Repurposing levocetirizine hydrochloride loaded into cationic ceramide/phospholipid composite (CCPCs) for management of alopecia: central composite design optimization, in- silico and in-vivo studies

Abstract

Levocetirizine hydrochloride (LVC) is an antihistaminic drug that is repurposed for the treatment of alopecia. This investigation is targeted for formulating LVC into cationic ceramide/phospholipid composite (CCPCs) for the management of alopecia. CCPCs were fabricated by ethanol-injection approach, through a central composite experiment. CCPCs were evaluated by inspecting their entrapment efficiency (EE%), polydispersity index (PDI), particle size (PS), and zeta potential (ZP). The optimum CCPCs were additionally studied by in-vitro, ex-vivo, in-silico, and in-vivo studies. The fabricated CCPCs had acceptable EE%, PS, PDI, and ZP values. The statistical optimization elected optimum CCPCs composed of 5 mg hyaluronic acid, 10 mg ceramide III, and 5 mg dimethyldidodecylammonium bromide employing phytantriol as a permeation enhancer. The optimum CCPCs had EE%, PS, PDI, and ZP of 88.36 ± 0.34%, 479.00 ± 50.34 nm, 0.377 ± 0.0035, and 20.20 ± 1.13 mV, respectively. The optimum CCPC maintained its stability for up to 90 days. It also viewed vesicles of tube shape via transmission electron microscope. The in-silico assessment resulted in better interaction and stability between LVC and vesicle components in water. The ex-vivo and in-vivo assessments showed satisfactory skin retention of LVC from optimum CCPCs. The histopathological assessment verified the safety of optimum CCPCs to be topically applied. Overall, the optimum CCPCs could be utilized as a potential system for the topical management of alopecia, with a prolonged period of activity, coupled with reduced LVC shortcomings.

Keywords: Alopecia; ceramide; drug discovery; in-silico study; industrial development; levocetirizine hydrochloride.

Figure 1.

Effect of formulation variables on EE% of LVC-CCPCs (A-B) and PS (C-D). Abbreviation: EE%: Entrapment efficiency percentage, PS: Particle size, LVC: levocetirizine hydrochloride and CCPCs; cationic ceramide/phospholipid composite.

Figure 2.

Effect of formulation variables on ZP of LVC-CCPCs (E-F) and desirability figure(G). Abbreviation: ZP: zeta potential; LVC: levocetirizine hydrochloride and CCPCs; cationic ceramide/phospholipid composite.

Figure 3.

Transmission electron micrographs of LVC-loaded CCPCs. Abbreviation: LVC: levocetirizine hydrochloride and CCPCs; cationic ceramide/phospholipid composite.

Figure 4.

Predicted docking pose of LVC-PC binding complex. Stick 3 D-representation of LVC (yellow) loaded on PC interface (green), in presence of several CCPCs additives; ceramide (cyan), DDAB (blue), Phytanriol (orange), and HA (magenta). All introduced formulation additives served as either direct or indirect connecting/supportive platforms for favored anchoring of LVC within the drugphospholipid complex. Polar interactions (hydrogen bond) are represented as black dashed lines. Abbreviation: HA: hyaluronic acid; DDAB: dimethyldidodecylammonium bromide; PC: phospholipid; LVC: levocetirizine hydrochloride and CCPCs; cationic ceramide/phospholipid composite.

Figure 5.

Time evolution conformational alterations of LVC-PC-CCPCs additive heterocomplex throughout an explicit MD simulation run at a 100% aqueous solvation system. The thermodynamic movements and stability of formulation components; LVC (yellow) loaded on PC interface (green), in presence of several CCPCs; Ceramide (cyan), DDAB (blue), Phytantriol (orange), and HA (magenta), were monitored over MD simulation trajectories being captured at different snapshots ① 0.2 ns, ② 0.4 ns; ③ 0.6 ns; ④ 0.8 ns; and ⑤ 1 ns. Polar interactions (hydrogen bond) are represented as black dashed lines. Abbreviation: HA: hyaluronic acid; DDAB: dimethyldidodecylammonium bromide; PC: phospholipid; LVC: levocetirizine hydrochloride and CCPCs; cationic ceramide/phospholipid composite; MD: molecular dynamic simulation.

Figure 6.

Overlay of LVC-PC-CCPCs additive heterocomplex across MD simulation frames (left panel) and molecular surface 3 D-representation of the inverted cone micellar configuration at 100% aqueous solvation system (right panel). Molecular surface and sticks 3 D representations were illustrated in colors being previously assigned for the optimized formulation components; yellow, green, cyan, blue, orange, and magenta for LVC, PC, Ceramide, DDAB, Phytantriol, and HA, respectively. Abbreviation: HA: hyaluronic acid; DDAB: dimethyldidodecylammonium bromide; PC: phospholipid; LVC: levocetirizine hydrochloride and CCPCs; cationic ceramide/phospholipid composite; MD: molecular dynamic simulation.

Figure 7.

Ex-vivo permeation profile of LVC from CCPCs, compared to its aqueous solution. Abbreviation: LVC: levocetirizine hydrochloride and CCPCs; cationic ceramide/phospholipid composite.

Figure 8.

In-vivo skin deposition profile of LVC from CCPCs, compared to its aqueous solution. Abbreviation: LVC: levocetirizine hydrochloride, and CCPCs; cationic ceramide/phospholipid composite.

Figure 9.

Light microscope photomicrographs showing histopathological sections (hematoxylin and eosin-stained) of rat skin in normal control (group I), rat skin treated with LVC solution (group II), and rat skin treated with optimum CCPCs (group III) with a magnification power of 16X to illustrate all skin layers (Left side) and magnification power of 40X to identify the epidermis and dermis (Right side). Abbreviation: LVC: levocetirizine hydrochloride, and CCPCs; cationic ceramide/phospholipid composite.