Ann Dermatol Vol. 25, No. 1, 2013 http://dx.doi.org/10.5021/ad.2013.25.1.5 ORIGINAL ARTICLE Preventive Effects of Multi-Lamellar Emulsion on Low Potency Topical Steroid Induced Local Adverse Effect Geun Dong Sul, Hyun Jung Park1, Jong Hwan Bae1, Keum Duck Hong1, Byeong Deog Park1, Jaesun Chun, Se Kyoo Jeong1, Seung Hun Lee2, Sung Ku Ahn3, Hyun Jung Kim4 Department of Biology Education, Korea National University of Education, Cheongwon, 1Research Division, NeoPharm Co., Ltd., Daejeon, 2Department of Dermatology, Yonsei University College of Medicine, Seoul, 3Department of Dermatology, Yonsei University, Wonju College of Medicine, Wonju, 4Department of Dermatology, Atopy and Asthma Center, Seoul Medical Center, Seoul, Korea Background: Topical steroid treatment induces diverse local adverse effects, including permeability barrier function Wand systemic adverse effects. Several approaches have impairment, were partially prevented by MLE formulation. been tried to reduce the steroid-induced adverse effects. Conclusion: Topical desonide formulation using MLE as a Simultaneous application of physiological lipid mixture is vehicle showed a better drug delivery with increased also suggested. Objective: Novel vehicles for topical epidermal retention. MLE also partially prevented the glucocorticoids formulation were evaluated for the efficacy steroid-induced side effects, such as skin barrier impairment. of reducing side-effects and the drug delivery properties of (Ann Dermatol 25(1) 5∼11, 2013) desonide, a low potency topical steroid. Methods: Trans- cutaneous permeation and skin residual amount of desonide -Keywords- were measured using Franz diffusion cells. The in vivo Epidermal permeability barrier function, Multilamellar anti-inflammatory activity was evaluated using murine emulsion, Skin permeation, Topical glucocorticoids model. Results: Topical steroids formulation containing de- sonide, in either cream or lotion form, were prepared using multi-lamellar emulsion (MLE), and conventional desonide INTRODUCTION formulations were employed for comparison. MLE for- mulations did not affect the anti-inflammatory activity of the Topical glucocorticoid therapy is one of the most com- desonide in phobol ester-induced skin inflammation model, monly used immunosuppressive or anti-inflammatory drugs. compared with conventional formulations. While the pene- Even with the invaluable therapeutic efficacies, however, trated amounts of desonide were similar for all the tested inevitable local side effects including skin atrophy, striae, formulations at 24 hours after application, the increased lag telangiectasis, prupura, acneform eruption, perioral der- time was observed for the MLE formulations. Interestingly, matitis, and steroid-induced rosacea impede the long-term residual amount of desonide in epidermis was significantly use of steroids for chronic skin diseases such as atopic higher in lotion type MLE formulation. Steroid-induced dermatitis. Recently, it was also reported that both the long-term and short-term application of topical steroids Received August 12, 2010, Revised October 26, 2011, Accepted for can compromise the skin barrier function, mainly by inhi- publication October 28, 2011 biting the epidermal lipid synthesis1,2. Corresponding author: Hyun Jung Kim, Department of Dermatology, Various strategies to minimize the steroid-induced side Atopy and Asthma Center, Seoul Medical Center, 156 Sinnae-ro, Jungnang-gu, Seoul 131-865, Korea. Tel: 82-2-2276-7891, Fax: 82-2- effects have been suggested, including modification of 3-5 6 2276-7438, E-mail: [email protected] application schedules , designing new steroid molecules , This is an Open Access article distributed under the terms of the and adjunctive use of other drugs with anti-atrophogenic Creative Commons Attribution Non-Commercial License (http:// activity7. Use of new vehicles for topical steroids is also creativecommons.org/licenses/by-nc/3.0) which permits unrestricted suggested as an effective way of reducing the side-effects non-commercial use, distribution, and reproduction in any medium, of steroids, as well as improving the transcutaneous de- provided the original work is properly cited. Vol. 25, No. 1, 2013 5 GD Sul, et al livery and epidermal release profile. It has been reported same 0.05% desonide were used for comparison. that a hydrogel formulation containing desonide, a low Preparation of test formulations potency corticosteroid, had an improved risk-benefit ratio in atopic dermatitis patients8,9. Recently, we have shown In order to prepare MLE vehicle, the lipid components that a multi-lamellar emulsion (MLE), as a vehicle for clo- (pseudoceramide, stearic acid, and cholesterol), oils, co- betasol 17-propionate, is effective for preventing the ste- mmon base (cetanol, 2-octyldodecanol, squalane, and roid-induced skin atrophy and epidermal permeability fatty acid triglyceride) and emulsifiers (glyceryl mono- barrier function impairment10. stearate, POE(15) glyceryl monostearate) were mixed and In the present study, the effects of MLE as a vehicle for melted at 85 to 90oC. The desonide was dissolved in 1, topical steroids is further investigated. Desonide, a low 3-butylene glycol and then slowly added with vigorous potency glucocorticoid, is formulated into MLE, and the agitation by homomixer (T.K. Homomixer Mark II f-model, anti-inflammatory activity and transcutaneous delivery Tokyo Co., Ltd., Tokyo, Japan) at 70oC. After emulsifying, were measured. Steroid-induced side effects on skin atro- the mixture was cooled down to the room temperature. phy and skin barrier function were also evaluated in All formulations were prepared containing 0.05% deso- animal model. nide of the final content (Table 1). Detailed preparation scheme was previously reported12. MATERIALS AND METHODS In vivo efficacy measurement Materials Female hairless mouse (8 weeks old) and male ICR mice Stearic acid, cholesterol, and fatty acid triglyceride were (4 weeks old) were purchased from Orient Bio Co. purchased from Junsei Chemical Co. (Tokyo, Japan); and (Seongnam, Korea) and kept under controlled humidity (50∼ glyceryl monostearate and POE(15) glyceryl monostearate 60%) and temperature (23∼25oC) with a 12 hr/12 hr were from Nihon Emulsion Co (Tokyo, Japan). Cetanol (light/dark) cycle. All the animal experiments described (Kao Co., Tokyo, Japan) and squalane (Kishimoto Co., were approved by the Institutional Review Board of Hyogo, Japan) were used for the preparation of emulsions. Yonsei University College of Medicine. Synthesis of pseudoceramide (myristyl/palmityl oxostearamide/ In order to observe the steroid-induced skin atrophy, arachamide MEA: C34H67NO3/C36H71NO3/C38H75NO3=11 ： tested formulations were applied to the entire dorsal skin 42：47%) and its chemical characteristics were described of hairless mouse twice a day for 3 days. Skin double-fold in a previous report11. Desonide was obtained from Hubei thickness was measured using digital calipers (Mitutoyo Gedian Humanwell Pharmaceutical Co., Ltd. (Hubei, Co., Ltd., Kawasaki, Japan). Epidermal permeability barrier China). Commercial cream and lotion products containing function and skin hydration were evaluated by measuring Table 1. The main components of tested formulations Conventional lotion Conventional cream MLE/desonide cream MLE/desonide lotion Desonide (0.05%) Sorbic acid Methyl hydroxybenzoate Pseudo-stratum corneum Pseudo-stratum corneum lipid (1.6%) lipid (2.3%) Potassium sorbate Propyl hydroxybenzoate Methyl hydroxybenzoate Methyl hydroxybenzoate Propylene glycol Propylene glycol Propyl hydroxybenzoate Propyl hydroxybenzoate Citric acid Citric acid Butylenes glycol Butylenes glycol Sodium hydroxide Sodium hydroxide Arginine Arginine Polysorbate 60 Sorbitan monostearate PEG-15 glyceryl stearate PEG-15 glyceryl stearate Emulsifying wax Self-emulsifying glyceryl Glyceryl monosterate Glyceryl monosterate monosterate Isopropyl palmitate Sodium lauryl sulphate Cetanol Cetanol Stearic acid Light liquid paraffin Squalane Squalane Synthetic beeswax Cetyl alcohol Liquid paraffin Liquid paraffin Propyl gallate Stearyl alcohol BHT BHT Purified water Tetrasodium EDTA Purified water Purified water Purified water MLE: multi-lamellar emulsion, EDTA: ethylenediaminetetraacetic acid, BHT: butylated hydroxytoluene. 6 Ann Dermatol The Preventive Effect of MLE Cream to Steroid Side Effect trans-epidermal water loss (TEWL) and skin capacitance, hydrated in 0.05% sodium chloride solution for 30 respectively. Measurement of TEWL and skin capacitance minutes and cut into appropriate pieces. The skin sections was performed using Tewameter TM 300 (Courage & were mounted on surface areas of 0.785 cm2 occluded Khazaka, Cologne, Germany) and Corneometer CM 820 Franz diffusion cells and the acceptor compartment was (Courage & Khazaka), respectively. filled with phosphate buffer pH 7.4. The epidermal side of Anti-inflammatory activity of desonide formulation was the skin was exposed to ambient environment and evaluated using mice ear model, and the skin inflamma- allowed to equilibrate for 1 hour before experiment. After tion was induced by repeated treatment of 12-O-tetrade- equilibration, 40 mg of each formulation was placed on canoylphorbol-13-acetate (TPA) for 7 days. TPA (1 μg/ skin surface using a spatula and receptor solution was 20 μl acetone) was applied to the mouse ear (day 0). The maintained at 37±0.5oC and stirred continuously at 600 tested formulations were then topically applied to the rpm. The receptor solution (4 ml) was taken