VOL.
XXXII
NO.
3
THE
JOURNAL
OF
ANTIBIOTICS
THE SYNTHESIS OF e-RHODOMYCINONE- AND CARMINOMYCIN11-METHYL ETHERS
(excess
Sir: In recent years, the anthracycline antibiotics adriamycin (la) and daunomycin (1b) have been shown to be clinically effective anti-tumor agents. Carminomycin (1c), isolated from Actinomadura carminata in the Soviet Union,1) has a similar antitumor spectrum to adriamycin but with less cardiotoxic potential.2) Since daunomycin of
is
the
4-methyl
carminomycin, in
and/or
11-methyl
mine
ether
in
in
Direct
to
to
products,
give
on
fact
and
potassium
bonate
gave
a
which A
183°C.
C,
That
comparisons
7.5
Ia.
R =CH3
phosphate
Ib.
R = R'=
one
of
the
aromatic Since the
Ic.
R = H. R'=
R = Ac
3b.
R = H
3c.
R = CH3
5a.
R=R'=
H
Sb.
R=R'-Ac
Sc.
R =Ac.
4b.R=
the
Methylation
64
only
acetates aromatic spectrum
product a
C4
was acetate
product, was
4,6,7-triacetate isomeric
and
4, 7,11
mp
either (3b)
of
the UV
anthraquinones.
4a.
mixof
the
for 1),
not
IR
isomeric spectral The
R=H
2b.
R =CH3
the or
-triacetate. this
phenol
the
R'=
H
R =H
e-
4.84%.
2o.
CH3
acetone-
pmr
hydrolysis
characteristic
245-247•Ž,
other
and by
CH3
and
buffer
hydrolysis
(Table
shown
, R' = CH2OH
3a. the
after an
3),
the
with
product,
181 Calcd.
H,
same
to
that in
(4:
of
was
mp
4.84%.
61.19;
an-
(3a),
ture
region
the
H, C,
Attention
22•Ž
occurred.
61.42;
gave
(2b),
pattern
directed
found at
was ether,
ether
e-
an the
7,11-tetraacetate3) was
this
6-methyl
e-rhodomycinone-4,6,-
hours
Found: C23H2209•0.5H2O:
22•Ž)
diazo-
with
therefore
pH
I-methyl
(3c
remaining bicarbonate-
at
to
with
carminomycin.
it
rhodomycinone-1
for
sodium
4 hours
result
oxidation
known
aqueous
stirred
mix-
(2a),
aromatic
(5y. 1
the
car-
reacted
thracyclinone
lc
difficult
rhodomycinone
of
of
when
was
1:
ether
of
N-
methyl
similar
obtained
methane
groups
acetone
mono-methyl
of
complex
was
separate.
was
the was
the
with
iodide
was
of 1e
derivative
methylation
ture
a Removal
a
reflux
car-
a mixture in
trifluoroacetyl
afforded isomer).
with
under
series.
of
and
C6
acetone
deter-
methylation
expected
hours)
its
dry
carbonate
rela-
this
functions
6.5
or
in
potassium
6-
of
order
phenolic
for
acetyl
iodide
of
in-
the
structure-activity
tionships
as
were
preparing
minomycin
methyl
excess
ether
we
terested
of
10%
247
in-
248
THE
Table
JOURNAL
1. 1H-NMR
OF
chemical
ANTIBIOTICS
shifts of selected
MAR.
1979
Phenolic
OH
protons.*
Position Compound
1
3
10
7
OCH3
13.05(s) 12.33(s) 111 .48(s)
2a
7.76
7.54(m)
7.24(dd)
5.32(s)
4.26(s)
3a
7.71(t) 7. 74(t) 7.72(t)
7.36(dd) 7.38(dd) 7.35(dd)
6.48(b)
3c
8.07(dd) 8.21(dd) 8.10(dd)
6.44(b)
4.22(b) 4.37(s) 4.37(s)
3.93(s)
2b
7.80
7.53(m)
7.I8(dd)
5.33(b)
4.20(s)
3.80(s)
5a
7.83
7.60(m)
7.24(dd)
5.16(s)
3.07(A B)
5b
8.12(dd) 8.20(dd) 7.90
7.76(t) 7.74(t) 7.60(m)
7.41(dd) 7.38(dd) 7.26(dd)
5.20(b)
3.12(b) 3.17(AB) 3.21(A B)
3b
5c 6
*
All
spectra
were
recorded
standard.
s=singlet,
qua-tet,
signal, m=multiplet.
Table
No. 1
2b (in CDCl3)
119.5
119.7 137.2 123.5 161.7 115.0 191.9 113.9 153.4 134.3 62.4 34.0 71.4
3
125.0
4
162.1
4a
115.8
5
190.1
5a
110.9
6
156.6
6a
134.7
7
61.2
8
35.0
9 *
71.5 In ppm downfield
pounds
2. 13C-NMR
2a (in DMSO-d6)
137.7
such as those
from
tetramethylsilane;
described
5.30(b)
in CDC1
as
Carbon
5.16(b)
100 MHz
tetramethylsilane b=broad
internal
at
6.38(b)
13.50(s)
13.20(s) 3.91(s)
3 and chemical shifts are t -triplet, dd=doublet
chemical
13.06(s) 11 .73(s) (13.03(s) 12.48(s) '11 .60(b)
not observed
reported in ppm from of doublets, AB= AB
shifts.*
Carbon
No.
2a (in DMSO-De)
10 10a 11
2b (in CDCI•.)
51.8
52.3
132.9
135.0
156.7
157.8
111.0
122.0
12
185.9
180.0
12a
139.3
140.3
13
32.7
32.5
14
7.1
6.8
15
171.3
171.2
16
52.6
11 a
52.2
OCH3
assignments
were
61.3
based
on comparison
with known
cony
by DOYLE et al.7)
frared spectra of 2b (both as KBr disc and in chloroform solution) showed absorptions for both non-bonded (1670 cm-1) and hydrogenbonded (1620 cm-l) quinone carbonyl groups which is typical of a 1,8-dihydroxyanthraquinone and not that expected for a 1,5-dihydroxyanthra-
that
quinone (which would show only a single carbonyl absorption).4) Similarly, aklavinone (4a) gives absorption bands at 1674 and 1623 cm-1,5) while e-rhodomycinone (2a) shows only bonded
180.0 ppm (Table 2) indicating that one of these carbonyls is bis-hydrogen bonded (the downfield one) and the other is non-bonded.6) A similar sequence of reactions was then carried out on N-trifluoroacetyl carminomycin (5a, prepared from 1c and trifluoroacetic anhyd-
quinone absorption at 1615 cm-1. The UVvisible spectrum of 2b was also very similar to
of the 1,8-dihydroxyanthraquinone
aklavin
(4b), showing a single absorption maximum in the visible region at 448 nm which moved to 525 nm on addition of sodium hydroxide. Finally, the quinone carbonyl resonances in the 13C nmr spectrum of 2b were at 191 .9 and
VOL. XXXII
ride
in
NO.
3
dichloromethane
at
chromatography
over
acetate).
by
addition
tetraacetate
to
(5b)
214-266•Ž.
as
Calcd.
4.56;
N,
an
1.76%.
Hydrolysis
at
sodium 22•Ž
traction triacetate 247•Ž. Calcd.
by
(5c)
as :
of
H, • H2O: of
orange H,
C34H32F3NO14:
5
for
1.5
and
ex-
the of
245
N,
55.59;
The authors wish to thank Dr. W. T. BRADNER of these laboratories for the in vivo anti-tumor data. JOHN M. ESSERY* TERRENCE W. DOYLE
4,6,4'-
mp
4.39;
C,
Acknowledgement
54.34;
(acetone-
3:
4,6,7,11-tetraacetate is described. This series of reactions was used as a model for the conversion of carminomycin to its 11-methyl ether. The anti-tumor activity of the latter compound was less than that of both carminomycin and its 4-methyl ether (daunomycin).
N,
C,
5b
solid
55.51;
mp
4.54;
-
Research Division, Bristol Laboratories, P.O. Box 657, Syracuse, New York 13201, U.S.A.
1.90%.
H,
of
5c
4.45;
N,
iodide
added
at
0.5,
3,
reflux
period
of
16
hours
a
final
addition)
gave
purified.
It
0 .1
and
B
buffered This
to
pH
(from
ether).
2.39%. H,
tion
N,
spectrum
and
4a,
bonded The
and
those
and
ether
L-1210
leukenmic
value
of
136
(lc,
used
as
0.8
mg/kg.
1c
and
16
the
6
both
of
a further
phase. 186•Ž 5.93;
N,
those
both
6
was
confirming
vitro
of
groups.
that
had
a
whereas had
similar it
tested
the
maximum
T/C
carminomycin a
in
was against
value
of
comparison and
2b
hydrogen-
very
When 6
absorp-
carbonyl
of
mg/kg
C,
infrared
to
detailed in
C18/
acetate
H,
to
due
control)
Amore
on
185.
60.26;
shown. mice,
at
22•Ž)
liquid
mp
quinone
4b,
as
the of
vivo
157
at
between will
be
the
communication.
Summary The methyl
conversion ether
of e-rhodomycinone via
selective
to hydrolysis
References
not
1)
at
HPLC
similar
spectrum 2b
l hour
The
was
nonbonded
of
11-methyl
subject
6
(Received December 20, 1978) the
C27H29NO1O.0.5H2O:
maxima
UV-visible
as
2.61%;,.
of
showing
of
hours,
was
M sodium
solid C,
for
5.64;
which
by
45)
red
Found:
Calcd.
60.44;
(55:
10
following
-0.1
a
and
5 for
purified
4.0 6 as
carexcess
(tetrahydrofuran 2:
methanol
provided
an 6
hydrolysed
was
using
with
ether
hydroxide
product
Porasil
reflux
a methyl
was
N sodium the
at
potassium
methyl with
acetone
(anhydrous
in
249
ANTIBIOTICS
.
Methylation
to
22•Ž
4,6,11,4'-
solid
bonate
-
the
provided
an C,
pyri-
at
acidification
chloroform)
Found for
1.99%
gave
bicarbonate,
followed
into
of
hours
54.58;
C36H34F3NO15
ethyl
excess 2.5
OF
by
using
off-white C,
for
aqueous
hours
an
1 for water)
Found:
1.91%
5%
1:
JOURNAL
followed
gel
(with
anhydride
followed
22•Ž
silica
Acetylation
dine-acetic
H,
THE
its of
11the
GAUZE,G. F.: M. A. SVESHNIKOVA, R. S. UKHOLINA, G. V. GAVRILINA, V. A. FILICHEVA & E. G. GLADKIKH: Production of antitumor antibiotic carminomycin by Actinomadur•a carminata sp. nov. Antibiotiki 18: 675-678, 1973 2) CROOKE, S. T.: A review of carminomycin, a new anthracycline developed in the USSR. J. Medicine 8: 295 - 316, 1977 3) BROCKMANN, H. & H. BROCKMANN, Jr.: e-Rhodomycinon. Chem. Ber. 94: 2681-2694, 1961 4) BLOOM, H.; L. H. BRIGGS & B. CLEVERLEY: Physical properties of anthraquinones and its derivatives. 1. Infrared spectra. J. Chem. Soc. 1959: 178.-185, 1959 5) GORDON, J. J.; L. M. JACKMAN,W. D. OLLIS & 1. 0. SUTHERLAND: Aklavinone. Tetrahed. Lett. 1960: 28 - 34, 1960 6) Cox, R. H.; F. CHURCHILL,R. J. COLE& J. W. DORNER: Carbon-13 nuclear magnetic resonance studies of the structure and biosynthesis of versiconal acetate. J. Am. Chem. Soc. 99: 3159 - 3161, 1977 7) DOYLE, T. W.: D. E. NETTLETON,R. E. GRULICH, D. M. BALIT7, D. L. JOHNSON& A. L. VULCANO: Antitumor agents from the bohemic acid complex. (submitted to J. Am. Chem. Soc.)
XXXII
NO.
3
THE
JOURNAL
OF
ANTIBIOTICS
THE SYNTHESIS OF e-RHODOMYCINONE- AND CARMINOMYCIN11-METHYL ETHERS
(excess
Sir: In recent years, the anthracycline antibiotics adriamycin (la) and daunomycin (1b) have been shown to be clinically effective anti-tumor agents. Carminomycin (1c), isolated from Actinomadura carminata in the Soviet Union,1) has a similar antitumor spectrum to adriamycin but with less cardiotoxic potential.2) Since daunomycin of
is
the
4-methyl
carminomycin, in
and/or
11-methyl
mine
ether
in
in
Direct
to
to
products,
give
on
fact
and
potassium
bonate
gave
a
which A
183°C.
C,
That
comparisons
7.5
Ia.
R =CH3
phosphate
Ib.
R = R'=
one
of
the
aromatic Since the
Ic.
R = H. R'=
R = Ac
3b.
R = H
3c.
R = CH3
5a.
R=R'=
H
Sb.
R=R'-Ac
Sc.
R =Ac.
4b.R=
the
Methylation
64
only
acetates aromatic spectrum
product a
C4
was acetate
product, was
4,6,7-triacetate isomeric
and
4, 7,11
mp
either (3b)
of
the UV
anthraquinones.
4a.
mixof
the
for 1),
not
IR
isomeric spectral The
R=H
2b.
R =CH3
the or
-triacetate. this
phenol
the
R'=
H
R =H
e-
4.84%.
2o.
CH3
acetone-
pmr
hydrolysis
characteristic
245-247•Ž,
other
and by
CH3
and
buffer
hydrolysis
(Table
shown
, R' = CH2OH
3a. the
after an
3),
the
with
product,
181 Calcd.
H,
same
to
that in
(4:
of
was
mp
4.84%.
61.19;
an-
(3a),
ture
region
the
H, C,
Attention
22•Ž
occurred.
61.42;
gave
(2b),
pattern
directed
found at
was ether,
ether
e-
an the
7,11-tetraacetate3) was
this
6-methyl
e-rhodomycinone-4,6,-
hours
Found: C23H2209•0.5H2O:
22•Ž)
diazo-
with
therefore
pH
I-methyl
(3c
remaining bicarbonate-
at
to
with
carminomycin.
it
rhodomycinone-1
for
sodium
4 hours
result
oxidation
known
aqueous
stirred
mix-
(2a),
aromatic
(5y. 1
the
car-
reacted
thracyclinone
lc
difficult
rhodomycinone
of
of
when
was
1:
ether
of
N-
methyl
similar
obtained
methane
groups
acetone
mono-methyl
of
complex
was
separate.
was
the was
the
with
iodide
was
of 1e
derivative
methylation
ture
a Removal
a
reflux
car-
a mixture in
trifluoroacetyl
afforded isomer).
with
under
series.
of
and
C6
acetone
deter-
methylation
expected
hours)
its
dry
carbonate
rela-
this
functions
6.5
or
in
potassium
6-
of
order
phenolic
for
acetyl
iodide
of
in-
the
structure-activity
tionships
as
were
preparing
minomycin
methyl
excess
ether
we
terested
of
10%
247
in-
248
THE
Table
JOURNAL
1. 1H-NMR
OF
chemical
ANTIBIOTICS
shifts of selected
MAR.
1979
Phenolic
OH
protons.*
Position Compound
1
3
10
7
OCH3
13.05(s) 12.33(s) 111 .48(s)
2a
7.76
7.54(m)
7.24(dd)
5.32(s)
4.26(s)
3a
7.71(t) 7. 74(t) 7.72(t)
7.36(dd) 7.38(dd) 7.35(dd)
6.48(b)
3c
8.07(dd) 8.21(dd) 8.10(dd)
6.44(b)
4.22(b) 4.37(s) 4.37(s)
3.93(s)
2b
7.80
7.53(m)
7.I8(dd)
5.33(b)
4.20(s)
3.80(s)
5a
7.83
7.60(m)
7.24(dd)
5.16(s)
3.07(A B)
5b
8.12(dd) 8.20(dd) 7.90
7.76(t) 7.74(t) 7.60(m)
7.41(dd) 7.38(dd) 7.26(dd)
5.20(b)
3.12(b) 3.17(AB) 3.21(A B)
3b
5c 6
*
All
spectra
were
recorded
standard.
s=singlet,
qua-tet,
signal, m=multiplet.
Table
No. 1
2b (in CDCl3)
119.5
119.7 137.2 123.5 161.7 115.0 191.9 113.9 153.4 134.3 62.4 34.0 71.4
3
125.0
4
162.1
4a
115.8
5
190.1
5a
110.9
6
156.6
6a
134.7
7
61.2
8
35.0
9 *
71.5 In ppm downfield
pounds
2. 13C-NMR
2a (in DMSO-d6)
137.7
such as those
from
tetramethylsilane;
described
5.30(b)
in CDC1
as
Carbon
5.16(b)
100 MHz
tetramethylsilane b=broad
internal
at
6.38(b)
13.50(s)
13.20(s) 3.91(s)
3 and chemical shifts are t -triplet, dd=doublet
chemical
13.06(s) 11 .73(s) (13.03(s) 12.48(s) '11 .60(b)
not observed
reported in ppm from of doublets, AB= AB
shifts.*
Carbon
No.
2a (in DMSO-De)
10 10a 11
2b (in CDCI•.)
51.8
52.3
132.9
135.0
156.7
157.8
111.0
122.0
12
185.9
180.0
12a
139.3
140.3
13
32.7
32.5
14
7.1
6.8
15
171.3
171.2
16
52.6
11 a
52.2
OCH3
assignments
were
61.3
based
on comparison
with known
cony
by DOYLE et al.7)
frared spectra of 2b (both as KBr disc and in chloroform solution) showed absorptions for both non-bonded (1670 cm-1) and hydrogenbonded (1620 cm-l) quinone carbonyl groups which is typical of a 1,8-dihydroxyanthraquinone and not that expected for a 1,5-dihydroxyanthra-
that
quinone (which would show only a single carbonyl absorption).4) Similarly, aklavinone (4a) gives absorption bands at 1674 and 1623 cm-1,5) while e-rhodomycinone (2a) shows only bonded
180.0 ppm (Table 2) indicating that one of these carbonyls is bis-hydrogen bonded (the downfield one) and the other is non-bonded.6) A similar sequence of reactions was then carried out on N-trifluoroacetyl carminomycin (5a, prepared from 1c and trifluoroacetic anhyd-
quinone absorption at 1615 cm-1. The UVvisible spectrum of 2b was also very similar to
of the 1,8-dihydroxyanthraquinone
aklavin
(4b), showing a single absorption maximum in the visible region at 448 nm which moved to 525 nm on addition of sodium hydroxide. Finally, the quinone carbonyl resonances in the 13C nmr spectrum of 2b were at 191 .9 and
VOL. XXXII
ride
in
NO.
3
dichloromethane
at
chromatography
over
acetate).
by
addition
tetraacetate
to
(5b)
214-266•Ž.
as
Calcd.
4.56;
N,
an
1.76%.
Hydrolysis
at
sodium 22•Ž
traction triacetate 247•Ž. Calcd.
by
(5c)
as :
of
H, • H2O: of
orange H,
C34H32F3NO14:
5
for
1.5
and
ex-
the of
245
N,
55.59;
The authors wish to thank Dr. W. T. BRADNER of these laboratories for the in vivo anti-tumor data. JOHN M. ESSERY* TERRENCE W. DOYLE
4,6,4'-
mp
4.39;
C,
Acknowledgement
54.34;
(acetone-
3:
4,6,7,11-tetraacetate is described. This series of reactions was used as a model for the conversion of carminomycin to its 11-methyl ether. The anti-tumor activity of the latter compound was less than that of both carminomycin and its 4-methyl ether (daunomycin).
N,
C,
5b
solid
55.51;
mp
4.54;
-
Research Division, Bristol Laboratories, P.O. Box 657, Syracuse, New York 13201, U.S.A.
1.90%.
H,
of
5c
4.45;
N,
iodide
added
at
0.5,
3,
reflux
period
of
16
hours
a
final
addition)
gave
purified.
It
0 .1
and
B
buffered This
to
pH
(from
ether).
2.39%. H,
tion
N,
spectrum
and
4a,
bonded The
and
those
and
ether
L-1210
leukenmic
value
of
136
(lc,
used
as
0.8
mg/kg.
1c
and
16
the
6
both
of
a further
phase. 186•Ž 5.93;
N,
those
both
6
was
confirming
vitro
of
groups.
that
had
a
whereas had
similar it
tested
the
maximum
T/C
carminomycin a
in
was against
value
of
comparison and
2b
hydrogen-
very
When 6
absorp-
carbonyl
of
mg/kg
C,
infrared
to
detailed in
C18/
acetate
H,
to
due
control)
Amore
on
185.
60.26;
shown. mice,
at
22•Ž)
liquid
mp
quinone
4b,
as
the of
vivo
157
at
between will
be
the
communication.
Summary The methyl
conversion ether
of e-rhodomycinone via
selective
to hydrolysis
References
not
1)
at
HPLC
similar
spectrum 2b
l hour
The
was
nonbonded
of
11-methyl
subject
6
(Received December 20, 1978) the
C27H29NO1O.0.5H2O:
maxima
UV-visible
as
2.61%;,.
of
showing
of
hours,
was
M sodium
solid C,
for
5.64;
which
by
45)
red
Found:
Calcd.
60.44;
(55:
10
following
-0.1
a
and
5 for
purified
4.0 6 as
carexcess
(tetrahydrofuran 2:
methanol
provided
an 6
hydrolysed
was
using
with
ether
hydroxide
product
Porasil
reflux
a methyl
was
N sodium the
at
potassium
methyl with
acetone
(anhydrous
in
249
ANTIBIOTICS
.
Methylation
to
22•Ž
4,6,11,4'-
solid
bonate
-
the
provided
an C,
pyri-
at
acidification
chloroform)
Found for
1.99%
gave
bicarbonate,
followed
into
of
hours
54.58;
C36H34F3NO15
ethyl
excess 2.5
OF
by
using
off-white C,
for
aqueous
hours
an
1 for water)
Found:
1.91%
5%
1:
JOURNAL
followed
gel
(with
anhydride
followed
22•Ž
silica
Acetylation
dine-acetic
H,
THE
its of
11the
GAUZE,G. F.: M. A. SVESHNIKOVA, R. S. UKHOLINA, G. V. GAVRILINA, V. A. FILICHEVA & E. G. GLADKIKH: Production of antitumor antibiotic carminomycin by Actinomadur•a carminata sp. nov. Antibiotiki 18: 675-678, 1973 2) CROOKE, S. T.: A review of carminomycin, a new anthracycline developed in the USSR. J. Medicine 8: 295 - 316, 1977 3) BROCKMANN, H. & H. BROCKMANN, Jr.: e-Rhodomycinon. Chem. Ber. 94: 2681-2694, 1961 4) BLOOM, H.; L. H. BRIGGS & B. CLEVERLEY: Physical properties of anthraquinones and its derivatives. 1. Infrared spectra. J. Chem. Soc. 1959: 178.-185, 1959 5) GORDON, J. J.; L. M. JACKMAN,W. D. OLLIS & 1. 0. SUTHERLAND: Aklavinone. Tetrahed. Lett. 1960: 28 - 34, 1960 6) Cox, R. H.; F. CHURCHILL,R. J. COLE& J. W. DORNER: Carbon-13 nuclear magnetic resonance studies of the structure and biosynthesis of versiconal acetate. J. Am. Chem. Soc. 99: 3159 - 3161, 1977 7) DOYLE, T. W.: D. E. NETTLETON,R. E. GRULICH, D. M. BALIT7, D. L. JOHNSON& A. L. VULCANO: Antitumor agents from the bohemic acid complex. (submitted to J. Am. Chem. Soc.)
