Primary acid phosphate of prostaglandin d2 receptor antagonist

FIELD: medicine, pharmaceutics.

SUBSTANCE: present invention refers to a new acid dihydrogenphosphate of 2-(3-{6-[2-(2,4-dichlorophenyl)ethylamino]-2-methoxypyrimidine-4-yl}phenyl)-2-methylpropionic acid of formula optionally in a crystalline form exhibiting cAMP inhibitor properties. Also, the invention refers to a pharmaceutical composition.

EFFECT: compound can find application for treating the diseases associated with cell expression of prostaglandin D2 in such diseases, as allergic rhinitis, bronchial asthma, allergic conjunctivitis, etc.

3 cl, 12 dwg, 1 tbl, 1 ex

 

The scope of the invention

Serial production of pharmaceutical compositions poses many challenges for chemists and technologists. While many of the tasks associated with processing a large number of reagents and control chemical reactions in large volumes, one of the main tasks associated with the nature of the active end product. The product must not only be obtained with a high yield, to be stable and easily allocated, but also have properties that would be suitable for those types of pharmaceutical preparations in which it is to be used. The stability of the active ingredient of a pharmaceutical product should be ensured at every stage of the production process, including the synthesis, selection, storage, preparation drugs and dosage forms. At each of these stages can influence the state of the external environment (temperature and humidity).

For the preparation of pharmaceutical compositions should be used pharmaceutically active substances of very high purity, which also must have sufficient chemical stability to ensure reliable long-term storage of the drug in various conditions. This is absolutely necessary in order to prevent the formation of unwanted products RA the pada in pharmaceutical compositions because these products can be potentially toxic or lead to a decrease in activity of the composition.

The main task of the serial production of pharmaceutical compositions is the preservation of the polymorphic stability of the active substance in the course of its processing with the aim of ensuring the preservation of operating parameters and quality of the medicinal product. Depending on the characteristics of the stability of pharmaceutical compounds, the latter may be subjected to changes in the production process and/or storage that may impair the quality and technology of preparation of a medicinal product. Such changes may adversely affect the reproducibility of the production process and thus lead to the creation of preparative forms, which may not meet the stringent requirements for high quality requirements of the relevant authorities control technology preparation of medicines. Based on the above, we can conclude that, at least, the choice of pharmaceutical compounds with improved characteristics physical stability can provide a number of advantages of this form compared to other less stable forms.

The present invention pharmacologically active receptor antagonist of prostaglandin D2 relative to the Xia to monopotassium phosphate (salt), having the preferred physical properties. The connection is used as the DP antagonist for the treatment of a patient susceptible to pathological (disease) conditions mediated by PGD2, including, but not limited to, allergic disease (such as allergic rhinitis, allergic conjunctivitis, atopic dermatitis, bronchial asthma and food Allergy), systemic mastocytosis, disorders accompanied by systemic activation of mastocytes, anaphylactic shock, bronchoconstriction, bronchitis, urticaria, eczema, diseases accompanied by itch (such as atopic dermatitis and urticaria), diseases (such as cataract, retinal detachment, inflammation, infection and sleeping disorders)which arise as a secondary disease in the state, accompanied by itch (such as scratching and rubbing), inflammation, chronic obstructive pulmonary diseases, ischemic reperfusion injury, cerebral circulation disturbance, chronic rheumatoid arthritis, pleurisy, ulcerative colitis and similar diseases.

The level of technology

It is shown that local stimulation of allergen in patients with allergic rhinitis, bronchial asthma, allergic conjunctivitis and atopic dermatitis leads to rapid ro is the level of prostaglandin D2 (PGD2) in nasal and bronchial wash fluid, tears and skin abdominal fluid. PGD2 can have a variety of inflammatory action, for example to increase the permeability of blood vessels in the conjunctiva and the skin, to increase the resistance of the Airways of the nose, narrowing the respiratory tract and the infiltration of eosinophils into the conjunctiva and trachea.

PGD2 is the main ziklooksigenazny product of arachidonic acid produced by mastocytoma immunological stimulation [Lewis, R.A., Soter, N.A., Diamond P.T., Austen K.F., J.A. Oates, L.J. Roberts II, Prostaglandin D2 generation after activation of rat and human mast cells with anti-IgE,J. Immunol129, 1627-1631, 1982]. Activated mastocyte, one of the main sources of PGD2, play a key role in allergic reactions such diseases as asthma, allergic rhinitis, allergic conjunctivitis, allergic dermatitis and other diseases [C.E. Brightling, Bradding p, Pavord I.D., A.J. Wardlaw, New Insights into the role of the mast cell in asthma,Clin. Exp. Allergy33, 550-556, 2003].

Many of the effects of PGD2 mediated by its action at the receptor prostaglandin D (DP)-receptor-associated G-protein, expressed on the epithelium and smooth muscle.

In asthma respiratory epithelium has long been considered the main source of inflammatory cytokines and chemokines, which determine the development of the disease [Holgate, S., Lackie, P., Wilson, S., Roche, W., Davies D., Bronchial Epithelium as a Key Regulator of Airway Allergen Sensitization and Remodelling in Asthma,Am. J. Respir. Crit. Care Med.16 , 113-117, 2000]. In experimental models of asthma mice upon stimulation with antigen is a sharp activation of the DP receptor in the epithelium of the respiratory tract [Matsuoka, T., Hirata M., Tanaka H., Takahashi Y., Murata, T., Kabashima K., Sugimoto y, Kobayashi t, Ushikubi f, Aze y, Eguchi n, Urade Y., Yoshida N., Kimura K, Mizoguchi, A., Honda Y., Nagai H., Narumiya, S., prostaglandin D2 as a mediator of allergic asthma,Science287, 2013-2017, 2000]. In knockout mice with a null DP receptor is markedly reduced Hyper-reactivity of the Airways and chronic inflammation are two of the most important characteristic of asthma [Matsuoka, T., Hirata M., Tanaka H., Takahashi Y., Murata, T., Kabashima K., Sugimoto y, Kobayashi t, Ushikubi f, Aze y, Eguchi n, Urade Y., Yoshida N., Kimura K, Mizoguchi, A., Honda Y., Nagai H., Narumiya, S., Prostaglandin D2 as a mediator of allergic asthma,Science287, 2013-2017, 2000].

It is also believed that the DP receptor is involved in allergic rhinitis man, common allergic disease characterized by symptoms such as sneezing, itching, rhinorrhea, and nasal congestion. Local application of PGD2 in the nose causes a dose-dependent increase in nasal congestion [Doyle WJ, Boehm S, Skoner D.P., Physiologic responses to intranasal dose-response challenges with histamine, methacholine, bradykinin, and prostaglandin in adult volunteers with and without nasal allergy, J. Allergy Clin. Immunol.86(6 Pt 1), 924-35, 1990].

It was shown that the DP receptor antagonists reduce airway inflammation in experimental models of asthma Guinea pigs [Arimura, A., Yasui K., Kishino j, Asanuma F., Hasegawa h, Kakudo s, Ohtani M., Arita H., Prevention of allergic iflammation by a novel prostaglandin receptor antagonist, S-5751,J. Pharmacol. Exp. Ther.298(2), 411-9, 2001]. Thus, PGD2, apparently, acts on the DP receptor and plays an important role in the manifestation of the main features of allergic asthma.

It has been shown that antagonists of DP can effectively reduce the symptoms of allergic rhinitis in many species, in particular, it was shown that they can suppress induced antigens stuffy nose, the most obvious symptom of allergic rhinitis [Jones, T.R., Savoie, C., Robichaud, A., Sturino, C., Scheigetz, J., Lachance, N., Roy, B., Boyd, M., Abraham, W., Studies with a DP receptor antagonist in sheep and guinea pig models of allergic rhinitis,Am. J. Resp. Crit. Care Med.167, A218, 2003; Arimura, A., Yasui K., Kishino j, Asanuma F., Hasegawa h, Kakudo s, Ohtani M., Arita H., Prevention of allergic inflammation by a novel prostaglandin receptor antagonist, S-5751.J. Pharmacol. Exp. Ther.298(2), 411-9, 2001].

The DP antagonists are also effective in experimental models of allergic conjunctivitis and allergic dermatitis [Arimura, A., Yasui K., Kishino j, Asanuma F., Hasegawa h, Kakudo s, Ohtani M., Arita H., Prevention of allergic inflammation by a novel prostaglandin receptor antagonist, S-5751,J Pharmacol. Exp. Ther.298(2), 411-9, 2001; and Torisu K., Kobayashi K., Iwahashi M., Nakai Y., Onoda, T., Nagase, T., Sugimoto I., Okada Y., Matsumoto R., Nanbu F., Ohuchida s, Nakai H., Toda M., Discovery of a new class of potent, selective, and orally active prostaglandin D2receptor antagonists,Bioorg. & Med. Chem.12, 5361-5378, 2004].

In the patent application WO 2006044732 (hereinafter "the application '732"), which is in the form of references included in the present application, discloses pyrimidines, which have useful pharmaceutical SV is Ista, including, in particular, the ability to bind to DP receptor and regulate it. In the application '732 disclosed pyrimidines of the formula (I)

receipt containing these compounds, pharmaceutical compositions and their pharmaceutical use in the treatment of pathological conditions that can be controlled by inhibition of the receptor for prostaglandin D2. In addition, the application '732 described 2-(3-{6-[2-(2,4-dichlorophenyl)ethylamino]-2-methoxypyridine-4-yl}phenyl)-2-methylpropionate acid (hereinafter “free form”). In the application '732 is also given a General description of a wide variety of salts formed when attaching acids and bases, and to compounds that are the subject of the present invention, the above demonstration of the preparation of hydrochloric and sodium salts, more particularly sodium salts of hydrochloric and 2-(3-{6-[2-(2,4-dichlorophenyl)ethylamino]-2-methoxypyridine-4-yl}phenyl)-2-methylpropionic acid. However, in the application '732 was not disclosed information about salt, monopotassium phosphate 2-(3-{6-[2-(2,4-dichlorophenyl)ethylamino]-2-methoxypyridine-4-yl}phenyl)-2-methylpropionic acid.

Brief description of the invention

The present invention relates to monopotassium phosphate salt of 2-(3-{6-[2-(2,4-dichlorophenyl)ethylamino]-2-methoxypyridine-4-yl}phenyl)-2-methylpr pianoboy acid of formula (III) (hereinafter "monopotassium phosphate salt")

Another aspect of the present invention is a pharmaceutical composition comprising a pharmaceutically effective amount of monopotassium phosphate salt.

Another aspect of the present invention is a method of treating a patient suffering from disorder mediated by PGD2, including, in particular, allergic disease (such as allergic rhinitis, allergic conjunctivitis, atopic dermatitis, bronchial asthma and food Allergy), systemic mastocytosis, disorders accompanied by systemic activation of mastocytes, anaphylactic shock, bronchoconstriction, bronchitis, urticaria, eczema, diseases accompanied by itch (such as atopic dermatitis and urticaria), diseases (such as cataract, retinal detachment, inflammation, infection and sleeping disorders)which arise as a secondary diseases in the state, accompanied by itch (such as scratching and rubbing), inflammation, chronic obstructive pulmonary diseases, ischemic reperfusion injury, cerebral circulation disorders, chronic rheumatoid arthritis, pleurisy, ulcerative colitis and similar diseases, through the introduction of patient pharmaceutically effective what about the amount of the primary acid salt of 2-(3-{6-[2-(2,4-dichlorophenyl)ethylamino]-2-methoxypyridine-4-yl}phenyl)-2-methylpropionic acid.

A more detailed discussion of the present invention and the corresponding explanatory drawings below.

Brief description of drawings

Figure 1. Powder x-ray (CR) monopotassium phosphate (salt).

Figure 2. Table of values of interplanar distances d and relative intensities of peaks in the powder x-ray presented in figure 1.

Figure 3. thermogram of monopotassium phosphate, obtained by differential scanning calorimetry and thermogravimetric analysis (DSC-TGA). Data TGA show an overall loss of mass, which is about 0.4%when heated from room temperature to 175°C. The DSC contains information about the melting of the crystalline phase. The decomposition of the original substance begins at the melting temperature.

Figure 4. DSC-thermogram of monopotassium phosphate. From thermogram shows that the melting of the substance begins to occur at a temperature of 219°C.

Figure 5. Sorption isotherm monopotassium phosphate, obtained by using the analyzer's dynamic sorption of water vapor (DSWP), and the corresponding data table for the profile of water sorption. The measurement results show the increase in weight of the sample during sorption (approximately 0.9%) at a relative humidity of 94% and negligible hysteresis, which can be associated with the ABA connection with the surface water absorption.

6. Powder x-ray monopotassium phosphate before and after the procedure dynamic sorption of water vapor. The sample is prepared and recording x-rays begin within 2 minutes after removing the sample from the analyzer dynamic absorption of water vapor, where he for ~12 hours was at zero relative humidity. The obtained data show a slight change in intensities of the peaks and the interplanar distance d in the experiment DSWP, which means the absence of any significant changes in the crystal structure.

Fig.7.Micrograph monopotassium phosphate. The surface consists mainly of particles of elongated shape and in the form of needles with a length of approximately 30 microns.

Fig.Micrograph monopotassium phosphate before and after grinding and quantitative distribution of particle sizes, showing that during the grinding particles physico-chemical properties of the substance do not change. The photographs after grinding substances leads to the conclusion that monopotassium phosphate is fine. Measurements made approximately thirty different positions at 100 times magnification, shows that the particle size of the crystalline phase does not exceed 10 microns. It is established that the distribution of RA which measures particles of micronized monopotassium phosphate onemodule. The average particle size [x(50)] is 2.0 microns, 90% of all particles have a size of 4.7 microns or less. The micronisation process thus allows to reduce the average particle size (5.8 microns to grinding) and the size of 90% of the particles (before grinding was approximately 16 microns). Powder x-ray, i.e. the (intensity of the peaks, their position (interplanar distance (d) and width) before and after micronisation monopotassium phosphate salt is not changed.

Fig.9. Isotherm sorption of water vapor monopotassium phosphate after micronisation demonstrates the lack of amorphization.

Figure 10. Infrared Fourier spectrum of monopotassium phosphate and the corresponding table of the FTIR peaks.

11.The combination of powder x-ray patterns for the two samples of sodium salt, prepared in different ways. Sample 1 was prepared by recrystallization from a mixture of ethanol/ethyl acetate, sample 2 - by recrystallization from methanol/ethyl acetate.

Fig. The combination of powder x-ray patterns for the two samples hydrochloric salt prepared by recrystallization from acetone.

Detailed description of the invention

Definitions and abbreviations

Used above and throughout the text description of the invention, the following abbreviations unless otherwise specified, have the following meanings:

DMSO, dimethylsulfoxid the

cAMP cyclic monophosphate

IBMX 3-isobutyl-1-methylxanthines

SPA SPA assay (scintillation analysis)

ATTC American type culture collection

IPU minimum support environment

FBS fetal calf serum

pulse/min number of pulses per minute

Used above and throughout the description of the invention, the following terms, unless otherwise indicated, have the following meanings.

"Treatment" or "therapy" means the prevention, partial alleviation or cure of disease. Connection and preparation of the present invention are useful in the treatment of disorders mediated by PGD2, including, in particular, allergic diseases (e.g. allergic rhinitis, allergic conjunctivitis, atopic dermatitis, bronchial asthma and food Allergy), systemic mastocytosis, disorders accompanied by systemic activation of mastocytes, anaphylactic shock, bronchoconstriction, bronchitis, urticaria, eczema, diseases accompanied by itch (such as atopic dermatitis and urticaria), diseases (such as cataract, retinal detachment, inflammation, infection and sleeping disorders)which arise as a secondary disease resulting state, accompanied by itch (such as scratching and rubbing), inflammation, chronic obstructive Zab is lung diseases, ischemic reperfusion damage, disorders of cerebral circulation, chronic rheumatoid arthritis, pleurisy, ulcerative colitis and similar diseases, through the introduction of the patient pharmaceutically effective amount of the compounds of formula (III).

"Patient" means humans and other mammals.

"Pharmaceutically effective amount" means an amount of a compound, composition, medicament or other active ingredient that is effective to obtain the desired therapeutic effect.

Specific embodiments of the present invention

One specific embodiment of the present invention is a compound of formula (III) in crystalline form.

The compound forming the subject of the present invention is an antagonist of the receptor for prostaglandin D2 and can be used as an active pharmaceutical substance. Accordingly, it is included in the pharmaceutical compositions and used to treat patients suffering from certain medical disorders.

The compound forming the subject of the present invention is an antagonist of the receptor for prostaglandin D2 according to tests described in the literature and are described in the following section is about the pharmacological studies, the results are considered to correlate to pharmacological activity in humans and other mammals. Thus, in another embodiment, the present invention presents a connection, which is the subject of the present invention and compositions containing them, which can be used in the treatment of patients suffering from disease or susceptible to diseases, which can facilitate the introduction of PGD2 antagonist. For example, the compound forming the subject of the present invention may be used to treat a variety of disorders mediated by PGD2, including but not limited to, allergic disease (such as allergic rhinitis, allergic conjunctivitis, atopic dermatitis, bronchial asthma and food Allergy), systemic mastocytosis, disorders accompanied by systemic activation of mastocytes, anaphylactic shock, bronchoconstriction, bronchitis, urticaria, eczema, diseases accompanied by itch (such as atopic dermatitis and urticaria), diseases (such as cataract, retinal detachment, inflammation, infection and sleeping disorders)which arise as a secondary diseases in the state, accompanied by itch (such as scratching and rubbing), inflammation, chronic obstructive easy is x, ischemic reperfusion injury, disorders of cerebral circulation, chronic rheumatoid arthritis, pleurisy, ulcerative colitis and similar diseases.

One private implementation of therapeutic methods that are the subject of the present invention is the treatment of allergic rhinitis.

Another private implementation of therapeutic methods that are the subject of the present invention is the treatment of bronchial asthma.

Another private implementation of therapeutic methods that are the subject of the present invention is the treatment of COPD.

Another private implementation of therapeutic methods that are the subject of the present invention is the treatment of allergic conjunctivitis.

Another private implementation of therapeutic methods that are the subject of the present invention is the treatment of atopic dermatitis.

Included in the description links to treatment include both preventive therapy and treatment of established disease.

In addition, the connection forming the subject of the present invention may be used for treatment in combination with one or more of the following medications:

(i) antihistamines, such as Fexofenadine, desloratadine, loratadine, cetirizine, to ensure lergicescoe rhinitis;

(ii) leukotriene antagonists such as montelukast and shapirolast, for the treatment of allergic rhinitis, COPD, allergic dermatitis, allergic conjunctivitis, etc. - accurate information in the claims WO 01/78697 A2;

(iii) beta-agonists such as albuterol, albuterol and terbutaline to treat asthma, COPD, allergic dermatitis, allergic conjunctivitis, etc.;

(iv) antihistamines, such as Fexofenadine, loratadine, cetirizine, for the treatment of asthma, COPD, allergic dermatitis, allergic conjunctivitis, etc.;

(v) inhibitors of PDE4 (phosphodiesterase 4), such as roflumilast and cilomilast, for the treatment of asthma, COPD, allergic dermatitis, allergic conjunctivitis, etc.; or

(vi) antagonists TP (thromboxane A2 receptor or antagonists of CrTh2 (molecules homologous to receptor chemoattractant expressed on Th2 cells), such as ramatroban (BAY u3405), for the treatment of COPD, allergic dermatitis, allergic conjunctivitis, etc.

The present invention relates also to pharmaceutical compositions comprising a pharmaceutically effective amount of the compound which is the subject of the present invention, in a mixture with a pharmaceutically acceptable carrier.

In practice, the compounds forming the subject of the present invention, can enter the sterile in the form of pharmaceutically acceptable dosage forms for humans and other mammals by local or systemic application, including oral, inhalation, rectal, nasal, buccal, sublingual, vaginal, intestinal, parenteral (including subcutaneous, intramuscular, intravenous, intradermal, intrathecal and epidural), intracisternal and intraperitoneal. You should take into account that the specific route of administration may vary, for example, depending on the physiological state of the patient.

"Pharmaceutically acceptable dosage forms" refers to dosage forms of the compounds constituting the subject matter of this invention, which include, for example, tablets, pills, powders, elixirs, syrups, liquid formulations, including suspensions, sprays, inhalants, pills, pellets, emulsions, solutions, granules, capsules and suppositories, liquid formulations for injection, including liposomal drugs. A General description of the methods and compositions can be found in the latest edition Remington's Pharmaceutical Sciences, Mack Publishing Co., Easton, PA.

A special aspect of the present invention provides for the introduction of compounds that constitute the subject of the present invention, in the form of pharmaceutical compositions.

Pharmaceutically acceptable carriers include at least one component selected from the group comprising pharmaceutically acceptable carriers, diluents, shell, adjuvants, e is Scipioni or media, such as preservatives, fillers, disintegrating agents, wetting agents, emulsifiers, stabilizers of emulsions, suspendresume substances, isotonic agents, sweeteners, flavorings, fragrances, dyes, microbicides, antifungal agents, other therapeutic agents, lubricants, chemicals, slowing down or accelerating the absorption and spray substances, depending on the characteristics of the route of administration and dosage forms.

Examples suspendida substances are ethoxylated isostearyl alcohols, polyoxyethylenated and esters sorbitan, microcrystalline cellulose, Metagalaxy aluminum, bentonite, agar-agar and tragakant or mixtures of these substances.

Examples of bactericidal and antifungal substances that prevent the action of microorganisms, are parabens, chlorobutanol, phenol, sorbic acid and similar substances.

Examples of isotonic substances are sugars, sodium chloride and similar substances.

Examples of substances that slow down and prolong the absorption are aluminum monostearate and gelatin.

Examples of substances accelerate and stimulate absorption, are dimethyl sulfoxide and its analogs.

Examples of diluents, solvents, carriers, solubilizing agents, emulsifiers and the article is of bilization emulsions are water, chloroform, sucrose, ethanol, isopropyl alcohol, ethylcarbonate, ethyl acetate, benzyl alcohol, tetrahydrofurfuryl alcohol, benzyl benzoate, polyols, propylene glycol, 1,3-butyleneglycol, glycerin, glycols, dimethylformamide, Tween® 60, Span® 60, cetosteatil alcohol, ministerului alcohol, glycerylmonostearate and sodium lauryl sulfate, esters sorbitan and fatty acids, vegetable oils (such as cottonseed oil, peanut oil, olive oil, castor oil and sesame oil) and injectable organic esters, such as etiloleat and the like, or suitable mixtures of these compounds.

Examples of formative fillers are lactose, milk sugar, sodium citrate, calcium carbonate and dicalcium phosphate.

Examples of disintegrating agents include starch, alginic acid and certain complex silicates.

Examples of lubricants are magnesium stearate, sodium lauryl sulphate, talc, and polyethylene glycols of high molecular weight.

The choice of pharmaceutically acceptable carrier, in General, determined in accordance with the chemical properties of the active compound, such as solubility, mode of application and instructions that must be observed in pharmaceutical practice.

Pharmaceutical compositions forming the subject of this izopet the tion, suitable for oral administration may be a separate unit, such as solid dosage forms such as capsules, pills or tablets, each of which contains a certain amount of the active ingredient, or such as powders or granules, or liquid dosage forms such as solutions or suspensions in aqueous or non-aqueous liquid medium, or a liquid emulsion of oil in water or water in oil. The active ingredient can also be in the form of bolus, electuary or paste.

"Solid dosage form" means a dosage form of the compounds constituting the subject matter of this invention, in the form of solids, for example capsules, tablets, pills, powders, pills, or granules. In such dosage forms, the compound forming the subject of this invention, is added in at least one traditionally used excipient (or carrier)such as sodium citrate or dicalcium phosphate or (a) fillers or environment, such as, for example, starch, lactose, sucrose, glucose, mannitol and silicic acid, (b) binders, for example carboxymethylcellulose, alginates, gelatin, polyvinylpyrrolidone, sucrose and the Arabian gum, (c) moisturizing agents, such as, for example, glycerol, (d) disintegrating agents, such as, for example, agar-agar, calcium carbonate, potato sludge is manioc starch, alginic acid, certain complex silicates and sodium carbonate, (e) solutions-retardants, such as, for example, paraffin, (f) absorption accelerators, such as Quaternary ammonium compounds, (g) wetting agents such as, for example, cetyl alcohol and glycerylmonostearate, (h) adsorbents, such as kaolin or bentonite, and (i) moving substances such as, for example, talc, calcium stearate, magnesium stearate, solid polyethylene glycols, sodium lauryl sulfate, (j) fogging components, (k) a buffering agent, otsrochennoe releasing compound constituting the object of the present invention, in a certain part of the intestinal tract.

A tablet may be prepared by compressing or molding and may have one or more auxiliary components. Molded tablets can be obtained by compressing in a suitable machine the active ingredient in granular form, such as powder or granules, which may be mixed with a binder, a sliding agent, inert diluent, preservative, surface active or dispersing agent. Can be used such inert fillers as lactose, sodium citrate, calcium carbonate, dicalcium phosphate and leavening agents such as starch, alginic acid and certain complex silicates, mixed with gliding is their substances, such as magnesium stearate, sodium lauryl sulfate and talc. A mixture of powdered compounds moistened with an inert liquid diluent, can be molded in a suitable machine to obtain a molded tablets. Tablets may be uncoated or notches, or may have a composition that provide slow or controlled allocation of the contained active ingredient.

Solid compositions may also be used as fillers in soft and hard gelatin capsules using such inert excipients as lactose or milk sugar, as well as polyethylene glycols of high molecular weight and similar substances.

If necessary, as well as for more efficient allocation of the connection can be microencapsulation systems slow or directed release (or attached to such systems), such as a biocompatible, biodegradable polymeric matrix (e.g., copolymer of d,l-lactide with glycolide), liposomes and microspheres for subcutaneous and intramuscular injection method, called subcutaneous or intramuscular injection, slow absorption, providing a continuous slow release of the compound or compounds in a period of 2 weeks or longer. Connections can be sterilized, for example, by filtration through zadereev the store bacteria filter or adding sterilizing agents in the form of sterile solid compositions, which can be dissolved in sterile water or other sterile injectable medium immediately before use.

Liquid dosage form" means a form of active compound that is administered to the patient in liquid form, for example in the form of a pharmaceutically acceptable emulsions, solutions, suspensions, syrups or elixirs. In addition to the active compounds, the liquid dosage forms may contain commonly used in this field inert diluents, such as solvents, solubilizing agents and emulsifiers.

Aqueous suspensions can contain emulsifying agents or substances promoting the formation of a suspension.

Pharmaceutical compositions suitable for topical application, is a composition in the form, allowing for local administration to the patient. The compositions can be in the form of ointments, topical application, balms, powders, sprays and inhalants, gels (water or alcohol based), creams, usually used in this field, or to be included in the matrix base for use as a patch that provides a controlled release of compound through the skin barrier. In the form of an ointment, the active ingredients can be used with paraffin or water-soluble bases. Alternatively, the active ingredients can be in the form of a cream with an oil-water basis. ostavi, intended for local use through eyes that are eye drops, in which the active ingredient is dissolved or suspended in a suitable medium, usually an aqueous solvent. Formulations intended for local use through the oral mucosa include medicinal candy, having in its composition the active ingredient in the flavor additive, normally sucrose and gum or tragakant; lozenges, having in its composition the active ingredient in an inert basis such as gelatin and glycerin, or sucrose and gum; and compositions for rinsing of the mouth, having in its composition the active ingredient in a suitable liquid carrier.

The oil phase of the emulsion pharmaceutical compositions can be obtained in the usual way from the well-known ingredients. This phase can be composed of only the emulsifier (also known emulsifying substance), however, it is desirable that it also included at least one emulsifier containing fat or oil, or emulsifier-containing fat and oil. In a particular implementation of the invention, the formulation includes a hydrophilic emulsifier, together with a lipophilic emulsifier, which acts as a stabilizer. The emulsifier(s) with stabilizer(s) or without form emulgirujushchie wax together with the oil or fat to form emulgirujushchie basis is the ointment, which is the oily dispersed phase of the cream formulations.

If necessary, the aqueous phase of the cream base may include, for example, not less than 30 wt.% a polyhydric alcohol, i.e. an alcohol having two or more hydroxyl groups such as propylene glycol, butane-1,3-diol, mannitol, sorbitol, glycerol or polyethylene glycol (including PEG 400) and mixtures thereof. Compositions for topical application may, if necessary, to have a connection, stimulating the absorption or penetration of the active ingredient through the skin or other affected area.

The choice of suitable oils or fats for use in the composition depends on the properties that you want to retrieve. So, in particular, the cream should be non-greasy, does not leave stains and washable product with suitable consistency, preventing leakage from tubes or other containers. Can be used with linear and branched one - and dibasic alkalemia esters, such as diisopropylamide, decillia, isopropyl, butilstearat, 2-ethylhexylamine or mixture of esters with branched chain, known as Crodamol CAP. They can be used separately or in mixtures, depending on the desired properties. Alternatively can be used lipids with high melting point, such as white soft paraffin and/or liquid paraffin, and the other is their mineral oil.

Pharmaceutical compositions for rectal or vaginal application of the mean of the mold, which allow for rectal or vaginal administration to the patient and which contain at least one compound constituting the subject matter of this invention. Suppositories are one of the forms of such compositions, which can be obtained by mixing the compounds constituting the subject matter of this invention with suitable non-irritating inert environments or carriers such as cocoa butter, polyethylene glycol or wax base of the suppository, which are in a solid state at ordinary temperatures, but become liquid at body temperature and therefore melt at rectal or vaginal insertion and release the active ingredient.

The pharmaceutical composition introduced by injection, may be injected intramuscularly, intravenously, intraperitoneally and/or subcutaneously. The composition forming the subject of the present invention, is prepared in liquid solutions, in particular in physiologically compatible buffers such as solution Khanka or ringer's solution. In addition, the composition can be prepared in solid form and dissolved or suspended immediately prior to use. It is also possible lyophilized form. Preparative forms are sterile and include emulsions, su is pensii, aqueous and non-aqueous injection solutions which may contain suspendresume substances, thickeners and anti-oxidants, buffers, bacteriostats and additives that make the composition isotonic, and to have the correct pH level corresponding to the indicator of the patient's blood, which will be administered the drug.

Pharmaceutical compositions forming the subject of the present invention, suitable for nasal or inhalation use, are pharmaceutical compositions which form is suitable for administration to a patient or nasal inhalation. The composition may contain a carrier in powder form with a particle size of, for example, in the range from 1 to 500 microns (including particle sizes in the range from 20 to 500 microns in increments of 5 microns, i.e. with dimensions of 30 microns, 35 microns, etc). Suitable compositions with liquid media for use, for example, as a spray or nasal drops include aqueous or oily solutions of the active ingredient. Composition suitable for aerosol administration, can be obtained in accordance with conventional methods and to introduce other therapeutic substances. To provide inhalation therapy can be used dosing inhalers or any dry powder inhalers, such as Eclipse, Spinhaler® or Ultrahaler®, as described in the patent for which VCE WO2004/026380 and in US patent No. 5176132.

The actual dosage of the active ingredient(s) in compositions comprising the subject matter of this invention may vary with the purpose of obtaining the amount of the active ingredient(s)effective to produce a desired therapeutic effect for a particular composition and method of its introduction to the patient. Therefore, the dosage selected for each patient depends on many factors such as the desired therapeutic effect, the route of administration, the desired duration of treatment, etiology and severity of the disease, the patient's condition, weight, sex, diet, and age, type, and activity of each active ingredient, speed of absorption, metabolism and/or selection, and other factors.

Full daily dose of the compounds constituting the subject matter of the present invention, administered to the patient one or more doses can be, for example, from about 0.001 to about 100 mg/kg of body weight per day, in particular from 0.01 to 10 mg/kg/day. For example, the adult dose usually ranges from about 0.01 to about 100, in particular from about 0.01 to about 10 mg/kg of body weight per day by inhalation, from about 0.01 to about 100, in particular from 0.1 to 70, preferably from 0.5 to 10 mg/kg of body weight per day by oral administration, and from about 0.01 to about 50, in particular from 0.01 to 10 mg/kg of body weight per day intravenously. the percent concentration of active ingredient in the composition may be different, but it must be in the same proportion in order to obtain the optimal dosage. The dosage form can contain a fraction of a unit dose, allowing to obtain the desired daily dose. Obviously, possibly the almost simultaneous introduction of several standard doses. The dose may be as frequent as needed to achieve the desired therapeutic effect. Some patients are able to respond quickly to large or smaller doses, and it may be more adequate maintenance dose. For other patients may require prolonged treatment with intensity from 1 to 4 doses per day in accordance with the physiological needs of each patient. Needless to say, other patients may require no more than one or two doses per day.

Standard dose formulations may be obtained by any of the conventionally used in the pharmaceutical industry methods. Such methods include phase binding pharmaceutically active ingredient with a carrier consisting of one or more accessory ingredients. Typically, the compositions will receive a uniform and tight binding of the active ingredient with liquid carriers, or fine solid carriers, or both, with the subsequent formation of the product if necessary.

Stood the s can be packaged in a single dose or in multiple doses, for example sealed ampoules and vials with elastic tubes, and can be stored in a lyophilized condition requiring only the addition of sterile liquid carrier, for example water for injections, immediately prior to use. Individual injection solutions and suspensions may be prepared from sterile powders, granules or pellets of the above described types.

The compound forming the subject of the present invention, can be analyzed using the following analytical methods.

Liquid chromatography high pressure mass spectrometry (IHMS)

Experiments IHMS to determine retention time (RT) and associated mass ions is performed using the following method. Mass spectra (MS) recorded on a mass spectrometer Micromass LCT. The method includes the ionization positive elektrorazpredelenie and scan mass m/z 100 to 1000. Liquid chromatography carried out using a binary pump and degasser Hewlett Packard 1100 Series Binary Pump & Degasser; stationary phase: column Phenomenex Synergi 2µ Hydro-RP 20×4.0 mm, mobile phase: A=0.1% of formic acid (FA) in water, B=0,1% FA in acetonitrile. Injected volume of 5 μl using system CTC Analytical PAL. The flow rate of eluent is 1 ml/min Gradient from 10% B to 90% B over 3 minutes, and from 90% B to 100% B in 2 minutes. Supporting children who ctory: UV detector Hewlett Packard 1100 Series, wavelength = 220 nm, and evaporative light scattering detector (ELS) Sedere SEDEX 75, temperature = 46°C, the nitrogen pressure = 4 bar.

The spectra of nuclear magnetic resonance (NMR)1H

The NMR spectra at 300 MHz1H recorded at room temperature on the spectrometer Varian Mercury (300 MHz) with 5 mm ASW sensor. In the NMR chemical shift (δ) are expressed in ppm relative to tetramethylsilane. The magnitude of the chemical shift is indicated in ppm (ppm) relative to tetramethylsilane (TMS) as internal standard.

X-ray powder diffractometry (RPD)

RPD-measurement is performed on the diffractometer Siemens-Bruker D5000, using the scheme focusing on the Bragg-Brentano (theta - 2-theta). The primary acidic phosphate salt is placed on the plate of monocrystalline silicon with a surface oriented along the crystallographic plane (510). As an x-ray source using radiation from a copper target x-ray tube (45 kV/40 mA) on-line K-alpha copper (1,54056 angstroms), emission line Cu K-beta filter using a monochromator in the reflected beam. As a detector use acquired scintillation counter. The sizes of gaps establish the following: deflecting the slit of 0.6 mm, which prevents the scattering slit of 0.6 mm, the slit of the monochromator 0.1 mm receiving slit of 0.6 mm detector X-rays the recordings shall indicate the following conditions: scanning in the range of 2-40 degrees (angle 2-theta) with time accounts on the same step of 1 second, with a step of 0.02 ° at normal temperature, pressure and relative humidity.

Analysis of solvation/hydration method thermogravimetry

Thermal analysis is performed using a combined DSC/TGA analyzer (differential scanning calorimeter/thermogravimetric analyzer) TA Instruments Model Q-600 in an atmosphere of dry nitrogen. Calibration temperature TGA analyzer spend on indium standard. The primary acidic phosphate salt is placed in an aluminum cell (room 900793.901). Thermograms recorded at a linear heating rate of 10°C per minute.

Differential scanning calorimetry (DSC)

DSC analysis performed using differential scanning calorimeter (TA Instruments Model Q-1000 cooling system in a dry nitrogen atmosphere. For calibration of the DSC device using indium standard. The primary acidic phosphate salt is placed in an aluminum cell with lid fixed to ditch cold welding, the cover has a small hole formed by the laser (non cuvettes and caps 900793.901 and 900860.901 respectively). The DSC thermogram recorded at a linear heating rate of 10°C per minute.

Micrograph

Obtaining micrographs made with the help of microscope Olympus BX-41 with cross-polarization. The samples prepared through the ohms dispersion in mineral oil.

The distribution of particles sizes

The distribution of particle size measured by laser diffraction analyzer particle size Sympatec HELOS-BF with the measuring lens R3, disperser RODOS dry powders and on the laser wavelength of 632.8 nm. System calibration is performed using standard silicon carbide. The powder was dispersed by using a dispersing device for RODOS dry powder at an initial pressure of 3.0 bar and a maximum pressure drop. The distribution of particle size based on volume, calculated according to the method of the Fraunhofer using software Sympatec Windox (Version 4.0).

Dynamic sorption of water vapor (DSWP)

The profile of water sorption is determined using an analyzer dynamic sorption of water vapor, the model DVS-1 (SMS Instruments) or SGA-100 (VTI Instruments). A calibration of the relative humidity and weight comply with appropriate standards. Before the experiment the primary acidic phosphate salt is placed in the apparatus and dried at a relative humidity of ≤1% for 2.5 hours. Relative humidity speed increase from approximately 0%to 95%. It is assumed that the weight of the sample at each stage does not change, if within 5 minutes of the relative change of mass is less than 0,005% with a minimum time of establishing ravenous the I 15 minutes.

Infrared spectroscopy with Fourier transform

The FTIR-spectra were recorded using a spectrometer (Nicolet Magna-IR Spectrometer 55, paired with an IR microscope (Nicolet Nic-Plan. The investigated solid sample is placed on a KBr disk. Each spectrum recorded after 32 times of scanning in the range 4000-400 cm-1with a resolution of 4 cm-1.

The following illustrates the method of preparation and properties of compounds constituting the subject matter of this invention.

EXAMPLE

Monopotassium phosphate salt of 2-(3-{6-[2-(2,4-dichlorophenyl)ethylamino]-2-methoxypyridine-4-yl}phenyl)-2-methylpropionic acid

Stage 1. A solution of 4,6-dichloro-2-methoxypyridine (0.7 g), 2,4-dichlorophenylamino (0,82 g) and sodium hydrogen carbonate (0.88 g) in ethanol (25 ml) was heated at 80°C for three hours and was poured into 400 ml of water. The resulting solid phase was filtered and dried in the air, receiving (6-chloro-2-methoxypyridine-4-yl)-[2-(2,4-dichlorophenyl)ethyl]amine.

Stage 2. To a solution of Diisopropylamine lithium in a mixture of tetrahydrofuran/n-heptane/ethylbenzene (1.8 M, 17 ml) at 0°C dropwise within 15 minutes was added a solution of 2-(3-bromophenyl)propionic acid (3 g, a 13.9 mmol) in tetrahydrofuran (5 ml). The resulting mixture was stirred for 1 hour and then added dropwise within 10 minutes was added methyl iodide (4,93 g, 34.8 mmol) in tetrahydrofuran (5 ml). The reaction is ionic mixture was stirred for 15 hours, extinguished 2 N HCl, concentrated in vacuo and diluted with diethyl ether (150 ml). The ether layer is washed with 2 N HCl and three times was extracted with 2 N NaOH (50 ml). The combined layers of NaOH was oxidized 6 N HCl to pH 1 and extracted three times with diethyl ether (75 ml). The combined organic layers were washed with brine, dried over sodium sulfate and concentrated to obtain a solid 2-(3-bromophenyl)-2-methylpropionic acid (is 3.08 g, 91%)which was used without further purification. LC/MS: 243 (M+H).

Stage 3. To a solution of 2-(3-bromophenyl)-2-methylpropionic acid (2,18 mmol) in anhydrous ether (20 ml) was added dropwise tert-butyl lithium (a 1.7 M solution in pentane, to 5.4 ml, 9,16 mmol) at -78°C, the resulting mixture was stirred for 30 minutes and treated with tributyrate (2,34 ml 8,72 mmol). The reaction mixture was allowed to warm to room temperature, then it was stirred for 15 hours, diluted with ether and extinguished 1 M H3PO4. After stirring for 30 minutes was separated by a layer of ether and was extracted three times with 2 N NaOH (20 ml). The combined NaOH extracts were oxidized 6 N HCl to pH 1 and extracted three times with diethyl ether (50 ml). The combined organic extracts were washed with brine, dried over sodium sulfate and concentrated to obtain 3-(1-carboxy-1-methylethyl)phenylboronic acid, which was used without further purification. M IS: 209 (M+H).

Stage 4. A solution of (6-chloro-2-methoxypyridine-4-yl)-[2-(2,4-dichlorophenyl)ethyl]amine (0.51 mmol) and 3-(1-carboxy-1-methylethyl)phenylboronic acid (0.61 mmol) in acetonitrile (2.5 ml) and aqueous sodium carbonate solution (0,4 M, 2.5 ml) was degirolami by purging with nitrogen for 5 minutes, and then thereto was added tetrakis(triphenylphosphine)palladium(0) (29.5 mg, 5 mol.%). The reaction vessel was sealed and heated in the microwave to 130°C for 30 minutes. To the reaction mixture were added 2 ml of water, then using 2 N aqueous HCl solution was installed approximately pH 7, then the mixture was extracted three times with ethyl acetate (30 ml). The combined extracts were washed with brine, dried over sodium sulfate and concentrated. The residue was purified column chromatography on silica gel with getting a solid 2-(3-{6-[2-(2,4-dichlorophenyl)ethylamino]-2-methoxypyridine-4-yl}phenyl)-2-methylpropionic acid (205 mg, 75%). LC/MS: RT=2,39 min, MS: 460 (M+H);1H NMR [300 MHz, (CD3)2SO]: δ 12,38 (1H, s), of 7.36-of 8.00 (7H, m), to 6.58 (1H, s), of 3.84 (3H, s)to 3.58 (2H, m), 2,98 (2H, m), and 1.54 (6H, s).

Stage 5. Phosphoric acid (3,21 ml, 1,49 N aqueous solution) was added to a solution of 2-(3-{6-[2-(2,4-dichlorophenyl)ethylamino]-2-methoxypyridine-4-yl}phenyl)-2-methylpropionic acid (2.1 g, 4,56 mmol) in tetrahydrofuran (45 ml), the mixture was stirred for 10 minutes. Then dropwise gradually add the Lyali water until until the mixture turned into a clear solution, then continued stirring for 1.5 hours at room temperature. The mixture was concentrated in vacuo, the residue was recrystallized from acetone to obtain dihydrogenphosphate salt 2-(3-{6-[2-(2,4-dichlorophenyl)ethylamino]-2-methoxypyridine-4-yl}phenyl)-2-methylpropionic acid in powder form (2.4 g, 94%). LC/MS: RT=2,41 min; MS: 462 (M+H);1H NMR [300 MHz, (CD3SO)2SO]: δ of 7.95 (1H, usher.), 7,8 (1H, usher.), of 7.6 (2H, usher.), was 7.45 (2H, user., J=2 Hz), 7,35 (2H, s), 6,55 (1H, s), 3,85 (3H, s), 3,55 (2H, usher.), 2,95 (2H, t, J=2 Hz), 1,5 (6H, s).

Pharmacological analysis

The inhibitory effect of the compounds constituting the subject matter of the present invention, assessed using the functional analysis on the DP of the person. Applied analysis of camp using a cell line of human LS174T expressing endogenous receptor DP. A Protocol similar to that described previously (Wright D.H., Ford-Hutchinson A.W., Chadee K., Metiers C.M., The human prostanoid DP receptor stimulates mucin secretion in LS174T cells, Br. J. Pharmacol. 131(8); 1537-45 (2000)).

The SPA Protocol-analysis of camp in cells LS174 T man

Materials

- PGD2 (Cayman Chemical, cat. No. 12010)

- IBMX (Sigma cat. No. 5879)

- Direct screening SPA-analysis of camp (code Amersham RPA 559)

- 96-hole plates to cell cultures (Wallac cat. No. 1450-516)

- Scintillation counter Wallac 1450 Trilux Microplate (PerkinElmer)

- Fixtures DL the sealing tablet

- Centrifuge tubes Eppendorf

- In saline phosphate buffer (PBS) Dulbecco (Invitrogen cat. No. 14040-133)

- Distilled water

- Shaker

- Magnetic stirrer and a set of anchors

Reagent preparation:

Before dilution, all reagents should be brought to room temperature.

Analytical buffer 1X

Transfer the contents of the bottle 500-ml beaker repeated washing with distilled water. Bring total volume to 500 ml by adding distilled water, and mix thoroughly.

Lyse reagent 1 & 2

Dissolve each of the lytic reagents 1 and 2 in 200 ml of analytical buffer, respectively. Leave on for 20 minutes at room temperature to dissolve.

Granules of rabbit antibodies for SPA-analysis

Add in a bottle of 30 ml lyse buffer 2. Gently shake the bottle for 5 minutes.

Immune serum

Add 15 ml of lyse buffer 2 in each vessel and gently stir contents until dissolved.

Indicator (I125-camp)

Add 14 ml of lyse buffer 2 in each vessel and gently stir contents until dissolved.

Preparation of immunoreagent

1) Add an equal volume indicator, immune serum and SPA-reagent antibodies rabbit in the vessel, after the PE the that was prepared sufficient for the desired number of holes (150 μl per well).

2) mix Thoroughly.

3) you Must prepare a fresh solution of immunoreagent before each analysis and are not reusable.

Standard

1) Add 1 ml of lyse buffer 1 and gently stir contents until dissolved.

2) the Final solution contains camp at a concentration of 512 pmol/ml

3) Mark 7 polypropylene or polystyrene tubes "0.2 pmol", "0.4 pmol", "0.8 pmol", "1.6 pmol", "3.2 pmol", "6.4 pmol and 12.8 pmol".

4) Pipette, add 500 ál lyse buffer 1 in all test tubes.

5) In a test tube "12.8 pmol, add by pipette 500 ál of standard mixture (512 pmol/ml) and mix thoroughly. Transfer 500 ál from tube "12.8 pmol" in the tube "6.4 pmol and mix thoroughly. Repeat the dilution procedure twice with the rest of the tubes.

6) portions of 50 μl in duplicate from each serial dilution and standard source solution will allow you to get 8 standard camp levels in the range from 0.2 to 25.6 pmol.

Dilution buffer connection

Add 50 ál of 1 mm IBMX in 100 ml of PBS to obtain a final concentration of 100 μm, and process ultrasound at 30°C for 20 minutes.

Preparation PGD2

Dissolve 1 mg of PGD2 (FW, AZN 352.5) 284 μl of DMSO, which is advised to get the source solution 10 mm, and keep it at 20°C. Before each analysis it is necessary to prepare a fresh solution. Add 3 ál of 10 mm initial solution in 20 ml of DMSO, mix thoroughly and transfer 10 ml to 40 ml PBS.

Dilution connection

Several samples of the compounds constituting the subject matter of the present invention, tested in 96-well pad. Each sample connection occupies one row of 96-well plate.

The dilution of the compounds is carried out in Biomex 2000 (Beckman) using method 11 points camp DP.

5 ál of each joint of 10 mm tablet standard solutions of the compounds are transferred into wells of 96-well plates, as described below.

td align="left">
123456789101112
A1
B2
C3
D4
E5
F6
G7
HControl

Fill the tablet 45 μl of DMSO, except column 7 where add 28 μl of DMSO. Carefully measure the pipette contents of columns 1 and transfer 12 ál in parallel in column 7. Perform a serial dilution of 1:10 from column 1 to column 6 and column 7 to column 11, transferring from 5 ál 45 ál DMSO to obtain the following concentrations:

The first tabletThe final concentration
Column 120
Column 110.03 µm
Column 100,3 ám
Column 93 microns
Column 80.03 mm
Column 70.3 mm
Column 60.01 µm
Column 50.1 ám
Column 41 micron
Column 30,01 mm
Column 2Column 11 mm

Add in a new 96-well plate of 247.5 μl of dilution buffer for the connection. Transfer of 2.5 μl of serially diluted compounds from cooked above tablet in new (dilution 1:100) as follows:

The first tabletThe second tabletThe final concentration
Column 12Column 10
Column 6Column 20.1 nm
Column 11Column 30.3 nm
Column 5Column 41 nm
Column 10Column 53 nm
Column 4Column 60.01 µm
Column 9Column 70.03 µm
Column 3 Column 80.1 ám
Column 8Column 90,3 ám
Column 2Column 101 micron
Column 7Column 113 microns
Column 1Column 1210 µm

The growth of cells

1. LS174 T always grown in MEM (ATCC cat. No. 30-2003), 10% FBS (ATCC cat. No. 30-2020) and an additional 2 mm L-glutamine, at 37°C and 5% CO2.

2. Heat of 0.05% trypsin and versine (Invitrogen cat. No. 25300-054) at 37°C in a water bath.

3. Remove the environment for the growth of cells. The cells in the flask bulldozer t165 rinse twice with 4 ml of trypsin, and then incubate at 37°C and 5% CO2within 3 minutes.

4. Add 10 ml of medium and pipette carefully separate and count the cells.

5. Adjust cell density to 2.25×105cells per ml and sow 200 ál of cells per well (45,000 cells per well) in 96-well tablet 1 day before analysis.

Order analysis

DAY 1

Sow 45,000 cells per well in 200 μl of medium in 96-well plates. Incubate the tablets with the cells at 37°C, 5% CO2and 95% humidity during the night.

DAY 2

1 Follow the dilution of the connection.

2. Prepare analytical buffer lyse buffers 1 and 2, PGD2and the standard.

3. Remove the medium from the cells by aspiration and add 100 ál of a solution of the compound, following the Protocol of camp DP Zymark Sciclone-ALH/FD.

4. Incubate cells at 37°C, 5% CO2and 95% humidity for 15 minutes.

5. Add 5 ál of 300 nm PGD2 (20×15 nm final concentration) to each well using a Protocol Zymark camp DP PGD2 and incubate cells at 37°C, 5% CO2and 95% humidity for another 15 minutes.

6. Remove the medium from the cells by aspiration and add 50 ál lyse buffer 1, following the Protocol Zymark camp DP lysis, and incubate at room temperature with shaking for 30 minutes.

7. Add to the wells, 150 μl of immunoreagent (total volume 200 μl per well).

8. Seal the tablet, shake for 2 minutes and place in the camera scintillation counter for microtiter plate Wallac for 16 hours.

DAY 3

Count the number of [125I] camp for 2 minutes in a scintillation counter 1450 Trilux.

Data processing

Set aside the default dependence camp on the number of pulses per minute.

Table 1
Typical data analysis for standard
camp (pmol/ml) Pulse/minCf. pulse/min
0,2572557695530
0,4536752596317
0,8469547966507
1,6425141786581
3,2343434296601
6,4275827166711
12,8209420546680
25,6153115736653

Concentration camp (pmol/ml) of unknown samples is calculated by the standard according to camp on the number of pulses per minute. The percentage inhibition is calculated by the following formula:

Results

Monopotassium FOS is atna salt gives a 50% inhibition in the SPA-analysis of camp in T-cells LS174 person at a concentration of 0.3 nm.

Monopotassium phosphate salt has a higher solubility in water as compared to free form and hydrochloric salt. Below are the solubility of the free-form hydrochloric salt and monopotassium phosphate salts at 25°C in phosphate buffer (PBS pH 7,4, a mixture of 19% 0,1M solution of monopotassium phosphate and 81% 0,1M solution secondary acid phosphate) and the solubility of the free form and monopotassium phosphate salts at 37°C in a model of intestinal juice under conditions of starvation (“FaSSIF”, prepared by the method described in the work Dressman JB, Amidon G.L., Reppas C. and Shah V.P., Dissolution testing as a prognostic tool for oral drug absorption: immediate release dosage forms,Pharmaceutical Research:1998, Vol.15, No. 1, 11-22).

Solubility (phosphate buffer PBS pH of 7.4, 25°C, g/ml)Solubility in the environment of FaSSIF (37°C, ug/ml)
Free form<114
Hydrochloric salt5
Monopotassium phosphate salt15266

It turned out that monopotassium phosphate salt the possession is t a number of useful properties, which can find application in serial production and preparation of medicines. First, monopotassium phosphate salt has a high degree of crystallinity. In powder x-ray monopotassium phosphate salts, are presented in figure 1, the peaks are of medium intensity and resolution. No halo in the middle part of the values of the 2-theta indicates the absence or low content of amorphous phase. In addition, micrographs taken before and after grinding monopotassium phosphate salt represented on Fig show a significant reduction of particle size and consistency of the physico-chemical properties as a result of grinding.

However, sodium salt has a low degree of crystallinity. As can be seen from powder x-rays, see figure 11, obtained for sample 2 sodium salt, the peaks are broadened and have a low intensity, indicating poor crystallinity. X ray powder for sample 1 sodium salt shows that the substance is mainly in the amorphous form. In addition, the presence of halo on the x-ray patterns of both samples in the middle part of the values of the 2-theta indicates a very weak crystallinity of the substance.

In addition, it is shown that monopotassium phosphate salt exists in only one paragraph is limarino form. However, the hydrochloric salt, as it has been shown to exist in three polymorphic forms, which under certain conditions can be transmuted into one another. One of the polymorphic forms hydrochloric salt (see picture for sample 2 at Fig) subcriterion as peaks on the x-ray wide and have a low intensity. Other polymorphic forms hydrochloric salt (see picture for sample 1 at Fig) has a good crystallinity.

1. The compound of formula (III)

2. The compound according to claim 1 in crystalline form.

3. Pharmaceutical composition having the properties of an inhibitor of camp, including a pharmaceutically effective amount of a compound according to claim 1, in a mixture with a pharmaceutically acceptable carrier.



 

Same patents:

FIELD: medicine, pharmaceutics.

SUBSTANCE: invention relates to novel pyrimidine derivatives of general formula I, as well as to their diastereoisomers, enentiomers and/or pharmaceutically acceptable salts, which possess inhibiting action with respect to cyclin-dependent kinases and/or tyrosinekinases of VEGF receptor. In compound of general formula (I) Q stands for group where D, E, G, L, M and T in each case represent carbon, R1 represents hydrogen, halogen or CF3, R2 represents C1-C10-alkyl, which can optionally be disrupted with one group-C(O), C2-C10-alkinyl, C3-C10-cycloalkyl or phenyl, which is optionally substituted in one or more places in similar or different way by hydroxyl, halogen, C1-C6-alkoxy, C1-C6-alkyl, C3-C10-cycloalkyl, C1-C6-alkoxy-C1-C6-alkyl, C1-C6-alkoxy-C1-C6-alkoxy-C1-C6-alkyl or -COR8, X represents oxygen, sulphur or group -NH-, R3 represents hydroxy, halogen, CF3 or C1-C6-alkoxy, m represents 0-4, R4 represents hydrogen or group -COR8, NO2 or -SO2R7, or represents C1-C10-alkyl or C3-C10-cycloalkyl, R5 represents C1-C10-alkyl, which can be optionally substituted in one or more places, in similar or different way, by hydroxyl or C3-C10-cycloalkyl, or C3-C10-cycloalkyl, R7 represents C1-C10-alkyl, which is optionally substituted by group trimethylsilanyl (TMS), R8 represents C1-C6-alkyl, C1-C6-alkoxy. Invention also relates to intermediate compounds.

EFFECT: compounds can be applied for obtaining medication intended for treatment of cancer, selected from Kaposhi's sarcoma, Khodgkin's disease, leukemia or solid tumour, such as carcinoma of mammalian gland, kung, large intestine or prostate gland, autoimmune disease, such as psoriasis, and/or proliferative diseases, such as hemangioma or angiofibroma.

21 cl, 3 tbl, 5 ex

FIELD: chemistry.

SUBSTANCE: invention relates to novel compounds of general formula (I) , where R1 represents 3-10-member non-aromatic heterocyclic group, where group is limited by group containing nitrogen as ring-constituting atom, and nitrogen, having additional bond, or group represented by formula -NR11aR11b, where R11a and R11b can be similar or differ from each other, and each represents hydrogen, C1-6 alkyl, C3-10 cycloalkyl, C6-10 aryl, 5-10-member heteroaryl or 4-10-member non-aromatic heterocyclic group, and R11a and R11b can be substituted with substituent selected from group of substituents A or group of substituents B, and R1 can be substituted with substituent selected from group of substituents A or group of substituents B; R2 and R3 represent hydrogen; R4, R5, R6 and R7 can be similar or differ from each other, and each represents hydrogen, halogen, C1-6 alkyl; R8 represents hydrogen or C1-6 alkyl; R9 represents 3-10-member non-aromatic heterocyclic group, where group is limited by group containing nitrogen as ring-constituting atom, and nitrogen, having additional bond, or group represented by formula -NR11aR11b, where R11a and R11b have the same values as described above; n represents integer 1 or 2; and X represents group, represented by formula -C(R10)=, or nitrogen, where R10 represents hydrogen; where group of substituents A consists of halogen, hydroxyl and oxogroup; where group of constituents B consists of C1-6 alkyl, C3-10 cycloalkyl, C6-10 aryl, 5-10-member heteroaryl, 3-10-member non-aromatic heterocyclic group, C1-6 alkoxy, 5-10-member heteroaryloxy, 4-10-member non-aromatic heterocyclic oxygroup and group represented by formula -T1-T2-T3, and each group in group of substitutes B can be substituted with substituent selected from group of substituents C, where T1 represents direct bond or C1-6 alkylene, T2 represents group represented by formula -NRT1-, T3 represents hydrogen or C1-6 alkyl, and RT1 represents hydrogen or C1-6 alkyl; and where group of substutuents C consists of hydroxyl, C1-6 alkyl, 3-10-member non-aromatic heterocyclic group and di-C1-6 alkylaminogroup, to pharmaceutical composition possessing anti-tumor activity, to inhibitors of: hepatocyte growth factor receptor, angiogenesis and cancer dissemination, as well as to anti-tumor medication.

EFFECT: obtaining novel compounds which demonstrate anti-tumor activity.

31 cl, 111 ex, 18 tbl

FIELD: chemistry.

SUBSTANCE: invention relates to the new pyridine and new pyrimidine derivative, their pharmaceutically accepted salt or hydrate of the general formula (I): . The invention also relates to the pharmaceutical composition, which possesses the inhibiting activity with respect to the receptor of the growth factor of hepatocytes; to the inhibitor of the receptor of the growth factor of hepatocytes, the inhibitor of angiogenesis, the antitumor drug, the inhibitor of cancerous metastatic spreading, that contains the pharmacologically effective dose of the said compounds, its pharmaceutically acceptable salt or hydrate.

EFFECT: inhibitory activity.

27 cl, 45 tbl, 540 ex

FIELD: organic chemistry, chemical technology, medicine.

SUBSTANCE: invention relates to a method for improved synthesis of pharmacologically active compound of the formula (A): Method involves the following steps: (a) interaction of compound of the formula (I): with alkaline metal nitrite in the presence of suitable acid to yield compound of the formula (VII): (b) coupling compound of the formula (VII) with compound of the formula (VI): to yield compound of the formula (V): and (c) removal of protection from compound of the formula (V) to yield compound of the formula (A). Compound of the formula (A) possesses property of antagonist of R2T receptors, high metabolic stability and bioavailability. Also, invention relates to a novel intermediate substance of the formula (I) and methods for its synthesis, and to novel intermediate substances used in its synthesis.

EFFECT: improved method of synthesis, valuable medicinal properties of compounds.

12 cl, 4 ex

FIELD: organic chemistry, herbicides, agriculture.

SUBSTANCE: invention elates to novel derivatives of uracil of the formula [I] possessing herbicide activity, a herbicide composition based on thereof and to a method for control of weeds. In derivatives of uracil of the formula [I] the group Q-R3 represents a substituted group taken among:

wherein a heterocyclic ring can be substituted with at least a substitute of a single species taken among the group involving halogen atom, (C1-C6)-alkyl-(C1-C6)-alkoxy; Y represents oxygen, sulfur atom, imino-group or (C1-C3)-alkylimino-group; R1 represents (C1-C3)-halogenalkyl; R2 represents (C1-C3)-alkyl; R3 represents OR7, SR8 or N(R9)R10; X1 represents halogen atom, cyano-group, thiocarbamoyl or nitro-group; X2 represents hydrogen or halogen atom wherein each among R7, R8 and R10 represents independently carboxy-(C1-C6)-alkyl and other substitutes given in the invention claim; R9 represents hydrogen atom or (C1-C6)-alkyl. Also, invention relates to intermediate compounds used in preparing uracil derivatives.

EFFECT: improved preparing method, valuable properties of compounds.

40 cl, 16 sch, 12 tbl, 65 ex

The invention relates to an improved process for the preparation of compounds of formula (I), where each of the residues X and Y independently of one another denotes hydrogen, halogen, (C1-C4)-alkyl, (C1-C4)-alkoxygroup or (C1-C4)-allylthiourea, each of the last three residues is unsubstituted or substituted by one or more residues from the group comprising halogen, (C1-C4)-alkoxygroup or (C1-C4)-allylthiourea, means or di[(C1-C4)alkyl]-amino, (C3-C6-cycloalkyl, (C3-C5)-alkenyl, (C3-C5)-quinil, (C3-C5)-alkenylacyl or (C3-C5)-alkyloxy, in which the compound of formula (II) or salts thereof, where X and Y are specified in the formula (I) values are subjected to interaction with 1-6 moles of phosgene per 1 mol of the compounds of formula (II) in the presence of 2-3,5 molar equivalents of an organic aminoaniline per mole of the compounds of formula (II) and in the presence of an aprotic organic solvent at the reaction temperature in the range from -30 to +60oWith

The invention relates to new derivatives of phenylsulfonylacetate General formula (I), which are herbicide and regulating plant growth properties and can find application in agriculture

The invention relates to new substituted aminomethanesulfonic General formula (I) possessing a highly effective herbicide action, as well as the way they are received, herbicide tool based on these intermediate compounds of General formula (II)

The invention relates to an improved method for producing unsymmetrical 4,6-bis(aryloxy)pyrimidine of formula I, which are used in agriculture as pesticides, and to a new intermediate compound of formula II to obtain

The invention relates to new pyrimidine compounds or their salts with pharmaceutically acceptable acids and pharmaceutical compositions based on them

FIELD: medicine.

SUBSTANCE: invention relates to medicine, in particular, to problem of non-invasive removal of atherosclerotic deposits from main vessels. Claimed is water solution, which includes collagenase, oxypropylated β-cyclodextrin, block copolymer of ethylene- and propylene- oxide, dimethyl sulfoxide and complexone with carboxyl groups with the following component ratio, wt %: oxypropylated β-cyclodextrin - 2.0-5.5; block copolymer of ethylene oxide and propylene oxide - 0.02-2.0; dimethyl sulfoxide - 3.5-10.5; collagenase - 0.002-0.007; complexone with carboxyl groups - up to 6.5; water - the remaining part.

EFFECT: composition makes it possible to remove content of atherosclerotic deposits efficiently.

6 ex, 1 tbl

FIELD: medicine, pharmaceutics.

SUBSTANCE: invention relates to field of medicine, namely to chemical-pharmaceutical industry and deals with composition of alpha-lipoic (thioctic) acid in form of solution for infusions and to method of treating diseases selected from group, which includes alcoholic and/or diabetic polyneuropathy, coronary atherosclerosis, Botkin's disease (mild and moderate severity), liver cirrhosis, poisoning with heavy metal salts and intoxications of various etiology.

EFFECT: pharmaceutic composition possesses improved technologic properties and bioavailability, it also preserves stability in long storage.

3 cl, 1 ex, 3 tbl

FIELD: chemistry.

SUBSTANCE: invention relates to novel compounds of formula (I) or pharmaceutically acceptable salts thereof where R1 and R2 together denote a group selected form groups of formula (III-1): , where R9 denotes 1) a lower alkyl group, optionally substituted with a halogen atom or lower alkoxy group, 2) an aryl group, 3) an aralkyl group, 4) a heteroarylalkyl group, 5) a heteroaryl group, where the aryl, aralkyl, heteroarylalkyl and heteroaryl groups can be substituted with a halogen atom, lower alkyl group, optionally substituted with a lower alkoxy group or 1-3 halogen atoms, lower alkoxy group, optionally substituted with 1-3 halogen atoms, cyano group, hydroxy group, alkylsulphonyl group, cycloalkylsulphonyl group, aryl group, heteroaryl group, alkylaminocarbonyl group, alkanoyl amino group, alkyl amino group or dialkylamino group; R10 denotes a lower alkyl group, optionally substituted with 1-3 halogen atoms, or a lower alkylsulphonyl group; X9-X12 denotes a carbon atom or a nitrogen atom, where the carbon atom can be independently substituted with a lower alkyl group, optionally substituted with a halogen atom or a lower alkoxy group, lower alkoxy group, optionally substituted with a halogen atom, or a cyano group or a halogen atom; R3 denotes a) a group of formula (II-1): (ii-U where R4 and R5, taken together with a nitrogen atom, form a 5- or 6-member monocyclic ring, where the monocyclic ring may contain a substitute in form of a lower alkyl group, m1 equals 3; or b) a group of formula (II-2): , where R6 denotes a lower alkyl group or cycloalkyl group; m2 equals 1 or 2; X1-X4 all denote carbon atoms, or one of X1-X4 denotes a nitrogen atom and the rest denote carbon atoms; and where "heteroaryl" in each case relates to a 5- or 6-member aromatic ring containing 1-3 heteroatoms selected from a nitrogen atom, oxygen atom and a sulphur atom. The invention also relates to a histamine H3 receptor antagonist or inverse agonist, as well as a preventive or medicinal agent.

EFFECT: obtaining novel biologically active compounds, having histamine H3 receptor antagonist or inverse agonist activity.

11 cl, 8 ex, 1 tbl

FIELD: chemistry.

SUBSTANCE: invention relates to novel prodrugs of biologically active 2,4-pyrimidine diamine compounds with structural formula given below, hydrate, solvate and pharmaceutically acceptable salts thereof. The compounds have properties for inhibiting cascade transmission of a signal from an Fc-receptor, such as FcαRI, FcγRI, FcγRIII and FcεRI, degranulation or Syk kinase activity. Structural formula:

EFFECT: compounds can be used to treat or prevent rheumatoid arthritis and autoimmune diseases.

25 cl, 21 cl, 6 ex

FIELD: medicine.

SUBSTANCE: invention refers to medicine, namely to phytotherapy. The herbal tea contains haws, motherwort herb, hop cones, peppermint leaves, melilot herb, inula rhizomes and roots, black chokeberry fruits, centaury herb, wild strawberry leaves, melissa herb, green ginger herb, corn stigma and style, mountain ash fruits, caraway fruits in the following proportions 1:1:1:0.5:2:1:1:1:1:2:1:1:1:1 respectively.

EFFECT: use of the invention provides extended range of herbal drugs with an evident therapeutic effect for prevention and treatment of chronic heart disease.

5 ex, 3 dwg, 2 tbl

FIELD: chemistry.

SUBSTANCE: invention relates to compounds of formula in which R1 and R2 independently denote C1-6alkyl; R4 denotes phenyl, substituted with trifluoromethyl if necessary; X denotes hydrogen or methyl; and Y denotes -C(O)R, where R denotes C1-6alkyl; or Y denotes -P(O)(OR5)2, where R5 denotes hydrogen or C1-6alkyl; or pharmaceutically acceptable salts thereof. Said compounds are prodrugs of adenosine A2B receptor. The invention also relates to a pharmaceutical composition which is an adenosine A2B receptor antagonist based on the compound of formula I.

EFFECT: formula I compounds and the pharmaceutical composition can be used in treating different diseases in mammals, such as gastrointestinal disorders, immunological disorders, allergic disorders, neurological disorders, cardiovascular disorders and diseases associated with cell hyperproliferation.

13 cl, 1 tbl, 15 ex

FIELD: chemistry.

SUBSTANCE: compounds are suitable for use as kinase 1β-adrenergic receptor (βARK-1) inhibitors. The invention also relates to compositions containing such compounds and to use of compounds of formula to treat and prevent chronic heart failure, hypertension myocardial ischemia and hepatitis C viral infections (HCV) and for preventing opiate addiction. The invention also pertains to methods of producing formula (I) compounds.

EFFECT: more effective use of the compounds.

11 cl, 2 tbl

FIELD: medicine, pharmaceutics.

SUBSTANCE: compound of formula pharmaceutically acceptable salt or solvate of a compound or salt (I), ring Q represents optionally substituted monocyclic or condensed (C6-C12)aryl or optionally substituted monocyclic or condensed heteroaryl where said substitutes are chosen from: halogen; (C1-C6)alkyl optionally substituted by 1-3 halogen atoms; (C1-C6)alkylsulphonyl; phenyl optionally substituted by 1 or 2 substitutes chosen from halogen, (C1-C6)alkyl which can be substituted by 1-3 halogen atoms, groups (C1-C6)alkylamino, di(C1-C6)alkylamino, (C1-C6)alkoxy, (C1-C6)alkoxy(C1-C6)alkyl and (C1-C6)alkylthio; monocyclic or condensed heteroaryl optionally substituted by halogen; or oxo; Y1 represents a bond or -NR6-CO-, where R6 represents hydrogen, ring A represents optionally substituted a nonaromatic heterocyclyldiyl where said substitutes are chosen from (C1-C6)alkyl optionally substituted by groups hydroxy, (C1-C6)alkylamino, di(C1-C6)alkylamino, morpholino, (C1-C6)alkylaminocarbonyl, di(C1-C6)alkylaminocarbonyl; cyano; (C3-C6)cycloalkyl; (C1-C6)alkoxy; (C1-C6)alkoxy(C1-C6)alkyl; phenyl; benzyl; benzyloxymethyl; thienyl; 4-8-members monocyclic nonaromatic heterocycle having 1 or 2 heteroatoms chosen from N or O, and optionally substituted by 1 or 2 substitutes chosen from (C1-C6)alkyl, (C1-C6)alkoxy, (C1-C6)alkoxy(C1-C6)alkyl and oxo; (C1-C6)alkylamino; di(C1-C6)alkylamino; a group of formula: -Y2Z'- represents a group of formula: [Formula 2] each R7 independently represents hydrogen, (C1-C6)alkyl or (C3-C6)cycloalkyl, each of R8 and R9 independently represents hydrogen or (C1-C6)alkyl, n is equal to an integer 0 to 3, Z1 represents a bond, -O-, -S- or-NR9 - where R9 represents hydrogen, (C1-C6)alkyl, acyl or (C1-C6)alkylsulphonyl, ring B represents optionally substituted aromatic carbocyclediyl or optionally substituted aromatic heterocyclediyl where said substitutes are chosen from (C1-C6)alkyl, halogen, (C1-C6)alkoxy and oxo; Y3 represents a bond optionally substituted (C1-C6)alkylene or (C3-C6)cycloalylene, optionally interrupted -O- or optionally substituted (C2-C6)alkenylene where said substitutes are chosen from (C1-C6)alkyl, (C3-C6)cycloalkyl, halogen and (C1-C6)alkoxycarbonyl; Z2 represents COOR3; R3 represents hydrogen or (C1-C6)alkyl.

EFFECT: preparation of new compounds.

30 cl, 9 tbl, 944 ex

FIELD: medicine, pharmaceutics.

SUBSTANCE: described are new compounds of general formula (I): het-X-AB (I) where het is pentamerous N-heteroaryl, additionally containing one O, S or N atom as a heteroatom with heteroaryl with additional O atom being condensed with a benzene ring, or hexamerous N-heteroaryl; X means S; and where N-atom of N- heteroaryl residue and an X group are separated by one carbon atom; AB means 1,2,3-triazolo[4,5-d] pyrimidine-7-yl radical of general formula (II): where R3 is C1-8alkyl, phenyl, benzyl optionally substituted; R5-H, C1-8alkyl or phenyl.

EFFECT: production of new compounds for preparing a pharmaceutical composition either effective for treating cardiovascular, cancer, autoimmune diseases, stroke, neurodegenerative diseases, cystic fibrosis, or used in antithrombotic therapy.

9 cl, 11 ex, 3 tbl

FIELD: chemistry.

SUBSTANCE: invention relates to novel derivatives of isothiazole-3(2H)-OH-1,1-dioxides of formula (I) or pharmaceutically acceptable salts thereof, which can increase expression of LXR α and/or β, a pharmaceutical composition based on said derivatives, use thereof in preparing a medicinal agent, as well as novel intermediate compounds of formula (V) or salts thereof. In formulae (I), (V) R2 denotes phenyl, and R1 and R3 are as described in the claim.

EFFECT: improved properties of the derivatives.

7 cl, 9 dwg, 172 ex

FIELD: medicine, pharmaceutics.

SUBSTANCE: invention relates to chemical-pharmaceutical industry, namely to creation of medication, which possesses anti-inflammatory and regenerating action. Medication contains the following ingredients: conifer needle extract, purified oleoresin of cedar or pine, or spruce, or fir, or larch or their mixture in equal proportions, vegetable oil.

EFFECT: medication has increased effectiveness and efficiency and also extends arsenal of medications, which have anti-inflammatory and regenerating action.

7 cl, 9 ex, 2 tbl

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