US 5,108,912 AGranted

Antitumor antibiotics (LL-E33288 complex)

AssigneeAmerican Cyanamid Company

InventorsMay D. Lee, Michael Greenstein, David P. Labeda, Amedeo A. Fantini

Life of this patent

Estimating the term…

Abstract

Antibacterial and antitumor agents designated LL-E33288 complex and their production by new strains of Micromonospora echinospora spp. calichensis NRRL-15839, NRRL-15975 and NRRL-18149, are disclosed.

Claims7

Dependent claims

A process for producing antibiotics LL-E33288.alpha..sub.1 -Br having the following R.sub.f values in the indicated solvent systems on TLC on silica gel sheets:

a) ethyl acetate saturated with 0.1M aqueous potassium dihydrogen phosphate, R.sub.f =0.67;
b) 3% isopropanol in ethyl acetate saturated with 0.1M aqueous potassium dihydrogen phosphate, R.sub.f =0.80;
and
c) ethyl acetate:
methanol (95:5), R.sub.f =0.79;
LL-E33288.alpha..sub.2 -Br having the following R.sub.f values in the indicated solvent systems on TLC on silica gel sheets:
a) ethyl acetate saturated with 0.1M aqueous potassium dihydrogen phosphate, R.sub.f =0.61;
b) 3% isopropanol in ethyl acetate saturated with 0.1M aqueous potassium dihydrogen phosphate, R.sub.f =0.75;
and
c) ethyl acetate:
methanol (95:5), R.sub.f =0.73;
and having the following structure:
##STR5## LL-E33288.alpha..sub.3 -Br having the following R.sub.f values in the indicated solvent systems on TLC on silica gel sheets:
a) ethyl acetate saturated with 0.1M aqueous potassium dihydrogen phosphate, R.sub.f =0.55;
b) 3% isopropanol in ethyl acetate saturated with 0.1M aqueous potassium dihydrogen phosphate, R.sub.f =0.69;
and
c) ethyl acetate:
methanol (95:5), R.sub.f =0.61;
and having the following structure:
##STR6## LL-E33288.alpha..sub.4 -Br having the following R.sub.f values in the indicated solvent systems on TLC on silica gel sheets:
a) ethyl acetate saturated with 0.1M aqueous potassium dihydrogen phosphate, R.sub.f =0.49;
b) 3% isopropanol in ethyl acetate saturated with 0.1M aqueous potassium dihydrogen phosphate, R.sub.f =0.64;
and
c) ethyl acetate:
methanol (95:5), R.sub.f =0.54;
LL-E33288.beta..sub.1 -Br
a) having an approximate elemental analysis:
C 48.6;
H 5.6;
N 2.9;
S 9.1 and Br 5.5;
b) having a melting point:
146.degree.-150.degree. C. (dec.);
c) having a specific rotation:
[.alpha.]D.sup.26 =-49.degree..+-.10.degree. (0.1%, ethanol);
d) having ultraviolet absorption spectra as shown in FIG. I of the drawings;
e) having an infrared absorption spectrum as shown in FIG. II of the drawings;
f) having a proton magnetic resonance spectrum as shown in FIG. III of the drawings;
g) having a carbon-13 magnetic resonance spectrum as shown in FIG. IV of the drawings with significant peaks at:
______________________________________ 17.60(q) 17.64(q) 18.9(q) 19.7(q) 22.4(q) 22.8(q) 23.5(q) 34.3(t) 36.9(t) 39.2(t/d) 47.8(d) 51.7(q) 52.7(q) 54.6(d) 56.3(q) 57.2(q) 57.8(d) 61.0(q/d) 61.7(d) 62.4(t) 66.9(d) 68.4(d) 69.1(d) 69.7(d) 70.2(d) 71.1(d) 71.9(d) 72.1(s) 76.1(d) 81.0(d) 83.3(s) 88.2(s) 97.4(d) 99.7(d) 100.8(s) 102.5(d) 115.1(s) 123.4(d) 124.4(d) 126.5(d) 130.2(s) 130.8(s) 144.6(s) 149.3(s) 149.5(s) 191.7(s) 192.4(s) and ______________________________________
h) having the following R.sub.f values in the indicated solvent systems on TLC on silica gel sheets:
(i) ethyl acetate saturated with 0.1M potassium dihydrogen phosphate, R.sub.f =0.24;
(ii) 3% isopropanol in ethyl acetate saturated with 0.1M aqueous potassium dihydrogen phosphate, R.sub.f =0.35;
(iii) ethyl acetate:
methanol (95:5), R.sub.f =0.36.
i) having a molecular weight:
1333/1335, respectively for .sup.79 Br/.sup.81 Br;
j) having a molecular formula:
C.sub.56 H.sub.76 N.sub.3 O.sub.21 S.sub.4 Br;
k) and having the structure ##STR7## LL-E33288.beta..sub.2 -Br having the following R.sub.f values in the indicated solvent systems on TLC on silica gel sheets:
a) ethyl acetate saturated with 0.1M aqueous potassium dihydrogen phosphate, R.sub.f =0.32;
b) 3% isopropanol in ethyl acetate saturated with 0.1M aqueous potassium dihydrogen phosphate, R.sub.f =0.41;
and
c) ethyl acetate:
methanol (95:5), R.sub.f =0.45;
and LL-E33288.gamma..sub.1 -Br
a) having ultraviolet absorption spectra as shown in FIG. V of the drawings;
b) having an infrared absorption spectrum as shown in FIG. VI of the drawings;
c) having a proton magnetic resonance spectrum as shown in FIG. VII of the drawings;
and
d) having the following R.sub.f values in the indicated solvent systems on TLC on silica gel sheets:
(i) ethyl acetate saturated with 0.1M aqueous potassium dihydrogen phosphate, R.sub.f =0.18;
(ii) 3% isopropanol in ethyl acetate saturated with 0.1M aqueous potassium dihydrogen phosphate, R.sub.f =0.28;
(iii) ethyl acetate:
methanol (95:5), R.sub.f =0.27,
e) having a carbon-13 magnetic resonance spectrum as shown in FIG. VIII of the drawings with significant peaks at:
______________________________________ 14.4 17.6 17.9 19.0 19.7 -- 22.8 -- -- 34.0 37.6 39.5 42.1 -- 51.6 52.7 54.1 56.3 57.3 -- 59.3 61.1 61.8 61.9 67.2 68.18 68.23 69.7 70.1 70.8 71.1 71.7 71.8 76.1 -- 81.0 82.9 88.4 -- 97.8 100.0 100.2 101.3 103.0 115.3 123.0 124.9 126.9 130.4 131.1 131.8 138.0 144.7 -- 149.5 149.6 155.6 192.5 192.9 ______________________________________
f) having a molecular formula:
C.sub.55 H.sub.74 N.sub.3 O.sub.21 S.sub.4 Br
g) having a molecular weight:
1319/1321, respectively for .sup.79 Br/.sup.81 Br;
h) and having the structure ##STR8## and which comprises aerobically fermenting the organism Micromonospora echinospora ssp. calichensis NRRL-15839 or mutants thereof including NRRL-15975 and 18149 in a liquid medium containing assimilable sources of carbon, nitrogen, bromine and inorganic salts, until substantial antibiotic activity is imparted to said medium and then recovering the antibiotics therefrom.

69 more elements

A process for producing antibiotics LL-E33288.alpha..sub.1 -Br having the following R.sub.f values in the indicated solvent systems on TLC on silica gel sheets:

a) ethyl acetate saturated with 0.1M aqueous potassium dihydrogen phosphate, R.sub.f =0.67;
b) 3% isopropanol in ethyl acetate saturated with 0.1M aqueous potassium dihydrogen phosphate, R.sub.f =0.80;
and
c) ethyl acetate:
methanol (95:5), R.sub.f =0.79;
LL-E33288.alpha..sub.2 -Br having the following R.sub.f values in the indicated solvent systems on TLC on silica gel sheets:
a) ethyl acetate saturated with 0.1M aqueous potassium dihydrogen phosphate, R.sub.f =0.61;
b) 3% isopropanol in ethyl acetate saturated with 0.1M aqueous potassium dihydrogen phosphate, R.sub.f =0.75;
and
c) ethyl acetate:
methanol (95:5), R.sub.f =0.73;
and having the following structure:
##STR9## LL-E33288.alpha..sub.3 -Br having the following R.sub.f values in the indicated solvent systems on TLC on silica gel sheets:
a) ethyl acetate saturated with 0.1M aqueous potassium dihydrogen phosphate, R.sub.f =0.55;
b) 3% isopropanol in ethyl acetate saturated with 0.1M aqueous potassium dihydrogen phosphate, R.sub.f =0.69;
and
c) ethyl acetate:methanol (95:5), R.sub.f =0.61;
and having the following structure:
##STR10## LL-E33288.alpha..sub.4 -Br having the following R.sub.f values in the indicated solvent systems on TLC on silica gel sheets:
a) ethyl acetate saturated with 0.1M aqueous potassium dihydrogen phosphate, R.sub.f =0.49;
b) 3% isopropanol in ethyl acetate saturated with 0.1M aqueous potassium dihydrogen phosphate, R.sub.f =0.64;
and
c) ethyl acetate:methanol (95:5), R.sub.f =0.54;
LL-E33288.beta..sub.1 -Br
a) having an approximate elemental analysis:
48.6;
H 5.6;
N 2.9;
S 9.1 and Br 5.5;
b) having a melting point:
146.degree.-150.degree. C. (dec.);
c) having a specific rotation:
[.alpha.]D.sup.26 =-49.degree..+-.10.degree. (0.1%, ethanol);
d) having ultraviolet absorption spectra as shown in FIG. I of the drawings;
e) having an infrared absorption spectrum as shown in FIG. II of the drawings;
f) having a proton magnetic resonance spectrum as shown in FIG. III of the drawings;
g) having a carbon-13 magnetic resonance spectrum as shown in FIG. IV of the drawings with significant peaks at:
______________________________________ 17.60(q) 17.64(q) 18.9(q) 19.7(q) 22.4(q) 22.8(q) 23.5(q) 34.3(t) 36.9(t) 39.2(t/d) 47.8(d) 51.7(q) 52.7(q) 54.6(d) 56.3(q) 57.2(q) 57.8(d) 61.0(q/d) 61.7(d) 62.4(t) 66.9(d) 68.4(d) 69.1(d) 69.7(d) 70.2(d) 71.1(d) 71.9(d) 72.1(s) 76.1(d) 81.0(d) 83.3(s) 88.2(s) 97.4(d) 99.7(d) 100.8(s) 102.5(d) 115.1(s) 123.4(d) 124.4(d) 126.5(d) 130.2(s) 130.8(s) 144.6(s) 149.3(s) 149.5(s) 191.7(s) 192.4(s) and ______________________________________
h) having the following R.sub.f values in the indicated solvent systems on TLC on silica gel sheets:
(i) ethyl acetate saturated with 0.1M aqueous potassium dihydrogen phosphate, R.sub.f =0.24;
(ii) 3% isopropanol in ethyl acetate saturated with 0.1M aqueous potassium dihydrogen phosphate, R.sub.f =0.35;
(iii) ethyl acetate:methanol (95:5), R.sub.f =0.36.
i) having a molecular weight:
1333/1335, respectively for .sup.79 Br/.sup.81 Br;
j) having a molecular formula:
C.sub.56 H.sub.76 N.sub.3 O.sub.21 S.sub.4 Br;
k) and having the structure ##STR11## LL-E33288.beta..sub.2 -Br having the following R.sub.f values in the indicated solvent systems on TLC on silica gel sheets:
a) ethyl acetate saturated with 0.1M aqueous potassium dihydrogen phosphate, R.sub.f =0.32;
b) 3% isopropanol in ethyl acetate saturated with 0.1M aqueous potassium dihydrogen phosphate, R.sub.f =0.41;
and
c) ethyl acetate:methanol (95:5), R.sub.f =0.45, and LL-E33288.gamma..sub.1 -Br
a) having ultraviolet absorption spectra as shown in FIG. V of the drawings;
b) having an infrared absorption spectrum as shown in FIG. VI of the drawings;
c) having a proton magnetic resonance spectrum as shown in FIG. VII of the drawings;
and
d) having the following R.sub.f values in the indicated solvent systems on TLC on silica gel sheets:
(i) ethyl acetate saturated with 0.1M aqueous potassium dihydrogen phosphate, R.sub.f =0.18;
(ii) 3% isopropanol in ethyl acetate saturated with 0.1M aqueous potassium dihydrogen phosphate, R.sub.f =0.28;
(iii) ethyl acetate:methanol (95:5), R.sub.f =0.27,
e) having a carbon-13 magnetic resonance spectrum as shown in FIG. VIII of the drawings with significant peaks at:
______________________________________ 14.4 17.6 17.9 19.0 19.7 -- 22.8 -- -- 34.0 37.6 39.5 42.1 -- 51.6 52.7 54.1 56.3 57.3 -- 59.3 61.1 61.8 61.9 67.2 68.18 68.23 69.7 70.1 70.8 71.1 71.7 71.8 76.1 -- 81.0 82.98 88.4 -- 97.8 100.0 100.2 101.3 103.0 115.3 123.0 124.9 126.9 130.4 131.1 131.8 138.0 144.7 -- 149.5 149.6 155.6 192.5 192.9 ______________________________________
f) having a molecular formula:
C.sub.55 H.sub.74 N.sub.3 O.sub.21 S.sub.4 Br
g) having a molecular weight:
1319/1321, respectively for .sup.79 Br/.sup.81 Br;
h) and having the structure ##STR12## which comprises aerobically fermenting a liquid medium containing assimilable sources of carbon, nitrogen, bromine and inorganic salts;
which medium has been inoculated with a viable culture of the organism Micromonospora echinospora ssp. calichensis NRRL-15839 or mutants thereof including NRRL-15975 and 18149 maintaining said fermentation culture at a temperature of about 24.degree.-32.degree. C. for a period of approximately 90-200 hours, harvesting the mash and extracting the antibiotics.

64 more elements

A process for producing antibiotics LL-E33288.alpha..sub.1 -I

a) having the following R.sub.f values in the indicated solvent systems on TLC on silica gel sheets:
(i) ethyl acetate saturated with 0.1M aqueous potassium dihydrogen phosphate, R.sub.f =0.67;
(ii) 3% isopropanol in ethyl acetate saturated with 0.1M aqueous potassium dihydrogen phosphate, R.sub.f =0.80;
(iii) ethyl acetate:methanol (95:5), R.sub.f =0.80;
b) having a molecular weight:
1145.
LL-E33288.alpha..sub.2 -I
a) having the following R.sub.f values in the indicated solvent systems on TLC on silica gel sheets:
(i) ethyl acetate saturated with 0.1M aqueous potassium dihydrogen phosphate, R.sub.f =0.61;
(ii) 3% isopropanol in ethyl acetate saturated with 0.1M aqueous potassium dihydrogen phosphate, R.sub.f =0.75;
and
(iii) ethyl acetate:methanol (95:5), R.sub.f =0.73;
b) containing only the following elements:
C, H, N, O, S and I;
c) having a molecular weight:
1207;
d) having ultraviolet absorption spectra as shown in FIG. IX of the drawings;
e) having an infrared absorption spectrum as shown in FIG. X of the drawings;
f) having a proton magnetic resonance spectrum as shown in FIG. XI of the drawings;
g) having a carbon-13 magnetic resonance spectrum as shown in FIG. XII of the drawings with significant peaks at:
______________________________________ 17.7 56.3 71.8 122.7 18.8 60.7 73.7 125.1 22.7 61.4 79.0 126.3 24.7 60.7 82.6 127.1 33.1 67.7 87.4 133.1 37.4 67.9 85.4 137.0 39.6 69.6 98.0 137.1 41.4 69.7 100.4 149.2 51.3 70.3 100.1 151.5 53.3 71.0 100.8 192.8 53.8 73.4 98.2 193.2 and ______________________________________
h) having a molecular formula:
C.sub.48 H.sub.62 N.sub.3 O.sub.17 S.sub.4 I;
LL-E33288.alpha..sub.3 -I
a) having the following R.sub.f values in the indicated solvent systems on TLC on silica gel sheets:
i) ethyl acetate saturated with 0.1M aqueous potassium dihydrogen phosphate, R.sub.f =0.55;
ii) 3% isopropanol in ethyl acetate saturated with 0.1M aqueous potassium dihydrogen phosphate, R.sub.f =0.69;
and
iii) ethyl acetate:methanol (95:5), R.sub.f =0.61;
b) having a molecular weight:1210;
c) having ultraviolet absorption spectra as shown in FIG. XIII of the drawings;
d) having an infrared absorption spectrum as shown in FIG. XIV of the drawings;
e) having a proton magnetic resonance spectrum as shown in FIG. XV of the drawings;
f) having a carbon-13 magnetic resonance spectrum as shown in FIG. XVI of the drawings with significant peaks at:
______________________________________ 17.5(q) 69.5(d) 103.1(d) 18.0(q) 70.0(d) 123.4(d) 19.0(q) 70.1(d) 124.7(d) 22.7(q) 70.8(d) 127.3(d) 25.3(q) 70.9(d) 131.1 37.4(t) 72.1(s) 130.4(s) 39.2(t) 71.3(d) 133.5(s) 51.5(d) 74.5(d) 136.8(s) 53.3(q) 80.8(d) 143.0(s) 53.7(t) 83.1(s) 145.8 57.2(q) 87.6(s) 150.5(s) 61.0(q) 93.7(s) 151.6(s) 61.8(q) 98.6(s) 154.7(s) 67.1(d) 100.1(d) 192.4(s) 67.3(d) 101.0(s) 192.6(s) 68.0(d) 103.4(d);
and ______________________________________
g) having a molecular formula:
C.sub.47 H.sub.59 N.sub.2 O.sub.19 S.sub.4 I;
LL-E33288.beta..sub.1 -I
a) having the following R.sub.f values in the indicated solvent systems on TLC on silica gel sheets:
(i) ethyl acetate saturated with 0.1M aqueous potassium dihydrogen phosphate, R.sub.f =0.24;
(ii) 3% isopropanol in ethyl acetate saturated with 0.1M aqueous potassium dihydrogen phosphate, R.sub.f =0.35;
and
(iii) ethyl acetate:methanol (95:5), R.sub.f =0.36;
b) having ultraviolet absorption spectra as shown in FIG. XVII of the drawings;
c) having an infrared absorption spectrum as shown in FIG. XVIII of the drawings;
d) having a proton magnetic resonance spectrum as shown in FIG. XIX of the drawings;
e) having a carbon-13 magnetic resonance spectrum as shown in FIG. XX of the drawings with significant peaks at:
______________________________________ -- 17.5 17.6 18.9 -- 22.4 22.8 23.4 25.4 34.3 36.9 39.2 -- 47.9 51.6 52.8 54.8 56.3 57.2 57.9 60.9 -- 61.6 62.2 67.0 68.4 68.4 69.1 69.6 70.4 71.1 71.8 72.2 76.2 -- 80.8 83.3 88.1 93.6 97.4 99.6 99.6 -- 102.6 112.4 123.4 124.4 126.4 -- -- 133.4 -- -- -- -- -- -- 192.2 192.6 -- ______________________________________
f) having a molecular formula:
C.sub.56 H.sub.76 N.sub.3 O.sub.21 S.sub.4 I;
and
g) having a molecular weight:
1381;
LL-E33288.beta..sub.2 -I having the following R.sub.f values in the indicated solvent systems on TLC on silica gel sheets:
a) ethyl acetate saturated with 0.1M aqueous potassium dihydrogen phosphate, R.sub.f =0.32;
b) 3% isopropanol in ethyl acetate saturated with 0.1M aqueous potassium dihydrogen phosphate, R.sub.f =0.41;
and
c) ethyl acetate:methanol (95:5), R.sub.f =0.45;
LL-E33288.gamma..sub.1 -I
a) having the following R.sub.f values in the indicated solvent systems on TLC on silica gel sheets:
(i) ethyl acetate saturated with 0.1M aqueous potassium dihydrogen phosphate, R.sub.f =0.18;
(ii) 3% isopropanol in ethyl acetate saturated with 0.1M aqueous potassium dihydrogen phosphate, R.sub.f =0.28;
and
(iii) ethyl acetate:methanol (95:5), R.sub.f =0.27;
b) containing only the following elements:
C, H, N, O, S and I;
c) having an approximate elemental analysis:
C 48.8;
H 5.4;
N 2.8;
S 9.0;
and I 9.2;
d) having a molecular weight:
1367;
e) having a molecular formula:
C.sub.55 H.sub.74 N.sub.3 O.sub.21 S.sub.4 I;
f) having an ultraviolet absorption spectra as shown in FIG. XXI of the drawings;
g) having an infrared absorption spectrum as shown in FIG. XXII of the drawings;
h) having a proton magnetic resonance spectrum as shown in FIG. XXIII of the drawings;
and
i) having a carbon-13 magnetic resonance spectrum as shown in FIG. XXIV of the drawings, significant peaks as listed below:
______________________________________ 14.5(q) 17.6(q) 17.6(q) 18.9(q) -- -- 22.8(q) -- 25.4(q) 34.1(t) 37.0(t) 39.1(t) 42.3(t/s) -- 51.5(d) 52.8(q) 54.8(t) 56.3(q) 57.2(q) -- 60.4(d) 60.9(q) 61.3(t) 61.7(q) 67.0(d) 68.4(d) 68.5(d) 69.2(d) 69.7(d) 70.5 71.1(d) 71.8(d) 72.1(s) 75.7(d) 75.8(d) 80.9(d) 82.8(s) 88.1(s) 93.5(s) 97.3(d) 99.6(d) 99.7(d) 100.8(s) 102.6(d) -- 123.4(d) 124.4(d) 126.2(d) 103.2(s) 131.0(s) 133.4(s) 139.1(s) 143.0(s) 145.1 150.6(s) 151.5(s) 154.5 192.0(s) 192.5(s) ______________________________________
and LL-E33288.delta.-I
a) having the following R.sub.f values in the indicated solvent systems on TLC on silica gel sheets:
i) ethyl acetate saturated with 0.1M aqueous potassium dihydrogen phosphate, R.sub.f =0.11;
and
ii) 3% isopropanol in ethyl acetate saturated with 0.1M aqueous potassium dihydrogen phosphate, R.sub.f =0.19;
b) having ultraviolet absorption spectra as shown in FIG. XXV of the drawings;
c) having an infrared absorption spectrum as shown in FIG. XXVI of the drawings;
d) having a proton magnetic resonance spectrum as shown in FIG. XXVII of the drawings;
and
e) having a carbon-13 magnetic resonance spectrum as shown in FIG. XXVIII of the drawings, significant peaks as listed below:
______________________________________ 17.2 60.4 99.6 17.6 66.8 103.0 18.5 67.7 122.7 22.4 69.5 124.7 25.0 69.8 126.6 33.0 70.6 130.7 37.3 70.7 130.1 39.0 71.1 133.2 33.5 71.3 137.7 51.0 76.0 142.8 52.4 80.4 150.3 53.6 82.5 151.5 56.1 88.1 155.5 57.0 93.4 192.7 61.0 97.5 192.8 60.6 100.9 61.4 99.8 ______________________________________
which comprises aerobically fermenting the organism Micromonospora echinospora ssp. calichensis NRRL-15839 or mutants thereof including NRRL-15975 and 18149, in a liquid medium containing assimilable sources of carbon, nitrogen, iodine and inorganic salts, until substantial antibiotic activity is imparted to said medium and then recovering the antibiotics therefrom.

92 more elements

A process for producing antibiotics LL-E33288.alpha..sub.1 -I

a) having the following R.sub.f values in the indicated solvent systems on TLC on silica gel sheets:
(i) ethyl acetate saturated with 0.1M aqueous potassium dihydrogen phosphate, R.sub.f =0.67;
(ii) 3% isopropanol in ethyl acetate saturated with 0.1M aqueous potassium dihydrogen phosphate, R.sub.f =0.80;
(iii) ethyl acetate:methanol (95:5), R.sub.f =0.80;
b) having a molecular weight:
1145;
LL-E33288.alpha..sub.2 -I
a) having the following R.sub.f values in the indicated solvent systems on TLC on silica gel sheets:
(i) ethyl acetate saturated with 0.1M aqueous potassium dihydrogen phosphate, R.sub.f =0.61;
(ii) 3% isopropanol in ethyl acetate saturated with 0.1M aqueous potassium dihydrogen phosphate, R.sub.f =0.75;
and
(iii) ethyl acetate:methanol (95:5), R.sub.f =0.73;
b) containing only the following elements:
C, H, N, O, S and I;
c) having a molecular weight:
1207;
d) having ultraviolet absorption spectra as shown in FIG. IX of the drawings;
e) having an infrared absorption spectrum as shown in FIG. X of the drawings;
f) having a proton magnetic resonance spectrum as shown in FIG. XI of the drawings;
g) having a carbon-13 magnetic resonance spectrum as shown in FIG. XII of the drawings with significant peaks at:
______________________________________ 17.7 56.3 71.8 122.7 18.8 60.7 73.7 125.1 22.7 61.4 79.0 126.3 24.7 60.7 82.6 127.1 33.1 67.7 87.4 133.1 37.4 67.9 85.4 137.0 39.6 69.6 98.0 137.1 41.4 69.7 100.4 149.2 51.3 70.3 100.1 151.5 53.3 71.0 100.8 192.8 53.8 73.4 98.2 193.2 and ______________________________________
h) having a molecular formula:
C.sub.48 H.sub.62 N.sub.3 O.sub.17 S.sub.4 I;
LL-E33288.alpha..sub.3 -I
a) having the following R.sub.f values in the indicated solvent systems on TLC on silica gel sheets:
i) ethyl acetate saturated with 0.1M aqueous potassium dihydrogen phosphate, R.sub.f =0.55;
ii) 3% isopropanol in ethyl acetate saturated with 0.1M aqueous potassium dihydrogen phosphate, R.sub.f =0.69;
and
iii) ethyl acetate:methanol (95:5), R.sub.f =0.61;
b) having a molecular weight:
1210;
c) having ultraviolet absorption spectra as shown in FIG. XIII of the drawings;
d) having an infrared absorption spectrum as shown in FIG. XIV of the drawings;
e) having a proton magnetic resonance spectrum as shown in FIG. XV of the drawings;
f) having a carbon-13 magnetic resonance spectrum as shown in FIG. XVI of the drawings with significant peaks at:
______________________________________ 17.5(q) 69.5(d) 103.1(d) 18.0(q) 70.0(d) 123.4(d) 19.0(q) 70.1(d) 124.7(d) 22.7(q) 70.8(d) 127.3(d) 25.3(q) 70.9(d) 131.1 37.4(t) 72.1(s) 130.4(s) 39.2(t) 71.3(d) 133.5(s) 51.5(d) 74.5(d) 136.8(s) 53.3(q) 80.8(d) 143.0(s) 53.7(t) 83.1(s) 145.8 57.2(q) 87.6(s) 150.5(s) 61.0(q) 93.7(s) 151.6(s) 61.8(q) 98.6(s) 154.7(s) 67.1(d) 100.1(d) 192.4(s) 67.3(d) 101.0(s) 192.6(s) 68.0(d) 103.4(d);
and ______________________________________
g) having a molecular formula:
C.sub.47 H.sub.59 N.sub.2 O.sub.19 S.sub.4 I;
LL-E33288.beta..sub.1 -I
a) having the following R.sub.f values in the indicated solvent systems on TLC on silica gel sheets:
(i) ethyl acetate saturated with 0.1M aqueous potassium dihydrogen phosphate, R.sub.f =0.24;
(ii) 3% isopropanol in ethyl acetate saturated with 0.1M aqueous potassium dihydrogen phosphate, R.sub.f =0.35;
and
(iii) ethyl acetate:methanol (95:5), R.sub.f =0.36;
b) having ultraviolet absorption spectra as shown in FIG. XVII of the drawings;
c) having an infrared absorption spectrum as shown in FIG. XVIII of the drawings;
d) having a proton magnetic resonance spectrum as shown in FIG. XIX of the drawings;
e) having a carbon-13 magnetic resonance spectrum as shown in FIG. XX of the drawings with significant peaks at:
______________________________________ -- 17.5 17.6 18.9 -- 22.4 22.8 23.4 25.4 34.3 36.9 39.2 -- 47.9 51.6 52.8 54.8 56.3 57.2 57.9 60.9 -- 61.6 62.2 67.0 68.4 68.4 69.1 69.6 70.4 71.1 71.8 72.2 76.2 -- 80.8 83.3 88.1 93.6 97.4 99.6 99.6 -- 102.6 112.4 123.4 124.4 126.4 -- -- 133.4 -- -- -- -- -- -- 192.2 192.6 -- ______________________________________
f) having a molecular formula:
C.sub.56 H.sub.76 N.sub.3 O.sub.21 S.sub.4 I;
and
g) having a molecular weight:
1381;
LL-E33288.beta..sub.2 -I having the following R.sub.f values in the indicated solvent systems on TLC on silica gel sheets:
a) ethyl acetate saturated with 0.1M aqueous potassium dihydrogen phosphate, R.sub.f =0.32;
b) 3% isopropanol in ethyl acetate saturated with 0.1M aqueous potassium dihydrogen phosphate, R.sub.f =0.41;
and
c) ethyl acetate:methanol (95:5), R.sub.f =0.45;
LL-E33288.gamma..sub.1 -I
a) having the following R.sub.f values in the indicated solvent systems on TLC on silica gel sheets:
(i) ethyl acetate saturated with 0.1M aqueous potassium dihydrogen phosphate, R.sub.f =0.18;
(ii) 3% isopropanol in ethyl acetate saturated with 0.1M aqueous potassium dihydrogen phosphate, R.sub.f =0.28;
and
(iii) ethyl acetate:methanol (95:5), R.sub.f =0.27;
b) containing only the following elements:
C, H, N, O, S and I;
c) having an approximate elemental analysis:
C 48.8;
H 5.4;
N 2.8;
S 9.0;
and I 9.2;
d) having a molecular weight:
1367;
e) having a molecular formula:
C.sub.55 H.sub.74 N.sub.3 O.sub.21 S.sub.4 I;
f) having an ultraviolet absorption spectra as shown in FIG. XXI of the drawings;
g) having an infrared absorption spectrum as shown in FIG. XXII of the drawings;
h) having a proton magnetic resonance spectrum as shown in FIG. XXIII of the drawings;
and
i) having a carbon-13 magnetic resonance spectrum as shown in FIG. XXIV of the drawings, significant peaks as listed below:
______________________________________ 14.5(q) 17.6(q) 17.6(q) 18.9(q) -- -- 22.8(q) -- 25.4(q) 34.1(t) 37.0(t) 39.1(t) 42.3(t/s) -- 51.5(d) 52.8(q) 54.8(t) 56.3(q) 57.2(q) -- 60.4(d) 60.9(q) 61.3(t) 61.7(q) 67.0(d) 68.4(d) 68.5(d) 69.2(d) 69.7(d) 70.5 71.1(d) 71.8(d) 72.1(s) 75.7(d) 75.8(d) 80.9(d) 82.8(s) 88.1(s) 93.5(s) 97.3(d) 99.6(d) 99.7(d) 100.8(s) 102.6(d) -- 123.4(d) 124.4(d) 126.2(d) 103.2(s) 131.0(s) 133.4(s) 139.1(s) 143.0(s) 145.1 150.6(s) 151.5(s) 154.5 192.0(s) 192.5(s) ______________________________________
LL-E33288.delta.-I
a) having the following R.sub.f values in the indicated solvent systems on TLC on silica gel sheets;
(i) ethyl acetate saturated with 0.1M aqueous potassium dihydrogen phosphate, R.sub.f =0.11;
and
(ii) 3% isopropanol in ethyl acetate saturated with 0.1M aqueous potassium dihydrogen phosphate, R.sub.f =0.19;
b) having ultraviolet absorption spectra as shown in FIG. XXV of the drawings;
c) having an infrared absorption spectrum as shown in FIG. XXVI of the drawings;
d) having a proton magnetic resonance spectrum as shown in FIG. XXVII of the drawings;
and
e) having a carbon-13 magnetic resonance spectrum as shown in FIG. XXVIII of the drawings, significant peaks as listed below:
______________________________________ 17.2 60.4 99.6 17.6 66.8 103.0 18.5 67.7 122.7 22.4 69.5 124.7 25.0 69.8 126.6 33.0 70.6 130.7 37.3 70.7 130.1 39.0 71.1 133.2 33.5 71.3 137.7 51.0 76.0 142.8 52.4 80.4 150.3 53.6 82.5 151.5 56.1 88.1 155.5 57.0 93.4 192.7 61.0 97.5 192.8 60.6 100.9 61.4 99.8 ______________________________________
which comprises aerobically fermenting a liquid medium containing assimilable sources of carbon, nitrogen, iodine and inorganic salts, which medium has been inoculated with a viable culture of the microorganism Micromonospora echinospora ssp. calichensis NRRL-15839 or mutants thereof including NRRL-15975 and 18149, maintaining said fermentation culture at a temperature of about 24.degree.-32.degree. C. for a period of approximately 90-200 hours, harvesting the mash and extracting the antibiotics.

93 more elements

Dependencies are inferred from the claim text.

Facts

Application: 07/560,415Filed 10 Sep 1990 · division
Priority: 16 Nov 1984
Granted: 28 Apr 1992
Term ends: Not estimatedThe source records don’t support an estimate.

Classification11 codes

C07H 15/203
Carbocyclic rings: monocyclic carbocyclic rings other than cyclohexane rings; Bicyclic carbocyclic ring systems
C12N 1/205
Bacteria; Culture media therefor: bacterial isolates
C12P 19/60
Preparation of O-glycosides, e.g. glucosides: having an oxygen of the saccharide radical directly bound to a non-saccharide heterocyclic ring or a…
C12R 2001/29
Bacteria or Actinomycetales ; using bacteria or Actinomycetales: micromonospora

Family

Serial672,031Parent of this continuation-in-part · filed 16 Nov 1984
Serial787,066Parent of this continuation-in-part · filed 17 Oct 1985
Serial9,321Parent of this division · filed 30 Jan 1987
US 4,970,198 AParent of this division
US 5,108,912 AThis patent

From the US filing history. Foreign family members aren’t listed.

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Abstract

A process for producing antibiotics LL-E33288.alpha..sub.1 -Br having the following R.sub.f values in the indicated solvent systems on TLC on silica gel sheets:

a) ethyl acetate saturated with 0.1M aqueous potassium dihydrogen phosphate, R.sub.f =0.67;
b) 3% isopropanol in ethyl acetate saturated with 0.1M aqueous potassium dihydrogen phosphate, R.sub.f =0.80;
and
c) ethyl acetate:
methanol (95:5), R.sub.f =0.79;
LL-E33288.alpha..sub.2 -Br having the following R.sub.f values in the indicated solvent systems on TLC on silica gel sheets:
a) ethyl acetate saturated with 0.1M aqueous potassium dihydrogen phosphate, R.sub.f =0.61;
b) 3% isopropanol in ethyl acetate saturated with 0.1M aqueous potassium dihydrogen phosphate, R.sub.f =0.75;
and
c) ethyl acetate:
methanol (95:5), R.sub.f =0.73;
and having the following structure:
##STR5## LL-E33288.alpha..sub.3 -Br having the following R.sub.f values in the indicated solvent systems on TLC on silica gel sheets:
a) ethyl acetate saturated with 0.1M aqueous potassium dihydrogen phosphate, R.sub.f =0.55;
b) 3% isopropanol in ethyl acetate saturated with 0.1M aqueous potassium dihydrogen phosphate, R.sub.f =0.69;
and
c) ethyl acetate:
methanol (95:5), R.sub.f =0.61;
and having the following structure:
##STR6## LL-E33288.alpha..sub.4 -Br having the following R.sub.f values in the indicated solvent systems on TLC on silica gel sheets:
a) ethyl acetate saturated with 0.1M aqueous potassium dihydrogen phosphate, R.sub.f =0.49;
b) 3% isopropanol in ethyl acetate saturated with 0.1M aqueous potassium dihydrogen phosphate, R.sub.f =0.64;
and
c) ethyl acetate:
methanol (95:5), R.sub.f =0.54;
LL-E33288.beta..sub.1 -Br
a) having an approximate elemental analysis:
C 48.6;
H 5.6;
N 2.9;
S 9.1 and Br 5.5;
b) having a melting point:
146.degree.-150.degree. C. (dec.);
c) having a specific rotation:
[.alpha.]D.sup.26 =-49.degree..+-.10.degree. (0.1%, ethanol);
d) having ultraviolet absorption spectra as shown in FIG. I of the drawings;
e) having an infrared absorption spectrum as shown in FIG. II of the drawings;
f) having a proton magnetic resonance spectrum as shown in FIG. III of the drawings;
g) having a carbon-13 magnetic resonance spectrum as shown in FIG. IV of the drawings with significant peaks at:
______________________________________ 17.60(q) 17.64(q) 18.9(q) 19.7(q) 22.4(q) 22.8(q) 23.5(q) 34.3(t) 36.9(t) 39.2(t/d) 47.8(d) 51.7(q) 52.7(q) 54.6(d) 56.3(q) 57.2(q) 57.8(d) 61.0(q/d) 61.7(d) 62.4(t) 66.9(d) 68.4(d) 69.1(d) 69.7(d) 70.2(d) 71.1(d) 71.9(d) 72.1(s) 76.1(d) 81.0(d) 83.3(s) 88.2(s) 97.4(d) 99.7(d) 100.8(s) 102.5(d) 115.1(s) 123.4(d) 124.4(d) 126.5(d) 130.2(s) 130.8(s) 144.6(s) 149.3(s) 149.5(s) 191.7(s) 192.4(s) and ______________________________________
h) having the following R.sub.f values in the indicated solvent systems on TLC on silica gel sheets:
(i) ethyl acetate saturated with 0.1M potassium dihydrogen phosphate, R.sub.f =0.24;
(ii) 3% isopropanol in ethyl acetate saturated with 0.1M aqueous potassium dihydrogen phosphate, R.sub.f =0.35;
(iii) ethyl acetate:
methanol (95:5), R.sub.f =0.36.
i) having a molecular weight:
1333/1335, respectively for .sup.79 Br/.sup.81 Br;
j) having a molecular formula:
C.sub.56 H.sub.76 N.sub.3 O.sub.21 S.sub.4 Br;
k) and having the structure ##STR7## LL-E33288.beta..sub.2 -Br having the following R.sub.f values in the indicated solvent systems on TLC on silica gel sheets:
a) ethyl acetate saturated with 0.1M aqueous potassium dihydrogen phosphate, R.sub.f =0.32;
b) 3% isopropanol in ethyl acetate saturated with 0.1M aqueous potassium dihydrogen phosphate, R.sub.f =0.41;
and
c) ethyl acetate:
methanol (95:5), R.sub.f =0.45;
and LL-E33288.gamma..sub.1 -Br
a) having ultraviolet absorption spectra as shown in FIG. V of the drawings;
b) having an infrared absorption spectrum as shown in FIG. VI of the drawings;
c) having a proton magnetic resonance spectrum as shown in FIG. VII of the drawings;
and
d) having the following R.sub.f values in the indicated solvent systems on TLC on silica gel sheets:
(i) ethyl acetate saturated with 0.1M aqueous potassium dihydrogen phosphate, R.sub.f =0.18;
(ii) 3% isopropanol in ethyl acetate saturated with 0.1M aqueous potassium dihydrogen phosphate, R.sub.f =0.28;
(iii) ethyl acetate:
methanol (95:5), R.sub.f =0.27,
e) having a carbon-13 magnetic resonance spectrum as shown in FIG. VIII of the drawings with significant peaks at:
______________________________________ 14.4 17.6 17.9 19.0 19.7 -- 22.8 -- -- 34.0 37.6 39.5 42.1 -- 51.6 52.7 54.1 56.3 57.3 -- 59.3 61.1 61.8 61.9 67.2 68.18 68.23 69.7 70.1 70.8 71.1 71.7 71.8 76.1 -- 81.0 82.9 88.4 -- 97.8 100.0 100.2 101.3 103.0 115.3 123.0 124.9 126.9 130.4 131.1 131.8 138.0 144.7 -- 149.5 149.6 155.6 192.5 192.9 ______________________________________
f) having a molecular formula:
C.sub.55 H.sub.74 N.sub.3 O.sub.21 S.sub.4 Br
g) having a molecular weight:
1319/1321, respectively for .sup.79 Br/.sup.81 Br;
h) and having the structure ##STR8## and which comprises aerobically fermenting the organism Micromonospora echinospora ssp. calichensis NRRL-15839 or mutants thereof including NRRL-15975 and 18149 in a liquid medium containing assimilable sources of carbon, nitrogen, bromine and inorganic salts, until substantial antibiotic activity is imparted to said medium and then recovering the antibiotics therefrom.

69 more elements

Facts

Application

07/560,415Filed 10 Sep 1990 · division

Priority

16 Nov 1984

Granted

28 Apr 1992

Term ends

Not estimatedThe source records don’t support an estimate.

Classification11 codes

C07H 15/203
Carbocyclic rings: monocyclic carbocyclic rings other than cyclohexane rings; Bicyclic carbocyclic ring systems
C12N 1/205
Bacteria; Culture media therefor: bacterial isolates
C12P 19/60
Preparation of O-glycosides, e.g. glucosides: having an oxygen of the saccharide radical directly bound to a non-saccharide heterocyclic ring or a…
C12R 2001/29
Bacteria or Actinomycetales ; using bacteria or Actinomycetales: micromonospora

Family

Serial672,031Parent of this continuation-in-part · filed 16 Nov 1984

Serial787,066Parent of this continuation-in-part · filed 17 Oct 1985

Serial9,321Parent of this division · filed 30 Jan 1987

US 4,970,198 AParent of this division

US 5,108,912 AThis patent

From the US filing history. Foreign family members aren’t listed.