Morphinone reductase for the preparation of hydromorphone and hydrocodone
AssigneeMacfarlan Smith Limited
InventorsAnne M. Hailes, Christopher E. French, Neil C. Bruce
Life of this patent
Estimating the term…
Abstract
A morphinone reductase enzyme and related process provide for the preparation of hydromorphone from morphinone and hydrocodone from neopinone or codeinone. The enzyme has no significant activity with compounds of similar ring structure, has a molecular weight of approximately 81 kilodaltons and is composed of two identical subunits. The enzyme uses NADH as a cofactor and displays no significant enzymatic activity in the reverse direction.
Claims9
A purified morphinone reductase enzyme having the N-terminal amino-acid sequence:
- Pro Asp Thr Ser Phe Ser Asn Pro Gly Leu Phe Thr Pro Leu Gln (Seq ID NO:1).
A purified morphinone reductase enzyme wherein:
- (1) with NADH as cofactor said enzyme reduces morphinone to hydromorphone;
- (2) with said cofactor said enzyme reduces codeinone and neopinone but has essentially no enzymatic activity in reverse direction;
- (3) said enzyme has a molecular weight of about 81,000, as determined by gel filtration;
- and
- (4) in phosphate buffer said enzyme exhibits maximal activity at pH about 8.0.
A process of producing an enzyme, said enzyme having morphinone reductase activity, said activity comprising a capacity for reducing morphinone to hydromorphone, said process comprising:
- culturing an enzyme obtainable from the strain Pseudomonas putida, NCIMB 40119 or a mutant or variant thereof capable of producing a morphinone reductase enzyme which, with a cofactor, reduces morphinone to hydromorphone, wherein the culturing is conducted on a source of carbon and nitrogen, at a temperature of 20.degree. to 40.degree. C., until the morphinone reductase is produced disrupting the cells;
- identifying a fraction having said morphinone reductase enzyme activity;
- and recovering said morphinone reductase enzyme having said morphinone reductase activity.
Dependencies are inferred from the claim text.