Abstract:
The increasing prevalence of antimicrobial-resistant Staphylococcus aureus strains, especially methicillin-resistant S. aureus (MRSA), poses a threat to successful antibiotic treatment. Unsuccessful attempts to develop a vaccine and rising resistance to last-resort antibiotics urge the need for alternative treatments. Host-directed therapy (HDT) targeting critical intracellular stages of S. aureus emerges as a promising alternative, potentially acting synergistically with antibiotics and reducing the risk of de novo drug resistance. We assessed 201 ATP-competitive kinase inhibitors from Published Kinase Inhibitor Sets (PKIS1 and PKIS2) against intracellular MRSA. Seventeen hit compounds were identified, of which the two most effective and well-tolerated hit compounds (i.e., GW633459A and GW296115X) were selected for further analysis. The compounds did not affect planktonic bacterial cultures, while they were active in a range of human cell lines of cervical, skin, lung, breast and monocyte origin, confirming their host-directed mechanisms. GW633459A, structurally related to lapatinib, exhibited an HDT effect on intracellular MRSA independently of its known human epidermal growth factor receptor (EGFR)/(HER) kinase family targets. GW296115X activated adenosine monophosphate-activated protein kinase (AMPK), thereby enhancing bacterial degradation via autophagy. Finally, GW296115X not only reduced MRSA growth in human cells but also improved the survival rates of MRSA-infected zebrafish embryos, highlighting its potential as HDT.
摘要:
抗菌耐药金黄色葡萄球菌菌株的流行,尤其是耐甲氧西林金黄色葡萄球菌(MRSA),对成功的抗生素治疗构成威胁。开发疫苗的尝试不成功,对最后手段抗生素的耐药性上升,这促使人们需要替代疗法。针对金黄色葡萄球菌关键细胞内阶段的宿主导向疗法(HDT)成为一种有希望的替代方案,可能与抗生素协同作用,并降低从头耐药的风险。我们评估了来自已发布的激酶抑制剂集(PKIS1和PKIS2)的201种ATP竞争性激酶抑制剂对细胞内MRSA的影响。确定了17个被击中的化合物,其中两种最有效和耐受性最好的化合物(即,选择GW633459A和GW296115X)进行进一步分析。这些化合物不会影响浮游细菌培养,虽然它们在一系列人类宫颈细胞系中活跃,皮肤,肺,乳腺和单核细胞起源,确认他们的主机导向机制。GW633459A,在结构上与拉帕替尼有关,表现出对细胞内MRSA的HDT效应,与其已知的人表皮生长因子受体(EGFR)/(HER)激酶家族靶标无关。GW296115X激活一磷酸腺苷激活蛋白激酶(AMPK),从而通过自噬增强细菌降解。最后,GW296115X不仅降低了MRSA在人类细胞中的生长,而且提高了MRSA感染斑马鱼胚胎的存活率,突出其作为HDT的潜力。