RU2500387C1 - Pharmaceutical composition containing vitamin d analogue and mixed co-solvent and surfactant - Google Patents

Abstract

FIELD: medicine, pharmaceutics.

SUBSTANCE: invention refers to pharmaceutical industry, and represents a local skin composition which is an oil-in-water-in-oil emulsion containing a water phase with a dispersed lipophilic phase containing calcipotriol or monohydrate in the dissolved form; a non-ionic surfactant specified in a group consisting of polyethylene glycol glycerides and C_6-20 fatty acids, polyoxyethylene C_8-20 alkyl ethers or polysorbates and a lower alkanol as a co-solvent; the above water phase is dispersed in a pharmaceutically acceptable anhydrous lipophilic carrier or base.

EFFECT: invention provides the effective dissolution of calcipotriol with the lower content of a lower alcohol, as well as the good penetration of calcipotriol into the viable skin layers, higher biological activity and less skin irritation.

26 cl, 5 ex, 7 tbl, 6 dwg

Description

FIELD OF THE INVENTION

The invention relates to a topical pharmaceutical composition for application to the skin containing a pharmaceutically active agent, a surfactant, a co-solvent and an aqueous phase.

BACKGROUND OF THE INVENTION

Psoriasis is a chronic inflammatory skin disease that manifests itself in the form of erythematous, dry, flaky plaques resulting from hyperkeratosis. Plaques are most often found on the elbows, knees and scalp, although more common lesions may appear on other parts of the body, in particular the lumbosacral region. The most common treatment for mild to moderate psoriasis involves topical application of a composition containing a corticosteroid as an active ingredient. Corticosteroids, although effective, have a drawback due to a number of side effects they cause, such as skin atrophy, acne-like streaks, near-mouth dermatitis, excessive growth of cutaneous fungi and bacteria, pigmented skin hypopigmentation, and rosacea.

However, for many years, the preferred non-steroidal treatment of psoriasis consisted of topical treatment with a vitamin D analog compound, calcipotriol, included in the ointment composition (sold as Daivonex ^® or Dovonex ^® ointment by LEO Pharma), in which calcipotriol is present in the solution or cream composition (sold as Daivonex ^® or Dovonex Cream by LEO Pharma). The solvent in the ointment composition is propylene glycol, which has the advantage of enhancing the penetration of the active ingredient into the skin, resulting in increased effectiveness, but which is also known to act as an irritant to the skin. Thus, it was reported that the inclusion of propylene glycol in local compositions often caused contact dermatitis in patients (in one study, a number of reactions to propylene glycol in the form of irritation, the frequency of which reached 12.5%, see M. Hannuksela et al., Contact Dermatitis 1, 1975, pp. 112-116), and the number of reactions in the form of irritation increases when propylene glycol is used in high concentrations (see review by J. Catanzaro and J. Graham Smith, J. Am. Acad. Dermatol. 24, 1991, pp.90-95). Due to the improved penetration of calcipotriol into the skin, among other reasons due to the presence of propylene glycol, it was found that Daivonex® ointment, more effective in treating psoriatic lesions than Daivonex ^® cream, but also caused skin irritation in a significant part of patients with psoriasis.

Therefore, the object of the invention is to provide a topical composition containing a derivative or analogue of vitamin D as an active ingredient that has skin penetration and biological activity comparable to Daivonex ^® ointment, but which does not contain propylene glycol as a solvent.

SUMMARY OF THE INVENTION

Human skin, in particular the outer layer, the stratum corneum, provides an effective barrier against the penetration of microbial pathogens and toxic chemicals. Although this property of the skin is generally favorable, it complicates the administration of pharmaceuticals through the dermis in that a large amount, if not most of the active ingredient applied to the skin of a patient suffering from a skin disease, may not penetrate into the viable layers of the skin where it is active . To ensure adequate penetration of the active ingredient into the dermis and epidermis as a whole, it is preferable to include the active ingredient in a dissolved state, usually in the presence of a solvent in the form of an alcohol, for example ethanol, or a diol, for example propylene glycol. Propylene glycol is a well-known penetration enhancer, i.e. a substance that is able to penetrate through the stratum corneum and “entrain” low molecular weight components, such as therapeutically active components, in the carrier into the epidermis. Propylene glycol alone can cause significant skin irritation, and it is also able to “carry along” the low molecular weight and potentially irritating components of the carrier into the epidermis, which leads to the general irritating effect of common carriers, including propylene glycol. For this reason, the presence of propylene glycol as a solvent in compositions intended for the treatment of inflammatory skin diseases can exacerbate the inflammatory response.

The purpose of the studies that led to the present invention was to identify a combination of solvents that is more effective for dissolving a poorly soluble compound, such as a vitamin D analog, than low molecular weight alcohols or diols when they are isolated as co-solvents in a mixture with the aqueous phase, and which, additionally contains significantly less co-solvent in the form of low molecular weight alcohol. It has been unexpectedly discovered that mixing certain surfactants with certain lower alkanols provides mixtures with an extremely high solubilization ability. The resulting composition, where the individual solvent components act synergistically, leads to satisfactory penetration of the vitamin D derivative or its analogue into viable layers of the skin at a lower concentration of the co-solvent than when alcohol or diol is used separately as a co-solvent. Additionally, the compositions of the invention exhibit comparable or higher biological activity with respect to Daivonex ^® ointment, as determined by the activation of the target gene described in Example 4 below. Additionally, the composition is physically stable and has a vitamin D analogue that is chemically stable.

Accordingly, the present invention relates to a topical composition for application to the skin, which is an oil-in-water-in-oil emulsion comprising an aqueous phase comprising a lipophilic phase dispersed therein, including

(a) a derivative or analogue of vitamin D in dissolved form;

(b) a nonionic surfactant selected from the group consisting of polyoxyglycerides, derivatives of polyoxyethylene castor oil, polyoxyethylene alkyl ethers, polysorbates or poloxamers; and

(c) lower alkanol as a co-solvent,

wherein said aqueous phase is dispersed in a pharmaceutically acceptable anhydrous lipophilic carrier or base.

In another aspect, the invention relates to a topical composition, as described in this patent application, for the prevention or treatment of skin diseases or conditions.

BRIEF DESCRIPTION OF THE DRAWINGS

Figures 1-4 are graphs showing the solubility of calcipotriol monohydrate in mixtures of a cosolvent with a surfactant included in the composition of the present invention, compared with the solubility of calcipotriol monohydrate, either in a co-solvent or in a surfactant separately.

5a and 5b are graphs showing skin penetration of a composition of the invention.

6 schematically shows the activation of a gene encoding cathelicidin with vitamin D ₃ in human keratinocytes. The cathelicidin gene activation mechanism is used in biological analysis using reconstructed human epidermis (human keratinocytes cultivated to form epidermal layers characteristic of human skin) onto which calcipotriol-containing compositions of the invention are applied to activate cathelicidin, as described in detail below in example 4.

DETAILED DESCRIPTION OF THE INVENTION

Definitions

In the present context, the term “non-ionic surfactant” is intended to mean a surfactant containing a hydrophilic and hydrophobic moiety, where the hydrophobic moiety does not carry a charge but receives its surface activity from highly polar groups such as polyoxyethylene groups. For the purposes of the present invention, the surfactant is a surfactant in the form of an oil-in-water emulsion with an HLB (hydrophilic-lipophilic balance) value of 9-18.

The term "lower alkanol as a co-solvent" is intended to mean a solvent consisting essentially of straight or branched C _1-6 alkanol, for example methanol, ethanol, propanol, isopropanol or butanol.

The term “vitamin D derivative” is intended to mean a biologically active metabolite of vitamin D ₃ , such as calcitriol, or a precursor of a metabolite such as alfacalcidol.

The term “vitamin D analogue” is intended to mean a synthetic compound containing a vitamin D skeleton with side chain modifications and / or skeleton modifications. The analogue exhibits biological activity at the vitamin D receptor, comparable to the biological activity of naturally occurring vitamin D compounds.

"Calcipotriol" is an analogue of vitamin D of the formula

It was found that calcipotriol exists in two crystalline forms, anhydrate and monohydrate. Calcipotriol monohydrate and its preparation are disclosed in WO 94/15912.

The term "storage stability" is intended to mean that the composition exhibits chemical and physical stability characteristics that make it possible to store the composition upon cooling in a refrigerator or, preferably, at room temperature for a sufficient period of time to make the composition commercially viable for example, at least 12 months, in particular at least 18 months, and preferably at least 2 years.

The term “chemical resistance” or “chemically stable” is intended to mean that during storage of the product, usually 2 years, not more than 10%, preferably not more than 5%, of the vitamin D derivative is destroyed. An approximation of chemical resistance at room temperature is obtained conducting accelerated studies of the stability of the composition, which is exposed to a temperature of 40 ° C. If, after storage for 3 months at 40 ° C, less than about 10% of the substance is destroyed, then this is usually accepted as corresponding to a shelf life of 2 years at room temperature. In particular with respect to calcipotriol, the term “chemical resistance” is intended to mean that calcipotriol does not substantially degrade over time to form 24-epi-calcipotriol or other calcipotriol degradation products in the finished pharmaceutical product.

The term “physical stability” or “physically stable” is intended to mean that the composition maintains a macroscopic and microscopic appearance during the shelf life of the product, for example, that a derivative or analog of vitamin D does not precipitate from the solvent phase, or that there is no visible phase separation solvent and carrier phase.

The term "essentially anhydrous" is intended to mean that the content of free water in the lipophilic carrier or base does not exceed about 2% by weight, preferably does not exceed about 1% by weight. carrier or base.

The term "solubilization ability" is intended to mean the ability of a solvent or mixture of solvents to dissolve a given substance, expressed as the amount required to fully solubilize the substance.

The term "synergistic (synergistic)" means that the solubility of a derivative or analogue of vitamin D is significantly higher, in some cases several times higher, when the combination of a co-solvent and a surfactant is present in the aqueous phase than the sum of the solubility values in any of the co-solvent or surfactants when they are added individually to the aqueous phase.

The term "penetration through the skin" is intended to mean the diffusion of the active ingredient into various layers of the skin, i.e. stratum corneum, epidermis and dermis.

The term “skin permeability” is intended to mean the entry of the active ingredient through the skin into the systemic circulation or, in the case of in vitro studies, such as studies, the results of which are presented below in Example 3, of the receptor fluid from the Franz diffusion cell apparatus.

The term "biological activity" is intended to mean the activity of a derivative or analog of vitamin D when applied to the skin in a composition according to the invention. The biological activity of the compositions is determined in an in vitro assay measuring the activation of the target gene encoding cathelicidin in a model of reconstructed human epidermis, including cultured human keratinocytes, as described in detail in Example 5 below.

Embodiments of the invention

In one embodiment, the composition comprises a vitamin D derivative or analog selected from the group consisting of calcipotriol, calcitriol, tacalcitol, maxacalcitol, paricalcitol, and alfacalcidol. In a currently preferred embodiment, the composition comprises calcipotriol or calcipotriol monohydrate as an analogue of vitamin D.

In the composition of the present invention, the surfactant is generally present in a concentration of from about 0.5% by weight. up to about 5% of the mass., or from about 1% of the mass. up to about 3% of the mass., or from about 1.2% of the mass. up to about 2% of the mass., for example, about 1.5% of the mass. per composition.

In accordance with the invention, the nonionic surfactant is preferably selected from the group consisting of polyethylene glycol 8 caprylic / capric acid glycerides (a derivative of polyethylene glycol of a mixture of caprylic and capric acid mono-, di- and triglycerides with an average amount of 8 moles of ethylene oxide) or glycol glycol glyceride 6 / capric acid (a derivative of polyethylene glycol of a mixture of mono-, di- and triglycerides of caprylic and capric acids with an average amount of 6 mol of ethylene oxide. Non-ionic surface but the active agent is preferably polyethylene glycol 8 glyceride of caprylic / capric acids, for example, manufactured by Gattefossé under the tradename Labrasol or company Condea under the trade name Softigen 767.

The nonionic surfactant may also preferably be a polyethylene glycol glyceride of C _6-20 fatty acids selected from the group consisting of caprylocaproyl PEG glyceride, lauroyl PEG glyceride, linoleoyl PEG glyceride, oleoyl PEG glyceride and stearoyl PEG glyceride, simple C _8-20 a polyoxyethylene alkyl ester selected from the group consisting of PEG monoacetyl ether, PEG monolauryl ether, PEG monooleyl ether and PEG monostearyl ether, 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 derivative of castor oil, such as castor oil polyoxyl or hydrogenated castor oil polyoxyl.

As indicated above, the composition further comprises lower alkanol as a co-solvent, which may preferably be selected from the group consisting of methanol, ethanol, n-propanol, isopropanol, n-butanol or 2-butanol. It was unexpectedly discovered that the amount of lower alkanol required for complete dissolution of the derivative or analogue of vitamin D can be significantly reduced (for example, reduced 2-5 times) in the presence of a surfactant, compared with the amount required when lower alkanol used as a solvent. Lower alkanol as a co-solvent can provide a favorable effect at a concentration of about 0.5-5%, in particular about 1-3% or about 2% of the mass. composition.

In a currently preferred embodiment, the co-solvent is ethanol, and the nonionic surfactant is polyethylene glycol 8 caprylic / capric acid glyceride, polysorbate 80 or PEG monoacetyl ether, or the co-solvent is isopropanol, and the nonionic surfactant is castor oil polyoxyl, polyethylene glycol 8 caprylic / capric acid glyceride or polysorbate 80.

The ointment base may be a hydrocarbon or a mixture of hydrocarbons with a chain length in the range of C ₅ to C ₆₀ . An often used ointment base is petroleum jelly or white soft paraffin, which is composed of hydrocarbons with different chain lengths having peaks at about C _40-44 , or a mixture of petroleum jelly and liquid paraffin (consisting of hydrocarbons with different chain lengths, having peaks of C _{28- 40} ). Although petroleum jelly provides occlusion of the treated skin surface, reducing transdermal water loss and potentiating the therapeutic effect of the active ingredient in the composition, it tends to cause a greasy and / or sticky sensation that persists for a fairly long time after application and is not easily distributed over the skin surface. Therefore, it may be preferable to use paraffins consisting of hydrocarbons with a slightly longer chain, such as paraffins consisting of hydrocarbons with a chain length having peaks at C _14-16 , C _18-22 , C _20-22 , C _20-26 , or mixtures thereof (paraffin composition was determined by gas chromatography). It was found that such paraffins are cosmetically more acceptable in that they are less sticky and / or greasy after application and are more easily distributed on the surface of the skin. Therefore, they are expected to lead to improved patient compliance with the prescribed application schedule. Suitable paraffins of this type, referred to as petroleum jelly, are manufactured by Sonneborn and sold under the trade name Sonnecone, for example, 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.

In order to impart a desired viscosity to the composition of the present invention, it is possible to include a lipophilic viscosity increasing ingredient such as wax. The wax may be a mineral wax composed of a mixture of high molecular weight hydrocarbons, for example, saturated C _35-70 alkanes, such as microcrystalline wax. Alternatively, the wax may be a vegetable or animal wax, for example, esters of C _14-32 fatty acids and C _14-32 fatty alcohols, such as beeswax. The amount of viscosity increasing ingredient may vary in accordance with the viscosity giving ability of the ingredient, but may typically be in the range of about 1-20% by weight. composition. When the viscosity increasing ingredient is microcrystalline wax, it is usually present in an amount in the range of about 5-15% by weight, for example, about 10% by weight. composition.

The composition may further comprise an emollient that can act to mitigate the thickened epidermis of psoriatic plaques. A suitable emollient for inclusion in the composition of the present invention may be silicone wax or volatile silicone oil, since it was further found that the presence of silicone facilitates the penetration of calcipotriol into the skin. It has also been found that compositions containing silicone lead to less skin irritation. Suitable silicone oils for inclusion in the composition of the present invention may be selected from cyclomethicone, dimethicone. The amount of silicone oil included in the composition of the present invention, is usually in the range from about 1 to about 10 wt. -%, for example, about 5% of the mass. composition.

It is believed that the presence of propylene glycol in Daivonex ^® ointment is the main factor causing skin irritation experienced by many patients. However, it has been found that calcipotriol alone can cause mild irritation 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). Therefore, it may be preferable to include in the composition of the present invention a compound with an anti-irritant effect, such as glycerin, butylene glycol, sorbitol, sucrose, saccharin, menthol or nicotinamide. Glycerin has been described as a substance that can protect the skin from irritating substances (J. Bettinger et al., Dermatology 197, 1998, pp. 18-24) and, as was found by the applicants, reduces the release of IL-1α in a dose-dependent manner: thus, it was found that the presence of 15% of the mass. glycerol in calcipotriol ointment leads to a significantly lower level of release of IL-1α than the inclusion of 10% of the mass. glycerol, which, in turn, leads to a significantly lower level of release of IL-1α than the inclusion of 5% of the mass. glycerin.

However, in addition to the anti-irritant effect, it was unexpectedly discovered that glycerol is able to potentiate the biological activity of calcipotriol in that it was found that the expression of cathelicidin (in the analysis described below in example 4) increased with decreasing amounts of glycerol in the composition (i.e. more cathelicidin is expressed when the amount of glycerol is 5% by mass than when the amount of glycerol is 10% or 15%, respectively): this implies that in relation to the incorporation of glycerol bend balance between favorable effect with antirazdrazhayuschim potentiating action and advantageous effect. Applicants have found that the inclusion of approximately 5-10% of the mass. glycerol in the composition of the present invention leads to a significant anti-irritant effect, as well as to a significant increase in the biological activity of calcipotriol.

Calcipotriol is known to be a substance that is extremely sensitive to acidic conditions (pH below about 7.0 in aqueous compositions or acidic reactants in non-aqueous compositions) that contribute to the rapid destruction of calcipotriol. To ensure adequate chemical stability of the substance during the entire storage period of the composition, it may be desirable to include a compound capable of neutralizing acidic impurities that may be present in one or more excipients of the composition and which are unfavorable for the chemical stability of calcipotriol. It may be advantageous to select an acid-neutralizing compound from a buffer, such as a phosphate buffer, which can be included in an amount of about 0.025-0.1% by weight. 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.

To maintain good physical stability of the composition, in particular to avoid separation of the aqueous and lipid phases therein, it may be preferable to include a water-in-oil emulsifier with an HLB value of 3-8. Examples of such emulsifiers are polyoxyethylene C _8-22 alkyl ethers, for example polyoxyethylene stearyl ether, polyoxyethylene cetyl ether or polyoxyethylene lauryl ether.

The amount of water in the composition may range from about 1% to about 15% by weight, for example, from about 5% to about 10% by weight. composition.

In a specific embodiment, the composition of the present invention comprises

0.003-0.008% w / w calcipotriol (as monohydrate) 1-3% w / w polyethylene glycol 8 caprylic / capric glyceride 1-3% w / w ethanol 3-8% wt./mass. polyoxyethylene stearyl ether 5-10% wt./mass. water 80-93% wt./mass. paraffin carrier

In another embodiment, the composition of the present invention comprises

0.003-0.008% w / w calcipotriol (as monohydrate) 1-3% w / w polysorbate 80 1-3% w / w ethanol 3-8% wt./mass. polyoxyethylene stearyl ether 5-10% wt./mass. water 80-93% wt./mass. paraffin carrier

In yet another embodiment, the composition of the present invention comprises

0.003-0.008% w / w calcipotriol (as monohydrate) 1-3% w / w polyethylene glycol monoacetyl ether 1-3% w / w ethanol 3-8% wt./mass. polyoxyethylene stearyl ether 5-10% wt./mass. water 80-93% wt./mass. paraffin carrier

In yet another embodiment, the composition of the present invention comprises

0.003-0.008% w / w calcipotriol (as monohydrate) 1-3% w / w polyethylene glycol 8 caprylic / capric glyceride 1-3% w / w isopropanol 3-8% wt./mass. stearyl ether polyoxyethylene 5-10% wt./mass. water 80-93% wt./mass. paraffin carrier

In yet another embodiment, the composition of the present invention comprises

0.003-0.008% w / w calcipotriol (as monohydrate) 1-3% w / w castor oil polyoxyl 1-3% w / w isopropanol 3-8% wt./mass. polyoxyethylene stearyl ether 5-10% wt./mass. water 80-93% wt./mass. paraffin carrier

In yet another embodiment, the composition of the present invention comprises

0.003-0.008% w / w calcipotriol (as monohydrate) 1-3% w / w polysorbate 80 1-3% w / w isopropanol 3-8% wt./mass. polyoxyethylene stearyl ether 5-10% wt./mass. water 80-93% wt./mass. paraffin carrier

The composition of the present invention may also contain other components commonly used in dermal formulations, for example, antioxidants (e.g. alpha-tocopherol), preservatives, sodium edetate, pigments, skin softening agents, skin healing agents and conditioning skin care products skin, such as urea, allantoin, or bisabolol, see CTFA Cosmetic Ingredients Handbook, 2 ^nd Ed., 1992.

The composition of the present invention can also be used in the treatment of psoriasis, sebesporiasis, palmar-plantar pustulosis, dermatitis, ichthyosis, rosacea and acne and related skin diseases by topical application of an effective amount of the composition in accordance with the invention in 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. To this end, the composition in accordance with 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 a derivative or analogue of vitamin D. that a beneficial use of the composition of the present invention is possible for the supportive treatment of said skin diseases, i.e. ongoing treatment after the disappearance of visible symptoms to delay the recurrence of symptoms.

To provide a more effective treatment for psoriasis and other skin conditions in the acute phase, it may be desirable to include one or more additional therapeutically active ingredients in the composition. Examples of such additional active ingredients include, but are not limited to, anti-inflammatory drugs, such as corticosteroids, such as betamethasone and its esters, for example, valerate or dipropionate ester, clobetasol or its esters, such as propionate, hydrocortisone or its esters, such as acetate; non-steroidal anti-inflammatory drugs such as naproxen, indomethacin, diclofenac, ibuprofen, dexibuprofen, ketoprofen, flurbiprofen, piroxicam, tenoxicam, lornoxicam or nabumetone, phosphodiesterase 4 inhibitors or p38 MAP inhibitors.

The invention is further illustrated by the following examples, which are in no way intended to limit the scope of the claimed invention.

Example 1

Compositions of the invention

The compositions shown in tables 1a and 1b below were prepared initially by mixing a surfactant (Cremophor EL, Labrazole, Polysorbate 80 or cetomacrogol 1000) with a co-solvent (ethanol or isopropanol), dissolving calcipotriol monohydrate in the mixture, and finally adding the mixture to an aqueous buffer solution with a pH adjusted to 8.0, and glycerin (when turned on). Then, the resulting dispersion was mixed with a mixture of paraffin, emulsifier (polyoxyethylene stearyl ether), DL-α-tocopherol and sodium edetate.

Table la Compositions A-F in accordance with the invention Composition, mg / g A B C G E F Calcipotriol monohydrate 0,050 0,050 0,050 0,050 0,050 0,050 Disodium hydrogen phosphate 0.65 0.65 0.65 0.65 0.26 0.26 Liquid paraffin fifty fifty fifty fifty fifty fifty Polyoxyethylene Stearyl Ether fifty fifty fifty fifty fifty fifty Purified water 65 65 65 65 65 65 Cetomacrogol 1000 fifteen fifteen Cremophor EL fifteen fifteen Labrazole fifteen fifteen Ethanol twenty twenty twenty twenty Isopropanol twenty twenty Glycerin 85% 65 65 65 Sodium edetate 0,065 0,065 0,065 0,065 0,065 0,065 DL-α-tocopherol 0.02 0.02 0.02 0.02 0.02 0.02 Paraffin white soft up to 1 g up to 1 g up to 1 g up to 1 g up to 1 g up to 1 g

Table lb G-L Compositions of the Invention Composition, mg / g G H I J K L Calcipotriol monohydrate 0,050 0,050 0,050 0,050 0,050 0,050 Disodium hydrogen phosphate 0.65 0.65 0.65 0.65 0.65 0.65 Liquid paraffin fifty fifty fifty fifty fifty fifty Polyoxyethylene Stearyl Ether fifty fifty fifty fifty fifty fifty Purified water 65 65 65 65 65 65 Labrazole fifteen fifteen Polysorbate 80 fifteen fifteen fifteen fifteen Ethanol twenty twenty Isopropanol twenty twenty twenty twenty Glycerin 85% 65 65 65 Sodium edetate 0,065 0,065 0,065 0,065 0,065 0,065 DL-α-tocopherol 0.02 0.02 0.02 0.02 0.02 0.02 Paraffin white soft up to 1 g up to 1 g up to 1 g up to 1 g up to 1 g up to 1 g

table 2 Compositions M and N according to the invention Composition, mg / g M N Calcipotriol monohydrate 0,050 0,050 Disodium hydrogen phosphate 0.65 Triethanolamine one Liquid paraffin fifty fifty Polyoxyethylene Stearyl Ether fifty fifty Purified water 65 65 Polysorbate 80 fifteen Labrazole fifteen Ethanol twenty twenty Glycerin 85% 65 Sodium edetate 0,065 DL-α-tocopherol 0.02 0.02 White Jelly Petroleum (Sonnecon DM1) 699.2 634.2 Microcrystalline Wax (Multiwax 180 MH) one hundred one hundred

Compositions M and N were prepared essentially as described above for AL compositions, except that a mixture of white jelly petroleum (Sonnecon DM1) Petrolatum Jelly White (Sonnecon DM1) and microcrystalline wax was used instead of white soft paraffin, and as for composition N , triethanolamine was added to the aqueous phase instead of disodium hydrogen phosphate.

Example 2

Solubility of calcipotriol in solvent / surfactant mixtures in aqueous buffer

Cetomacrogol 1000 in combination with ethanol

The solubility at 25 ° C of calcipotriol monohydrate, defined as calcipotriol, in an aqueous buffer at pH 7.4 containing cetomacrogol 1000 as a surfactant and ethanol as a co-solvent, is shown in table 3 below. From table 3 it appears that there is a small the synergistic effect of the combination of cetomacrogol 1000 and ethanol on the solubility of calcipotriol in aqueous phosphate buffer solution, i.e. the observed solubility of calcipotriol in an aqueous phosphate buffer containing 15% cetomacrogol 1000 and 20% ethanol is 1468 μg / g, which is approximately 1.4 times higher than the sum of the solubility of calcipotriol in an aqueous phosphate buffer containing either 15% cetomacrogol 1000, or 20% ethanol (1049 μg / g + 2.33 μg / g = 1051 μg / g), see figure 1.

Table 3 Solubility of calcipotriol (in μg / g) in aqueous vehicles using cetomacrogol 1000 as a surfactant and ethanol as a co-solvent at 25 ° C Carrier Solubility, mcg / g Phosphate buffer pH 7.4: ethanol (80:20) 2,33 Phosphate buffer pH 7.4: cetomacrogol 1000 (85:15) 1049 Phosphate buffer pH 7.4: cetomacrogol 1000: ethanol (65:15:20) 1468

Labrazole in combination with ethanol

The solubility at 25 ° C of calcipotriol monohydrate, defined as calcipotriol, in an aqueous buffer at pH 7.4 containing Labrazole as a surfactant and ethanol as a co-solvent, is shown in Table 4 below. From Table 4, it appears that there is a pronounced synergistic the effect of the combination of Labrazole and ethanol on the solubility of calcipotriol in aqueous phosphate buffer solution, i.e. the observed solubility of calcipotriol in an aqueous phosphate buffer containing 15% Labrazole and 20% ethanol is 1059 μg / g, which is approximately 3.6 times higher than the sum of the solubility of calcipotriol in an aqueous phosphate buffer containing either 15% Labrazole or 20% ethanol (292 μg / g + 2.33 μg / g = 294 μg / g), see figure 2.

Table 4 Solubility of calcipotriol (in μg / g) in aqueous vehicles using Labrazole as a surfactant and ethanol as a co-solvent at 25 ° C Carrier Solubility, mcg / g Phosphate buffer pH 7.4: ethanol (80:20) 2,33 PH 7.4 phosphate buffer: labrazole (85:15) 292 Phosphate buffer pH 7.4: labrazole: ethanol (65:15:20) 1059

Labrazole in combination with isopropanol

The solubility at 25 ° C of calcipotriol monohydrate, defined as calcipotriol, in an aqueous buffer at pH 7.4 containing Labrazole as a surfactant and isopropanol as a co-solvent, is shown in Table 5 below. From Table 5, it appears that there is a pronounced synergistic the effect of the combination of labrazole and isopropanol on the solubility of calcipotriol in aqueous phosphate buffer solution, i.e. the observed solubility of calcipotriol in an aqueous phosphate buffer containing 15% Labrazole and 20% isopropanol is 1508 μg / g, which is approximately 4.9 times higher than the sum of the solubility of calcipotriol in an aqueous phosphate buffer containing either 15% Labrazole or 20% isopropanol (292 μg / g + 17.2 μg / g = 309 μg / g), see figure 3.

Table 5 Solubility of calcipotriol (in μg / g) in aqueous vehicles
using labrazole as a surfactant and isopropanol as a co-solvent at 25 ° C Carrier Solubility, mcg / g Phosphate buffer pH 7.4: isopropanol (80:20) 17,2 PH 7.4 phosphate buffer: labrazole (85:15) 292 Phosphate buffer pH 7.4: labrazole: isopropanol (65:15:20) 1508

Polysorbate 80 in combination with ethanol

The solubility at 25 ° C of calcipotriol monohydrate, defined as calcipotriol, in an aqueous buffer at pH 7.4 containing polysorbate (Tween) 80 as a surfactant and ethanol as a co-solvent, is shown below in table 6. From table 6 it is shown that there is a pronounced synergistic effect of the combination of polysorbate 80 and ethanol on the solubility of calcipotriol in aqueous phosphate buffer solution, i.e. the observed solubility of calcipotriol in an aqueous phosphate buffer containing 15% polysorbate 80 and 20% ethanol is 740 μg / g, which is approximately 2.0 times higher than the sum of the solubility of calcipotriol in an aqueous phosphate buffer containing either 15% polysorbate 80, or 20% ethanol (360 μg / g + 2.33 μg / g = 362 μg / g), see Fig. 4.

Table 6 Solubility of calcipotriol (in μg / g) in aqueous vehicles
using polysorbate 80 as a surfactant and ethanol as a co-solvent at 25 ° C Carrier Solubility, mcg / g Phosphate buffer pH 7.4: ethanol (80:20) 2,33 Phosphate buffer pH 7.4: Polysorbate 80 (85:15) 360 Phosphate buffer pH 7.4: Polysorbate 80: Ethanol (65:15:20) 740

The results presented in tables 3-6 show that a higher solubility of calcipotriol monohydrate can be achieved in an aqueous solution by the synergistic action of a co-solvent in combination with a surfactant instead of using either a single co-solvent or one surfactant. This implies that a lower amount of the combination can be used to achieve the same solubility of calcipotriol monohydrate than when each solvent is used separately.

Example 3

Penetration studies

To study the penetration and passage through the skin of calcipotriol from the compositions of the invention, a diffusion experiment was conducted through the skin. In this study, full-thickness skin from pig's ears was used. Ears were kept frozen at -18 ° C before use. On the day before the experiment, the ears were placed in a refrigerator (5 ± 3 ° C) for slow thawing. On the day of the experiment, hair was removed using a veterinary clipper. The skin was cleaned of subcutaneous fat using a scalpel, and two pieces of skin were cut from each ear and placed on Franz diffusion cells in a balanced manner.

Static Franz diffusion cells with an available diffusion area of 3.14 cm ² and receptor volumes ranging from 8.6 to 11.1 ml were used in essentially the same way as described by TJ Franz in The finite dose technique as a valid in vitro model for the study of percutaneous absorption in man ”in the monograph Current Problems in Dermatology, 1978, JWH Mall (Ed.), Karger, Basel, pp. 58-68. For each cell, the specific volume was measured and recorded. A magnetic bar was placed in the receptor compartment of each cell. After skin installation, each receptor chamber was filled with physiological saline (35 ° C) to hydrate the skin. The cells were placed in a temperature-controlled water bath, which was placed on a magnetic stirrer at a set rotation speed of 400 rpm. The temperature of the circulating water in the water baths was maintained at 35 ± 1 ° C, which led to a temperature of about 32 ° C on the skin surface. After one hour, the brine was replaced with receptor medium, 0.04 M isotonic phosphate buffer, pH 7.4 (35 ° C) containing 4% bovine serum albumin. Immersion conditions were maintained at all points in time during the study period, i.e. the concentration of active compounds in the receptor medium was below 10% of the solubility of the compounds in the medium.

In vitro skin penetration of each test composition was tested in 6 repetitions (i.e. n = 6). Each test composition was applied to the skin membrane at 0 hours at the intended dose of 4 mg / cm ² . A glass spatula was used for application, and the residual amount of the composition was determined so as to give the amount of the composition actually applied to the skin.

The skin penetration experiment was allowed to continue for 21 hours. Then the samples were taken from the following compartments:

A sample of the stratum corneum was taken by stripping the top layer with tape 10 times using D-Squame® tape (diameter 22 mm, CuDerm Corp., Dallas, Texas, USA). Each strip of tape was applied to the test area using standard pressure for 5 seconds and removed from the test area in one smooth continuous movement. For each re-strip, the direction of peeling varied. Samples of viable epidermis and dermis were then taken from the skin in a similar manner.

The concentration of calcipotriol in the samples was determined by liquid chromatography mass spectrometry.

The results are presented below on figa and 5b, which shows the amount of calcipotriol found in viable skin (dermis and epidermis) and receptor fluid, in% of the applied dose. An excellent penetration and percolation profile has been found for the compositions of the invention, in particular those containing Labrazole as a surfactant component.

Example 4

The biological activity of the compositions

As shown below in FIG. 6, cathelicidin is an antimicrobial peptide expressed in human keratinocytes. Strong expression of cathelicidin is caused after skin infection or destruction of the skin barrier. With psoriasis, the cathelicidin level increases in the skin of the lesion sites of patients with psoriasis. It has been found that expression of a gene encoding cathelicidin can be caused by analogues of vitamin D ₃ or vitamin D, such as calcipotriol (see TT Wang et al, J. Immunol. 173 (5), 2004, pp. 2909-2912; J Schauber et al., Immunology 118 (4), 2006, pp. 509-519; Schauber and Gallo, J. Allergy Clin Immunol 122, 2008, pp. 261-266; M. Peric et al., PloS One 4 (7 ), July 22, 2009, e6340), by binding to the vitamin D receptor. These data were used to develop an assay in which the uptake and biological activity of calcipotriol was determined by measuring the level of induction of the gene encoding cathelicidin.

In this analysis, compositions A, B, C, F, G, I, J, K, L, obtained as described above in Example 1, were topically applied in triplicate to reconstructed human epidermis consisting of normal human keratinocytes cultured for 12 days on polycarbonate filters with an area of 0.5 cm ² (manufactured by SkinEthic ^® Laboratories, Nice, France) in an amount of 10 μl. The tissue was treated for two days, followed by separation of the epidermis from the polycarbonate filter and instantly frozen in liquid nitrogen. RNA was extracted from cells and cDNA was synthesized by conventional procedures. Quantitative PCR (polymerase chain reaction) (qPCR) was then performed using the following assays from Applied Biosystems: CAMP Hs0018038_m1 and GAPDH Hs99999905_m1. Cathelicidin expression levels were normalized to GAPDH, and relative quantification was performed by comparison with Daivonex® ointment.

The results are presented below in table 7.

Table 7 Composition Multiplicity of activation ¹ Daivonex ^® Ointment 1,0 Composition A 1.9 Composition B 9.4 / 6.4 Composition C 3,5 Composition F 2.1 Composition G 10.9 Composition I 4.1 Composition J 5.2 Composition K 6.6 Composition L 1.9 ¹ Regarding Daivonex® Ointment.

The results presented in table 7 show that the compositions of the invention lead to higher activation of the target gene, i.e. they may have higher in vivo biological activity than a commercially available ointment.

Example 5

Local tolerance study in minipigs

Topical tolerance of the compositions B, G, M, and N of Example 1 was evaluated by daily application of mini-pigs to the skin for 4 weeks. Daivonex ^® ointment was used for comparison. Each day, animals were exposed to the test products for 8 hours.

The study was performed on 10 female Göttingen SPF mini pigs. Each animal had 6 application sites and received a volume of 250 mg of the test formulation per application site. Clinical signs were recorded daily and skin reactions at the sites of application were scored once a day before application and, additionally, on the day of dissection in relation to erythema and edema. Feed intake was recorded daily and body weight weekly. At the end of the treatment period, a macroscopic pathological examination was performed in all animals and skin samples were taken for histopathological examination.

The results show that no adverse clinical signs associated with the treatment were observed during the study, although skin reactions of grade 1-2 (erythema) were observed. Except compositions G, erythema were less pronounced than erythema observed ointment Daivonex ^®. The results indicate that patients can tolerate the compositions of the invention better than Daivonex ^® ointment.

Claims (26)

1. A topical composition for application to the skin, which is an oil-in-water-in-oil emulsion comprising an aqueous phase comprising a lipophilic phase dispersed therein, including
(a) calcipotriol or calcipotriol monohydrate in dissolved form;
(b) a non-ionic surfactant selected from the group consisting of glycerides of polyethylene glycol and C _6-20 fatty acids, C _8-20 alkyl ethers of polyoxyethylene or polysorbates; and
(c) lower alkanol as a co-solvent;
wherein said aqueous phase is dispersed in a pharmaceutically acceptable anhydrous lipophilic carrier or base. 2. The composition according to claim 1, where the non-ionic surfactant is generally present in a concentration of from about 0.5% of the mass. up to about 5% of the mass. in particular, from about 1% of the mass. up to about 3% of the mass. or from about 1.2% of the mass. up to about 2% of the mass., for example, about 1.5% of the mass. per composition. 3. The composition according to claim 1, where the non-ionic surfactant is a polyethylene glycol glyceride of C _6-20 fatty acids selected from the group consisting of caprylocaproyl PEG glyceride, lauroyl PEG glyceride, linoleoyl PEG glyceride, oleoyl PEG glyceride, oleoyl PEG glyceride and stearoyl PEG glyceride , simple C _8-20 alkyl ether of polyoxyethylene, selected from the group consisting of simple monotsetilovogo ester PEG, PEG simple monolauryl ether, simple PEG oleyl ether and PEG simple monostearyl ester, polysorbate selected from the group consisting yaschey of polysorbate 20, 40, 60 and 80. 4. The composition according to claim 1, where the non-ionic surfactant is a polyethylene glycol 8 caprylic / capric acid glyceride or a polyethylene glycol 6 caprylic / capric acid glyceride. 5. The composition according to claim 1, where the lower alkanol as a co-solvent is selected from the group consisting of ethanol, n-propanol, isopropanol. 6. The composition according to claim 5, where the lower alkanol as a co-solvent is present in a concentration of about 0.5-5% wt., In particular about 1-3% wt. or about 2% of the mass. composition. 7. The composition according to claim 1, where the cosolvent is ethanol and the nonionic surfactant is polyethylene glycol 8 caprylic / capric acid glyceride, polysorbate 80 or PEG monoacetyl ether or, where the cosolvent is isopropanol and the nonionic surfactant the substance is polyethylene glycol 8 caprylic / capric acid glyceride or polysorbate 80. 8. The composition according to claim 1, where the carrier includes at least one paraffin selected from the group consisting of hydrocarbons with a chain length from C ₅ to C ₆₀ having peaks at C _14-16 , C _18-22 , C _20-22 , C _20-26 , C _28-40 and C _40-44 (as determined by gas chromatography) or mixtures thereof. 9. The composition according to claim 1, additionally containing an ingredient that increases the viscosity. 10. The composition according to claim 9, where the ingredient that increases the viscosity, is a wax. 11. The composition according to claim 1, additionally containing silicone wax or volatile silicone oil. 12. The composition of claim 11, wherein the volatile silicone oil is cyclomethicone or dimethicone. 13. The composition according to claim 1, additionally containing a compound with anti-irritant effect. 14. The composition of claim 13, wherein the anti-irritant compound is glycerol, butylene glycol, sorbitol, sucrose, saccharin, menthol or nicotinamide. 15. The composition according to claim 1, additionally containing a compound capable of neutralizing acidic impurities that are unfavorable for the chemical resistance of calcipotriol or calcipotriol monohydrate in the composition. 16. The composition of claim 15, wherein said compound is a tertiary amine, such as triethanolamine, trometamol, monoethanolamine or diethanolamine. 17. The composition according to claim 1, containing about 0.001-0.5 mg / g, preferably about 0.002-0.25 mg / g, in particular 0.005-0.05 mg / g calcipotriol or calcipotriol monohydrate. 18. The composition according to claim 1, containing the following ingredients:
0.003-0.008% w / w calcipotriol (as monohydrate) 1-3% w / w polyethylene glycol 8 glyceride caprylic / capric acid 1-3% w / w ethanol 3-8% w / w stearyl ether polyoxyethylene 5-10% w / w water 80-93% of the mass / mass. paraffin carrier 19. The composition according to claim 1, containing the following ingredients:
0.003-0.008% w / w calcipotriol (as monohydrate) 1-3% w / w polysorbate 80 1-3% w / w ethanol 3-8% w / w stearyl ether polyoxyethylene 5-10% w / w water 80-93% of the mass / mass. paraffin carrier 20. The composition according to claim 1, containing the following ingredients:
0.003-0.008% w / w calcipotriol (as monohydrate) 1-3% by weight / mass. monoacetyl ether polyethylene glycol 1-3% w / w ethanol 3-8% about the mass / mass. stearic ether polyoxyethylene 5-10% w / w water 80-93% of the mass / mass. paraffin carrier 21. The composition according to claim 1, containing the following ingredients:
0.003-0.008% w / w calcipotriol (as monohydrate) 1-3% w / w polyethylene glycol 8 glyceride caprylic / capric acid 1-3% w / w isopropanol 3-8% w / w stearic ether polyoxyethylene 5-10% w / w water 80-93% of the mass / mass. paraffin carrier 22. The composition according to claim 1, containing the following ingredients:
0.003-0.008% w / w calcipotriol (as monohydrate) 1-3% w / w polysorbate 80 1-3% w / w isopropanol 3-8% w / w stearic ether polyoxyethylene 5-10% w / w water 80-93% about the mass / mass. paraffin carrier 23. The composition according to any one of paragraphs.18-22, additionally containing 5-10% wt./mass, glycerol. 24. The composition according to claim 1, additionally containing one or more additional therapeutically active ingredients. 25. The composition according to paragraph 24, where such additional active ingredients are selected from the group consisting of anti-inflammatory drugs, such as corticosteroids, such as betamethasone and its esters, for example valerate or dipropionate ester, clobetasol or its esters, such as propionate, hydrocortisone or its esters, such as acetate; non-steroidal anti-inflammatory drugs such as naproxen, indomethacin, diclofenac, ibuprofen, dexibuprofen, ketoprofen, flurbiprofen, piroxicam, tenoxicam, lornoxicam or nabumetone, phosphodiesterase 4 inhibitors or p38 MAP inhibitors. 26. The composition according to claim 1, intended for the treatment of psoriasis.

Images