Munir N. Nassar

Influence of Formaldehyde Impurity in Polysorbate 80 and PEG‐300 on the Stability of a Parenteral Formulation of BMS‐204352: Identification and Control of the Degradation Product

The purpose of this study was to identify a degradation product formed in the clinical parenteral formulation of BMS‐204352, investigate the role of excipients in its formation, and develop a strategy to minimize/control its formation. The degradant was identified as the hydroxy methyl derivative (formaldehyde adduct, BMS‐215842) of the drug substance based upon liquid chromatography/mass spectroscopy (LC/MS), liquid chromatography/mass spectroscopy/mass spectroscopy (LC/MS/MS), nuclear magnetic resonance (NMR), and chromatographic comparison to an authentic sample of hydroxymethyl degradation product, BMS‐215842. An assay method for the detection of formaldehyde based on HPLC quantitation of formaldehyde dinitrophenylhydrazone was developed to quantitate its levels in various Polysorbate 80 and PEG 300 excipient lots. A direct relationship between the levels of formaldehyde in the excipients and the formation of the hydroxymethyl degradant was found. To confirm the hypothesis that the formaldehyde impurity in these two excipients contributed to the formation of the hydroxymethyl degradant, several clinical formulation lots were spiked with formaldehyde equivalent to 1, 10, and 100 mg/g of BMS‐204352. A correlation was found between the formaldehyde level and the quantity of the hydroxymethyl degradant formed upon storage at 5 and 25°C. From these experiments, a limit test on the formaldehyde content in polysorbate 80 and PEG 300 can be set as part of a strategy to limit the formation of the degradation product.

Degradation of a Lyophilized Formulation of BMS-204352: Identification of Degradants and Role of Elastomeric Closures

The purpose of this study was to identify two degradation products formed in the parenteral lyophilized formulation of BMS-204352, investigate the possible role of elastomeric closures in their formation, and develop a strategy to minimize/control their formation. The first degradant was identified as the hydroxymethyl derivative (formaldehyde adduct, BMS-215842) of the drug substance formed by the reaction of BMS-204352 with formaldehyde. Structure confirmation was based on liquid chromatography/mass spectroscopy (LC/MS), nuclear magnetic resonance (NMR), and chromatographic comparison to an authentic sample of the hydroxymethyl degradation product, BMS-215842. To confirm the hypothesis that formaldehyde originated from the rubber closure, migrated into the product, and reacted with BMS-204352 drug substance to form the hydroxymethyl degradant, lyophilized drug product was manufactured, the vials were stoppered with two different rubber closure formulations, and its stability was monitored. The formaldehyde adduct degradant was observed only in the drug product vials stoppered with one of the rubber closures that was evaluated. Although formaldehyde has not been detected historically as leachable and is not an added ingredient in the rubber formulation, information obtained from the stopper manufacturer indicated that the reinforcing agent used in the stopper formulation may be a potential source of formaldehyde. The second degradant was identified as the desfluoro hydroxy analog (BMS-188929) based on LC/MS, NMR, and chromatographic comparison to an authentic sample of the desfluoro hydroxy degradation product.

Ethyl methanesulphonate in a parenteral formulation of BMS-214662 mesylate, a selective farnesyltransferase inhibitor: Formation and rate of hydrolysis

The objectives of the present study were to investigate the formation and rate of hydrolysis of ethyl methanesulphonate (EMS) in BMS-214662 mesylate drug substance and parenteral formulation by a gas chromatographic/mass spectrometric (GC/MS) method. EMS levels in the drug substance ranged between 0.3 μg/g and 0.8 μg/g. The parenteral formulation contains ethanol and the reaction between residual free methane sulphonic acid and ethanol may lead to the formation of EMS. Given that EMS is a potent mutagen, it is therefore of vital importance to eliminate or reduce the risk of human exposure. Data indicate no significant increase in the levels of EMS following storage of the drug product for 18 weeks at 25°C or six weeks at 60°C indicating that the potential reaction between ethanol and free methane sulphonic acid may not occur in the BMS-214662 formulation under the storage conditions evaluated and therefore causes no plausible safety concerns of EMS exposure in humans. Kinetic studies were conducted by spiking 200 ppb of EMS into water and the diluted and undiluted parenteral formulation. The rates of hydrolysis of EMS at 25°C followed pseudo-first order kinetics and were determined to be 2.35 × 10−4min−1, 67.4 × 10−4min−1, and 1.32 × 10−4min−1 in water, undiluted, and diluted drug product, respectively.

Impurities in a Lyophilized Formulation of BMS-204352: Identification and Role of Sanitizing Agents

The purpose of this study was to identify two impurities in the parenteral lyophilized formulation of BMS-204352, investigate the role of sanitizing agents as their potential source, evaluate their effect on drug product stability, and develop a strategy to prevent their contamination of the drug product. The two impurities were identified as o-phenylphenol and 4-t-amylphenol based on liquid chromatography/mass spectroscopy (LC/MS) and chromatographic comparison to authentic samples. The LC/MS spectra of commercially available o-phenylphenol and 4-t-amylphenol showed identical patterns of fragmentation and the same retention times as the impurities identified in the BMS-204352 lyophilized product. Levels of these impurities were low and ranged between 0.2–0.3 μg/vial as determined by HPLC and using an authentic external reference standard. To confirm the hypothesis that the commercial sanitizing agents used in the sterile area were the source of these phenolic impurities, several product samples were spiked with the sanitizing agents. Both o-phenylphenol and 4-t-amylphenol were detected in the spiked samples. Further investigation revealed that o-phenylphenol and 4-t-amylphenol are active ingredients of these commercial sanitizing agents. Drug product samples containing the phenolic impurities showed no potency loss following storage at 30, 50, and 70°C indicating these impurities had no adverse effect on product stability. These studies suggest that sanitizing agents used in the sterile area, although may be present at trace levels below typical cleaning procedure detection methods, need to be properly controlled and closely monitored during the manufacturing of injectable products, particularly highly potent drugs. Sanitizing agents, even though not used on product contact surfaces, may potentially contaminate a product through vapor transfer in an open environment.