Improved Subcutaneous Delivery of Ketoconazole Using EpiDerm and HSPiP Software-Based Simulations as Assessed by Cell Viability, Cellular Uptake, Permeation, and Hemolysis In Vitro Studies

Afzal Hussain; Obaid Afzal; Mohammad A Altamimi; Abdulmalik Saleh Alfawaz Altamimi; Mohhammad Ramzan; Mohd Zaheen Hassan; Wael A Mahdi; Thomas J Webster

doi:10.1021/acsomega.2c06001

Improved Subcutaneous Delivery of Ketoconazole Using EpiDerm and HSPiP Software-Based Simulations as Assessed by Cell Viability, Cellular Uptake, Permeation, and Hemolysis In Vitro Studies

ACS Omega. 2022 Nov 9;7(46):42593-42606. doi: 10.1021/acsomega.2c06001. eCollection 2022 Nov 22.

Authors

Afzal Hussain¹, Obaid Afzal², Mohammad A Altamimi¹, Abdulmalik Saleh Alfawaz Altamimi², Mohhammad Ramzan^{3

4}, Mohd Zaheen Hassan⁵, Wael A Mahdi¹, Thomas J Webster^{6

7

8}

Affiliations

¹ Department of Pharmaceutics, College of Pharmacy, King Saud University, Riyadh11451, Saudi Arabia.
² Department of Pharmaceutical Chemistry, College of Pharmacy, Prince Sattam Bin Abdulaziz University, Al-Kharj11942, Saudi Arabia.
³ Department of Pharmaceutics, School of Pharmaceutical Sciences, Lovely Professional University, Jalandhar-Delhi GT Road, Phagwara144411, Punjab, India.
⁴ Department of Pharmaceutics, University Institute of Pharmaceutical Sciences, Panjab University, Chandigarh160014, Punjab, India.
⁵ Department of Pharmaceutical Chemistry, College of Pharmacy, King Khalid University, Abha61441, Saudi Arabia.
⁶ School of Health and Biomedical Engineering, Hebei University of Technology, Tianjin065000, China.
⁷ School of Engineering, Saveetha University, Chennai602117, India.
⁸ Materials Science Program, UFPI, Teresina64064, Brazil.

Abstract

Ketoconazole (KETO) is the drug of choice to control local, systemic, and resistant types of fungal infections. Subcutaneous (sub-Q) delivery offers several benefits. The present study investigated the sub-Q delivery of KETO using HSPiP software based on optimized concentrations of dimethylacetamide (DMA) in binary solvents (DMA + water), in vitro cellular uptake (J774A.1) assays, cellular toxicity (L929), and in vitro hemolysis studies. Results showed that the estimated permeation coefficient (9.6 × 10^-3 cm/h) and diffusion coefficient (3.9 × 10^-3 cm²/h) of KETO (22.3 mg) in KF3 (300 mg of DMA + water) across EpiDerm were relatively higher as compared to the other formulations [KF1 (11.2 and 150 mg as KETO and DMA, respectively) and KF2 [(22.3 and 300 mg as KETO and DMA, respectively)] due to the increased content of DMA and KETO. HSPiP simulated and predicted the impact of constant and variable diffusion coefficients on the percent drug absorption across EpiDerm and the time needed to achieve equilibrium. The concentration-dependent diffusion coefficient fed into HSPiP predicted that the drug absorption and permeation values were linearly dependent on the square root of time. The HSPiP predicted permeation flux values from KF3, KF2, and KF1 across the EpiDerm were 4.07 × 10^-6, 4.01 × 10^-6, and 1.1 × 10^-6 g/cm²/s, respectively, at respective D range values. The selected K30G (324 mOsm/Kg) showed an optimal pH (6.9) and minimum drug loss (0.01%) over a period of 1 month at room temperature. KG30 was found to be less toxic to normal L292 cells and caused maximum cytotoxicity to candida cells residing within infected macrophage cells (J774A.1 incubated for 24 h), which was attributed to the slow diffusion of K30G compared to DS (the drug solution with an equivalent concentration). KG30 elicited substantial internalization with candida albicans (MTCC 4748) compared to the control group (24 h). Lastly, in vitro hemolysis studies (1 and 5 μg/mL) corroborated the safety of K30G for sub-Q delivery. Therefore, this new formulation and approach for delivering KETO is a promising alternative to conventional products to control fungal infections and, thus, should be further studied in vivo.