Pharmaceutical carriers for active substances and pharmaceutical compositions containing said carriers

Abstract

The present invention relates to a pharmaceutical carrier for active substances, characterized in that it contains a strongly solvating aprotic solvent from the group of sulfoxides, a carbonic acid amide, and a vegetable oil with a high content of unsaturated fatty acids, as well as to pharmaceutical compositions containing this carrier.

Description

[Technical field]

[0001] The present invention relates to a semi-solid drug form containing a carrier for an active substance. The present invention is applicable in pharmacy and medicine. [Background technology]

[0002] Water-soluble and other lipid-soluble active substances are known, which may be administered in parallel in some disease conditions involving inflammation. Inflammation is an ordered process that occurs in vascularized tissues under the influence of injury factors. Inflammation can be caused by chemical, physical or biological (infection with pathogenic microorganisms) factors, which are generally exogenous or endogenous factors. Changes in blood vessels are at the root of inflammatory reactions. Various plasma proteins with defensive functions, such as antibodies or complement, can enter the affected tissue, resulting in vasodilation, an increase in blood supply to the tissue and vascular permeability.

[0003] Among these, the active substance is Curcuma longa (Curcuma domestica). It is a perennial plant of the ginger family, cultivated mainly in the equatorial zone for its flavor. The herbal raw material is long curcuma (Curcumae longae rhizoma), which contains at least 3% dicinoylmethane derivatives (curcuminoids) calculated as curcumin and at least 3% oil (mainly sesquiterpenes) based on the dry raw material. Curcumin has been shown to inhibit TPA-induced inflammation, hyperplasia and proliferation. Turmeric rhizome inhibited the inflammatory response caused by the use of bleomycin and amiodarone in rats and prevented pulmonary fibrosis. The anti-inflammatory effect of turmeric is related to the inhibition of arachidonic acid metabolism in mouse epithelium. Studies have shown that the average effective dose of curcumin in the carrageenan test was ED50=48mg / kg in rats and ED50=100.2mg / kg in mice. ED50=309.0mg / kg for alcoholic extract of turmeric, ED50=4.7mg / kg for aqueous extract, ED50=40.7mg / kg for petroleum ether extract, ED50=8.7mg / kg for curcumin alone in mice. Curcuma longa rhizome can be used to treat several chronic inflammatory diseases. Nahar et al. carried out experiments on a preparation containing a solid complex of lipids with curcumin. The in vitro activity of this preparation against lipopolysaccharide (LPS)-induced inflammation was tested in the RAW 264.7 mouse macrophage line. The use of the preparation significantly reduced the concentrations of nitric oxide and prostaglandin E2 as well as interleukin-6 in a dose-dependent manner. Inhibition of the transcription factor NF-κB has also been observed. Published results of pharmacological studies have shown the inhibitory effect of turmeric components on the activity of cyclooxygenase in mouse epidermal microsomes and lipoxygenase in the cytosol. In the Ames test, both the aqueous extract of turmeric and isolated curcumin were non-mutagenic.

[0004] Coptis is used in traditional Japanese, Chinese and Indian medicine to treat numerous diseases. Considering the pharmacological properties of the described isoquinoline alkaloid raw material, it can give the following range of actions: dilating, choleretic, protozoicidal, bacteriostatic and bactericidal, fungistatic, hypotensive, sedative, antipyretic, analgesic, cytotoxic, anthelmintic. Coptis inhibits the release of toxins by bacterial cells. Coptis limits the in vitro growth of staphylococci, streptococci, pneumococci, Vibrio cholerae, Bacillus anthracis and Dysenteriae Bacillus. Coptis preparations are successfully used in the treatment of leishmania infections (leishmaniasis). Coptis extract, like other herbs containing berberine, destroys numerous pathogenic protozoa, such as Entamoeba histolytica, Giardia lamblia, and Trichomonas vaginalis. Some authors (e.g. John Chen and Tina Chen, 2004) have reported that Coptis has a very broad spectrum of protection against: Bacillus dysenteriae, Mycobacterium tuberculosis, Salmonella typhi, E. coli, Vibrio cholerae, Bacillus proteus, Pseudomonas aeruginosa, Diplococcus meningitid, Staphylococcus aureus, beta-hemolytic streptococcus, Diplococcus pneumoniae, Corynebacterium diphtheriae, Bordetella pertussis, Bacillus anthrax, and Bacillus It has been reported that it has antibacterial activity against Bacillus anthracis and Leptospira.They believe that Coptis significantly inhibits the development of E. coli, Mycobacterium tuberculosis and Staphylococcus aureus.Coptis Chinensis external preparations are indicated for the treatment of inflammation of the mouth and throat, inflammation of the skin, infections of the skin and mucous membranes of the genitals, conjunctivitis and eyelids, eye parasitosis, psoriasis, dandruff, proctitis, vaginal discharge as a result of genital tract infections, mycoses of the skin and mucous membranes (strong exudates and aqueous-alcoholic extracts), boils, purulent skin diseases, acne. Summary of the Invention [Problem to be solved by the invention]

[0005] The carrier is therefore required to ensure the dissolution or emulsification of both water-soluble and fat-soluble active substances, and at the same time to increase their bioavailability, since the dosage of these substances can be reduced while increasing their activity, as well as to make it possible to obtain semi-solid mixtures and compositions containing them with high homogeneity and uniformity and low levels of impurities.Surprisingly, said object has been solved by the present invention. [Means for solving the problem]

[0006] A first object of the present invention is a pharmaceutical carrier for active substances, characterized in that it contains a strongly solvating aprotic solvent from the group of sulfoxides, a carbonic acid amide, and a vegetable oil with a high content of unsaturated fatty acids.Preferably, the carrier according to the present invention is characterized in that the strongly solvating aprotic solvent from the group of sulfoxides is selected from the group comprising dimethyl sulfoxide.Equally preferably, the carrier according to the present invention is characterized in that the carbonic acid amide is selected from the group of urea, caffeine or absorption enhancers comprising caffeine.In a further preferred embodiment of the present invention, the carrier is characterized in that the vegetable oil with a high content of unsaturated fatty acids is selected from the group comprising dandelion oil, sunflower oil, vegetable oil, preferably linseed oil, rapeseed oil, peanut oil, hemp seed oil.In another equally preferred embodiment of the present invention, the carrier is characterized in that it additionally comprises a natural emulsifier, preferably selected from the group comprising white wax, vegetable wax, preferably yellow wax, carnauba wax, lanolin. In another equally preferred embodiment of the invention, the carrier is characterized in that it contains a substance from the group of sulfoxides as a strongly solvating aprotic solvent in an amount of 0.01% to 5% by weight of the drug. More preferably, the carrier according to the invention is characterized in that it contains a carbonic acid amide in an amount of 0.01% to 2% by weight of the drug. Equally preferably, the carrier according to the invention is characterized in that it contains a vegetable oil with a high content of unsaturated fatty acids in an amount of 1% to 5% by weight of the drug. Most preferably, the carrier according to the invention is characterized in that it contains a natural emulsifying substance in an amount of 0.001% to 8% by weight of the drug.

[0007] A second object of the present invention is a pharmaceutical composition comprising a carrier and an active substance as defined in the first object of the present invention, characterized in that the ratio of active substance to carrier is between 1:10 and 2:1, the carrier constituting between 1% and 20% by weight of the composition. Equally preferred, the composition according to the present invention is characterized in that it comprises a matrix, the weight ratio of active substance to excipients is between 1:2.5 and 1:494, and preferably has a semi-solid drug form. More preferably, the composition according to the present invention is characterized in that it comprises an active substance of natural or synthetic origin, preferably selected from the group containing Coptis Chinesis extract, Curcuma Longa extract or combinations thereof. In another preferred embodiment of the present invention, the composition is characterized in that the matrix component is selected from the group containing lard, more preferably duck lard and / or goose lard and / or Eucerin, more preferably Lekobaza, Hascobaza, white petrolatum, yellow petrolatum and / or zinc sulfate-zinc salts, any combination thereof, most preferably lard, goose fat, Eucerin and zinc sulfate.

[0008] The mixture of excipients creates a carrier characterized by its effect on the bioavailability of the active substance. For example, a semi-solid carrier of the active substance is made using the following excipients: A, preferably dimethyl sulfoxide (DMSO), B, preferably urea (urea), D, Oleum Chaulmoogra (Oleum Chaulmoogra), plus C, preferably white wax (Cera alba).

[0009] The proportions of the carrier components are shown in Table 1.

[0010] [Table 1]

[0011] The mixture of substances constituting the carrier consists of elements that ensure the dissolution or emulsification of both water-soluble and lipophilic active substances. Mixture AB is responsible for the dissolution of hydrophilic active substances. Mixture AD, preferably in addition C, is responsible for the dissolution of lipophilic substances. Mixture AB, preferably in addition C, is responsible for emulsifying properties. The minimum amount of A used in the carrier must not be lower than 0.01% of the total weight of the drug and must not exceed 5% of the total weight of the drug. · The minimum amount of B used in the carrier must not be lower than 0.01% of the total weight of the drug and must not exceed 2% of the total weight of the drug. The minimum amount of C used in the carrier should not be lower than 0.001% of the total weight of the drug, and should not exceed 8% of the total weight of the drug, and no compound of group C needs to be used in the basic carrier of the present invention. · The minimum amount of D used in the carrier must not be lower than 1% of the total weight of the drug and must not exceed 5% of the total weight of the drug.

[0012] The goal of the present invention is to obtain a universal carrier for active substances, which allows the dose of these substances to be reduced while increasing their activity, thus increasing their bioavailability. The effect is to accelerate the treatment of inflammation and shorten the treatment of skin ulcers.

[0013] The current state of knowledge indicates that mixtures containing no ABD, preferably in addition no C, in particular oleander oil, DMSO, urea and white wax, have been used for this purpose.

[0014] 1 illustrates an exemplary embodiment of the present invention. [Brief description of the drawings]

[0015] [Figure 1] FIG. 2 shows the activity of medicinal products containing substances from the group of sodium-glucose cotransporter 2 (SGLT2) inhibitors (fluorescein). [Diagram 2]FIG. 1 shows a positive response in the form of green emission in muscles adjacent to the fluorescent dye injection site and an absence of positive response in the F2 group (fluorescein). [Diagram 3] FIG. 1 illustrates a sample of hepatocytes at 400× magnification with a strong positive response in group F3 (complete staining of hepatocytes and intercellular spaces) and a negative response in the remaining groups (fluorescein). [Figure 4] FIG. 1 illustrates at 40x magnification that a strong positive response in the form of green luminescence in the dermis and subcutaneous layer was strongest in F1 and F3 groups (arrows), with a positive response occurring only on the surface of the epidermis in F3 group (acridine orange). [Diagram 5] Figure 1 shows muscle tissue where a positive response was visible, i.e. similar to that seen in skin, and image contrast had to be corrected to show the signal at low magnification (acridine orange).Negative responses were observed in the F2 group (strong image contrast enhancement, magnifications 100x, 100x and 400x). [Figure 6] FIG. 1 shows hepatocyte samples at 400× magnification where a positive response occurred in the F3 group (arrows) while negative results were observed in the other groups (acridine orange). [Figure 7] Illustrated samples of connective tissue cells at 100x, 100x and 40x magnifications (Rhodamine), with the strongest positive responses observed in groups F1 and F3. [Figure 8] A weak positive response is visible in groups F1 and F3 (magnification 100x), picture of liver samples (rhodamine). [Figure 9] Normalized value plot in which the mean fluorescence intensity of test tissues is a percentage of the fluorescence intensity of control tissues treated with vehicle without antibody (100%). [Figure 10] FIG. 1 shows absolute mean fluorescence intensity values ​​of test formulation B in the dermis. [Figure 11] A plot of normalized fluorescence intensity in the dermis compared to vehicle control (100%). [Figure 12]FIG. 1 illustrates the nonparametric Kruskal-Wallis ANOVA ("z" value for multiple comparisons; W, independent variable (grouping): S; Kruskal-Wallis test: H(7, N=24)=14.69333, p=.0401) of the comparison results of all tested samples obtained. [Figure 13] FIG. 1 shows the results of the study described in Example 35. [Figure 14] FIG. 1 shows the results of the study described in Example 35. Example 1

[0016] A carrier mixture (F1) was prepared in the following steps:

[0017] a) A vegetable oil with a high content of unsaturated fatty acids and optionally a natural emulsifying agent are melted in a vessel at a maximum of 40°C.

[0018] b) A strongly solvating aprotic solvent from the group of sulfoxides was added, with stirring, to a molten mixture of vegetable oil with a high content of unsaturated fatty acids and optionally a natural emulsifying substance.

[0019] An appropriate amount of carbonic acid amide was added to a mixture of a strongly solvating aprotic solvent from the group of sulfoxides, vegetable oil D with a high content of unsaturated fatty acids, and optionally a natural emulsifying substance.

[0020] The resulting mixture was stirred until a homogenous consistency and color was achieved.

[0021] The resulting mixture exhibited emulsifying properties. Hydrophilic substances were dissolved in the molten mixture. Lipophilic substances were dissolved in the mixture at room temperature.

[0022] In a subsequent step, mixtures F2 and F3 were prepared. The composition of the mixtures used in the tests is listed in Table 2.

[0023] [Table 2]

[0024] Mixture F2 did not have a strongly solvating aprotic solvent (DMSO) from the group of sulfoxides. The mixture showed emulsifying properties. Lipophilic substances dissolved in the mixture at room temperature. Hydrophilic substances did not dissolve in mixture F2.

[0025] Mixture F3 did not have a natural emulsifier (beeswax). Lipophilic substances were dissolved in the mixture at room temperature. Hydrophilic substances were dissolved in the F3 mixture at about 40° C. Mixture F3 did not show emulsifying properties.

[0026] Tests showed that the F1 mixture had the greatest universal properties. Example 2

[0027] The carrier mixture (F1) was prepared in the following steps:

[0028] a) A vegetable oil having a high content of unsaturated fatty acids in an amount of 1% of the total weight of the drug, and optionally a natural emulsifying agent in an amount of 0.001% of the total weight of the drug, were melted in a vessel at a maximum of 40°C.

[0029] b) A strongly solvating aprotic solvent from the group of sulfoxides in an amount of 0.01% of the total drug weight was added, with stirring, to a molten mixture of a vegetable oil with a high content of unsaturated fatty acids in an amount of 1% of the total drug weight and, optionally, a natural emulsifying substance in an amount of 0.001% of the total drug weight.

[0030] Carbonic acid amide in an amount of 0.01% of the total drug weight was added to a mixture of a strongly solvating aprotic solvent from the group of sulfoxides in an amount of 0.01% of the total drug weight, vegetable oil D with a high content of unsaturated fatty acids in an amount of 1% of the total drug weight, and optionally a natural emulsifying substance in an amount of 0.001% of the total drug weight.

[0031] The resulting mixture was stirred until a homogenous consistency and color was achieved.

[0032] The resulting mixture exhibited emulsifying properties. Hydrophilic substances were dissolved in the molten mixture. Lipophilic substances were dissolved in the mixture at room temperature. Example 3

[0033] The carrier mixture (F1) was prepared in the following steps:

[0034] a) A vegetable oil having a high content of unsaturated fatty acids in an amount of 5% of the total weight of the drug, and optionally a natural emulsifying agent in an amount of 8% of the total weight of the drug, were melted in a vessel at a maximum of 40°C.

[0035] b) A strongly solvating aprotic solvent from the group of sulfoxides in an amount of 5% of the total drug weight was added under stirring to a molten mixture of a vegetable oil with a high content of unsaturated fatty acids in an amount of 5% of the total drug weight and optionally a natural emulsifying substance in an amount of 8% of the total drug weight.

[0036] Carbonic acid amide in an amount of 2% of the total drug weight was added to a mixture of a strongly solvating aprotic solvent from the group of sulfoxides in an amount of 5% of the total drug weight, vegetable oil D with a high content of unsaturated fatty acids in an amount of 5% of the total drug weight, and optionally a natural emulsifying substance in an amount of 8% of the total drug weight.

[0037] The resulting mixture was stirred until a homogenous consistency and color was achieved.

[0038] The resulting mixture also exhibited emulsifying properties. Hydrophilic substances were dissolved in the molten mixture. Lipophilic substances were dissolved in the mixture at room temperature.

[0039] As a result of carrying out Examples 1, 2 and 3, mixtures were obtained within the quantitative (percentage) ranges of the individual components used in the carrier, proving that the assumption of the first object of the present invention was met.

[0040] The second object of the present invention is the use of a unique mixture of the above-mentioned carriers in semi-solid drug form and active substances in appropriately selected ratios. The active substances are mixtures of all synthetic and natural origin substances, such as Coptis Chinensis and Curcuma Longa extracts. The weight ratio between the active substances and the carrier should be in the range of 1:10 to 2:1. The percentages given are the concentrations in 100 parts of the final product. The carriers constitute 1.0 to 20% by weight of the final product.

[0041] The main component of the substrate is a mixture obtained from lard in an amount of 5-20%, goose fat in an amount of 4-15%, eucerin in an amount of 12-21% and zinc sulfate in an amount of 0.5-2%, the weight ratio of active substance to excipients is 10:90-1:999, and has the form of a semi-solid drug formulation, characterized in that it contains:

[0042] a) Mixing the lipophilic substance and a portion of the extract, preferably in an amount of 0.1%-5%, with the substrate required for the entire process, in a pharmaceutical and food mixer at a temperature range of 20-50°C, at a maximum of 4000 RPM, for at least 3 minutes to achieve powderization of the extract.

[0043] b) Mixing the resulting mixture from step a) with a portion, preferably 6%-90%, of the substrate required for the entire process, remaining amount, in a pharmaceutical and food mixer at a temperature range of 20-50° C., at maximum 5000 RPM, for at least 3 minutes.

[0044] c) Mixing the resulting mixture from step b) with the remaining amount of substrate required for the entire process, and then with the remaining excipients, in a pharmaceutical and food grade mixer at a maximum of 4000 RPM for at least 3 minutes to achieve homogeneity, at a temperature range of 20-50°C.

[0045] d) Homogenization in laboratory and industrial (rotary and piston) homogenizers at temperatures ranging from 20 to 50°C for at least 5 minutes.

[0046] The essence of the present invention is to create a homogeneous mixture of substrate and active substance, containing the active substance evenly distributed. According to the present invention, a semi-solid mixture was obtained, which was dispensed into pharmaceutical packaging for ointments. Example 4

[0047] Process for obtaining the ointment according to the invention The products that are the object of the present invention, the preferred embodiments of which are shown in Table 3, were prepared by the following manufacturing techniques:

[0048] An appropriately weighed amount of plant extract was mixed with 9% of the required amount of carrier in a rotary homogenizer (1000 RPM for 7 minutes), then 5% of the required amount of molten substrate was added to the homogenizer and the entire mixture was pulverized in the rotary homogenizer (500 RPM for 7 minutes). The remaining amount of carrier was added to the rotary homogenizer and mixed, then the remainder of the molten substrate was added and homogenized (2000 RPM for 10 minutes).

[0049] The resulting ointment was then transferred to pharmaceutical packaging for ointments.

[0050] [Table 3]

[0051] A semi-solid mixture is obtained which is homogeneous with respect to the content of active substances, and the content of impurities in the resulting plant extract is below the detection level due to the experimentally selected pharmaceutical composition, the appropriate subdivision of the components of this composition and the manufacturing technology used.

[0052] The selection of suitable compositions in experimentally selected quantitative proportions and the use of natural medicinal substances, specific excipients and the application of the above-mentioned ointment mixture manufacturing technology make it possible to obtain a uniform content of active substance in the form of a drug, i.e. the same dose of active substance is present in each volume unit, confirmation of which was obtained during testing.

[0053] The test was performed on L929 cell line (mouse fibroblasts) from Sigma Aldrich, catalog number 85011425, item number 14G010. Cells from cryotube no. 94 were used for the test. The given cell line was selected for the test based on the PN-ISO 10993-5 standard.

[0054] Cytotoxicity tests using the XTT method were carried out according to PN-ISO 10993-5 based on the "Test Instruction-XTT" (IB 16.4).

[0055] result *Limit of viability - 70%

[0056] [Table 4]

[0057] [Table 5]

[0058] [Table 6]

[0059] [Table 7]

[0060] [Table 8]

[0061] overview: The average % viability for all five samples was greater than 70%*, the results indicating that the test material was not cytotoxic.

[0062] In addition, the use of carriers ensures protection of the active substances against the adverse effects of external environmental conditions, i.e. resistance to temperature and light during technological processing. Example 5

[0063] Method for obtaining the ointment according to the invention The products that are the object of the present invention, the preferred embodiments of which are shown in Table 3, were prepared by the following manufacturing techniques:

[0064] A properly weighed amount of plant extract was mixed with 100% of the required amount of carrier in a rotary homogenizer (1000RPM for 7 minutes at room temperature) and then heated in a water bath until Mix-1 was liquefied. Then 25% of the required amount of substrate was added to the liquefied mixture and then mixed until a homogenous consistency (Mix-2) was obtained. The remaining amount of substrate was added to the rotary homogenizer and mixed with the liquefied Mix-2 (500RPM for 7 minutes at room temperature) and then the entire mixture was homogenized (2000RPM for 10 minutes).

[0065] The resulting ointment mixture was then transferred into pharmaceutical packaging for ointments.

[0066] [Table 9]

[0067] A semi-solid mixture was obtained which was homogeneous with respect to the content of active substance, the content of impurities of the resulting plant extract being below the detection level due to the experimentally selected pharmaceutical composition, the appropriate subdivision of the components of this composition and the manufacturing technology used, which was confirmed by tests allowing the determination of the following product parameters: Iodine Value (A) Method (Ph.Eur.): Titration Iodine value: 41.20

[0068] The iodine value is the number of grams of elements derived from halogen radicals, calculated as iodine, attached to a specific double bond present in the fatty acids found in 100 grams of the tested product. The iodine value can be used to identify and parameterize products in order to determine the level of unsaturation of the fat. Products can differ within certain limits in the composition of fatty acids, and therefore in the iodine value as well, depending on the origin of the fat raw material. Apparent Viscosity (A) Method (Ph.Eur.): Viscosity measurement Apparent viscosity: 836.7mPa×s

[0069] The results of the tests will be necessary to determine the product standard. At this level of viscosity it is possible to maintain the active substance in the form of a homogeneous mixture.

[0070] The selection of suitable compositions in experimentally selected quantitative proportions, as well as the use of natural medicinal substances, specific excipients and the application of the above-mentioned ointment mixture manufacturing technology, makes it possible to obtain a uniform content of active substances in the form of a drug.

[0071] In addition, the use of a carrier ensures protection of the active substance against the harmful effects of external environmental conditions during technological processing. Acid Number (A) Method (Ph.Eur.): Titration Acid value 0.12

[0072] The test carried out (acid number) determined the content of free fatty acids (degree of fat hydrolysis) in the tested products. Saponification value (A) Method (Ph.Eur.): Titration Saponification value: 96.2

[0073] The test determined the average molecular weight of the fatty acids, and the result was the number of milligrams of potassium hydroxide required to neutralize the free fatty acids and saponify the acylglycerols contained in 1 gram of the test product. Dropping point Method (Ph.Eur.): Visual evaluation Melting point: 24.0℃±2%

[0074] The results of the testing will be necessary to determine product standards. Peroxide Value (USP)(A) Method (USP): Titration Peroxide value 0.20

[0075] The peroxide value is the content of peroxides and is taken as an index of the degree of oxidation (rancidity) of fats. The values ​​obtained for the products are within the normal range. Example 6

[0076] Method for the preparation of a pharmaceutical product containing lipophilic and hydrophilic substances Equivalent amounts of substances from the group of vitamins were used as reference substances: a) Vitamin A - fat-soluble substance b) Vitamin C - Water-soluble substance

[0077] [Table 10]

[0078] The composition ratio of the F1 carrier described in Example 1 was used in the experiments. The following amounts of active substance: - Vitamin A - 1g - Vitamin C - 1g was added to a carrier composition in the amount of 20 g obtained according to the procedure described for F1 and melted at 40 degrees Celsius.

[0079] After mixing (200 RPM) for about 10 minutes, a homogenous mixture was obtained and the substances used were completely dissolved after mixing. The mixture so obtained was added to the substrate (80 g) described in Example 4 with stirring (500 RPM) for 8 minutes.

[0080] As a result, a formulation of homogenous consistency and color was obtained with a uniformly powdered substance, the ratio between active substance and carrier was 1:10 and the ratio between carrier and substrate was 1:5. Example 7

[0081] Method for the preparation of a pharmaceutical product containing lipophilic and hydrophilic substances Equivalent amounts of substances from the group of vitamins were used as reference substances: a) Vitamin A - fat-soluble substance b) Vitamin C - Water-soluble substance

[0082] [Table 11]

[0083] The composition ratio of the F1 carrier described in Example 1 was used in the experiments. The following amounts of active substance: - Vitamin A - 0.05g - Vitamin C - 0.05g was added to a 1 g amount of carrier composition obtained according to the procedure described for F1 and melted at 40 degrees Celsius.

[0084] After mixing (200 RPM) for about 10 minutes, a homogenous mixture was obtained and the substances used were completely dissolved after mixing. The mixture thus obtained was added to the substrate (98.8 g) described in Example 4 with stirring (500 RPM) for 8 minutes.

[0085] As a result, a formulation of homogenous consistency and color was obtained with a uniformly powdered substance, the ratio between active substance and carrier being 1:10 and the ratio between carrier and substrate being 1:99. Example 8

[0086] Method for the preparation of a pharmaceutical product containing lipophilic and hydrophilic substances Equivalent amounts of substances from the group of vitamins were used as reference substances: a) Vitamin A - fat-soluble substance b) Vitamin C - Water-soluble substance

[0087] [Table 12]

[0088] The composition ratio of the F1 carrier described in Example 1 was used in the experiments. The following amounts of active substance: - Vitamin A - 20g - Vitamin C - 20g was added to a carrier composition in the amount of 20 g obtained according to the procedure described for F1 and melted at 40 degrees Celsius.

[0089] After mixing (400 RPM) for about 10 minutes, a homogenous mixture was obtained and the substances used were completely dissolved after mixing. The mixture so obtained was added to the remaining amount of substrate (40 g) described in Example 4 with stirring (500 RPM) for 8 minutes.

[0090] As a result, a formulation of homogenous consistency and color was obtained with a uniformly powdered substance, the ratio between active substance and carrier was 2:1, the ratio between carrier and substrate was 1:2, and the amount of carrier constituted 20% of the total weight of the final product. Example 9

[0091] Method for the preparation of a pharmaceutical product containing lipophilic and hydrophilic substances Equivalent amounts of substances from the group of vitamins were used as reference substances: a) Vitamin A - fat-soluble substance b) Vitamin C - Water-soluble substance

[0092] [Table 13]

[0093] The composition ratio of the F1 carrier described in Example 1 was used in the experiments. The following amounts of active substance: - Vitamin A - 1g - Vitamin C - 1g was added to a 1 g amount of carrier composition obtained according to the procedure described for F1 and melted at 40 degrees Celsius.

[0094] After mixing (400 RPM) for about 10 minutes, a homogenous mixture was obtained and the materials used were completely dissolved after mixing. The mixture so obtained was added to the remaining amount of substrate (97 g) described in Example 4 with stirring (500 RPM) for 8 minutes.

[0095] As a result, a formulation of homogenous consistency and color was obtained with a uniformly powdered substance, the ratio between active substance and carrier was 2:1, the ratio between carrier and substrate was 1:2, and the amount of carrier constituted 1% of the total weight of the final product. Example 10

[0096] Method for the preparation of a pharmaceutical product containing lipophilic and hydrophilic substances Equivalent amounts of substances from the group of vitamins were used as reference substances: a) Vitamin A - fat-soluble substance b) Vitamin C - Water-soluble substance

[0097] [Table 14]

[0098] The composition ratio of the F1 carrier described in Example 1 was used in the experiments. The following amounts of active substance: - Vitamin A - 1g - Vitamin C - 1g was added to a carrier composition in the amount of 20 g obtained according to the procedure described for F1 and melted at 40 degrees Celsius.

[0099] After mixing (200 RPM) for about 10 minutes, a homogenous mixture was obtained and the substances used were completely dissolved after mixing. The mixture so obtained was added to the substrate (80 g) described in Example 4 (with one important difference: a similar amount of Lekobaza was used instead of Eucerin) with stirring (500 RPM) for 8 minutes.

[0100] As a result, a formulation of homogenous consistency and color was obtained with a uniformly powdered substance, the ratio between active substance and carrier was 1:10 and the ratio between carrier and substrate was 1:5. Example 11

[0101] Method for the preparation of a pharmaceutical product containing lipophilic and hydrophilic substances Equivalent amounts of substances from the group of vitamins were used as reference substances: a) Vitamin A - fat-soluble substance b) Vitamin C - Water-soluble substance

[0102] [Table 15]

[0103] The composition ratio of the F1 carrier described in Example 1 was used in the experiments. The following amounts of active substance: - Vitamin A - 1g - Vitamin C - 1g was added to a carrier composition in the amount of 20 g obtained according to the procedure described for F1 and melted at 40 degrees Celsius.

[0104] After mixing (200 RPM) for about 10 minutes, a homogenous mixture was obtained and the substances used were completely dissolved after mixing. The mixture so obtained was added to the substrate (80 g) described in Example 4 (with one important difference: a similar amount of Hascobaza was used instead of Eucerin) with stirring (500 RPM) for 8 minutes.

[0105] As a result, a formulation of homogenous consistency and color was obtained with a uniformly powdered substance, the ratio between active substance and carrier was 1:10 and the ratio between carrier and substrate was 1:5. Example 12

[0106] Method for the preparation of a pharmaceutical product containing lipophilic and hydrophilic substances Equivalent amounts of substances from the group of vitamins were used as reference substances: a) Vitamin A - fat-soluble substance b) Vitamin C - Water-soluble substance

[0107] [Table 16]

[0108] The composition ratio of the F1 carrier described in Example 1 was used in the experiments. The following amounts of active substance: - Vitamin A - 1g - Vitamin C - 1g was added to a carrier composition in the amount of 20 g obtained according to the procedure described for F1 and melted at 40 degrees Celsius.

[0109] After mixing (200 RPM) for about 10 minutes, a homogenous mixture was obtained and the substances used were completely dissolved after mixing. The mixture so obtained was added to the substrate (80 g) described in Example 4 (with one important difference: a similar amount of yellow petrolatum was used instead of Eucerin) with stirring (500 RPM) for 8 minutes.

[0110] As a result, a formulation of homogenous consistency and color was obtained with a uniformly powdered substance, the ratio between active substance and carrier was 1:10 and the ratio between carrier and substrate was 1:5. Example 13

[0111] Method for the preparation of a pharmaceutical product containing lipophilic and hydrophilic substances Equivalent amounts of substances from the group of vitamins were used as reference substances: a) Vitamin A - fat-soluble substance b) Vitamin C - Water-soluble substance

[0112] [Table 17]

[0113] The composition ratio of the F1 carrier described in Example 1 was used in the experiments. The following amounts of active substance: - Vitamin A - 1g - Vitamin C - 1g was added to a carrier composition in the amount of 20 g obtained according to the procedure described for F1 and melted at 40 degrees Celsius.

[0114] After mixing (200 RPM) for about 10 minutes, a homogenous mixture was obtained and the substances used were completely dissolved after mixing. The mixture so obtained was added to the substrate (80 g) described in Example 4 (with one important difference: a similar amount of white petrolatum was used instead of Eucerin) with stirring (500 RPM) for 8 minutes.

[0115] As a result, a formulation of homogenous consistency and color was obtained with a uniformly powdered substance, the ratio between active substance and carrier was 1:10 and the ratio between carrier and substrate was 1:5. Example 14

[0116] Method for the preparation of a pharmaceutical product containing lipophilic and hydrophilic substances Equivalent amounts of substances from the group of vitamins were used as reference substances: a) Vitamin A - fat-soluble substance b) Vitamin C - Water-soluble substance

[0117] [Table 18]

[0118] The composition ratio of the F1 carrier described in Example 1 was used in the experiments. The following amounts of active substance: - Vitamin A - 1g - Vitamin C - 1g was added to a carrier composition in the amount of 20 g obtained according to the procedure described for F1 and melted at 40 degrees Celsius.

[0119] After mixing (200 RPM) for about 10 minutes, a homogeneous mixture was obtained and the substances used were completely dissolved after mixing. The mixture so obtained was added to the substrate (80 g) described in Example 4 (with one important difference: a similar amount of Lekobaza was used instead of Eucerin and lard was used instead of goose fat) with stirring (500 RPM) for 8 minutes.

[0120] As a result, a formulation of homogenous consistency and color was obtained with a uniformly powdered substance, the ratio between active substance and carrier was 1:10 and the ratio between carrier and substrate was 1:5. Example 15

[0121] Method for the preparation of a pharmaceutical product containing lipophilic and hydrophilic substances Equivalent amounts of substances from the group of vitamins were used as reference substances: a) Vitamin A - fat-soluble substance b) Vitamin C - Water-soluble substance

[0122] [Table 19]

[0123] The composition ratio of the F1 carrier described in Example 1 was used in the experiments. The following amounts of active substance: - Vitamin A - 1g - Vitamin C - 1g was added to a carrier composition in the amount of 20 g obtained according to the procedure described for F1 and melted at 40 degrees Celsius.

[0124] After mixing (200 RPM) for about 10 minutes, a homogenous mixture was obtained and the substances used were completely dissolved after mixing. The mixture so obtained was added to the substrate (80 g) described in Example 4 (with one important difference: a similar amount of Hascobaza was used instead of Eucerin and duck fat was used instead of goose fat) with stirring (500 RPM) for 8 minutes.

[0125] As a result, a formulation of homogenous consistency and color was obtained with a uniformly powdered substance, the ratio between active substance and carrier was 1:10 and the ratio between carrier and substrate was 1:5. Example 16

[0126] Method for the preparation of a pharmaceutical product containing lipophilic and hydrophilic substances Equivalent amounts of substances from the group of vitamins were used as reference substances: a) Vitamin A - fat-soluble substance b) Vitamin C - Water-soluble substance

[0127] [Table 20]

[0128] The composition ratio of the F1 carrier described in Example 1 was used in the experiments. The following amounts of active substance: - Vitamin A - 1g - Vitamin C - 1g was added to a carrier composition in the amount of 20 g obtained according to the procedure described for F1 and melted at 40 degrees Celsius.

[0129] After about 10 minutes of mixing (200 RPM), a homogeneous mixture was obtained and the substances used were completely dissolved after mixing. The mixture thus obtained was added to the substrate (80 g) described in Example 4 (with one important difference: a similar amount of white petrolatum was used instead of Eucerin, lard was used instead of goose fat, and a similar amount of zinc chloride was used instead of zinc sulfate) with stirring (500 RPM) for 8 minutes.

[0130] As a result, a formulation of homogenous consistency and color was obtained with a uniformly powdered substance, the ratio between active substance and carrier was 1:10 and the ratio between carrier and substrate was 1:5. Example 17

[0131] Method for the preparation of a pharmaceutical product containing lipophilic and hydrophilic substances Equivalent amounts of substances from the group of vitamins were used as reference substances: a) Vitamin A - fat-soluble substance b) Vitamin C - Water-soluble substance

[0132] [Table 21]

[0133] The composition ratio of the F1 carrier described in Example 1 was used in the experiments. The following amounts of active substance: - Vitamin A - 1g - Vitamin C - 1g was added to a 20 g amount of carrier composition obtained according to the procedure described for F1 (with one important difference: a similar amount of linseed oil was used instead of linseed oil) and melted at 40 degrees Celsius.

[0134] After mixing (200 RPM) for about 10 minutes, a homogenous mixture was obtained and the substances used were completely dissolved after mixing. The mixture so obtained was added to the substrate (80 g) described in Example 4 with stirring (500 RPM) for 8 minutes.

[0135] As a result, a formulation of homogenous consistency and color was obtained with a uniformly powdered substance, the ratio between active substance and carrier was 1:10 and the ratio between carrier and substrate was 1:5. Example 18

[0136] Method for the preparation of a pharmaceutical product containing lipophilic and hydrophilic substances Equivalent amounts of substances from the group of vitamins were used as reference substances: a) Vitamin A - fat-soluble substance b) Vitamin C - Water-soluble substance

[0137] [Table 22]

[0138] The composition ratio of the F1 carrier described in Example 1 was used in the experiments. The following amounts of active substance: - Vitamin A - 1g - Vitamin C - 1g was added to a 20 g amount of carrier composition obtained according to the procedure described for F1 (with one important difference: a similar amount of rapeseed oil was used instead of canola oil) and melted at 40 degrees Celsius.

[0139] After mixing (200 RPM) for about 10 minutes, a homogenous mixture was obtained and the substances used were completely dissolved after mixing. The mixture so obtained was added to the substrate (80 g) described in Example 4 with stirring (500 RPM) for 8 minutes.

[0140] As a result, a formulation of homogenous consistency and color was obtained with a uniformly powdered substance, the ratio between active substance and carrier was 1:10 and the ratio between carrier and substrate was 1:5. Example 19

[0141] Method for the preparation of a pharmaceutical product containing lipophilic and hydrophilic substances Equivalent amounts of substances from the group of vitamins were used as reference substances: a) Vitamin A - fat-soluble substance b) Vitamin C - Water-soluble substance

[0142] [Table 23]

[0143] The composition ratio of the F1 carrier described in Example 1 was used in the experiments. The following amounts of active substance: - Vitamin A - 1g - Vitamin C - 1g was added to a 20 g amount of carrier composition obtained according to the procedure described for F1 (with one important difference: a similar amount of peanut oil was used instead of peanut oil) and melted at 40 degrees Celsius.

[0144] After mixing (200 RPM) for about 10 minutes, a homogenous mixture was obtained and the substances used were completely dissolved after mixing. The mixture so obtained was added to the substrate (80 g) described in Example 4 with stirring (500 RPM) for 8 minutes.

[0145] As a result, a formulation of homogenous consistency and color was obtained with a uniformly powdered substance, the ratio between active substance and carrier was 1:10 and the ratio between carrier and substrate was 1:5. Example 20

[0146] Method for the preparation of a pharmaceutical product containing lipophilic and hydrophilic substances Equivalent amounts of substances from the group of vitamins were used as reference substances: a) Vitamin A - fat-soluble substance b) Vitamin C - Water-soluble substance

[0147] [Table 24]

[0148] The composition ratio of the F1 carrier described in Example 1 was used in the experiments. The following amounts of active substance: - Vitamin A - 1g - Vitamin C - 1g was added to a 20 g amount of carrier composition obtained according to the procedure described for F1 (with one important difference: a similar amount of sunflower oil was used instead of douche oil) and melted at 40 degrees Celsius.

[0149] After mixing (200 RPM) for about 10 minutes, a homogenous mixture was obtained and the substances used were completely dissolved after mixing. The mixture thus obtained was added to the substrate (80 g) described in Example 4 with stirring (500 RPM) for 8 minutes.

[0150] As a result, a formulation of homogenous consistency and color was obtained with a uniformly powdered substance, the ratio between active substance and carrier was 1:10 and the ratio between carrier and substrate was 1:5. Example 21

[0151] Method for the preparation of a pharmaceutical product containing lipophilic and hydrophilic substances Equivalent amounts of substances from the group of vitamins were used as reference substances: a) Vitamin A - fat-soluble substance b) Vitamin C - Water-soluble substance

[0152] [Table 25]

[0153] The composition ratio of the F1 carrier described in Example 1 was used in the experiments. The following amounts of active substance: - Vitamin A - 1g - Vitamin C - 1g was added to a 20 g amount of carrier composition obtained according to the procedure described for F1 (with one important difference: a similar amount of hemp seed oil was used instead of douche oil) and melted at 40 degrees Celsius.

[0154] After mixing (200 RPM) for about 10 minutes, a homogenous mixture was obtained and the substances used were completely dissolved after mixing. The mixture thus obtained was added to the substrate (80 g) described in Example 4 with stirring (500 RPM) for 8 minutes.

[0155] As a result, a formulation of homogenous consistency and color was obtained with a uniformly powdered substance, the ratio between active substance and carrier was 1:10 and the ratio between carrier and substrate was 1:5. Example 22

[0156] Method for the preparation of medicinal products containing substances from the group of sodium-glucose cotransporter 2 inhibitors (SGLT2) Substances from the group of sodium-glucose cotransporter 2 (SGLT2) inhibitors were used as reference substances:

[0157] Dapagliflozin propanediol Dapagliflozin is a highly potent (Ki: 0.55 nM), selective and reversible inhibitor of sodium-glucose cotransporter 2 (SGLT2).

[0158] SGLT2 is selectively expressed in the kidney with a concomitant lack of expression in over 70 other types of tissue, including the liver, muscle tissue, adipose tissue, breast, bladder and brain. SGLT2 is the main transporter involved in glucose reabsorption from glomerular filtration into the blood. In type 2 diabetes, glucose reabsorption still occurs in the kidney despite the presence of hyperglycemia. Dapagliflozin improves fasting and postprandial glucose control by decreasing renal glucose reabsorption and resulting in urinary excretion of glucose. Glucose excretion is observed after the first dose, continues during the 24-hour interval between doses, and persists throughout the treatment period. The amount of glucose removed by the kidney due to these mechanisms of action depends on the blood glucose level and the degree of glomerular filtration rate (GFR). Dapagliflozin does not interfere with normal endogenous glucose production in response to hypoglycemia. Dapagliflozin functions independently of insulin secretion and action. Improvements in HOMA beta cells (homeostasis model assessment of beta cell function) were observed in preclinical studies.

[0159] Dapagliflozin-induced urinary excretion of glucose is associated with calorie consumption and weight loss. Inhibition of glucose and sodium cotransporters by dapagliflozin is also associated with mild diuresis and transient natriuresis.

[0160] Dapagliflozin does not inhibit other glucose transporters important for glucose transport to peripheral tissues and is >1400-fold more selective for SGLT2 than SGLT1, the main transporter responsible for glucose absorption in the intestine.

[0161] The composition ratio of the F1 carrier described in Example 1 was used in the experiment. An amount of 5 mg of active substance was added to an amount of 20 g of the carrier composition obtained according to the procedure described for F1 and melted at 40 degrees Celsius.

[0162] After mixing (200 RPM) for about 10 minutes, a homogenous mixture was obtained and the substances used were completely dissolved after mixing. The mixture thus obtained was added to the substrate (80 g) described in Example 4 with stirring (500 RPM) for 8 minutes.

[0163] As a result, a formulation of homogenous consistency and color was obtained with a uniformly powdered substance, the ratio between active substance and carrier was 1:10 and the ratio between carrier and substrate was 1:5. Example 23

[0164] Method for the preparation of medicinal products containing substances from the group of sodium-glucose cotransporter 2 inhibitors (SGLT2) Substances from the group of sodium glucose cotransporter 2 (SGLT2) inhibitors were used as reference substances:

[0165] Dapagliflozin propanediol The composition ratio of the F1 carrier described in Example 1 was used in the experiment. An amount of 10 mg of active substance was added to an amount of 1 g of carrier composition obtained according to the procedure described for F1 and melted at 40 degrees Celsius.

[0166] After mixing (200 RPM) for about 10 minutes, a homogenous mixture was obtained and the substances used were completely dissolved after mixing. The mixture thus obtained was added to the substrate (98.8 g) described in Example 4 with stirring (500 RPM) for 8 minutes.

[0167] As a result, a formulation of homogenous consistency and color was obtained with a uniformly powdered material and a ratio between carrier and substrate of 1:99. Example 24

[0168] Method for the preparation of medicinal products containing substances from the group of sodium-glucose cotransporter 2 inhibitors (SGLT2) Substances from the group of sodium-glucose cotransporter 2 (SGLT2) inhibitors were used as reference substances:

[0169] Dapagliflozin propanediol The composition ratio of F1 carrier described in Example 1 was used in the experiment. A portion of the carrier described in Example 4 in an amount of 20 g was added to a carrier composition obtained according to the procedure described for F1 and melted at 40 degrees Celsius in an amount of 20 g, followed by an active substance in an amount of 20 mg.

[0170] After mixing (400 RPM) for about 10 minutes, a homogenous mixture was obtained and the substances used were completely dissolved after mixing. The mixture so obtained was added to the remaining substrate (20 g) described in Example 4 with stirring (500 RPM) for 8 minutes.

[0171] As a result, a formulation of homogenous consistency and color was obtained with a uniformly powdered substance, the ratio between carrier and substrate was 1:2, and the amount of carrier constituted 20% of the total weight of the final product. Example 25

[0172] Method for the preparation of medicinal products containing substances from the group of sodium-glucose cotransporter 2 inhibitors (SGLT2) Substances from the group of sodium-glucose cotransporter 2 (SGLT2) inhibitors were used as reference substances:

[0173] Dapagliflozin propanediol The composition ratio of F1 carrier described in Example 1 was used in the experiment. A portion of the carrier described in Example 4 in an amount of 1 g was added to a carrier composition obtained according to the procedure described for F1 and melted at 40 degrees Celsius in an amount of 1 g, followed by an active substance in an amount of 35 mg.

[0174] After mixing (400 RPM) for about 10 minutes, a homogenous mixture was obtained and the materials used were completely dissolved after mixing. The mixture so obtained was added to the remaining substrate (96 g) described in Example 4 with stirring (500 RPM) for 8 minutes.

[0175] As a result, a formulation of homogenous consistency and color was obtained with a uniformly powdered substance, the ratio between carrier and substrate was 1:97, and the amount of carrier constituted 1% of the total weight of the final product. Example 26

[0176] Method for the preparation of medicinal products containing substances from the group of sodium-glucose cotransporter 2 inhibitors (SGLT2) Substances from the group of sodium-glucose cotransporter 2 (SGLT2) inhibitors were used as reference substances:

[0177] Dapagliflozin propanediol The composition ratio of the F1 carrier described in Example 1 was used in the experiment. An amount of 50 mg of active substance was added to an amount of 20 g of the carrier composition obtained according to the procedure described for F1 and melted at 40 degrees Celsius.

[0178] After mixing (200 RPM) for about 10 minutes, a homogenous mixture was obtained and the substances used were completely dissolved after mixing. The mixture so obtained was added to the substrate (80 g) described in Example 4 (with one important difference: a similar amount of Lekobaza was used instead of Eucerin) with stirring (500 RPM) for 8 minutes.

[0179] As a result, a formulation of homogenous consistency and color was obtained with a uniformly powdered material and a ratio between carrier and substrate of 1:5. Example 27

[0180] Method for the preparation of medicinal products containing substances from the group of sodium-glucose cotransporter 2 inhibitors (SGLT2) Substances from the group of sodium-glucose cotransporter 2 (SGLT2) inhibitors were used as reference substances:

[0181] Dapagliflozin propanediol The composition ratio of the F1 carrier described in Example 1 was used in the experiment. An amount of 70 mg of active substance was added to an amount of 20 g of carrier composition obtained according to the procedure described for F1 and melted at 40 degrees Celsius.

[0182] After mixing (200 RPM) for about 10 minutes, a homogenous mixture was obtained and the substances used were completely dissolved after mixing. The mixture so obtained was added to the substrate (80 g) described in Example 4 (with one important difference: a similar amount of Hascobaza was used instead of Eucerin) with stirring (500 RPM) for 8 minutes.

[0183] As a result, a formulation of homogenous consistency and color was obtained with a uniformly powdered material and a ratio between carrier and substrate of 1:5. Example 28

[0184] Method for the preparation of medicinal products containing substances from the group of sodium-glucose cotransporter 2 inhibitors (SGLT2) Substances from the group of sodium-glucose cotransporter 2 (SGLT2) inhibitors were used as reference substances:

[0185] Dapagliflozin propanediol The composition ratio of the F1 carrier described in Example 1 was used in the experiment. An amount of 100 mg of active substance was added to an amount of 20 g of the carrier composition obtained according to the procedure described for F1 and melted at 40 degrees Celsius.

[0186] After mixing (200 RPM) for about 10 minutes, a homogenous mixture was obtained and the substances used were completely dissolved after mixing. The mixture so obtained was added to the substrate (80 g) described in Example 4 (with one important difference: a similar amount of yellow petrolatum was used instead of Eucerin) with stirring (500 RPM) for 8 minutes.

[0187] As a result, a formulation of homogenous consistency and color was obtained with a uniformly powdered material and a ratio between carrier and substrate of 1:5. Example 29

[0188] Method for the preparation of medicinal products containing substances from the group of sodium-glucose cotransporter 2 inhibitors (SGLT2) Substances from the group of sodium-glucose cotransporter 2 (SGLT2) inhibitors were used as reference substances:

[0189] Dapagliflozin propanediol The composition ratio of the F1 carrier described in Example 1 was used in the experiment. An amount of 105 mg of active substance was added to an amount of 20 g of the carrier composition obtained according to the procedure described for F1 and melted at 40 degrees Celsius.

[0190] After mixing (200 RPM) for about 10 minutes, a homogenous mixture was obtained and the substances used were completely dissolved after mixing. The mixture so obtained was added to the substrate (80 g) described in Example 4 (with one important difference: a similar amount of white petrolatum was used instead of Eucerin) with stirring (500 RPM) for 8 minutes.

[0191] As a result, a formulation of homogenous consistency and color was obtained with a uniformly powdered material and a ratio between carrier and substrate of 1:5.

[0192] The composition ratio of the F1 carrier described in Example 1 was used in the experiment: an amount of 140 mg of active substance was added to an amount of 20 g of carrier composition obtained according to the procedure described for F1 and melted at 40 degrees Celsius.

[0193] After mixing (200 RPM) for about 10 minutes, a homogeneous mixture was obtained and the substances used were completely dissolved after mixing. The mixture so obtained was added to the substrate (80 g) described in Example 4 (with one important difference: a similar amount of Lekobaza was used instead of Eucerin and lard was used instead of goose fat) with stirring (500 RPM) for 8 minutes.

[0194] As a result, a formulation of homogenous consistency and color was obtained with a uniformly powdered material and a ratio between carrier and substrate of 1:5. Example 30

[0195] Method for the preparation of medicinal products containing substances from the group of sodium-glucose cotransporter 2 inhibitors (SGLT2) Substances from the group of sodium-glucose cotransporter 2 (SGLT2) inhibitors were used as reference substances:

[0196] Dapagliflozin propanediol The composition ratio of the F1 carrier described in Example 1 was used in the experiment. An amount of 210 mg of active substance was added to an amount of 20 g of carrier composition obtained according to the procedure described for F1 and melted at 40 degrees Celsius.

[0197] After mixing (200 RPM) for about 10 minutes, a homogenous mixture was obtained and the materials used were completely dissolved after mixing. The mixture so obtained was added to the substrate (80 g) described in Example 4 (with one important difference: a similar amount of white Hascobaza was used instead of Eucerin and duck fat was used instead of goose fat) with stirring (500 RPM) for 8 minutes.

[0198] As a result, a formulation of homogenous consistency and color was obtained with a uniformly powdered material and a ratio between carrier and substrate of 1:5. Example 31

[0199] Method for the preparation of medicinal products containing substances from the group of sodium-glucose cotransporter 2 inhibitors (SGLT2) Substances from the group of sodium-glucose cotransporter 2 (SGLT2) inhibitors were used as reference substances:

[0200] Dapagliflozin propanediol The composition ratio of the F1 carrier described in Example 1 was used in the experiment. An amount of 240 mg of active substance was added to an amount of 20 g of carrier composition obtained according to the procedure described for F1 and melted at 40 degrees Celsius.

[0201] After about 10 minutes of mixing (200 RPM), a homogeneous mixture was obtained and the substances used were completely dissolved after mixing. The mixture thus obtained was added to the substrate (80 g) described in Example 4 (with one important difference: a similar amount of white petrolatum was used instead of Eucerin, lard was used instead of goose fat, and a similar amount of zinc chloride was used instead of zinc sulfate) with stirring (500 RPM) for 8 minutes.

[0202] As a result, a formulation of homogenous consistency and color was obtained with a uniformly powdered material and a ratio between carrier and substrate of 1:5.

[0203] To determine the possibility of transporting substances of different physicochemical properties through the skin and their effect on the bioavailability, studies were carried out using fluorescent markers (fluorescein, rhodamine, acridine orange) on animal models.

[0204] Fluorescein (C20H12O5) - a xanthene dye, an organic chemical compound with a molecular weight of 332.31 g / mol, a xanthene derivative. In alkaline solutions, fluorescein exhibits green-yellow fluorescence that is visible even at dilutions of 1 in tens of millions.

[0205] Acridine orange (C17H19N3) is an organic compound with a molecular weight of 265.36 g / mol. It is used as a nucleic acid selective cationic fluorescent dye and is useful for determining the cell cycle. It is membrane permeable and therefore interacts with DNA and RNA by intercalation or electrostatic interactions, respectively.

[0206] Rhodamine B (C28H31N2O3Cl) - an organic chemical compound derived from the rhodamine group with a molecular weight of 479.01 g / mol. It is used as a fluorescent dye, i.e. to stain biological products.

[0207] Test materials were first equilibrated at 4°C for 1 hour and then frozen. Materials were cut at -20°C in a Leica CM1950 cryostat. 10 μm thick sections were fixed in 70% alcohol and mounted in Euperal. Materials were analyzed using a Nikon Eclipse80i fluorescent microscope equipped with UV-2A (EX450-490, DM-500, BA-515) and B-2A ((EX330-380, DM-400, BA-420) filters. Histological preparations, routinely stained with hematoxylin and eosin, were made from the sections. All images were taken with a constant exposure time of 400 ms. Selected images were then corrected with Nis elements Ar software to reveal structures.

[0208] The study was performed on Buffalo rats, randomized and housed separately in cages with toys under standard conditions in the animal facility of the Department of Biostructure and Animal Physiology.

[0209] The animals were divided into 12 groups of 3 animals each.

[0210] The skin on the back near the terminal segment of the ribs and the base of the tail was shaved and the formulations were applied in semi-solid form for 9 days.

[0211] The formulations F1, F2, F3 used in the experiment are described in Table 2 (Embodiment 1).

[0212] Fluorescein In Figure 1 (fluorescein), there is a strong positive response visible in the dermis and skin layers in the form of green luminescence, the strongest in group F3 (white arrow). In group F3, the border between lubricated and non-lubricated skin is visible (blue line). Hairs with characteristic blue luminescence (red arrow) are visible in the skin. There is no visible luminescence in the dermis in group F2. The positive response was observed only superficially (white arrow). Magnification 40x.

[0213] In Figure 2, a positive response can be seen in the form of green luminescence in the muscle adjacent to the site of fluorescent dye injection (arrow). No positive response in group F2. Magnification 100x.

[0214] In figure 3, we can see hepatocytes and a strong positive response (complete staining of hepatocytes and intercellular spaces) in group F3. Negative response in the other groups. Magnification 400x.

[0215] The use of this fluorescent dye best demonstrates the penetration of the substance through the skin into the muscle and liver. In the F1 group, a positive signal is observed in the epidermis and in the muscle attached to the skin. A positive response was also observed in the liver and from the base of the tail to the tailbone. An equally strong signal with a very clear border in the skin (arrow) as well as in the muscle and liver was observed in the F3 group.

[0216] In Figure 4 (acridine orange), there is a strong positive response in the form of green luminescence in the dermis and subcutaneous layers, the strongest in groups F1 and F3 (arrows). In group F3, a positive response occurred only on the surface of the epidermis. Magnification 40x.

[0217] In Figure 5 (muscle tissue), a positive response similar to that of the skin is visible and it was necessary to correct the image contrast to show the signal at low magnification. A negative response was observed in the F2 group (strong contrast enhancement of the image). Magnifications 100x, 100x and 400x.

[0218] In Figure 6 showing liver samples, a positive response is visible in group F3 (arrows). Negative results were observed in the other groups. Magnification 400x.

[0219] The use of this fluorescent dye as well as other rhodamines can be applied to the study of the penetration of substances into the body through the skin, provided that the passage and accumulation of the dye in the cells is described. The fluorescent dye is much better exposed in areas of high cell density (epithelium, muscle tissue, liver cells) and therefore less exposed in the skin or subcutaneous connective tissue.

[0220] In Figure 7, rhodamine very strongly stained glandular cells and other connective tissue cells in the dermis. The strongest positive response was observed in groups F1 and F3. Magnification 100x, 100x and 40x.

[0221] In Figure 8 showing liver samples, weak positive responses are visible in groups F1 and F3, magnification 100x.

[0222] Rhodamine showed a very high ability to stain skin elements such as epidermal gland cells (both sebaceous and sweat glands), dermal cells (fibroblasts, macrophages and lymphocytes) and, after crossing the blood barrier, was partially accumulated in the liver. Example 32

[0223] Process for the preparation of pharmaceuticals containing substances from the group of proteins Substances from the group of antibodies were used as reference substances:

[0224] Antibody to the CD3 antigen, a molecule present on T lymphocytes, is a protein complex that is part of the T lymphocyte receptor (TCR) and is made up of five chains: δ, γ, ε, ζ, and η. The CD3 chain mediates the signal transduction from the antigen-bound T lymphocyte receptor into the cell. CD3 is the equivalent of the Igα and Igβ molecules on B lymphocytes.

[0225] Each of the CD3 chains is encoded by a separate gene: 1. The CD3 gamma chain is encoded by the CD3G gene located on chromosome 11q23 in males; 2. The CD3 delta chain is encoded by the CD3D gene, which is also located on chromosome 11q23. 3. The CD3 zeta chain is encoded by the CD247 (formerly CD3Z) gene located on chromosome 1q22-q25.

[0226] As a result of alternative splicing, the CD3ζ chain can be produced in a form that contains fewer lymphocyte activation motifs and is called CD3η. This form of protein is produced mainly in thymocytes and partially replaces CD3ζ in the TCR complex. The mRNA for CD3η is also detected in mature T cells, but its content is much smaller (more than 10 times) than that of the mRNA for CD3η, and therefore, CD3η is thought to perform its function mainly during the development of T lymphocytes.

[0227] During the development of T lymphocytes in the thymus, CD3 protein expression can be found at the prothymocyte stage, but only in the cytoplasm of the cells. A similar situation occurs at the prothymocyte stage, but when expression of the TCRα and TCRβ chains begins, the CD3 molecule also appears in the cell membrane. Membrane (surface) expression of CD3 is characteristic of each further developmental stage of T lymphocytes and persists until the cell dies.

[0228] In addition to T cells, weak CD3 expression, located only in the cytoplasm, was observed in Purkinje cells.

[0229] The CD3γ, CD3δ and CD3ε peptide chains belong to the same immunoglobulin superfamily and show strong similarities in amino acid sequence and structure. They all have an extracellular immunoglobulin-like domain followed by a helical fragment that crosses the cell membrane and the cytoplasmic tail containing an activated ITAM sequence. Each of the γ, δ and ε chains contains two ITAM motifs. The CD3ζ chain, in turn, does not have an extracellular immunoglobulin domain but instead contains four ITAM motifs in the cytoplasmic part. The CD3η variant has three ITAM motifs. The transmembrane region of each CD3 molecule is negatively charged, leading to the formation of a complex with the positively charged TCR chain. Thus, the TCRα-CD3γ-CD3ε-CD3ζ-CD3ζ and TCRβ-CD3δ-CD3ε complexes are formed, which together form the complete TCR complex.

[0230] T lymphocytes are the cells that are crucially involved in the development of inflammation. For this reason, drugs whose action is presumed to inhibit CD3 (and therefore T lymphocyte) activation were tested for their anti-inflammatory properties in autoimmune diseases and type 1 diabetes.

[0231] Fluorescent dye or enzyme-conjugated monoclonal antibodies against CD3 allow the determination of the frequency or number of these cells in blood and tissues using cytometric and immunohistochemical methods.

[0232] In diagnosis, the determination of CD3 is particularly useful for leukemia.

[0233] The composition ratio of the F1 carrier described in Example 1 was used in the experiment: an amount of 100 mg of active substance (concentration of 50 μg / g of anti-CD3 antibody suspension) was added to an amount of 20 g of the carrier composition obtained according to the procedure described for F1 and melted at 40 degrees Celsius.

[0234] After mixing (200 RPM) for about 10 minutes, a homogenous mixture was obtained and the substances used were completely dissolved after mixing. The mixture thus obtained was added to the substrate (80 g) described in Example 4 with stirring (500 RPM) for 8 minutes.

[0235] As a result, a formulation of homogenous consistency and color was obtained with a uniformly powdered material and a ratio between carrier and substrate of 1:5. Example 33

[0236] Process for the preparation of pharmaceuticals containing substances from the group of proteins Substances from the group of antibodies were used as reference substances:

[0237] Antibodies against the CD3 antigen The composition ratio of the F1 carrier described in Example 1 was used in the experiment: an amount of 100 mg of active substance (concentration of 50 μg / g of anti-CD3 antibody suspension) was added to an amount of 1 g of the carrier composition obtained according to the procedure described for F1 and melted at 40 degrees Celsius.

[0238] After mixing (200 RPM) for about 10 minutes, a homogenous mixture was obtained and the substances used were completely dissolved after mixing. The mixture thus obtained was added to the substrate (98.8 g) described in Example 4 with stirring (500 RPM) for 8 minutes.

[0239] As a result, a formulation of homogenous consistency and color was obtained with a uniformly powdered material and a ratio between carrier and substrate of 1:99. Example 34

[0240] Process for the preparation of pharmaceuticals containing substances from the group of proteins Substances from the group of antibodies were used as reference substances:

[0241] Antibodies against the CD3 antigen The composition ratio of the F1 carrier described in Example 1 was used in the experiment. A portion of the substrate in an amount of 20 g described in Example 4 was added to a composition of the carrier in an amount of 20 g obtained according to the procedure described for F1 and melted at 40 degrees Celsius, followed by an amount of 100 mg of active substance (concentration of 25 μg / g of anti-CD3 antibody suspension).

[0242] After mixing (400 RPM) for about 10 minutes, a homogenous mixture was obtained and the substances used were completely dissolved after mixing. The mixture so obtained was added to the remaining substrate (20 g) described in Example 4 with stirring (500 RPM) for 8 minutes.

[0243] As a result, a formulation of homogenous consistency and color was obtained with a uniformly raised material, the ratio between carrier and substrate was 1:2, and the amount of carrier constituted 20% of the total weight of the final product. Example 35

[0244] Process for the preparation of pharmaceuticals containing substances from the group of proteins Substances from the group of antibodies were used as reference substances:

[0245] Antibodies against the CD3 antigen The composition ratio of the F1 carrier described in Example 1 was used in the experiment. A portion of the substrate in an amount of 1 g described in Example 4 was added to a composition of the carrier in an amount of 1 g obtained according to the procedure described for F1 and melted at 40 degrees Celsius, followed by an amount of 100 mg of active substance (concentration of 25 μg / g of anti-CD3 antibody suspension).

[0246] After mixing (400 RPM) for about 10 minutes, a homogenous mixture was obtained and the materials used were completely dissolved after mixing. The mixture so obtained was added to the remaining substrate (96 g) described in Example 4 with stirring (500 RPM) for 8 minutes.

[0247] As a result, a formulation of homogenous consistency and color was obtained with a uniformly raised material, the ratio between carrier and substrate was 1:97, and the amount of carrier constituted 1% of the total weight of the final product.

[0248] Experiments were carried out to confirm the functionality of the described solutions.

[0249] The goal of the study was to evaluate the skin penetration ability of anti-mouse CD3ε antigen labeled with Alexa Fluor 647 fluorescent dye encapsulated in a micellar carrier. Due to the specificity of the antibody encapsulated in the carrier, the study was performed on a mouse model.

[0250] 10 mg of the carrier with the antibody and the base (carrier without the antibody) were applied to the ears of the mice and mounted on slides using silicone ointment on the inside of each of the left and right ears. A uniform distribution of the carrier throughout the ear was achieved after mounting a cover slip. The described procedure was used for formulations A (F2+CD3), B (F1+CD3), C (amphiphilic substrate+CD3), D (lipophilic substrate+CD3) and control bases A0 (F2), B0 (f1), C0 (amphiphilic substrate) and D0 (lipophilic substrate). The formulations so prepared were incubated for 4 hours at 37°C. After this time, the cover slip was removed, the test carrier was removed, a new cover slip was mounted and images were recorded. For the first comparison of the formulations, imaging was performed with a Leica SP8 MP confocal microscope using a water lens with a magnification of 25x. Fluorescence emission in the range of 646-750 nm was collected for AF647 dye excited with a 638 nm laser (to visualize antibodies), 638 nm laser light reflection in the range of 624-653 nm (to visualize skin layers) and 440-459 nm for the second harmonic after excitation with an infrared laser with a wavelength of 900 nm (to visualize collagen in the dermis). Samples were scanned in the Z-axis to a depth of approximately 100 μm encompassing the entire thickness of the epidermis and dermis. Three randomly selected fields of view were imaged for each formulation.

[0251] FIG. 9 shows normalized values, where the mean fluorescence intensity of the test tissues is a percentage of the fluorescence intensity of control tissues treated with carrier without antibody (100%).

[0252] After an initial comparison of all the carriers and their controls, formulation B (F1+CD3) was selected for further analysis due to the highest difference (p=0.1) in the fluorescence intensity of the tissues treated with this carrier compared to the tissues treated with the base mixture. For the remaining carriers, the fluorescence intensity of the investigated tissues was not different compared to the control tissues, which may indicate that the carriers did not pass through the epidermal layer of the analyzed ears and did not penetrate deep into the dermis.

[0253] After initial evaluation, formulation B (F1+CD3) was tested against more tissues (n=3).

[0254] The data are presented in chart form: Figure 10 shows the absolute values ​​of the mean fluorescence intensity for carrier B (F1+CD3) and B0 (F1) basal mixture, while Figure 11 shows the normalized values, where the mean fluorescence intensity of tissues treated with carrier B (F1+CD3) is a % of the fluorescence intensity of tissues treated with the basal mixture (100%).

[0255] For the initial comparison of the mean values ​​determined for the four series of results, A, B, C and D, the non-parametric Kruskal-Wallis test of variance (ANOVA) was used together with the median test and Dunn's test for non-parametric post-hoc peer-to-peer comparisons (Table 4 and Figure 12).

[0256] [Table 26]

[0257] Based on the results obtained, the data obtained for series B were subjected to further statistical analysis. The mean values ​​of the variables Prep_B_base and Prep_B_AF647 calculated from the 13 repetitions were compared by parametric Student's t-test. The normality of distribution for the analyzed variables was assessed by the Kolmogorov-Smirnov, Lilliefors and W Shapiro-Wilk tests. The Levene and Brown-Forsyth tests were used to assess their homogeneity of variance. For all statistical analyses and tests used, the significance level was α=0.05 or its value corrected by Bonferroni correction in case of multiple comparisons.

[0258] The results of the study are illustrated in FIGS.

[0259] [Table 27]

[0260] Conclusion: After use of formulation B, which contains an antibody against the mouse CD3ε antigen labeled with Alexa Fluor 647 fluorescent dye, the fluorescence intensity increases by approximately 40% compared to the control without antibody.

[0261] For the remaining formulations, no differences were observed between vehicle with antibody and vehicle control treated tissues.

Claims

1. A pharmaceutical carrier for a dosage form for transporting an active agent through the skin (excluding those containing water) comprising a solvating aprotic solvent, a carbonic acid amide, and an unsaturated fatty acid-containing vegetable oil, The aprotic solvent is present in an amount of 0.01% to 5% by weight of the carrier; The carbonic acid amide is present in an amount of 0.01% to 2% by weight of the carrier; The unsaturated fatty acid-containing vegetable oil is contained in an amount of 1% to 5% by weight of the carrier, the aprotic solvent is dimethylsulfoxide; the carbonic acid amide is urea, The vegetable oil is selected from the group consisting of corn oil, linseed oil, rapeseed oil, peanut oil, sunflower oil, hempseed oil and other vegetable oils containing unsaturated fatty acids; An anhydrous carrier characterized by:

2. 10. An anhydrous pharmaceutical composition comprising the carrier of claim 1 and an active agent, wherein the pharmaceutical composition is in a form for delivery through the skin, the weight ratio of the active agent to the carrier is from 1:10 to 2:1, and the carrier comprises from 1% to 20% by weight of the composition.

3. 3. The anhydrous pharmaceutical composition according to claim 2, containing excipients used for preparing said anhydrous pharmaceutical composition, characterized in that the weight ratio of said active substance to said excipients is 1:2.5 to 1:

494.

4. 4. An anhydrous pharmaceutical composition according to claim 2 or 3, characterized in that the active substance is selected from active substances of natural or synthetic origin.

5. 5. The anhydrous pharmaceutical composition according to claim 3 or 4, characterized in that the excipient components are selected from lard, duck fat, goose fat, Eucerin™, Lekobaza™, Hascobaza™, white petrolatum and yellow petrolatum, and any combination thereof.

6. An anhydrous pharmaceutical composition described in any one of claims 2 to 5, characterized in that the dosage form is an ointment or a semi-solid mixture.

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