Development of LC/MS/MS assay for the determination of 5-ethyl-2-{5-[4-(2-hydroxyethyl)piperazine-1-sulfonyl]-2-propoxyphenyl}-7-propyl-3,5-dihydropyrrolo[3,2-d]pyrimidin-4-one (SK3530) in human plasma: application to a clinical pharmacokinetic study.
5-Ethyl-2-{5-[4-(2-hydroxyethyl)piperazine-1-sulfonyl]-2-propoxyphenyl}-7-propyl-3,5-dihydropyrrolo[3,2-d]pyrimidin-4-one (SK3530) is a new phosphodiesterase type-5 inhibitor currently undergoing a Phase III investigation for the treatment of male erectile dysfunction. This study first describes a rapid and sensitive LC/MS/MS assay method for the quantification of SK3530 and its major metabolite, SK3541, in human plasma. The assay was validated to demonstrate the specificity, linearity, recovery, lower limit of quantification (LLOQ), accuracy, and precision. The multiple reaction monitoring was based on the transition of m/z=532.5-->99.1 for SK3530, 488.6-->295.5 for SK3541, and 520.3-->99.1 for SK3304 (internal standard). The assay utilized a single liquid-liquid extraction and isocratic elution, and the LLOQ was 1 ng/ml using 0.2 ml human plasma. The assay was linear over a range from 1 to 1000 ng/ml for both SK3530 and SK3541, with correlation coefficients >0.9999. The mean intra- and inter-day assay accuracy ranged from 94.7 to 101.6% and 96.8 to 101.1% for SK3530 and 92.6-105.7% and 97.4-107.8% for SK3541, respectively. The mean intra- and inter-day precision was between 7.2-12.1% and 5.7-7.4% for SK3530 and 4.6-13.2% and 5.0-14.1% for SK3541, respectively. The developed assay was applied to a clinical pharmacokinetic study after oral administration of SK3530 in healthy male volunteers (dose 100 mg).
Shin
Beom Soo
BS
College of Pharmacy, Catholic University of Daegu, Gyeongsan-si, Gyeongbuk, Republic of Korea.
Hu
Seul Ki
SK
College of Pharmacy, Sungkyunkwan University, Suwon, Kyeonggi-do, Republic of Korea.
Kim
John
J
College of Pharmacy, Sungkyunkwan University, Suwon, Kyeonggi-do, Republic of Korea.
Oh
Joon Gyo
JG
Life Science R&D Center, SK Chemicals Ltd., Suwon, Kyeonggi-do, Republic of Korea.
Youn
Won-No
WN
Life Science R&D Center, SK Chemicals Ltd., Suwon, Kyeonggi-do, Republic of Korea.
Lee
Bongyong
B
Life Science R&D Center, SK Chemicals Ltd., Suwon, Kyeonggi-do, Republic of Korea.
Um
Key An
KA
Life Science R&D Center, SK Chemicals Ltd., Suwon, Kyeonggi-do, Republic of Korea.
Kim
Dae-Kee
DK
College of Pharmacy, Ewha Womans University, Seoul, Republic of Korea.
Lee
Ju Young
JY
R&D Center, In2Gen Co., Ltd., Seoul, Republic of Korea.
Yoo
Sun Dong
SD
College of Pharmacy, Sungkyunkwan University, Suwon, Kyeonggi-do, Republic of Korea. Electronic address: sdyoo@skku.ac.kr.
D000284:Administration, Oral; D000328:Adult; D002138:Calibration; D002851:Chromatography, High Pressure Liquid; D007172:Erectile Dysfunction / Q000188:drug therapy; D006801:Humans; D008297:Male; D015394:Molecular Structure; D010601:Pharmacology, Clinical / Q000379:methods*; D011744:Pyrimidinones / Q000097:blood* / Q000493:pharmacokinetics* / Q000627:therapeutic use; D011786:Quality Control; D012015:Reference Standards; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity; D021241:Spectrometry, Mass, Electrospray Ionization; D013450:Sulfones / Q000097:blood* / Q000493:pharmacokinetics* / Q000627:therapeutic use; D053719:Tandem Mass Spectrometry
C518762:2-(5-(4-(2-hydroxyethyl)piperazin-1-ylsulfonyl)-2-n-propoxyphenyl)-5-ethyl-7-n-propyl-3,5-dihydro-4H-pyrrolo(3,2-d)pyrimidin-4-one dihydrochloride; D011744:Pyrimidinones; D013450:Sulfones