The study of in vitro synergistic activities of GSK656 with clarithromycin or azithromycin against Mycobacterium abscessus

doi:10.3969/j.issn.1000-6621.2021.02.010

Abstract

Abstract:

Objective The purpose of this study was to assess in vitro drug susceptibility of M.abscessus against the combinations of the 3-aminomethyl 4-halogen benzoxaboroles compound GSK656 with macrolides, including clarithromycin (Clr) and azithromycin (Azm). Methods Eighty nontuberculous mycobacteria isolates were stored the biobank of Beijing Chest Hospital, Capital Medical University between 2016 and 2018, consisting of 30 isolates of M.abscessus, 7 isolates of Mycobacterium kansasii,37 isolates of Mycobacterium intracellulare, 5 isolates of Mycobacterium avium and 1 isolates of Mycobacterium fortuitum. The minimal inhibitory concentrations (MICs) were determined using Microporous Alamar-Blue Assay (MABA). Additionally, the checkerboard titration method was used to determine the synergistic effect between GSK656 and other antimicrobial agents. Results For Clr, the MIC values of M.abscessus isolates ranged from 0.031 mg/L to 16 mg/L. The proportion of Clr resistance was 10.0% (3/30) for M.abscessus. For Azm, the MIC values of M.abscessus isolates ranged from 0.125 to 64 mg/L. The Azm resistance was noted in 30.0% (9/30) of M.abscessus isolates. Additionally, GSK656 also exhibited the potent activity against M.abscessus isolates, the MIC₅₀ and MIC₉₀ of which were 0.25 and 2 mg/L, respectively. In the GSK656-Clr combination, 6 (20.0%,6/30) were synergistic, 23 (76.7%,23/30) were indifferent,1 (3.3%,1/30) was antagonistic. For the GSK656-Azm combination, 3 (10.0%,3/30) was synergistic, 18 (60.0%,18/30) were indifferent, and 9(30.0%,9/30)were antagonistic. Chi-square test results revealed that there was a statistically significant difference in synergistic distribution between GSK-Clr and GSK-Azm group (χ²=8.010,P=0.018). Conclusion Our data demonstrate that GSK656 has promising activity against M.abscessus in vitro. The synergistic distribution between GSK-Clr group and GSK-Azm group is statistically significant. The antagonistic effect is more frequent for GSK-Azm combination than GSK-Clr combination.

Key words: GSK656, Antimicrobial agent, Mycobacteria,atypical, Drug evaluation

GUO Hai-Ping, CHEN Lei, HUO Feng-min, PANG Yu, LI Shan-Shan. The study of in vitro synergistic activities of GSK656 with clarithromycin or azithromycin against Mycobacterium abscessus[J]. Chinese Journal of Antituberculosis, 2021, 43(2): 153-158. doi: 10.3969/j.issn.1000-6621.2021.02.010

Figures/Tables 4

References 22

[1]	McGrath EE, Anderson PB. Increased prevalence of non-tuberculous mycobacteria infection. Lancet, 2007,370(9581):28. doi: 10.1016/S0140-6736(07)61044-7.
[2]	Jing H, Wang H, Wang Y, et al. Prevalence of nontuberculous mycobacteria infection, China, 2004—2009. Emerg Infect Dis, 2012,18(3):527-528. doi: 10.3201/eid1803.110175.
[3]	Hernández-Garduño E, Elwood RK. Increasing incidence of nontuberculous mycobacteria, Taiwan, 2000—2008. Emerg Infect Dis, 2010,16(6):1047-1048. doi: 10.3201/eid1606.100228.
[4]	Lee MR, Sheng WH, Hung CC, et al. Mycobacterium abscessus Complex Infections in Humans. Emerg Infect Dis, 2015,21(9):1638-1646. doi: 10.3201/2109.141634.
[5]	Falkinham JO 3rd. Surrounded by mycobacteria: nontuberculous mycobacteria in the human environment. J Appl Microbiol, 2009,107(2):356-367. doi: 10.1111/j.1365-2672.2009.04161.x.
[6]	Bryant JM, Grogono DM, Greaves D, et al. Whole-genome sequencing to identify transmission of Mycobacterium abscessus between patients with cystic fibrosis: a retrospective cohort study. Lancet, 2013,381(9877):1551-1560. doi: 10.1016/S0140-6736(13)60632-7.
[7]	Bryant JM, Grogono DM, Rodriguez-Rincon D, et al. Emergence and spread of a human-transmissible multidrug-resistant nontuberculous mycobacterium. Science, 2016,354(6313):751-757. doi: 10.1126/science.aaf8156.
[8]	Jeon K, Kwon OJ, Lee NY, et al. Antibiotic treatment of Mycobacterium abscessus lung disease: a retrospective analysis of 65 patients. Am J Respir Crit Care Med, 2009,180(9):896-902. doi: 10.1164/rccm.200905-0704OC.
[9]	Griffith DE, Aksamit T, Brown-Elliott BA, et al. An official ATS/IDSA statement: diagnosis, treatment, and prevention of nontuberculous mycobacterial diseases. Am J Respir Crit Care Med, 2007,175(4):367-416. doi: 10.1164/rccm.200604-571ST.
[10]	Li X, Hernandez V, Rock FL, et al. Discovery of a Potent and Specific M.tuberculosis Leucyl-tRNA Synthetase Inhibitor: (S)-3-(Aminomethyl)-4-chloro-7-(2-hydroxyethoxy)benzo[c][1,2]oxaborol-1(3H)-ol (GSK656). J Med Chem, 2017,60(19):8011-8026. doi: 10.1021/acs.jmedchem.7b00631.
[11]	Rock FL, Mao W, Yaremchuk A, et al. An antifungal agent inhibits an aminoacyl-tRNA synthetase by trapping tRNA in the editing site. Science, 2007,316(5832):1759-1761. doi: 10.1126/science.1142189.
[12]	Dong W, Li S, Wen S, et al. In Vitro Susceptibility Testing of GSK656 against Mycobacterium Species. Antimicrob Agents Chemother, 2020,64(2):e01577-19. doi: 10.1128/AAC.01577-19.
[13]	Zhang Z, Pang Y, Wang Y, et al. Differences in risk factors and drug susceptibility between Mycobacterium avium and Mycobacterium intracellulare lung diseases in China. Int J Antimicrob Agents, 2015,45(5):491-495. doi: 10.1016/j.ijantimicag.2015.01.012.
[14]	赵雁林, 逄宇. 结核病实验室检验规程. 北京: 人民卫生出版社, 2015: 75-87.
[15]	Shen Y, Wang X, Jin J, et al. In Vitro Susceptibility of Mycobacterium abscessus and Mycobacterium fortuitum Isolates to 30 Antibiotics. Biomed Res Int, 2018,2018:4902941. doi: 10.1155/2018/4902941.
[16]	Zhang Z, Lu J, Liu M, et al. In vitro activity of clarithromycin in combination with other antimicrobial agents against Mycobacterium abscessus and Mycobacterium massiliense. Int J Antimicrob Agents, 2017,49(3):383-386. doi: 10.1016/j.ijantimicag.2016.12.003.
[17]	Nessar R, Cambau E, Reyrat JM, et al. Mycobacterium abscessus: a new antibiotic nightmare. J Antimicrob Chemother, 2012,67(4):810-818. doi: 10.1093/jac/dkr578.
[18]	Koh WJ, Jeong BH, Kim SY, et al. Mycobacterial Characteristics and Treatment Outcomes in Mycobacterium abscessus Lung Disease. Clin Infect Dis, 2017,64(3):309-316. doi: 10.1093/cid/ciw724.
[19]	中华医学会结核病学分会. 非结核分枝杆菌病诊断与治疗指南(2020年版). 中华结核和呼吸杂志, 2020,43(11):918-946. doi: 10.3760/cma.j.cn112147-20200508-00570.
[20]	张智健. 脓肿分枝杆菌复合群的药物敏感性分析及其对克拉霉素的耐药机制研究. 北京:中国人民解放军总医院解放军医学院, 2015: 1-222.
[21]	Jesus FP, Ferreiro L, Loreto ÉS, et al. In vitro synergism observed with azithromycin, clarithromycin, minocycline, or tigecycline in association with antifungal agents against Pythium insidiosum. Antimicrob Agents Chemother, 2014,58(9):5621-5625. doi: 10.1128/AAC.02349-14.
[22]	Mukherjee D, Wu ML, Teo JWP, et al. Vancomycin and Clarithromycin Show Synergy against Mycobacterium abscessus In Vitro. Antimicrob Agents Chemother, 2017,61(12):e01298-17. doi: 10.1128/AAC.01298-17.

关系	MIC范围 (mg/L)				FICI范围	菌株数
关系	单药 GSK656	单药 Azm	联合 GSK656	联合 Azm	FICI范围	菌株数
协同	2~8	0.25~1	0.5~2	0.063~0.25	0.375~0.5	3
无关	0.125~4	0.125~64	0.063~2	0.063~32	0.75~2	18
拮抗	0.063~0.5	0.125~1	0.063~1	0.25~4	3~8.5	9

关系	MIC范围 (mg/L)				FICI范围	菌株数
关系	单药 GSK656	单药 Clr	联合 GSK656	联合 Clr	FICI范围	菌株数
协同	0.25~4	0.031~0.125	0.063~1	0.008~1	0.3125~0.5	6
无关	0.063~8	0.031~16	0.063~2	0.008~4	0.625~2	23
拮抗	0.125	0.031	0.125	0.125	5	1

关系	菌株[株(构成比,%)]		χ²值	P值
关系	GSK656-Clr	GSK656-Azm	χ²值	P值
协同	6(20.0)	3(10.0)	8.010	0.018
无关	23(76.7)	18(60.0)
拮抗	1(3.3)	9(30.0)