Study on the metabolic interaction mechanism between the novel compound WX-081 and clarithromycin

doi:10.19982/j.issn.1000-6621.20240483

Abstract

Abstract:

Objective: To explore the effect of the new drug sudapyridine (WX-081) on liver enzymes and its interaction with clarithromycin. Methods: Take rat liver microsomes/human liver microsomes and add a mixture of cytochrome P450 (CYP450) probe substrates to the working solution. Add sudapyridine and clarithromycin working solutions of different concentrations, and take the supernatant for liquid chromatography mass spectrometry (LC-MS/MS) analysis. Calculate the concentration of metabolites in the system using the standard curve method, calculate the amount of metabolites generated, and calculate the remaining activity based on the changes in the solvent control group. Results: Under the conditions of human liver microsomal assay, sudapyridine showed weak inhibitory effects on CYP1A2, CYP2B6, CYP2C8, and CYP3A4/5 at concentrations of 0.1, 0.5, 1.5, 5, 10, 25, 50, and 100 μmol/L, and moderate or strong inhibitory effects on CYP2C9, CYP2C19, and CYP2D6; The inhibitory concentration 50% (IC₅₀) of CYP2C9 was 5.533 μmol/L, CYP2C19 was <0.1 μmol/L, and CYP2D6 was 15.59 μmol/L. Under the conditions of rat liver microsomal assay, sudapyridine showed weak inhibitory effects on CYP1A2, CYP2B6, CYP2C9, and CYP3A4/5 at concentrations of 0.1, 0.5, 1.5, 5, 10, 25, 50, and 100 μmol/L, and moderate inhibitory effects on CYP2C8, CYP2C19, and CYP2D6; The IC₅₀ of CYP2C8 was 49.70 μmol/L, CYP2C19 was 10.06 μmol/L, and CYP2D6 was 17.55 μmol/L. In the human liver microsomal assay, clarithromycin showed inhibitory effects on sudapyridine at concentrations of 1, 2, 5, 10, 50, 100, 150, and 200 μmol/L, with an IC₅₀ of 22.74 μmol/L; In the rat liver microsomal assay, clarithromycin showed a slight inhibitory effect on sudapyridine at concentrations of 1, 2, 5, 10, 50, 100, 150, and 200 μmol/L, with an IC₅₀ of 85.61 μmol/L. Conclusion: Sudapyridine has a moderate inhibitory effect on CYP2C9 and CYP2D6 in human liver microsomes; sudapyridine has a moderate inhibitory effect on CYP2C19 and CYP2D6 in rat liver microsomes. In human liver microsomes, clarithromycin has a strong inhibitory effect on sudapyridine.

Key words: Mycobacterium, Clarithromycin, WX-081, Molecular mechanisms of pharmacological action

CLC Number:

R969.2

Li Qi, Wang Yujin, Wang Xueyu, Chu Naihui, Nie Wenjuan. Study on the metabolic interaction mechanism between the novel compound WX-081 and clarithromycin[J]. Chinese Journal of Antituberculosis, 2025, 47(2): 142-149. doi: 10.19982/j.issn.1000-6621.20240483

Figures/Tables 5

CYP酶亚型	舒达吡啶促变药			CYP酶亚型	舒达吡啶促变药
CYP酶亚型	浓度 (μmol/L)	剩余活性 (IU, $x ˉ$ ±s)	IC₅₀ (μmol/L)	CYP酶亚型	浓度 (μmol/L)	剩余活性 (IU, $x ˉ$ ±s)	IC₅₀ (μmol/L)
1A2			>100	2B6			>100
	0	100.0±7.4			0	100.0±4.6
	0.1	82.2±5.3			0.1	82.2±8.7
	0.5	84.8±4.6			0.5	82.2±1.1
	1.5	75.1±5.7			1.5	86.7±9.1
	5	78.5±0.6			5	81.9±6.2
	10	78.6±3.0			10	83.6±3.2
	25	69.6±5.4			25	82.6±3.7
	50	69.9±5.3			50	82.9±0.8
	100	68.7±11.8			100	85.9±47.0
2C8			>100	2C9			5.533
	0	100.0±5.1			0	100.0±3.0
	0.1	97.4±4.1			0.1	64.1±6.0
	0.5	100.9±4.0			0.5	61.2±7.9
	1.5	93.9±5.1			1.5	60.9±10.5
	5	93.4±2.0			5	51.2±2.8
	10	96.2±9.9			10	58.2±0.3
	25	81.5±8.0			25	50.9±6.5
	50	71.8±5.0			50	50.4±1.7
	100	60.7±3.1			100	48.8±2.9
2C19			<0.1	2D6			15.59
	0	100.0±5.7			0	100.0±3.8
	0.1	49.6±12.2			0.1	49.6±4.6
	0.5	66.4±5.0			0.5	57.2±6.0
	1.5	未测出			1.5	45.7±2.9
	5	51.0±15.6			5	47.9±2.7
	10	52.1±15.8			10	47.7±4.0
	25	36.6±14.8			25	37.3±2.9
	50	45.0±13.1			50	40.0±1.2
	100	28.2±1.8			100	35.6±3.7
3A4/5			>100	3A4/5			>100
	0	100.0±3.4			0	100.0±1.4
	0.1	56.2±17.5			0.1	91.8±7.1
	0.5	76.1±9.6			0.5	103.6±3.8
	1.5	62.1±9.2			1.5	96.1±2.8
	5	63.6±2.6			5	93.7±4.9
	10	69.0±3.3			10	97.9±1.7
	25	49.9±3.7			25	87.5±1.6
	50	60.7±8.3			50	95.2±10.6
	100	53.4±3.4			100	92.5±3.2

CYP酶亚型	舒达吡啶促变药			CYP酶亚型	舒达吡啶促变药
CYP酶亚型	浓度 (μmol/L)	剩余活性 (IU, $x ˉ$ ±s)	IC₅₀ (μmol/L)	CYP酶亚型	浓度 (μmol/L)	剩余活性 (IU, $x ˉ$ ±s)	IC₅₀ (μmol/L)
1A2			>100	2B6			>100
	0	100.0±4.2			0	100.0±3.8
	0.1	105.3±3.4			0.1	98.2±7.3
	0.5	97.0±8.1			0.5	93.8±2.5
	1.5	108.1±5.9			1.5	92.8±1.8
	5	107.4±3.1			5	85.5±4.5
	10	105.7±8.0			10	87.9±2.0
	25	97.4±4.6			25	84.7±5.2
	50	90.7±7.8			50	79.3±2.5
	100	83.9±2.7			100	80.3±4.9
2C8			49.70	2C9			>100
	0	100.0±23.8			0	100.0±11.3
	0.1	63.2±17.9			0.1	84.8±7.6
	0.5	64.8±14.0			0.5	93.1±3.4
	1.5	80.4±15.0			1.5	83.1±8.3
	5	56.9±13.3			5	87.8±5.5
	10	69.2±17.4			10	80.2±4.9
	25	44.5±7.5			25	78.3±14.7
	50	48.8±8.7			50	73.2±4.7
	100	24.7±8.4			100	77.1±6.8
2C19			10.06	2D6			17.55
	0	100.0±1.1			0	100.0±5.0
	0.1	71.8±0.2			0.1	88.0±2.2
	0.5	未测出			0.5	70.0±2.9
	1.5	67.2±3.1			1.5	73.6±3.2
	5	61.6±2.5			5	63.5±1.2
	10	57.5±3.7			10	49.4±1.6
	25	52.6±2.7			25	36.5±1.7
	50	49.7±4.6			50	29.9±6.0
	100	46.8±2.1			100	24.0±2.7
3A4/5			>100	3A4/5			>100
	0	100.0±4.4			0	100.0±4.3
	0.1	121.4±13.5			0.1	100.3±6.0
	0.5	109.8±6.6			0.5	90.1±0.8
	1.5	121.5±5.0			1.5	95.2±2.0
	5	126.5±7.9			5	90.5±3.6
	10	117.2±6.6			10	91.7±1.8
	25	114.5±16.7			25	87.9±2.5
	50	114.5±12.5			50	88.2±4.4
	100	95.1±4.3			100	84.4±3.0

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CYP酶亚型	底物	Km (mol/L)	最终体系底物浓度(μmol/L)	底物工作液浓度(μmol/L)	被检测物
1A2	非那西丁	10~50	50	2500	对乙酰氨基酚
2B6	依法韦仑	17~23	20	1000	8-羟基依法韦仑
2C8	紫杉醇	5.4~19	20	1000	6-羟基紫杉醇
2C9	甲苯磺丁脲	100~200	140	7000	4-羟基甲苯磺丁脲
2C19	奥美拉唑	17~26	20	1000	5-羟基奥美拉唑
2D6	右美沙芬	5	7	350	右羟吗喃
3A4/5	睾酮	50~100	75	3750	6β-羟基睾酮
3A4/5	咪达唑仑	2.5~5	5	250	1-羟基咪达唑仑

微粒体种类	克拉霉素受试药物			微粒体种类	克拉霉素受试药物
微粒体种类	浓度 (μmol/L)	剩余活性 (IU,$\bar{x}±s$)	IC₅₀ (μmol/L)	微粒体种类	浓度 (μmol/L)	剩余活性 (IU,$\bar{x}±s$)	IC₅₀ (μmol/L)
人肝微粒体			22.74	大鼠肝微粒体			85.61
	0	100.0±2.7			0	100.0±21.0
	1	98.6±2.0			1	89.8±15.6
	2	103.1±8.4			2	73.6±4.2
	5	91.1±6.7			5	98.6±8.1
	10	72.2±2.0			10	96.9±12.3
	50	39.2±0.6			50	62.3±6.4
	100	23.4±0.4			100	62.8±9.8
	150	19.0±1.7			150	37.2±1.9
	200	15.4±0.8			200	38.7±22.0

基质	被检测物	标准曲线	线性范围(ng/ml)	相关系数(r)
人肝微粒体	舒达吡啶M3工作液	y=1.770445×10^-4x+2.122334×10^-5	1~1000	0.997
大鼠肝微粒体	舒达吡啶M3工作液	y=2.236211×10^-5x+1.753353×10^-5	1~1000	0.997