EA010437B1 - Pharmaceutical compositions c0mprising active vitamin d compounds - Google Patents

Abstract

Disclosed are pharmaceutical compositions comprising an active vitamin D compound in emulsion pre-concentrate formulations, as well as emulsions and sub-micron droplet emulsions produced therefrom. The compositions comprise a lipophilic phase component, one or more surfactants, and an active vitamin D compound. The compositions may optionally further comprise a hydrophilic phase component.

Description

The present invention relates to new pharmaceutical compositions containing a compound of active vitamin Ό, and these pharmaceutical compositions are pre-concentrates of emulsions. The invention also relates to emulsions and to submicron emulsions (with droplet size less than a micron) obtained by diluting the emulsion pre-concentrates with an aqueous solution.

State of the art

Vitamin Ό is a fat-soluble vitamin that is indispensable as a positive regulator of calcium homeostasis. , E. Vgaipeta1b e! A1., (Ebch.), MsSga ^ -NS, Youe Wogk (1987)). The active form of vitamin Ό is 1a, 25-dihydroxyvitamin Ό ₃ , also known as calcitriol. Specific nuclear receptors of active vitamin соединений compounds were found in cells of various organs that are not involved in calcium homeostasis. (Msheg e! A1., Sapseg. Vech. 52: 515-520 (1992)). Compounds of active vitamin Ό not only affect calcium homeostasis, but also participate in osteogenesis (bone formation), modulation of immune responses, modulation of the process of insulin production by pancreatic B-cells, functioning of muscle cells and in the differentiation and growth of epidermal and hematopoietic tissues.

In addition, in the published literature there are many indications of the usefulness of the use of active vitamin соединений compounds in the treatment of cancer. For example, it was shown that some compounds of vitamin Ό and their analogues have a powerful antileukemic effect, since they stimulate the differentiation of malignant cells (especially leukemia cells) into benign macrophages (monocytes) and are useful in the treatment of leukemia (§iba e! A1., US patent No. 4391802, Raipbde e! A1., US patent No. 4594340). Also reported on the antiproliferative and differentiating effect of calcitriol and other analogues of vitamin Ό ₃ in connection with the treatment of prostate cancer (B1jor e! A1., US patent No. 5795882). Compounds of active vitamin Ό were also used in the treatment of skin cancer (Syba e! A1., Sapseg Vecheagsy 45: 5426-5430 (1985)), colon cancer (O1chtap e! A1., Sapseg Vechegs 47: 21-25 (1987)) and lung cancer (§а! о е! а1., Toyoki 1. Exp. Meb. 738: 445-446 (1982)). Other reports of the important therapeutic effect of active vitamin соединений compounds are summarized in US Pat. No. 6,043,079, General! a1.

Although ingestion of active vitamin соединений compounds can lead to very positive therapeutic results, the treatment of cancer and other diseases with these compounds is limited due to the effect of these compounds on calcium metabolism. At the dose levels necessary for the effective use of antioxidants as antiproliferative agents, active vitamin Ό compounds can cause a strong and potentially dangerous increase in blood calcium levels due to their inherent effect on calcium metabolism. That is, the clinical use of calcitriol and other active vitamin соединений compounds as antiproliferative agents is hindered or severely limited by the risk of hypercalcemia.

It has been shown that the problem of systemic hypercalcemia can be solved by taking a sufficient dose of the active vitamin соединения compound in a pulsed dosage regimen, so as to achieve an antiproliferative effect, but at the same time avoid the development of acute hypercalcemia (US Patent No. 6521608). In accordance with the set out in patent 6521608, the intake of active vitamin раз compounds can occur no more than once every three days, for example once a week, at a dose of at least 0.12 μg / kg per day (8.4 μg for a person weighing 70 kg). Pharmaceutical formulations used in pulsed dosage regimen in accordance with Patent 6521608 include 5100 μg of the active vitamin соединения compound and can be administered by oral, intravenous, intramuscular, topical, transdermal, sublingual, intranasal, intratumoral or other methods.

The trade name for calcitriol, manufactured by Vosie Láboga! Og1ech, is EIGHT. VLETIMA is supplied in the form of capsules containing 0.25 and 0.5 μg of calcitriol, and in the form of an oral solution containing 1 μg / ml of calcitriol. All dosage forms contain bottled hydroxyanisole (BHA) and bottled hydroxytoluene (BHT) as antioxidants. Capsules also contain fractionated coconut oil triglyceride, and oral solutions contain fractionated palm oil triglyceride (RyuchSchap'ch Eechk BeGegeps. 54 ^1st Ebyup, pp. 2649-2651, Meb1ca1 Esopotchch Sotrapu, 1ps1, 2000).

Calcitriol is known to be sensitive to light and readily oxidizes. In addition, calcitriol and other compounds of active vitamin Ό are lipophilic, i.e., readily soluble in lipids (fats) and some organic solvents, but practically insoluble or poorly soluble in water. Due to the lipophilic nature of the active vitamin соединений compounds, the dispersion of such compounds in aqueous solutions, in particular in gastric fluids, is quite limited. Accordingly, the pharmacokinetic parameters of the preparations of active vitamin соединений compounds described earlier in this technical field are suboptimal for taking large doses in a pulsed dosage regimen. In addition, the currently available preparations of active vitamin соединений compounds exhibit a rather strong scatter in the absorption values in the small intestine. Moreover, for oral administration, the ratio

- 1 010437 between the dose and blood concentration observed for most preparations of active vitamin соединений compounds, is non-linear, that is, the amount of the compound absorbed by the blood stream does not correlate with the amount of the compound taken in the dosage form, especially at high doses.

Thus, in the art there is a need to develop improved pharmaceutical formulations comprising active vitamin соединения compounds, in particular for the administration of large doses of the drug in a pulsed dosage regimen, designed to provide an antiproliferative (e.g., anti-cancer) effect, but not causing hypercalcemia. In particular, in the art there is a need to develop an improved pharmaceutical composition containing an active vitamin соединение compound that is stable for a long time even at elevated temperatures, and at the same time exhibits improved pharmacokinetic parameters with respect to the active vitamin соединения compound and a reduced absorption dispersion when taken by the patient.

SUMMARY OF THE INVENTION

In the present invention, the disadvantages previously existing in the art are overcome by developing a pharmaceutical composition comprising active vitamin соединения compounds in the form of emulsion pre-concentrate preparations. The pharmaceutical compositions in accordance with the present invention have advantages over known analogues, since they provide the preparation of dosage forms of active vitamin соединений compounds, for example, calcitriol, which have sufficiently high concentrations for ease of use, are stable and well dispersible in solution, and at the same time meet the necessary criteria pharmacokinetic parameters, in particular when taken in a pulsed dosage regimen. More specifically, in a preferred embodiment, the pharmaceutical compositions of the present invention provide a Stax 1.5-2 times greater than the C _max observed for RED and a shorter T _max than the same value observed for RED.

The emulsion preconcentrates of the present invention are non-aqueous preparations of the active vitamin витами compound capable of forming a pharmaceutically acceptable emulsion upon contact with water or other aqueous solutions.

Thus, the present invention provides pharmaceutical compositions that contain: (a) a component of the lipophilic phase, (b) one or more surfactants, and (c) a compound of active vitamin Ό, wherein the composition can be used as an emulsion pre-concentrate, which, when diluted with water in a water / composition ratio of approximately 1: 1, or with a higher water content, forms an emulsion having an optical density of more than 0.3 at 400 nm. Accordingly, the pharmaceutical compositions may also contain a hydrophilic phase component.

According to one aspect of the present invention, there is provided a liquid or semi-solid pharmaceutical composition comprising an active vitamin витами compound, about 50% lipophilic component, about 50% surfactant, about 0.05 to about 0.35% butylated hydroxyanisole (BHA), and from about 0.05 to about 0.35% butylated hydroxytoluene (BHT). In a preferred embodiment of the present invention, said active vitamin соединение compound is calcitriol. In addition, said component of the lipophilic phase is MUOUR 812. Preferably, said surfactant is Vitamin E TPC8. In a preferred embodiment, the pharmaceutical composition comprises a compound of active vitamin Ό, about 50% MULOUM 812 and about 50% vitamin E TRC8.

In accordance with another aspect of the present invention, there is provided a unit dosage form comprising a liquid or semi-solid pharmaceutical composition comprising a compound of active vitamin Ό, about 50% MULO 812, about 50% vitamin E TRC8, about 0.05 to 0.35% BHA, and from about 0.05 to about 0.35% OSH. In a preferred embodiment, said unit dosage form contains calcitriol as the active vitamin compound. In another preferred embodiment, the unit dosage form contains from about 10 to about 75 μg of calcitriol, preferably 45 μg of calcitriol. In yet another preferred embodiment, the unit dosage form comprises a component of the lipophilic phase MULOUL 812. In yet another embodiment, the unit dosage form contains TPC8 vitamin E as a surfactant. In another embodiment, the unit dosage form contains about 50% MULOUM 812 and about 50% Vitamin E TPC8. In yet another embodiment, the unit dosage form contains about 45 μg of calcitriol, about 50% MULOUR 812, about 50% vitamin E TPC8, about 0.05 to about 0.35% BHA, and about 0.05 to about 0.35 % OSH.

In accordance with another aspect of the invention, the unit dosage form described above may be a capsule, preferably a gelatin capsule. In addition, in a preferred embodiment, the total volume of ingredients in the capsule is 10-1000 μl.

- 2 010437

Thus, the present invention allows to obtain a pharmaceutical emulsion composition containing water and an emulsion preconcentrate, wherein said preconcentrate contains: (a) a component of the lipophilic phase, (b) one or more surfactants, (c) a compound of active vitamin активного and, possibly also a component of the hydrophobic phase.

Emulsions obtained from pre-concentrates of emulsions in accordance with the present invention (when diluted with water) include both emulsions in the traditional sense of the term by those skilled in the art (i.e., organic phase dispersions in water) and submicron emulsions (i.e. dispersions of the organic phase in water, the average diameter of the dispersion particles in which is less than 1000 nm.)

Methods have also been developed for the preparation of emulsion pre-concentrates, including active vitamin соединения compounds. The methods covered by this aspect of the present invention include thoroughly mixing the active vitamin соединения compound, for example calcitriol, and the lipophilic phase component, as well as one or more surfactants, and possibly the hydrophilic phase component.

In accordance with another aspect of the present invention, methods are provided for treating and preventing hyperproliferative diseases, in particular cancer and psoriasis, said methods comprising administering an active vitamin соединения compound in the form of a liquid or semi-solid pharmaceutical composition or unit dosage form to a patient who needs it. In an alternative embodiment, the patient may take the active vitamin соединение compound in the form of an emulsion preconcentrate preparation or an emulsion preparation prepared by diluting with an appropriate amount of water an emulsion preconcentrate according to the present invention. In a preferred embodiment of this aspect of the present invention, the patient takes an active vitamin витами compound, for example, in a pulsed dosage regimen. In accordance with this aspect of the present invention, the patient takes a compound of active vitamin Ό in the form of a pharmaceutical composition, unit dosage form or preparation of an emulsion preconcentrate, for example, not more than once every three days at a dose of at least 0.12 μg / kg per day. In one embodiment, the administration of the pharmaceutical composition or unit dosage form according to the invention is carried out in a pulsed dosage regimen, wherein said pulsed dosage regimen includes a patient taking said composition no more than once every three days, preferably no more than once a week, more preferably not more than once every three weeks.

In addition, in one embodiment, the dosage regimen also includes the administration of one or more chemotherapeutic agents or radiotherapeutic agents / procedures. In a preferred embodiment, the administration of the active vitamin соединения compound is carried out at least 12 hours before the administration of said one or more chemotherapeutic agents or radiotherapeutic agents / procedures. In a most preferred embodiment, the administration of the active vitamin соединения compound is carried out preliminarily from 1 day to about 3 months before the administration of said one or more chemotherapeutic agents or radiotherapeutic agents / procedures. In another preferred embodiment, the administration of the active vitamin соединения compound is carried out simultaneously with the administration of the indicated one or more chemotherapeutic agents or radiotherapeutic agents / procedures. In yet another preferred embodiment, administration of the active vitamin соединения compound is continued after administration of said one or more chemotherapeutic agents or radiotherapeutic agents / procedures. In yet another embodiment, the administration of the active vitamin соединения compound is carried out after administration of the indicated one or more chemotherapeutic agents or radiotherapeutic agents / procedures. In another embodiment, the administration of the active vitamin соединения compound is carried out 1 day before the administration of said one or more chemotherapeutic agents or radiotherapeutic agents / procedures. In yet another embodiment, the active vitamin соединение compound and said one or more chemotherapeutic agents or radiotherapeutic agents / procedures are taken no more than once every three weeks.

Brief Description of Drawings

In FIG. Figure 1 shows a graphical dependence of the average concentration of calcitriol in the blood plasma of dogs on the time elapsed since administration for three different preparations of calcitriol at a dose of 1 μg / kg.

In FIG. 2A and 2B are graphical plots of the average concentration of calcitriol in a dog’s blood plasma versus time after administration of high doses of semi-solid preparation # 3 in males (FIG. 2A) and females (FIG. 2B).

In FIG. ZA and 3B present graphical dependences of the concentration of calcitriol in the blood plasma of dogs on time after taking the semi-solid preparation # 3, in males (Fig. 3A) and in females (Fig. 3B).

In FIG. 4A and 4B show graphical dependences of the average concentration of calcium in the blood serum of dogs after taking elevated doses of semi-solid preparation # 3 in males (Fig. 4A) and in females (Fig. 4B).

- 3 010437

In FIG. 5A-5C show graphical relationships between the concentration of calcitriol in plasma and the concentration of calcium in the blood serum of male dogs after taking semi-solid preparation # 3.

In FIG. Figure 6 shows a graphical representation of the average concentration of calcitriol in human blood plasma after taking a semi-solid preparation # 3 in a dose group.

DETAILED DESCRIPTION OF THE INVENTION

The present invention is directed to the development of pharmaceutical formulations comprising active vitamin соединения compounds in the form of emulsion pre-concentrate formulations. The compositions in accordance with the present invention can avoid or significantly reduce the occurrence of problems associated with the treatment of compounds of active vitamin Ό and which have hitherto been part of the art, such as poor pharmacokinetic parameters of the compound when taken by the patient.

It was found that the compositions in accordance with the present invention allow the preparation of semi-solid and liquid preparations containing compounds of active vitamin Ό in sufficiently high concentrations for convenient administration, for example, oral, and at the same time have improved pharmacokinetic parameters of the compound of active vitamin Ό. For example, in comparison with SCAN, compositions according to the present invention exhibit C _max 1.5-2 times higher than C _max observed for SCAN and shorter T _max than observed for SC. The pharmaceutical compositions in accordance with the present invention preferably provide 0 ^ equal to at least about 900 pg / ml of plasma, more preferably from about 900 to 3000 pg / ml of plasma, and even more preferably from about 1500 to 3000 pg / ml of plasma. In addition, the compositions in accordance with the present invention preferably provide a T _{max of} less than 6 hours, more preferably from about 1 to 3 hours, and even more preferably from about 1.5 to 2 hours. In addition, the compositions in accordance with the present invention preferably provide T1 / 2 less than 25 hours, more preferably from about 2 to 10 hours and even more preferably from about 5 to 9 hours

The term C _{max is} defined as the maximum concentration of the active vitamin соединения compound in blood serum after taking the medication. The term T _{max is} defined as the time during which the concentration reaches C _max . The term T _{1/2 is} defined as the time required to reduce the concentration of the active vitamin соединения compound in blood serum by half. The pharmacokinetic values indicated in this description are true for the entire population of recipients taking the active vitamin соединение compound, and not for individual recipients. Thus, the preferred pharmacokinetic parameters will not necessarily be implemented for a particular individual taking the composition in accordance with the present invention. However, when taking the composition in accordance with the present invention, a sufficiently large set of subjects, the pharmacokinetic parameters will approximately coincide with the values specified in this description.

In accordance with one aspect of the present invention, a pharmaceutical composition is prepared comprising: (a) a component of the lipophilic phase, (b) one or more surfactants, and (c) a compound of active vitamin Ό; wherein said composition is an emulsion preconcentrate which, when diluted with water at a water / composition ratio of approximately 1: 1, or with a higher water content, forms an emulsion having an optical density of more than 0.3 at 400 nm. In accordance with this aspect of the invention, the pharmaceutical composition may also contain a hydrophilic phase component.

In accordance with another aspect of the present invention, an emulsion pharmaceutical composition is prepared comprising water (or another aqueous solution) and an emulsion preconcentrate.

The term emulsion preconcentrate, as used herein, means a system capable of creating an emulsion in contact with, for example, water. The term emulsion used in this description means a colloidal dispersion, including water and organic components, including hydrophobic (lipophilic) organic components. The term emulsion includes both traditional emulsions, that is, systems commonly understood by those in the art as sub-micron emulsions, as defined below.

The term submicron droplet emulsion, as used herein, means a dispersion containing water and organic components including hydrophobic (lipophilic) organic components, with droplets or particles formed by organic components having an average maximum size of less than 1000 nm.

Submicron droplet emulsions are identified by one or more of the following characteristics. They are formed spontaneously or almost spontaneously if their components come into contact, that is, without a significant supply of energy, for example, in the absence of heating or without the use of intensive shear mixing equipment or other significant mixing.

Particles of submicron emulsion can be spherical, however, other structures, for example, liquid crystals with scaly, hexagonal or isotropic symmetry, are possible. In general, submicron emulsions include droplets or particles having a maximum size (per

- 4 010437 example, average diameter) from about 50 to 1000 nm, and preferably from about 200 to 300 nm.

The term pharmaceutical composition, as used herein, is to be understood as a composition in which the individual components or ingredients are pharmaceutically acceptable. For example, if the composition is intended for oral administration, then its components should be acceptable for oral administration, and if the composition is intended for topical administration, then its components should be acceptable for topical administration.

When diluted with water, the pharmaceutical compositions of the present invention typically form emulsions. The emulsion in accordance with the present invention is formed by dilution of the pre-emulsion concentrate with water at a water / composition ratio of approximately 1: 1 or with a higher water content. In accordance with the present invention, the ratio of water / composition may be, for example, from 1: 1 to 5000: 1. In particular, the water / composition ratio may be 1: 1, 2: 1, 3: 1, 4: 1, 5: 1, 10: 1, 200: 1, 300: 1, 500: 1, 1000: 1 or 5000 :one. An experienced specialist in the art will be able to quickly determine the ratio of water / composition that meets a specific situation or conditions.

In accordance with the present invention, upon dilution of said emulsion pre-concentrate with water, an emulsion is formed having an optical density of more than 0.3 at 400 nm. The optical density of the emulsions formed by diluting the pre concentrates of the emulsions in accordance with the present invention in a ratio of 1: 100 at 400 nm can be in the range, for example, from 0.3 to 4.0. The optical density at 400 nm can be, for example, approximately 0.4, 0.5, 0.6, 1.0, 1.2, 1.6, 2.0, 2.2, 2.4, 2.5 , 3.0 or 4.0. Methods for determining the optical density of liquid solutions are well known to those skilled in the art. An experienced specialist in the art will be able to determine and clarify the relative proportions of the ingredients in the pre-concentrates of the emulsions in accordance with the present invention to obtain, when diluted with water, an emulsion having any specific optical density and within the scope of the present invention.

The pharmaceutical compositions in accordance with the present invention can be, for example, semi-solid preparations or liquid preparations. Semi-solid preparations in accordance with the present invention may be any semi-solid preparations known to those skilled in the art, including, for example, gels, pastes, creams and ointments.

The pharmaceutical compositions of the present invention comprise a lipophilic phase component. Components suitable for use as a component of the lipophilic phase include any pharmaceutically acceptable solvent immiscible with water. Obviously, such solvents have no or practically no surface-active action.

The component of the lipophilic phase may contain mono-, di - or triglycerides. Mono-, di- or triglycerides that can be used in accordance with the present invention include mono-, di- or triglycerides of fatty acids C ₆ , C ₈ , C ₁₀ , C ₁₂ , C ₁₄ , C ₁₆ , C ₁₈ , C ₂₀ and C ₂₂ . Examples of diglycerides, in particular, include dioleic, dipalmitoleic and mixed capryline-caprine diglycerides. Preferred triglycerides include vegetable oils, fish oils, animal fats, hydrogenated vegetable oils, partially hydrogenated vegetable oils, synthetic triglycerides, modified triglycerides, fractionated triglycerides, medium carbon chain triglycerides and long chain triglycerides, structured triglycerides of these compounds also.

Among the above triglycerides, preferred triglycerides include almond oil, babasu oil, borago oil, blackcurrant seed oil, canola oil, castor oil, coconut oil, corn oil, cottonseed oil, evening primrose oil, grape seed oil, grape seed oil, mustard oil oil, olive oil, palm oil, palm kernel oil, peanut oil, rep oil, saffron oil, hemp oil, shark liver oil, soybean oil, sunflower oil, hydrogenated castoro oil, hydrogenated coconut oil, hydrogenated palm oil, hydrogenated soybean oil, hydrogenated vegetable oil, hydrogenated cotton and castor oil, partially hydrogenated soybean oil, partially hydrogenated soybean and cottonseed oil, glyceryl tricaproate, glyceryl triacrylate, glyceryl glycerol, tri-glycerol, , glyceryl trilinolenate, glyceryl tricaprilate / caprate, glyceryl tricaprilate / caprate / laurate, glyceryl tricaprilate / caprate / linoleate and gly ceryltricaprilat / caprate / stearate.

A preferred triglyceride is a medium carbon chain triglyceride sold under the trademark LABKAEAC CC. Other preferred triglycerides include neutral oils, for example, neutral vegetable oils, in particular fractionated coconut oils, such as those known and marketed under the name MUOU, including products: MUOU 810, MUOU 812, MUOU 818 and SARTEX 355.

Caprylic-capric acid triglycerides are also suitable for the purposes of the present invention, such as products known and sold under the brand name MUCHT, including product MUK1TOB 813. Other suitable products of this class are CAPMT MCT, SARTECH

- 5 010437

200, SARTEH 300, SARTEH 800, ΝΕΟΒΕΕ M5 and ΜΑΖΟΕ 1400.

Especially preferred as a component of the lipophilic phase is the product MULO 8 8 (see US patent No. 5342625).

The pharmaceutical compositions of the present invention may also contain a hydrophilic phase component. The hydrophilic phase component may include, for example, a pharmaceutically acceptable C _1-5 -alkyl or tetrahydrofurfuryl diester or an incomplete ether of a low molecular weight mono- or polyoxyalkanediol. Suitable components of the hydrophilic phase include, for example, di- or incomplete, in particular incomplete ethers of mono- or poly, in particular mono- or dioxoalkanediols containing from 2 to 12, in particular 4 carbon atoms. Preferably, the mono- or polyoxyalkanediol moiety is straight chain. Examples of components of the hydrophilic phase used in accordance with the present invention are known and commercially available substances under the trademarks TKAOSITO and SUSUSORIKOE (see US patent No. 5342625).

In a particularly preferred embodiment, the hydrophilic phase component comprises 1,2 propylene glycol.

Of course, the component of the hydrophilic phase in accordance with the present invention may also contain one or more additional ingredients. However, it is preferable that any additional ingredients contain materials in which the active vitamin соединение compound is sufficiently soluble so that the effectiveness of the hydrophilic phase as a carrier medium of the active vitamin соединения compound is not significantly reduced. Examples of possible additional components of the hydrophilic phase include lower (e.g., C _1-5 ) alkanols, in particular ethanol.

The pharmaceutical compositions in accordance with the present invention also contain one or more surfactants. Surfactants that can be used in connection with the present invention include hydrophilic or lipophilic surfactants, or mixtures thereof. Particularly preferred are nonionic hydrophilic and nonionic lipophilic surfactants.

Suitable hydrophilic surfactants include reaction products of natural or hydrogenated vegetable oils and ethylene glycol, i.e. polyoxyethylene glycolated natural or hydrogenated vegetable oils, for example polyoxyethylene glycolated natural or hydrogenated castor oils. Such substances can be obtained by known methods, for example, by reacting natural or hydrogenated castor oil or its fractions with ethylene oxide, for example, in a molar ratio ranging from 1:35 to 1660, with the possible removal of free polyethylene glycol components from the product, for example, in accordance with the methods described in German accepted patent applications No. 1182388 and 1518819.

Suitable hydrophilic surfactants suitable for use in accordance with the present invention also include polyoxyethylene sorbitol fatty acid ester, for example, lauric mono- and triester, palmitic, stearic and oleic acid esters, for example of the types known and commercially available under the brand name ΤνΕΕΕΝ including products:

ΤνΕΕΝ 20 (polyoxyethylene (20) sorbitan monolaurate),

ΤνΕΕΝ 40 (polyoxyethylene (20) sorbitan monopalmitate),

ΤνΕΕΝ 60 (polyoxyethylene (20) sorbitan monostearate),

ΤνΕΕΝ 80 (polyoxyethylene (20) sorbitan monooleate),

ΤνΕΕΝ 65 (polyoxyethylene (20) sorbitan tristearate),

ΤνΕΕΝ 85 (polyoxyethylene (20) sorbitan trioleate),

ΤνΕΕΝ 21 (polyoxyethylene (4) sorbitan monolaurate),

ΤνΕΕΝ 61 (polyoxyethylene (4) sorbitan monostearate) and

ΤνΕΕΝ 81 (polyoxyethylene (5) sorbitan monooleate).

The most preferred products of this class suitable for use in accordance with the present invention are the above products ΤνΤ 40 and ΤνΕΕΝ 80 (see Naieg, e! A1., US patent No. 5342625).

Suitable hydrophilic surfactants for use in pharmaceutical compounds of the invention are also polyoxyethylene esters of alkyl alcohols, polyoxyethylene glycol and fatty acid esters, for example polyoxyethylene glycol and stearic acid esters, polyglycerol and fatty acid esters, polyoxyethylene glycerides, polyoxyethylene glycerides vegetable oils, polyoxyethylene esters of hydrogenated vegetable oils, reaction mixtures of polyols and, for example, fatty acids you, glycerides, vegetable oils, hydrogenated vegetable oils and sterols, copolymers of polyoxyethylene and polyoxypropylene, block copolymers of polyoxyethylene and polyoxypropylene, dioctyl succinate, dioctyl sodium sulfosuccinate, di- [2-ethylhexyl] succinate or sodium lauryl sulfates such as phosphate, for example, soybean lecithins, propylene glycol mono- and diesters and fatty acids, such as, for example, propylene glycol dicaprylate, propylene glycol di laurate, propylene glycol

6 010437 hydroxystearate, propylene glycol isostearate, propylene glycol laurate, propylene glycol ricinoleate, propylene glycol stearate, and especially preferred are propylene glycol diester and capryl capric acid and bile salts, for example, alkali metal salts, in particular sodium taurochol.

Suitable lipophilic surfactants include alcohols, polyoxyethylene ethers of alkyl alcohols, fatty acids, bile acids, esters of glycerol and fatty acids, esters of acetylated glycerol and fatty acids, esters of lower alcohols and fatty acids, polyoxyethylene glycol ethers of glycerol and fatty acids, polyoxyethylene glycol ethers fatty acids, polyoxyethylene glycerides, lactic acid esters of mono / diglycerides, sorbitol fatty acid esters, polyoxyethylene sorbitol fatty acid esters, block copolymers of lyoxyethylene and polyoxypropylene, trans-esterified vegetable oils, sterols, sugar esters, sugar ethers, sugar glycerides, vegetable oil polyoxyethylene ethers, hydrogenated vegetable oil polyoxyethylene ethers, polyol reaction mixtures and at least one member of the group consisting of fatty acids, glycerides , vegetable oils, hydrogenated vegetable oils, sterols, as well as mixtures of these substances.

Lipophilic surfactants suitable for use in pharmaceutical compounds of the present invention also include trans-esterification products of triglycerides from natural vegetable oils and polyalkylene polyols. Such transesterified products are known in the art and can be prepared, for example, in accordance with the general procedures described in US Pat. No. 3,288,824. These include transesterification products of various natural (eg, non-hydrogenated) vegetable oils, for example corn oil, stone seed oil , almond oil, peanut oil, olive oil and palm oil, and mixtures thereof with polyethylene glycols, in particular polyethylene glycols having an average molecular weight of from 200 to 800. Preferred vlyayutsya products obtainable by transesterification of 2 molar parts of natural vegetable oil triglyceride with 1 part of polyethylene glycol (e.g. having an average molecular weight of from 200 to 800). Various forms of transesterification products of a certain class are known and are commercially available under the trade name LABCAN.

Other lipophilic surfactants suitable for use in pharmaceutical formulations of the present invention include oil-soluble derivatives of vitamins, for example, tocopherol PEC-1000 succinate (1eor11cgo 1 PEC-1000 KisschGe) (Vitamin E TPC8).

Suitable lipophilic surfactants for use in pharmaceutical compounds of the present invention are also mono-, di- and mono / diglycerides, in particular esterification products of caprylic or capric acid with glycerol, sorbitol fatty acid esters, esters pentaerythritol and fatty acids and polyalkylene glycol ethers, for example, pentaerythritol-dioleate, distearate, monolaurate, polyglycol ether and monostearate, as well as pentaerythritol and fatty acid esters, mon glycerides, for example glycerol monooleate, glycerol monopalmitate and glycerol monostearate, glycerol triacetate or (1,2,3) -triacetin, and sterols and their derivatives, for example cholesterols and their derivatives, in particular phytosterols, for example products including sitosterol, campesterol or stigma , and their adducts with ethylene oxide, for example soy sterols and their derivatives.

Those skilled in the art will recognize that some of the surfactants supplied contain small to moderate triglycerides, usually as a result of incomplete conversion of the triglyceride starting material, for example, in a transesterification reaction. Thus, surfactants suitable for use in pharmaceutical compositions of the present invention include those surfactants that contain triglycerides. Examples of commercially available triglyceride-containing surfactants include some of the compounds of the following series of surfactants: BEISGKEZ, ΜΑΙ8ΙΝΕ8 and ΙΜ \ νΐΤΟΚ8. Specific examples of such compounds are SES'CHKE 44/14 (saturated glycerides poliglikolizovannye) SES'CHKE 50/13 (saturated glycerides poliglikolizovannye) SEISGKE 53/10 (saturated glycerides poliglikolizovannye) SEISGKE 33/01 (semi-synthetic triglycerides of _C8 -C1 ₈ saturated fatty acids), SEISGKE 39/01 (semi-synthetic glycerides), other SEISGKE, such as 37/06, 43/01, 35 / 10,37 / 02, 46 / 07,48 / 09, 50/02, 62 / 05 etc., MA18SHE 35-1 (linoleic acid glycerides) and ΙΜνίΤΟΚ 742 (caprylic / capric acid glycerides) (see US Patent No. 6267985).

Other supplied surfactant compositions having a significant triglyceride content are known to those skilled in the art. It should be noted that compositions that simultaneously contain both triglycerides and surfactants can serve as all or part of the component of the lipophilic phase in accordance with the present invention, as well as as all or part of surfactants.

Pharmaceutical formulations in accordance with the present invention also include a compound

- 7 010437 active vitamin Ό. The term active vitamin соединение compound, as used herein, means vitamin Ό hydroxylated at least at carbon-1 of cycle A, for example, 1hydroxyvitamin Ό ₃ . For the composition of the present invention, a preferred active vitamin Ό compound is 1a, 25-hydroxyvitamin Ό ₃ , also known as calcitriol. A large number of other active vitamin соединений compounds are known, and they can be used in the practice of the present invention. Examples include Ια-hydroxy derivatives with a side chain length of 17 greater than the length of the side chains of cholesterol or ergosterol (see US patent No. 4717721), cyclopentane analogues of vitamin Ό (see US patent No. 4851401), analogues of vitamin Ό ₃ with alkynyl, alkenyl and alkanyl side chains (see US Pat. Nos. 4,866,048 and 5,145,846), trihydroxycalciferol (see US Pat. No. 5,120,722), fluorocholecalciferol compounds (see US Pat. No. 5,547,947), methyl substituted vitamin Ό (see US Pat. No. 5,446,035), 23-oxa derivatives ( see US patent No. 5411949), compounds of 19-nor-vitamin Ό (see US patent No. 5237110) and hydroxylated derivatives of 24-homo-vitamin Ό (see US patent No. 4857518). Specific examples include KOSLTKOE (Cosquer aogaFpek) suitable for injectable calcitriol SLSIEH explored medicines production Leo RkattaseiksaI including EB 1089 (24a, 26a, 27atrigomo-22,24-diene-1AA, 25- (OH) ₂ -O _3, KH 1060 (20-epi-22-oxa-24a, 26a, 27a-trigomo-1a, 25- (OH) ₂ -E ₃ ), 8eosa1sko1, MS 1288 (1.25- (OH) ₂ -20-epi-E ₃ ) and MS 903 (calcipotriol, 1a, 24k- (OH) ₂ -22-en-26.27 dehydro-E ₃ ), medicines from Koske Rkattaseijs1, which include 1.25- (OH) ₂ -16-eN-E ₃ , 1.25- (OH) ₂ 16-еН-23-in- ₃ and 25- (OH) ₂ -16-en-23-in-О ₃ , Skida1 Rkagtaseiksa And 22-oxacalcitriol (22-oxa-1a , 25 (OH) ₂ -E _3, 1a - (OH) -E ₅ of Illinois university iteta and medicaments of 1ke 1i81yi1e th Mey1sa1 Skeshsch1tu 8sketshd-AD, which include ΖΚ 161422 (20-methyl-1,25- (OH) ₂ -E ₃₎ and ΖΚ 157202 (20-methyl-23-en1,25- (OH ) ₂ -E ₃ ), 1- (OH) -E ₂ , 1- (OH) -O ₃ and 1- (OH) -E _4. Additional examples include 1a, 25- (OH) ₂ 26.27th _6- O ₃ , 1a, 25- (OH) ₂ -2-en-E ₃ , 1a, 25- (OH) _2- P ₃ , 1a, 25- (OH) ₂ -P ₂ , 1a, 25- ( OH) _2- P ₄ , 1a, 24.25- (OH) _3- P ₃ , 1a, 24.25- (OH) _3- O ₂ , 1a, 24.25- (OH) ₃ -P ₄ , 1a - (OH) -25-GO ₃ , 1- (OH) -25-GO ₄ , 1- (OH) -25-GO ₂ , 1a, 24- (OH) ₂ Ό ₄ , 1a, 24- (OH) _2- E ₃ , 1a, 24- (OH) _2- P ₂ , 1a, 24- (OH) ₂ -25-EP ₄ , 1a, 24- (OH) ₂ -25-EP ₃ , 1a, 24- ( OH) ₂ -25-EP ₂ , 1a, 25 (OH) ₂ -26.27-E ₆ -22-en-O ₃ , 1a, 25- (OH) ₂ -26.27-E ₆ -O ₃ , 1a, 258- (OH) ₂ -26-E ₃ -P ₃ , 1a, 25- (OH) ₂ -24-E ₂ -P ₃ ,

1a, 258.26- (OH) ₂ -22-en-O ₃ , 1a, 25K, 26- (OH) ₂ -22-en-P ₃ , 1a, 25- (OH) ₂ -P ₂ , 1a, 25- (OH) ₂ -24-epi-P ₃ , 1a, 25 (OH) ₂ -23-in-O ₃ , 1a, 25- (OH) ₂ -24K-E-O ₃ , 1a, 258.26 - (OH) _2- P ₃ , 1a, 24K- (OH) ₂ -25E-P ₃ , 1a, 25- (OH) ₂ -26,27-E ₆ 23-in-O ₃ , 1a, 25K- ( OH) ₂ -26-E ₃ -O ₃ , 1a, 25.28- (OH) _3- P ₂ , 1a, 25- (OH) ₂ -16-en-23-in-P ₃ , 1a, 24K, 25- (OH) _3- P ₃ , 1a, 25- (OH) ₂ -26.27-E ₆ -23-en-O ₃ , 1a, 25K- (OH) ₂ -22-en-26-E ₃ -P ₃ , 1a, 258- (OH) ₂ -22-en-26-E ₃ -P ₃ , 1a, 25K- (OH) ₂ Ό ₃ -26.26.26-ά ₃ , 1a, 258- ( OH) ₂ -O ₃ minutes _-26,26,26-3 and 1a, 25K- (OH) _{2 -22-ene-3} -26,26,26 About ₃ minutes. Additional examples can be found in XVO 99/49870. See also, for example, US patents No. 5457217, 5447924, 5446034, 5414098, 5403940, 5384313, 5374629, 5373004, 5371249, 5430196, 5260290, 5393749, 5395830,

5250523, 5247104, 5397775, 5194431, 5281731, 5254538, 5232836, 5185150, 5321018, 5086191, 5036061,

5030772, 5246925, 4973584, 5354744, 4927815, 4804502, 4857518, 4851401, 4851400, 4847012, 4755329,

4940700, 4619920, 4594192, 4588716, 4564474, 4552698, 4588528, 4719204, 4719205, 4689180, 4505906,

4769181, 4502991, 4481198, 4448726, 4448721, 4428946, 4411833, 4367177, 4336193, 4360472, 4360471,

4307231, 4307025, 4358406, 4305880, 4279826 and 4248791.

In a preferred embodiment of the present invention, the active vitamin соединение compound has a reduced hypercalcemic effect compared to vitamin Ό, therefore, increased doses of the compound can be administered without causing hypercalcemia in the animal. A reduced hypercalcemic effect is defined as an effect less than the hypercalcemic effect, which causes the introduction of an equal dose of 1a, 25-hydroxyvitamin Ό ₃ (calcitriol). So, for example, the hypercalcemic effect of EB 1089 is equal to 50% of the hypercalcemic effect of calcitriol. Additional active vitamin соединения compounds having a reduced hypercalcemic effect include Ko23-7553 and Ko24-5531 manufactured by Neutap LaKoske. Other examples of active vitamin соединений compounds having a reduced hypercalcemic effect are described in US Pat. No. 4,771,721.

Methods for determining the hypercalcemic effect of active vitamin соединений compounds are known to those skilled in the art and are described in the literature, for example, Napkep e1 a1., Sigg. Rkagt. Eh5. 6: 803-828 (2000).

The pharmaceutical compositions in accordance with the present invention may also contain one or more additives. Supplements known in the art include, for example, tackifiers, antifoam agents, buffering agents, antioxidants (e.g. ascorbyl palmitate, butyl hydroxyanisole (BHA), butyl hydroxytoluene (BHT) and tocopherols, for example, atocoferol (vitamin E)), preservatives, chelating agents, viscosity modulators, toning agents, flavoring agents, colorants, perfumes, silencers, suspending agents, binders, fillers, plasticizers, lubricants, and also mixtures indicated substances. One skilled in the art can easily determine the content of such additives necessary to obtain the desired properties. Thus, for example, antioxidants may be present in an amount of about 0.01 to 0.5% by weight of the total composition, preferably about 0.05 to 0.35%.

- 8 010437

The additive may also contain a thickening agent. Suitable thickeners can be thickeners known and used in the art, including, for example, pharmaceutically acceptable polymeric materials and inorganic thickeners. Examples of thickeners suitable for use in pharmaceutical formulations of the present invention include polyacrylate resins and resins of copolymers of polyacrylates, for example resins of polyacrylic acid and copolymer resins of polyacrylic acid / methacrylic acid, cellulose and cellulose derivatives including alkyl celluloses, for example methyl, ethyl, and propyl cellulose, hydroxyalkyl cellulose, for example hydroxypropyl cellulose and hydroxypropylalkyl cellulose, such as hydroxypropyl methyl cellulose, acylated c celluloses, for example cellulose acetates, cellulose acetate phthalates, cellulose acetate succinates and hydroxypropyl methyl cellulose phthalates, and salts of these compounds, such as sodium carboxymethyl cellulose, including polyvinylpyrrolidones, for example, polyvinylpyrrolidones, vinyl vinylpyrrolinyl copolymers, vinylpyrrolinyl vinylpyrrolinyl vinyl alcohols, as well as other polymeric materials, including tragacanth gum, gum arabic, alginates, for example alginic acid, and Whether these compounds, for example sodium alginate, and inorganic thickeners such as atapulgit, bentonite and silicates including hydrophilic silicon dioxide products, e.g. alkylated (for example methylated) silica gels, in particular colloidal silicon dioxide products.

Such thickeners, as described above, may be included, for example, to create a sustained release effect. However, if the medication is intended for oral administration, the use of thickeners, as described above, is generally not required and, as a rule, less preferred. On the other hand, the use of a thickener is indicated if the medication is intended, for example, for topical use.

The compositions in accordance with the present invention can be used in any suitable way, for example, orally, in particular in unit dosage form, for example, in solution, in soft or hard capsule form, including gelatin capsule form, for example parenteral or topical, in particular for application to the skin, for example, in the form of a cream, paste, lotion, gel, ointment, lotion, poultices, patch, skin stickers, or the like, or for ophthalmic use, for example, the form of eye drops, eye lotions or eye gels. Lightly flowing forms can also be used, for example, solutions or emulsions, in particular for administration to the lesion, or they can be administered perrectally, i.e. in the form of an enema. The compositions may further contain agents that increase the delivery of the active vitamin соединения compound, for example liposomes, polymers or copolymers (in particular branched chain polymers).

If the composition according to the present invention is prepared in unit dosage form, the active vitamin соединение compound is preferably present in an amount of 1 to 400 μg per unit dose. More preferably, the content of the active vitamin соединения compound in a standard dose is about 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25, 30, 35, 40, 45, 50, 55 , 60, 65, 70, 75, 80, 85, 90, 95, 100, 105, 110, 115, 120, 125, 130, 135, 140, 145, 150, 155, 160, 165, 170, 175, 180 , 185, 190, 195, 200, 205, 210, 215, 220, 225, 230, 235, 240, 245, 250, 255, 260, 265, 270, 275, 280, 285, 290, 295, 300, 305 , 310, 315, 320, 325, 330, 335, 340, 345, 350, 355, 360, 365, 370, 375, 380, 385, 390, 395 or 400 mcg or any other value within these limits. In a preferred embodiment, the content of the active vitamin соединения compound in a unit dose is from about 5 to 180 μg, more preferably from about 10 to 135 μg, even more preferably about 45 μg. In one embodiment, the unit dosage form contains 45, 90, 135, or 180 μg of calcitriol.

If the unit dosage form of the composition is a capsule, the total amount of ingredients present in the capsule is preferably about 10-1000 μl. More preferably, the total amount of ingredients present in the capsule is approximately 100-300 μl. In another embodiment, the total amount of ingredients present in the capsule is preferably about 10-1500 mg, preferably about 1001000 mg. In one embodiment, the total amount is about 225, 450, 675, or 900 mg. In another embodiment, the unit dosage form is a capsule containing 45, 90, 135 or 180 μg of calcitriol.

The relative proportions of the ingredients in the composition in accordance with the present invention, of course, should substantially depend on the particular composition. The relative proportions can also greatly depend on the specific function of the ingredients of the composition. The relative proportions can also be changed depending on the specific ingredients used and the desired physical characteristics of the final composition, for example, if the composition is intended for topical use, depending on whether it is a free-flowing liquid or paste. An average person skilled in the art will be able to determine the working proportions of the components in each specific example. All proportions and relative ranges of weights below should be understood.

- 9 010437 only as preferred or individually designed guidelines, and not as limitations of the invention in its broadest aspect.

The component of the lipophilic phase in accordance with the present invention may suitably be present in amounts of from about 30 to 90 wt.% Of the total weight of the composition. Preferably, the component of the lipophilic phase is present in amounts of from about 15 to 65 wt.% Of the total weight of the composition.

The surfactant or surfactants in accordance with the present invention may be present in amounts of from about 1 to 90 wt.% Of the total weight of the composition. Preferably, the surfactant or surfactants are present in amounts of from about 5 to 85 wt.% Of the total weight of the composition.

The amount of the active vitamin соединения compound in the compositions of the present invention, of course, can be changed, for example, depending on the intended route of administration and on the percentage of the remaining components. However, in the General case, the compound of active vitamin Ό in accordance with the present invention may be present in an amount of from about 0.005 to 20 wt.% Of the total weight of the composition. Preferably, the active vitamin соединение compound is present in an amount of about 0.01 to 15% by weight of the total composition.

The hydrophilic phase component in accordance with the present invention may be present in amounts of from about 2 to 20 wt.% Of the total weight of the composition. Preferably, the hydrophilic phase component is present in amounts of from about 5 to 15 wt.% Of the total weight of the composition.

The pharmaceutical composition in accordance with the present invention may be a semi-solid dosage form. Semi-solid dosage forms in accordance with the present invention may include, for example, from about 50 to 80 wt.% Component of the lipophilic phase of the total weight of the composition, from about 5 to 35 wt.% Surfactant of the total weight of the composition and from about 0, 01 to 15 wt.% Compounds of active vitamin Ό of the total weight of the composition.

The pharmaceutical compositions of the present invention may be in liquid dosage form. Liquid dosage forms in accordance with the present invention may contain, for example, from about 50 to 60 wt.% Component of the lipophilic phase of the total weight of the composition, from about 4 to 25 wt.% Surfactant of the total weight of the composition, from about 0, 01 to 15% by weight of the active vitamin соединения compound, based on the total weight of the composition, and from about 5 to 10% by weight of the hydrophilic phase component of the total weight of the composition.

In one embodiment of the invention, the pharmaceutical compositions comprise a compound of active vitamin Ό, a lipophilic component, and a surfactant. The lipophilic component may have any percentage from about 1 to 100%. The content of the lipophilic component may be approximately 1,2,3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99 or 100%. The surfactant may have any percentage from about 1 to 100%. The content of surfactant may be approximately 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12,

13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42,

43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72,

73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99 or 100%.

In one embodiment, the lipophilic component is MUOUR 812, and the surfactant is vitamin Ε TPO8. In preferred embodiments, the pharmaceutical formulations contain about 50% MUYOU 812 and about 50% vitamin Ε TPO8, about 90% MUYOU 812 and about 10% vitamin Ε TPO8 or about 95% MUYOU 812 and about 5% vitamin РО TPO8.

In another embodiment of the invention, the pharmaceutical compositions comprise an active vitamin витами compound and a lipophilic component, for example, approximately 100% MULO 812.

In a preferred embodiment of the invention, the pharmaceutical formulations contain approximately 50% MULO 812, approximately 50% Vitamin РО TPO8 and small amounts of BHA and BHT. As shown below, these compositions are extremely resistant, both chemical and physical (see Example 16). In a particularly preferred embodiment of the invention, the pharmaceutical compositions contain approximately 50% MULOUM 812, approximately 50% vitamin РО TPO8 and approximately 0.35% BHA and BHT. Increased durability provides a longer shelf life of compounds. It is important to note that the stability of the compounds allows them to be stored at room temperature, eliminating the difficulties and costs associated with storage during cooling. In addition, such a compound is suitable for oral administration and, as shown below, is capable of dissolving high doses of the active vitamin соединения compound, providing the administration of a high pulse dosage active vitamin соединения compound for the treatment of hyperproliferative diseases and other disorders.

- 10 010437

In addition to the foregoing, the present invention also provides a method for preparing the aforementioned pharmaceutical composition, the method comprising obtaining a homogeneous mixture of individual components and, if necessary, introducing the resulting composition into a standard dosage form, for example, filling said composition with gelatin capsules, in particular from soft or hard gelatin, or filling capsules made from another material.

In a more specific embodiment, the present invention also provides a method for preparing a pharmaceutical composition, said method comprising preparing a homogeneous mixture of an active vitamin соединения compound, for example, calcitriol, as well as a component of the lipophilic phase and a surfactant, as described above, with relative proportions of the component of the lipophilic phase and the surfactant is selected depending on the amount of the active vitamin соединения compound used, thereby obtaining emulsion concentrate.

The present invention also provides methods of treating and preventing hyperproliferative diseases, in particular cancer and psoriasis, said methods comprising administering an active vitamin соединения compound in the form of an emulsion preconcentrate preparation to a patient who needs it. Alternatively, the patient may take the active vitamin соединение compound in the form of an emulsion preparation prepared by diluting with an appropriate amount of water an emulsion preconcentrate according to the present invention. Alternatively, the active vitamin соединение compound may be administered in any composition described herein.

The term cancer, as used herein, refers to all known cancers and can include, but is not limited to, the following diseases: leukemia, in particular acute leukemia, acute lymphocytic leukemia, acute myelocytic leukemia, including myeloid, promyelocytic, myelomonocytic monocytic and erythroleukemic leukemia and myelodysplastic syndrome; chronic leukemia, in particular chronic myelocytic (granulocytic) leukemia, chronic lymphocytic leukemia, fleecy cell leukemia; true polycythemia; lymphomas, in particular Hodgkin's disease and other lymphophmatoses; multiple myelomas, in particular sluggish multiple myeloma, noncreative myeloma, osteosclerotic myeloma, plasma cell leukemia, solitary plasmacytoma and extramedullary plasmacytoma; Waldenstrom macroglobulinemia; monoclonal gammopathy of unknown etiology; benign monoclonal gammopathy; heavy chain disease; sarcomas of bone and connective tissues, in particular bone sarcoma, osteosarcoma, chondrosarcoma, Evaring sarcoma, malignant tumor of giant cells, bone fibrosarcoma, chordoma, parostal sarcoma, soft tissue sarcoma, angiosarcoma lymphoma, sarcoma leukarosarcoma, fibrosarcoma , neurolemma, rhabdomyosarcoma and synovial sarcoma; brain tumors, in particular glioma, astrocytoma, glioma of the brain stem, ependymoma, oligodendroglioma, nonglial tumor, auditory nerve neurilema, craniopharyngioma, medulloblastoma, meningioma, pineocytoma, pineoblastoma and primary brain lymphoma; breast cancer, in particular adenocarcinoma, lobular (small cell) carcinoma, comedocarcinoma, medullary breast cancer, mucoid breast cancer, tubular breast cancer, papillary breast cancer, breast nipple cancer (Paget's disease), inflammatory breast cancer ; cancer of the adrenal gland, in particular pheochromocytoma and adrenocortical carcinoma; thyroid cancers, in particular papillary or follicular thyroid cancer, medullary thyroid cancer and anaplastic thyroid cancer; cancers of the pancreas, in particular insulinoma, gastrinoma, glucagon, vipoma, a tumor releasing somatostatin, and a carcinoid tumor or insulinoma; pituitary cancers, in particular a tumor secreting prolactin and acromegaly; eye cancers, in particular ocular melanoma, iris melanoma, choroid melanoma, ciliary body melanoma and retinoblastoma; vaginal cancers, in particular squamous cell carcinoma, adenocarcinoma and melanoma; cancers of the vulva, in particular squamous cell carcinoma, melanoma and adenocarcinoma, basal cell carcinoma, sarcoma and Pagetova genital disease; cancers of the cervix, in particular squamous cell carcinoma and adenocarcinoma, cancers of the uterus, in particular endometrial carcinoma and uterine cancer; cancers of the ovary, in particular epithelial carcinoma of the ovary; epithelial border tumor of the ovary, germinoma and stromal tumor; cancer of the esophagus, in particular squamous cell carcinoma, adenocarcinoma, adenoid cystic carcinoma, mucus-forming squamous cell carcinoma, adeno-squamous cell carcinoma, sarcoma, melanoma, plasmacytoma, warty carcinoma and oat cell carcinoma; cancers of the stomach, in particular adenocarcinoma, mushroom (polypoid) ulcerative superficial and diffuse spread, malignant lymphoma, liposarcoma, fibrosarcoma and carcinosarcoma; colon cancers; colorectal cancers; liver cancers, in particular liver epithelioma and hepatoblastoma; cancer of the gallbladder, in particular adenocarcinoma; cholangiocarcinomas, in particular papillary, nodular and diffuse; lung cancer, in particular non-small cell lung cancer, squamous cell carcinoma (epidermoid cancer), adenocarcinoma, large cell carcinoma and small

- 11 010437 cell lung cancer; testicular cancer, in particular a terminal tumor, seminoma, anaplastic classical (typical) spermatocytic nonsiminoma, embryonic carcinoma, teratoid carcinoma and choriocarcinoma (yolk sac tumor); prostate cancers, in particular adenocarcinoma, leiomyosarcoma, rhabdomyosarcoma; penile cancer; cancers of the oral cavity, in particular squamous cell carcinoma; basal cell carcinoma; cancers of the salivary gland, in particular adenocarcinoma, mucoepidermoid carcinoma, adenoid cystic carcinoma; cancers of the pharynx, in particular squamous cell carcinoma, warty carcinoma; skin cancers, in particular basal cell carcinoma, squamous cell carcinoma and melanoma, superficially spreading melanoma, nodular melanoma, malignant lentigo melanoma, acral lentigenic melanoma; cancers of the head and neck; renal cancer, in particular renal cell carcinoma, adenocarcinoma, hypernephroma, fibrosarcoma, transitional cell carcinoma (renal pelvis and / or ureter); adenomyosarcoma; and cancers of the bladder, in particular transitional cell carcinoma, squamous cell carcinoma, adenocarcinoma and carcinosarcoma. In addition, cancers that can be treated by the methods and compositions of the present invention include myxosarcoma, osteoblastosarcoma, endotheliosarcoma, lymphangioendotheliosarcoma, mesothelioma, synovioma, hemangioblastoma, epithelial carcinoma, cystadenocarcinoma, bronchogenic carcinoma, carcinoma of the carcinoma, and carcinoma of the carcinoma papillary adenocarcinoma. An overview of such diseases is given in the work of Rshchtap s1 a1., 1985, Mebushe, 2 ' ¹ Eb., TV. Irršsoi So., Rl1abe1rYa, RA aib Migryu e1 a1., 1997, 1pRogteb YesShopk: Thieu Sotr1e1e Vook οί Sapseg Οίηβηοδίδ. TgeaTspeSh apb Vesouegu, U1ktd Reps, Ute Wogk, ΝΥ.

The active vitamin Ό compound is usually administered at a dose of from about 1 to 400 μg, preferably from about 15 to 300 μg. In one specific embodiment, an effective amount of an active vitamin соединения compound is 3, 4, 5, 10, 20, 25, 30, 35, 40, 45, 50, 55, 60, 65, 70, 75, 80, 85, 90 , 95, 100, 105, 110, 115, 120, 125, 130, 135, 140, 145, 150, 155, 160, 165, 170, 175,

180, 185, 190, 195, 200, 205, 210, 215, 220, 225, 230, 235, 240, 245, 250, 255, 260, 265, 270, 275, 280, 285,

290, 295, 300, 305, 310, 315, 320, 325, 330, 335, 340, 345, 350, 355, 360, 365, 370, 375, 380, 385, 390, 395,

400 mcg or more. In some embodiments, the dose of the active vitamin соединения compound is from about 1 to 270 μg, preferably from about 15 to 300 μg, more preferably from about 15 to 225 μg, more preferably from about 15 to 180 μg, more preferably from about 15 to 135 μg, more preferably from about 20 to 90 μg, more preferably from about 30 to 60 μg, and even more preferably about 45 μg. In some embodiments, the methods of the present invention comprise administering an active vitamin соединения compound at a dose of about 0.12 to 3 μg / kg body weight. The compound can be administered in one of the ways, including oral, intramuscular, intravenous, parenteral, perrectal, nasal, topical or transdermal.

In the case of daily administration of the compound, the dose can be kept low, for example about 0.5-5 μg, to exclude or reduce the induction of hypercalcemia. If the active vitamin соединение compound has a reduced hypercalcemic effect, a higher daily dose can be administered without causing hypercalcemia, for example, about 10-20 μg or more (up to about 50-100 μg).

In a preferred embodiment of the invention, the active vitamin соединение compound is administered in a pulsed dosage regimen that allows high doses of the active vitamin соединения compound to be administered without causing hypercalcemia. A pulsed dosage is called intermittent administration of an active vitamin соединения compound in accordance with a continuous intermittent administration or with an intermittent intermittent administration. High doses of the active vitamin соединения compound include doses exceeding approximately 3 μg, as described in the sections above. Thus, in some embodiments of the present invention, methods of treating or reducing the intensity of cancer include pulsed administration of high doses of active vitamin Ό compounds. The frequency of the pulsed dosage can be limited by a number of factors, including, but not limited to, the pharmacokinetic parameters of the compound or composition and the pharmacodynamic effect of the active vitamin соединения compound on the animal. For example, in animals with cancer and having impaired renal function, less frequent administration of the active vitamin соединения compound may be required due to the reduced ability of their body to release calcium.

The following examples serve only to illustrate that the term pulsed dosing can include any intermittent regimen defined by a person skilled in the art.

In one example, the active vitamin соединение compound can be taken no more than once every three days, four days, five days, six days, seven days, eight days, nine days, ten days, ten days , two weeks, three weeks, four weeks, five weeks, six weeks, seven weeks, eight weeks or less. Reception may last one, two, three or four weeks or one, two, three months or longer. After a break, taking the active vitamin соединения compound

- 12 010437 can be repeated in the same or a different application pattern. A break can be one, two, three, four weeks or more, depending on the pharmacodynamic effects that the compound of active vitamin Ό has on the animal.

In another example, the active vitamin соединение compound can be taken once a week for three months.

In one preferred embodiment, the active vitamin соединение compound can be taken once a week for a cycle of three or four weeks. After a break of one week, the intake of the active vitamin соединения compound can be repeated according to the same or different application regimen.

In another preferred embodiment, the active vitamin соединение compound can be taken once every three weeks.

Other examples of application patterns that can be used in the methods of the present invention are described in US Pat. No. 6,521,608, incorporated herein by reference in its entirety.

The above examples of application patterns are for illustrative purposes only and should not be construed as limitations. It will be apparent to those skilled in the art that all active vitamin соединения compounds are included in the scope of the invention and that the exact dosage and regimen of active vitamin соединений compounds can vary depending on many factors.

The magnitude of a therapeutically effective dose of a pharmaceutical agent in the emergency or long-term treatment of a disease or disorder may differ depending on factors including, but not limited to, the nature of the disease or disorder to be treated, specific pharmaceutical agents and the method of administration. In accordance with the methods of the present invention, an effective dose of an active vitamin соединения compound is a dose of the compound effective to treat or reduce the intensity of cancer or other hyperproliferative diseases. A high dose of the active vitamin соединения compound may be a dose in the range of about 3 to 400 μg, or some dose in this range, as described above. The size of the dose, frequency of administration, duration of administration, or a combination thereof may also vary depending on the age, body weight, reaction and medical history of the animal, as well as on the method of administration, pharmacokinetics and pharmacodynamic effects of pharmaceutical agents. Specialists in the art take these factors into account in the established manner.

The rate of absorption and release of compounds of active vitamin Ό is influenced by many factors that are well known to specialists in this field of technology. As indicated above, the pharmacokinetic properties of active vitamin Ό compounds limit the peak concentration of vitamin соединений compounds that can be obtained in the blood without causing hypercalcemia. The rate and extent of absorption, distribution, binding, or localization in tissues, biotransformation, and excretion of an active vitamin соединения compound may affect the frequency of administration of pharmaceutical agents. In some embodiments, active vitamin соединения compounds are administered in high dose pulsed dosage regimen for treating or decreasing cancer intensity in accordance with the above application regimen.

In one embodiment, the active vitamin соединение compound is taken at a dose sufficient to provide a peak plasma concentration of the active vitamin соединения compound in plasma of about 0.1 to 20 nM. In some embodiments, the methods of the present invention comprise administering an active vitamin соединения compound with a dose that provides peak plasma concentrations of 0.1, 0.2, 0.3, 0.4, 0.5, 0.6 , 0.7, 0.8, 0.9, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 12.5, 15, 17.5, 20 nM or lying in some or a range of indicated concentrations. In other embodiments, the active vitamin соединение compound is taken at a dose that provides peak plasma concentrations of the active vitamin соединения compound in excess of about 0.5 nM, preferably from about 0.5 to 20 nM, more preferably from about 1 to 10 nM, more preferably from about 1 to 7 nM and even more preferably from about 3 to 5 nM.

In another preferred embodiment, the active vitamin соединение compound is taken with a dose of at least about 0.12 μg / kg body weight, more preferably with a dose of at least about 0.5 μg / kg body weight.

It will be apparent to those skilled in the art that these unit doses are for an adult patient with an average size and weight of approximately 70 kg and can be adjusted according to factors that are considered in the established manner, as described above.

In some embodiments, the methods of the present invention also include administering a dose of the active vitamin быстро compound, which rapidly produces a peak plasma concentration, for example, within 4 hours. In other embodiments, the methods of the present invention also include administering a dose of the active vitamin Ό vitamin Ό, which is quickly eliminated, for example, has a half-life of less than 12 hours.

Getting high concentrations of the active vitamin соединения compound is useful, however,

- 13 010437 it must be balanced with clinical safety, for example with the absence of hypercalcemia. Therefore, in one aspect of the invention, the methods of the invention include pulsed administration of high doses of active vitamin соединений compounds to a subject suffering from cancer or other hyperproliferative disease, and monitoring the absence of symptoms associated with hypercalcemia in said subject. Such symptoms include calcification of soft tissues (e.g., cardiac tissue), increased bone density, and hypercalcemic nephropathy. In a further embodiment, the methods of the invention include pulsed administration of high doses of active vitamin соединений compounds to a subject suffering from cancer or other hyperproliferative disease, and monitoring the plasma concentration of said subject to prevent an excess of plasma calcium concentration of about 10.2 mg / dl.

In some embodiments, high levels of vitamin соединений compounds in the blood can be safely obtained in combination with a decrease in calcium transport to the blood. In one embodiment, high concentrations of the active vitamin соединения compound can be safely obtained without hypercalcemia if taken in combination with a diet that includes low calcium. In one example, calcium can be captured by an adsorbent, absorbent, ligand, chelate, or other binding component that cannot be transported into the bloodstream through the small intestine. In another example, the activation rate of osteoblast can be inhibited by the administration of, for example, bisphosphonate, in particular zoledronate, pamidronate or alendronate, or glucocorticoid, for example prednisone or dexamethasone, in combination with the active vitamin соедин compound.

In some embodiments, high blood levels of active vitamin соединения compounds are safely obtained in combination with maximizing the rate of calcium release. In one example, calcium excretion can be increased by providing adequate hydration and salt intake. In another example, diuretic therapy may be used to increase calcium excretion.

In some embodiments, methods for treating and preventing hyperproliferative diseases, in particular cancer and psoriasis, also include administering a chemotherapeutic agent or radiotherapeutic agent along with the active vitamin Ό compound.

The term chemotherapeutic agent used in the present description, refers to any chemotherapeutic agent known to specialists in this field of technology as an effective tool for treating or reducing the intensity of cancer. Chemotherapeutic agents include, but are not limited to, small molecules, synthetic drugs, peptides, polypeptides, proteins, nucleic acids (e.g., DNA and RNA polynucleotides, including, but not limited to, antisense nucleotide sequences, triple helices and nucleotide sequences encoding biologically active proteins, polypeptides or peptides), antibodies, synthetic or natural inorganic molecules, mimetic agents, and synthetic or natural organic molecules. Any agent that is known to be beneficial, or which has been or is currently being used to treat or reduce the intensity of cancer, can be used in combination with the active vitamin Ό compound of the invention. Information on therapeutic agents currently used to treat or reduce the intensity of cancer is given, for example, in the work of Nagbtap s1 a1., Eb§., 1996, Soobtap &Sbtap'k THe Ryagtasooshchsa1 WaDk O £ Tyegaraibsk 9 ' ¹ ' EB, Ms-Sga ^ -NS, Youe Wagk, ΝΥ.

Chemotherapeutic agents suitable for use in the methods and compositions of the present invention include alkylating agents, antimetabolites, antimitotic agents, epipodophyllotoxins, antibiotics, hormones and hormone antagonists, enzymes, platinum coordinator complexes, anthracenodions, ureas, imrazine derivatives, methyl derivatives , cytoprotective agents, DNA topoisomerase inhibitors, biological reaction modifiers, retinoids, therapeutic antibodies, different ntsiruyuschie agents, immunomodulatory agents, inhibitors of angiogenesis.

Other chemotherapeutic agents that may be used include abarelix, aldesleukin, alemtuzumab, aliretinoin, allopurinol, altretamine, amifostine, anastrozole, arsenic anhydride, asparaginase, BCG live vaccine, bevaceizumab, camparotinus, blepti benecytin, blemecinum, blemeci, blemecinum, blemecine, blemeci , carboplatin, carmustine, celecoxib, cetuximab, chlorambucil, cinacalcet, cisplatin, cladribine, cyclophosphamide, cytarabine, dacarbazine, dactinomycin, darbepoetin alpha, daunorubicin, denileukin diptitox, dexr zoksan, docetaxel, doxorubicin, dromostanolone, solution B Elliott, epirubicin, epoetin alfa, estramustine, etoposide, exemestane, filgrastim, floxuridine, fludarabine, fluorouracil, fulvestrant, gemcitabine, gemtutsumab ozogamicin, gefitinib, goserelin, hydroxyurea, ibritumomab tiuxetan, idarubicin, izofamid , imatinib, interferon alfa-2a, interferon alfa-2b, irinotecan, tetrozole, leucovorin, levamisole, lomustine, mecloretamine, megestrol, melphalan, mercaptopurine, mesna, methotrexate, methoxalen, methylprednisolone, mitotomyne, mitomycin Ntron, Nandrolone, Nofetumomab, Oblimersen, Oprelvekin, Oxaliplatin, Paclitaxel, Pamidronate, Pegademas, Peraspargas, Pegfilgrastim, Pemetrexed, Pentostatin, Pipobroman, Plykamycin, Polypherosancrambrin, Razcarbincrimbasbramcarbazrbincrimbase, Ricarzincrimbasbrycrimbasbrycrimbase, Rascarbincrimbasbrycrimbasbrycrimbasbincarbrambasrincarbazrincarbazrincarbrambasrincarbazrimbasin Tarceva, temozolomide, teniposide, testolactone,

- 14 010437 thioguanine, thiotepa, topotecan, toremifene, tositumomab, trastuzumab, tretinoin, uracil mustard, valrubicin, vinblastine, vincristine, vinorelbine and zoledronate.

Chemotherapeutic agents can be administered in doses known to those skilled in the art as being effective in treating pancreatic cancer. In some embodiments, chemotherapeutic agents can be administered at lower doses than usual due to the addition or synergistic effect of the active vitamin соединения compound.

The term radiotherapeutic agent used in the present description, without limitation, refers to any radiotherapeutic agent known to specialists in this field of technology as an effective tool for treating or reducing the intensity of cancer. So, for example, a radiotherapeutic agent may be one of the agents used in brachytherapy or radioisotope therapy.

In brachytherapy, you can use, without limitation, any application, dose or method known to specialists in this field of technology as effective for treating or reducing the intensity of cancer. In general, brachytherapy involves the introduction of radioactive sources into the body of a subject suffering from a cancer, preferably directly into the tumor, so that the tumor is exposed to the maximum effect of the radioactive source, while minimizing the effect on healthy tissue. Examples of isotopes that can be used in brachytherapy include, but are not limited to, phosphorus 32, cobalt 60, palladium 103, ruthenium 106, iodine 125, cesium 137, iridium 192, xenon 133, radium 226, California 252 and gold 198. Methods of administration, devices and compositions used in brachytherapy are described in the work of Mahsgop e1 a1., 8et. Kai Oops. 12: 95-108 (2002) and in US patents No. 6319189, 6179766, 6168777, 6149889 and 5611767.

In a radioisotope therapy, it is possible to use, without limitation, any application schedule, dose or method known to those skilled in the art as being effective in treating or reducing the intensity of a cancer. In general, radioisotope therapy involves the systematic administration of a radioisotope that accumulates in cancer cells or binds to their surface. The preferred accumulation of the radioisotope can be realized by various mechanisms, including, in particular, but not limited to, the introduction of the radioisotope into rapidly proliferating cells, the specific accumulation of the radioisotope in cancerous tissue without special targeting, or the conjugation of the radioisotope with a neoplasm-specific biomolecule.

Examples of radioisotopes that can be used in radioisotope therapy include, but are not limited to, phosphorus 32, yttrium 90, dysprosium 165, indium 111, strontium 89, samarium 153, rhenium 186, iodine 131, iodine 125, lutetium 177 and bismuth 213 Although all of these isotopes can bind to a biomolecule, providing a specific orientation, iodine 131, indium 111, phosphorus 32, samarium 153 and rhenium 186 can be introduced systematically without this conjugation. One of skill in the art can select a specific biomolecule to provide targeted effects on a particular neoplasm for the purpose of conducting radioisotope therapy using surface molecules of cells present in this neoplasm. Examples of biomolecules providing specificity for specific cells are described in the article Tyotak, Sapseg VuPeg. Kayyuryagt. 17: 71-82 (2002), which is incorporated herein by reference in its entirety. In addition, methods of administration and formulations used in radioisotope therapy are described in US Pat. Nos. 6,264,400, 6,358,194 and 5,766,571.

The term radiotherapeutic treatment used in this description, without limitation, refers to any radiotherapeutic treatment methods known to specialists in this field of technology as effective for treating or reducing the intensity of cancer. So, for example, radiotherapy treatment can be external radiation therapy, thermotherapy, radiosurgery, charged particle radiotherapy, neutron radiotherapy or photodynamic therapy.

In radiation therapy with external radiation, you can use, without limitation, any application, dose or method known to specialists in this field of technology as effective for treating or reducing the intensity of cancer. In the general case, radiotherapy with external radiation involves irradiating a certain volume of the subject's body with high-energy radiation that causes cell death in this volume. The irradiated volume preferably includes the entire area affected by the cancer to be treated, and preferably the minimum possible amount of healthy tissue. Administration methods, devices and compositions used in external radiation therapy are described in US Pat. Nos. 6,449,336, 6398710, 6393096, 6335961, 6307914, 6256591, 6245005, 6038283, 6001054, 5802136, 5596619 and 5528652.

In thermotherapy, you can use, without limitation, any application, dose or method known to specialists in this field of technology as effective for treating or reducing the intensity of cancer. In some embodiments, the thermotherapy may be cryodestruction therapy. In other embodiments, thermotherapy may be hyperthermic therapy. In addition, there are options for implementation in which thermotherapy can be a therapy that increases the temperature of the tumor to a greater extent,

- 15 010437 than hyperthermic therapy.

Cryodestruction therapy involves freezing the neoplastic mass, which leads to the formation of intracellular and extracellular ice crystals, the destruction of cell membranes, proteins and organelles and the creation of a hyperosmotic environment, causing cell death. The methods and devices used in cryodestruction therapy are described in the work of Migra e! A1., 8et. Yoke 1. Opso1. 79: 133-140 (2001) and US Pat. Nos. 6,383,181, 6383180, 5993444.5654279, 5437673 and 5147355.

Hyperthermic therapy usually involves increasing the temperature of the neoplastic mass in the range from about 42 to 44 ° C. The temperature of the cancerous tumor can be increased even above the specified limit, however, this can increase the damage to the surrounding healthy tissue, without leading to an increase in cell mortality inside the tumor to be treated. With hyperthermic therapy, the tumor can be heated without limitation by any means known to those skilled in the art. In particular, but without limitation, the tumor can be heated by microwave radiation, high-intensity focused ultrasound, ferromagnetic thermocouples, localized electric fields, infrared radiation, wet or dry high-frequency ablation, laser photocoagulation, laser interstitial thermotherapy and electroacoustics. Microwave and high-frequency radiation can be generated using waveguide applicators, as well as horn, spiral, sheet and compact applicators.

Other methods, devices and compositions that are used to increase the temperature of the tumor are described in the article HUNAC e! A1., baps! Opso1. 3: 487-97 (2002) and US Pat. Nos. 6,470,217, 6,379,347, 6165440, 6163726, 6099554, 6009351, 5776175, 5707401, 5658234, 5620479, 5549639 and 5523058.

In radiosurgery, you can use, without limitation, any application, dose or method known to specialists in this field of technology as effective for treating or reducing the intensity of cancer. In the general case, radiosurgery includes the action of a manually directed radioactive source on a specific volume of the subject's body, which causes cell death in the specified volume. The irradiated volume preferably contains all of the cancer to be treated and the smallest possible amount of healthy tissue. Typically, the tissue to be treated is first exposed to conventional surgical methods, and then the surgeon manually sends a radioactive source to it. In an alternative embodiment, the radioactive source can be installed near the tissue to be irradiated, for example, using a laparoscope. The methods and devices used in radiosurgery are also described in the paper. A1., Eig. 1. 8igd. Opso1. 28: 180-185 (2002) and in US patents No. 4621416, 6248056 and 5547454.

In radiation therapy using charged particles, you can use, without limitation, any regimen, dose or method known to those skilled in the art as effective in treating or reducing the intensity of a cancer. In some embodiments, the charged particle radiotherapy may be proton beam radiotherapy. In other embodiments, charged particle radiotherapy may be helium-ion radiotherapy. In the general case, radiotherapy using charged particles involves irradiating a certain volume of the subject's body with a beam of charged particles, which causes cell death in the specified volume. The irradiated volume preferably contains all of the cancer to be treated and the smallest possible amount of healthy tissue. A method for realizing radiotherapy using charged particles is described in US Pat. No. 5,668,371.

In neutron radiotherapy, you can use, without limitation, any application, dose or method known to specialists in this field of technology as effective for treating or reducing the intensity of cancer. In some embodiments, charged particle neutron radiotherapy may be neutron capture therapy. In such embodiments, a subject is administered a compound that emits radiation upon neutron bombardment and preferably accumulates in a neoplastic mass. The tumor is then irradiated with a low-energy neutron beam, activating the compound and causing it to emit decay products that kill the tumor cells. The compound to be activated can preferably be accumulated in the target tissue by one of the methods used for radioisotopes, as described above, or by the methods described in Lagotogue, 8 ett. Opso1. 24: 672-685 (1997) and in US patents No. 6400796,5877165, 5872107 and 5653957.

In other embodiments, neutron radiotherapy may be fast neutron radiotherapy. In the general case, fast neutron radiotherapy involves irradiating a certain volume in the subject's body with a neutron beam that causes cell death in the specified volume.

In photodynamic therapy, you can use, without limitation, any application, dose or method known to specialists in this field of technology as effective for treating or reducing the intensity of cancer. In general, photodynamic therapy involves the administration of a photosensitizing agent, which preferably accumulates in the neoplastic mass and makes the neoplasm sensitive to light, followed by irradiation of the tumor with light radiation of an appropriate wavelength. Under such irradiation, the photosensitizing agent catalyzes the formation of a cytotoxic agent, for example, singlet oxygen, which kills cancer

- 16 010437 cells. Methods of administration and devices used in photodynamic therapy are described in the work of Norreg, Laps! Opso1. 7: 12-219 (2000) and in US Pat. Nos. 6283957, 6071908, 6011563, 5855595, 5716595 and 5707401.

Without wishing to confine themselves to any particular theory of exposure, the authors of the present invention suggest that the active vitamin соединения compounds can increase the sensitivity of cancer cells to radiotherapy, and this increased sensitivity causes changes in the cellular mechanisms that regulate apoptosis and / or the cell cycle. The administration of an active vitamin соединения compound can not only enhance but also expand the use of radiotherapy to treat or reduce the intensity of a cancer that would otherwise not respond to current radiotherapy. In addition, increasing the sensitivity of cells to treatment can provide a lower dose of radiotherapy, which reduces the side effects associated with radiotherapy.

Radiotherapy can be used to destroy tumor cells before or after surgical treatment, before or after chemotherapy, and sometimes during chemotherapy. Radiotherapy can also be used to palliatively reduce cancer symptoms, for example, to reduce pain. The types of tumors that can be treated with radiotherapy include localized tumors that cannot be completely removed, as well as metastases and tumors whose complete removal would cause unacceptable functional or cosmetic defects or be associated with unacceptable surgical risks.

It should be understood that the specific radiation doses used to treat cancer, and the route of administration, depend on many factors. So, the dosage of radiation, which can be used in accordance with the methods of the present invention, is determined by the requirements of each specific situation. Dosage depends on factors such as tumor size, location of the tumor, age and gender of the patient, dosing frequency, the presence of other tumors, possible metastases, etc. Specialists in the field of radiotherapy can easily determine the dosage and method of introducing radiation for any particular tumor using literature, in particular Na11, E. 1 .. Vayuyuodu Gog Fe Vayuyuyodk1, 5 ^4b uyuyup, Irrtsoy \ U | Shat5 & Khuikn Rtsykyega, РЫ1айе1рЫа, РА, 2000; Sipyegkop, b. B. apy Terreg 1. E., ee§., C1shyu1 KaFayop Ops1odu, SygSyn Lgutdyope, Lopyop, Epdyapy, 2000; apy sgoksy, Ό. 8. Vyuoduya1 EGGes (5 oG VaFayop, 2 ^pyyyup , Asayetk Rge§8, 8ap Egapksso, SA, 1980. In some embodiments, the introduction of radiotherapeutic agents and irradiation can be performed with lower doses than those known to specialists in this field of technology, due to the additional or synergistic effect of the active vitamin соединения compound.

The dosage and frequency of administration of additional therapeutic agents are determined by the conditions of therapeutic efficacy. The dosage and frequency of administration of these agents usually vary according to factors specific to each patient, depending on the particular therapeutic agents being administered, the severity and type of pancreatic cancer, the route of administration, as well as the age, body weight, reaction and patient history. A person skilled in the art can choose the appropriate regimes, taking into account these factors and following, for example, the dosage described in the literature and recommended by RyukSzap'k Ee ^ to BeGegeps (56 ' ¹ ' e., 2002).

For animals with resectable cancer, the active vitamin соединение compound can be administered before and / or after surgery. Similarly, the administration of chemotherapeutic and radiotherapeutic agents or treatment procedures can be performed before and / or after surgery.

According to the present invention, treatment with an active vitamin соедин compound can be carried out at any time before, during or after the administration of chemotherapeutic or radiotherapeutic agents or treatment procedures. The exact period of treatment with an active vitamin соедин compound varies depending on the active vitamin соединения compound used, such as pancreatic cancer, the patient, and other related factors. The active vitamin Ό compound can be administered between 12 hours and 3 months before or after the administration of chemotherapeutic or radiotherapeutic agents or treatment procedures. The active vitamin Ό compound can be administered at least 1 day before or the next day after administration of chemotherapeutic or radiotherapeutic agents or treatment procedures, and within 3 months before or after administration of chemotherapeutic or radiotherapeutic agents or treatment procedures. In some embodiments, the methods of the invention comprise administering an active vitamin соединения compound once every 3, 4, 5, 6, 7, 8, 9, or 10 days for a period of 3 to 60 days before or after the administration of chemotherapeutic or radiotherapeutic agents or therapeutic procedures.

The administration of an active vitamin соединения compound can continue at the same time as the administration of chemotherapeutic or radiotherapeutic agents or treatment procedures. In addition, administration of the active vitamin Ό compound can continue after administration of chemotherapeutic or radiotherapeutic agents or treatment procedures.

In some embodiments, a method of administering an active vitamin соединения compound alone or in combination with chemotherapeutic or radiotherapeutic agents or therapeutic procedures can be repeated at least once. The method can be repeated as many times as required to achieve or maintain a therapeutic response, for example 1 to 10 times.

- 17 010437

At each repetition of the method, the combination of active vitamin Ώ and chemotherapeutic or radiotherapeutic agents or treatment procedures may be the same or different from those used in the previous repetition.

In addition, the period and route of administration of the active vitamin соединения compound may vary from one repetition to another.

In a preferred embodiment, the cancers are treated with combination chemotherapy as described in US Pat. Nos. 6,087,350 and 6,559,139. In this embodiment, for the treatment of hyperproliferative disease, the active vitamin соединения compounds are administered in combination with other pharmaceutical agents, in particular cytotoxic agents. Preferably, pretreatment of the hyperproliferative cells with active vitamin Ώ compounds, followed by exposure to cytotoxic agents, increases the effectiveness of cytotoxic agents. For example, a compound of active vitamin Ώ can be administered a day before the administration of a chemotherapeutic agent.

Animals that can be treated according to the present invention include all animals whose condition can be improved by taking the preparations in accordance with the present invention. Such animals include humans, domestic animals such as dogs and cats, and farm animals, in particular cows, pigs, sheep, goats, etc.

The following are illustrative, but not limiting examples of the method and compositions in accordance with the present invention. Other suitable modifications and adaptations of a variety of conditions and parameters commonly encountered in clinical therapy, and obvious to those skilled in the art, are within the scope and scope of the present invention.

EXAMPLES

Example 1. Relative chemical compatibility of calcitriol and selected components.

In this example, the relative chemical compatibility of calcitriol and the selected lipophilic, hydrophilic, and surfactant components was evaluated by measuring the isolated percentage (recovery) of unchanged calcitriol after storage at 40 ° C and at 60 ° C. The degree of calcitriol recovery was determined on the basis of high pressure liquid chromatography (HPLC, HPLC). The results are presented in table. one.

Table 1. The degree of extraction of calcitriol,%, prepared together with the selected components

Component Filler Time The degree of extraction,%, at 40 ° C The degree of extraction,%, at 60 ° C Lipophilic Corn oil 0 100.0 100.0 3 days 93.77 104.80 7 days 90.27 91.50 14 days 89.89 86.46 Soybean oil 0 100.0 100.0 3 days 96.44 94.56 7 days 98.46 98.57 14 days 96.66 93.15 Sunflower 0 100.0 100.0 oil 3 days 99.10 99.33 7 days 102.7 102.93 14 days 96.56 88.79 Vitamin E 0 100.0 100.0 3 days 128.56 160.79 7 days 0.00 0.00 14 days 102.29 65.02 M | d1yo1 812 0 100.0 100.0 3 days 98.23 97.01 7 days 99.31 96.78 14 days 99.17 99.48 M (d1uo1 812, 0.02% 0 100.0 100.0 BGA / BHT 3 days 98.41 97.83 7 days 97.43 98.17 14 days 98.72 102.15 Sar1ekh 200 0 100.0 100.0 3 days 99,20 97.28 7 days 100.14 97.68 14 days 108.83 101.15 kaga! as SS 0 100.0 100.0

- 18 010437

3 days 98.60 95.84 7 days 100.05 99.51 14 days 101.37 100.24 R idrophilic PEG 300 0 100.0 100.0 3 days 78.22 18.95 7 days 52.68 4.61 14 days 10.09 1.84 Propylene glycol 0 100.0 100.0 3 days 97.56 99.71 7 days 101.73 108.47 14 days 105.83 138.22 Surface-active substance Сеторпог Е1_Р 0 100.0 100.0 3 days 82.61 66.28 7 days 62.86 60.90 14 days 51.90 59.92 ShetorKog KN 40 25% in M | d1uo1 812 0 100.0 100.0 3 days 105.30 91.91 7 days 92.10 78.30 14 days 96.88 87.95 Razulobae 80 0 100.0 100.0 3 days 87.94 67.43 7 days 87.29 71.71 14 days 60.52 66.08 SESHS1KE 44/14 25% in M | d1uo1 812 0 100.0 100.0 3 days 98.70 107.68 7 days 101.55 83.06 14 days 100.96 98.11 Vitamin E TRC5 25% in M (d! Yo1 812 0 100.0 100.0 3 days 101.15 97.26 7 days 101.26 98.74 14 days 103.61 100.15 Lambin M 0 100.0 100.0 3 days 98.46 95.19 7 days 99.45 95.64 14 days 100.30 78.97 Rockhateg 188 25% in M | d! Wo! 812 0 100.0 100.0 3 days 116.42 76.47 7 days 126.39 116.67 14 days 126.79 83.30

From the obtained values of the degree of extraction it can be seen that the most compatible components are Μίβίγοί 812 (in the presence or absence of BHT and BHA), LaBaGas SS and Sar1ex 200 from the group of lipophilic components, propylene glycol from the group of hydrophilic components and vitamin E TPC8 and OEiC1KE 44/14 from groups of surfactants.

Example 2. The stability of liquid and semi-solid preparations of calcitriol.

I. Introduction

In this example, the stability of a calcitriol preparation containing an active vitamin соединения compound was measured in nine different types of preparations (four liquid preparations and five semi-solid preparations).

II. Calcitriol Preparation

A. Liquid preparations

Four liquid preparations of calcitriol (L-L4) were prepared containing the ingredients listed in table. 2. The finished composition contained 0.208 mg of calcitriol per 1 g of a liquid preparation.

- 19 010437

55.0 ~ 0 ~

Table 2. The composition of the liquid preparations of calcitriol

Ingredient Calcitriol

M "d1uo1 812

Сар1ех 200 1_аЬга (ас SS

1_1

0,0208

56.0

Oh! _2

0,0208

62.0

Oh sh

0,0208

ABOUT

Ι_4

Vitamin E TRS

1_a bn! N M

1,2-propylene glycol

OSH

BGA

22.0

14.0

9.0

0Ts5

0Ts5

Amounts are in grams.

B. Semi-solid preparations

Five semi-solid preparations of calcitriol (881-885) were prepared, which contained the ingredients listed in the table. 3. The finished composition contained 0.208 mg of calcitriol per 1 g of semi-solid preparation.

Table 3. The composition of semi-solid preparations of calcitriol

Ingredient 881 882 883 884 885 Calcitriol 0,0208 0,0208 0,0208 0,0208 0,0208 M | d1yo1 812 80.0 0 65.0 0 79.0 Sar1ekh 200 0 82.0 0 60.0 0 1_aLga1as SS 0 0 0 0 12.0 Vitamin Ε TP68 20,0 18.0 5,0 5,0 9.0 Basti m 0 0 0 0 0 CE1_iS1KE 44/14 0 0 30,0 35.0 0 OSH 0.05 0.05 0.05 0.05 0.05 BGA 0.05 0.05 0.05 0.05 0.05

Amounts are in grams.

C. Method for the preparation of liquid and semi-solid preparations of calcitriol

1. Preparation of delivery vehicles

The gram quantities of four liquid preparations of calcitriol (L1-L4) and five semi-solid preparations of calcitriol (881-885), are listed in table. 2 and 3, respectively, were prepared as follows.

The listed ingredients, with the exception of calcitriol, were mixed in a suitable glass jar and mixed until homogeneous. Vitamin E TRC8 and OEiSGKE 44/14 were heated and homogenized at 60 ° C before being weighed and added to the preparation.

2. Preparation of active drugs

Semi-solid delivery vehicles were heated and homogenized at a temperature of <60 ° C. Calcitriol in an amount of 12 + 1 mg was weighed and laid out in separate glass bottles with screw caps under dim light, one bottle was used for one preparation. (Calcitriol is sensitive to light, when working with calcitriol / its preparations, a dimmed / red light is necessary). The exact weight was recorded to 0.1 mg. As soon as calcitriol was placed in bottles, the lids were immediately closed. Then, using the following formula, the amount of each delivery vehicle required to bring the concentration to 0.208 mg / g was calculated:

C ^ / 0.208 = required mass of the delivery vehicle, where C _A = mass of calcitriol, mg, and

0.208 = final concentration of calcitriol, mg / g.

Finally, an appropriate amount of delivery agent was added to each of the respective bottles containing calcitriol. The preparations were heated (<60 ° C) with mixing to dissolve calcitriol.

ΙΙΙ. Calcitriol drug stability

Nine calcitriol formulations (L1-L4 and 881-885) were analyzed to determine the stability of the calcitriol component at three different temperatures. Each of the samples of nine preparations was kept at a temperature of 25, 40 and 60 ° C. Samples of all nine preparations aged at three

- 20 010437 temperatures, analyzed by HPLC after 1, 2 and 3 weeks. In addition, samples aged at 60 ° C were analyzed by HPLC after 9 weeks. The percentage of the initial calcitriol remaining at each measurement moment, determined for each sample, is shown in table. 4 (liquid preparations) and tab. 5 (semi-solid preparations).

* percentage of concentration at time zero

Table 5. The stability of semi-solid preparations

A drug Temperature Return * calcitriol (%) Week 1 Week 2 Week 3 Week 9 Semi-solid #one 25 ° C 98.5 98.9 99.8 But. 40 ° C 99.6 99.0 98.2 But. 60 ^p C 97.9 97.2 96.3 104.6 Semi-solid # 2 25 ° C 100.0 99.6 100,4 But. 40 ^p C 98.7 99.6 98.7 But. 60 ^p C 97.2 98.0 98.6 100.0 Semi-solid # 3 25 ° C 101,2 98.9 100,4 But. 40 ° C 100.0 98.7 98.8 But. 60 ° C 98.3 97.6 98.4 97.1 Semi-solid #four 25 ° C 100,2 99.0 99.6 But. 40 ° C 98.4 99,2 98.5 But. 60 ° C 96.8 97.7 97.7 103,4 Semi-solid #5 25 ° C 98.8 99,2 98.9 But. 40 ^and C 99.0 97.1 96.8 But. 60 ° C 96.8 96.7 96.0 97.7

* percentage of concentration at time zero

As can be seen from the table. 4 and 5, calcitriol remains relatively stable with a very low degree of degradation in all analyzed preparations (liquid and semi-solid).

Example 3. The study of the appearance of calcitriol preparations, their UV absorption and absorption in the visible range.

Calcitriol 1.1 and 883 were prepared prior to the start of this study and were stored at room temperature, protected from light. In the table. 6 shows the quantities of the ingredients from which these preparations were prepared.

- 21 010437

Table 6. Composition of calcitriol preparations used for absorption analysis

Before use, the preparations were heated to 55 ° C. Both preparations (liquid # 1 and semi-solid # 3) were well mixed in a vortex mixer, they looked like transparent liquids. Each calcitriol preparation (= 250 μl) was added to a 25 ml volumetric flask. The exact added masses were 249.8 mg for liquid # 1 and 252.6 mg for semi-solid # 3. Upon contact with glass, the semi-solid preparation # 3 hardened. Then deionized water was added to 25 ml mark and the solutions were mixed in a vortex mixer until smooth. At this point, the observed appearance and absorption of the resulting mixtures at 400 nm was determined using UV spectrophotometry and spectrophotometry in the visible range. Deionized water was used as a control solution and measurements were carried out at 400 nm. The absorption of each 5 samples was measured 10 times over 10 min. The results are summarized in table. 7. Both preparations obtained were white and opaque.

Table 7. Absorption values for preparations at 400 nm

Measurement Liquid # 1 Semi-solid # 3 one 2.4831 1,6253 2 2,5258 1.6290 3 2,5411 1,6309 four 2,5569 1,6328 5 2,5411 1,6328 6 2,5258 1,6347 7 2,5569 1,6328 8 2,5111 1.6366 9 2,5111 1.6366 10 2,5411 1,6328 Average 2,5294 1,6324 Κ8ϋ% (ΚβΙβΐϊνβ 31aps1ags1 Οβνϊθίϊοη, relative standard deviation) 0.91 0.21

Example 4. The diameter of the droplets of the emulsion formed by liquid and semi-solid delivery means (without calcitriol).

In this example, the average diameter of the droplets of the emulsion obtained after dilution of liquid (L1-L4) and semi-solid (881-885) delivery vehicles of emulsion preconcentrates (not containing calcitriol) by imitation of gastric juice (ILS) without enzymes was measured. The average droplet diameter was determined by light scattering. The appearance of preconcentrates and emulsions obtained was also noted visually. The results are shown in table. 8.

- 22 010437

Table 8. The diameter of the droplets of the emulsion formed by the delivery means of pre-concentrates of emulsions (without calcitriol)

A drug The appearance of the emulsion preconcentrate The ratio of preconcentrate: IZHS Average hydrodynamic diameter * The appearance of the emulsion AND Clear liquid 1: 1600 237 Opaque 1_2 Clear liquid 1: 1600 281 Opaque 13 Clear liquid 1: 1600 175 Opaque B4 Clear liquid 1: 1600 273 Opaque 881 Semi-solid 1: 2000 305 Opaque 882 Semi-solid 1: 2000 259 Opaque 883 Semi-solid 1: 2000 243 Opaque 834 Semi-solid 1: 2000 253 Opaque 835 Semi-solid 1: 2000 267 Opaque * average in nanometers

From the above results it is seen that the droplets (particles) formed by the preparations of emulsion preconcentrates are particles of submicron size, despite the opaque appearance of the emulsion.

Example 5. The diameter of the droplets of the emulsion formed by liquid and semi-solid preparations of calc citriol.

In this example, the average diameter of the emulsion droplets obtained after diluting liquid # 1 (S) and semi-solid # 3 (883) emulsion pre-concentrates of emulsions by simulating gastric juice (IZH) without enzymes was measured. The drugs used in this example contained calcitriol at a concentration of 0.2 mg / 1 g of the drug. The diameter of the droplets was determined by the scattering of light. The appearance of preconcentrates and emulsions obtained was also noted visually. The results are shown in table. 9.

Table 9. The diameter of the droplets of the emulsion formed by the drugs

A drug Preconcentrate ratio: IZHS Average hydraulic diameter * The appearance of the emulsion AND 1: 1600 257 Opaque 883 1: 2000 263 Opaque

* average in nanometers

Example 6. The dispersion of calcitriol from preconcentrates of emulsions ίη ugo.

In this example, the dispersion of the dispersion of calcitriol in various preparations in gelatin capsules was determined. A single capsule containing 250 mg of calcitriol preparation in a size-2 gelatin capsule (each capsule contained 0.2 mg of calcitriol / g of the preparation) was added to 200 ml of an enzyme-free gastric juice (IZH) simulated at 37 ° C with a paddle stirrer at 200 rpm Then, the samples were filtered through a 5 μm filter and the concentration of calcitriol was measured after 30, 60, 90, and 120 min using HPLC. The results are shown in table. 10.

- 23 010437

Table 10. The percentage of calcitriol obtained in the filtrate after dispersion in IHL and filtration through a 5 μm filter

A drug 30 minutes. 60 min 90 min 120 minutes Liquid # 1 106 103 86 68 Semi-solid # 3 109 99 73 53 The drug comparisons ^# 0 0 0 0

The reference preparation contained calcitriol at 0.2 mg / g dissolved in M1d1uo1 812 with 0.05% BHA and 0.05% BHT. This drug is similar to KOSATKOB, manufactured by Koske Lakoga1: ope8.

As can be seen from this example, the dispersion of calcitriol in the simulation of gastric juice from capsules containing both L1 and 883 is much higher than the dispersion from capsules containing the comparison drug (similar to KOSALTKOB produced by Koske Lakoga1og1e8).

Example 7. Plasma concentrations and pharmacokinetics of calcitriol in dogs.

In a pharmacokinetic study on dogs, dog plasma plasma calcitriol levels were compared after administration of 1.0 μg / kg of three different drugs: COSTAKOB, a liquid preparation (liquid # 1) and a semi-solid preparation (semi-solid # 3). Four dogs were dosed orally with 1.0 μg / kg COSTABOS semi-solid preparation or liquid preparation. If dogs were given more than one drug, each drug was administered at least seven days later.

For analysis of calcitriol levels, blood samples were taken before and after 0, 5, 1, 2, 4, 6, 8, 10, 12, 24, 36, and 48 hours after taking the drug. For the KOSATKOB group, blood samples were taken for clinical chemical analysis before taking the drug and after 24 and 48 hours after it, for liquid and semi-solid preparations, samples were taken before taking the drug and after 4, 24, 48, 72, 96 and 120 hours after it. The calcitriol content in the samples was determined using a radioimmunological study, and then subjected to pharmacokinetic analysis.

The time dependence of calcitriol concentration in plasma for three preparations is shown in FIG. one.

The final results of pharmacokinetic studies of calcitriol for one of the three drugs at a usual dose of 1.0 μg / kg are presented in table. 11-14.

Table 11: Summary of Calcitriol Specific for Dogs

Parameter KOSA1_TBYU1_ Semi-solid # 3 Liquid # 1 Average 8ϋ Average 8ϋ Average δϋ Stakh, PG / ML 717.4 51.5 2066.6 552.5 2164.4 253.9 Ttah, 430 3.0 (2-6) 2.0 (1-2) 1,5 (1-2) A11C (o- "), pg ' hour / ml 11,988.0 3804.7 12351.7 1624.9 14997.9 3531.7 Τι / 2 ^& , hour 25.1 11.1 4.8 1,2 1,2 3,5

(81apbagb Peu1aIop) is the standard deviation, ^a is expressed as the mean and range, ^b is expressed as the harmonic mean and pseudo 8ения based on the deviation calculated by the sampling method (а ш к к ап ап ап ап ап).

Table 12. Plasma concentration (pg / ml) and pharmacokinetic parameters of calcitriol in dogs after ~ one dose of COSABTKOB at a dose of 1 μg / kg

Parameter Time, hour Dog 101 Dog 102 Dog 103 Dog 104 Average 8ϋ 0,0 Will BO1 Will BO1 0 0 0.5 488.2 304.8 182.7 ΒΩί. 243.9 205,4 1,0 478.2 634.8 500,7 555.7 542.4 69.7 2.0 518.2 700.8 749.7 765.7 683.6 113.7

- 24 010437

4.0 494.2 658.8 750.7 745.7 662.4 119.8 6.0 652.2 566.8 496.7 532.7 559.9 68.0 8.0 381.2 366.8 418.7 381.7 387.1 22.2 10.0 313.2 212.8 165.7 158.7 212.6 71.2 12.0 190.2 186.8 189.7 171.7 184.6 8.7 24.0 78,2 78.8 69.7 97.7 81.1 11.8 36.0 63,2 83.8 80.7 67.7 73.9 10.0 48.0 66,2 47.8 45.7 52.7 53.1 9.2 Stach, pg / ml 652.2 700.8 750.7 765.7 717.4 51.5 Ttah, 430 6.0 2.0 4.0 2.0 3.0 (2-6) Ais _((M , PG hour / ml 17693.6 10094.5 9976.2 10187.5 11,988.0 3804.7 Τι / 2 ¹⁵ , hour 100,4 18.8 20,2 21.3 25.1 11.1

8Ό - standard deviation,

VOE - the value is below the limit of determination,

- expressed as the mean and range, ^b - expressed as the harmonic mean and pseudo 8Ό based on the deviation calculated by the sampling method (] no.

Bold font was used to calculate λ.

Table 13. Plasma concentration (pg / ml) and pharmacokinetic parameters of calcitriol in dogs after one dose of semi-solid preparation # 3 at a dose of 1 m kg / kg

Parameter Time hour Dog 101 Dog 102 Dog 103 Dog 104 Average 0,0 ALL ALL ALL ALL 0 0 0.5 198.1 11.0 all VOE 52.3 97.4 1,0 1208.1 2246.0 1128.7 503.4 1271.6 722.0 2.0 1255.1 2110.0 2269.7 2495,4 2032.6 541.9 4.0 902.1 1371.0 1095.7 1437.4 1201.6 248.5 6.0 603.1 1039.0 932.7 1112.4 921.8 224.9 8.0 815.1 441.0 593.7 848.4 674.6 192.4 10.0 253.1 489.0 285.7 305.4 333.3 106.0 12.0 213.1 295.0 184.7 170,4 215.8 55.7 24.0 50.1 37.0 40.7 29.4 39.3 8.6 36.0 14.1 ALL ALL 13.6 6.9 8.0 48.0 ALL all VOE ALL 0,0 0,0 Stach, pg / ml 1255.1 2246.0 2269.7 2495,4 2066.6 552.5 Ttah> Hour 2.0 1,0 2.0 2.0 2.0 (1-2) Ais ^), PG · h / ml 10333.8 14012.9 11813.8 13,246.4 12351.7 1624.9 Τι / ₂ ^ΰ , hour 6.2 3.8 4.1 5.9 4.8 1,2

8Ό - standard deviation,

VOE - the value is below the limit of determination, ^and - is expressed as the mean and range, ^b - is expressed as the harmonic mean and pseudo 8Ό based on the deviation calculated by the sampling method (] asksh e e wapse),

Bold font was used to calculate λ.

- 25 010437

Table 14. Plasma concentration (pg / ml) and pharmacokinetic parameters of calcitriol in dogs after a single dose, liquid preparation # 1 at a dose of 1 μg / kg

Parameter Time hour Dog 105 Dog 106 Dog 107 Dog 108 Average δϋ 0,0 ALL ALL Will ALL 0 0 0.5 ALL 57.6 523.0 350,0 232.7 246.9 1,0 1283.0 238.6 2266.0 2468.0 1563.9 1,024.0 2.0 2028.0 1895.6 2026.0 2373.0 2080.7 204.5 4.0 1090.0 892.6 1009,0 1771.0 1190.7 395.3 6.0 871.0 763.6 730.0 1063.0 856.9 150.0 8.0 301.0 579.6 374.0 562.0 454.2 138.1 10.0 421.0 520.6 464.0 517.0 480.7 47.4 12.0 348.0 290.6 170.0 373.0 295.4 90,4 24.0 42.0 165.6 62.0 202.0 117.9 78.0 36.0 49.0 111.6 ALL 79.0 59.9 47.4 48.0 35.0 15,5 ALL ALL 12.6 16.6 Stah! pg / ml 2028.0 1895.6 2266.0 2468.0 2164.4 253.9 Ttah ³ ! Hour 2.0 2.0 1,0 1,0 1,5 (1-2) AiSfo "), PG hour / ml 13474.4 14,296.3 12101.0 20117.7 14997.4 3531.7 Τι / ₂ ^ϋ , hour | 10.6 8.5 5,0 10.1 7.8 3,5

8Ώ - standard deviation,

In (') 1, - the value is below the limit of determination,

- expressed as the mean and range, ^b - expressed as the harmonic mean and pseudo 81) based on the deviation calculated by the sampling method (] asc1e wapapse), Bold font was used to calculate λ.

The results of the study show that there are the following differences and similarities in the pharmacokinetics of the drugs in accordance with the present invention and SCAN:

C _max for liquid and semi-solid preparations is approximately three times higher than C _max for SLIP;

With _max for liquid and semi-solid preparations, it is achieved faster (from 1 to 2 hours) than for the preparation SCAN (from 2 to 4 hours);

the total systemic effect (OuegaP 8u81et1s exrozige) (AIS _0-yes ) for all three drugs is comparable, although the systemic effect (8u81et1s exrozige) during the first 24-48 hours is greater for liquid and semi-solid preparations than for Kosatko.

The above results show that the highest values of C _max and AIS of calcitriol were obtained with the introduction of liquid preparation # 1, slightly lower values were obtained with the introduction of semi-solid preparation # 3. The smallest values of C _max and AIS are obtained for SLIDING. It turned out that liquid # 1 and semi-solid # 3 drugs are absorbed much faster and cause higher plasma concentrations during the first 12 hours, and also have a high excretion rate.

Example 8. The pharmacokinetics of semi-solid preparation # 3 after taking high doses.

In this study, the pharmacokinetics of the semi-solid preparation # 3 in dogs was studied after increasing the oral dose of the drug. Three males and three female hounds were administered orally at a time 0.5 μg / kg (to all six dogs), 0.1 μg / kg (1 male and 1 female), 5.0 μg / kg (2 males and 2 females) and 10.0 mcg / kg (all dogs) of the drug. After a dose of 10 mcg / kg, two dogs of each gender were euthanized. The remaining dogs of both sexes were continued to be examined and doses of 30 and 100.0 μg / kg were administered to them. After each dose, the dogs were kept for 6 days.

Blood samples (approximately 1 ml) were taken before and after 0, 2 (in all dogs, except those who were injected with 0.5 μg / kg), 4, 8, 24, 48 and 96 hours after taking the drug. The content of calcitriol in the samples was determined by radioimmunological studies, and then subjected to pharmacoka

- 26 010437 netic analysis. Plasma calcitriol concentrations are shown graphically for males and females in FIG. 2 A and 2B.

After the introduction of semi-solid preparation # 3, the maximum plasma concentrations were usually observed in a sample taken after 2 hours. It turned out that at doses greater than 0.1 μg / kg, the rate of decrease in the concentration of calcitriol in blood plasma was higher during the first 8 hours than in for 24 and 96 hours

At the lowest dose of 0.1 μg / kg, the concentration of calcitriol in the blood plasma fell below the detection limit after 24 hours. At a dose of 0.5 μg / kg and above, the measured concentrations of calcitriol were maintained even after sampling 96 hours after drug administration. There were no particular differences between the results obtained for males and for females.

The pharmacokinetic parameters for semi-solid preparation # 3 at doses ranging from 0.1 to 100.0 μg / kg are presented in table. fifteen.

Table 15: Pharmacokinetics of calcitriol after taking high doses of calcitriol (semi-solid preparation # 3)

Dose (mcg / kg) 0.1 0.5 5.0 Floor Male Female Male Female Male Female N one one 3 3 2 2 Stach (pg / ml) 566 473 1257 1431 17753 18346 Ttah (hour) 2.0 2.0 4.0 4.0 2.0 2.0 A11C (0-24) (pg hour / ml) 4311 2654 11431 15598 104027 107452 DiS (0-48) (pg hour / ml) 4311 2654 13584 19330 125408 126746 diS (o-oo) (pg · hour / ml) 4916 2718 15062 21644 200283 160681 T) / 2 (hour) 4.2 2.7 17.1 14.2 67.6 36.8 Dose (mcg / kg) 1C 1,0 AP ),about Jus ),about Floor Male Female Male Female Male Female N 3 3 one one one one Stakh (PG / ML) 23858 32336 53005 115896 238619 211631 Ttah (hour) 2.7 2.0 2.0 2.0 2.0 2.0 DiS (0-24) (pg hour / ml) 183981 203857 311841 567717 1165988 1089831 DiS (0-48) (pg hour / ml) 223977 240483 370713 641469 1381424 1256007 Ais ^ o ") (pg hour / ml) 388600 345936 531303 854841 1874997 1731873 Τι / 2 (hour) 77.7 56.0 56.3 58.2 45.3 53.7

These pharmacokinetic results indicate the following:

the systemic effect (zu81et1s exrozige) for calcitriol turned out to be quite linear in the entire range of doses tested, from 0.1 to 100.0 μg / kg. Neither saturation nor absorption was noticed;

The half-life of calcitriol, as it turned out, depends on the dose. Drugs having a half-life of more than 24 hours are less acceptable for use in a pulsed dosage regimen;

weekly dosage of semi-solid preparation # 3 when taking 5.0 μg / kg and above led to some accumulation in blood plasma. Such accumulation was not observed with sufficient constancy at lower doses of 0.1 and 0.5 μg / kg.

Example 9. The toxicity study of semi-solid preparation # 3 when administered orally to dogs for 28 days.

To determine the pharmacokinetics of calcitriol after a weekly oral administration of capsuliro

- 27 010437 of the given drug, a toxicological study of semi-solid preparation # 3 was carried out with repeated administration to dogs within 28 days. The capsules of the semi-solid preparation # 3 or the control drug were administered on the following days, counting from the start of the study: 0, 7, 14,21 and 28. Twelve dogs (6 males and 6 females) received the control drug - a delivery vehicle (group 1), eight dogs (4 males and 4 females) received 0.1 μg / kg of semi-solid preparation # 3 (group 2) and eight dogs (4 males and 4 females) received 1.0 μg / kg of semi-solid preparation # 3 (group 3). Twelve dogs (6 males and 6 females) received 30.0 μg / kg semi-solid preparation # 3 on day 0 of the study (group 4). Due to the acute clinical response observed after taking 30.0 μg / kg of the drug on day 0 of the study, the dose levels for this group were then reduced to 10 μg / kg (on days 7, 14, 21 and 28 for males) or 5 mcg / kg (on days 7, 14, 21 and 28 for females). Blood samples were taken from each dog before administering a dose of the drug and 1, 2, 4, 6, 8, 24, and 48 hours after it for 0 days of the study (first dose) and 21 days of the study (fourth weekly dose). All animals were euthanized on day 29 from the start of the study.

The pharmacokinetic results for calcitriol in blood plasma for groups 2-4 are presented in table. 16.

Table 16. Average toxicokinetic parameters for calcitriol after weekly intake of semi-solid preparation # 3 in dogs

Clothe Dose 0.1 mcg / kg (group 2) 1.0 mcg / kg (group 3) 30.0 mcg / kg (group 4) Gender (Νο dogs) Male (4) Female (4) Male (4) Female (4) Male (6) Female (6) Stakh, PG / ML 198.7 430.8 2380.0 3419.1 84909.1 57133.3 Ttah, 480 1,0 2.0 1,0 1,5 2.0 2.0 D11S (0-24), pg 1840.6 3093.4 17144.2 23259.7 496,044.6 323573,1  hour / ml di C (0-48), PG 2130.8 3093.4 19141.6 25794.5 644,064.2 365340.7  hour / ml 0 day Dose 0.1 mcg / kg 1.0 mcg / kg 10.0 mcg / kg 5.0 mcg / kg (group 2) (group 3) (group 4) (group 4) Gender (Νο Male (4) Female (4) Male (4) Female (4) Male (6) Female (6) siyaki) Dose 0.1 0.1 1,0 1,0 10D ¹⁵ 5D ⁵ Stah, 217.6 398.3 2272.1 2188.6 29061.8 8670.7 pg / ml Ttah, Chas 1,0 2.0 1,5 2.0 1,0 2.0 DiS (0-24), 1956.2 3283,0 19765,4 12,947.3 173597.2 46878.1 pg ’hour / ml diS (o-48), 2225.9 3640.7 24,606.9 15380.0 209732,1 54976.1 pg · hour / ml

but

- values

T ^x tah

- average values for this parameter. For all other parameters, the average values are also shown, ^b - the dose of semi-solid preparation # 3 was reduced starting from the 7th day of the study.

Data on the control group of dogs taking only the delivery vehicle (group 1) were not subjected to pharmacokinetic analysis.

In FIG. 3A and 3B show refined curves of the concentration of calcitriol in blood plasma versus time after oral administration of capsules of semi-solid preparation # 3 at 0 and on day 21 of the study by male (Fig. 3A) and female (Fig. 3B) beagle dogs. From all subsequent values of the concentration of calcitriol in the blood plasma, the values obtained at time 0 at 0 day of the study were subtracted to count from the endogenous (initial) level of concentration of calcitriol in the blood plasma.

The results of the study indicate the following:

after oral administration of capsules of semi-solid preparation # 3, the concentration of calcitriol in plasma

- 28 010437 IU of blood increase rather quickly, reaching a peak concentration in blood plasma within 2 hours; the rate of decrease in the concentration of calcitriol in the blood plasma is more important during the first 8 hours after administration than during a later period of time (24-48 hours), which may indicate a redistribution of calcitriol into the extravascular space, followed by a slow release of calcitriol back into vascular space. This observation is more evident at higher doses than when taking lower doses;

24 hours after taking 0.1 μg / kg of the drug, the concentration of calcitriol in blood plasma decreases almost to the initial level. However, at higher doses of calcitriol, residual calcitriol concentrations, depending on the dose, were still recorded at the last sampling (48 hours), although all values returned to their original level (before taking the drug) one week after taking the drug;

the values of C _max and A&C are practically proportional to the dose for all doses tested (0.130.0 μg / kg);

AiC values of _0-24 at a low dose at which no detectable adverse effect was observed (0.1 μg / kg) were in the range from 1840.6 to 3283.0 pg-h / ml;

AiCO _-24 values at an average dose, which was the maximum permissible dose (1.0 μg / kg), ranged from 12947.3 to 23259.7 pg-h / ml;

AiS values of _0-24 at doses at which weight loss and moderate signs of intoxication were observed ranged from 46878.1 pg-h / ml (5.0 μg / kg, females) to 173597.2 pg-h / ml ( 10.0 mcg / kg, males);

AiC values of _0-24 at doses at which mortality was observed (30.0 μg / kg) ranged from 323573.1 to 496044.6 pg-h / ml;

no persistent differences in pharmacokinetic parameters were observed depending on the sex of the animals.

In general, it turned out that animals tolerate calcitriol in the same way after the first dose and after repeated doses taken once a week, with some exceptions, such as higher values of C _max and A&C on day 0 compared to 21 days at a dose of 1.0 mcg / kg in females (dependence is not obvious to males).

Example 10. The study of acute toxicity of three different drugs.

To assess the toxicity of calcitriol preparations, during the behavior of the study described in example 7, some intravital parameters were recorded, including clinical chemistry parameters. Blood samples were analyzed for the content of calcium, phosphorus, blood urea nitrogen (Yobie igoda pIgodep, ΒϋΝ), glucose, albumin, bilirubin (total), aminotransferase aspartate (antigens, amyloidaseinase, A8T), alanine aminotransferase, alanine amoebase ), alkaline phosphatase (a1 kaipe pHozrbase, AR) and creatinine.

No clinical toxicity was observed in any dog with any of the three drugs.

Hypercalcemia was observed after taking 1.0 μg / kg of any of the three drugs. The average group and individual ranges of serum calcium levels for each of the three drugs are shown in table. 17.

Table 17. Average group serum calcium levels, mg / dl

- 29 010437

(10.44) 5ϋ 0.33 0.29 1.72 1,52 1.35 0.76 0.47 Range 9.7- 10.5 10.7- 10.8 12.2- 16,4 12.1- 15,5 10.8- 13.6 11.5- 13.1 12, Ο- Ι 3.0

^a - historical range;

^b - historical average;

* - average outside the historical range;

ND = not available (no serum sample was taken).

In addition to increasing the concentration of calcium, in all groups, an increase in the levels of LTP, A8T, ΒϋΝ and creatinine was observed.

In general, the results of this study show that not one dog was found to be associated with any of the drugs (SCAN, liquid or semi-solid) clinical signs;

hypercalcemia was observed after oral administration of 1.0 μg / kg of any of the three drugs;

the temporary course of hypercalcemia was similar when taking all three drugs within 48 hours, after 48 hours for the group that was injected with SCAN, blood was not sampled;

the severity of hypercalcemia was comparable when taking all three drugs, the highest serum calcium content (17.0 mg / dl) was observed in dogs treated with the liquid drug, 24 hours after administration;

observed that the average values of ABT, A8T, ΒϋΝ and creatinine were outside the historical range (previously obtained values) for all experimental groups at a single or several moments of analysis;

increased levels of ΒϋΝ and creatinine were greater for groups that were injected with a liquid or semi-solid preparation; in the absence of a competitive control group, the significance of this observation is not clear.

Example 11. The study of the maximum permissible high dose.

The study described in Example 8 also determined the acute toxicity and hypercalcemic effect of semi-solid preparation # 3 to evaluate the maximum allowable dose and to collect the data needed to select a dose for future studies.

Calcium levels increased depending on the dose at all dose values in male (Fig. 4A) and female (Fig. 4B) dogs. Data on the level of calcium in the blood serum of males at doses of 0.001 and 1.0 μg / kg were obtained in the study described in example 10, and were included in this example for completeness of the experiment.

In general, this study of semi-solid preparation # 3, administered orally in capsules to male and female beagle dogs at doses of 0.1, 0.5, 5.0, 10.0, 30.0 and 100.0 μg / kg, showed:

the most common laboratory abnormality was hypercalcemia, depending on the dose;

at doses of 5.0 μg / kg and higher, an increase in creatinine, urea nitrogen, cholesterol, erythrocytes, hemoglobin, hematocrit and neutrophils, as well as a decrease in lymphocyte levels, were observed;

after the administration of doses of 30.0 and 100.0 μg / kg, a significant decrease in the animal’s body weight and food intake was noted, after the intake of 100.0 μg / kg the dogs noticeably lost weight and their activity was reduced. Based on these results, the maximum allowable dose of semi-solid preparation # 3 for dogs was a dose of 5.0 μg / kg.

Example 12. Toxicological examination when taking repeated doses for 28 days.

In a study conducted as described in Example 9, the potential toxicity of semi-solid preparation # 3 for dogs by oral administration (capsules) was evaluated once every 7 days for 28 days. The study included the assessment of clinical signs, body weight, food intake, toxicokinetics, clinical pathology, including biochemical parameters, hematological parameters, coagulation (blood coagulation) and urinalysis, ophthalmic parameters, cardiac parameters, macroscopic autopsy, determination of organ mass and complete histopathology during all animals. The results are shown in table. eighteen.

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Table 18. Research plan for repeated doses of dogs for 28 days

Group The number of main (surviving) animals Input Materials Total Dose Level (mg / kg / dose) * Calcitriol Dose Level (mg / kg / dose) Males Females one 4 (2) 4 (2) Control medication 300 ** 0 2 four four Test Medication * one 0.1 3 four four Test Medication * 10 one four 4 (2) 4 (2) Test Medication * 300/100 (males) ** 300/50 (females) ** 30/10 (males) ** 30/5 (females) **

* Control medication (calcitriol semi-solid preparation # 3) - a preparation containing 0.1 mg of calcitriol per gram ** Doses were reduced to 10 μg / kg for males and 5 μg / kg for females at week 2, all surviving animals were euthanized on day 29.

Four animals from group 4 (1 male and 3 females) died or were killed before death during the first three days of the study. After lowering the dose level on day 7, the animals did not die, in groups 1, 2 or 3, the animals did not die at all.

In group 4, in animals that died, before death, the following most noticeable clinical deviations were noted: red vomiting, small / no feces, loose stools containing red material, red nasal discharge, shallow / rapid breathing, decreased activity and lateral lying position.

In animal groups 3 and 4, a decrease in body weight depending on the dose, a decrease in body weight gain and a decrease in the amount of food consumed were noted, the indicators in group 3 were -11-12% lower than the control indicators, in group 4 they were -17-24 ⁰ % below the benchmarks. In animal group 2, no apparent decrease in the weight or amount of food consumed was observed.

On day 29, in animals group 4, a tendency to increase in some parameters of red and white blood cells (air force apf ^ BC) was also observed, and toxicologically significant hematological abnormalities were not observed in groups of animals 3 and 4.

Dose-dependent hypercalcemia was observed in animal groups 3 and 4. Calcium levels reached elevated values 6 hours after taking the drug, reaching a maximum value 24 hours after taking the drug, and gradually decreased 48 and 96 hours after taking the drug. Other anomalies in clinical chemical parameters in animal groups 3 and 4 included elevated serum protein and cholesterol, increased renal function, and decreased specific gravity of electrolytes and urine. In group 2 animals, toxicologically significant anomalies of clinical chemical parameters or a significant increase in serum calcium were not observed.

On the 22/23 day of the study, no drug-induced changes in the eye tissues were observed; in this study, no drug-induced changes in the ECG and blood pressure were also observed.

The largest anomalies detected during macroscopic autopsy were observed for 4 groups of animals that were found dead or were euthanized, and they included damage to the digestive system and dependent organs, the dark red color of the omentums, the color of the mucous membranes of the esophagus and colon — from reddened to dark red, spotted and thickened gall bladder, blood clots in the heart, dark red and rash-covered areas of the lungs, the color of the pancreas from reddened to dark red, dark red thymus, thickened urine eva bladder and pale spleen. Macroscopic anomalies were less pronounced in the 3rd group of animals; in the 2nd group of animals, no noticeable macroscopic anomalies were noted.

The primary histopathological abnormality was dose-dependent interstitial nephritis: small to moderate in group 3 animals and moderate to severe in group 4 animals. Other microscopic findings in these groups of animals were: interstitial nephritis, from secondary to chronic, and included mineralization of various organ

- 31 010437 new / fabrics. No microscopic lesions were detected in group 2 animals.

The highest values of serum calcium concentrations usually appeared within the first 24 hours after taking the drug and again reached baseline levels before the next dosage. Sample data (males, on day 21) of serum calcium concentrations and plasma calcitriol concentrations are shown in FIG. 5A-5C. These data show that the maximum concentration of calcitriol in plasma is reached long before the maximum concentration of calcium in serum is reached.

In general, this study of semi-solid preparation # 3, administered orally every 7 days to male and female beagle dogs at doses of 0, 1.0 and 5.0 (females) or 10.0 (males) μg / kg, after an initial dose of 30 , 0 mcg / kg showed:

the dose level at which no adverse effect was observed was 0.1 μg / kg, the maximum allowable dose was 1.0 μg / kg, mortality was seen at 30 μg / kg;

in animal groups 3 and 4, dose-dependent damage to the digestive system and dependent organs, decreased weight gain and reduced food intake were observed;

in animal groups 3 and 4, dose-dependent interstitial nephritis was observed.

Example 13. Pharmacokinetic study in humans.

The pharmacokinetics of semi-solid preparation # 3, obtained in humans, was evaluated using a clinical trial. In this study, patients received a semi-solid preparation # 3 at doses of calcitriol up to 90 mg. Preliminary pharmacokinetic results are discussed below.

Blood samples were taken before and after 0.5, 1, 1.5, 2, 3, 4, 6, 8, 12, 24, 48 and 72 hours after the initial dose of semi-solid preparation # 3. Calcitriol content was determined using an available radioimmunoassay with limited dilution validation.

For each group, the curves of the dependence of the average plasma concentration on time were obtained (Fig. 6). The integrated (pop-soshrag1shep! A1) pharmacokinetic parameters were calculated for each subject, and then averaged (Table 19). From all subsequent values, the initial values were subtracted to count from the endogenous (initial) level of calcitriol concentration.

Table 19. Pharmacokinetic parameters of semi-solid preparation # 3 for the dose group

Dose, mcg Stach, pg / ml (± 50) Ttah, 430 (medium and range) AiS (0-24h), pg ’hour / ml (± 50) AiS (o_48h), pg hour / ml (± 50) Ais _(0. ~) Pg 'hour / ml (± 50) C / 2, hour * 5.0 (n = 3) 398.3 (12.9) 1.00 (1-1) 3665.7 ** 5627.3 *** (637.1) 5464.8 (892.8) 8.9 0,0 (n = 3) 898.8 (333.6) 1,50 (1,5-2) 6955.9 (2825.4) 9792.4 (2323.9) 11069.7 *** (1406.4) 16.3 *** 0,0 (n = 6) 2077.3 (533.3) 4.00 (1,5-4) 17480.6 (2,989.7) 20,999.4 (4762.5) 21,795.0 (5124.8) 7.3 0,0 (n = 4) 1918.4 (605.2) 1.3 (1-1.5) 17523.1 (1217.2) 20663.5 (1832.1) 24,997.6 (4612.5) 8.6 5.0 (n = 3) 1586.2 (328.6) 1,5 (1-4) 16,499.1 (2343.8) 21159.1 (3406.0) 22690.4 (9209,4) 10.8 0,0 (n = 3) 2858.7 (496.3) 1,5 (1-2) 23127.5 (5755.7) 28164.3 (8428.3) 29204.1 (9209,4) 8.8

* harmonic mean based on the deviation calculated by the sampling method (] askpye uapapse), ** n = 1, *** n = 2

Based on these data, it is shown that the pharmacokinetics of semi-solid preparation # 3 is linear and predictable. Saturation absorption was not observed.

Example 14. Safety data for the use of semi-solid preparation # 3.

The safety data for the use of semi-solid preparation # 3 for people was evaluated using a clinical trial. In this study, on May 8, 2002, 12 patients received semi-solid preparation # 3 as follows: 3 in group 1 (15 mcg), 3 in group 2 (30 mcg) and 6 in group 3 (60 mcg). Preliminary pharmacokinetic results in 9 patients are discussed below.

Not a single patient died. In 8 of 9 patients, 34 adverse effects were noted, 20 of

- 32 010437 adverse effects, apparently, were associated with the intake of semi-solid preparation # 3. One serious adverse effect, which the researcher believes is not related to taking the drug, was noted in group 3. This patient developed a passing class 1 fever on the first day, which increased the length of hospitalization. Adverse effects of classes 2 or 3, which, apparently, are associated with the study medication, are presented in table. twenty.

Table 20. Adverse effects of classes 2 or 3, which, apparently, are associated with the study medication

A patient Dose group Effect Sharpness Comments 002-1002 60 mcg Hyperglycemia Class 2 - Hypoproteinemia Class 2 - 002-1003 60 mcg Constipation Class 2 - Hyponatremia Grade 3 Sodium 127 meq / L on Day 4, Transitional, No Intervention

Preliminary results of the phase 1 trial of semi-solid preparation # 3 show: the maximum permissible dose of semi-solid preparation # 3 has not yet been determined in phase 1 of the trials, studies are ongoing on the status of additional patients in group 3 (60 mcg);

The semi-solid # 3 pharmacokinetics are linear and predictable for the first three dose groups.

Example 15. Additional formulations.

When semi-solid preparation # 3 was placed in hard gelatine capsules for oral administration, leakage of the composition from the capsules was observed. Therefore, we had to test new formulations containing various components of the lipophilic phase and surfactants, as well as different contents of each component, in order to find formulations that can solve this problem. The tested compositions are shown in table. 21.

Table 21. Additional tested formulations

Weight percent Structure but B from ό e ί 9 Ιί ϊ <3e1is1ge 44/14 fifty fifty fifty 40 40 thirty twenty 60 Vitamin E TRSV 10 twenty thirty twenty thirty thirty fifty 25 1УНд1уо1 812 fifty 40 thirty thirty thirty 40 fifty fifty fifteen

Weight percent Structure ΐ to I t η about R I am g Ce1is1ge 50/13 thirty fifty fifty fifty 40 40 thirty twenty 60 Vitamin E TRSV 5 10 twenty thirty twenty thirty thirty 25 M | d! Wo! 812 65 fifty 40 thirty thirty 40 40 fifty fifteen

- 33 010437

In addition, additional formulations containing several surfactants were tested without the lipophilic phase component. The compositions were a combination of vitamin E TPC8 with Oe1is1ge 44/14 or Oe1is1ge 50/13.

Identified compounds that do not give melt.

Example 16. Stable formulations for dosage forms.

Prepared compositions containing calcitriol to obtain the compositions shown in table. 22. Vitamin E TRC8 was heated to approximately 50 ° C and mixed in an appropriate ratio with ΜΙ6ΓΥΟΓ 812. BHA and BHT were added to each formulation to obtain a final concentration in the formulation of 0.35 wt.%.

Table 22. Compounds containing calcitriol

Composition number ΜΙΟΙ_ΥΟΙ_ (% by weight) TRSZ Vitamin E (% by weight) one one hundred 0 2 95 5 3 90 10 four fifty fifty

After preparation, formulations 2-4 were heated to approximately 50 ° C and mixed with calcitriol to obtain a concentration of 0.1 μg calcitriol / mg of the final formulation. Then, compositions containing calcitriol (~ 250 μl) were introduced into a 25 ml volumetric flask and deionized water was added to the 25 ml mark. After that, the solutions were mixed with shaking and the absorption coefficient was measured at 400 nm immediately after mixing (initial absorption) and 10 minutes after mixing. As shown in the table. 23, all three formulations after mixing with water formed an opalescent solution. Composition 4 formed a stable suspension in the absence of visible changes in the absorption coefficient at 400 nm after 10 minutes.

Table 23. The absorption coefficient of the compounds suspended in water

For further analysis of compounds containing calcitriol, a solubility study was conducted to determine the soluble calcitriol content in each formulation. Concentrations of calcitriol from 0.1 to 0.6 μg of calcitriol / mg of the composition were obtained by heating the compositions to 50 ° C followed by the addition of an appropriate mass of calcitriol. The compositions were then allowed to cool to room temperature and the presence of undissolved calcitriol was determined using an optical microscope with polarized light and without polarized light. For all formulations, calcitriol was soluble at a maximum experimental concentration of 0.6 μg / mg of formulation.

A dose of calcitriol of 45 μg is currently used in phase 2 clinical trials in humans. To obtain capsules with such a dosage, all formulations were prepared with a concentration of 0.2 μg calcitriol / mg composition and 0.35 wt.% BHA and BHT. No. 3 hard gelatin capsules were filled with 225 mg (45 μg calcitriol) bulk formulations. Then the stability analysis of the capsules was carried out at 5 ° C, 25 ° C / 60% KH (Ge1yu BitYNu, relative humidity), 30 ° C / 65% KH and 40 ° C / 75% KH. At the corresponding time points, the stability of the samples was determined by the content of whole calcitriol and dissolution of the capsules. The content of calcitriol in the capsules was determined by dissolving three open capsules in 5 ml of methanol and holding at 5 ° C until analysis. Then, the dissolved samples were analyzed by reverse phase HPLC. A RBetopech NuregzP ΒΏ8 C18 column was used at 30 ° С with an acetonitrile gradient from 55% acetonitrile in water to 95% acetonitrile at

- 34 010437 flow rates of 1.0 ml / min during elution. Peaks at 265 nm were determined by injecting 25 μl of sample in each cycle. The peak area of the sample and the reference was compared to calculate the calcitriol content, as shown in the table. 24. The dissolution test was carried out by placing one capsule in each of the six low-volume containers containing 50 ml of deionized water supplemented with 0.5% sodium dodecyl sulfate. Samples were taken 30, 60, and 90 min after mixing at 75 rpm and 37 ° C. The calcitriol content in the samples was determined by injection of 100 μl of the sample into a Be1a11 C18 column at a flow rate of 1 ml / min with a mobile phase containing 50:40:10 acetonitrile: water: tetrahydrofuran at 30 ° C (peak determination at 265 nm) . The average value of the results of dissolution after 90 min for six capsules is presented in table. 25.

Table 24. Chemical resistance of formulations containing calcitriol in hard gelatin capsules (total capsule filling mass 225 mg, 45 μg calcitriol)

Storage conditions Time (months) Analysis ³ (%) Composition 1 Composition 2 Composition 3 Composition 4 nd 0 100.1 98.8 99.1 100.3 5 ° C 1,0 99,4 98.9 98.9 104.3 25 ° C / 60% KH 0.5 99,4 97.7 97.8 102.3 1,0 97.1 95.8 97.8 100.3 3.0 95.2 93.6 96.8 97.9 30 ° C / 65% KH 0.5 98.7 97.7 96.8 100.7 1,0 95.8 96.3 97.3 100,4 3.0 94.2 93.6 95.5 93,4 40 ° C / 75% KH 0.5 96.4 96.7 98.2 97.1 1,0 96.1 98.6 98.5 99.3 3.0 92.3 92.4 93.0 96.4

a - the analysis results show% calcitriol relative to the expected value, based on 45 μg in the capsule. Values include predcalcitriol, which is the active isomer of calcitriol.

Table 25. Physical stability of formulations containing calcitriol in hard gelatin capsules (total capsule filling mass

a - capsules were dissolved as described above, and% calcitriol was calculated based on the standard and the expected calcitriol content of 45 mg per capsule. The active isomer, precalcitriol, is not included in the calculation of% dissolved calcitriol. The values presented refer to samples subjected to dissolution for 90 minutes.

The results of the determination of chemical resistance show that a decrease in the content of MULObO 812 with a concomitant increase in the content of vitamin E TPO8 provides an increase in the degree of extraction of whole calcitriol, as shown in table. 24. Composition 4 (50:50 MULO 812 / vitamin E TPO8) had the highest chemical resistance with a slight decrease

- 35 010437 degrees of extraction of whole calcitriol after 3 months of storage at 25 ° C / 60% KH, which allows its storage at room temperature.

The physical stability of the compositions was determined by the dissolution of the capsules after their storage in each mode of resistance testing. As in the test of chemical resistance, a decrease in the content of MUUEOE 812 and an increase in the content of vitamin E TPO8 improved the dissolution characteristics of the composition (Table 25). Composition 4 (50:50 MULO 812 / vitamin E TPO8) had optimal dissolution characteristics and suitable shelf life after storage at room temperature.

Example 17. Pharmacokinetics of the composition 4.

Experiments were conducted to compare the pharmacokinetic characteristics of composition 4 (# 4) from Example 16 and semi-solid composition # 3 (883). Formulations # 4 and 883 containing calcitriol were prepared and filled into capsules to obtain 4.5 μg of calcitriol per capsule. Separate capsules were administered orally to 20 male beagle dogs (dose was approximately 0.5 μg / kg body weight). Half of the dogs were given capsule # 4 on day 1 and capsule 883 on day 7. The other 10 dogs received capsule 883 on day 1 and capsule # 4 on day 7. Blood samples were taken 60, 40 and 20 minutes before each dose and after 0.5, 1, 1.5, 2, 3, 4, 6, 8, 10, 12, 24, 36, 48 and 96 hours after each dose . The results of the pharmacokinetic analysis of the level of calcitriol in blood samples are presented in table. 26.

Table 26. Comparison of the pharmacokinetics of calcitriol in formulations 883 and # 4.

Structure Stah (pg / ml) A11C (o- () (pg 'hour / ml) Ais _(0. ~) (Pg hour / ml) 833 1125 10061 11341 #four 1075 10269 11228

From the data presented, it can be seen that drugs # 4 and 883 have very similar pharmacokinetics and are therefore bioequivalent.

After becoming familiar with the full description of the invention, those skilled in the art should understand that such a study can be carried out in a wide range of equivalent conditions, preparations, and other parameters without limiting the scope of the invention and its implementation options. All patents, patent applications, and publications cited herein are hereby incorporated by reference in their entireties.

Claims (30)

CLAIM 1. A liquid or semi-solid pharmaceutical composition containing a compound of active vitamin Ώ, about 50% of the component of the lipophilic phase, about 50% of a surfactant, from about 0.05 to about 0.35% of butylated hydroxyanisole (BHA) and from about 0, 05 to about 0.35% butylated hydroxytoluene (BHT). 2. The pharmaceutical composition according to claim 1, characterized in that said active vitamin O compound is calcitriol. 3. The pharmaceutical composition according to claim 1, characterized in that said component of the lipophilic phase is MUUEOE 812. 4. The pharmaceutical composition according to claim 1, characterized in that said surfactant is vitamin E TPO8. 5. The pharmaceutical composition according to claim 1, containing a compound of active vitamin O, about 50% MUEEUOE 812 and about 50% vitamin E TPO8. 6. A unit dosage form containing a liquid or semi-solid pharmaceutical composition containing an active vitamin соединение compound, about 50% MUEUOE 812, about 50% vitamin E TPO8, about 0.05 to 0.35% BHA, and about 0.05 to approximately 0.35% OSH. 7. The standard dosage form according to claim 6, characterized in that said active vitamin O compound is calcitriol. 8. The unit dosage form according to claim 7, characterized in that the unit dosage form contains from about 10 μg to about 75 μg of calcitriol. 9. The unit dosage form of claim 8, wherein said unit dosage form contains about 45 μg of calcitriol. 10. The standard dosage form according to claim 6, characterized in that said component of the lipophilic phase is MUUEOE 812. 11. The standard dosage form according to claim 6, characterized in that said surfactant is vitamin E TPO8. 12. The unit dosage form according to claim 6, containing about 50% MUEUOE 812 and about 50% vitamin E TPO8. 13. The unit dosage form according to claim 6, characterized in that the unit dosage form contains about 45 μg of calcitriol, about 50% MUEEUOE 812, about 50% - 36 010437 Vitamin E TRC8. from about 0.05 to about 0.35% BHA; and from about 0.05 to about 0.35% BHT. 14. The unit dosage form according to claim 6, characterized in that said unit dosage form is a capsule. 15. The unit dosage form of claim 14, wherein said unit dosage form is a gelatin capsule. 16. The standard dosage form according to 14, characterized in that the total volume of ingredients in the specified capsule is 10-1000 μl. 17. A method of treating or preventing a hyperproliferative disease, comprising administering a pharmaceutical composition according to any one of claims 1-5 or a unit dosage form according to any one of claims 6-16 to a patient who needs it. 18. The method according to 17, characterized in that said hyperproliferative disease is cancer. 19. The method according to 17, characterized in that said hyperproliferative disease is psoriasis. 20. The method according to 17, characterized in that the administration of the drug is carried out in a pulsed dosage regimen, wherein said pulsed dosage regimen includes a patient receiving said composition no more than once every three days. 21. The method according to claim 20, characterized in that the reception is carried out no more often than once a week. 22. The method according to item 21, wherein the method is carried out no more often than once every three weeks. 23. The method according to any one of paragraphs.17-22, characterized in that it also includes the administration of one or more chemotherapeutic agents or radiotherapeutic agents / procedures. 24. The method according to claim 23, wherein said active vitamin соединение compound is taken at least 12 hours before the administration of said one or more chemotherapeutic agents or radiotherapeutic agents / procedures. 25. The method according to paragraph 24, wherein said active vitamin соединение compound is taken preliminarily from 1 day to about 3 months prior to administration of said one or more chemotherapeutic agents or radiotherapeutic agents / procedures. 26. The method according to item 23, wherein the specified compound of active vitamin Ό is taken simultaneously with the specified one or more chemotherapeutic agents or radiotherapeutic agents / procedures. 27. The method according to p. 26, characterized in that the intake of said active vitamin соединения compound is continued after the administration of said one or more chemotherapeutic agents or radiotherapeutic agents / procedures is completed. 28. The method according to item 23, wherein the intake of the indicated active vitamin соединения compound is carried out after the end of the administration of the indicated one or more chemotherapeutic agents or radiotherapeutic agents / procedures. 29. The method according to item 23, wherein said active vitamin соединение compound is taken 1 day before the administration of said one or more chemotherapeutic agents or radiotherapeutic agents / procedures. 30. The method according to item 23, wherein said active vitamin соединение compound and said one or more chemotherapeutic agents or radiotherapeutic agents / procedures are taken no more than once every three weeks.