Department of Pharmaceutical Chemistry, College of Pharmacy, Najran University, Najran, Kingdom of Saudi Arabia.
Department of Pharmacy, Faculty of Biological Sciences, University of Malakand, Chakdara, 18000 Dir (L), KP, Pakistan.
BMC Complement Med Ther. 2021 Sep 24;21(1):239. doi: 10.1186/s12906-021-03411-1.
According to the recent global cancer statistics, breast cancer is the leading cause of deaths among women with 2.3 million new cases globally. Likewise, cervical cancer is also among the leading causes of mortality among women. Polygonum hydropiper is traditionally known for its cytotoxic effects and several bioactive cytotoxic compounds were isolated from it. This study was aimed to isolate potential anticancer compounds from its most potent fractions and evaluate their anticancer potentials.
Based on our earlier studies, active fractions including chloroform and ethyl acetate were subjected to column chromatography for isolation of compounds. Chemical structures of isolated compounds were confirmed via H NMR, C NMR, mass spectrometry. Purified compounds were tested for cytotoxicity against breast cancer cells (MCF-7), cervical cancer cells (HeLA) and NIH/3T3 fibroblasts cells cultures using MTT assy. Anti-angiogenic potentials of isolated compounds were evaluated via chorioallantoic membrane assay. Anti-tumor studies were done using Agrobacterium tumefaciens induced potato tumor assay. Furthermore, to understand the binding modes of Isolated compounds, molecular docking was performed against EGFR, HER2 and VEGFR using MOE as docking software.
Two bioactive compounds PH-1 (4-methyl-5-oxo-tetrahydrofuran-3-yl acetate) and PH-2 (methyl 4-hydroxy-3-methoxybenzoate) were purified from the active fractions. In cytotoxicity studies, PH-1 exhibited highest cytotoxicity against HeLA cells with 87.50% lethality at 1 mgmL concentration and LD of 60 µgmL. Likewise, PH-2 showed 82.33% cytotoxicity against HeLA cells with LD of 160 µgmL. Similarly, PH-1 and PH-2 exhibited LD of 170 and 380 µgmL respectively. Moreover, PH-1 and PH-2 were also very potent cytotoxic compounds against NIH/3T3 cells with 81.45 and 85.55% cytotoxicity at 1 mgL concentration and LD of 140 and 58 µgL respectively. Isolated compounds exhibited considerable anti-angiogenic potentials with IC of 340 and 500 µgL respectively for PH-1 and PH-2. In anti-tumor assay, PH-1 and PH-2 exhibited 81.15 and 76.09% inhibitions with LD of 340 and 550 µgL respectively. Both compounds selectively binds with EGFR and HER2 receptors with low binding energies. Both compounds exhibited stronger interactions with VEGFR through binding pocket residues Lys868, Val916 and Asp1046.
Both compounds cause considerable cytotoxicity against cancer cells. The anti-angiogenic and anti-tumor results suggests additional tumor suppressive properties. Docking analysis suggests that these compound not only has the ability to bind to EGFR and HER2 but also equally binds to VEGFR and may act as potential anti-angiogenic agents.
根据最近的全球癌症统计数据，乳腺癌是导致女性死亡的主要原因，全球有 230 万新发病例。同样，宫颈癌也是女性死亡的主要原因之一。水蓼传统上以其细胞毒性作用而闻名，已从其中分离出多种具有生物活性的细胞毒性化合物。本研究旨在从其最有效的部分中分离潜在的抗癌化合物，并评估其抗癌潜力。
根据我们之前的研究，包括氯仿和乙酸乙酯在内的活性部分通过柱色谱法进行分离化合物。通过 1H NMR、13C NMR、质谱法确定分离化合物的化学结构。用 MTT 法检测纯化化合物对乳腺癌细胞（MCF-7）、宫颈癌细胞（HeLA）和 NIH/3T3 成纤维细胞的细胞毒性。通过鸡胚尿囊膜试验评估分离化合物的抗血管生成潜力。使用根癌农杆菌诱导的马铃薯肿瘤试验进行抗肿瘤研究。此外，为了了解分离化合物的结合模式，使用 MOE 作为对接软件对 EGFR、HER2 和 VEGFR 进行了分子对接。
从活性部分中分离出两种生物活性化合物 PH-1（4-甲基-5-氧代四氢呋喃-3-基乙酸酯）和 PH-2（4-羟基-3-甲氧基苯甲酸甲酯）。在细胞毒性研究中，PH-1 对 HeLA 细胞表现出最高的细胞毒性，在 1mgmL 浓度下 87.50%的细胞致死率，LD 为 60µgmL。同样，PH-2 对 HeLA 细胞表现出 82.33%的细胞毒性，LD 为 160µgmL。同样，PH-1 和 PH-2 的 LD 分别为 170 和 380µgmL。此外，PH-1 和 PH-2 对 NIH/3T3 细胞也具有很强的细胞毒性，在 1mgL 浓度下的细胞毒性分别为 81.45%和 85.55%，LD 分别为 140µgmL 和 58µgL。分离出的化合物具有相当的抗血管生成潜力，PH-1 和 PH-2 的 IC 分别为 340 和 500µgL。在抗肿瘤试验中，PH-1 和 PH-2 表现出 81.15%和 76.09%的抑制作用，LD 分别为 340µgmL 和 550µgmL。两种化合物均选择性地与 EGFR 和 HER2 受体结合，结合能较低。两种化合物均通过结合口袋残基 Lys868、Val916 和 Asp1046 与 VEGFR 表现出更强的相互作用。
两种化合物均对癌细胞产生明显的细胞毒性。抗血管生成和抗肿瘤结果表明其具有额外的肿瘤抑制特性。对接分析表明，这些化合物不仅具有与 EGFR 和 HER2 结合的能力，而且还与 VEGFR 同等结合，并可能作为潜在的抗血管生成剂。
