Cholecalciferol sulfate and its use for the treatment of vitamin D deficiency

Abstract

To achieve supplying of a body with natural cholecalciferol sulfate in a rapid or otherwise prolonged fashion without the necessity for irradiating the skin with UV radiation.SOLUTION: Pharmaceutically acceptable cholecalciferol sulfate salts are used for combating vitamin D3 deficiency. The invention also relates to cholecalciferol sulfate salts and to pharmaceutical compositions of the pharmaceutically acceptable cholecalciferol sulfate salts.SELECTED DRAWING: None

Description

[Technical field]

[0001] The present invention relates to a method for the administration of vitamin D3 in vertebrates, particularly humans, by a method in which at least 5% of cholecalciferol sulfate is delivered to the systemic fluid transport system of the human body from the administration site without metabolism. 3 The present invention relates to the use of pharma- ceutically acceptable cholecalciferol sulfate to combat deficiency, to new pharma-ceutically acceptable cholecalciferol sulfate, and to pharmaceutical compositions for the above-described administration in dosage forms containing pharma-ceutically acceptable cholecalciferol sulfate. [Background technology]

[0002] A large proportion of the world's population does not have vitamin D 3 It is known that humans suffer from vitamin D deficiency under natural conditions. 3 Exposure of the skin to UV radiation converts 7-dehydrocholesterol into water-soluble vitamin D 3 Vitamin D is known to be produced in humans as a sulfate ester (cholecalciferol sulfate). Many people do not obtain sufficient amounts of vitamin D in their skin, especially during the winter months. 3 They cannot expose themselves to enough UV radiation to form sulfate esters.

[0003] Cholecalciferol sulfate is found in breast milk and in small amounts in other types of milk. Some other types of foods, such as cod liver oil, contain non-sulfated fat-soluble vitamin D. 3 (Cholecalciferol). In dietary supplements, Vitamin D 3 Vitamin D is also found in the form of fat-soluble cholecalciferol. 3 This latter form is not sulfated in the body. [Prior art documents] [Non-patent literature]

[0004] [Non-Patent Document 1] Remington, The Science and Practice of Pharmacy, edited by Allen, Loyd V. Jr., 22nd edition, Pharmaceutical Press. [Non-Patent Document 2] LE Reeve et al., The Journal of Biological Chemistry (1981), Vol. 256, No. 2, p. 824 Summary of the Invention [Problem to be solved by the invention]

[0005] The object of the present invention was to provide the body with natural cholecalciferol sulfate in a rapid or otherwise long-term manner, without the need to expose the skin to UV radiation. [Means for solving the problem]

[0006] Thus, the present invention provides a method for the administration of vitamin D3 in vertebrates, particularly humans, by an administration method in which at least 5% of cholecalciferol sulfate reaches the systemic fluid transport system of the human body from the administration site by itself without metabolism. 3 The present invention relates to the use of pharma- ceutically acceptable cholecalciferol sulfate to combat deficiency.

[0007] Furthermore, the present invention relates to novel pharma- ceutical acceptable cholecalciferol sulfates, in particular magnesium cholecalciferol sulfate, calcium cholecalciferol sulfate, and L-lysine-cholecalciferol sulfate.

[0008] Finally, the present invention relates to a pharmaceutical composition comprising at least one pharma- ceutically acceptable cholecalciferol sulfate and a carrier suitable for transdermal or transmucosal administration, or for intradermal, subcutaneous or intramuscular injection. [Brief description of the drawings]

[0009] [Figure 1] FIG. 1 shows the NMR spectrum (200 MHz; internal standard: TMS) of cholecalciferol sulfate L-lysine in D4-methanol. DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS

[0010] Surprisingly, cholecalciferol sulfate, and therefore vitamin D in its natural form, 3 It has been found that water-insoluble cholecalciferol or (25OH)-cholecalciferol can be supplied to the body in such a way that they are not immediately metabolized. The lymphatic system can distribute the water-soluble cholecalciferol sulfate in the body by itself. The water-soluble cholecalciferol sulfate is of course soluble in the blood. However, they are potentially subject to rapid, e.g. enzymatic, degradation.

[0011] As used herein, vitamin D 3 Combating Deficiency with Vitamin D 3 Prevention and treatment of deficiency.

[0012] Pharmaceutically acceptable cholecalciferol sulfate salts herein mean that the cations of these salts are in no way toxic to the human body. Pharmaceutically acceptable cations are, for example, Na, Mg, Ca, Zn, ammonium, and protonated amino acids such as protonated lysine.

[0013] Cholecalciferol sulfate is usually soluble in water or aqueous alcohol mixture. Examples of pharma- ceutically acceptable cholecalciferol sulfate with inorganic cations are cholecalciferol sodium sulfate, cholecalciferol magnesium sulfate, cholecalciferol calcium sulfate, and cholecalciferol ammonium sulfate. Examples of pharma-ceutically acceptable cholecalciferol sulfate with organic cations are cholecalciferol trimethylammonium sulfate and cholecalciferol L-lysine sulfate.

[0014] The administration method according to the present invention is selected such that at least 5% of the administered amount of cholecalciferol sulfate is transported from the administration site to the systemic fluid transport system of the human body without metabolism. Depending on the administration method, preferably at least 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, or 50% of the administered cholecalciferol sulfate reaches the systemic fluid transport system of the human body from the administration site by itself without metabolism.

[0015] Preferably, the administered cholecalciferol sulfate is chemically modified (metabolized) less than 70%, preferably less than 60%, for example less than 50%, 40%, 30%, 20%, or even less than 10%, or less in at least one systemic fluid transport system selected from blood and lymph over a period of at least 5 minutes, preferably at least 10, 15, 20, or even more than 30 minutes.

[0016] The systemic fluid transport systems of the human body are, for example, blood and preferably lymph.

[0017] The preferred modes of administration of pharma- ceutically acceptable cholecalciferol sulfate are transdermal or transmucosal administration, as well as intradermal, subcutaneous and intramuscular injections, depot injections, including sustained release of the active substance. Oral administration (i.e., delivered to the stomach by swallowing), intravenous and arterial injections are expressly excluded as modes of administration.

[0018] The molar amount of cholecalciferol sulfate administered generally corresponds to the molar amount recommended for cholecalciferol.

[0019] Cholecalciferol sulfate can be prepared by reacting commercially available cholecalciferol with pyridine-sulfur trioxide complex in pyridine, followed by subsequent reaction with triethylamine to give triethylammonium cholecalciferol sulfate, which can then be precipitated in aqueous solution by adding saturated solutions of the appropriate cations to form salts with these cations.

[0020] Alternatively, cholecalciferol can be reacted with pyridine-sulfur trioxide complex in pyridine and then reacted directly with the desired cation, optionally in the presence of a suitable buffer, to form the desired salt.

[0021] The pharmaceutical compositions according to the invention comprise a suitable carrier or a suitable vehicle. These may also contain additional active agents, such as other vitamins, minerals, and trace elements, as well as any type of medicinal agent.

[0022] Vehicles or carriers suitable for transdermal administration are all vehicles known to those skilled in the art of pharmacy for this purpose.These include liquid or solid vehicles based on fat or oil, and vehicles based on water or water-alcohol.The formulations can take the form of ointments and oils, lotions, solutions adapted to be sprayed, any kind of suspension and emulsion, and patches containing the active substance in the vehicle.The formulations can include penetration enhancers such as dimethylsulfoxide, emulsifiers, and any further additives commonly used in pharmacy, such as excipients, flavorings, solubilizers, lubricants, suspending agents, binders, and preservatives.

[0023] Formulations suitable for transmucosal administration are, for example, aqueous or alcoholic solutions, fatty-based suppositories, creams, gels, pastes, foams or sprays, which may contain further additives, as well as pessaries and tampons suitable for vaginal administration, and solid dosage forms suitable for local administration in the mouth or sublingual administration, including lozenges containing the active ingredient in a flavored base, usually sucrose and acacia gum or tragacanth; pastilles containing the active ingredient in an inert base such as gelatin or glycerol; and chewing gum. Formulations for transmucosal administration may also contain penetration enhancers, such as dimethylsulfoxide, emulsifiers, and any further additives common in pharmacy.

[0024] Liquid form formulations for parenteral administration by intradermal, subcutaneous and intramuscular injection include suspensions, solutions, or emulsions in oily or aqueous vehicles. The formulations may contain formulatory additives, such as suspending, stabilizing and / or dispersing agents. Alternatively, the active ingredient may be in powder form for mixing with a suitable vehicle, such as sterile pyrogen-free water, for constitution before use.

[0025] A detailed listing of suitable dosage forms for use in the present invention can be found, for example, in Remington, The Science and Practice of Pharmacy, edited by Allen, Loyd V. Jr., 22nd Edition, Pharmaceutical Press. EXAMPLES

[0026] Example 1 Preparation of cholecalciferol triethylammonium sulfate 6.5 ml of pyridine was dissolved in 1.21 g of pyridine-sulfur trioxide complex (approximately 6.76 mmol, SO 3 Pyridine complexed with Sigma-Aldrich; manufacturer's information: ≥ 45 wt.-% SO 3 ) and 1.21 g (3.14 mmol) cholecalciferol (Sigma-Aldrich). The resulting solution was stirred vigorously for 1 h at 58° C. Then 0.63 ml (0.456 g; 4.55 mmol) triethylamine was added and stirring was continued for another 20 min at 58° C.

[0027] The reaction mixture was then cooled in an ice bath at 0° C., 16.5 ml of cold methanol-trichloromethane (10:1 vol. / vol.) solution was added, and stirring was continued for 20 min.

[0028] The solution was filtered through a glass frit and the solvent was removed by rotary evaporation. For further purification, the residue was treated twice with a methanol-trichloromethane (10:1 Vol. / Vol.) solution and the solvent was removed by rotary evaporation, thereby obtaining 1.52 g (3.13 mmol; 99.7%) of cholecalciferol triethylammonium sulfate.

[0029] TLC (silica gel): R F = 0.42 in methanol-trichloromethane (1:9 vol. / vol.).

[0030] Example 2 Preparation of cholecalciferol ammonium sulfate About 14 ml of saturated ammonium acetate solution was added to 1.52 g (3.13 mmol) of triethylammonium cholecalciferol sulfate until a white precipitate of ammonium cholecalciferol sulfate was formed, which was left to stand overnight, then filtered using a glass frit and dried under high vacuum with cooling with tap water at a temperature of about 15° C. Yield: 1.74 g (3.10 mmol, 99%). Melting point: 104~108℃

[0031] Example 3 Preparation of cholecalciferol sodium sulfate

[0032] [ka]

[0033] A saturated solution of sodium chloride (about 14 ml) was added to 1.52 g (3.13 mmol) of triethylammonium cholecalciferol sulfate until a white precipitate of sodium cholecalciferol sulfate was formed, which was left to stand overnight, then filtered using a glass frit and dried under high vacuum while cooling with tap water having a temperature of about 15° C. Yield: 1.75 g (3.10 mmol, 99%).

[0034] The NMR spectrum was identical to that published by LE Reeve et al., The Journal of Biological Chemistry (1981), Vol. 256, No. 2, p. 824.

[0035] Example 4 Preparation of cholecalciferol magnesium sulfate A saturated solution of magnesium chloride (about 13 ml) was added to 1.52 g (3.13 mmol) of triethylammonium cholecalciferol sulfate until a white precipitate of magnesium cholecalciferol sulfate was formed, which was left to stand overnight, then filtered using a glass frit and dried under high vacuum while cooling with tap water having a temperature of about 15° C. Yield: 1.76 g (3.10 mmol; 99%). Melting point: 107-110°C (decomposition).

[0036] Example 5 Preparation of cholecalciferol calcium sulfate CaCl 2 2H 2 A saturated solution of O (about 13 ml) was added to 1.52 g (3.13 mmol) of triethylammonium cholecalciferol sulfate until a white precipitate of calcium cholecalciferol sulfate was formed, which was left to stand overnight, then filtered using a glass frit and dried under high vacuum while cooling with tap water having a temperature of about 15° C. Yield: 1.81 g, (3.10 mmol, 99%). TLC (silica gel): R f = 0.48 in methanol-trichloromethane (1:9 Vol. / Vol.) Melting point: 97-101°C (decomposition).

[0037] Example 6 Preparation of cholecalciferol sulfate L-lysine

[0038] [ka]

[0039] 517.4 mg of pyridine-sulfur trioxide complex (Sigma-Aldrich; ≥ 45 wt.-% SO according to manufacturer's information) 3 ) (approximately 2.90 mmol of SO 3 506.3 mg (1.32 mmol) of pyridine complexed with 1:1 ethanol and cholecalciferol (Sigma-Aldrich) were dissolved in 4.4 ml of pyridine under nitrogen atmosphere and heated at 55° C. for 1 h with vigorous stirring. After adding 10 ml of ice-cold methanol-trichloromethane (1:9 Vol / Vol) mixture, the solvent was removed on a rotary evaporator. 0.29 g (1.98 mmol) of L-lysine in 0.5 ml of sodium phosphate buffer, pH 7.3, was then added and stirred for 5 min. 20 ml of ice-cold methanol-trichloromethane (1:9 vol. / vol.) mixture was then added with stirring, after which the mixture was stripped on a rotary evaporator. 10 ml of absolute ethanol was added to the residue and the solution was stored overnight in the refrigerator. The ethanol was then decanted from the white creamy precipitate that formed and the residue was dried with cooling with tap water at a temperature of about 15° C., thereby yielding the title compound as a white powder (815 mg, (1.30 mmol); 98.5%). TLC (silica gel): R f = 0.33 in methanol-trichloromethane (1:9 Vol. / Vol.). Melting point: 168℃ (decomposition)

[0040] NMR spectrum: see Figure 1

[0041] Example 7 Preparation of cholecalciferol sulfate D,L-lysine Cholecalciferol sulfate D,L-lysine was prepared in a manner similar to that for cholecalciferol sulfate L-lysine, using DL-lysine (Sigma-Aldrich) instead of L-lysine.

[0042] D 4The structure is confirmed by NMR spectroscopy in methanol.

[0043] Example 8 Transdermal formulation of cholecalciferol sulfate L-lysine in an oil base 100 mg of cholecalciferol sulfate L-lysine was mixed with 11.04 ml of oleic acid, then 0.83 ml of dimethyl sulfoxide was added and stirred with a magnetic stirrer at room temperature (21-25°C) for 2 days. Subsequently, 10 ml of glycerol trioleate and 7 ml of glycerol monooleate (Pecerol®, Gattefosse) were added and vigorously shaken. After waiting until the mixture had disappeared, an almost completely clear stable solution of cholecalciferol sulfate L-lysine in the oil phase was obtained.

[0044] Example 9 Application of the transdermal formulation of Example 8 to the skin Approximately 2 ml of the formulation prepared in Example 8 was applied in a thin layer to the skin of the subject and allowed to penetrate for 4 hours. Subsequently, the skin was thoroughly wiped using a sterile cotton pad soaked in 96% ethanol. The cotton pad was boiled with additional 96% ethanol and squeezed. The ethanol was evaporated on a rotary evaporator and a small amount of chloroform-methanol (9:1 Vol. / Vol.) mixture was added to the flask. TLC of this mixture on silica gel revealed cholecalciferol sulfate L-lysine (R f : 0.33) was shown to be substantially absent on the skin.

[0045] Example 10 Preparation of aqueous injection solutions 10,000 IU of Vitamin D 3 To prepare 2 ml of injection solution containing cholecalciferol sulfate in an amount corresponding to: a) 1.5695 mg of sodium cholecalciferol sulfate to produce injection solution A b) 1.6243 mg of calcium cholecalciferol sulfate to produce injection solution B and c) 1.970 mg of cholecalciferol sulfate L-lysine to produce injection solution C were each dissolved in 10 ml of distilled water. 89.9028 mg of sodium chloride was then added to each of the solutions to make them isotonic.

[0046] The pH of Injection Solution B was adjusted to pH 7 using 0.5 N NaOH.

[0047] The solution was then filter-sterilized through a 0.22 μm membrane under an argon atmosphere and filled into 2 ml vials.

[0048] The disclosures of all documents cited herein, including, for example, patents, published patent applications, articles published in journals and books, are hereby incorporated by reference in their entirety.

Claims

[Claim 1] A pharmaceutical composition comprising a pharma- ceutical acceptable salt of cholecalciferol sulfate, the method comprising administering the composition to a vertebrate animal such that at least 5% of the salt of cholecalciferol sulfate enters the systemic fluid transport system of the human body from the site of administration without being metabolized. 3 To combat deficiencies, the method of administration is selected from transdermal and transmucosal administration, and intradermal, subcutaneous and intramuscular injections, including sustained drug release depot injections; A pharmaceutical composition, wherein the salt of cholecalciferol sulfate is cholecalciferol sulfate L-lysine.

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