RU2678455C1 - N-(3-(5-(4-chlorophenyl)-1n-pyrazolo[3,4-b]pyridine-3-carbonyl)-2,4-difluorophenyl)propane-1-sulphonamide crystalline form, active component, pharmaceutical composition and medicine - Google Patents

Abstract

FIELD: chemistry.

SUBSTANCE: invention relates to the field of organic chemistry, namely to N-(3-(5-(4-chlorophenyl)-1H-pyrazolo[3,4-b]pyridine-3-carbonyl)-2,4-difluorophenyl)propan-1-sulfonamide of formula 1 or its pharmaceutically acceptable salt. Invention also relates to a new crystalline form, an active component, a pharmaceutical composition and a drug based on compound 1.

1.

EFFECT: new compound is obtained for the prevention or treatment of proliferative diseases, characterized by BRAF kinase mutation.

10 cl, 4 dwg, 5 tbl, 10 ex

Description

The invention relates to a new inhibitor of BRAF kinase N- (3- (5- (4-chlorophenyl) -1H-pyrazolo [3,4-b] pyridin-3-carbonyl) -2,4-difluorophenyl) propan-1-sulfonamide of the formula 1, a new crystalline form, pharmaceutical composition and drug for the prevention or treatment of a proliferative disease characterized by a mutation of the BRAF kinase, preferably melanoma.

Melanoma is one of the most malignant tumors that can quickly spread through the body through the lymphogenous and hematogenous pathways. With metastasis of melanoma, the prospects for cure are very bleak. The use of chemotherapy, surgery and radiation therapy can sometimes slightly improve the quality of life of the patient, but this does not extend the life more than just a few months. The median survival for metastatic melanoma is 6–7 months. There is no standard treatment regimen.

In the presence of multiple melanoma metastases (generalization of the process), it is usually proposed to carry out regional and / or systemic chemotherapy with dacarbazine (DTIC) or carmustine (BCNU), lomustine (CCNU), cisplatin, tamoxifen or cyclophosphamide. On average, it is estimated that chemotherapy can prolong the patient’s life by only 2-3% compared with the period that he would have lived without any treatment at all.

Currently, melanoma is one of the most common types of cancer in young people aged 15 to 34 years. For patients with inoperable and metastatic melanoma, the prognosis remains disappointing - one-year survival in 25.5% of cases and five-year survival in less than 15%. About 40-60% of all cases of melanoma contain BRAF mutations at position 600, of which the most common are V600E (~ 80%) and V600K (~ 20%). They cause a constant activation of the RAF-MEK-ERK signaling pathway, which leads to stimulation of cell growth, proliferation and survival in the absence of growth factors.

The idea of targeted therapy in oncology is to create a drug that accurately hits a given target - one specific molecule, which can be responsible either for the development of cancer, or for the formation of malignant signs of cancer cells, or supporting the possibility of a malignant tumor in the patient's body. Due to the targeted drug in the cancer cell, a key molecular link is damaged - one part (molecule) with the aim that this breakdown will be significant and destroy the more complex structure - the cancer cell and, accordingly, inactivates the malignant tumor. As a result, a therapeutic effect will be achieved - rescue of a patient with cancer.

The protein blocking strategy, a product of the BRAF proto-oncogen, known as the protein kinase (serine / threonine-protein kinase BRAF), is owned by the British Richard Marais and Caroline Springer. A cell in order to activate and begin to divide must receive a special signal from the outside. Further, the signal is transmitted along the so-called signaling path - this is a cascade of biochemical reactions accompanying the transmission of an external signal from the membrane receptor on the surface of the cell membrane to the nucleus. Signal transmission can go along a short or long chain (through the activation of another cascade), be direct or indirect. The Ras / Raf / MEK / ERK cell signaling pathway is a key regulator of cell proliferation and survival. Mutations along this pathway have been identified in melanoma. Activating mutations lead to increased cell proliferation and resistance to programmed cell death. Ras protein is attached to the inner cell membrane, while Raf, MEK and ERK are cytosolic proteins. Ras mutations are observed in 10-20% of patients with melanoma and can activate Raf and P13K (phosphoinositide-3 kinase) cascades.

BRAF genes and proteins were discovered about 30 years ago. More than 13,000 publications related to the study of their molecular function, role in the regulation of biological processes, both normal and in pathologies, were published in the PubMed electronic library, which indicates the importance of studying RAF proteins. However, existing gaps in knowledge about the function and regulation of RAF proteins limit our ability to inhibit their uncontrolled activity in the treatment of deadly diseases such as cancer [Lavoie, H. and M. Therrien, Regulation of RAF protein kinases in ERK signalling . Nat. Rev. Mol. Cell. Biol, 2015.16 (5): p. 281-98.].

BRAF (B-raf or Raf kinase B) is a protein from the Raf kinase family, the main role of which is phosphorylation of mitogen kinases of activated protein kinases MEK1 / 2 and subsequent activation of ERK1 / 2. Various growth factors, inflammatory stimuli, chemokines, and other factors can activate BRAF.

BRAF is involved in various normal and pathological processes, such as cell division, actin cytoskeleton rearrangements, cell adhesion and migration. The BRAF / MEK / ERK pathway in various tissues can regulate a wide variety of processes, such as the contraction of vascular smooth muscle cells, in the nervous tissue can take part in the regulation of synapses, long-term potentiation and depression, and is also involved in the cytotoxicity of natural killers during inflammation.

A mutant constitutively active protein BRAF V600 has been found in many tumors with different frequency of occurrence. It is a promising target for targeting inhibitors and, thus, the treatment of cancer.

Identification of mutations in the BRAF gene served as the basis for the development of a new direction of targeted drugs - pathological protein inhibitors produced by the mutant gene.

In 2011, the latest targeted drug for the treatment of melanoma, Vemurafenib, developed by Plexxikon (US) and marketed under the Zelboraf brand by F. Hoffmann-La Roche Ltd, was approved for use in oncology. (Switzerland) [RU 2418800, PYRROLO [2,3-B] PYRIDINE DERIVATIVES AS PROTEINKINASE INHIBITORS, 05/20/2011].

In an international multicenter, randomized, open-label, controlled phase III study, BRIM3, the efficacy and safety of Vemurafenib compared to dacarbazine was evaluated in patients with inoperable locally advanced or metastatic melanoma with a BRAF V600 gene mutation who had not previously received treatment. The study included 675 patients. In this study, it was shown that, compared with chemotherapy, Vemurafenib reduces the risk of death by 63% (risk ratio [HR] = 0.37; p <0.0001) and, in addition, by 74% (HR = 0.26; p <0.0001) reduces the risk of worsening the course of the disease (progression-free survival is the second main goal of the study). The response to therapy (the percentage of patients whose tumor has decreased) in the group of patients receiving Vemurafenib (48.4%) was almost nine times greater than in the group of patients who received chemotherapy (5.5%; p <0, 0001). After six months, 84% of the patients from the Vemurafenib group were alive compared to 64% from the chemotherapy group. An increase in overall survival, progression-free survival, and a decrease in tumor with the use of Vemurafenib were observed in patients regardless of age, gender or risk factors for the disease. The median overall survival in the group of patients receiving Vemurafenib was 10.5 months, while the overall survival of patients receiving chemotherapy was 7.8 months.

As shown, existing and approved inhibitors (vemurafenib and debrafenib) cause the development of secondary resistance, therefore, the overwhelming number of patients ceases to respond to therapy after 6-7 months. In some cases, a combination of BRAF V600 and MEK inhibitors is prescribed.

Also, drugs of this group are not without a lot of side effects noted in clinical trials and are extremely expensive. The creation of an effective domestic drug with an action based on the inhibition of the BRAF enzyme will make it possible to increase the effectiveness of the treatment of melanoma in the Russian Federation.

Perhaps new generation BRAF V600 inhibitors with a new mechanism of action will be able to overcome the problems of secondary resistance and low inhibitor efficacy in other nosologies

The determination of the BRAF V600 mutation is of clinical importance since is a predictor of response to therapy with an inhibitor of oncogenic Raf kinase B.

Therefore, the search for new active substances that have the property of a Raf kinase B inhibitor, methods for their preparation, new drugs and prodrugs based on them, for the treatment of metastatic melanoma is an urgent and important task today.

So known compound PLX8394, which is a selective inhibitor of BRAF, is in phase I / II of clinical trials of the company Plexxikon (US) for the treatment of patients with solid tumors or fleecy leukemia cells [WO 2012109075, COMPOUNDS AND METHODS FOR KINASE MODULATION, AND INDICATIONS THEREFOR, 16.08 .2012].

Among the most well-known Raf kinase B inhibitors (including the V600E mutation) that are commercially available or under active research are various classes of chemical compounds.

Biphenyls of Bayer Pharma AG (DE) are known, for example, Sorafenib (RU 2319693, UREA DERIVATIVES (OPTIONS), PHARMACEUTICAL COMPOSITION (OPTIONS), AND METHOD FOR TREATING A DISEASE RELATED TO CANCER CANCER.

Known pyrimidinioxy-oxobenzopyranes of the companies Roche (US) and Chugai Pharmaceutical (JP), for example, RO-5126766 (RU 2428420, NEW CUMARIN DERIVATIVE HAVING ANTITUMOR ACTIVITY, 09/10/2011].

Known quinazoline derivatives of AMBIT BIOSCIENCES CORPORATION (US), for example, RXDX-105 [EA 019350, CHINAZOLINE DERIVATIVES AS RAF KINASE MODULATORS AND WAYS OF THEIR APPLICATION, 03/31/2014].

GlaxoSmithKline (US) phenylsulfonamide thiazole derivatives are known, for example, the well-known drug for the treatment of metastatic melanoma Dabrafenib [EA 019349, COMPOUNDS OF BENZOLSULFONAMIDTHIAZOL AND OXAZOLE, 03/31/2014].

Pyrazolylpyrimidine derivatives of NOVARTIS AG (CH) are known, for example, Encorafenib [EP 2470526, COMPOUNDS AND COMPOSITIONS AS PROTEIN KINASE INHIBITORS, 05.28.2014].

Known new purinilpiridinilamino-2,4-diftorfenilsulfonamid derivatives of Youai (KR), for example, UAI-201 [RU 2550038, a novel PURINILPIRIDINILAMINO-2,4-DIFTORFENILSULFONAMIDA, pharmaceutically acceptable salt thereof, process for its preparation and pharmaceutical compositions having an inhibitory activity REGARDING RAF KINASES CONTAINING THIS COMPOUND AS AN ACTIVE INGREDIENT, 05/10/2015].

Derivatives of Eternity Bioscience (US), for example, EBI-907 [WO 2014043296, AMINOISOQUINOLINE DERIVATIVES AS PROTEIN KINASE INHIBITORS, 03.20.2014] are known from pyrazolopyrimidines.

Closest to the present invention are pyrazolopyrimidines of the general formula A,

where Ar is selected from the group

L _{1 is} selected from the group —C (R ₅ R ₆ ) -, —C (O) -, —C (S) -, —N (R ₇ ) -, —O—, —S—, —S (O) -, and -S (O) ₂ -;

L _{2 is} selected from the group -N (RS) -C (O) -, -N (R ₈ ) -C (S) -, -N (RS) -S (O) -, -N (R ₈ ) -S (O) ₂ -, -N (R ₈ ) -C (O) -K (RS) -, -N (RS) -C (S) -N (R8) -, and -N (R8) -S ( O) ₂ —N (R ₈ ) - etc. [WO 2010111527, PYRAZOLO [3,4-B] PYRIDINES AS KINASE INHIBITORS AND THEIR MEDICAL USE, 09/30/2010], which are also known as inhibitors of at least one Raf kinase.

However, among the compounds of general formula A, N- (3- (5- (4-chlorophenyl) -1H-pyrazolo [3,4-b] pyridin-3-carbonyl) -2,4-difluorophenyl) propane- 1-sulfonamide. Neither its properties nor its crystalline form are known. With respect to kinase activity in the above source, the experiment was conducted with only three compounds.

Therefore, N- (3- (5- (4-chlorophenyl) -1H-pyrazolo [3,4-b] pyridin-3-carbonyl) -2,4-difluorophenyl) propan-1-sulfonamide is a new compound. It is necessary to find out whether this compound is able to crystallize and have crystalline forms. Then, if crystalline forms exist, it is necessary to find reproducible methods for the preparation of chemically stable crystalline forms and to identify new crystalline forms of this compound.

Solid stability and shelf life of active components are very important factors. A drug compound and compositions comprising it must be able to be effectively stored for significant periods of time without showing a significant change in the physicochemical properties of the active component (for example, its chemical composition, density, hygroscopicity and solubility). In addition, it is also important to present the drug in a form that is as clean as possible. In this regard, amorphous compounds can present significant problems. For example, it is more difficult to handle such compounds and include them in dosage forms compared with the crystalline compound, due to dubious solubility, and it often turns out that they are unstable and chemically contaminated. One skilled in the art will understand that if a drug can be easily obtained in a stable crystalline form, then the above problems can be solved. It is usually impossible to predict only on the basis of the molecular structure what the behavior of the compound will be during crystallization, and usually this can only be established empirically.

Thus, it is an object of the present invention to provide a novel BRAF inhibitor of N- (3- (5- (4-chlorophenyl) -1H-pyrazolo [3,4-b] pyridin-3-carbonyl) -2,4-difluorophenyl) propan-1-sulfonamide and finding its new stable crystalline forms.

The technical result of the present invention is to improve the properties of this compound when it is used, in particular, a higher dissolution rate and increased storage stability.

As a result of the work, it was found that N- (3- (5- (4-chlorophenyl) -1H-pyrazolo [3,4-b] pyridin-3-carbonyl) -2,4-difluorophenyl) propan-1-sulfonamide of the formula 1 has a pronounced activity against cell lines expressing BRAF.

This inhibitory effect was confirmed during the phosphorylation of the extracellular signal-regulating kinase and cell antiproliferation in accessible melanoma cell lines expressing the BRAF gene with V600 mutations.

The task and technical result are achieved by a new inhibitor of BRAF kinase N- (3- (5- (4-chlorophenyl) -1H-pyrazolo [3,4-b] pyridin-3-carbonyl) -2,4-difluorophenyl) propan-1 -sulfonamide of the formula 1

The subject of the present invention is a new crystalline form of the compound N- (3- (5- (4-chlorophenyl) -1H-pyrazolo [3,4-b] pyridin-3-carbonyl) -2,4-difluorophenyl) propan-1-sulfonamide formula 1, having characteristic peaks on a powder X-ray diffraction pattern at the following angles 2θ (± 0.2 °): 8.9; 13.0; 14.5; 16.4; 18.5; 19.6; 21.4; 22.3; 24.3; 25.4; 28.0.

The subject of this invention is also an active component having the property of a BRAF kinase inhibitor, which is a compound of formula 1 or a new crystalline form thereof, or a pharmaceutically acceptable salt thereof.

The subject of the invention is also a pharmaceutical composition for the prophylaxis or treatment of a proliferative disease characterized by a mutation of the BRAF kinase, containing in an effective amount an active component representing a compound of formula 1 or a new crystalline form thereof or a pharmaceutically acceptable salt thereof.

More preferred is a pharmaceutical composition for the prophylaxis or treatment of a proliferative disease characterized by a BRAF kinase mutation in which the BRAF mutation is a V600E mutation.

More preferred is a pharmaceutical composition for the prophylaxis or treatment of a proliferative disease characterized by a mutation of the BRAF kinase, wherein the proliferative disease is melanoma.

More preferred is a pharmaceutical composition for the prophylaxis or treatment of a proliferative disease characterized by a mutation of the BRAF kinase, wherein the proliferative disease is colorectal adenocarcinoma.

The pharmaceutical composition may include pharmaceutically acceptable excipients. By pharmaceutically acceptable excipients are meant diluents, excipients and / or carriers used in the pharmaceutical field. The pharmaceutical composition along with the active component of the present invention may include other active ingredients, provided that they do not cause undesirable effects, for example, allergic reactions. If it is necessary to use the pharmaceutical compositions of the present invention in clinical practice, they can be mixed for the manufacture of various forms, while they can include traditional pharmaceutical carriers; for example, oral forms (such as tablets, gelatine capsules, pills, solutions or suspensions); injection forms (such as injectable solutions or suspensions, or dry powder for injection, which only requires the addition of water for injection before use); local forms (such as ointments or solutions).

The carriers used in the pharmaceutical compositions of the present invention are carriers that are used in the pharmaceutical field to obtain common forms, including: in oral forms, binders, lubricants, disintegrants, solvents, diluents, stabilizers, suspending agents, colorless are used agents, flavoring agents of taste; antiseptic agents, solubilizers, stabilizers are used in injection forms; in local forms, bases, diluents, lubricants, antiseptic agents are used.

The pharmaceutical composition of the present invention can be obtained by mixing the active component, H- (3- (5- (4-chlorophenyl) -1H-pyrazolo [3,4-b] pyridin-3-carbonyl) -2,4-difluorophenyl) propane -1-sulfonamide, with an inert excipient, solvent and / or other pharmaceutically acceptable excipients.

The subject of the present invention is also a medicament for the prophylaxis or treatment of a proliferative disease characterized by a mutation of the BRAF kinase, in the form of tablets, capsules or injections, placed in a pharmaceutically acceptable package containing in an effective amount a compound of formula 1 or its new crystalline form, or its pharmaceutically acceptable a salt or active ingredient or a pharmaceutical composition of the present invention.

More preferred is a medicament in the form of a tablet containing, by weight. %:

N- (3- (5- (4-chlorophenyl) -1H-pyrazolo [3,4-b] pyridin-3- carbonyl) -2,4-difluorophenyl) propan-1-sulfonamide 27.6; Hypromellose acetate succinate 64.4; Silicon Colloidal Dioxide 1.15; Croscarmellose sodium 3.45; Hydroxypropyl cellulose 0.46; Magnesium stearate 0.64; Opadry White 2.30.

The subject of the present invention is a method for the prophylaxis or treatment of a proliferative disease characterized by a mutation of the BRAF kinase, according to which a needy subject is administered an effective amount of an active component having the property of a BRAF kinase inhibitor, which is a compound of formula 1 or its new crystalline form, or its pharmaceutically acceptable salt, or pharmaceutical composition or drug of the present invention.

Medicines may be administered orally or parenterally (e.g., intravenously, subcutaneously, intraperitoneally, topically or rectally). The clinical dosage of the active component (s), pharmaceutical composition or combination drug comprising a pharmaceutically effective amount of the active component in patients can be adjusted depending on the therapeutic efficacy and bioavailability of the active ingredients in the body, their metabolic rate and excretion from the body, as well as depending on the age, sex and stage of the patient’s disease, while the daily dose in adults is usually 10 ~ 1500 mg, preferably 50 ~ 1200 m g. Therefore, during the preparation of the pharmaceutical compositions of the present invention as dosage units, the above effective dosage should be taken into account, with each dosage unit of the preparation containing 10 ~ 1500 mg, preferably 50 ~ 1200 mg. In accordance with the instructions of a doctor or pharmacist, these drugs can be taken several times during certain periods of time (preferably from one to six times).

The following are definitions of terms that are used in the description of the present invention.

BRAF (Raf Kinase B), a protein of 766 amino acids, is a key element of the RAS-RAF signaling pathway that allows cell growth and existence.

“Azaheterocycle” means an aromatic or non-aromatic monocyclic or polycyclic system containing at least one nitrogen atom in a ring.

"Active component" (drug substance, drug substance, drug-substance) means a physiologically active substance of synthetic or other (biotechnological, plant, animal, microbial and other) origin, having pharmacological activity and is the active principle of the pharmaceutical composition used for the manufacture and manufacture medicinal product (means).

“Aromatic cycle” (aromatic system) means a planar cyclic system in which all atoms of the cycle participate in the formation of a single conjugation system, which, according to the Hückel rule, includes (4n + 2) π-electrons (n is a non-negative integer). Examples of aromatic cycles are benzene, naphthalene, anthracene, and the like. In the case of “heteroaromatic cycles”, π-electrons and p-electrons of heteroatoms participate in the conjugation system; their total number is also equal to (4n + 2). Examples of such cycles are pyridine, thiophene, pyrrole, furan, thiazole and the like. An aromatic ring may have one or more “ring system substituents” and may be anelated with a non-aromatic ring, heteroaromatic or heterocyclic ring.

“Heterocyclyl” means an aromatic or non-aromatic saturated monocyclic or polycyclic system comprising from 3 to 10 carbon atoms, preferably from 5 to 6 carbon atoms, in which one or more carbon atoms are replaced by a heteroatom such as nitrogen, oxygen, sulfur. The prefix "aza", "oxa" or "thia" before heterocyclyl means the presence in the cyclic system of a nitrogen atom, an oxygen atom or a sulfur atom, respectively. Heterocyclyl may have one or more “cyclic system substituents,” which may be the same or different. The nitrogen and sulfur atoms in the heterocyclyl can be oxidized to N-oxide, S-oxide or S-dioxide. Representative heterocyclyls are piperidinyl, pyrrolidinyl, piperazinyl, morpholinyl, thiomorpholinyl, thiazolidinyl, 1,4-dioxan-2-yl, tetrahydrofuranyl, tetrahydrothiophenyl, etc.

"Hydrate" means a stoichiometric or non-stoichiometric composition of a compound or its salt with water.

“Substituent” means a chemical radical that is attached to a scaffold (fragment), for example, “substituent alkyl”, “substituent of an amino group”, “substituent carbamoyl”, “substituent of the cyclic system”.

"Inhibitors" (lat. Inhibere - to delay) - the general name of substances that suppress or delay the course of physiological and physico-chemical (mainly enzymatic) processes, the study of enzyme inhibition plays an important role in creating drugs, in studying the mechanism of action and structure of enzymes.

“Medicinal product (preparation)” - a combination of several medicinal substances for simultaneous use in the form of tablets, capsules, injections, ointments, rectal suspensions and gels and other finished forms, designed to restore, correct or alter physiological functions in humans and animals, and also for the treatment and prevention of diseases, diagnosis, anesthesia, contraception, cosmetology and other things. Medicinal substances in one set can be presented in the form of various ready-made forms intended for administration into an animal or human body in various ways, for example, orally and rectally.

"Ligands" (from Latin ligo - bind) are chemicals (small molecule, inorganic ion, peptide, protein, etc.) that can interact with receptors that transform this interaction into a specific signal.

"Receptors" (from the Latin recipere - to receive, to recognize) are biological macromolecules located on the cytoplasmic membrane of a cell or intracellular, capable of specifically interacting with a limited set of physiologically active substances (ligands) and transforming the signal about this interaction into a specific cellular response.

“Signal cascade” (signal system, signal transmission cascade) means a set of interconnected sequential and parallel molecular processes of regulation of cellular metabolism by external (primary) signals that carry information into the cell, which fundamentally distinguishes them from other chemical compounds entering the cell that serve as its source matter and energy. The molecular mechanisms of transmission (transduction) of external signals into the cell imply not only the transmission of signals as such, but also the whole complex of events associated with it, including amplification, attenuation and suppression (or switching off) of signals.

"Pharmaceutical composition" means a composition comprising a compound of formula 1 and at least one of the components selected from the group consisting of pharmaceutically acceptable and pharmacologically compatible excipients, solvents, diluents, carriers, excipients, distributing and perceiving agents, agents delivery, such as preservatives, stabilizers, fillers, grinders, moisturizers, emulsifiers, suspending agents, thickeners, sweeteners, perfumes, flavors, antibacterial agents, fungicides, lubricants, and prolonged delivery controllers, the choice and suitable proportions of which depend on the nature and way of administration and dosage. Examples of suspending agents are ethoxylated isostearyl alcohol, polyoxyethylene, sorbitol and sorbitol ether, microcrystalline cellulose, aluminum metahydroxide, bentonite, agar-agar and tragacanth, as well as mixtures of these substances. Protection against the action of microorganisms can be provided with a variety of antibacterial and antifungal agents, for example, parabens, chlorobutanol, sorbic acid and the like. The composition may also include isotonic agents, for example, sugars, sodium chloride and the like. The prolonged action of the composition can be achieved using agents that slow down the absorption of the active principle, for example, aluminum monostearate and gelatin. Examples of suitable carriers, solvents, diluents and delivery vehicles are water, ethanol, polyalcohols, and also mixtures thereof, vegetable oils (such as olive oil) and injectable organic esters (such as ethyl oleate). Examples of excipients are lactose, milk sugar, sodium citrate, calcium carbonate, calcium phosphate and the like. Examples of grinders and distributors are starch, alginic acid and its salts, silicates. Examples of lubricants are magnesium stearate, sodium lauryl sulfate, talc, and high molecular weight polyethylene glycol. The pharmaceutical composition for oral, sublingual, transdermal, intramuscular, intravenous, subcutaneous, local or rectal administration of the active principle, alone or in combination with another active principle, can be administered to animals and humans in a standard administration form, in the form of a mixture with traditional pharmaceutical carriers. Suitable unit dosage forms include oral forms such as tablets, gelatine capsules, pills, powders, granules, chewing gums and oral solutions or suspensions, sublingual and buccal administration forms, aerosols, implants, local, transdermal, subcutaneous, intramuscular, intravenous, intranasal or intraocular administration forms and rectal administration forms. Pharmaceutical compositions are generally prepared using standard procedures involving the mixing of the active compound with a liquid or finely divided solid carrier. For the manufacture of suppositories, in addition to the active components, cocoa butter, its alloys with paraffin and hydrogenated fats, vegetable and animal hydrogenated fats, solid fat, lanol, alloys of hydrogenated fats with wax, hard paraffin and other bases approved for medical use are also used.

“Pharmaceutically acceptable salt” means the relatively non-toxic organic and inorganic salts of the acids and bases of the present invention. These salts can be obtained in situ during the synthesis, isolation or purification of the compounds or prepared specially. In particular, base salts can be prepared on the basis of the purified free base of the claimed compound and a suitable organic or inorganic acid. Examples of salts thus obtained are hydrochlorides, hydrobromides, sulfates, bisulfates, phosphates, nitrates, acetates, oxalates, valeriates, oleates, palmitates, stearates, laurates, borates, benzoates, lactates, tosylates, citrates, maleates, fumarates, succinates, tartrates mesylates, malonates, salicylates, propionates, ethanesulfonates, benzenesulfonates, sulfamates and the like. (A detailed description of the properties of such salts is given in Berge S. M., et al., "Pharmaceutical Salts" J. Pharm. Sci. 1977, 66: 1-19). Salts of the claimed acids can also be specially prepared by reacting the purified acid with a suitable base, and metal and amine salts can be synthesized. Metal salts include sodium, potassium, calcium, barium, zinc, magnesium, lithium and aluminum salts, the most desirable of which are sodium and potassium salts. Suitable inorganic bases from which metal salts can be obtained are hydroxide, carbonate, sodium bicarbonate and hydride, potassium hydroxide and bicarbonate, potash, lithium hydroxide, calcium hydroxide, magnesium hydroxide, zinc hydroxide. As organic bases from which salts of the claimed acids can be obtained, amines and amino acids are selected that are sufficiently basic to form a stable salt and are suitable for medical use (in particular, they should have low toxicity). Such amines include ammonia, methylamine, dimethylamine, trimethylamine, ethylamine, diethylamine, triethylamine, benzylamine, dibenzylamine, dicyclohexylamine, piperazine, ethylpiperidine, tris (hydroxymethyl) aminomethane and the like. In addition, tetraalkylammonium hydroxides, for example, such as choline, tetramethylammonium, tetraethylammonium and the like, can be used for salt formation. As amino acids, the main amino acids can be used - lysine, ornithine and arginine.

"Pharmaceutically acceptable excipients" by pharmaceutically acceptable excipients refers to pharmaceutical diluents, excipients and / or carriers.

The invention is also illustrated by the drawings.

FIG. 1. X-ray powder diffraction pattern of a new crystalline form of N- (3- (5- (4-chlorophenyl) -1H-pyrazolo [3,4-b] pyridin-3-carbonyl) -2,4-difluorophenyl) propan-1-sulfonamide formula 1 obtained according to the present invention. Dependence of intensity on angle 2θ, °.

FIG. 2. The effect of the test substances on the growth of the tumor of the A375 cell line in the xenograft model * - p <0.05 compared with the Control group).

FIG. 3. Tumor volume on day 21 of the study in the A375 xenograft model.

FIG. 4. Inhibition of tumor growth (SRW) on the 21st day of growth studies of tumor cells of the A375 line in a xenograft model (* - p <0.05 compared to the Control group).

The following examples illustrate but do not limit the invention.

Example 1. The synthesis method of the compounds of formula 1.

In a round-bottomed four-necked flask purged with nitrogen, 98.14 g of 5-bromo-3-iodo-1- (4-methoxy-benzyl) is dissolved in 981 ml of anhydrous tetrahydrofuran in a stream of nitrogen (0.5-0.6 L / min). -1H-pyrazolo [3,4-b] pyridine, the resulting solution was cooled to -40 ° C, 118 ml of 2M solution of isopropyl magnesium chloride in tetrahydrofuran were added. After 20 minutes, 58 g of N- (2,4-difluoro-3-formylphenyl) propan-1-sulfonamide are dissolved in a 736 ml of anhydrous tetrahydrofuran in a round-bottomed four-necked flask. The resulting solution was cooled to -78 ° C. 240 ml of a 1M solution of o-tolyl magnesium chloride in tetrahydrofuran was added with stirring. The resulting solutions are drained, maintaining the temperature of the reaction mixture at -40 ° C. 556 ml of 1N hydrochloric acid and 491 ml of ethyl acetate are added to the reaction mass with stirring at room temperature. The progress of the reaction is controlled by thin layer chromatography, eluent ethyl acetate: hexane 1: 2. The resulting emulsion is transferred to a separatory funnel and the organic layer is separated. The organic phase is dried over sodium sulfate and the solvent is evaporated on a rotary evaporator.

The resulting N- (3 - ((5-bromo-1- (4-methoxybenzyl) -1H-pyrazolo [3,4-b] pyridin-3-yl) (hydroxy) methyl) -2,4-difluorophenyl) propan- 1-sulfonamide is dissolved in 1164 ml of dichloromethane. While stirring in an ice cooling bath, 56.06 g of 1,1,1-tris (acetyloxy) -1,1-dihydro-1,2-benzodiodioxol-3- (1H) -one is added portionwise while maintaining the temperature at 20 ° C. The ice bath is removed and the reaction mass is stirred at 20 ° C for 2 hours. The resulting product was mixed with dioxane (1052 ml), water (349 ml), 4-chlorophenyl boronic acid (14.96 g) and potassium carbonate (52.88 g). The reaction mass is kept under vacuum for 30 minutes, then purged with nitrogen (0.5-0.6 l / min), bringing the pressure to atmospheric. Then make tetrakis (triphenylphosphine) palladium (5.53 g). The resulting mixture is again vacuum (with stirring for 5 minutes, then purged with nitrogen (0.5-0.6 l / min), bringing the pressure to atmospheric, then heated to 80 ° C. The reaction mass is kept at a temperature of 80 ° C for 16 hours, then filtered through zeolite.

The resulting N- (3- (5- (4-chlorophenyl) -1- (4-methoxybenzyl) -1H-pyrazolo [3,4-b] pyridin-3-carbonyl) -2,4-difluorophenyl) propan- 1-sulfonamide is washed with hexane and dissolved in 388 ml of concentrated sulfuric acid, maintaining a temperature of 20 ° C. The reaction mass is poured into 2717 g of ice with stirring and neutralized by adding 1085 g of potassium carbonate in small portions. Then add 388 ml of ethyl acetate. The precipitated inorganic salt precipitate was filtered off and washed with 388 ml of ethyl acetate. The emulsion is transferred to a separatory funnel, the organic phase is separated off and dried over sodium sulfate and the solvent is evaporated on a rotary evaporator. 37.69 g of N- (3- (5- (4-chlorophenyl) -1H-pyrazolo [3,4-b] pyridin-3-carbonyl) -2,4-difluorophenyl) propan-1-sulfonamide of the formula 1 are obtained. Yield 94.0%. LCMS m / z 492 (M + 1) ⁺ .

To obtain a pharmaceutically acceptable salt to the N- (3- (5- (4-chlorophenyl) -1H-pyrazolo [3,4-b] pyridin-3-carbonyl) -2,4-difluorophenyl) propan-1-sulfonamide of the formula 1 in methanol (30 ml), a 1.25 M solution of HCl in methanol or solutions of other appropriate acids are added according to known methods.

Example 2. Obtaining a crystalline form of the compounds of formula 1.

10 mg of N- (3- (5- (4-chlorophenyl) -1H-pyrazolo [3,4-b] pyridin-3-carbonyl) -2,4-difluorophenyl) propan-1-sulfonamide of the formula 1 are dissolved in 10 ml methanol, heating the mixture to 42 ° C. To the solution obtained as a result, 10 ml of water was added dropwise at the same temperature over 20 minutes, and the resulting mixture was cooled to 5 ° C during the day, after which it was filtered off and the crystalline substance was collected. The crystals are dried at 50 ° C. under reduced pressure until the crystal mass ceases to decrease upon further drying, resulting in 9.52 mg of a white solid.

X-ray powder analysis of the obtained crystalline form of N- (3- (5- (4-chlorophenyl) -1H-pyrazolo [3,4-b] pyridin-3-carbonyl) -2,4-difluorophenyl) propan-1-sulfonamide of the formula 1 carried out using a Siemens D500 diffractometer (copper radiation CuKα, graphite monochromator on a secondary beam). The full-profile diffractogram was measured in the range of angles 2 <2θ <60 ° with a step of 0.01 °. Lanthanum hexaboride was used as an external standard.

On the spectrum of a powder X-ray diffractogram of a crystalline substance (Fig. 1), characteristic peaks are observed at diffraction angles of 2θ (± 0.2 °): 8.9; 13.0; 14.5; 16.4; 18.5; 19.6; 21.4; 22.3; 24.3; 25.4; 28.0.

This crystalline form does not form solvates with organic solvents, mp = 222 ° C, practically insoluble in water.

Example 3. Determination of the specific antitumor activity of the compounds of formula 1 in vitro.

The cell lines expressing BRAF were selected as a model object on which the study of the specific antitumor activity of the compound of formula 1 was to be conducted:

A375 - human melanoma cells;

HCT116 - human colorectal carcinoma cells;

HT29 - human colorectal adenocarcinoma cells.

Protocols for culturing, subculturing and planting cells are performed in accordance with the instructions for cell lines. Cells are counted using an automatic cell counter, and the concentration of A375 cells is adjusted to 1.6 * 10 ⁴ cells / ml (3000 cells per well), the concentration of HCT116 cells is adjusted to 1.1 * 10 ⁴ cells / ml (2000 cells per well), the concentration of HT29 cells adjusted to 2.7 * 10 ⁴ cells / ml (5000 cells per well)

A tubercidin preparation was chosen as a control compound in a study of the specific antitumor activity of a compound of formula 1 in vitro. This is due to the fact that compounds that cause 100% cell death, regardless of the presence or absence of certain mutations, are used to test the quality of an essay in vitro. This allows you to more accurately build dose-dependent curves and calculate the value of the IC ₅₀ .

The cytotoxicity of these substances was evaluated using a standard MTT assay with (3- (4,5-dimethylthiazol-2-yl) -2,5-diphenyl) tetrazolium bromide.

The test compound and the control substance tubercidin are dissolved in DMSO at a concentration of 10 mM. Prepare a test compound dilution in DMSO with step 3 (threefold serial dilution) using Biomek 2000, starting at 10 mM, and the control substance tubercidin, starting at 1 mM. The minimum volume in each well is 100 μl. Using a multichannel dispenser, test compound and control substance tubercidin are added to 180 μl of cells from an intermediate dilution of 20 μl to each well in triplicate. 20 μl of medium containing DMSO instead of the test compound is added to control wells.

It is centrifuged at 500 rpm, incubated at 37 ° C, 5% CO ₂ for 5 days, after which 10 μl of a solution of MTT in PBS is added to a concentration of 5 mg / μl. MTT reagent plates are processed on a CLARIOstar plate reader, measuring absorbance at a wavelength of 570 nm for each well. Reagent MTT allows you to evaluate the overall viability of cells relative to control conditions.

An IC ₅₀ value was used as a quantitative parameter for assessing cytotoxicity, which corresponds to the concentration of a substance at which cell viability is 50%. IC ₅₀ values are calculated using GraphPadPrism 6 (GraphPadSoftware, Inc.) based on raw data.

According to the test results, the test compound of formula 1 actively inhibits the proliferation of cells of the A375 line (human melanoma), IC ₅₀ for two batches of the drug are presented in table 1.

The test compound of formula 1 actively inhibits the proliferation of HT29 cell line (human colorectal adenocarcinoma), IC ₅₀ for two batches of the drug are presented in table 1.

The test compound of formula 1 is inactive with respect to HCT116 cells (human colorectal carcinoma), and IC _{50 was} not established during the experiment.

According to the results of the studies, it was found that the compound of formula 1 exhibits a strong cytotoxic effect in relation to the A375 cell line and less pronounced in relation to the HT29 line. The HCT116 cell line is resistant to the compounds of formula 1.

Example 4. Determination of the specific antitumor activity of the compounds of formula 1 in vivo.

The study of the antitumor activity of the compound of formula 1 is carried out on a xenograft model of human melanoma line A375 in comparison with the drug Vemurafenib.

The study includes:

1) Planting immunodeficient male SCID mice at the age of 13 weeks of human melanoma cells of the A375 line,

2) the study of the effect of the compounds of formula 1 on the mass of animals,

3) studying the effect of the compounds of formula 1 on tumor growth in animals,

4) Calculation of SRW (inhibition of tumor growth).

Immunodeficient mice are a convenient model for studying the antitumor activity of drugs. The lack of immunity allows them to transplant human cancer cells, which increases the prognostic reliability of the activity of the studied substances.

Animals are kept under controlled environmental conditions (22-26 ° C and 30-70% relative humidity) in individually ventilated cages. In the rooms for keeping animals, a 12-hour lighting cycle and a 10-15-fold change in air volume per hour are supported. Animals have free access to water and feed.

Human A375 melanoma cells were grown in vials in DMEM supplemented with 10% FBS in a humid atmosphere at 95% air and 5% CO ₂ at 37 ° C. As they reached 100% confluency, the cells were reseeded: on average once every 3-4 days. With detachment, the culture fluid was removed, washed in phosphate-buffered saline and a 0.25% trypsin-EDTA solution with Hanks salts was added. Incubated for 5 minutes at 37 ° C.

At the end of the experiment, all animals were euthanized by CO ₂ inhalation.

Before implantation of the cells, the corresponding area of the mouse skin was shaved and treated with AHD-2000 disinfectant solution. After implantation of the cells, the measurement of tumors was performed every 2 days. When the average tumor size reached 100-200 mm ³ , the animals were divided into experimental groups and treatment with the studied drugs was started. The drugs were administered intragastrically, daily 2 times a day, for the 21st day. The control group received a solvent. The distribution of animals into groups is shown in Table 2.

Tumor volume was determined by the formula:

where L - corresponds to the largest diameter of the tumor, W - to the smallest. Measurements were carried out using a caliper.

The antitumor activity of the studied drugs was determined by the inhibition of tumor growth relative to tumors in the control group receiving the solvent. For this, the values of inhibition of tumor growth were calculated:

where C is the average tumor size in the control group treated with the solvent,

T is the size of the tumor in the experimental group.

Repeated measures ANOVA were used to statistically compare repeating measurements — graphs of animal body mass and changes in tumor volume during the experiment. In the case of differences between the groups, Newman-Cales post-analysis was used. Differences are determined at a statistical significance level p <0.05.

According to the results of a clinical examination throughout the experiment, the administration of the test substances does not affect the change in body weight of animals

In FIG. Figure 2 shows that in animals in the Control group, the tumor size is uniformly and constantly increasing. Daily administration of the compounds of formula 1 at doses of 50, 100 and 200 mg / kg 2 times a day dose-dependently reduced the growth rate of tumors, starting from the 17th day. At the same time, Vemurafenib at a dose of 200 mg / kg and the compound of formula 1 at a dose of 25 mg / kg also did not significantly affect the growth of tumors.

When comparing tumor sizes and inhibition of tumor growth (TPO) on the 21st day of the study (Fig. 3 and 4), it was shown that daily administration of a compound of formula 1 at a dose of 200 mg / kg leads to a significant decrease in tumor size, the TPO value was 77, 5%.

The results of the study show that daily intragastric administration of a compound of formula 1 at a dose of 200 mg / kg 2 times a day (400 mg / kg / day) leads to a statistically significant inhibition of tumor growth of the A375 cell line in a xenograft model.

The use of the compounds of formula 1 in a total dose of 400 mg / kg / day led to a statistically significant decrease in tumor volume on the 21st day of the study compared with the control group. The parameter TPO (inhibition of tumor growth) was about 80%.

Based on the results, it can be concluded that the effective single dose of the compound of formula 1 is 200 mg / kg, calculated on a per person basis.

Example 5. Determination of acute toxicity of the compounds of formula 1.

The aim of the study of acute toxicity is to determine the tolerable toxic and lethal doses of the compounds of formula 1 with a single intragastric and intraperitoneal administration to male and female mice.

The preclinical study of acute toxicity of the compounds of formula 1 was carried out according to standard methods in accordance with the Methodological guidelines set forth in the Guide to conducting preclinical studies of drugs, M., Grif and K, 2012 on outbred mice. In total, 96 animals (48 males and 48 females) participated in the experiment. The mass of animals varied depending on gender (20-22 g for males and 19-21 g for females).

The routes of administration were used:

- Intragastric (route of administration similar to the planned clinical);

- Intraperitoneal (as a method similar to intravenous administration).

In the study of acute toxicity, the doses of the test substance were selected in such a way as to determine the dose that does not cause the death of animals; a dose causing a 100% mortality rate and a dose causing a mortality rate not reaching 100%. For the study, the following doses of the compound of formula 1 were selected:

For intragastric administration - 2000 mg / kg, 1000 mg / kg and 500 mg / kg

For intraperitoneal administration - 750 mg / kg, 500 mg / kg and 250 mg / kg

Solutions of the compounds of formula 1 were prepared in a 0.1% aqueous solution of Tween-80. The studied compounds of formula 1 are administered intragastrically using a probe in a volume of 10 ml / kg, intravenous administration is carried out through a syringe into the abdominal cavity.

Based on the data obtained, LD ₅₀ , LD ₁₀ , LD ₈₄ and LD ₁₀₀ were calculated using probit analysis (Finney method, Biostat 2009 software).

According to the probit analysis, the LD _{50 of the} compound of formula 1 for male mice with intragastric administration is 975.5 ± 280.5 mg / kg; for females - 1095.1 ± 267.2 mg / kg; for male mice with intraperitoneal administration of 462.3 ± 83.3 mg / kg; for females - 425.1 ± 75.1 mg / kg.

Example 6. Determination of the kinetics of dissolution of the crystalline form of the compounds of formula 1.

The dissolution kinetics of a new crystalline form of N- (3- (5 - (4-chlorophenyl) -1H-pyrazolo [3,4-b] pyridin-3-carbonyl) -2,4-difluorophenyl) propan-1-sulfonamide of the formula 1, obtained in Example 2, is compared with the kinetics of dissolution of the amorphous compounds of formula 1 obtained in Example 1. The device for determining the dissolution rate is a three-necked vessel with a capacity of 1 liter A thermometer is introduced into one of the tubes, a glass tube for sampling and their integration in the other, and the main part of the device is a cylindrical basket 3.6 cm high and 2.5 cm in diameter, made of stainless steel in the form of a mesh with holes 40 mesh diameter (about 0.351 mm). The basket is mounted on the axis of the motor.

A dissolving medium (1000 ml) is poured into the vessel, in this experiment DMSO, since it is an important bipolar aprotic solvent and, due to its strong dissolving ability, DMSO is often used as a solvent in chemical reactions and a biological screening system. The test sample is placed in a cylindrical basket, which is installed at a distance of 2 cm from the bottom of the vessel.

For comparative sample 1, 200 mg of the new crystalline form of N- (3- (5- (4-chlorophenyl) -1H-pyrazolo [3,4-b] pyridin-3-carbonyl) -2,4-difluorophenyl) propan-1 is used -sulfonamide of the formula 1 obtained in Example 2, and for a comparative sample 2-200 mg of an amorphous compound of the formula 1 obtained in Example 1, to obtain, after complete dissolution, a solution containing 200 ppm connections.

The temperature of the solvent medium during the experiment is kept constant (37 ± 0.5 ° C). The rotation speed of the basket in the medium is regulated with an accuracy of ± 5%, it is 200 rpm. At set intervals, 1-2 ml samples are taken for analysis to determine the drug content. The taken volume of solvent is immediately replaced with a new one.

The results are shown in table 3 (where the values are the number (ppm) of samples 1 or 2 in solution).

The results demonstrate that the dissolution rate of the new crystalline form of N- (3- (5- (4-chlorophenyl) -1H-pyrazolo [3,4-b] pyridin-3-carbonyl) -2,4-difluorophenyl) propan-1- the sulfonamide of formula 1 obtained in Example 2 in DMSO is higher than the dissolution rate of the amorphous compound of formula 1 obtained in Example 1. In particular, the time during which 50% dissolution of the new crystalline form of N- (3- (5- (4 -chlorophenyl) -1H-pyrazolo [3,4-b] pyridin-3-carbonyl) -2,4-difluorophenyl) propane-1-sulfonamide of formula 1, two times less than for a comparative sample of amorph th compound of formula 1.

Example 7. Investigation of the storage of crystalline form.

All samples were stored in glass vials, sealed with rubber stoppers with aluminum caps, in a refrigerator at a temperature of ~ 4.7 ° C and in a freezer at a temperature of about -23 ° C. The amount of impurities was determined by the internal normalization method; detection was carried out by a UV detector at a wavelength of 220 nm.

Table 4 presents the test results of a new crystalline form of N- (3- (5- (4-chlorophenyl) -1H-pyrazolo [3,4-b] pyridin-3-carbonyl) -2,4-difluorophenyl) propan-1- sulfonamide of the formula 1 obtained in Example 2 (Sample 1), indicating an improvement in its stability at a reduced temperature compared to the amorphous compound of the formula 1 obtained in Example 1 (Sample 2).

The results clearly indicate the promise of the proposed new crystalline form of N- (3- (5- (4-chlorophenyl) -1H-pyrazolo [3,4-b] pyridin-3-carbonyl) -2,4-difluorophenyl) propan-1- sulfonamide of the formula 1.

Example 8. Obtaining the finished dosage form of the compounds of formula 1.

480.0 g of the compound of formula 1, 1120.0 g of hypromellose acetate succinate, 20.0 g of colloidal silicon dioxide, 60.0 g of croscarmellose sodium, 8.0 g of hyprolysis (hydroxypropyl cellulose), 12.0 g of magnesium stearate are sieved in a laboratory sieve with a nominal hole diameter of 0.5 mm, load the prepared raw materials into the working chamber of the mixer-granulator and mix.

A humidifier in the amount of 500 ml is fed through the nozzle into the working chamber (purified water - 313.0 g, alcohol 96% - 187.0 g). The resulting wet mass is discharged using a spatula and dried in a granulator dryer.

In the STD-12 mixing unit, carefully, avoiding dust formation, load the obtained granulate, and mix for 20 minutes at a stirring speed of 10 ±) rpm.

The resulting tabletting mass is discharged into a technological container. The content of the compounds of formula 1 in terms of the average weight of one tablet should be in the range from 228.0 mg to 252.0 mg.

The resulting mass is tabletted using a TP-13 tablet press with a dust collector and a tablet reject device. At the end of tabletting, the cores are film-coated.

To obtain a film shell with the mixer turned on at a speed of 400 ± 5 rpm, the following materials are loaded into the container:

- purified water - 300.0 g;

- Opadry White - 40.0 g.

The resulting mass is stirred for 45 ± 5 minutes, the resulting suspension is filtered through a sieve with a nominal hole diameter of 0.25 mm. The filtered suspension should be homogeneous, without lumps.

1640.0 g tablet cores are loaded into a film coating machine, heated for 5 minutes at an inlet air temperature of 60 ± 2 ° C and a drum rotation speed of 6 ± 1 rpm.

Using a peristaltic pump, the film-forming suspension is started to be fed in an amount of 354.0 g. The coating process for tablets - cores is carried out until tablets weighing 870.0 ± 42.0 mg are obtained, after which they are dried for 10 minutes and cooled.

Tablets are packaged in 8 tablets per blister pack of polyvinyl chloride film and varnished aluminum foil printed on a BM-17 blister machine and 7 blisters are packaged together with the instructions for use and placed in a pack of cardboard.

Example 9. Obtaining a drug in the form of capsules. Thoroughly mix the compound of formula 1 with lactose powder in a ratio of 2: 1. The resulting powder mixture is packaged in 300 mg in a suitable size gelatin capsule.

Example 10. Obtaining a medicinal product in the form of injection compositions for intramuscular, intraperitoneal or subcutaneous injection. 500 mg of the compound of formula 1 are mixed with 300 mg of chlorobutanol, 2 ml of propylene glycol and 100 ml of injection water. The resulting solution is filtered and placed in 1 ml ampoules, which are sealed.

The present invention can be used in medicine, veterinary medicine, biochemistry.

Claims (12)

1. The compound N- (3- (5- (4-chlorophenyl) -1H-pyrazolo [3,4-b] pyridin-3-carbonyl) -2,4-difluorophenyl) propan-1-sulfonamide of the formula 1, or a pharmaceutically acceptable salt

2. The crystalline form of the compound N- (3- (5- (4-chlorophenyl) -1H-pyrazolo [3,4-b] pyridin-3-carbonyl) -2,4-difluorophenyl) propan-1-sulfonamide of the formula 1, having characteristic peaks on a powder X-ray diffractogram at the following angles 2θ (± 0.2 °): 8.9; 13.0; 14.5; 16.4; 18.5; 19.6; 21.4; 22.3; 24.3; 25.4; 28.0. 3. An active component having the property of a BRAF kinase inhibitor, which is a compound of formula 1 according to claim 1 or a crystalline form according to claim 2. 4. A pharmaceutical composition for the prophylaxis or treatment of a proliferative disease characterized by a mutation of the BRAF kinase, containing in an effective amount the active component according to claim 3. 5. The pharmaceutical composition according to claim 4, characterized in that the BRAF mutation is a V600E mutation. 6. The pharmaceutical composition according to claim 4, characterized in that the proliferative disease is melanoma. 7. The pharmaceutical composition according to claim 4, characterized in that the proliferative disease is a colorectal adenocarcinoma. 8. A medicament for the prophylaxis or treatment of a proliferative disease characterized by a mutation of the BRAF kinase in the form of tablets, capsules or injections in a pharmaceutically acceptable package containing in an effective amount a compound of formula 1, or a pharmaceutically acceptable salt thereof, according to claim 1 or crystalline a form according to claim 2, or an active component according to claim 3, or a pharmaceutical composition according to any one of claims. 4-7. 9. The drug according to claim 8 in the form of a tablet containing, wt.%: N- (3- (5- (4-chlorophenyl) -1H-pyrazolo [3,4-b] pyridin-3- carbonyl) -2,4-difluorophenyl) propan-1-sulfonamide 27.6 Hypromellose acetate succinate 64,4 Silicon Colloidal Dioxide 1.15 Croscarmellose sodium 3.45 Hydroxypropyl cellulose 0.46 Magnesium stearate 0.64 Opadry White 2,30 10. A method for the prevention or treatment of a proliferative disease characterized by a mutation of the BRAF kinase, according to which a subject in need is administered a compound of formula 1, or a pharmaceutically acceptable salt thereof, according to claim 1 or a crystalline form according to claim 2, or an active component according to claim 3, or the pharmaceutical composition according to any one of paragraphs. 4-7 or a drug according to any one of paragraphs. 8 and 9.

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