Objective: To establish a rapid and accurate method of UPLC-MS/MS for the determination of macozinone(PBTZ 169) concentration, a novel active compound against multi-resistant tuberculosis, in rat plasma and to study its pharmacokinetics in rats. Methods: Carbamazepine was used as internal standard. After the protein was precipitated by acetonitrile, the plasma samples were separated on the XBridge BEH C18(100 mm×2.1 mm, 2.5 μm) chromatographic column through gradient elution by acetonitrile and 0.1% formic acid as mobile phase. The flow rate was 0.5 mL·min-1 and sample volume was 10 μL. Then the analytes were detected by the electrospray ionization (ESI) sourcein with positive ion scanning. The detection conditions of PBTZ 169 and internal standard(carbamazepine) by mass spectrometry were m/z 457.2→344.1 and m/z 237.2→194.2, respectively. After the rats were given 20 mg·kg-1 PBTZ 169 and the plasma concentration of PBTZ 169 was detected at different time points. The pharmacokinetic parameters were calculated and statistically analyzed. Results: The mass concentration of PBTZ 169 had a good linear relationship between 0.7 and 740.0 ng·mL-1(r=0.996 0). The extraction recovery of PBTZ 169 and internal standard were between 99.5% and 110.2% and the RSDs of intra day and inter day were ranged from 1.3% to 4.2%. PBTZ 169 plasma samples were placed at room temperature for 6 h, the automatic sampler was placed for 24 h after the blood sample was processed, refrigerated at -70 ℃ for 15 d and repeatedly freeze-thaw at -70 ℃ for 3 times. Finally, the RSDs were all lower than 15%. After oral intragastric administration of PBTZ 169 to rats, the main pharmacokinetic parameters of PBTZ 169 in rats were as follows: Cmax was (374.8±116.3) ng·mL-1, Tmax was (0.8±0.1) h, AUC0~∞ was (1 180.0±383.5) ng·h·mL-1, T1/2 was (3.7±0.7) h, MRT0~∞ was (4.7±1.2) h. Conclusion: This method is simple, rapid, sensitive and suitable for pharmacokinetic study of PBTZ 169.
ZHI Xu-ran, XING Xiao-qing, LI Ying, LIU Hong-tao
. Determination of the active compound against multi-resistant tuberculosis macozinone (PBTZ 169) concentration in rat plasma by UPLC-MS/MS method and its pharmacokinetics*[J]. Chinese Journal of Pharmaceutical Analysis, 2023
, 43(1)
: 126
-132
.
DOI: 10.16155/j.0254-1793.2023.01.15
[1] CARDONA PJ. Pathogenesis of tuberculosis and other mycobacteriosis[J].Enferm Infecc Microbiol Clin(Engl Ed), 2018, 36(1):38
[2] SCHITO M, MIGLIORI GB, FLETCHER HA, et al. Perspectives on advances in tuberculosis diagnostics, drugs, and vaccines[J].Clin Infect Dis, 2015, 61(Suppl 3):S102
[3] World Health Organization. Global Tuberculosis Report 2021[R].Geneva: WHO, 2021
[4] 雷倩, 赵嫄, 王皓, 等. 3种二线抗结核药物血药浓度测定及其影响因素[J].药物分析杂志,2020, 40(8):1405
LEI Q, ZHAO Y, WANG H, et al. Determination of plasma concentration of three second-line anti-tuberculous drugs and their influencing factors[J].Chin J Pharm Anal, 2020, 40(8):1405
[5] PONTALI E, VISCA D, CENTIS R, et al. Multi and extensively drug-resistant pulmonary tuberculosis: advances in diagnosis and management[J].Curr Opin Pulm Med, 2018, 24(3):244
[6] TREFZER C, RENGIFO-GONZALEZ M, HINNER MJ, et al. Benzothiazinones: prodrugs that covalently modify the decaprenylphosphoryl-β-D-ribose 2'-epimerase DprE1 of mycobacterium tuberculosis[J].J Am Chem Soc, 2010, 132(39):13663
[7] PASCA MR, DEGIACOMI G, RIBEIRO A, et al. Clinical isolates of mycobacterium tuberculosis in four european hospitals are uniformly susceptible to benzothiazinones[J].Antimicrob Agents Chemother, 2010, 54(4):1616
[8] LECHARTIER B, HARTKOORN RC, COLE ST. In vitro combination studies of benzothiazinone lead compound BTZ043 against mycobacterium tuberculosis[J].Antimicrob Agents Chemother, 2012, 56(11):5790
[9] LECHARTIER B, COLE ST. Mode of action of clofazimine and combination therapy with benzothiazinones against mycobacterium tuberculosis[J].Antimicrob Agents Chemother, 2015, 59(8):4457
[10] STEPANOVA ES, BARSEGYAN SS, MAKARENKOVA LM, et al. Development and validation of a quantitative determination method for the antitubercular drug PBTZ169 in biological media[J].Pharm Chem J, 2018, 52(3):248
[11] LUPIEN A, VOCAT A, FOO CS, et al. An optimized background regimen for treatment of active tuberculosis with the next-generation benzothiazinone macozinone (PBTZ169)[J].Antimicrob Agents Chemother, 2018, 62(11):840
[12] MAKAROV V, LECHARIER B, ZHANG M, et al. Towards a new combination therapy for tuberculosis with next generation benzothiazinones[J].EMBO Mol Med, 2014, 6(3):372
[13] ZHANG G, ALDRICH CC. Macozinone: revised synthesis and crystal structure of a promising new drug for treating drug-sensitive and drug-resistant tuberculosis[J].Acta Crystallogr C Struct Chem, 2019, 75(8):1031
[14] SPAGGIARI D, DESFONTAINE V, CRUCHON S, et al. Development and validation of a multiplex UHPLC-MS/MS method for the determination of the investigational antibiotic against multi-resistant tuberculosis macozinone (PBTZ169) and five active metabolites in human plasma[J].PloS One, 2019, 14(5):e0217139
[15] WANG A, LV K, TAO Z, et al. Identification of benzothiazinones containing an oxime functional moiety as new anti-tuberculosis agents[J].Eur J Med Chem, 2019, 181: 111595
