Differential In Vitro and In Vivo Toxicities of Antimicrobial Peptide Prodrugs for Potential Use in Cystic Fibrosis
Differential In Vitro and In Vivo Toxicities of Antimicrobial Peptide Prodrugs for Potential Use in Cystic Fibrosis
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DOI：
10.1128/aac.00157-16
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发表时间：
2016-05-01
影响因子：
4.9
通讯作者：
Devocelle, Marc
中科院分区：
文献类型：
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作者：
Forde, Eanna;Schuette, Andre;Devocelle, Marc
抗菌肽(AMPs)作为抗菌剂用于治疗囊性纤维化(CF)等多种疾病已引起相当大的兴趣。CF患者肺的挑战性条件需要强大的AMP，这些AMP在高蛋白分解活性的环境中活跃，但也具有低细胞毒性和免疫原性。此前，我们开发了AMP的前药，通过使AMP的抗微生物活性依赖于宿主酶中性粒细胞弹性蛋白酶(NE)来限制AMP治疗的细胞毒作用。然而，细胞毒性仍然是一个问题。在这里，我们描述了进一步优化的AMP前药(PRO-AMP)模型以生产PRO-WMR，与前一组前AMP相比，PRO-WMR的细胞毒性大大降低(对CFBE41-细胞的抑制浓度为50%，>；300muM)。在CF患者富含NE的支气管肺泡灌洗液中，Pro-WMR的杀菌活性增加(从8.4%+/-6.9%增加到91.5%+/-5.8%；P=0.0004)，与以往的Pro-AMP相比，活性差异更大。在小鼠肺部给药的模型中，前AMP修饰降低了宿主毒性，PRO-WMR的毒性低于活性多肽。此前，宿主毒性问题一直阻碍着AMPS的临床应用。然而，开发具有特定用途的AMP，并进行类似于这里描述的将毒性降至最低的修改，可以显著促进其在患者中的潜在使用。这种前药策略与高活性的AMP相结合，有可能为CF患者肺部的挑战性条件产生新的治疗方法。
There has been considerable interest in the use of antimicrobial peptides (AMPs) as antimicrobial agents for the treatment of many conditions, including cystic fibrosis (CF). The challenging conditions of the CF patient lung require robust AMPs that are active in an environment of high proteolytic activity but that also have low cytotoxicity and immunogenicity. Previously, we developed prodrugs of AMPs that limited the cytotoxic effects of AMP treatment by rendering the antimicrobial activity dependent on the host enzyme neutrophil elastase (NE). However, cytotoxicity remained an issue. Here, we describe the further optimization of the AMP prodrug (pro-AMP) model for CF to produce pro-WMR, a peptide with greatly reduced cytotoxicity (50% inhibitory concentration against CFBE41o-cells, > 300 mu M) compared to that of the previous group of pro-AMPs. The bactericidal activity of pro-WMR was increased in NE-rich bronchoalveolar lavage (BAL) fluid from CF patients (range, 8.4% +/- 6.9% alone to 91.5% +/- 5.8% with BAL fluid; P = 0.0004), an activity differential greater than that of previous pro-AMPs. In a murine model of lung delivery, the pro-AMP modification reduced host toxicity, with pro-WMR being less toxic than the active peptide. Previously, host toxicity issues have hampered the clinical application of AMPs. However, the development of application-specific AMPs with modifications that minimize toxicity similar to those described here can significantly advance their potential use in patients. The combination of this prodrug strategy with a highly active AMP has the potential to produce new therapeutics for the challenging conditions of the CF patient lung.