Classifications

• • A—HUMAN NECESSITIES
  • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
  • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
  • A61K9/00—Medicinal preparations characterised by special physical form
  • A61K9/0012—Galenical forms characterised by the site of application
  • A61K9/0014—Skin, i.e. galenical aspects of topical compositions
• • A—HUMAN NECESSITIES
  • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
  • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
  • A61K31/00—Medicinal preparations containing organic active ingredients
  • A61K31/185—Acids; Anhydrides, halides or salts thereof, e.g. sulfur acids, imidic, hydrazonic or hydroximic acids
  • A61K31/19—Carboxylic acids, e.g. valproic acid
  • A61K31/192—Carboxylic acids, e.g. valproic acid having aromatic groups, e.g. sulindac, 2-aryl-propionic acids, ethacrynic acid 
• • A—HUMAN NECESSITIES
  • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
  • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
  • A61K31/00—Medicinal preparations containing organic active ingredients
  • A61K31/33—Heterocyclic compounds
  • A61K31/395—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins
  • A61K31/40—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having five-membered rings with one nitrogen as the only ring hetero atom, e.g. sulpiride, succinimide, tolmetin, buflomedil
  • A61K31/403—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having five-membered rings with one nitrogen as the only ring hetero atom, e.g. sulpiride, succinimide, tolmetin, buflomedil condensed with carbocyclic rings, e.g. carbazole
  • A61K31/404—Indoles, e.g. pindolol
  • A61K31/405—Indole-alkanecarboxylic acids; Derivatives thereof, e.g. tryptophan, indomethacin
• • A—HUMAN NECESSITIES
  • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
  • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
  • A61K31/00—Medicinal preparations containing organic active ingredients
  • A61K31/33—Heterocyclic compounds
  • A61K31/395—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins
  • A61K31/54—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having six-membered rings with at least one nitrogen and one sulfur as the ring hetero atoms, e.g. sulthiame
• • A—HUMAN NECESSITIES
  • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
  • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
  • A61K31/00—Medicinal preparations containing organic active ingredients
  • A61K31/56—Compounds containing cyclopenta[a]hydrophenanthrene ring systems; Derivatives thereof, e.g. steroids
  • A61K31/57—Compounds containing cyclopenta[a]hydrophenanthrene ring systems; Derivatives thereof, e.g. steroids substituted in position 17 beta by a chain of two carbon atoms, e.g. pregnane or progesterone
  • A61K31/573—Compounds containing cyclopenta[a]hydrophenanthrene ring systems; Derivatives thereof, e.g. steroids substituted in position 17 beta by a chain of two carbon atoms, e.g. pregnane or progesterone substituted in position 21, e.g. cortisone, dexamethasone, prednisone or aldosterone
• • A—HUMAN NECESSITIES
  • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
  • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
  • A61K31/00—Medicinal preparations containing organic active ingredients
  • A61K31/59—Compounds containing 9, 10- seco- cyclopenta[a]hydrophenanthrene ring systems
• • A—HUMAN NECESSITIES
  • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
  • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
  • A61K45/00—Medicinal preparations containing active ingredients not provided for in groups A61K31/00 - A61K41/00
  • A61K45/06—Mixtures of active ingredients without chemical characterisation, e.g. antiphlogistics and cardiaca
• • A—HUMAN NECESSITIES
  • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
  • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
  • A61K9/00—Medicinal preparations characterised by special physical form
  • A61K9/10—Dispersions; Emulsions
  • A61K9/107—Emulsions ; Emulsion preconcentrates; Micelles
  • A61K9/113—Multiple emulsions, e.g. oil-in-water-in-oil
• • A—HUMAN NECESSITIES
  • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
  • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
  • A61P17/00—Drugs for dermatological disorders
• • A—HUMAN NECESSITIES
  • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
  • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
  • A61P17/00—Drugs for dermatological disorders
  • A61P17/02—Drugs for dermatological disorders for treating wounds, ulcers, burns, scars, keloids, or the like
• • A—HUMAN NECESSITIES
  • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
  • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
  • A61P17/00—Drugs for dermatological disorders
  • A61P17/06—Antipsoriatics
• • A—HUMAN NECESSITIES
  • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
  • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
  • A61P17/00—Drugs for dermatological disorders
  • A61P17/08—Antiseborrheics
• • A—HUMAN NECESSITIES
  • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
  • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
  • A61P43/00—Drugs for specific purposes, not provided for in groups A61P1/00-A61P41/00

Definitions

• the present invention relates to a topical pharmaceutical composition for cutaneous application comprising a pharmacologically active agent, a surfactant, a co-solvent and an aqueous phase.
• Psoriasis is a chronic inflammatory skin disease that manifests as erythematous, dry, scaling plaques resulting from hyperkeratosis.
• the plaques are most often found on the elbows, knees and scalp, though more extensive lesions may appear on other parts of the body, notably the lumbosacral region.
• the most common treatment of mild to moderate psoriasis involves topical application of a composition containing a corticosteroid as the active ingredient. While efficacious, corticosteroids have the disadvantage of a number of adverse effects such as skin atrophy, striae, acneiform eruptions, perioral dermatitis, overgrowth of skin fungus and bacteria, hypopigmentation of pigmented skin and rosacea.
• an advantageous non-steroidal treatment of psoriasis has consisted in topical treatment with the vitamin D analogue compound, calcipotriol, formulated in an ointment composition (marketed as Daivonex ® or Dovonex ® ointment by LEO Pharma) in which the calcipotriol is present in solution or a cream composition (marketed as Daivonex ® or Dovonex cream by LEO Pharma).
• the solvent in the ointment composition is propylene glycol which has the advantage of enhancing penetration of the active ingredient into the skin, leading to an improved efficacy, but which is also known to act as a skin irritant.
• Daivonex ointment Due to the improved penetration of calcipotriol into the skin resulting, inter alia, from the presence of propylene glycol, Daivonex ointment has been found to be more efficacious in the treatment of psoriatic lesions than Daivonex ® cream, but has also caused skin irritation in a significant proportion of psoriasis patients. It is therefore an object of the invention to provide a topical composition comprising a vitamin D derivative or analogue as the active ingredient, which has skin penetration and biological activity properties comparable to those of Daivonex ® ointment, but which does not contain propylene glycol as the solvent.
• Human skin in particular the outer layer, the stratum corneum, provides an effective barrier against penetration of microbial pathogens and toxic chemicals. While this property of skin is generally beneficial, it complicates the dermal administration of pharmaceuticals in that a large quantity, if not most, of the active ingredient applied on the skin of a patient suffering from a dermal disease may not penetrate into the viable layers of the skin where it exerts its activity.
• Propylene glycol is a well-known penetration enhancer, i.e. a substance which is capable of penetrating the stratum corneum and "draw" low-molecular components such as therapeutically active components in the vehicle into the epidermis.
• Propylene glycol may in itself give rise to significant skin irritation, and it is also capable of "drawing" low- molecular and potentially irritative components of the vehicle into the epidermis, leading to an overall irritative effect of conventional vehicles including propylene glycol. For this reason, the presence of propylene glycol as a solvent in compositions intended for the treatment of inflammatory skin diseases may exacerbate the inflammatory response.
• compositions of the invention exhibit a comparable or higher biological activity to that of Daivonex ® ointment as determined in the target gene activation assay described in Example 4 below.
• the composition is physically stable, and the vitamin D analogue is chemically stable therein.
• the present invention relates to a topical composition for cutaneous application which is an oil-in-water-in-oil emulsion comprising an aqueous phase containing, dispersed therein, a lipophilic phase comprising
• a non-ionic surfactant selected from the group consisting of polyoxyl glycerides, polyoxyethylene castor oil derivatives, polyoxyethylene alkyl ethers, polysorbates or poloxamers; and
• aqueous phase being dispersed in a pharmaceutically acceptable anhydrous lipophilic carrier or vehicle.
• the invention relates to a topical composition as described herein for use in the prevention or treatment of dermal diseases or conditions.
• Figs. 1 -4 are graphs showing the solubility of calcipotriol monohydrate in co-solvent- surfactant mixtures included in the present composition compared to the solubility of calcipotriol monohydrate in either the co-solvent or the surfactant alone.
• Fig. 5a and 5b are graphs showing the penetration into the skin of composition of the invention.
• Fig. 6 is a schematic representation of the activation of the gene encoding cathelicidin by vitamin D 3 in human keratinocytes. The mechanism of cathelicidin gene activation is used in a biological assay using reconstructed human epidermis (human keratinocytes cultured so as to form the epidermal layers characteristic of human skin) on which calcipotriol-containing compositions of the invention are applied to activate cathelicidin as described in detail in Example 4 below.
• non-ionic surfactant is intended to indicate a surfactant comprising a hydrophilic and a hydrophobic portion in which the hydrophilic portion carries no charge but derives its surface activity from highly polar groups such as polyoxyethylene groups.
• the surfactant is an oil-in-water surfactant with an HLB value of 9- 18.
• lower alkanol co-solvent is intended to indicate a solvent consisting essentially of a Ci -6 straight or branched alkanol, e.g. methanol, ethanol, propanol, isopropanol or butanol.
• vitamin D derivative is intended to indicate a biologically active metabolite of vitamin D 3 , such as calcitriol, or a precursor to such a metabolite, such as alfacalcidol.
• vitamin D analogue is intended to indicate a synthetic compound comprising a vitamin D scaffold with sidechain modifications and/or modifications of the scaffold itself.
• the analogue exhibits a biological activity on the vitamin D receptor comparable to that of naturally occurring vitamin D compounds.
• Calcipotriol has been found to exist in two crystalline forms, an anhydrate and a
• Calcipotriol monohydrate and its preparation are disclosed in WO 94/15912.
• storage stability is intended to indicate that the composition exhibits chemical and physical stability characteristics that permit storage of the composition, at refrigeration or, preferably, room temperature for a sufficient period of time to make the composition commercially viable, such as at least 12 months, in particular at least 18 months, and preferably at least 2 years.
• chemical stability or “chemically stable” is intended to indicate that no more than 10%, preferably no more than 5%, of the vitamin D derivative or analogue degrades over the shelf-life of the product, typically 2 years.
• An approximation of chemical stability at room temperature is obtained by subjecting the composition to accelerated stability studies at 40°C. If less than about 10% of the substance has degraded after 3 months at 40°C, this is usually taken to correspond to a shelf-life of 2 years at room temperature.
• “chemical stability” is intended to mean that the calcipotriol does not degrade significantly over time to 24-epi calcipotriol or other degradation products of calcipotriol in the finished pharmaceutical product.
• composition retains its macroscopic and microscopic appearance over the shelf-life of the product, e.g. that the vitamin D derivative or analogue does not precipitate from the solvent phase or that there is no visible phase separation of the solvent phase and the carrier phase.
• substantially anhydrous is intended to mean that the content of free water in the lipophilic carrier or vehicle does not exceed about 2% by weight, preferably not about 1% by weight, of the carrier or vehicle.
• vitamin D derivative or analogue is intended to indicate the solubility of the vitamin D derivative or analogue, in some instances several fold higher, when a combination of co-solvent and surfactant is present in the aqueous phase than the sum of solubilities in either the co-solvent or the surfactant when these are added individually to the aqueous phase.
• skin penetration is intended to mean the diffusion of the active ingredient into the different layers of the skin, i.e. the stratum corneum, epidermis and dermis.
• skin permeation is intended to mean the flux of the active ingredient through the skin into the systemic circulation or, in case of in vitro studies such as those reported in Example 3 below, the receptor fluid of the Franz cell apparatus used in the experiment.
• biological activity is intended to mean the activity of a vitamin D derivative or analogue when applied to skin in a composition of the invention.
• the biological activity of compositions is determined in an in vitro assay measuring the activation of a target gene encoding cathelicidin in a reconstructed human epidermis model involving cultured human keratinocytes, as described in detail in Example 5 below.
• the composition comprises a vitamin D derivative or analogue selected from the group consisting of calcipotriol, calcitriol, tacalcitol, maxacalcitol, paricalcitol and alfacalcidol.
• the composition comprises calcipotriol or calcipotriol monohydrate as the vitamin D analogue.
• the surfactant is generally present in a concentration of from about 0.5% by weight to about 5% by weight, or from about 1% by weight to about 3% by weight, or from about 1.2% by weight to about 2% by weight, such as about 1.5% by weight, of the composition.
• the non-ionic surfactant is preferably selected from the group consisting of polyethylene glycol 8 caprylic/capric glyceride (a polyethylene glycol derivative of a mixture of mono-, di- and triglycerides of caprylic and capric acids with an average of 8 moles of ethylene oxide) or polyethylene glycol 6 caprylic/capric glyceride (a polyethylene glycol derivative of a mixture of mono-, di- and triglycerides of caprylic and capric acids with an average of 6 moles of ethylene oxide).
• the non-ionic surfactant is favourably polyethylene glycol 8 caprylic/capric glyceride, e.g. available from Gattefosse under the trade name Labrasol or from Condea under the trade name Softigen 767.
• the non-ionic surfactant may also preferably be a polyethylene glycol C 6 - 2 o fatty acid glyceride selected from the group consisting of caprylocaproyl PEG glyceride, lauroyl PEG glyceride, linoeoyl PEG glyceride, oleoyl PEG glyceride and stearoyl PEG glyceride, a polyoxyethylene C 8 .
• alkyl ether selected from the group consisting of PEG monocetyl ether, PEG monolauryl ether, PEG monooleyl ether and PEG monostearyl ether, a polysorbate selected from the group consisting of polysorbate 20, 40, 60 and 80, a poloxamer selected from the group consisting of poloxamer 124, 237, 338 and 407, or a polyoxyethylene castor oil derivative such as polyoxyl castor oil or hydrogenated polyoxyl castor oil.
• the composition further comprises a lower alkanol co-solvent which may favourably be selected from the group consisting of methanol, ethanol, n-propanol, isopropanol, n-butanol or 2-butanol.
• a lower alkanol co-solvent which may favourably be selected from the group consisting of methanol, ethanol, n-propanol, isopropanol, n-butanol or 2-butanol.
• the lower alkanol co- solvent may favourably be present in a concentration of about 0.5-5 %, in particular about 1- 3%, or about 2%, by weight of the composition.
• the co-solvent is ethanol and the non-ionic surfactant is polyethylene glycol 8 caprylic/capric glyceride, polysorbate 80 or PEG monocetyl ether, or the co-solvent is isopropanol and the non-ionic surfactant is polyoxyl castor oil, polyethylene glycol 8 caprylic/capric glyceride or polysorbate 80.
• the ointment carrier may be a hydrocarbon or mixture of hydrocarbons with chain lengths ranging from C 5 to C 6 o.
• a frequently used ointment carrier is petrolatum, or white soft paraffin, which is composed of hydrocarbons of different chain lengths peaking at about C 40- 44, or a mixture of petrolatum and liquid paraffin (consisting of hydrocarbons of different chain lengths peaking at C 28-40 ). While petrolatum provides occlusion of the treated skin surface, reducing transdermal loss of water and potentiating the therapeutic effect of the active ingredient in the composition, it tends to have a greasy and/or tacky feel which persists for quite some time after application, and it is not easily spreadable.
• paraffins consisting of hydrocarbons of a somewhat lower chain length, such as paraffins consisting of hydrocarbons with chain lengths peaking at C 14.16, Ci 8-22 , C 20- 22, C 2 o- 26 or mixtures thereof (the hydrocarbon composition of the paraffins has been determined by gas chromatography). It has been found that such paraffins are more cosmetically acceptable in that they are less tacky and/or greasy on application and more easily spreadable. They are therefore expected to result in improved patient compliance. Suitable paraffins of this type, termed petrolatum jelly, are manufactured by Sonneborn and marketed under the trade name Sonnecone, e.g. Sonnecone CM, Sonnecone DM1 , Sonnecone DM2 and Sonnecone HV. These paraffins are further disclosed and characterized in WO 2008/141078 which is incorporated herein by reference.
• a lipophilic viscosity-increasing ingredient such as a wax.
• the wax may be a mineral wax composed of a mixture of high molecular weight hydrocarbons, e.g. saturated C 3 5 -7 o alkanes, such as microcrystalline wax.
• the wax may be a vegetable or animal wax, e.g. esters of C i4 -32 fatty acids and Ci 4-32 fatty alcohols, such as beeswax.
• the amount of viscosity- increasing ingredient may vary according to the viscosifying power of the ingredient, but may typically be in the range of about 1 -20% by weight of the composition.
• the viscosity-increasing ingredient is microcrystalline wax it is typically present in an amount in the range of about 5-15% by weight, e.g. about 10% by weight, of the composition.
• the composition may additionally comprise an emollient which may act to soften the thickened epidermis of the psoriatic plaques.
• a suitable emollient for inclusion in the present composition may be a silicone wax or a volatile silicone oil as the presence of silicone has additionally been found to aid penetration of calcipotriol into the skin.
• Compositions including a silicone have also been found to result in less skin irritation.
• Suitable silicone oils for inclusion in the present composition may be selected from cyclomethicone, dimethicone.
• the amount of silicone oil included in the present composition is typically in the range of from about 1 to about 10% by weight, e.g. about 5% by weight, of the composition.
• Daivonex ® ointment the presence of propylene glycol is believed to be a major contributor to the skin irritation experienced by many patients.
• calcipotriol may in itself be mildly irritative in some patients (A. Fullerton and J. Serup, Br. J. Dermatol. 137, 1997, pp. 234-240 and A. Fullerton et al., Br. J. Dermatol. 138, 1998, pp. 259-265).
• an anti-irritant compound such as glycerol, butylene glycol, sorbitol, sucrose, saccharin, menthol or nicotinamide.
• Glycerol has been described as a substance that is capable of protecting the skin against irritative substances (J. Bettinger et al., Dermatology 197, 1998, pp. 18-24) and has been found by us to reduce the release of IL-la in a dose-dependent manner: thus, it has been found that the presence of 15% by weight of glycerol in a calcipotriol ointment results in a significantly lower level of release of IL-la than does the inclusion of 10% by weight of glycerol which, in turn, results in a significantly lower level of IL-l release than does the inclusion of 5% by weight of glycerol.
• glycerol is capable of potentiating the biological activity of calcipotriol in that the expression of cathelicidin (in the assay described in Example 4 below) has been found to be increased with decreasing amounts of glycerol in the composition (i.e. more cathelicidin is expressed when the amount of glycerol is 5% by weight than when the amount of glycerol is 10% or 15%, respectively): this implies that with respect to inclusion of glycerol a balance has to be struck between a favourable anti-irritative effect and a favourable potentiating effect.
• the inclusion of about 5-10% by weight of glycerol in the present composition results in a significant anti-irritative effect as well as a significant potentiation of the biological activity of calcipotriol.
• Calcipotriol is known to be a substance which is extremely sensitive to acid conditions (pH below about 7.0 in aqueous compositions or acidic reacting substances in non-aqueous compositions) which contribute to the rapid degradation of calcipotriol.
• a compound capable of neutralizing acidic impurities which may be present in one or more of the excipients of the composition and which are detrimental to the chemical stability of calcipotriol.
• the acid neutralizing compound may favourably be selected from a buffer such as a phosphate buffer which may be included in an amount of about 0.025-0.1% by weight of the composition.
• the acid neutralizing compound may also be an amine such as triethanolamine, trometamol, monoethanolamine or diethanolamine, which may be included in the composition in an amount of about 0.1-2% by weight.
• a water-in-oil emulsifier with an HLB value of 3-8.
• emulsifiers are polyoxyethylene C 8- alkyl ethers, e.g. polyoxyethylene stearyl ether, polyoxyethylene cetyl ether or
• the amount of water in the composition may range from about 1% to about 15% by weight, e.g. from about 5% to about 10% by weight, of the composition.
• the present composition comprises
• the present composition comprises
• composition comprises
• the present composition comprises
• the present composition comprises
• the present composition comprises
• the present composition may also comprise other components commonly used in dermal formulations, e.g. antioxidants (e.g. alpha-tocopherol), preservatives, sodium edetate, pigments, skin soothing agents, skin healing agents and skin conditioning agents such as urea, allantoin or bisabolol, cf. CTFA Cosmetic Ingredients Handbook, 2 nd Ed., 1992.
• antioxidants e.g. alpha-tocopherol
• composition of the invention may be used in the treatment of psoriasis, sebopsoriasis, pustulosis palmoplantaris, dermatitis, ichtyosis, rosacea and acne and related skin diseases by topically administering an effective amount of a composition according to the invention to a patient in need of such treatment.
• Said method preferably comprises topical administration once or twice a day of a therapeutically sufficient dosage of said composition.
• the composition according to the invention preferably contains about 0.001 -0.5 mg/g, preferably about 0.002-0.25 mg/g, in particular 0.005-0.05 mg/g, of the vitamin D derivative or analogue. It is envisaged that the present composition may advantageously been used for maintenance treatment of these dermal diseases, i.e. continued treatment after the
• additional therapeutically active ingredients include, but are not limited to, anti-inflammatory drugs such as corticosteroids, such as betamethasone and esters thereof, e.g.
• non-steroidal antiinflammatory drugs such as naproxen, indomethacin, diclofenac, ibuprofen, dexibuprofen, ketoprofen, flurbiprofen, piroxicam, tenoxicam, lornoxicam or nabumeton, phosphodiesterase 4 inhibitors or p38 MAP kinase inhibitors.
• compositions of the invention are provided.
• compositions shown in Tables la and lb below were prepared by initially mixing the surfactant (Cremophor EL, Labrasol, polysorbate 80 or cetomacrogol 1000) with the co- solvent (ethanol or isopropanol), dissolving calcipotriol monohydrate in the mixture and finally adding the mixture to the aqueous buffer solution adjusted to pH 8.0 and glycerol (when included). The resulting dispersion was then mixed with a mixture of paraffins, emulsifier (polyoxyethylene stearyl ether), DL-a-tocopherol and sodium edetate.
• compositions G-L of the invention are Compositions G-L of the invention.
• compositions M and N of the invention are Compositions M and N of the invention.
• compositions M and N were prepared essentially as described above for compositions A-L with the exception that a mixture of Petrolatum Jelly White (Sonnecone DM1) and microcrystalline wax was used instead of white soft paraffin and, as regards composition N, triethanolamine was added to the aqueous phase instead of disodium hydrogen phosphate.
• a mixture of Petrolatum Jelly White Sonnecone DM1
• microcrystalline wax was used instead of white soft paraffin
• triethanolamine was added to the aqueous phase instead of disodium hydrogen phosphate.
• Phosphate buffer pH 7.4 Labrasol: ethanol (65: 15:20) 1059 Labrasol in combination with isopropanol
• Phosphate buffer pH 7.4 polysorbate 80: ethanol (65: 15:20) 740
• a skin diffusion experiment was conducted. Full thickness skin from pig ears was used in the study. The ears were kept frozen at - 18°C before use. On the day prior to the experiment the ears were placed in a refrigerator (5 ⁇ 3°C) for slow defrosting. On the day of the experiment, the hairs were removed using a veterinary hair trimmer. The skin was cleaned for subcutaneous fat using a scalpel and two pieces of skin were cut from each ear and mounted on Franz diffusion cells in a balanced order.
• Static Franz-type diffusion cells with an available diffusion area of 3.14 cm 2 and receptor volumes ranging from 8.6 to 1 1.1 ml were used in substantially the manner described by T.J. Franz, "The finite dose technique as a valid in vitro model for the study of percutaneous absorption in man", in Current Problems in Dermatology, 1978, J.W.H. Mall (Ed.), arger, Basel, pp. 58-68. The specific volume was measured and registered for each cell. A magnetic bar was placed in the receptor compartment of each cell. After mounting the skin, physiological saline (35°C) was filled into each receptor chamber for hydration of the skin. The cells were placed in a thermally controlled water bath which was placed on a magnetic stirrer set at 400 rpm.
• the circulating water in the water baths was kept at 35 ⁇ 1°C resulting in a temperature of about 32°C on the skin surface.
• the saline was replaced by receptor medium, 0.04 M isotonic phosphate buffer, pH 7.4 (35°C), containing 4% bovine serum albumin.
• Sink conditions were maintained at all times during the period of the study, i.e. the concentration of the active compounds in the receptor medium was below 10% of the solubility of the compounds in the medium.
• the stratum corneum was collected by tape stripping 10 times using D-Squame tape (diameter 22 mm, CuDerm Corp., Dallas, Texas, USA). Each tape strip is applied to the test area using a standard pressure for 5 seconds and removed from the test area in one gentle, continuous move. For each repeated strop, the direction of tearing off was varied. The viable epidermis and dermis was then sampled from the skin in a similar fashion.
• cathelicidin is an antimicrobial peptide expressed in human keratinocytes.
• the expression of cathelicidin is strongly induced on infection of the skin or disruption of the skin barrier.
• the level of cathelicidin is increased in lesional skin of psoriasis patients. It has been found that the expression of the gene encoding cathelicidin may be induced by vitamin D 3 or vitamin D analogues such as calcipotriol (cf. TT Wang et al, J. Immunol. 173(5), 2004, pp. 2909-2912; J Schauber et al., Immunology 775(4), 2006, pp.
• compositions A, B, C, F, G, I, J, K, L prepared as described in Example 1 above were applied topically in triplicate on reconstructed human epidermis consisting of normal human keratinocytes cultured for 12 days on 0.5 cm 2 polycarbonate filters (available from SkinEthic ® Laboratories, Nice, France) in an amount of 10 ⁇ .
• the tissue was treated for two days followed by separation of the epidermis from the polycarbonate filter and snap-frozen in liquid nitrogen.
• RNA was extracted from the cells and cDNA synthesized by conventional procedures.
• Quantitative real-time PCR was then performed using the following assays from Applied Biosystems: CAMP Hs0018038_ml and GAPDH Hs99999905_ml .
• the expression levels of cathelicidin were normalized to GAPDH and a relative
• composition A 1.9
• composition B 9.4/6.4
• composition K 6.6 composition L 1.9
• compositions of the invention result in higher activation of the target gene, i.e. they may have a higher biological activity in vivo than the marketed ointment.
• compositions B, G, M and N of Example 1 were assessed when administered daily by dermal application to minipigs for 4 weeks. Daivonex ® ointment was used for comparison. Each day the animals were exposed to the test items for 8 hours.
• Clinical signs were recorded daily and skin reactions at the application sites were scored once daily prior to start of dosing and, furthermore, on the day of necropsy in relation to erythema and oedema. Food consumption was recorded daily and the body weight weekly. At the end of the treatment period a gross necropsy was performed on all animals and skin samples were collected from histopathological examination.
• compositions of the invention may be better tolerated in human patients than Daivonex ® ointment.

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