Table1_Spectral, Anti-Inflammatory, Anti-Pyretic, Leishmanicidal, and Molecular Docking Studies, Against Selected Protein Targets, of a New Bisbenzylisoquinoline Alkaloid.doc

A new bisbenzylisoquinoline named as chondrofolinol (1) and four reported compounds (2–5) were isolated and characterized from the roots of Berberis glaucocarpa Stapf. Anti-inflammatory, anti-pyretic, and leishmanicidal studies were performed against carrageenan-induced paw edema, yeast-induced pyrexia, and the promastigotes of Leishmania tropica, respectively. The new compound significantly reduced the paw volume in carrageenan-induced paw edema and rectal temperature in yeast-induced pyrexia at 10 and 20 mg/ kg of body weight. Chondrofolinol caused almost 100% inhibition of the promastigotes of Leishmania tropica. All the compounds displayed minimal cytotoxicity against THP-1 monocytic cells. In order to ascertain the potential macromolecular targets of chondrofolinol responsible for the observed anti-inflammatory and anti-leishmanial activities, a molecular docking study was carried out on relevant protein targets of inflammation and Leishmania. Protein targets of human endoplasmic reticulum aminopeptidase 2 (ERAP2) and human matrix metalloproteinase-1 (MMP-1) for inflammation and protein targets of N-myristoyltransferase (NMT), tyrosyl-tRNA synthetase (TyrRS), and uridine diphosphate-glucose pyrophosphorylase (UGPase) for Leishmania major were selected after thorough literature search about protein targets responsible for inflammation and Leishmania major. Chondrofolinol showed excellent docking to ERAP2 and to MMP-1. The Leishmania major protein targets with the most favorable docking scores to chondrofolinol were NMT, TyrRS, and UGPase. The study indicated that bisbenzylisoquinoline and isoquinoline alkaloids possess anti-pyretic, anti-inflammatory, and anti-leishmanial properties with minimal cytotoxicity and therefore, need to be further explored for their therapeutic potential.

本研究从粉果小檗（Berberis glaucocarpa Stapf）的根部分离并表征了一种新型双苄基异喹啉（bisbenzylisoquinoline）类化合物，命名为软骨叶碱（chondrofolinol，1号），以及四个已知化合物（2~5号）。分别针对角叉菜胶诱导的足肿胀模型、酵母诱导的发热模型以及热带利什曼原虫前鞭毛体，开展了抗炎、解热及抗利什曼活性测试。在10与20 mg/kg体重的给药剂量下，该新化合物可显著降低角叉菜胶诱导足肿胀模型的足爪体积，以及酵母诱导发热模型的直肠体温。软骨叶碱对热带利什曼原虫前鞭毛体的抑制率近乎100%。所有受试化合物对THP-1单核细胞均仅表现出极低的细胞毒性。为明确软骨叶碱发挥抗炎及抗利什曼活性的潜在大分子靶点，本研究针对炎症相关与利什曼原虫相关的蛋白靶点开展了分子对接实验。通过全面检索与炎症及硕大利什曼原虫（Leishmania major）相关的蛋白靶点文献后，本研究选取了炎症相关的人内质网氨肽酶2（human endoplasmic reticulum aminopeptidase 2, ERAP2）、人基质金属蛋白酶-1（human matrix metalloproteinase-1, MMP-1），以及硕大利什曼原虫相关的N-肉豆蔻酰转移酶（N-myristoyltransferase, NMT）、酪氨酰-tRNA合成酶（tyrosyl-tRNA synthetase, TyrRS）与尿苷二磷酸葡萄糖焦磷酸化酶（uridine diphosphate-glucose pyrophosphorylase, UGPase）作为对接靶点。软骨叶碱与ERAP2、MMP-1均展现出极佳的结合对接效果。与软骨叶碱对接得分最优的硕大利什曼原虫蛋白靶点依次为NMT、TyrRS及UGPase。本研究表明，双苄基异喹啉类与异喹啉类生物碱具备解热、抗炎及抗利什曼活性，且细胞毒性极低，其治疗潜力有待进一步深入探索。