EA043805B1 - NEW APPLICATION OF SULPHIC ACID SALTS OF CHOLECALCIFEROL AND NEW SULFULIC ACID SALTS OF CHOLECALCIFEROL - Google Patents

Description

The present invention relates to the use of pharmaceutically acceptable sulphates of cholecalciferol to combat vitamin D ₃ deficiency in vertebrates and especially in humans by introducing into the body a method that ensures the entry of sulphates of cholecalciferol in an amount of at least 5% as such without metabolization from the place of their introduction into the systemic fluid transport system in the human body, to new pharmaceutically acceptable sulfate salts of cholecalciferol, as well as to pharmaceutical compositions for the above administration containing pharmaceutically acceptable sulfate salts of cholecalciferol.

It is known that a significant part of the world's population suffers from a lack of vitamin D ₃ . In the human body, vitamin D ₃ is known to be formed naturally in the skin under the influence of ultraviolet light in the form of its water-soluble sulfate (cholecalciferol sulfate) from 7-dehydrocholesterol. However, for many people, especially during the winter months, it is not possible to receive enough UV light to allow vitamin D3 sulfate to form in the skin in sufficient quantities.

Cholecalciferol sulfate is found in human milk and in small quantities also in other milk. Some other foods, such as fish oil, contain non-sulfated fat-soluble vitamin D ₃ (cholecalciferol). In dietary supplements, vitamin D ₃ is also present in the form of fat-soluble cholecalciferol. This last-mentioned form of vitamin D ₃ is not sulfated in the body.

The present invention was based on the objective of providing natural cholecalciferol sulfate into the body in a manner that provides rapid onset of action or longer-lasting action, without the need to irradiate the skin with ultraviolet light.

Summary of the invention

Accordingly, the invention proposes the use of pharmaceutically acceptable cholecalciferol sulphate salts for combating vitamin D3 deficiency in vertebrates and especially in humans by introducing into the body a method that ensures that cholecalciferol sulphate salts are delivered in an amount of at least 5% as such without metabolization from the place of their introduction into the systemic fluid transport system in the human body, new pharmaceutically acceptable sulfate salts of cholecalciferol, as well as pharmaceutical compositions for the above administration containing pharmaceutically acceptable sulfate salts of cholecalciferol.

The invention further provides new pharmaceutically acceptable sulfate salts of cholecalciferol, especially sulfate salt of cholecalciferol and magnesium, sulfate salt of cholecalciferol and calcium and sulfate salt of cholecalciferol and L-lysine.

The invention further provides a pharmaceutical composition containing at least one pharmaceutically acceptable sulfate salt of cholecalciferol and a carrier suitable for transdermal or transmucosal administration or for intradermal, subcutaneous or intramuscular injection.

Brief description of graphic materials

The only drawing attached to the description shows the NMR spectrum (200 MHz, internal standard: TMS) of cholecalciferol sulfate and L-lysine in D ₄ -methanol.

Detailed Description of the Invention

When creating the invention, it was unexpectedly found that the intake of sulfate salts of cholecalciferol, and thereby vitamin D3 in its natural (native) form, into the body can be ensured in a certain way without their immediate metabolization to water-insoluble cholecalciferol or (25OH)-cholecalciferol. The lymphatic system is capable of distributing water-soluble sulfate salts of cholecalciferol as such in the body. Water-soluble sulfate salts of cholecalciferol are, of course, soluble in the blood, but under certain circumstances they can undergo rapid, for example enzymatic, decomposition in it.

By combating vitamin D ₃ deficiency according to the present invention is meant the prevention and treatment of vitamin D ₃ deficiency.

Pharmaceutically acceptable sulphate salts or sulfates of cholecalciferol according to the present invention are understood to mean salts in which their cation is not toxic to the human body under any conditions. Pharmaceutically acceptable cations include, for example, Na, Mg, Ca, Zn, ammonium and protonated amino acids such as protonated lysine.

Sulfate salts of cholecalciferol are usually soluble in water or in a water-alcohol mixture. Examples of pharmaceutically acceptable cholecalciferol sulfate salts with an inorganic cation suitable according to the invention include cholecalciferol sodium sulfate, cholecalciferol magnesium sulfate, cholecalciferol calcium sulfate and cholecalciferol ammonium sulfate. Examples of suitable pharmaceutically acceptable sulfate salts of cholecalciferol with an organic cation include sulfate

- 1 043805 salt of cholecalciferol and trimethylammonium and sulfate salt of cholecalciferol and L-lysine.

The method of administration proposed in the invention is chosen in such a way that it ensures that the administered cholecalciferol sulfate salt reaches at least 5% as such without metabolization from the site of its administration into the systemic fluid transport system in the human body. In a more preferred embodiment, the administered sulfate salt of cholecalciferol enters as such without metabolization from the site of its administration into the systemic fluid transport system in the human body in an amount that, depending on the method of administration, is at least 10, 15, 20, 25, 30, 35. 40, 45 or 50%.

In a preferred embodiment, the administered cholecalciferol sulfate salt is chemically altered (metabolized) in at least one systemic fluid transport system selected from blood and lymph within at least 5 minutes, more preferably within at least 10, 15, 20 or even 30 minutes, by less than 70%, preferably by less than 60%, for example by 50, 40, 30, 20 or even by 10% or less.

The systemic transport systems for fluids in the human body are, for example, blood (circulatory system) and preferably lymph (lymphatic system).

The preferred methods of administration of pharmaceutically acceptable sulfate salts of cholecalciferol are transdermal and transmucosal administration, as well as intradermal, subcutaneous and intramuscular injections, including injections of prolonged-release preparations with slow release and absorption of the active substance (depot injections). Oral administration (ie, ingestion into the stomach, for example by swallowing), as well as intravenous and intra-arterial injections are expressly excluded as routes of administration.

The molar amount of administered sulfate salts of cholecalciferol generally corresponds to the molar amount recommended for cholecalciferol.

Cholecalciferol sulphate salts can be prepared by converting commercially available cholecalciferol by reacting it with a sulfur trioxide-pyridine complex in pyridine and then converting it with triethylamine into cholecalciferol triethylammonium sulphate, which can then be reprecipitated in aqueous solution by adding saturated solutions of suitable cations to give salts with these cations.

Alternatively, cholecalciferol can be converted by reaction with the sulfur trioxide-pyridine complex in pyridine and then directly converted by reaction with the desired cation, optionally in the presence of a suitable buffer, into the desired salt.

The pharmaceutical compositions of the invention contain a suitable carrier. In addition, they may also contain additional active ingredients, such as other vitamins, minerals and trace elements, as well as any type of medicine.

Suitable carriers for transdermal administration include all carriers known to those skilled in the art of pharmacy that are used for this purpose. These include liquid or solid carriers with a fat or oil base, as well as aqueous or hydroalcohol-based carriers. Dosage forms can be presented in the form of ointments and oils, lotions, solutions that can be sprayed or sprayed, suspensions and emulsions of any type and patches containing the active substance in a carrier. Dosage forms may contain penetration enhancers, such as dimethyl sulfoxide, emulsifiers, as well as all sorts of other auxiliary substances common in pharmacy, such as diluents, flavoring agents, solubilizers, glidants, suspending agents, binders, preservatives.

Dosage forms suitable for transmucosal administration are, for example, aqueous or hydroalcoholic solutions, which may contain additional excipients, fat-based suppositories, creams, gels, pastes, foams or aerosols suitable for vaginal use and pessaries and tampons, as well as solid dosage forms suitable for topical oral or sublingual use, including lozenges which contain the active ingredient in a flavored base, usually sucrose and gum arabic or tragacanth, lozenges which contain the active ingredient in an inert base such as gelatin or glycerin, and chewing gum. Dosage forms for transmucosal administration may also contain penetration enhancers, such as dimethyl sulfoxide, emulsifiers, as well as all sorts of other excipients common in pharmacy.

Preparations (forms of preparation) in liquid form for parenteral administration by intradermal, subcutaneous and intramuscular injections include suspensions, solutions or emulsions in oily or aqueous carriers (bases). Such preparations may contain auxiliary substances such as suspending agents, stabilizers and/or dispersants. Alternatively, the active substance may be presented in powder form, which is mixed with a suitable carrier, such as sterile pyrogen-free water, before use to prepare the final formulation.

A detailed review of forms of application that are suitable for use in the present invention is given, for example, in Remington: The Science and Practice of Pharmacy, ed. Allen, Loyd V. Jr., 22nd ed., Pharmaceutical Press.

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Examples

Example 1. Preparation of sulfate salt of cholecalciferol and triethylammonium.

1.21 g of a complex compound of sulfur trioxide and pyridine (approximately 6.76 mmol of pyridine in the form of a complex compound with SO ₃ , Sigma-Aldrich, SO ₃ content of at least 45 wt.% according to the manufacturer) and 1.21 g (3 .14 mmol) cholecalciferol (Sigma-Aldrich) was mixed under a nitrogen atmosphere with 6.5 ml of pyridine. The resulting solution was vigorously stirred for 1 hour at 58°C. 0.63 ml (0.456 g, 4.55 mmol) of triethylamine was then added and stirred for a further 20 min at 58°C.

The reaction mixture was then cooled using an ice bath at 0°C, 16.5 mL of cold methanol/trichloromethane (10:1 by volume) was added and stirred for 20 minutes.

The solution was filtered through a glass frit and the solvent was removed on a rotary evaporator. The residue for subsequent purification was mixed twice more with a mixture of methanol and trichloromethane (in a volume ratio of 10:1) and the solvent was removed on a rotary evaporator, resulting in 1.52 g (3.13 mmol, 99.7%) of cholecalciferol sulfate salt and triethylammonium.

TLC (silica gel): Rf=0.42 in a mixture of methanol and trichloromethane (volume ratio 1:9).

Example 2. Preparation of sulfate salt of cholecalciferol and ammonium.

1.52 g (3.13 mmol) cholecalciferol triethylammonium sulfate was mixed with approximately 14 ml of saturated ammonium acetate solution until the cholecalciferol ammonium sulfate precipitated into a white precipitate, which, after standing overnight, was then filtered by filtration through a glass frit and dried in high vacuum while cooling with tap water at a temperature of approximately 15°C.

Yield: 1.74 g (3.10 mmol, 99%).

Melting point: 104-108°C.

Example 3. Preparation of sulfate salt of cholecalciferol and sodium.

1.52 g (3.13 mmol) cholecalciferol triethylammonium sulfate was mixed with saturated sodium chloride solution (approximately 14 ml) until the cholecalciferol sodium sulfate precipitated into a white precipitate, which after standing overnight was then filtered by filtration through a glass frit and dried in a high vacuum while cooling with tap water at a temperature of approximately 15°C.

Yield: 1.75 g (3.10 mmol, 99%).

The NMR spectrum of the product was identical to that published by L.E. Reeve et al., The Journal of Biological Chemistry, 1981, vol. 256, No. 2, p.. 824.

Example 4. Preparation of sulfate salt of cholecalciferol and magnesium.

1.52 g (3.13 mmol) cholecalciferol triethylammonium sulfate was mixed with a saturated magnesium chloride solution (approximately 13 ml) until the cholecalciferol magnesium sulfate precipitated into a white precipitate, which after standing overnight was then filtered by filtration through a glass frit and dried in a high vacuum while cooling with tap water at a temperature of approximately 15°C.

Yield: 1.76 g (3.10 mmol, 99%).

Melting point: 107-110°C (with decomposition).

Example 5. Preparation of sulfate salt of cholecalciferol and calcium.

1.52 g (3.13 mmol) of cholecalciferol and triethylammonium sulfate were mixed with a saturated solution of CaCl ₂ -2H ₂ O (approximately 13 ml) until the cholecalciferol and calcium sulfate precipitated into a white precipitate, which after standing overnight was then filtered off by filtration through a glass frit and dried under high vacuum while cooling with tap water at a temperature of approximately 15°C.

Yield: 1.81 g (3.10 mmol, 99%).

TLC (silica gel): Rf=0.48 in a mixture of methanol and trichloromethane (1:9 volume ratio).

Melting point: 97-101°C (with decomposition).

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Example 6. Preparation of sulfate salt of cholecalciferol and L-lysine.

517.4 mg of a complex compound of sulfur trioxide and pyridine (Sigma-Aldrich, SO ₃ content of at least 45 wt.% according to the manufacturer) (approximately 2.90 mmol of pyridine in the form of a complex compound with SO ₃ ) and 506.3 mg ( 1.32 mmol) cholecalciferol (Sigma-Aldrich) was dissolved under nitrogen in 4.4 ml of pyridine and heated for 1 hour with vigorous stirring at 55°C. After adding 10 mL of an ice-cold mixture of methanol and trichloromethane (1:9 v/v), the solvents were removed using a rotary evaporator. Then 0.29 g of L-lysine (1.98 mmol) was added to 0.5 ml of sodium phosphate buffer solution pH 7.3 and stirred for 5 minutes. Then, 20 ml of an ice-cooled mixture of methanol and trichloromethane (in a volume ratio of 1:9) was added with stirring, after which the mixture was evaporated on a rotary evaporator. 10 ml of absolute ethanol was added to the residue and the solution was placed in a refrigerator overnight. Thereafter, the ethanol was decanted from the resulting white creamy precipitate and the residue was dried under cooling with tap water at about 15°C to obtain the title compound as a white powder (815 mg, 1.30 mmol, 98.5%).

TLC (silica gel): Rf=0.33 in a mixture of methanol and trichloromethane (volume ratio 1:9).

Melting point: 168°C (with decomposition).

NMR spectrum: see drawing attached to the description.

Example 7. Preparation of sulfate salt of cholecalciferol and D,L-lysine.

Cholecalciferol sulfate and D,L-lysine were prepared similarly to the preparation of cholecalciferol sulfate and L-lysine, using D,L-lysine (Sigma-Aldrich) instead of L-lysine.

The NMR spectrum in D ₄ -methanol confirmed the structure of the resulting product.

Example 8. Transdermal dosage form of the sulfate salt of cholecalciferol and L-lysine in an oil base.

100 mg of sulfate salt of cholecalciferol and L-lysine was mixed with 11.04 ml of oleic acid, after which 0.83 ml of dimethyl sulfoxide was added. The mixture was then stirred with a magnetic stirrer at room temperature (21-25°C) for 2 days. Next, 10 ml of glycerol trioleate and 7 ml of glycerol monooleate (Pecerol®, Gattefosse) were added and the mixture was shaken vigorously. After waiting until the mixture became foamless, an almost completely transparent stable solution of the sulfate salt of cholecalciferol and L-lysine in an oil base was obtained.

Example 9 Application of the transdermal dosage form of Example 8 to the skin.

Approximately 2 ml of the dosage form prepared according to example 8 was applied in a thin layer to the skin of the test subject and left to absorb for 4 hours. Then the skin was thoroughly wiped with a sterile cotton swab soaked in 96% ethanol. The cotton swab was boiled with additional 96% ethanol and wrung out, after which the ethanol was evaporated on a rotary evaporator and a small amount of a mixture of chloroform and methanol (in a volume ratio of 9:1) was added to the flask. TLC analysis of this mixture on silica gel showed that practically no amount of cholecalciferol sulfate and L-lysine remained on the skin (R _f : 0.33).

Example 10. Preparation of aqueous solutions for injection.

To prepare a 2 ml injection solution containing cholecalciferol sulfate in an amount that corresponds to 10,000 IU of vitamin D ₃ :

a) 1.5695 mg of sulfate salt of cholecalciferol and sodium for the preparation of solution A,

b) 1.6243 mg of sulfate salt of cholecalciferol and calcium for the preparation of solution B and

c) 1.970 mg of sulfate salt of cholecalciferol and L-lysine for the preparation of solution B was dissolved in each case in 10 ml of distilled water.

Then, 89.9028 mg of sodium chloride was added to each solution to make it isotonic.

The pH value of solution B was adjusted to 7 using 0.5N. NaOH.

The solutions were then sterilized under argon by filtration through a membrane with a pore size of 0.22 μm and filled into 2-ml ampoules.

The information disclosed in all publications cited herein, such as patent specifications, published patent applications, journal articles and books, is incorporated herein by reference in its entirety.

Claims (8)

1. Pharmaceutically acceptable sulfate salt of cholecalciferol and L-lysine. 2. A method for treating or preventing vitamin D ₃ deficiency, including introducing into the body of vertebrate animals and humans a pharmaceutically acceptable sulfate salt of cholecalciferol at π. 1 and a carrier suitable for transdermal or transmucosal administration or for intradermal, subcutaneous or intramuscular injection. 3. The method according to claim 2, where the injections are injections of long-acting drugs with a slow release of the active substance. 4. The method of claim 2, wherein the carrier is an oil- or fat-based carrier suitable for transdermal administration. 5. The method according to claim 2, wherein the carrier is an aqueous or aqueous-alcohol based carrier suitable for transdermal administration. 6. Method according to one of claims 2-5, where the pharmaceutically acceptable sulfate salt of cholecalciferol and the carrier are presented in the form of a patch. 7. The method according to claim 2, wherein the carrier is an aqueous or aqueous-alcohol based carrier suitable for transmucosal administration. 8. The method according to claim 2, where the carrier is an aqueous or aqueous-alcohol based carrier suitable for intradermal, subcutaneous or intramuscular injection.