Fermentative preparation process for and crystal forms of cytostatics
OwnerHans Hofmann, Marion Mahnke +5inventors
InventorsHans Hofmann, Marion Mahnke, Klaus Memmert, Frank Petersen, Thomas Schupp, Ernst Kusters, Michael Mutz
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Abstract
The invention relates to a new process for concentrating epothilones in culture media, a new process for the production of epothilones, a new process for separating epothilones A and B and a new strain obtained by mutagenesis for the production of epothilones, as well as aspects related thereto. New crystal forms of epothilone B are also described.
Claims23
A process for concentrating epothilones in a culture medium for the biotechnological preparation of these compounds, the process comprising microorganisms which produce these compounds, whereby a complex-forming component which is soluble in the culture medium is added to the medium.
A culture medium which comprises a complex-forming component and a microorganism suitable for the production of epothilones.
A process for the production of epothilones, in which the epothilones are obtained by working up a culture medium for the biotechnological preparation of these compounds, the medium comprising myxobacteria as producers of natural substances, to which medium is added a complex-forming component which is soluble in the culture medium, and subsequently purifying and, if desired, separating the epothilones from one another.
A process for the production of epothilones, which
- a) is a process for concentrating epothilones in a culture medium for the biotechnological preparation of these compounds, which contains a microorganism suitable for the preparation thereof, water and other suitable customary constituents of culture media, whereby a cyclodextrin or a cyclodextrin derivative is added to the medium, or a mixture of two or more of these compounds;
- and
- b) comprises a step for separating epothilones from one another, which is characterised by chromatography on a reversed-phase column with an eluant containing a lower alkyl-cyanide, whereby chromatography is carried out on column material charged with hydrocarbon chains, and an eluant containing a lower alkylnitrile is used;
- whereby if desired further working up steps and purification steps are possible.
A method of separating epothilones from one another, which is characterised by chromatography on a reversed-phase column with an eluant containing a lower alkyl-cyanide.
A strain of Sorangium cellulosum, obtained by mutagenesis, which under otherwise identical conditions, produces more epothilones than Sorangium cellulosum Soce90.
A crystal form of epothilone B having the reference modification A, which is characterised by the X-ray diffraction diagram reproduced in the form of a table, obtained using a diffractometer with Cu-Kα1 radiation.
- 16 2θ Intensity 7.7 very strong 10.6 weak 13.6 average 14.4 average 15.5 average 16.4 weak 16.8 weak 17.1 weak 17.3 weak 17.7 weak 18.5 weak 20.7 strong 21.2 strong 21.9 weak 22.4 weak 23.3 strong 25.9 average 31.2 weak 32.0 average
A crystal form of epothilone B having the reference modification B, which is characterised by the X-ray diffraction diagram reproduced in the form of a table, obtained using a diffractometer with Cu-Kα1 radiation.
- 17 2θ Intensity 6.9 very strong 8.0 weak 8.3 average 10.8 strong 11.5 average 12.4 weak 13.1 strong 15.5 weak 16.2 weak 16.7 average 18.1 average 18.6 average 20.4 weak 20.9 strong 21.3 weak 21.5 very weak 22.5 average 24.2 weak 25.1 average
Dependencies are inferred from the claim text.