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US 4,486,414 AGranted
Dolastatins A and B cell growth inhibitory substances
AssigneeArizona Board of Reagents
InventorGeorge R. Pettit
Life of this patent
Estimating the term…
Abstract
Potent cell growth inhibitory substances have been obtained from the Indian Ocean sea hare Dolabella. These substances have been given the names dolastatin A and dolastatin B. These compounds are characterized by physical and chemical parameters.
Claims7
Cell growth inhibitory substance designated dolastatin A which has the following characteristics:
- Physical Data for Dolastatin A
- ______________________________________ TLC, silica gel:
- Rf Solvent Ratio ______________________________________ 0.53 CH.sub.2 Cl.sub.2 /MeOH/H.sub.2 O 90:10:0.8 0.49 EtOAc/EtOH 80:20 0.33 EtOAc/EtOH 90:10 0.15 CH.sub.2 Cl.sub.2 /MeOH/H.sub.2 O + 5% HOAc 90:10:0.8 0.68 CH.sub.2 Cl.sub.2 /MeOH/H.sub.2 O - 5% Et.sub.2 NH 90:10:0.8 ______________________________________ TLC, silica gel visualization:
- UV ++ H.sub.2 SO.sub.4 - Anisaldehyde pale yellow - Ninhydrin Dragendorf orange, before heating - after heating Phosphomolibdic + acid ______________________________________
- IR, KBr:
- 3430, 2964, 2938, 2874, 1731, 1663, 1630, 1446, 1379, 1308, 1252, 1192,
- UV (MeOH):
- .lambda..sub.max 202, 204, 206 nm (E 8248), .lambda..sub.max 237 nm (E 3461).
- Elemental Analysis:
- C, 63.34;
- H, 10.06;
- N, 12.10;
- S, 5.64;
- Amino Acid Analysis:
- ______________________________________ Hydrolysis by:
- H.sup.+ OH.sup.- ______________________________________ Proline (16.61%) Proline (11.63%) Valine (4.99%) Leucine (10.49%) Leucine (8.63%) no Valine was found + one unidentified compound ______________________________________
- MS:
- FAB:
- 837 (M.sup.+ +H), 552 (M.sup.+ +H-285), 498 (M.sup.+ +H-339), 340 (M.sup.+ +H-497), 297, 295, 288, 286, 258, 227, 215, 213, 207, 204, 199, 197, 100 (C.sub.6 H.sub.14 N.sup.+), 86, 72, 70, 45;
- GCMS:
- Valine, Leucine and Proline identified;
- NMR:
- 400 MH.sub.z, CDCl.sub.3, .delta.:
- ______________________________________ 0.763 (3H, d, J=6.56H.sub.z) 3.732 (3H, S) 0.908 (.about.3H, d, J=7.16) 3.775 (.about.2H, dd, J=4.56, 14.68) 0.926 (.about.6H, d, J=7.64) 3.877 (1H, broad t, J=?) 1.019 (3H, d, J=6.44) 4.618 (1H, t, J=.about.6.10) 1.057 (3H, d, J=6.84) 4.699 (1H, S) 1.232 (?, impurity, .about.1H, S) 4.756 (1H, t, J=3.72) 1.294 (1H, t, J=7.00, impurity?) 4.818 (1H, dd, J=2.46, 8.70) 1.558 (7-8H, broad S, H.sub.2 O?) 5.115 (1Hd, J=11.04) 1.845 (1H, td, J=11.76, 5.88) 5.282 (2-3H, S) 2.02-2.03 (.about.3H, m) 5.869 (1H, d, J=2.48) 2.12-2.14 (.about.2H, m .about.5.9 (1H, broad peak) 2.05-2.3 (.about.11H, m) 712-7.21 (.about.5H, m) .about.2.35 (.about.1H, m) 3.026 (1H, dd, J=3.32, 13.92) 3.157 (3H, S) 3.513 (1H, dd, J=14.12, 4.36 3.581 (1H, dd, J=7.16, ?). ______________________________________
21 more elements
Cell growth inhibitory substance designated dolastatin B which has the following characteristics:
- Physical Data for Dolastatin B
- ______________________________________ TLC, silica gel:
- Rf Solvent Ratio ______________________________________ 0.14 CH.sub.2 Cl.sub.2 :MeOH:H.sub.2 O 90:10:0.8 .about.0. EtOAc:EtOH 80:20 .about.0. EtOAc:EtOH 90:10 0.33 CH.sub.2 Cl.sub.2 :MeOH:H.sub.2 O + 5% HOAc 90:10:0.8 0.54 CH.sub.2 Cl.sub.2 :MeOH:H.sub.2 O + 5% Et.sub.2 NH 90:10:0.8 ______________________________________ Visualization on TLC (silica gel):
- ______________________________________ UV ++ H.sub.2 SO.sub.4 - Anisaldehyde pale yellow Ninhydrin - Dragendorf orange before heating - after heating Phosphomolibdic + acid ______________________________________
- IR, KBr:
- 3433, 2963, 2925, 2873, 2852, 1729, 1668, 1661, 1630, 1567, 1445, 1383, 1306, 1253, 1190,
- UV (MeOH):
- .lambda..sub.max 203, 206, nm (E 8332), .lambda..sub.max 238 nm (E 3525),
- MS:
- FAB:
- 853 (M.sup.+ +H, (837), 823, (611), (583), 568, (552), 498, 356, 340, 326, 297, 296, 295, 288, 286, 281, (277?), 274, (243), 215, 213, 204 (197), (185?), 182, (154), 144, (131), (115?, 114, 112), 100, 98, (93!), 91, 86, 84, 83, (75?), 72, 70, 69, (61), (57?), 55, 45, 43, 41, 39, 31, 29, m/e 86 C.sub.5 H.sub.12 N, m/e 100 C.sub.6 H.sub.14 N,
- NMR:
- 400 MH.sub.z, CDCl.sub.3, .delta.:
- ______________________________________ 0.7715 (3H, d, J=6.28H.sub.z) 3.4728 (1H, S) 0.8588 (3H, t, J=6.70H.sub.z) 3.527 (1H, dd, J=14.4 and 5.2H.sub.z) 0.9076 (3H, d, J=7.04H.sub.z) 3.602 (1H, m) 0.9350 (3H?, d, J=6.36H.sub.z) 3.665 (?) 0.9864 (3H, S?) .about.3.72 (1H, m) 1.0392 (3H, d, J=6.80H.sub.z) 3.733 (3H, S) 1.0567 (3H, d, J=7.16H.sub.z) 3.800 (14, dd, J=?) 1.295.about.1.315 (.about.1.about.2H, m) 3.89 (1H, broad t, J=?) 1.405 (.about.1H, S) impurity? 4.62 (1H, broad t, J=?) 1.79-1.90 (1.about.2H) 4.701 (1H, S) 2.02-2.06 (22-26H) 4.757 (1H, t or d) 2.10-2.22 4.825 (1H, dd, J=8.8 and ? H.sub.z) 2.22-2.27 5.125 (1H, d, J=12H.sub.z) 2.32-2.40 5.281 (.about.9H, S, ?, CH.sub.2 Cl.sub.2 ?) .about.2.55 (.about.2H, broad peak) 5.876 (1H, d, J=?) 3.043 (1H, dd, J=14.4 and 3.3H.sub.z) 7.11-7.22 (4-5H, m) 3.152 (3H, S) .about.7.75 (1H, broad peak) ______________________________________
11 more elements
A process for treating a host having P388 leukemia which comprises the administration of an effective amount of dolastatin A to said host.
A process for treating a host having P388 leukemia which comprises the administration of an effective amount of dolastatin B to said host.
A process for preparing dolastatins active against P388 leukemia which comprises:
- (a) extracting a ground preparation of the Indian Ocean sea hare Dolabella wth methylene chloride:methanol (1:1) to obtain an extract;
- (b) subjecting said extract to solvent partitioning to obtain an extract showing activity against P388 leukemia;
- (c) chromatographing said active extract and isolating the fractions active against P388 leukemia;
- and,
- (d) rechromatographing said active fraction to obtain further fractions active against P388 leukemia.
2 more elements
Dependencies are inferred from the claim text.