Mechanism of the solution oxidation of rofecoxib under alkaline conditions.
OBJECTIVE
The rapid oxidation of rofecoxib under alkaline conditions has been previously reported. The oxidation was reported to involve gamma-lactone ring opening to an alcohol, which further oxidized to a dicarboxyclic acid. The oxidation was suspected to be mediated by peroxy radicals. This work further investigates the mechanism of oxidation under the alkaline solution conditions.


METHODS
The pH dependence of the oxidation reaction was determined in 50% acetonitrile/50% aqueous phosphate buffer (pH 9-12). The oxidation reaction products were also examined at early timepoints (from 40 s to several minutes) with only 5% water content. The evolution of hydrogen peroxide by the oxidation reaction was quantitatively followed by reaction with triphenylphosphine (TPP) and high-pressure liquid chromatography determination of the resultant triphenylphosphine oxideformed. Rofecoxib was exposed to the alkaline pH conditions in the presence of formaldehyde, and the primary reaction product was isolated and characterized by liquid chromatography-mass spectrometry and proton 1D, heteronuclear multiple quantum coherence (HMQC), gradient heteronuclear multiple bond correlation (gHMBC), and carbon 1D nuclear magnetic resonance techniques. Transient reaction products were examined for hydroperoxide groups by reaction with TPP.


RESULTS
The oxidation reaction occurs only near pH 11 and above. In the presence of excess formaldehyde, oxidation products are no longer observed but a new product is observed in which two formaldehyde molecules have added to the methylene carbon atom of the gamma-lactone ring. The evolution of hydrogen peroxide corresponds quantitatively to the molar amount of the (minor) aldehyde oxidation product formed. It is demonstrated that the rofecoxib anhydride species is actually the primary product of the oxidation reaction. The existence of a transient hydroperoxide species is shown by reaction with TPP and concomitant conversion to a previously identified alcohol.


CONCLUSIONS
The oxidation of rofecoxib under these high pH conditions is mediated by rofecoxib enolate ion formation. The enolate ion reacts with either formaldehyde or dissolved oxygen at the C5 position. In the case of oxygen, a transient hydroperoxide species is formed. The major and minor products of the oxidation derive from competitive routes of decomposition of this hydroperoxide. The major route involves a second enolate ion formation, which decomposes with heterolytic cleavage of the RO-OH bond to give the rofeocoxib anhydride and hydroxide ion. The anhydride is rapidly hydrolyzed under the alkaline conditions to give the observed rofecoxib dicarboxylate product. The minor hydroxy-furanone product is formed from hydroxide ion attack on the hydroperoxide intermediate.
Harmon
Paul A
PA
Department of Pharmaceutical Analysis and Control, Merck Research Laboratories, P.O. Box 4, West Point, Pennsylvania, 19486, USA. Paul_Harmon@Merck.com
Biffar
Stephen
S
Pitzenberger
Steven M
SM
Reed
Robert A
RA
D000468:Alkalies / Q000737:chemistry*; D004355:Drug Stability; D005663:Furans / Q000737:chemistry; D006861:Hydrogen Peroxide / Q000378:metabolism; D006863:Hydrogen-Ion Concentration; D007783:Lactones / Q000737:chemistry* / Q000378:metabolism; D010084:Oxidation-Reduction; D012996:Solutions; D013450:Sulfones / Q000737:chemistry* / Q000378:metabolism; D013997:Time Factors
D000468:Alkalies; D005663:Furans; D007783:Lactones; D012996:Solutions; D013450:Sulfones; C116926:rofecoxib; D006861:Hydrogen Peroxide