RU2331424C2 - Topical formulation, containing spironolactone nanoparticles - Google Patents

Abstract

FIELD: medicine.

SUBSTANCE: invention relates to topical formulation with: spironolactone nanoparticles, containing medium-diameter nanoparticles measured by photonic correlation spectroscopy technique, in the range approximately 300 nm to 900 nm, and dispersion of solid polar lipids crystals in polar liquid, with additional stabiliser. The nanoparticles are included in crystal structure formed by solid polar lipids crystals; the lipids mentioned being directed their hydrophilic ends outside, and hydrophobic ends inside, relative to spironolactone nanoparticles. Items of the patent application are also as follows: method of producing the formulation, the crystal lattice arrangement of solid polar lipids crystals with spironolactone nanoparticles included, and use of the nanosuspension with structure mentioned for medication producing, intended for therapy in anti-androgen responding conditions.

EFFECT: achieving of high stability formulation and increased bioavailability of spironolactone.

17 cl, 4 ex, 8 tbl, 11 dwg

Description

The present invention relates to the use of spironolactone in the form of nanoparticles for topical treatment of a condition resulting from a reaction to antiandrogens. Such conditions include acne, hirsutism, androgenic alopecia, or rosacea.

Spironolactone is known as an aldosterone inhibitor and is used as a potassium-sparing diuretic. It goes on sale, in particular, under the name aldactone and can be used, for example, for the treatment of congestive heart failure. Spironolactone has an extremely low solubility in water, namely: 2.8 mg / 100 ml. Such low solubility can adversely affect the absorption of a drug substance by a living organism, which leads to poor bioavailability. Therefore, to achieve the desired level in the blood, increased doses of the drug substance are required. The low solubility of spironolactone also limits the preparation of drug substance formulations.

In other pharmaceutical applications, the antiandrogenic properties of spironolactone are used to treat various skin disorders, in particular, acne, hirsutism, androgenic alopecia and rosacea. Topical administration for the treatment of these disorders is the preferred method since it allows a significant reduction in systemic side effects. However, in this case, the poor solubility of the drug limits the development of effective and aesthetically acceptable formulations for topical application.

After oral administration, absorption of drugs from the intestine depends mainly on their solubility in intestinal fluids and their intestinal permeability. Poorly soluble preparations usually have low dissolution rates and create only a small concentration gradient in the intestinal mucosa, which can lead to low and unreliable absorption. Medicinal substances that have low solubility suffer from disadvantages with other methods of administration, for example, with topical.

Significant efforts are aimed at obtaining medicinal substances in the form of microparticles and nanoparticles. However, obtaining such fine particles is a non-trivial task and can create additional difficulties both in relation to the technical aspects of the process and in obtaining a satisfactory product. For example, it may be difficult to obtain particles with a stable and narrow range of sizes, especially in industrial production. In addition, it is necessary to obtain stable products, for example, nanosuspensions, however, microparticles and nanoparticles tend to form aggregates and flocculation, which has a negative effect on the stability of the product. A number of different approaches have been studied to obtain microparticles and nanoparticles.

US patent 5,091,188 describes a method for preparing solutions for injection of water-soluble preparations, which includes reducing the size of the crystalline drug substance in the range from 50 nm to 10 μm by ultrasonic treatment or other methods that cause intense shear in the presence of a phospholipid or other film-forming amphipathic lipid, the specified lipid covers the microcrystals of the drug.

US Pat. No. 5,145,684 describes particles of a crystalline drug substance containing a surface modifier without crosslinking and adsorbed on the surface, with an effective average particle size of less than about 400 nm. It is indicated that these particles are obtained by grinding in the presence of a grinding medium, for example, in a ball mill, a friction mill, a vibratory mill or a filled mill.

International patent application WO 96/14830 (US Pat. No. 5,858,410) describes a drug delivery composition that contains particles of a pure active compound, insoluble or slightly soluble in water. In this case, the average particle diameter is from 10 nm to 1000 nm, and the fraction of particles exceeding 5 μm in the total medium is less than 0.1%. Also described are methods of producing particles using or preferably without using a surface-active agent through cavitation (for example, using a piston homogenizer) or shear and shock forces (i.e., the principle of a jet stream).

Thus, there is a need for a topical composition containing spironolactone nanoparticles, which would eliminate these difficulties with obtaining drugs for local administration.

Applicants have shown that for topical application spironolactone in the form of nanoparticles can be successfully introduced into a cream base consisting of a crystal lattice of monoglycerides in water.

Thus, the first objective of the present invention is a topical composition of spironolactone containing nanoparticles with an average diameter measured by photon correlation spectroscopy in the range from about 300 nm to about 900 nm, preferably from 400 nm to 600 nm, introduced into the crystal lattice system, which contains a dispersion of solid crystals of polar lipids, while these lipids are turned outward with their hydrophilic side, and inward with their hydrophobic side towards spironolactone nanoparticles.

The composition is suitable for application to the skin in the treatment of dermatological conditions, which are known to be treatable with antiandrogens, in particular acne, hirsutism, androgenic alopecia and rosacea. The basis for a cream, consisting of a crystal lattice of monoglycerides, are described in patent applications WO 87/02582, WO 82/03173 and W093 / 20812. Examples of such monoglyceride crystal lattices are known as Crystalip ™.

Lipids can have a crystallization temperature of from 20 ° C to 100 ° C. Preferred lipid crystals are β crystals of a fatty acid monoglyceride having a chain length of 12-18 carbon atoms, or monoglycerol ethers having ester chains of the corresponding length, or ascorbic acid fatty acid esters with a fatty acid chain length of 12-18 carbon atoms or from their mixtures. Fatty acids and ethers may be saturated or unsaturated, preferably saturated.

Fatty acids may include lauric acid (C ₁₂ ), myristic acid (C ₁₄ ), palmitic acid (C ₁₆ ) or stearic acid (C ₁₈ ), however, C ₁₃ , C ₁₅ or C ₁₇ acids can also be used.

Preferred monoglycerides may be 1- or 2-monoglyceride, preferably 1-monolaurin, 1-monomyristin, 1-monopalmitin and 1-monostearin, or a mixture of two or more of them, in particular a mixture of 1-monolaurin and 1-monomyristin. Examples of unsaturated monoglycerides are monopalmitolein, monoolein, monolinolein and monoliniolenin.

The composition consists mainly of a dispersion of the above lipid crystals in water or another polar liquid, or mixtures thereof, contributing to the formation of crystals. Examples of polar liquids suitable for use in accordance with the invention are water, glycerin, propylene glycol and ethylene glycol or mixtures thereof, however, other suitable polar liquids can also be used.

Spirolactone is protected in the lattice up to the time of application when it is applied to the skin, while spirolactone comes into contact with the skin surface due to softening or melting of the crystalline structure of the shell.

In the general case, one could expect a noticeable increase in particle size after the introduction of very small solid particles into a matrix that contains hydrophilic as well as lipophilic structures. Oddly enough, this did not happen, and during the seven-month period a noticeable growth of the spironolactone crystal was not observed. In addition, a membrane model cream had a higher flow rate than a cream containing spironolactone in a form different from nanoparticles.

Pharmacists are well aware that particle sizes can be measured in various ways, which can cause differences in the described particle sizes. Such methods include photon correlation spectroscopy (PCS) and laser diffraction. In addition, the particle size can be expressed as the average particle size (for example, the average quantitative, average weight or average volumetric particle size). In this description, unless otherwise indicated, the average particle size is presented as the particle size. So, for example, D ₅₀ 500 nm indicates that 50% of the particle volume have a diameter of less than 500 nm. Alternatively, it can be indicated that particles having a diameter of less than 500 nm occupy 50% of the total volume occupied by the total number of particles.

If the particle size of the spironolactone according to the present invention is measured by laser diffraction, then D ₅₀ lies in the range of 350-750 nm, and D ₉₉ lies in the range of 500-900 nm.

Nanosuspensions and nanoparticles containing spirolactone according to the present invention preferably include a stabilizer to prevent aggregation of the nanoparticles. Such stabilizers, well known in the art, are described in more detail below.

In this description, nanoparticles containing spironolactone and nanosuspensions containing spironolactone according to the present invention are called spironolactone nanoparticles. It should be understood that this term also includes nanoparticles and nanosuspensions containing spironolactone together with a stabilizer.

Spironolactone nanoparticles according to the present invention can be obtained by any method known for producing nanoparticles, in particular, by high pressure homogenization.

Spironolactone nanoparticles can be obtained by cavitation of a coarse dispersion of spironolactone. Nanoparticles are preferably prepared using a high pressure piston homogenizer. Nanoparticles can be associated with a stabilizer. Such stabilizers, well known in the art, are described in more detail below.

To obtain nanoparticles, it is preferable to use the starting spironolactone material in the form of large particles, which preferably have a size of less than about 100 microns. If necessary, the particle size of spironolactone can be reduced to this level by conventional means, in particular, grinding. Large particles of spironolactone are preferably dispersed in a liquid medium containing a solvent in which the drug substance is practically insoluble. In the case of spironolactone, the liquid medium preferably contains an aqueous solvent and most preferably consists mainly of water. The concentration of spironolactone in the specified dispersion of large particles can be in the range from 0.1 to 50%. Then a coarse dispersion can be used to obtain nanoparticles in any known manner.

The preferred method is homogenization at high pressure, when the particle size is reduced mainly by cavitation. Most preferably, this is achieved using a high pressure piston homogenizer. In this method, a dispersion of large particles is passed at a high flow rate through a gap of about 25 microns wide. The static pressure exerted on the liquid drops below the internal vapor pressure of the liquid. Therefore, the liquid boils, forming gas bubbles in the gap zone. However, after the liquid exits the gap, normal pressure is restored and gas bubbles are destroyed. The powerful forces of the explosion directed inward, arising at the same time, reach a sufficient value to destroy large particles of the drug substance, and lead to the formation of nanoparticles.

Homogenization at high pressure can be performed at a pressure in the range from 100 to 3000 bar, preferably from 1000 to 2000 bar (from 10 ⁷ to 3 × 10 ⁸ Pa, preferably from 10 ⁸ to 2 × 10 ⁸ Pa) and at a temperature of the range from 0 to 50 ° C, preferably from 10 to 20 ° C, for example, about 15 ° C. Homogenization can be carried out cyclically to obtain the desired particle size or continuously for 2-30 hours, preferably for 2-10 hours.

The spironolactone nanosuspensions of the present invention preferably contain a stabilizer to prevent aggregation of the nanoparticles. The specified stabilizer can be entered at any convenient stage of the process of obtaining nanosuspension. Thus, for example, a surface-active agent can be added to the initial coarse dispersion before the formation of nanoparticles or after particle size reduction, for example, by high pressure homogenization. In an alternative embodiment, a stabilizer may be partially added before and after particle size reduction. It is preferable to introduce a stabilizer in a coarse dispersion. The concentration of the stabilizer in the coarse dispersion or nanodispersion can be from 0 to 10%.

The stabilizers that can be used to produce the nanosuspension according to the present invention can be selected from the group consisting of conventional stabilizers, as well as compounds described as surfactants and surface modifiers. Examples of suitable stabilizers include polyoxyethylene sorbitan fatty acid esters, for example, tween and span emulsifiers, polyoxyethylene stearates, polyoxyethylene alkyl esters, polyethylene glycols, block polymers and block copolymers, in particular poloxamers, for example, Lutrol F68, and poloxamines lecithins of various origins (e.g. egg lecithin and soya lecithin), chemically modified lecithins (e.g. hydrated lecithin), as well as phospholipids and sphingolipids, sterols (e.g. derivatives of cholesterol, as well as stigmasterol), esters of sugar and sugar alcohols with fatty acids or fatty alcohols (e.g. sucrose monostearate), ethoxylated mono- and diglycerides, ethoxylated lipids and lipoids, dicetyl phosphate, phosphatidyl glycerol, sodium cholate, , sodium taurocholate, sodium citrate, cellulose ethers and esters (e.g. methyl cellulose, hydroxyethyl cellulose, hydroxypropyl cellulose, sodium carboxymethyl cellulose), polyvinyl derivatives, in particular polyvinyl alcohol , polyvinylpyrrolidone, polyvinyl acetate, alginates, polyacrylates (e.g. carbopol), xanthan gum, pectins, gelatin, casein, gum arabic, cholesterol, tragacanth, stearic acid, calcium stearate, glyceryl sodium sulfonyl sulfonate sulfonate sulfate sulfonate sulfate sulfate chloride, alkyl-aryl polyester sulfonate, polyethylene glycols, colloidal silicon dioxide, magnesium aluminum silicate and phosphates.

A preferred stabilizer is docusate sodium, which is marketed in the form of a 70% propylene glycol solution under the brand name Octowet 70PG ™ (sodium dioctyl sulfosuccinate).

From the above description it is obvious that the process is carried out in a liquid medium and therefore the product of spironolactone nanoparticles is first obtained in the form of nanosuspension. If desired, the liquid medium can be removed, for example, by lyophilization or spray drying to obtain spironolactone nanoparticles in solid form. Moreover, if a stabilizer is present upon receipt of the nanosuspension, the corresponding dried product of the nanoparticles will contain the specified stabilizer.

After obtaining spironolactone nanoparticles, the composition according to the invention can be prepared as follows. The polar lipid is mixed with water and / or another polar liquid (in particular glycerol, ethylene glycol or propylene glycol) capable of forming structures with a crystal lattice of polar lipids. The resulting mixture has a concentration of water and / or polar liquid, respectively, 50-95% by weight. The mixture is heated to a temperature above the lipid phase transition temperature. The lipid phase transition temperature is defined as the minimum temperature at which a lipid particle in contact with excess water or polar liquid absorbs water or polar liquid, respectively, and turns into cylindrical or spherical crystalline structures having strong birefringence. Stirring, the mixture is maintained at a temperature exceeding the temperature of the phase transition of the lipid until the transition is complete. The mixture is then cooled with continuous stirring to ambient temperature or the desired temperature so that solid crystal lattices are formed. During this cooling, a characteristic crystalline structure is formed at a temperature of from about 30 to 35 ° C.

Spironolactone nanoparticles are dispersed in a mixture of polar lipid and water or polar liquid before or during the transition of the lipid into crystalline structures. To ensure the incorporation of spironolactone nanoparticles into the crystalline structure, they should be added before the mixture cools below 30-35 ° С.

If spironolactone nanoparticles are added after cooling the mixture below 30-35 ° C, a physical mixture is obtained, however, they do not form part of the crystalline structure. Therefore, the protection of recrystallization and its prevention, as well as increasing the particle size of the active component does not benefit spironolactone nanoparticles, since crystalline layers do not form around the active particles.

However, the nanosuspensions used according to the present invention to obtain a topical composition cannot withstand heat. When heated, agglomerates can form, and the active component can pass into solution at high temperatures. This can lead to recrystallization during cooling and cause a significant increase in particle size.

However, the applicants determined the composition, method and temperature of the introduction of nanosuspension, to ensure the formation of a crystalline structure after adding nanosuspension, while maintaining minimal thermal effect on nanosuspension.

The topical spironolactone nanoparticle formulations of the present invention preferably contain the active preparation in the form of a nanosuspension, most preferably in an aqueous solution. The pharmaceutical compositions of the present invention can be prepared by methods well known in the art.

Topical formulations according to the present invention can be prepared in the form of an ointment, cream, gel, liquid, aerosol or mousse. Aqueous formulations may directly contain nanosuspension, non-aqueous formulations may alternatively contain dried nanoparticles.

The second objective of the present invention is a topical composition containing spironolactone nanoparticles and intended for topical treatment of conditions that are known to be treated with antiandrogens, for example, acne, androgenic alopecia, hirsutism or rosacea.

A third object of the invention is the use of spironolactone nanosuspensions containing nanoparticles with an average diameter measured by photon correlation spectroscopy in the range from about 300 nm to about 900 nm, preferably from 400 nm to 600 nm, to obtain a medicament for treating a condition that is a reaction to antiandrogens, in particular, acne, hirsutism, androgenic alopecia, or rosacea. The medication can be adapted for local application. Nanoparticles can be incorporated into a cream base, which may consist of a crystal lattice of monoglycerides in water or other polar liquids.

This object of the invention extends to a method of treating a condition that is a reaction to antiandrogens, the method comprising administering a composition containing spironolactone nanoparticles to a patient in need of such treatment. The condition may be acne, hirsutism, androgenic alopecia, or rosacea.

A fourth object of the invention is compositions containing a crystal lattice system of solid crystals of polar lipids, wherein said lipids face outward with their hydrophilic side and inward toward the incorporated substance with their hydrophobic side, for use in topical treatment of acne. The crystal lattice system of solid crystals of polar lipids was previously described in patent application WO 87/02582 in relation to microcapsules.

A fifth objective of the invention is a method for producing a topical composition of spironolactone nanoparticles containing nanoparticles with an average diameter measured by photon correlation spectroscopy in the range of about 300 nm to about 900 nm, the method comprising introducing nanosuspension of spironolactone into an aqueous dispersion of polar polar solid crystals, and said lipids are turned outward with their hydrophilic side, and inward with their hydrophobic side towards spironolactone nanoparticles.

Nanosuspension can be introduced after cooling the mixture to 60 ° C-35 ° C, more preferably to 55 ° C-45 ° C, possibly to 50 ° C, before the mixture reaches the crystallization point. The temperature of the nanosuspension upon administration is preferably equal to room temperature, i.e. 20-25 ° C.

The preferred characteristics of the second and subsequent objectives of the invention are the same as for the first task with corresponding changes.

The following is a more detailed description of the invention with reference to non-limiting examples of implementation and the accompanying figures.

Figure 1 - image under the microscope of spironolactone nanoparticles according to the present invention immediately after their receipt. The distance between adjacent divisions of the scale corresponds to 0.01 mm.

Figure 2 - image under the microscope of spironolactone nanoparticles according to the present invention after their storage for 7 months at room temperature. The distance between adjacent divisions of the scale corresponds to 0.01 mm.

Figure 3 - image under the microscope of a commercially available spironolactone in a form different from nanoparticles. The distance between adjacent divisions of the scale corresponds to 0.01 mm.

Figure 4 is a typical calibration curve of spironolactone standards of 0.93-59.2 μg / ml.

Figure 5 is a graph of the average flow of spironolactone from nanosuspension (2% by weight) and water cream (2% by weight) (n = 4, average +/- standard error)

y = 35.552x-20.258r2 = 0.991 Spironolactone Aqueous Cream 2%

y = 42.097x-26.784r2 = 0.989 Spironolactone nanosuspension 2%

Figure 6 - the average diameter of the zone S. epidermidis spironolactone composition Crystalip ^™ compared with 2% by weight of spironolactone in water cream (British Pharmacopeia - British Pharmacopoeia) (mean ± standard deviation, n = 5).

Figure 7 - the average diameter of the zone P. acnes spironolactone composition Crystalip ^™ compared with 2% by weight of spironolactone in water cream (mean ± SD), n = 5).

или 70°С

с последующим охлаждением по сравнению с наносуспензией, не подвергавшейся нагреванию

.Figure 8 - particle size distribution of spironolactone nanosuspension after heating to 50 ° C

or 70 ° C

followed by cooling compared to nanosuspension, not subjected to heating

.

или 70°С

с последующим охлаждением по сравнению с наносуспензией, не подвергавшейся нагреванию

.Figure 9 - particle size distribution of spironolactone nanosuspension 24 hours after heating to 50 ° C

or 70 ° C

followed by cooling compared to nanosuspension, not subjected to heating

.

Figure 10 - viscosity of the mixture after administration of the OctowetTM composition at 50 ° C, 70 ° C and room temperature.

Figure 11 - dependence of the viscosity of the mixture on its composition.

EXAMPLES

Example 1: Obtaining a topical composition containing spironolactone nanoparticles

Obtaining spironolactone nanoparticles

Table 1 shows examples of the compositions of spironolactone nanoparticles intended for introduction into the crystal structure according to the present invention. Spironolactone nanoparticles can be obtained as follows. The preparation of an aqueous stabilizer solution was poured into water or an injection buffer while stirring with a magnetic stirrer until a clear solution was obtained. A suspension was obtained by wetting spironolactone with an appropriate amount of an aqueous solution of a surfactant. The resulting suspension was dispersed using a high performance dispersant. Suspensions were kept under stirring with a magnetic stirrer to prevent foaming.

The resulting suspensions were passed through a high pressure piston homogenizer to obtain a nanosuspension. Compounds 1-7 were prepared using Avestin C5 ^™ , and formulations 8 and 9 were prepared using Avestin C50 ^™ . During homogenization, the particles of the preparation were destroyed under the action of cavitation and shear forces, and micro- and nanoparticles were formed. Particle sizes were determined by photon correlation spectroscopy (PCS) using a Zetasizer 3000 HS ™ (Malvern). D ₅₀ and D _{90 were} measured by laser diffraction using a Coulter LS230 instrument.

Table 1 Structure one 2 3 four 5 6 7 8 9 Spironolactone,% 10 10 twenty 10 10 10 10 10 10 Sodium lauryl sulfate,% one 0.1 0.4 0.1 Lutrol F68% - one one 0.4 0.1 0.4 - - - NaCl - - - - - 0.9 - - - Octowet 70 (docusate sodium),% - - - - - - 0.5 (0.35) 0.5 (0.35) 0.5 (0.35) Water up to 100% Process modes Pressure (bars) 1500 1500 1500 1500 1500 1500 1250 1500 1500 Time (min) 90 180 180 180 180 300 300 1080 Sample Volume (ml) 40 40 40 40 40 40 40 one hundred 500 results D ₅₀ (μm) 1,69 0.85 1.06 0.84 0.88 0.86 0.78 0.54 0.539 D ₉₀ (μm) 4.39 1.83 2.49 1.92 1.82 1,5 1.8 0.68 0.772 PCS average diameter - 581 880 608 681 656 609 415 436 PI - 0.7 0.2 0.15 0,03 0.1 0.2 0.05 0.1 Obtaining the composition of Crystalip ^™

Table 2 presents examples of preparations containing spironolactone nanoparticles in the form of a topical cream obtained from formulations 7, 8 and 9 shown in table 1. Topical nanosuspension preparations were prepared as follows.

Heated water to 70 ° C and added propylene glycol. Monoglycerides were melted at 70 ° C and molten monoglycerides were added to the aqueous phase with stirring at a speed of 70 rpm. Then began the cooling of the mixture. When the temperature of the mixture reached about 50 ° C., the stirring speed was increased to 95 rpm and cold nanosuspension was added. At 35 ° C, when a β-crystalline structure began to form, the stirring speed was reduced to 75 rpm.

table 2 Structure BUT AT FROM D Spironolactone nanosuspension (formulations 7, 8 or 9 of table 1) twenty 10 twenty twenty Glycerol monolaurate 7 6 6 5 Glycerin monomyristate 21 eighteen eighteen fifteen Propylene glycol - 10 10 twenty Water 52 56 46 40

To determine the optimal compositions and method for producing a topical composition containing spironolactone nanoparticles in accordance with the present invention, the following experiments were carried out.

Choosing the Optimum Crystalip ^™ Composition

Two batches of Crystalip ^™ formulation were prepared as indicated in Table 3.

Table 3 Placebo Crystalip ^™ Composition Batch number Glycerol monolaurate Monomyristate glycerin Propylene glycol (PG) Water 4011-001 / 01 7% 21% / 72% 4011-001 / 02 5% fifteen% twenty% 60%

For the first batch, a characteristic exothermic peak was observed upon cooling. At the end of the cooling stage, the cream was very viscous, however, did not have the gloss typical of the β-crystalline structure. During storage, this shine began to appear.

For the second batch, an exothermic peak was not observed, however, the material was glossy. Viscosity was apparently lower than that of the first batch.

Then tested the composition with a lower content of propylene glycol (PG) (table 4), because there were concerns about the possible occurrence of irritation due to high concentrations of PG. Propylene glycol is advisable to keep in the composition, because it can increase the antibacterial effectiveness of the base and, therefore, enhance the penetration of active components into the skin. The following batches of material were produced:

Table 4 Crystalip ^™ parties Batch number Glycerol monolaurate Monomyristate glycerin Propylene glycol (PG) Water 4011-001 / 03 5% fifteen% 10% 70% 4011-001 / 04 6% eighteen% 10% 66%

For a batch with 20% monoglyceride (MG), the viscosity decreased at about 40 ° C, and the mixture again became liquid immediately after the viscosity began to increase.

Therefore, the MG content was increased to 24%. The viscosity also decreased at about 40 ° C, however, it again increased with subsequent cooling. An exothermic peak was observed at 33 ° C, and the finished cream was glossy, which indicated the formation of ultimately a β-crystalline structure.

Surfactant and Crystalip ^™ Compatibility Tests

Crystalip ^{™ was} previously found to be compatible with Myrj 59 and Span 20, which are non-ionic surfactants. However, Octowet 70PG, which is an anionic surfactant, is used for spironolactone nanosuspension. Octowet 70PG is also known as sodium dioctyl sulfosuccinate and is a 70% solution (70% DOSS) in water and propylene glycol.

To obtain a cream containing 2% spironolactone, it is necessary to introduce 20% spironolactone nanosuspension, since the nanosuspension contains 10% active spironolactone and 0.5% surfactant. Therefore, when testing the compatibility of Crystalip ^™ with Octowet, a 20% Octowet solution was used and part of the aqueous phase was replaced.

The compatibility of the Octowet 70 PG solution with the selected Crystalip ^™ formulation (10% PG + 24% MG) was tested. Three batches of Crystalip ^™ were prepared (Table 5), to which a 20% Octowet 70 PG solution was added at various temperatures: 70 ° C, 50 ° C and at room temperature.

Table 5 Crystalip ^™ batches containing Octowet solution Batch number Glycerol monolaurate Glycerol monomyristate Propylene glycol Octowet Solution
(0.5%) Water T ° C * 4011-001 / 05 6% eighteen% 10% twenty% 46% 70 4011-001 / 06 6% eighteen% 10% twenty% 46% fifty 4011-001 / 07 6% eighteen% 10% twenty% 46% room pace. * T ° C: temperature of adding a solution of surfactant

All three parties formed an exothermic peak and had a brilliance. Lot number 05 and 06 had an acceptable viscosity, while lot number 07 after adding Octowet was insufficient.

Based on the foregoing, it was concluded that the selected Crystalip ^™ formulation is compatible with a 20% 0.5% Octowet solution. Octowet solution is better to add at a temperature higher than the crystallization point than at room temperature.

Adding spironolactone nanosuspension to Crystalip ^™

The nanosuspension contained 10% spironolactone and 0.5% Octowet 70PG.

The particle size of fresh nanosuspension, determined by the method of laser diffraction (COULTER), was:

D50 = 0.686

D90 = 1,033

D99 = 1,033

Particle size determined by photon correlation spectroscopy (Zetasizer 3000HS): Z average = 476 nm.

The compatibility tests described above showed that adding materials at room temperature is not the best option. Therefore, it turned out to be necessary to study the heating of nanosuspension. The particle size was measured before and after heating the suspension.

Conducted the following experiment.

Two nanosuspension samples were heated to 50 ° and 70 ° C, respectively. It was held at the indicated temperature for 10 minutes, and then the samples were cooled to room temperature. Particle size was measured by photon correlation spectroscopy and laser diffraction at the following time points:

- after ultrasonic treatment, before heating (base sample),

- after cooling (t0),

- after 24 hours (t24h).

The results presented in table 6 and 7 are also shown in figures 8 and 9.

Table 6 Particle size determination by laser diffraction (Coulter) Distribution
particle sizes Sample Base 50 ° C - t0 70 ° C - t0 50 ° C - t24h 70 ° C - t24h D50 0.579 nm 0.638 nm 0.700 nm 0.634 nm 0.694 nm D90 0.740 nm 0.878 nm 1,053 nm 0.861 nm 1.572 nm

Table 7 Particle size determination by photon correlation spectroscopy (PCS) (Zetasizer 3000HS) Sample Base 50 ° C - t0 70 ° C - t0 50 ° C - t24h 70 ° C - t24h Particle size (nm) 456.3 469.7 538.3 Not
defined * Not determined* (*): the laser diffraction measurement showed too large, so PCS measurement was not carried out

The results obtained by heating nanosuspension show a sharp increase in particle size at 70 ° C, especially after exposure for the next 24 hours. This suggests that the drug partially passes into solution, and then recrystallizes. The particle size also increased at 50 ° C; however, apparently, no such strong recrystallization occurred during the next 24 hours.

Based on the results, it was concluded that a very short shutter speed at 50 ° C is acceptable. Therefore, they decided to make the final batch as follows: to prepare Crystalip ^™ with a reduced content of the aqueous phase. Subject nanosuspension to ultrasonic treatment, but do not heat. After reaching Crystalip ^™ temperature of 50 ° C add a cold suspension, which will lead to a rapid decrease in temperature and minimizes the effects of heat on the spironolactone suspension. Crystalip ^™ crystallization, which occurs at lower temperatures, will also not interfere with this process.

A batch of material was obtained according to the above procedure, with cold nanosuspension added to Crystalip ^™ at 50 ° C (table 8).

Table 8 Crystalip ™ batch containing spironolactone nanosuspension Batch number Glycerin mono
laurat Glycerol monomyristate Propylene glycol Nanosuspension Water 4011-001 / 08as 6% eighteen% 10% twenty% 46%

This party was successful. The final cream obtained had good viscosity, gloss and a uniform composition.

Viscosity measurement (see table 9).

Table 9 Sample Structure Bioglan 20% MG-20% PG 4011-001 / 05rs 24% MG-10% PG-20% Octowet solution (added at 70 ° C) 4011-001 / Obrs 24% MG-10% PG-20% Octowet solution (added at 50 ° C) 4011-001 / 07rs 24% MG-10% PG-20% Octowet solution (added at Sample Structure room temperature) 4011-001 / 08as 24% MG-10% PG-20% nanosuspension 4011-001 / 13rs 28% MG 4011-001 / 17rs 20% MG-20% PG

The results of measuring the viscosity of the compositions presented in the above table 9 are also shown in figures 10 and 11.

Tests of the second batch of nanosuspension: 3011-05anl

Since the nanosuspension was not fresh enough after the tests described above, we decided to make a new suspension of spironolactone and introduce it into Crystalip ^™ only a few days after isotation. The nanosuspension used contained 10% spironolactone and 0.5% Octowet 70PG. We also investigated the possibility of introducing into Crystalip ^™ 30% nanosuspension instead of 10%.

The particle size of fresh nanosuspension, determined by the method of laser diffraction (COULTER), was:

D50 = 0.443

D90 = 0.657

D99 = 0.738

Particle size determined by photon correlation spectroscopy (Zetasizer 3000HS): Z average ≈ 419.3 nm.

Two new batches of Crystalip ^™ were prepared - 4014-000 / 01ac and 4014-000 / 02ac, containing 20% and 30% nanosuspension, respectively. The method was identical to the method of manufacturing a batch 4011-001 / 08as (the introduction of nanosuspension at 50 ° C in the cooling step). Nanosuspension was prepared on the eve of Crystalip ^™ manufacture. In both of these batches, the pH value was adjusted corresponding to a commercially available cream (Spiroderm 5% with a pH of 4.16). The composition of both parties is presented in table 10.

Table 10 Batch number Glycerol
monolaurate Glycerol
monomyristate Propylene glycol 3011-05an1 Water Lemon acid Sodium hydroxide 4014-000 / 01ac 6% eighteen% 10% twenty% 45.5% 0.5% Up to pH 4.16 4014-000 / 02as 6% eighteen% 10% thirty% 35.5% 0.5% Up to pH 4.16

Both creams had a shine, however, the second - 4014-000 / 02as, apparently, had a higher viscosity. With the introduction of cold nanosuspension, the viscosity increased faster in the case of a batch containing 30% nanosuspension. In addition, a batch containing 30% nanosuspension seemed less uniform due to the increase in viscosity.

The pH and density of these batches, measured the day after manufacture, are shown in table 11.

Table 11 Batch number pH Density
(g / cm ³ ) 4014-000 / 01ac 4.29 0.989 4014-000 / 02as 4.22 0.984

Example 2: Spironolactone Particle Size Before and After Storage

In figures 1, 2 and 3 presents micrographs of spironolactone according to the invention immediately after receipt, after storage for 7 months and their comparison with commercially available spironolactone. The figures also show a scale with a division value of 0.01 mm or 10 μm.

The particles shown in figures 1 and 2 are so small that they are almost invisible using an optical microscope. After storage for 7 months, an increase in particle size is not observed. In contrast, commercially available Spiroderm spironolactone (Figure 3) contains crystals up to 20 microns in size.

Example 3: Study of flow parameters

Using a Franz cell, the spironolactone flux (2%) flowing through artificial membranes was measured from the nanosuspension introduced into the Crystalip ^™ matrix, and it was compared with 2% by weight spironolactone in B.P. water cream (see table 12).

Table 12 Material Provider Crystalip ^™ Spironolactone Nanususpension 2% SkyePharma, Switzerland Party No. 4014-000 / 06atc Spironolactone Party №510 / 0 Water cream V.R. Hillcross, UK Party No. 28076 Ethanol brand AnalaR VWR International Ltd., UK Sodium dihydrogen phosphate batch No. L298 Merckeurolab Deionized water Elga Ltd., United Kingdom HPLC Purity Acetonitrile (High Resolution Liquid Chromatography) Rathburns Chemicals Ltd., United Kingdom Regenerated cellulose membrane NBS-Biological, UK

Methods for studying flow

Crystalip ^{™ was} obtained from SkyePharma AG. During the experiments, a commercially available spironolactone formulation was not available for comparison. Therefore, the composition for comparison was prepared in a standard cream matrix as follows. Briefly, 100 mg of spironolactone powder was weighed accurately and mixed with 4.90 g of B.P. water cream to obtain a water cream preparation containing 2% by weight of spironolactone in a form different from the nanoparticles.

The study of diffusion in artificial conditions

Ethanol-phosphate buffer (20:80 by volume, pH 4.5) was used as a liquid in the receiver to maintain the stability of spironolactone and immersion conditions. A regenerated cellulose membrane was used as an artificial membrane.

The study of diffusion using Franz cells

To conduct diffusion experiments, individually calibrated Franz diffusion cells were used with an average diffusion surface area of 0.56 ± 0.03 cm ² and an average receiver volume of 1.83 ± 0.02 ml. Spectra / Por® cellulose membranes were cut to size and immersed for 30 minutes in deionized water to remove the preservative (0.1% sodium azide), wiped with a cloth to remove liquid from the surface, and placed on Franz cells. The receiver liquid was poured into a Franz cell, constantly stirring with a magnetic stirrer, and kept at 32 ° C. The membranes were aged to create equilibrium with the receiver phase for 30 minutes before applying the formulations. Each formulation (200 μl) was applied to the surface of the membrane using a Finnpippette® direct displacement device. Five time points (1, 2, 4, 6, and 8 hours) were examined, taking 200 μl of receiver fluid from the knee of a Franz cell. Each sample taken was replaced with an equal volume of fresh, preheated (32 ° C) receiver fluid. As the experiment was conducted, the losses of the receiver fluid in the Franz cell due to evaporation were compensated to maintain a constant volume. Samples were analyzed using HPLC (high performance liquid chromatography) using the following chromatographic modes:

Speaker: Hypersil 3 μm phenyl BDS (serial number 182862) Column Length: 150 × 4.60 mm Column Temperature: 30 ° C Mobile phase: 50 mM phosphate buffer: acetonitrile (70:30 v / v) Flow rate: 1.0 ml / min UV wavelength: 238 nm Injection volume: 10 μl Travel time: 15 minutes

Obtaining standard curves for spironolactone

Spironolactone standards were prepared in the receiver fluid and calibration curves were constructed in the range of 0.93-59.2 μg / ml. The calibration curves with r2> 0.999 were considered acceptable. A typical curve is shown in figure 4.

Data analysis

The spironolactone content in the receiver fluid was adjusted for sampling. We plotted the dependence of the cumulative mass of spironolactone penetrating through a unit surface of the membrane on the square root of time, while the slope of the linear portion of the graph was considered the stationary state of the flow. For statistical determination of the significance of the difference in the separation of spironolactone from 2% spironolactone nanosuspensions and 2% spironolactone aqueous cream, a test was used according to the Student criterion.

results

Figure 5 shows a graphical representation of the average cumulative mass of spironolactone penetrating through a surface unit (μg / cm ² ) for two formulations containing spironolactone. These profiles indicate the steady state flow of both compositions. The rate of release of spironolactone from spironolactone nanosuspensions (2% by weight) was significantly higher than from aqueous cream (p = 0.08).

Example 4: Determination of the zone of inhibition

The antibacterial effect of two spironolactone formulations, namely, spironolactone nanosuspension with 2% by mass and coarse-grained spironolactone coarse suspension with 2% by mass, and their respective placebo were compared. As a comparative composition used water cream B.R. (own production). Used the following materials:

Material Provider Staphylococcus epidermidis Oxoid Ltd, UK (ATCC 12228) Pseudomonas acne Central Public Health Laboratory, UK (NTC 737) S&S Disks for antibiotic analysis (filter paper), 1/4 inch diameter Aldrich Chemical company, CLUA Water cream V.R. Hillcross, UK Party No. 28076 Spironolactone SkyePharma, Switzerland Party №510 / 0

Methodology

The spironolactone formulations Crystalip ^™ and placebo were prepared according to Table 13. Since there was no commercially available spironolactone formulation for comparison, a formulation was prepared for comparison in a standard cream matrix as follows. Briefly, 100 mg of spironolactone powder was weighed accurately and mixed with 4.90 g of B.P. water cream to obtain an aqueous cream containing 2% by weight of spironolactone in a form other than nanoparticles.

A batch of 500 g was manufactured in a laboratory reactor IKA-LR1000.2.

Structure 4014-000 / 06atc Crystalip ™ nanosuspension 2% MG (monoglycerides) 24 PG (propylene glycol) 10 Nanosuspension - 10% spironolactone twenty Spironolactone coarse / Octowet 0.5% by weight / Lemon acid 0.1 Sodium hydroxide to pH 4.16 PPI water 45.9 Physical properties pH 4.31 Density (g / cm ³ ) 0,900 Viscosity (cP) 20 rpm 42'327 50 rpm 23'780

20 ± 1 mg of each of the following samples was carefully transferred onto the surface of 1/4 inch diameter discs for antibiotic analysis.

1. SkyePharma AG, Crystalip ^™ , spironolactone nanosuspension, 2% by mass, 4014-000 / 06atc.

2.2% by weight spironolactone in BP water cream.

Using a pair of sterile tweezers, antibiotic discs coated with each of these formulations were placed on the surface of agar plates with seeded organisms.

Plates of S. epidermidis were incubated for 24 hours at 37 ° C.

P. acne plates were incubated in anaerobic vessels for 72 hours.

Inhibition zones were measured using a pair of compasses.

results

Figures 6 and 7 show the average inhibition zones for both formulations, determined by plating with S. epidermidis and P. acne, respectively. A spironolactone nanosuspension Crystalip ^™ formulation has a significant effect on these acne-related bacteria. The water cream VR containing 2% by weight of spironolactone (comparative composition) did not show zones of inhibition.

Thus, the matrix of the base gives the composition antibacterial properties (when exposed to S. epidermidis and P. acne), which are not associated with spironolactone. The composition does not contain additional antibiotics or preservatives. A comparative product that did not affect these acne-related microorganisms is canned with phenoxyethanol. Therefore, the product proposed by the applicants can have a therapeutic effect on acne both through the hormonal activity of the drug and due to the antibacterial effectiveness of the matrix in which it is contained.

Claims (17)

1. A topical composition for treating conditions that can be treated with antiandrogens with spironolactone nanoparticles, containing medium-diameter nanoparticles measured by photon correlation spectroscopy in the range of about 300 nm to about 900 nm, and a dispersion of polar polar solid crystals in polar liquid, moreover, spironolactone nanoparticles are included in the crystalline structure formed by solid crystals of polar lipids, while these lipids are reversed by their hydrophilic side Rouge, a hydrophobic side - inwards towards the spironolactone nanoparticles, and optional stabilizer. 2. The composition according to claim 1, characterized in that it contains nanoparticles of average diameter, measured by photon correlation spectroscopy, in the range from about 400 nm to about 600 nm. 3. The composition according to claim 1, characterized in that the crystallization temperature of the lipid is from 20 to 100 ° C. 4. The composition according to claim 1, characterized in that the lipid crystals are β-crystals of monoglyceride fatty acids containing 12-18 carbon atoms, or esters of fatty acids of ascorbic, phosphoric or lactic acids or simple monoglycerol esters or mixtures thereof. 5. The composition according to claim 4, characterized in that the monoglyceride is 1-monolaurin, 1-monomyristin, 1-monopalmitin or 1-monostearin or a mixture of two or more of these compounds. 6. The composition according to claim 1, characterized in that the polar liquid is selected from the group comprising water, glycerin, ethylene glycol, propylene glycol and mixtures thereof. 7. The composition according to claim 1, characterized in that it is intended for topical treatment of acne, hirsutism, androgenic alopecia or rosacea. 8. The composition according to claim 1, characterized in that the composition contains the active drug in the form of nanosuspension. 9. The composition of claim 8, wherein the nanosuspension is an aqueous nanosuspension. 10. The composition according to claim 9, characterized in that the stabilizer is docusate sodium. 11. The use of spironolactone nanosuspension containing spironolactone nanoparticles having an average diameter measured by photon correlation spectroscopy ranging from about 300 nm to about 900 nm, enclosed in a crystalline structure formed by solid crystals of polar lipids, with these lipids facing their hydrophilic side outward, and the hydrophobic side - inward towards the spironolactone nanoparticles, in a polar liquid, and additionally containing a stabilizer to obtain honey a drug intended to treat a condition treatable with antiandrogens. 12. The use according to claim 11, characterized in that said condition is selected from the group consisting of acne, hirsutism, androgenic alopecia, or rosacea. 13. The use according to claim 11, characterized in that the medication is adapted for topical application. 14. The use according to claim 11, characterized in that the nanoparticles are introduced into the base for the cream. 15. The application of clause 14, wherein the cream base consists of a crystal lattice of monoglycerides in water or other polar liquids. 16. A crystal lattice system that contains a dispersion of solid crystals of polar lipids in a polar liquid and spironolactone nanoparticles of average diameter, measured by photon correlation spectroscopy, in the range from about 300 nm to about 900 nm, with these lipids facing out with their hydrophilic side, and hydrophobic side - inward towards the spironolactone nanoparticles included in the lattice, intended for topical treatment of acne. 17. A method for producing a topical composition with spironolactone nanoparticles containing nanoparticles having an average diameter measured by photon correlation spectroscopy in the range of about 300 nm to about 900 nm, said method comprising introducing nanosuspension of spironolactone into an aqueous dispersion of polar polar solid crystals by temperature exceeding the lipid phase transition temperature, followed by cooling, and spironolactone nanoparticles are dispersed in a mixture of polar lipid and water or and polar fluid before or during the transition of the lipid into crystalline structures, while these lipids are turned outward with their hydrophilic side, and inward with the hydrophobic side towards spironolactone nanoparticles.

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