EP2299975A1

EP2299975A1 - Preparation and iontophorectic device for the transdermal application of active ingredients

Info

Publication number: EP2299975A1
Application number: EP09777019A
Authority: EP
Inventors: Andreas Koch; Christoph Schmitz; Rolf Pracht
Original assignee: LTS Lohmann Therapie Systeme AG
Current assignee: LTS Lohmann Therapie Systeme AG
Priority date: 2008-07-21
Filing date: 2009-07-08
Publication date: 2011-03-30
Also published as: KR20110051185A; JP2011528672A; CN102131497A; BRPI0916345A2; DE102008034098A1; US20110295184A1; WO2010009808A1

Abstract

The present invention relates to an iontophorectic device and a composition comprising active ingredients for the transdermal release of active ingredients, particularly of human insulin. Insulin, a blood sugar-lowering hormone inhibiting glycogen breakdown of the pancreas, can be made available as the most important pharmaceutical for the therapy of diabetes mellitus via the transdermal resorption route.

Description

PREPARATION AND IONTOPHORETIC DEVICE FOR THE TRANSDERMALE

APPLICATION OF ACTIVE SUBSTANCES

Description:

The present invention relates to a preparation and iontophoretic device for the transdermal delivery of active substances, in particular of human insulin.

Insulin, a hypoglycemic, glycogen-degrading hormone of the pancreas, is the most important drug for the treatment of the disease diabetic mellitus, a sugar use disorder due to relative or absolute lack of insulin with concomitant disorder of fat and protein metabolism and damage to the liver, blood vessel and nervous system ,

Due to its chemical nature, insulin is a peptide and can therefore only be administered via parenteral routes in which no proteolytic processes take place, which would inevitably lead to decomposition of the insulin. Administration via the gastrointestinal tract is unsuitable. As a preferred route of administration for insulin, the subcutaneous injection has prevailed since its introduction, also because it is relatively inexpensive and well done by the patient himself. In addition to the risk of infection, there are other problems for patients who suffer from a so-called syringe phobia. Therefore, other parenteral delivery options for insulin have been sought in recent decades. Most recently, in 2003, an inhaled system of insulin for the pulmonary

Resorptionsweg presented, however, this system was taken after its introduction very quickly withdrawn from the market. Another path that has been the subject of much research in recent years is the transdermal route of absorption. Since the insulin molecule can not overcome the skin permeation barrier due to its physicochemical properties, for example at 5700 daltons it has too large a molecular weight, attempts have been made to bring insulin through the skin into the bloodstream by means of liposomal preparations in the form of so-called Transfersomes ^® . [1] Transfersomes ^® are certain lipid vesicles, ie closed hollow spheres in the nanometer range, consisting of one or more lipid bilayers, wherein highly purified lecithin as phospholipid forms the basic building block or the backbone of the spherical membrane. Transfersomes ^® have towards their aufbauver-turned liposomes from a high membrane flexibility and a strong liposome deformability, which apparently allows them to penetrate even into pores that are much smaller than they are. Since both the interiors of the spherical Transfersomes ^® and Transfersomes ^® as a kind of "shuttle" an interesting galenic dosage form for the dermal or transdermal transport route represent the interiors of the membranes themselves can be loaded with drugs.

Whether liposomes or Transfersomes ^® actually lead to an increased dermal active ingredient penetration compared to conventional galenic preparations, but is in the literature very controversial and is also partly proven by contrary examples. For example, the authors showed in [2] that large hydrophilic molecules, such as the protein insulin, growth hormone or cyclosporin, can not penetrate into the skin via Transfersomes ^® . Own in vitro permeation studies with insulin encapsulated in a transferoma-like preparation failed to provide evidence for a shuttle effect; neither in the acceptor part under the skin (Franz diffusion cell) nor in the skin itself could measurable traces of insulin (mass spectroscopy) be found. The statement of the authors in [1] may also be doubted because a humane clinical study with liposomally prepared insulin for transdermal application, carried out by the applicant, brought a negative result to light. [3]

It was an object of the present invention to overcome the disadvantages of the prior art and to provide peptides or proteins, in particular large hydrophilic molecules, such as growth hormones or cyclosporin and in particular the protein insulin, effectively, but also cost-effectively, suitable for transdermal administration put. The object is achieved by a kit comprising an iontophoresis device and an active ingredient-containing preparation, wherein the active-ingredient-containing preparation contains closed hollow bodies which are present as liposomes or micelles and contain an active substance from the group of peptides or proteins.

All the more surprisingly it could be shown in the present invention that, for example, liposomally encapsulated insulin can be made available in a preparation of the transdermal absorption route by penetrating the liposomally encapsulated insulin into the skin via an "electric drive", for example by iontophoresis. permeates through the skin, although iontophoresis is not applicable as the most effective method of transdermal permeation enhancement of charged drugs, especially for insulin. [4]

The isoelectric point (isoelectric point = pH at which charge balance exists, i.e. the concentration of the anion is equal to that of the cation of an ampholyte) of insulin is at a pH of 5.4; i.e. at a pH <5.4, the insulin molecule carries a positive charge (= cation), while at a pH> 5.4 it is negative (= anion). Since there is a pH gradient in the skin, there is a pH of 5.2 on the surface of the skin, which gradually approaches the physiological pH of 7.4, the deeper one gets through the skin

In the direction of the body's interior or blood vessels, the insulin molecule would inevitably change the state of charge during a transdermal permeation route. The consequent consequence is a transport stop of the transdermal passage; the insulin would even migrate back towards the skin side applied drug part.

Assuming that the pH in insulin was set to 4.0, insulin would be present as a cation and could first penetrate into the skin, provided that the active electrode above the insulin is connected as a positive electrode (= anode).

Due to the pH gradient in the skin, the insulin molecule is initially neutral, the isoelectric point is indeed at a pH of 5.4, and then even increasingly negatively charged. As a result, the electromotive driving force of the iontophoresis in the sense of permeation through the skin no longer work; in the On the contrary, according to the laws of electromigration, the now negatively charged insulin would have to migrate back towards the positive electrode and thus to the drug reservoir.

In the opposite case, i. the active electrode above the insulin would be switched as a negative electrode (= cathode) and the pH of the insulin preparation is equal to or slightly> 7 (from a pH of> 8.5 skin irritation occurs), so that insulin would be negatively charged, the migration or transport conditions are similar; The insulin molecule would already be in the outermost layer of skin, the stratum corneum or cornea, as a neutral molecule and can not continue to penetrate. For substance transport by means of iontophoresis, it is essential that the active substances to be transported are loaded. In the case of peptides, only those suitable as transdermal candidates for iontophoresis are those whose isoelectric point is at a pH <4.0 or> 8.0. [4]

In the case of the present invention, this problem is overcome by the fact that on the one hand, the liposome-encapsulated insulin undergoes no change in its charge conditions during the transdermal penetration route, it is indeed encapsulated and thus electrically shielded and, secondly, probably not the electromigration of the actual transport mechanism Lontophoresis (migration of charged particles to their oppositely charged electrodes) is exploited, but the so-called electroosmosis, a process that arises as a concomitant or by-product when creating an electric field on the skin. Due to the different water content in the individual skin layers, a potential gradient is formed when an electric field is applied, which increases the permeability of the skin and causes the formation of a liquid flow through the skin. This is again responsible for the fact that during iontophoresis even uncharged, electrically neutral molecules, such as liposomal insulin, can increasingly penetrate into the skin [5].

Alone by means of iontophoresis, the transdermal route of insulin or insulin analogues is therefore very difficult and without therapeutic relevance [6] since proteolytic decomposition reactions by proteases occurring in the skin play a major role play. FIG. 2 shows, for example, the degradation of insulin in a human skin sample comminuted by means of Ultra-Turrax treatment under the action of the skin's own proteases. After 8 hours de facto insulin is no longer detectable. It is therefore essential to protect insulin during its transdermal absorption route in particular against the attack of peptide-cleaving proteases. Liposomes are spherical or ovoid structures of one or more concentric lipid bilayers with an aqueous interior, so-called lipid vesicles.

The shape of the liposomes is ultimately irrelevant to the subject invention and may differ significantly from the spherical shape. For the purposes of the present invention, liposomes and micelles are understood as hollow bodies which are able to penetrate into the skin by means of iontophoresis or to permeate through the skin. The diameter of the hollow body is usually in the range of 25 nm to 1 micron. Due to their stability, those with a diameter of 100 to 300 nm are preferably used.

Within the meaning of this invention, the Transfersomes® mentioned in the introduction are likewise suitable as a special active substance-containing preparation for transdermal administration and are included in the application under the term liposomes. For example, the preparation of Transfersomen® in DE 44 47 287 C1, whose disclosure content is an integral part of this description. Usually, liposomes are generated by suspending suitable lipids in aqueous solution. For example, phosphatidylcholines (lecithins), phosphatidylethanolamines or phosphatidylserines (cephalins) are used for this purpose. By treating this mixture with ultrasound, a dispersion of approximately equal closed liposomes is formed. Such liposomes can also be produced, for example, by rapidly mixing an ethanol / lipid solution with water. If the lipid is injected through a thin needle into the aqueous solution, round liposomes of about 50 nm diameter are formed.

Another method (film method) consists of generating a transparent on the inner wall of a round bottom flask by means of rotary evaporator, transparent lipid film. After dissolution of the film in suitable buffer solution become spontaneous produced multilamellar liposomes of different sizes. If the resulting multilamellar liposomal dispersion is extruded several times under pressure through, for example, a polycarbonate membrane of defined pore size, unipolar or oligolaminular liposomes characterized by a homogeneous size distribution are obtained. In principle, these methods are known to the person skilled in the art.

Furthermore, it has surprisingly been found that insulin or Insulinalanolga also the transdermal absorption route can be made accessible via iontophoresis by encapsulating it by micelle formation with ionic or nonionic surfactants. Micelles are the arrangement of individual molecules into a larger association of mostly colloidal order of magnitude (association colloids) with ordered structure due to intermolecular forces.

As micelle-forming molecules are particularly surfactant molecules such as fatty acids, bile acids, alkyl sulfates, fatty alcohol sulfates, fatty alcohol ethersulfate, sulfosuccinates, α-olefinsulfonates, isethionates, alkane and alkylbenzenesulfonates, saponins, quaternary ammonium salts, more preferably sodium dodecyl sulfate or sodium cholate.

Suitable examples of nonionic surfactants are polyethylene glycol ethers, phenol ethoxylates and alkylolamides, particularly preferably octylphenolpoly (ethylene glycol ether) io. Micelle-forming molecules are characterized by containing a hydrophobic hydrocarbon and a hydrophilic group in the molecule. In the case of the formation of spherical or rod-shaped micelles, other molecules can be incorporated in the interior of the associate formed as a result of selective adsorption, which then wetted as in the example of soaps, as oil droplets (lipophilic phase) for water (hydrophilic phase) and thus miscible in water can be.

For effective micelle formation, the selected surfactants must be used at least in amounts corresponding to their so-called critical micelle formation. In the case of insulin or Insulinalanolga once the charges are masked in the molecule, so that for iontophoretic transport processes On the other hand, the insulin molecule is protected by the outer protective sheath of the surrounding surfactant molecules from proteolytic degradation by the skin's own proteases.

The active ingredient-containing preparation is present, for example, as a solution, ointment, paste, foam or gel. In addition to the active substance-containing hollow bodies, it may also contain further auxiliaries, such as lipids, water, alcohols, gel formers, emulsifiers, stabilizers and enhancers. The preparation may also contain portions of active ingredient-containing hollow body with various active ingredients. Thus, for example, short-acting insulin or insulin analogues and insulin or insulin analogues can be administered with long-term effect in one application.

It is only through the combination of iontophoresis and liposomal and / or micelle-forming galenics that peptide (s) or protein (s), which display the hormone- or hormone-like action, in particular insulin or insulin analogues, can be made available transdermally for a sufficiently therapeutic application , According to the invention, at least 10 international units or at least 350 μg of insulin are preferably bioavailable. The content is determined either in vitro via the residual content (HPLC) in the skin after permeation end or directly in vivo via plasma determination or determination of the blood sugar level. The preparation ideally still has at least 50% of its initial active ingredient content during its transdermal passage over a period of 5 hours. In this case, the content is determined in vitro again via the residual content (HPLC) in the skin after previous extraction, in vitro as a model for in vivo.

In addition to the transdermal administration of insulin or insulin analogues, the invention is also suitable for administering physiologically highly active peptides which exhibit hormone- or hormone-like action (peptide hormones), including their derivatives or conjugates having an average molecular weight Mw of 300 to 1,000,000 (Dalton). In general, the peptide hormones are oligo-, more often still to polypeptides (up to 100 amino acids), sometimes to higher molecular weight proteins (proteohormones). The peptide and protein drugs can be used as free acids or bases.

The active ingredient may have an average molecular weight of less than 3,000 Da. Examples of such agents are, in particular, Abarelix, angiotensin II, anidulafungin, antide, argipressin, azalin and azalin B, bombesin antagonist, bradykinin, buserelin, cetrorelix, cyclosporin A, desmopressin, detirelix, enkephaline (Leu, Met) ganirelix, gonadorelin , Goserelin, growth hormone secretagogue, micafungin, nafarelin, leuprolide, leuprorelin, octreotide, orntide, oxytocin, ramorelix, secretin, somatotropin, terlipressin, tetracosactide, teverelix, triptorelin, thyroliberine, thyrotropin, vasopressin.

The active ingredient may have an average molecular weight of 3,000 to 10,000 Da. Examples of such active ingredients are, in particular, calcitonin, corticotropin, endorphins, epithelial growth factor, glucagon, insulin, novoline, parathyroid hormone, relaxin, pro-somatostatin, salmon secretin.

The active ingredient may have an average molecular weight of more than 10,000. Examples of such agents are in particular interferons (alpha, beta, gamma), interleukins (IL1, IL2), somatotropin, erytropoietin, tumor necrosis factor (TNF alpha, beta), relaxin, endorphin, Dornase alpha, folic stimulating hormone (FSH), human chorion gonadotropin (HCG), Human Growth Hormone Release Factor (hGRF), Luteinizing Hormone (LH) or Epidermal Growth Factor.

In the following, the invention will be further explained with reference to Table 1, Figures 1-4 and Examples 0-4.

Table 1 summarizes all in vitro permeation results. The in vitro permeation measurements of the systems according to the invention were carried out on the in vitro skin model of human full skin (FIGS. 1a and 1b) with the aid of modified Franz diffusion cells, equipped as an embodiment for iontophoresis. As acceptor in all cases 0.025 molar HEPES buffer solution was used (HEPES: 2- [4- (2-hydroxyethyl) -1-piperazinyl] ethane sulfonic acid) adjusted to a pH of 7.4, and thermostated at 32 ⁰ C. FIG. 1a represents the in vitro experimental arrangement for the transdermal application of human insulin by combined use of iontophoresis and liposomal or micellar galenics. FIG. 1b illustrates the effect of electroosmosis produced during use.

In it are:

(1) Ag / AgCl electrode as anode and cathode (active electrode and counter electrode). (2) Verum compartment anode or cathode, present as a liposome or as a micelle.

(3) electrolyte compartment cathode or anode, drug-free.

(4) Electrolyte layer, as a conductivity improver for the verum part in the anode or cathode. (5) Adhesive adhesive ring with application fleece for drug absorption.

(6) Upper part of human skin (epidermis with non-pre-damaged stratum corneum)

(7) Liposomes or micelles, electrically neutral or electrically negatively charged, containing human insulin. (8) By electroosmosis, a side effect of iontophoresis, created liquid flow, with the help of which only the insulin-containing liposomes can migrate or permeiern.

(9) dermis with blood capillaries for the systemic removal of the transdermally applied active substance.

As a current source, a DC generator was usually used (Hameg HM 7042-5, Hameg from Mainhausen, Germany), which was set to a constant current output of 500 uA / cm ² absorption surface skin. Full-area Ag / AgCl electrodes from NAIMco, Chattanooga (USA) were used as the electrode material. The electrolyte reservoir of the counterelectrode (cathode or also anode) consisted of a 2% solution of hydroxypropylcellulose with 0.9% by weight addition of NaCl, which had a weight per unit area of 3 g / 30 cm ² on a nonwoven consisting of nonwoven polyester fleece (Paramol N 260-300P, Fa. Lohmann & Rauscher, Neuwied, Germany). In the anode or Kathodenreservoir, the equipment as an anode or cathode was directed to the charge of the corresponding hollow body, about 200 mg of the corresponding insulin preparation were applied to the identical web of the counter electrode. For the purpose of fixation on the skin, the edge consisted of a self-adhesive polyolefin foam ring (3M, type 1779). The duration of permeation or iontophoresis

Treatment was 5 hours for each of the experiments; in the case of the Transfersomes, the electrodes were each reversed after 2.5 h. The contact between the cells is made by a silver wire as a bridge.

After Permeationsende the skin was subjected to a residual content determination on insulin, in which it was first cut with a pair of scissors into several small parts, then extracted for 5 hours with shaking in 70% hydrochloric acid ethanol, followed by a specific HPLC method for insulin to be examined. In order to show that insulin has indeed overcome the main permeation barrier of the human skin, the stratum corneum, the skin has been prepared for residual content determination in such a way that the stratum corneum is stimulated by means of so-called "tape stripping" [7] In order to further emphasize the selectivity of the analytical procedure used and the degree of truth of the insulin detected, the in vitro permeation experiments were carried out in each case with verum and placebo samples (same galenic preparation without active ingredient).

FIG. 2 shows the proteolytic degradation process of isolated insulin, ie the insulin is neither liposomally nor micellar encapsulated and in comparison thereto the markedly lower degradation of insulin which is protected from proteases by liposome or micelle formation. FIGS. 3 and 4 show, by way of example, HPLC chromatograms of verum and placebo samples from the permeation studies for the insulin preparations, here for inventive example 1. In the placebo sample, no peak is detectable in the retention time range of insulin; Thus, the peak recognized as insulin is actually insulin, as evidenced by the chromatograms in Figure 4 (HPLC chromatogram of the corresponding standard insulin sample) and UV spectral comparison standard and verum sample (not shown here as a figure) becomes. The same could be shown for the other examples of the invention, therefore only once exemplified.

FIG. 3 shows insulin-liposome chromatograms (emission) of residual contents of human whole skin dissolved in 70%, 0.01 m hydrochloric acid ethanol, where verum (1) and placebo (2) are shown.

Figure 4 shows insulin standard and insulin liposome chromatograms (emission) residual contents of human whole skin dissolved in 70%, 0.01 M hydrochloric acid ethanol, with insulin standard dilution SV3, c = 11, 01 ug / ml (1) and Verum (2) is shown.

Table 1: Outcome summary Insulin iontophoresis experiments with various galenical preparations, each masking or masking the intrinsic charge of the insulin

I ¹ - Switching the verum compartment as a cathode, as the vehicle carries a negative charge to the outside.

I ² - switching of the verum compartment first as a cathode, then after 2.5 h reversing as the anode, since the vehicle is electro-neutral to the outside.

The results show that it is possible to carry out a transdermal therapy for diabetes mellitus type 1 with acceptable TTS sizes <-100 cm ² , at least for inventive examples 1-3. In particular, for the treatment of a chronic metabolic disorder (diabetes mellitus), which is based on a lack of insulin or reduced insulin action, can be treated excellently with the article according to the invention.

For transdermal administration of insulin or insulin analogs is described by the following

Process steps characterized: a) Preparation of a preparation containing active ingredient containing insulin or

Insulin analogues in closed hollow bodies, wherein the hollow bodies are present as liposomes or micelles. b) applying the preparation together with the iontophoresis device to the skin c) carrying out the iontophoresis

FIG. 2 shows the proteolysis of insulin as a function of the contact time with in vitro human skin material. The effect of skin-specific proteases on the degradation of insulin after a defined-content insulin buffer solution together with in vitro human skin punches (24 cm ² ) crushed by means of an Ultra-Turrax disperser, over a period of 8 hours Stirring were in contact, can be clearly seen. The reduction in the micellar insulin sample (Triton-X 100 ^® as a micelle-forming non-ionic surfactant) extends significantly slowed; here the insulin content is still well over 50% even after 8 hours of contact time. In this mean:

A Reference solution B, C Insulin unencapsulated D Insulin micellar encapsulated

As a technical embodiment in practice, in principle, as described above for the in vitro permeation or absorption studies, the same system can be used. As electrode material, for example, full-surface Ag / AgCl electrodes from NAImco, Chattanooga (USA) are used, which are available in different sizes (1, 5 - 4 cm ² ), are self-adhesive and equipped with a polyester fleece for holding the active ingredient preparations. The active ingredient-containing preparation is housed either in the anodic or cathodic part of the iontophoresis device.

The electrolyte reservoir of the counterelectrode (cathode or anode), for example, again consists of a 2% solution of hydroxypropylcellulose with 0.9 wt .-% addition of sodium chloride, with a basis weight corresponding to 3 g / 30 cm ² in or on the Application fleece of Naimco ^® electrodes applied. As counterelectrodes, for example, the corresponding electrodes ["Dispersive (Return) electrode] of the iontophoresis application kit (" ionto + plus ") can also be used. The permeation time or iontophoresis treatment should preferably not exceed a period of 5 hours, preferably after half the duration of the iontophoresis the electrodes are reversed polarity when externally electrically neutral hollow bodies are used, in the case of using liposomes and micelles with nonionic surfactants, the electrodes are reversed after 2.5 hours each after 5 hours of use.

As a power source with regulation or control of the current strength, a corresponding device are also used by the company NAImco. Type "id ³ drug delivery device", which due to its size is fastened to the upper arm or above the wrist by means of Velcro straps, for example.The electrodes are connected to the power generator via conventional cables with banana plug connection.

Inventive Example 1: Preparation of a micellar preparation by means of sodium oxycholate

First, a Tris / HCl buffer / glycerol / water mixture (pH 6.8) having the following composition is prepared (tris: 2-amino-2- (hydroxymethyl) -propane-1,3-diol or amino-tris ( hydroxymethyl) methane). 25.0 ml 0.5 M Tris / 1 M HCl 11, 5 mL Glycerol 63.5 mL Water

366 mg of sodium oxycholate, corresponding to 8.5 mmol, are then introduced into this mixture and stirred until a clear solution has formed (solution A). Subsequently, approx. 100 mg of human insulin are weighed into a 10 ml volumetric flask and filled up with solution A. The batch is stirred until complete dissolution of the insulin or for at least one hour at room temperature and then used directly for the Permeationsexperimente.

Inventive Example 2 Preparation of a micellar preparation by means of sodium dodecylsulfate The preparation is analogous to Invention Example 1 with the difference that instead of sodium oxycholate 2 g of sodium dodecyl sulfate are added. The batch is again stirred until complete dissolution of the insulin or for at least one hour at room temperature and then used directly for the Permeationsexperimente.

Invention Example 3: Preparation of a micellar preparation by Triton X-100 ^® is prepared analogously to the invention example 1 with the difference that instead of sodium Oxycholat 323.5 mg Triton X-100 ^® (corresponding to 5 mmol or 0.3%) are introduced , where only the Triton X-100 ^® must be submitted, then it is filled with the buffer glycerol mixture.

Furthermore, are weighed in place of 100 mg of human insulin only 15.4 mg in a 100 ml volumetric flask and X-100 ^® -containing buffer-glycerol mixture with the Triton (Triton X-100 ^® = octylphenolpoly (ethylene glycol ether) -ιo) refilled. The batch is again stirred until complete dissolution of the insulin or for at least one hour at room temperature and then used directly for the Permeationsexperimente.

Invention Example 4 Production of a liposomal preparation by means of L-α-phosphatidylcholine by the film method [8]

First, 187.2 mg of L-α-phosphatidylcholine (lecithin) are dissolved in 10 ml of methanol. 2.5 ml of this lipid stock solution are then transferred by means of an Eppendorf pipette into a 50 ml round bottom flask, the tip is then rinsed twice with 2.5 ml methanol. By stripping off the methanol on a rotary evaporator

(40 ⁰ C, rotation step 4-5, it may no longer be perceptible odor methanol) A transparent lipid film which is in high vacuum (p = 0.05 mbar) post-dried for 2 hours, to remove any solvent residues. 2 ml of a 0.1% insulin-containing buffer solution, which is composed as follows, are then pipetted into the round-bottomed flask.

10 mmol of HEPES and 150 mmol of NaCl adjusted to pH 7.4 with 5 N NaOH. Human insulin is dissolved in 5 ml of this buffer solution with stirring for one hour. The round bottom flask containing the insulin-containing buffer solution is shaken overnight at 240 revolutions per minute on a shaker. Comparative Example 0: Preparation of an insulin-containing buffer solution without micelle and / or liposome formation

The preparation is analogous to Invention Example 1 with the difference that no sodium oxycholate is entered. The mixture is stirred again until complete dissolution of the insulin or for at least one hour at room temperature.

From all batches (Comparative Example 0 and Inventive Examples 1-4), corresponding preparations without human insulin (placebo batches) were prepared, which were tested together with the respective verum batches in the iontophoretic in vitro permeation experiments (as insulin negative control or proof of the selectivity of the analytical HPLC method used for determining the residual content of insulin in the skin).

reference

[1] G. Cevc / Drug Delivery across the skin. Exp. Opin. Invest. Drugs 6, p. 1887-

1937 (1997) [2] H. Schreier, J. Bouwstra / Liposomes and niosomes as topical drug carriers: dermal and transdermal drug delivery. J. Contr. Rel. 30, pp. 1-155 (1994) [3] LTS Clinic Study Eutra CT No .: 2004-004043-21

[4] Y.N. Kalia et al. / Advanced Drug Delivery Reviews 56 (2004) 619-658 [5] R.R.Burnette / Lontophoresis, in: Transdermal Drug Delivery - Develop-mentally

Issues and Research Initiatives, (Eds.) J. Hadgraft and R.H. Guy, Marcel Dekker

New York, 247-292 (1989) [6] L. Langkjaar et al. / Lontophoresis of monomeric insulin analogues in vitro:

Effects of insulin charge across skin pre-treatment. J. controlled release 51: p.

42-56 (1998) [7] C. Herkenne et al. / In vivo Methods continue Assessment of Topical Drug

Bioavailability: Pharm. Res., Vol. 25, no. 1, pp. 87-103 (2008) [8] A. Bangham / Diffusion of Univalent ions across the lamellae of swollen phospholipids: J Mol Biol 13 (1): 238-252 (1965)

Claims

claims

A kit comprising an iontophoresis device and an active ingredient-containing preparation, wherein the active-substance-containing preparation contains closed hollow bodies which are present as liposomes or micelles and contain an active substance from the group of peptides or proteins.

2. Kit according to claim 1, wherein the active ingredient-containing preparation is present as a solution, ointment, paste, foam or gel.

3. Kit according to claim 1 or 2, wherein the active ingredient in the preparation is a peptide hormone or a proteo-hormone, in particular insulin or an insulin analogues.

A kit according to any one of the preceding claims, wherein at least 10 international units or at least 350 μg of insulin are bioavailable.

5. Kit according to one of the preceding claims, wherein the content of active ingredients) in the preparation during its transdermal passage in a period of 5 hours still at least 50% of its initial content.

6. Kit according to any one of the preceding claims, wherein the active ingredient-containing preparation is housed in the anodic or cathodic part of the iontophoresis device.

7. Kit according to one of the preceding claims, wherein after half the duration of the iontophoresis, the electrodes are reversed.

8. Kit according to any one of the preceding claims, wherein the active electrode and counter electrode each consist of films which are coated with a mixture of silver and silver chloride.

9. Kit according to one of the preceding claims, wherein both the active electrode and the counter electrode are designed as self-adhesive systems.

10. Kit according to claim 1 to 9, wherein the micelles are composed of ionic surfactants.

The kit of claim 10, wherein the ionic surfactant is selected from the group consisting of fatty acids, bile acids, alkyl sulfates, fatty alcohol sulfates, fatty alcohol ether sulfates, sulfosuccinates, α-olefin sulfonates, isethionates, alkane and alkyl benzene sulfonates and saponins.

The kit of claim 11, wherein the ionic surfactant is sodium dodecyl sulfate or sodium cholate.

13. Kit according to claim 1 to 9, wherein the micelles are composed of nonionic surfactants.

14. Kit according to any one of the preceding claims 13, wherein the nonionic surfactant used is selected from the group comprising polyethylene glycol ethers, phenol ethoxylates and alkylolamides.

15. Kit according to any one of the preceding claims 14, wherein the nonionic surfactant is octylphenolpoly (ethylene glycol ether) io.

16. Kit according to claim 1 to 9, wherein the liposomes are composed of lipids selected from the group comprising phosphatidylcholine, phosphatidylethanolamine or phosphatidylserine.

17. A method for the transdermal administration of insulin or insulin analogues by the following steps: a. Preparation of a preparation containing active ingredient containing insulin or insulin analogues in closed hollow bodies, wherein the hollow bodies are present as liposomes or micelles. b. Applying the preparation together with the iontophoresis device to the skin c. Carrying out the iontophoresis

18. A method for the transdermal administration of insulin or insulin analogs according to claim 17, wherein after half the duration of the iontophoresis the electrodes are reversed.

19. Use of Kit claims 1 to 16 for the treatment of a chronic metabolic disorder (diabetes mellitus), which is based on a lack of insulin or reduced insulin action.