Preliminary study on pharmacokinetics/pharmacodynamics of five new anti-tuberculosis drugs in mice

doi:10.3969/j.issn.1000-6621.2021.10.015

Abstract

Abstract:

Objective To evaluate the pharmacokinetic/pharmacodynamics (PK/PD) of five anti-tuberculosis drugs including pretomanid (PA-824), delamanid (Dlm), clofazimine (Cfz), bedaquiline (Bdq) and PBTZ-169 in BALB/c mice tuberculosis model at different doses. Methods BALB/c mice were infected with Mycobacterium tuberculosis (H37Rv) by aerosol. Five drugs were divided into high, middle and low dose groups. After 4 and 8 weeks of treatment, the mice were dissected and the colony forming unit (CFU) of spleens and lungs were counted. At the same time, the lungs and spleens homogenate were inoculated on the plate containing 4×minimum inhibitory concentration (MIC) to observe the colony growth. Three hours after the last dose was given to each group, the orbital blood was collected and the drug concentration were determined by HPLC-MS/MS. Results The lowest effective dose of each drug was PA-824 30 mg/kg, Dlm 5 mg/kg, Cfz 5 mg/kg, Bdq 6.25 mg/kg, PBTZ-169 20 mg/kg, which obtained according to the significant difference of lung CFU count between treatment and control groups. From the lowest effective dose, the CFU counts in the lungs of mice were decreased in all treatment groups. At the high-dose of Bdq and PA-824, the CFU counts in the lungs of mice decreased by 2.99 and 2.66 log₁₀ values respectively. At 8 weeks, the plasma concentrations (C_3h) of each drug at the lowest effective dose were PA-824 (7.60±1.28) μg/ml, Dlm (0.20±0.05) μg/ml, Cfz (0.26±0.02) μg/ml, Bdq (0.22±0.07) μg/ml, PBTZ-169 (74.56±17.80) ng/ml, respectively. At high dose, when C_max/MIC was higher than the specific value (PA-824 100 mg/kg, C_max/MIC>147; Dlm 10 mg/kg, C_max/MIC>100; Cfz 20 mg/kg, C_max/MIC>2; Bdq 25 mg/kg, C_max/MIC>13.6; PBTZ-169 20 mg/kg, C_max/MIC>150), drug tolerance/phenotypic resistance was not easy to occur. Conclusion At the effective dose of the five anti-tuberculosis durgs, the plasma concentration increased with the increase of the dosage, but the CFU count was negatively correlated with the dosage. Our study showed that C_max/MIC was a good PK/PD parameter correlation with treatment effect, which could provide guidance for the clinical use of these drugs.

Key words: Tuberculosis, Pharmacokinetics, Drug resistance, Interdisciplinary studies

ZHU Hui, FU Lei, ZHANG Wei-yan, WANG Bin, CHEN Xi, LU Yu. Preliminary study on pharmacokinetics/pharmacodynamics of five new anti-tuberculosis drugs in mice[J]. Chinese Journal of Antituberculosis, 2021, 43(10): 1056-1065. doi: 10.3969/j.issn.1000-6621.2021.10.015

Figures/Tables 11

References 19

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组别	剂量(mg/kg)	4周(g,$\bar{x}\pm s$)	t值	P值	8周(g,$\bar{x}\pm s$)	t值	P值
对照组		21.44±1.07			21.86±0.85
PA-824
L组	10	20.87±0.98	-0.883	0.403	22.56±1.05	1.169	0.276
M组	30	21.97±0.57	0.972	0.359	23.04±0.81	2.261	0.054
H组	100	22.49±0.71	1.829	0.105	22.35±1.01	0.838	0.426
Dlm
L组	2.5	21.56±0.37	0.241	0.816	22.56±0.85	1.306	0.228
M组	5	21.63±1.33	0.244	0.814	23.30±1.06	2.389	0.044
H组	10	21.39±1.05	-0.078	0.940	21.51±1.03	-0.577	0.580
Cfz
L组	5	22.01±1.04	0.846	0.422	23.20±1.20	2.047	0.075
M组	10	21.08±1.82	-0.385	0.710	22.41±1.18	0.854	0.418
H组	20	20.76±0.79	-1.152	0.283	22.82±0.67	1.988	0.082
Bdq
L组	6.25	22.14±1.12	1.016	0.339	22.10±0.48	0.556	0.593
M组	12.5	21.68±0.66	0.417	0.688	23.52±0.35	4.064	0.004
H组	25	21.32±1.05	-0.188	0.855	22.78±0.66	1.923	0.091
PBTZ-169
L组	5	21.30±0.48	-0.275	0.790	22.45±0.88	1.095	0.305
M组	10	21.37±0.38	-0.138	0.894	23.66±0.69	3.690	0.006
H组	20	20.84±0.75	-1.032	0.332	23.45±0.41	3.798	0.005

组别	剂量(mg/kg)	4周时浓度(μg/ml,$\bar{x}\pm s$)	8周时浓度(μg/ml,$\bar{x}\pm s$)	参考MIC^a(μg/ml)
PA-824				0.07
L组	10	3.75±0.92	2.84±0.61
M组	30	9.05±1.70	7.60±1.28
H组	100	17.95±2.87	14.68±1.02
Dlm				0.004
L组	2.5	0.02±0.01	0.20±0.06
M组	5	0.24±0.12	0.20±0.05
H组	10	0.43±0.17	0.40±0.07
Cfz				0.24
L组	5	0.22±0.01	0.26±0.02
M组	10	0.23±0.04	0.43±0.05
H组	20	0.36±0.08	0.47±0.07
Bdq				0.022~0.05
L组	6.25	0.31±0.10	0.22±0.07
M组	12.5	0.32±0.14	0.42±0.07
H组	25	0.68±0.25	0.68±0.26
PBTZ-169				0.4~0.5^b
L组	5	7.25±1.69^b	6.91±1.52^b
M组	10	18.70±6.69^b	21.23±4.45^b
H组	20	51.54±20.35^b	74.56±17.80^b

组别	剂量(mg/kg)		肺(log₁₀CFU,$\bar{x}\pm s$)		t值^a		P值^a		脾(log₁₀CFU,$\bar{x}\pm s$)		t值^a		P值^a
对照组
治疗前14d			4.70±0.11						1.48±0.00
治疗当天			5.21±0.08						3.60±0.18
治疗8周			4.90±0.16						3.08±0.44
PA-824
L组	10		4.72±0.19		-1.542		0.162		4/5为0,1只为2.08		-5.775		<0.001
M组	30		4.00±0.17		-8.654		<0.001		0		-15.475		<0.001
H组	100		2.24±0.22		-21.951		<0.001		0		-15.475		<0.001
Dlm
L组	2.5		4.75±0.20		-1.255		0.245		4/5为0,1只为2.08		-5.775		<0.001
M组	5		4.55±0.14		-3.627		0.007		4/5为0,1只为1.48		-7.803		<0.001
H组	10		3.54±0.16		-13.574		<0.001		0		-15.475		<0.001
Cfz
L组	5		4.07±0.12		-9.405		<0.001		2.47±0.19		-2.799		0.023
M组	10		4.02±0.27		-6.231		<0.001		2.40±0.80		-1.657		0.136
H组	20		3.41±0.28		-10.397		<0.001		2.06±0.23		-4.538		0.002
Bdq
L组		6.25		4.61±0.15		-2.918		0.019		2.90±0.33		-0.731	0.485
M组		12.5		3.85±0.51		-4.405		0.008		4/5为0,1只为2.38		-5.045	0.001
H组		25		1.91±1.08		-6.109		<0.001		0		-15.475	<0.001
PBTZ-169
L组		5		5.54±0.22		5.306		0.001		3.16±0.17		0.381	0.713
M组		10		4.71±0.23		-1.505		0.171		0		-15.475	<0.001
H组		20		3.36±0.21		-12.999		<0.001		4/5为0,1只为1.48		-7.803	<0.001

培养板脏器	L组(10mg/kg)		M组(30mg/kg)		H组(100mg/kg)
培养板脏器	不含药培养板	含药培养板	不含药培养板	含药培养板	不含药培养板	含药培养板
不含活性炭培养板
肺脏	+++	1只为 1CFU	74~171CFU	未生长	3~10CFU	未生长
脾脏	4只未生长, 1只为 4CFU	未生长	未生长	未生长	未生长	未生长
活性炭含药培养板
肺脏	-	100~300CFU	-	44~150CFU	-	0~73CFU
脾脏	-	4只未生长, 1只为 4CFU	-	未生长	-	未生长

培养板脏器	L组(2.5mg/kg)		M组(5mg/kg)		H组(10mg/kg)
培养板脏器	不含药培养板	含药培养板	不含药培养板	含药培养板	不含药培养板	含药培养板
不含活性炭培养板
肺脏	+++	未生长	+++	未生长	85~210CFU	未生长
脾脏	4只未生长, 1只为 4CFU	未生长	4只未生长, 1只为 1CFU	未生长	未生长	未生长
活性炭含药培养板
肺脏	-	1~200CFU	-	3~150CFU	-	1~16CFU
脾脏	-	4只未生长, 1只为 1CFU	-	4只未生长, 1只为1CFU	-	未生长