Calcipotriol monohydrate nanocrystals

FIELD: medicine, pharmaceutics.

SUBSTANCE: invention refers to pharmaceutics and represents a suspension for treating psoriasis, containing calcipotriol monohydrate in the form of nanocrystals having the particle size distribution within the range of 200-600 nm; the particles are dispersed in an aqueous phase containing a non-ionic polymer surfactant specified in a group consisting of a surfactant in the form of poloxamers or polysorbates, in the amount of 0.01-5 wt % calculated using a suspension for preventing development of aggregation and/or calcipotriol monohydrate nanocrystal growth; the calcipotriol monohydrate nanocrystals are produced in the suspension by processing the suspension by a method involving the stages of reduction in crystalline calcipotriol monohydrate particle size in an aqueous phase to form microparticles having the particle size distribution within the approximate range of 5-20 mcm and the average approximate particle size of 10 mcm; the suspension is exposed to three high-pressure homogenisation cycles for 7-15 minutes each; in the first, second and third cycles, the pressure makes 300-800 bars, 800-1,200 bars and 1,200-1,700 bars respectively.

EFFECT: invention provides creating the local composition containing calcipotriol as an active agent, however being free from propylene glycol as a solvent.

34 cl, 8 ex, 5 tbl, 9 dwg

 

The technical field TO WHICH the INVENTION RELATES

The present invention relates to calcipotriol the monohydrate in the form of nanocrystals and the nanocrystals in a pharmaceutical composition intended for the prevention or treatment of dermal diseases and conditions.

Background of the INVENTION

Psoriasis is a chronic inflammatory skin disease, which manifests itself in the form of erythematous dry scaly plaques resulting from hyperkeratosis. Plaques most often found on the elbows, knees and hairy scalp, although you may see more widespread lesions on other parts of the body, in particular, the lumbar-sacral region. The most common treatment of psoriasis of mild to moderate severity, and includes local application of a composition containing a corticosteroid as the active ingredient. Corticosteroids are effective tools, but have the disadvantage of a number of side effects such as skin atrophy, stripes, prepodobnye rash, perioral dermatitis, overgrowth of skin of bacteria and fungi, hypopigmentation pigmented skin and acne rosacea.

However, for many years the preferred non-steroidal anti-psoriasis treatment consisted in the topical treatment of the connection of an analog of vitamin�on D, calcipotriol included in the composition of the ointment compositions (sold as an ointment Daivonex® or Dovonex® company LEO Pharma), which calcipotriol is present in the solution or cream composition (sold in the form of creams Daivonex® or Dovonex® company LEO Pharma), which calcipotriol is present in the form of suspension or microparticles. The solvent in the wax composition is propylene glycol, which has the advantage of enhancing the penetration of active ingredient into the skin, which leads to greater efficiency, but which also, as is known, acts as a skin irritant. So, it was reported that the inclusion of propylene glycol in local composition often causes the development of contact dermatitis patients (in one study it was reported the frequency of reactions irritation propylene glycol, reaching 12.5 percent, see M. Hannuksela et al., Contact Dermatitis 1, 1975, pp. 112-116), and the number of reactions of irritation increases when propylene glycol is used in high concentrations (see review of J. Catanzaro and J. Graham Smith, J. Am. Acad. Dermatol. 24, 1991, pp. 90-95). Due to improved penetration of calcipotriol in the skin as a result, along with other reasons for the presence of propylene glycol, it was found that the ointment Daivonex® is more effective in treating psoriatic lesions than Daivonex cream®but it also caused skin irritation in a significant proportion of patient� with psoriasis.

A BRIEF 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 generally, it complicates the introduction of pharmaceuticals through the skin to the fact that a large number, if not most, of the active ingredient applied to the skin of a patient suffering from skin disease, may not penetrate into the viable layers of the skin where it exerts its activity. To ensure adequate penetration of active ingredients into the dermis and the epidermis, in General, preferably the inclusion of the active ingredient in a dissolved state, usually in the presence of a solvent in the form of an alcohol, e.g. ethanol, or a diol, such as propylene glycol. Propylene glycol is a well-known amplifier penetration, i.e. a substance that is able to penetrate through the stratum corneum and to inspire a" low molecular weight components, such as therapeutically active components in the media, in the epidermis. The propylene glycol causes significant irritation to the skin and is able to captivate with a" low-molecular annoying media components in the epidermis, causing General annoyance to the effect of conventional media, including�I propylene glycol. For this reason, the presence of propylene glycol as a solvent in the compositions intended for the treatment of inflammatory skin disease, may exacerbate the inflammatory response.

The objective of the study that underlies the present invention consisted in providing a local compositions containing calcipotriol as active ingredient, which has the properties of skin penetration and biological activity, comparable with the properties of the ointment Daivonex®, but which does not contain propylene glycol as a solvent.

Unexpectedly it was discovered that it was possible to obtain calcipotriol monohydrate in the form of nanocrystals, which are chemically stable (i.e., do not decompose in 24-EPI calcipotriol or other products of decomposition), because, unexpectedly, do not form significant quantities of amorphous calcipotriol as a result of high force or impact forces or high temperatures during the bringing down to the nano range. In addition, the nanocrystals are physically stable, with no observed aggregation or crystal growth or change in the shape of crystals (polymorphic) in suspensions of nanocrystals after receipt. The nanocrystals are easily included in compositions for topical application in the form of cream and ointment, calcipotriol (monohydrate) may �to sonicate in viable skin layers (i.e., the dermis and epidermis) in quantities comparable with the penetration of calcipotriol of ointment Daivonex®, and lead to similar or higher levels of biological activity (according to the definition of activation of the target genesin vitro) without turning the amplifier penetration, such as propylenglycol, which is a potential skin irritant.

Accordingly, in one aspect, the present invention relates to suspensions of calcipotriol monohydrate in the form of nanocrystals with particle size distributions in the range 200-600 nm, according to the definition of dynamic light scattering, wherein the suspension further comprises a water phase comprising non-ionic, polymeric surfactant in an amount sufficient to prevent formation of aggregates and/or crystal growth of nanocrystals calcipotriol monohydrate.

In another aspect, the invention relates to calcipotriol monohydrate with the distribution of particle size in the range 200-600 nm, according to the definition of dynamic light scattering, moreover, these nanocrystals can be obtained by a method comprising the stage

(a) reduction of crystalline calcipotriol monohydrate in an aqueous phase containing non-ionic, polymeric surfactant in an amount in the range from about 1% to p�around 5% of the mass. in the calculation of the aqueous phase, with the formation of microparticles with a distribution of particle size in the range of about 5-20 μm and an average particle size of about 10 microns;

(b) effect on the suspension of step (a), the first cycle of homogenization under high pressure, comprising of about 300-800 bar for a period of time sufficient to obtain approximately 15-40% of crystals of calcipotriol monohydrate with the distribution of particle size in the range of 200-600 nm;

(c) effect on the slurry of step (b), the second cycle of homogenization under high pressure, constituting approximately 800-1200 bar, for a time period sufficient to obtain approximately 40-80% of the crystals calcipotriol monohydrate with the distribution of particle size in the range of 200-600 nm;

(d) effect on the slurry of step (c), the third cycle of homogenization under high pressure, constituting approximately 1200-1700 bar, during a period of time sufficient to obtain about 90% or more of crystals of calcipotriol monohydrate with the distribution of particle size in the range of 200-600 nm; and

(e) optionally, separation of the obtained nanocrystals calcipotriol monohydrate from the aqueous phase.

In another aspect the invention relates to pharmaceutical compositions containing the above-described nanocrystals calcipotriol mo�of agitate and a pharmaceutically acceptable carrier.

In another aspect the invention relates to the use of compositions containing nanocrystals or nanosuspension of calcipotriol monohydrate for the treatment of diseases or skin conditions such as psoriasis, sebopsoriasis, Palmar-plantar pustulosis, dermatitis, ichthyosis, or pimples.

BRIEF description of the DRAWINGS

Fig.1 presents a graph showing the particle size distributions of the nanocrystals calcipotriol monohydrate obtained by the method according to the present invention, according to the definition of dynamic light scattering.

Fig.2a presents a graph that compares the Raman spectrum of nanosuspension of calcipotriol monohydrate in 2% poloxamer 188 with the Raman spectrum of calcipotriol monohydrate, not subjected to bring to the nano range. The graph shows that the method of bringing to the nano range in accordance with the invention does not lead to any change in the form of crystals of calcipotriol monohydrate.

Fig.2b and 2c are graphs showing the results of analysis by differential scanning calorimetry (DSC) of two batches of nanocrystals calcipotriol monohydrate obtained by the method according to the present invention. DSC was carried out at 100ºC/min (Fig.2b) and at 100ºC/min (solid line), 300ºC/min (dotted line) and at 500ºC/min (dashed line) (Fig.2c). Several more thick if�Oia on the chart reflects exothermic phenomenon, emerging at about 8ºC and is reputed to be due to the crystallization of amorphous calcipotriol.

Fig.3 presents a graph showing the rate of release of calcipotriol from nanosuspensions of the present invention, compared with the rate of release of ointment Daivonex®. From the graph it appears that the rate of release is significantly higher from formulations in the form of nanosuspension than from Daivonex ointment®. "Cream in the form of nanosuspension is a cream in accordance with example 3. "Nanocosm. ointment. water" corresponds to A Composition of example 2 without glycerol, whereas "Nanocosm. ointment. glee ' is a Composition A of example 2.

Fig.4a presents a graph showing the penetration into the skin and the current through the skin of the two ointments in the form of nanosuspensions Compositions A and C of Example 2. "WSP ointment" is a Composition A, while the "salve Sonecon" is a Composition C.

Fig.4b presents a graph showing the penetration into the skin and the current through the skin of calcipotriol of ointments in the form of nanosuspensions Compositions A, C and D according to the invention, compared with the ointment Daivonex®. From the graph it appears that the penetration into viable skin from ointments in the form of nanosuspensions is comparable with the penetration of the ointment Daivonex®, although the current is significantly lower, resulting in less systemic exposure to Kalzip�of Tirol.

Fig.5 presents a graph showing the penetration into the skin and the current through the skin of calcipotriol of cream in the form of nanosuspension according to the invention, compared with Daivonex cream®. From the graph it appears that the penetration of calcipotriol of cream in the form of nanosuspension viable in the skin is significantly higher because of the cream in the form of nanosuspension than from Daivonex cream®.

Fig.6 schematically shows the activation of the gene encoding cathelicidin vitamin D3in human keratinocytes. The mechanism of activation of cathelicidin gene is used in biological analysis using reconstructed human epidermis (human keratinocytes, cultured in order to form the epidermal layers that are typical for human skin) to which are applied containing calcipotriol compositions according to the invention, activation of cathelicidin, as described in detail below in example 8.

DETAILED description of the INVENTION

Definitions

In the present context, the term "nanocrystal" is intended to indicate the crystalline particles of calcipotriol monohydrate, which are in the nanoscale range, i.e. with a diameter from 1 to 1000 nm. The nanocrystals preferably have particle size distributions that ≥90% of the nanocrystals have a particle size of from 100 to 900 nm, in particular from 200 to 600 nm.

�Ermin "nanosuspension" is intended to refer to nanocrystals, as defined above, suspended in the aqueous phase.

The term "distribution of particle size" is intended to denote the interval between the smallest and the largest crystal calcipotriol monohydrate, determined by dynamic light scattering (also known as photon correlation spectroscopy) using the apparatus Zetasizer Nano ZS or ZS90 (manufactured by Malvern Instruments, UK) according to the manufacturer's instructions. Dynamic light scattering determines the size of solid particles suspended in a fluid, by illuminating the particles with a laser and analyzing the intensity fluctuations in the scattered light as a result of Brownian motion of particles in a fluid. The intensity fluctuations are correlated with the particle size in that the larger particles move slower than smaller particles, i.e., the oscillation intensity is slower.

The term "amorphous" is intended to denote a solid substance without orderly arrangement of its molecules, i.e. the opposite of crystal.

"Calcipotriol" is an analogue of vitamin D formulas

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

The term "chemical�Skye stability" or "chemically stable" is intended to denote, the nanocrystals calcipotriol monohydrate not significantly decompose over time in a 24-EPI calcipotriol or other products of decomposition of calcipotriol in suspension or in a finished pharmaceutical product. In the case of the latter, "chemical resistance" indicates that no more than 10%, preferably not more than 6%, calcipotriol monohydrate decomposes during the shelf life of the product, usually 2 years at room temperature. Approximation of chemical resistance at room temperature get, exposing the nanocrystals or the composition containing them, accelerated stability studies 40ºC. If less than about 10% of the substance decomposed after 3 months storage at 40ºC, it is commonly accepted as an appropriate shelf-life of 2 years at room temperature.

The term "physical stability" or "physically stable" is intended to indicate that the nanocrystals calcipotriol monohydrate having substantially the form of crystals, identical control calcipotriol the monohydrate, which has not been subjected to bring to the nano range, according to the definition of the Romanov spectroscopy, i.e. it does not exhibit polymorphism as a result of bringing down to the nano range. In addition, "physical stability" indicates that the nanocrystals do not show agregatsiei the growth of crystals in the inventive suspensions or pharmaceutical compositions in which they are included.

The term "substantially non-aqueous" is intended to denote that the content of free water (unlike associated with crystal water) in lyophilized or subjected to spray-dried nanocrystals of calcipotriol monohydrate is less than about 2 wt.%, preferably less than about 1 wt.%, for example less than about 0.5 wt%. the nanocrystals. Similarly, the content of free water in "substantially nonaqueous" ointment composition is less than about 3 wt.%, preferably less than about 2 wt.%, for example, less than about 1%, or 0.5% of the mass. composition.

The term "solubilization capacity" is intended to indicate the ability of the solvent or mixture of solvents to dissolve the substance, expressed in terms of the amount required to fully solubilize substances.

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

The term "penetration through the skin" is intended to indicate the current of the active ingredient through the skin into the systemic circulation or, in the case studies,in vitrosuch as the studies described below in example 7, in the receptor fluid of the apparatus with the Franz cell, use�isobunnai in this experiment.

The term "biological activity" is intended to indicate the activity of a derivative or analog of vitamin D when applied to the skin in compositions according to the invention. The biological activity of the compositions is determined in the analysis ofin vitromeasurement of activation of the target genes encoding biological marker of cathelicidin in reconstructed human epidermis, including cultured human keratinocytes, as described in detail below in example 8.

Obtaining nanocrystals of calcipotriol monohydrate

In recent years there has been more research on nanocrystals or nanosuspensions therapeutically active ingredients as a way of ensuring improved the rate of dissolution of poorly soluble drugs. A large area of the surface of the nanocrystals provides a higher rate of dissolution upon introduction of the drug. This technology to date has mainly been used when enabling active ingredients in the composition of the formulations for oral or intravenous administration.

In the literature described several methods for producing nanocrystals of drugs. In a broad sense, methods can be divided into two categories, i.e., the grinding and homogenization under high pressure.

A publication�ation US No. 5145684 disclosed a method of producing crystalline nanoparticles by grinding in a ball mill for 4-5 days in the presence of surface modifiers, such as polyvinylpyrrolidone, polyvinyl alcohol, lecithin or other surfactant. It is likely that grinding in a ball mill of calcipotriol monohydrate in these conditions will lead or directly to chemical degradation of calcipotriol monohydrate, or to the formation of large quantities of amorphous calcipotriol that it would be unfavorable for sufficient stability during storage/shelf life of the pharmaceutical composition containing it, since the amorphous material is relatively more vulnerable to chemical degradation than crystalline material.

In the Sa publication No. 2375992 disclosed a method of producing particles of medicinal products with a particle size less than 5 microns, preferably less than 1 μm, homogenization under high pressure in a piston homogenizer with a pressure regulated through the gap, in an anhydrous medium at a temperature below 20 ° C, in particular below 0ºC. Bring to the nano range is the impact on micronized particles of the drug 10-20 cycles of homogenization under high pressure at 1500 bar. In the method according to the present invention, applicants use the aquatic environment to reduce the particle calcipotriol, as the use of anhydrous medium (liquid paraffin) did not lead to any meaningful reduction in the size of crystals calcipotriol mono�of ikata.

In the publication WO 2004/054549 disclosed local formulation of spironolactone nanoparticles in a cream base containing monoglycerides in water. Nanoparticles prepared using homogenization under high pressure in a piston homogenizer feed pressure through the adjustable gap.

It was found that homogenization under high pressure at 1500 bar for a few cycles not suitable for reducing particle calcipotriol monohydrate, since this procedure leads to the aggregation of crystals calcipotriol monohydrate even in the presence of a suitable surfactant.

In the publication WO 2008/058755 disclosed receipt of the nanocrystals cosmetically active substances by grinding with the use of Hollander or a ball mill, followed by homogenization under high pressure. Indicated that the combination of these two methods has advantages over a single homogenization under high pressure, since the latter requires many cycles of homogenization at high pressure (1500 bar). The combined method enables the use of only one homogenization cycle at low pressure to obtain particles of nanoscale.

In a preferred embodiment of the invention relates to a method for producing nanocrystals of calcipotriol monohydro�and the distribution of particle size in the range of 200-600 nm, according to the definition of dynamic light scattering, the method comprises the steps

(a) reducing the particle size of the crystalline calcipotriol monohydrate in an aqueous phase containing non-ionic polymeric surfactant in an amount of from about 1% to about 5 wt%. in the calculation of the aqueous phase with the formation of microparticles with a distribution of particle size in the range of about 5-20 μm and an average particle size of about 10 microns;

(b) effect on the suspension of step (a), the first cycle of homogenization under high pressure, comprising of about 300-800 bar for a period of time sufficient to obtain approximately 15-40% of crystals of calcipotriol monohydrate with the distribution of particle size in the range of 200-600 nm;

(c) effect on the slurry obtained in stage (b) second cycle of homogenization under high pressure, constituting approximately 800-1200 bar, for a time period sufficient to obtain approximately 40-80% of the crystals calcipotriol monohydrate with the distribution of particle size in the range of 200-600 nm;

(d) effect on the slurry obtained at the stage (c) third cycle of homogenization under high pressure, constituting approximately 1200-1700 bar, during a period of time sufficient to obtain about 90% or more of crystals of calcipotriol monohydrate with locatio�edelenyi particle size in the range of 200-600 nm; and

(e) optionally, separation of the obtained nanocrystals calcipotriol monohydrate from the aqueous phase.

In the final suspension (after stage (d)) is the number of nanocrystals calcipotriol monohydrate with the distribution of particle size in the range of 200-600 nm is preferably 95% or more.

A method according to the present invention differs from the method disclosed in the publication WO 2008/058755, by combining the initial stage of size reduction of the particles (a) with three consecutive cycles of homogenization under high pressure, and each cycle is carried out at increased pressure. This differs from the procedure disclosed in the publication WO 2008/058755, where for grinding in a ball mill the active ingredient should be one cycle of homogenization under high pressure at low pressure (such as 100 bar, see Examples 8 and 9), which results in the desired reduction in particle size. It was found that such a procedure is insufficient to provide a satisfactory particle size and size distribution of particles of crystals of calcipotriol monohydrate, i.e., only about 15-40% of the crystals would be within the desirable size distribution of the particles.

In addition, it was found that there is no need for carrying out the method according to the present invention with control of temperature, in contrast to the procedure, p�hidden in WO 2008/058755, where for two of the three active ingredients the temperature maintained below 20 ° C, and ideally from 0ºC to 5ºC. No need to use temperature control provides the advantage of a simplified procedure. However, surprisingly, in the method according to the present invention does not require temperature control because calcipotriol sensitive to temperature increases, one would expect that he would be subjected to chemical degradation without temperature control method of particle size reduction.

Embodiments of the invention

The suspension of the present invention may contain a nonionic, polymeric surfactant, which is added to prevent aggregation of the nanocrystals calcipotriol and/or to prevent the growth of crystals. A surfactant preferably is a kind of development that causes no significant solubilization of nanocrystals calcipotriol monohydrate, i.e. it must have a low ability to solubilize and can be favorably selected from the group consisting of surfactants in the form of poloxamers or polysorbates and simple C6-24alkyl esters of polyoxyethylene. Poloxamer can be selected from the group consisting of poloxamer 124, poloxamer 188, Paul�camera 237, poloxamer 338 and poloxamer 407. In particular, it was found that poloxamer 188 has a low solubilization capacity in relation to the solubilization of calcipotriol, and therefore is currently the preferred surfactants for use in nanosuspension of the present invention. When using Polysorbate as a surfactant can be selected from the group consisting of Polysorbate 20, Polysorbate 40, Polysorbate 60, Polysorbate 61, Polysorbate 80 and Polysorbate 81. Preferred in the present simple C6-24alkyl ether of polyoxyethylene is cetomacrogol 1000.

The amount of surfactant in the aqueous phase can be in the range from about 0.01% to about 5 wt%. suspension. In General, it is preferable that the amount of surfactant in said aqueous phase were in the range of about 0.6 to 1.2% of the mass. suspension.

Depending on the conditions applied during the reduction of particle size and homogenization under high pressure, the nanocrystals calcipotriol monohydrate present in the resulting suspension may have an average particle size of 200-350 nm, 350-400 nm or 400-500 nm, according to the definition of dynamic light scattering.

Nanosuspension can be lyophilized or subjected to spray drying to nanocrystals containing a surfactant on the surface. Lyophilized or subjected to raspy�rapidly drying the nanocrystals can then be used for inclusion in the nonaqueous composition.

Unexpectedly, it was found that the procedure of bringing to the nano range of the present invention generates only negligible amounts of amorphous calcipotriol. Specialist in the art it is well known that the presence of amorphous compounds makes the material unstable due to the lack of molecular arrangement in the crystal lattice, which subjects the material to chemical degradation or rearrangement of the crystal structure to another polymorphic form (see Chow et al., J. Pharm. Sci. 97(8), 2008, pp. 2855-2877). According to the definition by differential scanning calorimetry using the instrument Perkin-Elmer DSC 8500 in accordance with the manufacturer's instructions it was impossible to identify significant quantities of amorphous calcipotriol to the limit of detection of the instrument (approximately 5%), see Fig.2b.

In the method according to the present invention, the crystals of calcipotriol initially subjected to grinding or prednisolo in the aqueous phase using balls or beads with a diameter in the range of 1-4 mm, for example 2-3 mm. Balls or beads can be composed of glass or similar hard material, such as zirconium oxide. Grinding may suitably be carried out for 2-5 hours, for example 3 hours, at about 500-4000 rpm, such as about the speed 1000-3000 rpm, for example about 2000 rpm�n

Surfactants used for grinding may suitably be a non-ionic polymeric surfactant that is added to the aqueous phase, in an amount in the range from about 1.5 to about 3 wt%. suspensions, in particular about 2 wt%. suspension. The surfactant may preferably be selected from the group poloxamers or Polysorbate surfactants, as described above. Suspension is used directly in the subsequent stages of homogenization under high pressure, and especially favorable result on the distribution of particle size after homogenization under high pressure was obtained using poloxamer 188. It should be noted that due to the use in the production method according to the present invention washing equipment for grinding water at the stage (a) and washing with water equipment for homogenization under high pressure after stage (d), the surfactant concentration in the final slurry is in the range of about 0.6 to 1.2% of the mass. suspension.

Stage homogenization under high pressure (b)-(d) are conducted using a piston homogenizer with a pressure regulated through the gap, for example, a homogenizer Emulsiflex C3 (manufactured by Avestin), in accordance with the manufacturer's instructions.

It was found that in order to nano-sized crystals of calcipotriol monohydrate favorable�TNO, to the first cycle of homogenization under high pressure stage (b) was carried out at a pressure of approximately 500-650 bar. The time required to obtain 15-40% of the nanocrystals calcipotriol monohydrate with desired distribution of particle size is 7-15 minutes, for example 8-12 minutes, accounting for such period as of approximately 10 minutes.

The second cycle of homogenization under high pressure stage (c) may suitably be carried out at a pressure of about 1000-1100 bar. The time required to obtain 40-80% of the nanocrystals calcipotriol monohydrate with desired distribution of particle size is in the range of 7-15 minutes, for example 8-12 minutes, accounting for such period as of approximately 10 minutes.

The third cycle of homogenization under high pressure stage (d) may suitably be carried out at a pressure of about 1400-1500 bar. The time required to obtain 90% or more of nanocrystals calcipotriol monohydrate with desired distribution of particle size is in the range of 7-15 minutes, for example 8-12 minutes, accounting for such period as of approximately 10 minutes.

If the nanocrystals calcipotriol monohydrate intended for inclusion in the nonaqueous preparative form, they can appropriately be subjected to lyophilization or spray drying (until the water content (free water) less than about 2 wt.%, this makmanee than about 1%, or less than about 0.5 wt%. nanocrystals.

The nanocrystals calcipotriol monohydrate or suspension containing the nanocrystals can be included in a pharmaceutical composition containing a pharmaceutically acceptable carrier which is compatible with the active ingredient.

When mixed with pharmaceutically acceptable excipients to obtain a composition as described below, the amount of the nonionic polymeric surfactant is preferably in the range of about 0.03 to 0.06% of the mass. composition.

In one embodiment of the composition according to the present invention is an ointment. According to the modern classification of FDA (Administration food and drug administration) ointment is a semisolid dosage form which can contain water and volatile substances in an amount up to 20 wt%. and which contains more than 50% of the mass. hydrocarbons, waxes, or polyols in the media. Thus, in accordance with the invention, the ointment can be a composition of water-in-oil, and in this case the very nanosuspension can be added to the lipophilic components of the composition so that the composition contained up to 10 wt%. or preferably to 5 wt%. the water phase. Alternatively, the composition may be a non-aqueous ointment, which contains less than about 2%, preferably less than 1% with�rim water by weight of the composition.

Ointment carrier may suitably contain a paraffin selected from paraffins consisting of hydrocarbons with lengths of chains in the range of C5-60and their mixtures. Frequently used ointment carrier is a petrolatum or white soft paraffin, which is composed of hydrocarbons with different chain length, reaching a peak approximately at C40-44or a mixture of petrolatum and liquid paraffin(consisting of hydrocarbons of different chain length, reaching a peak approximately at C28-40). Although petrolatum provides occlusion of the treated skin surface, reducing transdermal loss of water and potentiating therapeutic effect of the active ingredient in the composition, it tends to cause a feeling of oiliness and/or stickiness, which persists a long time after application, and it is not easy distributed over the surface of the skin. Therefore, it may be preferable to use paraffins consisting of hydrocarbons of a somewhat lower chain length, such as paraffin wax consisting of hydrocarbons with lengths of chains, reaching a peak for C14-16, C18-22, C20-22, C20-26or their mixtures. It was found that these waxes are more cosmetically acceptable in that they are less sticky and/or oily after application and easier distributed across the surface of the skin. Poet� expected they will lead to better patient compliance with the prescribed treatment. Suitable waxes of this type are issued by Sonneborn and sold under the trade name Sonecon, for example, Sonecon CM, Sonecon DM1, Sonecon DM2 and Sonecon HV. These paraffins are described in more detail and described in WO 2008/141078, which is incorporated into this description by reference. (Paraffin hydrocarbon composition was determined by gas chromatography.)

To impart the desired viscosity of the composition of the present invention, it may be appropriate to include lipophilic increasing the viscosity of the ingredient, such as wax. The wax may be a mineral wax composed of a mixture of hydrocarbons with high molecular weight, e.g., saturated C35-70alkanes, such as microcrystalline wax. Alternatively, the wax may be a vegetable or animal wax, such as ester C14-32fatty acids and C14-32fatty alcohols, such as beeswax. The number of increasing the viscosity of ingredient may vary in accordance with imparting viscosity force ingredient, but may generally be in the range of about 1-20 wt%. composition. When increasing the viscosity of ingredient is a microcrystalline wax, it is usually present in quantities� in the range of about 5-15 wt.%, for example, about 10 wt%. composition.

To maintain sufficient physical stability of the composition, in particular to avoid split in it water and lipid phases, it may be preferable to the inclusion of an emulsifier water-in-oil with HLB value (water-lipid balance) 3-8. Examples of such emulsifiers are the simple C8-22alkyl esters of polyoxyethylene, for example, stearyl simple ether of polyoxyethylene, simple cetyl ether polyoxyethylene, simple alerby broadcast polyoxyethylene or simple lauryl ether of polyoxyethylene. The amount of emulsifier is typically in the range 2-10% wt./mass. composition.

The composition may optionally contain a softening agent, which may act to soften the thickened epidermis of psoriatic plaques. Suitable softening agent for inclusion in the composition of the present invention may be a silicone wax or a volatile silicone oil, because it was additionally found that the presence of silicone promotes the penetration of calcipotriol in the skin. It was also discovered that compositions comprising silicone, lead to less skin irritation. Suitable silicone oils for inclusion in the composition of the present invention may be selected from cyclomethicone and Dimethicone. The amount of the silica�the new oil included in the composition of the present invention, is typically in the range from about 1 to about 10 wt.%, for example, about 5 wt%. composition.

It is believed that the presence in the ointment Daivonex® propylene glycol is the main factor causing skin irritation experienced by many patients. However, it was found that calcipotriol may in some patients cause slight irritation (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 favorable for the inclusion in the composition of the present invention compounds against irritation, such as glycerol, sorbitol, sucrose, saccharin, menthol, eucalyptol or nicotinamide. Glycerol has been described as a substance that can protect the skin from irritants (J. Bettinger et al., Dermatology 197, 1998, pp. 18-24), and was found by applicants to reduce the release of interleukin IL-1α-dependent way: for example, it was found that the presence of 15 wt%. of glycerol to ointments calcipotriol led to a significantly lower level of release of IL-1α than the inclusion of 10 wt%. glycerol, which, in turn, led to a significantly lower level of release of IL-1α than the inclusion of 5 wt%. of glycerol.

However, in addition to the effect against irritation, unexpectedly, it was found that glycerol is able sweat�to neeravari biological activity of calcipotriol in that, as observed, the expression of cathelicidin (in the analysis described below in example 4) increases low amount of glycerol in the composition (i.e., cathelicidin more expressed when the amount of glycerol is 5% wt., than when the amount of glycerol is 10% or 15%): this implies that the inclusion of glycerol, a balance must be achieved between favourable effect against irritation and potentiating a favorable effect. Applicants have found that the inclusion of about 5-10% of the mass. of glycerol in the presence of the composition leads to a significant effect against irritation, as well as to a significant potenzirovania biological activity of calcipotriol.

It is known that calcipotriol is a substance, which is highly sensitive to acid conditions (pH below about 7.0 in aqueous compositions or acid reacting substances in non-aqueous compositions) that contribute to the rapid destruction of calcipotriol. To ensure adequate chemical stability of the substance during the storage period of the composition may be desirable inclusion compound capable of neutralizing acidic impurities which may be present in one or more of excipients composition and chemical are harmful to the sustainability of calcipotriol. EU�and composition is an aqueous, the acid neutralizing compound can be selected from a buffer, such as phosphate buffer, which may be included in amounts of 0.025-0,065% of the mass. composition. On the other hand, if the composition is non-aqueous, acid neutralizing compound may advantageously be a amine, such as triethanolamine, trometamol, monoethanolamine or diethanolamine, are included in the composition in an amount of 0.1-2 wt%. composition.

In another embodiment of the composition according to the present invention is a cream that may contain features that are similar to the ointment, but which is usually an emulsion of oil-in-water containing significant amounts of water.

In a specific embodiment of the composition according to the present invention contains:

0,003-0,008% wt./mass. calcipotriol monohydrate

2-8% wt./mass. simple stearyl ether of polyoxyethylene

2-10% wt./mass. water

0,001-0,005% wt./mass. poloxamer 188

70-90% of the mass./mass. paraffin medium.

The composition of the present invention may also contain other components commonly used in dermal formulations, e.g. antioxidants (e.g. alpha-tocopherol), preservatives, sodium edetate, pigments, softening the skin agents, healing the skin agents and skin conditioning agents such to�to urea, allantoin or bisabolol, see the Manual CTFA (Association of cosmetics, hygiene products and flavorings) Cosmetic Ingredients Handbook, 2ndEd., 1992.

A composition according to the invention can be used in the treatment of psoriasis, sebopsoriasis, Palmar-plantar pustules, dermatitis, ichtyosis, rosacea and acne and related skin diseases by local application of an effective amount of a composition in accordance with the invention, in need of such treatment of a patient. The method preferably includes the local application once or twice per day a therapeutically sufficient dose of the indicated composition. For this arrangement in accordance with the invention preferably contains about 0.001 to 0.5 mg/g, preferably about from 0.002-0.25 mg/g, in particular from 0.005 to 0.05 mg/g of nanocrystals calcipotriol monohydrate. It is envisaged that a composition according to the present invention may advantageously be used for the maintenance treatment of these skin diseases, i.e. ongoing treatment after the disappearance of visible symptoms, to delay the recurrence of symptoms.

To ensure a more effective treatment of psoriasis and other skin diseases in the acute phase, it may be desirable to add one or more additional therapeutically active ingredients in the composition Examples of such additional active ingredients include without limitation anti-inflammatory medicines, like corticosteroids, such as betamethasone and its esters, for example, an ester valerate or dipropionate, clobetasol or its esters such as the propionate, hydrocortisone or their esters, such as acetate; non-steroidal anti-inflammatory drugs such as naproxen, indomethacin, diclofenac, ibuprofen, dexibuprofen, Ketoprofen, flurbiprofen, piroxicam, lornoxicam or nabumeton, inhibitors of phosphodiesterase 4 (e.g., compounds disclosed in WO 2008/077404, WO 2008/104175, WO 2008/128538 or WO 2010/069322) or inhibitors of p38 MAP-kinase (for example, joints, disclosed in WO 2005/009940 or WO 2006/063585).

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

EXAMPLES

Example 1

Obtaining nanocrystals of calcipotriol monohydrate

4 g poloxamer 188 was dissolved in 196 ml of laboratory water with stirring and the pH was adjusted to 8.5 by adding appropriate amount of NaOH.

3.5 g of glass balls with a diameter of 2 mm were weighed in two bottles provided with screw cap. To each vial was added 0.035 g of calcipotriol monohydrate, then in each vial was added 1.05 g of 2% solution poloxamer 188. Calcipotriol monohydrate crushed Stra�ywaniem on an orbital shaker (VXR Basic IKA Vibrax) at 2000 rpm.

After grinding bottles and glass balls, used for grinding, washed with 24.0 g of laboratory water at pH 8.5 and suspension of calcipotriol monohydrate was poured in the bottle with a blue lid. The suspension was transferred into a high pressure homogenizer Emulsiflex C3 (Avestin), and a bottle with a blue lid was washed with 4,9 g laboratory water at pH 8.5. Homogenization under high pressure were carried out at 500 bar for 10 minutes at 1000 bar for 10 minutes and 1400 bar within 10 minutes. After homogenization under high pressure cylinder apparatus Emulsiflex washed 4,9 g laboratory water at pH 8.5, and then determined the distribution of particle size by dynamic light scattering with Zetasizer Nano ZS90, wherein the particle size distributions were in the range of 200-600 nm, and the average particle size was in the range 350-400 nm.

By using Raman spectroscopy, comparing the Raman spectrum of the nanocrystals with the Raman spectrum of calcipotriol monohydrate that has not been subjected to bring to the nano range, it was determined that the obtained nanocrystals are calcipotriol monohydrate.

The amount of amorphous calcipotriol generated in this way were determined on two batches of nanocrystals calcipotriol obtained by the method using DSC analysis �ri heating rate of 100ºC, 300ºC and 500ºC/min in the atmosphere of N2. The instrument used for analysis was a Perkin Elmer DSC 8500.

The results are shown in Fig.2b and 2c, showing the exothermic phenomenon beginning at about 8ºC. It is considered highly probable that the exothermic phenomenon caused by crystallization of amorphous calcipotriol. It appears that the amount of heat emitted during the process of crystallization, very little, in fact, very close to the limit of detection. Since the amount of heat emitted during the process of crystallization is proportional to the amount of amorphous compounds present in the sample, the applicants concluded that the two parties were present only small amounts of amorphous calcipotriol.

Example 2

Ointments containing nanocrystals of calcipotriol monohydrate

Ointment compositions shown below in table 1, were obtained by mixing the ingredients of the lipid phase (hydrocarbons + stearyl simple ether of polyoxyethylene(2) + α-tocopherol) when heated to 80-85ºC and slow stirring. The aqueous phase was prepared by dissolving denetria of edetate and denetria of dihydrogen phosphate in an appropriate amount of water nanosuspension of calcipotriol monohydrate (obtained as described in example 1), with the content of calcipotriol monohydrate, brought to 50 mcg/g. � suspension was added glycerol by mixing and heating to 35-40ºC and pH of the mixture was adjusted to 8.5, in accordance with the expediency of 1N HCl or NaOH.

The aqueous phase was added to the lipid phase while beating for 30 min, after which the resulting ointment is slowly cooled to a temperature below 32 C, it was filled aluminum tubes and kept at room temperature.

Table 1
Ingredient (mg/g)Conn. AConn. BConn. CConn. DConn. EConn. F
The nanocrystals calcipotriol monohydrate0,050,050,050,050,050,05
Disodium dihydrogen phosphate0,260,260,260,260,260,26
Paraffin, liquid5050505050 50
Simple stearyl ether polyoxyethylene(2)505050505050
Disodium edetate0,0650,0650,0650,0650,0650,065
Fully racemic α-tocopherol0,020,020,020,020,020,02
Glycerol100-100100100100
Water purified5015050505050
Poloxamer 1880,030,030,03 0,03--
Poloxamer 407----0,03-
Polysorbate 80-----0,03
Paraffin, white soft749,6--699,6749,6749,6
Cyclomethicone---50--
White Petrolatum Jelly (Sonecon DM1)-649,6649,6---
Microcrystalline wax (Multiwax 180 MH)-100 100---

The compositions were tested for chemical stability for 3 months at 40ºC/75% RH (relative humidity). The results show satisfactory stability of calcipotriol in test conditions.

Example 3

Cream containing nanocrystals of calcipotriol monohydrate

Cream compositions shown below in table 2, were obtained by melting of cetomacrogol 1000, cetosteatil alcohol, liquid paraffin at 80ºC. The aqueous phase was prepared by dissolving of disodium dihydrogen phosphate and chlorethylene chloride in purified water at 80ºC. To a solution of glycerol was added while mixing and the pH of the mixture was adjusted to 8.5, in accordance with the expediency of 1N HCl or NaOH.

The aqueous phase was mixed with the lipid phase during homogenization and cooled to 55ºC. The remaining water was added with vigorous stirring and the resulting cream was cooled to 25 ° C while stirring with a slow speed.

The corresponding amount of nanosuspension of calcipotriol monohydrate (obtained as described in example 1) to increase the content of calcipotriol monohydrate 50 mcg/g was added to the cream by mixing for 30 minutes at <30ºC. Received cream filled tubes and stored until further use.

Table 2
Ingredient (mg/g)Connection GConnection HThe connection I
The nanocrystals calcipotriol monohydrate0,050,050,05
Cetomacrogol 1000303030
Cetosteatil alcohol606060
Chlorethylamines chloride0,50,50,5
Glycerol303030
Disodium dihydrogen phosphate222
Poloxamer 1880,03--
Poloxamer 407-0,3 -
Polysorbate 80--0,03
Paraffin, liquid505050
Paraffin, white soft170170170
Water purifiedTo 1 gTo 1 gTo 1 g

Cream compositions were tested for chemical stability for 3 months at 40ºC/75% RH. The results show satisfactory stability of calcipotriol in test conditions.

Example 4

Non-aqueous ointment containing nanocrystals of calcipotriol monohydrate

Nanosuspension of calcipotriol monohydrate, obtained as described in example 1 was subjected to lyophilization overnight. Lyophilized, essentially anhydrous nanocrystals was used to obtain ointments dispersion of nanocrystals in liquid paraffin and white soft paraffin to a variance.

The composition is non-aqueous ointments presented in table 3 below.

Table 3
IngredientContent
The nanocrystals calcipotriol monohydrate50 mcg
Paraffin, liquid50 mg
Paraffin, white soft750 mg
Poloxamer 1880.05 mg

Non-aqueous ointment were tested for chemical stability for 3 months at 40ºC/75% RH. The results show satisfactory stability of calcipotriol in test conditions.

Example 6

The release of nanosuspension compositions compared with Daivonex ointment®

The release ofin vitrocalcipotriol of the compositions described in examples 1 and 2 was determined in a diffusion cell made of plexiglass using a membrane Spectra/Porv 6 for the separation of receptor and donor chambers (n=6 cells per batch). Determined the release of calcipotriol in the receptor phase, consisting of 0.04 M isotonic phosphate buffer, pH 7.4 and isopropanol (70:30 vol./vol.). The samples were analyzed by HPLC/UV.

The results are presented below in Fig.3, show that the rate of release of calcipotriol from nanosuspension ointments and creams for this�th invention is significantly higher than the rate of release of ointment Daivonex®.

The results presented in Fig.3, show that the rate of release is significantly higher from nanosuspension formulations than from Daivonex ointment®.

Example 7

The study of skin penetrationin vitro

To study the penetration of calcipotriol into the skin and through the skin of the compositions according to the invention, an experiment was conducted diffusion through the skin. This study used a full-thickness skin from the ears of pigs. The ears were kept frozen at-18ºC before use. A day before the experiment, the ears were placed in a refrigerator (5±3ºC) for slow thawing. On the day of the experiment, the hair was removed using a small animal clipper. The skin cleared of subcutaneous fat using a scalpel, and two pieces of skin were cut off from each ear and placed in the diffusion cell Franz in a balanced manner.

Static diffusion cell of the type Franz with an available diffusion area 3,14 cm2and receptor volumes in the range from 8.6 to 11.1 ml was used essentially in the same manner as described in T. J. Franz, "The finite dose technique as a valid in vitro model for the study of percutaneous absorption in man", in Current Problems in Dermatology, 1978, J. W. H. Mall (Ed.), Karger, Basel, pp. 58-68. Specific volume was measured and recorded for each cell. A magnetic bar was placed in the receptor compartment of each cells the correct l�. After you install the skin each receptor chamber was filled with physiological saline solution (35ºC) for skin hydration. The cells were placed in a thermally controlled water bath, which was placed on a magnetic stirrer set at a speed of 400 Rev/min. the temperature of the circulating water in the baths was maintained at 35±1ºC, that provided the temperature of about 32ºC on the surface of the skin. After one hour the saline solution was replaced with receptor medium of 0.04 M isotonic phosphate buffer with a pH of 7.4 (35ºC) containing 4% bovine serum albumin. The terms of the dip maintained at all times during the study period, i.e. the concentration of active compounds in the receptor medium was below 10%, the solubility of compounds in the environment.

Skin penetrationin vitroeach of the tested compositions were tested in the 6 repetitions (i.e., n=6). Each test composition was applied to the skin membrane at 0 hours in the intended dose of 4 mg/cm2. For application used a glass spatula and a residual amount of the composition was determined in order to obtain the number of songs that really applied to the skin.

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

The stratum corneum took the charger�the group from the ribbon 10 times using tape (D-Squame® (diameter 22 mm, CuDerm Corp., Dallas, Texas, USA). Each strip of tape was applied to the test area using the standard pressure for 5 seconds, and removed from the area to be tested in one continuous motion blurred. For each re-tape, the direction of separation were varied. Then similarly from the skin samples were taken for viable epidermis and dermis.

Took and analyzed samples (1 ml) of the receptor fluid remaining in the diffusion cell.

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

The results are presented in Fig.4a, which shows the penetration into viable skin composition of A and C using two different paraffin carriers, and Fig.4b, which shows that the penetration into viable skin from resuspension ointments comparable with the penetration of the ointment Daivonex®, although the current is significantly lower, resulting in less systemic effects of calcipotriol.

Other results shown in Fig.5, which is a graph showing that the penetration of calcipotriol from composition G in viable skin is significantly higher from nanosuspension cream than cream Daivonex®.

Example 8

The biological activity of compositions

As shown below in Fig.6, �utilizedin is an antimicrobial peptide, expressed in human keratinocytes. Strong expression of cathelicidin causes infection of the skin or destruction of the skin barrier. In psoriasis level increases cathelicidin in skin lesions of patients with psoriasis. It was found that the expression of the gene encoding cathelicidin, can be caused by vitamin D3or analogues of vitamin D, such as calcipotriol (see publications Wang TT 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), 22 July 2009, e6340) by binding with the receptor of vitamin D. These data were used to develop the analysis, which capture and biological activity of calcipotriol in human keratinocytes of the tested compositions was determined by measuring the level of induction of the gene encoding cathelicidin.

In this analysis, containing the nanocrystals calcipotriol monohydrate cream obtained as described above in example 3 (composition G) was applied topically in three repetitions on reconstructed human epidermis consisting of normal human keratinocytes cultured for 12 days on polycarbonate filters with an area of 0.5 cm2(produced by SkinEthic® Laboratories, Nice, France) in an amount of 10 μl. The fabric is processed within one or two days in the presence of cytokines IL-7 (20 ng/ml), IL-22 (20 ng/ml) and TNF-α (5 ng/ml), followed by the separation of the epidermis from the polycarbonate filter and instant freezing in liquid nitrogen. RNA was extracted from cells and cDNA was synthesized by conventional procedures. Then performed quantitative PCR (qPCR) using the following assays from Applied Biosystems: CAMP Hs0018038_m1 and GAPDH Hs99999905_m1. The expression levels of cathelicidin were normalized to GAPDH and relative quantification was performed by comparison with ointment and cream Daivonex®.

The results are presented below in table 4.

Table 4
CompositionDay 1 activation12nd day activation1
Ointment Daivonex®2,34,1
Cream Daivonex®1,02,2
Composition G2,54,7
1The multiplicity of activation relative to the Daivonex cream®, 1-day.

The results presented in table 4 show that the application of the Composition G resulted in activation of the target genes, similar Akti�ation, the resulting ointment Daivonex®, although activation of the target genes was approximately double activation cream Daivonex®. Thus, the results indicate the efficiency that is as high as the activity achieved with ointment Daivonex®, obtained from preparative form, which does not contain propylene glycol, which has a more favorable cosmetic properties.

Compositions A, C and D obtained as described above in example 2 was applied topically in three repetitions on reconstructed human epidermis consisting of normal human keratinocytes cultured for 12 days on polycarbonate filters with an area of 0.5 cm2(produced by SkinEthic® Laboratories, Nice, France) in an amount of 10 μl. The fabric was treated for two days followed by separation of the epidermis from the polycarbonate filter and instant freezing in liquid nitrogen. RNA was extracted from cells and cDNA was synthesized by conventional procedures. Then was fulfilled qPCR using the following assays from Applied Biosystems: CAMP Hs0018038_m1 and GAPDH Hs99999905_m1. The expression levels of cathelicidin were normalized to GAPDH and relative quantification was performed by comparison with ointment and cream Daivonex®.

The results are presented in table 5 below.

Table 5
CompositionThe multiplicity activation1
Ointment Daivonex®1,0
Composition A2,6/2,1
Composition C1,3/4,1
Composition D4,1/6,8
1About ointment Daivonex®.

The results presented in table 5 show that the compositions according to the invention lead to a higher activation of the target genes, i.e. they have a higher biological activity than the ointment on the market.

Example 5

A study of the local application of the mini-pigs

Tolerability of the compositions A, C and D of example 2, the local application was evaluated by daily application to the skin mini pigs for 4 weeks. Ointment Daivonex® was used for comparison. Every day the animals were exposed to test products for 8 hours.

The study was performed in 10 female mini-pigs Gottingen SPF. Each animal was 6 areas of application, and it received 250 mg of the tested formulations at the site of application. Clinical signs were recorded daily and skin reactions at the sites of application were evaluated� in points once a day before I applied it, in addition, on the day of autopsy, in respect of erythema and edema. Feed intake was recorded daily and body weight weekly. At the end of the treatment period was performed macroscopic pathology all animals and skin samples were taken for histopathological studies.

The results show that during the study there were no side associated with treatment of clinical signs, although there was a skin reaction (erythema) of 1-2 degrees. Except for compositions A, erythema were less pronounced than erythema observed for ointment Daivonex®. The results indicate that the composition of the invention can better be carried by patients than ointment Daivonex®.

1. Suspension for the treatment of psoriasis, comprising calcipotriol monohydrate in the form of nanocrystals with a size distribution of particles in the range of 200-600 nm according to the definition of dynamic light scattering, which is dispersed in an aqueous phase comprising non-ionic, polymeric surfactant selected from the group consisting of surfactants in the form of poloxamers or polysorbates, in an amount of 0.01-5 wt.% in the calculation of the slurry to prevent the formation of aggregates and/or crystal growth of nanocrystals calcipotriol monohydrate, wherein the nanocrystals calcipotriol monohydrate obtained in suspension by processing the slurry by the method comprising stud�and:
(a) reducing the particle size of the crystalline calcipotriol monohydrate in an aqueous phase containing non-ionic, polymeric surfactant in an amount in the range from about 1 to about 5 wt.% in the calculation of the aqueous phase, with the formation of microparticles with a distribution of particle size in the range of about 5-20 μm and an average particle size of about 10 microns;
(b) effect on the suspension of step (a), the first cycle of homogenization under high pressure, comprising of about 300-800 bar for 7-15 minutes to obtain approximately 15-40% of crystals of calcipotriol monohydrate with the distribution of particle size in the range of 200-600 nm;
(c) effect on the slurry of step (b), the second cycle of homogenization under high pressure, constituting approximately 800-1200 bar, for 7-15 minutes to obtain approximately 40-80% of the crystals calcipotriol monohydrate with the distribution of particle size in the range of 200-600 nm; and
(d) effect on the slurry of step (c), the third cycle of homogenization under high pressure, constituting approximately 1200-1700 bar, for 7-15 minutes to obtain a suspension of approximately 90% or more of crystals of calcipotriol monohydrate with the distribution of particle size in the range of 200-600 nm.

2. Suspension according to claim 1, where poloxamer selected from the group consisting of poloxamer 124, Paul�camera 188, poloxamer 237, poloxamer 338 and poloxamer 407.

3. Suspension according to claim 2, where the surfactant is poloxamer 188.

4. Suspension according to claim 1, wherein the Polysorbate is selected from the group consisting of Polysorbate 20, Polysorbate 40, Polysorbate 60, Polysorbate 61, Polysorbate 80 and Polysorbate 81.

5. Suspension according to any one of claims. 1-4, where the amount of surfactant in said aqueous phase is in the range from about 0.6 to about 1.2 wt.% in the calculation of the suspension.

6. Suspension of claim 1, wherein the nanocrystals calcipotriol monohydrate has an average particle size of 200-350 nm, 350-400 nm or 400-500 nm according to the definition of dynamic light scattering.

7. A method of obtaining a suspension of nanocrystals of calcipotriol monohydrate according to any one of claims. 1-6, the method comprises the steps:
(a) reducing the particle size of the crystalline calcipotriol monohydrate in an aqueous phase containing a nonionic, polymeric surfactant selected from the group consisting of surfactants in the form of poloxamers or polysorbates, in an amount in the range from about 1 to about 5 wt.% in the calculation of the aqueous phase, with the formation of microparticles with a distribution of particle size in the range of about 5-20 μm and an average particle size of about 10 microns;
(b) effect on the suspension of step (a), the first cycle of homogenization under high pressure, comprising about 300800 bar, for 7-15 minutes to obtain approximately 15-40% of crystals of calcipotriol monohydrate with the distribution of particle size in the range of 200-600 nm;
(c) effect on the slurry of step (b), the second cycle of homogenization under high pressure, constituting approximately 800-1200 bar, for 7-15 minutes to obtain approximately 40-80% of the crystals calcipotriol monohydrate with the distribution of particle size in the range of 200-600 nm;
(d) effect on the slurry of step (c), the third cycle of homogenization under high pressure, constituting approximately 1200-1700 bar, for 7-15 minutes to obtain a suspension of approximately 90% or more of crystals of calcipotriol monohydrate with the distribution of particle size in the range of 200-600 nm.

8. A method according to claim 7, where the stage of size reduction of the particles (a) is carried out by grinding in a ball mill in a wet state using balls or beads with a diameter in the range of 1-4 mm, such as 1.5-2.5 mm or 2-3 mm.

9. A method according to claim 7, where poloxamer selected from the group consisting of poloxamer 124, poloxamer 188, poloxamer 237, poloxamer 338 and poloxamer 407.

10. A method according to claim 9, where the surfactant is poloxamer 188.

11. A method according to claim 7, where the Polysorbate is selected from the group consisting of Polysorbate 20, Polysorbate 40, Polysorbate 60, Polysorbate 61, Polysorbate 80 and Polysorbate 81.

12. A method according to claim 7, where �AB is added in stage (a) in an amount in the range from about 1.5 to about 3 wt.% suspension, in particular about 2 wt.% suspension.

13. A method according to claim 7, where the first cycle of homogenization under high pressure stage (b) is conducted at a pressure of approximately 500-650 bar.

14. A method according to claim 7, wherein the second cycle of homogenization under high pressure stage (c) is conducted at a pressure of about 1000-1100 bar.

15. A method according to claim 7, where the third cycle of homogenization under high pressure stage (d) is conducted at a pressure of about 1400-1500 bar.

16. A method according to claim 7, where the stage homogenization under high pressure (b)-(d) are conducted using a piston homogenizer with a pressure regulated through the gap.

17. Pharmaceutical composition intended for the treatment of psoriasis containing nanocrystals of calcipotriol monohydrate according to any one of claims. 1-6 and a pharmaceutically acceptable carrier.

18. A composition according to claim 17, where the amount of the nonionic polymeric surfactant is in the range of about 0.03 to 0.06 wt.% composition.

19. A composition according to claim 17, where the media includes at least one paraffin selected from paraffins consisting of hydrocarbons with lengths of chain from C5to C60and the length of the chains reach a peak for C14-16, C18-22, C20-22, C20-26, C28-40and C40-44or their mixtures.

20. A composition according to claim 17, further comprising an emulsifier water-in-oil selected from a simple C8-22alkyl esters of�of oxyethylene, for example stearyl simple ether of polyoxyethylene, simple catelouge ether of polyoxyethylene or simple lauryl ether of polyoxyethylene.

21. A composition according to claim 17, further comprising an ingredient that increases the viscosity.

22. A composition according to claim 21, where increasing the viscosity of ingredient is a wax.

23. A composition according to claim 17, further comprising a silicone wax or a volatile silicone oil.

24. A composition according to claim 23, where the volatile silicone oil is cyclomethicone or Dimethicone.

25. A composition according to claim 17, further comprising interstriae connection.

26. A composition according to claim 25, where interstriae the compound is a glycerol, sorbitol, sucrose, saccharin, nicotinamide, menthol or eucalyptol.

27. A composition according to claim 17, further comprising a compound capable of neutralizing acidic impurities, harmful chemical resistance of calcipotriol monohydrate in the composition.

28. A composition according to claim 27, where the specified connection is an amine, such as triethanolamine, trometamol, monoethanolamine or diethanolamine.

29. A composition according to claim 17, which is an ointment.

30. A composition according to claim 29, which represents an essentially anhydrous ointment.

31. A composition according to claim 17, which is a cream.

32. Songs�I according to claim 17, containing approximately 0,001-0,5 mg/g, preferably about from 0.002-0.25 mg/g, in particular from 0.005 to 0.05 mg/g of nanocrystals calcipotriol monohydrate.

33. A composition according to claim 17, further comprising one or more additional therapeutically active ingredients.

34. A composition according to claim 33, wherein additional active ingredients selected from the group consisting of anti-inflammatory drugs such as corticosteroids, such as betamethasone and its esters, for example an ester valerate or dipropionate, clobetasol or its esters such as the propionate, hydrocortisone or their esters, such as acetate; non-steroidal anti-inflammatory drugs such as naproxen, indomethacin, diclofenac, ibuprofen, dexibuprofen, Ketoprofen, flurbiprofen, piroxicam, lornoxicam or nabumeton, inhibitors of phosphodiesterase 4 inhibitors or p38 MAP kinase.



 

Same patents:

FIELD: electricity.

SUBSTANCE: invention is related to electrochemical installation intended to shape nanosized coating and may be used in semiconductor and electronics industry. The installation contains a computer, a controller and manipulator 1 mounted at the rack 2 rotatable around vertical axis and equipped with holder 3 for a processed sample 4. Around the manipulator 1 rack there are electrochemical cells 5 with electrodes connected to one pole of current source. The sample 4 submerged to electrochemical cells is connected to the other pole of current source. Holder 3 is installed so that it can be moved in regard to manipulator 1, and at that sample 4 in downwardmost position of holder 3 is placed in one of electrochemical cells. One of electrochemical cells is made as measuring cell 7 to control parameters of the processed sample 4. The installation is equipped with tube-type furnace 8 intended for thermal processing of the sample.

EFFECT: potential determining and setting of the required parameters for obtained nanomaterial against absolute value and conditions of their change.

4 dwg

FIELD: physics.

SUBSTANCE: proposed shutter comprises locally smelting or evaporating mirror metal film located in focal area of the lens and secured by translucent substrate. On radiation side said substrate includes also the ply of translucent liquid of solid sol with nanoparticles in size smaller than radiation wavelength. Mirror film is arranged on said substrate on radiation side or opposite side.

EFFECT: lower threshold of shutter operation.

4 dwg

FIELD: chemistry.

SUBSTANCE: method includes treating the surface of crystalline silicon by electrochemical etching in hydrofluoric acid solution with concentration of 20-30% while supplying current with surface density of 750-1000 mA/cm2 for 5-30 s to obtain hydrophobic silicon or supplying current with surface density of not more than 650 mA/cm2 for 5-30 s to obtain hydrophilic silicon.

EFFECT: method enables to obtain a surface with multimodal nano- or microporosity in a single step.

4 dwg

FIELD: chemistry.

SUBSTANCE: invention relates to a method of producing electret fine particles or coarse powder. The method of producing electret fine particles includes steps of emulsifying a fluorine-containing material which contains a vinylidene fluoride-hexafluoropropylene-tetrafluoroethylene terpolymer in a liquid which is immiscible with the fluorine-containing material to obtain emulsified particles or microcapsule particles and subjecting the emulsified particles or microcapsule particle to irradiation with an electron beam, radioactive radiation, or corona discharge treatment. A version of a method of producing a coarse powder is also disclosed.

EFFECT: excellent electrophoretic properties.

8 cl, 4 tbl, 5 dwg, 24 ex

FIELD: chemistry.

SUBSTANCE: method for producing a nanocomposite anti-reflective coating involves preparing a liquid composition containing a solution of carboxyl-containing acrylate copolymer mixed with solvents of various volatility and silver nanoparticles produced directly in the solution by reducing with solvent-borne aldehyde by UV radiation. The prepared nanocomposition covers a solid-state underlying surface; then it is dried.

EFFECT: simplifying technology, reducing the aggregating effect on a disperse phase of the suspension, increasing the coating adhesion.

1 tbl, 4 ex

FIELD: chemistry.

SUBSTANCE: disclosed is a petroleum lubricating oil additive which contains ultrafine diamond powder treated with fluorine-containing oligomers with average molecular weight of 2200-4200 units and a general structural formula Rf-CF3 or Rf-COOH, where Rf is a fluorine-containing radical of formula (I) and octyltriethoxysilane, with the ratio octyltriethoxysilane:fluorine-containing oligomer = 50:1 - 1:50, with the following ratio of components (wt %): ultrafine diamond powder - 59.2-94.9; octyltriethoxysilane and fluorine-containing oligomer - 5.1-40.8. A method of producing said additive is also disclosed.

EFFECT: improved sedimentation resistance and tribotechnical properties of composite lubricant materials based on petroleum oil.

3 cl, 2 tbl, 6 ex

FIELD: chemistry.

SUBSTANCE: method includes the following steps: obtaining pores under the effect of electrolysis in a plate with thickness of 700-730 mcm and area of up to 32 cm2 of monocrystalline silicon, which is the anode, with p-type conductivity, doped with boron with concentration of about 10-19 cm-3, with resistivity of 3-7·10-3 ohm·cm, the surfaces of which are directed in parallel to the crystallographic planes in a glass-carbon cup, which is the cathode; wherein the electrolyte used is a solution consisting of different volume parts of hydrofluoric acid and ethyl alcohol; subsequent separation of the obtained porous layers from the remaining part of the bulk crystal by increasing the applied voltage by 5-90%, which changes the mechanism of the electrochemical process and switches from pore-formation to continuous polishing etching; washing the separated layers in ethanol and drying in air, followed by stepwise thermal annealing and grinding to fine powder state. Annealing is carried out first for 2 hours at 250°C and then for 20 minutes at 650°C in a hydrogen atmosphere.

EFFECT: obtaining silicon biocompatible nanocarriers with porosity of 40-80% and size of end-to-end pores of 5-20 nm.

3 cl, 7 dwg

FIELD: chemistry.

SUBSTANCE: invention relates to a method for synthesis of a fulleride of a metal nano-cluster and to a material containing a fulleride of a metal nano-cluster. The method for synthesis of a fulleride of a metal nano-cluster includes mechanically alloying metal nano-clusters with particle size between 5 nm and 60 nm with fullerene-type clusters by milling in a planetary mill, wherein fullerene molecules in the fulleride of a metal nano-cluster are preserved. The material obtained using said method contains a fulleride of a metal nano-cluster.

EFFECT: invention enables to obtain nano-blocks with improved mechanical properties, having high hardness, wherein metal nano-clusters are preserved during sintering.

12 cl, 2 dwg, 1 tbl, 2 ex

FIELD: nanotechnology.

SUBSTANCE: semiconductor film is influenced with continuous laser irradiation with quantum energy exceeding the band gap in the power range from 5 to 10 W, with a diameter of the laser beam on the film surface from 30 to 100 mcm, so that the intensity of exposure does not exceed 106 W/cm2, when scanning the surface of the film at a rate of 40 to 160 mcm/s.

EFFECT: invention simplifies the technical process, does not require any special equipment and enables to cover the devices with a characteristic period of location of the elements on the surface of 100 nm to 1 mcm.

8 dwg

FIELD: metallurgy.

SUBSTANCE: invention relates to metallurgy, and namely to cathode materials based on nanocrystalline Fe-Ni particles. A cathode for electrochemical hydrogen generation is made in the form of a steel substrate with a nanocomposite Fe-Ni coating applied onto its surface. The Fe-Ni coating with Ni content of 3-10 wt % has thickness of 0.5-0.9 mm and average grain size of up to 40 nm. A manufacturing method of a cathode for electrochemical hydrogen generation is characterised by the fact that a mechano-activated Fe-Ni powder nanocomposition with Ni content of 3-10 wt % is prepared and applied layer-by-layer onto the steel substrate, and layer-by-layer laser sintering is performed. Laser sintering is performed in vacuum with a fibre-optic pulse ytterbium laser at pulse generation frequency of 20000-100000 Hz and one-pulse time of 100 ns.

EFFECT: produced cathode is characterised by reduced hydrogen overvoltage.

2 cl, 2 tbl, 1 ex

FIELD: medicine.

SUBSTANCE: invention relates to method of obtaining nanocrystalline silicate-substituted carbonatehydroxyapatite (CHA), which includes mixing solutions of salts of calcium, phosphate and silicate, settling, filtering, washing from mother liquor and drying, solutions of calcium nitrate tetrahydrate, anhydrous disubstituted ammonium phosphate, sodium methasilicate pentahydrate are mixed with ratio of concentrations Ca/(P+Si) equal 1.70, and part of silicate-ions in the total quantity of sediment-forming anions (XSiO44=CSiO44/(CPO44+CSiO44)), constituting not more than 30 mol. %, pH is supported at level 9.00-12.00, obtained as a result of precipitation solid phase is kept under mother liquor for 2 days at temperature 22-25°C, then filtered out, washed with distillated water and dried at 80°C to constant weight. CHA considering created structure defectiveness of is characterised by higher solubility in physiological solution, and thereupon can be applied for formation of calcium-phosphate coatings on metal implants, as well as for creation of novel ceramic and composite materials for traumatology, orthopaedics and maxillofacial surgery.

EFFECT: increase of compound solubility.

1 tbl, 3 ex

FIELD: construction.

SUBSTANCE: peat and polymer mix for manufacturing of heat insulation products, including a base in the form of peat and a binding element, comprises four components, wt %: nanomodified peat of suspensions of polyvinyl acetate and cyclone dust of gas treatment of air of industrial enterprises (in terms of dry polyvinyl acetate in amount of 4-6% from mass of peat, dust - 2-3%) - 25%, portland cement CEM-II-32.5 - 40%, sodium carbonate - 10% and water - 25%.

EFFECT: increased fire resistance and strength characteristics at invariable density, reduced heat conductivity, increased homogeneity of structure and durability of heat insulation material.

1 tbl

FIELD: chemistry.

SUBSTANCE: invention relates to field of nanocapsulation in obtaining nanocapsules of glucoamine sulphate in xanthan gum envelope. In accordance with invention glucosamine sulphate is added by portions into suspension of xanthan gum in butyl alcohol, containing preparation E472c as surfactant. Mixture is mixed, after that hexane is added, obtained mixture of nanocapsules is filtered, washed with hexane and dried. Process is realised at 25°C for 15 minutes.

EFFECT: method in accordance with invention provides simplification and acceleration of process of obtaining glucosamine sulphate nanocapsules in xanthan gum and increase of output by weight.

2 ex

FIELD: chemistry.

SUBSTANCE: invention relates to field of analytic chemistry. Carrier is modified with organosilicon compound, containing -SH or -NH2 group, is processed with colloid gold solution. After that, sulphur-containing organic compound is covalently fixed on the surface of gold nanoparticles.

EFFECT: claimed is method of obtaining separation material, containing silicon dioxide-based carrier and gold nanoparticles.

3 cl, 6 dwg

FIELD: chemistry.

SUBSTANCE: method includes treating the surface of crystalline silicon by electrochemical etching in hydrofluoric acid solution with concentration of 20-30% while supplying current with surface density of 750-1000 mA/cm2 for 5-30 s to obtain hydrophobic silicon or supplying current with surface density of not more than 650 mA/cm2 for 5-30 s to obtain hydrophilic silicon.

EFFECT: method enables to obtain a surface with multimodal nano- or microporosity in a single step.

4 dwg

FIELD: chemistry.

SUBSTANCE: invention relates to cathode organic-inorganic hybrid material for secondary lithium-ionic sources of current with composition (C6H4N)*xV2O5*yH2O, where x=0.10-0.12, y=0.7-0.9 in form of nanorolls from 100 to 500 nm long and diameter from 10 to 20 nm with surface area 60 m2/g and pore diameter 20-30 nm. Invention also relates to versions of material obtaining.

EFFECT: claimed material possesses improved mechanical properties, high specific capacity and stability in time.

3 cl, 2 ex, 4 dwg

FIELD: chemistry.

SUBSTANCE: described is method of obtaining highly concentrated water nano-size polyurethane dispersion, which does not contain organic solvent, with concentration of basic substance 30-60%, which represents product of interaction of: A) at least, one polyisocyanate, which contains, at least, two isocyanate groups; B) one or several polyols with molecular weight (MW) from 1000 to 18000, which have, at least, two hydroxyl groups; C) one or several compounds, at least, with two OH-functional groups, which contain, at least, one carboxyl group, which can be transformed fully or partially into carboxyl group in presence of bases; D) possibly one or several polyols and/or glycidyl ethers of polyols with average molecular weight less than 500, which contain 2 or more hydroxyl and/or epoxy groups; E) one or several tertiary amines; F) one or several polyamines, which contain at least one NH2-group. Components (A), (B) and (C) are subjected to simultaneous interaction to degree of conversion of isocyanate groups 70-98%, if necessary component (D) is introduced into reaction mass, with the following complete or partial neutralisation of carboxyl groups of component (C) with component (E), dispersion in water, introduction of component (F); dispersion is heated and exposed at temperature from 20 to 90°C for from one to four hours. Also described are highly concentrated water nano-size polyurethane dispersion, obtained by claimed method, and its application for obtaining coatings for different substrates, especially flexible substrates, hermetics and glues.

EFFECT: obtaining coatings with high physical-mechanical properties, such as elasticity, durability, high water resistance.

18 cl, 14 ex, 15 tbl

FIELD: nanotechnology.

SUBSTANCE: albendazole is added in small portions to the suspension of sodium alginate in hexane in the presence of the preparation E472c while stirring, carbon tetrachloride is poured, the resulting suspension is filtered and dried at room temperature.

EFFECT: simplifying the process of production of the said capsules, the reduction of losses, and increase in the yield by weight.

2 ex

FIELD: chemistry.

SUBSTANCE: potassium losartan is added by portions into a suspension of xanthane gum in benzene in the presence of the preparation E472 with mixing at 1000 rev/sec. Then hexane is added. The obtained suspension of nanocapsules is filtered and dried at room temperature. The envelope:core ratio constitutes 3:1 or 1:5.

EFFECT: simplification and acceleration of the process of obtaining nanocapsules and an increase of output by weight.

2 ex

FIELD: chemistry.

SUBSTANCE: disclosed is a petroleum lubricating oil additive which contains ultrafine diamond powder treated with fluorine-containing oligomers with average molecular weight of 2200-4200 units and a general structural formula Rf-CF3 or Rf-COOH, where Rf is a fluorine-containing radical of formula (I) and octyltriethoxysilane, with the ratio octyltriethoxysilane:fluorine-containing oligomer = 50:1 - 1:50, with the following ratio of components (wt %): ultrafine diamond powder - 59.2-94.9; octyltriethoxysilane and fluorine-containing oligomer - 5.1-40.8. A method of producing said additive is also disclosed.

EFFECT: improved sedimentation resistance and tribotechnical properties of composite lubricant materials based on petroleum oil.

3 cl, 2 tbl, 6 ex

FIELD: medicine.

SUBSTANCE: to improve functional-aesthetic results of the plastic surgery of post-traumatic eyelid deformations, starting from 7-8 day from the moment of carrying out blepharoplasty of the post-traumatic cicatrical eyelid deformation, immediately after the removal of operational sutures, on the operated zone of the eyelid performed is a procedure of magnetic-photophoresis of a medical solution, made from 3000 ME of longidaze, 3 ml 25% solution of dimethylsulphoxide and 1 ml 0.25% solution of derinate, with 2 ml of the prepared solution being applied on an autodermal transplant and healthy skin surrounding it, with up to 1 cm indent from the wound edge, after 3-5 minutes a gauze pad, soaked with 2 ml of the remaining prepared solution, is applied on the processed surface of skin with the further 10 minute impact by a contact method by a travelling pulsed magnetic field of the apparatus "AMO-ATOS" with the frequency of pulse consecution of 5-10 Hz. Then, after the pad is removed, without the time interval, borders of the autodermal transplant and healthy skin closing are processed by infrared laser irradiation of a range of 0.89 mcm of the apparatus UZOR-2K with the frequency of the pulse repetition rate of 1500 Hz, in accordance with a contact labile method, 1 minute on each field, with carrying out 1 procedure daily for 10 days.

EFFECT: improvement of the quality of skin transplant engraftment, absence of secondary eyelid deformations, and reduction of the post-operational treatment duration.

1 tbl, 2 ex

Up!