Abstract：
BACKGROUND: Herbal drugs when used in combination with chemotherapeutic drugs can reduce the side effects and increase the efficacy by acting on multiple targets. Andrographolide (AG), a diterpene lactone isolated from Andrographis paniculata Nees, is a bioactive compound with anticancer potential, and 5-fluorouracil (FU), a pyrimidine analogue, is used in the treatment of cancer. Both drugs are used to formulate combination nanoformulation to increase absorption, thereby increasing their oral bioavailability.
OBJECTIVE: The study aimed to develop and validate stability indicating simultaneous HPTLC method for quantification of FU and AG in combination nanoformulation along with in silico docking and network pharmacology analysis to understand the interaction between the drugs and cancer targets.
METHODS: Chromatographic separation was performed using mobile phase chloroform: methanol: formic acid (9: 0.5: 0.5, v/v/v) on HPTLC silica plates 60 F254 as a stationary phase using UV-Vis detector and HPTLC scanner at 254 nm. Further, in silico docking analysis was performed to predict the binding affinity of AG and FU with different proteins and network pharmacology to find out the exact biomolecular relationship of AG and FU in alleviating cancer.
RESULTS: The data from the calibration curve showed a good linear regression relationship with r² = 0.9981 (FU) and r² = 0.9977 (AG) in the concentration range of 0.1-2.0 μg/mL. The developed method was validated according to the ICH guidelines. Stability studies showed changes in peak patterns and areas. Bioinformatic and network pharmacology analyses of AG and FU with target proteins and genes associated with cancer play a multimechanistic role in alleviating cancer.
CONCLUSIONS: The developed method has been concluded to be robust, simple, precise, reproducible, accurate, and stability indicating for simultaneous quantification of AG and FU, and the molecular interaction studies have further indicated that the combination nanoformulation of AG and FU could be effective against cancer.
OBJECTIVE: The study aimed to develop and validate stability indicating simultaneous HPTLC method for quantification of FU and AG in combination nanoformulation along with in silico docking and network pharmacology analysis to understand the interaction between the drugs and cancer targets.
METHODS: Chromatographic separation was performed using mobile phase chloroform: methanol: formic acid (9: 0.5: 0.5, v/v/v) on HPTLC silica plates 60 F254 as a stationary phase using UV-Vis detector and HPTLC scanner at 254 nm. Further, in silico docking analysis was performed to predict the binding affinity of AG and FU with different proteins and network pharmacology to find out the exact biomolecular relationship of AG and FU in alleviating cancer.
RESULTS: The data from the calibration curve showed a good linear regression relationship with r² = 0.9981 (FU) and r² = 0.9977 (AG) in the concentration range of 0.1-2.0 μg/mL. The developed method was validated according to the ICH guidelines. Stability studies showed changes in peak patterns and areas. Bioinformatic and network pharmacology analyses of AG and FU with target proteins and genes associated with cancer play a multimechanistic role in alleviating cancer.
CONCLUSIONS: The developed method has been concluded to be robust, simple, precise, reproducible, accurate, and stability indicating for simultaneous quantification of AG and FU, and the molecular interaction studies have further indicated that the combination nanoformulation of AG and FU could be effective against cancer.
摘要：
背景：草药与化疗药物联合使用时，可以通过作用于多个靶标来减少副作用并提高疗效。穿心莲内酯(AG),从穿心莲中分离出的二萜内酯，是一种具有抗癌潜力的生物活性化合物，和5-氟尿嘧啶（FU），嘧啶类似物,用于治疗癌症。两种药物都用于配制组合纳米制剂以增加吸收，从而增加其口服生物利用度。
目的：该研究旨在开发和验证稳定性，表明同时使用HPTLC方法定量纳米制剂中的FU和AG，以及计算机对接和网络药理学分析，以了解药物与癌症靶标之间的相互作用。
方法:使用流动相氯仿:甲醇:甲酸(9:0.5:0.5,v/v/v)在HPTLC二氧化硅板60F254上作为固定相,使用UV-Vis检测器和HPTLC扫描仪在254nm下进行色谱分离。Further,进行了计算机对接分析，以预测AG和FU与不同蛋白质和网络药理学的结合亲和力，以找出AG和FU在减轻癌症中的确切生物分子关系。
结果：来自校准曲线的数据显示出良好的线性回归关系，在0.1-2.0μg/mL的浓度范围内，r²=0.9981（FU）和r²=0.9977（AG）。所开发的方法根据ICH指南进行了验证。稳定性研究显示峰模式和面积的变化。AG和FU的生物信息学和网络药理学分析与癌症相关的靶蛋白和基因在减轻癌症中起着多机制作用。
结论：所开发的方法被认为是稳健的，简单,精确，可重复,准确,以及同时定量AG和FU的稳定性，分子相互作用研究进一步表明，AG和FU的组合纳米制剂可以有效对抗癌症。
目的：该研究旨在开发和验证稳定性，表明同时使用HPTLC方法定量纳米制剂中的FU和AG，以及计算机对接和网络药理学分析，以了解药物与癌症靶标之间的相互作用。
方法:使用流动相氯仿:甲醇:甲酸(9:0.5:0.5,v/v/v)在HPTLC二氧化硅板60F254上作为固定相,使用UV-Vis检测器和HPTLC扫描仪在254nm下进行色谱分离。Further,进行了计算机对接分析，以预测AG和FU与不同蛋白质和网络药理学的结合亲和力，以找出AG和FU在减轻癌症中的确切生物分子关系。
结果：来自校准曲线的数据显示出良好的线性回归关系，在0.1-2.0μg/mL的浓度范围内，r²=0.9981（FU）和r²=0.9977（AG）。所开发的方法根据ICH指南进行了验证。稳定性研究显示峰模式和面积的变化。AG和FU的生物信息学和网络药理学分析与癌症相关的靶蛋白和基因在减轻癌症中起着多机制作用。
结论：所开发的方法被认为是稳健的，简单,精确，可重复,准确,以及同时定量AG和FU的稳定性，分子相互作用研究进一步表明，AG和FU的组合纳米制剂可以有效对抗癌症。
