Andrade Beatriz A, Dos Santos Augusto L, Ferreira Dayana A S, Abiuzi Mariana B, Vieira Daniel P, Gonçalves Marina M, Lago João Henrique G, Sartorelli Patricia, Tempone Andre G
Pathophysiology Laboratory, Instituto Butantan, Av. Vital Brazil, 1500, São Paulo, SP 05503-900, Brazil.
Postgraduate Program in Infectious Diseases and Global Health - Infectious and Parasitic Diseases, University of São Paulo School of Medicine, Av. Dr. Enéas Carvalho de Aguiar, 470, São Paulo 05403 000, Brazil.
ACS Omega. 2025 May 1;10(18):18444-18456. doi: 10.1021/acsomega.4c10784. eCollection 2025 May 13.
Neglected tropical diseases as Chagas disease (CD) affect more than eight million people, mainly in the Americas, causing fatal cardiovascular outcomes. Relying on two old, toxic, and low efficacy drugs for treatment, there is an urgent need for new candidates. Comprising a high chemodiversity, marine bacteria are a rich source of small molecules with potential against human pathogens. Cultivation-based strategies of bacteria, such as the one strain many compounds (OSMAC) approach, have proven to be a simple and promising tool for drug discovery, with the ability to stimulate the expression of cryptic genes in microorganisms. In this study, using the OSMAC, we evaluated the potential of the marine bacteria to produce anti- compounds with higher potency. The was cultivated under different conditions, subdivided into four groups, as nutritional, physical, biological, and chemical alterations. For comparisons, the extract obtained from the bacteria in Marine Broth (static) at 25 °C was used as a control and resulted in an EC value of 28 μg/mL against the trypomastigotes. The physical alterations proved to be the most effective approach to improve the potency of metabolites, resulting in EC values between 3 and 26 μg/mL. The cultivation in Marine Agar potentiated the antitrypanosomal metabolites by 8.4-fold. When exposed to cobalt-60 γ radiation (0.5 kGy), the bacteria produced metabolites with 2-fold higher antitrypanosomal potency. The nutritional alterations resulted in potent metabolites, with EC values between 11 and 18 μg/mL, while biological alterations resulted in EC values between 11 and 28 μg/mL. Addition of and antigens and co-cultivation with , enhanced by 2-fold the potency. Chemical elicitors such as DMSO and EtOH demonstrated no improvements for cultivation. The chemical profile of was analyzed using NMR and UHPLC-ESI-HR-MS/MS and processed using the GNPS platform, which led to the annotation of nucleosides, dipeptides, steroids, and fatty acid derivatives. These findings confirmed that the OSMAC approach yielded not only distinct antitrypanosomal activities but also distinct metabolomic profiles in that could be exploited for drug discovery studies for Chagas disease.
恰加斯病（CD）等被忽视的热带病影响着超过800万人，主要在美洲，可导致致命的心血管疾病。由于治疗依赖于两种老旧、有毒且疗效不佳的药物，因此迫切需要新的候选药物。海洋细菌具有高度的化学多样性，是具有抗人类病原体潜力的小分子的丰富来源。基于培养的细菌策略，如“一个菌株多种化合物”（OSMAC）方法，已被证明是一种简单且有前景的药物发现工具，能够刺激微生物中隐秘基因的表达。在本研究中，我们使用OSMAC评估了海洋细菌产生高效抗化合物的潜力。该细菌在不同条件下培养，分为营养、物理、生物和化学改变四组。为作比较，以在25℃的海洋肉汤（静置）中培养的细菌提取物作为对照，其对锥鞭毛体的EC值为28μg/mL。事实证明，物理改变是提高该细菌代谢产物效力的最有效方法，EC值在3至26μg/mL之间。在海洋琼脂中培养使抗锥虫代谢产物增强了8.4倍。当暴露于钴 - 60γ辐射（0.5 kGy）时，该细菌产生的代谢产物抗锥虫效力提高了2倍。营养改变产生了高效的代谢产物，EC值在11至18μg/mL之间，而生物改变产生的EC值在11至28μg/mL之间。添加特定抗原并与其他生物共培养使效力提高了2倍。化学诱导剂如二甲基亚砜（DMSO）和乙醇（EtOH）对该细菌培养无改善作用。使用核磁共振（NMR）和超高效液相色谱 - 电喷雾电离 - 高分辨率质谱/质谱（UHPLC - ESI - HR - MS/MS）分析了该细菌的化学图谱，并使用全球天然产物社会分子网络（GNPS）平台进行处理，从而鉴定出核苷、二肽、类固醇和脂肪酸衍生物。这些发现证实，OSMAC方法不仅产生了不同的抗锥虫活性，而且在该细菌中产生了不同的代谢组学图谱，可用于恰加斯病的药物发现研究。
