .p. 230–235 °C, [α]D –8. IR: 1724 (C=O), 1460, 1387, 1367,
1252, 1031, 982. EI-MS, m/z (%): 526 (M+, 5), 466 (18), 451 (15), 398 (18), 325 (6), 302 (4),
187 (100), 175 (41). 1H NMR: 0.774 d, 3 H (H-29, J(29,20) = 7.2); 0.850 d, 3 H (H-30,
J(30,20) = 6.8); 0.941 s, 3 H (H-27); 1.054 s, 3 H (H-26); 1.151 s, 3 H (H-24); 1.031 s, 1.103 s,
2 × 3 H (H-23 an d H-25); 2.057 s, 2.064 s, 2 × 3 H (2 × OAc); 3.85 dd, 1 H (H-28a,
J(28a,28b) = 11.1, Jl.r. = 1.2); 4.25 dd, 1 H (H-28b, J(28b,28a) = 11.1, Jl.r. = 1.8); 4.46 m , 1 H
(H-3, ΣJ = 16.5); 5.57 dd, 1 H (H-6, J(6,7a) = 3.1, J(6,7b) = 5.0). For 13C NMR, see Table II.
For C34H54O4 (526.8) calculated: 77.52% C, 10.33% H; foun d: 76.95% C, 10.46% H.
Lupan e-3β,6α,28-triol (20)
Olefin 18 (100 m g, 0.23 m m ol) was placed in to a 3-m l reaction flask with Teflon cap an d
1 M solution of BH3 in tetrah ydrofuran (2.5 m l, 2.5 m m ol) was added. Th e flask was covered
with m etal n et an d it was warm ed to 80 °C in water bath for 8 h an d th en it was left stan d-
in g at room tem perature overn igh t. Th en a solution of sodium h ydroxide (0.4 g, 10 m m ol)
in eth an ol (10 m l) an d 30% h ydrogen peroxide (6 m l, 58 m m ol) was added. Th e m ixture
was stirred for 4 h an d th en it was extracted with eth er. Th e eth er layer was wash ed succes-
sively with water, 2 M h ydroch loric acid, 5% solution of potassium iodide an d with 5% solu-
tion of sodium sulfite to decolourise th e solution . Th is procedure was repeated un til th e
potassium iodide solution turn ed to brown . Th en th e eth er layer was wash ed with water,
dried over an h ydrous sodium sulfate an d evaporated un der reduced pressure. Preparative
TLC (eluen t: h exan e–eth er, 1:3) an d crystallisation from ch loroform –eth an ol gave two com -
poun ds: startin g com poun d 18 (22 m g, 22%) an d triol 20 (58 m g, 56%), m .p. 262–265 °C,
[α]D 0. IR: 3625 (OH), 1463, 1386, 979. EI-MS, m/z (%): 460 (M+, 1), 442 (5,5), 429 (6), 411
(17), 393 (3), 207 (25), 205 (20), 191 (22), 187 (100), 177 (25). HRMS, m/z: 460.3926 (for
C
30H52O3 calculated: 460.3916). For 1H NMR, see Table V. For 13C NMR, see Table II.
Lupan e-3β,6α,28-triyl Triacetate (21)
To a solution of triol 20 (10 m g, 0.022 m m ol) in dry pyridin e (0.4 m l) acetic an h ydride
(0.4 m l, 4.4 m m ol) was added an d th e m ixture was left stan din g at room tem perature over-
n igh t. Th en th e m ixture was worked up in th e usual m an n er. Crystallisation from m eth an ol
gave triacetate 21 (10 m g, 78%), m .p. 175–179 °C. IR: 1726 (C=O), 1460, 1368, 1255, 1030,
980. 1H NMR: 0.766 d, 3 H (H-29, J(29,20) = 6.8); 0.839 d, 3 H (H-30, J(30,20) = 6.8); 0.904
s, 3 H (H-25); 0.971 s, 0.975 s, 2 × 3 H (H-24 an d H-27); 1.028 s, 3 H (H-23); 1.182 s, 3 H
(H-26); 2.021 s, 3 H (OAc); 2.052 s, 6 H (2 × OAc); 3.83 d, 1 H (H-28a, J(28a,28b) = 11.0);
4.19 dd, 1 H (H-28b, J(28b,28a) = 11.0, Jl.r. = 1.7); 4.45 m , 1 H (H-3, ΣJ = 16.5); 5.29 td, 1 H
(H-6β, J(6β,5α) = 10.9, J(6β,7α) = 10.9, J(6β,7β) = 4.1). For 13C NMR, see Table II.
Collect. Czech. Chem. Commun. 2006, Vol. 71, No. 8, pp. 1131–1160

1150
Dračínský, Hybelbauerová, Sejbal, Buděšínský:
6α,28-Dih ydroxylupan -3-on e (22)
To a solution of triol 20 (13 m g, 0.028 m m ol) in pyridin e (0.5 m l) brom in e (20 µl, 0.40 m m ol)
was added an d th e m ixture was left stan din g at –24 °C for 3 days. Th en th e m ixture was di-
luted with water an d extracted with eth er. Th e eth er layer was wash ed with water, 2 M h y-
droch loric acid, 5% solution of sodium sulfite an d with saturated solution of sodium
h ydrogen carbon ate an d th en it was dried over an h ydrous sodium sulfate an d evaporated un -
der reduced pressure. Keton e 22 (10 m g, 77%) was obtain ed, am orph ous, [α]D +42. IR: 3620
(OH), 1700 (C=O), 1460, 1384, 1368, 1027. EI-MS, m/z (%): 458 (M+, 4), 440 (5), 427 (29),
409 (40), 203 (100), 191 (55), 177 (40), 149 (91). For 1H NMR, see Table V. For 13C NMR, see
Table II. For C30H50O3 (458.7) calculated: 78.55% C, 10.99% H; foun d: 78.21% C, 11.13% H.
3β,6α-Dih ydroxylupan -28-yl Pivalate (23)
A solution of triol 20 (70 m g, 0.15 m m ol) in dry pyridin e (1.8 m l) was cooled to –10 °C an d
th en pivaloyl ch loride (250 µl, 2.1 m m ol) was added. Th e m ixture was left stan din g at
–10 °C for 2 h , th en m eth an ol (20 m l) was added (to rem ove excess of pivaloyl ch loride)
an d th e m ixture was left stan din g at 4 °C overn igh t. Th e m ixture was extracted with eth er,
th e eth er layer was wash ed with 2 M h ydroch loric acid, water an d saturated solution of so-
dium h ydrogen carbon ate, th en it was dried with an h ydrous sodium sulfate an d evaporated
in vacuum . Evaporation residue (83 m g) was accordin g to TLC a m ixture of th ree com -
poun ds. Th e m ixture was separated by preparative TLC usin g eth er–h exan e (2:1) as eluen t.
Th e followin g com poun ds were obtain ed (accordin g to in creasin g polarity).
6α-Hydroxylupane-3β,28-diyl dipivalate (24): 13 m g, 14%, m .p. 295–297 °C (m eth an ol).
1H NMR: 0.770 d, 3 H (H-29, J(29,20) = 6.8); 0.840 d, 3 H (H-30, J(30,20) = 6.8); 0.981 s, 3 H
(H-27); 1.132 s, 3 H (H-26); 0.925 s, 1.081 s, 1.165 s, 3 × 3 H (H-23, H-24 an d H-25); 1.206 s,
18 H (2 × pivalate); 3.76 dd, 1 H (H-28a, J(28a,28b) = 11.0, Jl.r. = 1.0); 4.25 dd, 1 H (H-28b,
J(28b,28a) = 11.0, Jl.r. = 1.8); 4.05 td, 1 H (H-6β, J(6β,5α) = 11, J(6β,7α) = 11, J(6β,7β) = 3.2);
4.40 m , 1 H (H-3, ΣJ = 16.3). For 13C NMR, see Table III.
3β-Hydroxylupane-6α,28-diyl dipivalate (25): 6 m g, 6%, am orph ous, [α]D 0. 1H NMR: 0.765 d,
3 H (H-29, J(29,20) = 6.4); 0.839 d, 3 H (H-30, J(30,20) = 6.8); 0.824 s, 0.961 s, 0.978 s, 1.187 s,
1.228 s, 5 × 3 H (5 × CH3); 1.198 s, 1.206 s, 2 × 9 H (2 × pivalate); 3.19 m , 1 H (H-3, ΣJ =
16.5); 3.80 dd, 1 H (H-28a, J(28a,28b) = 11.4, Jl.r. = 1.0); 4.17 dd, 1 H (H-28b, J(28b,28a) =
11.4, Jl.r. = 1.8); 5.32 td, 1 H (H-6β, J(6β,5α) = 10.9, J(6β,7α) = 10.9, J(6β,7β) = 3.9). For
13C NMR, see Table III.
3β,6α-Dihydroxylupan-28-yl pivalate (23): 44 m g, 53%, m .p. 125–129 °C (m eth an ol), [α]D
–11. IR: 3608 (OH), 1716 (C=O), 1480, 1462, 1387, 1367, 1174, 1033. EI-MS, m/z (%): 544
(M+, 1), 526 (3), 509 (4), 493 (12), 411 (6), 205 (22), 191 (30), 187 (100). HRMS, m/z:
544.4471 (for C35H60O4 calculated: 544.4492). 1H NMR: 0.766 d, 3 H (H-29, J(29,20) = 6.8);
0.840 d, 3 H (H-30, J(30,20) = 6.8); 0.894 s, 3 H (H-25); 0.979 s, 0.982 s, 2 × 3 H (H-24 an d
H-27); 1.128 s, 3 H (H-26); 1.314 s, 3 H (H-23); 1.206 s, 9 H (pivalate); 3.18 m , 1 H (H-3,
ΣJ = 16.3); 3.77 dd, 1 H (H-28a, J(28a,28b) = 11.1, Jl.r. = 1.8); 4.24 dd, 1 H (H-28b,
J(28b,28a) = 11.1, Jl.r. = 1.3); 4.05 td, 1 H (H-6β, J(6β,5α) = 10.6, J(6β,7α) = 10.6, J(6β,7β) =
3.8). For 13C NMR, see Table III.
Collect. Czech. Chem. Commun. 2006, Vol. 71, No. 8, pp. 1131–1160

B-Ring Substituted Lupane Derivatives
1151
TABLE III
13C NMR ch em ical sh ifts of com poun ds 23–26 an d 28–33 in CDCl3
Carbon
23a
24b
25c
26d
28e
29f
30
31
32
33g
1
2
38.48
26.95
78.71
39.28
60.48
68.74
46.69
42.92
49.52
39.15
20.68
26.68
36.69
42.19
26.95
29.88
46.78
48.11
44.54
29.41
21.57
34.63
30.90
15.48
17.07
17.57
14.66
62.44
14.87
22.88
38.04
23.11
80.19
39.01
60.56
68.54
46.72
42.93
49.42
39.16
20.72
26.65
36.69
42.17
26.95
29.86
46.78
48.11
44.55
29.41
21.58
34.63
30.76
16.70
17.11
17.58
14.67
62.45
14.87
22.87
38.45
26.84
78.73
38.57h
58.08
71.58
42.05
42.98
49.57
39.76h
20.64
26.60
36.64
42.10
26.93
29.68
46.76
48.08
44.51
29.40
21.55
34.57
30.82
15.49
16.96
17.18
14.64
62.34
14.87
22.86
40.98
33.80
214.56
46.90
65.19
211.19
51.93
47.61
49.99
43.46
21.29
26.26
36.93
43.31
26.92
29.70
46.68
48.24
44.52
29.42
21.58
34.54
24.10
21.67
16.34
16.07
14.94
62.16
14.84
22.84
40.71
27.59
79.14
39.66
55.60
68.89
42.17
40.04
50.70
36.66
20.92
26.95
36.30
43.10
26.94
29.84
46.46
48.22
44.52
29.47
21.60
34.66
27.64
16.89
17.66
16.99
14.94
62.79
14.94
22.93
40.37
23.89
80.88
38.65
55.61
68.75
42.22
40.01
50.59
36.63
20.90
26.98
36.23
43.06
26.87
29.79
46.44
48.19
44.59
29.42
21.57
34.62
27.57
18.13
17.66
16.96
14.89
62.75
14.86
22.91
41.01
33.81
214.54
46.93
65.20
211.38
52.06
47.72
50.03
43.51
21.38
26.34
36.64
43.35
26.92
29.12
47.82
48.18
44.55
29.50
21.70
33.93
24.14
21.70
16.37
16.05
14.94
60.40
14.86
22.91
34.50
27.46
75.21
38.12
68.47
54.85
32.79
36.98
41.06
41.06
20.17
26.63
36.78
43.37
26.90
29.05
47.74
48.26
44.42
29.41
21.80
33.97
21.57
19.89
20.11h
20.32h
14.99
60.63
14.85
22.92
34.85
33.53
217.5
47.77h
68.69
55.36
32.37
38.26
39.94
36.20
21.51i
26.40
36.82
42.69
26.90
29.02
47.66h
48.29
44.54
29.52
21.77i
33.94
23.03
24.32
19.55
19.15
15.33
60.49
14.85
22.92
37.95
23.45h
78.53
40.38
65.55
61.33
31.68
39.10
47.15
37.19
23.51h
26.81
36.64
43.00
27.37
29.61
46.32
48.23
44.50
29.48
21.59
34.53
24.57
20.11
18.65
16.85
15.52
62.63
14.90
22.87
3
4
5
6
7
8
9
10
11
12
13
14
15
16
17
18
19
20
21
22
23
24
25
26
27
28
29
30
a
b
28-OPiv: 178.91, 36.69, 27.28; 3-OPiv: 178.21, 38.32, 27.20, 28-OPiv: 178.92, 36.69,
c
d
27.20; 6-OPiv: 178.11, 27.17, 28-OPiv: 178.92, 36.64, 27.27;
27.24; 28-OAc: 171.65, 21.07; 3-OAc: 171.02, 21.33, 28-OAc: 171.60, 21.04; 3-OAc:
170.80, 21.26, 28-OAc: 171.61, 21.05;
terch an ged.
28-OPiv: 178.83, 36.93,
e
f
g
h,i
sign als with iden tical sym bols m ay be m utually in -
Collect. Czech. Chem. Commun. 2006, Vol. 71, No. 8, pp. 1131–1160

1152
Dračínský, Hybelbauerová, Sejbal, Buděšínský:
TABLE IV
1H NMR ch em ical sh ifts an d couplin g con stan ts of com poun ds 8, 10, 12 an d 14 in CDCl3
Hydrogen
8
10
12
14
H-1α
H-1β
1.68 m
1.68
1.29 m
1.36 m
1.57 m
2.03 ddd
(13.1; 7.0; 2.5)
2.33 m
2.06 ddd
(13.3; 7.0; 3.0)
2.43 ddd
(15.5; 5.9; 3.0)
2.64 ddd
(15.5; 12.7; 7.0)
–
2.00 ddd
(13.1; 5.2; 2.7)
2.34 ddd
(17.1; 4.9; 2.7)
2.45 ddd
(17.1; 14.8; 5.2)
–
H-2α
H-2β
2.20 m a
2.32 m a
2.45 td
(13.9; 13.9; 7.2)
–
2.27 m
1.08 m
1.41 m
1.61 m
H-3α
H-4β
H-5α
H-6a
H-6b
–
–
–
–
–
1.90 m
1.38 m
1.81 m
1.44 m
1.74 m
1.79 m
2.26 m
2.64 dt
(14.8; 3.5; 3.5)
1.56 m
1.56 m
1.56 m
1.29 m
1.58 m
1.21 m
1.66 m
1.74 m
1.08 m
1.76 m
1.20 m
1.93 ddd
(13.3; 4.6; 2.4)
1.40 m
1.75 m
1.87 m
1.51 m
1.58 m
0.85 m
1.84 m
1.79 d (1.2)
–
–
1.133 s
1.235 s
0.930 s
H-7a
H-7b
H-9
1.38 m
1.41 m
1.44 m
1.53 m
1.70 m
1.28 m
1.66 m
1.78 m
1.06 m
1.79 m
1.29 m
1.95 m
1.41m
1.41m
1.46 m
1.51 m
1.41 m
1.38 m
1.41 m
1.25 m
1.67 m
1.79 m
1.06 m
1.80 m
1.30 m
1.96 m
1.42 m
1.40 m
1.53 m
1.26 m
1.67 m
1.80 m
1.06 m
1.80 m
1.30 m
1.98 m
H-11a
H-11b
H-12a
H-12b
H-13
H-15a
H-15b
H-16a
H-16b
H-18
H-19
H-20
H-21a
H-21b
H-22a
H-22b
H-23a
H-23b
H-24
1.43 m
1.84 m
1.90 m
1.38 m
1.44 m
0.96 m
1.91 m
4.64 d (2.1)
4.88 m
1.724 s
0.861 s
1.126 s
1.001 s
1.42 m
1.85 m
1.91 m
1.53 m
1.69 m
0.95 m
1.92 m
0.98 d (5.5)
–
1.43 m
1.85 m
1.89 m
1.52 m
1.71 m
0.96 m
1.91 m
1.249 s
–
9.713 s
0.995 s
1.137 s
0.985 s
–
H-25
H-26
H-27
1.048 d (0.6)
1.141 s
0.975 s
H-28a
H-28b
H-29
4.06 dd (11.1; 1.4)
4.52 dd (11.1; 1.7)
0.870 d (6.8)
0.802 d (6.8)
4.06 dd (11.1; 1.4)
4.54 dd (11.1; 1.8)
0.791 d (6.7)
0.867 d (6.9)
4.06 dd (11.1; 1.2) 3.34 d (11.2)
4.53 dd (11.1; 1.7) 3.80 dd (11.2; 1.6)
0.792 d (6.6)
0.868 d (6.9)
0.770 d (6.9)
0.854 d (6.9)
H-30
OBz:
H-2′,6′
H-3′,5′
H-4′
8.05 m
7.44 m
7.56 m
8.05 m
7.45 m
7.56 m
8.05 m
7.45 m
7.56 m
–
–
–
a
Sign als m ay be m utually in terch an ged.
Collect. Czech. Chem. Commun. 2006, Vol. 71, No. 8, pp. 1131–1160

B-Ring Substituted Lupane Derivatives
1153
TABLE V
1H NMR ch em ical sh ifts an d couplin g con stan ts of com poun ds 15, 16, 18, 20 an d 22 in
CDCl3
Hydrogen 15
16
18
20
22
H-1α
H-1β
H-2α
1.56 m
1.97 m
1.56 m
1.96 m
0.99 m
1.78 m
1.68 m
0.95 m
1.68 m
1.22 m
1.63 m
1.82 m
2.54 ddd
(18.9; 9.2; 1.6)
2.43 ddd
(18.9; 10.2; 8.7)
–
2.53 ddd
(18.9; 9.3; 1.8)
2.43 ddd
(18.9; 10.4; 8.7)
–
2.71 ddd
(15.3; 12.0; 6.6)
2.26 ddd
(15.3; 9.8; 3.6)
–
H-2β
H-3α
1.68 m
1.62 m
3.21 m
(ΣJ = 16.2)
–
3.18 m
(ΣJ = 16.4)
0.86 m
H-5α
–
–
1.66 m
H-6
5.54 dd
(6.0; 2.4)
1.73 m
2.20 bd
(17.2)
5.55 dd
(5.6; 2.4)
1.72 m
2.20 bd
(16.8)
5.5710 dd
(5.2; 3.2)
1.68m
2.23 bd
(18.4)
4.05 td
(10.6; 10.6; 4.0)
1.47 m
3.93 td
(10.9; 10.9; 4.3)
1.50 m
H-7a
H-7b
1.69 m
1.79 m
H-9
1.63 m
1.44 m
1.54 m
1.25 m
1.66 m
1.68 m
1.05 m
1.83 m
1.19 m
1.93 m
1.62 m
1.44 m
1.56 m
1.26 m
1.66 m
1.68 m
1.04 m
1.71 m
1.15 m
1.91 m
1.42 m
1.46 m
1.54 m
1.23 m
1.63 m
1.62 m
1.03 m
1.77 m
1.17 m
1.88 m
1.30 m
1.26 m
1.50 m
1.26 m
1.86 m
1.63 m
1.03 m
1.76 m
1.19 m
1.91 m
1.40 m
1.25 m
1.48 m
1.25 m
1.63 m
1.65 m
1.08 m
1.76 m
1.19 m
1.93 ddd
(13.3; 4.6; 2.4)
1.47 m
1.72 m
1.86 m
1.48 m
1.62 m
0.86 m
1.84 m
1.335 s
1.319 s
0.75 d
H-11a
H-11b
H-12a
H-12b
H-13
H-15a
H-15b
H-16a
H-16b
H-18
H-19
H-20
H-21a
H-21b
H-22a
H-22b
H-23
H-24
H-25
1.40 m
1.73 m
1.89 m
1.50 m
1.62 m
0.85 m
1.82 m
1.250 s
1.229 s
0.841 s
1.34 m
1.72 m
1.85 m
1.46 m
1.60 m
0.82 m
1.86 m
1.251 s
1.228 s
0.842 s
1.37 m
1.62 m
1.89 m
1.50 m
1.63 m
0.83 m
1.81 m
1.153 s
1.081 s
1.081 s
1.35 m
1.71 m
1.86 m
1.49 m
1.62 m
0.85 m
1.82 m
1.316 s
0.981 s
0.894 s
(0.8)
H-26
H-27
H-28a
1.070 s
0.989 s
3.33 dd
(11.0; 1.1)
3.80 dd
(11.0; 1.7)
0.784 d
(6.8)
1.080 s
0.975 s
3.04 dd
(9.2; 0.9)
3.49 dd
(9.2; 1.7)
0.774 d
(6.7)
1.048 s
0.953 s
3.32 dd
(11.0; 1.2)
3.80 dd
(11.0; 1.8)
0.774 d
(6.8)
1.112 s
0.981 s
3.31 dd
(10.9; 1.2)
3.76 dd
(10.9; 1.7)
0.767 d
(6.8)
1.088 s
1.020 s
3.32 d
(10.9; 1.2)
3.76 dd
(10.9; 1.6)
0.776 d
(6.8)
H-28b
H-29
H-30
OMe
0.861 d
(7.2)
–
0.867 d
(6.9)
3.343 s
0.853 d
(6.8)
–
0.843 d
(6.8)
–
0.851 d
(6.9)
–
Collect. Czech. Chem. Commun. 2006, Vol. 71, No. 8, pp. 1131–1160

1154
Dračínský, Hybelbauerová, Sejbal, Buděšínský:
TABLE VI
1H NMR ch em ical sh ifts an d couplin g con stan ts of com poun ds 28, 30, 32 an d 33 in CDCl3
Proton
28
30
32
33
H-1α
H-1β
H-2α
H-2β
H-3α
0.92 m
1.69 m
1.65 m
1.65 m
1.53 m
1.92 m
1.80 m
1.98 dt
(14.0; 3.5)
1.39 m
2.10 ddd
(13.0; 6.2; 2.7)
2.22 ddd
(14.9; 4.5; 2.6)
2.77 td
(14.9; 14.9; 6.4)
–
2.77 ddd
(16.2; 11.3; 5.3)
2.36 ddd
(16.2; 9.8; 4.4)
–
1.75 m a
1.80 m a
3.14 m
(ΣJ = 15.8)
0.70 m
4.68 m
(ΣJ = 15.3)
–
3.43 t
(2.3)
H-5α
H-6
2.44 s
–
–
4.53 bs
3.12 bd
(4.1)
1.55m
H-7a
H-7b
1.63 m
1.67 m
2.02 d
(12.0)
2.4721 d
(12.0)
1.83 m
2.19 bd
(16.2)
1.83 m
H-9
1.35 m
1.35 m
1.52 m
1.20 m
1.43 m
1.78 m
1.02 m
1.77 m
1.20 m
1.83 m
1.37 m
1.78 m
1.88 m
1.46 m
1.61 m
0.87 m
1.73 m
1.056 s
1.156 s
1.210 s
1.372 s
0.927 s
3.84 dd
(11.1; 1.4)
4.26 dd
(11.1; 1.9)
0.768 d
(6.8)
1.94 m
1.43 m
1.50 m
1.30 m
1.68 m
1.68 m
0.89 m
1.73 m
1.19 m
1.95 m
1.36 m
1.73 m
1.86 m
1.50 m
1.60 m
0.85 m
1.84 m
1.101 s
1.489 s
1.136 s
1.106 s
1.067 s
3.32 dd
(10.8; 1.0)
3.73 dd
(10.8; 1.6)
0.785 d
(6.8)
1.85 m
1.39 m
1.60 m
1.21 m
1.61 m
1.55 m
1.04 m
1.74 m
1.16 m
1.94 m
1.33 m
1.68 m
1.85 m
1.45 m
1.62 m
0.83 m
1.80 m
0.879 s
1.261 s
1.014 s
1.075 s
0.970 s
3.31 dd
(11.0; 1.2)
3.75 dd
(11.0; 1.7)
0.769 d
(6.8)
1.35 m
1.34 m
1.42 m
1.14 m
1.65 m
1.70 m
1.05 m
1.75 m
1.27 m
1.82 m
1.40 m
1.78 m
1.88 m
1.45 m
1.62 m
0.88 m
1.74 m
0.746 s
1.101 s
1.101 s
1.191 s
0.923 s
3.87 bd
(11.1)
H-11a
H-11b
H-12a
H-12b
H-13
H-15a
H-15b
H-16a
H-16b
H-18
H-19
H-20
H-21a
H-21b
H-22a
H-22b
H-23
H-24
H-25
H-26
H-27
H-28a
H-28b
H-29
H-30
OAc
4.16 dd
(11.1; 1.8)
0.763 d
(6.8)
0.839 d
(6.8)
0.846 d
(6.8)
2.059 s
0.860 d
(6.9)
–
0.845 d
(6.8)
–
2.057 s
2.058 s
a
Sign als m ay be m utually in terch an ged.
Collect. Czech. Chem. Commun. 2006, Vol. 71, No. 8, pp. 1131–1160

B-Ring Substituted Lupane Derivatives
1155
TABLE VII
Vicin al couplin g con stan ts (in Hz) of rin g A proton s of 3-oxo derivatives
Couplin g con stan ts, Hz
Com pd
Con form ation
Substituen t
1α,2α
1α,2β
1β,2α
1β,2β
C
5.7
11.2
9.7
13.2
3.6
1.4
9.1
0
5.7
11.2
9.7
T1
T2
S
10.3
12.1
8.6
0.8
8.6
34a
12
15
16
22
26
30
32
7.9
5.9
9.4
12.7
10.2
10.4
3.6
4.4
3.0
1.6
1.8
6.6
2.6
2.6
5.3
7.8
7.0
8.7
8.7
9.8
6.2
6.4
9.8
24-oxo
5,6
∆
9.2
5,6
∆
9.3
6α-OH
12.0
4.6
6-oxo
14.8
14.9
4.4
6-oxo
4.5
5α,6α-epoxy
11.3
a
Couplin g con stan ts (in Hz) taken from ref.47
TABLE VIII
Results of m olecular m odellin g
Rin g A
con form ation
Num ber of
structures
En ergya
Θ1, °
Θ2, °
Subst.
kJ m ol–1
(C10–C1–C2–C3) (C1–C2–C3–C4)
24-oxo
C
53
21
23
2
341.7
345.1
346.7
377.5
–49
21
49
–57
–6
T1
sofa
oth er
–31
5,6
∆
T2
95
5
290.5
316.2
–31
–21
oth er
6α-OH
T1
88
7
339.0
345.9
347.5
24
–26
–51
–60
–14
47
sofa
C
5
6-oxo
C
59
37
3
339.0
346.3
352.6
–51
27
50
T1
–59
oth er
a
Lowest en ergy for a structure of given con form ation type.
Collect. Czech. Chem. Commun. 2006, Vol. 71, No. 8, pp. 1131–1160

1156
Dračínský, Hybelbauerová, Sejbal, Buděšínský:
3,6-Dioxolupan -28-yl Pivalate (26)
To a solution of diol 23 (97 m g, 0.18 m m ol) in pyridin e (5 m l), ch rom ium trioxide (145 m g,
1.45 m m ol) was added an d th e m ixture was stirred at room tem perature overn igh t. Th e m ix-
ture was extracted with eth er, th e eth er layer was wash ed with 2 M h ydroch loric acid, 5% so-
lution of potassium disulfite an d with saturated solution of sodium h ydrogen carbon ate an d
th en dried over an h ydrous sodium sulfate an d evaporated un der reduced pressure. Th e evap-
oration residue (74 m g, 77%) was pure en ough for use in an oth er reaction . A sam ple for
an alysis was obtain ed by m ultiple crystallisation from m eth an ol. M.p. 235–249 °C. EI-MS,
m/z (%): 540 (M+, 3), 524 (19), 509 (8), 425 (8), 409 (6), 217 (12), 203 (18), 191 (15), 57
(100). 1H NMR: 0.784 d, 3 H (H-29, J(29,30) = 6.8); 0.856 d, 3 H (H-30, J(30,20) = 6.8);
1.062 s, 3 H (H-27); 1.121 s, 3 H (H-26); 1.137 s, 3 H (H-25); 1.098 s, 1.488 s, 2 × 3 H (H-23
an d H-24); 1.206 s, 9 H (H pivalate); 2.02 d, 1 H (H-7a, J(7a,7b) = 11.9); 2.10 ddd, 1 H (H-1β,
J(1β,1α) = 13.1, J(1β,2β) = 6.0, J(1β,2α) = 2.6); 2.21 ddd, 1 H (H-2α, J(2α,2β) = 14.9,
J(2α,1α) = 4.6, J(2α,1β) = 2.6); 2.43 s, 1 H (H-5); 2.45 d, 1 H (H-7b, J(7b,7a) = 11.9); 2.77 td,
1 H (H-2β, J(2β,2α) = 14.8, J(2β,1α) = 14.8, J(2β,1β) = 6.2); 3.78 dd, 1 H (H-28a, J(28a,28b) =
11.2, Jl.r. = 1.3); 4.20 dd, 1 H (H-28b, J(28b,28a) = 11.2, Jl.r. = 1.9). For 13C NMR, see Table III.
Lupan e-3β,6β,28-triol (27)
To a solution of com poun d 26 (174 m g, 0.32 m m ol) in dry eth er (35 m l) lith ium alum in ium
h ydride (0.2 g, 5.2 m m ol) was added an d th e m ixture was refluxed for 3 h . Th en eth yl ace-
tate (25 m l, 315 m m ol) was added an d th e m ixture was refluxed for an oth er 30 m in . Th en
th e reaction m ixture was worked up in th e usual m an n er. Th e evaporation residue (148 m g)
was accordin g to TLC a m ixture of two com poun ds. Th e m ixture was separated by colum n
ch rom atograph y on silica gel (15 g) with eth er–h exan e (3:1) as eluen t. Followin g com -
poun ds were obtain ed an d crystallised from m eth an ol (in troduced accordin g to th eir in -
creasin g polarity).
3β,6β-Dihydroxylupan-28-yl acetate (28): 36 m g, 22%, m .p. 231–235 °C, [α]D –28. EI-MS,
m/z (%): 502 (M+, 4), 484 (12), 451 (10), 429 (35), 411 (11), 205 (60), 191 (55), 187 (100).
For 1H NMR, see Table VI. For 13C NMR, see Table III.
Lupane-3β,6β,28-triol (27): 60 m g, 41%, m .p. 241–244 °C, [α]D –28. IR: 3624 (OH), 1460,
1017. EI-MS, m/z (%): 460 (M+, 6), 442 (7), 429 (20), 411 (15), 393 (5), 259 (5), 235 (22), 207
(45), 205 (43), 187 (100), 177 (40), 123 (100). 1H NMR: 0.770 d, 3 H (H-29, J(29,20) = 6.8);
0.849 d, 3 H (H-30, J(30,20) = 6.8); 0.932 s, 3 H (H-27); 1.060 s, 3 H (H-23); 1.157 s, 3 H
(H-24); 1.209 s, 3 H (H-25); 1.354 s, 3 H (H-26); 3.14 m , 1 H (H-3, ΣJ = 15.7); 3.31 d, 1 H
(H-28a, J(28a,28b) = 11.0); 3.79 dd, 1 H (H-28b, J(28b,28a) = 11.0, Jl.r. = 1.4); 4.53 m , 1 H
(H-6α, ΣJ = 8.7). For C30H52O3 (460.7) calculated: 78.21% C, 11.38% H; foun d: 78.12% C,
11.48% H.
6β-Hydroxylupan e-3β,28-diyl Diacetate (29)
To th e solution of triol 27 (10 m g, 0.022 m m ol) in pyridin e (0.4 m l) acetic an h ydride
(0.4 m l, 4.4 m m ol) was added an d th e m ixture was left stan din g at room tem perature over-
n igh t. Th e m ixture was worked up in th e usual m an n er. Diacetate 29 (10 m g, 85%) was ob-
tain ed, m .p. 266–270 °C (ch loroform –m eth an ol), [α]D –21. IR: 3620 (OH), 1726 (acetate),
1461, 1387, 1367, 1252, 1031, 981. EI-MS, m/z (%): 544 (M+, 3), 526 (3), 466 (8), 451 (10),
411 (3), 205 (34), 187 (100), 123 (58). 1H NMR: 0.773 d, 3 H (H-29, J(29,30) = 6.7); 0.846 d,
Collect. Czech. Chem. Commun. 2006, Vol. 71, No. 8, pp. 1131–1160

B-Ring Substituted Lupane Derivatives
1157
3 H (H-30, J(30,20) = 6.9); 0.919 s, 3 H (H-27); 0.939 s, 3 H (H-23); 1.232 s, 3 H (H-24);
1.238 s, 3 H (H-25); 1.372 s, 3 H (H-26); 2.059 s, 6 H (2 × OAc); 3.83 dd, 1 H (H-28a,
J(28a,28b) = 11.0, Jl.r. = 1.4); 3.79 dd, 1 H (H-28b, J(28b,28a) = 11.0, Jl.r. = 1.8); 4.43 m , 1 H
(H-3α, ΣJ = 16.0); 4.52 bs, 1 H (H-6α). For 13C NMR, see Table III.
28-Hydroxylupan e-3,6-dion e (30)
To a solution of triol 27 (13 mg, 0.028 mmol) in pyridine (0.38 ml) bromine (20 µl, 0.40 mmol)
was added an d th e m ixture was left stan din g at –24 °C for 4 days. Th en th e m ixture was
diluted with water an d extracted with eth er. Th e eth er layer was wash ed with water, 2 M h y-
droch loric acid, 5% solution of sodium sulfite, with saturated solution of sodium h ydrogen -
carbon ate and with water and then it was evaporated under reduced pressure. Dione 30 (10 m g,
78%) was obtain ed, am orph ous, [α]D –49. IR: 3626 (OH), 1709 (C=O), 1459, 1396, 1386,
1367, 1026. EI-MS, m/z (%): 456 (M+, 25), 425 (100), 235 (15), 203 (15), 191 (40), 177 (25).
For 1H NMR, see Table VI. For 13C NMR, see Table III.
5α,6α-Epoxylupan e-3β,28-diol (31)
To a solution of olefin 18 (30 m g, 0.068 m m ol) in ch loroform (2 m l) 3-ch loroperoxyben zoic
acid (60%, 50 m g, 0.22 m m ol) was added an d th e m ixture was left stan din g at room tem per-
ature for 2.5 h . Th en eth er was added an d th e solution was wash ed with 5% solution of
potassium iodide an d th en with 5% solution of sodium sulfite to decolourise th e solution .
Th is procedure was repeated as lon g as th e potassium iodide solution ch an ged colour to
brown . Th en th e eth er layer was wash ed with water, dried over an h ydrous sodium sulfate
an d evaporated un der reduced pressure. Epoxide 31 was obtain ed (31 m g, 100%), m .p.
279 °C (ch loroform –eth an ol), [α]D –27. EI-MS, m/z (%): 458 (M+, 6), 440 (18), 235 (15), 275
(45), 203 (60), 175 (78), 135 (90), 121 (100). HRMS, m/z: 458.3742 (for C30H50O3 calculated:
458.3760). IR: 1463, 1388, 1238, 1064, 1021, 978, 952. 1H NMR: 0.761 d, 3 H (H-29,
J(29,20) = 6.9); 0.835 d, 3 H (H-30, J(30,20) = 7.0); 0.873 s, 3 H (CH3); 0.965 s, 3 H (H-27);
1.089 s, 6 H (2 × CH3); 1.120 s, 3 H (CH3); 2.28 dd, 1 H (H-7β, J(7β,7α) = 16.6, J(7β,6β) =
1.6); 3.15 dd, 1 H (H-6β, J(6β,7β) = 1.8, J(6β,7α) = 3.1); 3.30 dd, 1 H (H-28a, J(28a,28b) =
11.2, Jl.r. = 1.2); 3.54 m , 1 H (H-3α, ΣJ = 16.6); 3.74 dd, 1 H (H-28b, J(28b,28a) = 11.0, Jl.r.
1.8). For 13C NMR, see Table III.
=
5α,6α-Epoxy-28-h ydroxylupan -3-on e (32)
To th e solution of diol 31 (18 m g, 0.039 m m ol) in pyridin e (0.6 m l), brom in e (29 µl,
0.60 m m ol) was added an d th e m ixture was left stan din g at –24 °C for 4 days. Th en th e
m ixture was diluted with water an d extracted with eth er. Th e eth er layer was wash ed with
water, 2 M h ydroch loric acid, 5% solution of sodium sulfite, with saturated solution of so-
dium h ydrogen carbon ate an d with water. Th en it was dried over an h ydrous sodium sulfate
an d evaporated un der reduced pressure. Am orph ous title com poun d 32 (15 m g, 83%) was
obtain ed. EI-MS, m/z (%): 456 (M+, 20), 219 (80), 191 (100), 177 (70), 149 (100), 133 (88).
HRMS, m/z: 456.3590 (for C30H48O3 calculated: 456.3603). For 1H NMR, see Table VI. For
13C NMR, see Table III.
Collect. Czech. Chem. Commun. 2006, Vol. 71, No. 8, pp. 1131–1160

1158
Dračínský, Hybelbauerová, Sejbal, Buděšínský:
5β,6β-Epoxylupan e-3β,28-diyl Diacetate (33)
Olefin 19 (40 m g, 0.076 m m ol) was dissolved in a m ixture of dich lorom eth an e (0.8 m l),
eth an ol (1.2 m l) an d water (80 µl). Th e solution was treated with 70% aqueous perch loric
acid (60 µl) an d N-brom oacetam ide (40 m g) at room tem perature for 105 m in . Th e m ixture
was diluted with water, th e product extracted with eth er an d th e eth ereal solution was
wash ed with 5% aqueous sodium h ydrogen carbon ate solution , 5% aqueous sodium th io-
sulfate solution an d with water. Th en it was dried over an h ydrous sodium sulfate an d evap-
orated un der reduced pressure. Th e product was purified by preparative TLC, eth er–h exan e
(1:2) was used as eluen t. Epoxide 33 was obtain ed (16 m g, 39%), m .p. 257 °C (ch loro-
form –eth an ol), [α]D –20. IR: 1727, 1461, 1388, 1366, 1252, 1033, 983. EI-MS, m/z (%): 542
(M+, 10), 524 (3), 482 (13), 464 (6), 121 (100). HRMS, m/z: 542.3968 (for C34H54O5 calcu-
lated: 542.3971). For 1H NMR, see Table VI. For 13C NMR, see Table III.
The authors are indebted to Dr S. Hilgard for measurement of IR spectra. This work was
supported by the Grant Agency of Charles University (grants No. 429/2004/B-Ch/PrF and
No. 430/2004/B-Ch/PrF).
REFERENCES
1. Dračínský M., Richtr V., Křeček V., Sejbal J., Klinot J., Buděšínský M.: Collect. Czech.
Chem. Commun. 2006, 71, 387.
2. Connolly J. D., Hill R. A.: Methods Plant Biochem. 1991, 7, 331.
3. Kim D. S., Pezzuto J. M., Pisha E.: Bioorg. Med. Chem. Lett. 1998, 8, 1707.
4. Lee S. S., Chen W. C., Huang C. F., Su Y.: J. Nat. Prod. 1998, 61, 1343.
5. Fujikota T., Kashiwada Y., Kilkuskie R. E., Cosentino L. M., Ballas L. M., Jiang J. B., Chen
I.-S., Lee K.-H.: J. Nat. Prod. 1994, 57, 243.
6. Ali M., Ravinder E., Ramachandram R.: Pharmazie 2000, 55, 385.
7. Dantanarayana A. P., Kumar N. S., Sultanbawa M. U. S., Balasubramaniam S.: J. Chem.
Soc., Perkin Trans. 1 1981, 2717.
8. Narayanan C. R., Naik D. G.: Indian J. Chem., Sect., Sect. B: Org. Chem. Incl. Med. Chem.
1981, 20, 62.
9. Manzoor-I-Khuda M., Chowdhury S. A., Reza T., Chowdhury A. K.: J. Bangladesh Acad.
Sci. 1981, 5, 55.
10. Mustafa G., Anis E., Ahmed S., Anis I., Ahmed H., Malik A., Shahzad-ul-Hassan S.,
Choudhary M. I.: J. Nat. Prod. 2000, 63, 881.
11. Kuroyanaji M., Shiotsu M., Ebihara T., Kawai H., Ueno A., Fukushima L.: Chem. Pharm.
Bull. 1986, 34, 4012.
12. Lunardi I., Peixoto J. L. B., da Silva C. C., Shuqel I. T. A., Basso E. A., Vidotti G. J.:
J. Brazil. Chem. Soc. 2001, 12, 180.
13. Kouzi S. A., Chatterjee P., Pezzuto J. M., Hamann M. T.: J. Nat. Prod. 2000, 63, 1653.
14. Hess S. C., Delle Monache F.: J. Brazil. Chem. Soc. 1999, 10, 104.
15. Bilia A. R., Ciampi L., Mendez J., Morelli I.: Riv. Ital. EPPOS 1996, 7, 17.
16. Bilia A. R., Morelli I., Mendez J.: J. Nat. Prod. 1996, 59, 297.
17. Hisham A., Kumar G. J., Fujimoto Y., Hara N.: Phytochemistry 1996, 42, 789.
Collect. Czech. Chem. Commun. 2006, Vol. 71, No. 8, pp. 1131–1160

B-Ring Substituted Lupane Derivatives
1159
18. Facundo V. A., Andrade C. H. S., Silveria E. R., Braz-Filho R., Hufford C. D.:
Phytochemistry 1993, 32, 411.
19. Drewes S. E., Mashimbye M. J., Field J. S., Ramesar N.: Phytochemistry 1991, 30, 3490.
20. Dantanarayana A. P., Kumar N. S., Muthukuda P. M., Wazeer M. I. M.: Phytochemistry
1982, 21, 2065.
21. Dantanarayana A. P., Kumar N. S., Muthukuda P. M., Balasubramaniam S.: Phytochemistry
1983, 22, 473.
22. Adnyana I. K., Tezuka Y., Awale S., Banskota A. H., Tran K. Q., Kadota S.: Chem. Pharm.
Bull. 2000, 48, 1114.
23. Adnyana I. K., Tezuka Y., Banskota A. H., Xiong Q., Tran K. Q., Kadota S.: J. Nat. Prod.
2000, 63, 496.
24. Adnyana I. K., Tezuka Y., Banskota A. H., Tran K. Q., Kadota S.: J. Nat. Prod. 2001, 64,
360.
25. Jiang Z.-H., Tanaka T., Kouno I.: Phytochemistry 1995, 40, 1223.
26. Anaya J., Caballero M. C., Grande M., Navarro J. J., Tapia I., Almeida J. F.: Phytochemistry
1989, 28, 2206.
27. Rasool N., Khan A. Q., Malik A.: J. Nat. Prod. 1989, 52, 749.
28. Fukunaya T., Nishiya K., Kajikawa I., Watanabe Y., Suzuki N., Takeya K., Itokawa H.:
Chem. Pharm. Bull. 1988, 36, 1180.
29. Chatterjee P., Kouzi S. A., Pezzuto J. M., Hamann M. T.: Appl. Environ. Microbiol. 2000,
66, 3850.
30. Akihisa T., Kimura Y., Kasahara Y., Kumaki K., Thakur S., Tamura T.: Phytochemistry
1997, 46, 1261.
31. Huang H.-C., Shen C.-C., Chen C.-F., Wu Y.-C., Kuo Y.-H.: Chem. Pharm. Bull. 2000, 48,
1079.
32. Atta-ur-Rahman, Khan M. A., Khan N. H.: Phytochemistry 1973, 12, 3004.
33. Rodriguez-Gamboa T., Fernandes J. B., Rodrigues E., da Silva M. F. D. F., Vieira P. C.,
Barrios M., Castro-Castillo O., Victor S. R., Pagnocca F. C., Bueno O. C., Hebling M. J. A.:
J. Brazil. Chem. Soc. 2001, 12, 386.
34. Schuhly W., Heilmann J., Calis I., Sticher O.: Planta Med. 1999, 65, 740.
35. Talapatra S. K., Shrestha K. M., Pal M. K., Basak A., Talapatra B.: Phytochemistry 1989, 28,
3437.
36. Siddiqui S., Begum S., Hafeez F., Siddiqui B. S.: Phytochemistry 1989, 28, 1187.
37. Adnyana I. K., Tezuka Y., Banskota A. H., Tran K. Q., Kadota S.: Biol. Pharm. Bull. 2000,
23, 1328.
38. Endová M., Klinotová E., Sejbal J., Máca B., Klinot J., Protiva J.: Collect. Czech. Chem.
Commun. 1994, 59, 1420.
39. Klinotová E., Křeček V., Klinot J., Endová M., Eisenreichová J., Buděšínský M., Štícha M.:
Collect. Czech. Chem. Commun. 1997, 62, 1776.
40. Cohen K. F., Kazlauskas R., Pinhey J. T.: J. Chem. Soc., Perkin Trans. 1 1973, 2076.
41. Klinot J., Hořejší M., Buděšínský M., Vystrčil A.: Collect. Czech. Chem. Commun. 1975,
40, 3712.
42. Kirk D. N., Hartshorn M. P.: Steroid Reaction Mechanisms, p. 168. Elsevier, Amsterdam
1968.
43. Midgley J. M., Millard B. J., Whalley W. B.: J. Chem. Soc. 1971, 13.
44. King F. E., King T. J.: J. Chem. Soc. 1956, 4469.
45. Pouzar V., Slavíková T., Černý I.: Collect. Czech. Chem. Commun. 1997, 62, 109.
Collect. Czech. Chem. Commun. 2006, Vol. 71, No. 8, pp. 1131–1160

1160
Dračínský, Hybelbauerová, Sejbal, Buděšínský:
46. Kočovský P., Černý V.: Collect. Czech. Chem. Commun. 1979, 44, 2275.
47. Klinot J., Buděšínský M., Klinotová J., Vystrčil A.: Collect. Czech. Chem. Commun. 1986,
51, 593.
48. Tsuda Y., Kiuchi F.: Chem. Pharm. Bull. 1984, 32, 4806.
49. Klinot J., Podlaha J., Podlahová J., Malý K., Petříček V.: Collect. Czech. Chem. Commun.
1989, 54, 725.
50. Haasnoot C. A. G., de Leeuw F. A. A. M., Altona C.: Tetrahedron 1980, 3267.
51. Ruzicka L., Isler O.: Helv. Chim. Acta 1936, 19, 506.
Collect. Czech. Chem. Commun. 2006, Vol. 71, No. 8, pp. 1131–1160