Lecithin Organogel as Matrix for Transdermal Transport of Drugs

H.WILLIMANN, P. WALDE*,P. L. Lusi*, A. G m N i G A * ,

AND

F. STROPPOLO*

Received May 21, 1991, from *nH-Zentnrm, lnstitut fiir Polymere, Ur!ive@Mstrasse6, CH-8092, Zurich, Switzerland, and *lnphsnam Ricerche Accepted for publication November 25, 1991. S.A. (Zambon Group), Cadempmo, Swrfzerlend.

Abstract 0 Organogels obtained by adding small amounts of water to a solution of lecithin in organic solvents were studied as matrices for the transdermal transport of drugs. Gels obtained from isopropyl palmitate and cyclooctane were used (molar ratios of water to lecithin of 3 and 12, respectively). Preliminarily histologicalstudies showed that the gels have no harmful effect when applied to the skin for prolonged periods. Data relative to the stability of the organogels with time are also presented. Scopolamine and broxaterol were used as model drugs, and the transdermal experiments were done with a Franz diffusion cell and human skin obtained from plastic surgery. The transport rate of scopolamine obtained with the lecithin gels was about one order of magnitude higher than that obtained with an aqueous solution of the drug at the same concentration. In contrast, the transport rates of scopolamine obtained with the microemulsion solution prior to gelation (molar ratio of water to lecithin, 0 ) were not different from those obtained with the gel. The same variations in transport rates were observed for broxaterol, in which case the flux through the skin was directly proportional to the concentration of drug in the gel. At a concentration of broxaterol of 75 mglmL in the donor gel, the flux was 47 pg * h- cm-*. Because preliminary results showed that transdermal transport is successful with amino acids and peptides also, it is concluded that lecithin gels may be efficient vehicles for the transdermal transport of various drugs.

needed to penetrate through the skin are not yet completely understood. Even if these prerequisites were known, however, chemically modifying all drugs of medicinal interest to make them transdermally active would not be practical. It is more reasonable to look for a carrier that interacts with the skin such that it allows various molecules to pass into the skin. In this paper, which follows a short preliminary report on the subject,12 we present the results of an investigation of lecithin gels as carriers for the transdermal transport of drugs. The general properties of lecithin gels (interaction with human skin, stability with time, and ability to solubilize drugs); the potential use of lecithin gels for the transdermal transport of scopolamine (known as a skin-penetrating substance); and the transdermal transport of the new drug broxaterol [(t)-3-bromo-a-[(tert-butylamino)methyll-5isoxazolemethanol]bronchodilator agent with selective p2 agonist activity,ls are discussed. Broxaterol hydrochloride is active in the treatment of asthma at oral doses of 0.25-0.5 mg,14J5 inhalation doses of 0.2-0.4 mg,16-19 and intravenous doses of 0.1-0.2 mg.20

Experimental Section
Lecithin organogels are readily obtained by adding a minimal amount of water to a solution of lecithin in organic solvents.1 Surprisingly high viscosities can be achieved (up to 10 000 poise) in various organic solvents1.2 addition of water to the small reverse lecithin micelles present initially brings about a monodimensional growth of long cylindrical giant micelles; above a certain critical concentration of lecithin, these spaghettilike micelles2 build a continuous entangled network, which, despite its dynamic character, has sufficient structural persistence to yield a high macroscopicviscosity.Biocompatible liquids, such as isopropyl palmitate (IPP) and other esters of this kind, can be used to form gels, which are then suitable for pharmaceutical and cosmetic formulations. Lecithin gels are isotropic and thermoreversible. At temperatures >40 C, they become liquids with much lower viscosity, and high-viscosity gels are again formed by cooling and stirring. These gels are also interesting because of their ability to host various guest molecules. Three types of molecules (lipophilic, hydrophilic, and amphoteric), including enzymes,6can be solubilized in the gels.1.7 The present work takes advantages of two basic properties of lecithin gels, biocompatibility and ability to solubilize drugs, and explores the possibility of using these drug-containing, biocompatible lecithin gels as a matrix for transdermal transport. Transdermal transport of drugs (i.e., the transport of pharmacologically active compounds through the skin into the blood vessels) is currently regarded as an important alternative to the classical ways of delivering drug, most notably peroral and percutaneous delivery by injection. The advantages of the transdermal route have been emphasized in the literature.gl1 The prerequisites of the chemical structure
OO22-3~9/92/0900-0871$02.50/0
0 1992. American Pharmaceutical Association

M a t e r i a l d o y b e a n lecithin (Epikuron 200) was from Lucas Meyer (Hamburg, Germany); isopropyl palmitate, cyclooctane, 4(2hydroxyethy1)piperazine-1-ethanesulfonic acid (HEPES), and sodium dihydrogen phosphate were from Fluka (Buchs, Switzerland); scopolamine hydrobromide and polyethylene glycol 4000 (PEG 4000) were from Merck (Darmstadt, Germany); pestradiol 17-acetate, pestradiol diacetate, pestradio1 17-valerate, pestradiol 17-enanthate, and pestradiol 17-cypionate were from Sigma (St. Louis, MO); acetonitrile (Chromasolv) was from Riedel-de Haen (Seelze, Germany); and broxaterol base, nifedipine, clonidine, and isosorbide dinitrate were from Inphanam Ricerche S.A.(Campedino, Switzerland).Water was deionized and doubly distilled in quartz glass. Human skin was received from the Plastic Surgery Division (Prof. M. Frei) of the University Hospital, Zurich, Switzerland. All skin samples were from chest sections of 30-40-year-old female patients. After surgery, the skin (epidermis and dermis) was carefully cleaned with water and stored at -40 C for a maximum of 150 days. Before each set of transdermal experiments, the skin sample was thawed at room temperature by soaking in 50 mM HEPES buffer (pH 6.8)for 15 min. The remaining fatty layers were carefully removed by forceps, and the skin sample was then cut into sections of -1 x 1 cm. Method+Preparatwn of Gels-Lecithin gels were prepared at room temperature by first dissolving lecithin in organic solvent (isopropyl palmitate or cyclooctane) and then adding, with magnetic stirring, the necessary amount of water to obtain the gel. The viscosity of the gel depends on the amount of water generally expressed as w,, the molar ratio of added water to lecithin (w, = [H,O]/[lecithin]). Dry Epikuron 200 lecithin usually contains -0.5-1 water molecule per lecithin molecule. Further details on the preparation of lecithin gels are given elsewhere.1.2 Commercial lecithins of lower purity do not form gels. High purity, asjudged by thin-layer chromatography, however, does not mean that we are dealing with only one compound chemically. In fact, natural lecithins are mixtures because of the heterogeneity of the fatty acid alkyl chains in positions 1 and 2 of the glycerol moiety.
Journal of Pharmaceutical Sciences I 871 Vol. 81, No. 9, September 1992

I
CH20H
Scopolamine

Br

OH
Broxaterol

The drug-containing gels were prepared by dissolving the drug in the lecithin dissolved in organic solvent and then adding water to induce gelation, as described earlier. The viecosity of the final gel formulation may be reduced, depending on the amount of solubilized drug. Transdermal In Vitro Studies-The passage of the solubilized drugs from the lecithin gel through human skin was studied with homemade, glass, vertical Franz diffusion cells.11-21In these cells, the skin samples were sandwiched between the upper donor compartment and the lower acceptor compartment. The circular area of skin that was in contact with the two compartments was 0.64 cm2. The acceptor compartment contained 5% PEG 4000 in 50 mM HEPES buffer (pH 6.8). PEG is often used in the acceptor solution,a mainly to increase the solubility of organic compounds. The cells were thermostated at 35 "C in an incubator (Reacti-Therm; Pierce, Wageningen, The Netherlands), and the acceptor solution was stirred with a magnetic stirrer at 400 rpm. Usually, 0.4 mL of gel containing the drug was placed in the donor compartment onto the skin, and the concentration of the drug in 5-20-pL aliquot8 of the acceptor solution (as reported in Figures 2-4) was determined by high-performance liquid chromatography (Series 4 system; Perkin-Elmer, Norwalk, CT; equipped with an LC 90 UV detector). For both scopolamine and broxaterol, a Hibar Lichrosorb CN column (5-pn mean particle size, 250-mm length, 4-mm id.) from E. Merck (Darmstadt, Germany) was used. Elution was performed with CH,CN:sodium phosphate buffer (70:30,20 mM, pH 4) at a flow rate of 1 mumin. The UV detector was set at 259 and 217 nm, for scopolamine and broxaterol, respectively. Quantifications were performed by comparing peak heights with calibration curves obtained with known amounts of drugs under identical analytical conditions. Histology-Thin sections of the skin were prepared and then stained with eosine B at the Dermatology Department of the University Hospital in Ziirich under the guidance of Dr. L. Bruckner.

Results and Discussion

General Properties of Lecithin Gels as Matrices for Transdermal Transport-The gel preparations used in this work were made with soybean lecithin (Epikuron 200) containing at least 95% phosphatidylcholine.22Isopropyl palmitate was chosen as a representative biocompatible solvent, and cyclooctane was chosen because it permits gels to be made at much larger values ofw,. Thew, values for gels with isopropylpalmitate and cyclooctane are 3 and 12, respedively.1 (The w, value of 12 is different from the value of 7 reported earlier for soybean lecithin from Sigma.) The dynamic shear viscosities at 25 "C and 0.5 rad/s for gels with isopropyl palmitate and cyclooctane are 170 and 3260 poise, respectively.1
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Lecithin gels kept at constant temperature are indefinitely stable in closed vials, without change of color or appearance. For example, the absorption spectrum between 250 and 500 nm of a 120mM soybean lecithin solution in cyclohexane (w, = 3)does not change within at least 1month. Even in open vials stored at room temperature, most of the lecithin gels remain stable for at least 30 days. For example, if one follows the Fourier transformed infrared spectrum of a 200 mM lecithin solution in isopropyl palmitate (w, = 2.5) during this time, no significant changes are observed between 3000 and 4000 cm-l, the OHstretching region of the spectrum (data not shown).This observation also indicates that the lecithin gel does not absorb a significant amount of humidity from the air during storage. The same is true for gels containing solubilized guest molecules, such as vitamin A palmitate or nifedipine. The solubility of drugs in gels tested to date has been satisfactory. Actually, the presence of lecithin in the organic solution often brings about an increase of solubility with respect to that observed in the neat solvent. Thus, broxaterol can be solubilized in isopropyl palmitate up to only 11mg/mL. In isopropyl palmitate with 200 mM lecithin, however, the solubility can be brought up readily to 75 mg/mL. Likewise, the solubility of the drug nifedipine in lecithin gels is considerably higher than the solubility of the drug in either water or isopropyl palmitate (data not shown). Likewise, we obtained good solubility of the following compounds in lecithin gels: pestradiol 17-acetate, pestradiol diacetate, @stradiol 17-valerate, pestradiol 17-enanthate, pestradiol 17cypionate, isosorbide dinitrate, and clonidine. The solubility of several amino acids and peptides is described elsewhere.6 Another important question preliminary to pharmaceutical applications is whether, and to what extent, lecithin gels are harmful to human skin. To partly clarify this point, we carried out a light microscopic investigation of human skin before and after treatment with either isopropyl palmitate alone or with 200 mhl soybean lecithin-isopropyl palmitate gel (wo = 3). No significant alterations of the skin were apparent after 3 days of application of either isopropyl palmitate solvent alone or with the lecithin gel in isopropyl palmitate (Figure 1). In particular, the stratum corneum was still intact after gel treatment. There was no difference between the skin samples treated with gel (Figure 1B) or solvent alone (data not shown) and the control samples treated with physiological NaCl solution (Figure 1C). In all cases, including the control, only an increase in vacuole size of the cells in the spinous layer of the epidermis was observed during the course of the transport experiments. Transdermal Transport of Scopolamine-Scopolamine was used as a reference substance, because its transport through skin and skin models has been studied to a great extent.Sl1 A transdermal device for scopolamine has been commercialized (developed by Aha and marketed by CibaGeigy under the name Transderm-Scop or Scopoderm 'ITS). With this device, a small dose applied behind the ear permits the transdermal transport of scopolamine with beneficial effects against motion-induced sickness.8 Four transdermal transport curves were obtained from scopolamine with four differentmatrices in the donor compartment (Figure 2). The comparison between an aqueous solution and a lecithin gel is shown in Figure 2A, and the transport rate for two different gels, an isopropyl palmitate microemulsion solution (the lecithin organic solution at w, = 0; i.e., priorto gelation)and commercial scopolamine plaster (Scopoderm ?Ts from CibaGeigy) are shown in Figure 2B. The comparison in Figure 2B is made only for comparison with a commercial preparation with regard to the flux intensity. It is not meant to be a comparison of therapeutic efficiency (for which many other parameters should be taken into account, which is not the purpose of the present paper). The results in Figure 2B do, however, show that

"

the lecithin gels have the potential of a rather high transdermal

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was proportional to the concentration (inset of Figure 3). Comparing the microemulsions to a clearly more viscous system (asobtained at w, = 3) shows that the transport rate does not change significantly (Figure 4). Although cyclooctaneis not interesting for physiological application, it is a solvent that permits one to study the dependence on w, in a much broader range (Figure 4B). In fact, gels were obtained at w, = 12 in this case. Again, the results in Figure 4B indicate that the differences between the transport rates of the microemulsionand the gels are within experimental errors. Comparing Figures 4A and 4B shows that the efficiency of transdermal transport is lower for cyclooctane; that is, the influence of the solvent on the transport rate may be significant.

Conclusions
Lecithin organogels can be prepared easily and rapidly and can be obtained with biocompatible components. They are stable for a long time; can incorporate sizeable amounts of quite different chemicals as guest molecules; and seem, therefore, to fulfill the conditions necessary for cosmetic and pharmacological applications. These gels also are transparent, a property allowing the use of spectroscopic methods to detect possible structural changes of the guest molecules and, possibly, the kinetic mechanism involved. In addition to scopolamine and broxaterol reported here, preliminary studies in our laboratory show that various other substances, such as estradiol, amino acids, and peptides, can be transported transdermally via lecithin gels. These results

suggest that there are no great restrictions on the chemical structure of the drug. More studies should be carried out to clarify whether and to what extent the chemical structure really affects the transport rate. The hydrophobicity, the molecular weight, and the charge density are the obvious parameters to investigate first. The gel components are also important, and systematic studies on the effect of the solvent, the concentration of lecithin, and the amount of water are probably also important (water as gel inducer can be substituted for other substances, such as glycerol and other low-molecular-weight, hydrogenbonding liquids). Only preliminary speculations can be offered a t this point on the mechanism of the transport. Perhaps, lecithin gels slightly disorganize the structure ofthe skin, and thus, permit the permeation of various substances. The stratum corneum contains regularly arranged layers of lipids (see classical studies by Elias and Brown=). It is possible that the proposed disorganization is due to interaction between these lipids and the phospholipids of the gel.

References and Notes

1. Scartazzini, R.; Luisi, P. L. J. Phys. Chem. 1988,92,829-833. 2. Luisi, P. L.; Scartazzini, R.; Haering, G.; Schurtenberger, P. Colloid Polym. Sci. 1990,268, 356-374. 3. Schurtenber er, P.; Scartazzini, R.; Ma d, L. J.; Leser, M. E.; Luiei, P. L. Phys. Chem. 1990,94,36&i-3701. 4. Schurtenberger, P.; Scartazzini, R.; Luisi, P. L. Rheol. Acta 1989, 28,372-381. 5. Ott, A.; Urbach, W.; Lan evin, D.; Schurtenberger, P.; ScarPhys.: Condens. Matter 1990, 2, tazzini, R.; Luisi, P. L. 5907-5912. 377-380. 6. Scartazzini, R.;Luisi P. L. Biocatalysis 1990,3, 7. Nastruzzi, C.; Colombo, L.; Willimann, H.; Luisi, P. L., unpublished results. 8. Transdermal Controlled S stemic Medications; Drugs and the Pharmaceutical Sciences; Ciien, Y. W., Ed.; Marcel Dekker: New York, 1987;Vol. 31, pp 1-22. 9. Dr Delivery Systems: Fundamentals and Techniques; Johnson, P.;U&loyd4ones, J. G., Eds.; Ellis Honvood: Chichester, U.K., 1987;pp 200-223. 10. Dermal and Transdermal Absorption; Brandau, R.; Lippold, B. H., Eds.; Wissenschaftliche Verlagsgesellschaft mbH: Stuttgart, Germany, 1982;pp 15&170. 11. Tmnsdermal Delivery of Drugs;K donieus, A. F.; Berner, B., Eds.; CRC: Boca Raton, FL, 1987;$01. I, pp 101-116. 12. Willimann, H.; Luisi, P. L. Biochem. Biophys. Res. Commun. 1991,177, 897-900. 13. Chiarino, D.; Fantucci, M.; Carenzi, A.; Della Bella, D.; Frigeni, V.; Sale R. I1 Farmaco-Ed. Sc. 1986,41,440-453. 14. Chetta, A.; Garavaldi, G.; Cuomo, A.; Gurrieri, G.; Olivieri, D. Respimtion 1988,53,220-224. 15. Rampulla, C.; Corsico, R.; Majani, U.; Lodola, E. Respiration 1985,47,299-302. 16. Robuschi, M.; Simone, P.; Vaghi, A.; Ventresca, G. P.; Bianco, S. Znt. J . Clin. Pharm. Ther. Toxicol. 1987,25,101-104. 17. Simone, P.; Bor ia, M.; Torre, L.; Ventresca, G. P. Eur. J. Clin. Pharmacol. 1998,39, 56-68. 18. Robuschi, M.; Vaghi, A., Gambaro, G.; Refini, M.; Bianco, S. Curr. Ther. Res. 1988,43,725-733. 19. Ufdahl, ,C. G.; Sigvaldasson, A.; Skoogh, B.-E.; Svedmyr, N. Resprmtron 1984,46, 104. 20. Chiravalli, E.; Rimoldi, R. Loth Contro la TBC e le Malattie Polmonari Sociali 1986,56,970. 21. Franz, T. J. Cum. Probl. Dermatol. 1978,7, 58-68. 22. Lucas Meyer (Hamburg, Germany), product information. 23. Elias, P. M.; Brown, B. E. J . Invest. Dermatol. 1979,73,339-348.

I
c 0

l0O0i

5001
0 0

1
1 B

20

40

60

80

Time (h)

600 -n
W

T
O
X

450 I

Time (h)
Figure &Transport of broxaterol through human skin in vitro. (A) Soybean lecithin (200mMHsopropyl palmitate, w, = 0 (curve 1 ; 0)or 3 (curve 2;0). with an initial broxaterol concentration of 40 mglmL. (6) Soybean lecithin (200mM)-cyclooctane,w, = 0 (curve3; 0 )or 12 (curve 4;B), with an initial broxaterol concentration of 20 mg/mL. The corresponding flux value for cyclooctaneis 9.5 pg * h- * cm-2,compared with 17.3c(9 h- for isopropyl palmitate (see inset o f Figure 3).

Acknowledgments
We thank Prof. M. Frei (Department of Plastic Surgery, University Hospital, Zurich) for the skin Sam les and Dr. L. Bruckner (Department of Dermatolo University hospital, Zilrich) for her histological analysis of the%in preparations and for her general interest in the work. We are also grateful to Prof. H. P. Merkle (Department of Pharmacology, ETH-Zentrum, Zilrich) for his comments and suggestions.

874 I Journal of Pharmaceutical Sciences Vol. 81, No. 9, September 7992