JP2013515017A - Pharmaceutical composition comprising a vitamin D analog and a co-solvent-surfactant mixture - Google Patents

Abstract

  A topical pharmaceutical composition that is an oil-in-water emulsion containing a vitamin D derivative or analog dissolved in a mixture of a nonionic surfactant and a lower alkanol. The topical pharmaceutical composition may be used for the treatment of skin conditions such as psoriasis.

Description

  The present invention relates to a topical pharmaceutical composition for dermal application consisting of a pharmacologically active agent, a surfactant, a co-solvent and an aqueous phase.

  Psoriasis is a chronic inflammatory skin disease that manifests as erythema-like dry scales resulting from hyperkeratosis. Scale plaques are frequent on the elbows, knees, and scalp, but more extensive lesions can appear in other parts of the body, especially the lumbosacral region. The most common treatment of mild to moderate psoriasis involves the topical application of a composition containing a corticosteroid as the active ingredient. Although effective, corticosteroids have many side effects such as skin atrophy, skin streak, acne-like rash, perioral dermatitis, skin fungal and bacterial overgrowth, depigmentation of colored skin and rosacea There are disadvantages.

  However, for many years, an advantageous non-steroidal treatment for psoriasis has been the ointment composition containing calcipotriol as a solution (sold as LEO Pharma's Daivonex® or Dobonex® ointment) or calcipotriol. There is a topical treatment with calcipotriol, a vitamin D analog compound formulated as a cream composition (sold as LEO Pharma's Daivonex® or Dobonex® cream). The solvent of the ointment composition is propylene glycol, which has the advantage of increasing the penetration of the active ingredient into the skin and providing an enhanced effect, but is also known to act as a skin irritant. Therefore, in topical formulations, the inclusion of propylene glycol frequently causes contact dermatitis in patients (one study that published many irritating responses to 12.5% propylene glycol, M. Hannuksela et al., Contact Dermatitis. 1, 1975, pp. 112-116), the use of propylene glycol at high concentrations increases the number of stimuli-reactants (J. Catanzaro and J. Graham Smith, J. Am. Acad. Dermatol. 24, 1991, pp. 90-95). Due to the increased penetration of calcipotriol into the skin, especially caused by the presence of propylene glycol, Daivonex® ointment is more effective in treating psoriatic lesions than Daivonex® cream, but also considerably It has been found to cause skin irritation in a small proportion of psoriasis patients.

  Accordingly, the present invention provides a topical formulation comprising as an active ingredient a vitamin D derivative or analog that has skin penetration and biological activity properties comparable to Daivonex® ointment but does not contain propylene glycol as a solvent. The purpose is to do.

SUMMARY OF THE INVENTION Human skin, particularly the stratum corneum, the outer layer, provides an effective barrier against the penetration of microbial pathogens and toxic chemicals. While this property of the skin is generally beneficial, a significant, if not most, active ingredient applied on the skin of patients with skin diseases can penetrate the active skin viable layer. This complicates the transdermal administration of pharmaceuticals. In order to ensure sufficient penetration of the active ingredient into the dermis and epidermis, if there is a solvent in the form of an alcohol, it usually dissolves the active ingredient, eg in the presence of ethanol or a diol, eg propylene glycol. It is generally preferred to be included in the state. Propylene glycol is a well-known penetration enhancer, ie, a substance that can penetrate the stratum corneum and “draw” the low molecular weight components that are therapeutically active ingredients in the medium into the epidermis. Propylene glycol can itself cause significant skin irritation, and can “pull” into the epidermis components that are low molecular weight and potentially irritant in the medium, including propylene glycol. Induces the overall stimulating effect of the medium. For this reason, propylene glycol contained as a solvent in the composition to be treated for inflammatory skin diseases may worsen the inflammatory reaction.

In the study leading to the present invention, a combination of solvents that are effective in dissolving poorly soluble compounds such as vitamin D analogues from low molecular weight alcohols or diols as a mixture with an aqueous phase, and as a cosolvent The goal was to express a combination containing a significantly lower amount of low molecular alcohol. It has surprisingly been found that the mixing of a specific surfactant with a specific lower alkanol results in a mixture with an unexpectedly high solubilizing capacity. A composition obtained when the individual solvent components act synergistically than when only alcohol or diol is used as a co-solvent, and satisfies the vitamin D derivative or analog thereof in the skin survival layer with a low concentration of co-solvent. Will lead to a deep penetration. Furthermore, the compositions of the present invention exhibit a biological activity that is comparable to or higher than that of Daivonex® ointment as measured by the target gene activation assay described in Example 4 below. In addition, the composition is physically stable and the vitamin D analog is chemically stable therein.

The present invention therefore relates to a topical composition for dermal application, which comprises
(a) a dissolved form of a vitamin D derivative or analogue;
(b) a nonionic active agent selected from the group consisting of polyoxyl glycerides, polyoxyethylene castor oil derivatives, polyoxyethylene alkyl ethers, polysorbates or poloxamers; and
(c) comprising an aqueous phase in which a lipophilic layer containing a lower alkanol cosolvent is dispersed;
The aqueous phase is dispersed in a pharmaceutically acceptable anhydrous lipophilic carrier or vehicle;
An oil-in-water emulsion.

  In other embodiments, the present invention relates to the topical compositions described herein for use in the prevention or treatment of skin diseases or conditions.

FIG. 2 shows the solubility of calcipotriol monohydrate in the cosolvent-surfactant mixture included in the composition of the present invention, and calcipotriol monohydrate when the cosolvent or surfactant is used alone. It is a graph which compares the solubility of a Japanese thing. Solubility of calcipotriol monohydrate in the cosolvent-surfactant mixture included in the composition of the present invention and solubility of calcipotriol monohydrate when the cosolvent or surfactant is used alone It is a graph which compares. FIG. 3 shows the solubility of calcipotriol monohydrate in the cosolvent-surfactant mixture included in the composition of the present invention, and calcipotriol monohydrate when the cosolvent or surfactant is used alone. It is a graph which compares the solubility of a Japanese thing. FIG. 4 shows the solubility of calcipotriol monohydrate in the co-solvent-surfactant mixture included in the composition of the present invention and the calcipotriol ratio when the co-solvent or surfactant is used alone. It is a graph which compares the solubility of a hydrate. Figures 5a and 5b are graphs showing the penetration of the composition of the present invention into the skin. FIG. 6 is a schematic diagram of activation of a gene encoding cathelicidin by vitamin D3 in human keratinocytes. The mechanism of cathelicidin gene activation is the reconstructed human epidermis (to human skin) to which the calcipotriol-containing composition of the present invention is applied to activate cathelicidin as described in detail in Example 4 below. Used in biological assays using human keratinocytes cultured to form a distinct epidermal layer.

Detailed Description of the Invention In this context, the term "nonionic surfactant" is intended to mean a surfactant comprising hydrophilic and hydrophobic moieties, where the hydrophilic moiety does not carry a charge. However, its surface activity is derived from highly polar groups such as polyoxyethylene groups. For this purpose, the surfactant is an oil-in-water surfactant having an HLB value of 9-18.

The term “lower alkanol co-solvent” is intended to mean a solvent consisting essentially of a C _1-6 linear or branched alkanol, such as methanol, ethanol, propanol, isopropanol or butanol.

  The term “vitamin D derivative” means a biologically active metabolite of vitamin D3 such as calcitriol or a precursor of such a metabolite such as alphacalcidol.

  The term “vitamin D analog” is intended to indicate a synthetic compound comprising a vitamin D backbone with side chain modifications and / or modifications of the backbone itself. Analogs exhibit biological activity comparable to naturally occurring vitamin D compounds on the vitamin D receptor.

のビタミンＤ類似体である。 "Calcipotriol" is a formula

Of vitamin D.

  Calcipotriol has been found to exist in two crystalline forms, an anhydride and a monohydrate. Calcipotriol monohydrate and its preparation are disclosed in WO 94/15912.

  The term “storage stability” is preferably refrigerated for a period of time sufficient for the composition to be commercially valuable, such as at least 12 months, especially at least 18 months and preferably at least 2 years. Is intended to indicate that it has chemical and physical stability properties that allow storage of the composition at room temperature.

  The term “chemical stability” or “chemically stable” means only a vitamin D derivative or analogue that does not exceed 10%, preferably not more than 5%, typically a shelf life of 2 years. It is intended to show that it will not decompose inside. An approximation of chemical stability at room temperature is obtained by subjecting the nanocrystals or compositions containing them to an accelerated stability test at 40 ° C. When degradation after 3 months at 40 ° C. is less than about 10% of the test article, this is usually said to correspond to a shelf life of 2 years at room temperature. In particular, with respect to calcipotriol, “chemical stability” is intended to mean that calcipotriol does not significantly degrade over time into 24-epicalcipotriol or other degradation products in the final drug product. .

  The term “physical stability” or “physically stable” means that the composition retains its macroscopic and microscopic appearance over the shelf life of the product, eg vitamin D derivatives or analogs Means that it does not precipitate from the solvent phase or that there is no two-phase separation of the solvent phase and the carrier phase visible to the naked eye.

  The term “substantially anhydrous” means that the content of free water in the lipophilic carrier or medium does not exceed about 2% by weight, preferably about 1% by weight in the carrier or medium. Intended.

  The term “solubilizing ability” means the ability of a solvent mixture to dissolve a substance (shown as the amount necessary to effect complete solubilization of the substance).

  The term “synergistic” means that when a combination of co-solvent and surfactant is present in the aqueous phase, the vitamin D derivative or the sum of the solubility when the co-solvent or surfactant is individually added to the aqueous phase. It is intended to mean that the solubility of the analog is significantly higher, in some cases several times higher.

  The term “skin penetration” means the diffusion of the active ingredient into various skin layers, namely the stratum corneum, epidermis, dermis.

  The term “skin permeation” means the inflow of the active ingredient into the systemic circulation through the skin or, in the case of the in vitro experiments described in Example 3 below, into the receptor fluid of the Franz cell device used in the experiment.

  The term “biological activity” refers to the activity of a vitamin D derivative or analog when applied to the skin in the composition of the invention. The biological activity of the composition is the activation of a target gene expressing cathelicidin, a biomarker, in a reconstructed human epidermis model for cultured human keratinocytes, as described in detail in Example 5 below. Determined in an in vitro assay.

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

  In the present compositions, the surfactant is generally from about 0.5 to about 5%, or from about 1 to 3% or from about 1.2 to about 2%, such as about 1.5% by weight of the composition. % Concentration.

  According to the present invention, the nonionic active agent is polyethylene glycol 8 caprylic acid / capric glyceride (a polyethylene glycol derivative of caprylic acid and capric acid with an average of 8 moles of mono-, di- and tri-glycerides of ethylene oxide) or Polyethylene glycol 6 caprylic acid / capric glyceride (a polyethylene glycol derivative of caprylic acid and capric acid with a mixture of mono-, di- and tri-glycerides of an average of 6 moles of ethylene oxide). The non-ionic active agent is preferably a polyethylene glycol 8 caprylic / capric glyceride, for example available under the trade name Labrasol from Gattefosse or available under the trade name Softigen 767 from Condea.

The nonionic active agent is preferably a polyethylene glycol C _6-20 fatty acid glyceride selected from the group consisting of caprylocaproyl PEG glyceride, lauroyl PEG glyceride, linoleoyl PEG glyceride, oleoyl PEG glyceride and stearoyl PEG glyceride, PEG mono A polysorbate selected from the group consisting of polyoxyethylene C _8-20 alkyl ether, polysorbate ₂₀ , 40, 60 and 80 selected from the group consisting of cetyl ether, PEG monolauryl ether, PEG monooleyl ether and PEG monostearyl ether A poloxamer selected from the group consisting of poloxamers 124, 237, 338 and 407, or a polyoxyethylene castor oil derivative, such as polyoxyl It may be a better oil or hydrogenated polyoxyl castor oil.

  As noted above, the composition further preferably comprises a lower alkanol co-solvent that can be selected from the group consisting of methanol, ethanol, n-propanol, isopropanol, n-butanol, or 2-butanol. Compared to the amount required when lower alkanol alone is used as the solvent, the amount of lower alkanol required to completely dissolve the vitamin D derivative or analog is substantially reduced in the presence of the surfactant. It has been surprisingly found that it can be obtained (for example 2-5 times reduction). The lower alkanol co-solvent may preferably be present at a concentration of about 0.5 to 5%, especially about 1 to 3% or about 2% by weight of the composition.

  In presently preferred embodiments, the co-solvent is ethanol and the non-ionic active agent is polyethylene glycol 8 caprylic / capric glycerides, polysorbate 80 or PEG monocetyl ether, or the co-solvent is isopropanol and non-ionic The active agent is polyoxyl castor oil, polyethylene glycol 8 caprylic / capric glyceride or polysorbate 80.

Ointments carriers may be a mixture of hydrocarbons or hydrocarbons having a chain length ranging from C ₅ to C _60. Frequently used ointment additives are petrolatum or white soft paraffin or petrolatum and liquid paraffin (various chain lengths of C _28-40 peaks) consisting of hydrocarbons of various chain lengths of about C _40-44 peaks. A mixture of hydrocarbons). Petrolatum occludes the treated skin surface, reduces the transdermal loss of water and enhances the therapeutic effect of the active ingredient in the composition, but remains greasy and / or sticky for a long time after application It tends to have a touch and is not easy to extend. Thus, paraffins consisting of somewhat shorter chain length hydrocarbons, eg, paraffins consisting of hydrocarbons or mixtures thereof where C _14-16 , C _18-22 , C _20-22 , C _20-26 have peak chain lengths It may be preferred to use the class (the hydrocarbon composition of paraffins is determined by gas chromatography). It has been found that such paraffins are cosmetically acceptable because they are less sticky and / or greasy when applied and can be more easily spread. They are therefore expected to improve patient compliance. A suitable paraffin of this type, named Petrolatum Jelly, is manufactured by Sonneborn and marketed under the trade names 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 the desired viscosity to the composition, a lipophilic thickening component such as wax may suitably be included. The wax may be a mineral wax made from a mixture of high molecular weight hydrocarbons such as saturated _C35-70 alkanes such as microcrystalline wax. Alternatively, the wax may be plant or animal waxes such as C14-32 fatty acid esters and _{C 14-32} fatty alcohols, for example, beeswax. The amount of thickening component can vary depending on the viscosifying force of the component, but can typically be in the range of about 1-20% of the composition. When the thickening component is a microcrystalline wax, it is typically present in an amount ranging from about 5 to 15% by weight of the composition, for example about 10%.

  The composition may further comprise an emollient that may act to soften the thickened epidermis of psoriatic plaques. An emollient suitable for addition to the present composition can be silicone wax or volatile silicone oil since it has been further found that the presence of silicone aids the penetration of calcipotriol into the skin. It has also been found that compositions containing silicon result in less cortical irritation. Silicon oils suitable for addition to the composition can be selected from cyclomethicone, dimethicone. The amount of silicone oil added to the composition is typically in the range of about 1 to about 10% by weight of the composition, for example about 5% by weight.

  In Daivonex® ointment, the presence of propylene glycol is believed to be a major causative agent of skin irritation experienced by many patients. However, it has been found that calcipotriol itself can be slightly irritating 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). Thus, it may be advantageous to add anti-irritating compounds such as glycerin, butylene glycol, sorbitol, sucrose, saccharin, menthol or nicotinamide to the composition. 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 we have determined the release of IL-1α in a dose-dependent manner. In this way, the presence of 15% by weight glycerin in calcipotriol ointment significantly reduced the level of IL-1α release from that containing 10% by weight glycerin, and It has also been found to significantly reduce IL-1α release levels over that containing 5 wt% glycerin.

  However, surprisingly, in addition to the anti-irritant effect, reducing the amount of glycerin in the composition enhances cathelicidin expression (the assay described in Example 4 below. Glycerin enhances the biological activity of calcipotriol in that more cathelicidin is expressed when the amount is 5% by weight than when it is 10% or 15% by weight, respectively. It turns out that this can be done: this suggests that a balance must be struck between a favorable anti-irritant effect and a favorable bioactive effect with respect to the addition of glycerin. We have found that the addition of about 5-10% by weight of glycerin to the composition results in a significant anti-irritant effect and a significant increase in the biological activity of calcipotriol.

  Calcipotriol can be very sensitive to the acidic conditions involved in the rapid degradation of calcipotriol (in aqueous compositions, pH below about 7.0 or acidic active substances in non-aqueous compositions). It is a known substance. To assure proper chemical stability of the substance throughout the shelf life of the composition, it can be present in one or more of the composition's additives, neutralizing acidic impurities that are detrimental to the chemical stability of calcipotriol It may be desirable to add a compound that can. The compound that neutralizes the acid may preferably be selected from a buffer such as a phosphate buffer, which may be included in an amount of about 0.025 to 0.1% by weight of the composition. The compound neutralizing the acid may be an amine such as triethanolamine, trometamol, monoethanolamine or diethanolamine. And it can be included in an amount of about 0.1-2% by weight of the composition.

In order to maintain good physical stability of the composition, it may be advantageous to add a water-in-oil emulsifier having an HLB value of 3-8, in particular to avoid separation of the aqueous and lipid phases. Examples of such emulsifiers are polyoxyethylene C _8-22 alkyl ethers such as polyoxyethylene stearyl ether, polyoxyethylene cetyl ether or polyoxyethylene lauryl ether.

  The amount of water in the composition can range from about 1 to about 15%, such as from about 5 to about 10% by weight of the composition.

In certain embodiments, the composition comprises
0.003 to 0.008% w / w calcipotriol (as monohydrate)
1-3% w / w polyethylene glycol 8 caprylic acid / capric glyceride 1-3% w / w ethanol 3-8% w / w polyoxyethylene stearyl ether 5-10% w / w water 80-93% w / w w Includes paraffin carrier.

In other embodiments, the composition comprises:
0.003 to 0.008% w / w calcipotriol (as monohydrate)
1-3% w / w polysorbate 80
1-3% w / w ethanol 3-8% w / w polyoxyethylene stearyl ether 5-10% w / w water 80-93% w / w Contains paraffin carrier.

In yet another aspect, the composition comprises:
0.003 to 0.008% w / w calcipotriol (as monohydrate)
1-3% w / w Polyethylene glycol monocetyl ether 1-3% w / w Ethanol 3-8% w / w Polyoxyethylene stearyl ether 5-10% w / w Water 80-93% w / w Paraffin carrier including.

In still further embodiments, the composition comprises
0.003 to 0.008% w / w calcipotriol (as monohydrate)
1 to 3% w / w polyethylene glycol 8 caprylic acid / capric glyceride 1 to 3% w / w isopropanol 3 to 8% w / w polyoxyethylene stearyl ether 5 to 10% w / w water 80 to 93% w / w w Includes paraffin carrier.

In still further embodiments, the composition comprises
0.003 to 0.008% w / w calcipotriol (as monohydrate)
1-3% w / w Polyoxyl castor oil 1-3% w / w Isopropanol 3-8% w / w Polyoxyethylene stearyl ether 5-10% w / w Water 80-93% w / w Contains paraffin carrier .

In still further embodiments, the composition comprises
0.003 to 0.008% w / w calcipotriol (as monohydrate)
1-3% w / w polysorbate 80
1-3% w / w Isopropanol 3-8% w / w Polyoxyethylene stearyl ether 5-10% w / w Water 80-93% w / w Paraffin carrier.

The composition also includes other ingredients commonly used in skin formulations such as antioxidants (e.g., alpha-tocopherol), preservatives, sodium edetate, pigments, skin soothing agents, skin healing agents and urea, skin conditioning agents may also comprise, such as allantoin or bisabolol ^{(CTFA Cosmetic Ingredients Handbook, 2 nd} Ed., see 1992).

  The composition of the present invention can be administered to a patient in need of treatment by locally administering an effective amount of the composition of the present invention to cause psoriasis, seborrheic psoriasis, palmar plantar pustulosis, dermatitis, ichthyosis, liquor. Can be used to treat acne and acne and related skin diseases. The method preferably comprises topical administration of the composition in a therapeutically sufficient amount once or twice daily. For this purpose, the composition according to the invention preferably contains vitamin D derivatives or analogues from about 0.001 to 0.5 mg / g, preferably from about 0.002 to 0.25 mg / g, in particular 0.005. Contains 0.05 mg / g. It is contemplated that the present compositions may be advantageously used in the maintenance treatment of these skin diseases, ie, continuous treatment to delay the recurrence of symptoms after disappearance of visible symptoms.

  It may be desirable to include one or more additional therapeutically active ingredients in the composition to provide effective treatment of acute psoriasis and other skin conditions. Examples of such further active ingredients are anti-inflammatory agents such as corticosteroids, for example betamethasone and its esters, such as valeric acid or dipropionate, clobetasol or its esters, such as propionate, hydrocortisone or its esters, such as acetic acid. Esters; including non-steroidal anti-inflammatory agents such as naproxen, indomethacin, diclofenac, ibuprofen, dexibprofen, ketoprofen, flurbiprofen, piroxicam, tenoxicam, lornoxicam or nabumetone, phosphodiesterase 4 inhibitors or p38 MAP kinase inhibitors It is not limited to these.

  The invention is further illustrated by the following examples, which are not intended to limit the scope of the invention as claimed in any way.

Example 1
Compositions of the Invention The compositions shown in Tables 1a and 1b below are first mixed with a surfactant (Cremophor EL, Labrasol, polysorbate 80 or cetomacrogol 1000) and a co-solvent (ethanol or isopropanol) Potriol monohydrate was dissolved in the mixture and finally prepared by adding the mixture to aqueous buffer adjusted to pH 8.0 and glycerin (if included). The resulting dispersion was mixed with a mixture of paraffin, emulsifier (polyoxyethylene stearyl ether), DL-α-tocopherol and sodium edetate.

組成物ＭおよびＮは、本質的にペトロラタム・ゼリー白色(Sonnecone DM1)および微結晶蝋の混合物を白色軟パラフィンの代わりに使用し、組成物Ｎに関しては、トリエタノールアミンをリン酸水素二ナトリウムの代わりに水相に添加した以外、組成物Ａ〜Ｌについて上記した通りに製造した。

Compositions M and N use essentially a mixture of petrolatum jelly white (Sonnecone DM1) and microcrystalline wax instead of white soft paraffin, and for composition N triethanolamine is disodium hydrogen phosphate. Instead, it was prepared as described above for compositions A to L except that it was added to the aqueous phase.

Example 2
Solubility of Calcipotriol in Solvent / Surfactant Mixture of Aqueous Buffer Setomacrogol 1000 Combined with Ethanol
Table 3 below shows the solubility of calcipotriol monohydrate measured as calcipotriol in aqueous buffer pH 7.4 containing cetomacrogol 1000 as surfactant and ethanol as cosolvent at 25 ° C. Show. From Table 3, there is a slight synergistic effect of the combination of cetomacrogol 1000 and ethanol on the solubility of calcipotriol in aqueous phosphate buffer, ie, aqueous phosphorus containing 15% cetomacrogol 1000 and 20% ethanol. The observed solubility of calcipotriol in acid buffer is 1468 μg / g, which is the solubility of calcipotriol in aqueous phosphate buffer containing either 15% cetomacrogol 1000 or 20% ethanol. It is shown to be about 1.4 times higher than the sum of (1049 μg / g + 2.33 μg / g = 1051 μg / g) (see FIG. 1).

Labrasol combined with ethanol
Table 4 below shows the solubility of calcipotriol monohydrate measured as calcipotriol in an aqueous buffer pH 7.4 containing Labrasol as a surfactant and ethanol as a cosolvent at 25 ° C. From Table 4, there is a significant synergy with the combination of Labrasol and ethanol for the solubility of calcipotriol in aqueous phosphate buffer, ie aqueous phosphate buffer calcipotri containing 15% Labrasol and 20% ethanol. The observed solubility of all is 1059 μg / g, which is about 3.6 times higher than the sum of the solubility of calcipotriol in aqueous phosphate buffer containing either 15% Labrasol or 20% ethanol. (292 μg / g + 2.33 μg / g = 294 μg / g) (see FIG. 2).

Labrasol combined with isopropanol
Table 5 below shows the solubility of calcipotriol monohydrate measured as calcipotriol in aqueous buffer pH 7.4 containing Labrasol as a surfactant and isopropanol as a co-solvent at 25 ° C. From Table 5, it can be seen that there is significant synergy with the combination of Labrasol and isopropanol for the solubility of calcipotriol in aqueous phosphate buffer, ie, aqueous phosphate buffer calcipotri containing 15% Labrasol and 20% isopropanol. The observed solubility of oar is 1508 μg / g, which is about 4.9 times higher than the sum of the solubility of calcipotriol in aqueous phosphate buffer containing either 15% Labrasol or 20% isopropanol. (292 μg / g + 17.2 μg / g = 309 μg / g) is shown (see FIG. 3).

Polysorbate 80 combined with ethanol
Table 6 below shows the solubility of calcipotriol monohydrate measured as calcipotriol in aqueous buffer pH 7.4 containing polysorbate (Tween 80 as surfactant and ethanol as cosolvent) at 25 ° C. Indicates. Table 6 shows that the solubility of calcipotriol in aqueous phosphate buffer is synergistic with the combination of polysorbate (Tween 80) and ethanol, ie, aqueous phosphorus containing 15% polysorbate (Tween 80) and 20% ethanol. The observed solubility of the acid buffer calcipotriol is 740 μg / g, which is that of the aqueous phosphate buffer calcipotriol containing either 15% polysorbate (Tween 80) or 20% ethanol. It is shown to be about 2.0 times higher than the sum of the solubility (360 μg / g + 2.33 μg / g = 362 μg / g) (see FIG. 4).

  The results shown in Tables 3-6 above show that, instead of using only the co-solvent or only the surfactant, the synergistic effect of the co-solvent in combination with the surfactant results in calcipotriol monohydrate in the aqueous phase. It shows that it is possible to increase the solubility. This means that the same solubility of calcipotriol monohydrate can be achieved with a smaller volume combination than when using either solvent alone.

Example 3
Penetration test To test the skin penetration and permeation of calcipotriol from the composition of the invention, a skin diffusion experiment was performed. A sufficiently thick skin from pig ears was used in this experiment. Ears were frozen at −18 ° C. before use. The day before the experiment, the ears were placed in a refrigerator (5 ± 3 ° C.) for slow thawing. On the day of the experiment, the hair was removed using a veterinary hair trimmer. Subcutaneous fat was removed from the skin using a scalpel and two pieces of skin were excised from each ear and mounted in a Franz diffusion cell in order of equilibrium.
Substantially TJ 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, JWH Mall (Ed.), Karger, Basel, pp. 58- A static Franz type diffusion cell with an available diffusion area of 3.14 cm ² and a receptor volume in the range of 8.6 to 11.1 ml was used as described in 68. A specific volume was measured and registered for each cell. A magnetic bar was provided in the receptor compartment of each cell. After attaching the skin, saline (35 ° C.) was filled into each receptor chamber for skin hydration. The cell was placed in a thermally controlled water bath placed on top of a magnetic stirrer set at 400 rpm. Circulating water in the water bath was maintained at 35 ± 1 ° C. and the skin surface temperature was about 32 ° C. After 1 hour, the saline was changed to receptor medium, 0.04 M isotonic phosphate buffer, 4% bovine serum albumin, pH 7.4 (35 ° C.). Sedimentation conditions were maintained throughout the test, i.e. the concentration of active compound in the receptor fluid was less than 10% of the solubility of the compound in the medium.

The in vitro skin penetration of each test composition was tested in 6 replicates (ie n = 6). Each test composition was applied to the skin membrane at 0 hours at the intended 4 mg / cm ² dose. A glass spatula was used for the application and the remaining amount of the composition was determined to obtain the amount of composition actually applied to the skin.
The skin penetration experiment was allowed to proceed for 21 hours. Samples were then collected from the following compartments:
The stratum corneum was collected by stripping the tape 10 times using D-Squame® tape (22 mm diameter, CuDerm Corp., Dallas, Texas, USA). Each piece of tape was applied to the test area using standard pressure for 5 seconds and removed from the test area with one gentle, continuous movement. The direction of peeling was changed for repeated peeling. Viable epidermis and dermis were then sampled from the skin in a similar manner. A sample (1 ml) of the receptor fluid in the diffusion cell was collected and analyzed.
The calcipotriol concentration of the sample was determined by LC mass spectrometry.
The results are evident from the accompanying FIGS. 5a and 5b, which show the amount of calcipotriol found in viable skin (dermis and epidermis) and receptor fluid as a percentage of the applied dose. The compositions of the present invention, particularly those containing Labrasol as a surfactant component, have been found to have excellent penetration and permeation profiles.

Example 4
Biological activity of the composition As shown in FIG. 6 below, cathelicidin is an antimicrobial peptide expressed in human keratinocytes. Cathelicidin expression is strongly induced by skin infection or skin barrier breakdown. In psoriasis, cathelicidin levels increase in skin lesions in psoriasis patients. Through binding to the vitamin D receptor, it has been found that expression of the gene encoding cathelicidin can be induced by analogues of vitamin D such as vitamin D3 or calcipotriol (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). This finding has been exploited in the development of assays in which calcipotriol uptake and biological activity from test compositions in human keratinocytes is determined by measuring the induction level of the gene encoding cathelicidin.

^１ Daivonex(登録商標)軟膏に対する
表７に示す結果は、本発明の組成物が標的遺伝子のより高い活性をもたらす、すなわち、それらは、市販の軟膏よりインビボで高い生物学的活性を有することを示す。 In this assay, compositions A, B, C, F, G, I, J, K, L, prepared as described in Example 1 above, were added in a volume of 10 μl to a 0.5 cm ² polycarbonate filter ( Three times were applied topically to reconstructed human epidermis consisting of normal human keratinocytes cultured on SkinEthic® Laboratories, Nice, France) for 12 days. The tissue was treated for 2 days, the epidermis was separated from the polycarbonate filter and snap frozen in liquid nitrogen. RNA was extracted from the cells and cDNA was synthesized by conventional procedures. Quantitative real-time PCR (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 shown in Table 7 below.

^{1 for} Daivonex® ointment The results shown in Table 7 show that the compositions of the present invention provide higher activity of the target gene, ie, they have higher biological activity in vivo than the commercial ointment. Show.

Example 5
Mini-Pig Local Tolerance Test The local tolerability of compositions B, G, M and N of Example 1 was assessed when administered transdermally to mini-pigs daily for 4 weeks. Daivonex® ointment was used for comparison. Each day, the animals were exposed to the test substance for 8 hours.
The experiment was performed with 10 female Goettingen SPF minipigs. Six sites were applied to each animal, and an amount of 250 mg test formulation per site was applied. Clinical symptoms were recorded daily, and skin reaction at the site of application was assessed once daily prior to initiation of administration for erythema and edema and further on the day of necropsy. Food consumption was recorded daily and body weight was recorded weekly. At the end of the treatment period, a complete necropsy was performed on all animals and skin samples were collected by histopathology.
The results show that grade 1-2 skin reactions (erythema) were observed, but no adverse treatment-related clinical signs were observed during the experiment. Except for Composition G, erythema was less expressed than observed with Daivonex® ointment. This result means that the composition of the present invention can be tolerated in human patients more than Daivonex® ointment.

In the study leading to the present invention, a combination of solvents that are effective in dissolving poorly soluble compounds such as vitamin D analogues from low molecular weight alcohols or diols as a mixture with an aqueous phase, and as a cosolvent it was an object of out have seen a combination comprising a low molecular alcohol significantly lower amounts. It has surprisingly been found that the mixing of a specific surfactant with a specific lower alkanol results in a mixture with an unexpectedly high solubilizing capacity. Than when using only alcohol or diol as co-solvent, satisfaction of the individual composition from which the solvent is obtained when the act synergistically, low concentrations of vitamin D derivative to the skin survival layer in a co-solvent or an analogue thereof Will lead to a deep penetration. Furthermore, the compositions of the present invention exhibit a biological activity that is comparable to or higher than that of Daivonex® ointment as measured by the target gene activation assay described in Example 4 below. In addition, the composition is physically stable and the vitamin D analog is chemically stable therein.

FIG. 1 shows the solubility of calcipotriol monohydrate in the cosolvent-surfactant mixture contained in the composition of the present invention, and calcipotriol monohydrate when the cosolvent or surfactant is used alone. It is a graph which compares the solubility of a Japanese thing. FIG. 2 shows the solubility of calcipotriol monohydrate in the cosolvent-surfactant mixture included in the composition of the present invention, and calcipotriol monohydrate when the cosolvent or surfactant is used alone. It is a graph which compares the solubility of a Japanese thing. FIG. 3 shows the solubility of calcipotriol monohydrate in the cosolvent-surfactant mixture included in the composition of the present invention, and calcipotriol monohydrate when the cosolvent or surfactant is used alone. It is a graph which compares the solubility of a Japanese thing. FIG. 4 shows the solubility of calcipotriol monohydrate in the co-solvent-surfactant mixture included in the composition of the present invention and the calcipotriol ratio when the co-solvent or surfactant is used alone. It is a graph which compares the solubility of a hydrate. Figures 5a and 5b are graphs showing the penetration of the composition of the present invention into the skin. FIG. 6 is a schematic diagram of activation of a gene encoding cathelicidin by vitamin D3 in human keratinocytes. The mechanism of cathelicidin gene activation is the reconstructed human epidermis (to human skin) to which the calcipotriol-containing composition of the present invention is applied to activate cathelicidin as described in detail in Example 4 below. Used in biological assays using human keratinocytes cultured to form a distinct epidermal layer.

Claims (30)

A topical composition for dermal application comprising:
(a) a dissolved form of a vitamin D derivative or analogue;
(b) a nonionic active agent selected from the group consisting of polyoxyl glycerides, polyoxyethylene castor oil derivatives, polyoxyethylene alkyl ethers, polysorbates or poloxamers; and
(c) comprising an aqueous phase in which a lipophilic layer containing a lower alkanol cosolvent is dispersed;
The aqueous phase is dispersed in a pharmaceutically acceptable anhydrous lipophilic carrier or vehicle;
A composition, which is an oil-in-water emulsion.   The composition according to claim 1, wherein the vitamin D derivative or analogue is selected from the group consisting of calcipotriol, calcitriol, tacalcitol, maxacalcitol, paricalcitol and alphacalcidol.   The composition according to claim 2, wherein the vitamin D analog is calcipotriol or calcipotriol monohydrate.   The nonionic active agent is present in a total concentration of about 0.5 to about 5% by weight of the composition, in particular about 1 to about 3% or about 1.2 to about 2%, for example about 1.5%. The composition according to claim 1, which is present in any one of claims 1 to 3. Polyethylene glycol C _6-20 fatty acid glyceride, PEG monocetyl ether, wherein the nonionic active agent is selected from the group consisting of caprylocaproyl PEG glyceride, lauroyl PEG glyceride, linoleoyl PEG glyceride, oleoyl PEG glyceride and stearoyl PEG glyceride; Polysorbate selected from the group consisting of PEG monolauryl ether, PEG monooleyl ether and PEG monostearyl ether, polysorbate selected from the group consisting of polysorbate ₂₀ , 40, 60 and 80, poloxamer 124 Poloxamer or polyoxyethylene castor oil derivative selected from the group consisting of 237, 338 and 407, such as polyoxyl castor oil or A curing polyoxyl castor oil, composition according to any one of claims 1 to 4.   The composition according to any one of claims 1 to 5, wherein the nonionic active agent is polyethylene glycol 8 caprylic acid / capric glyceride or polyethylene glycol 6 caprylic acid / capric glyceride.   7. A composition according to any one of claims 1 to 6, wherein the lower alkanol cosolvent is selected from the group consisting of methanol, ethanol, n-propanol, isopropanol, n-butanol or 2-butanol.   8. A composition according to claim 7, wherein the lower alkanol cosolvent is present at a concentration of about 0.5-5%, in particular 1-3% or about 2% by weight of the composition.   The co-solvent is ethanol and the non-ionic active agent is polyethylene glycol 8 caprylic / capric glyceride, polysorbate 80 or PEG monocetyl ether, or the co-solvent is isopropanol and the non-ionic active agent is polyoxyl The composition according to any one of claims 1 to 8, which is castor oil, polyethylene glycol 8 caprylic / capric glyceride or polysorbate 80. Carrier a chain length ranging from _{C 5} to _{C 60, C 14-16, C 18-22} , C 20-22, _{paraffins C 20-26} consists of hydrocarbons or mixtures thereof having a chain length of peak The composition of claim 1 comprising a class (determined by gas chromatography).   Furthermore, the composition of any one of Claims 1-10 containing a thickening component.   12. A composition according to claim 11 wherein the thickening component is a wax.   The composition according to any one of claims 1 to 12, further comprising silicone wax or volatile silicone oil.   14. The composition of claim 13, wherein the volatile silicone oil is cyclomethicone or dimethicone.   The composition according to any one of claims 1 to 14, further comprising an anti-irritant compound.   16. The composition of claim 15, wherein the anti-irritant compound is glycerin, butylene glycol, sorbitol, sucrose, saccharin, menthol or nicotinamide.   17. A composition according to any one of the preceding claims, further comprising a compound capable of neutralizing acidic impurities that are detrimental to the chemical stability of the vitamin D derivative or analog.   18. A composition according to claim 17, wherein the compound is a tertiary amine such as triethanolamine, trometamol, monoethanolamine or diethanolamine.   19. A vitamin D derivative or analogue according to claim 1 comprising about 0.001 to 0.5 mg / g, preferably about 0.002 to 0.25 mg / g, in particular 0.005 to 0.05 mg / g. The composition according to any one of the above. 0.003 to 0.008% w / w calcipotriol (as monohydrate)
1-3% w / w polyethylene glycol 8 caprylic acid / capric glyceride 1-3% w / w ethanol 3-8% w / w polyoxyethylene stearyl ether 5-10% w / w water 80-93% w / w w A composition according to any one of the preceding claims comprising a paraffin carrier. 0.003 to 0.008% w / w calcipotriol (as monohydrate)
1-3% w / w polysorbate 80
20. 1 to 3% w / w ethanol 3 to 8% w / w polyoxyethylene stearyl ether 5 to 10% w / w water 80 to 93% w / w Paraffin carrier. A composition according to 1. 0.003 to 0.008% w / w calcipotriol (as monohydrate)
1-3% w / w Polyethylene glycol monocetyl ether 1-3% w / w Ethanol 3-8% w / w Polyoxyethylene stearyl ether 5-10% w / w Water 80-93% w / w Paraffin carrier The composition of any one of Claims 1-19 containing this. 0.003 to 0.008% w / w calcipotriol (as monohydrate)
1 to 3% w / w polyethylene glycol 8 caprylic acid / capric glyceride 1 to 3% w / w isopropanol 3 to 8% w / w polyoxyethylene stearyl ether 5 to 10% w / w water 80 to 93% w / w w A composition according to any one of the preceding claims comprising a paraffin carrier. 0.003 to 0.008% w / w calcipotriol (as monohydrate)
1-3% w / w Polyoxyl castor oil 1-3% w / w Isopropanol 3-8% w / w Polyoxyethylene stearyl ether 5-10% w / w Water 80-93% w / w Contains paraffin carrier The composition of any one of Claims 1-19. 0.003 to 0.008% w / w calcipotriol (as monohydrate)
1-3% w / w polysorbate 80
1 to 3% w / w Isopropanol 3 to 8% w / w Polyoxyethylene stearyl ether 5 to 10% w / w Water 80 to 93% w / w Paraffin carrier. A composition according to 1.   26. The composition of any one of claims 20-25, further comprising 5-10% w / w glycerin.   27. A composition according to any one of claims 1 to 26, further comprising one or more additional therapeutically active ingredients.   Further active ingredients are anti-inflammatory agents such as corticosteroids, such as betamethasone and its esters, such as valeric acid or dipropionate, clobetasol or its esters, such as propionate, hydrocortisone or its esters, such as acetate; naproxen Selected from non-steroidal anti-inflammatory agents such as indomethacin, diclofenac, ibuprofen, dexuibuprofen, ketoprofen, flurbiprofen, piroxicam, tenoxicam, lornoxicam or nabumetone, phosphodiesterase 4 inhibitors or p38 MAP kinase inhibitors 28. The composition according to 27.   29. A composition according to any one of claims 1 to 28 for use in the treatment of a skin disease or condition.   30. The composition of claim 29, wherein the skin disease or condition is psoriasis, seborrheic psoriasis, palmar plantar pustulosis, dermatitis, ichthyosis, rosacea or acne.

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