Microbiological hydroxylation process of quinine, quinidine and

Chemistry: molecular biology and microbiology – Micro-organism – tissue cell culture or enzyme using process... – Preparing heterocyclic carbon compound having only o – n – s,...

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435122, 435931, C12P 1716, C12P 1712, C12N 114

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active

049961497

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BRIEF SUMMARY
This invention relates to a process for the hydroxylation of cinchona alkaloids, and more particularly to a new process for hydroxylation of quinine, quinidine, and their derivatives, by microbiological means.
Hepatic microsomal hydroxylation frequently constitutes one of the first stages of the metabolism of a foreign chemical compound introduced in a mammal's alimentary or blood circulation systems. Such hydroxylations, followed by other reactions, such as O-methylation or conjugation, are designed to render those substances more water-soluble, and therefore to accelerate their elimination. However, in certain cases, the hydroxylated compounds are more active, or more toxic, than the original molecule; this justifies interest in their identification, and therefore their preparation with a view towards the study of their activity.
Organic synthesis methods are obviously convenient for metabolites of simple substances, but the the chemical preparation of more complex (and optically active) molecules can require substantial effort and time.
Hydroxylations by microbiological means have already been developed considerably for many years in the field of steroids, where they have enabled obtaining, on an industrial scale, regio- and steroselectively hydroxylated products, resulting from specific oxidation of non functionalized carbon atoms. Such molecules would have required considerable synthesis efforts had it been necessary to utilize chemical methods. This method has since been applied to other families of molecules and it constitutes, in certain cases, a preferred way of obtaining new compounds, transposable to large scale production by utilization of classical fermentation techniques and equipment. Furthermore, it appears that microbic hydroxylations lead for each species to a frequently narrower range of metabolites than found in the mammals' systems, thus eliminating some separation problems; it is then sufficient to take advantage of the multiplicity of strains to reconstitute the various possibilities of hepatic metabolites.
However, in the field of alkaloids, rather few reactions of the said type have been described, and they relate in most cases to the non-nitrogenated part of the aromatic or non steroidic molecule. It is therefore both desirable and interesting to be able to carry out, in favourable conditions, the hydroxylation of nitrogenous alkaloid nuclei whose products have already been recognized as active metabolites. In particular, French patent No. 2,506,312 describes hydoxylated compounds of quinidine, such as hydroxy-3 dihydro-10,11 quinidine and its 3R and 3S epimers, which can be utilized therapeutically for the treatment of cardiac arrythmias. On the other hand, we are aware of the fact that quinine, whose pharmacological properties are known, and quinidine are two isomers, which differ in their configurations at the level of the carbons in positions 8 and 9.
The preparation of such active, optically pure, compounds, by a microbiological method, transposable to industrial scaling-up, therefore represents an objective of obvious interest as regards chemical hydroxylations, which lead to low yields and diastereoisomeric mixtures.
The object of this invention is therefore to provide a new process for microbiological hydroxylation of quinine, quinidine, and their derivatives, in particular their hydrogenated derivatives, in a regio- and stereospecific manner, so as to obtain optically pure compounds.
Another object of this invention is to provide a process of microbiolgical hydroxylation of quinine, quinidine, and their derivatives, susceptible of industrial application, and to supply optically pure compounds, with a satisfactory yield, by means of apparatus of conventional type.
The regiospecific and stereospecific microbiolgical hydroxylation process according to this invention allows hydroxylation, in the 3S position, of quinine, quinidine, and their derivatives, represented by formula (I): ##STR1## in which R represents an ethyl or vinyl group, to form the corresponding 3S-hydroxylate

REFERENCES:
Eckenrode, F. M., "Microbiological Transformation of Quinidine", Journal of Natural Products, vol. 47, No. 5, (Sep.-Oct., 1984) pp. 882-884.

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