Antioxidant and Cytotoxic Activity of the Aporphine Alkaloid (Boldine) Against DMH-induced Colorectal Carcinogenesis in Wistar Rats: An In silico, In vitro, and In vivo study.

Objective: The objective of this study is to investigate the role of the aporphine alkaloid boldine as a potential inhibitor of specific protein targets involved in colorectal cancer, using in silico docking and molecular dynamics simulation studies, and to evaluate its therapeutic potential in modulating the pathological progression of colorectal cancer. In this study, we evaluated the antioxidant and cytotoxic effects of boldine using in vitro and in vivo methods.

Methods: The 2-dimensional structure of boldine was retrieved from the PubChem database. Its interactions with colorectal cancer target proteins were analyzed using structures obtained from the RCSB Protein Data Bank (https://www.rcsb.org/), which provides the crystal structures of tubulin (PDB ID: 1Z2B), human NF- κB (1A3Q), human interleukin-2 (1M47), and EGFR-kinase. Molecular docking was then performed using Schrödinger software. Molecular docking and molecular dynamics (MD) simulations were conducted to evaluate Boldine's binding affinity and stability with colorectal cancer protein targets. On the Growmac platform, we performed the molecular simulation, and the simulation was only done with the highest docking score. Additionally, molecular dynamics was performed for 100 ns. Boldine was added to the colorectal cancer cell line (HCT116) at different doses, and the cytotoxic effects of the treatment were evaluated using cell viability assays. Furthermore, Boldine's capacity to scavenge reactive oxygen species (ROS) and modify the oxidative stress assay in vitro was used to assess its antioxidant capability. Boldine's in vivo anticancer effectiveness was examined using DMH-induced colorectal cancer in Wistar Rats. After Boldine was administered (100mg/Kg), Tumour progression, histological alteration, and oxidative stress markers were evaluated. The study aimed to ascertain how boldine affects tumor growth in DMH-inducedDMH colorectal carcinogenesis.

Results: Molecular docking revealed favorable binding interactions between boldine and key targets implicated in colorectal cancer. The in silico data supported the hypothesis that boldine modulates essential pathways associated with cancer development. Boldine exhibited a dose-dependent reduction in colorectal cancer cell viability, indicating potential cytotoxic effects. Furthermore, boldine demonstrated antioxidant properties by effectively scavenging reactive oxygen species (ROS) and modulating oxidative stress markers in vitro. In animal models, boldine administration resulted in a significant reduction in tumor growth. Histopathological examination revealed favorable changes in tumor morphology. Additionally, boldine demonstrated antioxidant effects in vivo by modulating oxidative stress markers.

Conclusion: According to this extensive study performed in silico, in vitro, and in vivo, boldine may be able to prevent CRC through its antioxidant and cytotoxic properties. These results encourage more research into boldine as a viable option for treating CRC. It is successfully done, and we can say that boldine is a valuable addition to the CRC treatment choice.