ABSTRACT
OBJECTIVE: To develop an LC-MS/MS method for the determination of vonoprazan pyroglutamate and vonoprazan fumarate in rat urine to determince the urine excretion of the two drugs in SD rats. METHODS: The detection was performed on an API 4000 tandem mass spectrometer equipped with an electrospray ionization (ESI) source. Multiple reaction monitoring (MRM) was selected with the transitions of m/z 346.2 to 315.2 for TAK-438 P and m/z 237.2 to 194 for IS, respectively. Separation of the analytes was achieved by a Shimadzu liquid chromatography system with an Agelient C18 analytical column (4.6 mm×150 mm, 3.5 μm). Isocratic elution was adopted with mobile phase A (10 mmol•L-1 ammonium acetate and 0.1% formic acid) and mobile phase B (methanol) at the ratio of 40:60, at a flow rate of 0.6 mL•min-1. The total run time was 6 min and the injected sample volume was 5 μL. All the features of the developed method suggested it met the criteria for bioanalytical METHODS recommended by regulatory authorities. The accumulative urine excretion rates of TAK-438 F and TAK-438 P were determined after oral administration of TAK-438 P and equimolar TAK-438 F in SD rats. RESULTS: The accumulative urine excretion rates of the prototype drugs were 2.11% and 2.03%, respectively. The low excretion rates indicated that metabolism might be the major clearance mechanism of TAK-438 P and TAK-438 F. CONCLUSION: This was the first time to establish and validate a simple, rapid and sensitive LC-MS/MS method for the quantification of TAK-438 P. There is no significant difference of the accumulative urine excretion rate between TAK-438 P and TAK-438 F in SD rats, which provides the basis for the druggability of TAK-438 P.
ABSTRACT
Objective:To develop an HPLC method for the determination of vonoprazan pyroglutamate and vonoprazan fumarate. Methods:The column of Intertsil ODS-3 (150 mm×4.6 mm,5 μm) was used. The mobile phase was composed of methanol and a mixture of 0.15% phosphoric acid(ajust pH of 3 with 0.15% triethylamine solution). Gradient elution was adopted and the flow rate was set at 1.0 ml·min-1. The detection wavelength was 230 nm and the column temperature was 30℃. The sample size was 10 μl. Results: The standard curve of vonoprazan pyroglutamate showed a good linearity over the range of 2.060-131.800 μg·ml-1with a correlation coefficient of 0.999 7. The average spiked recovery of vonoprazan pyroglutamate was 99.40% (RSD=0.63%, n=9). The content of three batches of TAK-438 P was 98.7%,99.0% and 98.4%(n=3),respectively. The standard curve of vonoprazan fumarate showed a good linearity over the range of 1.844-118.000 μg·ml-1with a correlation coefficient of 0.999 9. The average spiked recovery of TAK-438 F was 100.67%(RSD =0.52%, n =9). The content of three batches of vonoprazan fumarate was 98.5%,98.2% and 98.9%(n=3),respectively. Conclusion:A reliable and sensitive HPLC method for the quantification of vono-prazan pyroglutamate is established and validated,which provides the basis for the content determination.
ABSTRACT
Objective To develop a practical synthetic process of vonoprazan fumarate with high-yield and lower impurities to meet the quality requirements. Methods By using 5-(2-fluorophenyl)pyrrole-3-carboxaldehyde as the starting material,the qualified vonoprazan fumarate was synthesized via the following steps:①N-sulfonylation and the chloride impurity was removed by recrystalli-zation from MeOH;②the aldehyde was converted to amine by reductive amination,followed by forming the vonoprazan chloride to re-move the dimethylamino impurity;③vonoprazan free base was obtained by neutralization and then converted to fumarate at room tem-perature and finally recrystallized from MeOH/H2O(1:1). Results An impurity controllable synthetic process was developed with a 4％total yield improving. The final product was confirmed by ESI-MS and 1H NMR. Conclusion The synthetic process with single im-purity less than 0.1％and purity above 99.5％was obtained and suitable for scale production of vonoprazan fumarate.