NL8802008A - PHARMACEUTICAL PREPARATIONS FOR THE TOPIC APPLICATION, AND METHOD FOR PREPARING THE SAME - Google Patents

Description

NO 35 312 1 S--

Pharmaceutical preparations for the topical application, and method for their preparation »____

The invention relates to new antimicrobially active pharmaceutical preparations for the topical application. More specifically, the invention relates to antimicrobial pharmaceutical preparations for the topical application on the basis that, as an antimicrobially active substance, a compound from the group of active substances, which in its molecule contains an imidazole substituted with a lower alkyl group, in combination with one or more optionally substituted benzoic acids is used, the combination resulting in a synergistic antimicrobial activity.

EP-A-0.007.595 discloses pharmaceutical preparations containing the anti-mycotic active ingredient clotrimazole (1 - [(2-chlorophenyl) diphenylmethyl] -IH-imidazole) for external use, in particular one containing this active ingredient new gel, cream or also liquid preparations containing crotamitone (N-crotonyl-N-ethyl-o-tolufdine) as a solvent.

Furthermore, EP-A-0 070 525 discloses pharmaceutical preparations containing antimycotic active substances containing an imdazole ring in the molecule, such as e.g. miconazole, econazole or also isoconazole, which are dissolved in crotamitone, peppermint oil or also methyl salicylate, the latter also having pharmaceutical effects as skin preparations, however none of them are anti-mycotic.

As a result of further investigations, as also described in more detail later, it was surprisingly and unexpectedly found that the combination consisting of an antimicrobial active substance from the group of active substances, which in its molecule is an imidazole substituted with a lower alkyl group according to a compound of the formula 1, in which at least one of the substituents R 1, R 2 and R 3 represent a halogen atom, and the other hydrogen also both contain its pharmaceutically acceptable acid addition salts or according to a compound of the formula 2, wherein R4 represents a halogen atom, with one or more optionally substituted benzoic acids and optionally conventional pharmaceutical carriers and / or excipients in pharmaceutical preparations for the topical application, has an extraordinary synergistic antimicrobial activity.

Optionally substituted benzoic acids are those which contain one or more halogen atoms, hydroxyl or also lower alkoxy groups as substituents at various locations on the benzene ring. Preferred are <8802008 2 "optionally monosubstituted benzoic acids which carry the substituents in the ortho position relative to the carboxyl group in the benzene ring.

Of particular interest in the above combinations is the use of o-hydroxybenzoic acid, salicylic acid and / or the unsubstituted benzoic acid.

Halogen is bromine, iodine or fluorine, but in particular chlorine.

Lower alkoxy is ethoxy, n-propoxy, isopropoxy, n-butoxy, iso-butoxy, sec-butoxy or tert-butoxy, especially methoxy.

The combination of the above active substances has an unexpected synergistic antimicrobial effect of the individual components.

The invention particularly relates to antimicrobial pharmaceutical preparations for the topical application, consisting of the combination of a compound of the formula 1, in which at least one of the substituents R1, R8 and R3 is a chlorine atom and the remaining hydrogen as well as their pharmaceutically acceptable acid addition salts, such as e.g. miconazole (Ri = Rg = Cl, R3 = H), econazole (R2 = Cl, R] _ = R3 = H) and isoconazole (Ri = R3 = Cl, R2 - H) or a compound of the formula 2, wherein R4 represents a chlorine atom, such as e.g. clotrimazole and one or more optionally substituted benzoic acids and optionally conventional pharmaceutical carriers and / or excipients.

The invention relates in particular to those topically applicable pharmaceutical preparations which, because of the synergistic effect, have particularly effective antimicrobial properties, consisting of the combination of miconazole as a compound of the formula 1, wherein R1 and R2 are a chlorine atom and R3 is hydrogen and a pharmaceutically acceptable acid addition salt thereof or from a compound of formula 2, wherein R 4 represents a chlorine atom, clotrimazole and salicylic acid and / or benzoic acid and optionally conventional pharmaceutical carriers and / or optional auxiliaries.

Preferably, the invention relates to those particularly effective topically applicable pharmaceutical preparations, which contain a combination of miconazole nitrate or clotrimazole and salicylic acid and / or benzoic acid and optionally conventional pharmaceutical carriers and / or excipients.

All physiologically compatible acids are suitable for the preparation of the pharmaceutically acceptable acid addition salts of the compounds of the formula I. To this end, the following acids will be mentioned as examples: 8802008: hydrogen halides, such as e.g. hydrogen chloride and hydrogen bromide, especially hydrogen chloride, further sulfuric acid, phosphoric acid, nitric acid, perchloric acid, aliphatic, alicyclic, aromatic or heterocyclic carboxylic acids or sulfonic acids, such as e.g. formic, acetic, propionic, succinic, glycolic, lactic, malic, tartaric, citric, ascorbic, maleic, hydroxymalephic, pyruvic, phenylacetic, benzoic, p-aminobenzoic, anthranilic, p-hydroxybenzoic, p-hydroxybenzoic, aminosalicylic acid, embonic acid, methane, ethanesulfonic acid, hydroxyethanesulfonic acid, ethylene sulfonic acid, halobenzenesulfonic acid, toluenesulfonic acid, 1,5-naphthalenedisulfonic acid or sulfanilic acid, methionine, tryptophan, lysine or also arginine.

The pharmaceutical preparations according to the present invention have antimicrobial, in particular antimycotic and antibacterial, properties. For example, its e.g. effective against those microorganisms which occur with skin lesions and lesions, such as e.g. Trichophyton species, for example Trichophyton mentagrophytes, Epider-maphyton, for example Epidermaphyton floccosum, Microsporon, for example Microsporon felineum, Aspergillus species, e.g. Aspergillus niger and Aspergillus funigatus, yeast fungi, such as Pytyrosporum species, Candida species, for example, Candida albicans and gram positive bacteria, e.g. Staphylococcus species, for example Staphylococcus aureus, Pseudomonas aeruginosa, Propioni bacterium and Coryne bacterium acnes. The list of these microorganisms does not in any way represent a limitation of the controllable germs, but is merely illustrative.

The pharmaceutical preparations which can be prepared according to the invention can be used as antimycotic active agents for the treatment of skin infections which are determined by dermatophytes and sprout-shaped fungi, such as e.g. dermatomykoses and system mycoses or also as an antibacterial active agent, for the treatment of acne, dandruff and other skin conditions. Furthermore, the combinations which can be prepared according to the invention also have anti-pruritic properties and can be used for the treatment of pruritus diseases.

The pharmaceutical preparations according to the invention for the topical application contain, in addition to the combinations of a compound of the formula I, respectively. 2 and one or more optionally substituted benzoic acids also pharmaceutically acceptable carriers or excipients. The daily dose of 40 active ingredients depends on age or. 88 0 2008.

4 »- other individual circumstances of the patient and also according to the nature of the formulation.

Preferred concentrations of the combined active substances in the formulated preparations are the range from 0.01-10% (weight) of the lower alkyl substituted imidazoles of the formula 1 or 2, preferably 0 1-2% (weight) and 0.5-25% (weight) for the one or more optionally substituted benzoic acids, but preferably 1-15% (weight).

Suitable pharmaceutical preparations for topical application are primarily creams, ointments and gels, furthermore powders, pastes, lotions, tinctures, foams, sprays, aerosols, solutions and balms.

Creams or lotions are oil-in-water emulsions that contain more than 50% water. As an oil-containing base, 15 fatty alcohols are used in the first place, e.g. lauryl, cetyl or stearyl alcohol, fatty acids, e.g. palmitic or stearic acid, liquid to solid waxes, e.g. isopropyl myristate, wool wax or beeswax, and / or hydrocarbons, e.g. petroleum jelly (petrolatum) or paraffin oil. Suitable emulsifiers are surfactants with predominantly hydrophilic properties, such as suitable non-ionic emulsifiers, e.g. fatty acid esters of polyalcohols and ethylene oxide adducts thereof, such as polyglycerol fatty acid esters or polyoxyethylene sorbitan fatty acid esters (Tweens), furthermore polyoxyethylene fatty alcohol ethers or fatty acid esters, or suitable ionic emulsifiers, such as al potassium metal salts of fatty alcohol sulfates, e.g. sodium lauryl sulfate, sodium acetyl sulfate or sodium stearyl sulfate, which are usually present in the presence of fatty alcohols, e.g. cetyl alcohol or stearyl alcohol. Additives to the water phase include agents which prevent the drying of the creams, e.g. polyalcohols, such as glycerol, sorbitol, propylene glycol and / or polyethylene glycols, furthermore preservatives, fragrances, etc.

Ointments or lotions are water-in-oil emulsions containing up to 70%, preferably from about 20% to about 50%, water or aqueous phase. The fatty phase primarily comprises hydrocarbons, e.g. petroleum jelly, paraffin oil and / or hard paraffins, which, to improve the water binding ability, preferably use suitable hydroxyl compounds, such as fatty alcohols or esters thereof, e.g. cetyl alcohol or wool wax alcohol and resp. wool wax. Emulsifiers are suitable lipophilic substances, such as sorbitan fatty acid esters (Spans), e.g. sorbitan oleate and / or sorbitan isostearate. Additions to the water phase include moisture retention agents, such as. 8802.008 polyalcohols, e.g. glycerol, propylene glycol, sorbitol and / or polyethylene glycol, as well as preservatives, fragrances, etc.

Microemulsions are isotropic systems, the basis of which consists of the following four components: water, an emulsifier, such as a ten-side, e.g. eumulgine, a lipid, such as a non-polar oil, e.g. paraffin oil, and an alcohol with a lipophilic group, e.g. 2-octylde cannol. If desired, other additives can be added to the microemulsions.

Fatty ointments are anhydrous and contain, in particular, 10 hydrocarbons, e.g. paraffin, petrolatum and / or liquid paraffins, furthermore natural or partial synthetic fat, e.g. coconut fatty acid triglyceride, or preferably hydrogenated oils, e.g. hydrogenated groundnut or castor oil, further fatty acid partial esters of glycerol, e.g. glycerol mono- and distearate, as well as e.g. the fatty alcohol-increasing fatty alcohols, emulsifiers and / or other additives mentioned in connection with the ointments.

The gels are distinguished between aqueous, anhydrous or water-poor gels, which consist of swellable, gel-forming materials. Firstly, transparent hydrogels based on inorganic or organic macromolecules are used. High molecular inorganic components with gel-forming properties are predominantly water-containing silicates, such as aluminum silicates, e.g. bentonite, magnesium aluminum silicates, e.g. sweeping gum, or colloidal silica, e.g. aerosil. As high molecular organics, natural, semi-synthetic or synthetic macromolecules are used. For example, natural and semisynthetic polymers are derived from polysaccharides with the most diverse carbohydrate building blocks, such as celluloses, starches, tragacanth, gum arabic, agar agar, gelatin, alginic acid and its salts, e.g. sodium alginate, and derivatives thereof, such as lower alkyl cellulose, e.g. methyl or ethyl celluloses, carbonyl or hydroxy-lower alkyl cellulose, e.g. carboxymethyl or hydroxyethyl celluloses. For example, the building blocks of synthetic gel-forming macromolecules are suitably substituted unsaturated aliphates, such as vinyl alcohol, vinyl pyrrolidone, acrylic or methacrylic acid. As examples of such polymers, polyvinyl alcohol derivatives, such as polyviol, polyvinylpyrrolidines, such as collidone, polyacrylates, respectively. polymethacrylates, such as Rohagit S or Eudispert, are mentioned. Conventional additives such as preservatives or fragrances can be added to the gels.

40 Pastes are creams and ointments with secretion-absorbing powder components, such as metal oxides, e.g. titanium oxide or zinc oxide, furthermore talc and / or aluminum silicates, which have the object of binding moisture or secretions present.

Foams are e.g. from pressure reservoirs and are aerosol-form liquid oil-in-water emulsions, wherein halogenated hydrocarbons, such as chlorofluoro-lower alkanes, e.g. dichlorodifluoromethane and dichlorotetrafluoroethane are used as propellants. The oil phase includes hydrocarbons, e.g. paraffin oil, fatty alcohols, e.g. cetyl alcohol, fatty acid esters, e.g. isopropyl-10 myristate, and / or other waxes. The emulsifiers used are, inter alia, mixtures of such having predominantly hydrophilic properties, such as polyoxyethylene sorbitan fatty acid esters (Tweens), and such having predominantly lipophilic properties, such as sorbitan fatty acid esters (Spans). In addition, the usual additives, such as preservatives, etc.

Tinctures and solutions usually contain an ethanolic base, to which water is optionally added and to which, inter alia, polyalcohols, e.g. glycerol, glycols and / or polyethylene glycol, as moisture retention agents to reduce evaporation, and greasers, such as fatty acid esters with low molecular weight polyethylene glycols, ie lipophilic substances soluble in the aqueous mixture as a substitute for the skin-containing ethanol extracted fats and, if necessary, other auxiliaries and additives have been added.

The topically applicable pharmaceutical preparations are prepared in a manner known per se, e.g. by dissolving or suspending the active substance in the base or in part thereof, if necessary. When the active substance is administered as a solution, it is usually dissolved in one of the two phases before the emulsification; when used as a suspension, the active substance is mixed with part of the base after emulsification and then added to the rest of the formulation.

The superiority of the pharmaceutical preparations which can be prepared and formulated according to the invention of the said active compound combination rests on the synergistic antimicrobial action of the individual active compounds.

Synergi sme

In the event that different active ingredients are combined, their properties can be counteracting, additive or also synergistically active. The synergistic effect is apparently the most desirable effect because it allows small doses on the one hand and an improvement in activity over that of the individual components on the other. With the antimicrobial used, in particular the antinykotically active compounds or combinations thereof, the ability of the inhibition of germ growth by measuring the MIC (minimum inhibitory concentration) and the killing percentage is measured by the measurement of the zones of inhibition.

When the expression "Fractional Inhibitory Concentration (FIC)" (fraction inhibition concentration) as fraction 10

Minimum inhibitory concentration (MIC) of the active components in the combination product (test performed with 2 or 3 active components in solution simultaneously) 15 FIC ---

Minimum inhibition concentration (MIC) of the individual active components (the test is carried out with only one of the active components in solution) 20 is defined, SFIC must be equated to the sum of the determined FIC values, if no interaction of the individual components. In other words, if a synergy is present, the sum of the determined FIC values of the individual components is less than the determined quantity SFIC for the combination of the active substances. The determined quantity zFIC provides information about the strength of the interaction of the individual components of a combination.

The minimum inhibitory concentration (MIC) 30 The minimum inhibitory concentration (MIC) in µg / ml for Micona-zol (Miconazole is used as nitrate), salicylic acid, benzoic acid alone or in combination is shown in the following Table A, while the methods leading to the results are described further below.

.8802008 8

Table A

^ emen ^ ί / ug / ml) Fractional Som

Miconazole $ b inhibitory FIC

n1 street conc. (FIC) (partial inhibition ______ concentration) _ 10 C. albicans 4-8 1365 680 0.25 200 - 0.0625 + 0.146 = 0.2 °> 5 - 200 0.125 + 0.294 = 0.389 15 °> 25 100 100 0.063 + 0.07 + 0.15 = 0.28 T. mentagrophytes 1 - 670 310 20 0.5 200 - 0.5 + 0.3 = 0.8 0.06 - 200 0.06 + 0.65 »0 .71 0.06 25 200 0.06 + 0.04 + 0.65 = 0.75 0.25 200 50 0.25 + 0.3 + 0.16 = 0.71 25 S. aureus 4 - S = salicylic acid, B = benzoic acid

The MIC values correspond to expectations. The values for Miconazole show a good effect, while for benzoic acid resp. salicylic acid high concentrations, up to the limit of solubility, are required to achieve a MIC value. Subcultures from wells, which do not show (microbes) growth, show that a concentration of benzoic acid at or near the MIC also had micro-35 bicides, while Miconazole's activity was mainly static, with new growth of microbes occurred on the subcultures.

The determined MIC values of combinations of Miconazole with salicylic acid or benzoic acid or both show higher than only additive * values in antimicrobial activity. The action in the germ Can-40 dida albicans for the combination of the active substances was much better than would be expected by adding the fractionated inhibition concentrations (FIC).

It was observed that concentrations significantly below the MIC values have a significant effect on growth. The observations made here that the individual components all contribute to the antimicrobial activity is significant for the formulated product, the diffusion of the individual components being limited per se.

The method for the determination of the in vitro activities for Micona-10 zol, salicylic acid and benzoic acid in the germs Candida albicans,

Trichophyton mentagraphytes and Staphylococcus aureus are described as follows.

The minimum inhibitory concentration = MIC 15 1. Inoculum C. albicans N.C.Y.C. 610, T. mentagrophytes N.C.P.F. 80 and S, aureus N.C.T.C. 8532 are used as sprouts. C. albicans germs are grown overnight in SLM (Sabouraud liquid medium) at a temperature of 32 ° and the resulting inoculum is diluted to about 20 2 x IQ5 germs based on a total germ count. T. mentagrophytes microconidia germs are withdrawn from a 12 day culture grown on SDA (Sabouraud dextrose agar) at a temperature of 32 ° and a washed aqueous suspension is stored frozen.

The in vitro study is prepared in appropriate dilution in SLM

(Sabouraud liquid medium), as a nutrient medium with germ inoeula of 2 x 105 germs / ml substrate, based on a total germ count. A culture of S. aureus in "Isotonic Sensittest-Broth" (ISB) as a nutrient medium grown at a temperature of 37 ° (overnight) is diluted 1: 500 with ISB. The living germs are then determined by counting.

2. Solutions

By dilution to double, solutions of Micona-zol, salicylic acid and / or benzoic acid in meat broth are prepared. At a higher concentration of salicylic acid and benzoic acid, the concentration of active substance in solution is examined, in which a saturated solution is prepared, which is filtered (for a membrane filter of 0.45 µm. The resulting filtrate, which is used to dilute to achieve double for MIC determination is analyzed by 40 HPLC (high pressure liquid chromatography> 8802008, 10 high pressure) DMF (0.5¾) is used as solvent for Micona-Z and wax present in all environments in this concentration.

3. In vitro research method

The MIC values of the individual active substances are determined by 100-100 µl of volume large wells (wells) being inoculated into a microtiter plate. An equal volume of inoculum is added and the plates incubated at a temperature of 32 ° with both fungal germs and at a temperature of 37 ° with S. aureus. For the MIC values, combinations of two compounds 10 each are prepared and therefrom three different required concentrations of solutions, and then 75 µl of each compound and of each inoculum are fed to the wells of the microtiter plate and incubated as described . A three-component system is also being investigated, using 50 µl volume to inoculate the plates.

Results

The results are read 24 hours after incubation (incubation) for C. albicans and S. aureus and 6-7 days for T. mentagraphytes. The end points are determined visually and by measuring the "optical density" 20 using a vertical raster photometer at 450 nm. In particular, the optical density reading makes it possible to investigate the effect on growth of concentrations below MIC values.

The antimicrobial activity of various cream formulations is investigated by estimating the zones of inhibition by the following methods.

Measurement of the braking zones (spread plate)

About 1 x 104 seeds / ml of each seed to be examined are inoculated (inoculated) onto the surface of an agar plate (SDA for yeast and fungal germs and 30 TSA for bacteria) and dried for 30 minutes. Floors (wells) are made in the agar of the plate with a cork drill number 3 and filled with about 100 mg of cream. The plates are incubated overnight (with fungal germination longer) at a temperature of 32 ° and the diameter of the inhibition zones is measured (see table).

Germs applied:

Candida albicans, Trichophyton mentagrophytes, Staphylococcus aureus, Pseudomonas aeruginosa.

A comparison of all agar diffusion tests shows that the combination MSB (Miconazole + salicylic acid + benzoic acid) is significantly more effective than the others, namely Miconazole alone or the combination SB (salicylic acid + benzoic acid).

Summarized results of the cream formulations MSB = cream, containing Miconazole, salicylic acid and benzoic acid 5 SB = cream, containing salicylic acid and benzoic acid D = Daktarirfè * (2¾ Miconazole) (cream prepared by Janssen Pharmaceuticals, Belgium) M = Miconazole 2¾.

10 Table B

Organisms (germs) Cream formulas

Braking zones (diameter, mm)

_MSB SB_M_D

15 C. albicans (NCYC610) (1) 27 17.7 9.2 17.2 (2) 33.7 24.7 16.7 39.8 (3) 29.3 * 11.3 24.0 T. mentagrophytes 55 38 30.5 43 20 NC0F80, pre-germinated T. mentagrophytes (1) 54.3 52.5 37.8 43 NCPF80, microconidia (2) 49.0 55.3 45.3 40.8 (3)> 80 * 43.5 54 25 T. mentagrophytes 52.3 50.3 41.8 37 ATCC 18748 (spread plate results) 30 T. mentagrophytes 56 54.6 28 * C. albicans 24 20 15.6 * 35 T. mentagrophytes 44 , 6 34.9 * 30.9 MSB v D and M is significant at 0.05¾ (level) MSB V SB is significant at 2.5¾ (level) * = not measured 40 t 8802008 12 'Diffusion studies

Most pharmaceutical preparations consist of the active ingredient and excipients and it is therefore extremely important to investigate the activity of the compound (active ingredient) in the chosen formulation. For topical applicability, the diffusion of the active substance resp. active substances from the formulation must be determined and for this purpose the plate diffusion test is used to investigate the applicability of the product. The results are shown in Table C. Formulations 1 to 7 all contain the same base composition, however the active ingredients are different. The abbreviations used are self-evident, for example M2S3 stands for 2% Miconazole and 3% salicylic acid. DJ stands for DaktarirJ§> Janssen, AJ stands for Acnidazil® Janssen. As you go through the data, it should be noted that the microorganisms (germs) used are the most suitable combination of Miconazole, benzoic acid and salicylic acid on average. It is noted that the comparisons of all formulations are performed at the same time, because comparisons on different days do not give the same results. The relative ranking in itself remains the same.

t 8802008 13

Determination of the antinricrobial activity of a cream formulation using a variety of germs

5 Table C

Sprouting Inhibition zones (diameters mm = of the cream formulation) _1. 2. 3. 4. 5. 6. 7. 8. 9.

M2S3 M2 S3B6 S3 B6 M2B6 M2S3B6 DJ AJ 10 (C. albicans) 15.3 9.2 17.7 0 11.2 23 27 17.2 * (NCYC610) T. mentagrophytes (NCPF80) 34 30.5 38 27.3 37.3 47.5 55 43 * 15 pre-germinated T. mentagrophytes (NCPF80) 31.5 37.8 52.5 22.5 42 51.3 54.3 43 * microconidia 20 S. aureus 49.3 24.5 42.8 43.2 34.2 36 48.9 30.2 20.3 (NCTC6749 SDA plates)

Ps. aeruginosa 11.8 0 21.3 14.3 14.8 16.7 20.8 12.7 0 25 (NCTC10332 SDA plates) C. albicans 33.3 16.7 24.7 11.2 21.5 33 .3 33.7 39.8 20.3 (NCYC610) 30 T. mentagrophytes 50.5 41.8 50.3 28.3 47.8 54.2 52.3 37 32 (ATCC18748) T. mentagrophytes 61 .8 45.3 55.3 36.8 45.8 60.3 49.0 40.8 33 (NCPF80) 35 * = not measured M = Miconazole (%), B = benzoic acid (%) S = salicylic acid ( %), DS = DaktarirÊ) (Janssen), AJ = Acnidazole © (Janssen) 40 i8802008 14

Examined Cream Formulations (Prepared and Described in Examples IV-VI)

Cream 1

Composition 1% 5 soft paraffin (white) 23.75 propylene glycol 11.40 cetostearyl alcohol 23.75 cetrimide 0.95 methyl paraben 0.025 10 propyl paraben 0.015 water 35.15

Miconazole nitrate 2.00 salicylic acid 3.00 benzoic acid 0.00 15

Cream 2

Composition% soft paraffin (white) 24.50 propylene glycol 11.75 20 cetostearyl alcohol 24.50 cetrimide 0.98 methyl paraben 0.025 propyl paraben 0.015 water 36.25 25 Miconazole nitrate 2.00 salicylic acid 0 benzoic acid 0

Cream 3 30 Composition 1% soft paraffin (white) 22.75 propylene glycol 10.92 cetostearyl alcohol 22.75 cetrimide 0.91 35 methyl paraben 0.025 propyl paraben 0.015 water 33.67

Miconazole nitrate 0.00 salicylic acid 3.00 40 benzoic acid 6.00 i8802008 15

Cream 4

Composition% soft paraffin (white) 24.25 propylene glycol 11.64 5 cetostearyl alcohol 24.25 eatrimide 0.97 methyl paraben 0.025 propyl paraben 0.015 water 35.89 10 Miconazole nitrate 0.00 salicylic acid 3.00 benzoic acid 0.00

Cream 5 15 Composition% soft paraffin (white) 23.50 propylene glycol 11.28 cetostearyl alcohol 23.50 appeti 0.94 20 methyl paraben 0.025 propyl paraben 0.015 water 34.78

Miconazole nitrate 0.00 salicylic acid 0.0025 benzoic acid 6.00

Cream 6

Composition% soft paraffin (white) 23.00 30 propylene glycol 11.04 cetostearyl alcohol 23.00 edrimide 0.92 methyl paraben 0.025 propyl paraben 0.015 35 water 34.04

Miconazole nitrate 2.00 salicylic acid 0.00 benzoic acid 6.00. 8802 008.

16

V

Cream 7

Composition 1 ng% soft paraffin (white) 22.25 propylene glycol 10.68 5 cetostearyl alcohol 22.25 cetrimide 0.89 methyl paraben 0.025 propyl paraben 0.015 water 32.93 10 Miconazole nitrate 2.00 1) salicylic acid 3.00 benzoic acid 6.00

Cream 8 DaktariilD (Janssen) 2 wt.% Miconazole nitrate.

15

Cream 9 Acnidazill) (Janssen) 2 wt.% Miconazole nitrate and 5 wt.% Benzoyl peroxide.

1) The concentration was reduced to 0.1¾, with no loss of activity detectable.

Fungicidal activity of Miconazole with salicylic acid and / or benzoic acid Subcultures of MIC tests performed for the determination of the MBC (minimum bactericidal concentration = minimum bactericidal concentration) indicate that benzoic acid and salicylic acid have a fungicidal activity at concentrations that only slightly higher than the concentrations of the determined MIC values. The microbicidal activity of active ingredients was assayed by micro-crocididia using the method of determination of the kill rate.

Concentrations of 20 20 µg / ml of Miconazole were fungicidal within 24 hours and benzoic and salicylic acid at concentrations of 400-600 µg / ml.

A combination of further lower concentrations over a 6 hour test period shows that salicylic acid increases the fungicidal rating of Miconazole at a concentration of 20 or 10 µg / ml each time. The combination of benzoic acid and salicylic acid again gives an improved fungicidal effect.

The activity against tracks can be classified as both inhibitory and d-40 odend. The proof is provided because one. 8802008 17 in the presence, respectively. after removing the active ingredients, by measuring the ability to form colonies, the activity determines.

Miconazole (10 µg / ml) and salicylic acid (200 µg / ml) alone or 5 combinations of both have an inhibitory effect, because germination is inhibited, however some germs survive.

Efficacy of salicylic acid and miconazole by demonstrating viability and germination capacity (%) of T. mentagrophytes microconidia 10

Table D

Combination Vascular (cfu / ml)% germination pH

feasibility at 15 at time (h) time (h) _6_24 _6 24_ control 3.5 x 105 2.4 x 105 12 65 M10 5.9 x 105 7.0 x 104 9 11 5.88 M10 / S 200 8 , 3 x 10 * 2.3 x 10 ^ 6 16 5.41 20 M10 / S 400 0 0 4 13 5.06 M20 3.3 x 1Q5 4.6 x 105 7 5 5.84 M20 / S 200 2, 2 x 104 5.3 x 103 8 13 5.41 M20 / S 400 0 0 9 12 5.07 S 200 4.8 x 105 1.2 x 105 12 41 5.43 25 S 400_0_0_14 14 5.08 M = Miconazole, S = salicylic acid

Initial count at T0 gives 7.9 x 105 cfu / ml. Different concentrations of Miconazole in the cream formulations are investigated in T. menta-30 grophytes (Table E). The formulations containing MSB are significantly better than those containing SB. The 2% MSB-containing cream formulation was significantly better than the others tested, whereas those containing 0.1-1% Miconazole are not significantly different from each other, in particular, however, significantly better than formulations containing SB, or commercial products containing only 2% Miconazole (Daktarir®).

v 8802008 4 18

Effect of the concentration of Miconazole in cream formulations containing salicylic acid (3%) and benzoic acid (6%) against T. mentagrophytes

5 Table E

Cream Diameter braking zones (mm) _1_2_3_4_5 average. St.afw.

1. Miconazole 2% 46 44 45.5 45 42.5 44.6 1.39 10 2. Miconazole 1% 40 40.5 43 44.5 43.44 42.2 1.89 3. Miconazole 0.5¾ 40 .5 40.5 40.5 44 42 41.8 1.44 4. Miconazole 0.25% 42.5 40.5 40 42 39 40.8 1.44 5. Miconazole 0.1% 42.5 40, 5 40.5 40 42.5 41.2 1.20 6. Miconazole 0% 34 36 35.5 35 34 34.9 0.89 15 7. Miconazole 2% 33.5 30 31 29 NT 30.9 1. 93 (Daktarin) _ T. mentagrophytes 7.5 x 104 Significance (0.05%) 20 Multiple-Range Test (Neuman Keuls) for Significance (0.05%) 1 vs 2,3,4,5,6 , 7 significant differences 2 vs 3,4.5 geert significant differences 2 vs 6.7 significant differences 25 3 vs 4.5 no significant differences 3 vs 6.7 significant differences 4 vs 5 no significant differences 4 vs 6.7 significant differences 5 vs 6.7 significant differences 30 6 vs 7 significant differences

Table E shows that the synergistic effect can be obtained over a wide range of different concentrations of Miconazole.

35 2. Further evidence of synergy can be provided from the kinetic data in Table F.

The death rate of T. mentagrophytes microconidia when Miconazole (M) alone or in combination with benzoic acid (B) and salicylic acid (S) is determined.

.8802008 19

Table F

Time M20 M20 + S 200 M20 + B 200 M10 M10 + S 200 M10 + B 200 5 yh) _ 1 1 xl05 1.1x105 6.7x104 2.0x105 1s3x105 1.2x105 2 3.3x104 1.2x104 2.3x104 9 , 2x104 5.8x104 4.2x104 10 4 2.2x10-4 Ι, δχΙΟ3 1.9x104 Ι, βχΙΟ3 1.5x10 * 2.5xl04 6 1.4x104 5.0x102 1.2x1 <) 4 7.5x104 8.3xl03 2.6χΠ) 4 24 8.3x102 o 4.2x102 2.3x10 * 153x102 1.7x10 * pH values in advance 5.98 5.57 5.50 5.94 5.54 5.49 afterwards 5.95_5.54_5 , 50_5,93_5,53_5,48 S = salicylic acid, B = benzoic acid

Initial count at T0 gives 5.9 x 103 efu / ml.

All solutions are run in SLM * (SLM + 0.5% DMF) on the previous day to ensure a complete solution.

To 9 ml of Trichophyton mentagrophytes microconidia, 9.9 ml of a sample (estimated T. ment. At 1 x 108 cfu / ml) is added. The counting of the viable germs takes place from a germination solution. The withdrawn volumes of 0.1 ml are performed with a 100 µl pipette.

The samples are kept at a temperature of 32 ° C without exception.

1 ml is removed after 1, 2, 4, 6, 24 hours and the viable germ counting is performed.

Serial dilutions are in 0.1% peptone water and are plated using the Miles and Migra method on SDA (Sabouraud dextrose agar). The plates are read after 6 and 14 days.

.8802008 t 20

Sample Concentration 1 M 20 / µg / ml 2 M20 + S 200 / µg / ml 53 M 20 + B 200 / µg / ml 4 M 10 / µg / ml 5 M 10 + S 200 / µg / ml 6 M 10 + B 200 µg / ml 10 S = salicylic acid B = benzoic acid MIC values are also performed in (SLM *) for Miconazole.

Miconazole concentrations are: 4, 2, 1, 0.5, 0.25, 0.125 µg / ml (duplicates).

0.1 ml of T. microconidia is added to obtain a final concentration of about 1 x K) 5 cfu / ml (the same germination beginning as above).

The invention is illustrated by, but not limited to, the following examples.

Example I

A powder containing 2% Miconazole nitrate, 2% salicylic acid and 3% benzoic acid is prepared in the following manner:

Composition 1i ng 25 Miconazole nitrate 2% salicylic acid 2% benzoic acid 3% amorphous silica 0.1% talc, purified and finely ground ad 100% 30

Miconazole nitrate, salicylic acid and benzoic acid are finely ground in the presence of amorphous silica. The microfine mixture is then added in small amounts to the sterile, finely ground and purified talc. Mixing is continued until a homogeneous product is obtained.

Example II

An aerosol formulation containing 3% Miconazole nitrate, 3% salicylic acid and 6% benzoic acid is prepared as follows:. 8802008 21

Composition

Mi conazole nitrate 3% salicylic acid 5% 5 benzoic acid 6%

Tween 80 0.1% amorphous silica 0.1% deodorized hydrocarbon s.q.

A finely ground mixture of Miconazole nitrate, salicylic acid, benzoic acid and amorphous silica is prepared (as described above in the powder preparation). Tween 80 is then sprayed as an alcoholic solution at intervals to ensure evaporation of the support material and even distribution. The Tween 80 coated powder is then slurried in the deodorized hydrocarbon and then packaged under normal standard techniques. A non-flammable with suitable properties is selected as the hydrocarbon.

Example III

A lotion formulation containing 1% Miconazole nitrate, 3% salicylic acid and 6% benzoic acid is prepared as follows:

Composition

Miconazole nitrate 1% 25 salicylic acid 3% benzoic acid 6% butylhydroxytoluene 2% diethyltoluamide 2% polyethylene glycol 400 ad 100% 30

Miconazole nitrate, salicylic acid and benzoic acid are dissolved in half of the total amount of polyethylene glycol 400 used and butylhydroxytoluene and diethyltoluamide are added until a clear solution is obtained. The remaining polyethylene glycol 35 400 is then added for backfilling.

Example IV

A cream containing 2% Miconazole nitrate, 3¾ salicylic acid and 6% benzoic acid is prepared as follows:. 8802008 o 22

Composition Soft paraffin (white) 22.25% proplylene glycol 10.68% 5 cetostearyl alcohol 22.25% cetrimide 0.89% methyl paraben 0.025% propyl paraben 0.015% water 32.93% 10 Miconazole nitrate 2.00% salicylic acid 3.00% benzoic acid 6.00%

A melting or dispersion technique can be used for the preparation. The white soft paraffin, propylene glycol and the cetostearyl alcohol are mixed and heated until an even viscous liquid is obtained. Miconazole nitrate, salicylic acid and benzoic acid are dissolved with paraben in this liquid. The cetrimide is dissolved in water at 60 ° and added to the oil phase with continuous stirring. A homogeneous cream is obtained. The remaining water is added and the mixture is further homogenized.

Example V

A cream formulation containing 2% Miconazole nitrate and 3% salicylic acid is prepared as described in Example IV: 25

Composition soft paraffin (white) 23.75% propylene glycol 11.40% cetostearyl alcohol 23.75% 30 cetrimide 0.95% methyl paraben 0.025% propyl paraben 0.015% water 35.15%

Miconazole nitrate 2.00% 35 salicylic acid 3.00%

Example VI

A cream formulation containing 2% Miconazole nitrate and 6% benzoic acid is prepared as described in Example IV: 8802008 23

Composition soft paraffin (white) 23.00% proplylene glycol 11.04% 5 cetostearyl alcohol 23.00% cetrimide 0.92% methyl paraben 0.025% propyl paraben 0.015% water 34.04% 10 Miconazole nitrate 2.00% salicylic acid 0.00% benzoic acid 6 .00%

Example VII

15 A cream containing 2% CTotrimazole, 3% salicylic acid and 6% benzoic acid is prepared as follows:

Composition soft paraffin (white) 22.25% 20 propylene glycol 10.68% cetostearyl alcohol 22.25% cetrimide 0.89% methyl paraben 0.025% propyl paraben 0.015% 25 water 32.93% CTotrimazole 2.00% salicylic acid 3.00% benzoic acid 6 Analogous to Example IV, a melting or dispersion technique can be used for the preparation. The white soft paraffin, propylene glycol and the cetostearyl alcohol are mixed and heated until an even viscous liquid is obtained. CTrimazole, salicylic acid and benzoic acid are dissolved in this liquid with paraben and added to the oil phase with continuous stirring. A homogeneous cream is obtained. The remaining water is added and the mixture is further homogenized.

Example VIII

A cream formulation containing 2% Clotrimazole and 3% salicylic acid is prepared as described in Example VII:> 8802008

V

\ 24

Composition soft paraffin (white) 23.75% proplylene glycol 11.40% 5 cetostearyl alcohol 23.75% edrimi de 0.95% methyl paraben 0.025% propyl paraben 0.015% water 35.15% 10 Clotrimazole 2.00% salicylic acid 3.00%

Example IX

A cream formulation containing 2% Clotrimazole and 6% benzoic acid 15 is prepared as described in Example VII:

Composition soft paraffin (white) 23.00% propylene glycol 11.04% 20 cetostearyl alcohol 23.00% edrimi 0.92% methyl paraben 0.025% propyl paraben 0.015% water 34.04% 25 Clotrimazole 2.00% salicylic acid 0.00% benzoic acid 6.00% 8802008

Claims (11)

1. Antimicrobial pharmaceutical preparations for the topical application, characterized by the combination, which consists of an anti-microbial active substance from the group of active substances, which in its molecule contains an imidazole substituted with a lower alkyl, corresponding to a compound of formula 1, wherein at least one of the substituents R 1, R 2 and R 3 represent a halogen atom and the other hydrogen, as well as its pharmaceutically acceptable acid addition salts or corresponding to a compound of formula 2, wherein R 4 represents a halogen atom, with one or more optionally substituted benzoic acids and optionally conventional pharmaceutical carriers and / or excipients. Antimicrobial pharmaceutical preparations for the topical use according to claim 1, characterized by the combination consisting of a compound of the formula 1, in which at least one of the 15 substituents R 1, R 1 and R 3 represent a chlorine atom and the remaining hydrogen, as well as its pharmaceutically acceptable acid addition salts, such as e.g. Miconazole (Ri = R2 = Cl, R3 = H), Econazole (R3 = Cl, Ri = R3 = H) and Isoconazole (Ri = R3 = Cl, R2 = H) or from a compound of formula 2, wherein R4 is 20 means chlorine atom, such as e.g. Clotrimazole and one or more optionally substituted benzoic acids and optionally conventional pharmaceutical carriers and / or excipients. Antimicrobial pharmaceutical preparations for the topical use according to claim 1 or 2, characterized by the combination consisting of Miconazole as a compound of the formula 1, wherein R 1 and R 2 represent a chlorine atom and R 3 hydrogen or of a pharmaceutically acceptable acid addition zone or from a compound of the formula II, wherein R4 represents a chlorine atom, Clotrimazole and salicyclic acid and / or benzoic acid and optionally conventional pharmaceutical carriers and / or excipients. Antimicrobial pharmaceutical preparations for the topical use according to any one of the preceding claims, characterized by the combination consisting of Miconazole nitrate and salicylic acid and / or benzoic acid and optionally conventional pharmaceutical carriers and / or auxiliary substances. Antimicrobial pharmaceutical preparations for the topical use according to any one of the preceding claims, characterized by the combination consisting of Clotrimazole and salicylic acid and / or benzoic acid and optionally conventional pharmaceutical carriers and / or excipients. <. 8802008 V Antimicrobial pharmaceutical preparations for the topical use according to any one of the preceding claims, characterized in that the preparations are 0.01-10% by weight of an imidazole of the formula 1 or 2 and substituted with a lower alkyl group and 0.5- 25% by weight of one or more optionally substituted benzoic acids together with conventional carriers and / or excipients for the topical application. Antimicrobial pharmaceutical preparations for the topical use according to any one of the preceding claims, characterized in that the preparations comprise 0.1-2.0% by weight of an imidazole of the formula 1 or 2 substituted with a lower alkyl group and 1.0-15.0% by weight of one or more optionally substituted benzoic acids together with conventional carriers and / or excipients for the topical application. Antimicrobial pharmaceutical preparations for the topical use according to any one of the preceding claims, characterized in that the preparations comprise a synergistically effective amount of an imidazole of the formula 1 or 2 substituted with a lower alkyl group and one or more optionally substituted benzoic acids together with conventional carriers and / or excipients for the topical application. Antimicrobial pharmaceutical preparations for the topical application according to any one of the preceding claims, characterized in that the preparations are in the form of a gel, a cream, an ointment, a paste, a powder, a solution, a tincture, a foam, a spray, aerosol, collodion or lotion. 10. A method of preparing an antimicrobial pharmaceutical composition for the topical application, characterized in that an antimicrobial active substance from the group of active substances, which in its molecule contains an imidazole substituted with a lower alkyl, or a compound of the formula 1, wherein at least one of the substituents R 1, R 2 and R 3 represent a halogen atom and the other hydrogen, as well as its pharmaceutically acceptable acid addition salts, or corresponding to a compound of formula 2, wherein R4 means a halogen atom with one or more optionally substituted benzoic acids and optionally mixes conventional pharmaceutical carriers and / or auxiliaries. Use of an antimicrobial pharmaceutical preparation according to any one of claims 1 to 9 as an antimycotic active agent for the treatment of skin infections caused by dermatophytes and Brussels sprouts, as an antibacterial active agent for the treatment of acne, dandruff and other skin diseases and as an anti-pruritic active agent for the treatment of pruritus diseases. +++++++ v 8802008