Instituto de Biotecnología, Facultad de Ingeniería, Universidad Nacional de San Juan, Av. Libertador General San Martin, 1109 O San Juan, Argentina; Consejo Nacional de Investigaciones Científicas y Técnicas (CONICET), CCT CONICET San Juan, Argentina.
Barcelona Institute for Global Health (ISGlobal), Hospital Clinic-University of Barcelona, 08036 Barcelona, Spain; Departament de Biologia, Sanitat i Medi Ambient, Facultat de Farmàcia i Ciències de l´Alimentació, Universitat de Barcelona, 08028 Barcelona, Spain.
Phytomedicine. 2023 Jun;114:154788. doi: 10.1016/j.phymed.2023.154788. Epub 2023 Mar 29.
Chagas disease (CD), caused by Trypanosoma cruzi, represents a health threat to around 20 million people worldwide. Side effects of benznidazole (Bzn) cause 15-20% of patients to discontinue their treatment. Evidence has increased in favor of the use of drug combinations to improve the efficacy and tolerance of the treatment. Natural products are well known to provide structures that could serve as new drugs or scaffolds for CD treatment. Spp of the Amaryllidoideae sub family of Amaryllidaceae family are known by their bioactives alkaloids, which have been reported by their antiparasitic activities.
To evaluate the anti-T. cruzi activity of the isolated alkaloid candimine (Cnd) from Hippeastrum escoipense Slanis & Huaylla; and to assess the combination effect between Cnd and Bzn against different life stages of T. cruzi parasites.
The chemical profile of H. escoipense alkaloids extract (AE-H. escoipense), including quantitation of Cnd was performed through GC/MS and UPLC-MS/MS techniques. Subsequently, Cnd was isolated using Shephadex LH-20. Then, the AE-H. escoipense and Cnd were tested against T. cruzi, (epimastigotes, trypomastigotes, and amastigotes) by in vitro proliferation and viability assays. The cytotoxicity was evaluated against Vero and HepG2 mammalian cells. The ultrastructural analysis was perform by transmission electron microscopy (TEM) and mitochondrial activity was carried out by MTT assay. Drug combination assay between Cnd and Bzn was evaluated using the Chou-Talalay method.
The AE-H. escoipense and Cnd showed high and specific anti-T. cruzi activity, comparable to Bzn. Cnd induces ultrastructural changes in T. cruzi, such as vacuolization, membrane blebs, and increased mitochondrial activity. Regarding the interaction between Cnd and Bzn, it generates synergism in the combinations of 0.25×IC in epimastigotes, 2×IC in trypomastigotes+amastigotes, and 0.25, 2, and 4×IC in amastigotes.
The synergism between Cnd and Bzn indicates that the combination at the concentration of 4×IC could be useful as an effective new therapy against CD in the chronic stage. Thus, Cnd isolated from the leaves of H. escoipense emerges as potential candidate for the development of a new drug for the treatment of CD.
恰加斯病（CD）由克氏锥虫引起，对全球约 2000 万人的健康构成威胁。苯并咪唑（Bzn）的副作用导致 15-20%的患者停止治疗。越来越多的证据表明，使用药物联合治疗可以提高治疗的疗效和耐受性。天然产物以提供可作为 CD 治疗新药物或支架的结构而闻名。石蒜科 Amaryllidaceae 亚科的 spp 以其生物活性生物碱而闻名，这些生物碱已被报道具有抗寄生虫活性。
评估 Hippeastrum escoipense Slanis & Huaylla 中分离的生物碱 candimine（Cnd）对 Trypanosoma cruzi 的抗活性；并评估 Cnd 与 Bzn 对 T. cruzi 不同生活阶段的组合效果。
通过 GC/MS 和 UPLC-MS/MS 技术对 H. escoipense 生物碱提取物（AE-H. escoipense）的化学特征进行了包括 Cnd 定量分析。随后，使用 Shephadex LH-20 分离 Cnd。然后，通过体外增殖和活力测定法测试 AE-H. escoipense 和 Cnd 对 T. cruzi（epimastigotes、trypomastigotes 和 amastigotes）的作用。用 MTT 测定法测定对 Vero 和 HepG2 哺乳动物细胞的细胞毒性。通过透射电子显微镜（TEM）进行超微结构分析，并进行线粒体活性测定。使用 Chou-Talalay 方法评估 Cnd 和 Bzn 之间的药物组合试验。
AE-H. escoipense 和 Cnd 对 T. cruzi 具有高特异性的抗活性，与 Bzn 相当。Cnd 诱导 T. cruzi 的超微结构变化，如空泡化、膜泡和增加的线粒体活性。关于 Cnd 和 Bzn 之间的相互作用，在 epimastigotes 中 0.25×IC、trypomastigotes+amastigotes 中 2×IC 以及 amastigotes 中 0.25、2 和 4×IC 的组合中产生协同作用。
Cnd 和 Bzn 之间的协同作用表明，在慢性期，浓度为 4×IC 的组合可能是治疗 CD 的有效新疗法。因此，从 H. escoipense 叶片中分离出的 Cnd 成为治疗 CD 的新药开发的潜在候选药物。
