Transformations of codeine to important semisynthetic opiate derivatives by Pseudomonas putida m10

Abstract

A biotransformation mixture which contained codeine and washed cells of Pseudomonas putida M10 gave rise to a number of transformation products that are of clinical importance which included hydrocodone, dihydrocodeine and 14L-hydroxycodeine. Incubations with the same organism and codeinone gave rise to 14L-hydroxycodeinone and 14L-hydroxycodeine. Cell-free extracts and membrane fractions of P. putida M10 were shown to catalyse the 14L-hydroxylation of codeinone. In addition, the potent analgesic oxycodone was shown to be produced from 14L-hydroxycodeinone. ß

The potential of alkaloids in drug discovery

“…These biotransformations may have important implications for the production of compounds of pharmaceutical significance from more readily available precursors when synthetic conversions are low yielding (Lister et al, 1999). When such enzymes have broader substrate specificity, synthetic transformations may also be achieved with systems that were originally discovered based on non-alkaloidal pathways.…”

Bioconversion of codeine to semi-synthetic opiate derivatives by the cyanobacterium Nostoc muscorum

“…The second step, on the other hand, seems to be a chemical rather than enzymatic event, as no enzyme that could catalyze this reaction has been identified so far (Zhang et al 2005). It is of interest to note that unlike most previously studied microorganisms (Kunz et al 1985;Liras and Umbreit 1975;Lister et al 1999;Zhang et al 2005), the major product of 14-hydroxylation of codeine by N. muscorum is oxycodone, indicating that other enzymes are also involved. Lister and co-workers have demonstrated that purified morphinone reductase from P. putida catalyzes the transformation of 14-hydroxycodeinone to oxycodone in vitro, although they could not detect oxycodone in the biotransformation mixture containing live P. putida cells.…”

“…Among these, C14-hydroxylation is of particular importance, since it has been shown to improve the pharmacological properties of morphine alkaloids (Lister et al 1999;Zhang et al 2005). Generally, 14-hydroxylated opioids are synthesized using thebaine as the starting material.…”

“…However, thebaine constitutes only a small fraction of natural opium extracts. For this reason, considerable effort has been directed toward developing efficient biocatalytic processes for the 14-hydroxylation of morphine alkaloids with tertiary carbon at C-14, and some microorganism with the ability to perform such modifications, including Pseudomonas putida (Lister et al 1999) and Mycobacterium neoaurum (Zhang et al 2005), have been identified. The microbial transformation of morphine alkaloids to their corresponding 17-hydroxy derivatives appears to be a multistep process involving C6-oxidation, C14-hydroxylation, and C6-reduction (Zhang et al 2005).…”

Cytochrome P450 3A Enzymes Catalyze the O6-Demethylation of Thebaine, a Key Step in Endogenous Mammalian Morphine Biosynthesis

“…Previous attempts to characterize the enzymes that catalyze O 6 -demethylation of thebaine and oripavine have been unsuccessful, in large part due to instability of codeinone and morphinone in aqueous solution (34,35). In the present work, we devised and validated a coupled enzyme assay to circumvent this issue.…”