Objective: To establish an ultra-high performance liquid-tandem mass spectrometry (UPLC-MS/MS) detection method for the determination of cetilistat impurity A(2-[[(hexadecyloxy)carbonyl]amino]- 5-methylb enzoic acid) in rat plasma and to apply it in a pharmacokinetic study. Methods: Orlistat was used as the internal standard in rat plasma, and the protein was precipitated by acetonitrile, centrifuged at 10 000 r·min-1 for 10 min,150 μL of supernatant was added to 10 μL of tetrahydrofuran and mixed well, and then sampled for analysis. Using Hypersil GOLD(50 mm×2.1 mm, 1.9 μm) chromatographic column, and the column temperature was 30 ℃, with 0.1% formic acid acetonitrile-0.1% formic acid water (82∶18) as fluidity, the flow rate was 0.5 mL·min-1 . ESI source was used as mass detector in positive ion mode. The precursor→product ion pairs of cetilistat impurity A and olistat were m/z 420.3→160.3 and m/z 496.4→319.3, respectively. Results: The mass concentration of cetilistat impurity A shows a good linear relationship in the concentration range of 0.020 0-10.0 mg·L-1. The REs of stability and accuracy were in the range of 0.1%-6.3%, and the RSDs of precision, recovery, and matrix effect were all below 3.8%. In the pharmacokinetic experiment of three doses of cetilistat impurity A suspension in rats with low(20 mg·kg-1), medium(80 mg·kg-1) and high (320 mg·kg-1) gavage, the peak concentration(Cmax) of cetilistat impurity A was as low as 12.1 mg·L-1 in rat plasma in high dose group. The AUC of the low, medium, and high-dose groups were 22.2, 64.9, and 85.0 mg·L-1·h, respectively.The plasma exposure increased with the dose, but the increase rate was lower than the dose increase rate. The peak time(Tmax) of the three-dose group was between 2.6 and 3.0 h, and the elimination half-life (t1/2z) was between 3.08 and 3.85 h. The three dose groups showed similar absorption and elimination processes. Conclusion: A method for the determination of cetilistat impurity A in rat plasma by ultra-high performance liquid phase-tandem mass spectrometry was established. And the rat plasma exposure is less than the dose-related linear increase in the dose range of 20-320 mg·kg-1 after gavaged with cetilistat-impurity A suspension,and the amount absorbed into blood is relatively small, and the satety problems caused by oral administration are limited.
WANG Xian-zhen
. UPLC-MS/MS method to detect the content of cetilistat impurity A in rat plasma and its application in pharmacokinetics[J]. Chinese Journal of Pharmaceutical Analysis, 2022
, 42(4)
: 637
-642
.
DOI: 10.16155/j.0254-1793.2022.04.10
[1] RAJ P. Cetilistat, a new lipase inhibitor for the treatment of obesity[J].Curr Opin Investig Drugs, 2008, 9(4):414
[2] KOPELMAN P, GROOT GDE H, RISSANEN, et al. Weight loss, hbA1c reduction, and tolerability of cetilistat in a randomized, placebo-controlled phase 2 trial in obese diabetics:comparison with orlistat(Xenical)[J].Obesity(Silver Spring), 2010, 18(1):108
[3] GRAS J. Cetilistat for the treatment of obesity[J].Drugs Today (Barc), 2013,49(12):755
[4] YUAN SF, CHAN JF, CHIK KK, et al. Discovery of the FDA-approved drugs bexarotene,cetilistat, diiodohydroxyquinoline, and abiraterone as potential COVID-19 treatments with a robust two-tier screening system[J].Pharmacol Res, 2020, 159:104960
[5] 阎家麒, 高荣, 谢建平.赛利司他(ATL-962)、利莫那班和赛尼可的研发与市场趋势[J].化工时刊,2008, 22(2):72
YAN JQ, GAO R, XIE JP. Research and development and market trends of anti-obesity such as cetilistat(ATL-962), rimonabant and xenical[J].Chem Ind Times, 2008, 22(2):72
[6] KOPELMAN P, GROOT G, RISSANEN A,et al. Weight loss, HbA1c reduction, and tolerability of cetilistat in a randomized, placebo-controlled phase 2 trial in obese diabetics:comparison with rlistat(Xenical)[J].Obesity(Silver Spring), 2010,18(1):108
[7] YAMADA Y. Cetilistat (ATL-962), a Novel pancreatic lipase inhibitor, ameliorates body weight gain and improves lipid profiles in rats[J].Horm Metab Res, 2008, 40(8):539
[8] BRYSON A, MOTTE S, DUNK C. Reduction of dietary fat absorption by the novel gastrointestinal lipase inhibitor cetilistat in healthy volunteers[J].Br J Clin Pharmacol, 2010, 67(3):309
[9] 沈晓燕.新利司他及其片剂的杂质研究[D].南京:南京工业大学, 2015
SHEN XY. Study on Impurities in Cetilistat and Its Tablets[D].Nanjing:Nanjing University of Technology, 2015
[10] ICH Guideline(Q3A)(R2)Impurities in New Drug Substances[S].2006
[11] 中华人民共和国药典2020年版.四部[S].2020:466
ChP 2020. Vol Ⅳ[S].2020:466
[12] CHEUNG BMY. Satety of antiobesity drugs[J].Ther Adv Drug Saf, 2013, 4(4):171
[13] 张立波,陈蓓,汤谷平. HPLC测定赛利司他有关物质及降解产物[J].中国现代应用药学, 2017,34(3):399
ZHANG LB, CHEN B, TANG GP. Determination of the retated substances of cetilistate by HPLC[J].Chin J Mod Appl Pharm, 2017, 34(3):399
[14] LI XB, MA M, DU CB, et al. Solubility of cetilistat in neat solvents and preferential solvation in (acetone, isopropanol or acetonitrile)+water co-solvent mixtures[J].J Mol Liq, 2017, 242:618
[15] 鲁南制药集团股份有限公司. 一种新利司他中杂质A生物样品的测定方法:中国,201710607602[P].2019-02-01
Lunan Pharmaceutical Group Co., Ltd. A Method for the Determination of Cetilistat Impurity A in Biological Matrix:China, 201710607602[P].2019-02-01
