Evaluation of Apremilast as a Potential Inhibitor of Activated Factor XII: In-Vitro and In-Silico Approaches

Madkhali, Hassan A.; Ansari, Mohd Nazam; Riadi, Yassine; Ganaie, Majid A.; Hamadi, Abdullah Y.; Alhawiti, Naif M.

doi:10.48309/ajca.2026.566309.2009

Evaluation of Apremilast as a Potential Inhibitor of Activated Factor XII: In-Vitro and In-Silico Approaches

Document Type : Original Research Article

Authors

Hassan A. Madkhali ¹

Mohd Nazam Ansari ²

Yassine Riadi ³

Majid A. Ganaie ⁴

Abdullah Y. Hamadi ⁵

Naif M. Alhawiti ⁶

¹ Department of Pharmacology and Toxicology, College of Pharmacy, Prince Sattam Bin Abdulaziz University, Al-Kharj, Saudi Arabia

² Department of Pharmacology & Toxicology, College of Pharmacy, Prince Sattam Bin Abdulaziz University, Al-Kharj 11942, Saudi Arabia

³ Department of Pharmaceutical Chemistry, College of Pharmacy, Prince Sattam Bin Abdulaziz University, 11942 Al-Kharj, Saudi Arabia

⁴ College of Dentistry and Pharmacy, Buraydah Colleges, Buraydah, Saudi Arabia

⁵ Faculty of Applied Medical Sciences, Medical Laboratory Technology, University of Tabuk, Tabuk, Saudi Arabia

⁶ King Abdullah International Medical Research Center, Ministry of National Guard for Health Affairs, Riyadh, Saudi Arabia

https://doi.org/10.48309/ajca.2026.566309.2009

Abstract

This study aimed to evaluate the inhibitory effects of apremilast on activated coagulation factor XII (FXIIa) using chromogenic enzyme assays and to examine molecular interactions between apremilast and benzamidine with the target protein 6I63 (FXIIa) through in silico techniques including docking, structural analyses, and dynamic simulations. The interaction strength of these complexes, focusing on hydrogen bonds, Root Mean Square Deviation (RMSD), Root Mean Square Fluctuation (RMSF), and Solvent Accessible Surface Area (SASA), was analyzed. Additionally, the selectivity of apremilast toward FXIIa over other related serine proteases (FIXa, FXa, FXIa, and plasma kallikrein) was determined using chromogenic assays. The findings indicated that apremilast has a significant inhibitory effect on FXIIa, while no notable effects were observed on the other proteases, suggesting selectivity for FXIIa. Furthermore, apremilast showed stronger binding affinity with 6I63 (−6.9 kcal/mol) compared to benzamidine (−5.3 kcal/mol), indicating a more stable and specific interaction. Apremilast induced greater compactness and stability of the complex, with lower RMSD and SASA values, whereas benzamidine favored a more flexible and less stable interaction. These results highlight distinct mechanisms by which each ligand interacts with FXIIa and provide insights into their therapeutic potential, suggesting apremilast could be effective in treating FXIIa-related diseases. In addition, ADME analysis revealed that benzamidine has better solubility, intestinal absorption, and excretion, while apremilast exhibits superior oral absorption and stronger inhibition of cytochrome P450 enzymes. Toxicity studies showed both compounds are mutagenic. Taken together, apremilast could serve as a lead compound for developing new therapeutic agents targeting FXIIa-related diseases.

Graphical Abstract

Evaluation of Apremilast as a Potential Inhibitor of Activated Factor XII: In-Vitro and In-Silico Approaches

Keywords

Factor XIIa

Apremilast

In vitro

In silico

Dynamic molecular

SASA

Subjects

Pharmaceutical chemistry

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