Araujo Marlon Heggdorne de, Muñoz Sánchez Salomé, Simão Thatiana Lopes Biá Ventura, Nowik Natalia, Antunes Stella Schuenck, Pinto Shaft Corrêa, Sorze Davide, Boldrin Francesca, Manganelli Riccardo, Correia Romeiro Nelilma, Lasunskaia Elena B, Verbeek Fons J, Spaink Herman P, Muzitano Michelle Frazão
Laboratório de Produtos Bioativos (LPBio), Instituto de Ciências Farmacêuticas, Universidade Federal do Rio de Janeiro, Campus Macaé, Macaé 27930-560, RJ, Brazil.
Department of Animal Sciences and Health, Institute of Biology (IBL), Leiden University, 2333 BE Leiden, The Netherlands.
Pharmaceuticals (Basel). 2024 Nov 21;17(12):1560. doi: 10.3390/ph17121560.
: Tuberculosis (TB) is one of the leading infectious causes of death worldwide, highlighting the importance of identifying new anti-TB agents. In previous research, our team identified antimycobacterial activity in leaf extract; therefore, this study aims to conduct further exploration of its potential. : Classical chromatography was applied for fractionation and spectrometric techniques were utilized for chemical characterization. For in vitro tests, samples were assessed against and . The toxicity and efficacy of active samples were evaluated in vivo using different zebrafish models. Chemogenomics studies were applied to predict the isolated active compound's potential mode of action. : We performed fractionation of ethanolic extract (EE) and then its dichloromethane fraction (DCM), and the biflavonoid podocarpusflavone A (PCFA) was isolated and identified as a promising active compound. The EE and PCFA were found to be non-toxic to zebrafish larvae and were able to inhibit growth extracellularly. Additionally, PCFA demonstrated antimycobacterial activity within infected macrophages, especially when combined with isoniazid. In addition, the EE, DCM, and PCFA have shown the ability to inhibit growth during in vivo zebrafish larvae yolk infection. Notably, PCFA also effectively countered systemic infection established through the caudal vein, showing a similar inhibitory activity profile to rifampicin, both at 32 µM. A reduction in the transcriptional levels of pro-inflammatory cytokines confirmed the infection resolution. The protein tyrosine phosphatase B (PtpB) of . , which inhibits the macrophage immune response, was predicted as a theoretical target of PCFA. This finding is in agreement with the higher activity observed for PCFA intracellularly and in vivo on zebrafish, compared with the direct action in . : Here, we describe the discovery of PCFA as an intracellular inhibitor of and provide evidence of its in vivo efficacy and safety, encouraging its further development as a combination drug in novel therapeutic regimens for TB.
结核病（TB）是全球主要的感染性致死病因之一，凸显了识别新型抗结核药物的重要性。在先前的研究中，我们团队在树叶提取物中发现了抗分枝杆菌活性；因此，本研究旨在对其潜力进行进一步探索。采用经典色谱法进行分离，并利用光谱技术进行化学表征。对于体外试验，针对[具体菌株1]和[具体菌株2]对样品进行评估。使用不同的斑马鱼模型在体内评估活性样品的毒性和疗效。应用化学基因组学研究来预测分离出的活性化合物的潜在作用模式。我们对[植物名称]乙醇提取物（EE）进行了分离，然后对其二氯甲烷馏分（DCM）进行分离，分离并鉴定出双黄酮罗汉松黄酮A（PCFA）为一种有前景的活性化合物。发现EE和PCFA对斑马鱼幼虫无毒，并且能够在细胞外抑制[具体菌株]的生长。此外，PCFA在受感染的巨噬细胞内表现出抗分枝杆菌活性，尤其是与异烟肼联合使用时。此外，EE、DCM和PCFA在斑马鱼幼虫卵黄感染的体内实验中显示出抑制[具体菌株]生长的能力。值得注意的是，PCFA在32μM时也能有效对抗通过尾静脉建立的全身感染，显示出与利福平相似的抑制活性谱。促炎细胞因子转录水平的降低证实了感染得到缓解。预测[结核分枝杆菌名称]的蛋白酪氨酸磷酸酶B（PtpB），其抑制巨噬细胞免疫反应，是PCFA的理论靶点。这一发现与在细胞内和斑马鱼体内观察到的PCFA比在[体外实验环境]中的直接作用具有更高活性相一致。在此，我们描述了PCFA作为[结核分枝杆菌名称]细胞内抑制剂的发现，并提供了其体内疗效和安全性的证据，鼓励将其作为新型结核病治疗方案中的联合药物进一步开发。
