Evaluating the multitargeted potency of Pixuvri against cell cycle regulation proteins in cervical cancer

Abstract

Cervical cancer, primarily caused by persistent infection with high-risk human papillomavirus strains, leads to abnormal cell growth in the cervix. Globally, it accounts for over 600,000 new cases and 340,000 deaths annually, with the highest burden in low- and middle-income countries due to limited screening and vaccination. Early detection is challenging as initial stages are asymptomatic, while advanced cases are challenging to treat. Current options, including surgery, radiotherapy, and chemotherapy, face issues like toxicity, limited efficacy, recurrence, and drug resistance caused by tumor heterogeneity and adaptive mechanisms. Multitargeted drug design offers a solution by modulating multiple cancer pathways, enhancing efficacy, minimizing resistance, and reducing side effects. In this study, we screened Selleckchem approved library against cervical cancer proteins that regulate the cell cycle, particularly during mitosis and cell division (PDBIDs: 2VFX, 2WVI, 3KND, 4N14) using HTVS, SP, and XP docking followed by MMGBSA post-processing. Pixuvri (Pixantrone Maleate) emerged as the top candidate with docking scores of -5.234 to -9.218 kcal/mol and MMGBSA scores of -39.22 to -53.87 kcal/mol. Pixuvri is approved for non-Hodgkin lymphoma and exhibits minimal cardiotoxicity compared to anthracyclines. Interaction fingerprints highlighted key residues (4GLN, 4GLU, 3TRP), while pharmacokinetics, DFT computations, and WaterMap hydration site analysis supported its potential. Molecular dynamics (100 ns, NVT ensemble at 300K) validated stability by deviation and fluctuation studies and found many interactions to stabilize the complex, with binding free energy computations confirming its affinity. While the results support Pixuvri's repurposing for cervical cancer, experimental validation is essential for clinical application.

Keywords: Pixuvri; WaterMap; cervical cancer; drug resistance; multitargeted drugs.

Figure 1

The workflow of the complete study to identify and validate the multitargeted drug candidate against cervical cancer proteins.

Figure 2

Prepared 3D structure of proteins and Ramachandran Plot for (A) 2VFX, (B) 4N14, (C) 2WVI, and (D) 3KND.

Figure 3

Ligand Interaction Diagram of docked poses in 2D for showing the interacting residues and bond types and 3D for fit in the pocket for (A) 2VFX, (B) 4N14, (C) 2WVI, and (D) 3KND.

Figure 4

Molecular Interaction Fingerprints of Pixuvri in complex with (A) 2VFX, (B) 4N14, (C) 2WVI, and (D) 3KND. The colored plot shows the N to C terminal of the protein. The right-side bar shows the count of ligand interactions, evidence of 2VFX having the highest interactions, and the upper-side bar shows the count of residue interactions.

Figure 5

The Density Functional Theory (DFT) results of Pixuvri are shown for plotted various energies, including the relative energy of the compound.

Figure 6

The 5ns WaterMap results in a ligand interaction diagram in 2D showing interacting residues and bond types along with the hydration sites of Pixuvri in complex with (A) 2VFX, (B) 4N14, (C) 2WVI, and (D) 3KND.

Figure 7

Showing the Root Mean Square Deviation for Pixuvri in complex with (A) 2VFX, (B) 4N14, (C) 2WVI, and (D) 3KND and Root Mean Square Fluctuation for the Pixuvri in complex with (E) 2VFX, (F) 4N14, (G) 2WVI, and (H) 3KND.

Figure 8

Simulation Interaction Diagram produced after 100ns MD Simulation for Pixuvri in complex with (A) 2VFX, (B) 4N14, (C) 2WVI, and (D) 3KND. The figure also depicts the interaction and types, and the interaction count is plotted separately to clarify the bond types.

Figure 9

The total complex energy and binding free energy of Pixuvri in complex with 2VFX, 4N14, 2WVI, and 3KND computed with the Molecular Mechanics Generalized Born Surface Area computations on produced trajectories of 100nd MD simulations.